SEQUENCE LISTING
The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled BIOL0369USEQ_ST25.txt, created on Dec. 8, 2021, which is 1 MB in size. The information in the electronic format of the sequence listing is incorporated herein by reference in its entirety.
FIELD
Provided herein are compounds, methods, and pharmaceutical compositions for reducing an amount of Factor XII (FXII) RNA in a cell or a subject, and in certain instances reducing the amount of FXII protein in a cell or a subject. Such compounds, methods, and pharmaceutical compositions are useful to treat or prevent a thromboembolic condition. Such thromboembolic conditions include myocardial infarction (MI), stroke, limb ischemia and necrosis, and venous thromboembolism (VTE), including deep vein thrombosis (DVT) and pulmonary embolism (PE).
BACKGROUND
The human F12 gene encodes FXII protein, a zymogen that circulates in the blood stream. The zymogen is converted to an active enzyme (FXIIa) by plasma kallikrein (PK) or by its unique ability to auto-activate following binding to artificial or biologic surfaces. In vivo, FXIIa triggers the kallikrein-kinin system leading to release of bradykinin (BK) which induces microvascular permeability, nitric oxide-mediated vasodilation, hypotension, and inflammatory reactions such as swelling, hyperperfusion, and pain. FXIIa also initiates coagulation via the intrinsic pathway and promotes inflammation via the kallikrein-kinin system comprising, high molecular weight kininogen (HK) and PK.
FXII may contribute to a variety of inflammatory, life-threatening disease conditions, including hereditary angioedema (HAE) and thrombosis. HAE is a rare life-threatening inherited disorder characterized by recurrent episodes of acute swelling in multiple organs because of increased vascular permeability. An excess of FXII activity may trigger excessive BK formation or signaling, thereby contributing to the vascular permeability of hereditary angioedema. Thrombosis is a maladaptive process of vascular occlusion and remains a primary cause of vascular morbidity and mortality associated with myocardial infarction (MI), stroke, limb ischemia and necrosis, and venous thromboembolism (VTE), including deep vein thrombosis (DVT) and pulmonary embolism (PE). VTE is the third leading cause of vascular-related death, after MI and stroke, and is one of the leading causes of death in patients with cancer. While many thromboembolic conditions are due to acquired extrinsic issues, (e.g. surgery, cancer, immobility), other thromboembolic conditions are due to a genetic mutation, (e.g., antiphospholipid syndrome, Factor V Leiden thrombophilia). The most commonly used treatments for thromboembolic conditions (e.g., warfarin, heparin) and newer direct oral anticoagulants all possess significant drawbacks, such as an increased risk of bleeding. It is therefore an object herein to provide compounds, methods, and pharmaceutical compositions for the treatment and prevention of such thromboembolic conditions.
SUMMARY OF THE INVENTION
Provided herein are compounds, methods and pharmaceutical compositions for reducing an amount of FXII RNA, and in certain embodiments reducing the amount or activity of FXII protein in a cell or a subject. In certain embodiments, compounds, methods and pharmaceutical compositions disclosed herein reduce FXII RNA, FXII protein, FXII activity, or a combination thereof, in the blood of a subject. In certain embodiments, the subject has or is at risk for a thromboembolic condition. In certain embodiments, the thromboembolic condition is deep vein thrombosis, venous thrombosis, arterial thrombosis, pulmonary embolism, myocardial infarction, or stroke. In certain embodiments, compounds useful for reducing the amount of FXII RNA, the amount of FXII protein, or FXII activity are oligomeric compounds. In certain embodiments, oligomeric compounds comprise modified oligonucleotides.
Also provided are herein are methods useful for preventing a thromboembolic condition and methods useful for ameliorating a symptom of a thromboembolic condition. Exemplary symptoms of a thromboembolic condition include, but are not limited to, pain, shortness of breath, heart burn, cold sweat, fatigue, lightheadedness, dizziness, swelling, cramping, and death.
Also provided are herein are methods useful for treating hereditary angioedema and methods useful for ameliorating a symptom of hereditary angioedema. Exemplary symptoms of hereditary angioedema include, but are not limited to, swelling, nausea, vomiting, itching, headache, fatigue, abdominal pain, shortness of breath, rhinitis, anaphylaxis, and bronchoconstriction.
DETAILED DESCRIPTION OF THE INVENTION
It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive. Herein, the use of the singular includes the plural unless specifically stated otherwise. As used herein, the use of “or” means “and/or” unless stated otherwise. Furthermore, the use of the term “including” as well as other forms, such as “includes” and “included”, is not limiting. Also, terms such as “element” or “component” encompass both elements and components comprising one unit and elements and components that comprise more than one subunit, unless specifically stated otherwise.
The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents, or portions of documents, cited in this application, including, but not limited to, patents, patent applications, articles, books, and treatises, are hereby expressly incorporated-by-reference for the portions of the document discussed herein, as well as in their entirety.
Definitions
Unless specific definitions are provided, the nomenclature used in connection with, and the procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well-known and commonly used in the art. Where permitted, all patents, applications, published applications and other publications and other data referred to throughout in the disclosure are incorporated by reference herein in their entirety.
Unless otherwise indicated, the following terms have the following meanings:
Definitions
As used herein, “2′-deoxynucleoside” means a nucleoside comprising a 2′-H(H) deoxyribosyl sugar moiety. In certain embodiments, a 2′-deoxynucleoside is a 2′-β-D-deoxynucleoside and comprises a 2′-β-D-deoxyribosyl sugar moiety, which has the β-D configuration as found in naturally occurring deoxyribonucleic acids (DNA). In certain embodiments, a 2′-deoxynucleoside may comprise a modified nucleobase or may comprise an RNA nucleobase (uracil).
As used herein, “2′-MOE” or “2′-MOE sugar moiety” means a 2′-OCH2CH2OCH3 group in place of the 2′-OH group of a ribosyl sugar moiety. “MOE” means methoxyethyl. Unless otherwise indicated, a 2′-MOE has the β-D stereochemical configuration.
As used herein, “2′-MOE nucleoside” means a nucleoside comprising a 2′-MOE sugar moiety.
As used herein, “2′-OMe” or “2′-O-methyl sugar moiety” means a 2′-OCH3 group in place of the 2′-OH group of a ribosyl sugar moiety. Unless otherwise indicated, a 2′-OMe has the β-D stereochemical configuration.
As used herein, “2′-OMe nucleoside” means a nucleoside comprising a 2′-OMe sugar moiety.
As used herein, “2′-substituted nucleoside” means a nucleoside comprising a 2′-substituted sugar moiety. As used herein, “2′-substituted” in reference to a sugar moiety means a sugar moiety comprising at least one 2′-substituent group other than H or OH.
As used herein, “5-methyl cytosine” means a cytosine modified with a methyl group attached to the 5 position. A 5-methyl cytosine is a modified nucleobase.
As used herein, “administering” means providing a pharmaceutical agent to a subject.
As used herein, “ameliorate” in reference to a treatment means improvement in at least one symptom relative to the same symptom in the absence of the treatment. In certain embodiments, amelioration is the reduction in the severity or frequency of a symptom or the delayed onset or slowing of progression in the severity or frequency of a symptom.
As used herein, “antisense activity” means any detectable and/or measurable change attributable to the hybridization of an antisense compound to its target nucleic acid. In certain embodiments, antisense activity is a decrease in the amount or expression of a target nucleic acid or protein encoded by such target nucleic acid compared to target nucleic acid levels or target protein levels in the absence of the antisense compound.
As used herein, “antisense compound” means an oligomeric compound capable of achieving at least one antisense activity.
As used herein, “bicyclic nucleoside” or “BNA” means a nucleoside comprising a bicyclic sugar moiety.
As used herein, “bicyclic sugar” or “bicyclic sugar moiety” means a modified sugar moiety comprising two rings, wherein the second ring is formed via a bridge connecting two of the atoms of the first ring thereby forming a bicyclic structure. In certain embodiments, the first ring of the bicyclic sugar moiety is a furanosyl moiety. In certain embodiments, the bicyclic sugar moiety does not comprise a furanosyl moiety.
As used herein, “cell-targeting moiety” means a conjugate moiety that interacts with a cell or a portion thereof. In certain embodiments, the cell-targeting moiety binds a receptor on a surface of a cell.
As used herein, “cEt nucleoside” means a nucleoside comprising a cEt modified sugar moiety.
As used herein, “chirally enriched population” means a plurality of molecules of identical molecular formula, wherein the number or percentage of molecules within the population that contain a particular stereochemical configuration at a particular chiral center is greater than the number or percentage of molecules expected to contain the same particular stereochemical configuration at the same particular chiral center within the population if the particular chiral center were stereorandom. Chirally enriched populations of molecules having multiple chiral centers within each molecule may contain one or more stereorandom chiral centers. In certain embodiments, the molecules are modified oligonucleotides. In certain embodiments, the molecules are compounds comprising modified oligonucleotides.
As used herein, “cleavable moiety” means a bond or group of atoms that is cleaved under physiological conditions, for example, inside a cell, a subject, or a human.
As used herein, “complementary” in reference to an oligonucleotide means that at least 70% of the nucleobases of the oligonucleotide or one or more portions thereof and the nucleobases of a target nucleic acid or one or more portions thereof are capable of hydrogen bonding with one another when the nucleobase sequence of the oligonucleotide and the other nucleic acid are aligned in opposing directions. As used herein, complementary nucleobases means nucleobases that are capable of forming hydrogen bonds with one another. Complementary nucleobase pairs include adenine (A) and thymine (T), adenine (A) and uracil (U), cytosine (C) and guanine (G), 5-methyl cytosine (mC) and guanine (G). Complementary oligonucleotides and/or target nucleic acids need not have nucleobase complementarity at each nucleoside. Rather, some mismatches are tolerated. As used herein, “fully complementary” or “100% complementary” in reference to an oligonucleotide, or portion thereof, means that the oligonucleotide, or portion thereof, is complementary to another oligonucleotide or target nucleic acid at each nucleobase of the shorter of the two oligonucleotides, or at each nucleoside if the oligonucleotides are the same length.
As used herein, “conjugate group” means a group of atoms that is directly or indirectly attached to an oligonucleotide. Conjugate groups include a conjugate moiety and a conjugate linker that attaches the conjugate moiety to the oligonucleotide. In certain embodiments, the conjugate moiety comprises a cell-targeting moiety.
As used herein, “conjugate linker” means a single bond or a group of atoms comprising at least one bond that connects a conjugate moiety to an oligonucleotide.
As used herein, “conjugate moiety” means a group of atoms that is attached to an oligonucleotide via a conjugate linker.
As used herein, “constrained ethyl” or “cEt” or “cEt modified sugar moiety” means a β-D ribosyl bicyclic sugar moiety wherein the second ring of the bicyclic sugar is formed via a bridge connecting the 4′-carbon and the 2′-carbon of the β-D ribosyl sugar moiety, wherein the bridge has the formula 4′-CH(CH3)—O-2′, and wherein the methyl group of the bridge is in the S configuration.
As used herein, “contiguous” in the context of an oligonucleotide refers to nucleosides, nucleobases, sugar moieties, or internucleoside linkages that are immediately adjacent to each other. For example, “contiguous nucleobases” means nucleobases that are immediately adjacent to each other in a sequence.
As used herein, “FXII RNA” is the RNA expression product of a human F12 gene.
As used herein, “FXII protein” is the protein expression product of a FXII RNA, including zymogen and activated forms FXII protein.
As used herein, “gapmer” means an oligonucleotide having a central region positioned between a 5′-region and a 3′-region. Each nucleoside of the 5′-region and the 3′-region comprises a modified sugar moiety. The 3′- and 5′-most nucleosides of the central region are 2′-deoxynucleosides. The “central region” may be referred to as a “gap”, and the “5′-region” and “3′-region” may also be referred to as a “wing.” The hybridization of a gapmer to a target nucleic acid results in RNase H mediated cleavage of the target nucleic acid.
As used herein, “hotspot region” is a range of nucleobases on a target nucleic acid that is amenable to oligomeric compound-mediated reduction of the amount or activity of the target nucleic acid.
As used herein, “hybridization” means the pairing or annealing of complementary oligonucleotides and/or nucleic acids. While not limited to a particular mechanism, the most common mechanism of hybridization involves hydrogen bonding, which may be Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding, between complementary nucleobases.
As used herein, “identifying a subject at risk for developing a thromboembolic condition” means identifying a subject having been diagnosed with a thromboembolic condition or identifying a subject that has a risk factor for developing a thromboembolic condition.
As used herein, “internucleoside linkage” means the covalent linkage between contiguous nucleosides in an oligonucleotide. As used herein, “modified internucleoside linkage” means any internucleoside linkage other than a phosphodiester internucleoside linkage. “Phosphorothioate internucleoside linkage” is a modified internucleoside linkage in which one of the non-bridging oxygen atoms of a phosphodiester internucleoside linkage is replaced with a sulfur atom.
As used herein, “linker-nucleoside” means a nucleoside that links, either directly or indirectly, an oligonucleotide to a conjugate moiety. Linker-nucleosides are located within the conjugate linker of an oligomeric compound. Linker-nucleosides are not considered part of the oligonucleotide portion of an oligomeric compound even if they are contiguous with the oligonucleotide.
As used herein, “mismatch” or “non-complementary” means a nucleobase of a first oligonucleotide that is not complementary with the corresponding nucleobase of a second oligonucleotide or target nucleic acid when the first and second oligonucleotide are aligned.
As used herein, “motif” means the pattern of unmodified and/or modified sugar moieties, nucleobases, and/or internucleoside linkages, in an oligonucleotide.
As used herein, “non-bicyclic modified sugar moiety” means a modified sugar moiety that comprises a modification, such as a substituent, that does not form a bridge between two atoms of the sugar to form a second ring.
As used herein, “nucleobase” means an unmodified nucleobase or a modified nucleobase. As used herein an “unmodified nucleobase” is adenine (A), thymine (T), cytosine (C), uracil (U), or guanine (G). As used herein, a “modified nucleobase” is a group of atoms other than unmodified A, T, C, U, or G capable of pairing with at least one unmodified nucleobase. A “5-methyl cytosine” is a modified nucleobase. A universal base is a modified nucleobase that can pair with any one of the five unmodified nucleobases. As used herein, “nucleobase sequence” means the order of contiguous nucleobases in a target nucleic acid or oligonucleotide independent of any sugar or internucleoside linkage modification.
As used herein, “nucleoside” means a compound comprising a nucleobase and a sugar moiety. The nucleobase and sugar moiety are each, independently, unmodified or modified. As used herein, “modified nucleoside” means a nucleoside comprising a modified nucleobase and/or a modified sugar moiety. Modified nucleosides include abasic nucleosides, which lack a nucleobase. “Linked nucleosides” are nucleosides that are connected in a contiguous sequence (i.e., no additional nucleosides are presented between those that are linked).
As used herein, “oligomeric compound” means an oligonucleotide and optionally one or more additional features, such as a conjugate group or terminal group. An oligomeric compound may be paired with a second oligomeric compound that is complementary to the first oligomeric compound or may be unpaired. A “singled-stranded oligomeric compound” is an unpaired oligomeric compound. The term “oligomeric duplex” means a duplex formed by two oligomeric compounds having complementary nucleobase sequences. Each oligomeric compound of an oligomeric duplex may be referred to as a “duplexed oligomeric compound.”
As used herein, “oligonucleotide” means a strand of linked nucleosides connected via internucleoside linkages, wherein each nucleoside and internucleoside linkage may be modified or unmodified. Unless otherwise indicated, oligonucleotides consist of 8-50 linked nucleosides. As used herein, “modified oligonucleotide” means an oligonucleotide, wherein at least one nucleoside or internucleoside linkage is modified. As used herein, “unmodified oligonucleotide” means an oligonucleotide that does not comprise any nucleoside modifications or internucleoside modifications.
As used herein, “pharmaceutically acceptable carrier or diluent” means any substance suitable for use in administering to a subject. Certain such carriers enable pharmaceutical compositions to be formulated as, for example, tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspension and lozenges for the oral ingestion by a subject. In certain embodiments, a pharmaceutically acceptable carrier or diluent is sterile water, sterile saline, or sterile buffer solution. In certain embodiments, a pharmaceutically acceptable carrier or diluent is phosphate buffered saline.
As used herein, “pharmaceutically acceptable salts” means physiologically and pharmaceutically acceptable salts of compounds. Pharmaceutically acceptable salts retain the desired biological activity of the parent compound and do not impart undesired toxicological effects thereto.
As used herein, “pharmaceutical composition” means a mixture of substances suitable for administering to a subject. For example, a pharmaceutical composition may comprise an oligomeric compound and a sterile aqueous solution. In certain embodiments, a pharmaceutical composition shows activity in free uptake assay in certain cell lines.
As used herein, “prevent” or “preventing” refers to delaying or forestalling the onset or development of a thromboembolic condition for a period of time or indefinitely.
As used herein, “prodrug” means a therapeutic agent in a form outside the body that is converted to a different form within a subject or cells thereof. Typically, conversion of a prodrug within the subject is facilitated by the action of an enzymes (e.g., endogenous or viral enzyme) or chemicals present in cells or tissues and/or by physiologic conditions.
As used herein, “reducing the amount or activity,” or “inhibiting the amount or activity,” in connection with a gene transcript (RNA) refers to a reduction or blockade of the transcriptional expression or activity relative to the transcriptional expression or activity in an untreated or control sample and does not necessarily indicate a total elimination of transcriptional expression or activity.
As used herein, “reducing the amount or activity,” or “inhibiting the amount or activity,” in connection with a protein refers to a reduction or blockade of the protein's expression or activity relative to the protein expression or activity in an untreated or control sample and does not necessarily indicate a total elimination of protein expression or activity.
As used herein, “RNA” means an RNA transcript and includes pre-mRNA and mature mRNA unless otherwise specified.
As used herein, “RNAi compound” means an antisense compound that acts, at least in part, through RISC or Ago2 to modulate a target nucleic acid and/or protein encoded by a target nucleic acid. RNAi compounds include, but are not limited to double-stranded siRNA, single-stranded RNA (ssRNA), and microRNA, including microRNA mimics. In certain embodiments, an RNAi compound modulates the amount, activity, and/or splicing of a target nucleic acid. The term RNAi compound excludes antisense compounds that act through RNase H.
As used herein, “self-complementary” in reference to an oligonucleotide means an oligonucleotide that at least partially hybridizes to itself.
As used herein, “standard cell assay” means the assay described in Example 1 and reasonable variations thereof.
As used herein, “stereorandom chiral center” in the context of a population of molecules of identical molecular formula means a chiral center having a random stereochemical configuration. For example, in a population of molecules comprising a stereorandom chiral center, the number of molecules having the (S) configuration of the stereorandom chiral center may be but is not necessarily the same as the number of molecules having the (R) configuration of the stereorandom chiral center. The stereochemical configuration of a chiral center is considered random when it is the result of a synthetic method that is not designed to control the stereochemical configuration. In certain embodiments, a stereorandom chiral center is a stereorandom phosphorothioate internucleoside linkage.
As used herein, “subject” means a human or non-human subject.
As used herein, “sugar moiety” means an unmodified sugar moiety or a modified sugar moiety. As used herein, “unmodified sugar moiety” means a 2′-OH(H) β-D-ribosyl moiety, as found in RNA (an “unmodified RNA sugar moiety”), or a 2′-H(H) β-D-deoxyribosyl sugar moiety, as found in DNA (an “unmodified DNA sugar moiety”). Unmodified sugar moieties have one hydrogen at each of the 1′, 3′, and 4′ positions, an oxygen at the 3′ position, and two hydrogens at the 5′ position. As used herein, “modified sugar moiety” or “modified sugar” means a modified furanosyl sugar moiety or a sugar surrogate.
As used herein, “sugar surrogate” means a modified sugar moiety having other than a furanosyl moiety that can link a nucleobase to another group, such as an internucleoside linkage, conjugate group, or terminal group in an oligonucleotide. Modified nucleosides comprising sugar surrogates can be incorporated into one or more positions within an oligonucleotide and such oligonucleotides are capable of hybridizing to complementary oligomeric compounds or target nucleic acids.
As used herein, “symptom” means any physical feature or test result that indicates the existence or extent of a disease or disorder. In certain embodiments, a symptom is apparent to a subject or to a medical professional examining or testing said subject.
As used herein, “target nucleic acid” and “target RNA” mean a nucleic acid that an antisense compound is designed to affect.
As used herein, “target region” means a portion of a target nucleic acid to which an oligomeric compound is designed to hybridize.
As used herein, “terminal group” means a chemical group or group of atoms that is covalently linked to a terminus of an oligonucleotide.
As used herein, “therapeutically effective amount” means an amount of a pharmaceutical agent that provides a therapeutic benefit to a subject. For example, a therapeutically effective amount improves a symptom of a disease.
As used herein, “thromboembolic condition” means any disease or condition involving a thrombosis or an embolism. A thrombosis is a pathological development of a blood clot, and an embolism occurs when the blood clot migrates to another part of the body and interferes with organ function. Conditions in which a thrombosis and/or an embolism occur are collectively referred to as a thromboembolic condition. Depending on where the blood clot occurs or settles, it may result in a variety of thromboembolic conditions, e.g., deep vein thrombosis (DVT), myocardial infarction (MI), pulmonary embolism (PE), and stroke. In certain embodiments, the thromboembolic condition comprises deep vein thrombosis, venous thrombosis, arterial thrombosis, pulmonary embolism, myocardial infarction, or stroke. Thromboembolic conditions may also be referred to as thromboembolic events or thrombotic events.
Certain Embodiments
The present disclosure provides the following non-limiting numbered embodiments:
Embodiment 1. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to an equal length portion of a FXII RNA, and wherein the modified oligonucleotide comprises at least one modification selected from a modified sugar moiety and a modified internucleoside linkage.
Embodiment 2. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides and having a nucleobase sequence comprising at least 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases of any of SEQ ID NOs: 20-5042.
Embodiment 3. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides and having a nucleobase sequence comprising at least 12, 13, 14, 15, 16, 17, or 18 contiguous nucleobases of any of SEQ ID NOs: 3379 and 5006.
Embodiment 4. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 contiguous nucleobases complementary to: an equal length portion of nucleobases 1,899-1,979 of SEQ ID NO: 1; or an equal length portion of nucleobases 2,004-2,045 of SEQ ID NO: 1.
Embodiment 5. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, or at least 18 contiguous nucleobases of a sequence selected from: SEQ ID NOs: 92, 156, 270, 271, 345, 346, 420, 421, 496, 497, 571, 572, 653, 654, 728, 729, 802, 803, 877, 951, 1028, 1103, 1179, 1253, 1330, 1407, 1484, 1560, 1636, 1710, 1711, 1784, 1859, 1860, 1934, 1935, 2008, 2009, 2084, 2085, 2158, 2232, 2233, 2307, 2308, 2393, 2394, 2395, 2396, 2397, 2398, 2461, 2537, 2538, 2614, 2615, 2691, 2692, 2768, 2769, 2845, 2846, 2921, 2922, 2997, 2998, 3073, 3074, 3144, 3145, 3226, 3227, 3303, 3304, 3379, 3380, 3455, 3456, 3531, 3607, 3683, 3759, 3835, 3911, 4948, 4949, 4954, 4955, 4956, 4957, 4961, 4965, 4966, 4967, 4971, 4972, 4976, 4994, 4995, 4996, 4997, 4998, 4999, 5024, 5025, 5026, 5027, 5028, and 5029; or SEQ ID NOs: 95, 159, 162, 163, 164, 165, 347, 422, 498, 573, 953, 1030, 1105, 1255, 1332, 1409, 1486, 1562, 2309, 2463, 2540, 2617, 2694, 2771, 2848, 2924, 3000, 3076, 3147, 3229, 3306, 3382, 3457, 3533, 3609, 3685, 3761, 3837, 3913, 5000, 5001, 5002, 5003, 5004, 5005, 5006, 5031, 5032, 5033, 5034, 5035, 5036, and 5037.
Embodiment 6. The oligomeric compound of any of embodiments 1-5, wherein the modified oligonucleotide has a nucleobase sequence that is at least 80%, 85%, 90%, 95%, or 100% complementary to the nucleobase sequence of any one of SEQ ID NOS: 1-4 when measured across the entire nucleobase sequence of the modified oligonucleotide.
Embodiment 7. The oligomeric compound of any of embodiments 1-6, wherein the modified oligonucleotide comprises at least one modified nucleoside.
Embodiment 8. The oligomeric compound of embodiment 7, wherein the modified oligonucleotide comprises at least one modified nucleoside comprising a modified sugar moiety.
Embodiment 9. The oligomeric compound of embodiment 8, wherein the modified oligonucleotide comprises at least one modified nucleoside comprising a bicyclic sugar moiety.
Embodiment 10. The oligomeric compound of embodiment 9, wherein the modified oligonucleotide comprises at least one modified nucleoside comprising a bicyclic sugar moiety having a 2′-4′ bridge, wherein the 2′-4′ bridge is selected from —O—CH2—; and —O—CH(CH3)—.
Embodiment 11. The oligomeric compound of any of embodiments 8-10, wherein the modified oligonucleotide comprises at least one modified nucleoside comprising a non-bicyclic modified sugar moiety.
Embodiment 12. The oligomeric compound of embodiment 11, wherein the non-bicyclic modified sugar moiety is a 2′-MOE sugar moiety or 2′-OMe sugar moiety.
Embodiment 13. The oligomeric compound of any of embodiments 7-8, wherein the modified oligonucleotide comprises at least one modified nucleoside comprising a sugar surrogate.
Embodiment 14. The oligomeric compound of any of embodiments 1-13, wherein the modified oligonucleotide is a gapmer.
Embodiment 15. The oligomeric compound of any of embodiments 1-14, wherein the modified oligonucleotide has a sugar motif comprising: a 5′-region consisting of 1-6 linked 5′-region nucleosides; a central region consisting of 6-10 linked central region nucleosides; and a 3′-region consisting of 1-6 linked 3′-region nucleosides, wherein each of the 5′-region nucleosides and each of the 3′-region nucleosides comprises a modified sugar moiety and each of the central region nucleosides comprises a 2′-β-D-deoxyribosyl sugar moiety.
Embodiment 16. The oligomeric compound of embodiment 15, wherein the 5′-region consists of 3 linked 5′-region nucleosides; the central region consists of 10 linked central region nucleosides; and the 3′-region consists of 3 linked 3′-region nucleosides, and wherein each of the 5′-region nucleosides and each of the 3′-region nucleosides comprises a cEt modified sugar moiety.
Embodiment 17. The oligomeric compound of any of embodiments 1-16, wherein the modified oligonucleotide comprises at least one modified internucleoside linkage.
Embodiment 18. The oligomeric compound of embodiment 17, wherein each internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage.
Embodiment 19. The oligomeric compound of embodiment 17 or 18, wherein each internucleoside linkage is a phosphorothioate internucleoside linkage.
Embodiment 20. The oligomeric compound of embodiment 17 or 18, wherein the modified oligonucleotide comprises at least one phosphodiester internucleoside linkage.
Embodiment 21. The oligomeric compound of embodiment 17, wherein each internucleoside linkage is independently selected from a phosphodiester internucleoside linkage and a phosphorothioate internucleoside linkage.
Embodiment 22. The oligomeric compound of any of embodiments 1-21, wherein the modified oligonucleotide comprises a modified nucleobase.
Embodiment 23. The oligomeric compound of embodiment 22, wherein the modified nucleobase is a 5-methyl cytosine.
Embodiment 24. The oligomeric compound of any of embodiments 1-23, wherein the modified oligonucleotide consists of 12-30, 12-22, 12-20,14-18, 14-20, 15-17, 15-25, 16-20, 18-22 or 18-20 linked nucleosides.
Embodiment 25. The oligomeric compound of any of embodiments 1-23, wherein the modified oligonucleotide consists of 16 linked nucleosides.
Embodiment 26. The oligomeric compound of any of embodiments 1-25, consisting of the modified oligonucleotide.
Embodiment 27. The oligomeric compound of any of embodiments 1-25, comprising a conjugate group comprising a conjugate moiety and a conjugate linker.
Embodiment 28. The oligomeric compound of embodiment 27, wherein the conjugate linker consists of a single bond.
Embodiment 29. The oligomeric compound of embodiment 27, wherein the conjugate linker is cleavable.
Embodiment 30. The oligomeric compound of embodiment 27, wherein the conjugate linker comprises 1-3 linker-nucleosides.
Embodiment 31. The oligomeric compound of 27, wherein the oligomeric compound does not comprise a linker-nucleoside.
Embodiment 32. The oligomeric compound of any of embodiments 28-31, wherein the conjugate group is attached to the modified oligonucleotide at the 5′-end of the modified oligonucleotide.
Embodiment 33. The oligomeric compound of any of embodiments 28-31, wherein the conjugate group is attached to the modified oligonucleotide at the 3′-end of the modified oligonucleotide.
Embodiment 34. The oligomeric compound of any of embodiments 1-33, comprising a terminal group.
Embodiment 35. The oligomeric compound of any one of embodiments 1-34, wherein the conjugate group comprises a GalNAc moiety.
Embodiment 36. The oligomeric compound of anyone of embodiments 1-35, wherein the conjugate group comprises a cell-targeting moiety, a conjugate linker and a cleavable moiety, and wherein the cell-targeting moiety and the conjugate linker together have the following chemical structure:
![embedded image]()
Embodiment 37. The oligomeric compound of any of embodiments 1-36, wherein the modified oligonucleotide is a single-stranded modified oligonucleotide.
Embodiment 38. An oligomeric compound according to the following chemical structure:
![embedded image]()
(SEQ ID NO: 3379), or a salt thereof.
Embodiment 39. The oligomeric compound of embodiment 38, which is the sodium salt or the potassium salt.
Embodiment 40. An oligomeric compound according to the following chemical structure:
![embedded image]()
(SEQ ID NO: 3379).
Embodiment 41. An oligomeric compound according to the following chemical structure:
![embedded image]()
![embedded image]()
(SEQ ID NO: 5006), or a salt thereof.
Embodiment 42. The oligomeric compound of embodiment 41, which is the sodium salt or the potassium salt.
Embodiment 43. An oligomeric compound according to the following chemical structure:
![embedded image]()
![embedded image]()
(SEQ ID NO: 5006).
Embodiment 44. An oligomeric compound comprising a modified oligonucleotide according to the following chemical notation: (THA-GalNAc3)o Gks mCks Gks Gds Ads Ads Tds mCds Ads mCds mCds Ads Ads Gks Gks Ak (SEQ ID NO: 3379), wherein:
- A=an adenine nucleobase,
- mC=a 5-methyl cytosine nucleobase,
- G=a guanine nucleobase,
- T=a thymine nucleobase,
- k=a cEt modified sugar moiety,
- d=a 2′-β-D-deoxyribosyl sugar moiety,
- o=a phosphodiester linkage, and
- s=a phosphorothioate internucleoside linkage.
Embodiment 45. An oligomeric compound comprising a modified oligonucleotide according to the following chemical notation: (THA-GalNAc3)o Gks mCks Aks mCds Ums Tds Tds Ads Tds Tds Gds Ads Gds Tks Tks mCk (SEQ ID NO: 5006), wherein: A=an adenine nucleobase, mC=a 5-methyl cytosine nucleobase, G=a guanine nucleobase, T=a thymine nucleobase, U=a uracil nucleobase, k=a cEt modified sugar moiety, d=a 2′-β-D-deoxyribosyl sugar moiety, o=a phosphodiester linkage, and s=a phosphorothioate internucleoside linkage.
Embodiment 46. An oligomeric duplex comprising the oligomeric compound of any of embodiments 1-36.
Embodiment 47. An antisense compound comprising or consisting of an oligomeric compound of any of embodiments 1-45 or an oligomeric duplex of embodiment 46.
Embodiment 48. A pharmaceutical composition comprising an oligomeric compound of any of embodiments 1-45, an oligomeric duplex of embodiment 46, or an antisense compound of embodiment 47; and a pharmaceutically acceptable carrier or diluent.
Embodiment 49. The pharmaceutical composition of embodiment 48, wherein the pharmaceutically acceptable diluent is phosphate buffered saline.
Embodiment 50. The pharmaceutical composition of embodiment 49, wherein the pharmaceutical composition consists essentially of the modified oligonucleotide and phosphate buffered saline.
Embodiment 50. A method comprising administering to a subject a pharmaceutical composition of any of embodiments 48-50.
Embodiment 51. A method of ameliorating or preventing a thromboembolic condition, comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition according to any of embodiments 48-50.
Embodiment 51. The method of embodiment 52, wherein the thromboembolic condition is selected from myocardial infarction (MI), stroke, limb ischemia and necrosis, venous thromboembolism (VTE), deep vein thrombosis (DVT), and pulmonary embolism (PE).
Embodiment 53. The method of any of embodiments 51-53, wherein at least one symptom of the thromboembolic condition is ameliorated.
Embodiment 54. The method of embodiment 54, wherein the symptom is pain, shortness of breath, heart burn, cold sweat, fatigue, lightheadedness, dizziness, swelling, cramping, and death.
Embodiment 55. The method of any one of embodiments 51-55, comprising identifying the subject as having the thromboembolic condition or as having a risk factor for having the thromboembolic condition.
Embodiment 56. The method of embodiment 56, wherein the risk factor is surgery, malignancy, pregnancy, aging, use of oral contraceptives, immobility, sepsis, a mechanical heart valve, valvular heart disease, atrial fibrillation, atherosclerosis, antiphospholipid syndrome, an inherited clotting disorder, or an acquired prothrombotic clotting disorder.
Embodiment 57. A method of ameliorating or preventing hereditary angioedema, comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition according to any of embodiments 48-50.
Embodiment 58. A chirally enriched population of oligomeric compounds of any of embodiments 1-45, wherein the population is enriched for oligomeric compounds comprising at least one particular phosphorothioate internucleoside linkage having a particular stereochemical configuration.
Embodiment 59. The chirally enriched population of embodiment 59, wherein the population is enriched for oligomeric compounds comprising at least one particular phosphorothioate internucleoside linkage having the (Sp) configuration.
Embodiment 60. The chirally enriched population of embodiment 59, wherein the population is enriched for oligomeric compounds comprising at least one particular phosphorothioate internucleoside linkage having the (Rp) configuration.
Embodiment 61. The chirally enriched population of embodiment 59, wherein the population is enriched for oligomeric compounds having a particular, independently selected stereochemical configuration at each phosphorothioate internucleoside linkage.
Embodiment 62. The chirally enriched population of embodiment 62, wherein the population is enriched for oligomeric compounds having the (Sp) configuration at each phosphorothioate internucleoside linkage.
Embodiment 63. The chirally enriched population of embodiment 62, wherein the population is enriched for oligomeric compounds having the (Rp) configuration at each phosphorothioate internucleoside linkage.
Embodiment 64. The chirally enriched population of embodiment 62, wherein the population is enriched for oligomeric compounds having the (Rp) configuration at one particular phosphorothioate internucleoside linkage and the (Sp) configuration at each of the remaining phosphorothioate internucleoside linkages.
Embodiment 65. The chirally enriched population of embodiment 59 or embodiment 62, wherein the population is enriched for oligomeric compounds having at least 3 contiguous phosphorothioate internucleoside linkages in the Sp, Sp, and Rp configurations, in the 5′ to 3′ direction.
Embodiment 66. A chirally enriched population of oligomeric compounds of any of embodiments 1-45, wherein all of the phosphorothioate internucleoside linkages of the oligomeric compounds are stereorandom.
I. Certain Oligonucleotides
In certain embodiments, provided herein are oligomeric compounds comprising oligonucleotides, which consist of linked nucleosides. Oligonucleotides may be unmodified oligonucleotides (RNA or DNA) or may be modified oligonucleotides. Modified oligonucleotides comprise at least one modification relative to unmodified RNA or DNA. That is, modified oligonucleotides comprise at least one modified nucleoside (comprising a modified sugar moiety and/or a modified nucleobase) and/or at least one modified internucleoside linkage.
A. Certain Modified Nucleosides
Modified nucleosides comprise a modified sugar moiety or a modified nucleobase or both a modified sugar moiety and a modified nucleobase.
1. Certain Sugar Moieties
In certain embodiments, modified sugar moieties are non-bicyclic modified sugar moieties. In certain embodiments, modified sugar moieties are bicyclic or tricyclic sugar moieties. In certain embodiments, modified sugar moieties are sugar surrogates. Such sugar surrogates may comprise one or more substitutions corresponding to those of other types of modified sugar moieties.
In certain embodiments, modified sugar moieties are non-bicyclic modified sugar moieties comprising a furanosyl ring with one or more substituent groups none of which bridges two atoms of the furanosyl ring to form a bicyclic structure. Such non bridging substituents may be at any position of the furanosyl, including but not limited to substituents at the 2′, 4′, and/or 5′ positions. In certain embodiments one or more non-bridging substituent of non-bicyclic modified sugar moieties is branched. Examples of 2′-substituent groups suitable for non-bicyclic modified sugar moieties include but are not limited to: 2′-F, 2′-OCH3 (“OMe” or “O-methyl”), and 2′-O(CH2)2OCH3 (“MOE”). In certain embodiments, 2′-substituent groups are selected from among: halo, allyl, amino, azido, SH, CN, OCN, CF3, OCF3, O—C1-C10 alkoxy, O—C1-C10 substituted alkoxy, O—C1-C10 alkyl, O—C1-C10 substituted alkyl, S-alkyl, N(Rm)-alkyl, O-alkenyl, S-alkenyl, N(Rm)-alkenyl, O-alkynyl, S-alkynyl, N(Rm)-alkynyl, O-alkylenyl-O-alkyl, alkynyl, alkaryl, aralkyl, O-alkaryl, O-aralkyl, O(CH2)2SCH3, O(CH2)2ON(Rm)(Rn) or OCH2C(═O)—N(Rm)(Rn), where each Rm and Rn is, independently, H, an amino protecting group, or substituted or unsubstituted C1-C10 alkyl, and the 2′-substituent groups described in Cook et al., U.S. Pat. No. 6,531,584; Cook et al., U.S. Pat. No. 5,859,221; and Cook et al., U.S. Pat. No. 6,005,087. Certain embodiments of these 2′-substituent groups can be further substituted with one or more substituent groups independently selected from among: hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro (NO2), thiol, thioalkoxy, thioalkyl, halogen, alkyl, aryl, alkenyl and alkynyl. Examples of 4′-substituent groups suitable for non-bicyclic modified sugar moieties include but are not limited to alkoxy (e.g., methoxy), alkyl, and those described in Manoharan et al., WO 2015/106128. Examples of 5′-substituent groups suitable for non-bicyclic modified sugar moieties include but are not limited to: 5-methyl (R or S), 5′-vinyl, and 5′-methoxy. In certain embodiments, non-bicyclic modified sugar moieties comprise more than one non-bridging sugar substituent, for example, 2′-F-5′-methyl sugar moieties and the modified sugar moieties and modified nucleosides described in Migawa et al., WO 2008/101157 and Rajeev et al., US2013/0203836.
In certain embodiments, a 2′-substituted non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2′-substituent group selected from: F, NH2, N3, OCF3, OCH3, O(CH2)3NH2, CH2CH═CH2, OCH2CH═CH2, OCH2CH2OCH3, O(CH2)2SCH3, O(CH2)2ON(Rm)(Rn), O(CH2)2O(CH2)2N(CH3)2, and N-substituted acetamide (OCH2C(═O)—N(Rm)(Rn)), where each Rm and Rn is, independently, H, an amino protecting group, or substituted or unsubstituted C1-C10 alkyl.
In certain embodiments, a 2′-substituted nucleoside non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2′-substituent group selected from: F, OCF3, OCH3, OCH2CH2OCH3, O(CH2)2SCH3, O(CH2)2ON(CH3)2, O(CH2)2O(CH2)2N(CH3)2, and OCH2C(═O)—N(H)CH3 (“NMA”).
In certain embodiments, a 2′-substituted non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2′-substituent group selected from: F, OCH3, and OCH2CH2OCH3.
Certain modified sugar moieties comprise a substituent that bridges two atoms of the furanosyl ring to form a second ring, resulting in a bicyclic sugar moiety. In certain such embodiments, the bicyclic sugar moiety comprises a bridge between the 4′ and the 2′ furanose ring atoms. Examples of such 4′ to 2′ bridging sugar substituents include but are not limited to: 4′-CH2-2′, 4′-(CH2)2-2′, 4′-(CH2)3-2′, 4′-CH2—O-2′ (“LNA”), 4′-CH2—S-2′, 4′-(CH2)2—O-2′ (“ENA”), 4′-CH(CH3)—O-2′ (referred to as “constrained ethyl” or “cEt”), 4′-CH2—O—CH2-2′, 4′-CH2—N(R)-2′, 4′-CH(CH2OCH3)—O-2′ (“constrained MOE” or “cMOE”) and analogs thereof (see, e.g., Seth et al., U.S. Pat. No. 7,399,845, Bhat et al., U.S. Pat. No. 7,569,686, Swayze et al., U.S. Pat. No. 7,741,457, and Swayze et al., U.S. Pat. No. 8,022,193), 4′-C(CH3)(CH3)—O-2′ and analogs thereof (see, e.g., Seth et al., U.S. Pat. No. 8,278,283), 4′-CH2—N(OCH3)-2′ and analogs thereof (see, e.g., Prakash et al., U.S. Pat. No. 8,278,425), 4′-CH2—O—N(CH3)-2′ (see, e.g., Allerson et al., U.S. Pat. No. 7,696,345 and Allerson et al., U.S. Pat. No. 8,124,745), 4′-CH2—C(H)(CH3)-2′ (see, e.g., Zhou, et al., J. Org. Chem., 2009, 74, 118-134), 4′-CH2—C(═CH2)-2′ and analogs thereof (see e.g., Seth et al., U.S. Pat. No. 8,278,426), 4′-C(RaRb)—N(R)—O-2′, 4′-C(RaRb)—O—N(R)-2′, 4′-CH2—O—N(R)-2′, and 4′-CH2—N(R)—O- 2′, wherein each R, Ra, and Rb is, independently, H, a protecting group, or C1-C12 alkyl (see, e.g. Imanishi et al., U.S. Pat. No. 7,427,672).
In certain embodiments, such 4′ to 2′ bridges independently comprise from 1 to 4 linked groups independently selected from: —[C(Ra)(Rb)]n—, —[C(Ra)(Rb)]n—O—, —C(Ra)═C(Rb)—, —C(Ra)═N—, —C(═NRa)—, —C(═O)—, —C(═S)—, —O—, —Si(Ra)2—, —S(═O)x—, and —N(Ra)—;
- wherein: x is 0, 1, or 2; n is 1, 2, 3, or 4; each Ra and Rb is, independently, H, a protecting group, hydroxyl, C1-C12 alkyl, substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkynyl, C5-C20 aryl, substituted C5-C20 aryl, heterocycle radical, substituted heterocycle radical, heteroaryl, substituted heteroaryl, C5-C7 alicyclic radical, substituted C5-C7 alicyclic radical, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(═O)—H), substituted acyl, CN, sulfonyl (S(═O)2-J1), or sulfoxyl (S(═O)-J1); and
- each J1 and J2 is, independently, H, C1-C12 alkyl, substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkynyl, C5-C20 aryl, substituted C5-C20 aryl, acyl (C(═O)—H), substituted acyl, a heterocycle radical, a substituted heterocycle radical, C1-C12 aminoalkyl, substituted C1-C12 aminoalkyl, or a protecting group.
Additional bicyclic sugar moieties are known in the art, see, for example: Freier et al., Nucleic Acids Research, 1997, 25(22), 4429-4443, Albaek et al., J. Org. Chem., 2006, 71, 7731-7740, Singh et al., Chem. Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54, 3607-3630; Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222; Singh et al., J. Org. Chem., 1998, 63, 10035-10039; Srivastava et al., J. Am. Chem. Soc., 2007, 129, 8362-8379; Wengel et a., U.S. Pat. No. 7,053,207; Imanishi et al., U.S. Pat. No. 6,268,490; Imanishi et al. U.S. Pat. No. 6,770,748; Imanishi et al., U.S. RE44,779; Wengel et al., U.S. Pat. No. 6,794,499; Wengel et al., U.S. Pat. No. 6,670,461; Wengel et al., U.S. Pat. No. 7,034,133; Wengel et al., U.S. Pat. No. 8,080,644; Wengel et al., U.S. Pat. No. 8,034,909; Wengel et al., U.S. Pat. No. 8,153,365; Wengel et al., U.S. Pat. No. 7,572,582; Ramasamy et al., U.S. Pat. No. 6,525,191; Torsten et al., WO 2004/106356; Wengel et al., WO 1999/014226; Seth et al., WO 2007/134181; Seth et al., U.S. Pat. No. 7,547,684; Seth et al., U.S. Pat. No. 7,666,854; Seth et al., U.S. Pat. No. 8,088,746; Seth et al., U.S. Pat. No. 7,750,131; Seth et al., U.S. Pat. No. 8,030,467; Seth et al., U.S. Pat. No. 8,268,980; Seth et al., U.S. Pat. No. 8,546,556; Seth et al., U.S. Pat. No. 8,530,640; Migawa et al., U.S. Pat. No. 9,012,421; Seth et al., U.S. Pat. No. 8,501,805; and U.S. Patent Publication Nos. Allerson et al., US2008/0039618 and Migawa et al., US2015/0191727.
In certain embodiments, bicyclic sugar moieties and nucleosides incorporating such bicyclic sugar moieties are further defined by isomeric configuration. For example, an LNA nucleoside (described herein) may be in the α-L configuration or in the β-D configuration.
![embedded image]()
α-L-methyleneoxy (4′-CH2—O-2′) or α-L-LNA bicyclic nucleosides have been incorporated into oligonucleotides that showed antisense activity (Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372). Herein, general descriptions of bicyclic nucleosides include both isomeric configurations. When the positions of specific bicyclic nucleosides (e.g., LNA or cEt) are identified in exemplified embodiments herein, they are in the β-D configuration, unless otherwise specified.
In certain embodiments, modified sugar moieties comprise one or more non-bridging sugar substituent and one or more bridging sugar substituent (e.g., 5′-substituted and 4′-2′ bridged sugars).
In certain embodiments, modified sugar moieties are sugar surrogates. In certain such embodiments, the oxygen atom of the sugar moiety is replaced, e.g., with a sulfur, carbon or nitrogen atom. In certain such embodiments, such modified sugar moieties also comprise bridging and/or non-bridging substituents as described herein. For example, certain sugar surrogates comprise a 4′-sulfur atom and a substitution at the 2′-position (see, e.g., Bhat et al., U.S. Pat. No. 7,875,733 and Bhat et al., U.S. Pat. No. 7,939,677) and/or the 5′ position.
In certain embodiments, sugar surrogates comprise rings having other than 5 atoms. For example, in certain embodiments, a sugar surrogate comprises a six-membered tetrahydropyran (“THP”). Such tetrahydropyrans may be further modified or substituted. Nucleosides comprising such modified tetrahydropyrans include but are not limited to hexitol nucleic acid (“HNA”), anitol nucleic acid (“ANA”), manitol nucleic acid (“MNA”) (see, e.g., Leumann, C J. Bioorg. & Med. Chem. 2002, 10, 841-854), fluoro HNA:
![embedded image]()
(“F-HNA”, see e.g. Swayze et al., U.S. Pat. No. 8,088,904; Swayze et al., U.S. Pat. No. 8,440,803; Swayze et al., U.S. Pat. No. 8,796,437; and Swayze et al., U.S. Pat. No. 9,005,906; F-HNA can also be referred to as a F-THP or 3′-fluoro tetrahydropyran), and nucleosides comprising additional modified THP compounds having the formula:
![embedded image]()
wherein, independently, for each of said modified THP nucleoside:
- Bx is a nucleobase moiety;
- T3 and T4 are each, independently, an internucleoside linking group linking the modified THP nucleoside to the remainder of an oligonucleotide or one of T3 and T4 is an internucleoside linking group linking the modified THP nucleoside to the remainder of an oligonucleotide and the other of T3 and T4 is H, a hydroxyl protecting group, a linked conjugate group, or a 5′ or 3′-terminal group;
q1, q2, q3, q4, q5, q6 and q7 are each, independently, H, C1-C6 alkyl, substituted C1-C6 alkyl, C2-C6 alkenyl, substituted C2-C6 alkenyl, C2-C6 alkynyl, or substituted C2-C6 alkynyl; and
- each of R1 and R2 is independently selected from among: hydrogen, halogen, substituted or unsubstituted alkoxy, NJ1J2, SJ1, N3, OC(═X)J1, OC(═X)NJ1J2, NJ3C(═X)NJ1J2, and CN, wherein X is O, S or NJ1, and each J1, J2, and J3 is, independently, H or C1-C6 alkyl.
In certain embodiments, modified THP nucleosides are provided wherein q1, q2, q3, q4, q5, q6 and q7 are each H. In certain embodiments, at least one of q1, q2, q3, q4, q5, q6 and q7 is other than H. In certain embodiments, at least one of q1, q2, q3, q4, q5, q6 and q7 is methyl. In certain embodiments, modified THP nucleosides are provided wherein one of R1 and R2 is F. In certain embodiments, R1 is F and R2 is H, in certain embodiments, R1 is methoxy and R2 is H, and in certain embodiments, R1 is methoxyethoxy and R2 is H.
In certain embodiments, sugar surrogates comprise rings having more than 5 atoms and more than one heteroatom. For example, nucleosides comprising morpholino sugar moieties and their use in oligonucleotides have been reported (see, e.g., Braasch et al., Biochemistry, 2002, 41, 4503-4510 and Summerton et al., U.S. Pat. No. 5,698,685; Summerton et al., U.S. Pat. No. 5,166,315; Summerton et al., U.S. Pat. No. 5,185,444; and Summerton et al., U.S. Pat. No. 5,034,506). As used here, the term “morpholino” means a sugar surrogate having the following structure:
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In certain embodiments, morpholinos may be modified, for example by adding or altering various substituent groups from the above morpholino structure. Such sugar surrogates are referred to herein as “modified morpholinos.”
In certain embodiments, sugar surrogates comprise acyclic moieties. Examples of nucleosides and oligonucleotides comprising such acyclic sugar surrogates include but are not limited to: peptide nucleic acid (“PNA”), acyclic butyl nucleic acid (see, e.g., Kumar et al., Org. Biomol. Chem., 2013, 11, 5853-5865), and nucleosides and oligonucleotides described in Manoharan et al., WO2011/133876.
Many other bicyclic and tricyclic sugar and sugar surrogate ring systems are known in the art that can be used in modified nucleosides.
2. Certain Modified Nucleobases
In certain embodiments, modified oligonucleotides comprise one or more nucleosides comprising an unmodified nucleobase. In certain embodiments, modified oligonucleotides comprise one or more nucleosides comprising a modified nucleobase. In certain embodiments, modified oligonucleotides comprise one or more nucleosides that does not comprise a nucleobase, referred to as an abasic nucleoside.
In certain embodiments, modified nucleobases are selected from: 5-substituted pyrimidines, 6-azapyrimidines, alkyl or alkynyl substituted pyrimidines, alkyl substituted purines, and N-2, N-6 and O-6 substituted purines. In certain embodiments, modified nucleobases are selected from: 2-aminopropyladenine, 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-N-methylguanine, 6-N-methyladenine, 2-propyladenine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-propynyl (—C≡C—CH3) uracil, 5-propynylcytosine, 6-azouracil, 6-azocytosine, 6-azothymine, 5-ribosyluracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl, 8-aza and other 8-substituted purines, 5-halo, particularly 5-bromo, 5-trifluoromethyl, 5-halouracil, and 5-halocytosine, 7-methylguanine, 7-methyladenine, 2-F-adenine, 2-aminoadenine, 7-deazaguanine, 7-deazaadenine, 3-deazaguanine, 3-deazaadenine, 6-N-benzoyladenine, 2-N-isobutyrylguanine, 4-N-benzoylcytosine, 4-N-benzoyluracil, 5-methyl 4-N-benzoylcytosine, 5-methyl 4-N-benzoyluracil, universal bases, hydrophobic bases, promiscuous bases, size-expanded bases, and fluorinated bases. Further modified nucleobases include tricyclic pyrimidines, such as 1,3-diazaphenoxazine-2-one, 1,3-diazaphenothiazine-2-one and 9-(2-aminoethoxy)-1,3-diazaphenoxazine-2-one (G-clamp). Modified nucleobases may also include those in which the purine or pyrimidine base is replaced with other heterocycles, for example 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine and 2-pyridone. Further nucleobases include those disclosed in Merigan et al., U.S. Pat. No. 3,687,808, those disclosed in The Concise Encyclopedia Of Polymer Science And Engineering, Kroschwitz, J. I., Ed., John Wiley & Sons, 1990, 858-859; Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613; Sanghvi, Y. S., Chapter 15, Antisense Research and Applications, Crooke, S. T. and Lebleu, B., Eds., CRC Press, 1993, 273-288; and those disclosed in Chapters 6 and 15, Antisense Drug Technology, Crooke S. T., Ed., CRC Press, 2008, 163-166 and 442-443.
Publications that teach the preparation of certain of the above noted modified nucleobases as well as other modified nucleobases include without limitation, Manoharan et al., US2003/0158403; Manoharan et al., US2003/0175906; Dinh et al., U.S. Pat. No. 4,845,205; Spielvogel et al., U.S. Pat. No. 5,130,302; Rogers et al., U.S. Pat. No. 5,134,066; Bischofberger et al., U.S. Pat. No. 5,175,273; Urdea et al., U.S. Pat. No. 5,367,066; Benner et al., U.S. Pat. No. 5,432,272; Matteucci et al., U.S. Pat. No. 5,434,257; Gmeiner et al., U.S. Pat. No. 5,457,187; Cook et al., U.S. Pat. No. 5,459,255; Froehler et al., U.S. Pat. No. 5,484,908; Matteucci et al., U.S. Pat. No. 5,502,177; Hawkins et al., U.S. Pat. No. 5,525,711; Haralambidis et al., U.S. Pat. No. 5,552,540; Cook et al., U.S. Pat. No. 5,587,469; Froehler et al., U.S. Pat. No. 5,594,121; Switzer et al., U.S. Pat. No. 5,596,091; Cook et al., U.S. Pat. No. 5,614,617; Froehler et al., U.S. Pat. No. 5,645,985; Cook et al., U.S. Pat. No. 5,681,941; Cook et al., U.S. Pat. No. 5,811,534; Cook et al., U.S. Pat. No. 5,750,692; Cook et al., U.S. Pat. No. 5,948,903; Cook et al., U.S. Pat. No. 5,587,470; Cook et al., U.S. Pat. No. 5,457,191; Matteucci et al., U.S. Pat. No. 5,763,588; Froehler et al., U.S. Pat. No. 5,830,653; Cook et al., U.S. Pat. No. 5,808,027; Cook et al., 6,166,199; and Matteucci et al., U.S. Pat. No. 6,005,096.
3. Certain Modified Internucleoside Linkages
In certain embodiments, nucleosides of modified oligonucleotides may be linked together using any internucleoside linkage. The two main classes of internucleoside linking groups are defined by the presence or absence of a phosphorus atom. Representative phosphorus-containing internucleoside linkages include but are not limited to phosphodiesters, which contain a phosphodiester bond (“P(O2)═O”) (also referred to as unmodified or naturally occurring linkages), phosphotriesters, methylphosphonates, phosphoramidates, and phosphorothioates, and phosphorodithioates. Representative non-phosphorus containing internucleoside linking groups include but are not limited to methylenemethylimino (—CH2—N(CH3)—O—CH2—), thiodiester, thionocarbamate (—O—C(═O)(NH)—S—); siloxane (—O—SiH2—O—); and N,N′-dimethylhydrazine (—CH2—N(CH3)—N(CH3)—). Modified internucleoside linkages, compared to naturally occurring phosphate linkages, can be used to alter, typically increase, nuclease resistance of the oligonucleotide. In certain embodiments, internucleoside linkages having a chiral atom can be prepared as a racemic mixture, or as separate enantiomers. Methods of preparation of phosphorous-containing and non-phosphorous-containing internucleoside linkages are well known to those skilled in the art.
Representative internucleoside linkages having a chiral center include but are not limited to alkylphosphonates and phosphorothioates. Modified oligonucleotides comprising internucleoside linkages having a chiral center can be prepared as populations of modified oligonucleotides comprising stereorandom internucleoside linkages, or as populations of modified oligonucleotides comprising phosphorothioate linkages in particular stereochemical configurations. In certain embodiments, populations of modified oligonucleotides comprise phosphorothioate internucleoside linkages wherein all of the phosphorothioate internucleoside linkages are stereorandom. Such modified oligonucleotides can be generated using synthetic methods that result in random selection of the stereochemical configuration of each phosphorothioate linkage. Nonetheless, as is well understood by those of skill in the art, each individual phosphorothioate of each individual oligonucleotide molecule has a defined stereoconfiguration. In certain embodiments, populations of modified oligonucleotides are enriched for modified oligonucleotides comprising one or more particular phosphorothioate internucleoside linkages in a particular, independently selected stereochemical configuration. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 65% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 70% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 80% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 90% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 99% of the molecules in the population. Such chirally enriched populations of modified oligonucleotides can be generated using synthetic methods known in the art, e.g., methods described in Oka et al., JACS 125, 8307 (2003), Wan et al. Nuc. Acid. Res. 42, 13456 (2014), and WO 2017/015555. In certain embodiments, a population of modified oligonucleotides is enriched for modified oligonucleotides having at least one indicated phosphorothioate in the (Sp) configuration. In certain embodiments, a population of modified oligonucleotides is enriched for modified oligonucleotides having at least one phosphorothioate in the (Rp) configuration. In certain embodiments, modified oligonucleotides comprising (Rp) and/or (Sp) phosphorothioates comprise one or more of the following formulas, respectively, wherein “B” indicates a nucleobase:
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Unless otherwise indicated, chiral internucleoside linkages of modified oligonucleotides described herein can be stereorandom or in a particular stereochemical configuration.
Neutral internucleoside linkages include, without limitation, phosphotriesters, methylphosphonates, MMI (3′-CH2—N(CH3)—O-5′), amide-3 (3′-CH2—C(═O)—N(H)-5′), amide-4 (3′-CH2—N(H)—C(═O)-5′), formacetal (3′-O—CH2—O-5′), methoxypropyl (MOP), and thioformacetal (3′-S—CH2—O-5′). Further neutral internucleoside linkages include nonionic linkages comprising siloxane (dialkylsiloxane), carboxylate ester, carboxamide, sulfide, sulfonate ester and amides (See for example: Carbohydrate Modifications in Antisense Research; Y. S. Sanghvi and P. D. Cook, Eds., ACS Symposium Series 580; Chapters 3 and 4, 40-65). Further neutral internucleoside linkages include nonionic linkages comprising mixed N, O, S and CH2 component parts.
B. Certain Motifs
In certain embodiments, modified oligonucleotides comprise one or more modified nucleosides comprising a modified sugar moiety. In certain embodiments, modified oligonucleotides comprise one or more modified nucleosides comprising a modified nucleobase. In certain embodiments, modified oligonucleotides comprise one or more modified internucleoside linkage. In such embodiments, the modified, unmodified, and differently modified sugar moieties, nucleobases, and/or internucleoside linkages of a modified oligonucleotide define a pattern or motif. In certain embodiments, the patterns of sugar moieties, nucleobases, and internucleoside linkages are each independent of one another. Thus, a modified oligonucleotide may be described by its sugar motif, nucleobase motif and/or internucleoside linkage motif (as used herein, nucleobase motif describes the modifications to the nucleobases independent of the sequence of nucleobases).
1. Certain Sugar Motifs
In certain embodiments, oligonucleotides comprise one or more type of modified sugar and/or unmodified sugar moiety arranged along the oligonucleotide or portion thereof in a defined pattern or sugar motif. In certain instances, such sugar motifs include but are not limited to any of the sugar modifications discussed herein.
In certain embodiments, modified oligonucleotides have a gapmer motif, which is defined by two external regions or “wings” and a central or internal region or “gap.” The three regions of a gapmer motif (the 5′-wing, the gap, and the 3′-wing) form a contiguous sequence of nucleosides wherein at least some of the sugar moieties of the nucleosides of each of the wings differ from at least some of the sugar moieties of the nucleosides of the gap. Specifically, at least the sugar moieties of the nucleosides of each wing that are closest to the gap (the 3′-most nucleoside of the 5′-wing and the 5′-most nucleoside of the 3′-wing) differ from the sugar moiety of the neighboring gap nucleosides, thus defining the boundary between the wings and the gap (i.e., the wing/gap junction). In certain embodiments, the sugar moieties within the gap are the same as one another. In certain embodiments, the gap includes one or more nucleoside having a sugar moiety that differs from the sugar moiety of one or more other nucleosides of the gap. In certain embodiments, the sugar motifs of the two wings are the same as one another (symmetric gapmer). In certain embodiments, the sugar motif of the 5′-wing differs from the sugar motif of the 3′-wing (asymmetric gapmer).
In certain embodiments, the wings of a gapmer comprise 1-6 nucleosides. In certain embodiments, each nucleoside of each wing of a gapmer comprises a modified sugar moiety. In certain embodiments, at least one nucleoside of each wing of a gapmer comprises a modified sugar moiety. In certain embodiments, at least two nucleosides of each wing of a gapmer comprises a modified sugar moiety. In certain embodiments, at least three nucleosides of each wing of a gapmer comprises a modified sugar moiety. In certain embodiments, at least four nucleosides of each wing of a gapmer comprises a modified sugar moiety. In certain embodiments, at least five nucleosides of each wing of a gapmer comprises a modified sugar moiety.
In certain embodiments, the gap of a gapmer comprises 7-12 nucleosides. In certain embodiments, each nucleoside of the gap of a gapmer comprises a 2′-β-D-deoxyribosyl sugar moiety. In certain embodiments, at least one nucleoside of the gap of a gapmer comprises a modified sugar moiety.
In certain embodiments, the gapmer is a deoxy gapmer. In certain embodiments, the nucleosides on the gap side of each wing/gap junction comprise 2′-β-D-deoxyribosyl sugar moieties and the nucleosides on the wing sides of each wing/gap junction comprise modified sugar moieties. In certain embodiments, each nucleoside of the gap comprises a 2′-β-D-deoxyribosyl sugar moiety. In certain embodiments, each nucleoside of each wing of a gapmer comprises a modified sugar moiety. In certain embodiments, exactly one nucleoside of the gap comprises a modified sugar moiety and each remaining nucleoside of the gap comprises a 2′-β-D-deoxyribosyl sugar moiety.
In certain embodiments, modified oligonucleotides comprise or consist of a portion having a fully modified sugar motif. In such embodiments, each nucleoside of the fully modified portion of the modified oligonucleotide comprises a modified sugar moiety. In certain embodiments, each nucleoside of the entire modified oligonucleotide comprises a modified sugar moiety. In certain embodiments, modified oligonucleotides comprise or consist of a portion having a fully modified sugar motif, wherein each nucleoside within the fully modified portion comprises the same modified sugar moiety, referred to herein as a uniformly modified sugar motif. In certain embodiments, a fully modified oligonucleotide is a uniformly modified oligonucleotide. In certain embodiments, each nucleoside of a uniformly modified comprises the same 2′-modification.
Herein, the lengths (number of nucleosides) of the three regions of a gapmer may be provided using the notation [# of nucleosides in the 5′-wing]-[# of nucleosides in the gap]-[# of nucleosides in the 3′-wing]. Thus, a 3-10-3 gapmer consists of 3 linked nucleosides in each wing and 10 linked nucleosides in the gap. Where such nomenclature is followed by a specific modification, that modification is the modification in each sugar moiety of each wing and the gap nucleosides comprise 2′-β-D-deoxyribosyl sugar moieties. Thus, a 5-10-5 MOE gapmer consists of 5 linked 2′-MOE nucleosides in the 5′-wing, 10 linked 2′-β-D-deoxynucleosides in the gap, and 5 linked 2′-MOE nucleosides in the 3′-wing. A 3-10-3 cEt gapmer consists of 3 linked cEt nucleosides in the 5′-wing, 10 linked 2′-β-D-deoxynucleosides in the gap, and 3 linked cEt nucleosides in the 3′-wing. A 5-8-5 gapmer consists of 5 linked nucleosides comprising a modified sugar moiety in the 5′-wing, 8 linked 2′-β-D-deoxynucleosides in the gap, and 5 linked nucleosides comprising a modified sugar moiety in the 3′-wing. A 5-8-5 mixed gapmer has at least two different modified sugar moieties in the 5′- and/or the 3′-wing.
In certain embodiments, modified oligonucleotides are 5-10-5 MOE gapmers. In certain embodiments, modified oligonucleotides are 4-10-6 MOE gapmers. In certain embodiments, modified oligonucleotides are 6-10-4 MOE gapmers. In certain embodiments, modified oligonucleotides are 5-8-5 MOE gapmers. In certain embodiments, modified oligonucleotides are 6-8-4 MOE gapmers. In certain embodiments, modified oligonucleotides are 4-8-6 MOE gapmers. In certain embodiments, modified oligonucleotides are X-Y-Z MOE gapmers, wherein X and Z are independently selected from 1, 2, 3, 4, 5, or 6 and Y is 7, 8, 9, 10, or 11.
2. Certain Nucleobase Motifs
In certain embodiments, oligonucleotides comprise modified and/or unmodified nucleobases arranged along the oligonucleotide or portion thereof in a defined pattern or motif. In certain embodiments, each nucleobase is modified. In certain embodiments, none of the nucleobases are modified. In certain embodiments, each purine or each pyrimidine is modified. In certain embodiments, each adenine is modified. In certain embodiments, each guanine is modified. In certain embodiments, each thymine is modified. In certain embodiments, each uracil is modified. In certain embodiments, each cytosine is modified. In certain embodiments, some or all of the cytosine nucleobases in a modified oligonucleotide are 5-methyl cytosines. In certain embodiments, all of the cytosine nucleobases are 5-methyl cytosines and all of the other nucleobases of the modified oligonucleotide are unmodified nucleobases.
In certain embodiments, modified oligonucleotides comprise a block of modified nucleobases. In certain such embodiments, the block is at the 3′-end of the oligonucleotide. In certain embodiments the block is within 3 nucleosides of the 3′-end of the oligonucleotide. In certain embodiments, the block is at the 5′-end of the oligonucleotide. In certain embodiments the block is within 3 nucleosides of the 5′-end of the oligonucleotide.
In certain embodiments, oligonucleotides having a gapmer motif comprise a nucleoside comprising a modified nucleobase. In certain such embodiments, one nucleoside comprising a modified nucleobase is in the central gap of an oligonucleotide having a gapmer motif. In certain such embodiments, the sugar moiety of said nucleoside is a 2′-β-D-deoxyribosyl sugar moiety. In certain embodiments, the modified nucleobase is selected from: a 2-thiopyrimidine and a 5-propynepyrimidine.
3. Certain Internucleoside Linkage Motifs
In certain embodiments, oligonucleotides comprise modified and/or unmodified internucleoside linkages arranged along the oligonucleotide or portion thereof in a defined pattern or motif. In certain embodiments, each internucleoside linking group is a phosphodiester internucleoside linkage (P(O2)═O). In certain embodiments, each internucleoside linking group of a modified oligonucleotide is a phosphorothioate internucleoside linkage (P(O2)═S). In certain embodiments, each internucleoside linkage of a modified oligonucleotide is independently selected from a phosphorothioate internucleoside linkage and phosphodiester internucleoside linkage. In certain embodiments, each phosphorothioate internucleoside linkage is independently selected from a stereorandom phosphorothioate, a (Sp) phosphorothioate, and a (Rp) phosphorothioate. In certain embodiments, the sugar motif of a modified oligonucleotide is a gapmer and the internucleoside linkages within the gap are all modified. In certain such embodiments, some or all of the internucleoside linkages in the wings are unmodified phosphodiester internucleoside linkages. In certain embodiments, the terminal internucleoside linkages are modified. In certain embodiments, the sugar motif of a modified oligonucleotide is a gapmer, and the internucleoside linkage motif comprises at least one phosphodiester internucleoside linkage in at least one wing, wherein the at least one phosphodiester linkage is not a terminal internucleoside linkage, and the remaining internucleoside linkages are phosphorothioate internucleoside linkages. In certain such embodiments, all of the phosphorothioate linkages are stereorandom. In certain embodiments, all of the phosphorothioate linkages in the wings are (Sp) phosphorothioates, and the gap comprises at least one Sp, Sp, Rp motif. In certain embodiments, populations of modified oligonucleotides are enriched for modified oligonucleotides comprising such internucleoside linkage motifs.
In certain embodiments, modified nucleotides have an internucleoside linkage motif of soooossssssssssooss, wherein each “s” represents a phosphorothioate internucleoside linkage and each “o” represents a phosphodiester internucleoside linkage. In certain embodiments, modified nucleotides have an internucleoside linkage motif of sooooossssssssssoss, wherein each “s” represents a phosphorothioate internucleoside linkage and each “o” represents a phosphodiester internucleoside linkage. In certain embodiments, modified nucleotides have an internucleoside linkage motif of soooosssssssssoss, wherein each “s” represents a phosphorothioate internucleoside linkage and each “o” represents a phosphodiester internucleoside linkage. In certain embodiments, modified nucleotides have an internucleoside linkage motif of sooosssssssssooss, wherein each “s” represents a phosphorothioate internucleoside linkage and each “o” represents a phosphodiester internucleoside linkage. In certain embodiments, modified nucleotides have an internucleoside linkage motif of sooossssssssssoooss, wherein each “s” represents a phosphorothioate internucleoside linkage and each “o” represents a phosphodiester internucleoside linkage. In certain embodiments, modified nucleotides have an internucleoside linkage motif of soosssssssssoooss, wherein each “s” represents a phosphorothioate internucleoside linkage and each “o” represents a phosphodiester internucleoside linkage.
C. Certain Lengths
It is possible to increase or decrease the length of an oligonucleotide without eliminating activity. For example, in Woolf et al. (Proc. Natl. Acad. Sci. USA 89:7305-7309, 1992), a series of oligonucleotides 13-25 nucleobases in length were tested for their ability to induce cleavage of a target RNA in an oocyte injection model. Oligonucleotides 25 nucleobases in length with 8 or 11 mismatch bases near the ends of the oligonucleotides were able to direct specific cleavage of the target RNA, albeit to a lesser extent than the oligonucleotides that contained no mismatches. Similarly, target specific cleavage was achieved using 13 nucleobase oligonucleotides, including those with 1 or 3 mismatches.
In certain embodiments, oligonucleotides (including modified oligonucleotides) can have any of a variety of ranges of lengths. In certain embodiments, oligonucleotides consist of X to Y linked nucleosides, where X represents the fewest number of nucleosides in the range and Y represents the largest number nucleosides in the range. In certain such embodiments, X and Y are each independently selected from 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50; provided that X≤Y. For example, in certain embodiments, oligonucleotides consist of 12 to 13, 12 to 14, 12 to 15, 12 to 16, 12 to 17, 12 to 18, 12 to 19, 12 to 20, 12 to 21, 12 to 22, 12 to 23, 12 to 24, 12 to 25, 12 to 26, 12 to 27, 12 to 28, 12 to 29, 12 to 30, 13 to 14, 13 to 15, 13 to 16, 13 to 17, 13 to 18, 13 to 19, 13 to 20, 13 to 21, 13 to 22, 13 to 23, 13 to 24, 13 to 25, 13 to 26, 13 to 27, 13 to 28, 13 to 29, 13 to 30, 14 to 15, 14 to 16, 14 to 17, 14 to 18, 14 to 19, 14 to 20, 14 to 21, 14 to 22, 14 to 23, 14 to 24, 14 to 25, 14 to 26, 14 to 27, 14 to 28, 14 to 29, 14 to 30, 15 to 16, 15 to 17, 15 to 18, 15 to 19, 15 to 20, 15 to 21, 15 to 22, 15 to 23, 15 to 24, 15 to 25, 15 to 26, 15 to 27, 15 to 28, 15 to 29, 15 to 30, 16 to 17, 16 to 18, 16 to 19, 16 to 20, 16 to 21, 16 to 22, 16 to 23, 16 to 24, 16 to 25, 16 to 26, 16 to 27, 16 to 28, 16 to 29, 16 to 30, 17 to 18, 17 to 19, 17 to 20, 17 to 21, 17 to 22, 17 to 23, 17 to 24, 17 to 25, 17 to 26, 17 to 27, 17 to 28, 17 to 29, 17 to 30, 18 to 19, 18 to 20, 18 to 21, 18 to 22, 18 to 23, 18 to 24, 18 to 25, 18 to 26, 18 to 27, 18 to 28, 18 to 29, 18 to 30, 19 to 20, 19 to 21, 19 to 22, 19 to 23, 19 to 24, 19 to 25, 19 to 26, 19 to 29, 19 to 28, 19 to 29, 19 to 30, 20 to 21, 20 to 22, 20 to 23, 20 to 24, 20 to 25, 20 to 26, 20 to 27, 20 to 28, 20 to 29, 20 to 30, 21 to 22, 21 to 23, 21 to 24, 21 to 25, 21 to 26, 21 to 27, 21 to 28, 21 to 29, 21 to 30, 22 to 23, 22 to 24, 22 to 25, 22 to 26, 22 to 27, 22 to 28, 22 to 29, 22 to 30, 23 to 24, 23 to 25, 23 to 26, 23 to 27, 23 to 28, 23 to 29, 23 to 30, 24 to 25, 24 to 26, 24 to 27, 24 to 28, 24 to 29, 24 to 30, 25 to 26, 25 to 27, 25 to 28, 25 to 29, 25 to 30, 26 to 27, 26 to 28, 26 to 29, 26 to 30, 27 to 28, 27 to 29, 27 to 30, 28 to 29, 28 to 30, or 29 to 30 linked nucleosides.
D. Certain Modified Oligonucleotides
In certain embodiments, the above modifications (sugar, nucleobase, internucleoside linkage) are incorporated into a modified oligonucleotide. In certain embodiments, modified oligonucleotides are characterized by their modification motifs and overall lengths. In certain embodiments, such parameters are each independent of one another. Thus, unless otherwise indicated, each internucleoside linkage of an oligonucleotide having a gapmer sugar motif may be modified or unmodified and may or may not follow the gapmer modification pattern of the sugar modifications. For example, the internucleoside linkages within the wing regions of a sugar gapmer may be the same or different from one another and may be the same or different from the internucleoside linkages of the gap region of the sugar motif. Likewise, such sugar gapmer oligonucleotides may comprise one or more modified nucleobase independent of the gapmer pattern of the sugar modifications. Unless otherwise indicated, all modifications are independent of nucleobase sequence.
E. Certain Populations of Modified Oligonucleotides
Populations of modified oligonucleotides in which all of the modified oligonucleotides of the population have the same molecular formula can be stereorandom populations or chirally enriched populations. All of the chiral centers of all of the modified oligonucleotides are stereorandom in a stereorandom population. In a chirally enriched population, at least one particular chiral center is not stereorandom in the modified oligonucleotides of the population. In certain embodiments, the modified oligonucleotides of a chirally enriched population are enriched for R-D ribosyl sugar moieties, and all of the phosphorothioate internucleoside linkages are stereorandom. In certain embodiments, the modified oligonucleotides of a chirally enriched population are enriched for both β-D ribosyl sugar moieties and at least one, particular phosphorothioate internucleoside linkage in a particular stereochemical configuration.
F. Nucleobase Sequence
In certain embodiments, oligonucleotides (unmodified or modified oligonucleotides) are further described by their nucleobase sequence. In certain embodiments oligonucleotides have a nucleobase sequence that is complementary to a second oligonucleotide or an identified reference nucleic acid, such as a target nucleic acid. In certain such embodiments, a portion of an oligonucleotide has a nucleobase sequence that is complementary to a second oligonucleotide or an identified reference nucleic acid, such as a target nucleic acid. In certain embodiments, the nucleobase sequence of a portion or entire length of an oligonucleotide is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% complementary to the second oligonucleotide or nucleic acid, such as a target nucleic acid.
II. Certain Oligomeric Compounds
In certain embodiments, provided herein are oligomeric compounds, which consist of an oligonucleotide (modified or unmodified) and optionally one or more conjugate groups and/or terminal groups. Conjugate groups consist of one or more conjugate moiety and a conjugate linker which links the conjugate moiety to the oligonucleotide. Conjugate groups may be attached to either or both ends of an oligonucleotide and/or at any internal position. In certain embodiments, conjugate groups are attached to the 2′-position of a nucleoside of a modified oligonucleotide. In certain embodiments, conjugate groups that are attached to either or both ends of an oligonucleotide are terminal groups. In certain such embodiments, conjugate groups or terminal groups are attached at the 3′ and/or 5′-end of oligonucleotides. In certain such embodiments, conjugate groups (or terminal groups) are attached at the 3′-end of oligonucleotides. In certain embodiments, conjugate groups are attached near the 3′-end of oligonucleotides. In certain embodiments, conjugate groups (or terminal groups) are attached at the 5′-end of oligonucleotides. In certain embodiments, conjugate groups are attached near the 5′-end of oligonucleotides.
Examples of terminal groups include but are not limited to conjugate groups, capping groups, phosphate moieties, protecting groups, modified or unmodified nucleosides, and two or more nucleosides that are independently modified or unmodified.
A. Certain Conjugate Groups
In certain embodiments, oligonucleotides are covalently attached to one or more conjugate groups. In certain embodiments, conjugate groups modify one or more properties of the attached oligonucleotide, including but not limited to pharmacodynamics, pharmacokinetics, stability, binding, absorption, tissue distribution, cellular distribution, cellular uptake, charge and clearance. In certain embodiments, conjugate groups impart a new property on the attached oligonucleotide, e.g., fluorophores or reporter groups that enable detection of the oligonucleotide. Certain conjugate groups and conjugate moieties have been described previously, for example: cholesterol moiety (Letsinger et al., Proc. Natl. Acad. Sci. USA, 1989, 86, 6553-6556), cholic acid (Manoharan et al., Bioorg. Med. Chem. Lett., 1994, 4, 1053-1060), a thioether, e.g., hexyl-S-tritylthiol (Manoharan et al., Ann. N.Y. Acad. Sci., 1992, 660, 306-309; Manoharan et al., Bioorg. Med. Chem. Lett., 1993, 3, 2765-2770), a thiocholesterol (Oberhauser et al., Nucl. Acids Res., 1992, 20, 533-538), an aliphatic chain, e.g., do-decan-diol or undecyl residues (Saison-Behmoaras et al., EMBO J., 1991, 10, 1111-1118; Kabanov et al., FEBS Lett., 1990, 259, 327-330; Svinarchuk et al., Biochimie, 1993, 75, 49-54), a phospholipid, e.g., di-hexadecyl-rac-glycerol or triethyl-ammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate (Manoharan et al., Tetrahedron Lett., 1995, 36, 3651-3654; Shea et al., Nucl. Acids Res., 1990, 18, 3777-3783), a polyamine or a polyethylene glycol chain (Manoharan et al., Nucleosides & Nucleotides, 1995, 14, 969-973), or adamantane acetic acid a palmityl moiety (Mishra et al., Biochim. Biophys. Acta, 1995, 1264, 229-237), an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety (Crooke et al., J. Pharmacol. Exp. Ther., 1996, 277, 923-937), a tocopherol group (Nishina et al., Molecular Therapy Nucleic Acids, 2015, 4, e220; and Nishina et al., Molecular Therapy, 2008, 16, 734-740), or a GalNAc cluster (e.g., WO2014/179620).
1. Conjugate Moieties
Conjugate moieties include, without limitation, intercalators, reporter molecules, polyamines, polyamides, peptides, carbohydrates, vitamin moieties, polyethylene glycols, thioethers, polyethers, cholesterols, thiocholesterols, cholic acid moieties, folate, lipids, phospholipids, biotin, phenazine, phenanthridine, anthraquinone, adamantane, acridine, fluoresceins, rhodamines, coumarins, fluorophores, and dyes.
In certain embodiments, a conjugate moiety comprises an active drug substance, for example, aspirin, warfarin, phenylbutazone, ibuprofen, suprofen, fen-bufen, ketoprofen, (S)-(+)-pranoprofen, carprofen, dansylsarcosine, 2,3,5-triiodobenzoic acid, fingolimod, flufenamic acid, folinic acid, a benzothiadiazide, chlorothiazide, a diazepine, indo-methicin, a barbiturate, a cephalosporin, a sulfa drug, an antidiabetic, an antibacterial or an antibiotic.
2. Conjugate Linkers
Conjugate moieties are attached to oligonucleotides through conjugate linkers. In certain oligomeric compounds, the conjugate linker is a single chemical bond (i.e., the conjugate moiety is attached directly to an oligonucleotide through a single bond). In certain embodiments, the conjugate linker comprises a chain structure, such as a hydrocarbyl chain, or an oligomer of repeating units such as ethylene glycol, nucleosides, or amino acid units.
In certain embodiments, a conjugate linker comprises one or more groups selected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether, thioether, and hydroxylamino. In certain such embodiments, the conjugate linker comprises groups selected from alkyl, amino, oxo, amide and ether groups. In certain embodiments, the conjugate linker comprises groups selected from alkyl and amide groups. In certain embodiments, the conjugate linker comprises groups selected from alkyl and ether groups. In certain embodiments, the conjugate linker comprises at least one phosphorus moiety. In certain embodiments, the conjugate linker comprises at least one phosphate group. In certain embodiments, the conjugate linker includes at least one neutral linking group.
In certain embodiments, conjugate linkers, including the conjugate linkers described above, are bifunctional linking moieties, e.g., those known in the art to be useful for attaching conjugate groups to parent compounds, such as the oligonucleotides provided herein. In general, a bifunctional linking moiety comprises at least two functional groups. One of the functional groups is selected to bind to a particular site on a parent compound and the other is selected to bind to a conjugate group. Examples of functional groups used in a bifunctional linking moiety include but are not limited to electrophiles for reacting with nucleophilic groups and nucleophiles for reacting with electrophilic groups. In certain embodiments, bifunctional linking moieties comprise one or more groups selected from amino, hydroxyl, carboxylic acid, thiol, alkyl, alkenyl, and alkynyl.
Examples of conjugate linkers include but are not limited to pyrrolidine, 8-amino-3,6-dioxaoctanoic acid (ADO), succinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC) and 6-aminohexanoic acid (AHEX or AHA). Other conjugate linkers include but are not limited to substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C2-C10 alkenyl or substituted or unsubstituted C2-C10 alkynyl, wherein a nonlimiting list of preferred substituent groups includes hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl, alkenyl and alkynyl.
In certain embodiments, conjugate linkers comprise 1-10 linker-nucleosides. In certain embodiments, conjugate linkers comprise 2-5 linker-nucleosides. In certain embodiments, conjugate linkers comprise exactly 3 linker-nucleosides. In certain embodiments, conjugate linkers comprise the TCA motif. In certain embodiments, such linker-nucleosides are modified nucleosides. In certain embodiments such linker-nucleosides comprise a modified sugar moiety. In certain embodiments, linker-nucleosides are unmodified. In certain embodiments, linker-nucleosides comprise an optionally protected heterocyclic base selected from a purine, substituted purine, pyrimidine or substituted pyrimidine. In certain embodiments, a cleavable moiety is a nucleoside selected from uracil, thymine, cytosine, 4-N-benzoylcytosine, 5-methyl cytosine, 4-N-benzoyl-5-methyl cytosine, adenine, 6-N-benzoyladenine, guanine and 2-N-isobutyrylguanine. It is typically desirable for linker-nucleosides to be cleaved from the oligomeric compound after it reaches a target tissue. Accordingly, linker-nucleosides are typically linked to one another and to the remainder of the oligomeric compound through cleavable bonds. In certain embodiments, such cleavable bonds are phosphodiester bonds.
Herein, linker-nucleosides are not considered to be part of the oligonucleotide. Accordingly, in embodiments in which an oligomeric compound comprises an oligonucleotide consisting of a specified number or range of linked nucleosides and/or a specified percent complementarity to a reference nucleic acid and the oligomeric compound also comprises a conjugate group comprising a conjugate linker comprising linker-nucleosides, those linker-nucleosides are not counted toward the length of the oligonucleotide and are not used in determining the percent complementarity of the oligonucleotide for the reference nucleic acid. For example, an oligomeric compound may comprise (1) a modified oligonucleotide consisting of 8-30 nucleosides and (2) a conjugate group comprising 1-10 linker-nucleosides that are contiguous with the nucleosides of the modified oligonucleotide. The total number of contiguous linked nucleosides in such an oligomeric compound is more than 30. Alternatively, an oligomeric compound may comprise a modified oligonucleotide consisting of 8-30 nucleosides and no conjugate group. The total number of contiguous linked nucleosides in such an oligomeric compound is no more than 30. Unless otherwise indicated conjugate linkers comprise no more than 10 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 5 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 3 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 2 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 1 linker-nucleoside.
In certain embodiments, it is desirable for a conjugate group to be cleaved from the oligonucleotide. For example, in certain circumstances oligomeric compounds comprising a particular conjugate moiety are better taken up by a particular cell type, but once the oligomeric compound has been taken up, it is desirable that the conjugate group be cleaved to release the unconjugated or parent oligonucleotide. Thus, certain conjugate linkers may comprise one or more cleavable moieties. In certain embodiments, a cleavable moiety is a cleavable bond. In certain embodiments, a cleavable moiety is a group of atoms comprising at least one cleavable bond. In certain embodiments, a cleavable moiety comprises a group of atoms having one, two, three, four, or more than four cleavable bonds. In certain embodiments, a cleavable moiety is selectively cleaved inside a cell or subcellular compartment, such as a lysosome. In certain embodiments, a cleavable moiety is selectively cleaved by endogenous enzymes, such as nucleases.
In certain embodiments, a cleavable bond is selected from among an amide, an ester, an ether, one or both esters of a phosphodiester, a phosphate ester, a carbamate, and a disulfide. In certain embodiments, a cleavable bond is one or both of the esters of a phosphodiester. In certain embodiments, a cleavable moiety comprises a phosphate or phosphodiester. In certain embodiments, the cleavable moiety is a phosphate or phosphodiester linkage between an oligonucleotide and a conjugate moiety or conjugate group.
In certain embodiments, a cleavable moiety comprises or consists of one or more linker-nucleosides. In certain such embodiments, the one or more linker-nucleosides are linked to one another and/or to the remainder of the oligomeric compound through cleavable bonds. In certain embodiments, such cleavable bonds are unmodified phosphodiester bonds. In certain embodiments, a cleavable moiety is 2′-deoxynucleoside that is attached to either the 3′ or 5′-terminal nucleoside of an oligonucleotide by a phosphodiester internucleoside linkage and covalently attached to the remainder of the conjugate linker or conjugate moiety by a phosphate or phosphorothioate linkage. In certain such embodiments, the cleavable moiety is 2′-deoxyadenosine.
3. Cell-Targeting Moieties
In certain embodiments, a conjugate group comprises a cell-targeting moiety. In certain embodiments, a conjugate group has the general formula:
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wherein n is from 1 to about 3, m is 0 when n is 1, m is 1 when n is 2 or greater, j is 1 or 0, and k is 1 or 0.
In certain embodiments, n is 1, j is 1 and k is 0. In certain embodiments, n is 1, j is 0 and k is 1. In certain embodiments, n is 1, j is 1 and k is 1. In certain embodiments, n is 2, j is 1 and k is 0. In certain embodiments, n is 2, j is 0 and k is 1. In certain embodiments, n is 2, j is 1 and k is 1. In certain embodiments, n is 3, j is 1 and k is 0. In certain embodiments, n is 3, j is 0 and k is 1. In certain embodiments, n is 3, j is 1 and k is 1.
In certain embodiments, conjugate groups comprise cell-targeting moieties that have at least one tethered ligand. In certain embodiments, cell-targeting moieties comprise two tethered ligands covalently attached to a branching group. In certain embodiments, cell-targeting moieties comprise three tethered ligands covalently attached to a branching group.
In certain embodiments, the cell-targeting moiety comprises a branching group comprising one or more groups selected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether, thioether and hydroxylamino groups. In certain embodiments, the branching group comprises a branched aliphatic group comprising groups selected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether, thioether and hydroxylamino groups. In certain such embodiments, the branched aliphatic group comprises groups selected from alkyl, amino, oxo, amide and ether groups. In certain such embodiments, the branched aliphatic group comprises groups selected from alkyl, amino and ether groups. In certain such embodiments, the branched aliphatic group comprises groups selected from alkyl and ether groups. In certain embodiments, the branching group comprises a mono or polycyclic ring system.
In certain embodiments, each tether of a cell-targeting moiety comprises one or more groups selected from alkyl, substituted alkyl, ether, thioether, disulfide, amino, oxo, amide, phosphodiester, and polyethylene glycol, in any combination. In certain embodiments, each tether is a linear aliphatic group comprising one or more groups selected from alkyl, ether, thioether, disulfide, amino, oxo, amide, and polyethylene glycol, in any combination. In certain embodiments, each tether is a linear aliphatic group comprising one or more groups selected from alkyl, phosphodiester, ether, amino, oxo, and amide, in any combination. In certain embodiments, each tether is a linear aliphatic group comprising one or more groups selected from alkyl, ether, amino, oxo, and amid, in any combination. In certain embodiments, each tether is a linear aliphatic group comprising one or more groups selected from alkyl, amino, and oxo, in any combination. In certain embodiments, each tether is a linear aliphatic group comprising one or more groups selected from alkyl and oxo, in any combination. In certain embodiments, each tether is a linear aliphatic group comprising one or more groups selected from alkyl and phosphodiester, in any combination. In certain embodiments, each tether comprises at least one phosphorus linking group or neutral linking group. In certain embodiments, each tether comprises a chain from about 6 to about 20 atoms in length. In certain embodiments, each tether comprises a chain from about 10 to about 18 atoms in length. In certain embodiments, each tether comprises about 10 atoms in chain length.
In certain embodiments, each ligand of a cell-targeting moiety has an affinity for at least one type of receptor on a target cell. In certain embodiments, each ligand has an affinity for at least one type of receptor on the surface of a mammalian liver cell. In certain embodiments, each ligand has an affinity for the hepatic asialoglycoprotein receptor (ASGP-R). In certain embodiments, each ligand is a carbohydrate. In certain embodiments, each ligand is, independently selected from galactose, N-acetyl galactosamine (GalNAc), mannose, glucose, glucosamine and fucose. In certain embodiments, each ligand is N-acetyl galactosamine (GalNAc). In certain embodiments, the cell-targeting moiety comprises 3 GalNAc ligands. In certain embodiments, the cell-targeting moiety comprises 2 GalNAc ligands. In certain embodiments, the cell-targeting moiety comprises 1 GalNAc ligand.
In certain embodiments, each ligand of a cell-targeting moiety is a carbohydrate, carbohydrate derivative, modified carbohydrate, polysaccharide, modified polysaccharide, or polysaccharide derivative. In certain such embodiments, the conjugate group comprises a carbohydrate cluster (see, e.g., Maier et al., “Synthesis of Antisense Oligonucleotides Conjugated to a Multivalent Carbohydrate Cluster for Cellular Targeting,” Bioconjugate Chemistry, 2003, 14, 18-29, or Rensen et al., “Design and Synthesis of Novel N-Acetylgalactosamine-Terminated Glycolipids for Targeting of Lipoproteins to the Hepatic Asiaglycoprotein Receptor,” J. Med. Chem. 2004, 47, 5798-5808, which are incorporated herein by reference in their entirety). In certain such embodiments, each ligand is an amino sugar or a thio sugar. For example, amino sugars may be selected from any number of compounds known in the art, such as sialic acid, α-D-galactosamine, β-muramic acid, 2-deoxy-2-methylamino-L-glucopyranose, 4,6-dideoxy-4-formamido-2,3-di-O-methyl-D-mannopyranose, 2-deoxy-2-sulfoamino-D-glucopyranose and N-sulfo-D-glucosamine, and N-glycoloyl-α-neuraminic acid. For example, thio sugars may be selected from 5-Thio-β-D-glucopyranose, methyl 2,3,4-tri-O-acetyl-1-thio-6-O-trityl-α-D-glucopyranoside, 4-thio-β-D-galactopyranose, and ethyl 3,4,6,7-tetra-O-acetyl-2-deoxy-1,5-dithio-α-D-gluco-heptopyranoside.
In certain embodiments, conjugate groups comprise a cell-targeting moiety having the formula:
![embedded image]()
In certain embodiments, conjugate groups comprise a cell-targeting moiety having the formula:
![embedded image]()
In certain embodiments, conjugate groups comprise a cell-targeting moiety having the formula:
![embedded image]()
In certain embodiments, compounds described herein comprise a conjugate group described herein as “THA-GalNAac3”. THA-GalNAc3 is shown below without the optional cleavable moiety at the end of the linker region:
![embedded image]()
In certain embodiments, compounds described herein comprise THA-GalNAc3-phosphate, also represented as (THA-GalNAc3)o, having the formula:
![embedded image]()
wherein modified oligonucleotide represents a modified oligonucleotide.
In certain embodiments, compounds described herein comprise a 3′-THA-C6-GalNAc hydroxyproline phosphate, (“HPPO-GalNAc”). 3′-HPPO-GalNAc is represented by the structure below wherein the phosphate group is attached to the 3′-oxygen atom of the 3′ nucleoside:
![embedded image]()
Representative publications that teach the preparation of certain of the above noted conjugate groups and compounds comprising conjugate groups, tethers, conjugate linkers, branching groups, ligands, cleavable moieties as well as other modifications include without limitation, U.S. Pat. Nos. 5,994,517, 6,300,319, 6,660,720, 6,906,182, 7,262,177, 7,491,805, 8,106,022, 7,723,509, 9,127,276, US 2006/0148740, US 2011/0123520, WO 2013/033230 and WO 2012/037254, Biessen et al., J. Med. Chem. 1995, 38, 1846-1852, Lee et al., Bioorganic & Medicinal Chemistry 2011, 19, 2494-2500, Rensen et al., J. Biol. Chem. 2001, 276, 37577-37584, Rensen et al., J. Med. Chem. 2004, 47, 5798-5808, Sliedregt et al., J. Med. Chem. 1999, 42, 609-618, and Valentijn et al., Tetrahedron, 1997, 53, 759-770, each of which is incorporated by reference herein in its entirety.
In certain embodiments, compounds described herein comprise modified oligonucleotides comprising a gapmer or fully modified motif and a conjugate group comprising at least one, two, or three GalNAc ligands. In certain embodiments compounds described herein comprise a conjugate group found in any of the following references: Lee, Carbohydr Res, 1978, 67, 509-514; Connolly et al., J Biol Chem, 1982, 257, 939-945; Pavia et al., Int J Pep Protein Res, 1983, 22, 539-548; Lee et al., Biochem, 1984, 23, 4255-4261; Lee et al., Glycoconjugate J, 1987, 4, 317-328; Toyokuni et al., Tetrahedron Lett, 1990, 31, 2673-2676; Biessen et al., J Med Chem, 1995, 38, 1538-1546; Valentijn et al., Tetrahedron, 1997, 53, 759-770; Kim et al., Tetrahedron Lett, 1997, 38, 3487-3490; Lee et al., Bioconjug Chem, 1997, 8, 762-765; Kato et al., Glycobiol, 2001, 11, 821-829; Rensen et al., J Biol Chem, 2001, 276, 37577-37584; Lee et al., Methods Enzymol, 2003, 362, 38-43; Westerlind et al., Glycoconj J, 2004, 21, 227-241; Lee et al., Bioorg Med Chem Lett, 2006, 16(19), 5132-5135; Maierhofer et al., Bioorg Med Chem, 2007, 15, 7661-7676; Khorev et al., Bioorg Med Chem, 2008, 16, 5216-5231; Lee et al., Bioorg Med Chem, 2011, 19, 2494-2500; Kornilova et al., Analyt Biochem, 2012, 425, 43-46; Pujol et al., Angew Chemie Int Ed Engl, 2012, 51, 7445-7448; Biessen et al., J Med Chem, 1995, 38, 1846-1852; Sliedregt et al., J Med Chem, 1999, 42, 609-618; Rensen et al., J Med Chem, 2004, 47, 5798-5808; Rensen et al., Arterioscler Thromb Vasc Biol, 2006, 26, 169-175; van Rossenberg et al., Gene Ther, 2004, 11, 457-464; Sato et al., J Am Chem Soc, 2004, 126, 14013-14022; Lee et al., J Org Chem, 2012, 77, 7564-7571; Biessen et al., FASEB J, 2000, 14, 1784-1792; Rajur et al., Bioconjug Chem, 1997, 8, 935-940; Duff et al., Methods Enzymol, 2000, 313, 297-321; Maier et al., Bioconjug Chem, 2003, 14, 18-29; Jayaprakash et al., Org Lett, 2010, 12, 5410-5413; Manoharan, Antisense Nucleic Acid Drug Dev, 2002, 12, 103-128; Merwin et al., Bioconjug Chem, 1994, 5, 612-620; Tomiya et al., Bioorg Med Chem, 2013, 21, 5275-5281; International applications WO1998/013381; WO2011/038356; WO1997/046098; WO2008/098788; WO2004/101619; WO2012/037254; WO2011/120053; WO2011/100131; WO2011/163121; WO2012/177947; WO2013/033230; WO2013/075035; WO2012/083185; WO2012/083046; WO2009/082607; WO2009/134487; WO2010/144740; WO2010/148013; WO1997/020563; WO2010/088537; WO2002/043771; WO2010/129709; WO2012/068187; WO2009/126933; WO2004/024757; WO2010/054406; WO2012/089352; WO2012/089602; WO2013/166121; WO2013/165816; U.S. Pat. Nos. 4,751,219; 8,552,163; 6,908,903; 7,262,177; 5,994,517; 6,300,319; 8,106,022; 7,491,805; 7,491,805; 7,582,744; 8,137,695; 6,383,812; 6,525,031; 6,660,720; 7,723,509; 8,541,548; 8,344,125; 8,313,772; 8,349,308; 8,450,467; 8,501,930; 8,158,601; 7,262,177; 6,906,182; 6,620,916; 8,435,491; 8,404,862; 7,851,615; Published U.S. Patent Application Publications US2011/0097264; US2011/0097265; US2013/0004427; US2005/0164235; US2006/0148740; US2008/0281044; US2010/0240730; US2003/0119724; US2006/0183886; US2008/0206869; US2011/0269814; US2009/0286973; US2011/0207799; US2012/0136042; US2012/0165393; US2008/0281041; US2009/0203135; US2012/0035115; US2012/0095075; US2012/0101148; US2012/0128760; US2012/0157509; US2012/0230938; US2013/0109817; US2013/0121954; US2013/0178512; US2013/0236968; US2011/0123520; US2003/0077829; US2008/0108801; and US2009/0203132; each of which is incorporated by reference in its entirety.
B. Certain Terminal Groups
In certain embodiments, oligomeric compounds comprise one or more terminal groups. In certain such embodiments, oligomeric compounds comprise a stabilized 5′-phosphate. Stabilized 5′-phosphates include, but are not limited to 5′-phosphinates, including, but not limited to 5′-vinylphosphonates. In certain embodiments, terminal groups comprise one or more abasic nucleosides and/or inverted nucleosides. In certain embodiments, terminal groups comprise one or more 2′-linked nucleosides. In certain such embodiments, the 2′-linked nucleoside is an abasic nucleoside.
III. Oligomeric Duplexes
In certain embodiments, oligomeric compounds described herein comprise an oligonucleotide, having a nucleobase sequence complementary to that of a target nucleic acid. In certain embodiments, an oligomeric compound is paired with a second oligomeric compound to form an oligomeric duplex. Such oligomeric duplexes comprise a first oligomeric compound having a portion complementary to a target nucleic acid and a second oligomeric compound having a portion complementary to the first oligomeric compound. In certain embodiments, the first oligomeric compound of an oligomeric duplex comprises or consists of (1) a modified or unmodified oligonucleotide and optionally a conjugate group and (2) a second modified or unmodified oligonucleotide and optionally a conjugate group. Either or both oligomeric compounds of an oligomeric duplex may comprise a conjugate group. The oligonucleotides of each oligomeric compound of an oligomeric duplex may include non-complementary overhanging nucleosides.
IV. Antisense Activity
In certain embodiments, oligomeric compounds and oligomeric duplexes are capable of hybridizing to a target nucleic acid, resulting in at least one antisense activity; such oligomeric compounds and oligomeric duplexes are antisense compounds. In certain embodiments, antisense compounds have antisense activity when they reduce or inhibit the amount or activity of a target nucleic acid by 25% or more in the standard cell assay. In certain embodiments, antisense compounds selectively affect one or more target nucleic acid. Such antisense compounds comprise a nucleobase sequence that hybridizes to one or more target nucleic acid, resulting in one or more desired antisense activity and does not hybridize to one or more non-target nucleic acid or does not hybridize to one or more non-target nucleic acid in such a way that results in significant undesired antisense activity.
In certain antisense activities, hybridization of an antisense compound to a target nucleic acid results in recruitment of a protein that cleaves the target nucleic acid. For example, certain antisense compounds result in RNase H mediated cleavage of the target nucleic acid. RNase H is a cellular endonuclease that cleaves the RNA strand of an RNA:DNA duplex. The DNA in such an RNA:DNA duplex need not be unmodified DNA. In certain embodiments, described herein are antisense compounds that are sufficiently “DNA-like” to elicit RNase H activity. In certain embodiments, one or more non-DNA-like nucleoside in the gap of a gapmer is tolerated.
In certain antisense activities, an antisense compound or a portion of an antisense compound is loaded into an RNA-induced silencing complex (RISC), ultimately resulting in cleavage of the target nucleic acid. For example, certain antisense compounds result in cleavage of the target nucleic acid by Argonaute. Antisense compounds that are loaded into RISC are RNAi compounds. RNAi compounds may be double-stranded (siRNA) or single-stranded (ssRNA).
In certain embodiments, hybridization of an antisense compound to a target nucleic acid does not result in recruitment of a protein that cleaves that target nucleic acid. In certain embodiments, hybridization of the antisense compound to the target nucleic acid results in alteration of splicing of the target nucleic acid. In certain embodiments, hybridization of an antisense compound to a target nucleic acid results in inhibition of a binding interaction between the target nucleic acid and a protein or other nucleic acid. In certain embodiments, hybridization of an antisense compound to a target nucleic acid results in alteration of translation of the target nucleic acid.
Antisense activities may be observed directly or indirectly. In certain embodiments, observation or detection of an antisense activity involves observation or detection of a change in an amount of a target nucleic acid or protein encoded by such target nucleic acid, a change in the ratio of splice variants of a nucleic acid or protein and/or a phenotypic change in a cell or subject.
V. Certain Target Nucleic Acids
In certain embodiments, oligomeric compounds comprise or consist of an oligonucleotide comprising a portion that is complementary to a target nucleic acid. In certain embodiments, the target nucleic acid is an endogenous RNA molecule. In certain embodiments, the target nucleic acid encodes a protein. In certain such embodiments, the target nucleic acid is selected from: a mature mRNA, and a pre-mRNA, including intronic, exonic and untranslated regions. In certain embodiments, the target RNA is an antisense transcript. In certain embodiments, the target RNA is a mature mRNA. In certain embodiments, the target nucleic acid is a pre-mRNA. In certain such embodiments, the target region is entirely within an intron. In certain embodiments, the target region spans an intron/exon junction. In certain embodiments, the target region is at least 50% within an intron. In certain embodiments, the target nucleic acid is the RNA transcriptional product of a retrogene. In certain embodiments, the target nucleic acid is a non-coding RNA. In certain such embodiments, the target non-coding RNA is selected from: a long non-coding RNA, a short non-coding RNA, an intronic RNA molecule.
A. Complementarity/Mismatches to the Target Nucleic Acid
It is possible to introduce mismatch bases without eliminating activity. For example, Gautschi et al (J. Natl. Cancer Inst. 93:463-471, March 2001) demonstrated the ability of an oligonucleotide having 100% complementarity to the bcl-2 mRNA and having 3 mismatches to the bcl-xL mRNA to reduce the expression of both bcl-2 and bcl-xL in vitro and in vivo. Furthermore, this oligonucleotide demonstrated potent anti-tumor activity in vivo. Maher and Dolnick (Nuc. Acid. Res. 16:3341-3358, 1988) tested a series of tandem 14 nucleobase oligonucleotides, and 28 and 42 nucleobase oligonucleotides comprised of the sequence of two or three of the tandem oligonucleotides, respectively, for their ability to arrest translation of human DHFR in a rabbit reticulocyte assay. Each of the three 14 nucleobase oligonucleotides alone was able to inhibit translation, albeit at a more modest level than the 28 or 42 nucleobase oligonucleotides.
In certain embodiments, oligonucleotides are complementary to the target nucleic acid over the entire length of the oligonucleotide. In certain embodiments, oligonucleotides are 99%, 95%, 90%, 85%, or 80% complementary to the target nucleic acid. In certain embodiments, oligonucleotides are at least 80% complementary to the target nucleic acid over the entire length of the oligonucleotide and comprise a portion that is 100% or fully complementary to a target nucleic acid. In certain embodiments, the portion of full complementarity is 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 nucleobases in length.
In certain embodiments, oligonucleotides comprise one or more mismatched nucleobases relative to the target nucleic acid. In certain embodiments, antisense activity against the target is reduced by such mismatch, but activity against a non-target is reduced by a greater amount. Thus, in certain embodiments selectivity of the oligonucleotide is improved. In certain embodiments, the mismatch is specifically positioned within an oligonucleotide having a gapmer motif. In certain embodiments, the mismatch is at position 1, 2, 3, 4, 5, 6, 7, or 8 from the 5′-end of the gap region. In certain embodiments, the mismatch is at position 9, 8, 7, 6, 5, 4, 3, 2, 1 from the 3′-end of the gap region. In certain embodiments, the mismatch is at position 1, 2, 3, or 4 from the 5′-end of the wing region. In certain embodiments, the mismatch is at position 4, 3, 2, or 1 from the 3′-end of the wing region.
B. FXII
In certain embodiments, oligomeric compounds comprise or consist of an oligonucleotide, or portion thereof, that is complementary to a target nucleic acid, wherein the target nucleic acid is a FXII nucleic acid. In certain embodiments, the FXII nucleic acid has the human FXII mRNA sequence designated herein as SEQ ID NO: 1 (ENSEMBL ID ENST00000253496.3 from ENSEMBL version 99: January 2020). In certain embodiments, the FXII nucleic acid has the human F12 genomic sequence, designated herein as SEQ ID NO: 2 (ENSEMBL ID ENSG00000131187.9 from ENSEML version 99: January 2020, human reference assembly version GRCh38.p13 located on the reverse strand of chromosome 5 from positions 177,402,140 to 177,409,576). In certain embodiments, the FXII nucleic acid has the human F12 genomic sequence, designated herein as SEQ ID NO. 3 (the complement of GENBANK Accession No. NC_000005.10 truncated from truncated from nucleotides 177399001 to 177413000). In certain embodiments, the FXII nucleic acid has the human FXII mRNA sequence designated herein as SEQ ID NO: 4 (GENBANK Accession No. NM_000505.3).
In certain embodiments, an oligomeric compound complementary to any one of SEQ ID NOS: 1-4 is capable of reducing FXII RNA in a cell. In certain embodiments, an oligomeric compound complementary to any one of SEQ ID NOS: 1-4 is capable of reducing FXII protein in a cell. In certain embodiments, the cell is in vitro. In certain embodiments, the cell is in a subject. In certain embodiments, an oligomeric compound complementary to any one of SEQ ID NOS: 1-4 is capable of ameliorating one or more symptoms of a thromboembolic condition when administered to a subject. In certain embodiments, the thromboembolic condition is deep vein thrombosis, venous thrombosis, arterial thrombosis, pulmonary embolism, myocardial infarction, or stroke. In certain embodiments, the symptoms are selected from pain, shortness of breath, heart burn, cold sweat, fatigue, lightheadedness, dizziness, swelling, cramping, and death.
In certain embodiments, an oligomeric compound complementary to any one of SEQ ID NOS: 1-4 is capable of reducing the detectable amount of FXII RNA in vitro by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%. In certain embodiments, an oligomeric compound complementary to any one of SEQ ID NOS: 1-4 is capable of reducing the detectable amount of FXII protein in vitro by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%. In certain embodiments, an oligomeric compound complementary to any one of SEQ ID NOS: 1-4 is capable of reducing the detectable amount of FXII RNA in a biological sample of a subject by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%. In certain embodiments, an oligomeric compound complementary to any one of SEQ ID NOS: 1-4 is capable of reducing the detectable amount of FXII protein in a biological sample of a subject by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%. In certain embodiments, the biological sample comprises blood, plasma/serum, or a cell of the subject.
VI. Certain Pharmaceutical Compositions
In certain embodiments, described herein are pharmaceutical compositions comprising one or more oligomeric compounds. In certain embodiments, the one or more oligomeric compounds each consists of a modified oligonucleotide. In certain embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable diluent or carrier. In certain embodiments, a pharmaceutical composition comprises or consists of a sterile saline solution and one or more oligomeric compound. In certain embodiments, the sterile saline is pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition comprises or consists of one or more oligomeric compound and sterile water. In certain embodiments, the sterile water is pharmaceutical grade water. In certain embodiments, a pharmaceutical composition comprises or consists of one or more oligomeric compound and phosphate-buffered saline (PBS). In certain embodiments, the sterile PBS is pharmaceutical grade PBS. In certain embodiments, a pharmaceutical composition comprises or consists of one or more oligomeric compound and artificial cerebrospinal fluid. In certain embodiments, the artificial cerebrospinal fluid is pharmaceutical grade.
In certain embodiments, a pharmaceutical composition comprises a modified oligonucleotide and artificial cerebrospinal fluid. In certain embodiments, a pharmaceutical composition consists of a modified oligonucleotide and artificial cerebrospinal fluid. In certain embodiments, a pharmaceutical composition consists essentially of a modified oligonucleotide and artificial cerebrospinal fluid. In certain embodiments, the artificial cerebrospinal fluid is pharmaceutical grade.
In certain embodiments, pharmaceutical compositions comprise one or more oligomeric compound and one or more excipients. In certain embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose and polyvinylpyrrolidone.
In certain embodiments, oligomeric compounds may be admixed with pharmaceutically acceptable active and/or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.
In certain embodiments, pharmaceutical compositions comprising an oligomeric compound encompass any pharmaceutically acceptable salts of the oligomeric compound, esters of the oligomeric compound, or salts of such esters. In certain embodiments, pharmaceutical compositions comprising oligomeric compounds comprising one or more oligonucleotide, upon administration to a subject, including a human, are capable of providing (directly or indirectly) the biologically active metabolite or residue thereof. Accordingly, for example, the disclosure is also drawn to pharmaceutically acceptable salts of oligomeric compounds, prodrugs, pharmaceutically acceptable salts of such prodrugs, and other bioequivalents. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts. In certain embodiments, prodrugs comprise one or more conjugate group attached to an oligonucleotide, wherein the conjugate group is cleaved by endogenous nucleases within the body.
Lipid moieties have been used in nucleic acid therapies in a variety of methods. In certain such methods, the nucleic acid, such as an oligomeric compound, is introduced into preformed liposomes or lipoplexes made of mixtures of cationic lipids and neutral lipids. In certain methods, DNA complexes with mono- or poly-cationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of a pharmaceutical agent to a particular cell or tissue. In certain embodiments, a lipid moiety is selected to increase distribution of a pharmaceutical agent to fat tissue. In certain embodiments, a lipid moiety is selected to increase distribution of a pharmaceutical agent to muscle tissue.
In certain embodiments, pharmaceutical compositions comprise a delivery system. Examples of delivery systems include, but are not limited to, liposomes and emulsions. Certain delivery systems are useful for preparing certain pharmaceutical compositions including those comprising hydrophobic compounds. In certain embodiments, certain organic solvents such as dimethylsulfoxide are used.
In certain embodiments, pharmaceutical compositions comprise one or more tissue-specific delivery molecules designed to deliver the one or more pharmaceutical agents of the present invention to specific tissues or cell types. For example, in certain embodiments, pharmaceutical compositions include liposomes coated with a tissue-specific antibody.
In certain embodiments, pharmaceutical compositions comprise a co-solvent system. Certain of such co-solvent systems comprise, for example, benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. In certain embodiments, such co-solvent systems are used for hydrophobic compounds. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80™ and 65% w/v polyethylene glycol 300. The proportions of such co-solvent systems may be varied considerably without significantly altering their solubility and toxicity characteristics. Furthermore, the identity of co-solvent components may be varied: for example, other surfactants may be used instead of Polysorbate 80™; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides may substitute for dextrose.
In certain embodiments, pharmaceutical compositions are prepared for oral administration. In certain embodiments, pharmaceutical compositions are prepared for buccal administration. In certain embodiments, a pharmaceutical composition is prepared for administration by injection (e.g., intravenous, subcutaneous, intramuscular, intrathecal (IT), intracerebroventricular (ICV), etc.). In certain of such embodiments, a pharmaceutical composition comprises a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. In certain embodiments, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives). In certain embodiments, injectable suspensions are prepared using appropriate liquid carriers, suspending agents and the like. Certain pharmaceutical compositions for injection are presented in unit dosage form, e.g., in ampoules or in multi-dose containers. Certain pharmaceutical compositions for injection are suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Certain solvents suitable for use in pharmaceutical compositions for injection include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes.
Under certain conditions, certain compounds disclosed herein act as acids. Although such compounds may be drawn or described in protonated (free acid) form, or ionized and in association with a cation (salt) form, aqueous solutions of such compounds exist in equilibrium among such forms. For example, a phosphate linkage of an oligonucleotide in aqueous solution exists in equilibrium among free acid, anion and salt forms. Unless otherwise indicated, compounds described herein are intended to include all such forms. Moreover, certain oligonucleotides have several such linkages, each of which is in equilibrium. Thus, oligonucleotides in solution exist in an ensemble of forms at multiple positions all at equilibrium. The term “oligonucleotide” is intended to include all such forms. Drawn structures necessarily depict a single form. Nevertheless, unless otherwise indicated, such drawings are likewise intended to include corresponding forms. Herein, a structure depicting the free acid of a compound followed by the term “or a salt thereof” expressly includes all such forms that may be fully or partially protonated/de-protonated/in association with a cation. In certain instances, one or more specific cation is identified.
In certain embodiments, modified oligonucleotides or oligomeric compounds are in aqueous solution with sodium. In certain embodiments, modified oligonucleotides or oligomeric compounds are in aqueous solution with potassium. In certain embodiments, modified oligonucleotides or oligomeric compounds are in PBS. In certain embodiments, modified oligonucleotides or oligomeric compounds are in water. In certain such embodiments, the pH of the solution is adjusted with NaOH and/or HCl to achieve a desired pH.
VII. Certain Compositions
1. Compound No. 1194357
In certain embodiments, Compound No. 1194357 is characterized as an oligomeric compound consisting of a conjugate group and a modified oligonucleotide, wherein the conjugate group is a THA-GalNAc3 that is directly attached to the 5′ end of the modified oligonucleotide through a phosphodiester linkage, (THA-GalNAc3)o; wherein (THA-GalNAc3)o is represented by the following structure:
![embedded image]()
wherein the modified oligonucleotide is a 3-10-3 cEt gapmer, having a sequence of (from 5′ to 3′) GCGGAATCACCAAGGA (incorporated herein as SEQ ID NO: 3379); wherein each of nucleosides 1-3 and 14-16 (from 5′ to 3′) comprise a cEt modified sugar moiety and each of nucleosides 4-13 are 2′-β-D-deoxynucleosides; wherein each internucleoside linkage is a phosphorothioate internucleoside linkage; and wherein each cytosine is a 5-methyl cytosine.
In certain embodiments, Compound No. 1194357 is represented by the following chemical notation: (THA-GalNAc3)o Gks mCks Gks Gds Ads Ads Tds mCds Ads mCds mCds Ads Ads Gks Gks Ak, wherein:
- A=an adenine nucleobase,
- mC=a 5-methyl cytosine nucleobase,
- G=a guanine nucleobase,
- T=a thymine nucleobase,
- k=a cEt modified sugar moiety,
- d=a 2′-β-D-deoxyribosyl sugar moiety, and
- s=a phosphorothioate internucleoside linkage.
In certain embodiments, provided is an oligomeric compound comprising a modified oligonucleotide and a conjugate group according to the following chemical notation: (THA-GalNAc3)o Gks mCks Gks Gds Ads Ads Tds mCds Ads mCds mCds Ads Ads Gks Gks Ak, wherein:
- A=an adenine nucleobase,
- mC=a 5-methyl cytosine nucleobase,
- G=a guanine nucleobase,
- T=a thymine nucleobase,
- k=a cEt modified sugar moiety,
- d=a 2′-β-D-deoxyribosyl sugar moiety, and
- s=a phosphorothioate internucleoside linkage;
and wherein (THA-GalNAc3)o=
![embedded image]()
In certain embodiments, Compound No. 1194357 is represented by the following chemical structure:
![embedded image]()
(SEQ ID NO: 3379). Structure 1. Compound No. 1194357
In certain embodiments, the sodium salt of Compound No. 1194357 is represented by the following chemical structure:
![embedded image]()
(SEQ ID NO: 3379). Structure 2. The sodium salt of Compound No. 1194357
2. Compound No. 1270705
In certain embodiments, Compound No. 1270705 is characterized as an oligomeric compound consisting of a conjugate group and a modified oligonucleotide; wherein the conjugate group is a THA-GalNAc3 that is directly attached to the 5′ end of the modified oligonucleotide through a phosphodiester linkage, (THA-GalNAc3)o; wherein (THA-GalNAc3)o is represented by the following structure:
![embedded image]()
wherein the modified oligonucleotide is gapmer, having a sequence of (from 5′ to 3′) GCACUTTATTGAGTTC (incorporated herein as SEQ ID NO: 5006); wherein each of nucleosides 1-3 and 14-16 (from 5′ to 3′) comprise a cEt modified sugar moiety, each of nucleosides 4 and 6-13 are 2′-β-D-deoxynucleosides, and nucleoside 5 comprises a 2′-O-methyl sugar moiety; wherein each internucleoside linkage is a phosphorothioate internucleoside linkage, and wherein each cytosine is a 5-methyl cytosine.
In certain embodiments, Compound No. 1270705 is represented by the following chemical notation: (THA-GalNAc3)o Gks mCks Aks mCds Uys Tds Tds Ads Tds Tds Gds Ads Gds Tks Tks mCk, wherein:
- A=an adenine nucleobase,
- mC=a 5-methyl cytosine nucleobase,
- G=a guanine nucleobase,
- T=a thymine nucleobase,
- U=a uracil nucleobase,
- k=a cEt modified sugar moiety,
- y=a 2′-O-methyl modified ribosyl sugar moiety,
- d=a 2′-β-D-deoxyribosyl sugar moiety, and
- s=a phosphorothioate internucleoside linkage.
In certain embodiments, provided is an oligomeric compound comprising a modified oligonucleotide and a conjugate group according to the following chemical notation: (THA-GalNAc3)o Gks mCks Aks mCds Uys Tds Tds Ads Tds Tds Gds Ads Gds Tks Tks mCk, wherein:
- A=an adenine nucleobase,
- mC=a 5-methyl cytosine nucleobase,
- G=a guanine nucleobase,
- T=a thymine nucleobase,
- U=a uracil nucleobase,
- k=a cEt modified sugar moiety,
- y=a 2′-O-methyl modified ribosyl sugar moiety,
- d=a 2′-β-D-deoxyribosyl sugar moiety, and
- s=a phosphorothioate internucleoside linkage;
and wherein (THA-GalNAc3)o=
![embedded image]()
In certain embodiments, Compound No. 1270705 is represented by the following chemical structure:
![embedded image]()
![embedded image]()
(SEQ ID NO: 5006).
Structure 3. Compound No. 1270705
In certain embodiments, the sodium salt of Compound No. 1270705 is represented by the following chemical structure:
![embedded image]()
![embedded image]()
(SEQ ID NO: 5006).
Structure 4. The sodium salt of Compound No. 1270705
VIII. Certain Comparator Compounds
Comparator Compound Nos. 1213029, 1213030, 1213034, and 1358035 (relating to Compound Nos. A-145667, A-145668, A-145676, and A-145669, respectively) were selected as comparator compounds from Example 1 of WO2017/120397 (incorporated herein by reference).
In certain embodiments, Compound No. 1213029, a surrogate of which (Compound A-145667) is provided in WO2017/120397, was used as a comparator compound, see Example 14. Compound No. 1213029 is a gapmer having a sugar motif of (from 5′ to 3′) yyyyyddddddddddyyyyy, wherein each “y” represents a 2′-O-methyl sugar moiety and each “d” represents a 2′-β-D-deoxyribosyl sugar moiety; having a sequence of (from 5′ to 3′) GAAUACCAAGGAGGGAAAG (SEQ ID NO: 5042), wherein each “C” of the gap is a 5-methyl cytosine; and wherein each internucleoside linkage is a phosphorothioate internucleoside linkage.
In certain embodiments, Compound No. 1213030, a surrogate of which (Compound A-145668) is provided in WO2017/120397, was used as a comparator compound, see Example 14. Compound No. 1213030 is a gapmer having a sugar motif of (from 5′ to 3′) yyyyyddddddddddyyyyy, wherein each “y” represents a 2′-O-methyl sugar moiety and each “d” represents a 2′-β-D-deoxyribosyl sugar moiety; having a sequence of (from 5′ to 3′) UCUCACTGCGGAATCACCAA (SEQ ID NO: 5041), wherein each “C” of the gap is a 5-methyl cytosine; and wherein each internucleoside linkage is a phosphorothioate internucleoside linkage.
In certain embodiments, Compound No. 1213034, a surrogate of which (Compound A-145676) is provided in WO2017/120397, was used as a comparator compound, see Example 14. Compound No. 1213034 is a gapmer having a sugar motif of (from 5′ to 3′) yyyyyddddddddddyyyyy, wherein each “y” represents a 2′-O-methyl sugar moiety and each “d” represents a 2′-β-D-deoxyribosyl sugar moiety; having a sequence of (from 5′ to 3′) CACUUTATTGAGTTCCUGCG (SEQ ID NO: 5039), wherein each “C” of the gap is a 5-methyl cytosine; and wherein each internucleoside linkage is a phosphorothioate internucleoside linkage.
In certain embodiments, Compound No. 1358035, a surrogate of which (Compound A-145669) is provided in WO2017/120397, was used as a comparator compound, see Example 14. Compound No. 1358035 is a gapmer having a sugar motif of (from 5′ to 3′) yyyyyddddddddddyyyyy, wherein each “y” represents a 2′-O-methyl sugar moiety and each “d” represents a 2′-β-D-deoxyribosyl sugar moiety; having a sequence of (from 5′ to 3′) CCCCAGCCACTCTCTCACUG (SEQ ID NO: 5040), wherein each “C” of the gap is a 5-methyl cytosine; and wherein each internucleoside linkage is a phosphorothioate internucleoside linkage.
In certain embodiments, compounds described herein are superior relative to comparator compounds described in WO2017/120397 because they demonstrate one or more improved properties, such as in vitro activity.
For example, as described herein, certain compounds, including Compound No. 1129485 and Compound No. 1213144, achieved IC50 values of 0.34 and 0.28, respectively, whereas each of comparator compounds Compound No. 1213029, Compound No. 1213030, Compound No. 1213034, and Compound No. 1358035 achieved IC50 values of >10, 8.15, >10, and 8.59, respectively, (see Table 154 in Example 14 for IC50 values). Therefore, certain compounds described herein are more potent than comparator compounds Compound No. 1213029, Compound No. 1213030, Compound No. 1213034, and Compound No. 1358035 in a multiple dose assay.
Compound No. 1194357 is identical to Compound No. 1129485, with the exception that Compound No. 1194357 comprises a conjugate group, and Compound No. 1270705 is identical to Compound No. 1213144, with the exception that Compound No. 1270705 comprises a conjugate group. Therefore, unconjugated versions of Compound No. 1129485 and Compound No. 1270705 are more active than comparator compounds Compound No. 1213029, Compound No. 1213030, Compound No. 1213034, and Compound No. 1358035 in a multiple dose assay.
IX. Certain Hotspot Regions
In certain embodiments, nucleobases in the ranges specified below comprise a hotspot region of FXII nucleic acid. In certain embodiments, modified oligonucleotides that are complementary to a hotspot region of a FXII nucleic acid achieve an average of more than 50% reduction of FXII RNA in vitro in a standard cell assay.
1. Nucleobases 1,899-1,979 of SEO ID NO: 1
In certain embodiments, nucleobases 1,899-1,979 of SEQ ID NO: 1 comprise a hotspot region. In certain embodiments, modified oligonucleotides are complementary to a portion of nucleobases 1,899-1,979 of SEQ ID NO: 1. In certain embodiments, modified oligonucleotides are 20 nucleobases in length. In certain embodiments, modified oligonucleotides are 16 nucleobases in length. In certain embodiments, modified oligonucleotides are gapmers. In certain embodiments, the gapmers are MOE gapmers. In certain embodiments, the gapmers are cEt gapmers. In certain embodiments, the internucleoside linkages of the modified oligonucleotides are phosphorothioate internucleoside linkages and phosphodiester internucleoside linkages. In certain embodiments, each internucleoside linkage is a phosphorothioate internucleoside linkage.
The nucleobase sequences of SEQ ID NOs: 92, 156, 270, 271, 345, 346, 420, 421, 496, 497, 571, 572, 653, 654, 728, 729, 802, 803, 877, 951, 1028, 1103, 1179, 1253, 1330, 1407, 1484, 1560, 1636, 1710, 1711, 1784, 1859, 1860, 1934, 1935, 2008, 2009, 2084, 2085, 2158, 2232, 2233, 2307, 2308, 2393, 2394, 2395, 2396, 2397, 2398, 2461, 2537, 2538, 2614, 2615, 2691, 2692, 2768, 2769, 2845, 2846, 2921, 2922, 2997, 2998, 3073, 3074, 3144, 3145, 3226, 3227, 3303, 3304, 3379, 3380, 3455, 3456, 3531, 3607, 3683, 3759, 3835, 3911, 4948, 4949, 4954, 4955, 4956, 4957, 4961, 4965, 4966, 4967, 4971, 4972, 4976, 4994, 4995, 4996, 4997, 4998, 4999, 5024, 5025, 5026, 5027, 5028, and 5029 are complementary to nucleobases 1,899-1,979 of SEQ ID NO: 1.
The nucleobase sequences of Compound Nos: 1125047, 1125048, 1125049, 1125050, 1125051, 1125074, 1125075, 1125076, 1125077, 1125078, 1129474, 1129475, 1129476, 1129477, 1129478, 1129479, 1129480, 1129481, 1129482, 1129483, 1129484, 1129485, 1129486, 1129487, 1129488, 1129489, 1129490, 1129491, 1129492, 1129493, 1129494, 1129495, 1129496, 1129497, 1129498, 1129499, 1129500, 1129501, 1129502, 1129503, 1129504, 1129505, 1129506, 1131681, 1131682, 1131683, 1131684, 1131685, 1131686, 1131687, 1131688, 1131689, 1131690, 1131691, 1131692, 1131693, 1131694, 1131695, 1131696, 1131697, 1131698, 1131699, 1131700, 1131701, 1131702, 1131703, 1131704, 1131705, 1131706, 1131707, 1131708, 1131709, 1131710, 1131711, 1131712, 1131713, 1131714, 1131715, 1131716, 1131717, 1131718, 1131719, 1131720, 1131721, 1131722, 1131723, 1131724, 1131725, 1206454, 1206455, 1206456, 1206457, 1206458, 1206459, 1206485, 1206486, 1206487, 1206488, 1206489, 1206490, 1206491, 1206492, 1206505, 1206506, 1206507, 1206508, 1206509, 1206957, 1206958, 1206959, 1206960, 1206961, 1206962, 1206963, 1206964, 1206965, 1206966, 1206967, 1206968, 1206969, 1206970, 1206971, 1206972, 1206973, 1206974, 1206975, 1206976, 1206977, 1206978, 1206979, 1206980, 1206981, 1206982, 1206983, 1206985, 1206986, 1206987, 1206988, 1207098, 1207099, 1207100, 1207101, 1207102, 1207103, 1207104, 1207105, 1207106, 1207107, 1207108, 1207109, 1207110, 1207111, 1207112, 1207113, 1207114, 1207115, 1207116, 1207117, 1207118, 1207120, 1207123, 1207125, 1207127, 1207129, 1207131, 1207133, 1207135, 1207137, 1207139, 1207141, 1207228, 1207229, 1207230, 1207231, 1207232, 1207233, 1207234, 1207235, 1207236, 1207237, 1207238, 1207239, 1207240, 1207241, 1207242, 1207243, 1207244, 1207245, 1207246, 1207247, 1207248, 1207249, 1207250, 1207251, 1207252, 1207253, 1207254, 1207256, 1207257, 1207258, 1207259, 1207345, 1207346, 1207347, 1207348, 1207349, 1207350, 1207351, 1207352, 1207353, 1207354, 1207355, 1207356, 1207357, 1207358, 1207359, 1207360, 1207361, 1207362, 1207363, 1207364, 1207365, 1207366, 1207367, 1207368, 1207369, 1207370, 1207371, 1207372, 1207373, 1207374, 1207375, 1207376, 1207462, 1207463, 1207464, 1207465, 1207466, 1207467, 1207468, 1207469, 1207470, 1207471, 1207472, 1207473, 1207474, 1207475, 1207476, 1207477, 1207478, 1207479, 1207480, 1207481, 1207482, 1207483, 1207484, 1207485, 1207486, 1207487, 1207488, 1207489, 1207490, 1207491, 1207492, 1207493, 1213094, 1213095, 1213096, 1213097, 1213098, 1213099, 1213100, 1213101, 1213102, 1213103, 1213104, 1213105, 1213106, 1213107, 1213108, 1213109, 1213110, 1213111, 1213112, 1213113, 1213114, 1213115, 1213116, 1213117, 1213118, 1213119, 1213120, 1213121, 1213122, 1213123, 1213124, 1213125, 1213211, 1213212, 1213213, 1213214, 1213215, 1213216, 1213217, 1213218, 1213219, 1213220, 1213221, 1213222, 1213223, 1213224, 1213225, 1213226, 1213227, 1213228, 1213229, 1213230, 1213231, 1213232, 1213233, 1213234, 1213235, 1213236, 1213237, 1213238, 1213239, 1213240, 1213241, and 1213242 are complementary to nucleobases 1,899-1,979 of SEQ ID NO: 1.
In certain embodiments, modified oligonucleotides complementary to nucleobases 1,899-1,979 of SEQ ID NO: 1 achieve an average of 78% reduction of F12 mRNA in vitro in the standard cell assay.
2. Nucleobases 2,004-2,045 of SEO ID NO: 1
In certain embodiments, nucleobases 2,004-2,045 of SEQ ID NO: 1 comprise a hotspot region. In certain embodiments, modified oligonucleotides are complementary to nucleobases 2,004-2,045 of SEQ ID NO: 1.
The nucleobase sequences of SEQ ID NOs: 95, 159, 162, 163, 164, 165, 347, 422, 498, 573, 953, 1030, 1105, 1255, 1332, 1409, 1486, 1562, 2309, 2463, 2540, 2617, 2694, 2771, 2848, 2924, 3000, 3076, 3147, 3229, 3306, 3382, 3457, 3533, 3609, 3685, 3761, 3837, 3913, 5000, 5001, 5002, 5003, 5004, 5005, 5006, 5031, 5032, 5033, 5034, 5035, 5036, and 5037 are complementary to nucleobases 2,004-2,045 of SEQ ID NO: 1.
The modified oligonucleotides of Compound Nos: 462192, 1125101, 1125102, 1125103, 1125104, 1129526, 1129527, 1129528, 1129529, 1129530, 1129531, 1129532, 1129533, 1129534, 1129535, 1129536, 1129537, 1129538, 1129539, 1129540, 1129541, 1129542, 1129543, 1129544, 1129545, 1131745, 1131746, 1131747, 1131748, 1131749, 1131750, 1131751, 1131752, 1131753, 1131754, 1131755, 1131756, 1131757, 1131758, 1131759, 1131760, 1131761, 1131762, 1131763, 1206993, 1206994, 1206995, 1206996, 1206997, 1206998, 1206999, 1207000, 1207001, 1207002, 1207003, 1207004, 1207005, 1207006, 1207007, 1207008, 1207010, 1207011, 1207012, 1207147, 1207148, 1207149, 1207150, 1207151, 1207152, 1207153, 1207154, 1207155, 1207156, 1207157, 1207158, 1207159, 1207160, 1207161, 1207162, 1207163, 1207164, 1207165, 1207166, 1207264, 1207265, 1207266, 1207267, 1207268, 1207269, 1207270, 1207271, 1207272, 1207273, 1207274, 1207275, 1207276, 1207277, 1207278, 1207279, 1207280, 1207281, 1207282, 1207283, 1207381, 1207382, 1207383, 1207384, 1207385, 1207386, 1207387, 1207388, 1207389, 1207390, 1207391, 1207392, 1207393, 1207394, 1207395, 1207396, 1207397, 1207398, 1207399, 1207400, 1207498, 1207499, 1207500, 1207501, 1207502, 1207503, 1207504, 1207505, 1207506, 1207507, 1207508, 1207509, 1207511, 1207512, 1207513, 1207514, 1207515, 1207516, 1207517, 1213130, 1213131, 1213132, 1213133, 1213134, 1213135, 1213136, 1213137, 1213138, 1213139, 1213140, 1213141, 1213142, 1213143, 1213144, 1213145, 1213146, 1213147, 1213148, 1213149, 1213247, 1213248, 1213249, 1213250, 1213251, 1213252, 1213253, 1213254, 1213255, 1213256, 1213257, 1213258, 1213259, 1213260, 1213261, 1213262, 1213263, 1213264, 1213265, and 1213266 are complementary to nucleobases 2,004-2,045 of SEQ ID NO: 1.
In certain embodiments, modified oligonucleotides complementary to nucleobases 2,004-2,045 of SEQ ID NO: 1 achieve an average of 80% reduction of F12 mRNA in vitro in the standard cell assay.
X. Certain Indications
In certain embodiments, methods comprise administering a pharmaceutical composition described herein to a subject with a thromboembolic condition. Thromboembolic conditions include, but are not limited to, myocardial infarction (MI), stroke, limb ischemia and necrosis, and venous thromboembolism (VTE), including deep vein thrombosis (DVT) and pulmonary embolism (PE).
In certain embodiments, methods comprise administering a pharmaceutical composition described herein to a subject having a risk factor for developing a thromboembolic condition. In certain embodiments, the risk factor is genetic, health-associated, or environmental, or a combination thereof. In certain embodiments, the risk factor is surgery, malignancy, pregnancy, aging, use of oral contraceptives, immobility (including travel-related immobility), sepsis, having a mechanical heart valve, valvular heart disease, atrial fibrillation, atherosclerosis, antiphospholipid syndrome, an inherited clotting disorder (e.g., Factor V Leiden), or an acquired prothrombotic clotting disorder. Identifying a subject with a risk factor for developing a thromboembolic condition may be accomplished by evaluating a subject's medical history and/or by conducting standard clinical tests or assessments.
In certain embodiments, methods comprise administering a pharmaceutical composition described herein to a subject who has been identified as in need of anticoagulation therapy. Non-limiting examples of such subjects include a subject undergoing major orthopedic surgery (e.g., hip/knee replacement or hip fracture surgery) and a subject with atrial fibrillation.
Nonlimiting Disclosure and Incorporation by Reference
Each of the literature and patent publications listed herein is incorporated by reference in its entirety.
While certain compounds, compositions and methods described herein have been described with specificity in accordance with certain embodiments, the following examples serve only to illustrate the compounds described herein and are not intended to limit the same. Each of the references, GenBank accession numbers, and the like recited in the present application is incorporated herein by reference in its entirety.
Although the sequence listing accompanying this filing identifies each sequence as either “RNA” or “DNA” as required, in reality, those sequences may be modified with any combination of chemical modifications. One of skill in the art will readily appreciate that such designation as “RNA” or “DNA” to describe modified oligonucleotides is, in certain instances, arbitrary. For example, an oligonucleotide comprising a nucleoside comprising a 2′-OH sugar moiety and a thymine base could be described as a DNA having a modified sugar (2′-OH in place of one 2′-H of DNA) or as an RNA having a modified base (thymine (methylated uracil) in place of an uracil of RNA). Accordingly, nucleic acid sequences provided herein, including, but not limited to those in the sequence listing, are intended to encompass nucleic acids containing any combination of natural or modified RNA and/or DNA, including, but not limited to such nucleic acids having modified nucleobases. By way of further example and without limitation, an oligomeric compound having the nucleobase sequence “ATCGATCG” encompasses any oligomeric compounds having such nucleobase sequence, whether modified or unmodified, including, but not limited to, such compounds comprising RNA bases, such as those having sequence “AUCGAUCG” and those having some DNA bases and some RNA bases such as “AUCGATCG” and oligomeric compounds having other modified nucleobases, such as “ATmCGAUCG,” wherein mC indicates a cytosine base comprising a methyl group at the 5-position.
Certain compounds described herein (e.g., modified oligonucleotides) have one or more asymmetric center and thus give rise to enantiomers, diastereomers, and other stereoisomeric configurations that may be defined, in terms of absolute stereochemistry, as (R) or (S), as a or R such as for sugar anomers, or as (D) or (L), such as for amino acids, etc. Compounds provided herein that are drawn or described as having certain stereoisomeric configurations include only the indicated compounds. Compounds provided herein that are drawn or described with undefined stereochemistry include all such possible isomers, including their stereorandom and optically pure forms, unless specified otherwise. Likewise, tautomeric forms of the compounds herein are also included unless otherwise indicated. Unless otherwise indicated, compounds described herein are intended to include corresponding salt forms.
The compounds described herein include variations in which one or more atoms are replaced with a non-radioactive isotope or radioactive isotope of the indicated element. For example, compounds herein that comprise hydrogen atoms encompass all possible deuterium substitutions for each of the 1H hydrogen atoms. Isotopic substitutions encompassed by the compounds herein include but are not limited to: 2H or 3H in place of 1H, 13C or 14C in place of 12C, 15N in place of 14N, 17O or 18O in place of 16O, and 33S, 34S, 35S, or 36S in place of 32S. In certain embodiments, non-radioactive isotopic substitutions may impart new properties on the oligomeric compound that are beneficial for use as a therapeutic or research tool. In certain embodiments, radioactive isotopic substitutions may make the compound suitable for research or diagnostic purposes such as imaging.
EXAMPLES
The following examples illustrate certain embodiments of the present disclosure and are not limiting. Moreover, where specific embodiments are provided, the inventors have contemplated generic application of those specific embodiments. For example, disclosure of an oligonucleotide having a particular motif provides reasonable support for additional oligonucleotides having the same or similar motif And, for example, where a particular high-affinity modification appears at a particular position, other high-affinity modifications at the same position are considered suitable, unless otherwise indicated.
Example 1: Effects of 5-10-5 MOE Gapmers on Human FXII RNA In Vitro, Single Dose
Modified oligonucleotides complementary to an FXII nucleic acid were synthesized and tested for their effect on FXII RNA levels in vitro. The modified oligonucleotides were tested in a series of experiments using culture conditions as indicated. The results for each separate experiment are presented in separate tables below.
The modified oligonucleotides in the tables below are 5-10-5 MOE gapmers (i.e., they have a central region of ten 2′-deoxynucleosides flanked on each side by wings, each comprising five 2′-MOE nucleosides). The motif for the gapmers is (from 5′ to 3′): eeeeeddddddddddeeeee; wherein “d” represents a 2′-β-D-deoxyribosyl sugar moiety, and ‘e’ represents a 2′-MOE sugar moiety. The internucleoside linkages throughout each modified oligonucleotide are phosphorothioate linkages. All cytosine nucleobases throughout each modified oligonucleotide are 5-methylcytosines.
“Start site” indicates the 5′-most nucleoside of the target sequence to which the modified oligonucleotide is complementary. “Stop site” indicates the 3′-most nucleoside of the target sequence to which the modified oligonucleotide is complementary. As shown in the tables below, the modified oligonucleotides are 100% complementary to one or more human FXII target sequences, including the human FXII mRNA sequence designated herein as SEQ ID NO: 1 (ENSEMBL ID ENST00000253496.3 from ENSEMBL version 99: January 2020), the human FXII genomic sequence, designated herein as SEQ ID NO: 2 (ENSEMBL ID ENSG00000131187.9 from ENSEML version 99: January 2020), the human FXII genomic sequence, designated herein as SEQ ID NO: 3 (the complement of GENBANK Accession No. NC_000005.10 truncated from truncated from nucleotides 177399001 to 177413000), and the human FXII mRNA sequence designated herein as SEQ ID NO: 4 (GENBANK Accession No. NM_000505.3). ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular target sequence with 100% complementarity.
Cultured Huh7 cells were transfected either with lipofectin, oligofectamine, or by electroporation, as indicated in the tables below. In some cases, Huh7 cells, at a density of 8,000 cells per well, were transfected using lipofectin with 120 nM modified oligonucleotide. In other cases, Huh7 cells, at a density of 5,000 cells per well, were transfected using oligofectamine with 200 nM modified oligonucleotide. In other cases, Huh7 cells, at a density of 20,000 cells per well, were transfected using electroporation with 5000 nM modified oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and FXII RNA levels were measured by quantitative real-time RTPCR. Either human primer probe set RTS2992 (forward sequence GTCAACACTTTCGATTCCACCTT, designated herein as SEQ ID NO: 5; reverse sequence TCCCCGGTGACAGTGAGAAC, designated herein as SEQ ID NO: 6; probe sequence AAGCCCCCAAGGAGCATAAGTACAAAGCTG, designated herein as SEQ ID NO: 7) or human primer probe set RTS40528 (forward sequence GTGCACGGATCCTCCATC, designated herein as SEQ ID NO: 8; reverse sequence CAGCTTGGTCCTCACACAC, designated herein as SEQ ID NO: 9; probe sequence AATCACCCTGGCACGCATCG, designated herein as SEQ ID NO: 10) was used to measure RNA levels. FXII RNA levels were normalized to total RNA content, as measured by RIBOGREEN®. Reduction of FXII RNA is presented in the tables below as percent FXII RNA relative to the amount in untreated control cells (% UTC). The symbol “†” indicates that the modified oligonucleotide is complementary to the target transcript within the amplicon region of the primer probe set. In such instances, additional assays using alternative primer probes must be performed to accurately assess the potency and efficacy of such modified oligonucleotides.
TABLE 1
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with full PS internucleoside
|
linkages (Huh7, lipofectin, 120 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
FXII
SEQ
|
Compound
Start
Stop
Start
Stop
(% UTC)
ID
|
Number
Site
Site
Site
Site
Sequence (5' to 3')
RTS2992
NO
|
|
413577
1
20
1
20
CCAGGAGTCCAGATCAATAG
62
20
|
|
413578
67
86
67
86
TCACCAGCAGGAACCCCAGG
21
21
|
|
413579
92
111
N/A
N/A
GGAATCGAAAGTGTTGACTC
19†
22
|
|
413580
159
178
N/A
N/A
GGTGACAGTGAGAACGACTG
6†
23
|
|
413581
164
183
3515
3534
TCCCCGGTGACAGTGAGAAC
3†
24
|
|
413582
217
236
3568
3587
GGGTACATTTGTGGTACAGC
19
25
|
|
413583
227
246
3578
3597
CGGCCCTTGTGGGTACATTT
30
26
|
|
413584
237
256
3588
3607
AGGGCCTGGCCGGCCCTTGT
67
27
|
|
413585
291
310
3798
3817
GTATCCCCATCGCTGGTCCT
14
28
|
|
413586
325
344
N/A
N/A
TGCAGTGGTCTTTCACTTTC
19
29
|
|
413587
399
418
4207
4226
TTGTGGACAGAGACAGTGGG
27
30
|
|
413588
409
428
4217
4236
CAGTGAGGTGTTGTGGACAG
14
31
|
|
413589
452
471
4397
4416
AGAAGCTGAGGCTCAAAGCA
38
32
|
|
413590
462
481
4407
4426
GAAAAACCGGAGAAGCTGAG
24
33
|
|
413591
515
534
4460
4479
TGGCATCTGGCCACAGCTGC
20
34
|
|
413592
520
539
4465
4484
TGCACTGGCATCTGGCCACA
33
35
|
|
413593
528
547
4473
4492
AGGACCCTTGCACTGGCATC
24
36
|
|
413594
538
557
4483
4502
AGTGGGCATCAGGACCCTTG
22
37
|
|
413595
548
567
4493
4512
AGCCGCTGGCAGTGGGCATC
57
38
|
|
413596
562
581
N/A
N/A
AGGCCTGGCTGGCCAGCCGC
112
39
|
|
413597
567
586
N/A
N/A
GCGGCAGGCCTGGCTGGCCA
31
40
|
|
413598
572
591
N/A
N/A
TTGGTGCGGCAGGCCTGGCT
48
41
|
|
413599
587
606
4671
4690
CCATGGAGGCACGGGTTGGT
19
42
|
|
413600
608
627
4692
4711
TCCACCTCTAGGCAGCGACC
19
43
|
|
413601
626
645
4710
4729
TGGCACAGGCGGTGGCCCTC
39
44
|
|
413602
695
714
4924
4943
CCGCGGCCATCATAGCAGCT
24
45
|
|
413603
700
719
4929
4948
TGAGCCCGCGGCCATCATAG
33
46
|
|
413604
705
724
4934
4953
GTAGCTGAGCCCGCGGCCAT
28
47
|
|
413605
713
732
4942
4961
AGGCCGCGGTAGCTGAGCCC
38
48
|
|
413606
718
737
4947
4966
TGGCCAGGCCGCGGTAGCTG
30
49
|
|
413607
723
742
4952
4971
GGTCCTGGCCAGGCCGCGGT
23
50
|
|
413608
746
765
4975
4994
TGACAGGGCGCACCCGAGAG
33
51
|
|
413609
751
770
4980
4999
ACGGCTGACAGGGCGCACCC
28
52
|
|
413610
808
827
5037
5056
GTCCCCAGTTCCGCGCTTGC
21
53
|
|
413611
818
837
5047
5066
TGGCCGCCCAGTCCCCAGTT
29
54
|
|
413612
839
858
N/A
N/A
TCCGGGTTCCGGCAGAAGGC
20
55
|
|
413613
863
882
5177
5196
AAGCACCACGGGCGGATGTC
29
56
|
|
413614
871
890
5185
5204
TCAGCACGAAGCACCACGGG
13
57
|
|
413615
978
997
5292
5311
CATGAGTGGGACATGAAGCC
38
58
|
|
413616
1119
1138
5538
5557
GAGCCGCTGCCCGCAGCTCA
31
59
|
|
413617
1132
1151
5551
5570
ACAGACTCTTGCGGAGCCGC
19
60
|
|
413618
1137
1156
5556
5575
CGAAGACAGACTCTTGCGGA
22
61
|
|
413619
1142
1161
5561
5580
GTCATCGAAGACAGACTCTT
28
62
|
|
413620
1211
1230
5630
5649
TGGCCCCAGTACAGCGCGGC
27
63
|
|
413621
1231
1250
5650
5669
GGCTGCCGGCGCAGAAACTG
32
64
|
|
413622
1236
1255
5655
5674
GATGAGGCTGCCGGCGCAGA
25
65
|
|
413623
1283
1302
N/A
N/A
GGCCGGTCCTGCAGGCAGTG
28
66
|
|
413624
1304
1323
5964
5983
ACCGTCAGATCCTCGGGTGC
33
67
|
|
413625
1379
1398
6039
6058
TGCAAGCGGTAGGAGCGCAC
32
68
|
|
413626
1429
1448
N/A
N/A
GGCGCAACAGAGCCAGGTCG
100
69
|
|
413627
1455
1474
6198
6217
GCAGCTGCCGTCCGCATCCT
28
70
|
|
413628
1466
1485
6209
6228
GACAGGAGCGCGCAGCTGCC
25
71
|
|
413629
1492
1511
6235
6254
GCAGGCACACCGGCTGAACG
19
72
|
|
413630
1502
1521
6245
6264
GCGCCGCTTGGCAGGCACAC
28
73
|
|
413631
1559
1578
6302
6321
TCGAACTGGTGGCCCCAGCC
30
74
|
|
413632
1679
1698
6977
6996
CCTGCGCAGAGCATGCCGGG
50
75
|
|
413633
1684
1703
6982
7001
GGAACCCTGCGCAGAGCATG
17
76
|
|
413634
1689
1708
6987
7006
CTCGAGGAACCCTGCGCAGA
36
77
|
|
413635
1695
1714
6993
7012
GCCGCCCTCGAGGAACCCTG
14
78
|
|
413636
1743
1762
7131
7150
GTCCTCACACACCAGCGGGC
16
79
|
|
413637
1748
1767
7136
7155
GCTTGGTCCTCACACACCAG
18
80
|
|
413638
1774
1793
7162
7181
CTTGCAGGGTGAGCCGGCGC
45
81
|
|
413639
1779
1798
7167
7186
GATGCCTTGCAGGGTGAGCC
29
82
|
|
413640
1809
1828
7197
7216
GCGGTCACCACAGCCCGATC
25
83
|
|
413641
1814
1833
7202
7221
TTGTTGCGGTCACCACAGCC
23
84
|
|
413642
1820
1839
7208
7227
CCTGGCTTGTTGCGGTCACC
141
85
|
|
413643
1839
1858
7227
7246
GGCCACATCGGTGTAGACGC
16
86
|
|
413644
1844
1863
7232
7251
TAGTAGGCCACATCGGTGTA
32
87
|
|
413645
1849
1868
7237
7256
CCAGGTAGTAGGCCACATCG
21
88
|
|
413646
1854
1873
7242
7261
CCAGGCCAGGTAGTAGGCCA
38
89
|
|
413647
1859
1878
7247
7266
CGGATCCAGGCCAGGTAGTA
33
90
|
|
413648
1891
1910
7279
7298
AGTCCCTGAGCAATCAGGAA
19
91
|
|
413649
1901
1920
7289
7308
GGGAAAGATGAGTCCCTGAG
14
92
|
|
413650
1992
2011
7380
7399
ATCCTGGCGCGGAGCTGGCC
13
93
|
|
413651
2002
2021
7390
7409
TTCCTGCGCCATCCTGGCGC
23
94
|
|
413652
2012
2031
7400
7419
CTTTATTGAGTTCCTGCGCC
15
95
|
|
413653
2022
2041
7410
7429
TTTCAAAGCACTTTATTGAG
11
96
|
|
413654
2027
2046
7415
7434
AGCATTTTCAAAGCACTTTA
23
97
|
|
TABLE 2
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers
|
with full PS internucleoside linkages (Huh7,
|
oligofectamine, 200 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
FXII
SEQ
|
Compound
Start
Stop
Start
Stop
(% UTC)
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
RTS2992
NO
|
|
413627
1455
1474
6198
6217
GCAGCTGCCGTCCGCATCCT
27
70
|
|
413633
1684
1703
6982
7001
GGAACCCTGCGCAGAGCATG
41
76
|
|
413635
1695
1714
6993
7012
GCCGCCCTCGAGGAACCCTG
34
78
|
|
413643
1839
1858
7227
7246
GGCCACATCGGTGTAGACGC
23
86
|
|
413648
1891
1910
7279
7298
AGTCCCTGAGCAATCAGGAA
33
91
|
|
413649
1901
1920
7289
7308
GGGAAAGATGAGTCCCTGAG
18
92
|
|
413652
2012
2031
7400
7419
CTTTATTGAGTTCCTGCGCC
14
95
|
|
462131
1128
1147
5547
5566
ACTCTTGCGGAGCCGCTGCC
23
98
|
|
462132
1145
1164
5564
5583
CGGGTCATCGAAGACAGACT
39
99
|
|
462133
1148
1167
5567
5586
ACGCGGGTCATCGAAGACAG
24
100
|
|
462134
1151
1170
5570
5589
ACGACGCGGGTCATCGAAGA
52
101
|
|
462135
1154
1173
5573
5592
CCAACGACGCGGGTCATCGA
57
102
|
|
462136
1157
1176
5576
5595
CCGCCAACGACGCGGGTCAT
51
103
|
|
462137
1160
1179
5579
5598
AGCCCGCCAACGACGCGGGT
59
104
|
|
462138
1163
1182
5582
5601
ACCAGCCCGCCAACGACGCG
45
105
|
|
462139
1166
1185
5585
5604
GCCACCAGCCCGCCAACGAC
46
106
|
|
462140
1202
1221
5621
5640
TACAGCGCGGCGATGTAGGG
44
107
|
|
462141
1205
1224
5624
5643
CAGTACAGCGCGGCGATGTA
61
108
|
|
462142
1208
1227
5627
5646
CCCCAGTACAGCGCGGCGAT
30
109
|
|
462143
1214
1233
5633
5652
CTGTGGCCCCAGTACAGCGC
28
110
|
|
462144
1217
1236
5636
5655
AAACTGTGGCCCCAGTACAG
42
111
|
|
462145
1223
1242
5642
5661
GCGCAGAAACTGTGGCCCCA
20
112
|
|
462146
1226
1245
5645
5664
CCGGCGCAGAAACTGTGGCC
41
113
|
|
462147
1239
1258
5658
5677
GGCGATGAGGCTGCCGGCGC
22
114
|
|
462148
1374
1393
6034
6053
GCGGTAGGAGCGCACGGCCA
32
115
|
|
462149
1432
1451
N/A
N/A
GAAGGCGCAACAGAGCCAGG
61
116
|
|
462150
1435
1454
N/A
N/A
CCTGAAGGCGCAACAGAGCC
56
117
|
|
462151
1438
1457
6181
6200
CCTCCTGAAGGCGCAACAGA
48
118
|
|
462152
1444
1463
6187
6206
CCGCATCCTCCTGAAGGCGC
25
119
|
|
462153
1447
1466
6190
6209
CGTCCGCATCCTCCTGAAGG
50
120
|
|
462154
1450
1469
6193
6212
TGCCGTCCGCATCCTCCTGA
37
121
|
|
462155
1453
1472
6196
6215
AGCTGCCGTCCGCATCCTCC
39
122
|
|
462156
1454
1473
6197
6216
CAGCTGCCGTCCGCATCCTC
36
123
|
|
462157
1456
1475
6199
6218
CGCAGCTGCCGTCCGCATCC
22
124
|
|
462158
1457
1476
6200
6219
GCGCAGCTGCCGTCCGCATC
31
125
|
|
462159
1458
1477
6201
6220
CGCGCAGCTGCCGTCCGCAT
28
126
|
|
462160
1461
1480
6204
6223
GAGCGCGCAGCTGCCGTCCG
41
127
|
|
462161
1469
1488
6212
6231
GGCGACAGGAGCGCGCAGCT
25
128
|
|
462162
1472
1491
6215
6234
TAAGGCGACAGGAGCGCGCA
41
129
|
|
462163
1495
1514
6238
6257
TTGGCAGGCACACCGGCTGA
32
130
|
|
462164
1498
1517
6241
6260
CGCTTGGCAGGCACACCGGC
36
131
|
|
462165
1518
1537
6261
6280
CTCGGAGGGTCGCGCGGCGC
28
132
|
|
462166
1562
1581
6305
6324
CCCTCGAACTGGTGGCCCCA
42
133
|
|
462167
1596
1615
6894
6913
CGCCTCCTGCAGGAAGCTGG
33
134
|
|
462168
1599
1618
6897
6916
CTGCGCCTCCTGCAGGAAGC
38
135
|
|
462169
1602
1621
6900
6919
TACCTGCGCCTCCTGCAGGA
67
136
|
|
462170
1682
1701
6980
6999
AACCCTGCGCAGAGCATGCC
28
137
|
|
462171
1685
1704
6983
7002
AGGAACCCTGCGCAGAGCAT
36
138
|
|
462172
1686
1705
6984
7003
GAGGAACCCTGCGCAGAGCA
27
139
|
|
462173
1692
1711
6990
7009
GCCCTCGAGGAACCCTGCGC
38
140
|
|
462174
1693
1712
6991
7010
CGCCCTCGAGGAACCCTGCG
39
141
|
|
462175
1697
1716
6995
7014
GTGCCGCCCTCGAGGAACCC
44
142
|
|
462176
1698
1717
6996
7015
GGTGCCGCCCTCGAGGAACC
19
143
|
|
462177
1702
1721
7000
7019
CATCGGTGCCGCCCTCGAGG
50
144
|
|
462178
1840
1859
7228
7247
AGGCCACATCGGTGTAGACG
30
145
|
|
462179
1841
1860
7229
7248
TAGGCCACATCGGTGTAGAC
36
146
|
|
462180
1889
1908
7277
7296
TCCCTGAGCAATCAGGAAAC
31
147
|
|
462181
1890
1909
7278
7297
GTCCCTGAGCAATCAGGAAA
29
148
|
|
462182
1892
1911
7280
7299
GAGTCCCTGAGCAATCAGGA
39
149
|
|
462183
1893
1912
7281
7300
TGAGTCCCTGAGCAATCAGG
16
150
|
|
462184
1899
1918
7287
7306
GAAAGATGAGTCCCTGAGCA
34
151
|
|
462185
1900
1919
7288
7307
GGAAAGATGAGTCCCTGAGC
24
152
|
|
462186
1902
1921
7290
7309
AGGGAAAGATGAGTCCCTGA
25
153
|
|
462187
1903
1922
7291
7310
GAGGGAAAGATGAGTCCCTG
24
154
|
|
462188
1904
1923
7292
7311
GGAGGGAAAGATGAGTCCCT
21
155
|
|
462189
1947
1966
7335
7354
TGCCTTCCATGCCCCAGCCA
17
156
|
|
462190
1995
2014
7383
7402
GCCATCCTGGCGCGGAGCTG
23
157
|
|
462191
1998
2017
7386
7405
TGCGCCATCCTGGCGCGGAG
50
158
|
|
462192
2005
2024
7393
7412
GAGTTCCTGCGCCATCCTGG
13
159
|
|
462193
2008
2027
7396
7415
ATTGAGTTCCTGCGCCATCC
17
160
|
|
462194
2011
2030
7399
7418
TTTATTGAGTTCCTGCGCCA
22
161
|
|
462195
2013
2032
7401
7420
ACTTTATTGAGTTCCTGCGC
16
162
|
|
462196
2014
2033
7402
7421
CACTTTATTGAGTTCCTGCG
8
163
|
|
462197
2015
2034
7403
7422
GCACTTTATTGAGTTCCTGC
6
164
|
|
462198
2018
2037
7406
7425
AAAGCACTTTATTGAGTTCC
7
165
|
|
462199
2030
2049
7418
7437
CTCAGCATTTTCAAAGCACT
22
166
|
|
TABLE 3
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with full PS internucleoside linkages (Huh7,
|
oligofectamine, 200 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
FXII
SEQ
|
Compound
Start
Stop
Start
Stop
(% UTC)
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
RTS2992
NO
|
|
413582
217
236
3568
3587
GGGTACATTTGTGGTACAGC
42
5
|
|
413592
520
539
4465
4484
TGCACTGGCATCTGGCCACA
57
35
|
|
413610
808
827
5037
5056
GTCCCCAGTTCCGCGCTTGC
45
43
|
|
413611
818
837
5047
5066
TGGCCGCCCAGTCCCCAGTT
77
54
|
|
413612
839
858
N/A
N/A
TCCGGGTTCCGGCAGAAGGC
68
55
|
|
413652
2012
2031
7400
7419
CTTTATTGAGTTCCTGCGCC
30
95
|
|
462056
55
74
55
74
ACCCCAGGAGCAGCAGAGCC
58
167
|
|
462057
58
77
58
77
GGAACCCCAGGAGCAGCAGA
67
168
|
|
462058
61
80
61
80
GCAGGAACCCCAGGAGCAGC
62
169
|
|
462059
64
83
64
83
CCAGCAGGAACCCCAGGAGC
76
170
|
|
462060
95
114
N/A
N/A
GGTGGAATCGAAAGTGTTGA
65†
171
|
|
462061
98
117
N/A
N/A
CAAGGTGGAATCGAAAGTGT
63†
172
|
|
462062
137
156
504
523
TGCTCTTCAGCTTTGTACTT
23†
173
|
|
462063
140
159
507
526
GTGTGCTCTTCAGCTTTGTA
6†
174
|
|
462064
143
162
510
529
ACTGTGTGCTCTTCAGCTTT
10†
175
|
|
462065
146
165
N/A
N/A
ACGACTGTGTGCTCTTCAGC
9†
176
|
|
462067
155
174
N/A
N/A
ACAGTGAGAACGACTGTGTG
30†
177
|
|
462068
167
186
3518
3537
GGCTCCCCGGTGACAGTGAG
13†
178
|
|
462069
170
189
3521
3540
CAGGGCTCCCCGGTGACAGT
14†
179
|
|
462070
173
192
3524
3543
TGGCAGGGCTCCCCGGTGAC
16†
180
|
|
462071
176
195
3527
3546
AAGTGGCAGGGCTCCCCGGT
70†
181
|
|
462072
215
234
3566
3585
GTACATTTGTGGTACAGCTG
16
182
|
|
462073
216
235
3567
3586
GGTACATTTGTGGTACAGCT
24
183
|
|
462074
219
238
3570
3589
GTGGGTACATTTGTGGTACA
38
184
|
|
462075
223
242
3574
3593
CCTTGTGGGTACATTTGTGG
48
185
|
|
462076
230
249
3581
3600
GGCCGGCCCTTGTGGGTACA
65
186
|
|
462077
315
334
3822
3841
TTTCACTTTCTTGGGCTCCA
46
187
|
|
462078
318
337
N/A
N/A
GTCTTTCACTTTCTTGGGCT
59
188
|
|
462079
321
340
N/A
N/A
GTGGTCTTTCACTTTCTTGG
69
189
|
|
462080
448
467
4393
4412
GCTGAGGCTCAAAGCACTTC
42
190
|
|
462081
455
474
4400
4419
CGGAGAAGCTGAGGCTCAAA
71
191
|
|
462082
458
477
4403
4422
AACCGGAGAAGCTGAGGCTC
60
192
|
|
462083
465
484
4410
4429
GTGGAAAAACCGGAGAAGCT
46
193
|
|
462084
486
505
4431
4450
TCTATACCATATCTCATTCT
57
194
|
|
462085
507
526
4452
4471
GGCCACAGCTGCTTGCTCAG
44
195
|
|
462086
518
537
4463
4482
CACTGGCATCTGGCCACAGC
51
196
|
|
462087
519
538
4464
4483
GCACTGGCATCTGGCCACAG
62
197
|
|
462088
521
540
4466
4485
TTGCACTGGCATCTGGCCAC
37
198
|
|
462089
522
541
4467
4486
CTTGCACTGGCATCTGGCCA
55
199
|
|
462090
523
542
4468
4487
CCTTGCACTGGCATCTGGCC
81
200
|
|
462091
534
553
4479
4498
GGCATCAGGACCCTTGCACT
35
201
|
|
462092
541
560
4486
4505
GGCAGTGGGCATCAGGACCC
59
202
|
|
462093
544
563
4489
4508
GCTGGCAGTGGGCATCAGGA
85
203
|
|
462094
575
594
N/A
N/A
GGGTTGGTGCGGCAGGCCTG
68
204
|
|
462095
578
597
4662
4681
CACGGGTTGGTGCGGCAGGC
65
205
|
|
462096
581
600
4665
4684
AGGCACGGGTTGGTGCGGCA
65
206
|
|
462097
584
603
4668
4687
TGGAGGCACGGGTTGGTGCG
105
207
|
|
462098
590
609
4674
4693
CCCCCATGGAGGCACGGGTT
32
208
|
|
462099
593
612
4677
4696
CGACCCCCATGGAGGCACGG
51
209
|
|
462100
596
615
4680
4699
CAGCGACCCCCATGGAGGCA
59
210
|
|
462101
599
618
4683
4702
AGGCAGCGACCCCCATGGAG
51
211
|
|
462102
602
621
4686
4705
TCTAGGCAGCGACCCCCATG
50
212
|
|
462103
605
624
4689
4708
ACCTCTAGGCAGCGACCCCC
59
213
|
|
462104
611
630
4695
4714
CCCTCCACCTCTAGGCAGCG
60
214
|
|
462105
614
633
4698
4717
TGGCCCTCCACCTCTAGGCA
47
215
|
|
462106
620
639
4704
4723
AGGCGGTGGCCCTCCACCTC
73
216
|
|
462107
623
642
4707
4726
CACAGGCGGTGGCCCTCCAC
27
217
|
|
462108
629
648
4713
4732
CAGTGGCACAGGCGGTGGCC
45
218
|
|
462109
632
651
4716
4735
GGGCAGTGGCACAGGCGGTG
54
219
|
|
462110
708
727
4937
4956
GCGGTAGCTGAGCCCGCGGC
81
220
|
|
462111
726
745
4955
4974
CGTGGTCCTGGCCAGGCCGC
61
221
|
|
462112
803
822
5032
5051
CAGTTCCGCGCTTGCTCGGC
51
222
|
|
462113
806
825
5035
5054
CCCCAGTTCCGCGCTTGCTC
44
223
|
|
462114
807
826
5036
5055
TCCCCAGTTCCGCGCTTGCT
69
224
|
|
462115
809
828
5038
5057
AGTCCCCAGTTCCGCGCTTG
55
225
|
|
462116
810
829
5039
5058
CAGTCCCCAGTTCCGCGCTT
56
226
|
|
462117
811
830
5040
5059
CCAGTCCCCAGTTCCGCGCT
40
227
|
|
462118
814
833
5043
5062
CGCCCAGTCCCCAGTTCCGC
60
228
|
|
462119
817
836
5046
5065
GGCCGCCCAGTCCCCAGTTC
57
229
|
|
462120
820
839
5049
5068
CGTGGCCGCCCAGTCCCCAG
43
230
|
|
462121
838
857
N/A
N/A
CCGGGTTCCGGCAGAAGGCG
59
231
|
|
462122
841
860
N/A
N/A
TGTCCGGGTTCCGGCAGAAG
95
232
|
|
462123
842
861
N/A
N/A
TTGTCCGGGTTCCGGCAGAA
96
233
|
|
462124
866
885
5180
5199
ACGAAGCACCACGGGCGGAT
85
234
|
|
462125
1002
1021
5316
5335
CTTCGGCGGTGCCGGCTGCG
57
235
|
|
462126
1122
1141
5541
5560
GCGGAGCCGCTGCCCGCAGC
80
236
|
|
462127
1125
1144
5544
5563
CTTGCGGAGCCGCTGCCCGC
79
237
|
|
TABLE 4
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with full PS internucleoside linkages (Huh7,
|
electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
FXII
SEQ
|
Compound
Start
Stop
Start
Stop
(% UTC)
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
RTS40528
NO
|
|
413612
839
858
N/A
N/A
TCCGGGTTCCGGCAGAAGGC
31
55
|
|
462073
216
235
3567
3586
GGTACATTTGTGGTACAGCT
10
183
|
|
462192
2005
2024
7393
7412
GAGTTCCTGCGCCATCCTGG
22
159
|
|
1124163
32
51
32
51
ATGGCATCCGTCCGTTGGTC
70
238
|
|
1124191
78
97
78
97
TGACTCCAAGCTCACCAGCA
79
239
|
|
1124219
135
154
502
521
CTCTTCAGCTTTGTACTTAT
60
240
|
|
1124245
171
190
3522
3541
GCAGGGCTCCCCGGTGACAG
42
241
|
|
1124268
225
244
3576
3595
GCCCTTGTGGGTACATTTGT
79
242
|
|
1124296
290
309
3797
3816
TATCCCCATCGCTGGTCCTG
31
243
|
|
1124324
331
350
N/A
N/A
GTTTGCTGCAGTGGTCTTTC
25
244
|
|
1124352
404
423
4212
4231
AGGTGTTGTGGACAGAGACA
60
245
|
|
1124380
446
465
4391
4410
TGAGGCTCAAAGCACTTCTC
43
246
|
|
1124407
489
508
4434
4453
AGTTCTATACCATATCTCAT
32
247
|
|
1124435
532
551
4477
4496
CATCAGGACCCTTGCACTGG
54
248
|
|
1124462
566
585
N/A
N/A
CGGCAGGCCTGGCTGGCCAG
33
249
|
|
1124489
606
625
4690
4709
CACCTCTAGGCAGCGACCCC
21
250
|
|
1124517
653
672
4737
4756
AAGGCTCCGGTGTAGCCCAC
48
251
|
|
1124545
709
728
4938
4957
CGCGGTAGCTGAGCCCGCGG
65
252
|
|
1124572
762
781
4991
5010
CTCCGAGGCCCACGGCTGAC
39
253
|
|
1124620
902
921
5216
5235
TCGCAGTACTCCCAGCTCAG
19
254
|
|
1124648
993
1012
5307
5326
TGCCGGCTGCGCGGGCATGA
37
255
|
|
1124676
1071
1090
5490
5509
CTGCTCCCGCTTCGCCGGCA
79
256
|
|
1124704
1116
1135
5535
5554
CCGCTGCCCGCAGCTCAGTG
50
257
|
|
1124730
1152
1171
5571
5590
AACGACGCGGGTCATCGAAG
27
258
|
|
1124757
1224
1243
5643
5662
GGCGCAGAAACTGTGGCCCC
32
259
|
|
1124784
1311
1330
5971
5990
GAGCACCACCGTCAGATCCT
30
260
|
|
1124812
1380
1399
6040
6059
GTGCAAGCGGTAGGAGCGCA
48
261
|
|
1124839
1459
1478
6202
6221
GCGCGCAGCTGCCGTCCGCA
27
262
|
|
1124863
1548
1567
6291
6310
GCCCCAGCCGGCCACCTGGC
149
263
|
|
1124890
1578
1597
N/A
N/A
GGCATATTCCTCCGCCCCCT
37
264
|
|
1124917
1620
1639
6918
6937
CTCCAGGGAGAGGAACGGTA
116
265
|
|
1124943
1746
1765
7134
7153
TTGGTCCTCACACACCAGCG
10†
266
|
|
1124970
1793
1812
7181
7200
GATCCCCAGCTGATGATGCC
94
267
|
|
1124996
1823
1842
7211
7230
ACGCCTGGCTTGTTGCGGTC
24
268
|
|
1125021
1870
1889
7258
7277
CGGTGTGCTCCCGGATCCAG
50
269
|
|
1125047
1915
1934
7303
7322
GGAATCACCAAGGAGGGAAA
14
270
|
|
1125074
1950
1969
7338
7357
TCTTGCCTTCCATGCCCCAG
18
271
|
|
1125122
N/A
N/A
94
113
CACTCACCGAAAGTGTTGAC
40
272
|
|
1125150
N/A
N/A
178
197
CCACAGGTCATGAGCAGAGG
63
273
|
|
1125178
N/A
N/A
274
293
TGCCCTTGTATCCACCCAGT
71
274
|
|
1125206
N/A
N/A
368
387
CCCAGAAATGCAGAGATTTC
103
275
|
|
1125234
N/A
N/A
457
476
AATCTACAAGGGAGAGAAGA
51
276
|
|
1125262
N/A
N/A
603
622
GGCACTAGACTAGACTGCCC
33
277
|
|
1125290
N/A
N/A
690
709
CTTAGACACAGCCCATACCC
25
278
|
|
1125318
N/A
N/A
859
878
ATTGAAGGCCCTGGCCTCTG
37
279
|
|
1125346
N/A
N/A
957
976
TCAAATCCCTCGCCCAGAGT
56
280
|
|
1125374
N/A
N/A
1064
1083
CAGGTGATGGGCTGAATACC
123
281
|
|
1125402
N/A
N/A
1197
1216
AATTTAGTGTCTGGAAGAAT
44
282
|
|
1125430
N/A
N/A
1575
1594
CCCCACCAAGTCAGGCCCAC
66
283
|
|
1125458
N/A
N/A
1747
1766
TGGACGGACAGAGAGGAGGG
127
284
|
|
1125486
N/A
N/A
2123
2142
AGGACCAAACCAGAACTAGG
119
285
|
|
1125514
N/A
N/A
2253
2272
TGTCTTCTTCAGAGGAATTG
47
286
|
|
1125542
N/A
N/A
2341
2360
GGCTCACAGGAACCTAATAC
36
287
|
|
1125570
N/A
N/A
2479
2498
GATTCAGGTGTGTGCTATAG
13
288
|
|
1125598
N/A
N/A
2609
2628
GTTTTTCACACTTTGTGCTT
31
289
|
|
1125626
N/A
N/A
2734
2753
GGGCATGCACAGAGCAGTGA
47
290
|
|
1125654
N/A
N/A
2832
2851
CTCATTCTTCTCTGGTTCTA
32
291
|
|
1125682
N/A
N/A
2939
2958
TAGGCACTTAATAATACTTG
51
292
|
|
1125710
N/A
N/A
3042
3061
GGAGTCAGATAGCTGGAGTA
163
293
|
|
1125738
N/A
N/A
3436
3455
CTCAGGGTCTGGTCAGGAAA
62
294
|
|
1125766
N/A
N/A
3631
3650
CAGGCCCCTGCTCCAACTCC
107
295
|
|
1125794
N/A
N/A
3743
3762
GAGACAAGGCTTCCCTGCTC
38
296
|
|
1125822
N/A
N/A
3889
3908
GATACCAGGAGAGTAATGAG
103
297
|
|
1125850
N/A
N/A
4015
4034
AAGCTTGGTTTACCCACCTG
94
298
|
|
1125878
N/A
N/A
4122
4141
GAGAGATGGACATGGTGGAA
79
299
|
|
1125906
N/A
N/A
4310
4329
CCAGCAACCTATTCTGTAGG
47
300
|
|
1125934
N/A
N/A
4535
4554
CTCCCTGGCCCGTTCCCAAC
30
301
|
|
1125962
N/A
N/A
4632
4651
AACGCAGTGAGCCACCCCTG
61
302
|
|
1125990
N/A
N/A
4859
4878
CTGGGCTCTCCTGCCTCCCT
80
303
|
|
1126018
N/A
N/A
5383
5402
TTCCCCCCCCCACTTCCTAA
187
304
|
|
1126046
N/A
N/A
5751
5770
GAGACGGAGGAGCCGCGGCC
120
305
|
|
1126074
N/A
N/A
6158
6177
AACCCGGGCGGAGAGGAGCG
170
306
|
|
1126102
N/A
N/A
6373
6392
GGCTTCTTCCGCCTAACCCA
111
307
|
|
1126130
N/A
N/A
6481
6500
TGAATCCCAGGCCCTGGGAT
73
308
|
|
1126158
N/A
N/A
6621
6640
GAGTCGCAGAACCTGGCTCC
84
309
|
|
1126186
N/A
N/A
6712
6731
ATTTCATAGGCAAGGAGGCT
53
310
|
|
1126214
N/A
N/A
6839
6858
CTTCACACCCCATCTGACAA
82
311
|
|
1126242
N/A
N/A
7106
7125
GAATCACCCTGGGTCGGAAA
43†
312
|
|
TABLE 5
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with full PS internucleoside linkages (Huh7,
|
electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
FXII
SEQ
|
Compound
Start
Stop
Start
Stop
(% UTC)
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
RTS40528
NO
|
|
413643
1839
1858
7227
7246
GGCCACATCGGTGTAGACGC
14
86
|
|
462073
216
235
3567
3586
GGTACATTTGTGGTACAGCT
36
183
|
|
626671
840
859
N/A
N/A
GTCCGGGTTCCGGCAGAAGG
19
313
|
|
1124164
33
52
33
52
CATGGCATCCGTCCGTTGGT
24
314
|
|
1124192
79
98
79
98
TTGACTCCAAGCTCACCAGC
95
315
|
|
1124220
136
155
503
522
GCTCTTCAGCTTTGTACTTA
93
316
|
|
1124246
172
191
3523
3542
GGCAGGGCTCCCCGGTGACA
36
317
|
|
1124269
226
245
3577
3596
GGCCCTTGTGGGTACATTTG
34
318
|
|
1124297
292
311
3799
3818
AGTATCCCCATCGCTGGTCC
24
319
|
|
1124325
332
351
N/A
N/A
TGTTTGCTGCAGTGGTCTTT
92
320
|
|
1124353
405
424
4213
4232
GAGGTGTTGTGGACAGAGAC
24
321
|
|
1124381
447
466
4392
4411
CTGAGGCTCAAAGCACTTCT
27
322
|
|
1124408
493
512
4438
4457
GCTCAGTTCTATACCATATC
18
323
|
|
1124436
533
552
4478
4497
GCATCAGGACCCTTGCACTG
78
324
|
|
1124463
568
587
N/A
N/A
TGCGGCAGGCCTGGCTGGCC
65
325
|
|
1124490
607
626
4691
4710
CCACCTCTAGGCAGCGACCC
17
326
|
|
1124518
654
673
4738
4757
GAAGGCTCCGGTGTAGCCCA
75
327
|
|
1124546
710
729
4939
4958
CCGCGGTAGCTGAGCCCGCG
170
328
|
|
1124573
763
782
4992
5011
CCTCCGAGGCCCACGGCTGA
27
329
|
|
1124621
903
922
5217
5236
GTCGCAGTACTCCCAGCTCA
23
330
|
|
1124649
994
1013
5308
5327
GTGCCGGCTGCGCGGGCATG
42
331
|
|
1124677
1072
1091
5491
5510
GCTGCTCCCGCTTCGCCGGC
23
332
|
|
1124705
1117
1136
5536
5555
GCCGCTGCCCGCAGCTCAGT
40
333
|
|
1124731
1153
1172
5572
5591
CAACGACGCGGGTCATCGAA
64
334
|
|
1124758
1225
1244
5644
5663
CGGCGCAGAAACTGTGGCCC
35
335
|
|
1124785
1312
1331
5972
5991
CGAGCACCACCGTCAGATCC
30
336
|
|
1124813
1381
1400
6041
6060
CGTGCAAGCGGTAGGAGCGC
18
337
|
|
1124840
1460
1479
6203
6222
AGCGCGCAGCTGCCGTCCGC
59
338
|
|
1124864
1549
1568
6292
6311
GGCCCCAGCCGGCCACCTGG
54
339
|
|
1124891
1579
1598
N/A
N/A
TGGCATATTCCTCCGCCCCC
34
340
|
|
1124918
1621
1640
6919
6938
GCTCCAGGGAGAGGAACGGT
84
341
|
|
1124944
1747
1766
7135
7154
CTTGGTCCTCACACACCAGC
7†
342
|
|
1124971
1794
1813
7182
7201
CGATCCCCAGCTGATGATGC
66
343
|
|
1125022
1871
1890
7259
7278
ACGGTGTGCTCCCGGATCCA
26
344
|
|
1125048
1916
1935
7304
7323
CGGAATCACCAAGGAGGGAA
35
345
|
|
1125075
1951
1970
7339
7358
ATCTTGCCTTCCATGCCCCA
23
346
|
|
1125101
2006
2025
7394
7413
TGAGTTCCTGCGCCATCCTG
7
347
|
|
1125123
N/A
N/A
97
116
CAGCACTCACCGAAAGTGTT
67
348
|
|
1125151
N/A
N/A
181
200
GACCCACAGGTCATGAGCAG
50
349
|
|
1125179
N/A
N/A
277
296
CTGTGCCCTTGTATCCACCC
67
350
|
|
1125207
N/A
N/A
371
390
CAGCCCAGAAATGCAGAGAT
45
351
|
|
1125235
N/A
N/A
460
479
TGGAATCTACAAGGGAGAGA
125
352
|
|
1125263
N/A
N/A
606
625
GTAGGCACTAGACTAGACTG
129
353
|
|
1125291
N/A
N/A
693
712
GCACTTAGACACAGCCCATA
69
354
|
|
1125319
N/A
N/A
862
881
CCCATTGAAGGCCCTGGCCT
38
355
|
|
1125347
N/A
N/A
960
979
AACTCAAATCCCTCGCCCAG
77
356
|
|
1125375
N/A
N/A
1067
1086
CACCAGGTGATGGGCTGAAT
86
357
|
|
1125403
N/A
N/A
1200
1219
GGTAATTTAGTGTCTGGAAG
130
358
|
|
1125431
N/A
N/A
1578
1597
CAACCCCACCAAGTCAGGCC
74
359
|
|
1125459
N/A
N/A
1750
1769
GGATGGACGGACAGAGAGGA
60
360
|
|
1125487
N/A
N/A
2126
2145
CCAAGGACCAAACCAGAACT
84
361
|
|
1125515
N/A
N/A
2256
2275
CTGTGTCTTCTTCAGAGGAA
50
362
|
|
1125543
N/A
N/A
2370
2389
TGTTGGTTGATGAAAATGTT
55
363
|
|
1125571
N/A
N/A
2482
2501
CTTGATTCAGGTGTGTGCTA
73
364
|
|
1125599
N/A
N/A
2628
2647
TCAGTCTACTTAGTGCAACG
98
365
|
|
1125627
N/A
N/A
2739
2758
TTTGTGGGCATGCACAGAGC
31
366
|
|
1125655
N/A
N/A
2835
2854
ATCCTCATTCTTCTCTGGTT
26
367
|
|
1125683
N/A
N/A
2942
2961
AAGTAGGCACTTAATAATAC
73
368
|
|
1125711
N/A
N/A
3045
3064
TCAGGAGTCAGATAGCTGGA
100
369
|
|
1125739
N/A
N/A
3439
3458
TCCCTCAGGGTCTGGTCAGG
25
370
|
|
1125767
N/A
N/A
3637
3656
GTCTCCCAGGCCCCTGCTCC
77
371
|
|
1125795
N/A
N/A
3749
3768
TAGAAAGAGACAAGGCTTCC
102
372
|
|
1125823
N/A
N/A
3892
3911
GGTGATACCAGGAGAGTAAT
93
373
|
|
1125851
N/A
N/A
4018
4037
TCCAAGCTTGGTTTACCCAC
38
374
|
|
1125879
N/A
N/A
4125
4144
TCTGAGAGATGGACATGGTG
69
375
|
|
1125907
N/A
N/A
4313
4332
TATCCAGCAACCTATTCTGT
152
376
|
|
1125935
N/A
N/A
4538
4557
CTCCTCCCTGGCCCGTTCCC
64
377
|
|
1125963
N/A
N/A
4635
4654
GGGAACGCAGTGAGCCACCC
97
378
|
|
1125991
N/A
N/A
4862
4881
AGGCTGGGCTCTCCTGCCTC
105
379
|
|
1126019
N/A
N/A
5386
5405
TCCTTCCCCCCCCCACTTCC
37
380
|
|
1126047
N/A
N/A
5754
5773
TGGGAGACGGAGGAGCCGCG
87
381
|
|
1126075
N/A
N/A
6161
6180
GCTAACCCGGGCGGAGAGGA
36
382
|
|
1126103
N/A
N/A
6376
6395
GCGGGCTTCTTCCGCCTAAC
66
383
|
|
1126131
N/A
N/A
6484
6503
CAGTGAATCCCAGGCCCTGG
34
384
|
|
1126159
N/A
N/A
6624
6643
CCAGAGTCGCAGAACCTGGC
34
385
|
|
1126187
N/A
N/A
6715
6734
TCAATTTCATAGGCAAGGAG
28
386
|
|
1126215
N/A
N/A
6842
6861
CTTCTTCACACCCCATCTGA
100
387
|
|
1126243
N/A
N/A
7109
7128
CCGGAATCACCCTGGGTCGG
28†
388
|
|
TABLE 6
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with full PS internucleoside linkages (Huh7,
|
electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
FXII
SEQ
|
Compound
Start
Stop
Start
Stop
(% UTC)
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
RTS40528
NO
|
|
413637
1748
1767
7136
7155
GCTTGGTCCTCACACACCAG
12†
80
|
|
462073
216
235
3567
3586
GGTACATTTGTGGTACAGCT
20
183
|
|
1124193
80
99
80
99
GTTGACTCCAAGCTCACCAG
34
389
|
|
1124221
138
157
505
524
GTGCTCTTCAGCTTTGTACT
37
390
|
|
1124247
174
193
3525
3544
GTGGCAGGGCTCCCCGGTGA
54
391
|
|
1124270
228
247
3579
3598
CCGGCCCTTGTGGGTACATT
51
392
|
|
1124298
293
312
3800
3819
CAGTATCCCCATCGCTGGTC
36
393
|
|
1124326
333
352
N/A
N/A
GTGTTTGCTGCAGTGGTCTT
56
394
|
|
1124354
406
425
4214
4233
TGAGGTGTTGTGGACAGAGA
56
395
|
|
1124382
449
468
4394
4413
AGCTGAGGCTCAAAGCACTT
38
396
|
|
1124409
494
513
4439
4458
TGCTCAGTTCTATACCATAT
31
397
|
|
1124437
535
554
4480
4499
GGGCATCAGGACCCTTGCAC
66
398
|
|
1124464
569
588
N/A
N/A
GTGCGGCAGGCCTGGCTGGC
70
399
|
|
1124491
609
628
4693
4712
CTCCACCTCTAGGCAGCGAC
28
400
|
|
1124519
655
674
4739
4758
AGAAGGCTCCGGTGTAGCCC
102
401
|
|
1124547
711
730
4940
4959
GCCGCGGTAGCTGAGCCCGC
58
402
|
|
1124574
765
784
4994
5013
GGCCTCCGAGGCCCACGGCT
44
403
|
|
1124597
843
862
N/A
N/A
GTTGTCCGGGTTCCGGCAGA
45
404
|
|
1124622
904
923
5218
5237
GGTCGCAGTACTCCCAGCTC
42
405
|
|
1124650
996
1015
5310
5329
CGGTGCCGGCTGCGCGGGCA
90
406
|
|
1124678
1073
1092
5492
5511
GGCTGCTCCCGCTTCGCCGG
92
407
|
|
1124706
1118
1137
5537
5556
AGCCGCTGCCCGCAGCTCAG
39
408
|
|
1124732
1155
1174
5574
5593
GCCAACGACGCGGGTCATCG
38
409
|
|
1124759
1227
1246
5646
5665
GCCGGCGCAGAAACTGTGGC
55
410
|
|
1124786
1313
1332
5973
5992
CCGAGCACCACCGTCAGATC
40
411
|
|
1124814
1399
1418
6059
6078
TGACGGGCGAGAAGGCCTCG
59
412
|
|
1124841
1462
1481
6205
6224
GGAGCGCGCAGCTGCCGTCC
44
413
|
|
1124865
1550
1569
6293
6312
TGGCCCCAGCCGGCCACCTG
53
414
|
|
1124892
1581
1600
6879
6898
GCTGGCATATTCCTCCGCCC
78
415
|
|
1124919
1622
1641
6920
6939
CGCTCCAGGGAGAGGAACGG
80
416
|
|
1124972
1795
1814
7183
7202
CCGATCCCCAGCTGATGATG
41
417
|
|
1124997
1842
1861
7230
7249
GTAGGCCACATCGGTGTAGA
39
418
|
|
1125023
1873
1892
7261
7280
AAACGGTGTGCTCCCGGATC
26
419
|
|
1125049
1917
1936
7305
7324
GCGGAATCACCAAGGAGGGA
20
420
|
|
1125076
1952
1971
7340
7359
AATCTTGCCTTCCATGCCCC
22
421
|
|
1125102
2007
2026
7395
7414
TTGAGTTCCTGCGCCATCCT
15
422
|
|
1125124
N/A
N/A
100
119
CCACAGCACTCACCGAAAGT
102
423
|
|
1125152
N/A
N/A
184
203
CTGGACCCACAGGTCATGAG
87
424
|
|
1125180
N/A
N/A
280
299
AGGCTGTGCCCTTGTATCCA
109
425
|
|
1125208
N/A
N/A
374
393
TCACAGCCCAGAAATGCAGA
82
426
|
|
1125236
N/A
N/A
463
482
AGGTGGAATCTACAAGGGAG
100
427
|
|
1125264
N/A
N/A
609
628
CAGGTAGGCACTAGACTAGA
81
428
|
|
1125292
N/A
N/A
696
715
TGGGCACTTAGACACAGCCC
52
429
|
|
1125320
N/A
N/A
865
884
TTGCCCATTGAAGGCCCTGG
39
430
|
|
1125348
N/A
N/A
963
982
GAGAACTCAAATCCCTCGCC
50
431
|
|
1125376
N/A
N/A
1070
1089
ATGCACCAGGTGATGGGCTG
62
432
|
|
1125404
N/A
N/A
1203
1222
GTAGGTAATTTAGTGTCTGG
59
433
|
|
1125432
N/A
N/A
1581
1600
ACTCAACCCCACCAAGTCAG
52
434
|
|
1125460
N/A
N/A
1753
1772
ATGGGATGGACGGACAGAGA
58
435
|
|
1125488
N/A
N/A
2129
2148
TTGCCAAGGACCAAACCAGA
72
436
|
|
1125516
N/A
N/A
2259
2278
TGCCTGTGTCTTCTTCAGAG
61
437
|
|
1125544
N/A
N/A
2373
2392
TGCTGTTGGTTGATGAAAAT
62
438
|
|
1125572
N/A
N/A
2485
2504
ACACTTGATTCAGGTGTGTG
74
439
|
|
1125600
N/A
N/A
2634
2653
TCCTTCTCAGTCTACTTAGT
49
440
|
|
1125628
N/A
N/A
2742
2761
CCATTTGTGGGCATGCACAG
75
441
|
|
1125656
N/A
N/A
2838
2857
TTGATCCTCATTCTTCTCTG
77
442
|
|
1125684
N/A
N/A
2950
2969
TTGCCACAAAGTAGGCACTT
51
443
|
|
1125712
N/A
N/A
3048
3067
GGTTCAGGAGTCAGATAGCT
66
444
|
|
1125740
N/A
N/A
3442
3461
CTGTCCCTCAGGGTCTGGTC
61
445
|
|
1125768
N/A
N/A
3640
3659
CATGTCTCCCAGGCCCCTGC
73
446
|
|
1125796
N/A
N/A
3752
3771
CTGTAGAAAGAGACAAGGCT
59
447
|
|
1125824
N/A
N/A
3895
3914
TCTGGTGATACCAGGAGAGT
48
448
|
|
1125852
N/A
N/A
4021
4040
GTTTCCAAGCTTGGTTTACC
54
449
|
|
1125880
N/A
N/A
4128
4147
TGGTCTGAGAGATGGACATG
59
450
|
|
1125908
N/A
N/A
4316
4335
GAGTATCCAGCAACCTATTC
67
451
|
|
1125936
N/A
N/A
4541
4560
ACGCTCCTCCCTGGCCCGTT
60
452
|
|
1125964
N/A
N/A
4638
4657
GGAGGGAACGCAGTGAGCCA
127
453
|
|
1125992
N/A
N/A
4866
4885
GCCAAGGCTGGGCTCTCCTG
57
454
|
|
1126020
N/A
N/A
5389
5408
TCCTCCTTCCCCCCCCCACT
82
455
|
|
1126048
N/A
N/A
5757
5776
CGCTGGGAGACGGAGGAGCC
116
456
|
|
1126076
N/A
N/A
6164
6183
AGAGCTAACCCGGGCGGAGA
120
457
|
|
1126104
N/A
N/A
6379
6398
GTCGCGGGCTTCTTCCGCCT
54
458
|
|
1126132
N/A
N/A
6487
6506
CAGCAGTGAATCCCAGGCCC
96
459
|
|
1126160
N/A
N/A
6627
6646
CATCCAGAGTCGCAGAACCT
105
460
|
|
1126188
N/A
N/A
6718
6737
AATTCAATTTCATAGGCAAG
73
461
|
|
1126216
N/A
N/A
6845
6864
CGCCTTCTTCACACCCCATC
68
462
|
|
1126244
N/A
N/A
7112
7131
CCTCCGGAATCACCCTGGGT
31†
463
|
|
TABLE 7
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with full PS internucleoside linkages (Huh7,
|
electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
FXII
SEQ
|
Compound
Start
Stop
Start
Stop
(% UTC)
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
RTS40528
NO
|
|
462073
216
235
3567
3586
GGTACATTTGTGGTACAGCT
15
183
|
|
564842
1463
1482
6206
6225
AGGAGCGCGCAGCTGCCGTC
44
464
|
|
1124194
82
101
82
101
GTGTTGACTCCAAGCTCACC
63
465
|
|
1124222
139
158
506
525
TGTGCTCTTCAGCTTTGTAC
25
466
|
|
1124248
175
194
3526
3545
AGTGGCAGGGCTCCCCGGTG
48
467
|
|
1124271
229
248
3580
3599
GCCGGCCCTTGTGGGTACAT
49
468
|
|
1124299
294
313
3801
3820
ACAGTATCCCCATCGCTGGT
46
469
|
|
1124327
334
353
4142
4161
TGTGTTTGCTGCAGTGGTCT
52
470
|
|
1124355
407
426
4215
4234
GTGAGGTGTTGTGGACAGAG
110
471
|
|
1124383
450
469
4395
4414
AAGCTGAGGCTCAAAGCACT
55
472
|
|
1124410
495
514
4440
4459
TTGCTCAGTTCTATACCATA
36
473
|
|
1124438
536
555
4481
4500
TGGGCATCAGGACCCTTGCA
51
474
|
|
1124465
570
589
N/A
N/A
GGTGCGGCAGGCCTGGCTGG
140
475
|
|
1124492
610
629
4694
4713
CCTCCACCTCTAGGCAGCGA
38
476
|
|
1124520
656
675
4740
4759
CAGAAGGCTCCGGTGTAGCC
63
477
|
|
1124548
712
731
4941
4960
GGCCGCGGTAGCTGAGCCCG
66
478
|
|
1124575
766
785
4995
5014
TGGCCTCCGAGGCCCACGGC
65
479
|
|
1124598
844
863
N/A
N/A
CGTTGTCCGGGTTCCGGCAG
33
480
|
|
1124623
905
924
5219
5238
AGGTCGCAGTACTCCCAGCT
47
481
|
|
1124651
997
1016
5311
5330
GCGGTGCCGGCTGCGCGGGC
57
482
|
|
1124679
1075
1094
5494
5513
GCGGCTGCTCCCGCTTCGCC
45
483
|
|
1124707
1120
1139
5539
5558
GGAGCCGCTGCCCGCAGCTC
47
484
|
|
1124733
1156
1175
5575
5594
CGCCAACGACGCGGGTCATC
42
485
|
|
1124760
1228
1247
5647
5666
TGCCGGCGCAGAAACTGTGG
64
486
|
|
1124787
1314
1333
5974
5993
GCCGAGCACCACCGTCAGAT
36
487
|
|
1124815
1400
1419
6060
6079
CTGACGGGCGAGAAGGCCTC
45
488
|
|
1124866
1551
1570
6294
6313
GTGGCCCCAGCCGGCCACCT
118
489
|
|
1124893
1582
1601
6880
6899
AGCTGGCATATTCCTCCGCC
81
490
|
|
1124920
1623
1642
6921
6940
GCGCTCCAGGGAGAGGAACG
79
491
|
|
1124945
1749
1768
7137
7156
AGCTTGGTCCTCACACACCA
121
492
|
|
1124973
1796
1815
7184
7203
CCCGATCCCCAGCTGATGAT
39
493
|
|
1124998
1843
1862
7231
7250
AGTAGGCCACATCGGTGTAG
31
494
|
|
1125024
1874
1893
7262
7281
GAAACGGTGTGCTCCCGGAT
34
495
|
|
1125050
1918
1937
7306
7325
TGCGGAATCACCAAGGAGGG
23
496
|
|
1125077
1954
1973
7342
7361
ACAATCTTGCCTTCCATGCC
23
497
|
|
1125103
2009
2028
7397
7416
TATTGAGTTCCTGCGCCATC
25
498
|
|
1125125
N/A
N/A
103
122
TTCCCACAGCACTCACCGAA
84
499
|
|
1125153
N/A
N/A
187
206
CACCTGGACCCACAGGTCAT
110
500
|
|
1125181
N/A
N/A
283
302
TGCAGGCTGTGCCCTTGTAT
78
501
|
|
1125209
N/A
N/A
377
396
CTCTCACAGCCCAGAAATGC
68
502
|
|
1125237
N/A
N/A
466
485
CCAAGGTGGAATCTACAAGG
78
503
|
|
1125265
N/A
N/A
612
631
CACCAGGTAGGCACTAGACT
64
504
|
|
1125293
N/A
N/A
716
735
CCTGCTCCGAGCCAGGCTCG
74
505
|
|
1125321
N/A
N/A
868
887
TCCTTGCCCATTGAAGGCCC
65
506
|
|
1125349
N/A
N/A
966
985
TTGGAGAACTCAAATCCCTC
66
507
|
|
1125377
N/A
N/A
1073
1092
TCCATGCACCAGGTGATGGG
58
508
|
|
1125405
N/A
N/A
1206
1225
CATGTAGGTAATTTAGTGTC
90
509
|
|
1125433
N/A
N/A
1584
1603
GAGACTCAACCCCACCAAGT
88
510
|
|
1125461
N/A
N/A
1756
1775
CAGATGGGATGGACGGACAG
65
511
|
|
1125489
N/A
N/A
2132
2151
CACTTGCCAAGGACCAAACC
86
512
|
|
1125517
N/A
N/A
2262
2281
AATTGCCTGTGTCTTCTTCA
57
513
|
|
1125545
N/A
N/A
2376
2395
ATATGCTGTTGGTTGATGAA
55
514
|
|
1125573
N/A
N/A
2488
2507
TAGACACTTGATTCAGGTGT
57
515
|
|
1125601
N/A
N/A
2637
2656
GTGTCCTTCTCAGTCTACTT
68
516
|
|
1125629
N/A
N/A
2745
2764
TGGCCATTTGTGGGCATGCA
54
517
|
|
1125657
N/A
N/A
2841
2860
CAGTTGATCCTCATTCTTCT
59
518
|
|
1125685
N/A
N/A
2953
2972
CACTTGCCACAAAGTAGGCA
51
519
|
|
1125713
N/A
N/A
3051
3070
TTGGGTTCAGGAGTCAGATA
116
520
|
|
1125741
N/A
N/A
3445
3464
GCACTGTCCCTCAGGGTCTG
68
521
|
|
1125769
N/A
N/A
3643
3662
GTACATGTCTCCCAGGCCCC
57
522
|
|
1125797
N/A
N/A
3755
3774
CACCTGTAGAAAGAGACAAG
148
523
|
|
1125825
N/A
N/A
3898
3917
GGGTCTGGTGATACCAGGAG
56
524
|
|
1125853
N/A
N/A
4024
4043
CAAGTTTCCAAGCTTGGTTT
53
525
|
|
1125881
N/A
N/A
4131
4150
CAGTGGTCTGAGAGATGGAC
56
526
|
|
1125909
N/A
N/A
4319
4338
TCCGAGTATCCAGCAACCTA
88
527
|
|
1125937
N/A
N/A
4557
4576
CCAGCCTGTCTTCCTGACGC
63
528
|
|
1125965
N/A
N/A
4657
4676
GTTGGTGCGGCAGGCTTGGG
60
529
|
|
1125993
N/A
N/A
4869
4888
GCAGCCAAGGCTGGGCTCTC
75
530
|
|
1126021
N/A
N/A
5392
5411
GGCTCCTCCTTCCCCCCCCC
54
531
|
|
1126049
N/A
N/A
5760
5779
CTGCGCTGGGAGACGGAGGA
181
532
|
|
1126077
N/A
N/A
6167
6186
AACAGAGCTAACCCGGGCGG
67
533
|
|
1126105
N/A
N/A
6382
6401
AAAGTCGCGGGCTTCTTCCG
59
534
|
|
1126133
N/A
N/A
6490
6509
TCCCAGCAGTGAATCCCAGG
88
535
|
|
1126161
N/A
N/A
6630
6649
ACCCATCCAGAGTCGCAGAA
72
536
|
|
1126189
N/A
N/A
6721
6740
ATTAATTCAATTTCATAGGC
76
537
|
|
1126217
N/A
N/A
6862
6881
CCCCTGCGAACACAGAGCGC
105
538
|
|
TABLE 8
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with full PS internucleoside linkages (Huh7,
|
electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
FXII
SEQ
|
Compound
Start
Stop
Start
Stop
(% UTC)
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
RTS40528
NO
|
|
413644
1844
1863
7232
7251
TAGTAGGCCACATCGGTGTA
32
87
|
|
462073
216
235
3567
3586
GGTACATTTGTGGTACAGCT
48
183
|
|
564822
862
881
5176
5195
AGCACCACGGGCGGATGTCG
69
539
|
|
1124167
36
55
36
55
CCTCATGGCATCCGTCCGTT
42
540
|
|
1124195
83
102
83
102
AGTGTTGACTCCAAGCTCAC
45
541
|
|
1124223
141
160
508
527
TGTGTGCTCTTCAGCTTTGT
32
542
|
|
1124249
177
196
3528
3547
GAAGTGGCAGGGCTCCCCGG
54
543
|
|
1124272
231
250
3582
3601
TGGCCGGCCCTTGTGGGTAC
141
544
|
|
1124300
296
315
3803
3822
AAACAGTATCCCCATCGCTG
66
545
|
|
1124328
335
354
4143
4162
CTGTGTTTGCTGCAGTGGTC
38
546
|
|
1124356
408
427
4216
4235
AGTGAGGTGTTGTGGACAGA
73
547
|
|
1124384
451
470
4396
4415
GAAGCTGAGGCTCAAAGCAC
32
548
|
|
1124411
496
515
4441
4460
CTTGCTCAGTTCTATACCAT
52
549
|
|
1124439
537
556
4482
4501
GTGGGCATCAGGACCCTTGC
45
550
|
|
1124466
571
590
N/A
N/A
TGGTGCGGCAGGCCTGGCTG
146
551
|
|
1124493
612
631
4696
4715
GCCCTCCACCTCTAGGCAGC
70
552
|
|
1124521
657
676
4741
4760
GCAGAAGGCTCCGGTGTAGC
97
553
|
|
1124549
714
733
4943
4962
CAGGCCGCGGTAGCTGAGCC
99
554
|
|
1124576
767
786
4996
5015
GTGGCCTCCGAGGCCCACGG
78
555
|
|
1124624
907
926
5221
5240
CCAGGTCGCAGTACTCCCAG
51
556
|
|
1124652
998
1017
5312
5331
GGCGGTGCCGGCTGCGCGGG
70
557
|
|
1124680
1076
1095
5495
5514
GGCGGCTGCTCCCGCTTCGC
67
558
|
|
1124708
1121
1140
5540
5559
CGGAGCCGCTGCCCGCAGCT
136
559
|
|
1124734
1158
1177
5577
5596
CCCGCCAACGACGCGGGTCA
51
560
|
|
1124761
1229
1248
5648
5667
CTGCCGGCGCAGAAACTGTG
54
561
|
|
1124788
1315
1334
5975
5994
GGCCGAGCACCACCGTCAGA
100
562
|
|
1124816
1401
1420
6061
6080
GCTGACGGGCGAGAAGGCCT
70
563
|
|
1124842
1464
1483
6207
6226
CAGGAGCGCGCAGCTGCCGT
40
564
|
|
1124867
1552
1571
6295
6314
GGTGGCCCCAGCCGGCCACC
66
565
|
|
1124894
1583
1602
6881
6900
AAGCTGGCATATTCCTCCGC
80
566
|
|
1124921
1657
1676
6955
6974
GGATGGAGGATCCGTGCACG
30†
567
|
|
1124946
1750
1769
7138
7157
CAGCTTGGTCCTCACACACC
18†
568
|
|
1124974
1797
1816
7185
7204
GCCCGATCCCCAGCTGATGA
43
569
|
|
1125025
1876
1895
7264
7283
AGGAAACGGTGTGCTCCCGG
42
570
|
|
1125051
1919
1938
7307
7326
CTGCGGAATCACCAAGGAGG
26
571
|
|
1125078
1955
1974
7343
7362
CACAATCTTGCCTTCCATGC
37
572
|
|
1125104
2010
2029
7398
7417
TTATTGAGTTCCTGCGCCAT
22
573
|
|
1125126
N/A
N/A
106
125
TGGTTCCCACAGCACTCACC
73
574
|
|
1125154
N/A
N/A
190
209
AGTCACCTGGACCCACAGGT
78
575
|
|
1125182
N/A
N/A
286
305
AAATGCAGGCTGTGCCCTTG
69
576
|
|
1125210
N/A
N/A
380
399
TTCCTCTCACAGCCCAGAAA
60
577
|
|
1125238
N/A
N/A
469
488
TTCCCAAGGTGGAATCTACA
84
578
|
|
1125266
N/A
N/A
615
634
TAGCACCAGGTAGGCACTAG
66
579
|
|
1125294
N/A
N/A
721
740
AAGCACCTGCTCCGAGCCAG
56
580
|
|
1125322
N/A
N/A
871
890
CCTTCCTTGCCCATTGAAGG
60
581
|
|
1125350
N/A
N/A
969
988
AGCTTGGAGAACTCAAATCC
62
582
|
|
1125378
N/A
N/A
1076
1095
ATTTCCATGCACCAGGTGAT
62
583
|
|
1125406
N/A
N/A
1209
1228
TGGCATGTAGGTAATTTAGT
63
584
|
|
1125434
N/A
N/A
1587
1606
TTAGAGACTCAACCCCACCA
143
585
|
|
1125462
N/A
N/A
1759
1778
ATGCAGATGGGATGGACGGA
167
586
|
|
1125490
N/A
N/A
2135
2154
GTGCACTTGCCAAGGACCAA
73
587
|
|
1125518
N/A
N/A
2265
2284
GAGAATTGCCTGTGTCTTCT
55
588
|
|
1125546
N/A
N/A
2379
2398
ATTATATGCTGTTGGTTGAT
123
589
|
|
1125574
N/A
N/A
2513
2532
GCTACCTTAGGGAGAAAGCG
121
590
|
|
1125602
N/A
N/A
2640
2659
TGAGTGTCCTTCTCAGTCTA
69
591
|
|
1125630
N/A
N/A
2748
2767
TCATGGCCATTTGTGGGCAT
89
592
|
|
1125658
N/A
N/A
2845
2864
AGTACAGTTGATCCTCATTC
162
593
|
|
1125686
N/A
N/A
2956
2975
GAGCACTTGCCACAAAGTAG
91
594
|
|
1125714
N/A
N/A
3054
3073
AACTTGGGTTCAGGAGTCAG
105
595
|
|
1125742
N/A
N/A
3448
3467
CAGGCACTGTCCCTCAGGGT
141
596
|
|
1125770
N/A
N/A
3649
3668
GGCAGGGTACATGTCTCCCA
68
597
|
|
1125798
N/A
N/A
3758
3777
GCACACCTGTAGAAAGAGAC
82
598
|
|
1125826
N/A
N/A
3916
3935
CAGAATCCCAGGTGTGTGGG
54
599
|
|
1125854
N/A
N/A
4027
4046
CTCCAAGTTTCCAAGCTTGG
116
600
|
|
1125882
N/A
N/A
4134
4153
CTGCAGTGGTCTGAGAGATG
102
601
|
|
1125910
N/A
N/A
4325
4344
CAAGTCTCCGAGTATCCAGC
96
602
|
|
1125938
N/A
N/A
4560
4579
CTGCCAGCCTGTCTTCCTGA
80
603
|
|
1125966
N/A
N/A
4660
4679
CGGGTTGGTGCGGCAGGCTT
95
604
|
|
1125994
N/A
N/A
4875
4894
CCCTGGGCAGCCAAGGCTGG
67
605
|
|
1126022
N/A
N/A
5395
5414
CTCGGCTCCTCCTTCCCCCC
61
606
|
|
1126050
N/A
N/A
5763
5782
AAGCTGCGCTGGGAGACGGA
123
607
|
|
1126078
N/A
N/A
6170
6189
CGCAACAGAGCTAACCCGGG
76
608
|
|
1126106
N/A
N/A
6385
6404
ACCAAAGTCGCGGGCTTCTT
150
609
|
|
1126134
N/A
N/A
6493
6512
GGATCCCAGCAGTGAATCCC
70
610
|
|
1126162
N/A
N/A
6633
6652
ACCACCCATCCAGAGTCGCA
60
611
|
|
1126190
N/A
N/A
6724
6743
GCCATTAATTCAATTTCATA
72
612
|
|
1126218
N/A
N/A
6865
6884
CCGCCCCTGCGAACACAGAG
66
613
|
|
TABLE 9
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers
|
with full PS internucleoside linkages (Huh7,
|
oligofectamine, 200 nM modified
|
oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
Com-
3
3
4
4
SEQ
|
pound
Start
Stop
Start
Stop
Sequence
FXII
ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
462200
10845
10864
N/A
N/A
CTTCTCAGCA
63
614
|
TTTTCAAAGC
|
|
462201
10848
10867
N/A
N/A
TTCCTTCTCA
63
615
|
GCATTTTCAA
|
|
TABLE 10
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers
|
with full PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM
|
modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
Com-
3
3
4
4
SEQ
|
pound
Start
Stop
Start
Stop
Sequence
FXII
ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1126246
10849
10868
N/A
N/A
TTTCCTTCTC
130
616
|
AGCATTTTCA
|
|
TABLE 11
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers
|
with full PS internucleoside linkages
|
electroporation, 5000 nM modified
|
oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
Com-
3
3
4
4
SEQ
|
pound
Start
Stop
Start
Stop
Sequence
FXII
ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1124165
3458
3477
34
53
TCATGGCATC
60
617
|
CGTCCGTTGG
|
|
1126245
10846
10865
N/A
N/A
CCTTCTCAGC
77
618
|
ATTTTCAAAG
|
|
1124166
3459
3478
35
54
CTCATGGCAT
52
619
|
CCGTCCGTTG
|
|
Example 2: Effects of 5-10-5 MOE Gapmers on Human FXII RNA In Vitro, Single Dose
Modified oligonucleotides complementary to a FXII nucleic acid were synthesized and tested for their effect on FXII RNA levels in vitro. The modified oligonucleotides were tested in a series of experiments using similar culture conditions. The results for each separate experiment are presented in separate tables below.
The modified oligonucleotides in the tables below 5-10-5 MOE gapmers (i.e., they have a central region of ten 2′-deoxynucleosides flanked on each side by wings, each comprising five 2′-MOE nucleosides). The motif for the gapmers is (from 5′ to 3′): eeeeeddddddddddeeeee; wherein “d” represents a 2′-β-D-deoxyribosyl sugar moiety, and ‘e’ represents a 2′-MOE sugar moiety. The internucleoside linkage motif for the gapmers is (from 5′ to 3′): soooossssssssssooss; wherein ‘o’ represents a phosphodiester internucleoside linkage and ‘s’ represents a phosphorothioate internucleoside linkage. Each cytosine residue is a 5-methyl cytosine.
“Start site” indicates the 5′-most nucleoside of the target sequence to which the modified oligonucleotide is complementary. “Stop site” indicates the 3′-most nucleoside of the target sequence to which the modified oligonucleotide is complementary. As shown in the tables below, the modified oligonucleotides are 100% complementary to one or more of human FXII target sequences including the human FXII mRNA sequence designated herein as SEQ ID NO: 1 (ENSEMBL ID ENST00000253496.3 from ENSEMBL version 99: January 2020), the human FXII genomic sequence, designated herein as SEQ ID NO: 2 (ENSEMBL ID ENSG00000131187.9 from ENSEMBL version 99: January 2020), the human FXII genomic sequence, designated herein as SEQ ID No. 3 (the complement of GENBANK Accession No. NC_000005.10 truncated from truncated from nucleotides 177399001 to 177413000), and the human FXII mRNA sequence designated herein as SEQ ID No: 4 (GENBANK Accession No. NM 000505.3). ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular target sequence with 100% complementarity.
Cultured Huh7 cells, at a density of 20,000 cells per well, were transfected using electroporation with either 5000 nM or 2000 nM modified oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and FXII RNA levels were measured by quantitative real-time RTPCR. Human primer probe set RTS40528 (described herein above) was used to measure RNA levels. FXII RNA levels were normalized to total RNA content, as measured by RIBOGREEN®. Reduction of FXII RNA is presented in the tables below as percent FXII RNA relative to the amount in untreated control cells (% UTC). The symbol “†” indicates that the modified oligonucleotide is complementary to the target transcript within the amplicon region of the primer probe set. In such instances, additional assays using alternative primer probes must be performed to accurately assess the potency and efficacy of such modified oligonucleotides.
TABLE 12
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with
|
mixed PO/PS internucleoside linkages (Huh7,
|
electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130742
2
21
2
21
TCCAGGAGTCCAGATCAATA
67
620
|
|
1130770
37
56
37
56
CCCTCATGGCATCCGTCCGT
68
621
|
|
1130798
84
103
84
103
AAGTGTTGACTCCAAGCTCA
90
622
|
|
1130826
142
161
509
528
CTGTGTGCTCTTCAGCTTTG
13
623
|
|
1130854
178
197
3529
3548
GGAAGTGGCAGGGCTCCCCG
47
624
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
18
183
|
|
1130883
232
251
3583
3602
CTGGCCGGCCCTTGTGGGTA
21
625
|
|
1130911
297
316
3804
3823
CAAACAGTATCCCCATCGCT
28
626
|
|
1130939
338
357
4146
4165
GGGCTGTGTTTGCTGCAGTG
22
627
|
|
1130967
410
429
4218
4237
CCAGTGAGGTGTTGTGGACA
68
628
|
|
1130995
453
472
4398
4417
GAGAAGCTGAGGCTCAAAGC
38
629
|
|
1131023
498
517
4443
4462
TGCTTGCTCAGTTCTATACC
20
630
|
|
1131051
538
557
4483
4502
AGTGGGCATCAGGACCCTTG
93
277
|
|
1131079
573
592
N/A
N/A
GTTGGTGCGGCAGGCCTGGC
37
631
|
|
1131107
613
632
4697
4716
GGCCCTCCACCTCTAGGCAG
28
632
|
|
1131135
658
677
4742
4761
CGCAGAAGGCTCCGGTGTAG
54
633
|
|
1131163
715
734
4944
4963
CCAGGCCGCGGTAGCTGAGC
40
634
|
|
1131191
768
787
4997
5016
GGTGGCCTCCGAGGCCCACG
83
635
|
|
1131219
864
883
5178
5197
GAAGCACCACGGGCGGATGT
44
636
|
|
1131247
908
927
5222
5241
GCCAGGTCGCAGTACTCCCA
19
637
|
|
1131275
999
1018
5313
5332
CGGCGGTGCCGGCTGCGCGG
70
638
|
|
1131303
1077
1096
5496
5515
AGGCGGCTGCTCCCGCTTCG
38
639
|
|
1131331
1123
1142
5542
5561
TGCGGAGCCGCTGCCCGCAG
125
640
|
|
1131359
1159
1178
5578
5597
GCCCGCCAACGACGCGGGTC
56
641
|
|
1131387
1230
1249
5649
5668
GCTGCCGGCGCAGAAACTGT
57
642
|
|
1131415
1316
1335
5976
5995
TGGCCGAGCACCACCGTCAG
51
643
|
|
1131443
1402
1421
6062
6081
AGCTGACGGGCGAGAAGGCC
36
644
|
|
1131471
1465
1484
6208
6227
ACAGGAGCGCGCAGCTGCCG
30
645
|
|
1131499
1553
1572
6296
6315
TGGTGGCCCCAGCCGGCCAC
61
646
|
|
1131527
1585
1604
6883
6902
GGAAGCTGGCATATTCCTCC
57
647
|
|
1131555
1660
1679
6958
6977
GGAGGATGGAGGATCCGTGC
7†
648
|
|
1131583
1751
1770
7139
7158
GCAGCTTGGTCCTCACACAC
8†
649
|
|
1131611
1798
1817
7186
7205
AGCCCGATCCCCAGCTGATG
33
650
|
|
1131639
1845
1864
7233
7252
GTAGTAGGCCACATCGGTGT
62
651
|
|
1131667
1877
1896
7265
7284
CAGGAAACGGTGTGCTCCCG
28
652
|
|
1131695
1921
1940
7309
7328
CACTGCGGAATCACCAAGGA
13
653
|
|
1131723
1956
1975
7344
7363
ACACAATCTTGCCTTCCATG
26
654
|
|
1131751
2012
2031
7400
7419
CTTTATTGAGTTCCTGCGCC
20
95
|
|
1131779
N/A
N/A
109
128
TCCTGGTTCCCACAGCACTC
71
655
|
|
1131807
N/A
N/A
193
212
CCTAGTCACCTGGACCCACA
56
656
|
|
1131835
N/A
N/A
289
308
CAGAAATGCAGGCTGTGCCC
71
657
|
|
1131863
N/A
N/A
386
405
TGCAGCTTCCTCTCACAGCC
120
658
|
|
1131891
N/A
N/A
513
532
CCGACTGTGTGCTCTTCAGC
21
659
|
|
1131919
N/A
N/A
618
637
ACCTAGCACCAGGTAGGCAC
39
660
|
|
1131947
N/A
N/A
724
743
CTCAAGCACCTGCTCCGAGC
54
661
|
|
1131975
N/A
N/A
874
893
GATCCTTCCTTGCCCATTGA
122
662
|
|
1132003
N/A
N/A
972
991
TGAAGCTTGGAGAACTCAAA
76
663
|
|
1132031
N/A
N/A
1079
1098
AGCATTTCCATGCACCAGGT
70
664
|
|
1132059
N/A
N/A
1212
1231
GCCTGGCATGTAGGTAATTT
60
665
|
|
1132087
N/A
N/A
1590
1609
TGCTTAGAGACTCAACCCCA
67
666
|
|
1132115
N/A
N/A
1762
1781
AAGATGCAGATGGGATGGAC
79
667
|
|
1132143
N/A
N/A
2138
2157
AAAGTGCACTTGCCAAGGAC
60
668
|
|
1132171
N/A
N/A
2268
2287
AATGAGAATTGCCTGTGTCT
41
669
|
|
1132199
N/A
N/A
2382
2401
CAGATTATATGCTGTTGGTT
49
670
|
|
1132227
N/A
N/A
2516
2535
AAGGCTACCTTAGGGAGAAA
48
671
|
|
1132255
N/A
N/A
2643
2662
GAATGAGTGTCCTTCTCAGT
46
672
|
|
1132283
N/A
N/A
2751
2770
TTTTCATGGCCATTTGTGGG
60
673
|
|
1132311
N/A
N/A
2848
2867
AATAGTACAGTTGATCCTCA
46
674
|
|
1132339
N/A
N/A
2959
2978
GAAGAGCACTTGCCACAAAG
106
675
|
|
1132367
N/A
N/A
3060
3079
AAAAAAAACTTGGGTTCAGG
133
676
|
|
1132395
N/A
N/A
3451
3470
GCTCAGGCACTGTCCCTCAG
41
677
|
|
1132423
N/A
N/A
3652
3671
ACAGGCAGGGTACATGTCTC
45
678
|
|
1132451
N/A
N/A
3761
3780
GTAGCACACCTGTAGAAAGA
45
679
|
|
1132479
N/A
N/A
3919
3938
GTCCAGAATCCCAGGTGTGT
46
680
|
|
1132507
N/A
N/A
4030
4049
CTACTCCAAGTTTCCAAGCT
97
681
|
|
1132535
N/A
N/A
4137
4156
TTGCTGCAGTGGTCTGAGAG
36
682
|
|
1132563
N/A
N/A
4328
4347
TGCCAAGTCTCCGAGTATCC
56
683
|
|
1132591
N/A
N/A
4563
4582
CTCCTGCCAGCCTGTCTTCC
70
684
|
|
1132619
N/A
N/A
4763
4782
CAGACCCTCACTCACCCACG
67
685
|
|
1132647
N/A
N/A
4878
4897
GCTCCCTGGGCAGCCAAGGC
37
686
|
|
1132675
N/A
N/A
5398
5417
CCTCTCGGCTCCTCCTTCCC
78
687
|
|
1132703
N/A
N/A
5766
5785
TGGAAGCTGCGCTGGGAGAC
122
688
|
|
1132731
N/A
N/A
6173
6192
AGGCGCAACAGAGCTAACCC
32
689
|
|
1132759
N/A
N/A
6388
6407
GATACCAAAGTCGCGGGCTT
100
690
|
|
1132787
N/A
N/A
6512
6531
GTATCCCCAGGAGATTTGGG
66
691
|
|
1132815
N/A
N/A
6636
6655
CCCACCACCCATCCAGAGTC
56
692
|
|
1132843
N/A
N/A
6727
6746
GGTGCCATTAATTCAATTTC
109
693
|
|
1132871
N/A
N/A
6868
6887
CCTCCGCCCCTGCGAACACA
55
694
|
|
TABLE 13
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130743
3
22
3
22
ATCCAGGAGTCCAGATCAAT
48
695
|
|
1130771
38
57
38
57
GCCCTCATGGCATCCGTCCG
57
696
|
|
1130799
85
104
85
104
AAAGTGTTGACTCCAAGCTC
53
697
|
|
1130827
144
163
511
530
GACTGTGTGCTCTTCAGCTT
32
698
|
|
1130855
197
216
3548
3567
TGCCGGTGGTACTGGAAGGG
53
699
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
13
183
|
|
1130884
233
252
3584
3603
CCTGGCCGGCCCTTGTGGGT
50
700
|
|
1130912
298
317
3805
3824
CCAAACAGTATCCCCATCGC
25
701
|
|
1130940
357
376
4165
4184
GGTCCCTCCTTTCTGGCAGG
76
702
|
|
1130968
411
430
4219
4238
TCCAGTGAGGTGTTGTGGAC
42
703
|
|
1130996
456
475
4401
4420
CCGGAGAAGCTGAGGCTCAA
22
704
|
|
1131024
499
518
4444
4463
CTGCTTGCTCAGTTCTATAC
14
705
|
|
1131052
539
558
4484
4503
CAGTGGGCATCAGGACCCTT
46
706
|
|
1131080
574
593
N/A
N/A
GGTTGGTGCGGCAGGCCTGG
22
707
|
|
1131108
615
634
4699
4718
GTGGCCCTCCACCTCTAGGC
62
708
|
|
1131136
660
679
4744
4763
GTCGCAGAAGGCTCCGGTGT
64
709
|
|
1131164
716
735
4945
4964
GCCAGGCCGCGGTAGCTGAG
46
710
|
|
1131192
772
791
5001
5020
GGTAGGTGGCCTCCGAGGCC
52
711
|
|
1131220
865
884
5179
5198
CGAAGCACCACGGGCGGATG
95
712
|
|
1131248
909
928
5223
5242
TGCCAGGTCGCAGTACTCCC
29
713
|
|
1131276
1000
1019
5314
5333
TCGGCGGTGCCGGCTGCGCG
39
714
|
|
1131304
1078
1097
5497
5516
AAGGCGGCTGCTCCCGCTTC
78
715
|
|
1131332
1124
1143
5543
5562
TTGCGGAGCCGCTGCCCGCA
65
716
|
|
1131360
1161
1180
5580
5599
CAGCCCGCCAACGACGCGGG
62
717
|
|
1131388
1232
1251
5651
5670
AGGCTGCCGGCGCAGAAACT
34
718
|
|
1131416
1317
1336
5977
5996
CTGGCCGAGCACCACCGTCA
13
719
|
|
1131444
1405
1424
6065
6084
GGTAGCTGACGGGCGAGAAG
48
720
|
|
1131472
1467
1486
6210
6229
CGACAGGAGCGCGCAGCTGC
59
721
|
|
1131500
1554
1573
6297
6316
CTGGTGGCCCCAGCCGGCCA
53
722
|
|
1131528
1586
1605
6884
6903
AGGAAGCTGGCATATTCCTC
47
723
|
|
1131556
1679
1698
6977
6996
CCTGCGCAGAGCATGCCGGG
71†
75
|
|
1131584
1752
1771
7140
7159
TGCAGCTTGGTCCTCACACA
17†
724
|
|
1131612
1799
1818
7187
7206
CAGCCCGATCCCCAGCTGAT
80
725
|
|
1131640
1846
1865
7234
7253
GGTAGTAGGCCACATCGGTG
19
726
|
|
1131668
1878
1897
7266
7285
TCAGGAAACGGTGTGCTCCC
46
727
|
|
1131696
1922
1941
7310
7329
TCACTGCGGAATCACCAAGG
21
728
|
|
1131724
1958
1977
7346
7365
GGACACAATCTTGCCTTCCA
7
729
|
|
1131752
2013
2032
7401
7420
ACTTTATTGAGTTCCTGCGC
11
162
|
|
1131780
N/A
N/A
112
131
CAATCCTGGTTCCCACAGCA
64
730
|
|
1131808
N/A
N/A
196
215
CCTCCTAGTCACCTGGACCC
60
731
|
|
1131836
N/A
N/A
292
311
CCCCAGAAATGCAGGCTGTG
114
732
|
|
1131864
N/A
N/A
389
408
GTCTGCAGCTTCCTCTCACA
56
733
|
|
1131892
N/A
N/A
516
535
TTACCGACTGTGTGCTCTTC
18
734
|
|
1131920
N/A
N/A
621
640
AAGACCTAGCACCAGGTAGG
58
735
|
|
1131948
N/A
N/A
727
746
TATCTCAAGCACCTGCTCCG
25
736
|
|
1131976
N/A
N/A
877
896
CTTGATCCTTCCTTGCCCAT
58
737
|
|
1132004
N/A
N/A
975
994
AATTGAAGCTTGGAGAACTC
99
738
|
|
1132032
N/A
N/A
1082
1101
TAAAGCATTTCCATGCACCA
93
739
|
|
1132060
N/A
N/A
1215
1234
CATGCCTGGCATGTAGGTAA
53
740
|
|
1132088
N/A
N/A
1593
1612
CCATGCTTAGAGACTCAACC
82
741
|
|
1132116
N/A
N/A
1765
1784
AAAAAGATGCAGATGGGATG
86
742
|
|
1132144
N/A
N/A
2141
2160
GGCAAAGTGCACTTGCCAAG
85
743
|
|
1132172
N/A
N/A
2271
2290
AATAATGAGAATTGCCTGTG
117
744
|
|
1132200
N/A
N/A
2385
2404
AACCAGATTATATGCTGTTG
41
745
|
|
1132228
N/A
N/A
2519
2538
AAGAAGGCTACCTTAGGGAG
40
746
|
|
1132256
N/A
N/A
2646
2665
ATTGAATGAGTGTCCTTCTC
50
747
|
|
1132284
N/A
N/A
2754
2773
ATGTTTTCATGGCCATTTGT
45
748
|
|
1132312
N/A
N/A
2851
2870
AATAATAGTACAGTTGATCC
62
749
|
|
1132340
N/A
N/A
2962
2981
CAGGAAGAGCACTTGCCACA
52
750
|
|
1132368
N/A
N/A
3370
3389
CTCTAAAAGTTGGGTTCAGG
51
751
|
|
1132396
N/A
N/A
3457
3476
TCAACTGCTCAGGCACTGTC
53
752
|
|
1132424
N/A
N/A
3655
3674
AGGACAGGCAGGGTACATGT
51
753
|
|
1132452
N/A
N/A
3764
3783
GTGGTAGCACACCTGTAGAA
28
754
|
|
1132480
N/A
N/A
3922
3941
TGGGTCCAGAATCCCAGGTG
59
755
|
|
1132508
N/A
N/A
4033
4052
TTGCTACTCCAAGTTTCCAA
50
756
|
|
1132536
N/A
N/A
4140
4159
TGTTTGCTGCAGTGGTCTGA
21
757
|
|
1132564
N/A
N/A
4334
4353
GGACCATGCCAAGTCTCCGA
41
758
|
|
1132592
N/A
N/A
4566
4585
GGCCTCCTGCCAGCCTGTCT
49
759
|
|
1132620
N/A
N/A
4766
4785
CCCCAGACCCTCACTCACCC
90
760
|
|
1132648
N/A
N/A
4899
4918
TGGTGTCTGAGGAGAAAGGG
71
761
|
|
1132676
N/A
N/A
5401
5420
CGCCCTCTCGGCTCCTCCTT
44
762
|
|
1132704
N/A
N/A
5841
5860
CTCCCCGGGAGCTCCGGAGG
123
763
|
|
1132732
N/A
N/A
6176
6195
TGAAGGCGCAACAGAGCTAA
21
764
|
|
1132760
N/A
N/A
6394
6413
CGGAACGATACCAAAGTCGC
85
765
|
|
1132788
N/A
N/A
6515
6534
CCTGTATCCCCAGGAGATTT
21
766
|
|
1132816
N/A
N/A
6639
6658
TCCCCCACCACCCATCCAGA
87
767
|
|
1132844
N/A
N/A
6730
6749
GGAGGTGCCATTAATTCAAT
46
768
|
|
1132872
N/A
N/A
6871
6890
ATTCCTCCGCCCCTGCGAAC
150
769
|
|
TABLE 14
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with
|
mixed PO/PS internucleoside linkages (Huh7,
|
electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
FXII
SEQ ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130800
89
108
N/A
N/A
ATCGAAAGTGTTGACTCCAA
28
770
|
|
1130828
145
164
512
531
CGACTGTGTGCTCTTCAGCT
14
771
|
|
1130856
199
218
3550
3569
GCTGCCGGTGGTACTGGAAG
22
772
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
21
183
|
|
1130885
234
253
3585
3604
GCCTGGCCGGCCCTTGTGGG
51
773
|
|
1130913
304
323
3811
3830
TGGGCTCCAAACAGTATCCC
22
774
|
|
1130941
359
378
4167
4186
CAGGTCCCTCCTTTCTGGCA
46
775
|
|
1130969
413
432
4221
4240
TTTCCAGTGAGGTGTTGTGG
25
776
|
|
1130997
457
476
4402
4421
ACCGGAGAAGCTGAGGCTCA
19
777
|
|
1131025
500
519
4445
4464
GCTGCTTGCTCAGTTCTATA
19
778
|
|
1131053
540
559
4485
4504
GCAGTGGGCATCAGGACCCT
22
779
|
|
1131081
576
595
N/A
N/A
CGGGTTGGTGCGGCAGGCCT
65
780
|
|
1131109
616
635
4700
4719
GGTGGCCCTCCACCTCTAGG
38
781
|
|
1131137
661
680
4745
4764
CGTCGCAGAAGGCTCCGGTG
94
782
|
|
1131165
717
736
4946
4965
GGCCAGGCCGCGGTAGCTGA
35
783
|
|
1131193
773
792
5002
5021
CGGTAGGTGGCCTCCGAGGC
30
784
|
|
1131221
867
886
5181
5200
CACGAAGCACCACGGGCGGA
38
785
|
|
1131249
911
930
5225
5244
TGTGCCAGGTCGCAGTACTC
24
786
|
|
1131277
1001
1020
5315
5334
TTCGGCGGTGCCGGCTGCGC
25
787
|
|
1131305
1079
1098
5498
5517
GAAGGCGGCTGCTCCCGCTT
39
788
|
|
1131333
1126
1145
5545
5564
TCTTGCGGAGCCGCTGCCCG
34
789
|
|
1131361
1162
1181
5581
5600
CCAGCCCGCCAACGACGCGG
31
790
|
|
1131389
1233
1252
5652
5671
GAGGCTGCCGGCGCAGAAAC
39
791
|
|
1131417
1318
1337
5978
5997
CCTGGCCGAGCACCACCGTC
26
792
|
|
1131445
1408
1427
6068
6087
GCTGGTAGCTGACGGGCGAG
26
793
|
|
1131473
1468
1487
6211
6230
GCGACAGGAGCGCGCAGCTG
21
794
|
|
1131501
1555
1574
6298
6317
ACTGGTGGCCCCAGCCGGCC
51
795
|
|
1131529
1587
1606
6885
6904
CAGGAAGCTGGCATATTCCT
82
796
|
|
1131557
1680
1699
6978
6997
CCCTGCGCAGAGCATGCCGG
28†
797
|
|
1131585
1753
1772
7141
7160
CTGCAGCTTGGTCCTCACAC
6†
798
|
|
1131613
1800
1819
7188
7207
ACAGCCCGATCCCCAGCTGA
39
799
|
|
1131641
1847
1866
7235
7254
AGGTAGTAGGCCACATCGGT
19
800
|
|
1131669
1879
1898
7267
7286
ATCAGGAAACGGTGTGCTCC
24
801
|
|
1131697
1923
1942
7311
7330
CTCACTGCGGAATCACCAAG
8
802
|
|
1131725
1960
1979
7348
7367
TGGGACACAATCTTGCCTTC
12
803
|
|
1131753
2014
2033
7402
7421
CACTTTATTGAGTTCCTGCG
15
163
|
|
1131781
N/A
N/A
115
134
GGACAATCCTGGTTCCCACA
71
804
|
|
1131809
N/A
N/A
199
218
AGGCCTCCTAGTCACCTGGA
58
805
|
|
1131837
N/A
N/A
302
321
GGCCCATCTCCCCCAGAAAT
50
806
|
|
1131865
N/A
N/A
392
411
CTAGTCTGCAGCTTCCTCTC
48
807
|
|
1131893
N/A
N/A
519
538
CACTTACCGACTGTGTGCTC
24
808
|
|
1131921
N/A
N/A
627
646
GGGCATAAGACCTAGCACCA
46
809
|
|
1131949
N/A
N/A
730
749
ACATATCTCAAGCACCTGCT
63
810
|
|
1131977
N/A
N/A
880
899
GCTCTTGATCCTTCCTTGCC
54
811
|
|
1132005
N/A
N/A
997
1016
AACCTGGTTTCACAGAAGGG
54
812
|
|
1132033
N/A
N/A
1085
1104
TTGTAAAGCATTTCCATGCA
37
813
|
|
1132061
N/A
N/A
1512
1531
ACTCTAGGTAAATTTTTTTT
86
814
|
|
1132089
N/A
N/A
1612
1631
AATGTGCTGGGCCTCAACGC
49
815
|
|
1132117
N/A
N/A
1908
1927
AAGTTGTGTGCCTGTAATCC
50
816
|
|
1132145
N/A
N/A
2144
2163
CAAGGCAAAGTGCACTTGCC
39
817
|
|
1132173
N/A
N/A
2274
2293
GCGAATAATGAGAATTGCCT
38
818
|
|
1132201
N/A
N/A
2388
2407
TAAAACCAGATTATATGCTG
67
819
|
|
1132229
N/A
N/A
2522
2541
CACAAGAAGGCTACCTTAGG
55
820
|
|
1132257
N/A
N/A
2649
2668
CCTATTGAATGAGTGTCCTT
48
821
|
|
1132285
N/A
N/A
2760
2779
CTTGAAATGTTTTCATGGCC
59
822
|
|
1132313
N/A
N/A
2854
2873
AGTAATAATAGTACAGTTGA
59
823
|
|
1132341
N/A
N/A
2965
2984
GGCCAGGAAGAGCACTTGCC
59
824
|
|
1132369
N/A
N/A
3373
3392
CTGCTCTAAAAGTTGGGTTC
40
825
|
|
1132397
N/A
N/A
3460
3479
TACTCAACTGCTCAGGCACT
34
826
|
|
1132425
N/A
N/A
3658
3677
AGAAGGACAGGCAGGGTACA
67
827
|
|
1132453
N/A
N/A
3824
3843
CCTTTCACTTTCTTGGGCTC
16
828
|
|
1132481
N/A
N/A
3925
3944
GGCTGGGTCCAGAATCCCAG
40
829
|
|
1132509
N/A
N/A
4036
4055
ACCTTGCTACTCCAAGTTTC
36
830
|
|
1132537
N/A
N/A
4236
4255
CCTTTCTGGCAGTGGTTTCC
20
831
|
|
1132565
N/A
N/A
4337
4356
CTAGGACCATGCCAAGTCTC
89
832
|
|
1132593
N/A
N/A
4569
4588
CCCGGCCTCCTGCCAGCCTG
40
833
|
|
1132621
N/A
N/A
4769
4788
TTGCCCCAGACCCTCACTCA
47
834
|
|
1132649
N/A
N/A
4902
4921
CCTTGGTGTCTGAGGAGAAA
59
835
|
|
1132677
N/A
N/A
5418
5437
GAATCTAGCTCGCCCGGCGC
58
836
|
|
1132705
N/A
N/A
5844
5863
TTCCTCCCCGGGAGCTCCGG
65
837
|
|
1132733
N/A
N/A
6306
6325
ACCCTCGAACTGGTGGCCCC
59
838
|
|
1132761
N/A
N/A
6397
6416
ACCCGGAACGATACCAAAGT
61
839
|
|
1132789
N/A
N/A
6518
6537
CTCCCTGTATCCCCAGGAGA
77
840
|
|
1132817
N/A
N/A
6643
6662
CCCCTCCCCCACCACCCATC
46
841
|
|
1132845
N/A
N/A
6733
6752
GGAGGAGGTGCCATTAATTC
61
842
|
|
1132873
N/A
N/A
6874
6893
CATATTCCTCCGCCCCTGCG
90
843
|
|
TABLE 15
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed
|
PO/PS internucleoside linkages (Huh7, electroporation,
|
5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
FXII
SEQ ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130745
5
24
5
24
CTATCCAGGAGTCCAGATCA
58
844
|
|
1130773
42
61
42
61
CAGAGCCCTCATGGCATCCG
33
845
|
|
1130801
90
109
N/A
N/A
AATCGAAAGTGTTGACTCCA
33
846
|
|
1130829
146
165
N/A
N/A
ACGACTGTGTGCTCTTCAGC
20
176
|
|
1130857
200
219
3551
3570
AGCTGCCGGTGGTACTGGAA
43
847
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
18
183
|
|
1130886
235
254
3586
3605
GGCCTGGCCGGCCCTTGTGG
49
848
|
|
1130914
305
324
3812
3831
TTGGGCTCCAAACAGTATCC
20
849
|
|
1130942
361
380
4169
4188
CACAGGTCCCTCCTTTCTGG
62
850
|
|
1130970
417
436
4225
4244
GTGGTTTCCAGTGAGGTGTT
19
851
|
|
1130998
459
478
4404
4423
AAACCGGAGAAGCTGAGGCT
21
852
|
|
1131026
501
520
4446
4465
AGCTGCTTGCTCAGTTCTAT
41
853
|
|
1131054
542
561
4487
4506
TGGCAGTGGGCATCAGGACC
60
854
|
|
1131082
577
596
N/A
N/A
ACGGGTTGGTGCGGCAGGCC
31
855
|
|
1131110
617
636
4701
4720
CGGTGGCCCTCCACCTCTAG
44
856
|
|
1131138
679
698
N/A
N/A
AGCTTGCCTTGGTGTCCACG
38
857
|
|
1131166
719
738
4948
4967
CTGGCCAGGCCGCGGTAGCT
92
858
|
|
1131194
774
793
5003
5022
CCGGTAGGTGGCCTCCGAGG
44
859
|
|
1131222
868
887
5182
5201
GCACGAAGCACCACGGGCGG
30
860
|
|
1131250
912
931
5226
5245
CTGTGCCAGGTCGCAGTACT
38
861
|
|
1131278
1003
1022
5317
5336
GCTTCGGCGGTGCCGGCTGC
33
862
|
|
1131306
1080
1099
5499
5518
GGAAGGCGGCTGCTCCCGCT
36
863
|
|
1131334
1127
1146
5546
5565
CTCTTGCGGAGCCGCTGCCC
46
864
|
|
1131362
1164
1183
5583
5602
CACCAGCCCGCCAACGACGC
24
865
|
|
1131390
1234
1253
5653
5672
TGAGGCTGCCGGCGCAGAAA
40
866
|
|
1131418
1319
1338
5979
5998
TCCTGGCCGAGCACCACCGT
34
867
|
|
1131446
1410
1429
6070
6089
GTGCTGGTAGCTGACGGGCG
26
868
|
|
1131474
1470
1489
6213
6232
AGGCGACAGGAGCGCGCAGC
21
869
|
|
1131502
1556
1575
6299
6318
AACTGGTGGCCCCAGCCGGC
47
870
|
|
1131530
1588
1607
6886
6905
GCAGGAAGCTGGCATATTCC
58
871
|
|
1131558
1681
1700
6979
6998
ACCCTGCGCAGAGCATGCCG
20†
872
|
|
1131586
1755
1774
7143
7162
CTCTGCAGCTTGGTCCTCAC
14†
873
|
|
1131614
1801
1820
7189
7208
CACAGCCCGATCCCCAGCTG
38
874
|
|
1131642
1848
1867
7236
7255
CAGGTAGTAGGCCACATCGG
35
875
|
|
1131670
1881
1900
7269
7288
CAATCAGGAAACGGTGTGCT
47
876
|
|
1131698
1924
1943
7312
7331
TCTCACTGCGGAATCACCAA
6
877
|
|
1131726
1961
1980
7349
7368
ATGGGACACAATCTTGCCTT
34
878
|
|
1131754
2015
2034
7403
7422
GCACTTTATTGAGTTCCTGC
4
164
|
|
1131782
N/A
N/A
118
137
CTGGGACAATCCTGGTTCCC
56
879
|
|
1131810
N/A
N/A
202
221
CATAGGCCTCCTAGTCACCT
94
880
|
|
1131838
N/A
N/A
305
324
TAAGGCCCATCTCCCCCAGA
41
881
|
|
1131866
N/A
N/A
395
414
TTGCTAGTCTGCAGCTTCCT
60
882
|
|
1131894
N/A
N/A
523
542
AGGCCACTTACCGACTGTGT
109
883
|
|
1131922
N/A
N/A
630
649
CATGGGCATAAGACCTAGCA
65
884
|
|
1131950
N/A
N/A
733
752
AGCACATATCTCAAGCACCT
85
885
|
|
1131978
N/A
N/A
886
905
AGACAAGCTCTTGATCCTTC
79
886
|
|
1132006
N/A
N/A
1000
1019
ATCAACCTGGTTTCACAGAA
48
887
|
|
1132034
N/A
N/A
1088
1107
TATTTGTAAAGCATTTCCAT
67
888
|
|
1132062
N/A
N/A
1515
1534
CACACTCTAGGTAAATTTTT
59
889
|
|
1132090
N/A
N/A
1615
1634
TGGAATGTGCTGGGCCTCAA
59
890
|
|
1132118
N/A
N/A
1911
1930
ATGAAGTTGTGTGCCTGTAA
61
891
|
|
1132146
N/A
N/A
2147
2166
GTTCAAGGCAAAGTGCACTT
56
892
|
|
1132174
N/A
N/A
2277
2296
ATCGCGAATAATGAGAATTG
68
893
|
|
1132202
N/A
N/A
2391
2410
ACATAAAACCAGATTATATG
52
894
|
|
1132230
N/A
N/A
2525
2544
AAGCACAAGAAGGCTACCTT
33
895
|
|
1132258
N/A
N/A
2652
2671
TCTCCTATTGAATGAGTGTC
36
896
|
|
1132286
N/A
N/A
2765
2784
CAATACTTGAAATGTTTTCA
47
897
|
|
1132314
N/A
N/A
2857
2876
GGCAGTAATAATAGTACAGT
38
898
|
|
1132342
N/A
N/A
2968
2987
CAAGGCCAGGAAGAGCACTT
76
899
|
|
1132370
N/A
N/A
3376
3395
TTTCTGCTCTAAAAGTTGGG
35
900
|
|
1132398
N/A
N/A
3463
3482
CTGTACTCAACTGCTCAGGC
60
901
|
|
1132426
N/A
N/A
3661
3680
GACAGAAGGACAGGCAGGGT
52
902
|
|
1132454
N/A
N/A
3827
3846
GCACCTTTCACTTTCTTGGG
24
903
|
|
1132482
N/A
N/A
3945
3964
TATTGTGGAGGGAGAGAAGG
83
904
|
|
1132510
N/A
N/A
4039
4058
GTGACCTTGCTACTCCAAGT
74
905
|
|
1132538
N/A
N/A
4239
4258
TCACCTTTCTGGCAGTGGTT
44
906
|
|
1132566
N/A
N/A
4340
4359
AGTCTAGGACCATGCCAAGT
34
907
|
|
1132594
N/A
N/A
4572
4591
CCACCCGGCCTCCTGCCAGC
75
908
|
|
1132622
N/A
N/A
4772
4791
TGCTTGCCCCAGACCCTCAC
54
909
|
|
1132650
N/A
N/A
4905
4924
TTGCCTTGGTGTCTGAGGAG
44
910
|
|
1132678
N/A
N/A
5421
5440
CCGGAATCTAGCTCGCCCGG
50
911
|
|
1132706
N/A
N/A
5847
5866
AGCTTCCTCCCCGGGAGCTC
64
912
|
|
1132734
N/A
N/A
6309
6328
CCTACCCTCGAACTGGTGGC
61
913
|
|
1132762
N/A
N/A
6400
6419
GGCACCCGGAACGATACCAA
63
914
|
|
1132790
N/A
N/A
6521
6540
ATTCTCCCTGTATCCCCAGG
58
915
|
|
1132818
N/A
N/A
6646
6665
AAACCCCTCCCCCACCACCC
41
916
|
|
1132846
N/A
N/A
6752
6771
CTCGCAGCAAGCCCGAAGGG
38
917
|
|
1132874
N/A
N/A
6877
6896
TGGCATATTCCTCCGCCCCT
49
918
|
|
TABLE 16
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed
|
PO/PS internucleoside linkages (Huh7, electroporation,
|
5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
FXII
SEQ ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130746
7
26
7
26
GCCTATCCAGGAGTCCAGAT
29
919
|
|
1130774
46
65
46
65
GCAGCAGAGCCCTCATGGCA
22
920
|
|
1130802
91
110
N/A
N/A
GAATCGAAAGTGTTGACTCC
40
921
|
|
1130830
147
166
N/A
N/A
AACGACTGTGTGCTCTTCAG
9
922
|
|
1130858
201
220
3552
3571
CAGCTGCCGGTGGTACTGGA
22
923
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
22
183
|
|
1130887
236
255
3587
3606
GGGCCTGGCCGGCCCTTGTG
41
924
|
|
1130915
306
325
3813
3832
CTTGGGCTCCAAACAGTATC
26
925
|
|
1130943
362
381
4170
4189
ACACAGGTCCCTCCTTTCTG
32
926
|
|
1130971
419
438
4227
4246
CAGTGGTTTCCAGTGAGGTG
38
927
|
|
1130999
460
479
4405
4424
AAAACCGGAGAAGCTGAGGC
26
928
|
|
1131027
502
521
4447
4466
CAGCTGCTTGCTCAGTTCTA
35
929
|
|
1131055
543
562
4488
4507
CTGGCAGTGGGCATCAGGAC
33
930
|
|
1131083
579
598
4663
4682
GCACGGGTTGGTGCGGCAGG
29
931
|
|
1131111
618
637
4702
4721
GCGGTGGCCCTCCACCTCTA
54
932
|
|
1131139
680
699
N/A
N/A
CAGCTTGCCTTGGTGTCCAC
30
933
|
|
1131167
720
739
4949
4968
CCTGGCCAGGCCGCGGTAGC
80
934
|
|
1131195
775
794
5004
5023
TCCGGTAGGTGGCCTCCGAG
33
935
|
|
1131223
869
888
5183
5202
AGCACGAAGCACCACGGGCG
17
936
|
|
1131251
913
932
5227
5246
ACTGTGCCAGGTCGCAGTAC
26
937
|
|
1131279
1004
1023
5318
5337
GGCTTCGGCGGTGCCGGCTG
24
938
|
|
1131307
1081
1100
5500
5519
GGGAAGGCGGCTGCTCCCGC
40
939
|
|
1131335
1128
1147
5547
5566
ACTCTTGCGGAGCCGCTGCC
43
98
|
|
1131363
1165
1184
5584
5603
CCACCAGCCCGCCAACGACG
59
940
|
|
1131391
1235
1254
5654
5673
ATGAGGCTGCCGGCGCAGAA
49
941
|
|
1131419
1320
1339
5980
5999
TTCCTGGCCGAGCACCACCG
24
942
|
|
1131447
1411
1430
6071
6090
CGTGCTGGTAGCTGACGGGC
25
943
|
|
1131475
1471
1490
6214
6233
AAGGCGACAGGAGCGCGCAG
28
944
|
|
1131503
1557
1576
6300
6319
GAACTGGTGGCCCCAGCCGG
65
945
|
|
1131531
1592
1611
6890
6909
TCCTGCAGGAAGCTGGCATA
88
946
|
|
1131559
1683
1702
6981
7000
GAACCCTGCGCAGAGCATGC
56†
947
|
|
1131587
1757
1776
7145
7164
CGCTCTGCAGCTTGGTCCTC
19†
948
|
|
1131615
1802
1821
7190
7209
CCACAGCCCGATCCCCAGCT
40
949
|
|
1131643
1849
1868
7237
7256
CCAGGTAGTAGGCCACATCG
44
88
|
|
1131671
1882
1901
7270
7289
GCAATCAGGAAACGGTGTGC
12
950
|
|
1131699
1926
1945
7314
7333
TCTCTCACTGCGGAATCACC
7
951
|
|
1131727
1962
1981
7350
7369
AATGGGACACAATCTTGCCT
25
952
|
|
1131755
2016
2035
7404
7423
AGCACTTTATTGAGTTCCTG
3
953
|
|
1131783
N/A
N/A
121
140
ATCCTGGGACAATCCTGGTT
41
954
|
|
1131811
N/A
N/A
205
224
CCACATAGGCCTCCTAGTCA
48
955
|
|
1131839
N/A
N/A
308
327
TCTTAAGGCCCATCTCCCCC
46
956
|
|
1131867
N/A
N/A
398
417
CTGTTGCTAGTCTGCAGCTT
85
957
|
|
1131895
N/A
N/A
526
545
GCCAGGCCACTTACCGACTG
40
958
|
|
1131923
N/A
N/A
633
652
GCCCATGGGCATAAGACCTA
37
959
|
|
1131951
N/A
N/A
736
755
AGCAGCACATATCTCAAGCA
68
960
|
|
1131979
N/A
N/A
889
908
CAGAGACAAGCTCTTGATCC
44
961
|
|
1132007
N/A
N/A
1003
1022
GTTATCAACCTGGTTTCACA
27
962
|
|
1132035
N/A
N/A
1097
1116
AAAAGCTAATATTTGTAAAG
42
963
|
|
1132063
N/A
N/A
1518
1537
TGCCACACTCTAGGTAAATT
43
964
|
|
1132091
N/A
N/A
1618
1637
GGTTGGAATGTGCTGGGCCT
48
965
|
|
1132119
N/A
N/A
1914
1933
AGCATGAAGTTGTGTGCCTG
79
966
|
|
1132147
N/A
N/A
2150
2169
TTTGTTCAAGGCAAAGTGCA
34
967
|
|
1132175
N/A
N/A
2280
2299
ACCATCGCGAATAATGAGAA
32
968
|
|
1132203
N/A
N/A
2394
2413
CATACATAAAACCAGATTAT
67
969
|
|
1132231
N/A
N/A
2533
2552
TAGTTCCTAAGCACAAGAAG
98
970
|
|
1132259
N/A
N/A
2655
2674
AGCTCTCCTATTGAATGAGT
36
971
|
|
1132287
N/A
N/A
2768
2787
AGTCAATACTTGAAATGTTT
84
972
|
|
1132315
N/A
N/A
2860
2879
AACGGCAGTAATAATAGTAC
55
973
|
|
1132343
N/A
N/A
2971
2990
TCCCAAGGCCAGGAAGAGCA
32
974
|
|
1132371
N/A
N/A
3379
3398
CACTTTCTGCTCTAAAAGTT
28
975
|
|
1132399
N/A
N/A
3466
3485
CCCCTGTACTCAACTGCTCA
42
976
|
|
1132427
N/A
N/A
3664
3683
TTGGACAGAAGGACAGGCAG
58
977
|
|
1132455
N/A
N/A
3830
3849
GTAGCACCTTTCACTTTCTT
16
978
|
|
1132483
N/A
N/A
3954
3973
TCCAAAGGGTATTGTGGAGG
38
979
|
|
1132511
N/A
N/A
4042
4061
CTTGTGACCTTGCTACTCCA
28
980
|
|
1132539
N/A
N/A
4242
4261
TCCTCACCTTTCTGGCAGTG
27
981
|
|
1132567
N/A
N/A
4343
4362
GAGAGTCTAGGACCATGCCA
80
982
|
|
1132595
N/A
N/A
4575
4594
ACACCACCCGGCCTCCTGCC
58
983
|
|
1132623
N/A
N/A
4775
4794
TTCTGCTTGCCCCAGACCCT
70
984
|
|
1132651
N/A
N/A
4908
4927
AGCTTGCCTTGGTGTCTGAG
19
985
|
|
1132679
N/A
N/A
5424
5443
TGGCCGGAATCTAGCTCGCC
61
986
|
|
1132707
N/A
N/A
5850
5869
TCCAGCTTCCTCCCCGGGAG
51
987
|
|
1132735
N/A
N/A
6312
6331
GTGCCTACCCTCGAACTGGT
45
988
|
|
1132763
N/A
N/A
6403
6422
GTAGGCACCCGGAACGATAC
27
989
|
|
1132791
N/A
N/A
6524
6543
TCGATTCTCCCTGTATCCCC
69
990
|
|
1132819
N/A
N/A
6649
6668
CAGAAACCCCTCCCCCACCA
50
991
|
|
1132847
N/A
N/A
6755
6774
TCTCTCGCAGCAAGCCCGAA
48
992
|
|
1132875
N/A
N/A
7010
7029
ACCTGGCACGCATCGGTGCC
29†
993
|
|
TABLE 17
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed
|
PO/PS internucleoside linkages (Huh7, electroporation,
|
5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
FXII
SEQ ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130747
10
29
10
29
GCTGCCTATCCAGGAGTCCA
45
994
|
|
1130775
47
66
47
66
AGCAGCAGAGCCCTCATGGC
43
995
|
|
1130803
93
112
N/A
N/A
TGGAATCGAAAGTGTTGACT
25
996
|
|
1130831
148
167
N/A
N/A
GAACGACTGTGTGCTCTTCA
18
997
|
|
1130859
203
222
3554
3573
TACAGCTGCCGGTGGTACTG
28
998
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
8
183
|
|
1130888
238
257
3589
3608
GAGGGCCTGGCCGGCCCTTG
77
999
|
|
1130916
307
326
3814
3833
TCTTGGGCTCCAAACAGTAT
33
1000
|
|
1130944
363
382
4171
4190
CACACAGGTCCCTCCTTTCT
53
1001
|
|
1130972
420
439
4228
4247
GCAGTGGTTTCCAGTGAGGT
19
1002
|
|
1131000
461
480
4406
4425
AAAAACCGGAGAAGCTGAGG
25
1003
|
|
1131028
505
524
4450
4469
CCACAGCTGCTTGCTCAGTT
23
1004
|
|
1131056
545
564
4490
4509
CGCTGGCAGTGGGCATCAGG
24
1005
|
|
1131084
580
599
4664
4683
GGCACGGGTTGGTGCGGCAG
19
1006
|
|
1131112
619
638
4703
4722
GGCGGTGGCCCTCCACCTCT
36
1007
|
|
1131140
681
700
N/A
N/A
GCAGCTTGCCTTGGTGTCCA
21
1008
|
|
1131168
721
740
4950
4969
TCCTGGCCAGGCCGCGGTAG
37
1009
|
|
1131196
777
796
5006
5025
GTTCCGGTAGGTGGCCTCCG
46
1010
|
|
1131224
870
889
5184
5203
CAGCACGAAGCACCACGGGC
35
1011
|
|
1131252
914
933
5228
5247
CACTGTGCCAGGTCGCAGTA
60
1012
|
|
1131280
1005
1024
5319
5338
AGGCTTCGGCGGTGCCGGCT
31
1013
|
|
1131308
1085
1104
5504
5523
GTCAGGGAAGGCGGCTGCTC
25
1014
|
|
1131336
1129
1148
5548
5567
GACTCTTGCGGAGCCGCTGC
40
1015
|
|
1131364
1167
1186
5586
5605
CGCCACCAGCCCGCCAACGA
43
1016
|
|
1131392
1237
1256
5656
5675
CGATGAGGCTGCCGGCGCAG
30
1017
|
|
1131420
1322
1341
5982
6001
CGTTCCTGGCCGAGCACCAC
34
1018
|
|
1131448
1430
1449
N/A
N/A
AGGCGCAACAGAGCCAGGTC
38
1019
|
|
1131476
1473
1492
6216
6235
GTAAGGCGACAGGAGCGCGC
37
1020
|
|
1131504
1558
1577
6301
6320
CGAACTGGTGGCCCCAGCCG
76
1021
|
|
1131532
1594
1613
6892
6911
CCTCCTGCAGGAAGCTGGCA
112
1022
|
|
1131560
1687
1706
6985
7004
CGAGGAACCCTGCGCAGAGC
6†
1023
|
|
1131588
1758
1777
7146
7165
GCGCTCTGCAGCTTGGTCCT
16†
1024
|
|
1131616
1803
1822
7191
7210
ACCACAGCCCGATCCCCAGC
39
1025
|
|
1131644
1850
1869
7238
7257
GCCAGGTAGTAGGCCACATC
34
1026
|
|
1131672
1883
1902
7271
7290
AGCAATCAGGAAACGGTGTG
24
1027
|
|
1131700
1927
1946
7315
7334
CTCTCTCACTGCGGAATCAC
7
1028
|
|
1131728
1963
1982
7351
7370
GAATGGGACACAATCTTGCC
21
1029
|
|
1131756
2017
2036
7405
7424
AAGCACTTTATTGAGTTCCT
5
1030
|
|
1131784
N/A
N/A
124
143
ACAATCCTGGGACAATCCTG
85
1031
|
|
1131812
N/A
N/A
208
227
TTTCCACATAGGCCTCCTAG
42
1032
|
|
1131840
N/A
N/A
311
330
TCTTCTTAAGGCCCATCTCC
79
1033
|
|
1131868
N/A
N/A
401
420
GATCTGTTGCTAGTCTGCAG
82
1034
|
|
1131896
N/A
N/A
529
548
GGAGCCAGGCCACTTACCGA
45
1035
|
|
1131924
N/A
N/A
636
655
AGTGCCCATGGGCATAAGAC
52
1036
|
|
1131952
N/A
N/A
739
758
GCCAGCAGCACATATCTCAA
55
1037
|
|
1131980
N/A
N/A
892
911
TGCCAGAGACAAGCTCTTGA
73
1038
|
|
1132008
N/A
N/A
1006
1025
ACTGTTATCAACCTGGTTTC
60
1039
|
|
1132036
N/A
N/A
1105
1124
TTCATAATAAAAGCTAATAT
68
1040
|
|
1132064
N/A
N/A
1521
1540
ATGTGCCACACTCTAGGTAA
51
1041
|
|
1132092
N/A
N/A
1622
1641
CCAGGGTTGGAATGTGCTGG
56
1042
|
|
1132120
N/A
N/A
1917
1936
CTGAGCATGAAGTTGTGTGC
44
1043
|
|
1132148
N/A
N/A
2166
2185
AAGCTTTTGCCACTATTTTG
36
1044
|
|
1132176
N/A
N/A
2283
2302
ATAACCATCGCGAATAATGA
50
1045
|
|
1132204
N/A
N/A
2397
2416
AATCATACATAAAACCAGAT
75
1046
|
|
1132232
N/A
N/A
2536
2555
GTGTAGTTCCTAAGCACAAG
37
1047
|
|
1132260
N/A
N/A
2658
2677
TTCAGCTCTCCTATTGAATG
37
1048
|
|
1132288
N/A
N/A
2771
2790
CCAAGTCAATACTTGAAATG
42
1049
|
|
1132316
N/A
N/A
2863
2882
TAAAACGGCAGTAATAATAG
62
1050
|
|
1132344
N/A
N/A
2974
2993
CAGTCCCAAGGCCAGGAAGA
38
1051
|
|
1132372
N/A
N/A
3382
3401
AAACACTTTCTGCTCTAAAA
67
1052
|
|
1132400
N/A
N/A
3469
3488
CTTCCCCTGTACTCAACTGC
43
1053
|
|
1132428
N/A
N/A
3667
3686
TCCTTGGACAGAAGGACAGG
75
1054
|
|
1132456
N/A
N/A
3833
3852
TGTGTAGCACCTTTCACTTT
41
1055
|
|
1132484
N/A
N/A
3957
3976
ACTTCCAAAGGGTATTGTGG
56
1056
|
|
1132512
N/A
N/A
4045
4064
TGCCTTGTGACCTTGCTACT
34
1057
|
|
1132540
N/A
N/A
4245
4264
ATCTCCTCACCTTTCTGGCA
82
1058
|
|
1132568
N/A
N/A
4346
4365
CAGGAGAGTCTAGGACCATG
78
1059
|
|
1132596
N/A
N/A
4578
4597
GGCACACCACCCGGCCTCCT
48
1060
|
|
1132624
N/A
N/A
4778
4797
GCCTTCTGCTTGCCCCAGAC
48
1061
|
|
1132652
N/A
N/A
4911
4930
AGCAGCTTGCCTTGGTGTCT
25
1062
|
|
1132680
N/A
N/A
5427
5446
GGCTGGCCGGAATCTAGCTC
69
1063
|
|
1132708
N/A
N/A
5853
5872
TGTTCCAGCTTCCTCCCCGG
37
1064
|
|
1132736
N/A
N/A
6315
6334
GTTGTGCCTACCCTCGAACT
85
1065
|
|
1132764
N/A
N/A
6406
6425
TCTGTAGGCACCCGGAACGA
58
1066
|
|
1132792
N/A
N/A
6527
6546
AGTTCGATTCTCCCTGTATC
54
1067
|
|
1132820
N/A
N/A
6652
6671
AAACAGAAACCCCTCCCCCA
73
1068
|
|
1132848
N/A
N/A
6758
6777
TCCTCTCTCGCAGCAAGCCC
57
1069
|
|
1132876
N/A
N/A
7013
7032
CTCACCTGGCACGCATCGGT
64†
1070
|
|
TABLE 18
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed
|
PO/PS internucleoside linkages (Huh7, electroporation,
|
5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
FXII
SEQ ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130749
12
31
12
31
CAGCTGCCTATCCAGGAGTC
57
1071
|
|
1130777
49
68
49
68
GGAGCAGCAGAGCCCTCATG
40
1072
|
|
1130805
96
115
N/A
N/A
AGGTGGAATCGAAAGTGTTG
34
1073
|
|
1130833
151
170
N/A
N/A
TGAGAACGACTGTGTGCTCT
27
1074
|
|
1130861
205
224
3556
3575
GGTACAGCTGCCGGTGGTAC
31
1075
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
10
183
|
|
1130890
240
259
3591
3610
CTGAGGGCCTGGCCGGCCCT
40
1076
|
|
1130918
310
329
3817
3836
CTTTCTTGGGCTCCAAACAG
36
1077
|
|
1130946
366
385
4174
4193
GTTCACACAGGTCCCTCCTT
48
1078
|
|
1130974
423
442
4231
4250
CTGGCAGTGGTTTCCAGTGA
28
1079
|
|
1131002
464
483
4409
4428
TGGAAAAACCGGAGAAGCTG
22
1080
|
|
1131030
508
527
4453
4472
TGGCCACAGCTGCTTGCTCA
48
1081
|
|
1131058
547
566
4492
4511
GCCGCTGGCAGTGGGCATCA
42
1082
|
|
1131086
583
602
4667
4686
GGAGGCACGGGTTGGTGCGG
27
1083
|
|
1131114
622
641
4706
4725
ACAGGCGGTGGCCCTCCACC
58
1084
|
|
1131142
685
704
4914
4933
CATAGCAGCTTGCCTTGGTG
22
1085
|
|
1131170
724
743
4953
4972
TGGTCCTGGCCAGGCCGCGG
77
1086
|
|
1131198
796
815
5025
5044
GCGCTTGCTCGGCAGTCACG
18
1087
|
|
1131226
873
892
5187
5206
GTTCAGCACGAAGCACCACG
21
1088
|
|
1131254
916
935
5230
5249
GGCACTGTGCCAGGTCGCAG
39
1089
|
|
1131282
1007
1026
5321
5340
TGAGGCTTCGGCGGTGCCGG
43
1090
|
|
1131310
1089
1108
5508
5527
CCTGGTCAGGGAAGGCGGCT
27
1091
|
|
1131338
1131
1150
5550
5569
CAGACTCTTGCGGAGCCGCT
22
1092
|
|
1131366
1203
1222
5622
5641
GTACAGCGCGGCGATGTAGG
41
1093
|
|
1131394
1239
1258
5658
5677
GGCGATGAGGCTGCCGGCGC
31
114
|
|
1131422
1341
1360
6001
6020
CTCACAGCTGTGGTTACGGC
23
1094
|
|
1131450
1433
1452
N/A
N/A
TGAAGGCGCAACAGAGCCAG
46
1095
|
|
1131478
1492
1511
6235
6254
GCAGGCACACCGGCTGAACG
36
72
|
|
1131506
1560
1579
6303
6322
CTCGAACTGGTGGCCCCAGC
53
1096
|
|
1131534
1597
1616
6895
6914
GCGCCTCCTGCAGGAAGCTG
92
1097
|
|
1131562
1690
1709
6988
7007
CCTCGAGGAACCCTGCGCAG
27†
1098
|
|
1131590
1776
1795
7164
7183
GCCTTGCAGGGTGAGCCGGC
43
1099
|
|
1131618
1805
1824
7193
7212
TCACCACAGCCCGATCCCCA
40
1100
|
|
1131646
1852
1871
7240
7259
AGGCCAGGTAGTAGGCCACA
63
1101
|
|
1131674
1888
1907
7276
7295
CCCTGAGCAATCAGGAAACG
35
1102
|
|
1131702
1930
1949
7318
7337
CCACTCTCTCACTGCGGAAT
9
1103
|
|
1131730
1984
2003
7372
7391
GCGGAGCTGGCCGCACTGGG
18
1104
|
|
1131758
2019
2038
7407
7426
CAAAGCACTTTATTGAGTTC
9
1105
|
|
1131786
N/A
N/A
130
149
CCCAGAACAATCCTGGGACA
70
1106
|
|
1131814
N/A
N/A
214
233
CTCACCTTTCCACATAGGCC
82
1107
|
|
1131842
N/A
N/A
317
336
CCGTTGTCTTCTTAAGGCCC
82
1108
|
|
1131870
N/A
N/A
407
426
GCCACCGATCTGTTGCTAGT
67
1109
|
|
1131898
N/A
N/A
538
557
CCGGGAGGAGGAGCCAGGCC
67
1110
|
|
1131926
N/A
N/A
642
661
CACTCTAGTGCCCATGGGCA
83
1111
|
|
1131954
N/A
N/A
760
779
CAGGAGCCCAGGTGTGATGG
47
1112
|
|
1131982
N/A
N/A
898
917
TTCAGATGCCAGAGACAAGC
54
1113
|
|
1132010
N/A
N/A
1012
1031
AGGTTTACTGTTATCAACCT
48
1114
|
|
1132038
N/A
N/A
1111
1130
GGTAGTTTCATAATAAAAGC
36
1115
|
|
1132066
N/A
N/A
1527
1546
CCTGCTATGTGCCACACTCT
59
1116
|
|
1132094
N/A
N/A
1637
1656
GGAGGCTGAGGGAGTCCAGG
90
1117
|
|
1132122
N/A
N/A
1923
1942
AATTAGCTGAGCATGAAGTT
54
1118
|
|
1132150
N/A
N/A
2172
2191
CTCAATAAGCTTTTGCCACT
57
1119
|
|
1132178
N/A
N/A
2289
2308
TAGAACATAACCATCGCGAA
33
1120
|
|
1132206
N/A
N/A
2411
2430
ATGTCTTTAAACAGAATCAT
32
1121
|
|
1132234
N/A
N/A
2542
2561
AGAGCTGTGTAGTTCCTAAG
65
1122
|
|
1132262
N/A
N/A
2664
2683
GCTTGTTTCAGCTCTCCTAT
78
1123
|
|
1132290
N/A
N/A
2777
2796
TAACTCCCAAGTCAATACTT
76
1124
|
|
1132318
N/A
N/A
2890
2909
TGATCTGAGCCTTGGTTTCC
69
1125
|
|
1132346
N/A
N/A
2980
2999
AGTCTTCAGTCCCAAGGCCA
71
1126
|
|
1132374
N/A
N/A
3388
3407
CATTGAAAACACTTTCTGCT
70
1127
|
|
1132402
N/A
N/A
3475
3494
TGAGGACTTCCCCTGTACTC
40
1128
|
|
1132430
N/A
N/A
3673
3692
CAGAGTTCCTTGGACAGAAG
38
1129
|
|
1132458
N/A
N/A
3839
3858
AGGCTGTGTGTAGCACCTTT
54
1130
|
|
1132486
N/A
N/A
3963
3982
CTCTGGACTTCCAAAGGGTA
37
1131
|
|
1132514
N/A
N/A
4051
4070
CCTACTTGCCTTGTGACCTT
37
1132
|
|
1132542
N/A
N/A
4251
4270
CTCCACATCTCCTCACCTTT
39
1133
|
|
1132570
N/A
N/A
4352
4371
TGGTCTCAGGAGAGTCTAGG
59
1134
|
|
1132598
N/A
N/A
4584
4603
CTTCCTGGCACACCACCCGG
60
1135
|
|
1132626
N/A
N/A
4784
4803
GGGCTGGCCTTCTGCTTGCC
36
1136
|
|
1132654
N/A
N/A
5070
5089
ACGCGGCGCACCGGCAGAAG
61
1137
|
|
1132682
N/A
N/A
5436
5455
CCCGCGGCCGGCTGGCCGGA
118
1138
|
|
1132710
N/A
N/A
5859
5878
ATCCCGTGTTCCAGCTTCCT
67
1139
|
|
1132738
N/A
N/A
6321
6340
CTAGCAGTTGTGCCTACCCT
49
1140
|
|
1132766
N/A
N/A
6412
6431
ACCCATTCTGTAGGCACCCG
78
1141
|
|
1132794
N/A
N/A
6533
6552
AGAGCAAGTTCGATTCTCCC
88
1142
|
|
1132822
N/A
N/A
6658
6677
GAGCGGAAACAGAAACCCCT
52
1143
|
|
1132850
N/A
N/A
6764
6783
TGCCCTTCCTCTCTCGCAGC
95
1144
|
|
1132878
N/A
N/A
7019
7038
TAAGAGCTCACCTGGCACGC
85†
1145
|
|
TABLE 19
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed
|
PO/PS internucleoside linkages (Huh7, electroporation,
|
5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
FXII
SEQ ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130748
11
30
11
30
AGCTGCCTATCCAGGAGTCC
62
1146
|
|
1130776
48
67
48
67
GAGCAGCAGAGCCCTCATGG
67
1147
|
|
1130804
94
113
N/A
N/A
GTGGAATCGAAAGTGTTGAC
58
1148
|
|
1130832
150
169
N/A
N/A
GAGAACGACTGTGTGCTCTT
12
1149
|
|
1130860
204
223
3555
3574
GTACAGCTGCCGGTGGTACT
35
1150
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
16
183
|
|
1130889
239
258
3590
3609
TGAGGGCCTGGCCGGCCCTT
54
1151
|
|
1130917
308
327
3815
3834
TTCTTGGGCTCCAAACAGTA
25
1152
|
|
1130945
365
384
4173
4192
TTCACACAGGTCCCTCCTTT
37
1153
|
|
1130973
421
440
4229
4248
GGCAGTGGTTTCCAGTGAGG
14
1154
|
|
1131001
463
482
4408
4427
GGAAAAACCGGAGAAGCTGA
26
1155
|
|
1131029
506
525
4451
4470
GCCACAGCTGCTTGCTCAGT
35
1156
|
|
1131057
546
565
4491
4510
CCGCTGGCAGTGGGCATCAG
32
1157
|
|
1131085
582
601
4666
4685
GAGGCACGGGTTGGTGCGGC
32
1158
|
|
1131113
621
640
4705
4724
CAGGCGGTGGCCCTCCACCT
25
1159
|
|
1131141
682
701
N/A
N/A
AGCAGCTTGCCTTGGTGTCC
29
1160
|
|
1131169
722
741
4951
4970
GTCCTGGCCAGGCCGCGGTA
42
1161
|
|
1131197
778
797
5007
5026
CGTTCCGGTAGGTGGCCTCC
28
1162
|
|
1131225
872
891
5186
5205
TTCAGCACGAAGCACCACGG
27
1163
|
|
1131253
915
934
5229
5248
GCACTGTGCCAGGTCGCAGT
36
1164
|
|
1131281
1006
1025
5320
5339
GAGGCTTCGGCGGTGCCGGC
26
1165
|
|
1131309
1087
1106
5506
5525
TGGTCAGGGAAGGCGGCTGC
8
1166
|
|
1131337
1130
1149
5549
5568
AGACTCTTGCGGAGCCGCTG
24
1167
|
|
1131365
1168
1187
5587
5606
GCGCCACCAGCCCGCCAACG
33
1168
|
|
1131393
1238
1257
5657
5676
GCGATGAGGCTGCCGGCGCA
26
1169
|
|
1131421
1340
1359
6000
6019
TCACAGCTGTGGTTACGGCG
38
1170
|
|
1131449
1431
1450
N/A
N/A
AAGGCGCAACAGAGCCAGGT
55
1171
|
|
1131477
1474
1493
6217
6236
CGTAAGGCGACAGGAGCGCG
25
1172
|
|
1131505
1559
1578
6302
6321
TCGAACTGGTGGCCCCAGCC
86
74
|
|
1131533
1595
1614
6893
6912
GCCTCCTGCAGGAAGCTGGC
53
1173
|
|
1131561
1688
1707
6986
7005
TCGAGGAACCCTGCGCAGAG
21†
1174
|
|
1131589
1775
1794
7163
7182
CCTTGCAGGGTGAGCCGGCG
30
1175
|
|
1131617
1804
1823
7192
7211
CACCACAGCCCGATCCCCAG
28
1176
|
|
1131645
1851
1870
7239
7258
GGCCAGGTAGTAGGCCACAT
63
1177
|
|
1131673
1886
1905
7274
7293
CTGAGCAATCAGGAAACGGT
16
1178
|
|
1131701
1928
1947
7316
7335
ACTCTCTCACTGCGGAATCA
7
1179
|
|
1131729
1964
1983
7352
7371
GGAATGGGACACAATCTTGC
26
1180
|
|
1131757
2018
2037
7406
7425
AAAGCACTTTATTGAGTTCC
3
165
|
|
1131785
N/A
N/A
127
146
AGAACAATCCTGGGACAATC
69
1181
|
|
1131813
N/A
N/A
211
230
ACCTTTCCACATAGGCCTCC
99
1182
|
|
1131841
N/A
N/A
314
333
TTGTCTTCTTAAGGCCCATC
49
1183
|
|
1131869
N/A
N/A
404
423
ACCGATCTGTTGCTAGTCTG
72
1184
|
|
1131897
N/A
N/A
535
554
GGAGGAGGAGCCAGGCCACT
94
1185
|
|
1131925
N/A
N/A
639
658
TCTAGTGCCCATGGGCATAA
38
1186
|
|
1131953
N/A
N/A
757
776
GAGCCCAGGTGTGATGGCGC
87
1187
|
|
1131981
N/A
N/A
895
914
AGATGCCAGAGACAAGCTCT
49
1188
|
|
1132009
N/A
N/A
1009
1028
TTTACTGTTATCAACCTGGT
38
1189
|
|
1132037
N/A
N/A
1108
1127
AGTTTCATAATAAAAGCTAA
83
1190
|
|
1132065
N/A
N/A
1524
1543
GCTATGTGCCACACTCTAGG
102
1191
|
|
1132093
N/A
N/A
1625
1644
AGTCCAGGGTTGGAATGTGC
80
1192
|
|
1132121
N/A
N/A
1920
1939
TAGCTGAGCATGAAGTTGTG
52
1193
|
|
1132149
N/A
N/A
2169
2188
AATAAGCTTTTGCCACTATT
59
1194
|
|
1132177
N/A
N/A
2286
2305
AACATAACCATCGCGAATAA
53
1195
|
|
1132205
N/A
N/A
2400
2419
CAGAATCATACATAAAACCA
45
1196
|
|
1132233
N/A
N/A
2539
2558
GCTGTGTAGTTCCTAAGCAC
70
1197
|
|
1132261
N/A
N/A
2661
2680
TGTTTCAGCTCTCCTATTGA
53
1198
|
|
1132289
N/A
N/A
2774
2793
CTCCCAAGTCAATACTTGAA
93
1199
|
|
1132317
N/A
N/A
2870
2889
TTATCTGTAAAACGGCAGTA
38
1200
|
|
1132345
N/A
N/A
2977
2996
CTTCAGTCCCAAGGCCAGGA
64
1201
|
|
1132373
N/A
N/A
3385
3404
TGAAAACACTTTCTGCTCTA
51
1202
|
|
1132401
N/A
N/A
3472
3491
GGACTTCCCCTGTACTCAAC
72
1203
|
|
1132429
N/A
N/A
3670
3689
AGTTCCTTGGACAGAAGGAC
50
1204
|
|
1132457
N/A
N/A
3836
3855
CTGTGTGTAGCACCTTTCAC
73
1205
|
|
1132485
N/A
N/A
3960
3979
TGGACTTCCAAAGGGTATTG
39
1206
|
|
1132513
N/A
N/A
4048
4067
ACTTGCCTTGTGACCTTGCT
58
1207
|
|
1132541
N/A
N/A
4248
4267
CACATCTCCTCACCTTTCTG
98
1208
|
|
1132569
N/A
N/A
4349
4368
TCTCAGGAGAGTCTAGGACC
59
1209
|
|
1132597
N/A
N/A
4581
4600
CCTGGCACACCACCCGGCCT
70
1210
|
|
1132625
N/A
N/A
4781
4800
CTGGCCTTCTGCTTGCCCCA
46
1211
|
|
1132653
N/A
N/A
5067
5086
CGGCGCACCGGCAGAAGGCG
37
1212
|
|
1132681
N/A
N/A
5430
5449
GCCGGCTGGCCGGAATCTAG
77
1213
|
|
1132709
N/A
N/A
5856
5875
CCGTGTTCCAGCTTCCTCCC
160
1214
|
|
1132737
N/A
N/A
6318
6337
GCAGTTGTGCCTACCCTCGA
49
1215
|
|
1132765
N/A
N/A
6409
6428
CATTCTGTAGGCACCCGGAA
47
1216
|
|
1132793
N/A
N/A
6530
6549
GCAAGTTCGATTCTCCCTGT
72
1217
|
|
1132821
N/A
N/A
6655
6674
CGGAAACAGAAACCCCTCCC
47
1218
|
|
1132849
N/A
N/A
6761
6780
CCTTCCTCTCTCGCAGCAAG
59
1219
|
|
1132877
N/A
N/A
7016
7035
GAGCTCACCTGGCACGCATC
85†
1220
|
|
TABLE 20
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed
|
PO/PS internucleoside linkages (Huh7, electroporation,
|
5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
FXII
SEQ ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130750
13
32
13
32
CCAGCTGCCTATCCAGGAGT
48
1221
|
|
1130778
50
69
50
69
AGGAGCAGCAGAGCCCTCAT
38
1222
|
|
1130806
97
116
N/A
N/A
AAGGTGGAATCGAAAGTGTT
67
1223
|
|
1130834
152
171
N/A
N/A
GTGAGAACGACTGTGTGCTC
20
1224
|
|
1130862
206
225
3557
3576
TGGTACAGCTGCCGGTGGTA
26
1225
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
13
183
|
|
1130891
241
260
3592
3611
GCTGAGGGCCTGGCCGGCCC
51
1226
|
|
1130919
311
330
3818
3837
ACTTTCTTGGGCTCCAAACA
49
1227
|
|
1130947
367
386
4175
4194
TGTTCACACAGGTCCCTCCT
28
1228
|
|
1130975
426
445
4234
4253
TTTCTGGCAGTGGTTTCCAG
24
1229
|
|
1131003
469
488
4414
4433
TCTTGTGGAAAAACCGGAGA
36
1230
|
|
1131031
509
528
4454
4473
CTGGCCACAGCTGCTTGCTC
46
1231
|
|
1131059
549
568
4494
4513
CAGCCGCTGGCAGTGGGCAT
71
1232
|
|
1131087
585
604
4669
4688
ATGGAGGCACGGGTTGGTGC
64
1233
|
|
1131115
624
643
4708
4727
GCACAGGCGGTGGCCCTCCA
34
1234
|
|
1131143
686
705
4915
4934
TCATAGCAGCTTGCCTTGGT
28
1235
|
|
1131171
725
744
4954
4973
GTGGTCCTGGCCAGGCCGCG
64
1236
|
|
1131199
797
816
5026
5045
CGCGCTTGCTCGGCAGTCAC
17
1237
|
|
1131227
874
893
5188
5207
GGTTCAGCACGAAGCACCAC
23
1238
|
|
1131255
917
936
5231
5250
TGGCACTGTGCCAGGTCGCA
18
1239
|
|
1131283
1008
1027
5322
5341
CTGAGGCTTCGGCGGTGCCG
21
1240
|
|
1131311
1090
1109
5509
5528
TCCTGGTCAGGGAAGGCGGC
26
1241
|
|
1131339
1132
1151
5551
5570
ACAGACTCTTGCGGAGCCGC
20
60
|
|
1131367
1204
1223
5623
5642
AGTACAGCGCGGCGATGTAG
42
1242
|
|
1131395
1240
1259
5659
5678
GGGCGATGAGGCTGCCGGCG
53
1243
|
|
1131423
1342
1361
6002
6021
GCTCACAGCTGTGGTTACGG
19
1244
|
|
1131451
1434
1453
N/A
N/A
CTGAAGGCGCAACAGAGCCA
46
1245
|
|
1131479
1493
1512
6236
6255
GGCAGGCACACCGGCTGAAC
31
1246
|
|
1131507
1561
1580
6304
6323
CCTCGAACTGGTGGCCCCAG
153
1247
|
|
1131535
1598
1617
6896
6915
TGCGCCTCCTGCAGGAAGCT
116
1248
|
|
1131563
1691
1710
6989
7008
CCCTCGAGGAACCCTGCGCA
23†
1249
|
|
1131591
1777
1796
7165
7184
TGCCTTGCAGGGTGAGCCGG
19
1250
|
|
1131619
1806
1825
7194
7213
GTCACCACAGCCCGATCCCC
37
1251
|
|
1131647
1853
1872
7241
7260
CAGGCCAGGTAGTAGGCCAC
48
1252
|
|
1131675
1893
1912
7281
7300
TGAGTCCCTGAGCAATCAGG
19
150
|
|
1131703
1932
1951
7320
7339
AGCCACTCTCTCACTGCGGA
24
1253
|
|
1131731
1985
2004
7373
7392
CGCGGAGCTGGCCGCACTGG
30
1254
|
|
1131759
2020
2039
7408
7427
TCAAAGCACTTTATTGAGTT
11
1255
|
|
1131787
N/A
N/A
133
152
CCCCCCAGAACAATCCTGGG
80
1256
|
|
1131815
N/A
N/A
217
236
GGCCTCACCTTTCCACATAG
66
1257
|
|
1131843
N/A
N/A
320
339
CCCCCGTTGTCTTCTTAAGG
47
1258
|
|
1131871
N/A
N/A
410
429
CCTGCCACCGATCTGTTGCT
124
1259
|
|
1131899
N/A
N/A
541
560
TTCCCGGGAGGAGGAGCCAG
46
1260
|
|
1131927
N/A
N/A
645
664
GATCACTCTAGTGCCCATGG
48
1261
|
|
1131955
N/A
N/A
763
782
TGGCAGGAGCCCAGGTGTGA
66
1262
|
|
1131983
N/A
N/A
901
920
GCATTCAGATGCCAGAGACA
77
1263
|
|
1132011
N/A
N/A
1015
1034
AAGAGGTTTACTGTTATCAA
55
1264
|
|
1132039
N/A
N/A
1114
1133
AAAGGTAGTTTCATAATAAA
44
1265
|
|
1132067
N/A
N/A
1530
1549
GGCCCTGCTATGTGCCACAC
93
1266
|
|
1132095
N/A
N/A
1640
1659
AGAGGAGGCTGAGGGAGTCC
52
1267
|
|
1132123
N/A
N/A
2075
2094
ACCTTTAAGATGCAAATGGG
51
1268
|
|
1132151
N/A
N/A
2175
2194
CTGCTCAATAAGCTTTTGCC
54
1269
|
|
1132179
N/A
N/A
2292
2311
TTATAGAACATAACCATCGC
68
1270
|
|
1132207
N/A
N/A
2426
2445
CATATACTAAATAAAATGTC
99
1271
|
|
1132235
N/A
N/A
2545
2564
TGAAGAGCTGTGTAGTTCCT
58
1272
|
|
1132263
N/A
N/A
2667
2686
GCTGCTTGTTTCAGCTCTCC
55
1273
|
|
1132291
N/A
N/A
2780
2799
TTGTAACTCCCAAGTCAATA
78
1274
|
|
1132319
N/A
N/A
2893
2912
CTCTGATCTGAGCCTTGGTT
95
1275
|
|
1132347
N/A
N/A
2983
3002
GTAAGTCTTCAGTCCCAAGG
86
1276
|
|
1132375
N/A
N/A
3391
3410
GTGCATTGAAAACACTTTCT
41
1277
|
|
1132403
N/A
N/A
3478
3497
CTCTGAGGACTTCCCCTGTA
166
1278
|
|
1132431
N/A
N/A
3676
3695
AAGCAGAGTTCCTTGGACAG
57
1279
|
|
1132459
N/A
N/A
3842
3861
CAGAGGCTGTGTGTAGCACC
76
1280
|
|
1132487
N/A
N/A
3966
3985
TCCCTCTGGACTTCCAAAGG
47
1281
|
|
1132515
N/A
N/A
4054
4073
GAACCTACTTGCCTTGTGAC
59
1282
|
|
1132543
N/A
N/A
4254
4273
GTCCTCCACATCTCCTCACC
47
1283
|
|
1132571
N/A
N/A
4355
4374
TAGTGGTCTCAGGAGAGTCT
92
1284
|
|
1132599
N/A
N/A
4587
4606
CTCCTTCCTGGCACACCACC
38
1285
|
|
1132627
N/A
N/A
4803
4822
GCAAGCCCGTCCCACCTGGG
47
1286
|
|
1132655
N/A
N/A
5073
5092
CCCACGCGGCGCACCGGCAG
48
1287
|
|
1132683
N/A
N/A
5439
5458
GAGCCCGCGGCCGGCTGGCC
68
1288
|
|
1132711
N/A
N/A
5862
5881
CCAATCCCGTGTTCCAGCTT
37
1289
|
|
1132739
N/A
N/A
6324
6343
CCCCTAGCAGTTGTGCCTAC
138
1290
|
|
1132767
N/A
N/A
6415
6434
GCCACCCATTCTGTAGGCAC
59
1291
|
|
1132795
N/A
N/A
6536
6555
CCAAGAGCAAGTTCGATTCT
50
1292
|
|
1132823
N/A
N/A
6661
6680
GCGGAGCGGAAACAGAAACC
88
1293
|
|
1132851
N/A
N/A
6767
6786
TCATGCCCTTCCTCTCTCGC
39
1294
|
|
1132879
N/A
N/A
7025
7044
CCGGGCTAAGAGCTCACCTG
59†
1295
|
|
TABLE 21
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed
|
PO/PS internucleoside linkages (Huh7, electroporation,
|
5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
FXII
SEQ ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130752
15
34
15
34
GTCCAGCTGCCTATCCAGGA
49
1296
|
|
1130780
57
76
57
76
GAACCCCAGGAGCAGCAGAG
72
1297
|
|
1130808
103
122
N/A
N/A
CTTCCCAAGGTGGAATCGAA
62
1298
|
|
1130836
154
173
N/A
N/A
CAGTGAGAACGACTGTGTGC
41
1299
|
|
1130864
208
227
3559
3578
TGTGGTACAGCTGCCGGTGG
42
1300
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
21
183
|
|
1130893
254
273
N/A
N/A
GTAGCACACCAGGGCTGAGG
54
1301
|
|
1130921
313
332
3820
3839
TCACTTTCTTGGGCTCCAAA
41
1302
|
|
1130949
371
390
4179
4198
GGCATGTTCACACAGGTCCC
42
1303
|
|
1130977
428
447
N/A
N/A
TCTTTCTGGCAGTGGTTTCC
48
1304
|
|
1131005
471
490
4416
4435
ATTCTTGTGGAAAAACCGGA
39
1305
|
|
1131033
511
530
4456
4475
ATCTGGCCACAGCTGCTTGC
53
1306
|
|
1131061
551
570
4496
4515
GCCAGCCGCTGGCAGTGGGC
58
1307
|
|
1131089
588
607
4672
4691
CCCATGGAGGCACGGGTTGG
40
1308
|
|
1131117
627
646
4711
4730
GTGGCACAGGCGGTGGCCCT
71
1309
|
|
1131145
690
709
4919
4938
GCCATCATAGCAGCTTGCCT
36
1310
|
|
1131173
745
764
4974
4993
GACAGGGCGCACCCGAGAGC
52
1311
|
|
1131201
799
818
5028
5047
TCCGCGCTTGCTCGGCAGTC
74
1312
|
|
1131229
882
901
5196
5215
CCGGTCGCGGTTCAGCACGA
46
1313
|
|
1131257
920
939
5234
5253
GTCTGGCACTGTGCCAGGTC
74
1314
|
|
1131285
1011
1030
5325
5344
GGGCTGAGGCTTCGGCGGTG
31
1315
|
|
1131313
1092
1111
5511
5530
GTTCCTGGTCAGGGAAGGCG
52
1316
|
|
1131341
1134
1153
5553
5572
AGACAGACTCTTGCGGAGCC
37
1317
|
|
1131369
1207
1226
5626
5645
CCCAGTACAGCGCGGCGATG
32
1318
|
|
1131397
1281
1300
5700
5719
CCGGTCCTGCAGGCAGTGAG
59
1319
|
|
1131425
1345
1364
6005
6024
ACGGCTCACAGCTGTGGTTA
49
1320
|
|
1131453
1437
1456
6180
6199
CTCCTGAAGGCGCAACAGAG
66
1321
|
|
1131481
1496
1515
6239
6258
CTTGGCAGGCACACCGGCTG
24
1322
|
|
1131509
1564
1583
N/A
N/A
CCCCCTCGAACTGGTGGCCC
76
1323
|
|
1131537
1603
1622
6901
6920
GTACCTGCGCCTCCTGCAGG
126
1324
|
|
1131565
1696
1715
6994
7013
TGCCGCCCTCGAGGAACCCT
8†
1325
|
|
1131593
1780
1799
7168
7187
TGATGCCTTGCAGGGTGAGC
17
1326
|
|
1131621
1808
1827
7196
7215
CGGTCACCACAGCCCGATCC
35
1327
|
|
1131649
1856
1875
7244
7263
ATCCAGGCCAGGTAGTAGGC
70
1328
|
|
1131677
1895
1914
7283
7302
GATGAGTCCCTGAGCAATCA
35
1329
|
|
1131705
1935
1954
7323
7342
CCCAGCCACTCTCTCACTGC
37
1330
|
|
1131733
1989
2008
7377
7396
CTGGCGCGGAGCTGGCCGCA
49
1331
|
|
1131761
2023
2042
7411
7430
TTTTCAAAGCACTTTATTGA
28
1332
|
|
1131789
N/A
N/A
139
158
TAGCGACCCCCCAGAACAAT
101
1333
|
|
1131817
N/A
N/A
223
242
CGGGCTGGCCTCACCTTTCC
74
1334
|
|
1131845
N/A
N/A
326
345
TACCTCCCCCCGTTGTCTTC
105
1335
|
|
1131873
N/A
N/A
416
435
TCATAGCCTGCCACCGATCT
77
1336
|
|
1131901
N/A
N/A
560
579
ACACATCCCCACCCAAGGGT
61
1337
|
|
1131929
N/A
N/A
651
670
GCTCACGATCACTCTAGTGC
44
1338
|
|
1131957
N/A
N/A
770
789
GGAAGGCTGGCAGGAGCCCA
81
1339
|
|
1131985
N/A
N/A
910
929
GCTTCAGAGGCATTCAGATG
84
1340
|
|
1132013
N/A
N/A
1021
1040
AACCCTAAGAGGTTTACTGT
105
1341
|
|
1132041
N/A
N/A
1120
1139
CATCTAAAAGGTAGTTTCAT
90
1342
|
|
1132069
N/A
N/A
1536
1555
TTCACAGGCCCTGCTATGTG
101
1343
|
|
1132097
N/A
N/A
1667
1686
GGAGAAACTTTTGGGTGTGG
86
1344
|
|
1132125
N/A
N/A
2081
2100
AGATGGACCTTTAAGATGCA
79
1345
|
|
1132153
N/A
N/A
2181
2200
AAGTACCTGCTCAATAAGCT
61
1346
|
|
1132181
N/A
N/A
2299
2318
TGTGATTTTATAGAACATAA
71
1347
|
|
1132209
N/A
N/A
2437
2456
AATCAGCAACACATATACTA
73
1348
|
|
1132237
N/A
N/A
2551
2570
TCCTGCTGAAGAGCTGTGTA
66
1349
|
|
1132265
N/A
N/A
2673
2692
GCTGCTGCTGCTTGTTTCAG
42
1350
|
|
1132293
N/A
N/A
2786
2805
TTTTATTTGTAACTCCCAAG
100
1351
|
|
1132321
N/A
N/A
2899
2918
TAACCACTCTGATCTGAGCC
80
1352
|
|
1132349
N/A
N/A
2989
3008
CCTTGGGTAAGTCTTCAGTC
107
1353
|
|
1132377
N/A
N/A
3397
3416
GTCGCTGTGCATTGAAAACA
98
1354
|
|
1132405
N/A
N/A
3484
3503
CACACTCTCTGAGGACTTCC
83
1355
|
|
1132433
N/A
N/A
3682
3701
CTCTCCAAGCAGAGTTCCTT
89
1356
|
|
1132461
N/A
N/A
3848
3867
CCACCCCAGAGGCTGTGTGT
90
1357
|
|
1132489
N/A
N/A
3972
3991
GAACTCTCCCTCTGGACTTC
61
1358
|
|
1132517
N/A
N/A
4060
4079
CTTCTTGAACCTACTTGCCT
66
1359
|
|
1132545
N/A
N/A
4260
4279
GCCCAGGTCCTCCACATCTC
60
1360
|
|
1132573
N/A
N/A
4361
4380
GAGGGATAGTGGTCTCAGGA
76
1361
|
|
1132601
N/A
N/A
4593
4612
AGAGCTCTCCTTCCTGGCAC
60
1362
|
|
1132629
N/A
N/A
4809
4828
TTCCTGGCAAGCCCGTCCCA
50
1363
|
|
1132657
N/A
N/A
5079
5098
CCCAGCCCCACGCGGCGCAC
105
1364
|
|
1132685
N/A
N/A
5445
5464
GACGGAGAGCCCGCGGCCGG
109
1365
|
|
1132713
N/A
N/A
5868
5887
CGAACCCCAATCCCGTGTTC
81
1366
|
|
1132741
N/A
N/A
6330
6349
CCCCTGCCCCTAGCAGTTGT
111
1367
|
|
1132769
N/A
N/A
6436
6455
CTCACAACCCATCAGGTCAG
97
1368
|
|
1132797
N/A
N/A
6542
6561
AGGGAACCAAGAGCAAGTTC
26
1369
|
|
1132825
N/A
N/A
6667
6686
GAATGGGCGGAGCGGAAACA
73
1370
|
|
1132853
N/A
N/A
6773
6792
ACCCACTCATGCCCTTCCTC
42
1371
|
|
1132881
N/A
N/A
7031
7050
CGCCAACCGGGCTAAGAGCT
82†
1372
|
|
TABLE 22
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed
|
PO/PS internucleoside linkages (Huh7, electroporation,
|
5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
FXII
SEQ ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130753
16
35
16
35
GGTCCAGCTGCCTATCCAGG
37
1373
|
|
1130781
60
79
60
79
CAGGAACCCCAGGAGCAGCA
65
1374
|
|
1130809
104
123
N/A
N/A
GCTTCCCAAGGTGGAATCGA
67
1375
|
|
1130837
156
175
N/A
N/A
GACAGTGAGAACGACTGTGT
43
1376
|
|
1130865
210
229
3561
3580
TTTGTGGTACAGCTGCCGGT
52
1377
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
20
183
|
|
1130894
257
276
N/A
N/A
GTGGTAGCACACCAGGGCTG
19
1378
|
|
1130922
314
333
3821
3840
TTCACTTTCTTGGGCTCCAA
36
1379
|
|
1130950
372
391
4180
4199
TGGCATGTTCACACAGGTCC
42
1380
|
|
1130978
429
448
N/A
N/A
CTCTTTCTGGCAGTGGTTTC
45
1381
|
|
1131006
472
491
4417
4436
CATTCTTGTGGAAAAACCGG
22
1382
|
|
1131034
512
531
4457
4476
CATCTGGCCACAGCTGCTTG
42
1383
|
|
1131062
552
571
4497
4516
GGCCAGCCGCTGGCAGTGGG
87
1384
|
|
1131090
589
608
4673
4692
CCCCATGGAGGCACGGGTTG
36
1385
|
|
1131118
628
647
4712
4731
AGTGGCACAGGCGGTGGCCC
32
1386
|
|
1131146
691
710
4920
4939
GGCCATCATAGCAGCTTGCC
27
1387
|
|
1131174
747
766
4976
4995
CTGACAGGGCGCACCCGAGA
49
1388
|
|
1131202
800
819
5029
5048
TTCCGCGCTTGCTCGGCAGT
35
1389
|
|
1131230
883
902
5197
5216
GCCGGTCGCGGTTCAGCACG
42
1390
|
|
1131258
921
940
5235
5254
GGTCTGGCACTGTGCCAGGT
34
1391
|
|
1131286
1012
1031
5326
5345
TGGGCTGAGGCTTCGGCGGT
45
1392
|
|
1131314
1093
1112
5512
5531
CGTTCCTGGTCAGGGAAGGC
15
1393
|
|
1131342
1135
1154
5554
5573
AAGACAGACTCTTGCGGAGC
29
1394
|
|
1131370
1209
1228
5628
5647
GCCCCAGTACAGCGCGGCGA
53
1395
|
|
1131398
1282
1301
5701
5720
GCCGGTCCTGCAGGCAGTGA
48
1396
|
|
1131426
1346
1365
6006
6025
CACGGCTCACAGCTGTGGTT
28
1397
|
|
1131454
1439
1458
6182
6201
TCCTCCTGAAGGCGCAACAG
33
1398
|
|
1131482
1497
1516
6240
6259
GCTTGGCAGGCACACCGGCT
41
1399
|
|
1131510
1565
1584
N/A
N/A
GCCCCCTCGAACTGGTGGCC
49
1400
|
|
1131538
1604
1623
6902
6921
GGTACCTGCGCCTCCTGCAG
154
1401
|
|
1131566
1699
1718
6997
7016
CGGTGCCGCCCTCGAGGAAC
6†
1402
|
|
1131594
1781
1800
7169
7188
ATGATGCCTTGCAGGGTGAG
29
1403
|
|
1131622
1810
1829
7198
7217
TGCGGTCACCACAGCCCGAT
37
1404
|
|
1131650
1857
1876
7245
7264
GATCCAGGCCAGGTAGTAGG
29
1405
|
|
1131678
1896
1915
7284
7303
AGATGAGTCCCTGAGCAATC
32
1406
|
|
1131706
1938
1957
7326
7345
TGCCCCAGCCACTCTCTCAC
25
1407
|
|
1131734
1990
2009
7378
7397
CCTGGCGCGGAGCTGGCCGC
44
1408
|
|
1131762
2024
2043
7412
7431
ATTTTCAAAGCACTTTATTG
32
1409
|
|
1131790
N/A
N/A
142
161
TGATAGCGACCCCCCAGAAC
80
1410
|
|
1131818
N/A
N/A
229
248
GCCTTCCGGGCTGGCCTCAC
91
1411
|
|
1131846
N/A
N/A
329
348
TTCTACCTCCCCCCGTTGTC
106
1412
|
|
1131874
N/A
N/A
419
438
AAGTCATAGCCTGCCACCGA
105
1413
|
|
1131902
N/A
N/A
563
582
CATACACATCCCCACCCAAG
85
1414
|
|
1131930
N/A
N/A
654
673
ACAGCTCACGATCACTCTAG
67
1415
|
|
1131958
N/A
N/A
773
792
TGAGGAAGGCTGGCAGGAGC
76
1416
|
|
1131986
N/A
N/A
913
932
TGGGCTTCAGAGGCATTCAG
50
1417
|
|
1132014
N/A
N/A
1024
1043
AACAACCCTAAGAGGTTTAC
62
1418
|
|
1132042
N/A
N/A
1123
1142
CTTCATCTAAAAGGTAGTTT
48
1419
|
|
1132070
N/A
N/A
1539
1558
TGGTTCACAGGCCCTGCTAT
66
1420
|
|
1132098
N/A
N/A
1670
1689
AGAGGAGAAACTTTTGGGTG
68
1421
|
|
1132126
N/A
N/A
2084
2103
CTGAGATGGACCTTTAAGAT
54
1422
|
|
1132154
N/A
N/A
2188
2207
TGGCACAAAGTACCTGCTCA
53
1423
|
|
1132182
N/A
N/A
2302
2321
CACTGTGATTTTATAGAACA
65
1424
|
|
1132210
N/A
N/A
2440
2459
ATGAATCAGCAACACATATA
58
1425
|
|
1132238
N/A
N/A
2554
2573
GCCTCCTGCTGAAGAGCTGT
61
1426
|
|
1132266
N/A
N/A
2676
2695
CACGCTGCTGCTGCTTGTTT
75
1427
|
|
1132294
N/A
N/A
2789
2808
GAATTTTATTTGTAACTCCC
84
1428
|
|
1132322
N/A
N/A
2902
2921
TGTTAACCACTCTGATCTGA
59
1429
|
|
1132350
N/A
N/A
2992
3011
TGACCTTGGGTAAGTCTTCA
93
1430
|
|
1132378
N/A
N/A
3400
3419
AAGGTCGCTGTGCATTGAAA
84
1431
|
|
1132406
N/A
N/A
3487
3506
CAACACACTCTCTGAGGACT
55
1432
|
|
1132434
N/A
N/A
3685
3704
CCTCTCTCCAAGCAGAGTTC
49
1433
|
|
1132462
N/A
N/A
3851
3870
AGGCCACCCCAGAGGCTGTG
38
1434
|
|
1132490
N/A
N/A
3975
3994
CCAGAACTCTCCCTCTGGAC
74
1435
|
|
1132518
N/A
N/A
4063
4082
GCCCTTCTTGAACCTACTTG
73
1436
|
|
1132546
N/A
N/A
4263
4282
CCCGCCCAGGTCCTCCACAT
59
1437
|
|
1132574
N/A
N/A
4364
4383
AAAGAGGGATAGTGGTCTCA
74
1438
|
|
1132602
N/A
N/A
4596
4615
CAGAGAGCTCTCCTTCCTGG
81
1439
|
|
1132630
N/A
N/A
4812
4831
TCCTTCCTGGCAAGCCCGTC
40
1440
|
|
1132658
N/A
N/A
5082
5101
TCACCCAGCCCCACGCGGCG
44
1441
|
|
1132686
N/A
N/A
5448
5467
GAGGACGGAGAGCCCGCGGC
75
1442
|
|
1132714
N/A
N/A
5871
5890
TCCCGAACCCCAATCCCGTG
83
1443
|
|
1132742
N/A
N/A
6333
6352
CTACCCCTGCCCCTAGCAGT
70
1444
|
|
1132770
N/A
N/A
6439
6458
ATTCTCACAACCCATCAGGT
103
1445
|
|
1132798
N/A
N/A
6548
6567
GCCCAGAGGGAACCAAGAGC
85
1446
|
|
1132826
N/A
N/A
6670
6689
TTTGAATGGGCGGAGCGGAA
86
1447
|
|
1132854
N/A
N/A
6776
6795
TAAACCCACTCATGCCCTTC
87
1448
|
|
1132882
N/A
N/A
7057
7076
TTGTGCCTGACGGCCTCGGG
84†
1449
|
|
TABLE 23
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed
|
PO/PS internucleoside linkages (Huh7, electroporation,
|
5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
FXII
SEQ ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130751
14
33
14
33
TCCAGCTGCCTATCCAGGAG
65
1450
|
|
1130779
51
70
51
70
CAGGAGCAGCAGAGCCCTCA
39
1451
|
|
1130807
99
118
N/A
N/A
CCAAGGTGGAATCGAAAGTG
37
1452
|
|
1130835
153
172
N/A
N/A
AGTGAGAACGACTGTGTGCT
29
1453
|
|
1130863
207
226
3558
3577
GTGGTACAGCTGCCGGTGGT
23
1454
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
8
183
|
|
1130892
242
261
3593
3612
GGCTGAGGGCCTGGCCGGCC
54
1455
|
|
1130920
312
331
3819
3838
CACTTTCTTGGGCTCCAAAC
17
1456
|
|
1130948
370
389
4178
4197
GCATGTTCACACAGGTCCCT
36
1457
|
|
1130976
427
446
4235
4254
CTTTCTGGCAGTGGTTTCCA
25
1458
|
|
1131004
470
489
4415
4434
TTCTTGTGGAAAAACCGGAG
28
1459
|
|
1131032
510
529
4455
4474
TCTGGCCACAGCTGCTTGCT
26
1460
|
|
1131060
550
569
4495
4514
CCAGCCGCTGGCAGTGGGCA
53
1461
|
|
1131088
586
605
4670
4689
CATGGAGGCACGGGTTGGTG
89
1462
|
|
1131116
625
644
4709
4728
GGCACAGGCGGTGGCCCTCC
47
1463
|
|
1131144
688
707
4917
4936
CATCATAGCAGCTTGCCTTG
40
1464
|
|
1131172
744
763
4973
4992
ACAGGGCGCACCCGAGAGCG
50
1465
|
|
1131200
798
817
5027
5046
CCGCGCTTGCTCGGCAGTCA
31
1466
|
|
1131228
876
895
5190
5209
GCGGTTCAGCACGAAGCACC
47
1467
|
|
1131256
919
938
5233
5252
TCTGGCACTGTGCCAGGTCG
43
1468
|
|
1131284
1010
1029
5324
5343
GGCTGAGGCTTCGGCGGTGC
27
1469
|
|
1131312
1091
1110
5510
5529
TTCCTGGTCAGGGAAGGCGG
35
1470
|
|
1131340
1133
1152
5552
5571
GACAGACTCTTGCGGAGCCG
19
1471
|
|
1131368
1206
1225
5625
5644
CCAGTACAGCGCGGCGATGT
17
1472
|
|
1131396
1280
1299
5699
5718
CGGTCCTGCAGGCAGTGAGC
26
1473
|
|
1131424
1344
1363
6004
6023
CGGCTCACAGCTGTGGTTAC
19
1474
|
|
1131452
1436
1455
6179
6198
TCCTGAAGGCGCAACAGAGC
39
1475
|
|
1131480
1494
1513
6237
6256
TGGCAGGCACACCGGCTGAA
22
1476
|
|
1131508
1563
1582
N/A
N/A
CCCCTCGAACTGGTGGCCCC
132
1477
|
|
1131536
1601
1620
6899
6918
ACCTGCGCCTCCTGCAGGAA
108
1478
|
|
1131564
1694
1713
6992
7011
CCGCCCTCGAGGAACCCTGC
10†
1479
|
|
1131592
1778
1797
7166
7185
ATGCCTTGCAGGGTGAGCCG
24
1480
|
|
1131620
1807
1826
7195
7214
GGTCACCACAGCCCGATCCC
23
1481
|
|
1131648
1855
1874
7243
7262
TCCAGGCCAGGTAGTAGGCC
81
1482
|
|
1131676
1894
1913
7282
7301
ATGAGTCCCTGAGCAATCAG
44
1483
|
|
1131704
1933
1952
7321
7340
CAGCCACTCTCTCACTGCGG
11
1484
|
|
1131732
1986
2005
7374
7393
GCGCGGAGCTGGCCGCACTG
24
1485
|
|
1131760
2021
2040
7409
7428
TTCAAAGCACTTTATTGAGT
6
1486
|
|
1131788
N/A
N/A
136
155
CGACCCCCCAGAACAATCCT
54
1487
|
|
1131816
N/A
N/A
220
239
GCTGGCCTCACCTTTCCACA
69
1488
|
|
1131844
N/A
N/A
323
342
CTCCCCCCGTTGTCTTCTTA
87
1489
|
|
1131872
N/A
N/A
413
432
TAGCCTGCCACCGATCTGTT
59
1490
|
|
1131900
N/A
N/A
557
576
CATCCCCACCCAAGGGTTCC
80
1491
|
|
1131928
N/A
N/A
648
667
CACGATCACTCTAGTGCCCA
67
1492
|
|
1131956
N/A
N/A
767
786
AGGCTGGCAGGAGCCCAGGT
61
1493
|
|
1131984
N/A
N/A
904
923
GAGGCATTCAGATGCCAGAG
101
1494
|
|
1132012
N/A
N/A
1018
1037
CCTAAGAGGTTTACTGTTAT
46
1495
|
|
1132040
N/A
N/A
1117
1136
CTAAAAGGTAGTTTCATAAT
70
1496
|
|
1132068
N/A
N/A
1533
1552
ACAGGCCCTGCTATGTGCCA
67
1497
|
|
1132096
N/A
N/A
1647
1666
GGTGAAGAGAGGAGGCTGAG
68
1498
|
|
1132124
N/A
N/A
2078
2097
TGGACCTTTAAGATGCAAAT
53
1499
|
|
1132152
N/A
N/A
2178
2197
TACCTGCTCAATAAGCTTTT
54
1500
|
|
1132180
N/A
N/A
2295
2314
ATTTTATAGAACATAACCAT
125
1501
|
|
1132208
N/A
N/A
2430
2449
AACACATATACTAAATAAAA
88
1502
|
|
1132236
N/A
N/A
2548
2567
TGCTGAAGAGCTGTGTAGTT
77
1503
|
|
1132264
N/A
N/A
2670
2689
GCTGCTGCTTGTTTCAGCTC
72
1504
|
|
1132292
N/A
N/A
2783
2802
TATTTGTAACTCCCAAGTCA
66
1505
|
|
1132320
N/A
N/A
2896
2915
CCACTCTGATCTGAGCCTTG
87
1506
|
|
1132348
N/A
N/A
2986
3005
TGGGTAAGTCTTCAGTCCCA
64
1507
|
|
1132376
N/A
N/A
3394
3413
GCTGTGCATTGAAAACACTT
32
1508
|
|
1132404
N/A
N/A
3481
3500
ACTCTCTGAGGACTTCCCCT
48
1509
|
|
1132432
N/A
N/A
3679
3698
TCCAAGCAGAGTTCCTTGGA
102
1510
|
|
1132460
N/A
N/A
3845
3864
CCCCAGAGGCTGTGTGTAGC
80
1511
|
|
1132488
N/A
N/A
3969
3988
CTCTCCCTCTGGACTTCCAA
52
1512
|
|
1132516
N/A
N/A
4057
4076
CTTGAACCTACTTGCCTTGT
53
1513
|
|
1132544
N/A
N/A
4257
4276
CAGGTCCTCCACATCTCCTC
57
1514
|
|
1132572
N/A
N/A
4358
4377
GGATAGTGGTCTCAGGAGAG
82
1515
|
|
1132600
N/A
N/A
4590
4609
GCTCTCCTTCCTGGCACACC
73
1516
|
|
1132628
N/A
N/A
4806
4825
CTGGCAAGCCCGTCCCACCT
52
1517
|
|
1132656
N/A
N/A
5076
5095
AGCCCCACGCGGCGCACCGG
60
1518
|
|
1132684
N/A
N/A
5442
5461
GGAGAGCCCGCGGCCGGCTG
72
1519
|
|
1132712
N/A
N/A
5865
5884
ACCCCAATCCCGTGTTCCAG
64
1520
|
|
1132740
N/A
N/A
6327
6346
CTGCCCCTAGCAGTTGTGCC
93
1521
|
|
1132768
N/A
N/A
6433
6452
ACAACCCATCAGGTCAGCGC
109
1522
|
|
1132796
N/A
N/A
6539
6558
GAACCAAGAGCAAGTTCGAT
67
1523
|
|
1132824
N/A
N/A
6664
6683
TGGGCGGAGCGGAAACAGAA
46
1524
|
|
1132852
N/A
N/A
6770
6789
CACTCATGCCCTTCCTCTCT
39
1525
|
|
1132880
N/A
N/A
7028
7047
CAACCGGGCTAAGAGCTCAC
56†
1526
|
|
TABLE 24
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed
|
PO/PS internucleoside linkages (Huh7, electroporation,
|
5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130754
18
37
18
37
TTGGTCCAGCTGCCTATCCA
28
1527
|
|
1130782
65
84
65
84
ACCAGCAGGAACCCCAGGAG
94
1528
|
|
1130810
106
125
473
492
GGGCTTCCCAAGGTGGAATC
19
1529
|
|
1130838
157
176
N/A
N/A
TGACAGTGAGAACGACTGTG
33
1530
|
|
1130866
211
230
3562
3581
ATTTGTGGTACAGCTGCCGG
23
1531
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
11
183
|
|
1130895
258
277
N/A
N/A
GGTGGTAGCACACCAGGGCT
48
1532
|
|
1130923
316
335
3823
3842
CTTTCACTTTCTTGGGCTCC
19
1533
|
|
1130951
374
393
4182
4201
CTTGGCATGTTCACACAGGT
26
1534
|
|
1130979
430
449
N/A
N/A
TCTCTTTCTGGCAGTGGTTT
17
1535
|
|
1131007
473
492
4418
4437
TCATTCTTGTGGAAAAACCG
18
1536
|
|
1131035
513
532
4458
4477
GCATCTGGCCACAGCTGCTT
38
1537
|
|
1131063
553
572
4498
4517
TGGCCAGCCGCTGGCAGTGG
67
1538
|
|
1131091
590
609
4674
4693
CCCCCATGGAGGCACGGGTT
42
208
|
|
1131119
630
649
4714
4733
GCAGTGGCACAGGCGGTGGC
30
1539
|
|
1131147
693
712
4922
4941
GCGGCCATCATAGCAGCTTG
46
1540
|
|
1131175
748
767
4977
4996
GCTGACAGGGCGCACCCGAG
32
1541
|
|
1131203
801
820
5030
5049
GTTCCGCGCTTGCTCGGCAG
30
1542
|
|
1131231
884
903
5198
5217
AGCCGGTCGCGGTTCAGCAC
25
1543
|
|
1131259
922
941
5236
5255
GGGTCTGGCACTGTGCCAGG
22
1544
|
|
1131287
1013
1032
5327
5346
GTGGGCTGAGGCTTCGGCGG
49
1545
|
|
1131315
1094
1113
5513
5532
CCGTTCCTGGTCAGGGAAGG
46
1546
|
|
1131343
1136
1155
5555
5574
GAAGACAGACTCTTGCGGAG
38
1547
|
|
1131371
1210
1229
5629
5648
GGCCCCAGTACAGCGCGGCG
54
1548
|
|
1131399
1284
1303
N/A
N/A
GGGCCGGTCCTGCAGGCAGT
64
1549
|
|
1131427
1347
1366
6007
6026
GCACGGCTCACAGCTGTGGT
50
1550
|
|
1131455
1440
1459
6183
6202
ATCCTCCTGAAGGCGCAACA
29
1551
|
|
1131483
1499
1518
6242
6261
CCGCTTGGCAGGCACACCGG
29
1552
|
|
1131511
1566
1585
N/A
N/A
CGCCCCCTCGAACTGGTGGC
126
1553
|
|
1131539
1605
1624
6903
6922
CGGTACCTGCGCCTCCTGCA
79
1554
|
|
1131567
1700
1719
6998
7017
TCGGTGCCGCCCTCGAGGAA
5†
1555
|
|
1131595
1782
1801
7170
7189
GATGATGCCTTGCAGGGTGA
15
1556
|
|
1131623
1811
1830
7199
7218
TTGCGGTCACCACAGCCCGA
32
1557
|
|
1131651
1858
1877
7246
7265
GGATCCAGGCCAGGTAGTAG
64
1558
|
|
1131679
1897
1916
7285
7304
AAGATGAGTCCCTGAGCAAT
30
1559
|
|
1131707
1939
1958
7327
7346
ATGCCCCAGCCACTCTCTCA
24
1560
|
|
1131735
1991
2010
7379
7398
TCCTGGCGCGGAGCTGGCCG
25
1561
|
|
1131763
2026
2045
7414
7433
GCATTTTCAAAGCACTTTAT
9
1562
|
|
1131791
N/A
N/A
145
164
CTGTGATAGCGACCCCCCAG
57
1563
|
|
1131819
N/A
N/A
241
260
TCCTCTGCCTGGGCCTTCCG
83
1564
|
|
1131847
N/A
N/A
332
351
CCTTTCTACCTCCCCCCGTT
78
1565
|
|
1131875
N/A
N/A
422
441
TATAAGTCATAGCCTGCCAC
61
1566
|
|
1131903
N/A
N/A
566
585
CACCATACACATCCCCACCC
74
1567
|
|
1131931
N/A
N/A
657
676
CACACAGCTCACGATCACTC
90
1568
|
|
1131959
N/A
N/A
776
795
AACTGAGGAAGGCTGGCAGG
59
1569
|
|
1131987
N/A
N/A
919
938
TAAAGCTGGGCTTCAGAGGC
73
1570
|
|
1132015
N/A
N/A
1027
1046
CTCAACAACCCTAAGAGGTT
122
1571
|
|
1132043
N/A
N/A
1126
1145
ACCCTTCATCTAAAAGGTAG
42
1572
|
|
1132071
N/A
N/A
1542
1561
ATCTGGTTCACAGGCCCTGC
44
1573
|
|
1132099
N/A
N/A
1673
1692
GAGAGAGGAGAAACTTTTGG
51
1574
|
|
1132127
N/A
N/A
2087
2106
GATCTGAGATGGACCTTTAA
68
1575
|
|
1132155
N/A
N/A
2191
2210
GTCTGGCACAAAGTACCTGC
66
1576
|
|
1132183
N/A
N/A
2308
2327
AATGTTCACTGTGATTTTAT
86
1577
|
|
1132211
N/A
N/A
2443
2462
TTGATGAATCAGCAACACAT
86
1578
|
|
1132239
N/A
N/A
2557
2576
TGAGCCTCCTGCTGAAGAGC
64
1579
|
|
1132267
N/A
N/A
2679
2698
CGTCACGCTGCTGCTGCTTG
92
1580
|
|
1132295
N/A
N/A
2792
2811
GCTGAATTTTATTTGTAACT
101
1581
|
|
1132323
N/A
N/A
2905
2924
CACTGTTAACCACTCTGATC
49
1582
|
|
1132351
N/A
N/A
3009
3028
ACAACCTGCTAGCTGTGTGA
65
1583
|
|
1132379
N/A
N/A
3403
3422
AAAAAGGTCGCTGTGCATTG
55
1584
|
|
1132407
N/A
N/A
3490
3509
GGACAACACACTCTCTGAGG
54
1585
|
|
1132435
N/A
N/A
3688
3707
TCCCCTCTCTCCAAGCAGAG
53
1586
|
|
1132463
N/A
N/A
3855
3874
CCCCAGGCCACCCCAGAGGC
67
1587
|
|
1132491
N/A
N/A
3978
3997
TTCCCAGAACTCTCCCTCTG
59
1588
|
|
1132519
N/A
N/A
4066
4085
AAGGCCCTTCTTGAACCTAC
50
1589
|
|
1132547
N/A
N/A
4266
4285
CACCCCGCCCAGGTCCTCCA
99
1590
|
|
1132575
N/A
N/A
4387
4406
GCTCAAAGCACTTCTCTGGG
25
1591
|
|
1132603
N/A
N/A
4599
4618
CCCCAGAGAGCTCTCCTTCC
76
1592
|
|
1132631
N/A
N/A
4816
4835
CTCCTCCTTCCTGGCAAGCC
43
1593
|
|
1132659
N/A
N/A
5085
5104
GGGTCACCCAGCCCCACGCG
65
1594
|
|
1132687
N/A
N/A
5451
5470
GCTGAGGACGGAGAGCCCGC
88
1595
|
|
1132715
N/A
N/A
5874
5893
TGCTCCCGAACCCCAATCCC
43
1596
|
|
1132743
N/A
N/A
6336
6355
CCCCTACCCCTGCCCCTAGC
65
1597
|
|
1132771
N/A
N/A
6442
6461
CACATTCTCACAACCCATCA
72
1598
|
|
1132799
N/A
N/A
6566
6585
TGGCCTTTGCAGCCCGGCGC
84
1599
|
|
1132827
N/A
N/A
6673
6692
GGATTTGAATGGGCGGAGCG
49
1600
|
|
1132855
N/A
N/A
6779
6798
TTGTAAACCCACTCATGCCC
52
1601
|
|
1132883
N/A
N/A
7060
7079
GATTTGTGCCTGACGGCCTC
42†
1602
|
|
TABLE 25
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed
|
PO/PS internucleoside linkages (Huh7, electroporation,
|
5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130783
66
85
66
85
CACCAGCAGGAACCCCAGGA
57
1603
|
|
1130811
125
144
492
511
TTGTACTTATGCTCCTTGGG
30
1604
|
|
1130839
158
177
N/A
N/A
GTGACAGTGAGAACGACTGT
14
1605
|
|
1130867
212
231
3563
3582
CATTTGTGGTACAGCTGCCG
22
1606
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
17
183
|
|
1130896
278
297
3785
3804
TGGTCCTGATCAAAGTTGGG
44
1607
|
|
1130924
317
336
N/A
N/A
TCTTTCACTTTCTTGGGCTC
30
1608
|
|
1130952
375
394
4183
4202
GCTTGGCATGTTCACACAGG
15
1609
|
|
1130980
431
450
N/A
N/A
TTCTCTTTCTGGCAGTGGTT
44
1610
|
|
1131008
474
493
4419
4438
CTCATTCTTGTGGAAAAACC
25
1611
|
|
1131036
514
533
4459
4478
GGCATCTGGCCACAGCTGCT
20
1612
|
|
1131064
554
573
4499
4518
CTGGCCAGCCGCTGGCAGTG
102
1613
|
|
1131092
591
610
4675
4694
ACCCCCATGGAGGCACGGGT
45
1614
|
|
1131120
631
650
4715
4734
GGCAGTGGCACAGGCGGTGG
20
1615
|
|
1131148
696
715
4925
4944
CCCGCGGCCATCATAGCAGC
57
1616
|
|
1131176
749
768
4978
4997
GGCTGACAGGGCGCACCCGA
31
1617
|
|
1131204
802
821
5031
5050
AGTTCCGCGCTTGCTCGGCA
13
1618
|
|
1131232
886
905
5200
5219
TCAGCCGGTCGCGGTTCAGC
39
1619
|
|
1131260
974
993
5288
5307
AGTGGGACATGAAGCCTAGG
34
1620
|
|
1131288
1060
1079
N/A
N/A
TCGCCGGCAAGGCTCCCGGG
68
1621
|
|
1131316
1095
1114
5514
5533
GCCGTTCCTGGTCAGGGAAG
27
1622
|
|
1131344
1138
1157
5557
5576
TCGAAGACAGACTCTTGCGG
15
1623
|
|
1131372
1212
1231
5631
5650
GTGGCCCCAGTACAGCGCGG
32
1624
|
|
1131400
1285
1304
N/A
N/A
CGGGCCGGTCCTGCAGGCAG
34
1625
|
|
1131428
1351
1370
6011
6030
TCTGGCACGGCTCACAGCTG
17
1626
|
|
1131456
1441
1460
6184
6203
CATCCTCCTGAAGGCGCAAC
51
1627
|
|
1131484
1500
1519
6243
6262
GCCGCTTGGCAGGCACACCG
17
1628
|
|
1131512
1567
1586
N/A
N/A
CCGCCCCCTCGAACTGGTGG
125
1629
|
|
1131540
1609
1628
6907
6926
GGAACGGTACCTGCGCCTCC
62
1630
|
|
1131568
1701
1720
6999
7018
ATCGGTGCCGCCCTCGAGGA
43†
1631
|
|
1131596
1783
1802
7171
7190
TGATGATGCCTTGCAGGGTG
21
1632
|
|
1131624
1812
1831
7200
7219
GTTGCGGTCACCACAGCCCG
20
1633
|
|
1131652
1860
1879
7248
7267
CCGGATCCAGGCCAGGTAGT
30
1634
|
|
1131680
1898
1917
7286
7305
AAAGATGAGTCCCTGAGCAA
20
1635
|
|
1131708
1940
1959
7328
7347
CATGCCCCAGCCACTCTCTC
25
1636
|
|
1131736
1993
2012
7381
7400
CATCCTGGCGCGGAGCTGGC
16
1637
|
|
1131764
2027
2046
7415
7434
AGCATTTTCAAAGCACTTTA
9
97
|
|
1131792
N/A
N/A
148
167
TGGCTGTGATAGCGACCCCC
50
1638
|
|
1131820
N/A
N/A
244
263
GTCTCCTCTGCCTGGGCCTT
61
1639
|
|
1131848
N/A
N/A
335
354
AACCCTTTCTACCTCCCCCC
76
1640
|
|
1131876
N/A
N/A
425
444
GACTATAAGTCATAGCCTGC
145
1641
|
|
1131904
N/A
N/A
569
588
CTGCACCATACACATCCCCA
37
1642
|
|
1131932
N/A
N/A
660
679
GATCACACAGCTCACGATCA
54
1643
|
|
1131960
N/A
N/A
779
798
GGAAACTGAGGAAGGCTGGC
85
1644
|
|
1131988
N/A
N/A
922
941
TGATAAAGCTGGGCTTCAGA
69
1645
|
|
1132016
N/A
N/A
1031
1050
CCTTCTCAACAACCCTAAGA
59
1646
|
|
1132044
N/A
N/A
1129
1148
GGTACCCTTCATCTAAAAGG
56
1647
|
|
1132072
N/A
N/A
1545
1564
TCCATCTGGTTCACAGGCCC
66
1648
|
|
1132100
N/A
N/A
1676
1695
CAAGAGAGAGGAGAAACTTT
108
1649
|
|
1132128
N/A
N/A
2090
2109
ATGGATCTGAGATGGACCTT
34
1650
|
|
1132156
N/A
N/A
2194
2213
AGTGTCTGGCACAAAGTACC
28
1651
|
|
1132184
N/A
N/A
2311
2330
TTCAATGTTCACTGTGATTT
52
1652
|
|
1132212
N/A
N/A
2446
2465
GCATTGATGAATCAGCAACA
55
1653
|
|
1132240
N/A
N/A
2560
2579
CTCTGAGCCTCCTGCTGAAG
33
1654
|
|
1132268
N/A
N/A
2697
2716
AGTTAAGGTTCAACAAGGCG
36
1655
|
|
1132296
N/A
N/A
2802
2821
GTGCCTACTTGCTGAATTTT
68
1656
|
|
1132324
N/A
N/A
2908
2927
AGTCACTGTTAACCACTCTG
41
1657
|
|
1132352
N/A
N/A
3012
3031
TCCACAACCTGCTAGCTGTG
29
1658
|
|
1132380
N/A
N/A
3406
3425
CTCAAAAAGGTCGCTGTGCA
36
1659
|
|
1132408
N/A
N/A
3493
3512
CAGGGACAACACACTCTCTG
86
1660
|
|
1132436
N/A
N/A
3691
3710
CAGTCCCCTCTCTCCAAGCA
55
1661
|
|
1132464
N/A
N/A
3859
3878
AGAGCCCCAGGCCACCCCAG
39
1662
|
|
1132492
N/A
N/A
3981
4000
TCCTTCCCAGAACTCTCCCT
53
1663
|
|
1132520
N/A
N/A
4069
4088
GCCAAGGCCCTTCTTGAACC
25
1664
|
|
1132548
N/A
N/A
4269
4288
CAGCACCCCGCCCAGGTCCT
73
1665
|
|
1132576
N/A
N/A
4505
4524
CCCTGGCTGGCCAGCCGCTG
46
1666
|
|
1132604
N/A
N/A
4602
4621
CCCCCCCAGAGAGCTCTCCT
120
1667
|
|
1132632
N/A
N/A
4819
4838
TCCCTCCTCCTTCCTGGCAA
30
1668
|
|
1132660
N/A
N/A
5112
5131
CCGGGAGCCCGGAGCCCTGG
68
1669
|
|
1132688
N/A
N/A
5471
5490
AAGGCTGTGGAGGAGCAGGG
69
1670
|
|
1132716
N/A
N/A
5877
5896
CCCTGCTCCCGAACCCCAAT
69
1671
|
|
1132744
N/A
N/A
6343
6362
GTCTCCTCCCCTACCCCTGC
63
1672
|
|
1132772
N/A
N/A
6445
6464
CTACACATTCTCACAACCCA
57
1673
|
|
1132800
N/A
N/A
6569
6588
AGTTGGCCTTTGCAGCCCGG
34
1674
|
|
1132828
N/A
N/A
6676
6695
CCAGGATTTGAATGGGCGGA
65
1675
|
|
1132856
N/A
N/A
6782
6801
CGCTTGTAAACCCACTCATG
51
1676
|
|
1132884
N/A
N/A
7063
7082
TGAGATTTGTGCCTGACGGC
126†
1677
|
|
TABLE 26
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed
|
PO/PS internucleoside linkages (Huh7, electroporation,
|
5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130757
21
40
21
40
CCGTTGGTCCAGCTGCCTAT
42
1678
|
|
1130785
69
88
69
88
GCTCACCAGCAGGAACCCCA
25
1679
|
|
1130813
127
146
494
513
CTTTGTACTTATGCTCCTTG
55
1680
|
|
1130841
161
180
N/A
N/A
CCGGTGACAGTGAGAACGAC
11
1681
|
|
1130869
214
233
3565
3584
TACATTTGTGGTACAGCTGC
13
1682
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
10
183
|
|
1130898
281
300
3788
3807
CGCTGGTCCTGATCAAAGTT
14
1683
|
|
1130926
320
339
N/A
N/A
TGGTCTTTCACTTTCTTGGG
23
1684
|
|
1130954
377
396
4185
4204
CCGCTTGGCATGTTCACACA
15
1685
|
|
1130982
435
454
N/A
N/A
GCACTTCTCTTTCTGGCAGT
21
1686
|
|
1131010
476
495
4421
4440
ATCTCATTCTTGTGGAAAAA
29
1687
|
|
1131038
517
536
4462
4481
ACTGGCATCTGGCCACAGCT
36
1688
|
|
1131066
556
575
4501
4520
GGCTGGCCAGCCGCTGGCAG
51
1689
|
|
1131094
594
613
4678
4697
GCGACCCCCATGGAGGCACG
26
1690
|
|
1131122
635
654
4719
4738
ACCGGGCAGTGGCACAGGCG
50
1691
|
|
1131150
698
717
4927
4946
AGCCCGCGGCCATCATAGCA
56
1692
|
|
1131178
751
770
4980
4999
ACGGCTGACAGGGCGCACCC
29
52
|
|
1131206
805
824
5034
5053
CCCAGTTCCGCGCTTGCTCG
28
1693
|
|
1131234
889
908
5203
5222
AGCTCAGCCGGTCGCGGTTC
25
1694
|
|
1131262
976
995
5290
5309
TGAGTGGGACATGAAGCCTA
25
1695
|
|
1131290
1062
1081
N/A
N/A
CTTCGCCGGCAAGGCTCCCG
36
1696
|
|
1131318
1104
1123
5523
5542
GCTCAGTGGGCCGTTCCTGG
31
1697
|
|
1131346
1140
1159
5559
5578
CATCGAAGACAGACTCTTGC
15
1698
|
|
1131374
1215
1234
5634
5653
ACTGTGGCCCCAGTACAGCG
22
1699
|
|
1131402
1288
1307
N/A
N/A
GTGCGGGCCGGTCCTGCAGG
79
1700
|
|
1131430
1353
1372
6013
6032
CGTCTGGCACGGCTCACAGC
22
1701
|
|
1131458
1443
1462
6186
6205
CGCATCCTCCTGAAGGCGCA
11
1702
|
|
1131486
1520
1539
6263
6282
GTCTCGGAGGGTCGCGCGGC
47
1703
|
|
1131514
1569
1588
N/A
N/A
CTCCGCCCCCTCGAACTGGT
61
1704
|
|
1131542
1612
1631
6910
6929
AGAGGAACGGTACCTGCGCC
155
1705
|
|
1131570
1704
1723
7002
7021
CGCATCGGTGCCGCCCTCGA
20†
1706
|
|
1131598
1785
1804
7173
7192
GCTGATGATGCCTTGCAGGG
21
1707
|
|
1131626
1815
1834
7203
7222
CTTGTTGCGGTCACCACAGC
53
1708
|
|
1131654
1862
1881
7250
7269
TCCCGGATCCAGGCCAGGTA
60
1709
|
|
1131682
1905
1924
7293
7312
AGGAGGGAAAGATGAGTCCC
17
1710
|
|
1131710
1942
1961
7330
7349
TCCATGCCCCAGCCACTCTC
10
1711
|
|
1131738
1995
2014
7383
7402
GCCATCCTGGCGCGGAGCTG
21
157
|
|
1131766
2029
2048
7417
7436
TCAGCATTTTCAAAGCACTT
9
1712
|
|
1131794
N/A
N/A
154
173
GGCTCATGGCTGTGATAGCG
36
1713
|
|
1131822
N/A
N/A
250
269
TTGTCTGTCTCCTCTGCCTG
79
1714
|
|
1131850
N/A
N/A
341
360
GACCCAAACCCTTTCTACCT
37
1715
|
|
1131878
N/A
N/A
431
450
GGAACTGACTATAAGTCATA
68
1716
|
|
1131906
N/A
N/A
576
595
GCACACACTGCACCATACAC
74
1717
|
|
1131934
N/A
N/A
666
685
CTCAAGGATCACACAGCTCA
55
1718
|
|
1131962
N/A
N/A
835
854
GCATGAATGATGCCCATGAG
85
1719
|
|
1131990
N/A
N/A
928
947
CATAAGTGATAAAGCTGGGC
42
1720
|
|
1132018
N/A
N/A
1037
1056
GGTTTCCCTTCTCAACAACC
58
1721
|
|
1132046
N/A
N/A
1135
1154
ATGGCAGGTACCCTTCATCT
85
1722
|
|
1132074
N/A
N/A
1551
1570
GTAAGGTCCATCTGGTTCAC
61
1723
|
|
1132102
N/A
N/A
1682
1701
GTAAGGCAAGAGAGAGGAGA
35
1724
|
|
1132130
N/A
N/A
2096
2115
ATGGAAATGGATCTGAGATG
53
1725
|
|
1132158
N/A
N/A
2204
2223
AATGCTGAGCAGTGTCTGGC
59
1726
|
|
1132186
N/A
N/A
2317
2336
TGCTAGTTCAATGTTCACTG
31
1727
|
|
1132214
N/A
N/A
2452
2471
AGCTTAGCATTGATGAATCA
49
1728
|
|
1132242
N/A
N/A
2567
2586
AAATGGCCTCTGAGCCTCCT
67
1729
|
|
1132270
N/A
N/A
2703
2722
ATTCCCAGTTAAGGTTCAAC
45
1730
|
|
1132298
N/A
N/A
2808
2827
GAGAATGTGCCTACTTGCTG
39
1731
|
|
1132326
N/A
N/A
2914
2933
TGTTGAAGTCACTGTTAACC
91
1732
|
|
1132354
N/A
N/A
3018
3037
CCTGACTCCACAACCTGCTA
36
1733
|
|
1132382
N/A
N/A
3412
3431
CAGACCCTCAAAAAGGTCGC
29
1734
|
|
1132410
N/A
N/A
3502
3521
TGAGAACTGCAGGGACAACA
76
1735
|
|
1132438
N/A
N/A
3697
3716
CTATCACAGTCCCCTCTCTC
64
1736
|
|
1132466
N/A
N/A
3865
3884
GAGGAGAGAGCCCCAGGCCA
51
1737
|
|
1132494
N/A
N/A
3987
4006
GACCACTCCTTCCCAGAACT
56
1738
|
|
1132522
N/A
N/A
4093
4112
GGGAGCTGAGTCACACAGCT
67
1739
|
|
1132550
N/A
N/A
4283
4302
TTGCCCCTGTCCCCCAGCAC
71
1740
|
|
1132578
N/A
N/A
4511
4530
TGCTCACCCTGGCTGGCCAG
48
1741
|
|
1132606
N/A
N/A
4608
4627
TAAAGACCCCCCCAGAGAGC
30
1742
|
|
1132634
N/A
N/A
4826
4845
CGCACTCTCCCTCCTCCTTC
51
1743
|
|
1132662
N/A
N/A
5146
5165
TCCTGTAGCCACACGACGGG
52
1744
|
|
1132690
N/A
N/A
5480
5499
TTCGCCGGCAAGGCTGTGGA
64
1745
|
|
1132718
N/A
N/A
5883
5902
GAAGCCCCCTGCTCCCGAAC
26
1746
|
|
1132746
N/A
N/A
6349
6368
TCAAAGGTCTCCTCCCCTAC
45
1747
|
|
1132774
N/A
N/A
6451
6470
ATTCACCTACACATTCTCAC
82
1748
|
|
1132802
N/A
N/A
6575
6594
CGTCCTAGTTGGCCTTTGCA
86
1749
|
|
1132830
N/A
N/A
6685
6704
AGAGAAAAGCCAGGATTTGA
93
1750
|
|
1132858
N/A
N/A
6802
6821
ATGGACAAAGCTGCTCCAGG
30
1751
|
|
1132886
N/A
N/A
7069
7088
TGGACCTGAGATTTGTGCCT
77†
1752
|
|
TABLE 27
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed
|
PO/PS internucleoside linkages (Huh7, electroporation
|
5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5' to 3')
(% UTC)
NO
|
|
1130784
68
87
68
87
CTCACCAGCAGGAACCCCAG
106
1753
|
|
1130812
126
145
493
512
TTTGTACTTATGCTCCTTGG
18
1754
|
|
1130840
160
179
N/A
N/A
CGGTGACAGTGAGAACGACT
18
1755
|
|
1130868
213
232
3564
3583
ACATTTGTGGTACAGCTGCC
18
1756
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
16
183
|
|
1130897
280
299
3787
3806
GCTGGTCCTGATCAAAGTTG
22
1757
|
|
1130925
319
338
N/A
N/A
GGTCTTTCACTTTCTTGGGC
16
1758
|
|
1130953
376
395
4184
4203
CGCTTGGCATGTTCACACAG
40
1759
|
|
1130981
434
453
N/A
N/A
CACTTCTCTTTCTGGCAGTG
15
1760
|
|
1131009
475
494
4420
4439
TCTCATTCTTGTGGAAAAAC
18
1761
|
|
1131037
516
535
4461
4480
CTGGCATCTGGCCACAGCTG
25
1762
|
|
1131065
555
574
4500
4519
GCTGGCCAGCCGCTGGCAGT
131
1763
|
|
1131093
592
611
4676
4695
GACCCCCATGGAGGCACGGG
36
1764
|
|
1131121
633
652
4717
4736
CGGGCAGTGGCACAGGCGGT
20
1765
|
|
1131149
697
716
4926
4945
GCCCGCGGCCATCATAGCAG
35
1766
|
|
1131177
750
769
4979
4998
CGGCTGACAGGGCGCACCCG
26
1767
|
|
1131205
804
823
5033
5052
CCAGTTCCGCGCTTGCTCGG
26
1768
|
|
1131233
888
907
5202
5221
GCTCAGCCGGTCGCGGTTCA
21
1769
|
|
1131261
975
994
5289
5308
GAGTGGGACATGAAGCCTAG
24
1770
|
|
1131289
1061
1080
N/A
N/A
TTCGCCGGCAAGGCTCCCGG
40
1771
|
|
1131317
1102
1121
5521
5540
TCAGTGGGCCGTTCCTGGTC
31
1772
|
|
1131345
1139
1158
5558
5577
ATCGAAGACAGACTCTTGCG
19
1773
|
|
1131373
1213
1232
5632
5651
TGTGGCCCCAGTACAGCGCG
43
1774
|
|
1131401
1286
1305
N/A
N/A
GCGGGCCGGTCCTGCAGGCA
28
1775
|
|
1131429
1352
1371
6012
6031
GTCTGGCACGGCTCACAGCT
16
1776
|
|
1131457
1442
1461
6185
6204
GCATCCTCCTGAAGGCGCAA
24
1777
|
|
1131485
1501
1520
6244
6263
CGCCGCTTGGCAGGCACACC
45
1778
|
|
1131513
1568
1587
N/A
N/A
TCCGCCCCCTCGAACTGGTG
70
1779
|
|
1131541
1610
1629
6908
6927
AGGAACGGTACCTGCGCCTC
66
1780
|
|
1131597
1784
1803
7172
7191
CTGATGATGCCTTGCAGGGT
24
1781
|
|
1131625
1813
1832
7201
7220
TGTTGCGGTCACCACAGCCC
15
1782
|
|
1131653
1861
1880
7249
7268
CCCGGATCCAGGCCAGGTAG
40
1783
|
|
1131681
1901
1920
7289
7308
GGGAAAGATGAGTCCCTGAG
19
92
|
|
1131709
1941
1960
7329
7348
CCATGCCCCAGCCACTCTCT
18
1784
|
|
1131737
1994
2013
7382
7401
CCATCCTGGCGCGGAGCTGG
17
1785
|
|
1131765
2028
2047
7416
7435
CAGCATTTTCAAAGCACTTT
39
1786
|
|
1131793
N/A
N/A
151
170
TCATGGCTGTGATAGCGACC
80
1787
|
|
1131821
N/A
N/A
247
266
TCTGTCTCCTCTGCCTGGGC
91
1788
|
|
1131849
N/A
N/A
338
357
CCAAACCCTTTCTACCTCCC
103
1789
|
|
1131877
N/A
N/A
428
447
ACTGACTATAAGTCATAGCC
77
1790
|
|
1131905
N/A
N/A
572
591
ACACTGCACCATACACATCC
59
1791
|
|
1131933
N/A
N/A
663
682
AAGGATCACACAGCTCACGA
64
1792
|
|
1131961
N/A
N/A
832
851
TGAATGATGCCCATGAGACG
33
1793
|
|
1131989
N/A
N/A
925
944
AAGTGATAAAGCTGGGCTTC
61
1794
|
|
1132017
N/A
N/A
1034
1053
TTCCCTTCTCAACAACCCTA
49
1795
|
|
1132045
N/A
N/A
1132
1151
GCAGGTACCCTTCATCTAAA
44
1796
|
|
1132073
N/A
N/A
1548
1567
AGGTCCATCTGGTTCACAGG
41
1797
|
|
1132101
N/A
N/A
1679
1698
AGGCAAGAGAGAGGAGAAAC
37
1798
|
|
1132129
N/A
N/A
2093
2112
GAAATGGATCTGAGATGGAC
65
1799
|
|
1132157
N/A
N/A
2201
2220
GCTGAGCAGTGTCTGGCACA
35
1800
|
|
1132185
N/A
N/A
2314
2333
TAGTTCAATGTTCACTGTGA
44
1801
|
|
1132213
N/A
N/A
2449
2468
TTAGCATTGATGAATCAGCA
46
1802
|
|
1132241
N/A
N/A
2564
2583
TGGCCTCTGAGCCTCCTGCT
60
1803
|
|
1132269
N/A
N/A
2700
2719
CCCAGTTAAGGTTCAACAAG
62
1804
|
|
1132297
N/A
N/A
2805
2824
AATGTGCCTACTTGCTGAAT
77
1805
|
|
1132325
N/A
N/A
2911
2930
TGAAGTCACTGTTAACCACT
45
1806
|
|
1132353
N/A
N/A
3015
3034
GACTCCACAACCTGCTAGCT
62
1807
|
|
1132381
N/A
N/A
3409
3428
ACCCTCAAAAAGGTCGCTGT
58
1808
|
|
1132409
N/A
N/A
3499
3518
GAACTGCAGGGACAACACAC
48
1809
|
|
1132437
N/A
N/A
3694
3713
TCACAGTCCCCTCTCTCCAA
63
1810
|
|
1132465
N/A
N/A
3862
3881
GAGAGAGCCCCAGGCCACCC
41
1811
|
|
1132493
N/A
N/A
3984
4003
CACTCCTTCCCAGAACTCTC
52
1812
|
|
1132521
N/A
N/A
4090
4109
AGCTGAGTCACACAGCTGGG
69
1813
|
|
1132549
N/A
N/A
4272
4291
CCCCAGCACCCCGCCCAGGT
113
1814
|
|
1132577
N/A
N/A
4508
4527
TCACCCTGGCTGGCCAGCCG
103
1815
|
|
1132605
N/A
N/A
4605
4624
AGACCCCCCCAGAGAGCTCT
72
1816
|
|
1132633
N/A
N/A
4823
4842
ACTCTCCCTCCTCCTTCCTG
42
1817
|
|
1132661
N/A
N/A
5115
5134
GCGCCGGGAGCCCGGAGCCC
92
1818
|
|
1132689
N/A
N/A
5474
5493
GGCAAGGCTGTGGAGGAGCA
82
1819
|
|
1132717
N/A
N/A
5880
5899
GCCCCCTGCTCCCGAACCCC
65
1820
|
|
1132745
N/A
N/A
6346
6365
AAGGTCTCCTCCCCTACCCC
68
1821
|
|
1132773
N/A
N/A
6448
6467
CACCTACACATTCTCACAAC
66
1822
|
|
1132801
N/A
N/A
6572
6591
CCTAGTTGGCCTTTGCAGCC
67
1823
|
|
1132829
N/A
N/A
6679
6698
AAGCCAGGATTTGAATGGGC
50
1824
|
|
1132857
N/A
N/A
6799
6818
GACAAAGCTGCTCCAGGCGC
74
1825
|
|
1131569
1703
1722
7001
7020
GCATCGGTGCCGCCCTCGAG
8†
1826
|
|
1132885
N/A
N/A
7066
7085
ACCTGAGATTTGTGCCTGAC
72†
1827
|
|
TABLE 28
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130758
22
41
22
41
TCCGTTGGTCCAGCTGCCTA
70
1828
|
|
1130786
70
89
70
89
AGCTCACCAGCAGGAACCCC
55
1829
|
|
1130814
128
147
495
514
GCTTTGTACTTATGCTCCTT
30
1830
|
|
1130842
162
181
N/A
N/A
CCCGGTGACAGTGAGAACGA
43
1831
|
|
1130870
215
234
3566
3585
GTACATTTGTGGTACAGCTG
19
182
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
10
183
|
|
1130899
282
301
3789
3808
TCGCTGGTCCTGATCAAAGT
30
1832
|
|
1130927
322
341
N/A
N/A
AGTGGTCTTTCACTTTCTTG
41
1833
|
|
1130955
378
397
4186
4205
GCCGCTTGGCATGTTCACAC
46
1834
|
|
1130983
436
455
N/A
N/A
AGCACTTCTCTTTCTGGCAG
14
1835
|
|
1131011
478
497
4423
4442
ATATCTCATTCTTGTGGAAA
39
1836
|
|
1131039
524
543
4469
4488
CCCTTGCACTGGCATCTGGC
38
1837
|
|
1131067
557
576
4502
4521
TGGCTGGCCAGCCGCTGGCA
45
1838
|
|
1131095
595
614
4679
4698
AGCGACCCCCATGGAGGCAC
21
1839
|
|
1131123
637
656
4721
4740
CCACCGGGCAGTGGCACAGG
42
1840
|
|
1131151
699
718
4928
4947
GAGCCCGCGGCCATCATAGC
85
1841
|
|
1131179
752
771
4981
5000
CACGGCTGACAGGGCGCACC
94
1842
|
|
1131207
808
827
5037
5056
GTCCCCAGTTCCGCGCTTGC
90
53
|
|
1131235
891
910
5205
5224
CCAGCTCAGCCGGTCGCGGT
72
1843
|
|
1131263
977
996
5291
5310
ATGAGTGGGACATGAAGCCT
51
1844
|
|
1131291
1063
1082
N/A
N/A
GCTTCGCCGGCAAGGCTCCC
41
1845
|
|
1131319
1106
1125
5525
5544
CAGCTCAGTGGGCCGTTCCT
66
1846
|
|
1131347
1141
1160
5560
5579
TCATCGAAGACAGACTCTTG
27
1847
|
|
1131375
1216
1235
5635
5654
AACTGTGGCCCCAGTACAGC
17
1848
|
|
1131403
1289
1308
N/A
N/A
GGTGCGGGCCGGTCCTGCAG
79
1849
|
|
1131431
1371
1390
6031
6050
GTAGGAGCGCACGGCCAACG
46
1850
|
|
1131459
1445
1464
6188
6207
TCCGCATCCTCCTGAAGGCG
40
1851
|
|
1131487
1521
1540
6264
6283
GGTCTCGGAGGGTCGCGCGG
61
1852
|
|
1131515
1570
1589
N/A
N/A
CCTCCGCCCCCTCGAACTGG
258
1853
|
|
1131543
1613
1632
6911
6930
GAGAGGAACGGTACCTGCGC
80
1854
|
|
1131571
1707
1726
7005
7024
GCACGCATCGGTGCCGCCCT
22†
1855
|
|
1131599
1786
1805
7174
7193
AGCTGATGATGCCTTGCAGG
25
1856
|
|
1131627
1816
1835
7204
7223
GCTTGTTGCGGTCACCACAG
21
1857
|
|
1131655
1863
1882
7251
7270
CTCCCGGATCCAGGCCAGGT
81
1858
|
|
1131683
1906
1925
7294
7313
AAGGAGGGAAAGATGAGTCC
48
1859
|
|
1131711
1943
1962
7331
7350
TTCCATGCCCCAGCCACTCT
20
1860
|
|
1131739
1996
2015
7384
7403
CGCCATCCTGGCGCGGAGCT
15
1861
|
|
1131795
N/A
N/A
157
176
CATGGCTCATGGCTGTGATA
82
1862
|
|
1131823
N/A
N/A
253
272
TGGTTGTCTGTCTCCTCTGC
70
1863
|
|
1131851
N/A
N/A
344
363
CAAGACCCAAACCCTTTCTA
154
1864
|
|
1131879
N/A
N/A
434
453
CAGGGAACTGACTATAAGTC
74
1865
|
|
1131907
N/A
N/A
579
598
ACTGCACACACTGCACCATA
78
1866
|
|
1131935
N/A
N/A
669
688
GCCCTCAAGGATCACACAGC
80
1867
|
|
1131963
N/A
N/A
838
857
GTGGCATGAATGATGCCCAT
153
1868
|
|
1131991
N/A
N/A
931
950
GCTCATAAGTGATAAAGCTG
36
1869
|
|
1132019
N/A
N/A
1043
1062
CACATGGGTTTCCCTTCTCA
100
1870
|
|
1132047
N/A
N/A
1138
1157
GAAATGGCAGGTACCCTTCA
46
1871
|
|
1132075
N/A
N/A
1554
1573
AGGGTAAGGTCCATCTGGTT
85
1872
|
|
1132103
N/A
N/A
1688
1707
GTTTGGGTAAGGCAAGAGAG
47
1873
|
|
1132131
N/A
N/A
2099
2118
TAAATGGAAATGGATCTGAG
37
1874
|
|
1132159
N/A
N/A
2207
2226
TGAAATGCTGAGCAGTGTCT
84
1875
|
|
1132187
N/A
N/A
2320
2339
GTTTGCTAGTTCAATGTTCA
56
1876
|
|
1132215
N/A
N/A
2455
2474
ATCAGCTTAGCATTGATGAA
44
1877
|
|
1132243
N/A
N/A
2570
2589
TGGAAATGGCCTCTGAGCCT
52
1878
|
|
1132271
N/A
N/A
2706
2725
CACATTCCCAGTTAAGGTTC
30
1879
|
|
1132299
N/A
N/A
2811
2830
ATTGAGAATGTGCCTACTTG
87
1880
|
|
1132327
N/A
N/A
2917
2936
GAATGTTGAAGTCACTGTTA
74
1881
|
|
1132355
N/A
N/A
3021
3040
ACTCCTGACTCCACAACCTG
49
1882
|
|
1132383
N/A
N/A
3415
3434
GGACAGACCCTCAAAAAGGT
72
1883
|
|
1132411
N/A
N/A
3505
3524
CAGTGAGAACTGCAGGGACA
73
1884
|
|
1132439
N/A
N/A
3700
3719
GCCCTATCACAGTCCCCTCT
61
1885
|
|
1132467
N/A
N/A
3868
3887
CGGGAGGAGAGAGCCCCAGG
46
1886
|
|
1132495
N/A
N/A
3990
4009
TGGGACCACTCCTTCCCAGA
65
1887
|
|
1132523
N/A
N/A
4096
4115
GCAGGGAGCTGAGTCACACA
121
1888
|
|
1132551
N/A
N/A
4286
4305
GGGTTGCCCCTGTCCCCCAG
59
1889
|
|
1132579
N/A
N/A
4514
4533
ATCTGCTCACCCTGGCTGGC
48
1890
|
|
1132607
N/A
N/A
4611
4630
GCCTAAAGACCCCCCCAGAG
75
1891
|
|
1132635
N/A
N/A
4832
4851
GCTTTCCGCACTCTCCCTCC
54
1892
|
|
1132663
N/A
N/A
5149
5168
GGTTCCTGTAGCCACACGAC
86
1893
|
|
1132691
N/A
N/A
5483
5502
CGCTTCGCCGGCAAGGCTGT
41
1894
|
|
1132719
N/A
N/A
5910
5929
CTCAGACCTGGCCACAAGCG
89
1895
|
|
1132747
N/A
N/A
6352
6371
TGATCAAAGGTCTCCTCCCC
118
1896
|
|
1132775
N/A
N/A
6454
6473
GGGATTCACCTACACATTCT
119
1897
|
|
1132803
N/A
N/A
6600
6619
ACACTAGCCCGGAGCGCGGG
62
1898
|
|
1132831
N/A
N/A
6688
6707
TCCAGAGAAAAGCCAGGATT
86
1899
|
|
1132859
N/A
N/A
6805
6824
ACGATGGACAAAGCTGCTCC
55
1900
|
|
1132887
N/A
N/A
7072
7091
CTGTGGACCTGAGATTTGTG
155†
1901
|
|
TABLE 29
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130759
24
43
24
43
CGTCCGTTGGTCCAGCTGCC
35
1902
|
|
1130787
71
90
71
90
AAGCTCACCAGCAGGAACCC
47
1903
|
|
1130815
129
148
496
515
AGCTTTGTACTTATGCTCCT
44
1904
|
|
1130843
163
182
N/A
N/A
CCCCGGTGACAGTGAGAACG
27
1905
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
17
183
|
|
1130872
217
236
3568
3587
GGGTACATTTGTGGTACAGC
23
25
|
|
1130900
283
302
3790
3809
ATCGCTGGTCCTGATCAAAG
22
1906
|
|
1130928
323
342
N/A
N/A
CAGTGGTCTTTCACTTTCTT
12
1907
|
|
1130956
379
398
4187
4206
GGCCGCTTGGCATGTTCACA
66
1908
|
|
1130984
438
457
N/A
N/A
AAAGCACTTCTCTTTCTGGC
33
1909
|
|
1131012
479
498
4424
4443
CATATCTCATTCTTGTGGAA
38
1910
|
|
1131040
525
544
4470
4489
ACCCTTGCACTGGCATCTGG
22
1911
|
|
1131068
558
577
4503
4522
CTGGCTGGCCAGCCGCTGGC
26
1912
|
|
1131096
597
616
4681
4700
GCAGCGACCCCCATGGAGGC
23
1913
|
|
1131124
638
657
4722
4741
CCCACCGGGCAGTGGCACAG
45
1914
|
|
1131152
701
720
4930
4949
CTGAGCCCGCGGCCATCATA
35
1915
|
|
1131180
753
772
4982
5001
CCACGGCTGACAGGGCGCAC
36
1916
|
|
1131208
812
831
5041
5060
CCCAGTCCCCAGTTCCGCGC
22
1917
|
|
1131236
892
911
5206
5225
CCCAGCTCAGCCGGTCGCGG
71
1918
|
|
1131264
979
998
5293
5312
GCATGAGTGGGACATGAAGC
47
1919
|
|
1131292
1064
1083
N/A
N/A
CGCTTCGCCGGCAAGGCTCC
55
1920
|
|
1131320
1107
1126
5526
5545
GCAGCTCAGTGGGCCGTTCC
71
1921
|
|
1131348
1143
1162
5562
5581
GGTCATCGAAGACAGACTCT
22
1922
|
|
1131376
1218
1237
5637
5656
GAAACTGTGGCCCCAGTACA
32
1923
|
|
1131404
1302
1321
5962
5981
CGTCAGATCCTCGGGTGCGG
29
1924
|
|
1131432
1372
1391
6032
6051
GGTAGGAGCGCACGGCCAAC
22
1925
|
|
1131460
1446
1465
6189
6208
GTCCGCATCCTCCTGAAGGC
31
1926
|
|
1131488
1522
1541
6265
6284
TGGTCTCGGAGGGTCGCGCG
40
1927
|
|
1131516
1571
1590
N/A
N/A
TCCTCCGCCCCCTCGAACTG
425
1928
|
|
1131544
1614
1633
6912
6931
GGAGAGGAACGGTACCTGCG
87
1929
|
|
1131572
1709
1728
7007
7026
TGGCACGCATCGGTGCCGCC
63†
1930
|
|
1131600
1787
1806
7175
7194
CAGCTGATGATGCCTTGCAG
90
1931
|
|
1131628
1817
1836
7205
7224
GGCTTGTTGCGGTCACCACA
40
1932
|
|
1131656
1864
1883
7252
7271
GCTCCCGGATCCAGGCCAGG
56
1933
|
|
1131684
1907
1926
7295
7314
CAAGGAGGGAAAGATGAGTC
33
1934
|
|
1131712
1944
1963
7332
7351
CTTCCATGCCCCAGCCACTC
20
1935
|
|
1131740
1997
2016
7385
7404
GCGCCATCCTGGCGCGGAGC
38
1936
|
|
1131796
N/A
N/A
160
179
GGCCATGGCTCATGGCTGTG
79
1937
|
|
1131824
N/A
N/A
256
275
GTCTGGTTGTCTGTCTCCTC
63
1938
|
|
1131852
N/A
N/A
347
366
TCCCAAGACCCAAACCCTTT
57
1939
|
|
1131880
N/A
N/A
437
456
AGGCAGGGAACTGACTATAA
33
1940
|
|
1131908
N/A
N/A
582
601
GAGACTGCACACACTGCACC
40
1941
|
|
1131936
N/A
N/A
672
691
CCTGCCCTCAAGGATCACAC
45
1942
|
|
1131964
N/A
N/A
841
860
TGTGTGGCATGAATGATGCC
85
1943
|
|
1131992
N/A
N/A
934
953
CCAGCTCATAAGTGATAAAG
36
1944
|
|
1132020
N/A
N/A
1046
1065
CCTCACATGGGTTTCCCTTC
26
1945
|
|
1132048
N/A
N/A
1159
1178
TGGCAGAGCTTGAGGAAGGG
31
1946
|
|
1132076
N/A
N/A
1557
1576
ACCAGGGTAAGGTCCATCTG
65
1947
|
|
1132104
N/A
N/A
1691
1710
CAAGTTTGGGTAAGGCAAGA
40
1948
|
|
1132132
N/A
N/A
2102
2121
CAGTAAATGGAAATGGATCT
64
1949
|
|
1132160
N/A
N/A
2210
2229
CCATGAAATGCTGAGCAGTG
46
1950
|
|
1132188
N/A
N/A
2323
2342
ACTGTTTGCTAGTTCAATGT
25
1951
|
|
1132216
N/A
N/A
2458
2477
GCCATCAGCTTAGCATTGAT
38
1952
|
|
1132244
N/A
N/A
2573
2592
TTTTGGAAATGGCCTCTGAG
71
1953
|
|
1132272
N/A
N/A
2709
2728
TTGCACATTCCCAGTTAAGG
58
1954
|
|
1132300
N/A
N/A
2814
2833
TACATTGAGAATGTGCCTAC
116
1955
|
|
1132328
N/A
N/A
2920
2939
GTTGAATGTTGAAGTCACTG
47
1956
|
|
1132356
N/A
N/A
3024
3043
TAGACTCCTGACTCCACAAC
110
1957
|
|
1132384
N/A
N/A
3418
3437
AAAGGACAGACCCTCAAAAA
113
1958
|
|
1132412
N/A
N/A
3508
3527
TGACAGTGAGAACTGCAGGG
30
1959
|
|
1132440
N/A
N/A
3703
3722
CCTGCCCTATCACAGTCCCC
84
1960
|
|
1132468
N/A
N/A
3871
3890
AGGCGGGAGGAGAGAGCCCC
62
1961
|
|
1132496
N/A
N/A
3993
4012
AAATGGGACCACTCCTTCCC
41
1962
|
|
1132524
N/A
N/A
4099
4118
AGAGCAGGGAGCTGAGTCAC
60
1963
|
|
1132552
N/A
N/A
4289
4308
CCAGGGTTGCCCCTGTCCCC
190
1964
|
|
1132580
N/A
N/A
4517
4536
ACCATCTGCTCACCCTGGCT
67
1965
|
|
1132608
N/A
N/A
4614
4633
TGGGCCTAAAGACCCCCCCA
74
1966
|
|
1132636
N/A
N/A
4835
4854
TCTGCTTTCCGCACTCTCCC
36
1967
|
|
1132664
N/A
N/A
5152
5171
CCGGGTTCCTGTAGCCACAC
53
1968
|
|
1132692
N/A
N/A
5702
5721
CGCCGGTCCTGCAGGCAGTG
54
1969
|
|
1132720
N/A
N/A
5913
5932
GCTCTCAGACCTGGCCACAA
94
1970
|
|
1132748
N/A
N/A
6355
6374
CAGTGATCAAAGGTCTCCTC
73
1971
|
|
1132776
N/A
N/A
6457
6476
CCTGGGATTCACCTACACAT
46
1972
|
|
1132804
N/A
N/A
6603
6622
CCCACACTAGCCCGGAGCGC
52
1973
|
|
1132832
N/A
N/A
6691
6710
AGGTCCAGAGAAAAGCCAGG
60
1974
|
|
1132860
N/A
N/A
6808
6827
CGGACGATGGACAAAGCTGC
37
1975
|
|
1132888
N/A
N/A
7075
7094
GCGCTGTGGACCTGAGATTT
87†
1976
|
|
TABLE 30
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130788
72
91
72
91
CAAGCTCACCAGCAGGAACC
44
1977
|
|
1130816
130
149
497
516
CAGCTTTGTACTTATGCTCC
46
1978
|
|
1130844
164
183
3515
3534
TCCCCGGTGACAGTGAGAAC
43
24
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
12
183
|
|
1130873
218
237
3569
3588
TGGGTACATTTGTGGTACAG
22
1979
|
|
1130901
284
303
3791
3810
CATCGCTGGTCCTGATCAAA
26
1980
|
|
1130929
324
343
N/A
N/A
GCAGTGGTCTTTCACTTTCT
35
1981
|
|
1130957
380
399
4188
4207
GGGCCGCTTGGCATGTTCAC
32
1982
|
|
1130985
439
458
N/A
N/A
CAAAGCACTTCTCTTTCTGG
34
1983
|
|
1131013
481
500
4426
4445
ACCATATCTCATTCTTGTGG
21
1984
|
|
1131041
526
545
4471
4490
GACCCTTGCACTGGCATCTG
27
1985
|
|
1131069
559
578
4504
4523
CCTGGCTGGCCAGCCGCTGG
76
1986
|
|
1131097
598
617
4682
4701
GGCAGCGACCCCCATGGAGG
24
1987
|
|
1131125
640
659
4724
4743
AGCCCACCGGGCAGTGGCAC
57
1988
|
|
1131153
702
721
4931
4950
GCTGAGCCCGCGGCCATCAT
36
1989
|
|
1131181
754
773
4983
5002
CCCACGGCTGACAGGGCGCA
37
1990
|
|
1131209
813
832
5042
5061
GCCCAGTCCCCAGTTCCGCG
41
1991
|
|
1131237
893
912
5207
5226
TCCCAGCTCAGCCGGTCGCG
40
1992
|
|
1131265
980
999
5294
5313
GGCATGAGTGGGACATGAAG
28
1993
|
|
1131293
1065
1084
N/A
N/A
CCGCTTCGCCGGCAAGGCTC
35
1994
|
|
1131321
1108
1127
5527
5546
CGCAGCTCAGTGGGCCGTTC
32
1995
|
|
1131349
1144
1163
5563
5582
GGGTCATCGAAGACAGACTC
44
1996
|
|
1131377
1219
1238
5638
5657
AGAAACTGTGGCCCCAGTAC
46
1997
|
|
1131405
1303
1322
5963
5982
CCGTCAGATCCTCGGGTGCG
28
1998
|
|
1131433
1373
1392
6033
6052
CGGTAGGAGCGCACGGCCAA
21
1999
|
|
1131461
1448
1467
6191
6210
CCGTCCGCATCCTCCTGAAG
42
2000
|
|
1131489
1523
1542
6266
6285
GTGGTCTCGGAGGGTCGCGC
41
2001
|
|
1131517
1572
1591
N/A
N/A
TTCCTCCGCCCCCTCGAACT
194
2002
|
|
1131545
1615
1634
6913
6932
GGGAGAGGAACGGTACCTGC
65
2003
|
|
1131573
1710
1729
7008
7027
CTGGCACGCATCGGTGCCGC
121†
2004
|
|
1131601
1788
1807
7176
7195
CCAGCTGATGATGCCTTGCA
22
2005
|
|
1131629
1818
1837
7206
7225
TGGCTTGTTGCGGTCACCAC
26
2006
|
|
1131657
1865
1884
7253
7272
TGCTCCCGGATCCAGGCCAG
44
2007
|
|
1131685
1908
1927
7296
7315
CCAAGGAGGGAAAGATGAGT
22
2008
|
|
1131713
1945
1964
7333
7352
CCTTCCATGCCCCAGCCACT
15
2009
|
|
1131741
1999
2018
7387
7406
CTGCGCCATCCTGGCGCGGA
83
2010
|
|
1131797
N/A
N/A
163
182
AGAGGCCATGGCTCATGGCT
91
2011
|
|
1131825
N/A
N/A
259
278
CCAGTCTGGTTGTCTGTCTC
60
2012
|
|
1131853
N/A
N/A
350
369
TCTTCCCAAGACCCAAACCC
65
2013
|
|
1131881
N/A
N/A
440
459
AGAAGGCAGGGAACTGACTA
84
2014
|
|
1131909
N/A
N/A
585
604
CCTGAGACTGCACACACTGC
56
2015
|
|
1131937
N/A
N/A
675
694
TACCCTGCCCTCAAGGATCA
42
2016
|
|
1131965
N/A
N/A
844
863
CTCTGTGTGGCATGAATGAT
71
2017
|
|
1131993
N/A
N/A
937
956
CACCCAGCTCATAAGTGATA
63
2018
|
|
1132021
N/A
N/A
1049
1068
ATACCTCACATGGGTTTCCC
59
2019
|
|
1132049
N/A
N/A
1162
1181
CTATGGCAGAGCTTGAGGAA
62
2020
|
|
1132077
N/A
N/A
1560
1579
CCCACCAGGGTAAGGTCCAT
54
2021
|
|
1132105
N/A
N/A
1694
1713
CACCAAGTTTGGGTAAGGCA
34
2022
|
|
1132133
N/A
N/A
2105
2124
GGACAGTAAATGGAAATGGA
45
2023
|
|
1132161
N/A
N/A
2219
2238
GAGATAATGCCATGAAATGC
56
2024
|
|
1132189
N/A
N/A
2326
2345
AATACTGTTTGCTAGTTCAA
52
2025
|
|
1132217
N/A
N/A
2461
2480
AGTGCCATCAGCTTAGCATT
51
2026
|
|
1132245
N/A
N/A
2576
2595
GGCTTTTGGAAATGGCCTCT
46
2027
|
|
1132273
N/A
N/A
2712
2731
AATTTGCACATTCCCAGTTA
64
2028
|
|
1132301
N/A
N/A
2817
2836
TTCTACATTGAGAATGTGCC
43
2029
|
|
1132329
N/A
N/A
2923
2942
CTTGTTGAATGTTGAAGTCA
50
2030
|
|
1132357
N/A
N/A
3027
3046
GAGTAGACTCCTGACTCCAC
73
2031
|
|
1132385
N/A
N/A
3421
3440
GGAAAAGGACAGACCCTCAA
72
2032
|
|
1132413
N/A
N/A
3511
3530
CGGTGACAGTGAGAACTGCA
32
2033
|
|
1132441
N/A
N/A
3706
3725
CACCCTGCCCTATCACAGTC
74
2034
|
|
1132469
N/A
N/A
3874
3893
ATGAGGCGGGAGGAGAGAGC
72
2035
|
|
1132497
N/A
N/A
3996
4015
GCAAAATGGGACCACTCCTT
42
2036
|
|
1132525
N/A
N/A
4102
4121
GGAAGAGCAGGGAGCTGAGT
102
2037
|
|
1132553
N/A
N/A
4292
4311
GGCCCAGGGTTGCCCCTGTC
75
2038
|
|
1132581
N/A
N/A
4520
4539
CCAACCATCTGCTCACCCTG
72
2039
|
|
1132609
N/A
N/A
4617
4636
CCCTGGGCCTAAAGACCCCC
115
2040
|
|
1132637
N/A
N/A
4838
4857
TCATCTGCTTTCCGCACTCT
44
2041
|
|
1132665
N/A
N/A
5155
5174
TGTCCGGGTTCCTGTAGCCA
43
2042
|
|
1132693
N/A
N/A
5705
5724
ACTCGCCGGTCCTGCAGGCA
29
2043
|
|
1132721
N/A
N/A
6089
6108
GCCCCCACGCACCCAGGTCG
132
2044
|
|
1132749
N/A
N/A
6358
6377
ACCCAGTGATCAAAGGTCTC
56
2045
|
|
1132777
N/A
N/A
6460
6479
CTACCTGGGATTCACCTACA
51
2046
|
|
1132805
N/A
N/A
6606
6625
GCTCCCACACTAGCCCGGAG
52
2047
|
|
1132833
N/A
N/A
6694
6713
CTGAGGTCCAGAGAAAAGCC
25
2048
|
|
1132861
N/A
N/A
6811
6830
GCCCGGACGATGGACAAAGC
86
2049
|
|
1132889
N/A
N/A
7091
7110
GGAAACACGCAGCTCAGCGC
87†
2050
|
|
TABLE 31
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
No.
|
|
1130761
27
46
27
46
ATCCGTCCGTTGGTCCAGCT
24
2051
|
|
1130789
73
92
73
92
CCAAGCTCACCAGCAGGAAC
47
2052
|
|
1130817
131
150
498
517
TCAGCTTTGTACTTATGCTC
43
2053
|
|
1130845
165
184
3516
3535
CTCCCCGGTGACAGTGAGAA
68
2054
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
16
183
|
|
1130874
220
239
3571
3590
TGTGGGTACATTTGTGGTAC
21
2055
|
|
1130902
286
305
3793
3812
CCCATCGCTGGTCCTGATCA
25
2056
|
|
1130930
326
345
N/A
N/A
CTGCAGTGGTCTTTCACTTT
56
2057
|
|
1130958
400
419
4208
4227
GTTGTGGACAGAGACAGTGG
37
2058
|
|
1130986
440
459
N/A
N/A
TCAAAGCACTTCTCTTTCTG
41
2059
|
|
1131014
484
503
4429
4448
TATACCATATCTCATTCTTG
41
2060
|
|
1131042
527
546
4472
4491
GGACCCTTGCACTGGCATCT
39
2061
|
|
1131070
560
579
N/A
N/A
GCCTGGCTGGCCAGCCGCTG
82
2062
|
|
1131098
600
619
4684
4703
TAGGCAGCGACCCCCATGGA
47
2063
|
|
1131126
643
662
4727
4746
TGTAGCCCACCGGGCAGTGG
24
2064
|
|
1131154
703
722
4932
4951
AGCTGAGCCCGCGGCCATCA
29
2065
|
|
1131182
756
775
4985
5004
GGCCCACGGCTGACAGGGCG
61
2066
|
|
1131210
815
834
5044
5063
CCGCCCAGTCCCCAGTTCCG
27
2067
|
|
1131238
894
913
5208
5227
CTCCCAGCTCAGCCGGTCGC
66
2068
|
|
1131266
989
1008
5303
5322
GGCTGCGCGGGCATGAGTGG
66
2069
|
|
1131294
1066
1085
5485
5504
CCCGCTTCGCCGGCAAGGCT
49
2070
|
|
1131322
1109
1128
5528
5547
CCGCAGCTCAGTGGGCCGTT
67
2071
|
|
1131350
1146
1165
5565
5584
GCGGGTCATCGAAGACAGAC
23
2072
|
|
1131378
1220
1239
5639
5658
CAGAAACTGTGGCCCCAGTA
51
2073
|
|
1131406
1305
1324
5965
5984
CACCGTCAGATCCTCGGGTG
31
2074
|
|
1131434
1375
1394
6035
6054
AGCGGTAGGAGCGCACGGCC
24
2075
|
|
1131462
1449
1468
6192
6211
GCCGTCCGCATCCTCCTGAA
17
2076
|
|
1131490
1524
1543
6267
6286
CGTGGTCTCGGAGGGTCGCG
28
2077
|
|
1131518
1573
1592
N/A
N/A
ATTCCTCCGCCCCCTCGAAC
238
2078
|
|
1131546
1616
1635
6914
6933
AGGGAGAGGAACGGTACCTG
50
2079
|
|
1131574
1712
1731
N/A
N/A
CCCTGGCACGCATCGGTGCC
56†
2080
|
|
1131602
1789
1808
7177
7196
CCCAGCTGATGATGCCTTGC
45
2081
|
|
1131630
1819
1838
7207
7226
CTGGCTTGTTGCGGTCACCA
16
2082
|
|
1131658
1866
1885
7254
7273
GTGCTCCCGGATCCAGGCCA
32
2083
|
|
1131686
1909
1928
7297
7316
ACCAAGGAGGGAAAGATGAG
19
2084
|
|
1131714
1946
1965
7334
7353
GCCTTCCATGCCCCAGCCAC
30
2085
|
|
1131742
2000
2019
7388
7407
CCTGCGCCATCCTGGCGCGG
113
2086
|
|
1131798
N/A
N/A
166
185
AGCAGAGGCCATGGCTCATG
38
2087
|
|
1131826
N/A
N/A
262
281
CACCCAGTCTGGTTGTCTGT
71
2088
|
|
1131854
N/A
N/A
353
372
ATTTCTTCCCAAGACCCAAA
74
2089
|
|
1131882
N/A
N/A
443
462
AGAAGAAGGCAGGGAACTGA
120
2090
|
|
1131910
N/A
N/A
591
610
GACTGCCCTGAGACTGCACA
59
2091
|
|
1131938
N/A
N/A
678
697
CCATACCCTGCCCTCAAGGA
46
2092
|
|
1131966
N/A
N/A
847
866
GGCCTCTGTGTGGCATGAAT
31
2093
|
|
1131994
N/A
N/A
941
960
GAGTCACCCAGCTCATAAGT
80
2094
|
|
1132022
N/A
N/A
1052
1071
TGAATACCTCACATGGGTTT
34
2095
|
|
1132050
N/A
N/A
1165
1184
GAGCTATGGCAGAGCTTGAG
51
2096
|
|
1132078
N/A
N/A
1563
1582
AGGCCCACCAGGGTAAGGTC
61
2097
|
|
1132106
N/A
N/A
1702
1721
GGATAGGGCACCAAGTTTGG
77
2098
|
|
1132134
N/A
N/A
2108
2127
CTAGGACAGTAAATGGAAAT
33
2099
|
|
1132162
N/A
N/A
2222
2241
CATGAGATAATGCCATGAAA
50
2100
|
|
1132190
N/A
N/A
2329
2348
CCTAATACTGTTTGCTAGTT
36
2101
|
|
1132218
N/A
N/A
2464
2483
TATAGTGCCATCAGCTTAGC
43
2102
|
|
1132246
N/A
N/A
2579
2598
TTTGGCTTTTGGAAATGGCC
29
2103
|
|
1132274
N/A
N/A
2715
2734
AAAAATTTGCACATTCCCAG
85
2104
|
|
1132302
N/A
N/A
2820
2839
TGGTTCTACATTGAGAATGT
76
2105
|
|
1132330
N/A
N/A
2926
2945
ATACTTGTTGAATGTTGAAG
52
2106
|
|
1132358
N/A
N/A
3030
3049
CTGGAGTAGACTCCTGACTC
62
2107
|
|
1132386
N/A
N/A
3424
3443
TCAGGAAAAGGACAGACCCT
36
2108
|
|
1132414
N/A
N/A
3514
3533
CCCCGGTGACAGTGAGAACT
33
2109
|
|
1132442
N/A
N/A
3709
3728
GCCCACCCTGCCCTATCACA
40
2110
|
|
1132470
N/A
N/A
3877
3896
GTAATGAGGCGGGAGGAGAG
46
2111
|
|
1132498
N/A
N/A
3999
4018
CCTGCAAAATGGGACCACTC
60
2112
|
|
1132526
N/A
N/A
4105
4124
GAAGGAAGAGCAGGGAGCTG
58
2113
|
|
1132554
N/A
N/A
4295
4314
GTAGGCCCAGGGTTGCCCCT
71
2114
|
|
1132582
N/A
N/A
4523
4542
TTCCCAACCATCTGCTCACC
72
2115
|
|
1132610
N/A
N/A
4620
4639
CACCCCTGGGCCTAAAGACC
79
2116
|
|
1132638
N/A
N/A
4841
4860
CTCTCATCTGCTTTCCGCAC
60
2117
|
|
1132666
N/A
N/A
5158
5177
CGTTGTCCGGGTTCCTGTAG
59
2118
|
|
1132694
N/A
N/A
5708
5727
GGTACTCGCCGGTCCTGCAG
37
2119
|
|
1132722
N/A
N/A
6111
6130
TCTCTTCCCGTCCCCGCGGG
60
2120
|
|
1132750
N/A
N/A
6361
6380
CTAACCCAGTGATCAAAGGT
75
2121
|
|
1132778
N/A
N/A
6463
6482
ATTCTACCTGGGATTCACCT
34
2122
|
|
1132806
N/A
N/A
6609
6628
CTGGCTCCCACACTAGCCCG
44
2123
|
|
1132834
N/A
N/A
6697
6716
AGGCTGAGGTCCAGAGAAAA
96
2124
|
|
1132862
N/A
N/A
6814
6833
GCCGCCCGGACGATGGACAA
97
2125
|
|
1132890
N/A
N/A
7094
7113
GTCGGAAACACGCAGCTCAG
90†
2126
|
|
TABLE 32
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
No.
|
|
1130762
28
47
28
47
CATCCGTCCGTTGGTCCAGC
46
2127
|
|
1130790
74
93
74
93
TCCAAGCTCACCAGCAGGAA
41
2128
|
|
1130818
132
151
499
518
TTCAGCTTTGTACTTATGCT
63
2129
|
|
1130846
166
185
3517
3536
GCTCCCCGGTGACAGTGAGA
16
2130
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
15
183
|
|
1130875
221
240
3572
3591
TTGTGGGTACATTTGTGGTA
31
2131
|
|
1130903
287
306
3794
3813
CCCCATCGCTGGTCCTGATC
20
2132
|
|
1130931
328
347
N/A
N/A
TGCTGCAGTGGTCTTTCACT
55
2133
|
|
1130959
401
420
4209
4228
TGTTGTGGACAGAGACAGTG
67
2134
|
|
1130987
443
462
N/A
N/A
GGCTCAAAGCACTTCTCTTT
15
2135
|
|
1131015
485
504
4430
4449
CTATACCATATCTCATTCTT
20
2136
|
|
1131043
529
548
4474
4493
CAGGACCCTTGCACTGGCAT
38
2137
|
|
1131071
561
580
N/A
N/A
GGCCTGGCTGGCCAGCCGCT
114
2138
|
|
1131099
601
620
4685
4704
CTAGGCAGCGACCCCCATGG
40
2139
|
|
1131127
648
667
4732
4751
TCCGGTGTAGCCCACCGGGC
104
2140
|
|
1131155
704
723
4933
4952
TAGCTGAGCCCGCGGCCATC
22
2141
|
|
1131183
759
778
4988
5007
CGAGGCCCACGGCTGACAGG
51
2142
|
|
1131211
816
835
5045
5064
GCCGCCCAGTCCCCAGTTCC
41
2143
|
|
1131239
896
915
5210
5229
TACTCCCAGCTCAGCCGGTC
49
2144
|
|
1131267
990
1009
5304
5323
CGGCTGCGCGGGCATGAGTG
71
2145
|
|
1131295
1067
1086
5486
5505
TCCCGCTTCGCCGGCAAGGC
36
2146
|
|
1131323
1110
1129
5529
5548
CCCGCAGCTCAGTGGGCCGT
33
2147
|
|
1131351
1147
1166
5566
5585
CGCGGGTCATCGAAGACAGA
39
2148
|
|
1131379
1221
1240
5640
5659
GCAGAAACTGTGGCCCCAGT
56
2149
|
|
1131407
1308
1327
5968
5987
CACCACCGTCAGATCCTCGG
40
2150
|
|
1131435
1376
1395
6036
6055
AAGCGGTAGGAGCGCACGGC
25
2151
|
|
1131463
1451
1470
6194
6213
CTGCCGTCCGCATCCTCCTG
22
2152
|
|
1131491
1542
1561
6285
6304
GCCGGCCACCTGGCAGAGCG
87
2153
|
|
1131519
1574
1593
N/A
N/A
TATTCCTCCGCCCCCTCGAA
316
2154
|
|
1131547
1617
1636
6915
6934
CAGGGAGAGGAACGGTACCT
116
2155
|
|
1131575
1743
1762
7131
7150
GTCCTCACACACCAGCGGGC
14†
79
|
|
1131603
1790
1809
7178
7197
CCCCAGCTGATGATGCCTTG
50
2156
|
|
1131631
1820
1839
7208
7227
CCTGGCTTGTTGCGGTCACC
38
85
|
|
1131659
1867
1886
7255
7274
TGTGCTCCCGGATCCAGGCC
42
2157
|
|
1131687
1911
1930
7299
7318
TCACCAAGGAGGGAAAGATG
17
2158
|
|
1131715
1947
1966
7335
7354
TGCCTTCCATGCCCCAGCCA
36
156
|
|
1131743
2001
2020
7389
7408
TCCTGCGCCATCCTGGCGCG
30
2159
|
|
1131799
N/A
N/A
169
188
ATGAGCAGAGGCCATGGCTC
99
2160
|
|
1131827
N/A
N/A
265
284
ATCCACCCAGTCTGGTTGTC
89
2161
|
|
1131855
N/A
N/A
356
375
GAGATTTCTTCCCAAGACCC
107
2162
|
|
1131883
N/A
N/A
446
465
GAGAGAAGAAGGCAGGGAAC
126
2163
|
|
1131911
N/A
N/A
594
613
CTAGACTGCCCTGAGACTGC
72
2164
|
|
1131939
N/A
N/A
681
700
AGCCCATACCCTGCCCTCAA
38
2165
|
|
1131967
N/A
N/A
850
869
CCTGGCCTCTGTGTGGCATG
63
2166
|
|
1131995
N/A
N/A
947
966
CGCCCAGAGTCACCCAGCTC
34
2167
|
|
1132023
N/A
N/A
1055
1074
GGCTGAATACCTCACATGGG
74
2168
|
|
1132051
N/A
N/A
1188
1207
TCTGGAAGAATGAAAGCAAT
62
2169
|
|
1132079
N/A
N/A
1566
1585
GTCAGGCCCACCAGGGTAAG
47
2170
|
|
1132107
N/A
N/A
1705
1724
CAAGGATAGGGCACCAAGTT
56
2171
|
|
1132135
N/A
N/A
2114
2133
CCAGAACTAGGACAGTAAAT
52
2172
|
|
1132163
N/A
N/A
2225
2244
CTTCATGAGATAATGCCATG
46
2173
|
|
1132191
N/A
N/A
2332
2351
GAACCTAATACTGTTTGCTA
27
2174
|
|
1132219
N/A
N/A
2467
2486
TGCTATAGTGCCATCAGCTT
60
2175
|
|
1132247
N/A
N/A
2582
2601
GGATTTGGCTTTTGGAAATG
42
2176
|
|
1132275
N/A
N/A
2718
2737
GTGAAAAATTTGCACATTCC
50
2177
|
|
1132303
N/A
N/A
2823
2842
CTCTGGTTCTACATTGAGAA
36
2178
|
|
1132331
N/A
N/A
2929
2948
ATAATACTTGTTGAATGTTG
39
2179
|
|
1132359
N/A
N/A
3033
3052
TAGCTGGAGTAGACTCCTGA
92
2180
|
|
1132387
N/A
N/A
3427
3446
TGGTCAGGAAAAGGACAGAC
21
2181
|
|
1132415
N/A
N/A
3605
3624
TAGTCTTACCAGGGCTGAGG
50
2182
|
|
1132443
N/A
N/A
3734
3753
CTTCCCTGCTCTACCCAGGG
56
2183
|
|
1132471
N/A
N/A
3880
3899
AGAGTAATGAGGCGGGAGGA
44
2184
|
|
1132499
N/A
N/A
4002
4021
CCACCTGCAAAATGGGACCA
29
2185
|
|
1132527
N/A
N/A
4110
4129
TGGTGGAAGGAAGAGCAGGG
90
2186
|
|
1132555
N/A
N/A
4298
4317
TCTGTAGGCCCAGGGTTGCC
73
2187
|
|
1132583
N/A
N/A
4526
4545
CCGTTCCCAACCATCTGCTC
42
2188
|
|
1132611
N/A
N/A
4623
4642
AGCCACCCCTGGGCCTAAAG
150
2189
|
|
1132639
N/A
N/A
4844
4863
TCCCTCTCATCTGCTTTCCG
26
2190
|
|
1132667
N/A
N/A
5374
5393
CCACTTCCTAACCTCCCGGG
129
2191
|
|
1132695
N/A
N/A
5711
5730
GCGGGTACTCGCCGGTCCTG
58
2192
|
|
1132723
N/A
N/A
6114
6133
AGCTCTCTTCCCGTCCCCGC
67
2193
|
|
1132751
N/A
N/A
6364
6383
CGCCTAACCCAGTGATCAAA
91
2194
|
|
1132779
N/A
N/A
6472
6491
GGCCCTGGGATTCTACCTGG
50
2195
|
|
1132807
N/A
N/A
6612
6631
AACCTGGCTCCCACACTAGC
51
2196
|
|
1132835
N/A
N/A
6703
6722
GCAAGGAGGCTGAGGTCCAG
39
2197
|
|
1132863
N/A
N/A
6817
6836
CTTGCCGCCCGGACGATGGA
50
2198
|
|
1132891
N/A
N/A
7097
7116
TGGGTCGGAAACACGCAGCT
115†
2199
|
|
TABLE 33
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
No.
|
|
1130763
30
49
30
49
GGCATCCGTCCGTTGGTCCA
21
2200
|
|
1130791
75
94
75
94
CTCCAAGCTCACCAGCAGGA
37
2201
|
|
1130819
133
152
500
519
CTTCAGCTTTGTACTTATGC
70
2202
|
|
1130847
168
187
3519
3538
GGGCTCCCCGGTGACAGTGA
33
2203
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
14
183
|
|
1130876
222
241
3573
3592
CTTGTGGGTACATTTGTGGT
28
2204
|
|
1130904
288
307
3795
3814
TCCCCATCGCTGGTCCTGAT
43
2205
|
|
1130932
329
348
N/A
N/A
TTGCTGCAGTGGTCTTTCAC
24
2206
|
|
1130960
402
421
4210
4229
GTGTTGTGGACAGAGACAGT
34
2207
|
|
1130988
444
463
N/A
N/A
AGGCTCAAAGCACTTCTCTT
23
2208
|
|
1131016
487
506
4432
4451
TTCTATACCATATCTCATTC
30
2209
|
|
1131044
530
549
4475
4494
TCAGGACCCTTGCACTGGCA
31
2210
|
|
1131072
563
582
N/A
N/A
CAGGCCTGGCTGGCCAGCCG
41
2211
|
|
1131100
603
622
4687
4706
CTCTAGGCAGCGACCCCCAT
22
2212
|
|
1131128
649
668
4733
4752
CTCCGGTGTAGCCCACCGGG
44
2213
|
|
1131156
706
725
4935
4954
GGTAGCTGAGCCCGCGGCCA
29
2214
|
|
1131184
760
779
4989
5008
CCGAGGCCCACGGCTGACAG
78
2215
|
|
1131212
819
838
5048
5067
GTGGCCGCCCAGTCCCCAGT
52
2216
|
|
1131240
897
916
5211
5230
GTACTCCCAGCTCAGCCGGT
33
2217
|
|
1131268
991
1010
5305
5324
CCGGCTGCGCGGGCATGAGT
29
2218
|
|
1131296
1068
1087
5487
5506
CTCCCGCTTCGCCGGCAAGG
35
2219
|
|
1131324
1113
1132
5532
5551
CTGCCCGCAGCTCAGTGGGC
53
2220
|
|
1131352
1149
1168
5568
5587
GACGCGGGTCATCGAAGACA
30
2221
|
|
1131380
1222
1241
5641
5660
CGCAGAAACTGTGGCCCCAG
31
2222
|
|
1131408
1309
1328
5969
5988
GCACCACCGTCAGATCCTCG
49
2223
|
|
1131436
1377
1396
6037
6056
CAAGCGGTAGGAGCGCACGG
33
2224
|
|
1131464
1452
1471
6195
6214
GCTGCCGTCCGCATCCTCCT
38
2225
|
|
1131492
1544
1563
6287
6306
CAGCCGGCCACCTGGCAGAG
63
2226
|
|
1131520
1575
1594
N/A
N/A
ATATTCCTCCGCCCCCTCGA
216
2227
|
|
1131548
1618
1637
6916
6935
CCAGGGAGAGGAACGGTACC
119
2228
|
|
1131604
1791
1810
7179
7198
TCCCCAGCTGATGATGCCTT
29
2229
|
|
1131632
1821
1840
7209
7228
GCCTGGCTTGTTGCGGTCAC
27
2230
|
|
1131660
1868
1887
7256
7275
GTGTGCTCCCGGATCCAGGC
43
2231
|
|
1131688
1913
1932
7301
7320
AATCACCAAGGAGGGAAAGA
14
2232
|
|
1131716
1948
1967
7336
7355
TTGCCTTCCATGCCCCAGCC
19
2233
|
|
1131744
2003
2022
7391
7410
GTTCCTGCGCCATCCTGGCG
7
2234
|
|
1131772
N/A
N/A
88
107
CCGAAAGTGTTGACTCCAAG
64
2235
|
|
1131800
N/A
N/A
172
191
GTCATGAGCAGAGGCCATGG
43
2236
|
|
1131828
N/A
N/A
268
287
TGTATCCACCCAGTCTGGTT
70
2237
|
|
1131856
N/A
N/A
359
378
GCAGAGATTTCTTCCCAAGA
53
2238
|
|
1131884
N/A
N/A
451
470
CAAGGGAGAGAAGAAGGCAG
75
2239
|
|
1131912
N/A
N/A
597
616
AGACTAGACTGCCCTGAGAC
40
2240
|
|
1131940
N/A
N/A
684
703
CACAGCCCATACCCTGCCCT
57
2241
|
|
1131968
N/A
N/A
853
872
GGCCCTGGCCTCTGTGTGGC
70
2242
|
|
1131996
N/A
N/A
951
970
CCCTCGCCCAGAGTCACCCA
58
2243
|
|
1132024
N/A
N/A
1058
1077
ATGGGCTGAATACCTCACAT
44
2244
|
|
1132052
N/A
N/A
1191
1210
GTGTCTGGAAGAATGAAAGC
45
2245
|
|
1132080
N/A
N/A
1569
1588
CAAGTCAGGCCCACCAGGGT
47
2246
|
|
1132108
N/A
N/A
1708
1727
AGGCAAGGATAGGGCACCAA
44
2247
|
|
1132136
N/A
N/A
2117
2136
AAACCAGAACTAGGACAGTA
52
2248
|
|
1132164
N/A
N/A
2228
2247
GGGCTTCATGAGATAATGCC
63
2249
|
|
1132192
N/A
N/A
2335
2354
CAGGAACCTAATACTGTTTG
66
2250
|
|
1132220
N/A
N/A
2470
2489
GTGTGCTATAGTGCCATCAG
44
2251
|
|
1132248
N/A
N/A
2601
2620
CACTTTGTGCTTTTGCTGGG
59
2252
|
|
1132276
N/A
N/A
2721
2740
GCAGTGAAAAATTTGCACAT
41
2253
|
|
1132304
N/A
N/A
2826
2845
CTTCTCTGGTTCTACATTGA
52
2254
|
|
1132332
N/A
N/A
2933
2952
CTTAATAATACTTGTTGAAT
56
2255
|
|
1132360
N/A
N/A
3036
3055
AGATAGCTGGAGTAGACTCC
57
2256
|
|
1132388
N/A
N/A
3430
3449
GTCTGGTCAGGAAAAGGACA
74
2257
|
|
1132416
N/A
N/A
3625
3644
CCTGCTCCAACTCCTCTGCG
27
2258
|
|
1132444
N/A
N/A
3737
3756
AGGCTTCCCTGCTCTACCCA
61
2259
|
|
1132472
N/A
N/A
3883
3902
AGGAGAGTAATGAGGCGGGA
46
2260
|
|
1132500
N/A
N/A
4009
4028
GGTTTACCCACCTGCAAAAT
36
2261
|
|
1132528
N/A
N/A
4113
4132
ACATGGTGGAAGGAAGAGCA
35
2262
|
|
1132556
N/A
N/A
4301
4320
TATTCTGTAGGCCCAGGGTT
71
2263
|
|
1132584
N/A
N/A
4529
4548
GGCCCGTTCCCAACCATCTG
43
2264
|
|
1132612
N/A
N/A
4626
4645
GTGAGCCACCCCTGGGCCTA
80
2265
|
|
1132640
N/A
N/A
4848
4867
TGCCTCCCTCTCATCTGCTT
74
2266
|
|
1132668
N/A
N/A
5377
5396
CCCCCACTTCCTAACCTCCC
60
2267
|
|
1132696
N/A
N/A
5745
5764
GAGGAGCCGCGGCCCCTGGG
70
2268
|
|
1132724
N/A
N/A
6117
6136
CCAAGCTCTCTTCCCGTCCC
60
2269
|
|
1132752
N/A
N/A
6367
6386
TTCCGCCTAACCCAGTGATC
68
2270
|
|
1132780
N/A
N/A
6475
6494
CCAGGCCCTGGGATTCTACC
46
2271
|
|
1132808
N/A
N/A
6615
6634
CAGAACCTGGCTCCCACACT
83
2272
|
|
1132836
N/A
N/A
6706
6725
TAGGCAAGGAGGCTGAGGTC
80
2273
|
|
1132864
N/A
N/A
6820
6839
ACGCTTGCCGCCCGGACGAT
83
2274
|
|
1132892
N/A
N/A
7100
7119
CCCTGGGTCGGAAACACGCA
50†
2275
|
|
TABLE 34
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
No.
|
|
1130764
31
50
31
50
TGGCATCCGTCCGTTGGTCC
45
2276
|
|
1130792
77
96
77
96
GACTCCAAGCTCACCAGCAG
69
2277
|
|
1130820
134
153
501
520
TCTTCAGCTTTGTACTTATG
43
2278
|
|
1130848
169
188
3520
3539
AGGGCTCCCCGGTGACAGTG
30
2279
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
17
183
|
|
1130877
224
243
3575
3594
CCCTTGTGGGTACATTTGTG
45
2280
|
|
1130905
289
308
3796
3815
ATCCCCATCGCTGGTCCTGA
48
2281
|
|
1130933
330
349
N/A
N/A
TTTGCTGCAGTGGTCTTTCA
37
2282
|
|
1130961
403
422
4211
4230
GGTGTTGTGGACAGAGACAG
43
2283
|
|
1130989
445
464
4390
4409
GAGGCTCAAAGCACTTCTCT
32
2284
|
|
1131017
488
507
4433
4452
GTTCTATACCATATCTCATT
37
2285
|
|
1131045
531
550
4476
4495
ATCAGGACCCTTGCACTGGC
38
2286
|
|
1131073
564
583
N/A
N/A
GCAGGCCTGGCTGGCCAGCC
50
2287
|
|
1131101
604
623
4688
4707
CCTCTAGGCAGCGACCCCCA
41
2288
|
|
1131129
651
670
4735
4754
GGCTCCGGTGTAGCCCACCG
60
2289
|
|
1131157
707
726
4936
4955
CGGTAGCTGAGCCCGCGGCC
55
2290
|
|
1131185
761
780
4990
5009
TCCGAGGCCCACGGCTGACA
49
2291
|
|
1131213
836
855
N/A
N/A
GGGTTCCGGCAGAAGGCGTG
66
2292
|
|
1131241
899
918
5213
5232
CAGTACTCCCAGCTCAGCCG
38
2293
|
|
1131269
992
1011
5306
5325
GCCGGCTGCGCGGGCATGAG
51
2294
|
|
1131297
1070
1089
5489
5508
TGCTCCCGCTTCGCCGGCAA
30
2295
|
|
1131325
1115
1134
5534
5553
CGCTGCCCGCAGCTCAGTGG
45
2296
|
|
1131353
1150
1169
5569
5588
CGACGCGGGTCATCGAAGAC
67
2297
|
|
1131381
1223
1242
5642
5661
GCGCAGAAACTGTGGCCCCA
47
112
|
|
1131409
1310
1329
5970
5989
AGCACCACCGTCAGATCCTC
18
2298
|
|
1131437
1378
1397
6038
6057
GCAAGCGGTAGGAGCGCACG
50
2299
|
|
1131465
1455
1474
6198
6217
GCAGCTGCCGTCCGCATCCT
45
70
|
|
1131493
1547
1566
6290
6309
CCCCAGCCGGCCACCTGGCA
74
2300
|
|
1131521
1577
1596
N/A
N/A
GCATATTCCTCCGCCCCCTC
148
2301
|
|
1131549
1619
1638
6917
6936
TCCAGGGAGAGGAACGGTAC
130
2302
|
|
1131577
1745
1764
7133
7152
TGGTCCTCACACACCAGCGG
6†
2303
|
|
1131605
1792
1811
7180
7199
ATCCCCAGCTGATGATGCCT
28
2304
|
|
1131633
1822
1841
7210
7229
CGCCTGGCTTGTTGCGGTCA
34
2305
|
|
1131661
1869
1888
7257
7276
GGTGTGCTCCCGGATCCAGG
39
2306
|
|
1131689
1914
1933
7302
7321
GAATCACCAAGGAGGGAAAG
21
2307
|
|
1131717
1949
1968
7337
7356
CTTGCCTTCCATGCCCCAGC
24
2308
|
|
1131745
2004
2023
7392
7411
AGTTCCTGCGCCATCCTGGC
25
2309
|
|
1131773
N/A
N/A
91
110
TCACCGAAAGTGTTGACTCC
87
2310
|
|
1131801
N/A
N/A
175
194
CAGGTCATGAGCAGAGGCCA
70
2311
|
|
1131829
N/A
N/A
271
290
CCTTGTATCCACCCAGTCTG
76
2312
|
|
1131857
N/A
N/A
365
384
AGAAATGCAGAGATTTCTTC
91
2313
|
|
1131885
N/A
N/A
454
473
CTACAAGGGAGAGAAGAAGG
88
2314
|
|
1131913
N/A
N/A
600
619
ACTAGACTAGACTGCCCTGA
77
2315
|
|
1131941
N/A
N/A
687
706
AGACACAGCCCATACCCTGC
41
2316
|
|
1131969
N/A
N/A
856
875
GAAGGCCCTGGCCTCTGTGT
31
2317
|
|
1131997
N/A
N/A
954
973
AATCCCTCGCCCAGAGTCAC
90
2318
|
|
1132025
N/A
N/A
1061
1080
GTGATGGGCTGAATACCTCA
49
2319
|
|
1132053
N/A
N/A
1194
1213
TTAGTGTCTGGAAGAATGAA
60
2320
|
|
1132081
N/A
N/A
1572
1591
CACCAAGTCAGGCCCACCAG
80
2321
|
|
1132109
N/A
N/A
1711
1730
ATTAGGCAAGGATAGGGCAC
32
2322
|
|
1132137
N/A
N/A
2120
2139
ACCAAACCAGAACTAGGACA
56
2323
|
|
1132165
N/A
N/A
2250
2269
CTTCTTCAGAGGAATTGTCG
59
2324
|
|
1132193
N/A
N/A
2338
2357
TCACAGGAACCTAATACTGT
61
2325
|
|
1132221
N/A
N/A
2473
2492
GGTGTGTGCTATAGTGCCAT
59
2326
|
|
1132249
N/A
N/A
2604
2623
TCACACTTTGTGCTTTTGCT
35
2327
|
|
1132277
N/A
N/A
2724
2743
AGAGCAGTGAAAAATTTGCA
44
2328
|
|
1132305
N/A
N/A
2829
2848
ATTCTTCTCTGGTTCTACAT
56
2329
|
|
1132333
N/A
N/A
2936
2955
GCACTTAATAATACTTGTTG
41
2330
|
|
1132361
N/A
N/A
3039
3058
GTCAGATAGCTGGAGTAGAC
36
2331
|
|
1132389
N/A
N/A
3433
3452
AGGGTCTGGTCAGGAAAAGG
47
2332
|
|
1132417
N/A
N/A
3628
3647
GCCCCTGCTCCAACTCCTCT
63
2333
|
|
1132445
N/A
N/A
3740
3759
ACAAGGCTTCCCTGCTCTAC
102
2334
|
|
1132473
N/A
N/A
3886
3905
ACCAGGAGAGTAATGAGGCG
47
2335
|
|
1132501
N/A
N/A
4012
4031
CTTGGTTTACCCACCTGCAA
59
2336
|
|
1132529
N/A
N/A
4116
4135
TGGACATGGTGGAAGGAAGA
94
2337
|
|
1132557
N/A
N/A
4304
4323
ACCTATTCTGTAGGCCCAGG
53
2338
|
|
1132585
N/A
N/A
4532
4551
CCTGGCCCGTTCCCAACCAT
64
2339
|
|
1132613
N/A
N/A
4629
4648
GCAGTGAGCCACCCCTGGGC
72
2340
|
|
1132641
N/A
N/A
4851
4870
TCCTGCCTCCCTCTCATCTG
66
2341
|
|
1132669
N/A
N/A
5380
5399
CCCCCCCCACTTCCTAACCT
84
2342
|
|
1132697
N/A
N/A
5748
5767
ACGGAGGAGCCGCGGCCCCT
52
2343
|
|
1132725
N/A
N/A
6120
6139
GCCCCAAGCTCTCTTCCCGT
120
2344
|
|
1132753
N/A
N/A
6370
6389
TTCTTCCGCCTAACCCAGTG
71
2345
|
|
1132781
N/A
N/A
6478
6497
ATCCCAGGCCCTGGGATTCT
79
2346
|
|
1132809
N/A
N/A
6618
6637
TCGCAGAACCTGGCTCCCAC
60
2347
|
|
1132837
N/A
N/A
6709
6728
TCATAGGCAAGGAGGCTGAG
78
2348
|
|
1132865
N/A
N/A
6836
6855
CACACCCCATCTGACAACGC
82
2349
|
|
1132893
N/A
N/A
7103
7122
TCACCCTGGGTCGGAAACAC
73†
2350
|
|
TABLE 35
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130793
78
97
78
97
TGACTCCAAGCTCACCAGCA
34
239
|
|
1130821
135
154
502
521
CTCTTCAGCTTTGTACTTAT
42
240
|
|
1130849
171
190
3522
3541
GCAGGGCTCCCCGGTGACAG
34
241
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
11
183
|
|
1130878
225
244
3576
3595
GCCCTTGTGGGTACATTTGT
33
242
|
|
1130906
290
309
3797
3816
TATCCCCATCGCTGGTCCTG
26
243
|
|
1130934
331
350
N/A
N/A
GTTTGCTGCAGTGGTCTTTC
21
244
|
|
1130962
404
423
4212
4231
AGGTGTTGTGGACAGAGACA
45
245
|
|
1130990
446
465
4391
4410
TGAGGCTCAAAGCACTTCTC
18
246
|
|
1131018
489
508
4434
4453
AGTTCTATACCATATCTCAT
13
247
|
|
1131046
532
551
4477
4496
CATCAGGACCCTTGCACTGG
26
248
|
|
1131074
566
585
N/A
N/A
CGGCAGGCCTGGCTGGCCAG
70
249
|
|
1131102
606
625
4690
4709
CACCTCTAGGCAGCGACCCC
28
250
|
|
1131130
653
672
4737
4756
AAGGCTCCGGTGTAGCCCAC
66
251
|
|
1131158
709
728
4938
4957
CGCGGTAGCTGAGCCCGCGG
98
252
|
|
1131186
762
781
4991
5010
CTCCGAGGCCCACGGCTGAC
40
253
|
|
1131214
839
858
N/A
N/A
TCCGGGTTCCGGCAGAAGGC
39
55
|
|
1131242
902
921
5216
5235
TCGCAGTACTCCCAGCTCAG
16
254
|
|
1131270
993
1012
5307
5326
TGCCGGCTGCGCGGGCATGA
41
255
|
|
1131298
1071
1090
5490
5509
CTGCTCCCGCTTCGCCGGCA
27
256
|
|
1131326
1116
1135
5535
5554
CCGCTGCCCGCAGCTCAGTG
34
257
|
|
1131354
1152
1171
5571
5590
AACGACGCGGGTCATCGAAG
36
258
|
|
1131382
1224
1243
5643
5662
GGCGCAGAAACTGTGGCCCC
59
259
|
|
1131410
1311
1330
5971
5990
GAGCACCACCGTCAGATCCT
18
260
|
|
1131438
1380
1399
6040
6059
GTGCAAGCGGTAGGAGCGCA
40
261
|
|
1131466
1459
1478
6202
6221
GCGCGCAGCTGCCGTCCGCA
36
262
|
|
1131494
1548
1567
6291
6310
GCCCCAGCCGGCCACCTGGC
62
263
|
|
1131522
1578
1597
N/A
N/A
GGCATATTCCTCCGCCCCCT
79
264
|
|
1131550
1620
1639
6918
6937
CTCCAGGGAGAGGAACGGTA
74
265
|
|
1131578
1746
1765
7134
7153
TTGGTCCTCACACACCAGCG
8†
266
|
|
1131606
1793
1812
7181
7200
GATCCCCAGCTGATGATGCC
31
267
|
|
1131634
1823
1842
7211
7230
ACGCCTGGCTTGTTGCGGTC
31
268
|
|
1131662
1870
1889
7258
7277
CGGTGTGCTCCCGGATCCAG
22
269
|
|
1131690
1915
1934
7303
7322
GGAATCACCAAGGAGGGAAA
20
270
|
|
1131718
1950
1969
7338
7357
TCTTGCCTTCCATGCCCCAG
17
271
|
|
1131746
2005
2024
7393
7412
GAGTTCCTGCGCCATCCTGG
7
159
|
|
1131774
N/A
N/A
94
113
CACTCACCGAAAGTGTTGAC
78
272
|
|
1131802
N/A
N/A
178
197
CCACAGGTCATGAGCAGAGG
58
273
|
|
1131830
N/A
N/A
274
293
TGCCCTTGTATCCACCCAGT
97
274
|
|
1131858
N/A
N/A
368
387
CCCAGAAATGCAGAGATTTC
98
275
|
|
1131886
N/A
N/A
457
476
AATCTACAAGGGAGAGAAGA
79
276
|
|
1131914
N/A
N/A
603
622
GGCACTAGACTAGACTGCCC
67
?77
|
|
1131942
N/A
N/A
690
709
CTTAGACACAGCCCATACCC
60
278
|
|
1131970
N/A
N/A
859
878
ATTGAAGGCCCTGGCCTCTG
62
279
|
|
1131998
N/A
N/A
957
976
TCAAATCCCTCGCCCAGAGT
81
280
|
|
1132026
N/A
N/A
1064
1083
CAGGTGATGGGCTGAATACC
81
281
|
|
1132054
N/A
N/A
1197
1216
AATTTAGTGTCTGGAAGAAT
70
282
|
|
1132082
N/A
N/A
1575
1594
CCCCACCAAGTCAGGCCCAC
78
283
|
|
1132110
N/A
N/A
1747
1766
TGGACGGACAGAGAGGAGGG
71
284
|
|
1132138
N/A
N/A
2123
2142
AGGACCAAACCAGAACTAGG
58
285
|
|
1132166
N/A
N/A
2253
2272
TGTCTTCTTCAGAGGAATTG
66
286
|
|
1132194
N/A
N/A
2341
2360
GGCTCACAGGAACCTAATAC
47
287
|
|
1132222
N/A
N/A
2479
2498
GATTCAGGTGTGTGCTATAG
53
288
|
|
1132250
N/A
N/A
2609
2628
GTTTTTCACACTTTGTGCTT
51
289
|
|
1132278
N/A
N/A
2734
2753
GGGCATGCACAGAGCAGTGA
52
290
|
|
1132306
N/A
N/A
2832
2851
CTCATTCTTCTCTGGTTCTA
48
291
|
|
1132334
N/A
N/A
2939
2958
TAGGCACTTAATAATACTTG
56
292
|
|
1132362
N/A
N/A
3042
3061
GGAGTCAGATAGCTGGAGTA
69
293
|
|
1132390
N/A
N/A
3436
3455
CTCAGGGTCTGGTCAGGAAA
70
294
|
|
1132418
N/A
N/A
3631
3650
CAGGCCCCTGCTCCAACTCC
65
295
|
|
1132446
N/A
N/A
3743
3762
GAGACAAGGCTTCCCTGCTC
71
296
|
|
1132474
N/A
N/A
3889
3908
GATACCAGGAGAGTAATGAG
55
297
|
|
1132502
N/A
N/A
4015
4034
AAGCTTGGTTTACCCACCTG
46
298
|
|
1132530
N/A
N/A
4122
4141
GAGAGATGGACATGGTGGAA
67
299
|
|
1132558
N/A
N/A
4310
4329
CCAGCAACCTATTCTGTAGG
56
300
|
|
1132586
N/A
N/A
4535
4554
CTCCCTGGCCCGTTCCCAAC
68
301
|
|
1132614
N/A
N/A
4632
4651
AACGCAGTGAGCCACCCCTG
32
302
|
|
1132642
N/A
N/A
4859
4878
CTGGGCTCTCCTGCCTCCCT
31
303
|
|
1132670
N/A
N/A
5383
5402
TTCCCCCCCCCACTTCCTAA
106
304
|
|
1132698
N/A
N/A
5751
5770
GAGACGGAGGAGCCGCGGCC
72
305
|
|
1132726
N/A
N/A
6158
6177
AACCCGGGCGGAGAGGAGCG
98
306
|
|
1132754
N/A
N/A
6373
6392
GGCTTCTTCCGCCTAACCCA
72
307
|
|
1132782
N/A
N/A
6481
6500
TGAATCCCAGGCCCTGGGAT
101
308
|
|
1132810
N/A
N/A
6621
6640
GAGTCGCAGAACCTGGCTCC
71
309
|
|
1132838
N/A
N/A
6712
6731
ATTTCATAGGCAAGGAGGCT
66
310
|
|
1132866
N/A
N/A
6839
6858
CTTCACACCCCATCTGACAA
108
311
|
|
1132894
N/A
N/A
7106
7125
GAATCACCCTGGGTCGGAAA
76†
312
|
|
TABLE 36
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130794
79
98
79
98
TTGACTCCAAGCTCACCAGC
41
315
|
|
1130822
136
155
503
522
GCTCTTCAGCTTTGTACTTA
34
316
|
|
1130850
172
191
3523
3542
GGCAGGGCTCCCCGGTGACA
19
317
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
12
183
|
|
1130879
226
245
3577
3596
GGCCCTTGTGGGTACATTTG
66
318
|
|
1130907
292
311
3799
3818
AGTATCCCCATCGCTGGTCC
19
319
|
|
1130935
332
351
N/A
N/A
TGTTTGCTGCAGTGGTCTTT
27
320
|
|
1130963
405
424
4213
4232
GAGGTGTTGTGGACAGAGAC
47
321
|
|
1130991
447
466
4392
4411
CTGAGGCTCAAAGCACTTCT
29
322
|
|
1131019
493
512
4438
4457
GCTCAGTTCTATACCATATC
17
323
|
|
1131047
533
552
4478
4497
GCATCAGGACCCTTGCACTG
26
324
|
|
1131075
568
587
N/A
N/A
TGCGGCAGGCCTGGCTGGCC
50
325
|
|
1131103
607
626
4691
4710
CCACCTCTAGGCAGCGACCC
29
326
|
|
1131131
654
673
4738
4757
GAAGGCTCCGGTGTAGCCCA
56
327
|
|
1131159
710
729
4939
4958
CCGCGGTAGCTGAGCCCGCG
77
328
|
|
1131187
763
782
4992
5011
CCTCCGAGGCCCACGGCTGA
47
329
|
|
1131215
840
859
N/A
N/A
GTCCGGGTTCCGGCAGAAGG
38
313
|
|
1131243
903
922
5217
5236
GTCGCAGTACTCCCAGCTCA
12
330
|
|
1131271
994
1013
5308
5327
GTGCCGGCTGCGCGGGCATG
63
331
|
|
1131299
1072
1091
5491
5510
GCTGCTCCCGCTTCGCCGGC
26
332
|
|
1131327
1117
1136
5536
5555
GCCGCTGCCCGCAGCTCAGT
30
333
|
|
1131355
1153
1172
5572
5591
CAACGACGCGGGTCATCGAA
53
334
|
|
1131383
1225
1244
5644
5663
CGGCGCAGAAACTGTGGCCC
46
335
|
|
1131411
1312
1331
5972
5991
CGAGCACCACCGTCAGATCC
17
336
|
|
1131439
1381
1400
6041
6060
CGTGCAAGCGGTAGGAGCGC
27
337
|
|
1131467
1460
1479
6203
6222
AGCGCGCAGCTGCCGTCCGC
42
338
|
|
1131495
1549
1568
6292
6311
GGCCCCAGCCGGCCACCTGG
82
339
|
|
1131523
1579
1598
N/A
N/A
TGGCATATTCCTCCGCCCCC
67
340
|
|
1131551
1621
1640
6919
6938
GCTCCAGGGAGAGGAACGGT
66
341
|
|
1131579
1747
1766
7135
7154
CTTGGTCCTCACACACCAGC
36†
342
|
|
1131607
1794
1813
7182
7201
CGATCCCCAGCTGATGATGC
41
343
|
|
1131635
1839
1858
7227
7246
GGCCACATCGGTGTAGACGC
18
86
|
|
1131663
1871
1890
7259
7278
ACGGTGTGCTCCCGGATCCA
16
344
|
|
1131691
1916
1935
7304
7323
CGGAATCACCAAGGAGGGAA
15
345
|
|
1131719
1951
1970
7339
7358
ATCTTGCCTTCCATGCCCCA
28
346
|
|
1131747
2006
2025
7394
7413
TGAGTTCCTGCGCCATCCTG
17
347
|
|
1131775
N/A
N/A
97
116
CAGCACTCACCGAAAGTGTT
84
348
|
|
1131803
N/A
N/A
181
200
GACCCACAGGTCATGAGCAG
87
349
|
|
1131831
N/A
N/A
277
296
CTGTGCCCTTGTATCCACCC
61
350
|
|
1131859
N/A
N/A
371
390
CAGCCCAGAAATGCAGAGAT
69
351
|
|
1131887
N/A
N/A
460
479
TGGAATCTACAAGGGAGAGA
89
352
|
|
1131915
N/A
N/A
606
625
GTAGGCACTAGACTAGACTG
52
353
|
|
1131943
N/A
N/A
693
712
GCACTTAGACACAGCCCATA
64
354
|
|
1131971
N/A
N/A
862
881
CCCATTGAAGGCCCTGGCCT
49
355
|
|
1131999
N/A
N/A
960
979
AACTCAAATCCCTCGCCCAG
59
356
|
|
1132027
N/A
N/A
1067
1086
CACCAGGTGATGGGCTGAAT
49
357
|
|
1132055
N/A
N/A
1200
1219
GGTAATTTAGTGTCTGGAAG
46
358
|
|
1132083
N/A
N/A
1578
1597
CAACCCCACCAAGTCAGGCC
69
359
|
|
1132111
N/A
N/A
1750
1769
GGATGGACGGACAGAGAGGA
63
360
|
|
1132139
N/A
N/A
2126
2145
CCAAGGACCAAACCAGAACT
56
361
|
|
1132167
N/A
N/A
2256
2275
CTGTGTCTTCTTCAGAGGAA
50
362
|
|
1132195
N/A
N/A
2370
2389
TGTTGGTTGATGAAAATGTT
54
363
|
|
1132223
N/A
N/A
2482
2501
CTTGATTCAGGTGTGTGCTA
50
364
|
|
1132251
N/A
N/A
2628
2647
TCAGTCTACTTAGTGCAACG
51
365
|
|
1132279
N/A
N/A
2739
2758
TTTGTGGGCATGCACAGAGC
53
366
|
|
1132307
N/A
N/A
2835
2854
ATCCTCATTCTTCTCTGGTT
53
367
|
|
1132335
N/A
N/A
2942
2961
AAGTAGGCACTTAATAATAC
67
368
|
|
1132363
N/A
N/A
3045
3064
TCAGGAGTCAGATAGCTGGA
77
369
|
|
1132391
N/A
N/A
3439
3458
TCCCTCAGGGTCTGGTCAGG
54
370
|
|
1132419
N/A
N/A
3637
3656
GTCTCCCAGGCCCCTGCTCC
56
371
|
|
1132447
N/A
N/A
3749
3768
TAGAAAGAGACAAGGCTTCC
61
372
|
|
1132475
N/A
N/A
3892
3911
GGTGATACCAGGAGAGTAAT
47
373
|
|
1132503
N/A
N/A
4018
4037
TCCAAGCTTGGTTTACCCAC
38
374
|
|
1132531
N/A
N/A
4125
4144
TCTGAGAGATGGACATGGTG
68
375
|
|
1132559
N/A
N/A
4313
4332
TATCCAGCAACCTATTCTGT
69
376
|
|
1132587
N/A
N/A
4538
4557
CTCCTCCCTGGCCCGTTCCC
70
377
|
|
1132615
N/A
N/A
4635
4654
GGGAACGCAGTGAGCCACCC
49
378
|
|
1132643
N/A
N/A
4862
4881
AGGCTGGGCTCTCCTGCCTC
75
379
|
|
1132671
N/A
N/A
5386
5405
TCCTTCCCCCCCCCACTTCC
92
380
|
|
1132699
N/A
N/A
5754
5773
TGGGAGACGGAGGAGCCGCG
61
381
|
|
1132727
N/A
N/A
6161
6180
GCTAACCCGGGCGGAGAGGA
101
382
|
|
1132755
N/A
N/A
6376
6395
GCGGGCTTCTTCCGCCTAAC
56
383
|
|
1132783
N/A
N/A
6484
6503
CAGTGAATCCCAGGCCCTGG
66
384
|
|
1132811
N/A
N/A
6624
6643
CCAGAGTCGCAGAACCTGGC
56
385
|
|
1132839
N/A
N/A
6715
6734
TCAATTTCATAGGCAAGGAG
64
386
|
|
1132867
N/A
N/A
6842
6861
CTTCTTCACACCCCATCTGA
88
387
|
|
1132895
N/A
N/A
7109
7128
CCGGAATCACCCTGGGTCGG
28†
388
|
|
TABLE 37
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130795
80
99
80
99
GTTGACTCCAAGCTCACCAG
40
389
|
|
1130823
138
157
505
524
GTGCTCTTCAGCTTTGTACT
38
390
|
|
1130851
174
193
3525
3544
GTGGCAGGGCTCCCCGGTGA
80
391
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
11
183
|
|
1130880
228
247
3579
3598
CCGGCCCTTGTGGGTACATT
42
392
|
|
1130908
293
312
3800
3819
CAGTATCCCCATCGCTGGTC
24
393
|
|
1130936
333
352
N/A
N/A
GTGTTTGCTGCAGTGGTCTT
32
394
|
|
1130964
406
425
4214
4233
TGAGGTGTTGTGGACAGAGA
112
395
|
|
1130992
449
468
4394
4413
AGCTGAGGCTCAAAGCACTT
45
396
|
|
1131020
494
513
4439
4458
TGCTCAGTTCTATACCATAT
23
397
|
|
1131048
535
554
4480
4499
GGGCATCAGGACCCTTGCAC
81
398
|
|
1131076
569
588
N/A
N/A
GTGCGGCAGGCCTGGCTGGC
54
399
|
|
1131104
609
628
4693
4712
CTCCACCTCTAGGCAGCGAC
45
400
|
|
1131132
655
674
4739
4758
AGAAGGCTCCGGTGTAGCCC
69
401
|
|
1131160
711
730
4940
4959
GCCGCGGTAGCTGAGCCCGC
97
402
|
|
1131188
765
784
4994
5013
GGCCTCCGAGGCCCACGGCT
45
403
|
|
1131216
843
862
N/A
N/A
GTTGTCCGGGTTCCGGCAGA
20
404
|
|
1131244
904
923
5218
5237
GGTCGCAGTACTCCCAGCTC
32
405
|
|
1131272
996
1015
5310
5329
CGGTGCCGGCTGCGCGGGCA
59
406
|
|
1131300
1073
1092
5492
5511
GGCTGCTCCCGCTTCGCCGG
50
407
|
|
1131328
1118
1137
5537
5556
AGCCGCTGCCCGCAGCTCAG
83
408
|
|
1131356
1155
1174
5574
5593
GCCAACGACGCGGGTCATCG
38
409
|
|
1131384
1227
1246
5646
5665
GCCGGCGCAGAAACTGTGGC
68
410
|
|
1131412
1313
1332
5973
5992
CCGAGCACCACCGTCAGATC
47
411
|
|
1131440
1399
1418
6059
6078
TGACGGGCGAGAAGGCCTCG
59
412
|
|
1131468
1462
1481
6205
6224
GGAGCGCGCAGCTGCCGTCC
44
413
|
|
1131496
1550
1569
6293
6312
TGGCCCCAGCCGGCCACCTG
96
414
|
|
1131524
1581
1600
6879
6898
GCTGGCATATTCCTCCGCCC
64
415
|
|
1131552
1622
1641
6920
6939
CGCTCCAGGGAGAGGAACGG
107
416
|
|
1131580
1748
1767
7136
7155
GCTTGGTCCTCACACACCAG
10†
80
|
|
1131608
1795
1814
7183
7202
CCGATCCCCAGCTGATGATG
28
417
|
|
1131636
1842
1861
7230
7249
GTAGGCCACATCGGTGTAGA
51
418
|
|
1131664
1873
1892
7261
7280
AAACGGTGTGCTCCCGGATC
81
419
|
|
1131692
1917
1936
7305
7324
GCGGAATCACCAAGGAGGGA
18
420
|
|
1131720
1952
1971
7340
7359
AATCTTGCCTTCCATGCCCC
30
421
|
|
1131748
2007
2026
7395
7414
TTGAGTTCCTGCGCCATCCT
11
422
|
|
1131776
N/A
N/A
100
119
CCACAGCACTCACCGAAAGT
116
423
|
|
1131804
N/A
N/A
184
203
CTGGACCCACAGGTCATGAG
48
424
|
|
1131832
N/A
N/A
280
299
AGGCTGTGCCCTTGTATCCA
45
425
|
|
1131860
N/A
N/A
374
393
TCACAGCCCAGAAATGCAGA
144
426
|
|
1131888
N/A
N/A
463
482
AGGTGGAATCTACAAGGGAG
72
427
|
|
1131916
N/A
N/A
609
628
CAGGTAGGCACTAGACTAGA
85
428
|
|
1131944
N/A
N/A
696
715
TGGGCACTTAGACACAGCCC
56
429
|
|
1131972
N/A
N/A
865
884
TTGCCCATTGAAGGCCCTGG
60
430
|
|
1132000
N/A
N/A
963
982
GAGAACTCAAATCCCTCGCC
77
431
|
|
1132028
N/A
N/A
1070
1089
ATGCACCAGGTGATGGGCTG
65
432
|
|
1132056
N/A
N/A
1203
1222
GTAGGTAATTTAGTGTCTGG
89
433
|
|
1132084
N/A
N/A
1581
1600
ACTCAACCCCACCAAGTCAG
75
434
|
|
1132112
N/A
N/A
1753
1772
ATGGGATGGACGGACAGAGA
60
435
|
|
1132140
N/A
N/A
2129
2148
TTGCCAAGGACCAAACCAGA
87
436
|
|
1132168
N/A
N/A
2259
2278
TGCCTGTGTCTTCTTCAGAG
51
437
|
|
1132196
N/A
N/A
2373
2392
TGCTGTTGGTTGATGAAAAT
51
438
|
|
1132224
N/A
N/A
2485
2504
ACACTTGATTCAGGTGTGTG
64
439
|
|
1132252
N/A
N/A
2634
2653
TCCTTCTCAGTCTACTTAGT
53
440
|
|
1132280
N/A
N/A
2742
2761
CCATTTGTGGGCATGCACAG
60
441
|
|
1132308
N/A
N/A
2838
2857
TTGATCCTCATTCTTCTCTG
44
442
|
|
1132336
N/A
N/A
2950
2969
TTGCCACAAAGTAGGCACTT
66
443
|
|
1132364
N/A
N/A
3048
3067
GGTTCAGGAGTCAGATAGCT
74
444
|
|
1132392
N/A
N/A
3442
3461
CTGTCCCTCAGGGTCTGGTC
74
445
|
|
1132420
N/A
N/A
3640
3659
CATGTCTCCCAGGCCCCTGC
49
446
|
|
1132448
N/A
N/A
3752
3771
CTGTAGAAAGAGACAAGGCT
52
447
|
|
1132476
N/A
N/A
3895
3914
TCTGGTGATACCAGGAGAGT
50
448
|
|
1132504
N/A
N/A
4021
4040
GTTTCCAAGCTTGGTTTACC
61
449
|
|
1132532
N/A
N/A
4128
4147
TGGTCTGAGAGATGGACATG
40
450
|
|
1132560
N/A
N/A
4316
4335
GAGTATCCAGCAACCTATTC
46
451
|
|
1132588
N/A
N/A
4541
4560
ACGCTCCTCCCTGGCCCGTT
85
452
|
|
1132616
N/A
N/A
4638
4657
GGAGGGAACGCAGTGAGCCA
83
453
|
|
1132644
N/A
N/A
4866
4885
GCCAAGGCTGGGCTCTCCTG
43
454
|
|
1132672
N/A
N/A
5389
5408
TCCTCCTTCCCCCCCCCACT
99
455
|
|
1132700
N/A
N/A
5757
5776
CGCTGGGAGACGGAGGAGCC
98
456
|
|
1132728
N/A
N/A
6164
6183
AGAGCTAACCCGGGCGGAGA
69
457
|
|
1132756
N/A
N/A
6379
6398
GTCGCGGGCTTCTTCCGCCT
46
458
|
|
1132784
N/A
N/A
6487
6506
CAGCAGTGAATCCCAGGCCC
63
459
|
|
1132812
N/A
N/A
6627
6646
CATCCAGAGTCGCAGAACCT
48
460
|
|
1132840
N/A
N/A
6718
6737
AATTCAATTTCATAGGCAAG
69
461
|
|
1132868
N/A
N/A
6845
6864
CGCCTTCTTCACACCCCATC
97
462
|
|
1132896
N/A
N/A
7112
7131
CCTCCGGAATCACCCTGGGT
53†
463
|
|
TABLE 38
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130796
82
101
82
101
GTGTTGACTCCAAGCTCACC
34
465
|
|
1130824
139
158
506
525
TGTGCTCTTCAGCTTTGTAC
21
466
|
|
1130852
175
194
3526
3545
AGTGGCAGGGCTCCCCGGTG
59
467
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
16
183
|
|
1130881
229
248
3580
3599
GCCGGCCCTTGTGGGTACAT
52
468
|
|
1130909
294
313
3801
3820
ACAGTATCCCCATCGCTGGT
46
469
|
|
1130937
334
353
4142
4161
TGTGTTTGCTGCAGTGGTCT
63
470
|
|
1130965
407
426
4215
4234
GTGAGGTGTTGTGGACAGAG
40
471
|
|
1130993
450
469
4395
4414
AAGCTGAGGCTCAAAGCACT
37
472
|
|
1131021
495
514
4440
4459
TTGCTCAGTTCTATACCATA
16
473
|
|
1131049
536
555
4481
4500
TGGGCATCAGGACCCTTGCA
47
474
|
|
1131077
570
589
N/A
N/A
GGTGCGGCAGGCCTGGCTGG
52
475
|
|
1131105
610
629
4694
4713
CCTCCACCTCTAGGCAGCGA
48
476
|
|
1131133
656
675
4740
4759
CAGAAGGCTCCGGTGTAGCC
89
477
|
|
1131161
712
731
4941
4960
GGCCGCGGTAGCTGAGCCCG
59
478
|
|
1131189
766
785
4995
5014
TGGCCTCCGAGGCCCACGGC
75
479
|
|
1131217
844
863
N/A
N/A
CGTTGTCCGGGTTCCGGCAG
32
480
|
|
1131245
905
924
5219
5238
AGGTCGCAGTACTCCCAGCT
37
481
|
|
1131273
997
1016
5311
5330
GCGGTGCCGGCTGCGCGGGC
49
482
|
|
1131301
1075
1094
5494
5513
GCGGCTGCTCCCGCTTCGCC
48
483
|
|
1131329
1120
1139
5539
5558
GGAGCCGCTGCCCGCAGCTC
51
484
|
|
1131357
1156
1175
5575
5594
CGCCAACGACGCGGGTCATC
23
485
|
|
1131385
1228
1247
5647
5666
TGCCGGCGCAGAAACTGTGG
61
486
|
|
1131413
1314
1333
5974
5993
GCCGAGCACCACCGTCAGAT
35
487
|
|
1131441
1400
1419
6060
6079
CTGACGGGCGAGAAGGCCTC
37
488
|
|
1131469
1463
1482
6206
6225
AGGAGCGCGCAGCTGCCGTC
77
464
|
|
1131497
1551
1570
6294
6313
GTGGCCCCAGCCGGCCACCT
114
489
|
|
1131525
1582
1601
6880
6899
AGCTGGCATATTCCTCCGCC
113
490
|
|
1131553
1623
1642
6921
6940
GCGCTCCAGGGAGAGGAACG
90
491
|
|
1131581
1749
1768
7137
7156
AGCTTGGTCCTCACACACCA
7†
492
|
|
1131609
1796
1815
7184
7203
CCCGATCCCCAGCTGATGAT
33
493
|
|
1131637
1843
1862
7231
7250
AGTAGGCCACATCGGTGTAG
37
494
|
|
1131665
1874
1893
7262
7281
GAAACGGTGTGCTCCCGGAT
36
495
|
|
1131693
1918
1937
7306
7325
TGCGGAATCACCAAGGAGGG
11
496
|
|
1131721
1954
1973
7342
7361
ACAATCTTGCCTTCCATGCC
25
497
|
|
1131749
2009
2028
7397
7416
TATTGAGTTCCTGCGCCATC
13
498
|
|
1131777
N/A
N/A
103
122
TTCCCACAGCACTCACCGAA
77
499
|
|
1131805
N/A
N/A
187
206
CACCTGGACCCACAGGTCAT
88
500
|
|
1131833
N/A
N/A
283
302
TGCAGGCTGTGCCCTTGTAT
115
501
|
|
1131861
N/A
N/A
377
396
CTCTCACAGCCCAGAAATGC
73
502
|
|
1131889
N/A
N/A
466
485
CCAAGGTGGAATCTACAAGG
30
503
|
|
1131917
N/A
N/A
612
631
CACCAGGTAGGCACTAGACT
42
504
|
|
1131945
N/A
N/A
716
735
CCTGCTCCGAGCCAGGCTCG
62
505
|
|
1131973
N/A
N/A
868
887
TCCTTGCCCATTGAAGGCCC
43
506
|
|
1132001
N/A
N/A
966
985
TTGGAGAACTCAAATCCCTC
52
507
|
|
1132029
N/A
N/A
1073
1092
TCCATGCACCAGGTGATGGG
54
508
|
|
1132057
N/A
N/A
1206
1225
CATGTAGGTAATTTAGTGTC
47
509
|
|
1132085
N/A
N/A
1584
1603
GAGACTCAACCCCACCAAGT
60
510
|
|
1132113
N/A
N/A
1756
1775
CAGATGGGATGGACGGACAG
74
511
|
|
1132141
N/A
N/A
2132
2151
CACTTGCCAAGGACCAAACC
69
512
|
|
1132169
N/A
N/A
2262
2281
AATTGCCTGTGTCTTCTTCA
51
513
|
|
1132197
N/A
N/A
2376
2395
ATATGCTGTTGGTTGATGAA
48
514
|
|
1132225
N/A
N/A
2488
2507
TAGACACTTGATTCAGGTGT
43
515
|
|
1132253
N/A
N/A
2637
2656
GTGTCCTTCTCAGTCTACTT
45
516
|
|
1132281
N/A
N/A
2745
2764
TGGCCATTTGTGGGCATGCA
69
517
|
|
1132309
N/A
N/A
2841
2860
CAGTTGATCCTCATTCTTCT
57
518
|
|
1132337
N/A
N/A
2953
2972
CACTTGCCACAAAGTAGGCA
44
519
|
|
1132365
N/A
N/A
3051
3070
TTGGGTTCAGGAGTCAGATA
73
520
|
|
1132393
N/A
N/A
3445
3464
GCACTGTCCCTCAGGGTCTG
47
521
|
|
1132421
N/A
N/A
3643
3662
GTACATGTCTCCCAGGCCCC
50
522
|
|
1132449
N/A
N/A
3755
3774
CACCTGTAGAAAGAGACAAG
58
523
|
|
1132477
N/A
N/A
3898
3917
GGGTCTGGTGATACCAGGAG
44
524
|
|
1132505
N/A
N/A
4024
4043
CAAGTTTCCAAGCTTGGTTT
29
525
|
|
1132533
N/A
N/A
4131
4150
CAGTGGTCTGAGAGATGGAC
46
526
|
|
1132561
N/A
N/A
4319
4338
TCCGAGTATCCAGCAACCTA
50
527
|
|
1132589
N/A
N/A
4557
4576
CCAGCCTGTCTTCCTGACGC
67
528
|
|
1132617
N/A
N/A
4657
4676
GTTGGTGCGGCAGGCTTGGG
59
529
|
|
1132645
N/A
N/A
4869
4888
GCAGCCAAGGCTGGGCTCTC
59
530
|
|
1132673
N/A
N/A
5392
5411
GGCTCCTCCTTCCCCCCCCC
41
531
|
|
1132701
N/A
N/A
5760
5779
CTGCGCTGGGAGACGGAGGA
70
532
|
|
1132729
N/A
N/A
6167
6186
AACAGAGCTAACCCGGGCGG
65
533
|
|
1132757
N/A
N/A
6382
6401
AAAGTCGCGGGCTTCTTCCG
31
534
|
|
1132785
N/A
N/A
6490
6509
TCCCAGCAGTGAATCCCAGG
69
535
|
|
1132813
N/A
N/A
6630
6649
ACCCATCCAGAGTCGCAGAA
43
536
|
|
1132841
N/A
N/A
6721
6740
ATTAATTCAATTTCATAGGC
68
537
|
|
1132869
N/A
N/A
6862
6881
CCCCTGCGAACACAGAGCGC
59
538
|
|
TABLE 39
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130797
83
102
83
102
AGTGTTGACTCCAAGCTCAC
27
541
|
|
1130825
141
160
508
527
TGTGTGCTCTTCAGCTTTGT
14
542
|
|
1130853
177
196
3528
3547
GAAGTGGCAGGGCTCCCCGG
33
543
|
|
1130871
216
235
3567
3586
GGTACATTTGTGGTACAGCT
9
183
|
|
1130882
231
250
3582
3601
TGGCCGGCCCTTGTGGGTAC
23
544
|
|
1130910
296
315
3803
3822
AAACAGTATCCCCATCGCTG
36
545
|
|
1130938
335
354
4143
4162
CTGTGTTTGCTGCAGTGGTC
13
546
|
|
1130966
408
427
4216
4235
AGTGAGGTGTTGTGGACAGA
48
547
|
|
1130994
451
470
4396
4415
GAAGCTGAGGCTCAAAGCAC
22
548
|
|
1131022
496
515
4441
4460
CTTGCTCAGTTCTATACCAT
23
549
|
|
1131050
537
556
4482
4501
GTGGGCATCAGGACCCTTGC
41
550
|
|
1131078
571
590
N/A
N/A
TGGTGCGGCAGGCCTGGCTG
33
551
|
|
1131106
612
631
4696
4715
GCCCTCCACCTCTAGGCAGC
26
552
|
|
1131134
657
676
4741
4760
GCAGAAGGCTCCGGTGTAGC
23
553
|
|
1131162
714
733
4943
4962
CAGGCCGCGGTAGCTGAGCC
34
554
|
|
1131190
767
786
4996
5015
GTGGCCTCCGAGGCCCACGG
50
555
|
|
1131218
862
881
5176
5195
AGCACCACGGGCGGATGTCG
25
539
|
|
1131246
907
926
5221
5240
CCAGGTCGCAGTACTCCCAG
26
556
|
|
1131274
998
1017
5312
5331
GGCGGTGCCGGCTGCGCGGG
71
557
|
|
1131302
1076
1095
5495
5514
GGCGGCTGCTCCCGCTTCGC
20
558
|
|
1131330
1121
1140
5540
5559
CGGAGCCGCTGCCCGCAGCT
33
559
|
|
1131358
1158
1177
5577
5596
CCCGCCAACGACGCGGGTCA
19
560
|
|
1131386
1229
1248
5648
5667
CTGCCGGCGCAGAAACTGTG
46
561
|
|
1131414
1315
1334
5975
5994
GGCCGAGCACCACCGTCAGA
32
562
|
|
1131442
1401
1420
6061
6080
GCTGACGGGCGAGAAGGCCT
19
563
|
|
1131470
1464
1483
6207
6226
CAGGAGCGCGCAGCTGCCGT
33
564
|
|
1131498
1552
1571
6295
6314
GGTGGCCCCAGCCGGCCACC
46
565
|
|
1131526
1583
1602
6881
6900
AAGCTGGCATATTCCTCCGC
136
566
|
|
1131554
1657
1676
6955
6974
GGATGGAGGATCCGTGCACG
15†
567
|
|
1131582
1750
1769
7138
7157
CAGCTTGGTCCTCACACACC
11†
568
|
|
1131610
1797
1816
7185
7204
GCCCGATCCCCAGCTGATGA
39
569
|
|
1131638
1844
1863
7232
7251
TAGTAGGCCACATCGGTGTA
25
87
|
|
1131666
1876
1895
7264
7283
AGGAAACGGTGTGCTCCCGG
31
570
|
|
1131694
1919
1938
7307
7326
CTGCGGAATCACCAAGGAGG
10
571
|
|
1131722
1955
1974
7343
7362
CACAATCTTGCCTTCCATGC
18
572
|
|
1131750
2010
2029
7398
7417
TTATTGAGTTCCTGCGCCAT
23
573
|
|
1131778
N/A
N/A
106
125
TGGTTCCCACAGCACTCACC
42
574
|
|
1131806
N/A
N/A
190
209
AGTCACCTGGACCCACAGGT
28
575
|
|
1131834
N/A
N/A
286
305
AAATGCAGGCTGTGCCCTTG
33
576
|
|
1131862
N/A
N/A
380
399
TTCCTCTCACAGCCCAGAAA
41
577
|
|
1131890
N/A
N/A
469
488
TTCCCAAGGTGGAATCTACA
74
578
|
|
1131918
N/A
N/A
615
634
TAGCACCAGGTAGGCACTAG
45
579
|
|
1131946
N/A
N/A
721
740
AAGCACCTGCTCCGAGCCAG
33
580
|
|
1131974
N/A
N/A
871
890
CCTTCCTTGCCCATTGAAGG
46
581
|
|
1132002
N/A
N/A
969
988
AGCTTGGAGAACTCAAATCC
56
582
|
|
1132030
N/A
N/A
1076
1095
ATTTCCATGCACCAGGTGAT
58
583
|
|
1132058
N/A
N/A
1209
1228
TGGCATGTAGGTAATTTAGT
34
584
|
|
1132086
N/A
N/A
1587
1606
TTAGAGACTCAACCCCACCA
55
585
|
|
1132114
N/A
N/A
1759
1778
ATGCAGATGGGATGGACGGA
48
586
|
|
1132142
N/A
N/A
2135
2154
GTGCACTTGCCAAGGACCAA
25
587
|
|
1132170
N/A
N/A
2265
2284
GAGAATTGCCTGTGTCTTCT
40
588
|
|
1132198
N/A
N/A
2379
2398
ATTATATGCTGTTGGTTGAT
28
589
|
|
1132226
N/A
N/A
2513
2532
GCTACCTTAGGGAGAAAGCG
53
590
|
|
1132254
N/A
N/A
2640
2659
TGAGTGTCCTTCTCAGTCTA
47
591
|
|
1132282
N/A
N/A
2748
2767
TCATGGCCATTTGTGGGCAT
46
592
|
|
1132310
N/A
N/A
2845
2864
AGTACAGTTGATCCTCATTC
56
593
|
|
1132338
N/A
N/A
2956
2975
GAGCACTTGCCACAAAGTAG
30
594
|
|
1132366
N/A
N/A
3054
3073
AACTTGGGTTCAGGAGTCAG
42
595
|
|
1132394
N/A
N/A
3448
3467
CAGGCACTGTCCCTCAGGGT
32
596
|
|
1132422
N/A
N/A
3649
3668
GGCAGGGTACATGTCTCCCA
36
597
|
|
1132450
N/A
N/A
3758
3777
GCACACCTGTAGAAAGAGAC
39
598
|
|
1132478
N/A
N/A
3916
3935
CAGAATCCCAGGTGTGTGGG
64
599
|
|
1132506
N/A
N/A
4027
4046
CTCCAAGTTTCCAAGCTTGG
38
600
|
|
1132534
N/A
N/A
4134
4153
CTGCAGTGGTCTGAGAGATG
36
601
|
|
1132562
N/A
N/A
4325
4344
CAAGTCTCCGAGTATCCAGC
49
602
|
|
1132590
N/A
N/A
4560
4579
CTGCCAGCCTGTCTTCCTGA
30
603
|
|
1132618
N/A
N/A
4660
4679
CGGGTTGGTGCGGCAGGCTT
30
604
|
|
1132646
N/A
N/A
4875
4894
CCCTGGGCAGCCAAGGCTGG
53
605
|
|
1132674
N/A
N/A
5395
5414
CTCGGCTCCTCCTTCCCCCC
45
606
|
|
1132702
N/A
N/A
5763
5782
AAGCTGCGCTGGGAGACGGA
45
607
|
|
1132730
N/A
N/A
6170
6189
CGCAACAGAGCTAACCCGGG
45
608
|
|
1132758
N/A
N/A
6385
6404
ACCAAAGTCGCGGGCTTCTT
43
609
|
|
1132786
N/A
N/A
6493
6512
GGATCCCAGCAGTGAATCCC
59
610
|
|
1132814
N/A
N/A
6633
6652
ACCACCCATCCAGAGTCGCA
44
611
|
|
1132842
N/A
N/A
6724
6743
GCCATTAATTCAATTTCATA
45
612
|
|
1132870
N/A
N/A
6865
6884
CCGCCCCTGCGAACACAGAG
44
613
|
|
TABLE 40
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 2000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130868
213
232
3564
3583
ACATTTGTGGTACAGCTGCC
84
1756
|
|
1130987
443
462
N/A
N/A
GGCTCAAAGCACTTCTCTTT
93
2135
|
|
1131593
1780
1799
7168
7187
TGATGCCTTGCAGGGTGAGC
37
1326
|
|
1131694
1919
1938
7307
7326
CTGCGGAATCACCAAGGAGG
57
571
|
|
1131701
1928
1947
7316
7335
ACTCTCTCACTGCGGAATCA
42
1179
|
|
1131746
2005
2024
7393
7412
GAGTTCCTGCGCCATCCTGG
40
159
|
|
1131754
2015
2034
7403
7422
GCACTTTATTGAGTTCCTGC
35
164
|
|
1131756
2017
2036
7405
7424
AAGCACTTTATTGAGTTCCT
11
1030
|
|
1131757
2018
2037
7406
7425
AAAGCACTTTATTGAGTTCC
36
165
|
|
1131763
2026
2045
7414
7433
GCATTTTCAAAGCACTTTAT
41
1562
|
|
1131891
N/A
N/A
513
532
CCGACTGTGTGCTCTTCAGC
88
659
|
|
1132188
N/A
N/A
2323
2342
ACTGTTTGCTAGTTCAATGT
106
1951
|
|
1132271
N/A
N/A
2706
2725
CACATTCCCAGTTAAGGTTC
93
1879
|
|
1132505
N/A
N/A
4024
4043
CAAGTTTCCAAGCTTGGTTT
104
525
|
|
1132651
N/A
N/A
4908
4927
AGCTTGCCTTGGTGTCTGAG
79
985
|
|
1132797
N/A
N/A
6542
6561
AGGGAACCAAGAGCAAGTTC
72
1369
|
|
1194299
N/A
N/A
2318
2337
TTGCTAGTTCAATGTTCACT
95
2351
|
|
1194300
N/A
N/A
2319
2338
TTTGCTAGTTCAATGTTCAC
67
2352
|
|
1194301
N/A
N/A
2321
2340
TGTTTGCTAGTTCAATGTTC
102
2353
|
|
1194302
N/A
N/A
2322
2341
CTGTTTGCTAGTTCAATGTT
97
2354
|
|
1194303
N/A
N/A
2324
2343
TACTGTTTGCTAGTTCAATG
95
2355
|
|
1194304
N/A
N/A
2325
2344
ATACTGTTTGCTAGTTCAAT
97
2356
|
|
1194305
N/A
N/A
2327
2346
TAATACTGTTTGCTAGTTCA
96
2357
|
|
1194306
N/A
N/A
2328
2347
CTAATACTGTTTGCTAGTTC
93
2358
|
|
1194307
N/A
N/A
2701
2720
TCCCAGTTAAGGTTCAACAA
99
2359
|
|
1194308
N/A
N/A
2702
2721
TTCCCAGTTAAGGTTCAACA
96
2360
|
|
1194309
N/A
N/A
2704
2723
CATTCCCAGTTAAGGTTCAA
79
2361
|
|
1194310
N/A
N/A
2705
2724
ACATTCCCAGTTAAGGTTCA
81
2362
|
|
1194311
N/A
N/A
2707
2726
GCACATTCCCAGTTAAGGTT
100
2363
|
|
1194312
N/A
N/A
2708
2727
TGCACATTCCCAGTTAAGGT
95
2364
|
|
1194313
N/A
N/A
2710
2729
TTTGCACATTCCCAGTTAAG
92
2365
|
|
1194314
N/A
N/A
2711
2730
ATTTGCACATTCCCAGTTAA
105
2366
|
|
1194315
N/A
N/A
4019
4038
TTCCAAGCTTGGTTTACCCA
113
2367
|
|
1194316
N/A
N/A
4020
4039
TTTCCAAGCTTGGTTTACCC
109
2368
|
|
1194317
N/A
N/A
4022
4041
AGTTTCCAAGCTTGGTTTAC
87
2369
|
|
1194318
N/A
N/A
4023
4042
AAGTTTCCAAGCTTGGTTTA
84
2370
|
|
1194319
N/A
N/A
4025
4044
CCAAGTTTCCAAGCTTGGTT
102
2371
|
|
1194320
N/A
N/A
4026
4045
TCCAAGTTTCCAAGCTTGGT
81
2372
|
|
1194321
N/A
N/A
4028
4047
ACTCCAAGTTTCCAAGCTTG
95
2373
|
|
1194322
N/A
N/A
4029
4048
TACTCCAAGTTTCCAAGCTT
98
2374
|
|
1194323
N/A
N/A
4903
4922
GCCTTGGTGTCTGAGGAGAA
83
2375
|
|
1194324
N/A
N/A
4904
4923
TGCCTTGGTGTCTGAGGAGA
80
2376
|
|
1194325
N/A
N/A
4906
4925
CTTGCCTTGGTGTCTGAGGA
75
2377
|
|
1194326
N/A
N/A
4907
4926
GCTTGCCTTGGTGTCTGAGG
77
2378
|
|
1194327
N/A
N/A
4909
4928
CAGCTTGCCTTGGTGTCTGA
78
2379
|
|
1194328
N/A
N/A
4910
4929
GCAGCTTGCCTTGGTGTCTG
87
2380
|
|
1194329
683
702
4912
4931
TAGCAGCTTGCCTTGGTGTC
99
2381
|
|
1194330
684
703
4913
4932
ATAGCAGCTTGCCTTGGTGT
93
2382
|
|
1194331
N/A
N/A
6537
6556
ACCAAGAGCAAGTTCGATTC
60
2383
|
|
1194332
N/A
N/A
6538
6557
AACCAAGAGCAAGTTCGATT
56
2384
|
|
1194333
N/A
N/A
6540
6559
GGAACCAAGAGCAAGTTCGA
73
2385
|
|
1194334
N/A
N/A
6541
6560
GGGAACCAAGAGCAAGTTCG
69
2386
|
|
1194335
N/A
N/A
6543
6562
GAGGGAACCAAGAGCAAGTT
74
2387
|
|
1194336
N/A
N/A
6544
6563
AGAGGGAACCAAGAGCAAGT
69
2388
|
|
1194337
N/A
N/A
6545
6564
CAGAGGGAACCAAGAGCAAG
85
2389
|
|
1194338
N/A
N/A
6546
6565
CCAGAGGGAACCAAGAGCAA
68
2390
|
|
1194339
N/A
N/A
6547
6566
CCCAGAGGGAACCAAGAGCA
57
2391
|
|
1206452
143
162
510
529
ACTGTGTGCTCTTCAGCTTT
78
175
|
|
1206453
209
228
3560
3579
TTGTGGTACAGCTGCCGGTG
79
2392
|
|
1206454
1912
1931
7300
7319
ATCACCAAGGAGGGAAAGAT
53
2393
|
|
1206455
1920
1939
7308
7327
ACTGCGGAATCACCAAGGAG
39
2394
|
|
1206456
1925
1944
7313
7332
CTCTCACTGCGGAATCACCA
28
2395
|
|
1206457
1929
1948
7317
7336
CACTCTCTCACTGCGGAATC
17
2396
|
|
1206458
1931
1950
7319
7338
GCCACTCTCTCACTGCGGAA
25
2397
|
|
1206459
1934
1953
7322
7341
CCAGCCACTCTCTCACTGCG
42
2398
|
|
1206460
N/A
N/A
514
533
ACCGACTGTGTGCTCTTCAG
82
2399
|
|
1206461
N/A
N/A
515
534
TACCGACTGTGTGCTCTTCA
90
2400
|
|
1206462
N/A
N/A
517
536
CTTACCGACTGTGTGCTCTT
102
2401
|
|
1206463
N/A
N/A
518
537
ACTTACCGACTGTGTGCTCT
68
2402
|
|
1206464
N/A
N/A
2313
2332
AGTTCAATGTTCACTGTGAT
105
2403
|
|
1206465
N/A
N/A
2333
2352
GGAACCTAATACTGTTTGCT
92
2404
|
|
1206466
N/A
N/A
2696
2715
GTTAAGGTTCAACAAGGCGT
81
2405
|
|
1206467
N/A
N/A
2716
2735
GAAAAATTTGCACATTCCCA
92
2406
|
|
1206468
N/A
N/A
4014
4033
AGCTTGGTTTACCCACCTGC
95
2407
|
|
1206469
N/A
N/A
4034
4053
CTTGCTACTCCAAGTTTCCA
80
2408
|
|
1206470
N/A
N/A
4898
4917
GGTGTCTGAGGAGAAAGGGG
82
2409
|
|
1206471
N/A
N/A
6532
6551
GAGCAAGTTCGATTCTCCCT
67
2410
|
|
1206472
N/A
N/A
6552
6571
CGGCGCCCAGAGGGAACCAA
95
2411
|
|
TABLE 41
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 3
NO: 3
NO: 4
NO: 4
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1131768
N/A
N/A
2033
2052
TTTCTCAGCATTTTCAAAGC
35
2412
|
|
1131767
10843
10862
2031
2050
TCTCAGCATTTTCAAAGCAC
25
2413
|
|
1131770
N/A
N/A
2036
2055
TTTTTTCTCAGCATTTTCAA
54
2414
|
|
1131771
N/A
N/A
2037
2056
TTTTTTTCTCAGCATTTTCA
83
2415
|
|
1130767
3458
3477
34
53
TCATGGCATCCGTCCGTTGG
36
617
|
|
1130768
3459
3478
35
54
CTCATGGCATCCGTCCGTTG
90
619
|
|
1132897
10846
10865
N/A
N/A
CCTTCTCAGCATTTTCAAAG
62
618
|
|
1130769
3460
3479
36
55
CCTCATGGCATCCGTCCGTT
22
540
|
|
1132898
10849
10868
N/A
N/A
TTTCCTTCTCAGCATTTTCA
61
616
|
|
TABLE 42
|
|
Reduction of FXII RNA by 5-10-5 MOE gapmers with mixed PO/PS internucleoside linkages
|
(Huh7, electroporation, 5000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 3
NO: 3
NO: 4
NO: 4
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1130744
3428
3447
4
23
TATCCAGGAGTCCAGATCAA
61
2416
|
|
1130772
3463
3482
39
58
AGCCCTCATGGCATCCGTCC
35
2417
|
|
1130755
3443
3462
19
38
GTTGGTCCAGCTGCCTATCC
64
2418
|
|
1130756
3444
3463
20
39
CGTTGGTCCAGCTGCCTATC
50
2419
|
|
1130760
3450
3469
26
45
TCCGTCCGTTGGTCCAGCTG
37
2420
|
|
1130765
3456
3475
32
51
ATGGCATCCGTCCGTTGGTC
43
238
|
|
1130766
3457
3476
33
52
CATGGCATCCGTCCGTTGGT
50
314
|
|
Example 3: Effects of 3-10-3 cET Gapmers on Human FXII RNA In Vitro, Single Dose
Modified oligonucleotides complementary to an FXII nucleic acid were synthesized and tested for their effect on FXII RNA levels in vitro. The modified oligonucleotides were tested in a series of experiments using similar culture conditions. The results for each separate experiment are presented in separate tables below.
The modified oligonucleotides in the tables below are 3-10-3 cEt gapmers (i.e., they have a central region of ten 2′-deoxynucleosides flanked on each side by wings, each comprising three cEt modified nucleosides). The internucleoside linkages throughout each modified oligonucleotide are phosphorothioate (P═S) linkages. All cytosine nucleobases throughout each modified oligonucleotide are 5-methylcytosines.
“Start site” indicates the 5′-most nucleoside of the target sequence to which the modified oligonucleotide is complementary. “Stop site” indicates the 3′-most nucleoside of the target sequence to which the modified oligonucleotide is complementary. As shown in the tables below, the modified oligonucleotides are 100% complementary to one or more of human FXII target sequences including the human FXII mRNA sequence designated herein as SEQ ID NO: 1 (ENSEMBL ID ENST00000253496.3 from ENSEMBL version 99: January 2020), the human FXII genomic sequence, designated herein as SEQ ID NO: 2 (ENSEMBL ID ENSG00000131187.9 from ENSEML version 99: January 2020), the human FXII genomic sequence, designated herein as SEQ ID No. 3 (the complement of GENBANK Accession No. NC_000005.10 truncated from truncated from nucleotides 177399001 to 177413000), and the human FXII mRNA sequence designated herein as SEQ ID No: 4 (GENBANK Accession No. NM 000505.3) ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular target sequence with 100% complementarity.
Cultured Huh7 cells, at a density of 20,000 cells per well, were transfected using electroporation with either 2000 nM or 3000 nM modified oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and FXII RNA levels were measured by quantitative real-time RTPCR. Human primer probe set RTS40528 (described herein above) was used to measure RNA levels. FXII RNA levels were normalized to total RNA content, as measured by RIBOGREEN®. Reduction of FXII RNA is presented in the tables below as percent FXII RNA relative to the amount in untreated control cells (% UTC). The symbol “†” indicates that the modified oligonucleotide is complementary to the target transcript within the amplicon region of the primer probe set. In such instances, additional assays using alternative primer probes must be performed to accurately assess the potency and efficacy of such modified oligonucleotides.
TABLE 43
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128712
3
18
3
18
AGGAGTCCAGATCAAT
50
2421
|
|
1128732
80
95
80
95
ACTCCAAGCTCACCAG
37
2422
|
|
1128752
125
140
492
507
ACTTATGCTCCTTGGG
39
2423
|
|
1128772
156
171
N/A
N/A
GTGAGAACGACTGTGT
39
2424
|
|
1128792
200
215
3551
3566
GCCGGTGGTACTGGAA
57
2425
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
12
2426
|
|
1128813
231
246
3582
3597
CGGCCCTTGTGGGTAC
79
2427
|
|
1128833
294
309
3801
3816
TATCCCCATCGCTGGT
51
2428
|
|
1128853
409
424
4217
4232
GAGGTGTTGTGGACAG
45
2429
|
|
1128873
482
497
4427
4442
ATATCTCATTCTTGTG
22
2430
|
|
1128893
545
560
4490
4505
GGCAGTGGGCATCAGG
40
2431
|
|
1128913
598
613
4682
4697
GCGACCCCCATGGAGG
39
2432
|
|
1128933
634
649
4718
4733
GCAGTGGCACAGGCGG
36
2433
|
|
1128953
664
679
4748
4763
GTCGCAGAAGGCTCCG
64
2434
|
|
1128973
713
728
4942
4957
CGCGGTAGCTGAGCCC
68
2435
|
|
1128993
759
774
4988
5003
GCCCACGGCTGACAGG
62
2436
|
|
1129013
807
822
5036
5051
CAGTTCCGCGCTTGCT
60
2437
|
|
1129033
863
878
5177
5192
ACCACGGGCGGATGTC
54
2438
|
|
1129053
892
907
5206
5221
GCTCAGCCGGTCGCGG
51
2439
|
|
1129073
982
997
5296
5311
CATGAGTGGGACATGA
45
2440
|
|
1129093
1043
1058
5357
5372
TCTGGGACTGAGGCGG
33
2441
|
|
1129113
1089
1104
5508
5523
GTCAGGGAAGGCGGCT
38
2442
|
|
1129133
1129
1144
5548
5563
CTTGCGGAGCCGCTGC
37
2443
|
|
1129153
1152
1167
5571
5586
ACGCGGGTCATCGAAG
53
2444
|
|
1129173
1209
1224
5628
5643
CAGTACAGCGCGGCGA
35
2445
|
|
1129193
1240
1255
5659
5674
GATGAGGCTGCCGGCG
81
2446
|
|
1129213
1319
1334
5979
5994
GGCCGAGCACCACCGT
44
2447
|
|
1129233
1384
1399
6044
6059
GTGCAAGCGGTAGGAG
76
2448
|
|
1129253
1438
1453
6181
6196
CTGAAGGCGCAACAGA
61
2449
|
|
1129273
1463
1478
6206
6221
GCGCGCAGCTGCCGTC
39
2450
|
|
1129293
1497
1512
6240
6255
GGCAGGCACACCGGCT
93
2451
|
|
1129313
1558
1573
6301
6316
CTGGTGGCCCCAGCCG
96
2452
|
|
1129333
1580
1595
6878
6893
CATATTCCTCCGCCCC
78
2453
|
|
1129353
1619
1634
6917
6932
GGGAGAGGAACGGTAC
126
2454
|
|
1129373
1693
1708
6991
7006
CTCGAGGAACCCTGCG
27†
2455
|
|
1129393
1748
1763
7136
7151
GGTCCTCACACACCAG
44†
2456
|
|
1129413
1798
1813
7186
7201
CGATCCCCAGCTGATG
27
2457
|
|
1129433
1839
1854
7227
7242
ACATCGGTGTAGACGC
28
2458
|
|
1129453
1866
1881
7254
7269
TCCCGGATCCAGGCCA
41
2459
|
|
1129473
1898
1913
7286
7301
ATGAGTCCCTGAGCAA
26
2460
|
|
1129493
1931
1946
7319
7334
CTCTCTCACTGCGGAA
15
2461
|
|
1129513
1990
2005
7378
7393
GCGCGGAGCTGGCCGC
46
2462
|
|
1129533
2011
2026
7399
7414
TTGAGTTCCTGCGCCA
23
2463
|
|
1129553
N/A
N/A
97
112
ACTCACCGAAAGTGTT
78
2464
|
|
1129573
N/A
N/A
197
212
CCTAGTCACCTGGACC
63
2465
|
|
1129593
N/A
N/A
328
343
CCTCCCCCCGTTGTCT
49
2466
|
|
1129613
N/A
N/A
434
449
GAACTGACTATAAGTC
61
2467
|
|
1129633
N/A
N/A
606
621
GCACTAGACTAGACTG
54
2468
|
|
1129653
N/A
N/A
652
667
CACGATCACTCTAGTG
51
2469
|
|
1129673
N/A
N/A
908
923
GAGGCATTCAGATGCC
61
2470
|
|
1129693
N/A
N/A
1055
1070
GAATACCTCACATGGG
86
2471
|
|
1129713
N/A
N/A
1557
1572
GGGTAAGGTCCATCTG
61
2472
|
|
1129733
N/A
N/A
1709
1724
CAAGGATAGGGCACCA
50
2473
|
|
1129753
N/A
N/A
2146
2161
AGGCAAAGTGCACTTG
55
2474
|
|
1129773
N/A
N/A
2450
2465
GCATTGATGAATCAGC
47
2475
|
|
1129793
N/A
N/A
2527
2542
GCACAAGAAGGCTACC
54
2476
|
|
1129813
N/A
N/A
2852
2867
AATAGTACAGTTGATC
54
2477
|
|
1129833
N/A
N/A
3048
3063
CAGGAGTCAGATAGCT
64
2478
|
|
1129853
N/A
N/A
3652
3667
GCAGGGTACATGTCTC
68
2479
|
|
1129873
N/A
N/A
4015
4030
TTGGTTTACCCACCTG
73
2480
|
|
1129893
N/A
N/A
4316
4331
ATCCAGCAACCTATTC
67
2481
|
|
1129913
N/A
N/A
4360
4375
ATAGTGGTCTCAGGAG
64
2482
|
|
1129933
N/A
N/A
4619
4634
CTGGGCCTAAAGACCC
62
2483
|
|
1129953
N/A
N/A
5070
5085
GGCGCACCGGCAGAAG
52
2484
|
|
1129973
N/A
N/A
5418
5433
CTAGCTCGCCCGGCGC
76
2485
|
|
1129993
N/A
N/A
5450
5465
GGACGGAGAGCCCGCG
74
2486
|
|
1130013
N/A
N/A
5755
5770
GAGACGGAGGAGCCGC
54
2487
|
|
1130033
N/A
N/A
5869
5884
ACCCCAATCCCGTGTT
61
2488
|
|
1130053
N/A
N/A
6170
6185
ACAGAGCTAACCCGGG
96
2489
|
|
1130073
N/A
N/A
6330
6345
TGCCCCTAGCAGTTGT
65
2490
|
|
1130093
N/A
N/A
6389
6404
ACCAAAGTCGCGGGCT
137
2491
|
|
1130113
N/A
N/A
6413
6428
CATTCTGTAGGCACCC
83
2492
|
|
1130133
N/A
N/A
6538
6553
AAGAGCAAGTTCGATT
51
2493
|
|
1130153
N/A
N/A
6624
6639
AGTCGCAGAACCTGGC
49
2494
|
|
1130173
N/A
N/A
6676
6691
GATTTGAATGGGCGGA
55
2495
|
|
1130193
N/A
N/A
6812
6827
CGGACGATGGACAAAG
64
2496
|
|
1130213
N/A
N/A
6876
6891
TATTCCTCCGCCCCTG
131
2497
|
|
1130233
N/A
N/A
7078
7093
CGCTGTGGACCTGAGA
89†
2498
|
|
TABLE 44
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128713
4
19
4
19
CAGGAGTCCAGATCAA
35
2499
|
|
1128733
81
96
81
96
GACTCCAAGCTCACCA
38
2500
|
|
1128753
126
141
493
508
TACTTATGCTCCTTGG
35
2501
|
|
1128773
157
172
N/A
N/A
AGTGAGAACGACTGTG
31
2502
|
|
1128793
201
216
3552
3567
TGCCGGTGGTACTGGA
41
2503
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
18
2426
|
|
1128814
233
248
3584
3599
GCCGGCCCTTGTGGGT
55
2504
|
|
1128834
295
310
3802
3817
GTATCCCCATCGCTGG
35
2505
|
|
1128854
411
426
4219
4234
GTGAGGTGTTGTGGAC
38
2506
|
|
1128874
484
499
4429
4444
CCATATCTCATTCTTG
24
2507
|
|
1128894
551
566
4496
4511
GCCGCTGGCAGTGGGC
51
2508
|
|
1128914
599
614
4683
4698
AGCGACCCCCATGGAG
37
2509
|
|
1128934
639
654
4723
4738
ACCGGGCAGTGGCACA
46
2510
|
|
1128954
665
680
4749
4764
CGTCGCAGAAGGCTCC
45
2511
|
|
1128974
714
729
4943
4958
CCGCGGTAGCTGAGCC
90
2512
|
|
1128994
761
776
4990
5005
AGGCCCACGGCTGACA
44
2513
|
|
1129014
808
823
5037
5052
CCAGTTCCGCGCTTGC
33
2514
|
|
1129034
864
879
5178
5193
CACCACGGGCGGATGT
56
2515
|
|
1129054
896
911
5210
5225
CCCAGCTCAGCCGGTC
57
2516
|
|
1129074
989
1004
5303
5318
GCGCGGGCATGAGTGG
37
2517
|
|
1129094
1044
1059
5358
5373
GTCTGGGACTGAGGCG
26
2518
|
|
1129114
1090
1105
5509
5524
GGTCAGGGAAGGCGGC
21
2519
|
|
1129134
1130
1145
5549
5564
TCTTGCGGAGCCGCTG
38
2520
|
|
1129154
1153
1168
5572
5587
GACGCGGGTCATCGAA
47
2521
|
|
1129174
1210
1225
5629
5644
CCAGTACAGCGCGGCG
27
2522
|
|
1129194
1241
1256
5660
5675
CGATGAGGCTGCCGGC
53
2523
|
|
1129214
1320
1335
5980
5995
TGGCCGAGCACCACCG
38
2524
|
|
1129234
1385
1400
6045
6060
CGTGCAAGCGGTAGGA
38
2525
|
|
1129254
1439
1454
6182
6197
CCTGAAGGCGCAACAG
37
2526
|
|
1129274
1464
1479
6207
6222
AGCGCGCAGCTGCCGT
30
2527
|
|
1129294
1498
1513
6241
6256
TGGCAGGCACACCGGC
20
2528
|
|
1129314
1559
1574
6302
6317
ACTGGTGGCCCCAGCC
73
2529
|
|
1129334
1581
1596
6879
6894
GCATATTCCTCCGCCC
83
2530
|
|
1129354
1620
1635
6918
6933
AGGGAGAGGAACGGTA
69
2531
|
|
1129374
1694
1709
6992
7007
CCTCGAGGAACCCTGC
18†
2532
|
|
1129394
1749
1764
7137
7152
TGGTCCTCACACACCA
41†
2533
|
|
1129414
1800
1815
7188
7203
CCCGATCCCCAGCTGA
47
2534
|
|
1129434
1840
1855
7228
7243
CACATCGGTGTAGACG
34
2535
|
|
1129454
1867
1882
7255
7270
CTCCCGGATCCAGGCC
47
2536
|
|
1129474
1899
1914
7287
7302
GATGAGTCCCTGAGCA
27
2537
|
|
1129494
1946
1961
7334
7349
TCCATGCCCCAGCCAC
25
2538
|
|
1129514
1991
2006
7379
7394
GGCGCGGAGCTGGCCG
77
2539
|
|
1129534
2012
2027
7400
7415
ATTGAGTTCCTGCGCC
15
2540
|
|
1129554
N/A
N/A
99
114
GCACTCACCGAAAGTG
78
2541
|
|
1129574
N/A
N/A
198
213
TCCTAGTCACCTGGAC
53
2542
|
|
1129594
N/A
N/A
329
344
ACCTCCCCCCGTTGTC
62
2543
|
|
1129614
N/A
N/A
435
450
GGAACTGACTATAAGT
53
2544
|
|
1129634
N/A
N/A
607
622
GGCACTAGACTAGACT
64
2545
|
|
1129654
N/A
N/A
653
668
TCACGATCACTCTAGT
41
2546
|
|
1129674
N/A
N/A
953
968
CTCGCCCAGAGTCACC
72
2547
|
|
1129694
N/A
N/A
1056
1071
TGAATACCTCACATGG
48
2548
|
|
1129714
N/A
N/A
1558
1573
AGGGTAAGGTCCATCT
35
2549
|
|
1129734
N/A
N/A
1710
1725
GCAAGGATAGGGCACC
57
2550
|
|
1129754
N/A
N/A
2250
2265
TTCAGAGGAATTGTCG
52
2551
|
|
1129774
N/A
N/A
2452
2467
TAGCATTGATGAATCA
72
2552
|
|
1129794
N/A
N/A
2540
2555
GTGTAGTTCCTAAGCA
57
2553
|
|
1129814
N/A
N/A
2862
2877
CGGCAGTAATAATAGT
77
2554
|
|
1129834
N/A
N/A
3373
3388
TCTAAAAGTTGGGTTC
51
2555
|
|
1129854
N/A
N/A
3704
3719
GCCCTATCACAGTCCC
53
2556
|
|
1129874
N/A
N/A
4017
4032
GCTTGGTTTACCCACC
70
2557
|
|
1129894
N/A
N/A
4318
4333
GTATCCAGCAACCTAT
48
2558
|
|
1129914
N/A
N/A
4361
4376
GATAGTGGTCTCAGGA
58
2559
|
|
1129934
N/A
N/A
4620
4635
CCTGGGCCTAAAGACC
85
2560
|
|
1129954
N/A
N/A
5071
5086
CGGCGCACCGGCAGAA
42
2561
|
|
1129974
N/A
N/A
5419
5434
TCTAGCTCGCCCGGCG
61
2562
|
|
1129994
N/A
N/A
5451
5466
AGGACGGAGAGCCCGC
49
2563
|
|
1130014
N/A
N/A
5762
5777
GCGCTGGGAGACGGAG
101
2564
|
|
1130034
N/A
N/A
5870
5885
AACCCCAATCCCGTGT
54
2565
|
|
1130054
N/A
N/A
6171
6186
AACAGAGCTAACCCGG
62
2566
|
|
1130074
N/A
N/A
6331
6346
CTGCCCCTAGCAGTTG
118
2567
|
|
1130094
N/A
N/A
6390
6405
TACCAAAGTCGCGGGC
46
2568
|
|
1130114
N/A
N/A
6420
6435
CGCCACCCATTCTGTA
54
2569
|
|
1130134
N/A
N/A
6539
6554
CAAGAGCAAGTTCGAT
69
2570
|
|
1130154
N/A
N/A
6626
6641
AGAGTCGCAGAACCTG
51
2571
|
|
1130174
N/A
N/A
6677
6692
GGATTTGAATGGGCGG
48
2572
|
|
1130194
N/A
N/A
6813
6828
CCGGACGATGGACAAA
56
2573
|
|
1130214
N/A
N/A
6877
6892
ATATTCCTCCGCCCCT
78
2574
|
|
1130234
N/A
N/A
7091
7106
ACACGCAGCTCAGCGC
56†
2575
|
|
TABLE 45
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128714
6
21
6
21
TCCAGGAGTCCAGATC
85
2576
|
|
1128734
82
97
82
97
TGACTCCAAGCTCACC
30
2577
|
|
1128754
127
142
494
509
GTACTTATGCTCCTTG
16
2578
|
|
1128774
158
173
N/A
N/A
CAGTGAGAACGACTGT
36
2579
|
|
1128794
202
217
3553
3568
CTGCCGGTGGTACTGG
22
2580
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
12
2426
|
|
1128815
235
250
3586
3601
TGGCCGGCCCTTGTGG
57
2581
|
|
1128835
297
312
3804
3819
CAGTATCCCCATCGCT
16
2582
|
|
1128855
412
427
4220
4235
AGTGAGGTGTTGTGGA
38
2583
|
|
1128875
487
502
4432
4447
ATACCATATCTCATTC
18
2584
|
|
1128895
552
567
4497
4512
AGCCGCTGGCAGTGGG
49
2585
|
|
1128915
600
615
4684
4699
CAGCGACCCCCATGGA
30
2586
|
|
1128935
640
655
4724
4739
CACCGGGCAGTGGCAC
32
2587
|
|
1128955
679
694
N/A
N/A
TGCCTTGGTGTCCACG
31
2588
|
|
1128975
715
730
4944
4959
GCCGCGGTAGCTGAGC
52
2589
|
|
1128995
762
777
4991
5006
GAGGCCCACGGCTGAC
31
2590
|
|
1129015
809
824
5038
5053
CCCAGTTCCGCGCTTG
18
2591
|
|
1129035
866
881
5180
5195
AGCACCACGGGCGGAT
26
2592
|
|
1129055
897
912
5211
5226
TCCCAGCTCAGCCGGT
27
2593
|
|
1129075
990
1005
5304
5319
TGCGCGGGCATGAGTG
24
2594
|
|
1129095
1060
1075
N/A
N/A
CGGCAAGGCTCCCGGG
35
2595
|
|
1129115
1099
1114
5518
5533
GCCGTTCCTGGTCAGG
25
2596
|
|
1129135
1131
1146
5550
5565
CTCTTGCGGAGCCGCT
22
2597
|
|
1129155
1154
1169
5573
5588
CGACGCGGGTCATCGA
58
2598
|
|
1129175
1211
1226
5630
5645
CCCAGTACAGCGCGGC
22
2599
|
|
1129195
1242
1257
5661
5676
GCGATGAGGCTGCCGG
30
2600
|
|
1129215
1321
1336
5981
5996
CTGGCCGAGCACCACC
23
2601
|
|
1129235
1399
1414
6059
6074
GGGCGAGAAGGCCTCG
60
2602
|
|
1129255
1440
1455
6183
6198
TCCTGAAGGCGCAACA
22
2603
|
|
1129275
1465
1480
6208
6223
GAGCGCGCAGCTGCCG
18
2604
|
|
1129295
1499
1514
6242
6257
TTGGCAGGCACACCGG
20
2605
|
|
1129315
1560
1575
6303
6318
AACTGGTGGCCCCAGC
38
2606
|
|
1129335
1582
1597
6880
6895
GGCATATTCCTCCGCC
42
2607
|
|
1129355
1621
1636
6919
6934
CAGGGAGAGGAACGGT
43
2608
|
|
1129375
1695
1710
6993
7008
CCCTCGAGGAACCCTG
9†
2609
|
|
1129395
1750
1765
7138
7153
TTGGTCCTCACACACC
24†
2610
|
|
1129415
1801
1816
7189
7204
GCCCGATCCCCAGCTG
33
2611
|
|
1129435
1841
1856
7229
7244
CCACATCGGTGTAGAC
20
2612
|
|
1129455
1868
1883
7256
7271
GCTCCCGGATCCAGGC
34
2613
|
|
1129475
1900
1915
7288
7303
AGATGAGTCCCTGAGC
10
2614
|
|
1129495
1947
1962
7335
7350
TTCCATGCCCCAGCCA
20
2615
|
|
1129515
1992
2007
7380
7395
TGGCGCGGAGCTGGCC
49
2616
|
|
1129535
2013
2028
7401
7416
TATTGAGTTCCTGCGC
8
2617
|
|
1129555
N/A
N/A
100
115
AGCACTCACCGAAAGT
105
2618
|
|
1129575
N/A
N/A
199
214
CTCCTAGTCACCTGGA
80
2619
|
|
1129595
N/A
N/A
332
347
TCTACCTCCCCCCGTT
117
2620
|
|
1129615
N/A
N/A
436
451
GGGAACTGACTATAAG
153
2621
|
|
1129635
N/A
N/A
608
623
AGGCACTAGACTAGAC
62
2622
|
|
1129655
N/A
N/A
654
669
CTCACGATCACTCTAG
56
2623
|
|
1129675
N/A
N/A
955
970
CCCTCGCCCAGAGTCA
58
2624
|
|
1129695
N/A
N/A
1059
1074
GGCTGAATACCTCACA
36
2625
|
|
1129715
N/A
N/A
1559
1574
CAGGGTAAGGTCCATC
33
2626
|
|
1129735
N/A
N/A
1711
1726
GGCAAGGATAGGGCAC
50
2627
|
|
1129755
N/A
N/A
2279
2294
CGCGAATAATGAGAAT
64
2628
|
|
1129775
N/A
N/A
2457
2472
CAGCTTAGCATTGATG
52
2629
|
|
1129795
N/A
N/A
2543
2558
GCTGTGTAGTTCCTAA
56
2630
|
|
1129815
N/A
N/A
2863
2878
ACGGCAGTAATAATAG
49
2631
|
|
1129835
N/A
N/A
3375
3390
GCTCTAAAAGTTGGGT
60
2632
|
|
1129855
N/A
N/A
3764
3779
TAGCACACCTGTAGAA
83
2633
|
|
1129875
N/A
N/A
4018
4033
AGCTTGGTTTACCCAC
42
2634
|
|
1129895
N/A
N/A
4320
4335
GAGTATCCAGCAACCT
49
2635
|
|
1129915
N/A
N/A
4362
4377
GGATAGTGGTCTCAGG
65
2636
|
|
1129935
N/A
N/A
4622
4637
CCCCTGGGCCTAAAGA
72
2637
|
|
1129955
N/A
N/A
5072
5087
GCGGCGCACCGGCAGA
55
2638
|
|
1129975
N/A
N/A
5420
5435
ATCTAGCTCGCCCGGC
37
2639
|
|
1129995
N/A
N/A
5452
5467
GAGGACGGAGAGCCCG
88
2640
|
|
1130015
N/A
N/A
5763
5778
TGCGCTGGGAGACGGA
73
2641
|
|
1130035
N/A
N/A
5871
5886
GAACCCCAATCCCGTG
49
2642
|
|
1130055
N/A
N/A
6172
6187
CAACAGAGCTAACCCG
59
2643
|
|
1130075
N/A
N/A
6355
6370
GATCAAAGGTCTCCTC
41
2644
|
|
1130095
N/A
N/A
6391
6406
ATACCAAAGTCGCGGG
80
2645
|
|
1130115
N/A
N/A
6434
6449
ACCCATCAGGTCAGCG
59
2646
|
|
1130135
N/A
N/A
6540
6555
CCAAGAGCAAGTTCGA
47
2647
|
|
1130155
N/A
N/A
6627
6642
CAGAGTCGCAGAACCT
48
2648
|
|
1130175
N/A
N/A
6678
6693
AGGATTTGAATGGGCG
38
2649
|
|
1130195
N/A
N/A
6814
6829
CCCGGACGATGGACAA
68
2650
|
|
1130215
N/A
N/A
7015
7030
CACCTGGCACGCATCG
24†
2651
|
|
1130235
N/A
N/A
7093
7108
AAACACGCAGCTCAGC
81†
2652
|
|
TABLE 46
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128715
9
24
9
24
CTATCCAGGAGTCCAG
70
2653
|
|
1128735
87
102
87
102
AGTGTTGACTCCAAGC
22
2654
|
|
1128755
128
143
495
510
TGTACTTATGCTCCTT
23
2655
|
|
1128775
159
174
N/A
N/A
ACAGTGAGAACGACTG
48
2656
|
|
1128795
210
225
3561
3576
TGGTACAGCTGCCGGT
17
2657
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
8
2426
|
|
1128816
236
251
3587
3602
CTGGCCGGCCCTTGTG
35
2658
|
|
1128836
298
313
3805
3820
ACAGTATCCCCATCGC
16
2659
|
|
1128856
413
428
4221
4236
CAGTGAGGTGTTGTGG
34
2660
|
|
1128876
489
504
4434
4449
CTATACCATATCTCAT
20
2661
|
|
1128896
556
571
4501
4516
GGCCAGCCGCTGGCAG
74
2662
|
|
1128916
601
616
4685
4700
GCAGCGACCCCCATGG
19
2663
|
|
1128936
641
656
4725
4740
CCACCGGGCAGTGGCA
28
2664
|
|
1128956
683
698
4912
4927
AGCTTGCCTTGGTGTC
22
2665
|
|
1128976
716
731
4945
4960
GGCCGCGGTAGCTGAG
62
2666
|
|
1128996
765
780
4994
5009
TCCGAGGCCCACGGCT
30
2667
|
|
1129016
810
825
5039
5054
CCCCAGTTCCGCGCTT
36
2668
|
|
1129036
867
882
5181
5196
AAGCACCACGGGCGGA
25
2669
|
|
1129056
905
920
5219
5234
CGCAGTACTCCCAGCT
20
2670
|
|
1129076
991
1006
5305
5320
CTGCGCGGGCATGAGT
23
2671
|
|
1129096
1061
1076
N/A
N/A
CCGGCAAGGCTCCCGG
66
2672
|
|
1129116
1102
1117
5521
5536
TGGGCCGTTCCTGGTC
31
2673
|
|
1129136
1132
1147
5551
5566
ACTCTTGCGGAGCCGC
16
2674
|
|
1129156
1156
1171
5575
5590
AACGACGCGGGTCATC
54
2675
|
|
1129176
1212
1227
5631
5646
CCCCAGTACAGCGCGG
41
2676
|
|
1129196
1243
1258
5662
5677
GGCGATGAGGCTGCCG
79
2677
|
|
1129216
1323
1338
5983
5998
TCCTGGCCGAGCACCA
32
2678
|
|
1129236
1400
1415
6060
6075
CGGGCGAGAAGGCCTC
46
2679
|
|
1129256
1441
1456
6184
6199
CTCCTGAAGGCGCAAC
34
2680
|
|
1129276
1467
1482
6210
6225
AGGAGCGCGCAGCTGC
18
2681
|
|
1129296
1502
1517
6245
6260
CGCTTGGCAGGCACAC
29
2682
|
|
1129316
1561
1576
6304
6319
GAACTGGTGGCCCCAG
32
2683
|
|
1129336
1583
1598
6881
6896
TGGCATATTCCTCCGC
37
2684
|
|
1129356
1657
1672
6955
6970
GGAGGATCCGTGCACG
3†
2685
|
|
1129376
1696
1711
6994
7009
GCCCTCGAGGAACCCT
11†
2686
|
|
1129396
1751
1766
7139
7154
CTTGGTCCTCACACAC
9†
2687
|
|
1129416
1802
1817
7190
7205
AGCCCGATCCCCAGCT
36
2688
|
|
1129436
1842
1857
7230
7245
GCCACATCGGTGTAGA
23
2689
|
|
1129456
1876
1891
7264
7279
AACGGTGTGCTCCCGG
17
2690
|
|
1129476
1901
1916
7289
7304
AAGATGAGTCCCTGAG
7
2691
|
|
1129496
1948
1963
7336
7351
CTTCCATGCCCCAGCC
16
2692
|
|
1129516
1993
2008
7381
7396
CTGGCGCGGAGCTGGC
12
2693
|
|
1129536
2014
2029
7402
7417
TTATTGAGTTCCTGCG
10
2694
|
|
1129556
N/A
N/A
101
116
CAGCACTCACCGAAAG
55
2695
|
|
1129576
N/A
N/A
204
219
TAGGCCTCCTAGTCAC
57
2696
|
|
1129596
N/A
N/A
333
348
TTCTACCTCCCCCCGT
92
2697
|
|
1129616
N/A
N/A
439
454
GCAGGGAACTGACTAT
143
2698
|
|
1129636
N/A
N/A
610
625
GTAGGCACTAGACTAG
52
2699
|
|
1129656
N/A
N/A
655
670
GCTCACGATCACTCTA
33
2700
|
|
1129676
N/A
N/A
957
972
ATCCCTCGCCCAGAGT
66
2701
|
|
1129696
N/A
N/A
1061
1076
TGGGCTGAATACCTCA
52
2702
|
|
1129716
N/A
N/A
1560
1575
CCAGGGTAAGGTCCAT
45
2703
|
|
1129736
N/A
N/A
1713
1728
TAGGCAAGGATAGGGC
66
2704
|
|
1129756
N/A
N/A
2280
2295
TCGCGAATAATGAGAA
38
2705
|
|
1129776
N/A
N/A
2459
2474
ATCAGCTTAGCATTGA
55
2706
|
|
1129796
N/A
N/A
2628
2643
TCTACTTAGTGCAACG
30
2707
|
|
1129816
N/A
N/A
2910
2925
TCACTGTTAACCACTC
52
2708
|
|
1129836
N/A
N/A
3376
3391
TGCTCTAAAAGTTGGG
63
2709
|
|
1129856
N/A
N/A
3770
3785
GGGTGGTAGCACACCT
55
2710
|
|
1129876
N/A
N/A
4019
4034
AAGCTTGGTTTACCCA
49
2711
|
|
1129896
N/A
N/A
4321
4336
CGAGTATCCAGCAACC
83
2712
|
|
1129916
N/A
N/A
4363
4378
GGGATAGTGGTCTCAG
32
2713
|
|
1129936
N/A
N/A
4624
4639
CACCCCTGGGCCTAAA
75
2714
|
|
1129956
N/A
N/A
5073
5088
CGCGGCGCACCGGCAG
55
2715
|
|
1129976
N/A
N/A
5421
5436
AATCTAGCTCGCCCGG
39
2716
|
|
1129996
N/A
N/A
5453
5468
TGAGGACGGAGAGCCC
53
2717
|
|
1130016
N/A
N/A
5764
5779
CTGCGCTGGGAGACGG
64
2718
|
|
1130036
N/A
N/A
5872
5887
CGAACCCCAATCCCGT
57
2719
|
|
1130056
N/A
N/A
6174
6189
CGCAACAGAGCTAACC
43
2720
|
|
1130076
N/A
N/A
6360
6375
CCAGTGATCAAAGGTC
71
2721
|
|
1130096
N/A
N/A
6392
6407
GATACCAAAGTCGCGG
44
2722
|
|
1130116
N/A
N/A
6437
6452
ACAACCCATCAGGTCA
54
2723
|
|
1130136
N/A
N/A
6541
6556
ACCAAGAGCAAGTTCG
51
2724
|
|
1130156
N/A
N/A
6628
6643
CCAGAGTCGCAGAACC
47
2725
|
|
1130176
N/A
N/A
6752
6767
CAGCAAGCCCGAAGGG
60
2726
|
|
1130196
N/A
N/A
6815
6830
GCCCGGACGATGGACA
47
2727
|
|
1130216
N/A
N/A
7016
7031
TCACCTGGCACGCATC
32†
2728
|
|
1130236
N/A
N/A
7094
7109
GAAACACGCAGCTCAG
74†
2729
|
|
TABLE 47
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128716
11
26
11
26
GCCTATCCAGGAGTCC
73
2730
|
|
1128736
89
104
89
104
AAAGTGTTGACTCCAA
21
2731
|
|
1128756
129
144
496
511
TTGTACTTATGCTCCT
34
2732
|
|
1128776
160
175
N/A
N/A
GACAGTGAGAACGACT
14
2733
|
|
1128796
212
227
3563
3578
TGTGGTACAGCTGCCG
11
2734
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
7
2426
|
|
1128817
238
253
3589
3604
GCCTGGCCGGCCCTTG
46
2735
|
|
1128837
299
314
3806
3821
AACAGTATCCCCATCG
30
2736
|
|
1128857
414
429
4222
4237
CCAGTGAGGTGTTGTG
29
2737
|
|
1128877
492
507
4437
4452
GTTCTATACCATATCT
24
2738
|
|
1128897
572
587
N/A
N/A
TGCGGCAGGCCTGGCT
72
2739
|
|
1128917
602
617
4686
4701
GGCAGCGACCCCCATG
24
2740
|
|
1128937
642
657
4726
4741
CCCACCGGGCAGTGGC
50
2741
|
|
1128957
686
701
4915
4930
AGCAGCTTGCCTTGGT
19
2742
|
|
1128977
718
733
4947
4962
CAGGCCGCGGTAGCTG
70
2743
|
|
1128997
766
781
4995
5010
CTCCGAGGCCCACGGC
43
2744
|
|
1129017
811
826
5040
5055
TCCCCAGTTCCGCGCT
47
2745
|
|
1129037
869
884
5183
5198
CGAAGCACCACGGGCG
14
2746
|
|
1129057
906
921
5220
5235
TCGCAGTACTCCCAGC
15
2747
|
|
1129077
993
1008
5307
5322
GGCTGCGCGGGCATGA
26
2748
|
|
1129097
1062
1077
N/A
N/A
GCCGGCAAGGCTCCCG
34
2749
|
|
1129117
1104
1119
5523
5538
AGTGGGCCGTTCCTGG
23
2750
|
|
1129137
1133
1148
5552
5567
GACTCTTGCGGAGCCG
16
2751
|
|
1129157
1157
1172
5576
5591
CAACGACGCGGGTCAT
39
2752
|
|
1129177
1215
1230
5634
5649
TGGCCCCAGTACAGCG
60
2753
|
|
1129197
1244
1259
5663
5678
GGGCGATGAGGCTGCC
67
2754
|
|
1129217
1349
1364
6009
6024
ACGGCTCACAGCTGTG
67
2755
|
|
1129237
1401
1416
6061
6076
ACGGGCGAGAAGGCCT
51
2756
|
|
1129257
1442
1457
6185
6200
CCTCCTGAAGGCGCAA
33
2757
|
|
1129277
1468
1483
6211
6226
CAGGAGCGCGCAGCTG
29
2758
|
|
1129297
1503
1518
6246
6261
CCGCTTGGCAGGCACA
26
2759
|
|
1129317
1562
1577
6305
6320
CGAACTGGTGGCCCCA
35
2760
|
|
1129337
1584
1599
6882
6897
CTGGCATATTCCTCCG
47
2761
|
|
1129357
1658
1673
6956
6971
TGGAGGATCCGTGCAC
4†
2762
|
|
1129377
1697
1712
6995
7010
CGCCCTCGAGGAACCC
13†
2763
|
|
1129397
1752
1767
7140
7155
GCTTGGTCCTCACACA
6†
2764
|
|
1129417
1803
1818
7191
7206
CAGCCCGATCCCCAGC
24
2765
|
|
1129437
1843
1858
7231
7246
GGCCACATCGGTGTAG
45
2766
|
|
1129457
1877
1892
7265
7280
AAACGGTGTGCTCCCG
15
2767
|
|
1129477
1902
1917
7290
7305
AAAGATGAGTCCCTGA
9
2768
|
|
1129497
1949
1964
7337
7352
CCTTCCATGCCCCAGC
4
2769
|
|
1129517
1994
2009
7382
7397
CCTGGCGCGGAGCTGG
20
2770
|
|
1129537
2016
2031
7404
7419
CTTTATTGAGTTCCTG
4
2771
|
|
1129557
N/A
N/A
119
134
GGACAATCCTGGTTCC
55
2772
|
|
1129577
N/A
N/A
206
221
CATAGGCCTCCTAGTC
88
2773
|
|
1129597
N/A
N/A
334
349
TTTCTACCTCCCCCCG
77
2774
|
|
1129617
N/A
N/A
463
478
GGAATCTACAAGGGAG
76
2775
|
|
1129637
N/A
N/A
612
627
AGGTAGGCACTAGACT
67
2776
|
|
1129657
N/A
N/A
659
674
CACAGCTCACGATCAC
58
2777
|
|
1129677
N/A
N/A
958
973
AATCCCTCGCCCAGAG
71
2778
|
|
1129697
N/A
N/A
1125
1140
TCATCTAAAAGGTAGT
47
2779
|
|
1129717
N/A
N/A
1561
1576
ACCAGGGTAAGGTCCA
56
2780
|
|
1129737
N/A
N/A
1716
1731
GATTAGGCAAGGATAG
66
2781
|
|
1129757
N/A
N/A
2281
2296
ATCGCGAATAATGAGA
58
2782
|
|
1129777
N/A
N/A
2468
2483
TATAGTGCCATCAGCT
60
2783
|
|
1129797
N/A
N/A
2629
2644
GTCTACTTAGTGCAAC
64
2784
|
|
1129817
N/A
N/A
2912
2927
AGTCACTGTTAACCAC
51
2785
|
|
1129837
N/A
N/A
3401
3416
GTCGCTGTGCATTGAA
42
2786
|
|
1129857
N/A
N/A
3881
3896
GTAATGAGGCGGGAGG
62
2787
|
|
1129877
N/A
N/A
4035
4050
GCTACTCCAAGTTTCC
40
2788
|
|
1129897
N/A
N/A
4322
4337
CCGAGTATCCAGCAAC
53
2789
|
|
1129917
N/A
N/A
4365
4380
GAGGGATAGTGGTCTC
95
2790
|
|
1129937
N/A
N/A
4634
4649
CGCAGTGAGCCACCCC
71
2791
|
|
1129957
N/A
N/A
5074
5089
ACGCGGCGCACCGGCA
55
2792
|
|
1129977
N/A
N/A
5422
5437
GAATCTAGCTCGCCCG
78
2793
|
|
1129997
N/A
N/A
5482
5497
CGCCGGCAAGGCTGTG
77
2794
|
|
1130017
N/A
N/A
5766
5781
AGCTGCGCTGGGAGAC
79
2795
|
|
1130037
N/A
N/A
5873
5888
CCGAACCCCAATCCCG
97
2796
|
|
1130057
N/A
N/A
6175
6190
GCGCAACAGAGCTAAC
57
2797
|
|
1130077
N/A
N/A
6367
6382
GCCTAACCCAGTGATC
95
2798
|
|
1130097
N/A
N/A
6393
6408
CGATACCAAAGTCGCG
97
2799
|
|
1130117
N/A
N/A
6438
6453
CACAACCCATCAGGTC
57
2800
|
|
1130137
N/A
N/A
6567
6582
CCTTTGCAGCCCGGCG
69
2801
|
|
1130157
N/A
N/A
6629
6644
TCCAGAGTCGCAGAAC
61
2802
|
|
1130177
N/A
N/A
6753
6768
GCAGCAAGCCCGAAGG
65
2803
|
|
1130197
N/A
N/A
6816
6831
CGCCCGGACGATGGAC
59
2804
|
|
1130217
N/A
N/A
7017
7032
CTCACCTGGCACGCAT
64†
2805
|
|
1130237
N/A
N/A
7095
7110
GGAAACACGCAGCTCA
73†
2806
|
|
TABLE 48
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128717
12
27
12
27
TGCCTATCCAGGAGTC
97
2807
|
|
1128737
90
105
90
105
GAAAGTGTTGACTCCA
24
2808
|
|
1128757
131
146
498
513
CTTTGTACTTATGCTC
46
2809
|
|
1128777
161
176
N/A
N/A
TGACAGTGAGAACGAC
31
2810
|
|
1128797
213
228
3564
3579
TTGTGGTACAGCTGCC
15
2811
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
18
2426
|
|
1128818
243
258
3594
3609
TGAGGGCCTGGCCGGC
68
2812
|
|
1128838
300
315
3807
3822
AAACAGTATCCCCATC
57
2813
|
|
1128858
415
430
4223
4238
TCCAGTGAGGTGTTGT
35
2814
|
|
1128878
493
508
4438
4453
AGTTCTATACCATATC
15
2815
|
|
1128898
581
596
4665
4680
ACGGGTTGGTGCGGCA
44
2816
|
|
1128918
603
618
4687
4702
AGGCAGCGACCCCCAT
33
2817
|
|
1128938
643
658
4727
4742
GCCCACCGGGCAGTGG
83
2818
|
|
1128958
689
704
4918
4933
CATAGCAGCTTGCCTT
17
2819
|
|
1128978
720
735
4949
4964
GCCAGGCCGCGGTAGC
63
2820
|
|
1128998
767
782
4996
5011
CCTCCGAGGCCCACGG
35
2821
|
|
1129018
812
827
5041
5056
GTCCCCAGTTCCGCGC
66
2822
|
|
1129038
870
885
5184
5199
ACGAAGCACCACGGGC
30
2823
|
|
1129058
907
922
5221
5236
GTCGCAGTACTCCCAG
19
2824
|
|
1129078
995
1010
5309
5324
CCGGCTGCGCGGGCAT
33
2825
|
|
1129098
1063
1078
N/A
N/A
CGCCGGCAAGGCTCCC
42
2826
|
|
1129118
1106
1121
5525
5540
TCAGTGGGCCGTTCCT
30
2827
|
|
1129138
1134
1149
5553
5568
AGACTCTTGCGGAGCC
21
2828
|
|
1129158
1158
1173
5577
5592
CCAACGACGCGGGTCA
29
2829
|
|
1129178
1220
1235
5639
5654
AACTGTGGCCCCAGTA
42
2830
|
|
1129198
1261
1276
5680
5695
CGTCAGCACCCAGCAG
54
2831
|
|
1129218
1352
1367
6012
6027
GGCACGGCTCACAGCT
34
2832
|
|
1129238
1402
1417
6062
6077
GACGGGCGAGAAGGCC
42
2833
|
|
1129258
1443
1458
6186
6201
TCCTCCTGAAGGCGCA
38
2834
|
|
1129278
1469
1484
6212
6227
ACAGGAGCGCGCAGCT
37
2835
|
|
1129298
1504
1519
6247
6262
GCCGCTTGGCAGGCAC
77
2836
|
|
1129318
1563
1578
6306
6321
TCGAACTGGTGGCCCC
76
2837
|
|
1129338
1591
1606
6889
6904
CAGGAAGCTGGCATAT
59
2838
|
|
1129358
1659
1674
6957
6972
ATGGAGGATCCGTGCA
10†
2839
|
|
1129378
1698
1713
6996
7011
CCGCCCTCGAGGAACC
16†
2840
|
|
1129398
1753
1768
7141
7156
AGCTTGGTCCTCACAC
22†
2841
|
|
1129418
1806
1821
7194
7209
CCACAGCCCGATCCCC
36
2842
|
|
1129438
1844
1859
7232
7247
AGGCCACATCGGTGTA
34
2843
|
|
1129458
1878
1893
7266
7281
GAAACGGTGTGCTCCC
21
2844
|
|
1129478
1903
1918
7291
7306
GAAAGATGAGTCCCTG
15
2845
|
|
1129498
1950
1965
7338
7353
GCCTTCCATGCCCCAG
16
2846
|
|
1129518
1995
2010
7383
7398
TCCTGGCGCGGAGCTG
22
2847
|
|
1129538
2017
2032
7405
7420
ACTTTATTGAGTTCCT
9
2848
|
|
1129558
N/A
N/A
126
141
AATCCTGGGACAATCC
77
2849
|
|
1129578
N/A
N/A
207
222
ACATAGGCCTCCTAGT
83
2850
|
|
1129598
N/A
N/A
405
420
GATCTGTTGCTAGTCT
77
2851
|
|
1129618
N/A
N/A
466
481
GGTGGAATCTACAAGG
107
2852
|
|
1129638
N/A
N/A
613
628
CAGGTAGGCACTAGAC
76
2853
|
|
1129658
N/A
N/A
660
675
ACACAGCTCACGATCA
63
2854
|
|
1129678
N/A
N/A
959
974
AAATCCCTCGCCCAGA
62
2855
|
|
1129698
N/A
N/A
1132
1147
GTACCCTTCATCTAAA
83
2856
|
|
1129718
N/A
N/A
1562
1577
CACCAGGGTAAGGTCC
64
2857
|
|
1129738
N/A
N/A
1717
1732
GGATTAGGCAAGGATA
76
2858
|
|
1129758
N/A
N/A
2282
2297
CATCGCGAATAATGAG
49
2859
|
|
1129778
N/A
N/A
2469
2484
CTATAGTGCCATCAGC
56
2860
|
|
1129798
N/A
N/A
2631
2646
CAGTCTACTTAGTGCA
63
2861
|
|
1129818
N/A
N/A
2963
2978
GAAGAGCACTTGCCAC
51
2862
|
|
1129838
N/A
N/A
3404
3419
AAGGTCGCTGTGCATT
58
2863
|
|
1129858
N/A
N/A
3882
3897
AGTAATGAGGCGGGAG
58
2864
|
|
1129878
N/A
N/A
4045
4060
TTGTGACCTTGCTACT
50
2865
|
|
1129898
N/A
N/A
4323
4338
TCCGAGTATCCAGCAA
68
2866
|
|
1129918
N/A
N/A
4366
4381
AGAGGGATAGTGGTCT
88
2867
|
|
1129938
N/A
N/A
4635
4650
ACGCAGTGAGCCACCC
43
2868
|
|
1129958
N/A
N/A
5075
5090
CACGCGGCGCACCGGC
66
2869
|
|
1129978
N/A
N/A
5423
5438
GGAATCTAGCTCGCCC
74
2870
|
|
1129998
N/A
N/A
5483
5498
TCGCCGGCAAGGCTGT
80
2871
|
|
1130018
N/A
N/A
5772
5787
CGTGGAAGCTGCGCTG
75
2872
|
|
1130038
N/A
N/A
5877
5892
GCTCCCGAACCCCAAT
69
2873
|
|
1130058
N/A
N/A
6176
6191
GGCGCAACAGAGCTAA
59
2874
|
|
1130078
N/A
N/A
6368
6383
CGCCTAACCCAGTGAT
71
2875
|
|
1130098
N/A
N/A
6394
6409
ACGATACCAAAGTCGC
68
2876
|
|
1130118
N/A
N/A
6457
6472
GGATTCACCTACACAT
73
2877
|
|
1130138
N/A
N/A
6568
6583
GCCTTTGCAGCCCGGC
98
2878
|
|
1130158
N/A
N/A
6630
6645
ATCCAGAGTCGCAGAA
62
2879
|
|
1130178
N/A
N/A
6754
6769
CGCAGCAAGCCCGAAG
69
2880
|
|
1130198
N/A
N/A
6817
6832
CCGCCCGGACGATGGA
78
2881
|
|
1130218
N/A
N/A
7018
7033
GCTCACCTGGCACGCA
98†
2882
|
|
1130238
N/A
N/A
7096
7111
CGGAAACACGCAGCTC
79†
2883
|
|
TABLE 49
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128738
92
107
N/A
N/A
TCGAAAGTGTTGACTC
15
2884
|
|
1128758
132
147
499
514
GCTTTGTACTTATGCT
63
2885
|
|
1128778
163
178
N/A
N/A
GGTGACAGTGAGAACG
19
2886
|
|
1128798
214
229
3565
3580
TTTGTGGTACAGCTGC
28
2887
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
16
2426
|
|
1128819
245
260
3596
3611
GCTGAGGGCCTGGCCG
59
2888
|
|
1128839
301
316
3808
3823
CAAACAGTATCCCCAT
24
2889
|
|
1128859
424
439
4232
4247
GCAGTGGTTTCCAGTG
17
2890
|
|
1128879
494
509
4439
4454
CAGTTCTATACCATAT
15
2891
|
|
1128899
582
597
4666
4681
CACGGGTTGGTGCGGC
33
2892
|
|
1128919
604
619
4688
4703
TAGGCAGCGACCCCCA
31
2893
|
|
1128939
645
660
4729
4744
TAGCCCACCGGGCAGT
40
2894
|
|
1128959
690
705
4919
4934
TCATAGCAGCTTGCCT
22
2895
|
|
1128979
744
759
4973
4988
GGCGCACCCGAGAGCG
32
2896
|
|
1128999
768
783
4997
5012
GCCTCCGAGGCCCACG
46
2897
|
|
1129019
813
828
5042
5057
AGTCCCCAGTTCCGCG
39
2898
|
|
1129039
871
886
5185
5200
CACGAAGCACCACGGG
16
2899
|
|
1129059
908
923
5222
5237
GGTCGCAGTACTCCCA
18
2900
|
|
1129079
996
1011
5310
5325
GCCGGCTGCGCGGGCA
35
2901
|
|
1129099
1064
1079
N/A
N/A
TCGCCGGCAAGGCTCC
36
2902
|
|
1129119
1107
1122
5526
5541
CTCAGTGGGCCGTTCC
17
2903
|
|
1129139
1135
1150
5554
5569
CAGACTCTTGCGGAGC
12
2904
|
|
1129159
1159
1174
5578
5593
GCCAACGACGCGGGTC
27
2905
|
|
1129179
1222
1237
5641
5656
GAAACTGTGGCCCCAG
18
2906
|
|
1129199
1286
1301
5705
5720
GCCGGTCCTGCAGGCA
33
2907
|
|
1129219
1353
1368
6013
6028
TGGCACGGCTCACAGC
27
2908
|
|
1129239
1403
1418
6063
6078
TGACGGGCGAGAAGGC
47
2909
|
|
1129259
1445
1460
6188
6203
CATCCTCCTGAAGGCG
32
2910
|
|
1129279
1470
1485
6213
6228
GACAGGAGCGCGCAGC
27
2911
|
|
1129299
1505
1520
6248
6263
CGCCGCTTGGCAGGCA
59
2912
|
|
1129319
1564
1579
6307
6322
CTCGAACTGGTGGCCC
37
2913
|
|
1129339
1592
1607
6890
6905
GCAGGAAGCTGGCATA
72
2914
|
|
1129359
1660
1675
6958
6973
GATGGAGGATCCGTGC
11†
2915
|
|
1129379
1699
1714
6997
7012
GCCGCCCTCGAGGAAC
6†
2916
|
|
1129399
1754
1769
7142
7157
CAGCTTGGTCCTCACA
26†
2917
|
|
1129419
1808
1823
7196
7211
CACCACAGCCCGATCC
37
2918
|
|
1129439
1845
1860
7233
7248
TAGGCCACATCGGTGT
31
2919
|
|
1129459
1879
1894
7267
7282
GGAAACGGTGTGCTCC
41
2920
|
|
1129479
1904
1919
7292
7307
GGAAAGATGAGTCCCT
15
2921
|
|
1129499
1951
1966
7339
7354
TGCCTTCCATGCCCCA
8
2922
|
|
1129519
1996
2011
7384
7399
ATCCTGGCGCGGAGCT
29
2923
|
|
1129539
2018
2033
7406
7421
CACTTTATTGAGTTCC
6
2924
|
|
1129559
N/A
N/A
138
153
ACCCCCCAGAACAATC
45
2925
|
|
1129579
N/A
N/A
208
223
CACATAGGCCTCCTAG
24
2926
|
|
1129599
N/A
N/A
408
423
ACCGATCTGTTGCTAG
79
2927
|
|
1129619
N/A
N/A
469
484
CAAGGTGGAATCTACA
50
2928
|
|
1129639
N/A
N/A
614
629
CCAGGTAGGCACTAGA
36
2929
|
|
1129659
N/A
N/A
693
708
TTAGACACAGCCCATA
56
2930
|
|
1129679
N/A
N/A
960
975
CAAATCCCTCGCCCAG
30
2931
|
|
1129699
N/A
N/A
1134
1149
AGGTACCCTTCATCTA
79
2932
|
|
1129719
N/A
N/A
1583
1598
TCAACCCCACCAAGTC
59
2933
|
|
1129739
N/A
N/A
1757
1772
ATGGGATGGACGGACA
44
2934
|
|
1129759
N/A
N/A
2283
2298
CCATCGCGAATAATGA
50
2935
|
|
1129779
N/A
N/A
2470
2485
GCTATAGTGCCATCAG
25
2936
|
|
1129799
N/A
N/A
2651
2666
TATTGAATGAGTGTCC
36
2937
|
|
1129819
N/A
N/A
2987
3002
GTAAGTCTTCAGTCCC
56
2938
|
|
1129839
N/A
N/A
3405
3420
AAAGGTCGCTGTGCAT
58
2939
|
|
1129859
N/A
N/A
3883
3898
GAGTAATGAGGCGGGA
55
2940
|
|
1129879
N/A
N/A
4058
4073
GAACCTACTTGCCTTG
42
2941
|
|
1129899
N/A
N/A
4324
4339
CTCCGAGTATCCAGCA
28
2942
|
|
1129919
N/A
N/A
4367
4382
AAGAGGGATAGTGGTC
48
2943
|
|
1129939
N/A
N/A
4636
4651
AACGCAGTGAGCCACC
55
2944
|
|
1129959
N/A
N/A
5076
5091
CCACGCGGCGCACCGG
61
2945
|
|
1129979
N/A
N/A
5424
5439
CGGAATCTAGCTCGCC
45
2946
|
|
1129999
N/A
N/A
5484
5499
TTCGCCGGCAAGGCTG
50
2947
|
|
1130019
N/A
N/A
5800
5815
AGAAGGTAGGGCACGG
78
2948
|
|
1130039
N/A
N/A
6089
6104
CCACGCACCCAGGTCG
87
2949
|
|
1130059
N/A
N/A
6177
6192
AGGCGCAACAGAGCTA
32
2950
|
|
1130079
N/A
N/A
6369
6384
CCGCCTAACCCAGTGA
28
2951
|
|
1130099
N/A
N/A
6395
6410
AACGATACCAAAGTCG
42
2952
|
|
1130119
N/A
N/A
6459
6474
TGGGATTCACCTACAC
94
2953
|
|
1130139
N/A
N/A
6576
6591
CCTAGTTGGCCTTTGC
46
2954
|
|
1130159
N/A
N/A
6634
6649
ACCCATCCAGAGTCGC
66
2955
|
|
1130179
N/A
N/A
6755
6770
TCGCAGCAAGCCCGAA
53
2956
|
|
1130199
N/A
N/A
6818
6833
GCCGCCCGGACGATGG
84
2957
|
|
1130219
N/A
N/A
7026
7041
GGCTAAGAGCTCACCT
81†
2958
|
|
1130239
N/A
N/A
7097
7112
TCGGAAACACGCAGCT
65†
2959
|
|
TABLE 50
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128739
93
108
N/A
N/A
ATCGAAAGTGTTGACT
23
2960
|
|
1128759
133
148
500
515
AGCTTTGTACTTATGC
13
2961
|
|
1128779
164
179
3515
3530
CGGTGACAGTGAGAAC
14
2962
|
|
1128799
215
230
3566
3581
ATTTGTGGTACAGCTG
7
2963
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
4
2426
|
|
1128820
257
272
N/A
N/A
TAGCACACCAGGGCTG
40
2964
|
|
1128840
302
317
3809
3824
CCAAACAGTATCCCCA
10
2965
|
|
1128860
431
446
4239
4254
CTTTCTGGCAGTGGTT
12
2966
|
|
1128880
497
512
4442
4457
GCTCAGTTCTATACCA
2
2967
|
|
1128900
583
598
4667
4682
GCACGGGTTGGTGCGG
38
2968
|
|
1128920
607
622
4691
4706
CTCTAGGCAGCGACCC
18
2969
|
|
1128940
646
661
4730
4745
GTAGCCCACCGGGCAG
33
2970
|
|
1128960
691
706
4920
4935
ATCATAGCAGCTTGCC
15
2971
|
|
1128980
745
760
4974
4989
GGGCGCACCCGAGAGC
10
2972
|
|
1129000
776
791
5005
5020
GGTAGGTGGCCTCCGA
10
2973
|
|
1129020
814
829
5043
5058
CAGTCCCCAGTTCCGC
8
2974
|
|
1129040
872
887
5186
5201
GCACGAAGCACCACGG
18
2975
|
|
1129060
910
925
5224
5239
CAGGTCGCAGTACTCC
24
2976
|
|
1129080
998
1013
5312
5327
GTGCCGGCTGCGCGGG
15
2977
|
|
1129100
1065
1080
N/A
N/A
TTCGCCGGCAAGGCTC
36
2978
|
|
1129120
1108
1123
5527
5542
GCTCAGTGGGCCGTTC
10
2979
|
|
1129140
1136
1151
5555
5570
ACAGACTCTTGCGGAG
16
2980
|
|
1129160
1160
1175
5579
5594
CGCCAACGACGCGGGT
21
2981
|
|
1129180
1223
1238
5642
5657
AGAAACTGTGGCCCCA
10
2982
|
|
1129200
1288
1303
N/A
N/A
GGGCCGGTCCTGCAGG
15
2983
|
|
1129220
1354
1369
6014
6029
CTGGCACGGCTCACAG
23
2984
|
|
1129240
1404
1419
6064
6079
CTGACGGGCGAGAAGG
46
2985
|
|
1129260
1446
1461
6189
6204
GCATCCTCCTGAAGGC
10
2986
|
|
1129280
1471
1486
6214
6229
CGACAGGAGCGCGCAG
39
2987
|
|
1129300
1518
1533
6261
6276
GAGGGTCGCGCGGCGC
24
2988
|
|
1129320
1565
1580
6308
6323
CCTCGAACTGGTGGCC
48
2989
|
|
1129340
1602
1617
6900
6915
TGCGCCTCCTGCAGGA
70
2990
|
|
1129360
1662
1677
6960
6975
AGGATGGAGGATCCGT
43†
2991
|
|
1129380
1700
1715
6998
7013
TGCCGCCCTCGAGGAA
10†
2992
|
|
1129400
1755
1770
7143
7158
GCAGCTTGGTCCTCAC
8†
2993
|
|
1129420
1811
1826
7199
7214
GGTCACCACAGCCCGA
9
2994
|
|
1129440
1846
1861
7234
7249
GTAGGCCACATCGGTG
12
2995
|
|
1129460
1880
1895
7268
7283
AGGAAACGGTGTGCTC
7
2996
|
|
1129480
1905
1920
7293
7308
GGGAAAGATGAGTCCC
39
2997
|
|
1129500
1952
1967
7340
7355
TTGCCTTCCATGCCCC
4
2998
|
|
1129520
1997
2012
7385
7400
CATCCTGGCGCGGAGC
15
2999
|
|
1129540
2019
2034
7407
7422
GCACTTTATTGAGTTC
3
3000
|
|
1129560
N/A
N/A
139
154
GACCCCCCAGAACAAT
20
3001
|
|
1129580
N/A
N/A
216
231
CACCTTTCCACATAGG
44
3002
|
|
1129600
N/A
N/A
409
424
CACCGATCTGTTGCTA
77
3003
|
|
1129620
N/A
N/A
470
485
CCAAGGTGGAATCTAC
34
3004
|
|
1129640
N/A
N/A
615
630
ACCAGGTAGGCACTAG
23
3005
|
|
1129660
N/A
N/A
694
709
CTTAGACACAGCCCAT
58
3006
|
|
1129680
N/A
N/A
963
978
ACTCAAATCCCTCGCC
23
3007
|
|
1129700
N/A
N/A
1136
1151
GCAGGTACCCTTCATC
22
3008
|
|
1129720
N/A
N/A
1586
1601
GACTCAACCCCACCAA
32
3009
|
|
1129740
N/A
N/A
1758
1773
GATGGGATGGACGGAC
16
3010
|
|
1129760
N/A
N/A
2284
2299
ACCATCGCGAATAATG
14
3011
|
|
1129780
N/A
N/A
2487
2502
ACTTGATTCAGGTGTG
29
3012
|
|
1129800
N/A
N/A
2652
2667
CTATTGAATGAGTGTC
47
3013
|
|
1129820
N/A
N/A
2989
3004
GGGTAAGTCTTCAGTC
33
3014
|
|
1129840
N/A
N/A
3406
3421
AAAAGGTCGCTGTGCA
49
3015
|
|
1129860
N/A
N/A
3884
3899
AGAGTAATGAGGCGGG
27
3016
|
|
1129880
N/A
N/A
4059
4074
TGAACCTACTTGCCTT
21
3017
|
|
1129900
N/A
N/A
4325
4340
TCTCCGAGTATCCAGC
34
3018
|
|
1129920
N/A
N/A
4529
4544
CGTTCCCAACCATCTG
56
3019
|
|
1129940
N/A
N/A
4637
4652
GAACGCAGTGAGCCAC
21
3020
|
|
1129960
N/A
N/A
5078
5093
CCCCACGCGGCGCACC
44
3021
|
|
1129980
N/A
N/A
5425
5440
CCGGAATCTAGCTCGC
32
3022
|
|
1130000
N/A
N/A
5706
5721
CGCCGGTCCTGCAGGC
32
3023
|
|
1130020
N/A
N/A
5801
5816
GAGAAGGTAGGGCACG
30
3024
|
|
1130040
N/A
N/A
6112
6127
CTTCCCGTCCCCGCGG
45
3025
|
|
1130060
N/A
N/A
6178
6193
AAGGCGCAACAGAGCT
19
3026
|
|
1130080
N/A
N/A
6370
6385
TCCGCCTAACCCAGTG
76
3027
|
|
1130100
N/A
N/A
6396
6411
GAACGATACCAAAGTC
38
3028
|
|
1130120
N/A
N/A
6462
6477
ACCTGGGATTCACCTA
31
3029
|
|
1130140
N/A
N/A
6577
6592
TCCTAGTTGGCCTTTG
23
3030
|
|
1130160
N/A
N/A
6659
6674
CGGAAACAGAAACCCC
57
3031
|
|
1130180
N/A
N/A
6756
6771
CTCGCAGCAAGCCCGA
46
3032
|
|
1130200
N/A
N/A
6819
6834
TGCCGCCCGGACGATG
26
3033
|
|
1130220
N/A
N/A
7028
7043
CGGGCTAAGAGCTCAC
26†
3034
|
|
1130240
N/A
N/A
7098
7113
GTCGGAAACACGCAGC
15†
3035
|
|
TABLE 51
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128740
94
109
N/A
N/A
AATCGAAAGTGTTGAC
55
3036
|
|
1128760
143
158
510
525
TGTGCTCTTCAGCTTT
23
3037
|
|
1128780
165
180
3516
3531
CCGGTGACAGTGAGAA
29
3038
|
|
1128800
216
231
3567
3582
CATTTGTGGTACAGCT
20
3039
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
16
2426
|
|
1128821
261
276
N/A
N/A
GTGGTAGCACACCAGG
19
3040
|
|
1128841
303
318
3810
3825
TCCAAACAGTATCCCC
16
3041
|
|
1128861
432
447
N/A
N/A
TCTTTCTGGCAGTGGT
31
3042
|
|
1128881
528
543
4473
4488
CCCTTGCACTGGCATC
17
3043
|
|
1128901
584
599
4668
4683
GGCACGGGTTGGTGCG
69
3044
|
|
1128921
608
623
4692
4707
CCTCTAGGCAGCGACC
32
3045
|
|
1128941
648
663
4732
4747
GTGTAGCCCACCGGGC
84
3046
|
|
1128961
692
707
4921
4936
CATCATAGCAGCTTGC
28
3047
|
|
1128981
746
761
4975
4990
AGGGCGCACCCGAGAG
25
3048
|
|
1129001
777
792
5006
5021
CGGTAGGTGGCCTCCG
57
3049
|
|
1129021
815
830
5044
5059
CCAGTCCCCAGTTCCG
39
3050
|
|
1129041
873
888
5187
5202
AGCACGAAGCACCACG
25
3051
|
|
1129061
911
926
5225
5240
CCAGGTCGCAGTACTC
21
3052
|
|
1129081
1001
1016
5315
5330
GCGGTGCCGGCTGCGC
73
3053
|
|
1129101
1066
1081
5485
5500
CTTCGCCGGCAAGGCT
36
3054
|
|
1129121
1112
1127
5531
5546
CGCAGCTCAGTGGGCC
46
3055
|
|
1129141
1137
1152
5556
5571
GACAGACTCTTGCGGA
23
3056
|
|
1129161
1161
1176
5580
5595
CCGCCAACGACGCGGG
38
3057
|
|
1129181
1224
1239
5643
5658
CAGAAACTGTGGCCCC
29
3058
|
|
1129201
1289
1304
N/A
N/A
CGGGCCGGTCCTGCAG
58
3059
|
|
1129221
1357
1372
6017
6032
CGTCTGGCACGGCTCA
12
3060
|
|
1129241
1405
1420
6065
6080
GCTGACGGGCGAGAAG
30
3061
|
|
1129261
1448
1463
6191
6206
CCGCATCCTCCTGAAG
46
3062
|
|
1129281
1472
1487
6215
6230
GCGACAGGAGCGCGCA
59
3063
|
|
1129301
1519
1534
6262
6277
GGAGGGTCGCGCGGCG
61
3064
|
|
1129321
1566
1581
6309
6324
CCCTCGAACTGGTGGC
70
3065
|
|
1129341
1606
1621
6904
6919
TACCTGCGCCTCCTGC
90
3066
|
|
1129361
1663
1678
6961
6976
GAGGATGGAGGATCCG
43†
3067
|
|
1129381
1704
1719
7002
7017
TCGGTGCCGCCCTCGA
32†
3068
|
|
1129401
1759
1774
7147
7162
CTCTGCAGCTTGGTCC
32†
3069
|
|
1129421
1814
1829
7202
7217
TGCGGTCACCACAGCC
22
3070
|
|
1129441
1847
1862
7235
7250
AGTAGGCCACATCGGT
13
3071
|
|
1129461
1881
1896
7269
7284
CAGGAAACGGTGTGCT
25
3072
|
|
1129481
1906
1921
7294
7309
AGGGAAAGATGAGTCC
10
3073
|
|
1129501
1954
1969
7342
7357
TCTTGCCTTCCATGCC
14
3074
|
|
1129521
1998
2013
7386
7401
CCATCCTGGCGCGGAG
28
3075
|
|
1129541
2020
2035
7408
7423
AGCACTTTATTGAGTT
8
3076
|
|
1129561
N/A
N/A
140
155
CGACCCCCCAGAACAA
72
3077
|
|
1129581
N/A
N/A
229
244
TCCGGGCTGGCCTCAC
81
3078
|
|
1129601
N/A
N/A
410
425
CCACCGATCTGTTGCT
56
3079
|
|
1129621
N/A
N/A
520
535
TTACCGACTGTGTGCT
30
3080
|
|
1129641
N/A
N/A
619
634
TAGCACCAGGTAGGCA
64
3081
|
|
1129661
N/A
N/A
697
712
GCACTTAGACACAGCC
88
3082
|
|
1129681
N/A
N/A
964
979
AACTCAAATCCCTCGC
70
3083
|
|
1129701
N/A
N/A
1137
1152
GGCAGGTACCCTTCAT
151
3084
|
|
1129721
N/A
N/A
1590
1605
TAGAGACTCAACCCCA
62
3085
|
|
1129741
N/A
N/A
1761
1776
GCAGATGGGATGGACG
42
3086
|
|
1129761
N/A
N/A
2285
2300
AACCATCGCGAATAAT
50
3087
|
|
1129781
N/A
N/A
2489
2504
ACACTTGATTCAGGTG
49
3088
|
|
1129801
N/A
N/A
2653
2668
CCTATTGAATGAGTGT
54
3089
|
|
1129821
N/A
N/A
2991
3006
TTGGGTAAGTCTTCAG
51
3090
|
|
1129841
N/A
N/A
3407
3422
AAAAAGGTCGCTGTGC
52
3091
|
|
1129861
N/A
N/A
3885
3900
GAGAGTAATGAGGCGG
54
3092
|
|
1129881
N/A
N/A
4060
4075
TTGAACCTACTTGCCT
62
3093
|
|
1129901
N/A
N/A
4326
4341
GTCTCCGAGTATCCAG
106
3094
|
|
1129921
N/A
N/A
4532
4547
GCCCGTTCCCAACCAT
63
3095
|
|
1129941
N/A
N/A
4639
4654
GGGAACGCAGTGAGCC
47
3096
|
|
1129961
N/A
N/A
5080
5095
AGCCCCACGCGGCGCA
112
3097
|
|
1129981
N/A
N/A
5426
5441
GCCGGAATCTAGCTCG
49
3098
|
|
1130001
N/A
N/A
5707
5722
TCGCCGGTCCTGCAGG
60
3099
|
|
1130021
N/A
N/A
5806
5821
GGCGGGAGAAGGTAGG
51
3100
|
|
1130041
N/A
N/A
6158
6173
CGGGCGGAGAGGAGCG
118
3101
|
|
1130061
N/A
N/A
6310
6325
ACCCTCGAACTGGTGG
45
3102
|
|
1130081
N/A
N/A
6371
6386
TTCCGCCTAACCCAGT
113
3103
|
|
1130101
N/A
N/A
6397
6412
GGAACGATACCAAAGT
75
3104
|
|
1130121
N/A
N/A
6463
6478
TACCTGGGATTCACCT
59
3105
|
|
1130141
N/A
N/A
6578
6593
GTCCTAGTTGGCCTTT
29
3106
|
|
1130161
N/A
N/A
6664
6679
CGGAGCGGAAACAGAA
109
3107
|
|
1130181
N/A
N/A
6757
6772
TCTCGCAGCAAGCCCG
99
3108
|
|
1130201
N/A
N/A
6823
6838
CGCTTGCCGCCCGGAC
48
3109
|
|
1130221
N/A
N/A
7029
7044
CCGGGCTAAGAGCTCA
48†
3110
|
|
1130241
N/A
N/A
7099
7114
GGTCGGAAACACGCAG
77†
3111
|
|
TABLE 52
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128761
145
160
512
527
TGTGTGCTCTTCAGCT
19
3112
|
|
1128781
166
181
3517
3532
CCCGGTGACAGTGAGA
18
3113
|
|
1128801
217
232
3568
3583
ACATTTGTGGTACAGC
8
3114
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
9
2426
|
|
1128862
462
477
4407
4422
AACCGGAGAAGCTGAG
24
3115
|
|
1128882
529
544
4474
4489
ACCCTTGCACTGGCAT
32
3116
|
|
1128902
585
600
4669
4684
AGGCACGGGTTGGTGC
70
3117
|
|
1128922
609
624
4693
4708
ACCTCTAGGCAGCGAC
19
3118
|
|
1128942
650
665
4734
4749
CGGTGTAGCCCACCGG
67
3119
|
|
1128962
693
708
4922
4937
CCATCATAGCAGCTTG
22
3120
|
|
1128982
747
762
4976
4991
CAGGGCGCACCCGAGA
27
3121
|
|
1129002
778
793
5007
5022
CCGGTAGGTGGCCTCC
32
3122
|
|
1129022
823
838
5052
5067
GTGGCCGCCCAGTCCC
17
3123
|
|
1129042
874
889
5188
5203
CAGCACGAAGCACCAC
21
3124
|
|
1129062
912
927
5226
5241
GCCAGGTCGCAGTACT
25
3125
|
|
1129082
1003
1018
5317
5332
CGGCGGTGCCGGCTGC
25
3126
|
|
1129102
1068
1083
5487
5502
CGCTTCGCCGGCAAGG
30
3127
|
|
1129122
1113
1128
5532
5547
CCGCAGCTCAGTGGGC
40
3128
|
|
1129142
1138
1153
5557
5572
AGACAGACTCTTGCGG
15
3129
|
|
1129162
1162
1177
5581
5596
CCCGCCAACGACGCGG
43
3130
|
|
1129182
1225
1240
5644
5659
GCAGAAACTGTGGCCC
33
3131
|
|
1129222
1371
1386
6031
6046
GAGCGCACGGCCAACG
23
3132
|
|
1129242
1406
1421
6066
6081
AGCTGACGGGCGAGAA
59
3133
|
|
1129262
1449
1464
6192
6207
TCCGCATCCTCCTGAA
55
3134
|
|
1129282
1473
1488
6216
6231
GGCGACAGGAGCGCGC
34
3135
|
|
1129302
1520
1535
6263
6278
CGGAGGGTCGCGCGGC
65
3136
|
|
1129342
1607
1622
6905
6920
GTACCTGCGCCTCCTG
82
3137
|
|
1129362
1679
1694
6977
6992
CGCAGAGCATGCCGGG
6†
3138
|
|
1129382
1705
1720
7003
7018
ATCGGTGCCGCCCTCG
8†
3139
|
|
1129402
1774
1789
7162
7177
CAGGGTGAGCCGGCGC
31
3140
|
|
1129422
1815
1830
7203
7218
TTGCGGTCACCACAGC
27
3141
|
|
1129442
1848
1863
7236
7251
TAGTAGGCCACATCGG
19
3142
|
|
1129462
1882
1897
7270
7285
TCAGGAAACGGTGTGC
18
3143
|
|
1129482
1908
1923
7296
7311
GGAGGGAAAGATGAGT
15
3144
|
|
1129502
1955
1970
7343
7358
ATCTTGCCTTCCATGC
9
3145
|
|
1129522
1999
2014
7387
7402
GCCATCCTGGCGCGGA
51
3146
|
|
1129542
2021
2036
7409
7424
AAGCACTTTATTGAGT
12
3147
|
|
1129562
N/A
N/A
141
156
GCGACCCCCCAGAACA
72
3148
|
|
1129582
N/A
N/A
230
245
TTCCGGGCTGGCCTCA
52
3149
|
|
1129602
N/A
N/A
411
426
GCCACCGATCTGTTGC
82
3150
|
|
1129622
N/A
N/A
521
536
CTTACCGACTGTGTGC
52
3151
|
|
1129642
N/A
N/A
623
638
GACCTAGCACCAGGTA
103
3152
|
|
1129662
N/A
N/A
699
714
GGGCACTTAGACACAG
55
3153
|
|
1129682
N/A
N/A
965
980
GAACTCAAATCCCTCG
49
3154
|
|
1129702
N/A
N/A
1138
1153
TGGCAGGTACCCTTCA
62
3155
|
|
1129722
N/A
N/A
1591
1606
TTAGAGACTCAACCCC
62
3156
|
|
1129742
N/A
N/A
1916
1931
CATGAAGTTGTGTGCC
67
3157
|
|
1129762
N/A
N/A
2287
2302
ATAACCATCGCGAATA
36
3158
|
|
1129782
N/A
N/A
2490
2505
GACACTTGATTCAGGT
45
3159
|
|
1129802
N/A
N/A
2654
2669
TCCTATTGAATGAGTG
23
3160
|
|
1129822
N/A
N/A
3016
3031
TCCACAACCTGCTAGC
74
3161
|
|
1129842
N/A
N/A
3408
3423
CAAAAAGGTCGCTGTG
59
3162
|
|
1129862
N/A
N/A
3886
3901
GGAGAGTAATGAGGCG
44
3163
|
|
1129882
N/A
N/A
4061
4076
CTTGAACCTACTTGCC
50
3164
|
|
1129902
N/A
N/A
4328
4343
AAGTCTCCGAGTATCC
61
3165
|
|
1129922
N/A
N/A
4534
4549
TGGCCCGTTCCCAACC
46
3166
|
|
1129942
N/A
N/A
4805
4820
AAGCCCGTCCCACCTG
46
3167
|
|
1129962
N/A
N/A
5084
5099
ACCCAGCCCCACGCGG
60
3168
|
|
1129982
N/A
N/A
5428
5443
TGGCCGGAATCTAGCT
119
3169
|
|
1130002
N/A
N/A
5708
5723
CTCGCCGGTCCTGCAG
54
3170
|
|
1130022
N/A
N/A
5823
5838
CGTGGAAAGAAGGGTG
78
3171
|
|
1130042
N/A
N/A
6159
6174
CCGGGCGGAGAGGAGC
87
3172
|
|
1130062
N/A
N/A
6311
6326
TACCCTCGAACTGGTG
69
3173
|
|
1130082
N/A
N/A
6373
6388
TCTTCCGCCTAACCCA
74
3174
|
|
1130102
N/A
N/A
6398
6413
CGGAACGATACCAAAG
47
3175
|
|
1130122
N/A
N/A
6464
6479
CTACCTGGGATTCACC
85
3176
|
|
1130142
N/A
N/A
6579
6594
CGTCCTAGTTGGCCTT
33
3177
|
|
1130162
N/A
N/A
6665
6680
GCGGAGCGGAAACAGA
67
3178
|
|
1130182
N/A
N/A
6759
6774
TCTCTCGCAGCAAGCC
47
3179
|
|
1130202
N/A
N/A
6836
6851
CCCCATCTGACAACGC
103
3180
|
|
1130222
N/A
N/A
7030
7045
ACCGGGCTAAGAGCTC
69†
3181
|
|
1130242
N/A
N/A
7100
7115
GGGTCGGAAACACGCA
94†
3182
|
|
1128741
95
110
N/A
N/A
GAATCGAAAGTGTTGA
28
3183
|
|
1128822
263
278
N/A
N/A
GGGTGGTAGCACACCA
36
3184
|
|
1128842
327
342
N/A
N/A
CAGTGGTCTTTCACTT
91
3185
|
|
1129202
1291
1306
N/A
N/A
TGCGGGCCGGTCCTGC
63
3186
|
|
1129322
1567
1582
N/A
N/A
CCCCTCGAACTGGTGG
86
3187
|
|
TABLE 53
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
No.
|
|
1128722
27
42
27
42
GTCCGTTGGTCCAGCT
34
3188
|
|
1128742
96
111
N/A
N/A
GGAATCGAAAGTGTTG
19
3189
|
|
1128762
146
161
513
528
CTGTGTGCTCTTCAGC
11
3190
|
|
1128782
168
183
3519
3534
TCCCCGGTGACAGTGA
16
3191
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
7
2426
|
|
1128803
219
234
3570
3585
GTACATTTGTGGTACA
41
3192
|
|
1128823
278
293
3785
3800
CCTGATCAAAGTTGGG
25
3193
|
|
1128843
328
343
N/A
N/A
GCAGTGGTCTTTCACT
34
3194
|
|
1128863
463
478
4408
4423
AAACCGGAGAAGCTGA
34
3195
|
|
1128883
531
546
4476
4491
GGACCCTTGCACTGGC
33
3196
|
|
1128903
586
601
4670
4685
GAGGCACGGGTTGGTG
33
3197
|
|
1128923
610
625
4694
4709
CACCTCTAGGCAGCGA
26
3198
|
|
1128943
651
666
4735
4750
CCGGTGTAGCCCACCG
45
3199
|
|
1128963
695
710
4924
4939
GGCCATCATAGCAGCT
35
3200
|
|
1128983
748
763
4977
4992
ACAGGGCGCACCCGAG
19
3201
|
|
1129003
779
794
5008
5023
TCCGGTAGGTGGCCTC
32
3202
|
|
1129023
838
853
N/A
N/A
GTTCCGGCAGAAGGCG
20
3203
|
|
1129043
877
892
5191
5206
GTTCAGCACGAAGCAC
14
3204
|
|
1129063
913
928
5227
5242
TGCCAGGTCGCAGTAC
37
3205
|
|
1129083
1004
1019
5318
5333
TCGGCGGTGCCGGCTG
18
3206
|
|
1129103
1069
1084
5488
5503
CCGCTTCGCCGGCAAG
44
3207
|
|
1129123
1114
1129
5533
5548
CCCGCAGCTCAGTGGG
35
3208
|
|
1129143
1139
1154
5558
5573
AAGACAGACTCTTGCG
16
3209
|
|
1129163
1163
1178
5582
5597
GCCCGCCAACGACGCG
39
3210
|
|
1129183
1226
1241
5645
5660
CGCAGAAACTGTGGCC
36
3211
|
|
1129203
1292
1307
N/A
N/A
GTGCGGGCCGGTCCTG
62
3212
|
|
1129223
1373
1388
6033
6048
AGGAGCGCACGGCCAA
27
3213
|
|
1129243
1411
1426
6071
6086
CTGGTAGCTGACGGGC
24
3214
|
|
1129263
1450
1465
6193
6208
GTCCGCATCCTCCTGA
40
3215
|
|
1129283
1474
1489
6217
6232
AGGCGACAGGAGCGCG
20
3216
|
|
1129303
1521
1536
6264
6279
TCGGAGGGTCGCGCGG
48
3217
|
|
1129323
1568
1583
N/A
N/A
CCCCCTCGAACTGGTG
58
3218
|
|
1129343
1609
1624
6907
6922
CGGTACCTGCGCCTCC
87
3219
|
|
1129363
1680
1695
6978
6993
GCGCAGAGCATGCCGG
44†
3220
|
|
1129383
1706
1721
7004
7019
CATCGGTGCCGCCCTC
15†
3221
|
|
1129403
1775
1790
7163
7178
GCAGGGTGAGCCGGCG
27
3222
|
|
1129423
1817
1832
7205
7220
TGTTGCGGTCACCACA
11
3223
|
|
1129443
1849
1864
7237
7252
GTAGTAGGCCACATCG
20
3224
|
|
1129463
1883
1898
7271
7286
ATCAGGAAACGGTGTG
38
3225
|
|
1129483
1919
1934
7307
7322
GGAATCACCAAGGAGG
5
3226
|
|
1129503
1957
1972
7345
7360
CAATCTTGCCTTCCAT
14
3227
|
|
1129523
2000
2015
7388
7403
CGCCATCCTGGCGCGG
57
3228
|
|
1129543
2022
2037
7410
7425
AAAGCACTTTATTGAG
9
3229
|
|
1129563
N/A
N/A
142
157
AGCGACCCCCCAGAAC
42
3230
|
|
1129583
N/A
N/A
232
247
CCTTCCGGGCTGGCCT
55
3231
|
|
1129603
N/A
N/A
412
427
TGCCACCGATCTGTTG
60
3232
|
|
1129623
N/A
N/A
526
541
GGCCACTTACCGACTG
78
3233
|
|
1129643
N/A
N/A
624
639
AGACCTAGCACCAGGT
53
3234
|
|
1129663
N/A
N/A
701
716
GTGGGCACTTAGACAC
63
3235
|
|
1129683
N/A
N/A
1003
1018
TCAACCTGGTTTCACA
20
3236
|
|
1129703
N/A
N/A
1139
1154
ATGGCAGGTACCCTTC
21
3237
|
|
1129723
N/A
N/A
1592
1607
CTTAGAGACTCAACCC
36
3238
|
|
1129743
N/A
N/A
1917
1932
GCATGAAGTTGTGTGC
72
3239
|
|
1129763
N/A
N/A
2288
2303
CATAACCATCGCGAAT
29
3240
|
|
1129783
N/A
N/A
2494
2509
GTTAGACACTTGATTC
21
3241
|
|
1129803
N/A
N/A
2655
2670
CTCCTATTGAATGAGT
64
3242
|
|
1129823
N/A
N/A
3031
3046
GAGTAGACTCCTGACT
80
3243
|
|
1129843
N/A
N/A
3409
3424
TCAAAAAGGTCGCTGT
64
3244
|
|
1129863
N/A
N/A
3895
3910
GTGATACCAGGAGAGT
36
3245
|
|
1129883
N/A
N/A
4062
4077
TCTTGAACCTACTTGC
32
3246
|
|
1129903
N/A
N/A
4330
4345
CCAAGTCTCCGAGTAT
68
3247
|
|
1129923
N/A
N/A
4535
4550
CTGGCCCGTTCCCAAC
41
3248
|
|
1129943
N/A
N/A
4808
4823
GGCAAGCCCGTCCCAC
21
3249
|
|
1129963
N/A
N/A
5148
5163
CTGTAGCCACACGACG
85
3250
|
|
1129983
N/A
N/A
5429
5444
CTGGCCGGAATCTAGC
92
3251
|
|
1130003
N/A
N/A
5709
5724
ACTCGCCGGTCCTGCA
36
3252
|
|
1130023
N/A
N/A
5841
5856
CCGGGAGCTCCGGAGG
69
3253
|
|
1130043
N/A
N/A
6160
6175
CCCGGGCGGAGAGGAG
85
3254
|
|
1130063
N/A
N/A
6312
6327
CTACCCTCGAACTGGT
52
3255
|
|
1130083
N/A
N/A
6375
6390
CTTCTTCCGCCTAACC
51
3256
|
|
1130103
N/A
N/A
6399
6414
CCGGAACGATACCAAA
72
3257
|
|
1130123
N/A
N/A
6475
6490
GCCCTGGGATTCTACC
54
3258
|
|
1130143
N/A
N/A
6600
6615
TAGCCCGGAGCGCGGG
61
3259
|
|
1130163
N/A
N/A
6666
6681
GGCGGAGCGGAAACAG
49
3260
|
|
1130183
N/A
N/A
6760
6775
CTCTCTCGCAGCAAGC
43
3261
|
|
1130203
N/A
N/A
6838
6853
CACCCCATCTGACAAC
45
3262
|
|
1130223
N/A
N/A
7031
7046
AACCGGGCTAAGAGCT
62†
3263
|
|
1130243
N/A
N/A
7101
7116
TGGGTCGGAAACACGC
33†
3264
|
|
TABLE 54
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
No.
|
|
1128723
29
44
29
44
CCGTCCGTTGGTCCAG
25
3265
|
|
1128743
97
112
N/A
N/A
TGGAATCGAAAGTGTT
42
3266
|
|
1128763
147
162
514
529
ACTGTGTGCTCTTCAG
20
3267
|
|
1128783
169
184
3520
3535
CTCCCCGGTGACAGTG
36
3268
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
12
2426
|
|
1128804
220
235
3571
3586
GGTACATTTGTGGTAC
39
3269
|
|
1128824
279
294
3786
3801
TCCTGATCAAAGTTGG
32
3270
|
|
1128844
330
345
N/A
N/A
CTGCAGTGGTCTTTCA
87
3271
|
|
1128864
464
479
4409
4424
AAAACCGGAGAAGCTG
32
3272
|
|
1128884
535
550
4480
4495
ATCAGGACCCTTGCAC
32
3273
|
|
1128904
587
602
4671
4686
GGAGGCACGGGTTGGT
41
3274
|
|
1128924
612
627
4696
4711
TCCACCTCTAGGCAGC
26
3275
|
|
1128944
652
667
4736
4751
TCCGGTGTAGCCCACC
31
3276
|
|
1128964
697
712
4926
4941
GCGGCCATCATAGCAG
70
3277
|
|
1128984
749
764
4978
4993
GACAGGGCGCACCCGA
28
3278
|
|
1129004
780
795
5009
5024
TTCCGGTAGGTGGCCT
38
3279
|
|
1129024
839
854
N/A
N/A
GGTTCCGGCAGAAGGC
27
3280
|
|
1129044
881
896
5195
5210
CGCGGTTCAGCACGAA
21
3281
|
|
1129064
941
956
5255
5270
GCGCCGCCTGGGTTGG
48
3282
|
|
1129084
1005
1020
5319
5334
TTCGGCGGTGCCGGCT
17
3283
|
|
1129104
1070
1085
5489
5504
CCCGCTTCGCCGGCAA
20
3284
|
|
1129124
1115
1130
5534
5549
GCCCGCAGCTCAGTGG
39
3285
|
|
1129144
1141
1156
5560
5575
CGAAGACAGACTCTTG
20
3286
|
|
1129164
1165
1180
5584
5599
CAGCCCGCCAACGACG
47
3287
|
|
1129184
1227
1242
5646
5661
GCGCAGAAACTGTGGC
51
3288
|
|
1129204
1302
1317
5962
5977
AGATCCTCGGGTGCGG
22
3289
|
|
1129224
1374
1389
6034
6049
TAGGAGCGCACGGCCA
27
3290
|
|
1129244
1412
1427
6072
6087
GCTGGTAGCTGACGGG
44
3291
|
|
1129264
1452
1467
6195
6210
CCGTCCGCATCCTCCT
24
3292
|
|
1129284
1475
1490
6218
6233
AAGGCGACAGGAGCGC
21
3293
|
|
1129304
1522
1537
6265
6280
CTCGGAGGGTCGCGCG
26
3294
|
|
1129324
1569
1584
N/A
N/A
GCCCCCTCGAACTGGT
110
3295
|
|
1129344
1610
1625
6908
6923
ACGGTACCTGCGCCTC
63
3296
|
|
1129364
1681
1696
6979
6994
TGCGCAGAGCATGCCG
44†
3297
|
|
1129384
1707
1722
7005
7020
GCATCGGTGCCGCCCT
36†
3298
|
|
1129404
1776
1791
7164
7179
TGCAGGGTGAGCCGGC
54
3299
|
|
1129424
1819
1834
7207
7222
CTTGTTGCGGTCACCA
17
3300
|
|
1129444
1850
1865
7238
7253
GGTAGTAGGCCACATC
23
3301
|
|
1129464
1884
1899
7272
7287
AATCAGGAAACGGTGT
28
3302
|
|
1129484
1920
1935
7308
7323
CGGAATCACCAAGGAG
9
3303
|
|
1129504
1958
1973
7346
7361
ACAATCTTGCCTTCCA
15
3304
|
|
1129524
2002
2017
7390
7405
TGCGCCATCCTGGCGC
42
3305
|
|
1129544
2023
2038
7411
7426
CAAAGCACTTTATTGA
21
3306
|
|
1129564
N/A
N/A
143
158
TAGCGACCCCCCAGAA
34
3307
|
|
1129584
N/A
N/A
271
286
GTATCCACCCAGTCTG
67
3308
|
|
1129604
N/A
N/A
415
430
GCCTGCCACCGATCTG
81
3309
|
|
1129624
N/A
N/A
528
543
CAGGCCACTTACCGAC
37
3310
|
|
1129644
N/A
N/A
626
641
TAAGACCTAGCACCAG
69
3311
|
|
1129664
N/A
N/A
702
717
CGTGGGCACTTAGACA
47
3312
|
|
1129684
N/A
N/A
1004
1019
ATCAACCTGGTTTCAC
33
3313
|
|
1129704
N/A
N/A
1203
1218
GTAATTTAGTGTCTGG
38
3314
|
|
1129724
N/A
N/A
1593
1608
GCTTAGAGACTCAACC
60
3315
|
|
1129744
N/A
N/A
2082
2097
TGGACCTTTAAGATGC
26
3316
|
|
1129764
N/A
N/A
2289
2304
ACATAACCATCGCGAA
45
3317
|
|
1129784
N/A
N/A
2499
2514
CGTGTGTTAGACACTT
42
3318
|
|
1129804
N/A
N/A
2656
2671
TCTCCTATTGAATGAG
79
3319
|
|
1129824
N/A
N/A
3032
3047
GGAGTAGACTCCTGAC
85
3320
|
|
1129844
N/A
N/A
3410
3425
CTCAAAAAGGTCGCTG
43
3321
|
|
1129864
N/A
N/A
3897
3912
TGGTGATACCAGGAGA
21
3322
|
|
1129884
N/A
N/A
4066
4081
CCCTTCTTGAACCTAC
58
3323
|
|
1129904
N/A
N/A
4332
4347
TGCCAAGTCTCCGAGT
52
3324
|
|
1129924
N/A
N/A
4583
4598
TGGCACACCACCCGGC
51
3325
|
|
1129944
N/A
N/A
4809
4824
TGGCAAGCCCGTCCCA
41
3326
|
|
1129964
N/A
N/A
5149
5164
CCTGTAGCCACACGAC
61
3327
|
|
1129984
N/A
N/A
5433
5448
CCGGCTGGCCGGAATC
63
3328
|
|
1130004
N/A
N/A
5710
5725
TACTCGCCGGTCCTGC
34
3329
|
|
1130024
N/A
N/A
5842
5857
CCCGGGAGCTCCGGAG
138
3330
|
|
1130044
N/A
N/A
6161
6176
ACCCGGGCGGAGAGGA
38
3331
|
|
1130064
N/A
N/A
6313
6328
CCTACCCTCGAACTGG
62
3332
|
|
1130084
N/A
N/A
6380
6395
GCGGGCTTCTTCCGCC
103
3333
|
|
1130104
N/A
N/A
6400
6415
CCCGGAACGATACCAA
75
3334
|
|
1130124
N/A
N/A
6527
6542
CGATTCTCCCTGTATC
34
3335
|
|
1130144
N/A
N/A
6602
6617
ACTAGCCCGGAGCGCG
56
3336
|
|
1130164
N/A
N/A
6667
6682
GGGCGGAGCGGAAACA
87
3337
|
|
1130184
N/A
N/A
6781
6796
GTAAACCCACTCATGC
63
3338
|
|
1130204
N/A
N/A
6841
6856
TCACACCCCATCTGAC
52
3339
|
|
1130224
N/A
N/A
7032
7047
CAACCGGGCTAAGAGC
74†
3340
|
|
1130244
N/A
N/A
7102
7117
CTGGGTCGGAAACACG
57†
3341
|
|
TABLE 55
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128744
98
113
N/A
N/A
GTGGAATCGAAAGTGT
37
3342
|
|
1128764
148
163
515
530
GACTGTGTGCTCTTCA
12
3343
|
|
1128784
170
185
3521
3536
GCTCCCCGGTGACAGT
44
3344
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
13
2426
|
|
1128805
221
236
3572
3587
GGGTACATTTGTGGTA
35
3345
|
|
1128825
284
299
3791
3806
GCTGGTCCTGATCAAA
31
3346
|
|
1128845
365
380
4173
4188
CACAGGTCCCTCCTTT
41
3347
|
|
1128865
465
480
4410
4425
AAAAACCGGAGAAGCT
57
3348
|
|
1128885
536
551
4481
4496
CATCAGGACCCTTGCA
51
3349
|
|
1128905
589
604
4673
4688
ATGGAGGCACGGGTTG
34
3350
|
|
1128925
619
634
4703
4718
GTGGCCCTCCACCTCT
37
3351
|
|
1128945
653
668
4737
4752
CTCCGGTGTAGCCCAC
51
3352
|
|
1128965
698
713
4927
4942
CGCGGCCATCATAGCA
76
3353
|
|
1128985
750
765
4979
4994
TGACAGGGCGCACCCG
43
3354
|
|
1129005
781
796
5010
5025
GTTCCGGTAGGTGGCC
49
3355
|
|
1129025
840
855
N/A
N/A
GGGTTCCGGCAGAAGG
37
3356
|
|
1129045
882
897
5196
5211
TCGCGGTTCAGCACGA
41
3357
|
|
1129065
973
988
5287
5302
GACATGAAGCCTAGGG
17
3358
|
|
1129085
1006
1021
5320
5335
CTTCGGCGGTGCCGGC
41
3359
|
|
1129105
1071
1086
5490
5505
TCCCGCTTCGCCGGCA
35
3360
|
|
1129125
1117
1132
5536
5551
CTGCCCGCAGCTCAGT
36
3361
|
|
1129145
1142
1157
5561
5576
TCGAAGACAGACTCTT
31
3362
|
|
1129165
1168
1183
5587
5602
CACCAGCCCGCCAACG
53
3363
|
|
1129185
1228
1243
5647
5662
GGCGCAGAAACTGTGG
42
3364
|
|
1129205
1304
1319
5964
5979
TCAGATCCTCGGGTGC
29
3365
|
|
1129225
1375
1390
6035
6050
GTAGGAGCGCACGGCC
32
3366
|
|
1129245
1413
1428
6073
6088
TGCTGGTAGCTGACGG
36
3367
|
|
1129265
1454
1469
6197
6212
TGCCGTCCGCATCCTC
40
3368
|
|
1129285
1476
1491
6219
6234
TAAGGCGACAGGAGCG
33
3369
|
|
1129305
1523
1538
6266
6281
TCTCGGAGGGTCGCGC
39
3370
|
|
1129325
1570
1585
N/A
N/A
CGCCCCCTCGAACTGG
131
3371
|
|
1129345
1611
1626
6909
6924
AACGGTACCTGCGCCT
82
3372
|
|
1129365
1684
1699
6982
6997
CCCTGCGCAGAGCATG
21†
3373
|
|
1129385
1708
1723
7006
7021
CGCATCGGTGCCGCCC
17†
3374
|
|
1129405
1777
1792
7165
7180
TTGCAGGGTGAGCCGG
40
3375
|
|
1129425
1820
1835
7208
7223
GCTTGTTGCGGTCACC
27
3376
|
|
1129445
1851
1866
7239
7254
AGGTAGTAGGCCACAT
23
3377
|
|
1129465
1885
1900
7273
7288
CAATCAGGAAACGGTG
29
3378
|
|
1129485
1921
1936
7309
7324
GCGGAATCACCAAGGA
7
3379
|
|
1129505
1962
1977
7350
7365
GGACACAATCTTGCCT
34
3380
|
|
1129525
2003
2018
7391
7406
CTGCGCCATCCTGGCG
65
3381
|
|
1129545
2024
2039
7412
7427
TCAAAGCACTTTATTG
15
3382
|
|
1129565
N/A
N/A
144
159
ATAGCGACCCCCCAGA
46
3383
|
|
1129585
N/A
N/A
273
288
TTGTATCCACCCAGTC
93
3384
|
|
1129605
N/A
N/A
420
435
TCATAGCCTGCCACCG
73
3385
|
|
1129625
N/A
N/A
554
569
ACCCAAGGGTTCCCGG
142
3386
|
|
1129645
N/A
N/A
629
644
GCATAAGACCTAGCAC
80
3387
|
|
1129665
N/A
N/A
725
740
AAGCACCTGCTCCGAG
78
3388
|
|
1129685
N/A
N/A
1009
1024
CTGTTATCAACCTGGT
64
3389
|
|
1129705
N/A
N/A
1204
1219
GGTAATTTAGTGTCTG
48
3390
|
|
1129725
N/A
N/A
1594
1609
TGCTTAGAGACTCAAC
76
3391
|
|
1129745
N/A
N/A
2084
2099
GATGGACCTTTAAGAT
72
3392
|
|
1129765
N/A
N/A
2290
2305
AACATAACCATCGCGA
55
3393
|
|
1129785
N/A
N/A
2513
2528
CCTTAGGGAGAAAGCG
51
3394
|
|
1129805
N/A
N/A
2659
2674
AGCTCTCCTATTGAAT
69
3395
|
|
1129825
N/A
N/A
3034
3049
CTGGAGTAGACTCCTG
66
3396
|
|
1129845
N/A
N/A
3411
3426
CCTCAAAAAGGTCGCT
59
3397
|
|
1129865
N/A
N/A
3900
3915
GTCTGGTGATACCAGG
54
3398
|
|
1129885
N/A
N/A
4069
4084
AGGCCCTTCTTGAACC
72
3399
|
|
1129905
N/A
N/A
4333
4348
ATGCCAAGTCTCCGAG
68
3400
|
|
1129925
N/A
N/A
4584
4599
CTGGCACACCACCCGG
68
3401
|
|
1129945
N/A
N/A
4810
4825
CTGGCAAGCCCGTCCC
48
3402
|
|
1129965
N/A
N/A
5150
5165
TCCTGTAGCCACACGA
109
3403
|
|
1129985
N/A
N/A
5434
5449
GCCGGCTGGCCGGAAT
89
3404
|
|
1130005
N/A
N/A
5711
5726
GTACTCGCCGGTCCTG
76
3405
|
|
1130025
N/A
N/A
5843
5858
CCCCGGGAGCTCCGGA
77
3406
|
|
1130045
N/A
N/A
6162
6177
AACCCGGGCGGAGAGG
96
3407
|
|
1130065
N/A
N/A
6314
6329
GCCTACCCTCGAACTG
85
3408
|
|
1130085
N/A
N/A
6381
6396
CGCGGGCTTCTTCCGC
88
3409
|
|
1130105
N/A
N/A
6401
6416
ACCCGGAACGATACCA
69
3410
|
|
1130125
N/A
N/A
6529
6544
TTCGATTCTCCCTGTA
77
3411
|
|
1130145
N/A
N/A
6603
6618
CACTAGCCCGGAGCGC
80
3412
|
|
1130165
N/A
N/A
6668
6683
TGGGCGGAGCGGAAAC
80
3413
|
|
1130185
N/A
N/A
6783
6798
TTGTAAACCCACTCAT
82
3414
|
|
1130205
N/A
N/A
6862
6877
TGCGAACACAGAGCGC
113
3415
|
|
1130225
N/A
N/A
7033
7048
CCAACCGGGCTAAGAG
65†
3416
|
|
1130245
N/A
N/A
7103
7118
CCTGGGTCGGAAACAC
80†
3417
|
|
TABLE 56
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128745
99
114
N/A
N/A
GGTGGAATCGAAAGTG
45
3418
|
|
1128765
149
164
516
531
CGACTGTGTGCTCTTC
13
3419
|
|
1128785
171
186
3522
3537
GGCTCCCCGGTGACAG
37
3420
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
14
2426
|
|
1128806
222
237
3573
3588
TGGGTACATTTGTGGT
33
3421
|
|
1128826
285
300
3792
3807
CGCTGGTCCTGATCAA
23
3422
|
|
1128846
366
381
4174
4189
ACACAGGTCCCTCCTT
48
3423
|
|
1128866
466
481
4411
4426
GAAAAACCGGAGAAGC
46
3424
|
|
1128886
537
552
4482
4497
GCATCAGGACCCTTGC
67
3425
|
|
1128906
590
605
4674
4689
CATGGAGGCACGGGTT
29
3426
|
|
1128926
623
638
4707
4722
GGCGGTGGCCCTCCAC
38
3427
|
|
1128946
654
669
4738
4753
GCTCCGGTGTAGCCCA
69
3428
|
|
1128966
699
714
4928
4943
CCGCGGCCATCATAGC
77
3429
|
|
1128986
751
766
4980
4995
CTGACAGGGCGCACCC
38
3430
|
|
1129006
799
814
5028
5043
CGCTTGCTCGGCAGTC
23
3431
|
|
1129026
841
856
N/A
N/A
CGGGTTCCGGCAGAAG
40
3432
|
|
1129046
883
898
5197
5212
GTCGCGGTTCAGCACG
24
3433
|
|
1129066
974
989
5288
5303
GGACATGAAGCCTAGG
23
3434
|
|
1129086
1007
1022
5321
5336
GCTTCGGCGGTGCCGG
33
3435
|
|
1129106
1073
1088
5492
5507
GCTCCCGCTTCGCCGG
52
3436
|
|
1129126
1119
1134
5538
5553
CGCTGCCCGCAGCTCA
31
3437
|
|
1129146
1143
1158
5562
5577
ATCGAAGACAGACTCT
25
3438
|
|
1129166
1171
1186
5590
5605
CGCCACCAGCCCGCCA
45
3439
|
|
1129186
1229
1244
5648
5663
CGGCGCAGAAACTGTG
36
3440
|
|
1129206
1306
1321
5966
5981
CGTCAGATCCTCGGGT
30
3441
|
|
1129226
1377
1392
6037
6052
CGGTAGGAGCGCACGG
25
3442
|
|
1129246
1429
1444
N/A
N/A
CAACAGAGCCAGGTCG
36
3443
|
|
1129266
1455
1470
6198
6213
CTGCCGTCCGCATCCT
42
3444
|
|
1129286
1477
1492
6220
6235
GTAAGGCGACAGGAGC
23
3445
|
|
1129306
1524
1539
6267
6282
GTCTCGGAGGGTCGCG
33
3446
|
|
1129326
1571
1586
N/A
N/A
CCGCCCCCTCGAACTG
69
3447
|
|
1129346
1612
1627
6910
6925
GAACGGTACCTGCGCC
96
3448
|
|
1129366
1685
1700
6983
6998
ACCCTGCGCAGAGCAT
19†
3449
|
|
1129386
1709
1724
7007
7022
ACGCATCGGTGCCGCC
17†
3450
|
|
1129406
1780
1795
7168
7183
GCCTTGCAGGGTGAGC
25
3451
|
|
1129426
1821
1836
7209
7224
GGCTTGTTGCGGTCAC
16
3452
|
|
1129446
1852
1867
7240
7255
CAGGTAGTAGGCCACA
25
3453
|
|
1129466
1888
1903
7276
7291
GAGCAATCAGGAAACG
20
3454
|
|
1129486
1922
1937
7310
7325
TGCGGAATCACCAAGG
9
3455
|
|
1129506
1964
1979
7352
7367
TGGGACACAATCTTGC
25
3456
|
|
1129526
2004
2019
7392
7407
CCTGCGCCATCCTGGC
21
3457
|
|
1129546
2031
2046
7419
7434
AGCATTTTCAAAGCAC
51
3458
|
|
1129566
N/A
N/A
145
160
GATAGCGACCCCCCAG
53
3459
|
|
1129586
N/A
N/A
277
292
GCCCTTGTATCCACCC
80
3460
|
|
1129606
N/A
N/A
422
437
AGTCATAGCCTGCCAC
110
3461
|
|
1129626
N/A
N/A
555
570
CACCCAAGGGTTCCCG
81
3462
|
|
1129646
N/A
N/A
630
645
GGCATAAGACCTAGCA
73
3463
|
|
1129666
N/A
N/A
727
742
TCAAGCACCTGCTCCG
53
3464
|
|
1129686
N/A
N/A
1010
1025
ACTGTTATCAACCTGG
54
3465
|
|
1129706
N/A
N/A
1206
1221
TAGGTAATTTAGTGTC
55
3466
|
|
1129726
N/A
N/A
1596
1611
CATGCTTAGAGACTCA
51
3467
|
|
1129746
N/A
N/A
2092
2107
GGATCTGAGATGGACC
43
3468
|
|
1129766
N/A
N/A
2291
2306
GAACATAACCATCGCG
60
3469
|
|
1129786
N/A
N/A
2519
2534
AGGCTACCTTAGGGAG
64
3470
|
|
1129806
N/A
N/A
2683
2698
CGTCACGCTGCTGCTG
45
3471
|
|
1129826
N/A
N/A
3035
3050
GCTGGAGTAGACTCCT
54
3472
|
|
1129846
N/A
N/A
3412
3427
CCCTCAAAAAGGTCGC
71
3473
|
|
1129866
N/A
N/A
3956
3971
CAAAGGGTATTGTGGA
48
3474
|
|
1129886
N/A
N/A
4306
4321
CTATTCTGTAGGCCCA
62
3475
|
|
1129906
N/A
N/A
4343
4358
GTCTAGGACCATGCCA
95
3476
|
|
1129926
N/A
N/A
4607
4622
ACCCCCCCAGAGAGCT
91
3477
|
|
1129946
N/A
N/A
4812
4827
TCCTGGCAAGCCCGTC
37
3478
|
|
1129966
N/A
N/A
5157
5172
TCCGGGTTCCTGTAGC
60
3479
|
|
1129986
N/A
N/A
5436
5451
CGGCCGGCTGGCCGGA
84
3480
|
|
1130006
N/A
N/A
5712
5727
GGTACTCGCCGGTCCT
50
3481
|
|
1130026
N/A
N/A
5844
5859
TCCCCGGGAGCTCCGG
102
3482
|
|
1130046
N/A
N/A
6163
6178
TAACCCGGGCGGAGAG
88
3483
|
|
1130066
N/A
N/A
6315
6330
TGCCTACCCTCGAACT
100
3484
|
|
1130086
N/A
N/A
6382
6397
TCGCGGGCTTCTTCCG
56
3485
|
|
1130106
N/A
N/A
6402
6417
CACCCGGAACGATACC
67
3486
|
|
1130126
N/A
N/A
6531
6546
AGTTCGATTCTCCCTG
57
3487
|
|
1130146
N/A
N/A
6604
6619
ACACTAGCCCGGAGCG
66
3488
|
|
1130166
N/A
N/A
6669
6684
ATGGGCGGAGCGGAAA
108
3489
|
|
1130186
N/A
N/A
6784
6799
CTTGTAAACCCACTCA
57
3490
|
|
1130206
N/A
N/A
6863
6878
CTGCGAACACAGAGCG
100
3491
|
|
1130226
N/A
N/A
7034
7049
GCCAACCGGGCTAAGA
134†
3492
|
|
1130246
N/A
N/A
7104
7119
CCCTGGGTCGGAAACA
87†
3493
|
|
TABLE 57
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128746
101
116
N/A
N/A
AAGGTGGAATCGAAAG
68
3494
|
|
1128766
150
165
N/A
N/A
ACGACTGTGTGCTCTT
20
3495
|
|
1128786
175
190
3526
3541
GCAGGGCTCCCCGGTG
45
3496
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
38
2426
|
|
1128807
223
238
3574
3589
GTGGGTACATTTGTGG
57
3497
|
|
1128827
287
302
3794
3809
ATCGCTGGTCCTGATC
57
3498
|
|
1128847
370
385
4178
4193
GTTCACACAGGTCCCT
40
3499
|
|
1128867
467
482
4412
4427
GGAAAAACCGGAGAAG
63
3500
|
|
1128887
538
553
4483
4498
GGCATCAGGACCCTTG
58
3501
|
|
1128907
591
606
4675
4690
CCATGGAGGCACGGGT
50
3502
|
|
1128927
624
639
4708
4723
AGGCGGTGGCCCTCCA
69
3503
|
|
1128947
655
670
4739
4754
GGCTCCGGTGTAGCCC
97
3504
|
|
1128967
700
715
4929
4944
CCCGCGGCCATCATAG
100
3505
|
|
1128987
752
767
4981
4996
GCTGACAGGGCGCACC
48
3506
|
|
1129007
801
816
5030
5045
CGCGCTTGCTCGGCAG
71
3507
|
|
1129027
842
857
N/A
N/A
CCGGGTTCCGGCAGAA
51
3508
|
|
1129047
884
899
5198
5213
GGTCGCGGTTCAGCAC
44
3509
|
|
1129067
976
991
5290
5305
TGGGACATGAAGCCTA
61
3510
|
|
1129087
1008
1023
5322
5337
GGCTTCGGCGGTGCCG
79
3511
|
|
1129107
1079
1094
5498
5513
GCGGCTGCTCCCGCTT
75
3512
|
|
1129127
1121
1136
5540
5555
GCCGCTGCCCGCAGCT
59
3513
|
|
1129147
1144
1159
5563
5578
CATCGAAGACAGACTC
45
3514
|
|
1129167
1202
1217
5621
5636
GCGCGGCGATGTAGGG
57
3515
|
|
1129187
1230
1245
5649
5664
CCGGCGCAGAAACTGT
73
3516
|
|
1129207
1309
1324
5969
5984
CACCGTCAGATCCTCG
52
3517
|
|
1129227
1378
1393
6038
6053
GCGGTAGGAGCGCACG
41
3518
|
|
1129247
1431
1446
N/A
N/A
CGCAACAGAGCCAGGT
48
3519
|
|
1129267
1456
1471
6199
6214
GCTGCCGTCCGCATCC
61
3520
|
|
1129287
1478
1493
6221
6236
CGTAAGGCGACAGGAG
36
3521
|
|
1129307
1525
1540
6268
6283
GGTCTCGGAGGGTCGC
69
3522
|
|
1129327
1572
1587
N/A
N/A
TCCGCCCCCTCGAACT
128
3523
|
|
1129347
1613
1628
6911
6926
GGAACGGTACCTGCGC
181
3524
|
|
1129367
1686
1701
6984
6999
AACCCTGCGCAGAGCA
17†
3525
|
|
1129387
1710
1725
7008
7023
CACGCATCGGTGCCGC
30†
3526
|
|
1129407
1781
1796
7169
7184
TGCCTTGCAGGGTGAG
23
3527
|
|
1129427
1822
1837
7210
7225
TGGCTTGTTGCGGTCA
59
3528
|
|
1129447
1853
1868
7241
7256
CCAGGTAGTAGGCCAC
56
3529
|
|
1129467
1892
1907
7280
7295
CCCTGAGCAATCAGGA
48
3530
|
|
1129487
1923
1938
7311
7326
CTGCGGAATCACCAAG
30
3531
|
|
1129507
1965
1980
7353
7368
ATGGGACACAATCTTG
70
3532
|
|
1129527
2005
2020
7393
7408
TCCTGCGCCATCCTGG
33
3533
|
|
1129547
2033
2048
7421
7436
TCAGCATTTTCAAAGC
40
3534
|
|
1129567
N/A
N/A
146
161
TGATAGCGACCCCCCA
67
3535
|
|
1129587
N/A
N/A
279
294
GTGCCCTTGTATCCAC
76
3536
|
|
1129607
N/A
N/A
423
438
AAGTCATAGCCTGCCA
115
3537
|
|
1129627
N/A
N/A
571
586
GCACCATACACATCCC
57
3538
|
|
1129647
N/A
N/A
631
646
GGGCATAAGACCTAGC
76
3539
|
|
1129667
N/A
N/A
736
751
GCACATATCTCAAGCA
53
3540
|
|
1129687
N/A
N/A
1017
1032
GAGGTTTACTGTTATC
78
3541
|
|
1129707
N/A
N/A
1207
1222
GTAGGTAATTTAGTGT
73
3542
|
|
1129727
N/A
N/A
1599
1614
CGCCATGCTTAGAGAC
82
3543
|
|
1129747
N/A
N/A
2132
2147
TGCCAAGGACCAAACC
72
3544
|
|
1129767
N/A
N/A
2292
2307
AGAACATAACCATCGC
63
3545
|
|
1129787
N/A
N/A
2520
2535
AAGGCTACCTTAGGGA
94
3546
|
|
1129807
N/A
N/A
2697
2712
AAGGTTCAACAAGGCG
61
3547
|
|
1129827
N/A
N/A
3036
3051
AGCTGGAGTAGACTCC
75
3548
|
|
1129847
N/A
N/A
3469
3484
CCCTGTACTCAACTGC
65
3549
|
|
1129867
N/A
N/A
3957
3972
CCAAAGGGTATTGTGG
74
3550
|
|
1129887
N/A
N/A
4308
4323
ACCTATTCTGTAGGCC
88
3551
|
|
1129907
N/A
N/A
4344
4359
AGTCTAGGACCATGCC
100
3552
|
|
1129927
N/A
N/A
4611
4626
AAAGACCCCCCCAGAG
116
3553
|
|
1129947
N/A
N/A
4837
4852
TGCTTTCCGCACTCTC
54
3554
|
|
1129967
N/A
N/A
5158
5173
GTCCGGGTTCCTGTAG
85
3555
|
|
1129987
N/A
N/A
5438
5453
CGCGGCCGGCTGGCCG
89
3556
|
|
1130007
N/A
N/A
5713
5728
GGGTACTCGCCGGTCC
62
3557
|
|
1130027
N/A
N/A
5862
5877
TCCCGTGTTCCAGCTT
80
3558
|
|
1130047
N/A
N/A
6164
6179
CTAACCCGGGCGGAGA
80
3559
|
|
1130067
N/A
N/A
6316
6331
GTGCCTACCCTCGAAC
112
3560
|
|
1130087
N/A
N/A
6383
6398
GTCGCGGGCTTCTTCC
54
3561
|
|
1130107
N/A
N/A
6403
6418
GCACCCGGAACGATAC
80
3562
|
|
1130127
N/A
N/A
6532
6547
AAGTTCGATTCTCCCT
70
3563
|
|
1130147
N/A
N/A
6605
6620
CACACTAGCCCGGAGC
86
3564
|
|
1130167
N/A
N/A
6670
6685
AATGGGCGGAGCGGAA
79
3565
|
|
1130187
N/A
N/A
6785
6800
GCTTGTAAACCCACTC
73
3566
|
|
1130207
N/A
N/A
6865
6880
CCCTGCGAACACAGAG
91
3567
|
|
1130227
N/A
N/A
7035
7050
CGCCAACCGGGCTAAG
98†
3568
|
|
1130247
N/A
N/A
7105
7120
ACCCTGGGTCGGAAAC
96†
3569
|
|
TABLE 58
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128747
102
117
N/A
N/A
CAAGGTGGAATCGAAA
21
3570
|
|
1128767
151
166
N/A
N/A
AACGACTGTGTGCTCT
43
3571
|
|
1128787
176
191
3527
3542
GGCAGGGCTCCCCGGT
45
3572
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
20
2426
|
|
1128808
224
239
3575
3590
TGTGGGTACATTTGTG
30
3573
|
|
1128828
288
303
3795
3810
CATCGCTGGTCCTGAT
46
3574
|
|
1128848
373
388
4181
4196
CATGTTCACACAGGTC
31
3575
|
|
1128868
468
483
4413
4428
TGGAAAAACCGGAGAA
47
3576
|
|
1128888
539
554
4484
4499
GGGCATCAGGACCCTT
69
3577
|
|
1128908
592
607
4676
4691
CCCATGGAGGCACGGG
76
3578
|
|
1128928
625
640
4709
4724
CAGGCGGTGGCCCTCC
44
3579
|
|
1128948
657
672
4741
4756
AAGGCTCCGGTGTAGC
73
3580
|
|
1128968
701
716
4930
4945
GCCCGCGGCCATCATA
52
3581
|
|
1128988
753
768
4982
4997
GGCTGACAGGGCGCAC
40
3582
|
|
1129008
802
817
5031
5046
CCGCGCTTGCTCGGCA
35
3583
|
|
1129028
843
858
N/A
N/A
TCCGGGTTCCGGCAGA
35
3584
|
|
1129048
885
900
5199
5214
CGGTCGCGGTTCAGCA
39
3585
|
|
1129068
977
992
5291
5306
GTGGGACATGAAGCCT
31
3586
|
|
1129088
1009
1024
5323
5338
AGGCTTCGGCGGTGCC
78
3587
|
|
1129108
1082
1097
5501
5516
AAGGCGGCTGCTCCCG
46
3588
|
|
1129128
1124
1139
5543
5558
GGAGCCGCTGCCCGCA
40
3589
|
|
1129148
1145
1160
5564
5579
TCATCGAAGACAGACT
36
3590
|
|
1129168
1203
1218
5622
5637
AGCGCGGCGATGTAGG
54
3591
|
|
1129188
1231
1246
5650
5665
GCCGGCGCAGAAACTG
81
3592
|
|
1129208
1311
1326
5971
5986
ACCACCGTCAGATCCT
48
3593
|
|
1129228
1379
1394
6039
6054
AGCGGTAGGAGCGCAC
48
3594
|
|
1129248
1433
1448
N/A
N/A
GGCGCAACAGAGCCAG
63
3595
|
|
1129268
1457
1472
6200
6215
AGCTGCCGTCCGCATC
42
3596
|
|
1129288
1492
1507
6235
6250
GCACACCGGCTGAACG
52
3597
|
|
1129308
1542
1557
6285
6300
GCCACCTGGCAGAGCG
73
3598
|
|
1129328
1573
1588
N/A
N/A
CTCCGCCCCCTCGAAC
135
3599
|
|
1129348
1614
1629
6912
6927
AGGAACGGTACCTGCG
127
3600
|
|
1129368
1688
1703
6986
7001
GGAACCCTGCGCAGAG
7†
3601
|
|
1129388
1711
1726
7009
7024
GCACGCATCGGTGCCG
84†
3602
|
|
1129408
1785
1800
7173
7188
ATGATGCCTTGCAGGG
16
3603
|
|
1129428
1823
1838
7211
7226
CTGGCTTGTTGCGGTC
48
3604
|
|
1129448
1854
1869
7242
7257
GCCAGGTAGTAGGCCA
64
3605
|
|
1129468
1893
1908
7281
7296
TCCCTGAGCAATCAGG
45
3606
|
|
1129488
1924
1939
7312
7327
ACTGCGGAATCACCAA
20
3607
|
|
1129508
1967
1982
7355
7370
GAATGGGACACAATCT
63
3608
|
|
1129528
2006
2021
7394
7409
TTCCTGCGCCATCCTG
35
3609
|
|
1129548
N/A
N/A
92
107
CCGAAAGTGTTGACTC
44
3610
|
|
1129568
N/A
N/A
147
162
GTGATAGCGACCCCCC
58
3611
|
|
1129588
N/A
N/A
323
338
CCCCGTTGTCTTCTTA
80
3612
|
|
1129608
N/A
N/A
425
440
ATAAGTCATAGCCTGC
93
3613
|
|
1129628
N/A
N/A
598
613
CTAGACTGCCCTGAGA
55
3614
|
|
1129648
N/A
N/A
636
651
CCCATGGGCATAAGAC
89
3615
|
|
1129668
N/A
N/A
737
752
AGCACATATCTCAAGC
75
3616
|
|
1129688
N/A
N/A
1022
1037
CCTAAGAGGTTTACTG
78
3617
|
|
1129708
N/A
N/A
1208
1223
TGTAGGTAATTTAGTG
67
3618
|
|
1129728
N/A
N/A
1695
1710
CAAGTTTGGGTAAGGC
57
3619
|
|
1129748
N/A
N/A
2137
2152
GCACTTGCCAAGGACC
42
3620
|
|
1129768
N/A
N/A
2293
2308
TAGAACATAACCATCG
59
3621
|
|
1129788
N/A
N/A
2521
2536
GAAGGCTACCTTAGGG
61
3622
|
|
1129808
N/A
N/A
2700
2715
GTTAAGGTTCAACAAG
59
3623
|
|
1129828
N/A
N/A
3037
3052
TAGCTGGAGTAGACTC
62
3624
|
|
1129848
N/A
N/A
3476
3491
GGACTTCCCCTGTACT
70
3625
|
|
1129868
N/A
N/A
3958
3973
TCCAAAGGGTATTGTG
64
3626
|
|
1129888
N/A
N/A
4309
4324
AACCTATTCTGTAGGC
74
3627
|
|
1129908
N/A
N/A
4345
4360
GAGTCTAGGACCATGC
85
3628
|
|
1129928
N/A
N/A
4613
4628
CTAAAGACCCCCCCAG
77
3629
|
|
1129948
N/A
N/A
4840
4855
ATCTGCTTTCCGCACT
23
3630
|
|
1129968
N/A
N/A
5159
5174
TGTCCGGGTTCCTGTA
59
3631
|
|
1129988
N/A
N/A
5440
5455
CCCGCGGCCGGCTGGC
91
3632
|
|
1130008
N/A
N/A
5715
5730
GCGGGTACTCGCCGGT
66
3633
|
|
1130028
N/A
N/A
5864
5879
AATCCCGTGTTCCAGC
62
3634
|
|
1130048
N/A
N/A
6165
6180
GCTAACCCGGGCGGAG
97
3635
|
|
1130068
N/A
N/A
6322
6337
GCAGTTGTGCCTACCC
53
3636
|
|
1130088
N/A
N/A
6384
6399
AGTCGCGGGCTTCTTC
78
3637
|
|
1130108
N/A
N/A
6405
6420
AGGCACCCGGAACGAT
62
3638
|
|
1130128
N/A
N/A
6533
6548
CAAGTTCGATTCTCCC
58
3639
|
|
1130148
N/A
N/A
6606
6621
CCACACTAGCCCGGAG
81
3640
|
|
1130168
N/A
N/A
6671
6686
GAATGGGCGGAGCGGA
86
3641
|
|
1130188
N/A
N/A
6786
6801
CGCTTGTAAACCCACT
58
3642
|
|
1130208
N/A
N/A
6866
6881
CCCCTGCGAACACAGA
111
3643
|
|
1130228
N/A
N/A
7036
7051
GCGCCAACCGGGCTAA
78†
3644
|
|
1130248
N/A
N/A
7106
7121
CACCCTGGGTCGGAAA
67†
3645
|
|
TABLE 59
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128748
103
118
N/A
N/A
CCAAGGTGGAATCGAA
49
3646
|
|
1128768
152
167
N/A
N/A
GAACGACTGTGTGCTC
29
3647
|
|
1128788
177
192
3528
3543
TGGCAGGGCTCCCCGG
55
3648
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
35
2426
|
|
1128809
225
240
3576
3591
TTGTGGGTACATTTGT
51
3649
|
|
1128829
290
305
3797
3812
CCCATCGCTGGTCCTG
49
3650
|
|
1128849
380
395
4188
4203
CGCTTGGCATGTTCAC
41
3651
|
|
1128869
469
484
4414
4429
GTGGAAAAACCGGAGA
58
3652
|
|
1128889
540
555
4485
4500
TGGGCATCAGGACCCT
79
3653
|
|
1128909
593
608
4677
4692
CCCCATGGAGGCACGG
52
3654
|
|
1128929
626
641
4710
4725
ACAGGCGGTGGCCCTC
48
3655
|
|
1128949
660
675
4744
4759
CAGAAGGCTCCGGTGT
79
3656
|
|
1128969
703
718
4932
4947
GAGCCCGCGGCCATCA
68
3657
|
|
1128989
754
769
4983
4998
CGGCTGACAGGGCGCA
56
3658
|
|
1129009
803
818
5032
5047
TCCGCGCTTGCTCGGC
57
3659
|
|
1129029
844
859
N/A
N/A
GTCCGGGTTCCGGCAG
51
3660
|
|
1129049
886
901
5200
5215
CCGGTCGCGGTTCAGC
55
3661
|
|
1129069
978
993
5292
5307
AGTGGGACATGAAGCC
41
3662
|
|
1129089
1010
1025
5324
5339
GAGGCTTCGGCGGTGC
59
3663
|
|
1129109
1083
1098
5502
5517
GAAGGCGGCTGCTCCC
54
3664
|
|
1129129
1125
1140
5544
5559
CGGAGCCGCTGCCCGC
70
3665
|
|
1129149
1146
1161
5565
5580
GTCATCGAAGACAGAC
58
3666
|
|
1129169
1204
1219
5623
5638
CAGCGCGGCGATGTAG
61
3667
|
|
1129189
1233
1248
5652
5667
CTGCCGGCGCAGAAAC
65
3668
|
|
1129209
1312
1327
5972
5987
CACCACCGTCAGATCC
64
3669
|
|
1129229
1380
1395
6040
6055
AAGCGGTAGGAGCGCA
66
3670
|
|
1129249
1434
1449
N/A
N/A
AGGCGCAACAGAGCCA
74
3671
|
|
1129269
1458
1473
6201
6216
CAGCTGCCGTCCGCAT
55
3672
|
|
1129289
1493
1508
6236
6251
GGCACACCGGCTGAAC
56
3673
|
|
1129309
1545
1560
6288
6303
CCGGCCACCTGGCAGA
120
3674
|
|
1129329
1575
1590
N/A
N/A
TCCTCCGCCCCCTCGA
145
3675
|
|
1129349
1615
1630
6913
6928
GAGGAACGGTACCTGC
95
3676
|
|
1129369
1689
1704
6987
7002
AGGAACCCTGCGCAGA
20†
3677
|
|
1129389
1713
1728
7011
7026
TGGCACGCATCGGTGC
65†
3678
|
|
1129409
1788
1803
7176
7191
CTGATGATGCCTTGCA
37
3679
|
|
1129429
1824
1839
7212
7227
CCTGGCTTGTTGCGGT
52
3680
|
|
1129449
1855
1870
7243
7258
GGCCAGGTAGTAGGCC
88
3681
|
|
1129469
1894
1909
7282
7297
GTCCCTGAGCAATCAG
40
3682
|
|
1129489
1925
1940
7313
7328
CACTGCGGAATCACCA
29
3683
|
|
1129509
1968
1983
7356
7371
GGAATGGGACACAATC
45
3684
|
|
1129529
2007
2022
7395
7410
GTTCCTGCGCCATCCT
37
3685
|
|
1129549
N/A
N/A
93
108
ACCGAAAGTGTTGACT
63
3686
|
|
1129569
N/A
N/A
148
163
TGTGATAGCGACCCCC
74
3687
|
|
1129589
N/A
N/A
324
339
CCCCCGTTGTCTTCTT
117
3688
|
|
1129609
N/A
N/A
426
441
TATAAGTCATAGCCTG
66
3689
|
|
1129629
N/A
N/A
600
615
GACTAGACTGCCCTGA
84
3690
|
|
1129649
N/A
N/A
637
652
GCCCATGGGCATAAGA
83
3691
|
|
1129669
N/A
N/A
744
759
CGCCAGCAGCACATAT
78
3692
|
|
1129689
N/A
N/A
1023
1038
CCCTAAGAGGTTTACT
98
3693
|
|
1129709
N/A
N/A
1214
1229
CTGGCATGTAGGTAAT
75
3694
|
|
1129729
N/A
N/A
1696
1711
CCAAGTTTGGGTAAGG
80
3695
|
|
1129749
N/A
N/A
2139
2154
GTGCACTTGCCAAGGA
80
3696
|
|
1129769
N/A
N/A
2321
2336
TGCTAGTTCAATGTTC
51
3697
|
|
1129789
N/A
N/A
2523
2538
AAGAAGGCTACCTTAG
88
3698
|
|
1129809
N/A
N/A
2703
2718
CCAGTTAAGGTTCAAC
64
3699
|
|
1129829
N/A
N/A
3038
3053
ATAGCTGGAGTAGACT
80
3700
|
|
1129849
N/A
N/A
3495
3510
GGGACAACACACTCTC
68
3701
|
|
1129869
N/A
N/A
3963
3978
GGACTTCCAAAGGGTA
66
3702
|
|
1129889
N/A
N/A
4310
4325
CAACCTATTCTGTAGG
65
3703
|
|
1129909
N/A
N/A
4347
4362
GAGAGTCTAGGACCAT
106
3704
|
|
1129929
N/A
N/A
4615
4630
GCCTAAAGACCCCCCC
88
3705
|
|
1129949
N/A
N/A
4844
4859
TCTCATCTGCTTTCCG
35
3706
|
|
1129969
N/A
N/A
5160
5175
TTGTCCGGGTTCCTGT
67
3707
|
|
1129989
N/A
N/A
5443
5458
GAGCCCGCGGCCGGCT
93
3708
|
|
1130009
N/A
N/A
5716
5731
GGCGGGTACTCGCCGG
96
3709
|
|
1130029
N/A
N/A
5865
5880
CAATCCCGTGTTCCAG
70
3710
|
|
1130049
N/A
N/A
6166
6181
AGCTAACCCGGGCGGA
105
3711
|
|
1130069
N/A
N/A
6324
6339
TAGCAGTTGTGCCTAC
70
3712
|
|
1130089
N/A
N/A
6385
6400
AAGTCGCGGGCTTCTT
93
3713
|
|
1130109
N/A
N/A
6407
6422
GTAGGCACCCGGAACG
79
3714
|
|
1130129
N/A
N/A
6534
6549
GCAAGTTCGATTCTCC
70
3715
|
|
1130149
N/A
N/A
6608
6623
TCCCACACTAGCCCGG
83
3716
|
|
1130169
N/A
N/A
6672
6687
TGAATGGGCGGAGCGG
88
3717
|
|
1130189
N/A
N/A
6787
6802
GCGCTTGTAAACCCAC
91
3718
|
|
1130209
N/A
N/A
6867
6882
GCCCCTGCGAACACAG
85
3719
|
|
1130229
N/A
N/A
7057
7072
GCCTGACGGCCTCGGG
77†
3720
|
|
1130249
N/A
N/A
7107
7122
TCACCCTGGGTCGGAA
86†
3721
|
|
TABLE 60
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128749
104
119
N/A
N/A
CCCAAGGTGGAATCGA
63
3722
|
|
1128769
153
168
N/A
N/A
AGAACGACTGTGTGCT
33
3723
|
|
1128789
178
193
3529
3544
GTGGCAGGGCTCCCCG
44
3724
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
24
2426
|
|
1128810
226
241
3577
3592
CTTGTGGGTACATTTG
40
3725
|
|
1128830
291
306
3798
3813
CCCCATCGCTGGTCCT
40
3726
|
|
1128850
381
396
4189
4204
CCGCTTGGCATGTTCA
49
3727
|
|
1128870
470
485
4415
4430
TGTGGAAAAACCGGAG
27
3728
|
|
1128890
541
556
4486
4501
GTGGGCATCAGGACCC
91
3729
|
|
1128910
594
609
4678
4693
CCCCCATGGAGGCACG
63
3730
|
|
1128930
627
642
4711
4726
CACAGGCGGTGGCCCT
53
3731
|
|
1128950
661
676
4745
4760
GCAGAAGGCTCCGGTG
118
3732
|
|
1128970
706
721
4935
4950
GCTGAGCCCGCGGCCA
61
3733
|
|
1128990
755
770
4984
4999
ACGGCTGACAGGGCGC
61
3734
|
|
1129010
804
819
5033
5048
TTCCGCGCTTGCTCGG
129
3735
|
|
1129030
845
860
N/A
N/A
TGTCCGGGTTCCGGCA
52
3736
|
|
1129050
887
902
5201
5216
GCCGGTCGCGGTTCAG
47
3737
|
|
1129070
979
994
5293
5308
GAGTGGGACATGAAGC
74
3738
|
|
1129090
1013
1028
5327
5342
GCTGAGGCTTCGGCGG
52
3739
|
|
1129110
1085
1100
5504
5519
GGGAAGGCGGCTGCTC
68
3740
|
|
1129130
1126
1141
5545
5560
GCGGAGCCGCTGCCCG
76
3741
|
|
1129150
1147
1162
5566
5581
GGTCATCGAAGACAGA
52
3742
|
|
1129170
1206
1221
5625
5640
TACAGCGCGGCGATGT
106
3743
|
|
1129190
1236
1251
5655
5670
AGGCTGCCGGCGCAGA
51
3744
|
|
1129210
1314
1329
5974
5989
AGCACCACCGTCAGAT
72
3745
|
|
1129230
1381
1396
6041
6056
CAAGCGGTAGGAGCGC
61
3746
|
|
1129250
1435
1450
N/A
N/A
AAGGCGCAACAGAGCC
60
3747
|
|
1129270
1459
1474
6202
6217
GCAGCTGCCGTCCGCA
54
3748
|
|
1129290
1494
1509
6237
6252
AGGCACACCGGCTGAA
63
3749
|
|
1129310
1546
1561
6289
6304
GCCGGCCACCTGGCAG
93
3750
|
|
1129330
1577
1592
N/A
N/A
ATTCCTCCGCCCCCTC
189
3751
|
|
1129350
1616
1631
6914
6929
AGAGGAACGGTACCTG
99
3752
|
|
1129370
1690
1705
6988
7003
GAGGAACCCTGCGCAG
17†
3753
|
|
1129390
1714
1729
7012
7027
CTGGCACGCATCGGTG
30†
3754
|
|
1129410
1789
1804
7177
7192
GCTGATGATGCCTTGC
42
3755
|
|
1129430
1825
1840
7213
7228
GCCTGGCTTGTTGCGG
60
3756
|
|
1129450
1856
1871
7244
7259
AGGCCAGGTAGTAGGC
92
3757
|
|
1129470
1895
1910
7283
7298
AGTCCCTGAGCAATCA
53
3758
|
|
1129490
1926
1941
7314
7329
TCACTGCGGAATCACC
17
3759
|
|
1129510
1984
1999
7372
7387
AGCTGGCCGCACTGGG
36
3760
|
|
1129530
2008
2023
7396
7411
AGTTCCTGCGCCATCC
19
3761
|
|
1129550
N/A
N/A
94
109
CACCGAAAGTGTTGAC
118
3762
|
|
1129570
N/A
N/A
149
164
CTGTGATAGCGACCCC
96
3763
|
|
1129590
N/A
N/A
325
340
CCCCCCGTTGTCTTCT
118
3764
|
|
1129610
N/A
N/A
427
442
CTATAAGTCATAGCCT
139
3765
|
|
1129630
N/A
N/A
602
617
TAGACTAGACTGCCCT
78
3766
|
|
1129650
N/A
N/A
644
659
CTCTAGTGCCCATGGG
75
3767
|
|
1129670
N/A
N/A
866
881
CCCATTGAAGGCCCTG
75
3768
|
|
1129690
N/A
N/A
1024
1039
ACCCTAAGAGGTTTAC
106
3769
|
|
1129710
N/A
N/A
1522
1537
TGCCACACTCTAGGTA
98
3770
|
|
1129730
N/A
N/A
1699
1714
GCACCAAGTTTGGGTA
100
3771
|
|
1129750
N/A
N/A
2141
2156
AAGTGCACTTGCCAAG
107
3772
|
|
1129770
N/A
N/A
2376
2391
GCTGTTGGTTGATGAA
85
3773
|
|
1129790
N/A
N/A
2524
2539
CAAGAAGGCTACCTTA
146
3774
|
|
1129810
N/A
N/A
2708
2723
CATTCCCAGTTAAGGT
85
3775
|
|
1129830
N/A
N/A
3039
3054
GATAGCTGGAGTAGAC
108
3776
|
|
1129850
N/A
N/A
3609
3624
TAGTCTTACCAGGGCT
82
3777
|
|
1129870
N/A
N/A
3995
4010
ATGGGACCACTCCTTC
76
3778
|
|
1129890
N/A
N/A
4311
4326
GCAACCTATTCTGTAG
109
3779
|
|
1129910
N/A
N/A
4348
4363
GGAGAGTCTAGGACCA
115
3780
|
|
1129930
N/A
N/A
4616
4631
GGCCTAAAGACCCCCC
143
3781
|
|
1129950
N/A
N/A
5067
5082
GCACCGGCAGAAGGCG
63
3782
|
|
1129970
N/A
N/A
5374
5389
TTCCTAACCTCCCGGG
97
3783
|
|
1129990
N/A
N/A
5446
5461
GGAGAGCCCGCGGCCG
95
3784
|
|
1130010
N/A
N/A
5752
5767
ACGGAGGAGCCGCGGC
62
3785
|
|
1130030
N/A
N/A
5866
5881
CCAATCCCGTGTTCCA
84
3786
|
|
1130050
N/A
N/A
6167
6182
GAGCTAACCCGGGCGG
96
3787
|
|
1130070
N/A
N/A
6327
6342
CCCTAGCAGTTGTGCC
104
3788
|
|
1130090
N/A
N/A
6386
6401
AAAGTCGCGGGCTTCT
92
3789
|
|
1130110
N/A
N/A
6409
6424
CTGTAGGCACCCGGAA
164
3790
|
|
1130130
N/A
N/A
6535
6550
AGCAAGTTCGATTCTC
80
3791
|
|
1130150
N/A
N/A
6621
6636
CGCAGAACCTGGCTCC
95
3792
|
|
1130170
N/A
N/A
6673
6688
TTGAATGGGCGGAGCG
126
3793
|
|
1130190
N/A
N/A
6809
6824
ACGATGGACAAAGCTG
147
3794
|
|
1130210
N/A
N/A
6870
6885
TCCGCCCCTGCGAACA
143
3795
|
|
1130230
N/A
N/A
7064
7079
GATTTGTGCCTGACGG
93†
3796
|
|
1130250
N/A
N/A
7109
7124
AATCACCCTGGGTCGG
115†
3797
|
|
TABLE 61
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128750
105
120
N/A
N/A
TCCCAAGGTGGAATCG
66
3798
|
|
1128770
154
169
N/A
N/A
GAGAACGACTGTGTGC
47
3799
|
|
1128790
198
213
3549
3564
CGGTGGTACTGGAAGG
45
3800
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
28
2426
|
|
1128811
228
243
3579
3594
CCCTTGTGGGTACATT
75
3801
|
|
1128831
292
307
3799
3814
TCCCCATCGCTGGTCC
70
3802
|
|
1128851
382
397
4190
4205
GCCGCTTGGCATGTTC
53
3803
|
|
1128871
472
487
4417
4432
CTTGTGGAAAAACCGG
37
3804
|
|
1128891
542
557
4487
4502
AGTGGGCATCAGGACC
64
3805
|
|
1128911
595
610
4679
4694
ACCCCCATGGAGGCAC
49
3806
|
|
1128931
628
643
4712
4727
GCACAGGCGGTGGCCC
66
3807
|
|
1128951
662
677
4746
4761
CGCAGAAGGCTCCGGT
95
3808
|
|
1128971
711
726
4940
4955
CGGTAGCTGAGCCCGC
102
3809
|
|
1128991
756
771
4985
5000
CACGGCTGACAGGGCG
53
3810
|
|
1129011
805
820
5034
5049
GTTCCGCGCTTGCTCG
43
3811
|
|
1129031
846
861
N/A
N/A
TTGTCCGGGTTCCGGC
47
3812
|
|
1129051
888
903
5202
5217
AGCCGGTCGCGGTTCA
18
3813
|
|
1129071
980
995
5294
5309
TGAGTGGGACATGAAG
50
3814
|
|
1129091
1014
1029
5328
5343
GGCTGAGGCTTCGGCG
54
3815
|
|
1129111
1086
1101
5505
5520
AGGGAAGGCGGCTGCT
44
3816
|
|
1129131
1127
1142
5546
5561
TGCGGAGCCGCTGCCC
59
3817
|
|
1129151
1150
1165
5569
5584
GCGGGTCATCGAAGAC
52
3818
|
|
1129171
1207
1222
5626
5641
GTACAGCGCGGCGATG
57
3819
|
|
1129191
1238
1253
5657
5672
TGAGGCTGCCGGCGCA
65
3820
|
|
1129211
1315
1330
5975
5990
GAGCACCACCGTCAGA
36
3821
|
|
1129231
1382
1397
6042
6057
GCAAGCGGTAGGAGCG
53
3822
|
|
1129251
1436
1451
6179
6194
GAAGGCGCAACAGAGC
60
3823
|
|
1129271
1460
1475
6203
6218
CGCAGCTGCCGTCCGC
50
3824
|
|
1129291
1495
1510
6238
6253
CAGGCACACCGGCTGA
53
3825
|
|
1129311
1554
1569
6297
6312
TGGCCCCAGCCGGCCA
78
3826
|
|
1129331
1578
1593
N/A
N/A
TATTCCTCCGCCCCCT
140
3827
|
|
1129351
1617
1632
6915
6930
GAGAGGAACGGTACCT
120
3828
|
|
1129371
1691
1706
6989
7004
CGAGGAACCCTGCGCA
41†
3829
|
|
1129391
1746
1761
7134
7149
TCCTCACACACCAGCG
15†
3830
|
|
1129411
1790
1805
7178
7193
AGCTGATGATGCCTTG
40
3831
|
|
1129431
1826
1841
7214
7229
CGCCTGGCTTGTTGCG
83
3832
|
|
1129451
1864
1879
7252
7267
CCGGATCCAGGCCAGG
66
3833
|
|
1129471
1896
1911
7284
7299
GAGTCCCTGAGCAATC
43
3834
|
|
1129491
1927
1942
7315
7330
CTCACTGCGGAATCAC
21
3835
|
|
1129511
1988
2003
7376
7391
GCGGAGCTGGCCGCAC
56
3836
|
|
1129531
2009
2024
7397
7412
GAGTTCCTGCGCCATC
18
3837
|
|
1129551
N/A
N/A
95
110
TCACCGAAAGTGTTGA
86
3838
|
|
1129571
N/A
N/A
150
165
GCTGTGATAGCGACCC
78
3839
|
|
1129591
N/A
N/A
326
341
TCCCCCCGTTGTCTTC
116
3840
|
|
1129611
N/A
N/A
428
443
ACTATAAGTCATAGCC
95
3841
|
|
1129631
N/A
N/A
603
618
CTAGACTAGACTGCCC
92
3842
|
|
1129651
N/A
N/A
645
660
ACTCTAGTGCCCATGG
86
3843
|
|
1129671
N/A
N/A
867
882
GCCCATTGAAGGCCCT
83
3844
|
|
1129691
N/A
N/A
1053
1068
ATACCTCACATGGGTT
145
3845
|
|
1129711
N/A
N/A
1555
1570
GTAAGGTCCATCTGGT
72
3846
|
|
1129731
N/A
N/A
1704
1719
ATAGGGCACCAAGTTT
93
3847
|
|
1129751
N/A
N/A
2143
2158
CAAAGTGCACTTGCCA
73
3848
|
|
1129771
N/A
N/A
2445
2460
GATGAATCAGCAACAC
56
3849
|
|
1129791
N/A
N/A
2525
2540
ACAAGAAGGCTACCTT
79
3850
|
|
1129811
N/A
N/A
2746
2761
CCATTTGTGGGCATGC
64
3851
|
|
1129831
N/A
N/A
3040
3055
AGATAGCTGGAGTAGA
87
3852
|
|
1129851
N/A
N/A
3610
3625
GTAGTCTTACCAGGGC
72
3853
|
|
1129871
N/A
N/A
3996
4011
AATGGGACCACTCCTT
60
3854
|
|
1129891
N/A
N/A
4312
4327
AGCAACCTATTCTGTA
95
3855
|
|
1129911
N/A
N/A
4349
4364
AGGAGAGTCTAGGACC
116
3856
|
|
1129931
N/A
N/A
4617
4632
GGGCCTAAAGACCCCC
94
3857
|
|
1129951
N/A
N/A
5068
5083
CGCACCGGCAGAAGGC
75
3858
|
|
1129971
N/A
N/A
5375
5390
CTTCCTAACCTCCCGG
76
3859
|
|
1129991
N/A
N/A
5448
5463
ACGGAGAGCCCGCGGC
94
3860
|
|
1130011
N/A
N/A
5753
5768
GACGGAGGAGCCGCGG
98
3861
|
|
1130031
N/A
N/A
5867
5882
CCCAATCCCGTGTTCC
76
3862
|
|
1130051
N/A
N/A
6168
6183
AGAGCTAACCCGGGCG
91
3863
|
|
1130071
N/A
N/A
6328
6343
CCCCTAGCAGTTGTGC
76
3864
|
|
1130091
N/A
N/A
6387
6402
CAAAGTCGCGGGCTTC
67
3865
|
|
1130111
N/A
N/A
6410
6425
TCTGTAGGCACCCGGA
73
3866
|
|
1130131
N/A
N/A
6536
6551
GAGCAAGTTCGATTCT
81
3867
|
|
1130151
N/A
N/A
6622
6637
TCGCAGAACCTGGCTC
76
3868
|
|
1130171
N/A
N/A
6674
6689
TTTGAATGGGCGGAGC
84
3869
|
|
1130191
N/A
N/A
6810
6825
GACGATGGACAAAGCT
82
3870
|
|
1130211
N/A
N/A
6871
6886
CTCCGCCCCTGCGAAC
122
3871
|
|
1130231
N/A
N/A
7065
7080
AGATTTGTGCCTGACG
84†
3872
|
|
1130251
N/A
N/A
7111
7126
GGAATCACCCTGGGTC
55†
3873
|
|
TABLE 62
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 3000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128751
107
122
474
489
CTTCCCAAGGTGGAAT
89
3874
|
|
1128771
155
170
N/A
N/A
TGAGAACGACTGTGTG
49
3875
|
|
1128791
199
214
3550
3565
CCGGTGGTACTGGAAG
55
3876
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
12
2426
|
|
1128812
229
244
3580
3595
GCCCTTGTGGGTACAT
56
3877
|
|
1128832
293
308
3800
3815
ATCCCCATCGCTGGTC
36
3878
|
|
1128852
383
398
4191
4206
GGCCGCTTGGCATGTT
54
3879
|
|
1128872
480
495
4425
4440
ATCTCATTCTTGTGGA
32
3880
|
|
1128892
543
558
4488
4503
CAGTGGGCATCAGGAC
48
3881
|
|
1128912
597
612
4681
4696
CGACCCCCATGGAGGC
36
3882
|
|
1128932
630
645
4714
4729
TGGCACAGGCGGTGGC
52
3883
|
|
1128952
663
678
4747
4762
TCGCAGAAGGCTCCGG
87
3884
|
|
1128972
712
727
4941
4956
GCGGTAGCTGAGCCCG
66
3885
|
|
1128992
758
773
4987
5002
CCCACGGCTGACAGGG
73
3886
|
|
1129012
806
821
5035
5050
AGTTCCGCGCTTGCTC
34
3887
|
|
1129032
862
877
5176
5191
CCACGGGCGGATGTCG
43
3888
|
|
1129052
889
904
5203
5218
CAGCCGGTCGCGGTTC
38
3889
|
|
1129072
981
996
5295
5310
ATGAGTGGGACATGAA
55
3890
|
|
1129092
1016
1031
5330
5345
TGGGCTGAGGCTTCGG
25
3891
|
|
1129112
1088
1103
5507
5522
TCAGGGAAGGCGGCTG
45
3892
|
|
1129132
1128
1143
5547
5562
TTGCGGAGCCGCTGCC
58
3893
|
|
1129152
1151
1166
5570
5585
CGCGGGTCATCGAAGA
44
3894
|
|
1129172
1208
1223
5627
5642
AGTACAGCGCGGCGAT
31
3895
|
|
1129192
1239
1254
5658
5673
ATGAGGCTGCCGGCGC
43
3896
|
|
1129212
1318
1333
5978
5993
GCCGAGCACCACCGTC
40
3897
|
|
1129232
1383
1398
6043
6058
TGCAAGCGGTAGGAGC
33
3898
|
|
1129252
1437
1452
6180
6195
TGAAGGCGCAACAGAG
44
3899
|
|
1129272
1462
1477
6205
6220
CGCGCAGCTGCCGTCC
43
3900
|
|
1129292
1496
1511
6239
6254
GCAGGCACACCGGCTG
34
3901
|
|
1129312
1556
1571
6299
6314
GGTGGCCCCAGCCGGC
60
3902
|
|
1129332
1579
1594
N/A
N/A
ATATTCCTCCGCCCCC
89
3903
|
|
1129352
1618
1633
6916
6931
GGAGAGGAACGGTACC
89
3904
|
|
1129372
1692
1707
6990
7005
TCGAGGAACCCTGCGC
46†
3905
|
|
1129392
1747
1762
7135
7150
GTCCTCACACACCAGC
5†
3906
|
|
1129412
1797
1812
7185
7200
GATCCCCAGCTGATGA
43
3907
|
|
1129432
1827
1842
7215
7230
ACGCCTGGCTTGTTGC
43
3908
|
|
1129452
1865
1880
7253
7268
CCCGGATCCAGGCCAG
41
3909
|
|
1129472
1897
1912
7285
7300
TGAGTCCCTGAGCAAT
24
3910
|
|
1129492
1928
1943
7316
7331
TCTCACTGCGGAATCA
14
3911
|
|
1129512
1989
2004
7377
7392
CGCGGAGCTGGCCGCA
39
3912
|
|
1129532
2010
2025
7398
7413
TGAGTTCCTGCGCCAT
16
3913
|
|
1129552
N/A
N/A
96
111
CTCACCGAAAGTGTTG
88
3914
|
|
1129572
N/A
N/A
186
201
GGACCCACAGGTCATG
28
3915
|
|
1129592
N/A
N/A
327
342
CTCCCCCCGTTGTCTT
85
3916
|
|
1129612
N/A
N/A
429
444
GACTATAAGTCATAGC
101
3917
|
|
1129632
N/A
N/A
604
619
ACTAGACTAGACTGCC
79
3918
|
|
1129652
N/A
N/A
649
664
GATCACTCTAGTGCCC
49
3919
|
|
1129672
N/A
N/A
889
904
GACAAGCTCTTGATCC
40
3920
|
|
1129692
N/A
N/A
1054
1069
AATACCTCACATGGGT
51
3921
|
|
1129712
N/A
N/A
1556
1571
GGTAAGGTCCATCTGG
47
3922
|
|
1129732
N/A
N/A
1708
1723
AAGGATAGGGCACCAA
53
3923
|
|
1129752
N/A
N/A
2145
2160
GGCAAAGTGCACTTGC
80
3924
|
|
1129772
N/A
N/A
2449
2464
CATTGATGAATCAGCA
53
3925
|
|
1129792
N/A
N/A
2526
2541
CACAAGAAGGCTACCT
69
3926
|
|
1129812
N/A
N/A
2806
2821
GTGCCTACTTGCTGAA
72
3927
|
|
1129832
N/A
N/A
3041
3056
CAGATAGCTGGAGTAG
39
3928
|
|
1129852
N/A
N/A
3647
3662
GTACATGTCTCCCAGG
55
3929
|
|
1129872
N/A
N/A
4014
4029
TGGTTTACCCACCTGC
59
3930
|
|
1129892
N/A
N/A
4313
4328
CAGCAACCTATTCTGT
88
3931
|
|
1129912
N/A
N/A
4350
4365
CAGGAGAGTCTAGGAC
69
3932
|
|
1129932
N/A
N/A
4618
4633
TGGGCCTAAAGACCCC
96
3933
|
|
1129952
N/A
N/A
5069
5084
GCGCACCGGCAGAAGG
101
3934
|
|
1129972
N/A
N/A
5376
5391
ACTTCCTAACCTCCCG
56
3935
|
|
1129992
N/A
N/A
5449
5464
GACGGAGAGCCCGCGG
88
3936
|
|
1130012
N/A
N/A
5754
5769
AGACGGAGGAGCCGCG
59
3937
|
|
1130032
N/A
N/A
5868
5883
CCCCAATCCCGTGTTC
82
3938
|
|
1130052
N/A
N/A
6169
6184
CAGAGCTAACCCGGGC
69
3939
|
|
1130072
N/A
N/A
6329
6344
GCCCCTAGCAGTTGTG
86
3940
|
|
1130092
N/A
N/A
6388
6403
CCAAAGTCGCGGGCTT
54
3941
|
|
1130112
N/A
N/A
6412
6427
ATTCTGTAGGCACCCG
46
3942
|
|
1130132
N/A
N/A
6537
6552
AGAGCAAGTTCGATTC
51
3943
|
|
1130152
N/A
N/A
6623
6638
GTCGCAGAACCTGGCT
60
3944
|
|
1130172
N/A
N/A
6675
6690
ATTTGAATGGGCGGAG
44
3945
|
|
1130192
N/A
N/A
6811
6826
GGACGATGGACAAAGC
66
3946
|
|
1130212
N/A
N/A
6873
6888
TCCTCCGCCCCTGCGA
152
3947
|
|
1130232
N/A
N/A
7066
7081
GAGATTTGTGCCTGAC
65†
3948
|
|
1130252
N/A
N/A
7112
7127
CGGAATCACCCTGGGT
22†
3949
|
|
TABLE 63
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7, electroporation, 2000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
124
2426
|
|
1129531
2009
2024
7397
7412
GAGTTCCTGCGCCATC
13
3837
|
|
1129948
N/A
N/A
4840
4855
ATCTGCTTTCCGCACT
134
3630
|
|
1194296
N/A
N/A
4848
4863
TCCCTCTCATCTGCTT
55
3950
|
|
1206493
N/A
N/A
4830
4845
CGCACTCTCCCTCCTC
105
3951
|
|
1208291
N/A
N/A
130
145
GAACAATCCTGGGACA
84
3952
|
|
1208323
N/A
N/A
200
215
CCTCCTAGTCACCTGG
44
3953
|
|
1208358
N/A
N/A
269
284
ATCCACCCAGTCTGGT
123
3954
|
|
1208388
N/A
N/A
315
330
TCTTCTTAAGGCCCAT
76
3955
|
|
1208422
N/A
N/A
407
422
CCGATCTGTTGCTAGT
113
3956
|
|
1208453
N/A
N/A
442
457
AAGGCAGGGAACTGAC
91
3957
|
|
1208484
N/A
N/A
531
546
AGCCAGGCCACTTACC
123
3958
|
|
1208512
N/A
N/A
599
614
ACTAGACTGCCCTGAG
102
3959
|
|
1208540
N/A
N/A
632
647
TGGGCATAAGACCTAG
113
3960
|
|
1208572
N/A
N/A
662
677
TCACACAGCTCACGAT
77
3961
|
|
1208588
N/A
N/A
734
749
ACATATCTCAAGCACC
119
3962
|
|
1208601
N/A
N/A
841
856
TGGCATGAATGATGCC
127
3963
|
|
1208614
N/A
N/A
887
902
CAAGCTCTTGATCCTT
56
3964
|
|
1208627
N/A
N/A
927
942
GTGATAAAGCTGGGCT
101
3965
|
|
1208640
N/A
N/A
969
984
TGGAGAACTCAAATCC
52
3966
|
|
1208653
N/A
N/A
1015
1030
GGTTTACTGTTATCAA
120
3967
|
|
1208666
N/A
N/A
1057
1072
CTGAATACCTCACATG
107
3968
|
|
1208679
N/A
N/A
1126
1141
TTCATCTAAAAGGTAG
73
3969
|
|
1208692
N/A
N/A
1165
1180
TATGGCAGAGCTTGAG
72
3970
|
|
1208705
N/A
N/A
1517
1532
CACTCTAGGTAAATTT
84
3971
|
|
1208718
N/A
N/A
1567
1582
AGGCCCACCAGGGTAA
111
3972
|
|
1208735
N/A
N/A
1597
1612
CCATGCTTAGAGACTC
83
3973
|
|
1208762
N/A
N/A
1690
1705
TTGGGTAAGGCAAGAG
48
3974
|
|
1208789
N/A
N/A
1718
1733
GGGATTAGGCAAGGAT
74
3975
|
|
1208813
N/A
N/A
1921
1936
CTGAGCATGAAGTTGT
80
3976
|
|
1208840
N/A
N/A
2094
2109
ATGGATCTGAGATGGA
52
3977
|
|
1208868
N/A
N/A
2126
2141
GGACCAAACCAGAACT
98
3978
|
|
1208895
N/A
N/A
2149
2164
TCAAGGCAAAGTGCAC
47
3979
|
|
1208918
N/A
N/A
2182
2197
TACCTGCTCAATAAGC
46
3980
|
|
1208944
N/A
N/A
2251
2266
CTTCAGAGGAATTGTC
64
3981
|
|
1208971
N/A
N/A
2320
2335
GCTAGTTCAATGTTCA
98
3982
|
|
1208997
N/A
N/A
2337
2352
GGAACCTAATACTGTT
73
3983
|
|
1209023
N/A
N/A
2389
2404
AACCAGATTATATGCT
80
3984
|
|
1209048
N/A
N/A
2461
2476
CCATCAGCTTAGCATT
53
3985
|
|
1209073
N/A
N/A
2493
2508
TTAGACACTTGATTCA
76
3986
|
|
1209100
N/A
N/A
2548
2563
GAAGAGCTGTGTAGTT
95
3987
|
|
1209126
N/A
N/A
2647
2662
GAATGAGTGTCCTTCT
43
3988
|
|
1209152
N/A
N/A
2707
2722
ATTCCCAGTTAAGGTT
78
3989
|
|
1209180
N/A
N/A
2750
2765
ATGGCCATTTGTGGGC
98
3990
|
|
1209206
N/A
N/A
2785
2800
TTTGTAACTCCCAAGT
80
3991
|
|
1209229
N/A
N/A
2817
2832
ACATTGAGAATGTGCC
99
3992
|
|
1209253
N/A
N/A
2896
2911
TCTGATCTGAGCCTTG
103
3993
|
|
1209277
N/A
N/A
2957
2972
CACTTGCCACAAAGTA
74
3994
|
|
1209303
N/A
N/A
3019
3034
GACTCCACAACCTGCT
130
3995
|
|
1209329
N/A
N/A
3371
3386
TAAAAGTTGGGTTCAG
78
3996
|
|
1209347
N/A
N/A
3441
3456
CCTCAGGGTCTGGTCA
117
3997
|
|
1209360
N/A
N/A
3474
3489
ACTTCCCCTGTACTCA
143
3998
|
|
1209373
N/A
N/A
3511
3526
GACAGTGAGAACTGCA
123
3999
|
|
1209386
N/A
N/A
3658
3673
GGACAGGCAGGGTACA
76
4000
|
|
1209399
N/A
N/A
3703
3718
CCCTATCACAGTCCCC
64
4001
|
|
1209412
N/A
N/A
3879
3894
AATGAGGCGGGAGGAG
122
4002
|
|
1209425
N/A
N/A
3923
3938
GTCCAGAATCCCAGGT
86
4003
|
|
1209438
N/A
N/A
3998
4013
AAAATGGGACCACTCC
144
4004
|
|
1209451
N/A
N/A
4034
4049
CTACTCCAAGTTTCCA
71
4005
|
|
1209464
N/A
N/A
4067
4082
GCCCTTCTTGAACCTA
88
4006
|
|
1209477
N/A
N/A
4269
4284
ACCCCGCCCAGGTCCT
48
4007
|
|
1209490
N/A
N/A
4331
4346
GCCAAGTCTCCGAGTA
76
4008
|
|
1209503
N/A
N/A
4364
4379
AGGGATAGTGGTCTCA
73
4009
|
|
1209516
N/A
N/A
4609
4624
AGACCCCCCCAGAGAG
71
4010
|
|
1209529
N/A
N/A
4803
4818
GCCCGTCCCACCTGGG
78
4011
|
|
1209542
N/A
N/A
5119
5134
GCGCCGGGAGCCCGGA
62
4012
|
|
1209555
N/A
N/A
5430
5445
GCTGGCCGGAATCTAG
61
4013
|
|
1209568
N/A
N/A
5745
5760
AGCCGCGGCCCCTGGG
49
4014
|
|
1209581
N/A
N/A
5860
5875
CCGTGTTCCAGCTTCC
53
4015
|
|
1209594
N/A
N/A
6095
6110
GCGCCCCCACGCACCC
74
4016
|
|
1209607
N/A
N/A
6357
6372
GTGATCAAAGGTCTCC
26
4017
|
|
1209620
N/A
N/A
6436
6451
CAACCCATCAGGTCAG
41
4018
|
|
1209633
N/A
N/A
6474
6489
CCCTGGGATTCTACCT
65
4019
|
|
1209646
N/A
N/A
6542
6557
AACCAAGAGCAAGTTC
38
4020
|
|
1209659
N/A
N/A
6625
6640
GAGTCGCAGAACCTGG
25
4021
|
|
1209672
N/A
N/A
6680
6695
CCAGGATTTGAATGGG
83
4022
|
|
1209685
N/A
N/A
6780
6795
TAAACCCACTCATGCC
44
4023
|
|
1209698
N/A
N/A
6839
6854
ACACCCCATCTGACAA
63
4024
|
|
1209711
N/A
N/A
7027
7042
GGGCTAAGAGCTCACC
88†
4025
|
|
TABLE 64
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform PS internucleoside linkages
|
(Huh7 , electroporation, 2000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
FXII
ID
|
Number
Site
Site
Site
Site
Sequence (5′ to 3′)
(% UTC)
NO
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
88
2426
|
|
1129531
2009
2024
7397
7412
GAGTTCCTGCGCCATC
7
3837
|
|
1129948
N/A
N/A
4840
4855
ATCTGCTTTCCGCACT
84
3630
|
|
1194286
N/A
N/A
4835
4850
CTTTCCGCACTCTCCC
97
4026
|
|
1194297
N/A
N/A
4849
4864
CTCCCTCTCATCTGCT
54
4027
|
|
1208292
N/A
N/A
131
146
AGAACAATCCTGGGAC
91
4028
|
|
1208327
N/A
N/A
201
216
GCCTCCTAGTCACCTG
108
4029
|
|
1208361
N/A
N/A
270
285
TATCCACCCAGTCTGG
73
4030
|
|
1208392
N/A
N/A
316
331
GTCTTCTTAAGGCCCA
79
4031
|
|
1208426
N/A
N/A
413
428
CTGCCACCGATCTGTT
57
4032
|
|
1208454
N/A
N/A
460
475
ATCTACAAGGGAGAGA
107
4033
|
|
1208487
N/A
N/A
546
561
GTTCCCGGGAGGAGGA
69
4034
|
|
1208513
N/A
N/A
601
616
AGACTAGACTGCCCTG
43
4035
|
|
1208544
N/A
N/A
633
648
ATGGGCATAAGACCTA
60
4036
|
|
1208575
N/A
N/A
664
679
GATCACACAGCTCACG
112
4037
|
|
1208589
N/A
N/A
735
750
CACATATCTCAAGCAC
65
4038
|
|
1208602
N/A
N/A
842
857
GTGGCATGAATGATGC
94
4039
|
|
1208615
N/A
N/A
888
903
ACAAGCTCTTGATCCT
96
4040
|
|
1208628
N/A
N/A
928
943
AGTGATAAAGCTGGGC
89
4041
|
|
1208641
N/A
N/A
970
985
TTGGAGAACTCAAATC
81
4042
|
|
1208654
N/A
N/A
1016
1031
AGGTTTACTGTTATCA
95
4043
|
|
1208667
N/A
N/A
1058
1073
GCTGAATACCTCACAT
74
4044
|
|
1208680
N/A
N/A
1128
1143
CCTTCATCTAAAAGGT
113
4045
|
|
1208693
N/A
N/A
1166
1181
CTATGGCAGAGCTTGA
111
4046
|
|
1208706
N/A
N/A
1518
1533
ACACTCTAGGTAAATT
83
4047
|
|
1208719
N/A
N/A
1573
1588
CAAGTCAGGCCCACCA
168
4048
|
|
1208738
N/A
N/A
1598
1613
GCCATGCTTAGAGACT
47
4049
|
|
1208764
N/A
N/A
1691
1706
TTTGGGTAAGGCAAGA
53
4050
|
|
1208792
N/A
N/A
1751
1766
TGGACGGACAGAGAGG
64
4051
|
|
1208816
N/A
N/A
1922
1937
GCTGAGCATGAAGTTG
38
4052
|
|
1208843
N/A
N/A
2111
2126
TAGGACAGTAAATGGA
64
4053
|
|
1208870
N/A
N/A
2127
2142
AGGACCAAACCAGAAC
125
4054
|
|
1208896
N/A
N/A
2151
2166
GTTCAAGGCAAAGTGC
68
4055
|
|
1208921
N/A
N/A
2183
2198
GTACCTGCTCAATAAG
102
4056
|
|
1208947
N/A
N/A
2266
2281
AATTGCCTGTGTCTTC
47
4057
|
|
1208973
N/A
N/A
2322
2337
TTGCTAGTTCAATGTT
60
4058
|
|
1209000
N/A
N/A
2338
2353
AGGAACCTAATACTGT
104
4059
|
|
1209024
N/A
N/A
2390
2405
AAACCAGATTATATGC
75
4060
|
|
1209050
N/A
N/A
2463
2478
TGCCATCAGCTTAGCA
72
4061
|
|
1209076
N/A
N/A
2495
2510
TGTTAGACACTTGATT
92
4062
|
|
1209101
N/A
N/A
2549
2564
TGAAGAGCTGTGTAGT
83
4063
|
|
1209127
N/A
N/A
2648
2663
TGAATGAGTGTCCTTC
131
4064
|
|
1209155
N/A
N/A
2709
2724
ACATTCCCAGTTAAGG
93
4065
|
|
1209183
N/A
N/A
2771
2786
GTCAATACTTGAAATG
88
4066
|
|
1209207
N/A
N/A
2786
2801
ATTTGTAACTCCCAAG
108
4067
|
|
1209232
N/A
N/A
2818
2833
TACATTGAGAATGTGC
72
4068
|
|
1209254
N/A
N/A
2901
2916
ACCACTCTGATCTGAG
70
4069
|
|
1209280
N/A
N/A
2962
2977
AAGAGCACTTGCCACA
77
4070
|
|
1209305
N/A
N/A
3028
3043
TAGACTCCTGACTCCA
141
4071
|
|
1209332
N/A
N/A
3372
3387
CTAAAAGTTGGGTTCA
77
4072
|
|
1209348
N/A
N/A
3460
3475
CAACTGCTCAGGCACT
117
4073
|
|
1209361
N/A
N/A
3485
3500
ACTCTCTGAGGACTTC
101
4074
|
|
1209374
N/A
N/A
3605
3620
CTTACCAGGGCTGAGG
93
4075
|
|
1209387
N/A
N/A
3660
3675
AAGGACAGGCAGGGTA
86
4076
|
|
1209400
N/A
N/A
3705
3720
TGCCCTATCACAGTCC
96
4077
|
|
1209413
N/A
N/A
3880
3895
TAATGAGGCGGGAGGA
106
4078
|
|
1209426
N/A
N/A
3949
3964
TATTGTGGAGGGAGAG
131
4079
|
|
1209439
N/A
N/A
3999
4014
CAAAATGGGACCACTC
58
4080
|
|
1209452
N/A
N/A
4036
4051
TGCTACTCCAAGTTTC
72
4081
|
|
1209465
N/A
N/A
4068
4083
GGCCCTTCTTGAACCT
47
4082
|
|
1209478
N/A
N/A
4271
4286
GCACCCCGCCCAGGTC
90
4083
|
|
1209491
N/A
N/A
4334
4349
CATGCCAAGTCTCCGA
73
4084
|
|
1209504
N/A
N/A
4368
4383
AAAGAGGGATAGTGGT
47
4085
|
|
1209517
N/A
N/A
4612
4627
TAAAGACCCCCCCAGA
95
4086
|
|
1209530
N/A
N/A
4804
4819
AGCCCGTCCCACCTGG
97
4087
|
|
1209543
N/A
N/A
5146
5161
GTAGCCACACGACGGG
31
4088
|
|
1209556
N/A
N/A
5431
5446
GGCTGGCCGGAATCTA
37
4089
|
|
1209569
N/A
N/A
5746
5761
GAGCCGCGGCCCCTGG
67
4090
|
|
1209582
N/A
N/A
5863
5878
ATCCCGTGTTCCAGCT
58
4091
|
|
1209595
N/A
N/A
6111
6126
TTCCCGTCCCCGCGGG
82
4092
|
|
1209608
N/A
N/A
6359
6374
CAGTGATCAAAGGTCT
34
4093
|
|
1209621
N/A
N/A
6439
6454
TCACAACCCATCAGGT
43
4094
|
|
1209634
N/A
N/A
6476
6491
GGCCCTGGGATTCTAC
89
4095
|
|
1209647
N/A
N/A
6543
6558
GAACCAAGAGCAAGTT
64
4096
|
|
1209660
N/A
N/A
6631
6646
CATCCAGAGTCGCAGA
36
4097
|
|
1209673
N/A
N/A
6681
6696
GCCAGGATTTGAATGG
84
4098
|
|
1209686
N/A
N/A
6782
6797
TGTAAACCCACTCATG
101
4099
|
|
1209699
N/A
N/A
6840
6855
CACACCCCATCTGACA
86
4100
|
|
1209712
N/A
N/A
7058
7073
TGCCTGACGGCCTCGG
47†
4101
|
|
TABLE 65
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with
|
uniform PS internucleoside linkages (Huh7,
|
electroporation, 2000 nM modified oligonucleotide)
|
SEQ ID
SEQ ID
SEQ ID
SEQ ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
Sequence
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
102
2426
|
|
1129531
2009
2024
7397
7412
GAGTTCCTGCGCCATC
25
3837
|
|
1129948
N/A
N/A
4840
4855
ATCTGCTTTCCGCACT
76
3630
|
|
1194287
N/A
N/A
4836
4851
GCTTTCCGCACTCTCC
48
4102
|
|
1206494
N/A
N/A
4854
4869
CCTGCCTCCCTCTCAT
92
4103
|
|
1208296
N/A
N/A
133
148
CCAGAACAATCCTGGG
62
4104
|
|
1208328
N/A
N/A
203
218
AGGCCTCCTAGTCACC
56
4105
|
|
1208363
N/A
N/A
274
289
CTTGTATCCACCCAGT
79
4106
|
|
1208393
N/A
N/A
321
336
CCGTTGTCTTCTTAAG
67
4107
|
|
1208427
N/A
N/A
414
429
CCTGCCACCGATCTGT
94
4108
|
|
1208458
N/A
N/A
461
476
AATCTACAAGGGAGAG
55
4109
|
|
1208489
N/A
N/A
547
562
GGTTCCCGGGAGGAGG
51
4110
|
|
1208515
N/A
N/A
605
620
CACTAGACTAGACTGC
69
4111
|
|
1208545
N/A
N/A
634
649
CATGGGCATAAGACCT
52
4112
|
|
1208577
N/A
N/A
672
687
CCCTCAAGGATCACAC
68
4113
|
|
1208590
N/A
N/A
738
753
CAGCACATATCTCAAG
64
4114
|
|
1208603
N/A
N/A
862
877
TTGAAGGCCCTGGCCT
45
4115
|
|
1208616
N/A
N/A
895
910
GCCAGAGACAAGCTCT
87
4116
|
|
1208629
N/A
N/A
932
947
CATAAGTGATAAAGCT
79
4117
|
|
1208642
N/A
N/A
975
990
GAAGCTTGGAGAACTC
88
4118
|
|
1208655
N/A
N/A
1018
1033
AGAGGTTTACTGTTAT
57
4119
|
|
1208668
N/A
N/A
1062
1077
ATGGGCTGAATACCTC
95
4120
|
|
1208681
N/A
N/A
1129
1144
CCCTTCATCTAAAAGG
133
4121
|
|
1208694
N/A
N/A
1167
1182
GCTATGGCAGAGCTTG
84
4122
|
|
1208707
N/A
N/A
1519
1534
CACACTCTAGGTAAAT
76
4123
|
|
1208720
N/A
N/A
1574
1589
CCAAGTCAGGCCCACC
74
4124
|
|
1208739
N/A
N/A
1600
1615
ACGCCATGCTTAGAGA
76
4125
|
|
1208766
N/A
N/A
1692
1707
GTTTGGGTAAGGCAAG
60
4126
|
|
1208795
N/A
N/A
1756
1771
TGGGATGGACGGACAG
83
4127
|
|
1208817
N/A
N/A
1923
1938
AGCTGAGCATGAAGTT
144
4128
|
|
1208844
N/A
N/A
2112
2127
CTAGGACAGTAAATGG
73
4129
|
|
1208873
N/A
N/A
2129
2144
CAAGGACCAAACCAGA
91
4130
|
|
1208898
N/A
N/A
2152
2167
TGTTCAAGGCAAAGTG
72
4131
|
|
1208922
N/A
N/A
2190
2205
GCACAAAGTACCTGCT
71
4132
|
|
1208948
N/A
N/A
2267
2282
GAATTGCCTGTGTCTT
72
4133
|
|
1208976
N/A
N/A
2323
2338
TTTGCTAGTTCAATGT
62
4134
|
|
1209001
N/A
N/A
2339
2354
CAGGAACCTAATACTG
61
4135
|
|
1209027
N/A
N/A
2442
2457
GAATCAGCAACACATA
85
4136
|
|
1209051
N/A
N/A
2465
2480
AGTGCCATCAGCTTAG
111
4137
|
|
1209077
N/A
N/A
2496
2511
GTGTTAGACACTTGAT
76
4138
|
|
1209104
N/A
N/A
2551
2566
GCTGAAGAGCTGTGTA
74
4139
|
|
1209130
N/A
N/A
2649
2664
TTGAATGAGTGTCCTT
102
4140
|
|
1209156
N/A
N/A
2710
2725
CACATTCCCAGTTAAG
100
4141
|
|
1209184
N/A
N/A
2774
2789
CAAGTCAATACTTGAA
50
4142
|
|
1209209
N/A
N/A
2803
2818
CCTACTTGCTGAATTT
96
4143
|
|
1209234
N/A
N/A
2819
2834
CTACATTGAGAATGTG
55
4144
|
|
1209257
N/A
N/A
2902
2917
AACCACTCTGATCTGA
68
4145
|
|
1209281
N/A
N/A
2964
2979
GGAAGAGCACTTGCCA
79
4146
|
|
1209307
N/A
N/A
3029
3044
GTAGACTCCTGACTCC
79
4147
|
|
1209333
N/A
N/A
3374
3389
CTCTAAAAGTTGGGTT
55
4148
|
|
1209349
N/A
N/A
3461
3476
TCAACTGCTCAGGCAC
73
4149
|
|
1209362
N/A
N/A
3491
3506
CAACACACTCTCTGAG
69
4150
|
|
1209375
N/A
N/A
3606
3621
TCTTACCAGGGCTGAG
90
4151
|
|
1209388
N/A
N/A
3669
3684
CTTGGACAGAAGGACA
93
4152
|
|
1209401
N/A
N/A
3706
3721
CTGCCCTATCACAGTC
65
4153
|
|
1209414
N/A
N/A
3887
3902
AGGAGAGTAATGAGGC
80
4154
|
|
1209427
N/A
N/A
3950
3965
GTATTGTGGAGGGAGA
70
4155
|
|
1209440
N/A
N/A
4003
4018
CCTGCAAAATGGGACC
69
4156
|
|
1209453
N/A
N/A
4037
4052
TTGCTACTCCAAGTTT
69
4157
|
|
1209466
N/A
N/A
4070
4085
AAGGCCCTTCTTGAAC
62
4158
|
|
1209479
N/A
N/A
4272
4287
AGCACCCCGCCCAGGT
73
4159
|
|
1209492
N/A
N/A
4335
4350
CCATGCCAAGTCTCCG
87
4160
|
|
1209505
N/A
N/A
4369
4384
CAAAGAGGGATAGTGG
92
4161
|
|
1209518
N/A
N/A
4623
4638
ACCCCTGGGCCTAAAG
91
4162
|
|
1209531
N/A
N/A
4806
4821
CAAGCCCGTCCCACCT
52
4163
|
|
1209544
N/A
N/A
5151
5166
TTCCTGTAGCCACACG
50
4164
|
|
1209557
N/A
N/A
5432
5447
CGGCTGGCCGGAATCT
46
4165
|
|
1209570
N/A
N/A
5747
5762
GGAGCCGCGGCCCCTG
62
4166
|
|
1209583
N/A
N/A
5876
5891
CTCCCGAACCCCAATC
52
4167
|
|
1209596
N/A
N/A
6114
6129
CTCTTCCCGTCCCCGC
53
4168
|
|
1209609
N/A
N/A
6362
6377
ACCCAGTGATCAAAGG
40
4169
|
|
1209622
N/A
N/A
6440
6455
CTCACAACCCATCAGG
42
4170
|
|
1209635
N/A
N/A
6477
6492
AGGCCCTGGGATTCTA
39
4171
|
|
1209648
N/A
N/A
6544
6559
GGAACCAAGAGCAAGT
40
4172
|
|
1209661
N/A
N/A
6632
6647
CCATCCAGAGTCGCAG
38
4173
|
|
1209674
N/A
N/A
6712
6727
CATAGGCAAGGAGGCT
34
4174
|
|
1209687
N/A
N/A
6799
6814
AAGCTGCTCCAGGCGC
57
4175
|
|
1209700
N/A
N/A
6849
6864
CGCCTTCTTCACACCC
117
4176
|
|
1209713
N/A
N/A
7059
7074
GTGCCTGACGGCCTCG
68†
4177
|
|
TABLE 66
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with
|
uniform PS internucleoside linkages (Huh7,
|
electroporation, 2000 nM modified oligonucleotide)
|
SEQ ID
SEQ ID
SEQ ID
SEQ ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
Sequence
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
99
2426
|
|
1129531
2009
2024
7397
7412
GAGTTCCTGCGCCATC
16
3837
|
|
1129947
N/A
N/A
4837
4852
TGCTTTCCGCACTCTC
51
3554
|
|
1129948
N/A
N/A
4840
4855
ATCTGCTTTCCGCACT
70
3630
|
|
1208266
N/A
N/A
98
113
CACTCACCGAAAGTGT
65
4178
|
|
1208297
N/A
N/A
136
151
CCCCCAGAACAATCCT
84
4179
|
|
1208332
N/A
N/A
205
220
ATAGGCCTCCTAGTCA
97
4180
|
|
1208366
N/A
N/A
275
290
CCTTGTATCCACCCAG
55
4181
|
|
1208397
N/A
N/A
322
337
CCCGTTGTCTTCTTAA
87
4182
|
|
1208431
N/A
N/A
417
432
TAGCCTGCCACCGATC
78
4183
|
|
1208459
N/A
N/A
462
477
GAATCTACAAGGGAGA
52
4184
|
|
1208492
N/A
N/A
556
571
CCACCCAAGGGTTCCC
81
4185
|
|
1208516
N/A
N/A
609
624
TAGGCACTAGACTAGA
97
4186
|
|
1208549
N/A
N/A
635
650
CCATGGGCATAAGACC
38
4187
|
|
1208578
N/A
N/A
674
689
TGCCCTCAAGGATCAC
46
4188
|
|
1208591
N/A
N/A
740
755
AGCAGCACATATCTCA
95
4189
|
|
1208604
N/A
N/A
863
878
ATTGAAGGCCCTGGCC
47
4190
|
|
1208617
N/A
N/A
896
911
TGCCAGAGACAAGCTC
43
4191
|
|
1208630
N/A
N/A
934
949
CTCATAAGTGATAAAG
58
4192
|
|
1208643
N/A
N/A
977
992
TTGAAGCTTGGAGAAC
52
4193
|
|
1208656
N/A
N/A
1019
1034
AAGAGGTTTACTGTTA
76
4194
|
|
1208669
N/A
N/A
1063
1078
GATGGGCTGAATACCT
51
4195
|
|
1208682
N/A
N/A
1130
1145
ACCCTTCATCTAAAAG
56
4196
|
|
1208695
N/A
N/A
1168
1183
AGCTATGGCAGAGCTT
98
4197
|
|
1208708
N/A
N/A
1520
1535
CCACACTCTAGGTAAA
44
4198
|
|
1208721
N/A
N/A
1575
1590
ACCAAGTCAGGCCCAC
92
4199
|
|
1208742
N/A
N/A
1620
1635
TTGGAATGTGCTGGGC
58
4200
|
|
1208768
N/A
N/A
1693
1708
AGTTTGGGTAAGGCAA
62
4201
|
|
1208796
N/A
N/A
1759
1774
AGATGGGATGGACGGA
47
4202
|
|
1208820
N/A
N/A
1924
1939
TAGCTGAGCATGAAGT
43
4203
|
|
1208847
N/A
N/A
2113
2128
ACTAGGACAGTAAATG
50
4204
|
|
1208874
N/A
N/A
2130
2145
CCAAGGACCAAACCAG
67
4205
|
|
1208899
N/A
N/A
2169
2184
AGCTTTTGCCACTATT
90
4206
|
|
1208925
N/A
N/A
2191
2206
GGCACAAAGTACCTGC
71
4207
|
|
1208951
N/A
N/A
2269
2284
GAGAATTGCCTGTGTC
92
4208
|
|
1208977
N/A
N/A
2325
2340
TGTTTGCTAGTTCAAT
27
4209
|
|
1209004
N/A
N/A
2340
2355
ACAGGAACCTAATACT
74
4210
|
|
1209028
N/A
N/A
2446
2461
TGATGAATCAGCAACA
59
4211
|
|
1209053
N/A
N/A
2467
2482
ATAGTGCCATCAGCTT
50
4212
|
|
1209080
N/A
N/A
2498
2513
GTGTGTTAGACACTTG
50
4213
|
|
1209105
N/A
N/A
2570
2585
AATGGCCTCTGAGCCT
46
4214
|
|
1209131
N/A
N/A
2650
2665
ATTGAATGAGTGTCCT
117
4215
|
|
1209159
N/A
N/A
2711
2726
GCACATTCCCAGTTAA
49
4216
|
|
1209187
N/A
N/A
2775
2790
CCAAGTCAATACTTGA
45
4217
|
|
1209210
N/A
N/A
2804
2819
GCCTACTTGCTGAATT
35
4218
|
|
1209236
N/A
N/A
2824
2839
TGGTTCTACATTGAGA
93
4219
|
|
1209258
N/A
N/A
2903
2918
TAACCACTCTGATCTG
49
4220
|
|
1209284
N/A
N/A
2965
2980
AGGAAGAGCACTTGCC
55
4221
|
|
1209309
N/A
N/A
3030
3045
AGTAGACTCCTGACTC
49
4222
|
|
1209336
N/A
N/A
3377
3392
CTGCTCTAAAAGTTGG
64
4223
|
|
1209350
N/A
N/A
3462
3477
CTCAACTGCTCAGGCA
61
4224
|
|
1209363
N/A
N/A
3492
3507
ACAACACACTCTCTGA
48
4225
|
|
1209376
N/A
N/A
3607
3622
GTCTTACCAGGGCTGA
59
4226
|
|
1209389
N/A
N/A
3670
3685
CCTTGGACAGAAGGAC
49
4227
|
|
1209402
N/A
N/A
3707
3722
CCTGCCCTATCACAGT
92
4228
|
|
1209415
N/A
N/A
3889
3904
CCAGGAGAGTAATGAG
49
4229
|
|
1209428
N/A
N/A
3951
3966
GGTATTGTGGAGGGAG
56
4230
|
|
1209441
N/A
N/A
4012
4027
GTTTACCCACCTGCAA
39
4231
|
|
1209454
N/A
N/A
4038
4053
CTTGCTACTCCAAGTT
108
4232
|
|
1209467
N/A
N/A
4073
4088
GCCAAGGCCCTTCTTG
56
4233
|
|
1209480
N/A
N/A
4295
4310
GCCCAGGGTTGCCCCT
65
4234
|
|
1209493
N/A
N/A
4336
4351
ACCATGCCAAGTCTCC
46
4235
|
|
1209506
N/A
N/A
4370
4385
ACAAAGAGGGATAGTG
67
4236
|
|
1209519
N/A
N/A
4625
4640
CCACCCCTGGGCCTAA
134
4237
|
|
1209532
N/A
N/A
4807
4822
GCAAGCCCGTCCCACC
81
4238
|
|
1209545
N/A
N/A
5152
5167
GTTCCTGTAGCCACAC
23
4239
|
|
1209558
N/A
N/A
5435
5450
GGCCGGCTGGCCGGAA
84
4240
|
|
1209571
N/A
N/A
5748
5763
AGGAGCCGCGGCCCCT
64
4241
|
|
1209584
N/A
N/A
5878
5893
TGCTCCCGAACCCCAA
35
4242
|
|
1209597
N/A
N/A
6120
6135
CAAGCTCTCTTCCCGT
53
4243
|
|
1209610
N/A
N/A
6365
6380
CTAACCCAGTGATCAA
29
4244
|
|
1209623
N/A
N/A
6443
6458
ATTCTCACAACCCATC
64
4245
|
|
1209636
N/A
N/A
6486
6501
GTGAATCCCAGGCCCT
41
4246
|
|
1209649
N/A
N/A
6553
6568
CGCCCAGAGGGAACCA
116
4247
|
|
1209662
N/A
N/A
6633
6648
CCCATCCAGAGTCGCA
30
4248
|
|
1209675
N/A
N/A
6713
6728
TCATAGGCAAGGAGGC
36
4249
|
|
1209688
N/A
N/A
6800
6815
AAAGCTGCTCCAGGCG
27
4250
|
|
1209701
N/A
N/A
6850
6865
GCGCCTTCTTCACACC
85
4251
|
|
1209714
N/A
N/A
7063
7078
ATTTGTGCCTGACGGC
53†
4252
|
|
TABLE 67
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with
|
uniform PS internucleoside linkages (Huh7,
|
electroporation, 2000 nM modified oligonucleotide)
|
SEQ ID
SEQ ID
SEQ ID
SEQ ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
Sequence
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
29
2426
|
|
1129531
2009
2024
7397
7412
GAGTTCCTGCGCCATC
33
3837
|
|
1129948
N/A
N/A
4840
4855
ATCTGCTTTCCGCACT
52
3630
|
|
1194288
N/A
N/A
4838
4853
CTGCTTTCCGCACTCT
27
4253
|
|
1208267
N/A
N/A
104
119
CCACAGCACTCACCGA
128
4254
|
|
1208301
N/A
N/A
137
152
CCCCCCAGAACAATCC
177
4255
|
|
1208333
N/A
N/A
213
228
CTTTCCACATAGGCCT
68
4256
|
|
1208370
N/A
N/A
276
291
CCCTTGTATCCACCCA
41
4257
|
|
1208398
N/A
N/A
331
346
CTACCTCCCCCCGTTG
91
4258
|
|
1208432
N/A
N/A
418
433
ATAGCCTGCCACCGAT
116
4259
|
|
1208463
N/A
N/A
465
480
GTGGAATCTACAAGGG
34
4260
|
|
1208494
N/A
N/A
557
572
CCCACCCAAGGGTTCC
44
4261
|
|
1208519
N/A
N/A
616
631
CACCAGGTAGGCACTA
93
4262
|
|
1208550
N/A
N/A
638
653
TGCCCATGGGCATAAG
43
4263
|
|
1208579
N/A
N/A
685
700
AGCCCATACCCTGCCC
77
4264
|
|
1208592
N/A
N/A
741
756
CAGCAGCACATATCTC
64
4265
|
|
1208605
N/A
N/A
864
879
CATTGAAGGCCCTGGC
90
4266
|
|
1208618
N/A
N/A
897
912
ATGCCAGAGACAAGCT
56
4267
|
|
1208631
N/A
N/A
935
950
GCTCATAAGTGATAAA
54
4268
|
|
1208644
N/A
N/A
1002
1017
CAACCTGGTTTCACAG
32
4269
|
|
1208657
N/A
N/A
1020
1035
TAAGAGGTTTACTGTT
44
4270
|
|
1208670
N/A
N/A
1066
1081
GGTGATGGGCTGAATA
52
4271
|
|
1208683
N/A
N/A
1131
1146
TACCCTTCATCTAAAA
80
4272
|
|
1208696
N/A
N/A
1169
1184
GAGCTATGGCAGAGCT
67
4273
|
|
1208709
N/A
N/A
1529
1544
TGCTATGTGCCACACT
61
4274
|
|
1208722
N/A
N/A
1578
1593
CCCACCAAGTCAGGCC
56
4275
|
|
1208744
N/A
N/A
1621
1636
GTTGGAATGTGCTGGG
42
4276
|
|
1208771
N/A
N/A
1694
1709
AAGTTTGGGTAAGGCA
81
4277
|
|
1208799
N/A
N/A
1760
1775
CAGATGGGATGGACGG
57
4278
|
|
1208821
N/A
N/A
2075
2090
TTAAGATGCAAATGGG
31
4279
|
|
1208848
N/A
N/A
2115
2130
GAACTAGGACAGTAAA
40
4280
|
|
1208877
N/A
N/A
2131
2146
GCCAAGGACCAAACCA
34
4281
|
|
1208901
N/A
N/A
2170
2185
AAGCTTTTGCCACTAT
49
4282
|
|
1208926
N/A
N/A
2192
2207
TGGCACAAAGTACCTG
73
4283
|
|
1208952
N/A
N/A
2270
2285
TGAGAATTGCCTGTGT
59
4284
|
|
1208980
N/A
N/A
2326
2341
CTGTTTGCTAGTTCAA
47
4285
|
|
1209005
N/A
N/A
2342
2357
TCACAGGAACCTAATA
101
4286
|
|
1209031
N/A
N/A
2447
2462
TTGATGAATCAGCAAC
39
4287
|
|
1209054
N/A
N/A
2471
2486
TGCTATAGTGCCATCA
49
4288
|
|
1209081
N/A
N/A
2517
2532
GCTACCTTAGGGAGAA
48
4289
|
|
1209108
N/A
N/A
2572
2587
GAAATGGCCTCTGAGC
30
4290
|
|
1209134
N/A
N/A
2657
2672
CTCTCCTATTGAATGA
18
4291
|
|
1209160
N/A
N/A
2713
2728
TTGCACATTCCCAGTT
34
4292
|
|
1209188
N/A
N/A
2776
2791
CCCAAGTCAATACTTG
43
4293
|
|
1209212
N/A
N/A
2805
2820
TGCCTACTTGCTGAAT
87
4294
|
|
1209238
N/A
N/A
2825
2840
CTGGTTCTACATTGAG
20
4295
|
|
1209261
N/A
N/A
2905
2920
GTTAACCACTCTGATC
79
4296
|
|
1209285
N/A
N/A
2985
3000
AAGTCTTCAGTCCCAA
47
4297
|
|
1209312
N/A
N/A
3033
3048
TGGAGTAGACTCCTGA
72
4298
|
|
1209337
N/A
N/A
3413
3428
ACCCTCAAAAAGGTCG
45
4299
|
|
1209351
N/A
N/A
3463
3478
ACTCAACTGCTCAGGC
87
4300
|
|
1209364
N/A
N/A
3493
3508
GACAACACACTCTCTG
44
4301
|
|
1209377
N/A
N/A
3608
3623
AGTCTTACCAGGGCTG
88
4302
|
|
1209390
N/A
N/A
3673
3688
GTTCCTTGGACAGAAG
96
4303
|
|
1209403
N/A
N/A
3709
3724
ACCCTGCCCTATCACA
29
4304
|
|
1209416
N/A
N/A
3890
3905
ACCAGGAGAGTAATGA
31
4305
|
|
1209429
N/A
N/A
3954
3969
AAGGGTATTGTGGAGG
45
4306
|
|
1209442
N/A
N/A
4016
4031
CTTGGTTTACCCACCT
49
4307
|
|
1209455
N/A
N/A
4041
4056
GACCTTGCTACTCCAA
21
4308
|
|
1209468
N/A
N/A
4074
4089
GGCCAAGGCCCTTCTT
128
4309
|
|
1209481
N/A
N/A
4298
4313
TAGGCCCAGGGTTGCC
135
4310
|
|
1209494
N/A
N/A
4337
4352
GACCATGCCAAGTCTC
40
4311
|
|
1209507
N/A
N/A
4528
4543
GTTCCCAACCATCTGC
42
4312
|
|
1209520
N/A
N/A
4631
4646
AGTGAGCCACCCCTGG
91
4313
|
|
1209533
N/A
N/A
4813
4828
TTCCTGGCAAGCCCGT
60
4314
|
|
1209546
N/A
N/A
5156
5171
CCGGGTTCCTGTAGCC
58
4315
|
|
1209559
N/A
N/A
5437
5452
GCGGCCGGCTGGCCGG
56
4316
|
|
1209572
N/A
N/A
5749
5764
GAGGAGCCGCGGCCCC
40
4317
|
|
1209585
N/A
N/A
5879
5894
CTGCTCCCGAACCCCA
48
4318
|
|
1209598
N/A
N/A
6121
6136
CCAAGCTCTCTTCCCG
61
4319
|
|
1209611
N/A
N/A
6366
6381
CCTAACCCAGTGATCA
48
4320
|
|
1209624
N/A
N/A
6453
6468
TCACCTACACATTCTC
37
4321
|
|
1209637
N/A
N/A
6487
6502
AGTGAATCCCAGGCCC
138
4322
|
|
1209650
N/A
N/A
6554
6569
GCGCCCAGAGGGAACC
50
4323
|
|
1209663
N/A
N/A
6635
6650
CACCCATCCAGAGTCG
51
4324
|
|
1209676
N/A
N/A
6714
6729
TTCATAGGCAAGGAGG
45
4325
|
|
1209689
N/A
N/A
6803
6818
GACAAAGCTGCTCCAG
99
4326
|
|
1209702
N/A
N/A
6864
6879
CCTGCGAACACAGAGC
43
4327
|
|
1209715
N/A
N/A
7067
7082
TGAGATTTGTGCCTGA
44†
4328
|
|
TABLE 68
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with
|
uniform PS internucleoside linkages (Huh7,
|
electroporation, 2000 nM modified oligonucleotide)
|
SEQ ID
SEQ ID
SEQ ID
SEQ ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
Sequence
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
37
2426
|
|
1129531
2009
2024
7397
7412
GAGTTCCTGCGCCATC
25
3837
|
|
1129948
N/A
N/A
4840
4855
ATCTGCTTTCCGCACT
44
3630
|
|
1194289
N/A
N/A
4839
4854
TCTGCTTTCCGCACTC
28
4329
|
|
1208271
N/A
N/A
116
131
CAATCCTGGTTCCCAC
68
4330
|
|
1208302
N/A
N/A
154
169
CATGGCTGTGATAGCG
73
4331
|
|
1208337
N/A
N/A
215
230
ACCTTTCCACATAGGC
81
4332
|
|
1208371
N/A
N/A
278
293
TGCCCTTGTATCCACC
43
4333
|
|
1208402
N/A
N/A
342
357
CCAAACCCTTTCTACC
59
4334
|
|
1208435
N/A
N/A
419
434
CATAGCCTGCCACCGA
29
4335
|
|
1208464
N/A
N/A
467
482
AGGTGGAATCTACAAG
38
4336
|
|
1208497
N/A
N/A
558
573
CCCCACCCAAGGGTTC
68
4337
|
|
1208520
N/A
N/A
617
632
GCACCAGGTAGGCACT
44
4338
|
|
1208554
N/A
N/A
646
661
CACTCTAGTGCCCATG
111
4339
|
|
1208580
N/A
N/A
695
710
ACTTAGACACAGCCCA
43
4340
|
|
1208593
N/A
N/A
745
760
GCGCCAGCAGCACATA
55
4341
|
|
1208606
N/A
N/A
865
880
CCATTGAAGGCCCTGG
39
4342
|
|
1208619
N/A
N/A
898
913
GATGCCAGAGACAAGC
26
4343
|
|
1208632
N/A
N/A
936
951
AGCTCATAAGTGATAA
73
4344
|
|
1208645
N/A
N/A
1005
1020
TATCAACCTGGTTTCA
47
4345
|
|
1208658
N/A
N/A
1021
1036
CTAAGAGGTTTACTGT
35
4346
|
|
1208671
N/A
N/A
1072
1087
GCACCAGGTGATGGGC
40
4347
|
|
1208684
N/A
N/A
1133
1148
GGTACCCTTCATCTAA
121
4348
|
|
1208697
N/A
N/A
1200
1215
ATTTAGTGTCTGGAAG
41
4349
|
|
1208710
N/A
N/A
1541
1556
GTTCACAGGCCCTGCT
55
4350
|
|
1208723
N/A
N/A
1579
1594
CCCCACCAAGTCAGGC
95
4351
|
|
1208745
N/A
N/A
1624
1639
AGGGTTGGAATGTGCT
44
4352
|
|
1208773
N/A
N/A
1697
1712
ACCAAGTTTGGGTAAG
39
4353
|
|
1208800
N/A
N/A
1762
1777
TGCAGATGGGATGGAC
51
4354
|
|
1208824
N/A
N/A
2080
2095
GACCTTTAAGATGCAA
40
4355
|
|
1208851
N/A
N/A
2116
2131
AGAACTAGGACAGTAA
33
4356
|
|
1208878
N/A
N/A
2133
2148
TTGCCAAGGACCAAAC
36
4357
|
|
1208902
N/A
N/A
2171
2186
TAAGCTTTTGCCACTA
29
4358
|
|
1208929
N/A
N/A
2193
2208
CTGGCACAAAGTACCT
58
4359
|
|
1208955
N/A
N/A
2271
2286
ATGAGAATTGCCTGTG
29
4360
|
|
1208981
N/A
N/A
2327
2342
ACTGTTTGCTAGTTCA
44
4361
|
|
1209008
N/A
N/A
2375
2390
CTGTTGGTTGATGAAA
83
4362
|
|
1209032
N/A
N/A
2448
2463
ATTGATGAATCAGCAA
68
4363
|
|
1209057
N/A
N/A
2472
2487
GTGCTATAGTGCCATC
25
4364
|
|
1209084
N/A
N/A
2518
2533
GGCTACCTTAGGGAGA
57
4365
|
|
1209109
N/A
N/A
2579
2594
GCTTTTGGAAATGGCC
27
4366
|
|
1209135
N/A
N/A
2673
2688
CTGCTGCTTGTTTCAG
82
4367
|
|
1209163
N/A
N/A
2716
2731
AATTTGCACATTCCCA
71
4368
|
|
1209191
N/A
N/A
2777
2792
TCCCAAGTCAATACTT
77
4369
|
|
1209213
N/A
N/A
2808
2823
ATGTGCCTACTTGCTG
58
4370
|
|
1209240
N/A
N/A
2827
2842
CTCTGGTTCTACATTG
38
4371
|
|
1209262
N/A
N/A
2906
2921
TGTTAACCACTCTGAT
106
4372
|
|
1209288
N/A
N/A
2986
3001
TAAGTCTTCAGTCCCA
33
4373
|
|
1209313
N/A
N/A
3042
3057
TCAGATAGCTGGAGTA
82
4374
|
|
1209339
N/A
N/A
3414
3429
GACCCTCAAAAAGGTC
76
4375
|
|
1209352
N/A
N/A
3464
3479
TACTCAACTGCTCAGG
47
4376
|
|
1209365
N/A
N/A
3494
3509
GGACAACACACTCTCT
73
4377
|
|
1209378
N/A
N/A
3611
3626
CGTAGTCTTACCAGGG
27
4378
|
|
1209391
N/A
N/A
3678
3693
GCAGAGTTCCTTGGAC
22
4379
|
|
1209404
N/A
N/A
3713
3728
GCCCACCCTGCCCTAT
83
4380
|
|
1209417
N/A
N/A
3891
3906
TACCAGGAGAGTAATG
34
4381
|
|
1209430
N/A
N/A
3955
3970
AAAGGGTATTGTGGAG
19
4382
|
|
1209443
N/A
N/A
4020
4035
CAAGCTTGGTTTACCC
48
4383
|
|
1209456
N/A
N/A
4053
4068
TACTTGCCTTGTGACC
42
4384
|
|
1209469
N/A
N/A
4075
4090
GGGCCAAGGCCCTTCT
51
4385
|
|
1209482
N/A
N/A
4299
4314
GTAGGCCCAGGGTTGC
33
4386
|
|
1209495
N/A
N/A
4340
4355
TAGGACCATGCCAAGT
39
4387
|
|
1209508
N/A
N/A
4530
4545
CCGTTCCCAACCATCT
60
4388
|
|
1209521
N/A
N/A
4638
4653
GGAACGCAGTGAGCCA
73
4389
|
|
1209534
N/A
N/A
4814
4829
CTTCCTGGCAAGCCCG
45
4390
|
|
1209547
N/A
N/A
5161
5176
GTTGTCCGGGTTCCTG
39
4391
|
|
1209560
N/A
N/A
5439
5454
CCGCGGCCGGCTGGCC
42
4392
|
|
1209573
N/A
N/A
5756
5771
GGAGACGGAGGAGCCG
71
4393
|
|
1209586
N/A
N/A
5885
5900
AGCCCCCTGCTCCCGA
50
4394
|
|
1209599
N/A
N/A
6126
6141
GGGCCCCAAGCTCTCT
30
4395
|
|
1209612
N/A
N/A
6372
6387
CTTCCGCCTAACCCAG
36
4396
|
|
1209625
N/A
N/A
6456
6471
GATTCACCTACACATT
40
4397
|
|
1209638
N/A
N/A
6490
6505
AGCAGTGAATCCCAGG
35
4398
|
|
1209651
N/A
N/A
6569
6584
GGCCTTTGCAGCCCGG
58
4399
|
|
1209664
N/A
N/A
6636
6651
CCACCCATCCAGAGTC
73
4400
|
|
1209677
N/A
N/A
6730
6745
GTGCCATTAATTCAAT
61
4401
|
|
1209690
N/A
N/A
6804
6819
GGACAAAGCTGCTCCA
75
4402
|
|
1209703
N/A
N/A
6868
6883
CGCCCCTGCGAACACA
27
4403
|
|
1209716
N/A
N/A
7068
7083
CTGAGATTTGTGCCTG
25†
4404
|
|
TABLE 69
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with
|
uniform PS internucleoside linkages (Huh7,
|
electroporation, 2000 nM modified oligonucleotide)
|
SEQ ID
SEQ ID
SEQ ID
SEQ ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
Sequence
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
98
2426
|
|
1129531
2009
2024
7397
7412
GAGTTCCTGCGCCATC
13
3837
|
|
1129948
N/A
N/A
4840
4855
ATCTGCTTTCCGCACT
71
3630
|
|
1194290
N/A
N/A
4841
4856
CATCTGCTTTCCGCAC
44
4405
|
|
1208272
N/A
N/A
117
132
ACAATCCTGGTTCCCA
40
4406
|
|
1208307
N/A
N/A
177
192
GGTCATGAGCAGAGGC
58
4407
|
|
1208338
N/A
N/A
217
232
TCACCTTTCCACATAG
59
4408
|
|
1208375
N/A
N/A
301
316
ATCTCCCCCAGAAATG
40
4409
|
|
1208403
N/A
N/A
343
358
CCCAAACCCTTTCTAC
86
4410
|
|
1208436
N/A
N/A
421
436
GTCATAGCCTGCCACC
72
4411
|
|
1208468
N/A
N/A
468
483
AAGGTGGAATCTACAA
47
4412
|
|
1208498
N/A
N/A
560
575
ATCCCCACCCAAGGGT
53
4413
|
|
1208524
N/A
N/A
618
633
AGCACCAGGTAGGCAC
49
4414
|
|
1208555
N/A
N/A
648
663
ATCACTCTAGTGCCCA
111
4415
|
|
1208581
N/A
N/A
696
711
CACTTAGACACAGCCC
83
4416
|
|
1208594
N/A
N/A
834
849
AATGATGCCCATGAGA
40
4417
|
|
1208607
N/A
N/A
868
883
TGCCCATTGAAGGCCC
60
4418
|
|
1208620
N/A
N/A
906
921
GGCATTCAGATGCCAG
108
4419
|
|
1208633
N/A
N/A
937
952
CAGCTCATAAGTGATA
55
4420
|
|
1208646
N/A
N/A
1006
1021
TTATCAACCTGGTTTC
57
4421
|
|
1208659
N/A
N/A
1025
1040
AACCCTAAGAGGTTTA
40
4422
|
|
1208672
N/A
N/A
1073
1088
TGCACCAGGTGATGGG
53
4423
|
|
1208685
N/A
N/A
1135
1150
CAGGTACCCTTCATCT
51
4424
|
|
1208698
N/A
N/A
1202
1217
TAATTTAGTGTCTGGA
89
4425
|
|
1208711
N/A
N/A
1547
1562
CATCTGGTTCACAGGC
53
4426
|
|
1208724
N/A
N/A
1580
1595
ACCCCACCAAGTCAGG
106
4427
|
|
1208748
N/A
N/A
1625
1640
CAGGGTTGGAATGTGC
88
4428
|
|
1208775
N/A
N/A
1698
1713
CACCAAGTTTGGGTAA
93
4429
|
|
1208802
N/A
N/A
1763
1778
ATGCAGATGGGATGGA
41
4430
|
|
1208825
N/A
N/A
2081
2096
GGACCTTTAAGATGCA
44
4431
|
|
1208852
N/A
N/A
2117
2132
CAGAACTAGGACAGTA
90
4432
|
|
1208881
N/A
N/A
2134
2149
CTTGCCAAGGACCAAA
84
4433
|
|
1208905
N/A
N/A
2172
2187
ATAAGCTTTTGCCACT
66
4434
|
|
1208930
N/A
N/A
2207
2222
ATGCTGAGCAGTGTCT
30
4435
|
|
1208956
N/A
N/A
2277
2292
CGAATAATGAGAATTG
73
4436
|
|
1208984
N/A
N/A
2328
2343
TACTGTTTGCTAGTTC
81
4437
|
|
1209009
N/A
N/A
2378
2393
ATGCTGTTGGTTGATG
39
4438
|
|
1209035
N/A
N/A
2451
2466
AGCATTGATGAATCAG
117
4439
|
|
1209058
N/A
N/A
2473
2488
TGTGCTATAGTGCCAT
111
4440
|
|
1209085
N/A
N/A
2522
2537
AGAAGGCTACCTTAGG
79
4441
|
|
1209111
N/A
N/A
2580
2595
GGCTTTTGGAAATGGC
62
4442
|
|
1209137
N/A
N/A
2698
2713
TAAGGTTCAACAAGGC
44
4443
|
|
1209166
N/A
N/A
2717
2732
AAATTTGCACATTCCC
86
4444
|
|
1209192
N/A
N/A
2778
2793
CTCCCAAGTCAATACT
105
4445
|
|
1209215
N/A
N/A
2810
2825
GAATGTGCCTACTTGC
42
4446
|
|
1209241
N/A
N/A
2851
2866
ATAGTACAGTTGATCC
40
4447
|
|
1209265
N/A
N/A
2908
2923
ACTGTTAACCACTCTG
57
4448
|
|
1209289
N/A
N/A
2988
3003
GGTAAGTCTTCAGTCC
46
4449
|
|
1209316
N/A
N/A
3043
3058
GTCAGATAGCTGGAGT
94
4450
|
|
1209340
N/A
N/A
3415
3430
AGACCCTCAAAAAGGT
40
4451
|
|
1209353
N/A
N/A
3465
3480
GTACTCAACTGCTCAG
57
4452
|
|
1209366
N/A
N/A
3496
3511
AGGGACAACACACTCT
35
4453
|
|
1209379
N/A
N/A
3625
3640
CTCCAACTCCTCTGCG
61
4454
|
|
1209392
N/A
N/A
3681
3696
CAAGCAGAGTTCCTTG
62
4455
|
|
1209405
N/A
N/A
3745
3760
GACAAGGCTTCCCTGC
106
4456
|
|
1209418
N/A
N/A
3892
3907
ATACCAGGAGAGTAAT
51
4457
|
|
1209431
N/A
N/A
3959
3974
TTCCAAAGGGTATTGT
41
4458
|
|
1209444
N/A
N/A
4021
4036
CCAAGCTTGGTTTACC
90
4459
|
|
1209457
N/A
N/A
4054
4069
CTACTTGCCTTGTGAC
42
4460
|
|
1209470
N/A
N/A
4126
4141
GAGAGATGGACATGGT
44
4461
|
|
1209483
N/A
N/A
4301
4316
CTGTAGGCCCAGGGTT
44
4462
|
|
1209496
N/A
N/A
4341
4356
CTAGGACCATGCCAAG
84
4463
|
|
1209509
N/A
N/A
4531
4546
CCCGTTCCCAACCATC
114
4464
|
|
1209522
N/A
N/A
4640
4655
AGGGAACGCAGTGAGC
74
4465
|
|
1209535
N/A
N/A
4833
4848
TTCCGCACTCTCCCTC
81
4466
|
|
1209548
N/A
N/A
5162
5177
CGTTGTCCGGGTTCCT
11
4467
|
|
1209561
N/A
N/A
5441
5456
GCCCGCGGCCGGCTGG
39
4468
|
|
1209574
N/A
N/A
5757
5772
GGGAGACGGAGGAGCC
76
4469
|
|
1209587
N/A
N/A
5886
5901
AAGCCCCCTGCTCCCG
99
4470
|
|
1209600
N/A
N/A
6173
6188
GCAACAGAGCTAACCC
52
4471
|
|
1209613
N/A
N/A
6374
6389
TTCTTCCGCCTAACCC
31
4472
|
|
1209626
N/A
N/A
6460
6475
CTGGGATTCACCTACA
58
4473
|
|
1209639
N/A
N/A
6491
6506
CAGCAGTGAATCCCAG
66
4474
|
|
1209652
N/A
N/A
6574
6589
TAGTTGGCCTTTGCAG
33
4475
|
|
1209665
N/A
N/A
6637
6652
ACCACCCATCCAGAGT
54
4476
|
|
1209678
N/A
N/A
6736
6751
GAGGAGGTGCCATTAA
37
4477
|
|
1209691
N/A
N/A
6805
6820
TGGACAAAGCTGCTCC
57
4478
|
|
1209704
N/A
N/A
6869
6884
CCGCCCCTGCGAACAC
48
4479
|
|
1209717
N/A
N/A
7069
7084
CCTGAGATTTGTGCCT
88†
4480
|
|
TABLE 70
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with uniform
|
PS internucleoside linkages (Huh7, electroporation,
|
2000 nM modified oligonucleotide)
|
SEQ ID
SEQ ID
SEQ ID
SEQ ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
Sequence
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
99
2426
|
|
1129531
2009
2024
7397
7412
GAGTTCCTGCGCCATC
11
3837
|
|
1129948
N/A
N/A
4840
4855
ATCTGCTTTCCGCACT
43
3630
|
|
1194291
N/A
N/A
4842
4857
TCATCTGCTTTCCGCA
55
4481
|
|
1208276
N/A
N/A
118
133
GACAATCCTGGTTCCC
81
4482
|
|
1208310
N/A
N/A
182
197
CCACAGGTCATGAGCA
44
4483
|
|
1208342
N/A
N/A
231
246
CTTCCGGGCTGGCCTC
104
4484
|
|
1208376
N/A
N/A
309
324
TAAGGCCCATCTCCCC
117
4485
|
|
1208407
N/A
N/A
349
364
CCAAGACCCAAACCCT
60
4486
|
|
1208438
N/A
N/A
424
439
TAAGTCATAGCCTGCC
71
4487
|
|
1208469
N/A
N/A
471
486
CCCAAGGTGGAATCTA
92
4488
|
|
1208500
N/A
N/A
570
585
CACCATACACATCCCC
44
4489
|
|
1208525
N/A
N/A
620
635
CTAGCACCAGGTAGGC
52
4490
|
|
1208559
N/A
N/A
650
665
CGATCACTCTAGTGCC
69
4491
|
|
1208582
N/A
N/A
698
713
GGCACTTAGACACAGC
61
4492
|
|
1208595
N/A
N/A
835
850
GAATGATGCCCATGAG
65
4493
|
|
1208608
N/A
N/A
869
884
TTGCCCATTGAAGGCC
61
4494
|
|
1208621
N/A
N/A
907
922
AGGCATTCAGATGCCA
43
4495
|
|
1208634
N/A
N/A
951
966
CGCCCAGAGTCACCCA
58
4496
|
|
1208647
N/A
N/A
1007
1022
GTTATCAACCTGGTTT
76
4497
|
|
1208660
N/A
N/A
1026
1041
CAACCCTAAGAGGTTT
83
4498
|
|
1208673
N/A
N/A
1074
1089
ATGCACCAGGTGATGG
48
4499
|
|
1208686
N/A
N/A
1140
1155
AATGGCAGGTACCCTT
64
4500
|
|
1208699
N/A
N/A
1205
1220
AGGTAATTTAGTGTCT
96
4501
|
|
1208712
N/A
N/A
1550
1565
GTCCATCTGGTTCACA
69
4502
|
|
1208725
N/A
N/A
1581
1596
AACCCCACCAAGTCAG
55
4503
|
|
1208749
N/A
N/A
1629
1644
AGTCCAGGGTTGGAAT
45
4504
|
|
1208777
N/A
N/A
1700
1715
GGCACCAAGTTTGGGT
72
4505
|
|
1208803
N/A
N/A
1766
1781
AAGATGCAGATGGGAT
99
4506
|
|
1208828
N/A
N/A
2083
2098
ATGGACCTTTAAGATG
58
4507
|
|
1208855
N/A
N/A
2118
2133
CCAGAACTAGGACAGT
70
4508
|
|
1208882
N/A
N/A
2135
2150
ACTTGCCAAGGACCAA
40
4509
|
|
1208906
N/A
N/A
2173
2188
AATAAGCTTTTGCCAC
69
4510
|
|
1208932
N/A
N/A
2208
2223
AATGCTGAGCAGTGTC
96
4511
|
|
1208959
N/A
N/A
2278
2293
GCGAATAATGAGAATT
50
4512
|
|
1208985
N/A
N/A
2329
2344
ATACTGTTTGCTAGTT
81
4513
|
|
1209012
N/A
N/A
2379
2394
TATGCTGTTGGTTGAT
54
4514
|
|
1209036
N/A
N/A
2453
2468
TTAGCATTGATGAATC
50
4515
|
|
1209061
N/A
N/A
2482
2497
ATTCAGGTGTGTGCTA
47
4516
|
|
1209088
N/A
N/A
2528
2543
AGCACAAGAAGGCTAC
57
4517
|
|
1209112
N/A
N/A
2587
2602
GGGATTTGGCTTTTGG
76
4518
|
|
1209140
N/A
N/A
2699
2714
TTAAGGTTCAACAAGG
64
4519
|
|
1209169
N/A
N/A
2723
2738
AGTGAAAAATTTGCAC
70
4520
|
|
1209195
N/A
N/A
2779
2794
ACTCCCAAGTCAATAC
88
4521
|
|
1209216
N/A
N/A
2811
2826
AGAATGTGCCTACTTG
37
4522
|
|
1209243
N/A
N/A
2853
2868
TAATAGTACAGTTGAT
46
4523
|
|
1209266
N/A
N/A
2909
2924
CACTGTTAACCACTCT
84
4524
|
|
1209292
N/A
N/A
2990
3005
TGGGTAAGTCTTCAGT
62
4525
|
|
1209317
N/A
N/A
3044
3059
AGTCAGATAGCTGGAG
94
4526
|
|
1209341
N/A
N/A
3416
3431
CAGACCCTCAAAAAGG
64
4527
|
|
1209354
N/A
N/A
3467
3482
CTGTACTCAACTGCTC
47
4528
|
|
1209367
N/A
N/A
3497
3512
CAGGGACAACACACTC
61
4529
|
|
1209380
N/A
N/A
3648
3663
GGTACATGTCTCCCAG
42
4530
|
|
1209393
N/A
N/A
3682
3697
CCAAGCAGAGTTCCTT
74
4531
|
|
1209406
N/A
N/A
3758
3773
ACCTGTAGAAAGAGAC
41
4532
|
|
1209419
N/A
N/A
3893
3908
GATACCAGGAGAGTAA
70
4533
|
|
1209432
N/A
N/A
3960
3975
CTTCCAAAGGGTATTG
79
4534
|
|
1209445
N/A
N/A
4022
4037
TCCAAGCTTGGTTTAC
99
4535
|
|
1209458
N/A
N/A
4055
4070
CCTACTTGCCTTGTGA
67
4536
|
|
1209471
N/A
N/A
4130
4145
GTCTGAGAGATGGACA
54
4537
|
|
1209484
N/A
N/A
4305
4320
TATTCTGTAGGCCCAG
38
4538
|
|
1209497
N/A
N/A
4342
4357
TCTAGGACCATGCCAA
69
4539
|
|
1209510
N/A
N/A
4533
4548
GGCCCGTTCCCAACCA
45
4540
|
|
1209523
N/A
N/A
4641
4656
GAGGGAACGCAGTGAG
120
4541
|
|
1209536
N/A
N/A
4834
4849
TTTCCGCACTCTCCCT
78
4542
|
|
1209549
N/A
N/A
5382
5397
CCCCCCACTTCCTAAC
51
4543
|
|
1209562
N/A
N/A
5442
5457
AGCCCGCGGCCGGCTG
82
4544
|
|
1209575
N/A
N/A
5761
5776
CGCTGGGAGACGGAGG
31
4545
|
|
1209588
N/A
N/A
5913
5928
TCAGACCTGGCCACAA
45
4546
|
|
1209601
N/A
N/A
6326
6341
CCTAGCAGTTGTGCCT
70
4547
|
|
1209614
N/A
N/A
6404
6419
GGCACCCGGAACGATA
53
4548
|
|
1209627
N/A
N/A
6461
6476
CCTGGGATTCACCTAC
51
4549
|
|
1209640
N/A
N/A
6494
6509
TCCCAGCAGTGAATCC
49
4550
|
|
1209653
N/A
N/A
6575
6590
CTAGTTGGCCTTTGCA
52
4551
|
|
1209666
N/A
N/A
6638
6653
CACCACCCATCCAGAG
47
4552
|
|
1209679
N/A
N/A
6758
6773
CTCTCGCAGCAAGCCC
52
4553
|
|
1209692
N/A
N/A
6806
6821
ATGGACAAAGCTGCTC
29
4554
|
|
1209705
N/A
N/A
6872
6887
CCTCCGCCCCTGCGAA
60
4555
|
|
1209718
N/A
N/A
7070
7085
ACCTGAGATTTGTGCC
65†
4556
|
|
TABLE 71
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with
|
uniform PS internucleoside linkages (Huh7,
|
electroporation, 2000 nM modified oligonucleotide)
|
SEQ ID
SEQ ID
SEQ ID
SEQ ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
Sequence
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
35
2426
|
|
1129531
2009
2024
7397
7412
GAGTTCCTGCGCCATC
14
3837
|
|
1129948
N/A
N/A
4840
4855
ATCTGCTTTCCGCACT
31
3630
|
|
1194292
N/A
N/A
4843
4858
CTCATCTGCTTTCCGC
65
4557
|
|
1208277
N/A
N/A
121
136
TGGGACAATCCTGGTT
92
4558
|
|
1208312
N/A
N/A
183
198
CCCACAGGTCATGAGC
63
4559
|
|
1208347
N/A
N/A
233
248
GCCTTCCGGGCTGGCC
57
4560
|
|
1208378
N/A
N/A
310
325
TTAAGGCCCATCTCCC
111
4561
|
|
1208408
N/A
N/A
358
373
GATTTCTTCCCAAGAC
49
4562
|
|
1208439
N/A
N/A
430
445
TGACTATAAGTCATAG
43
4563
|
|
1208473
N/A
N/A
472
487
TCCCAAGGTGGAATCT
75
4564
|
|
1208502
N/A
N/A
572
587
TGCACCATACACATCC
59
4565
|
|
1208529
N/A
N/A
621
636
CCTAGCACCAGGTAGG
40
4566
|
|
1208562
N/A
N/A
651
666
ACGATCACTCTAGTGC
78
4567
|
|
1208583
N/A
N/A
717
732
GCTCCGAGCCAGGCTC
81
4568
|
|
1208596
N/A
N/A
836
851
TGAATGATGCCCATGA
41
4569
|
|
1208609
N/A
N/A
872
887
TCCTTGCCCATTGAAG
72
4570
|
|
1208622
N/A
N/A
909
924
AGAGGCATTCAGATGC
109
4571
|
|
1208635
N/A
N/A
952
967
TCGCCCAGAGTCACCC
55
4572
|
|
1208648
N/A
N/A
1008
1023
TGTTATCAACCTGGTT
42
4573
|
|
1208661
N/A
N/A
1036
1051
CCCTTCTCAACAACCC
76
4574
|
|
1208674
N/A
N/A
1076
1091
CCATGCACCAGGTGAT
52
4575
|
|
1208687
N/A
N/A
1141
1156
AAATGGCAGGTACCCT
103
4576
|
|
1208700
N/A
N/A
1209
1224
ATGTAGGTAATTTAGT
76
4577
|
|
1208713
N/A
N/A
1552
1567
AGGTCCATCTGGTTCA
84
4578
|
|
1208726
N/A
N/A
1582
1597
CAACCCCACCAAGTCA
45
4579
|
|
1208752
N/A
N/A
1630
1645
GAGTCCAGGGTTGGAA
26
4580
|
|
1208779
N/A
N/A
1703
1718
TAGGGCACCAAGTTTG
27
4581
|
|
1208805
N/A
N/A
1913
1928
GAAGTTGTGTGCCTGT
64
4582
|
|
1208829
N/A
N/A
2085
2100
AGATGGACCTTTAAGA
34
4583
|
|
1208856
N/A
N/A
2119
2134
ACCAGAACTAGGACAG
57
4584
|
|
1208884
N/A
N/A
2136
2151
CACTTGCCAAGGACCA
47
4585
|
|
1208909
N/A
N/A
2174
2189
CAATAAGCTTTTGCCA
78
4586
|
|
1208933
N/A
N/A
2214
2229
CCATGAAATGCTGAGC
29
4587
|
|
1208960
N/A
N/A
2286
2301
TAACCATCGCGAATAA
52
4588
|
|
1208988
N/A
N/A
2331
2346
TAATACTGTTTGCTAG
48
4589
|
|
1209013
N/A
N/A
2384
2399
GATTATATGCTGTTGG
82
4590
|
|
1209039
N/A
N/A
2454
2469
CTTAGCATTGATGAAT
51
4591
|
|
1209064
N/A
N/A
2484
2499
TGATTCAGGTGTGTGC
75
4592
|
|
1209089
N/A
N/A
2537
2552
TAGTTCCTAAGCACAA
32
4593
|
|
1209115
N/A
N/A
2614
2629
CGTTTTTCACACTTTG
40
4594
|
|
1209142
N/A
N/A
2701
2716
AGTTAAGGTTCAACAA
55
4595
|
|
1209170
N/A
N/A
2725
2740
GCAGTGAAAAATTTGC
62
4596
|
|
1209196
N/A
N/A
2780
2795
AACTCCCAAGTCAATA
30
4597
|
|
1209219
N/A
N/A
2812
2827
GAGAATGTGCCTACTT
40
4598
|
|
1209244
N/A
N/A
2864
2879
AACGGCAGTAATAATA
69
4599
|
|
1209269
N/A
N/A
2914
2929
GAAGTCACTGTTAACC
65
4600
|
|
1209293
N/A
N/A
2993
3008
CCTTGGGTAAGTCTTC
86
4601
|
|
1209320
N/A
N/A
3050
3065
TTCAGGAGTCAGATAG
87
4602
|
|
1209342
N/A
N/A
3419
3434
GGACAGACCCTCAAAA
54
4603
|
|
1209355
N/A
N/A
3468
3483
CCTGTACTCAACTGCT
36
4604
|
|
1209368
N/A
N/A
3498
3513
GCAGGGACAACACACT
53
4605
|
|
1209381
N/A
N/A
3650
3665
AGGGTACATGTCTCCC
93
4606
|
|
1209394
N/A
N/A
3684
3699
CTCCAAGCAGAGTTCC
56
4607
|
|
1209407
N/A
N/A
3760
3775
ACACCTGTAGAAAGAG
58
4608
|
|
1209420
N/A
N/A
3894
3909
TGATACCAGGAGAGTA
61
4609
|
|
1209433
N/A
N/A
3961
3976
ACTTCCAAAGGGTATT
26
4610
|
|
1209446
N/A
N/A
4023
4038
TTCCAAGCTTGGTTTA
48
4611
|
|
1209459
N/A
N/A
4056
4071
ACCTACTTGCCTTGTG
31
4612
|
|
1209472
N/A
N/A
4131
4146
GGTCTGAGAGATGGAC
41
4613
|
|
1209485
N/A
N/A
4307
4322
CCTATTCTGTAGGCCC
71
4614
|
|
1209498
N/A
N/A
4346
4361
AGAGTCTAGGACCATG
50
4615
|
|
1209511
N/A
N/A
4577
4592
ACCACCCGGCCTCCTG
71
4616
|
|
1209524
N/A
N/A
4642
4657
GGAGGGAACGCAGTGA
113
4617
|
|
1209537
N/A
N/A
4909
4924
TTGCCTTGGTGTCTGA
40
4618
|
|
1209550
N/A
N/A
5383
5398
CCCCCCCACTTCCTAA
68
4619
|
|
1209563
N/A
N/A
5444
5459
AGAGCCCGCGGCCGGC
79
4620
|
|
1209576
N/A
N/A
5767
5782
AAGCTGCGCTGGGAGA
43
4621
|
|
1209589
N/A
N/A
5918
5933
CGCTCTCAGACCTGGC
79
4622
|
|
1209602
N/A
N/A
6336
6351
TACCCCTGCCCCTAGC
51
4623
|
|
1209615
N/A
N/A
6411
6426
TTCTGTAGGCACCCGG
49
4624
|
|
1209628
N/A
N/A
6465
6480
TCTACCTGGGATTCAC
84
4625
|
|
1209641
N/A
N/A
6495
6510
ATCCCAGCAGTGAATC
111
4626
|
|
1209654
N/A
N/A
6601
6616
CTAGCCCGGAGCGCGG
66
4627
|
|
1209667
N/A
N/A
6660
6675
GCGGAAACAGAAACCC
49
4628
|
|
1209680
N/A
N/A
6764
6779
CTTCCTCTCTCGCAGC
60
4629
|
|
1209693
N/A
N/A
6807
6822
GATGGACAAAGCTGCT
87
4630
|
|
1209706
N/A
N/A
6875
6890
ATTCCTCCGCCCCTGC
76
4631
|
|
1209719
N/A
N/A
7071
7086
GACCTGAGATTTGTGC
98†
4632
|
|
TABLE 72
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with
|
uniform PS internucleoside linkages (Huh7,
|
electroporation, 2000 nM modified oligonucleotide)
|
SEQ ID
SEQ ID
SEQ ID
SEQ ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
Sequence
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
20
2426
|
|
1129531
2009
2024
7397
7412
GAGTTCCTGCGCCATC
8
3837
|
|
1129948
N/A
N/A
4840
4855
ATCTGCTTTCCGCACT
21
3630
|
|
1129949
N/A
N/A
4844
4859
TCTCATCTGCTTTCCG
27
3706
|
|
1208281
N/A
N/A
122
137
CTGGGACAATCCTGGT
104
4633
|
|
1208314
N/A
N/A
184
199
ACCCACAGGTCATGAG
28
4634
|
|
1208349
N/A
N/A
234
249
GGCCTTCCGGGCTGGC
59
4635
|
|
1208379
N/A
N/A
311
326
CTTAAGGCCCATCTCC
37
4636
|
|
1208412
N/A
N/A
398
413
TGCTAGTCTGCAGCTT
35
4637
|
|
1208443
N/A
N/A
431
446
CTGACTATAAGTCATA
47
4638
|
|
1208476
N/A
N/A
517
532
CCGACTGTGTGCTCTT
13
4639
|
|
1208504
N/A
N/A
573
588
CTGCACCATACACATC
27
4640
|
|
1208530
N/A
N/A
622
637
ACCTAGCACCAGGTAG
85
4641
|
|
1208564
N/A
N/A
656
671
AGCTCACGATCACTCT
86
4642
|
|
1208584
N/A
N/A
718
733
TGCTCCGAGCCAGGCT
45
4643
|
|
1208597
N/A
N/A
837
852
ATGAATGATGCCCATG
35
4644
|
|
1208610
N/A
N/A
878
893
GATCCTTCCTTGCCCA
27
4645
|
|
1208623
N/A
N/A
913
928
CTTCAGAGGCATTCAG
65
4646
|
|
1208636
N/A
N/A
954
969
CCTCGCCCAGAGTCAC
109
4647
|
|
1208649
N/A
N/A
1011
1026
TACTGTTATCAACCTG
40
4648
|
|
1208662
N/A
N/A
1046
1061
ACATGGGTTTCCCTTC
52
4649
|
|
1208675
N/A
N/A
1078
1093
TTCCATGCACCAGGTG
34
4650
|
|
1208688
N/A
N/A
1142
1157
GAAATGGCAGGTACCC
47
4651
|
|
1208701
N/A
N/A
1210
1225
CATGTAGGTAATTTAG
21
4652
|
|
1208714
N/A
N/A
1554
1569
TAAGGTCCATCTGGTT
92
4653
|
|
1208729
N/A
N/A
1584
1599
CTCAACCCCACCAAGT
79
4654
|
|
1208753
N/A
N/A
1667
1682
AAACTTTTGGGTGTGG
35
4655
|
|
1208782
N/A
N/A
1705
1720
GATAGGGCACCAAGTT
85
4656
|
|
1208806
N/A
N/A
1915
1930
ATGAAGTTGTGTGCCT
48
4657
|
|
1208832
N/A
N/A
2086
2101
GAGATGGACCTTTAAG
38
4658
|
|
1208859
N/A
N/A
2122
2137
CAAACCAGAACTAGGA
58
4659
|
|
1208887
N/A
N/A
2142
2157
AAAGTGCACTTGCCAA
29
4660
|
|
1208910
N/A
N/A
2175
2190
TCAATAAGCTTTTGCC
31
4661
|
|
1208936
N/A
N/A
2215
2230
GCCATGAAATGCTGAG
24
4662
|
|
1208963
N/A
N/A
2294
2309
ATAGAACATAACCATC
35
4663
|
|
1208989
N/A
N/A
2333
2348
CCTAATACTGTTTGCT
21
4664
|
|
1209016
N/A
N/A
2385
2400
AGATTATATGCTGTTG
23
4665
|
|
1209040
N/A
N/A
2455
2470
GCTTAGCATTGATGAA
51
4666
|
|
1209065
N/A
N/A
2486
2501
CTTGATTCAGGTGTGT
31
4667
|
|
1209092
N/A
N/A
2538
2553
GTAGTTCCTAAGCACA
48
4668
|
|
1209118
N/A
N/A
2630
2645
AGTCTACTTAGTGCAA
39
4669
|
|
1209144
N/A
N/A
2702
2717
CAGTTAAGGTTCAACA
26
4670
|
|
1209173
N/A
N/A
2744
2759
ATTTGTGGGCATGCAC
57
4671
|
|
1209199
N/A
N/A
2781
2796
TAACTCCCAAGTCAAT
49
4672
|
|
1209220
N/A
N/A
2813
2828
TGAGAATGTGCCTACT
24
4673
|
|
1209246
N/A
N/A
2865
2880
AAACGGCAGTAATAAT
59
4674
|
|
1209270
N/A
N/A
2917
2932
GTTGAAGTCACTGTTA
23
4675
|
|
1209296
N/A
N/A
3013
3028
ACAACCTGCTAGCTGT
73
4676
|
|
1209321
N/A
N/A
3051
3066
GTTCAGGAGTCAGATA
57
4677
|
|
1209343
N/A
N/A
3421
3436
AAGGACAGACCCTCAA
60
4678
|
|
1209356
N/A
N/A
3470
3485
CCCCTGTACTCAACTG
64
4679
|
|
1209369
N/A
N/A
3499
3514
TGCAGGGACAACACAC
82
4680
|
|
1209382
N/A
N/A
3651
3666
CAGGGTACATGTCTCC
39
4681
|
|
1209395
N/A
N/A
3686
3701
CTCTCCAAGCAGAGTT
74
4682
|
|
1209408
N/A
N/A
3762
3777
GCACACCTGTAGAAAG
54
4683
|
|
1209421
N/A
N/A
3896
3911
GGTGATACCAGGAGAG
56
4684
|
|
1209434
N/A
N/A
3962
3977
GACTTCCAAAGGGTAT
57
4685
|
|
1209447
N/A
N/A
4025
4040
GTTTCCAAGCTTGGTT
55
4686
|
|
1209460
N/A
N/A
4057
4072
AACCTACTTGCCTTGT
28
4687
|
|
1209473
N/A
N/A
4242
4257
CACCTTTCTGGCAGTG
56
4688
|
|
1209486
N/A
N/A
4317
4332
TATCCAGCAACCTATT
45
4689
|
|
1209499
N/A
N/A
4351
4366
TCAGGAGAGTCTAGGA
42
4690
|
|
1209512
N/A
N/A
4579
4594
ACACCACCCGGCCTCC
30
4691
|
|
1209525
N/A
N/A
4657
4672
GTGCGGCAGGCTTGGG
27
4692
|
|
1209538
N/A
N/A
5077
5092
CCCACGCGGCGCACCG
57
4693
|
|
1209551
N/A
N/A
5404
5419
GCCCTCTCGGCTCCTC
33
4694
|
|
1209564
N/A
N/A
5445
5460
GAGAGCCCGCGGCCGG
60
4695
|
|
1209577
N/A
N/A
5771
5786
GTGGAAGCTGCGCTGG
30
4696
|
|
1209590
N/A
N/A
5919
5934
GCGCTCTCAGACCTGG
111
4697
|
|
1209603
N/A
N/A
6352
6367
CAAAGGTCTCCTCCCC
54
4698
|
|
1209616
N/A
N/A
6414
6429
CCATTCTGTAGGCACC
25
4699
|
|
1209629
N/A
N/A
6468
6483
GATTCTACCTGGGATT
70
4700
|
|
1209642
N/A
N/A
6496
6511
GATCCCAGCAGTGAAT
74
4701
|
|
1209655
N/A
N/A
6607
6622
CCCACACTAGCCCGGA
80
4702
|
|
1209668
N/A
N/A
6661
6676
AGCGGAAACAGAAACC
83
4703
|
|
1209681
N/A
N/A
6766
6781
CCCTTCCTCTCTCGCA
67
4704
|
|
1209694
N/A
N/A
6808
6823
CGATGGACAAAGCTGC
40
4705
|
|
1209707
N/A
N/A
7014
7029
ACCTGGCACGCATCGG
10†
4706
|
|
1209720
N/A
N/A
7073
7088
TGGACCTGAGATTTGT
44†
4707
|
|
TABLE 73
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with
|
uniform PS internucleoside linkages (Huh7,
|
electroporation, 2000 nM modified oligonucleotide)
|
SEQ ID
SEQ ID
SEQ ID
SEQ ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
Sequence
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
42
2426
|
|
1129531
2009
2024
7397
7412
GAGTTCCTGCGCCATC
12
3837
|
|
1129948
N/A
N/A
4840
4855
ATCTGCTTTCCGCACT
25
3630
|
|
1194293
N/A
N/A
4845
4860
CTCTCATCTGCTTTCC
46
4708
|
|
1208282
N/A
N/A
124
139
TCCTGGGACAATCCTG
94
4709
|
|
1208317
N/A
N/A
185
200
GACCCACAGGTCATGA
41
4710
|
|
1208351
N/A
N/A
265
280
ACCCAGTCTGGTTGTC
54
4711
|
|
1208382
N/A
N/A
312
327
TCTTAAGGCCCATCTC
78
4712
|
|
1208413
N/A
N/A
399
414
TTGCTAGTCTGCAGCT
22
4713
|
|
1208444
N/A
N/A
432
447
ACTGACTATAAGTCAT
79
4714
|
|
1208478
N/A
N/A
518
533
ACCGACTGTGTGCTCT
26
4715
|
|
1208506
N/A
N/A
576
591
ACACTGCACCATACAC
18
4716
|
|
1208534
N/A
N/A
625
640
AAGACCTAGCACCAGG
25
4717
|
|
1208566
N/A
N/A
657
672
CAGCTCACGATCACTC
40
4718
|
|
1208585
N/A
N/A
721
736
ACCTGCTCCGAGCCAG
79
4719
|
|
1208598
N/A
N/A
838
853
CATGAATGATGCCCAT
36
4720
|
|
1208611
N/A
N/A
881
896
CTTGATCCTTCCTTGC
39
4721
|
|
1208624
N/A
N/A
914
929
GCTTCAGAGGCATTCA
45
4722
|
|
1208637
N/A
N/A
961
976
TCAAATCCCTCGCCCA
63
4723
|
|
1208650
N/A
N/A
1012
1027
TTACTGTTATCAACCT
34
4724
|
|
1208663
N/A
N/A
1048
1063
TCACATGGGTTTCCCT
30
4725
|
|
1208676
N/A
N/A
1080
1095
ATTTCCATGCACCAGG
105
4726
|
|
1208689
N/A
N/A
1143
1158
GGAAATGGCAGGTACC
26
4727
|
|
1208702
N/A
N/A
1211
1226
GCATGTAGGTAATTTA
44
4728
|
|
1208715
N/A
N/A
1563
1578
CCACCAGGGTAAGGTC
30
4729
|
|
1208730
N/A
N/A
1585
1600
ACTCAACCCCACCAAG
143
4730
|
|
1208756
N/A
N/A
1669
1684
AGAAACTTTTGGGTGT
55
4731
|
|
1208784
N/A
N/A
1706
1721
GGATAGGGCACCAAGT
110
4732
|
|
1208808
N/A
N/A
1918
1933
AGCATGAAGTTGTGTG
68
4733
|
|
1208833
N/A
N/A
2087
2102
TGAGATGGACCTTTAA
63
4734
|
|
1208860
N/A
N/A
2123
2138
CCAAACCAGAACTAGG
26
4735
|
|
1208888
N/A
N/A
2144
2159
GCAAAGTGCACTTGCC
79
4736
|
|
1208913
N/A
N/A
2177
2192
GCTCAATAAGCTTTTG
29
4737
|
|
1208937
N/A
N/A
2216
2231
TGCCATGAAATGCTGA
32
4738
|
|
1208964
N/A
N/A
2316
2331
GTTCAATGTTCACTGT
31
4739
|
|
1208992
N/A
N/A
2334
2349
ACCTAATACTGTTTGC
59
4740
|
|
1209017
N/A
N/A
2386
2401
CAGATTATATGCTGTT
18
4741
|
|
1209043
N/A
N/A
2456
2471
AGCTTAGCATTGATGA
38
4742
|
|
1209068
N/A
N/A
2488
2503
CACTTGATTCAGGTGT
66
4743
|
|
1209093
N/A
N/A
2539
2554
TGTAGTTCCTAAGCAC
35
4744
|
|
1209119
N/A
N/A
2633
2648
CTCAGTCTACTTAGTG
16
4745
|
|
1209147
N/A
N/A
2704
2719
CCCAGTTAAGGTTCAA
40
4746
|
|
1209174
N/A
N/A
2745
2760
CATTTGTGGGCATGCA
41
4747
|
|
1209200
N/A
N/A
2782
2797
GTAACTCCCAAGTCAA
64
4748
|
|
1209247
N/A
N/A
2868
2883
GTAAAACGGCAGTAAT
56
4749
|
|
1209273
N/A
N/A
2924
2939
GTTGAATGTTGAAGTC
53
4750
|
|
1209297
N/A
N/A
3014
3029
CACAACCTGCTAGCTG
36
4751
|
|
1209324
N/A
N/A
3059
3074
AAACTTGGGTTCAGGA
43
4752
|
|
1209344
N/A
N/A
3422
3437
AAAGGACAGACCCTCA
55
4753
|
|
1209357
N/A
N/A
3471
3486
TCCCCTGTACTCAACT
123
4754
|
|
1209370
N/A
N/A
3501
3516
ACTGCAGGGACAACAC
26
4755
|
|
1209383
N/A
N/A
3654
3669
AGGCAGGGTACATGTC
55
4756
|
|
1209396
N/A
N/A
3700
3715
TATCACAGTCCCCTCT
45
4757
|
|
1209409
N/A
N/A
3836
3851
GTGTAGCACCTTTCAC
58
4758
|
|
1209422
N/A
N/A
3898
3913
CTGGTGATACCAGGAG
17
4759
|
|
1209435
N/A
N/A
3975
3990
AACTCTCCCTCTGGAC
41
4760
|
|
1209448
N/A
N/A
4026
4041
AGTTTCCAAGCTTGGT
26
4761
|
|
1209461
N/A
N/A
4063
4078
TTCTTGAACCTACTTG
96
4762
|
|
1209474
N/A
N/A
4243
4258
TCACCTTTCTGGCAGT
33
4763
|
|
1209487
N/A
N/A
4319
4334
AGTATCCAGCAACCTA
42
4764
|
|
1209500
N/A
N/A
4352
4367
CTCAGGAGAGTCTAGG
79
4765
|
|
1209513
N/A
N/A
4605
4620
CCCCCCAGAGAGCTCT
40
4766
|
|
1209526
N/A
N/A
4660
4675
TTGGTGCGGCAGGCTT
26
4767
|
|
1209539
N/A
N/A
5079
5094
GCCCCACGCGGCGCAC
46
4768
|
|
1209552
N/A
N/A
5405
5420
CGCCCTCTCGGCTCCT
52
4769
|
|
1209565
N/A
N/A
5454
5469
CTGAGGACGGAGAGCC
19
4770
|
|
1209578
N/A
N/A
5802
5817
GGAGAAGGTAGGGCAC
101
4771
|
|
1209591
N/A
N/A
6090
6105
CCCACGCACCCAGGTC
57
4772
|
|
1209604
N/A
N/A
6353
6368
TCAAAGGTCTCCTCCC
100
4773
|
|
1209617
N/A
N/A
6416
6431
ACCCATTCTGTAGGCA
73
4774
|
|
1209630
N/A
N/A
6469
6484
GGATTCTACCTGGGAT
33
4775
|
|
1209643
N/A
N/A
6516
6531
GTATCCCCAGGAGATT
59
4776
|
|
1209656
N/A
N/A
6609
6624
CTCCCACACTAGCCCG
91
4777
|
|
1209669
N/A
N/A
6662
6677
GAGCGGAAACAGAAAC
75
4778
|
|
1209682
N/A
N/A
6777
6792
ACCCACTCATGCCCTT
76
4779
|
|
1209695
N/A
N/A
6820
6835
TTGCCGCCCGGACGAT
31
4780
|
|
1209708
N/A
N/A
7023
7038
TAAGAGCTCACCTGGC
57†
4781
|
|
1209721
N/A
N/A
7079
7094
GCGCTGTGGACCTGAG
37†
4782
|
|
TABLE 74
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with
|
uniform PS internucleoside linkages (Huh7,
|
electroporation, 2000 nM modified oligonucleotide)
|
SEQ ID
SEQ ID
SEQ ID
SEQ ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
Sequence
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
29
2426
|
|
1129531
2009
2024
7397
7412
GAGTTCCTGCGCCATC
18
3837
|
|
1129948
N/A
N/A
4840
4855
ATCTGCTTTCCGCACT
54
3630
|
|
1194294
N/A
N/A
4846
4861
CCTCTCATCTGCTTTC
27
4783
|
|
1208286
N/A
N/A
125
140
ATCCTGGGACAATCCT
50
4784
|
|
1208319
N/A
N/A
188
203
CTGGACCCACAGGTCA
42
4785
|
|
1208354
N/A
N/A
266
281
CACCCAGTCTGGTTGT
49
4786
|
|
1208383
N/A
N/A
313
328
TTCTTAAGGCCCATCT
79
4787
|
|
1208417
N/A
N/A
400
415
GTTGCTAGTCTGCAGC
58
4788
|
|
1208448
N/A
N/A
433
448
AACTGACTATAAGTCA
25
4789
|
|
1208480
N/A
N/A
519
534
TACCGACTGTGTGCTC
49
4790
|
|
1208508
N/A
N/A
578
593
ACACACTGCACCATAC
22
4791
|
|
1208535
N/A
N/A
627
642
ATAAGACCTAGCACCA
40
4792
|
|
1208568
N/A
N/A
658
673
ACAGCTCACGATCACT
30
4793
|
|
1208586
N/A
N/A
724
739
AGCACCTGCTCCGAGC
81
4794
|
|
1208599
N/A
N/A
839
854
GCATGAATGATGCCCA
55
4795
|
|
1208612
N/A
N/A
885
900
AGCTCTTGATCCTTCC
18
4796
|
|
1208625
N/A
N/A
915
930
GGCTTCAGAGGCATTC
57
4797
|
|
1208638
N/A
N/A
962
977
CTCAAATCCCTCGCCC
27
4798
|
|
1208651
N/A
N/A
1013
1028
TTTACTGTTATCAACC
22
4799
|
|
1208664
N/A
N/A
1049
1064
CTCACATGGGTTTCCC
52
4800
|
|
1208677
N/A
N/A
1122
1137
TCTAAAAGGTAGTTTC
27
4801
|
|
1208690
N/A
N/A
1144
1159
GGGAAATGGCAGGTAC
30
4802
|
|
1208703
N/A
N/A
1212
1227
GGCATGTAGGTAATTT
44
4803
|
|
1208716
N/A
N/A
1564
1579
CCCACCAGGGTAAGGT
147
4804
|
|
1208731
N/A
N/A
1589
1604
AGAGACTCAACCCCAC
50
4805
|
|
1208757
N/A
N/A
1670
1685
GAGAAACTTTTGGGTG
19
4806
|
|
1208786
N/A
N/A
1714
1729
TTAGGCAAGGATAGGG
79
4807
|
|
1208809
N/A
N/A
1919
1934
GAGCATGAAGTTGTGT
73
4808
|
|
1208836
N/A
N/A
2088
2103
CTGAGATGGACCTTTA
26
4809
|
|
1208863
N/A
N/A
2124
2139
ACCAAACCAGAACTAG
24
4810
|
|
1208891
N/A
N/A
2147
2162
AAGGCAAAGTGCACTT
42
4811
|
|
1208914
N/A
N/A
2179
2194
CTGCTCAATAAGCTTT
96
4812
|
|
1208940
N/A
N/A
2219
2234
TAATGCCATGAAATGC
58
4813
|
|
1208967
N/A
N/A
2318
2333
TAGTTCAATGTTCACT
27
4814
|
|
1208993
N/A
N/A
2335
2350
AACCTAATACTGTTTG
30
4815
|
|
1209020
N/A
N/A
2387
2402
CCAGATTATATGCTGT
40
4816
|
|
1209044
N/A
N/A
2458
2473
TCAGCTTAGCATTGAT
52
4817
|
|
1209069
N/A
N/A
2491
2506
AGACACTTGATTCAGG
21
4818
|
|
1209096
N/A
N/A
2542
2557
CTGTGTAGTTCCTAAG
28
4819
|
|
1209122
N/A
N/A
2639
2654
GTCCTTCTCAGTCTAC
32
4820
|
|
1209148
N/A
N/A
2705
2720
TCCCAGTTAAGGTTCA
22
4821
|
|
1209176
N/A
N/A
2747
2762
GCCATTTGTGGGCATG
24
4822
|
|
1209203
N/A
N/A
2783
2798
TGTAACTCCCAAGTCA
80
4823
|
|
1209224
N/A
N/A
2815
2830
ATTGAGAATGTGCCTA
39
4824
|
|
1209249
N/A
N/A
2869
2884
TGTAAAACGGCAGTAA
27
4825
|
|
1209274
N/A
N/A
2954
2969
TTGCCACAAAGTAGGC
45
4826
|
|
1209299
N/A
N/A
3015
3030
CCACAACCTGCTAGCT
57
4827
|
|
1209325
N/A
N/A
3060
3075
AAAACTTGGGTTCAGG
60
4828
|
|
1209345
N/A
N/A
3423
3438
AAAAGGACAGACCCTC
72
4829
|
|
1209358
N/A
N/A
3472
3487
TTCCCCTGTACTCAAC
53
4830
|
|
1209371
N/A
N/A
3502
3517
AACTGCAGGGACAACA
140
4831
|
|
1209384
N/A
N/A
3655
3670
CAGGCAGGGTACATGT
27
4832
|
|
1209397
N/A
N/A
3701
3716
CTATCACAGTCCCCTC
39
4833
|
|
1209410
N/A
N/A
3872
3887
CGGGAGGAGAGAGCCC
37
4834
|
|
1209423
N/A
N/A
3918
3933
GAATCCCAGGTGTGTG
74
4835
|
|
1209436
N/A
N/A
3994
4009
TGGGACCACTCCTTCC
48
4836
|
|
1209449
N/A
N/A
4027
4042
AAGTTTCCAAGCTTGG
24
4837
|
|
1209462
N/A
N/A
4064
4079
CTTCTTGAACCTACTT
38
4838
|
|
1209475
N/A
N/A
4265
4280
CGCCCAGGTCCTCCAC
59
4839
|
|
1209488
N/A
N/A
4327
4342
AGTCTCCGAGTATCCA
112
4840
|
|
1209501
N/A
N/A
4353
4368
TCTCAGGAGAGTCTAG
56
4841
|
|
1209514
N/A
N/A
4606
4621
CCCCCCCAGAGAGCTC
20
4842
|
|
1209527
N/A
N/A
4765
4780
GACCCTCACTCACCCA
72
4843
|
|
1209540
N/A
N/A
5081
5096
CAGCCCCACGCGGCGC
76
4844
|
|
1209553
N/A
N/A
5406
5421
GCGCCCTCTCGGCTCC
107
4845
|
|
1209566
N/A
N/A
5455
5470
GCTGAGGACGGAGAGC
41
4846
|
|
1209579
N/A
N/A
5845
5860
CTCCCCGGGAGCTCCG
68
4847
|
|
1209592
N/A
N/A
6091
6106
CCCCACGCACCCAGGT
47
4848
|
|
1209605
N/A
N/A
6354
6369
ATCAAAGGTCTCCTCC
30
4849
|
|
1209618
N/A
N/A
6421
6436
GCGCCACCCATTCTGT
35
4850
|
|
1209631
N/A
N/A
6472
6487
CTGGGATTCTACCTGG
30
4851
|
|
1209644
N/A
N/A
6526
6541
GATTCTCCCTGTATCC
69
4852
|
|
1209657
N/A
N/A
6616
6631
AACCTGGCTCCCACAC
40
4853
|
|
1209670
N/A
N/A
6663
6678
GGAGCGGAAACAGAAA
105
4854
|
|
1209683
N/A
N/A
6778
6793
AACCCACTCATGCCCT
108
4855
|
|
1209696
N/A
N/A
6824
6839
ACGCTTGCCGCCCGGA
32
4856
|
|
1209709
N/A
N/A
7024
7039
CTAAGAGCTCACCTGG
82†
4857
|
|
1209722
N/A
N/A
7092
7107
AACACGCAGCTCAGCG
63†
4858
|
|
TABLE 75
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with
|
uniform PS internucleoside linkages (Huh7,
|
electroporation, 2000 nM modified oligonucleotide)
|
SEQ ID
SEQ ID
SEQ ID
SEQ ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
Sequence
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1128802
218
233
3569
3584
TACATTTGTGGTACAG
94
2426
|
|
1129531
2009
2024
7397
7412
GAGTTCCTGCGCCATC
12
3837
|
|
1129948
N/A
N/A
4840
4855
ATCTGCTTTCCGCACT
45
3630
|
|
1194295
N/A
N/A
4847
4862
CCCTCTCATCTGCTTT
46
4859
|
|
1208287
N/A
N/A
129
144
AACAATCCTGGGACAA
49
4860
|
|
1208322
N/A
N/A
194
209
AGTCACCTGGACCCAC
33
4861
|
|
1208356
N/A
N/A
267
282
CCACCCAGTCTGGTTG
76
4862
|
|
1208387
N/A
N/A
314
329
CTTCTTAAGGCCCATC
52
4863
|
|
1208418
N/A
N/A
402
417
CTGTTGCTAGTCTGCA
72
4864
|
|
1208449
N/A
N/A
438
453
CAGGGAACTGACTATA
42
4865
|
|
1208483
N/A
N/A
525
540
GCCACTTACCGACTGT
50
4866
|
|
1208509
N/A
N/A
585
600
AGACTGCACACACTGC
80
4867
|
|
1208539
N/A
N/A
628
643
CATAAGACCTAGCACC
54
4868
|
|
1208570
N/A
N/A
661
676
CACACAGCTCACGATC
61
4869
|
|
1208587
N/A
N/A
733
748
CATATCTCAAGCACCT
40
4870
|
|
1208600
N/A
N/A
840
855
GGCATGAATGATGCCC
56
4871
|
|
1208613
N/A
N/A
886
901
AAGCTCTTGATCCTTC
102
4872
|
|
1208626
N/A
N/A
925
940
GATAAAGCTGGGCTTC
71
4873
|
|
1208639
N/A
N/A
968
983
GGAGAACTCAAATCCC
65
4874
|
|
1208652
N/A
N/A
1014
1029
GTTTACTGTTATCAAC
79
4875
|
|
1208665
N/A
N/A
1052
1067
TACCTCACATGGGTTT
43
4876
|
|
1208678
N/A
N/A
1124
1139
CATCTAAAAGGTAGTT
64
4877
|
|
1208691
N/A
N/A
1164
1179
ATGGCAGAGCTTGAGG
50
4878
|
|
1208704
N/A
N/A
1213
1228
TGGCATGTAGGTAATT
54
4879
|
|
1208717
N/A
N/A
1565
1580
GCCCACCAGGGTAAGG
46
4880
|
|
1208734
N/A
N/A
1595
1610
ATGCTTAGAGACTCAA
61
4881
|
|
1208760
N/A
N/A
1671
1686
GGAGAAACTTTTGGGT
42
4882
|
|
1208788
N/A
N/A
1715
1730
ATTAGGCAAGGATAGG
41
4883
|
|
1208812
N/A
N/A
1920
1935
TGAGCATGAAGTTGTG
108
4884
|
|
1208839
N/A
N/A
2091
2106
GATCTGAGATGGACCT
63
4885
|
|
1208865
N/A
N/A
2125
2140
GACCAAACCAGAACTA
69
4886
|
|
1208892
N/A
N/A
2148
2163
CAAGGCAAAGTGCACT
65
4887
|
|
1208917
N/A
N/A
2181
2196
ACCTGCTCAATAAGCT
58
4888
|
|
1208943
N/A
N/A
2226
2241
CATGAGATAATGCCAT
80
4889
|
|
1208968
N/A
N/A
2319
2334
CTAGTTCAATGTTCAC
62
4890
|
|
1208996
N/A
N/A
2336
2351
GAACCTAATACTGTTT
83
4891
|
|
1209021
N/A
N/A
2388
2403
ACCAGATTATATGCTG
43
4892
|
|
1209047
N/A
N/A
2460
2475
CATCAGCTTAGCATTG
36
4893
|
|
1209072
N/A
N/A
2492
2507
TAGACACTTGATTCAG
33
4894
|
|
1209097
N/A
N/A
2544
2559
AGCTGTGTAGTTCCTA
82
4895
|
|
1209123
N/A
N/A
2646
2661
AATGAGTGTCCTTCTC
135
4896
|
|
1209151
N/A
N/A
2706
2721
TTCCCAGTTAAGGTTC
75
4897
|
|
1209177
N/A
N/A
2748
2763
GGCCATTTGTGGGCAT
63
4898
|
|
1209204
N/A
N/A
2784
2799
TTGTAACTCCCAAGTC
55
4899
|
|
1209227
N/A
N/A
2816
2831
CATTGAGAATGTGCCT
37
4900
|
|
1209250
N/A
N/A
2870
2885
CTGTAAAACGGCAGTA
45
4901
|
|
1209276
N/A
N/A
2956
2971
ACTTGCCACAAAGTAG
76
4902
|
|
1209300
N/A
N/A
3017
3032
CTCCACAACCTGCTAG
32
4903
|
|
1209328
N/A
N/A
3370
3385
AAAAGTTGGGTTCAGG
43
4904
|
|
1209346
N/A
N/A
3425
3440
GGAAAAGGACAGACCC
35
4905
|
|
1209359
N/A
N/A
3473
3488
CTTCCCCTGTACTCAA
72
4906
|
|
1209372
N/A
N/A
3503
3518
GAACTGCAGGGACAAC
62
4907
|
|
1209385
N/A
N/A
3656
3671
ACAGGCAGGGTACATG
43
4908
|
|
1209398
N/A
N/A
3702
3717
CCTATCACAGTCCCCT
51
4909
|
|
1209411
N/A
N/A
3878
3893
ATGAGGCGGGAGGAGA
37
4910
|
|
1209424
N/A
N/A
3919
3934
AGAATCCCAGGTGTGT
50
4911
|
|
1209437
N/A
N/A
3997
4012
AAATGGGACCACTCCT
114
4912
|
|
1209450
N/A
N/A
4028
4043
CAAGTTTCCAAGCTTG
90
4913
|
|
1209463
N/A
N/A
4065
4080
CCTTCTTGAACCTACT
65
4914
|
|
1209476
N/A
N/A
4266
4281
CCGCCCAGGTCCTCCA
60
4915
|
|
1209489
N/A
N/A
4329
4344
CAAGTCTCCGAGTATC
71
4916
|
|
1209502
N/A
N/A
4354
4369
GTCTCAGGAGAGTCTA
43
4917
|
|
1209515
N/A
N/A
4608
4623
GACCCCCCCAGAGAGC
107
4918
|
|
1209528
N/A
N/A
4773
4788
TTGCCCCAGACCCTCA
70
4919
|
|
1209541
N/A
N/A
5118
5133
CGCCGGGAGCCCGGAG
80
4920
|
|
1209554
N/A
N/A
5427
5442
GGCCGGAATCTAGCTC
41
4921
|
|
1209567
N/A
N/A
5714
5729
CGGGTACTCGCCGGTC
58
4922
|
|
1209580
N/A
N/A
5847
5862
TCCTCCCCGGGAGCTC
64
4923
|
|
1209593
N/A
N/A
6094
6109
CGCCCCCACGCACCCA
92
4924
|
|
1209606
N/A
N/A
6356
6371
TGATCAAAGGTCTCCT
31
4925
|
|
1209619
N/A
N/A
6435
6450
AACCCATCAGGTCAGC
43
4926
|
|
1209632
N/A
N/A
6473
6488
CCTGGGATTCTACCTG
29
4927
|
|
1209645
N/A
N/A
6528
6543
TCGATTCTCCCTGTAT
28
4928
|
|
1209658
N/A
N/A
6617
6632
GAACCTGGCTCCCACA
55
4929
|
|
1209671
N/A
N/A
6679
6694
CAGGATTTGAATGGGC
31
4930
|
|
1209684
N/A
N/A
6779
6794
AAACCCACTCATGCCC
39
4931
|
|
1209697
N/A
N/A
6837
6852
ACCCCATCTGACAACG
65
4932
|
|
1209710
N/A
N/A
7025
7040
GCTAAGAGCTCACCTG
86†
4933
|
|
1209723
N/A
N/A
7108
7123
ATCACCCTGGGTCGGA
68†
4934
|
|
TABLE 76
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with
|
uniform PS internucleoside linkages (Huh7,
|
electroporation, 3000 nM modified oligonucleotide)
|
SEQ ID
SEQ ID
SEQ ID
SEQ ID
|
NO: 3
NO: 3
NO: 4
NO: 4
|
Compound
Start
Stop
Start
Stop
Sequence
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1128718
3437
3452
13
28
CTGCCTATCCAGGAGT
69
4935
|
|
1128719
3439
3454
15
30
AGCTGCCTATCCAGGA
33
4936
|
|
1128720
3449
3464
25
40
CCGTTGGTCCAGCTGC
22
4937
|
|
1128721
3450
3465
26
41
TCCGTTGGTCCAGCTG
49
4938
|
|
1128726
3459
3474
35
50
TGGCATCCGTCCGTTG
60
4939
|
|
1128727
3460
3475
36
51
ATGGCATCCGTCCGTT
60
4940
|
|
1128728
3463
3478
39
54
CTCATGGCATCCGTCC
66
4941
|
|
1128729
3464
3479
40
55
CCTCATGGCATCCGTC
85
4942
|
|
1128730
3465
3480
41
56
CCCTCATGGCATCCGT
72
4943
|
|
1128731
3468
3483
44
59
GAGCCCTCATGGCATC
60
4944
|
|
TABLE 77
|
|
Reduction of FXII RNA by 3-10-3 cEt gapmers with
|
uniform PS internucleoside linkages (Huh7,
|
electroporation, 3000 nM modified oligonucleotide)
|
SEQ ID
SEQ ID
SEQ ID
SEQ ID
|
NO: 3
NO: 3
NO: 4
NO: 4
|
Compound
Start
Stop
Start
Stop
Sequence
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1128724
3455
3470
31
46
ATCCGTCCGTTGGTCC
42
4945
|
|
1128725
3457
3472
33
48
GCATCCGTCCGTTGGT
59
4946
|
|
Example 4: Effects of Modified Oligonucleotides with Mixed Sugar Modifications and Mixed Backbones on Human FXII RNA In Vitro, Single Dose
Modified oligonucleotides complementary to an FXII nucleic acid were synthesized and tested for their effect on FXII RNA levels in vitro. The modified oligonucleotides were tested in a series of experiments using similar culture conditions. The results for each separate experiment are presented in separate tables below.
The chemistry notation column in the tables below specifies the specific chemistry notation for modified oligonucleotides; wherein subscript ‘d’ represents a 2′-β-D-deoxyribosyl sugar moiety, subscript ‘e’ represents a 2′-MOE sugar moiety, subscript ‘y’ represents a 2-O-methyl sugar moiety, subscript ‘k’ represents a cEt modified sugar moiety, subscript ‘s’ represents a phosphorothioate internucleoside linkage, subscript ‘o’ represents a phosphodiester internucleoside linkage, and superscript ‘m’ before the cytosine residue (mC) represents a 5-methyl cytosine.
“Start site” indicates the 5′-most nucleoside of the target sequence to which the modified oligonucleotide is complementary. “Stop site” indicates the 3′-most nucleoside of the target sequence to which the modified oligonucleotide is complementary. As shown in the tables below, the modified oligonucleotides are 100% complementary to one or more of human FXII target sequences including the human FXII mRNA sequence designated herein as SEQ ID NO: 1 (ENSEMBL ID ENST00000253496.3 from ENSEMBL version 99: January 2020), the human FXII genomic sequence, designated herein as SEQ ID NO: 2 (ENSEMBL ID ENSG00000131187.9 from ENSEML version 99: January 2020, human reference assembly version GRCh38.p13 located on the reverse strand of chromosome 5 from positions 177,402,140 to 177,409,576), the human FXII genomic sequence, designated herein as SEQ ID No. 3 (the complement of GENBANK Accession No. NC_000005.10 truncated from truncated from nucleotides 177399001 to 177413000), and the human FXII mRNA sequence designated herein as SEQ ID No: 4 (GENBANK Accession No. NM_0005053), ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular target sequence with 100% complementarity.
Cultured Huh7 cells, at a density of 20,000 cells per well, were transfected using electroporation with either 2000 nM or 4000 nM modified oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and FXII RNA levels were measured by quantitative real-time RTPCR. Human primer probe set RTS40528 (described herein above) was used to measure RNA levels. FXII RNA levels were normalized to total RNA content, as measured by RIBOGREEN®. Reduction of FXII RNA is presented in the tables below as percent FXII RNA relative to the amount in untreated control cells (% UTC). The symbol “†” indicates that the modified oligonucleotide is complementary to the target transcript within the amplicon region of the primer probe set. In such instances, additional assays using alternative primer probes must be performed to accurately assess the potency and efficacy of such modified oligonucleotides.
TABLE 78
|
|
Reduction of FXII RNA (Huh7, electroporation,
|
4000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
Sequence with Chemistry
FXII
SEQ ID
|
Number
Site
Site
Site
Site
notation (5′ to 3′)
(% UTC)
NO
|
|
1129531
2009
2024
7397
7412
GksAksGksTdsTdsmCdsCdsTdsGdsmCds
44
3837
|
GdsmCdsmCdsAksTksmCk
|
|
1206480
1782
1797
7170
7185
AksTksGkSmCdsmCdsTdsTdsGdsmCdsAds
60
4947
|
GdsGdsGdsTksGksAk
|
|
1206486
1915
1930
7303
7318
TksmCksAksmCdsmCdsAdsAdsGdsGdsAds
59
4948
|
GdsGdsGdsAksAksAk
|
|
1206491
1934
1949
7322
7337
mCksmCksAksmCdsTdsmCdsTdsmCdsTds
64
4949
|
mCdsAdsmCdsTdsGksmCksGk
|
|
1206495
141
156
508
523
TksGksmCksTdsmCdsTdsTdsmCdsAdsGds
64
4950
|
mCdsTdsTdsTksGksTk
|
|
1206901
144
159
511
526
GksTesGksTdsGdsmCdsTdsmCdsTdsTds
81
4951
|
mCdsTdsGesmCesTksCk
|
|
1206909
152
167
N/A
N/A
GksAesAksmCdsGdsAdsmCdsTdsGdsTds
53
3647
|
GdsTdsGesmCesTksmCk
|
|
1206917
209
224
3560
3575
GksGesTksAdsmCdsAdsGdsmCdsTdsGds
42
4952
|
mCdsmCdsGesGesTksGk
|
|
1206925
217
232
3568
3583
AksmCesAksTdsTdsTdGdsTdGdsGdsTds
64
3114
|
AdsmCesAesGksmCk
|
|
1206933
1777
1792
7165
7180
TksTesGksmCdsAdsGdsGdsGdsTdsGds
62
3375
|
AdsGdsmCesmCesGksGk
|
|
1206941
1785
1800
7173
7188
AksTesGksAdsTdsGdsmCdmCdsTdsTds
57
3603
|
GdsmCdsAesGesGksGk
|
|
1206949
1793
1808
7181
7196
mCksmCesmCksAdsGdsmCdsTdsGdsAds
64
4953
|
TdsGdsAdsTesGesmCksmCk
|
|
1206957
1899
1914
7287
7302
GksAesTkGdsAdsGdsTdsmCdsmCdsmCds
47
2537
|
TdsGdsAesGesmCksAk
|
|
1206965
1914
1929
7302
7317
mCksAesmCksmCdsAdsAdsGdsGdsAdsGds
60
4954
|
GdsGdsAesAesAksGk
|
|
1206973
1922
1937
7310
7325
TksGesmCksGdsGdsAdsAdsTdsmCdsAds
62
3455
|
mCdsmCdsAesAesGksGk
|
|
1206981
1930
1945
7318
7333
TksmCesTksmCdsTdsmCdsAdsmCdsTds
50
4955
|
GdsmCdsGdsGesAesAksTk
|
|
1206989
2000
2015
7388
7403
mCksGesmCksmCdsAdsTdsmCdsmCdsTds
91
3228
|
GdsGdsmCdsGesmCesGksGk
|
|
1206997
2008
2023
7396
7411
AksGesTksTdsmCdsmCdsTdsGdsmCdsGds
38
3761
|
mCdsmCdsAesTesmCksmCk
|
|
1207005
2017
2032
7405
7420
AksmCesTksTdsTdsAdsTdsTdsGdsAds
41
2848
|
GdsTdsTesmCesmCksTk
|
|
1207013
2031
2046
7419
7434
AksGesmCksAdsTdsTdsTdsTdsmCdsAds
106
3458
|
AdsAdsGesmCesAksmCk
|
|
1207032
147
162
514
529
AksmCksTdsGdsTdsGdsTdsGdsmCdsTds
51
3267
|
mCdsTesTesmCesAksGk
|
|
1207040
155
170
N/A
N/A
TksGksAdsGdsAdsAdsmCdsGdsAdsmCds
71
3875
|
TdsGesTesGesTksGk
|
|
1207048
212
227
3563
3578
TkSGkSTdsGdsGdsTdsAdsmCdsAdsGds
63
2734
|
mCdsTesGesmCesmCksCk
|
|
1207056
220
235
3571
3586
GksGksTdsAdsmCdsAdsTdsTdsTdsGds
66
3269
|
TdsGesGesTesAksmCk
|
|
1207064
1780
1795
7168
7183
GksmCksmCdsTdsTdsdsmCdsAdsdsdsGds
58
3451
|
TesGesAeGksmCk
|
|
1207085
1788
1803
7176
7191
mCksTksGdsAdsTdsGdsAdsTdsGdsmCds
61
3679
|
mCdsTesTesGesmCksAk
|
|
1207093
1894
1909
7282
7297
mCksAksmCdsTdsGdsmCdsGdsGdsAdsAds
56
3682
|
AdsAesTesmCesAksGk
|
|
1207101
1902
1917
7290
7305
AksAksAdsGdsAdsTdsGdsAdsGdsTds
48
2768
|
mCdsmCesmCesTesGksAk
|
|
1207109
1917
1932
7305
7320
AksAksTdsmCdsAdsmCdsmCdsAdsAdsGds
42
4956
|
GdsAesGesGesGksAk
|
|
1207117
1925
1940
7313
7328
mCksAkSmCdsTdsGdsmCdsGdsGdsAdsAds
56
3683
|
TdsmCesAesmCesmCksAk
|
|
1207133
1933
1948
7321
7336
mCksAksmCdsTdsmCdsTdsmCdsTdsmCds
33
4957
|
AdsmCdsTesGesmCesGksGk
|
|
1207146
2003
2018
7391
7406
mCksTksGdsmCdsGdsmCdsmCdsAdsTds
76
3381
|
mCdsmCdsTesGesGesmCksGk
|
|
1207154
2011
2026
7399
7414
TksTkSGdsAdsGdsTdsTdsmCdsmCdsTds
48
2463
|
GdsmCesGesmCesmCksAk
|
|
1207162
2020
2035
7408
7423
AksGksmCdsAdsmCdsTdsTdsTdsAdsTds
55
3076
|
TdsGesAesGesTksTk
|
|
1207170
142
157
509
524
gksTksGdsmCdsTdsmCdsTdsTdsmCdsAds
47
4958
|
GdsmCesTksTesTksGe
|
|
1207178
150
165
N/A
N/A
AksmCksGdsAdsmCdsTdsGdsTdsGdsTds
50
3495
|
GdsmCesTksmCesTksTe
|
|
1207186
158
173
N/A
N/A
mCksAksGdsTdSGdsAdSGdsAdsAdsmCds
78
2579
|
GdsAesmCksTesGksTe
|
|
1207194
215
230
3566
3581
AksTksTdsTdGdsTdsGdsGdsTdsAdsCds
70
2963
|
AesGksmCesTksGe
|
|
1207202
223
238
3574
3589
GksTksGdsGdsGdsTdsAdsmCdsAdsTds
36
3497
|
TdsTesGksTesGksGe
|
|
1207210
1783
1798
7171
7186
GksAksTdsGdsmCdsmCdsTdsTdsGdsmCds
39
4959
|
AdsGesGksGesTksGe
|
|
1207218
1791
1806
7179
7194
mCksAksGdsmCdsTdsGdsAdsTdsGdsAds
89
4960
|
TdsGesmCksmCesTksTe
|
|
1207226
1897
1912
7285
7300
TksGksAdsGdsTdsmCdsmCdsmCdsTdsGds
65
3910
|
AdsGesmCksAesAksTe
|
|
1207234
1905
1920
7293
7308
GksGksGdsAdsAdsAdsGdsAdsTdsGdsAds
68
2997
|
GesTksmCesmCksmCe
|
|
1207242
1920
1935
7308
7323
mCksGksGdsAdSAdSTdSmCdsAdsmCds
26
3303
|
mCdsAdsAesGksGesAksGe
|
|
1207250
1928
1943
7316
7331
TksmCksTdsmCdsAdsmCdsTdsGdsmCds
31
3911
|
GdsGdsAesAksTesmCksAe
|
|
1207258
1936
1951
7324
7339
AksGksmCdsmCdsAdsmCdsTdsmCdsTds
29
4961
|
mCdsTdsmCesAksmCesTksGe
|
|
1207266
2006
2021
7394
7409
TksTksmCdsmCdsTdsGdsmCdsGdsmCds
70
3609
|
mCdsAdsTesmCksmCesTksGe
|
|
1207274
2014
2029
7402
7417
TksTksAdsTdTdGdsAdsGdsTdsTdsmCds
54
2694
|
mCesTksGesmCksGe
|
|
1207282
2023
2038
7411
7426
mCksAksAdsAdsGdsmCdsAdsmCdsTdsTds
68
3306
|
TdsAesTksTesGksAe
|
|
1207290
145
160
512
527
TksGksTdsGdsTdsGdsmCdsTdsmCdsTds
74
3112
|
TdsmCksAesGksmCesTk
|
|
1207298
153
168
N/A
N/A
AksGksAdsAdsmCdsGdsAdsmCdsTdsGds
52
3723
|
TdsGksTesGksmCesTk
|
|
1207306
210
225
3561
3576
TksGksGdsTdsAdsmCdsAdsGdsmCdsTds
91
2657
|
GdsmCksmCesGksGesTk
|
|
1207314
218
233
3569
3584
TksAksmCdsAdsTdsTdsTdsGdsTdsGds
66
2426
|
GdsTksAesCksAesGk
|
|
1207322
1778
1793
7166
7181
mCksTksTdsGdsmCdsAdsGdsGdsGdsTds
59
4962
|
GdsAksGesmCksmCesGk
|
|
1207330
1786
1801
7174
7189
GksAksTdsGdsAdsTdGdsmCdsmCdsTds
53
4963
|
TdsGksmCesAksGesGk
|
|
1207338
1892
1907
7280
7295
mCksmCksmCdsTdsGdsAdsGdsmCdsAds
74
3530
|
AdsTdsmCksAesGksGesAk
|
|
1207346
1900
1915
7288
7303
AksGksAdsTdsGdsAdGdsTdsmCdsmCds
76
2614
|
mCdsTksGesAksGesmCk
|
|
1207354
1915
1930
7303
7318
TksmCksAdsmCdsmCdsAdsAdsGdsGdsAds
44
4948
|
GdsGksGesAksAesAk
|
|
1207362
1923
1938
7311
7326
mCksTksGdsmCdsGdsGdsAdsAdsTdsmCds
46
3531
|
AdsmCksmCesAksAesGk
|
|
1207370
1931
1946
7319
7334
mCksTksmCdsTdsmCdsTdsmCdsAdsmCds
41
2461
|
TdsGdsmCksGesGksAesAk
|
|
1207378
2001
2016
7389
7404
GksmCksGdsmCdsmCdsAdsTdsmCdsmCds
80
4964
|
TdsGdsGksmCesGksmCesGk
|
|
1207386
2009
2024
7397
7412
GksAksGdsTdsTdsmCdsmCdsTdsGdsmCds
71
3837
|
GdsmCksmCesAksTesmCk
|
|
1207394
2018
2033
7406
7421
mCksAksmCdsTdsTdsTdsAdsTdsTdsGds
47
2924
|
AdsGksTesTksmCesmCk
|
|
1207402
2033
2048
7421
7436
TksmCksAdsGdsCdsAdsTdsTdsTdsTds
32
3534
|
mCdsAksAesAksGesmCk
|
|
1207410
148
163
515
530
GksAksmCksTdsGdsTdsGdsTdsGdsmCds
64
3343
|
TdsmCdsTksTksmCksAe
|
|
1207418
156
171
N/A
N/A
GksTksGksAdsGdsAdsAdsmCdsGdsAds
88
2424
|
mCdsTdsGksTksGksTe
|
|
1207426
213
228
3564
3579
TksTksGksTdsGdsGdsTdsAdsmCdsAds
54
2.811
|
GdsmCdsTksGksmCksmCe
|
|
1207434
221
236
3572
3587
GksGksGksTdsAdsmCdsAdsTdsTdsTds
63
3345
|
GdsTdsGksGksTksAe
|
|
1207442
1781
1796
7169
7184
TksksmCksmCdsTdsTdsdsmCdsAdsGds
70
3527
|
GdsGdsTksGksAksGe
|
|
1207450
1789
1804
7177
7192
GksmCksTksGdsAdsTdsGdsAdsTdsGds
67
3755
|
mCdsmCdsTksTksGksmCe
|
|
1207458
1895
1910
7283
7298
AksGksTksmCdsmCdsmCdsTdsGdsAdsGds
51
3758
|
mCdsAdsAksTksCksAe
|
|
1207466
1903
1918
7291
7306
GksAksAksAdsGdsAdsTdsGdsAdsGdsTds
36
2845
|
mCdsmCksmCksTksGe
|
|
1207474
1918
1933
7306
7321
GksAksAksTdsmCdsAdsmCdsmCdsAdsAds
36
4965
|
GdsGdsAksGksGksGe
|
|
1207482
1926
1941
7314
7329
TksmCksAksmCdsTdsGdsmCdsGdsGdsAds
50
3759
|
AdsTdsmCksAksmCkmCe
|
|
1207490
1934
1949
7322
7337
mCksmCksAksmCdsTdsmCdsTdsmCdsTds
38
4949
|
mCdsAdsmCdsTksGksGksmCe
|
|
1207498
2004
2019
7392
7407
mCksmCksTksGdsmCdsGdsmCdsmCdsAds
54
3457
|
TdsmCdsmCdsTksGksGksmCe
|
|
1207506
2012
2027
7400
7415
AksTksTksGdsAdsGdsTdsTdsmCdsmCds
66
2540
|
TdsGdsmCksGksmCksmCe
|
|
1207514
2021
2036
7409
7424
AksAksGksmCdsAdsmCdsTdsTdsTdsAds
42
3147
|
TdsTdsGksAksGksTe
|
|
TABLE 79
|
|
Reduction of FXII RNA (Huh7, electroporation,
|
4000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
Sequence and Chemistry
|
Compound
Start
Stop
Start
Stop
notation
FXII
SEQ
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
ID NO
|
|
1129531
2009
2024
7397
7412
GksAksGksTdsTdsmCdsmCdsTdsGdsmCds
21
3837
|
GdsmCdsmCdsAksTksmCk
|
|
1206475
142
157
509
524
GksTksGksmCdsTdsmCdsTdsTdsmCdsAdsG
32
4958
|
ds
mCdsTdsTksTksGk
|
|
1206487
1916
1931
7304
7319
AksTksmCksAdsmCdsmCdsAdsAdsGdsGds
33
4966
|
AdsGdsGdsGksAksAk
|
|
1206492
1935
1950
7323
7338
GksmCksmCksAdsmCdsTdsmCdsTdsmCdsTd
5
4967
|
s
mCdsAdsmCdsTksGksmCk
|
|
1206500
1783
1798
7171
7186
GksAksTksGdsmCdsmCdsTdsTdsGdsmCds
22
4959
|
AdsGdsGdsGksTksGk
|
|
1206902
145
160
512
527
TksGesTksGdsTdsGdsmCdsTdsmCdsTdsTds
33
3112
|
CdsAesGesCksTk
|
|
1206910
153
168
N/A
N/A
AksGesAksAdsCdsGdsAdsCdsTdsGdsT
39
3723
|
dsGdsTesGesmCksTk
|
|
1206918
210
225
3561
3576
TksGesGksTdsAdsmCdsAdsGdsmCdsTdsGd
45
2657
|
s
mCdsmCesGesGksTk
|
|
1206926
218
233
3569
3584
TksAesmCksAdsTdsTdsTdsGdsTdsGdsGds
17
2426
|
TdsAesmCesAksGk
|
|
1206934
1778
1793
7166
7181
mCksTesTksGdsmCdsAdsGdsGdsGdsTdsGd
31
4962
|
sAdsGesmCesmCksGk
|
|
1206942
1786
1801
7174
7189
GksAesTksGdsAdsTdsGdsmCdsmCdsTdsTd
24
4963
|
sGdsmCesAesGksGk
|
|
1206950
1892
1907
7280
7295
mCksmCesmCksTdsGdsAdsGdsmCdsAdsAds
47
3530
|
TdsCdsAesGesGksAk
|
|
1206958
1900
1915
7288
7303
AksGesAksTdsGdsAdsGdsTdsCdsCds
24
2614
|
CdsTdsGesAesGksmCk
|
|
1206966
1915
1930
7303
7318
TksmCesAksmCdsmCdsAdsAdsGdsGdsAds
24
4948
|
GdsGdsGesAesAksAk
|
|
1206974
1923
1938
7311
7326
mCksTesGksmCdsGdsGdsAdsAdsTdsmCds
15
3531
|
AdsmCdsmCesAesAksGk
|
|
1206982
1931
1946
7319
7334
CksTesCksTdsCdsTdsCdsAdsCdsTd
17
2461
|
sGdsmCdsGesGesAksAk
|
|
1206990
2001
2016
7389
7404
GksmCesGksmCdsmCdsAdsTdsmCdsmCdsT
73
4964
|
dsGdsGdsmCesGesmCksGk
|
|
1206998
2009
2024
7397
7412
GksAesGksTdsTdsmCdsmCdsTdsGdsmCds
21
3837
|
GdsmCdsmCesAesTksmCk
|
|
1207006
2018
2033
7406
7421
CksAesCksTdsTdsTdsAdsTdsTdsGdsAds
20
2924
|
GdsTesTesmCksmCk
|
|
1207014
2033
2048
7421
7436
TksmCesAksGdsmCdsAdsTdsTdsTdsTdsmC
44
3534
|
dsAdsAesAesGksCk
|
|
1207033
148
163
515
530
GksAksCdsTdsGdsTdsGdsTdsGdsCdsTd
11
3343
|
sCesTesTesCksAk
|
|
1207041
156
171
N/A
N/A
GksTksGdsAdsGdsAdsAdsmCdsGdsAdsmCds
48
2424
|
TesGesTesGksTk
|
|
1207049
213
228
3564
3579
TksTksGdsTdsGdsGdsTdsAdsmCdsAdsGds
25
2811
|
mCesTesGesmCksmCk
|
|
1207057
221
236
3572
3587
GksGksGdsTdsAdsmCdsAdsTdsTdsTdsGds
47
3345
|
TesGesGesTksAk
|
|
1207071
1781
1796
7169
7184
TksGksmCdsmCdsTdsTdsGdsmCdsAdsGds
35
3527
|
GdsGesTesGesAksGk
|
|
1207086
1789
1804
7177
7192
GksmCksTdsGdsAdsTdsGdsAdsTdsGdsmCd
24
3755
|
sCesTesTesGksCk
|
|
1207094
1895
1910
7283
7298
AksGksTdsmCdsmCdsmCdsTdsGdsAdsGdsm
36
3758
|
CdsAesAesTesTksAk
|
|
1207102
1903
1918
7291
7306
GksAksAdsAdsGdsAdsTdsGdsAdsGdsTds
28
2845
|
CesmCesmCesTksGk
|
|
1207110
1918
1933
7306
7321
GksAksAdsTdsCdsAdsCdsCdsAdsAds
24
4965
|
GdsGesAesGesGksGk
|
|
1207118
1926
1941
7314
7329
TksmCksAdsmCdsTdsGdsmCdsGdsGdsAds
12
3759
|
AdsTesmCesAesmCksmCk
|
|
1207135
1934
1949
7322
7337
mCksmksAdsCdsAdsCdsAdsCdsAds
16
4949
|
CdsAdsmCesTesGesmCksGk
|
|
1207155
2012
2027
7400
7415
mCksmCksAdsmCdsTdsGdsmCdsAdsT
33
2540
|
dsCdsCesTesGesGksCk
|
|
1207163
2021
2036
7409
7424
AksTksTdsGdsAdsGdsTdsTdsCdsmCdsTds
22
3147
|
GesmCesGesmCksmCk
|
|
1207171
143
158
510
525
AksAksGdsCdsAdsCdsTdsTdsTdsAdsTd
49
3037
|
sTesGesAesGksTk
|
|
1207179
151
166
N/A
N/A
TksGksTdsGdsmCdsTdsmCdsTdsTdsmCdsA
43
3571
|
dsGesmCksTesTksTe
|
|
1207187
208
223
3559
3574
AksAksCdsGdsAdsCdsTdsGdsTdsGdsTd
45
4968
|
sGesmCksTesmCksTe
|
|
1207195
216
231
3567
3582
GksTksAdsmCdsAdsGdsmCdsTdsGdsmCdsm
26
3039
|
CdsGesGksTesGksGe
|
|
1207203
1776
1791
7164
7179
mCksAksTdsTdsTdsGdsTdsGdsGdsTdsAds
68
3299
|
CesAksGesCksTe
|
|
1207211
1784
1799
7172
7187
TksGksCdsAdsGdsGdsGdsTdsGdsAdsGds
46
4969
|
mCesmCksGesGksmCe
|
|
1207219
1792
1807
7180
7195
TksGksAdsTdsGdsmCdsmCdsTdsTdsGdsmC
39
4970
|
dsAesGksGesGksTe
|
|
1207227
1898
1913
7286
7301
mCksmCksAdsGdsmCdsTdsGdsAdsTdsGds
31
2460
|
AdsTesGksmCesmCksTe
|
|
1207235
1906
1921
7294
7309
AksTksGdsAdsGdsTdsCdsCdsCdsTds
46
3073
|
GdsAesGksmCesAksAe
|
|
1207243
1921
1936
7309
7324
AksGksGdsGdsAdsAdsAdsGdsAdsTdsGds
26
3379
|
AesGksTesmCksmCe
|
|
1207251
1929
1944
7317
7332
GksmCksGdsGdsAdsAdsTdsmCdsAdsmCds
11
4971
|
CdsAesAksGesGksAe
|
|
1207259
1937
1952
7325
7340
mCksTksmCdsTdsmCdsAdsmCdsTdsGdsmCd
20
4972
|
sGdsGesAksAesTksmCe
|
|
1207267
2007
2022
7395
7410
mCksAksGdsmCdsmCdsAdsmCdsTdsmCdsT
32
3685
|
dsCdsTesCksAesCksTe
|
|
1207275
2016
2031
7404
7419
GksTksTdsmCdsmCdsTdsGdsmCdsGdsmCds
10
2771
|
mCdsAesTksmCesmCksTe
|
|
1207283
2024
2039
7412
7427
CksTksTdsTdsAdsTdsTdsGdsAdsGdsTds
24
3382
|
TesmCksmCesTksGe
|
|
1207291
146
161
513
528
TksmCksAdsAdsAdsGdsmCdsAdsmCdsTds
29
3190
|
TdsTesAksTesTksGe
|
|
1207299
154
169
N/A
N/A
mCksTksGdsTdsGdsTdsGdsmCdsTdsmCdsT
56
3799
|
dsTksmCesAksGesmCk
|
|
1207307
211
226
3562
3577
GksAksGdsAdsAdsmCdsGdsAdsmCdsTdsG
36
4973
|
dsTksGesTksGesmmCk
|
|
1207315
219
234
3570
3585
GksTksGdsGdsTdsAdsmCdsAdsGdsmCdsTd
88
3192
|
sGksCesCksGesGk
|
|
1207323
1779
1794
7167
7182
GksTksAdsmCdsAdsTdsTdsTdsGdsTdsGds
41
4974
|
GksTesAksmCesAk
|
|
1207331
1787
1802
7175
7190
mCksmCksTdsTdsGdsmCdsAdsGdsGdsGds
35
4975
|
TdsGksAesGksmCesmCk
|
|
1207339
1893
1908
7281
7296
TksmCksmCdsmCdsTdsGdsAdsGdsmCdsAds
44
3606
|
AdsTksmCesAksGesGk
|
|
1207347
1901
1916
7289
7304
AksAksGdsAdsTdsGdsAdsGdsTdsmCdsmC
37
2691
|
ds
mCksTesGksAesGk
|
|
1207355
1916
1931
7304
7319
AksTksmCdsAdsmCdsmCdsAdsAdsGdsGds
41
4966
|
AdsGksGesGksAesAk
|
|
1207363
1924
1939
7312
7327
AksmCksTdsGdsmCdsGdsGdsAdsAdsTdsm
22
3607
|
CdsAksmCesmCksAesAk
|
|
1207371
1932
1947
7320
7335
AksmCksTdsmCdsTdsmCdsTdsmCdsAdsmCd
22
4976
|
sTdsGksmCesGksGesAk
|
|
1207379
2002
2017
7390
7405
TksGksmCdsGdsmCdsmCdsAdsAdsCdsC
96
3305
|
dsTdsGksGesmCksGesmCk
|
|
1207387
2010
2025
7398
7413
TksGksAdsGdsTdsTdsmCdsmCdsTdsGdsmC
24
3913
|
dsGksTesTksAesTk
|
|
1207395
2019
2034
7407
7422
GksmCksAdsmCdsTdsTdsTdsAdsTdsTdsGds
26
3000
|
AksGesTksTesmCk
|
|
1207403
141
156
508
523
TksGksCksTdsmCdsTdsTdsmCdsAdsGds
38
4950
|
mCdsTdsTksTksGksTe
|
|
1207411
149
164
516
531
mCksGksAksmCdsTdsGdsTdsGdsTdsGdsm
24
3419
|
CdsTdsmCksTksTksmCe
|
|
1207419
157
172
N/A
N/A
AksGksTksGdsAdsGdsAdsAdsmCdsGdsAds
25
2502
|
mCdsTksGksTksGe
|
|
1207427
214
229
3565
3580
TksTksTksGdsTdsGdsGdsTdsAdsmCdsAds
31
2887
|
GdsTksTksGTCe
|
|
1207435
222
237
3573
3588
TksGksGksGdsTdsAdsmCdsAdsTdsTdsTds
53
3421
|
GdsTksGksGksTe
|
|
1207443
1782
1797
7170
7185
AksTksGksmCdsmCdsTdsTdsGdsmCdsAds
46
4947
|
GdsGdsGksTksGksAe
|
|
1207451
1790
1805
7178
7193
AksGksmCksTdsGdsAdsTdsGdsAdsTdsGds
49
3831
|
mCdsTksTksTksGe
|
|
1207459
1896
1911
7284
7299
GksAksGksTdsmCdsmCdsmCdsTdsGdsAds
33
3834
|
GdsmCdsAksAksTksmCe
|
|
1207467
1904
1919
7292
7307
GksGksAksAdsAdsGdsAdsTdsGdsAdsGds
38
2921
|
TdsmCksmCksmCksTe
|
|
1207475
1919
1934
7307
7322
GksGksAksAdsTdsmCdsAdsmCdsmCdsAds
16
3226
|
AdsGdsGksAksGksGe
|
|
1207483
1927
1942
7315
7330
mCksTksmCksAdsmCdsTdsGdsmCdsGdsGds
22
3835
|
AdsAdsTksmCksAksmCe
|
|
1207491
1935
1950
7323
7338
GksmCksmCksAdsmCdsTdsmCdsTdsmCdsTd
22
4967
|
s
mCdsAdsmCksTksGksmCe
|
|
1207499
2005
2020
7393
7408
TksmCksmCksTdsGdsmCdsGdsmCdsmCdsA
21
3533
|
dsTdsmCdsmCksTksGksGe
|
|
1207507
2013
2028
7401
7416
TksAksTksTdsGdsAdsGdsTdsTdsmCdsmCds
26
2617
|
TdsGksmCksGksmCe
|
|
1207515
2022
2037
7410
7425
AksAksAksGdsmCdsAdsmCdsTdsTdsTdsAd
21
3229
|
sTdsTksGksAksGe
|
|
TABLE 80
|
|
Reduction of FXII RNA (Huh7, electroporation, 4000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
|
Compound
Start
Stop
Start
Stop
Sequence and Chemistry notation
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1129531
2009
2024
7397
7412
GksAksGksTdsTdsmCdsmCdsTdsGdsmCds
19
3837
|
GdsmCdsmCdsAksTksmCk
|
|
1206476
144
159
511
526
GksTksGksTdsGdsmCdsTdsmCdsTdsTdsmC
24
4951
|
dsAdsGdsCksTksTk
|
|
1206481
1784
1799
7172
7187
TksGksAksTdsGdsCdsCdsTdsTdsGdsC
42
4969
|
dsAdsGdsGksGksTk
|
|
1206505
1917
1932
7305
7320
AksAksTksmCdsAdsmCdsmCdsAdsAdsGds
27
4956
|
GdsAdsGdsGksGksAk
|
|
1206508
1936
1951
7324
7339
AksGksmCksmCdsAdsmCdsTdsmCdsTdsmC
9
4961
|
dsTdsmCdsAdsmCksTksGk
|
|
1206903
146
161
513
528
mCksTesGksTdsGdsTdsGdsmCdsTdsmCdsT
14
3190
|
dsTdsmCesAesGksmCk
|
|
1206911
154
169
N/A
N/A
GksAesGksAdsAdsmCdsGdsAdsmCdsTdsG
29
3799
|
dsTdsGesTesGksmCk
|
|
1206919
211
226
3562
3577
GksTesGksGdsTdsAdsmCdsAdsGdsmCdsTd
24
4973
|
sGdsCesCesGksGk
|
|
1206927
219
234
3570
3585
GksTesAksmCdsAdsTdsTdsTdsGdsTdsGds
27
3192
|
GdsTesAesmCksAk
|
|
1206935
1779
1794
7167
7182
mCksmCesTksTdsGdsmCdsAdsGdsGdsGds
51
4974
|
TdsGdsAesGesmCksmCk
|
|
1206943
1787
1802
7175
7190
TksGesAksTdsGdsAdsTdsGdsmCdsmCdsTd
36
4975
|
sTdsGesCesAksGk
|
|
1206951
1893
1908
7281
7296
TksCesmCksCdsTdsGdsAdsGdsCdsAds
46
3606
|
AdsTdsmCesAesGksGk
|
|
1206959
1901
1916
7289
7304
AksAesGksAdsTdsGdsAdsGdsTdsmCdsmC
34
2691
|
ds
mCdsTesGesAksGk
|
|
1206967
1916
1931
7304
7319
AksTesmCksAdsmCdsmCdsAdsAdsGdsGds
14
4966
|
AdsGdsGesGesAksAk
|
|
1206975
1924
1939
7312
7327
AksmCesTksGdsmCdsGdsGdsAdsAdsTdsm
13
3607
|
CdsAdsmCesmCesAksAk
|
|
1206983
1932
1947
7320
7335
AksmCesTksmCdsTdsmCdsTdsmCdsAdsCds
9
4976
|
TdsGdsCesGesGksAk
|
|
1206991
2002
2017
7390
7405
TksGesmCksGdsmCdsmCdsAdsTdsmCdsmC
64
3305
|
dsTdsGdsGesCesGksCk
|
|
1206999
2010
2025
7398
7413
TksGesAksGdsTdsTdsmCdsmCdsTdsGdsmC
12
3913
|
dsGdsmCesmCesAksTk
|
|
1207007
2019
2034
7407
7422
GksmCesAksmCdsTdsTdsTdsAdsTdsTdsGds
7
3000
|
AdsGesTesTksmCk
|
|
1207026
141
156
508
523
TksGksCdsTdsCdsTdsTdsCdsAdsGds
49
4950
|
CdsTesTesTesGksTk
|
|
1207034
149
164
516
531
mCksGksAdsmCdsTdsGdsTdsGdsTdsGdsm
36
3419
|
CdsTesmCesTesTksmCk
|
|
1207042
157
172
N/A
N/A
AksGksTdsGdsAdsGdsAdsAdsmCdsGdsAds
42
2502
|
mCesTesGesTksGk
|
|
1207050
214
229
3565
3580
TksTksTdsGdsTdsGdsGdsTdsAdsmCdsAds
43
2887
|
GesmCesTesGksmCk
|
|
1207058
222
237
3573
3588
TksGksGdsGdsTdsAdsmCdsAdsTdsTdsTds
25
3421
|
GesTesGesGksTk
|
|
1207078
1782
1797
7170
7185
AksTksGdsmCdsmCdsTdsTdsGdsmCdsAds
52
4947
|
GdsGesGesTesGksAk
|
|
1207087
1790
1805
7178
7193
AksGksCdsTdsGdsAdsTdsGdsAdsTdsGds
42
3831
|
mCesmCes
|
|
1207095
1896
1911
7284
7299
GksAksGdsTdsmCdsmCdsmCdsTdsGdsAds
25
3834
|
GdsmCesAesAesTksmCk
|
|
1207103
1904
1919
7292
7307
GksGksAdsAdsAdsGdsAdsTdsGdsAdsGds
31
2921
|
TesmCesmCesmCksTk
|
|
1207111
1919
1934
7307
7322
GksGksAdsAdsTdsCdsAdsCdsCdsAds
9
3226
|
AdsGesGesAesGksGk
|
|
1207120
1927
1942
7315
7330
mCksTksmCdsAdsmCdsTdsGdsmCdsGdsGds
10
3835
|
AdsAesTesCesAksCk
|
|
1207137
1935
1950
7323
7338
GksmCksmCdsAdsmCdsTdsmCdsTdsmCdsTd
19
4967
|
sCdsAesCesTesGksCk
|
|
1207148
2005
2020
7393
7408
TksmCksmCdsTdsGdsmCdsGdsmCdsmCdsA
13
3533
|
dsTdsmCesmCesTesGksGk
|
|
1207156
2013
2028
7401
7416
TksAksTdsTdsGdsAdsGdsTdsTdsmCdsmCds
9
2617
|
TesGesmCesGksmCk
|
|
1207164
2022
2037
7410
7425
AksAksAdsGdsCdsAdsCdsTdsTdsTdsAd
5
3229
|
sTesTesGesAksGk
|
|
1207172
144
159
511
526
GksTksGdsTdsGdsmCdsTdsmCdsTdsTdsmC
14
4951
|
dsAesGksmCesTksTe
|
|
1207180
152
167
N/A
N/A
GksAksAdsmCdsGdsAdsmCdsTdsGdsTdsG
32
3647
|
dsTesGksmCesTksmCe
|
|
1207188
209
224
3560
3575
GksGksTdsAdsmCdsAdsGdsmCdsTdsGdsm
23
4952
|
CdsmCesGksGesTksGe
|
|
1207196
217
232
3568
3583
AksCksAdsTdsTdsTdsGdsTdsGdsGdsTds
13
3114
|
AesmCksAesGksmCe
|
|
1207204
1777
1792
7165
7180
TksTksGdsmCdsAdsGdsGdsGdsTdsGdsAds
50
3375
|
GesCksCesGksGe
|
|
1207212
1785
1800
7173
7188
AksTksGdsAdsTdsGdsCdsCdsTdsTdsGd
25
3603
|
sCesAksGesGksGe
|
|
1207220
1793
1808
7181
7196
CksCksCdsAdsGdsmCdsTdsGdsAdsTds
26
4953
|
GdsAesTksGesmCksmCe
|
|
1207228
1899
1914
7287
7302
GksAksTdsGdsAdsGdsTdsmCdsmCdsmCds
6
2537
|
TdsGesAksGesmCksAe
|
|
1207236
1914
1929
7302
7317
mCksAksmCdsmCdsAdsAdsGdsGdsAdsGds
12
4954
|
GdsGesAksAesAksGe
|
|
1207244
1922
1937
7310
7325
TksGksmCdsGdsGdsAdsAdsTdsmCdsAdsm
17
3455
|
CdsmCesAksAesGksGe
|
|
1207252
1930
1945
7318
7333
TksmCksTdsmCdsTdsmCdsAdsmCdsTdsGds
16
4955
|
mCdsGesGksAesAksTe
|
|
1207260
2000
2015
7388
7403
mCksGksmCdsmCdsAdsTdsmCdsmCdsTdsG
77
3228
|
dsGdsmCesGksmCesGksGe
|
|
1207268
2008
2023
7396
7411
AksGksTdsTdsmCdsmCdsTdsGdsmCdsGdsm
18
3761
|
CdsmCesAksTesmCksmCe
|
|
1207276
2017
2032
7405
7420
AksCksTdsTdsTdsAdsTdsTdsGdsAdsGds
8
2848
|
TesTksmCesmCksTe
|
|
1207284
2031
2046
7419
7434
AksGksmCdsAdsTdsTdsTdsTdsmCdsAdsAd
20
3458
|
sAesGksmCesAksmCe
|
|
1207292
147
162
514
529
AksmCksTdsGdsTdsGdsTdsGdsmCdsTdsmC
36
3267
|
dsTksTesmCksAesGk
|
|
1207300
155
170
N/A
N/A
TksGksAdsGdsAdsAdsmCdsGdsAdsmCdsT
25
3875
|
dsGksTesGksTesGk
|
|
1207308
212
227
3563
3578
TksGksTdsGdsGdsTdsAdsCdsAdsGdsCd
18
2734
|
sTksGesmCksmCesGk
|
|
1207316
220
235
3571
3586
GksGksTdsAdsCdsAdsTdsTdsTdsGdsTds
49
3269
|
GksGesTksAesmCk
|
|
1207324
1780
1795
7168
7183
GksmCksmCdsTdsTdsGdsmCdsAdsGdsGds
47
3451
|
GdsTksGesAksGesmCk
|
|
1207332
1788
1803
7176
7191
CksTksGdsAdsTdsGdsAdsTdsGdsCds
29
3679
|
CdsTksTesGksmCesAk
|
|
1207340
1894
1909
7282
7297
GksTksmCdsmCdsmCdsTdsGdsAdsGdsmCds
57
3682
|
AdsAksTesCksAesGk
|
|
1207348
1902
1917
7290
7305
AksAksAdsGdsAdsTdsGdsAdsGdsTdsCds
32
2768
|
CksCesTksGesAk
|
|
1207356
1917
1932
7305
7320
AksAksTdsmCdsAdsmCdsmCdsAdsAdsGds
57
4956
|
GdsAksGesGksGesAk
|
|
1207364
1925
1940
7313
7328
mCksAksmCdsTdsGdsmCdsGdsGdsAdsAds
14
3683
|
TdsmCksAesmCksmCesAk
|
|
1207372
1933
1948
7321
7336
CksAksCdsTdsCdsTdsCdsTdsCdsAd
20
4957
|
s
mCdsTksGesmCksGesGk
|
|
1207380
2003
2018
7391
7406
CksTksGdsCdsGdsmCdsCdsAdsTdsC
50
3381
|
ds
mCdsTksGesGksmCesGk
|
|
1207388
2011
2026
7399
7414
TksTksGdsAdsGdsTdsTdsmCdsmCdsTdsGds
27
2463
|
mCksGesmCksmCesAk
|
|
1207396
2020
2035
7408
7423
AksGksmCdsAdsmCdsTdsTdsTdsAdsTdsTds
17
3076
|
GksAesGksTesTk
|
|
1207404
142
157
509
524
GksTksGksmCdsTdsmCdsTdsTdsmCdsAdsG
33
4958
|
ds
mCdsTksTksTksGe
|
|
1207412
150
165
N/A
N/A
AksmCksGksAdsmCdsTdsGdsTdsGdsTdsGd
24
3495
|
sCdsTksCksTksTe
|
|
1207420
158
173
N/A
N/A
CksAksGksTdsGdsAdsGdsAdsAdsCdsG
20
2579
|
dsAdsCksTfesGksTe
|
|
1207428
215
230
3566
3581
AksTksTksTdsGdsTdsGdsGdsTdsAdsCds
27
2963
|
AdsGksmCksTksGe
|
|
1207436
223
238
3574
3589
GksTksGksGdsGdsTdsAdsmCdsAdsTdsTds
25
3497
|
TdsGksTksGksGe
|
|
1207444
1783
1798
7171
7186
GksAksTksGdsCdsCdsTdsTdsGdsCds
43
4959
|
AdsGdsGksGksTksGe
|
|
1207452
1791
1806
7179
7194
mCksAksGksmCdsTdsGdsAdsTdsGdsAdsTds
24
4960
|
GdsmCksmCksTksTe
|
|
1207460
1897
1912
7285
7300
TksGksAksGdsTdsmCdsmCdsmCdsTdsGds
36
3910
|
AdsGdsCksAksAksTe
|
|
1207468
1905
1920
7293
7308
GksGksGksAdsAdsAdsGdsAdsTdsGdsAds
54
2997
|
GdsTksmCksmCksmCe
|
|
1207476
1920
1935
7308
7323
mCksGksGksAdsAdsTdsmCdsAdsmCdsmCd
22
3303
|
sAdsAdsGksGksAksGe
|
|
1207484
1928
1943
7316
7331
TksCksTksmCdsAdsCdsTdsGdsCdsGds
9
3911
|
GdsAdsAksTksCksAe
|
|
1207492
1936
1951
7324
7339
AksGksmCksmCdsAdsmCdsTdsmCdsTdsmC
40
4961
|
dsTdsmCdsAksmCksTksGe
|
|
1207500
2006
2021
7394
7409
TksTksmCksmCdsTdsGdsmCdsGdsmCdsmCd
33
3609
|
sAdsTesmCksTksTksGe
|
|
1207508
2014
2029
7402
7417
TksTksAksTdsTdsGdsAdsGdsTdsTdsmCdsm
17
2694
|
CdsTksGksmCksGe
|
|
1207516
2023
2038
7411
7426
mCksAksAksAdsGdsmCdsAdsmCdsTdsTds
5
3306
|
TdsAdsTksTksGksAe
|
|
TABLE 81
|
|
Reduction of FXII RNA (Huh7, electroporation, 4000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
Sequence and Chemistry notation
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1129531
2009
2024
7397
7412
GksAksGksTdsTdsmCdsmCdsTdsGdsmCdsGd
13
3837
|
s
mCdsmCdsAksTksmCk
|
|
1206496
208
223
3559
3574
GksTksAksmCdsAdsGdsmCdsTdsGdsmCdsm
27
4968
|
CdsGdsGdsTksGksGk
|
|
1206501
1786
1801
7174
7189
GksAksTksGdsAdsTdsGdsmCdsmCdsTdsTds
48
4963
|
GdsmCdsAksGksGk
|
|
1206506
1918
1933
7306
7321
GksAksAksTdsmCdsAdsmCdsmCdsAdsAdsG
10
4965
|
dsGdsAdsGksGksGk
|
|
1206509
1937
1952
7325
7340
mCksAksGksmCdsmCdsAdsmCdsTdsmCdsTds
6
4972
|
mCdsTdsmCdsAksmCksTk
|
|
1206904
147
162
514
529
AksmCesTksGdsTdsGdsTdsGdsmCdsTdsmCds
23
3267
|
TdsTesmCesAksGk
|
|
1206912
155
170
N/A
N/A
TksGesAksGdsAdsAdsmCdsGdsAdsmCdsTds
23
3875
|
GdsTesGesTksGk
|
|
1206920
212
227
3563
3578
TksGesTksGdsGdsTdsAdsmCdsAdsGdsmCds
13
2734
|
TdsGesmCesmCksGk
|
|
1206928
220
235
3571
3586
GksGesTksAdsmCdsAdsTdsTdsTdsGdsTdsG
45
3269
|
dsGesTesAksmCk
|
|
1206936
1780
1795
7168
7183
GksmCesmCksTdsTdsGdsmCdsAdsGdsGdsGd
48
3451
|
sTdsGesAEsGksmCk
|
|
1206944
1788
1803
7176
7191
mCksTesGksAdsTdsGdsAdsTdsGdsmCdsmCd
46
3679
|
sTdsTesGesmCksAk
|
|
1206952
1894
1909
7282
7297
GksTesmCksmCdsmCdsTdsGdsAdsGdsmCds
19
3682
|
AdsAdsTesmCesAksGk
|
|
1206960
1902
1917
7290
7305
AksAesAksGdsAdsTdsGdsAdsGdsTdsmCdsm
17
2768
|
CdsmCesTesGksAk
|
|
1206968
1917
1932
7305
7320
AksAesTksmCdsAdsmCdsmCdsAdsAdsGdsG
41
4956
|
dsAdsGesGesGksAk
|
|
1206976
1925
1940
7313
7328
mCksAesmCksTdsGdsmCdsGdsGdsAdsAdsTd
29
3683
|
s
mCdsAesmCesmCksAk
|
|
1206984
1933
1948
7321
7336
mCksAesmCksTdsmCdsTdsmCdsTdsmCdsAds
N.D
4957
|
mCdsTdsGesmCesGksGk
|
|
1206992
2003
2018
7391
7406
mCksTesGksmCdsGdsmCdsmCdsAdsTdsmCds
77
3381
|
mCdsTdsGesGesmCksGk
|
|
1207000
2011
2026
7399
7414
TksTesGksAdsGdsTdsTdsmCdsmCdsTdsGdsm
11
2463
|
CdsGesmCesmCksAk
|
|
1207008
2020
2035
7408
7423
AksGesmCksAdsmCdsTdsTdsTdsAdsTdsTds
5
3076
|
GdsAesGesTksTk
|
|
1207027
142
157
509
524
GksTksGdsmCdsTdsmCdsTdsTdsmCdsAdsGds
40
4958
|
mCesTesTesTksGk
|
|
1207035
150
165
N/A
N/A
AksmCksGdsAdsmCdsTdsGdsTdsGds
35
3495
|
TdsGdsmCesTesmCesTksTk
|
|
1207043
158
173
N/A
N/A
mCksAksGdsTdsGdsAdsGdsAdsAdsm
36
2579
|
CdsGdsAesmCesTesGksTk
|
|
1207051
215
230
3566
3581
AksTksTdsTdsGdsTdsGdsGdsTdsAd
43
2963
|
s
mCdsAesGesmCesTksGk
|
|
1207059
223
238
3574
3589
GksTksGdsGdsGdsTdsAdsmCdsAdsT
25
3497
|
dsTdsTesGesTesGksGk
|
|
1207079
1783
1798
7171
7186
GksAksTdsGdsmCdsmCdsTdsTdsGds
37
4959
|
mCdsAdsGesGesGesTksGk
|
|
1207088
1791
1806
7179
7194
mCksAksGdsmCdsTdsGdsAdsTdsGds
40
4960
|
AdsTdsGesmCesmCesTksTk
|
|
1207096
1897
1912
7285
7300
TksGksAdsGdsTdsmCdsmCdsmCdsTd
27
3910
|
sGdsAdsGesmCesAesAksTk
|
|
1207104
1905
1920
7293
7308
GksGksGdsAdsAdsAdsGdsAdsTdsGd
31
2997
|
sAdsGesTesmCesmCksmCk
|
|
1207112
1920
1935
7308
7323
mCksGksGdsAdsAdsTdsmCdsAdsmCd
19
3303
|
s
mCdsAdsAesGesGesAksGk
|
|
1207123
1928
1943
7316
7331
TksmCksTdsmCdsAdsmCdsTdsGdsmC
7
3911
|
dsGdsGdsAesAesTesmCksAk
|
|
1207139
1936
1951
7324
7339
AksGksmCdsmCdsAdsmCdsTdsmCdsT
7
4961
|
ds
mCdsTdsmCesAesmCesTksGk
|
|
1207149
2006
2021
7394
7409
TksTksmCdsmCdsTdsGdsmCdsGdsmC
13
3609
|
ds
mCdsAdsTEsmCesmCesTksGk
|
|
1207157
2014
2029
7402
7417
TksTksAdsTdsTdsGdsAdsGdsTdsTd
15
2694
|
s
mCdsmCesTesGesmCksGk
|
|
1207165
2023
2038
7411
7426
mCksAksAdsAdsGdsmCdsAdsmCdsTd
18
3306
|
sTdsTdsAesTesTesGksAk
|
|
1207173
145
160
512
527
TksGksTdsGdsTdsGdsmCdsTdsmCds
12
3112
|
TdsTdsmCesAksGesmCksTe
|
|
1207181
153
168
N/A
N/A
AksGksAdsAdsmCdsGdsAdsmCdsTds
53
3723
|
GdsTdsGesTksGesmCksTe
|
|
1207189
210
225
3561
3576
TksGksGdsTdsAdsmCdsAdsGdsmCds
30
2657
|
TdsGdsmCesmCksGesGksTe
|
|
1207197
218
233
3569
3584
TksAksmCdsAdsTdsTdsTdsGdsTdsG
14
2426
|
dsGdsTesAksmCesAksGe
|
|
1207205
1778
1793
7166
7181
mCksTksTdsGdsmCdsAdsGdsGdsGds
48
4962
|
TdsGdsAesGksmCesmCksGe
|
|
1207213
1786
1801
7174
7189
GksAksTdsGdsAdsTdsGdsmCdsmCds
16
4963
|
TdsTdsGesmCksAesGksGe
|
|
1207221
1892
1907
7280
7295
mCksmCksmCdsTdsGdsAdsGdsmCdsA
53
3530
|
dsAdsTdsmCesAksGesGksAe
|
|
1207229
1900
1915
7288
7303
AksGksAdsTdsGdsAdsGdsTdsmCdsm
14
2614
|
CdsmCdsTesGksAesGksmCe
|
|
1207237
1915
1930
7303
7318
TksmCksAdsmCdsmCdsAdsAdsGdsGd
30
4948
|
sAdsGdsGesGksAesAksAe
|
|
1207245
1923
1938
7311
7326
mCksTksGdsmCdsGdsGdsAdsAdsTds
11
3531
|
mCdsAdsmCesmCksAesAksGe
|
|
1207253
1931
1946
7319
7334
mCksTksmCdsTdsmCdsTdsmCdsAdsm
9
2461
|
CdsTdsGdsmCesGksGesAksAe
|
|
1207261
2001
2016
7389
7404
GksmCksGdsmCdsmCdsAdsTdsmCdsm
56
4964
|
CdsTdsGdsGesmCksGesmCksGe
|
|
1207269
2009
2024
7397
7412
GksAksGdsTdsTdsmCdsmCdsTdsGds
11
3837
|
mCdsGdsmCesmCksAesTksmCe
|
|
1207277
2018
2033
7406
7421
mCksAksmCdsTdsTdsTdsAdsTdsTds
6
2924
|
GdsAdsGesTksTesmCksmCe
|
|
1207285
2033
2048
7421
7436
TksmCksAdsGdsmCdsAdsTdsTdsTds
36
3534
|
TdsmCdsAesAksAesGmCe
|
|
1207293
148
163
515
530
GksAksmCdsTdsGdsTdsGdsTdsGdsm
12
3343
|
CdsTdsmCksTesTksmCesAk
|
|
1207301
156
171
N/A
N/A
GksTksGdsAdsGdsAdsAdsmCdsGdsA
29
2424
|
ds
mCdsTksGesTksGesTk
|
|
1207309
213
228
3564
3579
TksTksGdsTdsGdsGdsTdsAdsmCdsA
21
2811
|
dsGdsmCksTesGksmCesmCk
|
|
1207317
221
236
3572
3587
GksGksGdsTdsAdsmCdsAdsTdsTdsT
46
3345
|
dsGdsTksGesGksTesAk
|
|
1207325
1781
1796
7169
7184
TksGksmCdsmCdsTdsTdsGdsmCdsAd
28
3527
|
sGdsGdsGksTesGksAesGk
|
|
1207333
1789
1804
7177
7192
GksmCksTdsGdsAdsTdsGdsAdsTdsG
26
3755
|
ds
mCdsmCksTesTksGesmCk
|
|
1207341
1895
1910
7283
7298
AksGksTdsmCdsmCdsmCdsTdsGdsAd
27
3758
|
sGdsmCdsAksAesTksmCesAk
|
|
1207349
1903
1918
7291
7306
GksAksAdsAdsGdsAdsTdsGdsAdsGd
29
2845
|
sTdsmCksmCesmCksTesGk
|
|
1207357
1918
1933
7306
7321
GksAksAdsTdsmCdsAdsmCdsmCdsAd
35
4965
|
sAdsGdsGksAesGksGesGk
|
|
1207365
1926
1941
7314
7329
TksmCksAdsmCdsTdsGdsmCdsGdsGd
10
3759
|
sAdsAdsTksmCesAksmCesmCk
|
|
1207373
1934
1949
7322
7337
mCksmCksAdsmCdsTdsmCdsTdsmCds
23
4949
|
TdsmCdsAdsmCksTesGksmCesGk
|
|
1207381
2004
2019
7392
7407
mCksmCksTdsGdsmCdsGdsmCdsmCds
9
3457
|
AdsTdsmCdsmCksTesGksGesmCk
|
|
1207389
2012
2027
7400
7415
AksTksTdsGdsAdsGdsTdsTdsmCdsm
44
2540
|
CdsTdsGksmCesGksmCesmCk
|
|
1207397
2021
2036
7409
7424
AksAksGdsmCdsAdsmCdsTdsTdsTds
22
3147
|
AdsTdsTksGesAksGesTk
|
|
1207405
143
158
510
525
TksGksTksGdsmCdsTdsmCdsTdsTds
29
3037
|
mCdsAdsGdsmCksTksTksTe
|
|
1207413
151
166
N/A
N/A
AksAksmCksGdsAdsmCdsTdsGdsTds
24
3571
|
GdsTdsGdsmCksTksmCksTe
|
|
1207421
208
223
3559
3574
GksTksAksmCdsAdsGdsmCdsTdsGds
25
4968
|
mCdsmCdsGdsGksTksGksGe
|
|
1207429
216
231
3567
3582
mCksAksTksTdsTdsGdsTdsGdsGdsT
14
3039
|
dsAdsmCdsAksGksmCksTe
|
|
1207437
1776
1791
7164
7179
TksGksmCksAdsGdsGdsGdsTdsGdsA
37
3299
|
dsGdsmCdsmCksGksGksmCe
|
|
1207445
1784
1799
7172
7187
TksGksAksTdsGdsmCdsmCdsTdsTds
36
4969
|
GdsmCdsAdsGksGksGksTe
|
|
1207453
1792
1807
7180
7195
mCksmCksAksGdsmCdsTdsGdsAdsTd
28
4970
|
sGdsAdsTdsGksmCksmCksTe
|
|
1207461
1898
1913
7286
7301
AksTksGksAdsGdsTdsmCdsmCdsmCd
21
2460
|
sTdsGdsAdsGksmCksAksAe
|
|
1207469
1906
1921
7294
7309
AksGksGksGdsAdsAdsAdsGdsAdsTd
15
3073
|
sGdsAdsGksTksmCksmCe
|
|
1207477
1921
1936
7309
7324
GksmCksGksGdsAdsAdsTdsmCdsAds
15
3379
|
mCdsmCdsAdsAksGksGksAe
|
|
1207485
1929
1944
7317
7332
mCksTksmCksTdsmCdsAdsmCdsTdsG
12
4971
|
ds
mCdsGdsGdsAksAksTksmCe
|
|
1207493
1937
1952
7325
7340
mCksAksGksmCdsmCdsAdsmCdsTdsm
14
4972
|
CdsTdsmCdsTdsmCksAksmCksTe
|
|
1207501
2007
2022
7395
7410
GksTksTksmCdsmCdsTdsGdsmCdsGd
21
3685
|
s
mCdsmCdsAdsTksmCksmCksTe
|
|
1207509
2016
2031
7404
7419
mCksTksTksTdsAdsTdsTdsGdsAds
8
2771
|
GdsTdsTdsmCksmCksTksGe
|
|
1207517
2024
2039
7412
7427
TksmCksAksAdsAdsGdsmCdsAds
11
3382
|
mCdsTdsTdsTdsAksTksTksGe
|
|
TABLE 82
|
|
Reduction of FXII RNA (Huh7, electroporation, 4000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
SEQ
|
Compound
Start
Stop
Start
Stop
Sequence and Chemistry
FXII
ID
|
Number
Site
Site
Site
Site
notation (5′ to 3′)
(% UTC)
NO
|
|
1129531
2009
2024
7397
7412
GksAksGksTdsTdsCdsCdsTdsGdsCd
12
3837
|
sGdsmCdsmCdsAksTksmCk
|
|
1206479
209
224
3560
3575
GksGksTksAdsCdsAdsGdsmCdsTdsG
20
4952
|
dsCdsmCdsGdsGksTksGk
|
|
1206502
1787
1802
7175
7190
TksGksAksTdsGdsAdsTdsGdsmCdsm
27
4975
|
CdsTdsTdsGdsCksAksGk
|
|
1206507
1929
1944
7317
7332
mCksTksmCksTdsmCdsAdsCdsTdsGd
11
4971
|
s
mCdsGdsGdsAdsAksTksCk
|
|
1206897
2001
2016
7389
7404
GksmCksGksmCdsAdsTdsmCdsmCds
41
4964
|
TdsGdsGdsmCdsGksmCksGk
|
|
1206905
148
163
515
530
GksAesmCksTdsGdsTdsGdsTdsGdsm
10
3343
|
CdsTdsCdsTesTesCksAk
|
|
1206913
156
171
N/A
N/A
GksTesGksAdsGdsAdsAdsCdsGdsAd
21
2424
|
sCdsTdsGesTesGksTk
|
|
1206921
213
228
3564
3579
TksTesGksTdsGdsGdsTdsAdsCdsAd
21
2811
|
sGdsmCdsTesGesmCksmCk
|
|
1206929
221
236
3572
3587
GksGesGksTdsAdsCdsAdsTdsTdsTd
22
3345
|
sGdsTdsGesGesTksAk
|
|
1206937
1781
1796
7169
7184
TksGesCksCdsTdsTdsGdsCdsAdsGd
58
3527
|
sGdsGdsTesGesAksGk
|
|
1206945
1789
1804
7177
7192
GksmCesTksGdsAdsTdsGdsAdsTdsG
23
3755
|
ds
mCdsmCdsTesTesGksmCk
|
|
1206953
1895
1910
7283
7298
AksGesTksmCdsmCdsmCdsTdsGdsAd
8
3758
|
sGdsCdsAdsAesTesCksAk
|
|
1206961
1903
1918
7291
7306
GksAesAksAdsGdsAdsTdsGdsAdsGd
17
2845
|
sTdsmCdsmCesmCesTksGk
|
|
1206969
1918
1933
7306
7321
GksAesAksTdsmCdsAdsmCdsmCdsAd
18
4965
|
sAdsGdsGdsAesGesGksGk
|
|
1206977
1926
1941
7314
7329
TksCesAksCdsTdsGdsCdsGdsGdsAd
12
3759
|
sAdsTdsmCesAesmCksmCk
|
|
1206985
1934
1949
7322
7337
mCksmCesAksmCdsTdsmCdsTdsmCds
13
4949
|
TdsmCdsAdsmCdsTesGesmCksGk
|
|
1206993
2004
2019
7392
7407
mCksmCesTksGdsmCdsGdsmCdsmCds
7
3457
|
AdsTdsmCdsmCdsTesGesGksmCk
|
|
1207001
2012
2027
7400
7415
AksTesTksGdsAdsGdsTdsTdsmCdsm
28
2540
|
CdsTdsGdsTesGesTksTk
|
|
1207009
2021
2036
7409
7424
AksAesGksmCdsAdsmCdsTdsTdsTds
N.D
3147
|
AdsTdsTdsGesAesGksTk
|
|
1207028
143
158
510
525
TksGksTdsGdsmCdsTdsmCdsTdsTds
72
3037
|
mCdsAdsGesmCesTesTksTk
|
|
1207036
151
166
N/A
N/A
AksAksmCdsGdsAdsmCdsTdsGdsTdsG
19
3571
|
dsTdsGesmCesTesmCksTk
|
|
1207044
208
223
3559
3574
GksTksAdsmCdsAdsGdsmCdsTdsGdsm
58
4968
|
CdsmCdsGesGesTesGksGk
|
|
1207052
216
231
3567
3582
mCksAksTdsTdsTdsGdsTdsGdsGdsT
24
3039
|
dsAdsCesAesGesCksTk
|
|
1207060
1776
1791
7164
7179
TksGksmCdsAdsGdsGdsGdsTdsGdsAd
26
3299
|
sGdsmCesmCesGesGksmCk
|
|
1207081
1784
1799
7172
7187
TksGksAdsTdsGdsmCdsmCdsTdsTdsG
39
4969
|
ds
mCdsAesGesGesGksTk
|
|
1207089
1792
1807
7180
7195
mCksmCksAdsGdsmCdsTdsGdsAdsTds
45
4970
|
GdsAdsTesGesmCksTk
|
|
1207097
1898
1913
7286
7301
AksTksGdsAdsGdsTdsmCdsmCdsmCds
14
2460
|
TdsGdsAesGesmCesAksAk
|
|
1207105
1906
1921
7294
7309
AksGksGdsGdsAdsAdsAdsGdsAdsTds
25
3073
|
GdsAesGesTesmCksmCk
|
|
1207113
1921
1936
7309
7324
GksmCksGdsGdsAdsAdsTdsmCdsAdsm
8
3379
|
CdsCdsAesAesGesGksAk
|
|
1207125
1929
1944
7317
7332
mCksTksmCdsTdsmCdsAdsmCdsTdsGd
9
4971
|
s
mCdsGdsGesAesAesTksmCk
|
|
1207141
1937
1952
7325
7340
mCksAksGdsmCdsmCdsAdsmCdsTdsmC
28
4972
|
dsTdsmCdsTesmCesAesmCksTk
|
|
1207150
2007
2022
7395
7410
GksTksTdsmCdsmCdsTdsGdsmCdsGds
21
3685
|
mCdsmCdsAesTesmCesmCksTk
|
|
1207158
2016
2031
7404
7419
CksTksTdsTdsAdsTdsTdsGdsAdsGds
27
2771
|
TdsTesmCesmCesTksGk
|
|
1207166
2024
2039
7412
7427
TksmCksAdsAdsAdsGdsmCdsAdsmCds
22
3382
|
TdsTdsTesAesTesTksGk
|
|
1207174
146
161
513
528
mCksTksGdsTdsGdsTdsGdsmCdsTdsm
28
3190
|
CdsTdsTesmCksAesGksmCe
|
|
1207182
154
169
N/A
N/A
GksAksGdsAdsAdsmCdsGdsAdsmCdsT
16
3799
|
dsGdsTesGksTesGksmCe
|
|
1207190
211
226
3562
3577
GksTksGdsGdsTdsAdsmCdsAdsGdsmC
5
4973
|
dsTdsGesCksCesGksGe
|
|
1207198
219
234
3570
3585
GksTksAdsCdsAdsTdsTdsTdsGdsTds
48
3192
|
GdsGesTksAesmCksAe
|
|
1207206
1779
1794
7167
7182
mCksmCksTdsTdsGdsmCdsAdsGdsGds
36
4974
|
GdsTdsGesAksGesmCksmCe
|
|
1207214
1787
1802
7175
7190
TksGksAdsTdsGdsAdsTdsGdsmCdsmC
26
4975
|
dsTdsTesGksmCesAksGe
|
|
1207222
1893
1908
7281
7296
TksmCksmCdsmCdsTdsGdsAdsGdsmCd
35
3606
|
sAdsAdsTesmCksAesGksGe
|
|
1207230
1901
1916
7289
7304
AksAksGdsAdsTdsGdsAdsGdsTdsmCd
7
2691
|
s
mCdsmCesTksGesAksGe
|
|
1207238
1916
1931
7304
7319
AksTksmCdsAdsmCdsmCdsAdsAdsGds
32
4966
|
GdsAdsGesGksGesAksAe
|
|
1207246
1924
1939
7312
7327
AksmCksTdsGdsmCdsGdsGdsAdsAdsT
17
3607
|
ds
mCdsAesmCksmCesAksAe
|
|
1207254
1932
1947
7320
7335
AksmCksTdsmCdsTdsmCdsTdsmCdsAd
18
4976
|
s
mCdsTdsGesmCksGesGksAe
|
|
1207262
2002
2017
7390
7405
TksGksmCdsGdsmCdsmCdsAdsTdsmCd
37
3305
|
s
mCdsTdsGesGksmCesGksmCe
|
|
1207270
2010
2025
7398
7413
TksGksAdsGdsTdsTdsmCdsmCdsTdsG
21
3913
|
ds
mCdsGesmCksTesAksTe
|
|
1207278
2019
2034
7407
7422
GksmCksAdsmCdsTdsTdsTdsAdsTdsT
7
3000
|
dsGdsAesGksTesTksmCe
|
|
1207286
141
156
508
523
TksGksmCdsTdsmCdsTdsTdsmCdsAds
27
4950
|
GdsCdsTksTesTksGesTk
|
|
1207294
149
164
516
531
mCksGksAdsCdsTdsGdsTdsGdsTdsGd
21
3419
|
s
mCdsTksmCesTksTesmCk
|
|
1207302
157
172
N/A
N/A
AksGksTdsGdsAdsGdsAdsAdsCdsGds
38
2502
|
AdsmCksTesGksTesGk
|
|
1207310
214
229
3565
3580
TksTksTdsGdsTdsGdsGdsTdsAdsmCd
26
2887
|
sAdsGksmCesTksGesmCk
|
|
1207318
222
237
3573
3588
TksGksGdsGdsTdsAdsmCdsAdsTdsTd
30
3421
|
sTdsGksTesGksGesTk
|
|
1207326
1782
1797
7170
7185
AksTksGdsmCdsmCdsTdsTdsGdsmCds
34
4947
|
AdsGdsGksGesTksGesAk
|
|
1207334
1790
1805
7178
7193
AksGksmCdsTdsGdsAdsTdsGdsAdsTd
32
3831
|
sGdsCksCesTksTesGk
|
|
1207342
1896
1911
7284
7299
GksAksGdsTdsmCdsmCdsmCdsTdsGds
33
3834
|
AdsGdsTksAesAksTesTk
|
|
1207350
1904
1919
7292
7307
GksGksAdsAdsAdsGdsAdsTdsGdsAds
54
2921
|
GdsTksmCesmCksmCesTk
|
|
1207358
1919
1934
7307
7322
GksGksAdsAdsTdsCdsAdsCdsCdsAds
8
3226
|
AdsGksGesAksGesGk
|
|
1207366
1927
1942
7315
7330
CksTksCdsAdsCdsTdsGdsCdsGdsGds
13
3835
|
AdsAksTesmCksAesmCk
|
|
1207374
1935
1950
7323
7338
GksmCksmCdsAdsmCdsTdsmCdsTdsmC
31
4967
|
dsTdsmCdsAksmCesTksGesmCk
|
|
1207382
2005
2020
7393
7408
TksmCksmCdsTdsGdsmCdsGdsmCdsmC
27
3533
|
dsAdsTdsmCksmCesTksGesGk
|
|
1207390
2013
2028
7401
7416
TksAksTdsTdsGdsAdsGdsTdsTdsmCd
38
2617
|
s
mCdsTksGesmCksGesmCk
|
|
1207398
2022
2037
7410
7425
AksAksAdsGdsmCdsAdsmCdsTdsTdsT
24
3229
|
dsAdsTksTesGksAesGk
|
|
1207406
144
159
511
526
GksTksGksTdsGdsCdsTdsCdsTdsTds
6
4951
|
CdsAdsGksCksTksTe
|
|
1207414
152
167
N/A
N/A
GksAksAksmCdsGdsAdsCdsTdsGdsT
19
3647
|
dsGdsTdsGksmCksTksmCe
|
|
1207422
209
224
3560
3575
GksGksTksAdsmCdsAdsGdsmCdsTdsG
44
4952
|
ds
mCdsmCdsGksGksTksGe
|
|
1207430
217
232
3568
3583
AksCksAksTdsTdsTdsGdsTdsGdsGds
12
3114
|
TdsAdsmCksAksGksmCe
|
|
1207438
1777
1792
7165
7180
TksTksGksmCdsAdsGdsGdsGdsTdsGd
40
3375
|
sAdsGdsTksTksGksGe
|
|
1207446
1785
1800
7173
7188
AksTksGksAdsTdsGdsmCdsmCdsTdsT
16
3603
|
dsGdsTdsAksGksGksGe
|
|
1207454
1793
1808
7181
7196
mCksmCksmCksAdsGdsmCdsTdsGdsAd
11
4953
|
sTdsGdsAdsTksGksmCksmCe
|
|
1207462
1899
1914
7287
7302
GksAksTksGdsAdsGdsTdsmCdsmCdsm
15
2537
|
CdsTdsGdsAksGksmCksAe
|
|
1207470
1914
1929
7302
7317
CksAksCksCdsAdsAdsGdsGdsAdsGds
40
4954
|
GdsGdsAksAksAksGe
|
|
1207478
1922
1937
7310
7325
TksGksmCksGdsGdsAdsAdsTdsmCdsAds
16
3455
|
mCdsCdsAksAksGksGe
|
|
1207486
1930
1945
7318
7333
TksmCksTksmCdsTdsmCdsAdsmCdsTds
22
4955
|
mGdsCdsGdsGksAksAksTe
|
|
1207494
2000
2015
7388
7403
mCksGksmCksmCdsAdsTdsmCdsmCds
39
3228
|
TdsGdsGdsmCdsGksTksGksGe
|
|
1207502
2008
2023
7396
7411
AksGksTksTdsmCdsmCdsTdsGdsmCds
22
3761
|
GdsmCdsmCdsAksTksmCksmCe
|
|
1207518
2031
2046
7419
7434
AksGksmCksAdsTdsTdsTdsTdsmCds
13
3458
|
AAdsdsAdsGksmCksAksmCe
|
|
TABLE 83
|
|
Reduction of FXII RNA (Huh7, electroporation, 4000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO: 1
NO: 1
NO: 2
NO: 2
|
Compound
Start
Stop
Start
Stop
Sequence and Chemistry
FXII
SEQ
|
Number
Site
Site
Site
Site
notation (5′ to 3′)
(% UTC)
ID NO
|
|
1129531
2009
2024
7397
7412
GksAksGksTdsTdsmCdsmCdsTdsGdsm
8
3837
|
CdsGdsCdsCdsAksTksCk
|
|
1206488
1930
1945
7318
7333
TksmCksTksmCdsTdsmCdsAdsmCdsTd
10
4955
|
sGdsmCdsGdsGdsAksAksTk
|
|
1206497
211
226
3562
3577
GksTksGksGdsTdsAdsmCdsAdsGdsmC
7
4973
|
dsTdsGdsmCdsmCksGksGk
|
|
1206503
1792
1807
7180
7195
mCksmCksAksGdsmCdsTdsGdsAdsTds
35
4970
|
GdsAdsTdsGdsCksCksTk
|
|
1206898
141
156
508
523
TksGesCksTdsCdsTdsTdsCdsAdsGds
18
4950
|
mCdsTdsTesTesGksTk
|
|
1206906
149
164
516
531
mCksGesAksmCdsTdsGdsTdsGdsTdsG
5
3419
|
ds
mCdsTdsmCesTesTesGksTk
|
|
1206914
157
172
N/A
N/A
AksGesTksGdsAdsGdsAdsAdsCdsGds
14
2502
|
AdsmCdsTesGesTksGk
|
|
1206922
214
229
3565
3580
TksTesTksGdsTdsGdsGdsTdsAdsmCd
22
2887
|
sAdsGdsmCesTesGksmCk
|
|
1206930
222
237
3573
3588
TksGesGksGdsTdsAdsmCdsAdsTdsTd
11
3421
|
sTdsGdsTesGesGksTk
|
|
1206938
1782
1797
7170
7185
AksTesGksmCdsmCdsTdsTdsGdsmCds
27
4947
|
AdsGdsGdsGesTesGksAk
|
|
1206946
1790
1805
7178
7193
AksGesmCksTdsGdsAdsTdsGdsAdsTd
32
3831
|
sGdsmCdsmCesTesTksGk
|
|
1206954
1896
1911
7284
7299
GksAesGksTdsmCdsmCdsmCdsTdsGds
16
3834
|
AdsGdsmCdsAesAesTksmCk
|
|
1206962
1904
1919
7292
7307
GksGesAksAdsAdsGdsAdsTdsGdsAds
16
2921
|
GdsTdsmCesmCesmCksTk
|
|
1206970
1919
1934
7307
7322
GksGesAksAdsTdsCdsAdsCdsCdsAds
7
3226
|
AdsGdsGesAesGksGk
|
|
1206978
1927
1942
7315
7330
mCksTesmCksAdsmCdsTdsGdsmCdsGd
7
3835
|
sGdsAdsAdsTesCesAksCk
|
|
1206986
1935
1950
7323
7338
GksmCesmCksAdsmCdsTdsmCdsTdsmC
8
4967
|
dsTdsmCdsAdsmCesTesGksmCk
|
|
1206994
2005
2020
7393
7408
TksmCesmCksTdsGdsmCdsGdsmCdsmC
10
3533
|
dsAdsTdsCdsCesTesGksGk
|
|
1207002
2013
2028
7401
7416
TksAesTksTdsGdsAdsGdsTdsTdsmCd
19
2617
|
s
mCdsTdsGesmCesGksmCk
|
|
1207010
2022
2037
7410
7425
AksAesAksGdsmCdsAdsmCdsTdsTdsT
3
3229
|
dsAdsTdsTesGesAksGk
|
|
1207029
144
159
511
526
GksTksGdsTdsGdsmCdsTdsmCdsTdsT
11
4951
|
d
mCdsAesGesmCesTksTk
|
|
1207037
152
167
N/A
N/A
GksAksAdsmCdsGdsAdsmCdsTdsGdsT
14
3647
|
dsGdsTesGesmCesTksmCk
|
|
1207045
209
224
3560
3575
GksGksTdsAdsmCdsAdsGdsmCdsTdsG
5
4952
|
ds
mCdsTesGesGesTksGk
|
|
1207053
217
232
3568
3583
AksCksAdsTdsTdsTdsGdsTdsGdsGds
12
3114
|
TdsAesmCesAesGksmCk
|
|
1207061
1777
1792
7165
7180
TksTksGdsmCdsAdsGdsGdsGdsTdsGd
20
3375
|
sAdsGesmCesmCesGksGk
|
|
1207082
1785
1800
7173
7188
AksTksGdsAdsTdsGdsmCdsmCdsTdsT
15
3603
|
dsGdsCesAesGesGksGk
|
|
1207090
1793
1808
7181
7196
mCksmCksmCdsAdsGdsmCdsTdsGdsAd
52
4953
|
sTdsGdsAesTesGesmCksmCk
|
|
1207098
1899
1914
7287
7302
GksAksTdsGdsAdsGdsTdsmCdsmCdsm
12
2537
|
CdsTdsGesAesGesmCksAk
|
|
1207106
1914
1929
7302
7317
mCksAksmCdsmCdsAdsAdsGdsGdsAds
13
4954
|
GdsGdsGesAesAesAksGk
|
|
1207114
1922
1937
7310
7325
TksGksCdsGdsGdsAdsAdsTdsCdsAds
11
3455
|
mCdsmCesAesAesGksGk
|
|
1207127
1930
1945
7318
7333
TksmCksTdsmCdsTdsmCdsAdsmCdsTd
12
4955
|
sGdsmCdsGesGesAesAksTk
|
|
1207142
2000
2015
7388
7403
mCksGksmCdsmCdsAdsTdsmCdsmCdsT
24
3228
|
dsGdsGdsmCesGesmCesGksGk
|
|
1207151
2008
2023
7396
7411
AksGksTdsTdsmCdsmCdsTdsGdsmCds
19
3761
|
GdsmCdsmCesAesTesmCksmCk
|
|
1207159
2017
2032
7405
7420
AksmCksTdsTdsTdsAdsTdsTdsGdsAd
4
2848
|
sGdsTesTesCesCksTk
|
|
1207167
2031
2046
7419
7434
AksGksCdsAdsTdsTdsTdsTdsCdsAds
20
3458
|
AdsAesGesCesAksCk
|
|
1207175
147
162
514
529
AksmCksTdsGdsTdsGdsTdsGdsmCdsT
13
3267
|
ds
mCdsTesTksmCesAksGe
|
|
1207183
155
170
N/A
N/A
TksGksAdsGdsAdsAdsCdsGdsAdsCds
26
3875
|
TdsGesTksGesTksGe
|
|
1207191
212
227
3563
3578
TksGksTdsGdsGdsTdsAdsCdsAdsGds
11
2734
|
CdsTesGksmCesmCksGe
|
|
1207199
220
235
3571
3586
GksGksTdsAdsmCdsAdsTdsTdsTdsGd
14
3269
|
sTdsGesGksTesAksmCe
|
|
1207207
1780
1795
7168
7183
GksmCksmCdsTdsTdsGdsmCdsAdsGds
22
3451
|
GdsGdsTesGksAesGksCe
|
|
1207215
1788
1803
7176
7191
mCksTksGdsAdsTdsGdsAdsTdsGdsmC
12
3679
|
ds
mCdsTesTksGesmCksAe
|
|
1207223
1894
1909
7282
7297
GksTksmCdsmCdsmCdsTdsGdsAdsGds
27
3682
|
mCdsAdsAesTksmCesAksGe
|
|
1207231
1902
1917
7290
7305
AksAksAdsGdsAdsTdsGdsAdsGdsTds
19
2768
|
CdsmCesmCksTesGksAe
|
|
1207239
1917
1932
7305
7320
AksAksTdsCdsAdsCdsCdsAdsAdsGds
14
4956
|
GdsAesGksGesGksAe
|
|
1207247
1925
1940
7313
7328
mCksAksmCdsTdsGdsmCdsGdsGdsAds
11
3683
|
AdsTdsmCesAksmCesmCksAe
|
|
1207255
1933
1948
7321
7336
mCksAksmCdsTdsmCdsTdsmCdsTdsmC
N.D.
4957
|
dsAdsmCdsTesGksmCesGksGe
|
|
1207263
2003
2018
7391
7406
mCksTksGdsmCdsGdsmCdsmCdsAdsTd
30
3381
|
s
mCdsmCdsTesGksGesmCksGe
|
|
1207271
2011
2026
7399
7414
TksTksGdsAdsGdsTdsTdsCdsCdsTds
9
2463
|
GdsmCesGksmCesmCksAe
|
|
1207279
2020
2035
7408
7423
AksGksmCdsAdsmCdsTdsTdsTdsAdsT
6
3076
|
dsTdsGesAksGesTksTe
|
|
1207287
142
157
509
524
GksTksGdsmCdsTdsmCdsTdsTdsmCds
15
4958
|
AdsGdsmCksTesTksTesGk
|
|
1207295
150
165
N/A
N/A
AksmCksGdsAdsmCdsTdsGdsTdsGdsT
13
3495
|
dsGdsmCksTesmCksTesTk
|
|
1207303
158
173
N/A
N/A
mCksAksGdsTdsGdsAdsGdsAdsAdsmC
31
2579
|
dsGdsAksmCesTksGesTk
|
|
1207311
215
230
3566
3581
AksTksTdsTdsGdsTdsGdsGdsTdsAds
30
2963
|
mCdsAksGesCksTesGk
|
|
1207319
223
238
3574
3589
GksTksGdsGdsGdsTdsAdsCdsAdsTds
19
3497
|
TdsTksGesTksGesGk
|
|
1207327
1783
1798
7171
7186
GksAksTdsGdsmCdsmCdsTdsTdsGdsm
66
4959
|
CdsAdsGksGesGksTesGk
|
|
1207335
1791
1806
7179
7194
mCksAksGdsmCdsTdsGdsAdsTdsGdsA
24
4960
|
dsTdsGksCesCksTesTk
|
|
1207343
1897
1912
7285
7300
TksGksAdsGdsTdsCdsCdsCdsTdsGds
20
3910
|
AdsGksmCesAksAesTk
|
|
1207351
1905
1920
7293
7308
GksGksGdsAdsAdsAdsGdsAdsTdsGds
10
2997
|
AdsGksTesmCksmCesmCk
|
|
1207359
1920
1935
7308
7323
mCksGksGdsAdsAdsTdsmCdsAdsmCds
18
3303
|
mCdsAdsAksGesGksAesGk
|
|
1207367
1928
1943
7316
7331
TksmCksTdsmCdsAdsmCdsTdsGdsmC
4
3911
|
dsGdsGdsAksAesTksCesAk
|
|
1207375
1936
1951
7324
7339
AksGksmCdsmCdsAdsmCdsTdsmCdsTd
10
4961
|
s
mCdsTdsmCksAesmCksTesGk
|
|
1207383
2006
2021
7394
7409
TksTksTdsCdsTdsGdsCdsGdsCdsCds
20
3609
|
AdsTksCesCksTesGk
|
|
1207391
2014
2029
7402
7417
TksTksAdsTdsTdsGdsAdsGdsTdsTds
28
2694
|
CdsmCksTesGksmCesGk
|
|
1207399
2023
2038
7411
7426
mCksAksAdsAdsGdsmCdsAdsmCdsTds
13
3306
|
TdsTdsAksTesTksGesAk
|
|
1207407
145
160
512
527
TksGksTksGdsTdsGdsCdsTdsCdsTds
6
3112
|
TdsTdsAksGksmCksTe
|
|
1207415
153
168
N/A
N/A
AksGksAksAdsmCdsGdsAdsmCdsTds
19
3723
|
GdsTdsGdsTksGksmCksTe
|
|
1207423
210
225
3561
3576
TksGksGksTdsAds™CdsAdsGdsmCdsT
23
2657
|
dsGdsmCdsmCksGksGksTe
|
|
1207431
218
233
3569
3584
TksAksmCksAdsTdsTdsTdsGdsTdsGd
18
2426
|
sGdsTdsAksmCksAksGe
|
|
1207439
1778
1793
7166
7181
mCksTksTksGdsmCdsAdsGdsGdsGdsT
26
4962
|
dsGdsAdsGksmCksmCksGe
|
|
1207447
1786
1801
7174
7189
GksAksTksGdsAdsTdsGdsmCdsmCdsT
7
4963
|
dsTdsGdsmCksAksGksGe
|
|
1207455
1892
1907
7280
7295
mCksmCksmCksTdsGdsAdsGdsmCdsAd
18
3530
|
sAdsTdsmCdsAksGksGksAe
|
|
1207463
1900
1915
7288
7303
AksGksAksTdsGdsAdsGdsTdsmCdsmC
14
2614
|
ds
mCdsTdsGksAksGksmCe
|
|
1207471
1915
1930
7303
7318
TksmCksAksmCdsmCdsAdsAdsGdsGds
24
4948
|
AdsGdsGdsGksAksAksAe
|
|
1207479
1923
1938
7311
7326
mCksTksGksmCdsGdsGdsAdsAdsTdsm
11
3531
|
CdsAdsmCdsmCksAksAksGe
|
|
1207487
1931
1946
7319
7334
mCksTksmCksTdsmCdsTdsmCdsAdsmC
6
2461
|
dsTdsGdsmCdsGksGksAksAe
|
|
1207495
2001
2016
7389
7404
GksmCksGksmCdsmCdsAdsTdsmCdsmC
116
4964
|
dsTdsGdsGdsmCksGksmCksGe
|
|
1207503
2009
2024
7397
7412
GksAksGksTdsTdsmCdsmCdsTdsGdsm
9
3837
|
CdsGdsmCdsmCksAksTksmCe
|
|
1207511
2018
2033
7406
7421
mCksAksmCksTdsTdsTdsAdsTdsTdsG
11
2924
|
dsAdsGdsTksTksmCksmCe
|
|
1207519
2033
2048
7421
7436
TksmCksAksGdsmCdsAdsTdsTdsTdsT
20
3534
|
ds
mCdsAdsAksAksGksmCe
|
|
TABLE 84
|
|
Reduction of FXII RNA (Huh7, electroporation, 4000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
|
Compound
Start
Stop
Start
Stop
Sequence and Chemistry notation
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(% UTC)
NO
|
|
1129491
1927
1942
7315
7330
mCksTksmCksAdsmCdsTdsGdsmCdsGd
14
3835
|
sGdsAdsAdsTdsmCksAksmCk
|
|
1129531
2009
2024
7397
7412
GksAksGksTdsTdsmCdsmCdsTdsGdsm
7
3837
|
CdsGdsmCdsmCdsAksTksmCk
|
|
1206489
1932
1947
7320
7335
AksmCksTksmCdsTdsmCdsTdsmCdsAd
5
4976
|
s
mCdsTdsGdsmCdsGksGksAk
|
|
1206498
1778
1793
7166
7181
mCksTksTksGdsmCdsAdsGdsGdsGdsT
24
4962
|
dsGdsAdsGdsmCksmCksGk
|
|
1206504
1793
1808
7181
7196
mCksmCksmCksAdsGdsmCdsTdsGdsAd
29
4953
|
sTdsGdsAdsTdsGksmCksmCk
|
|
1206899
142
157
509
524
GksTesGksmCdsTdsmCdsTdsTdsmCds
8
4958
|
AdsGdsmCdsTesTesTksGk
|
|
1206907
150
165
N/A
N/A
AksmCesGksAdsmCdsTdsGdsTdsGdsT
20
3495
|
dsGdsmCdsTesmCesTksTk
|
|
1206915
158
173
N/A
N/A
mCksAesGksTdsGdsAdsGdsAdsAdsmC
25
2579
|
dsGdsAdsCesTesGksTk
|
|
1206923
215
230
3566
3581
AksTesTksTdsGdsTdsGdsGdsTdsAds
23
2963
|
mCdsAdsGesmCesTksGk
|
|
1206931
223
238
3574
3589
GksTesGksGdsGdsTdsAdsmCdsAdsTd
23
3497
|
sTdsTdsGesTesGksGk
|
|
1206939
1783
1798
7171
7186
GksAesTksGdsmCdsmCdsTdsTdsGdsm
32
4959
|
CdsAdsGdsGesGesTksGk
|
|
1206947
1791
1806
7179
7194
mCksAesGksmCdsTdsGdsAdsTdsGdsA
41
4960
|
dsTdsGdsmCesmCesTksTk
|
|
1206955
1897
1912
7285
7300
TksGesAksGdsTdsmCdsmCdsmCdsTds
25
3910
|
GdsAdsGdsCesAesAksTk
|
|
1206963
1905
1920
7293
7308
GksGesGksAdsAdsAdsGdsAdsTdsGds
43
2997
|
AdsGdsTesmCesmCksmCk
|
|
1206971
1920
1935
7308
7323
mCksGesGksAdsAdsTdsmCdsAdsmCds
7
3303
|
mCdsAdsAdsGesGesAksGk
|
|
1206979
1928
1943
7316
7331
TksCesTksCdsAdsCdsTdsGdsCdsGds
15
3911
|
GdsAdsAesTesmCksAk
|
|
1206987
1936
1951
7324
7339
AksGesmCksmCdsAdsmCdsTdsmCdsTd
8
4961
|
s
mCdsTdsmCdsAesmCesTksGk
|
|
1206995
2006
2021
7394
7409
TksTesmCksmCdsTdsGdsmCdsGdsmCd
30
3609
|
s
mCdsAdsTdsmCesmCesTksGk
|
|
1207003
2014
2029
7402
7417
TksTesAksTdsTdsGdsAdsGdsTdsTds
16
2694
|
mCdsmCdsTesGesmCksGk
|
|
1207011
2023
2038
7411
7426
mCksAesAksAdsGdsmCdsAdsmCdsTds
25
3306
|
TdsTdsAdsTesTesGksAk
|
|
1207030
145
160
512
527
TksGksTdsGdsTdsGdsmCdsTdsmCdsT
7
3112
|
dsTdsCesAesGesCksTk
|
|
1207038
153
168
N/A
N/A
AksGksAdsAdsmCdsGdsAdsCdsTdsGd
15
3723
|
sTdsGesTesGesmCksTk
|
|
1207046
210
225
3561
3576
TksGksGdsTdsAdsmCdsAdsGdsmCdsT
18
2657
|
dsGdsmCesmCesGesGksTk
|
|
1207054
218
233
3569
3584
TksAksCdsAdsTdsTdsTdsGdsTdsGds
27
2426
|
GdsTesAesmCesAksGk
|
|
1207062
1778
1793
7166
7181
mCksTksTdsGdsmCdsAdsGdsGdsGdsT
31
4962
|
dsGdsAesGesmCesmCksGk
|
|
1207083
1786
1801
7174
7189
GksAksTdsGdsAdsTdsGdsmCdsmCdsT
18
4963
|
dsTdsGesmCesAesGksGk
|
|
1207091
1892
1907
7280
7295
mCksmCksmCdsTdsGdsAdsGdsmCdsAd
48
3530
|
sAdsTdsmCesAesGesGksAk
|
|
1207099
1900
1915
7288
7303
AksGksAdsTdsGdsAdsGdsTdsmCdsmC
18
2614
|
ds
mCdsTesGesAesGksmCk
|
|
1207107
1915
1930
7303
7318
TksmCksAdsmCdsmCdsAdsAdsGdsGds
20
4948
|
AdsGdsGesGesAesAksAk
|
|
1207115
1923
1938
7311
7326
CksTksGdsCdsGdsGdsAdsAdsTdsmCd
10
3531
|
sAdsCesCesAesAksGk
|
|
1207129
1931
1946
7319
7334
mCksTksmCdsTdsmCdsTdsmCdsAdsmC
8
2461
|
dsTdsGdsmCesGesGesAksAk
|
|
1207144
2001
2016
7389
7404
GksmCksGdsmCdsmCdsAdsTdsmCdsmC
74
4964
|
dsTdsGdsGesmCesGesmCksGk
|
|
1207152
2009
2024
7397
7412
GksAksGdsTdsTdsmCdsmCdsTdsGdsm
16
3837
|
CdsGdsmCesmCesAesTksmCk
|
|
1207160
2018
2033
7406
7421
mCksAksmCdsTdsTdsTdsAdsTdsTdsGd
8
2924
|
sAdsGesTesTesmCksmCk
|
|
1207168
2033
2048
7421
7436
TksmCksAdsGdsmCdsAdsTdsTdsTdsT
29
3534
|
ds
mCdsAesAesAesGksCk
|
|
1207176
148
163
515
530
GksAksmCdsTdsGdsTdsGdsTdsGdsmC
16
3343
|
dsTdsmCesTksTesmCksAe
|
|
1207184
156
171
N/A
N/A
GksTksGdsAdsGdsAdsAdsmCdsGdsAd
18
2424
|
s
mCdsTesGksTesGksTe
|
|
1207192
213
228
3564
3579
TksTksGdsTdsGdsGdsTdsAdsGdsAds
25
2811
|
GdsmCesTksGesmCksmCe
|
|
1207200
221
236
3572
3587
GksGksGdsTdsAdsmCdsAdsTdsTdsTd
51
3345
|
sGdsTesGksGesTksAe
|
|
1207208
1781
1796
7169
7184
TksGksmCdsmCdsTdsTdsGdsmCdsAds
45
3527
|
GdsGdsGesTksGesAksGe
|
|
1207216
1789
1804
7177
7192
GksmCksTdsGdsAdsTdsGdsAdsTdsGd
17
3755
|
s
mCdsmCesTksTesGksmCe
|
|
1207224
1895
1910
7283
7298
AksGksTdsCdsCdsCdsTdsGdsAdsGds
18
3758
|
CdsAesAksTesmCksAe
|
|
1207232
1903
1918
7291
7306
GksAksAdsAdsGdsAdsTdsGdsAdsGds
13
2845
|
TdsCesmCksmCesTksGe
|
|
1207240
1918
1933
7306
7321
GksAksAdsTdsCdsAdsCdsCdsAdsAds
18
4965
|
GdsGesAksGesGksGe
|
|
1207248
1926
1941
7314
7329
TksCksAdsCdsTdsGdsCdsGdsGdsAds
6
3759
|
AdsTesmCksAesmCksmCe
|
|
1207256
1934
1949
7322
7337
mCksmCksAdsmCdsTdsmCdsTdsmCdsT
19
4949
|
ds
mCdsAdsmCesTksGesmCksGe
|
|
1207264
2004
2019
7392
7407
mCksmCksTdsGdsmCdsGdsmCdsmCdsA
18
3457
|
dsTdsmCdsmCesTksGesGksCe
|
|
1207272
2012
2027
7400
7415
AksTksTdsGdsAdsGdsTdsTdsCdsmCd
22
2540
|
sTdsGesmCksGesmCksmCe
|
|
1207280
2021
2036
7409
7424
AksAksGdsCdsAdsCdsTdsTdsTdsAds
9
3147
|
TdsTesGksAesGksTe
|
|
1207288
143
158
510
525
TksGksTdsGdsCdsTdsCdsTdsTdsCds
12
3037
|
AdsGksmCesTksTesTk
|
|
1207296
151
166
N/A
N/A
AksAksCdsGdsAdsCdsTdsGdsTdsGds
33
3571
|
TdsGksmCesTksmCesTk
|
|
1207304
208
223
3559
3574
GksTksAdsmCdsAdsGdsmCdsTdsGdsm
21
4968
|
CdsmCdsGksGesTksGesGk
|
|
1207312
216
231
3567
3582
mCksAksTdsTdsTdsGdsTdsGdsGdsTd
6
3039
|
sAdsmCksAesGksmCesTk
|
|
1207320
1776
1791
7164
7179
TksGksmCdsAdsGdsGdsGdsTdsGdsAd
58
3299
|
sGdsmCksmCesGksGesmCk
|
|
1207328
1784
1799
7172
7187
TksGksAdsTdsGdsmCdsmCdsTdsTdsG
42
4969
|
ds
mCdsAksGesGksGesTk
|
|
1207336
1792
1807
7180
7195
CksCksAdsGdsCdsTdsGdsAdsTdsGds
25
4970
|
AdsTksGesCksCesTk
|
|
1207344
1898
1913
7286
7301
AksTksGdsAdsGdsTdsCdsCdsCdsTds
30
2460
|
GdsAksGesmCksAesAk
|
|
1207352
1906
1921
7294
7309
AksGksGdsGdsAdsAdsAdsGdsAdsTds
18
3073
|
GdsAksGesTksmCesmCk
|
|
1207360
1921
1936
7309
7324
GksmCksGdsGdsAdsAdsTdsmCdsAdsm
6
3379
|
CdsmCdsAksAesGksGesAk
|
|
1207368
1929
1944
7317
7332
mCksTksmCdsTdsmCdsAdsmCdsTdsGd
16
4971
|
s
mCdsGdsGksAesAksTesmCk
|
|
1207376
1937
1952
7325
7340
mCksAksGdsmCdsmCdsAdsmCdsTdsmC
11
4972
|
dsTdsCdsTksmCesAksCesTk
|
|
1207384
2007
2022
7395
7410
GksTksTdsmCdsmCdsTdsGdsmCdsGds
15
3685
|
mCdsmCdsAksTesmCksmCesTk
|
|
1207392
2016
2031
7404
7419
CksTksTdsTdsAdsTdsTdsGdsAdsGds
11
2771
|
TdsTksmCesmCksTesGk
|
|
1207400
2024
2039
7412
7427
TksCksAdsAdsAdsGdsCdsAdsCdsTds
21
3382
|
TdsTksAesTksTesGk
|
|
1207408
146
161
513
528
mCksTksGksTdsGdsTdsGdsmCdsTdsm
20
3190
|
CdsTdsTdsmCksAksGksmCe
|
|
1207416
154
169
N/A
N/A
GksAksGksAdsAdsmCdsGdsAdsmCdsT
24
3799
|
dsGdsTdsGksTksGksmCe
|
|
1207424
211
226
3562
3577
GksTksGksGdsTdsAdsmCdsAdsGdsmC
41
4973
|
dsTdsGdsCksCksGksGe
|
|
1207432
219
234
3570
3585
GksTksAksCdsAdsTdsTdsTdsGdsTds
50
3192
|
GdsGdsTksAksmCksAe
|
|
1207440
1779
1794
7167
7182
mCksmCksTksTdsGdsmCdsAdsGdsGds
34
4974
|
GdsTdsGdsAksGksmCksmCe
|
|
1207448
1787
1802
7175
7190
TksGksAksTdsGdsAdsTdsGdsmCdsmC
25
4975
|
dsTdsTdsGksmCksAksGe
|
|
1207456
1893
1908
7281
7296
TksCksCksCdsTdsGdsAdsGdsCdsAds
21
3606
|
AdsTdsCksAksGksGe
|
|
1207464
1901
1916
7289
7304
AksAksGksAdsTdsGdsAdsGdsTdsmCd
22
2691
|
s
mCdsCdsTksGksAksGe
|
|
1207472
1916
1931
7304
7319
AksTksmCksAdsmCdsmCdsAdsAdsGds
10
4966
|
GdsAdsGdsGksGksAksAe
|
|
1207480
1924
1939
7312
7327
AksCksTksGdsCdsGdsGdsAdsAdsTds
2
3607
|
CdsAdsmCksmCksAksAe
|
|
1207488
1932
1947
7320
7335
AksmCksTksmCdsTdsmCdsTdsCdsAds
6
4976
|
CdsTdsGdsmCksGksGksAe
|
|
1207496
2002
2017
7390
7405
TksGksCksGdsCdsCdsAdsTdsmCdsmC
73
3305
|
dsTdsGdsGksmCksGksmCe
|
|
1207504
2010
2025
7398
7413
TksGksAksGdsTdsTdsCdsCdsTdsGds
5
3913
|
CdsGdsmCksmCksAksTe
|
|
1207512
2019
2034
7407
7422
GksmCksAksmCdsTdsTdsTdsAdsTdsT
3
3000
|
dsGdsAdsGksTksTksmCe
|
|
TABLE 85
|
|
Reduction of FXII RNA (Huh7, electroporation, 4000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
|
Compound
Start
Stop
Start
Stop
Sequence
FXII
SEQ ID
|
Number
Site
Site
Site
Site
and Chemistry notation (5′ to 3′)
(% UTC)
NO
|
|
1129531
2009
2024
7397
7412
GksAksGksTdsTdsmCdsmCdsTdsGdsm
15
3837
|
CdsGdsmCdsmCdsAksTksmCk
|
|
1129532
2010
2025
7398
7413
TksGksAksGdsTdsTdsmCdsmCdsTdsG
7
3913
|
ds
mCdsGdsCdsCksAksTk
|
|
1206485
1914
1929
7302
7317
mCksAksmCksmCdsAdsAdsGdsGdsAds
27
4954
|
GdsGdsGdsAdsAksAksGk
|
|
1206490
1933
1948
7321
7336
mCksAksmCksTdsmCdsTdsmCdsTdsmC
6
4957
|
dsAdsmCdsTdsGdsmCksGksGk
|
|
1206499
1779
1794
7167
7182
mCksmCksTksTdsGdsmCdsAdsGdsGds
36
4974
|
GdsTdsGdsAdsGksmCksmCk
|
|
1206900
143
158
510
525
TksGesTksGdsCdsTdsCdsTdsTdsCds
24
3037
|
AdsGdsCesTesTksTk
|
|
1206908
151
166
N/A
N/A
AksAesmCksGdsAdsmCdsTdsGdsTdsG
12
3571
|
dsTdsGdsmCesTesmCksTk
|
|
1206916
208
223
3559
3574
GksTesAksmCdsAdsGdsmCdsTdsGdsm
32
4968
|
CdsmCdsGdsGesTesGksGk
|
|
1206924
216
231
3567
3582
TksAesTksTdsTdsGdsTdsGdsGdsTds
26
3039
|
AdsmCdsAesGesmCksTk
|
|
1206932
1776
1791
7164
7179
TksGesmCksAdsGdsGdsGdsTdsGdsAd
40
3299
|
sGdsmCdsmCesGesGksmCk
|
|
1206940
1784
1799
7172
7187
TksGesAksTdsGdsmCdsmCdsTdsTdsG
25
4969
|
ds
mCdsAdsGesGesGksTk
|
|
1206948
1792
1807
7180
7195
mCksmCesAksGdsmCdsTdsGdsAdsTds
24
4970
|
GdsAdsTdsGesmCesmCksTk
|
|
1206956
1898
1913
7286
7301
AksTesGksAdsGdsTdsmCdsmCdsmCds
16
2460
|
TdsGdsAdsGesmCesAksAk
|
|
1206964
1906
1921
7294
7309
AksGesGksGdsAdsAdsAdsGdsAdsTds
21
3073
|
GdsAdsGesTesmCksmCk
|
|
1206972
1921
1936
7309
7324
GksmCesGksGdsAdsAdsTdsmCdsAdsm
15
3379
|
CdsmCdsAdsAesGesGksAk
|
|
1206980
1929
1944
7317
7332
mCksTesmCksTdsmCdsAdsmCdsTdsGd
8
4971
|
s
mCdsGdsGdsAesAesTksmCk
|
|
1206988
1937
1952
7325
7340
mCksAesGksmCdsmCdsAdsmCdsTdsmC
15
4972
|
dsTdsmCdsTdsmCesAesmCksTk
|
|
1206996
2007
2022
7395
7410
GksTesTksmCdsmCdsTdsGdsmCdsGds
10
3685
|
mCdsmCdsAdsTesmCesmCksTk
|
|
1207004
2016
2031
7404
7419
mCksTesTksTdsAdsTdsTdsGdsAdsGd
7
2771
|
sTdsTdsmCesmCesTksGk
|
|
1207012
2024
2039
7412
7427
TksmCesAksAdsAdsGdsmCdsAdsmCds
14
3382
|
TdsTdsTdsAesTesTksGk
|
|
1207031
146
161
513
528
mCksTksGdsTdsGdsTdsGdsmCdsTdsm
24
3190
|
CdsTdsTesmCesAesGksmCk
|
|
1207039
154
169
N/A
N/A
GksAksGdsAdsAdsmCdsGdsAdsmCdsT
14
3799
|
dsGdsTesGesTesGksmCk
|
|
1207047
211
226
3562
3577
GksTksGdsGdsTdsAdsmCdsAdsGdsmC
18
4973
|
dsTdsGesmCesmCesGksGk
|
|
1207055
219
234
3570
3585
GksTksAdsmCdsAdsTdsTdsTdsGdsTd
65
3192
|
sGdsGesTesAesmCksAk
|
|
1207063
1779
1794
7167
7182
mCksmCksTdsTdsGdsmCdsAdsGdsGds
32
4974
|
GdsTdsGesAesGesmCksmCk
|
|
1207084
1787
1802
7175
7190
TksGksAdsTdsGdsAdsTdsGdsmCdsmC
15
4975
|
dsTdsTesGesmCesAksGk
|
|
1207092
1893
1908
7281
7296
TksmCksmCdsmCdsTdsGdsAdsGdsmCd
29
3606
|
sAdsAdsTesmCesAesGksGk
|
|
1207100
1901
1916
7289
7304
AksAksGdsAdsTdsGdsAdsGdsTdsmCd
16
2691
|
s
mCdsmCesTesGesAksGk
|
|
1207108
1916
1931
7304
7319
AksTksmCdsAdsmCdsmCdsAdsAdsGds
9
4966
|
GdsAdsGesGesGesAksAk
|
|
1207116
1924
1939
7312
7327
AksmCksTdsGdsmCdsGdsGdsAdsAdsT
10
3607
|
ds
mCdsAEsmCesmCesAksAk
|
|
1207131
1932
1947
7320
7335
AksmCksTdsmCdsTdsmCdsTdsmCdsAd
6
4976
|
s
mCdsTdsGesmCesGesGksAk
|
|
1207145
2002
2017
7390
7405
TksGksmCdsGdsmCdsmCdsAdsTdsmCd
46
3305
|
s
mCdsTdsGesGesmCesGksmCk
|
|
1207153
2010
2025
7398
7413
TksGksAdsGdsTdsTdsmCdsmCdsTdsG
13
3913
|
ds
mCdsGesCesCesAksTk
|
|
1207161
2019
2034
7407
7422
GksmCksAdsmCdsTdsTdsTdsAdsTdsT
17
3000
|
dsGdsAesGesTesTksmCk
|
|
1207169
141
156
508
523
TksGksmCdsTdsmCdsTdsTdsmCdsAds
33
4950
|
GdsmCdsTesTksTesGksTe
|
|
1207177
149
164
516
531
mCksGksAdsmCdsTdsGdsTdsGdsTdsG
17
3419
|
ds
mCdsTesmCksTesTksmCe
|
|
1207185
157
172
N/A
N/A
AksGksTdsGdsAdsGdsAdsAdsmCdsGd
15
2502
|
sAdsmCesTksGesTksGe
|
|
1207193
214
229
3565
3580
TksTksTdsGdsTdsGdsGdsTdsAdsCds
17
2887
|
AdsGesmCksTesGksmCe
|
|
1207201
222
237
3573
3588
TksGksGdsGksTdsAdsCdsAdsTdsTd
25
3421
|
sTdsGesTksGesGksTe
|
|
1207209
1782
1797
7170
7185
AksTksGdsmCdsmCdsTdsTdsGdsmCds
26
4947
|
AdsGdsGesGksTesGksAe
|
|
1207217
1790
1805
7178
7193
AksGksmCdsTdsGdsAdsTdsGdsAdsTd
22
3831
|
sGdsmCesmCksTesTksGe
|
|
1207225
1896
1911
7284
7299
GksAksGdsTdsmCdsmCdsmCdsTdsGds
7
3834
|
AdsGdsCesAksAesTksCe
|
|
1207233
1904
1919
7292
7307
GksGksAdsAdsAdsGdsAdsTdsGdsAds
31
2921
|
GdsTesCksCesCksTe
|
|
1207241
1919
1934
7307
7322
GksGksAdsAdsTdsCdsAdsmCdsCdsAd
21
3226
|
sAdsGesGksAesGksGe
|
|
1207249
1927
1942
7315
7330
mCksTksmCdsAdsmCdsTdsGdsmCdsGd
12
3835
|
sGdsAdsAesTksmCesAksmCe
|
|
1207257
1935
1950
7323
7338
GksmCksmCdsAdsmCdsTdsmCdsTdsmC
25
4967
|
dsTdsmCdsAesmCksTesGksmCe
|
|
1207265
2005
2020
7393
7408
TksCksCdsTdsGdsCdsGdsCdsCdsAds
24
3533
|
TdsmCesmCksTesGksGe
|
|
1207273
2013
2028
7401
7416
TksAksTdsTdsGdsAdsGdsTdsTdsmCd
21
2617
|
s
mCdsTesGksmCesGksmCe
|
|
1207281
2022
2037
7410
7425
AksAksAdsGdsmCdsAdsmCdsTdsTdsT
21
3229
|
dsAdsTesTksGesAksGe
|
|
1207289
144
159
511
526
GksTksGdsTdsGdsmCdsTdsmCdsTdsT
20
4951
|
ds
mCdsAksGesmCksTesTk
|
|
1207297
152
167
N/A
N/A
GksAksAdsmCdsGdsAdsmCdsTdsGdsT
23
3647
|
dsGdsTksGesCksTesCk
|
|
1207305
209
224
3560
3575
GksGksTdsAdsmCdsAdsGdsCdsTdsGd
23
4952
|
sCdsmCksGesGksTesGk
|
|
1207313
217
232
3568
3583
AksCksAdsTdsTdsTdsGdsTdsGdsGds
12
3114
|
TdsAksmCesAksGesmCk
|
|
1207321
1777
1792
7165
7180
TksTksGdsmCdsAdsGdsGdsGdsTdsGd
36
3375
|
sAdsGksmCesmCksGesGk
|
|
1207329
1785
1800
7173
7188
AksTksGdsAdsTdsGdsCdsCdsTdsTds
21
3603
|
GdsmCksAesGksGesGk
|
|
1207337
1793
1808
7181
7196
mCksmCksmCdsAdsGdsmCdsTdsGdsAd
53
4953
|
sTdsGdsAksTesGksmCesmCk
|
|
1207345
1899
1914
7287
7302
GksAksTdsGdsAdsGdsTdsmCdsmCdsm
12
2537
|
CdsTdsGksAesGksCesAk
|
|
1207353
1914
1929
7302
7317
mCksAksmCdsmCdsAdsAdsGdsGdsAds
22
4954
|
GdsGdsGksAesAksAesGk
|
|
1207361
1922
1937
7310
7325
TksGksmCdsGdsGdsAdsAdsTdsmCdsA
11
3455
|
ds
mCdsmCksAesAksGesGk
|
|
1207369
1930
1945
7318
7333
TksCksTdsCdsTdsCdsAdsCdsTdsGds
13
4955
|
mCdsGksGesAksAesTk
|
|
1207377
2000
2015
7388
7403
mCksGksmCdsmCdsAdsTdsmCdsmCdsT
42
3228
|
dsGdsGdsmCksGesmCksGesGk
|
|
1207385
2008
2023
7396
7411
AksGksTdsTdsmCdsmCdsTdsGdsmCds
14
3761
|
GdsmCdsmCksAesTksmCesmCk
|
|
1207393
2017
2032
7405
7420
AksmCksTdsTdsTdsAdsTdsTdsGdsAd
7
2848
|
sGdsTksTesCksCesTk
|
|
1207401
2031
2046
7419
7434
AksGksCdsAdsTdsTdsTdsTdsCdsAds
10
3458
|
AdsAksGesmCksAesmCk
|
|
1207409
147
162
514
529
AksmCksTksGdsTdsGdsTdsGdsmCdsT
21
3267
|
ds
mCdsTdsTksmCksAksGe
|
|
1207417
155
170
N/A
N/A
TksGksAksGdsAdsAdsmCdsGdsAdsmC
35
3875
|
dsTdsGdsTksGksTksGe
|
|
1207425
212
227
3563
3578
TksGksTksGdsGdsTdsAdsmCdsAdsGd
30
2734
|
s
mCdsTdsGksmCksmCksGe
|
|
1207433
220
235
3571
3586
GksGksTksAdsmCdsAdsTdsTdsTdsGd
23
3269
|
sTdsGdsGksTksAksmCe
|
|
1207441
1780
1795
7168
7183
GksmCksmCksTdsTdsGdsmCdsAdsGds
26
3451
|
GdsGdsTdsGksAksGksmCe
|
|
1207449
1788
1803
7176
7191
mCksTksGksAdsTdsGdsAdsTdsGdsmC
25
3679
|
ds
mCdsTdsTksGksmCksAe
|
|
1207457
1894
1909
7282
7297
GksTksmCksmCdsmCdsTdsGdsAdsGds
25
3682
|
mCdsAdsAdsTksmCksAksGe
|
|
1207465
1902
1917
7290
7305
AksAksAksGdsAdsTdsGdsAdsGdsTds
13
2768
|
CdsmCdsmCksTksGksAe
|
|
1207473
1917
1932
7305
7320
AksAksTksCdsAdsCdsCdsAdsAdsGds
23
4956
|
GdsAdsGksGksGksAe
|
|
1207481
1925
1940
7313
7328
mCksAksmCksTdsGdsmCdsGdsGdsAds
4
3683
|
AdsTdsmCdsAksmCksmCksAe
|
|
1207489
1933
1948
7321
7336
mCksAksmCksTdsmCdsTdsmCdsTdsmC
12
4957
|
dsAdsmCdsTdsGksmCksGksGe
|
|
1207497
2003
2018
7391
7406
mCksTksGksmCdsGdsmCdsmCdsAdsTd
45
3381
|
s
mCdsmCdsTdsGksGksmCksGe
|
|
1207505
2011
2026
7399
7414
TksTksGksAdsGdsTdsTdsmCdsmCdsT
7
2463
|
dsGdsmCdsGksmCksmCksAe
|
|
1207513
2020
2035
7408
7423
AksGksCksAdsmCdsTdsTdsTdsAdsT
4
3076
|
dsTdsGdsAksGksTksTe
|
|
TABLE 86
|
|
Reduction of FXII RNA (Huh7, electroporation, 2000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
|
Compound
Start
Stop
Start
Stop
Sequence and Chemistry notation
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(%UTC)
NO
|
|
1128769
153
168
N/A
N/A
AksGksAksAdsmCdsGdsAdsmCdsTdsGds
19
3723
|
TdsGdsTdsGksmCksTk
|
|
1128802
218
233
3569
3584
TksAksmCksAdsTdsTdsTdsGdsTdsGdsGds
108
2426
|
TdsAdsmCksAksGk
|
|
1128870
470
485
4415
4430
TksGksTksGdsGdsAdsAdsAdsAdsAdsmCds
31
3728
|
mCdsGdsGksAksGk
|
|
1129485
1921
1936
7309
7324
GksmCksGksGdsAdsAdsTdsmCdsAdsmCds
20
3379
|
mCdsAdsAdsGksGksAk
|
|
1129491
1927
1942
7315
7330
mCksTksmCksAdsmCdsTdsGdsmCdsGdsGds
17
3835
|
AdsAdsTdsmCksAksmCk
|
|
1129492
1928
1943
7316
7331
TksmCksTksmCdsAdsmCdsTdsGdsmCdsGds
7
3911
|
GdsAdsAdsTksmCksAk
|
|
1129531
2009
2024
7397
7412
GksAksGksTdsTdsmCdsmCdsTdsGdsmCds
17
3837
|
GdsmCdsmCdsAksTksmCk
|
|
1129532
2010
2025
7398
7413
TksGksAksGdsTdsTdsmCdsmCdsTdsGdsm
6
3913
|
CdsGdsmCdsmCksAksTk
|
|
1129538
2017
2032
7405
7420
AksmCksTksTdsTdsAdsTdsTdsGdsAdsGds
7
2848
|
TdsTdsmCksmCksTk
|
|
1129540
2019
2034
7407
7422
GksmCksAksmCdsTdsTdsTdsAdsTdsTdsGds
3
3000
|
AdsGdsTksTksmCk
|
|
1129579
N/A
N/A
208
223
mCksAksmCksAdsTdsAdsGdsGdsmCdsmCds
55
2926
|
TdsmCdsmCdsTksAksGk
|
|
1129714
N/A
N/A
1558
1573
AksGksGksGdsTdsAdsAdsGdsGdsTdsmCds
63
2549
|
mCdsAdsTksmCksTk
|
|
1129740
N/A
N/A
1758
1773
GksAksTksGdsGdsGdsAdsTdsGdsGdsAdsm
69
3010
|
CdsGdsGksAksmCk
|
|
1129760
N/A
N/A
2284
2299
AksmCksmCksAdsTdsmCdsGdsmCdsGdsAds
44
3011
|
AdsTdsAdsAksTksGk
|
|
1129773
N/A
N/A
2450
2465
GksmCksAksTdsTdsGdsAdsTdsGdsAdsAds
28
2475
|
TdsmCdsAksGksmCk
|
|
1129779
N/A
N/A
2470
2485
GksmCksTksAdsTdsAdsGdsTdsGdsmCdsm
27
2936
|
CdsAdsTdsmCksAksGk
|
|
1129802
N/A
N/A
2654
2669
TksmCksmCksTdsAdsTdsTdsGdsAdsAdsTds
33
3160
|
GdsAdsGksTksGk
|
|
1129948
N/A
N/A
4840
4855
AksTksmCksTdsGdsmCdsTdsTdsTdsmCdsm
51
3630
|
CdsGdsmCdsAksmCksTk
|
|
1129949
N/A
N/A
4844
4859
TksmCksTksmCdsAdsTdsmCdsTdsGdsmCds
24
3706
|
TdsTdsTdsmCksmCksGk
|
|
1130060
N/A
N/A
6178
6193
AksAksGksGdsmCdsGdsmCdsAdsAdsmCds
22
3026
|
AdsGdsAdsGksmCksTk
|
|
1130240
N/A
N/A
7098
7113
GksTksmCksGdsGdsAdsAdsAdsmCdsAdsm
47†
3035
|
CdsGdsmCdsAksGksmCk
|
|
1213035
141
156
508
523
TksGksmCksTdsCysTdsTdsmCdsAdsGddsm
40
4950
|
CdsTdsTdsTksGksTk
|
|
1213036
142
157
509
524
GksTksGksmCdsUysmCdsTdsTdsmCdsAds
28
4977
|
GdsmCdsTdsTksTksGk
|
|
1213037
143
158
510
525
TksGksTksGdsCysTdsmCdsTdsTdsmCdsAds
46
3037
|
GdsmCdsTksTksTk
|
|
1213038
144
159
511
526
GksTksGksTdsGysmCdsTdsmCdsTdsTdsm
33
4951
|
CdsAdsGdsmCksTksTk
|
|
1213039
145
160
512
527
TksGksTksGdsUysGdsmCdsTdsmCdsTdsTds
91
4978
|
mCdsAdsGksmCksTk
|
|
1213040
146
161
513
528
mCksTksGksTdsGysTdsGdsmCdsTdsmCds
35
3190
|
TdsTdsmCdsAksGksmCk
|
|
1213041
147
162
514
529
AksmCksTksGdsUysGdsTdsGdsmCdsTdsm
27
4979
|
CdsTdsTdsmCksAksGk
|
|
1213042
148
163
515
530
GksAksmCksTdsGysTdsGdsTdsGdsmCdsTds
50
3343
|
mCdsTdsTksmCksAk
|
|
1213043
149
164
516
531
mCksGksAksmCdsUysGdsTdsGdsTdsGdsm
65
4980
|
CdsTdsmCdsTksTksmCk
|
|
1213044
150
165
N/A
N/A
AksmCksGksAdsCysTdsGdsTdsGdsTdsGds
52
3495
|
mCdsTdsmCksTksTk
|
|
1213045
151
166
N/A
N/A
AksAksmCksGdsAysmCdsTdsGdsTdsGdsTds
47
3571
|
GdsmCdsTksmCksTk
|
|
1213046
152
167
N/A
N/A
GksAksAksmCdsGysAdsmCdsTdsGdsTds
77
3647
|
GdsTdsGdsmCksTksmCk
|
|
1213047
153
168
N/A
N/A
AksGksAksAdsCysGdsAdsmCdsTdsGdsTds
42
3723
|
GdsTdsGksmCksTk
|
|
1213048
154
169
N/A
N/A
GksAksGksAdsAysmCdsGdsAdsmCdsTdsGds
61
3799
|
TdsGdsTksGksmCk
|
|
1213049
155
170
N/A
N/A
TksGksAksGdsAysAdsmCdsGdsAdsmCds
42
3875
|
TdsGdsTdsGksTksGk
|
|
1213050
156
171
N/A
N/A
GksTksGksAdsGysAdsAdsmCdsGdsAdsm
35
2424
|
CdsTdsGdsTksGksTk
|
|
1213051
157
172
N/A
N/A
AksGksTksGdsAysGdsAdsAdsmCdsGdsAds
69
2502
|
mCdsTdsGksTksGk
|
|
1213052
158
173
N/A
N/A
mCksAksGksTdsGysAdsGdsAdsAdsmCds
54
2579
|
GdsAdsmCdsTksGksTk
|
|
1213053
208
223
3559
3574
GksTksAksmCdsAysGdsmCdsTdsGdsmCdsm
33
4968
|
CdsGdsGdsTksGksGk
|
|
1213054
209
224
3560
3575
GksGksTksAdsCysAdsGdsmCdsTdsGdsm
33
4952
|
CdsmCdsGdsGksTksGk
|
|
1213055
210
225
3561
3576
TksGksGksTdsAysmCdsAdsGdsmCdsTdsGds
26
2657
|
mCdsmCdsGksGksTk
|
|
1213056
211
226
3562
3577
GksTksGksGdsUysAdsmCdsAdsGdsmCds
34
4981
|
TdsGdsmCdsmCksGksGk
|
|
1213057
212
227
3563
3578
TksGksTksGdsGysTdsAdsmCdsAdsGdsmCds
78
2734
|
TdsGdsmCksmCksGk
|
|
1213058
213
228
3564
3579
TksTksGksTdsGysGdsTdsAdsmCdsAdsGds
29
2811
|
mCdsTdsGksmCksmCk
|
|
1213059
214
229
3565
3580
TksTksTksGdsUysGdsGdsTdsAdsmCdsAds
44
4982
|
GdsmCdsTksGksmCk
|
|
1213060
215
230
3566
3581
AksTksTksTdsGysTdsGdsGdsTdsAdsmCds
44
2963
|
AdsGdsmCksTksGk
|
|
1213061
216
231
3567
3582
mCksAksTksTdsUysGdsTdsGdsGdsTdsAds
36
4983
|
mCdsAdsGksmCksTk
|
|
1213062
217
232
3568
3583
AksmCksAksTdsUysTdsGdsTdsGdsGdsTds
75
4984
|
AdsmCdsAksGksmCk
|
|
1213063
218
233
3569
3584
TksAksmCksAdsUysTdsTdsGdsTdsGdsGds
55
4985
|
TdsAdsmCksAksGk
|
|
1213064
219
234
3570
3585
GksTksAksmCdsAysTdsTdsTdsGdsTdsGds
38
3192
|
GdsTdsAksmCksAk
|
|
1213065
220
235
3571
3586
GksGksTksAdsCysAdsTdsTdsTdsGdsTds
54
3269
|
GdsGdsTksAksmCk
|
|
1213066
221
236
3572
3587
GksGksGksTdsAysmCdsAdsTdsTdsTdsGds
24
3345
|
TdsGdsGksTksAk
|
|
1213067
222
237
3573
3588
TksGksGksGdsUysAdsmCdsAdsTdsTdsTds
31
4986
|
GdsTdsGksGksTk
|
|
1213068
223
238
3574
3589
GksTksGksGdsGysTdsAdsmCdsAdsTdsTds
63
3497
|
TdsGdsTksGksGk
|
|
1213069
1776
1791
7164
7179
TksGksmCksAdsGysGdsGdsTdsGdsAdsGds
26
3299
|
mCdsmCdsGksGksmCk
|
|
1213070
1777
1792
7165
7180
TksTksGksmCdsAysGdsGdsGdsTdsGdsAds
38
3375
|
GdsmCdsmCksGksGk
|
|
1213071
1778
1793
7166
7181
mCksTksTksGdsCysAdsGdsGdsGdsTdsGds
24
4962
|
AdsGdsmCksmCksGk
|
|
1213072
1779
1794
7167
7182
mCksmCksTksTdsGysmCdsAdsGdsGdsGds
41
4974
|
TdsGdsAdsGksmCksmCk
|
|
1213073
1780
1795
7168
7183
GksmCksmCksTdsUysGdsmCdsAdsGdsGds
25
4987
|
GdsTdsGdsAksGksmCk
|
|
1213074
1781
1796
7169
7184
TksGksmCksmCdsUysTdsGdsmCdsAdsGds
16
4988
|
GdsGdsTdsGksAksGk
|
|
1213075
1782
1797
7170
7185
AksTksGksmCdsCysTdsTdsGdsmCdsAdsGds
26
4947
|
GdsGdsTksGksAk
|
|
1213076
1783
1798
7171
7186
GksAksTksGdsCysmCdsTdsTdsGdsmCdsAds
23
4959
|
GdsGdsGksTksGk
|
|
1213077
1784
1799
7172
7187
AksGksAksAdsGysmCdsmCdsTdsTdsGdsm
17
4969
|
CdsAdsGdsGksGksTk
|
|
1213078
1785
1800
7173
7188
AksTkGksAdsUysGdsmCdsmCdsTdsTdsGds
25
4989
|
mCdsAdsGksGksGk
|
|
1213079
1786
1801
7174
7189
GksAksTksGdsAysTdsGdsmCdsmCdsTdsTds
24
4963
|
GdsmCdsAksGksGk
|
|
1213080
1787
1802
7175
7190
TksGksAksTdsGysAdsTdsGdsmCdsmCdsTds
34
4975
|
TdsGdsmCksAksGk
|
|
1213081
1788
1803
7176
7191
mCksTksGksAdsUysGdsAdsTdsGdsmCdsm
19
4990
|
CdsTdsTdsGksmCksAk
|
|
1213082
1789
1804
7177
7192
GksmCksTksGdsAysTdsGdsAdsTdsGdsmCds
50
3755
|
mCdsTdsTksGksmCk
|
|
1213083
1790
1805
7178
7193
AksGksmCksTdsGysAdsTdsGdsAdsTdsGds
59
3831
|
mCdsmCdsTksTksGk
|
|
1213084
1791
1806
7179
7194
mCksAksGksmCdsUysGdsAdsTdsGdsAds
35
4991
|
TdsGdsmCdsmCksTksTk
|
|
1213085
1792
1807
7180
7195
mCksmCksAksGdsCysTdsGdsAdsTdsGds
31
4970
|
AdsTdsGdsmCksCksTk
|
|
1213086
1793
1808
7181
7196
mCksmCksmCksAdsGysmCdsTdsGdsAdsTds
39
4953
|
GdsAdsTdsGksmCksmCk
|
|
1213087
1892
1907
7280
7295
mCksmCksmCksTdsGysAdsGdsmCdsAdsAds
27
3530
|
TdsmCdsAdsGksGksAk
|
|
1213088
1893
1908
7281
7296
TksmCksmCksmCdsUysGdsAdsGdsmCdsAds
43
4992
|
AdsTdsmCdsAksGksGk
|
|
1213089
1894
1909
7282
7297
GksTksmCksmCdsCysTdsGdsAdsGdsmCds
22
3682
|
AdsAdsTdsmCksAksGk
|
|
1213090
1895
1910
7283
7298
AksGksTksmCdsCysmCdsTdsGdsAdsGdsm
29
3758
|
CdsAdsAdsTksmCksAk
|
|
1213091
1896
1911
7284
7299
GksAksGksTdsCysmCdsmCdsTdsGdsAds
33
3834
|
GdsmCdsAdsAksTksmCk
|
|
1213092
1897
1912
7285
7300
TksGksAksGdsUysmCdsmCdsmCdsTdsGds
38
4993
|
AdsGdsmCdsAksAksTk
|
|
TABLE 87
|
|
Reduction of FXII RNA (Huh7, electroporation, 2000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
|
Compound
Start
Stop
Start
Stop
Sequence and Chemistry notation
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(%UTC)
NO
|
|
1128769
153
168
N/A
N/A
AksGksAksAdsmCdsGdsAdsmCdsTdsGds
63
3723
|
TdsGdsTdsGksmCksTk
|
|
1128802
218
233
3569
3584
TksAksmCksAdsTdsTdsTdsGdsTdsGdsGds
90
2426
|
TdsAdsmCksAksGk
|
|
1128870
470
485
4415
4430
TksGksTksGdsGdsAdsAdsAdsAdsAdsmCds
44
3728
|
mCdsGdsGksAksGk
|
|
1129409
1788
1803
7176
7191
mCksTksGksAdsTdsGdsAdsTdsGdsmCdsm
7
3679
|
CdsTdsTdsGksmCksAk
|
|
1129486
1922
1937
7310
7325
TksGksmCksGdsGdsAdsAdsTdsmCdsAds™
11
3455
|
CdsmCdsAdsAksGksGk
|
|
1129489
1925
1940
7313
7328
mCksAksmCksTdsGdsmCdsGdsGdsAdsAds
21
3683
|
TdsmCdsAdsmCksmCksAk
|
|
1129491
1927
1942
7315
7330
mCksTksmCksAdsCdsTdsGdsmCdsGdsGds
54
3835
|
AdsAdsTdsmCksAksmCk
|
|
1129531
2009
2024
7397
7412
GksAksGksTdsTdsmCdsmCdsTdsGdsmCds
9
3837
|
GdsmCdsmCdsAksTksmCk
|
|
1129532
2010
2025
7398
7413
TksGksAksGdsTdsTdsmCdsmCdsTdsGdsm
16
3913
|
CdsGdsmCdsmCksAksTk
|
|
1129540
2019
2034
7407
7422
GksmCksAksmCdsTdsTdsTdsAdsTdsTdsGds
19
3000
|
AdsGdsTksTksmCk
|
|
1129572
N/A
N/A
186
201
GksGksAksmCdsmCdsmCdsAdsmCdsAdsGds
47
3915
|
GdsTdsmCdsAksTksGk
|
|
1129857
N/A
N/A
3881
3896
GksTksAksAdsTdsGdsAdsGdsGdsCdsGds
48
2787
|
GdsGdsAksGksGk
|
|
1129938
N/A
N/A
4635
4650
AksmCksGksmCdsAdsGdsTdsGdsAdsGdsm
62
2868
|
CdsmCdsAdsmCksmCksmCk
|
|
1129948
N/A
N/A
4840
4855
AksTksmCksTdsGdsmCdsTdsTdsTdsmCdsm
34
3630
|
CdsGdsmCdsAksmCksTk
|
|
1129949
N/A
N/A
4844
4859
TksmCksTksmCdsAdsTdsmCdsTdsGdsmCds
41
3706
|
TdsTdsTdsmCksmCksGk
|
|
1130010
N/A
N/A
5752
5767
AksmCksGksGdsAdsGdsGdsAdsGdsmCdsm
29
3785
|
CdsGdsmCdsGksGksmCk
|
|
1130142
N/A
N/A
6579
6594
mCksGksTksmCdsmCdsTdsAdsGdsTdsTds
32
3177
|
GdsGdsmCdsmCksTksTk
|
|
1130215
N/A
N/A
7015
7030
CksAksmCksmCdsTdsGdsGdsmCdsAdsm
41†
2651
|
CdsGdsmCdsAdsTksmCksGk
|
|
1130251
N/A
N/A
7111
7126
GksGksAksAdsTdsmCdsAdsmCdsmCdsmCds
30†
3873
|
TdsGdsGdsGksTksmCk
|
|
1130252
N/A
N/A
7112
7127
mCksGksGksAdsAdsTdsmCdsAdsmCdsmCds
15†
3949
|
mCdsTdsGdsGksGksTk
|
|
1213093
1898
1913
7286
7301
AksTksGksAdsGysTdsmCdsmCdsmCdsTds
42
2460
|
GdsAdsGdsmCksAksAk
|
|
1213094
1899
1914
7287
7302
GksAksTksGdsAysGdsTdsmCdsmCdsmCds
18
2537
|
TdsGdsAdsGksmCksAk
|
|
1213095
1900
1915
7288
7303
AksGksAksTdsGysAdsGdsTdsmCdsmCdsm
13
2614
|
CdsTdsGdsAksGksmCk
|
|
1213096
1901
1916
7289
7304
AksAkGksAdsUysGdsAdsGdsTdsmCdsm
28
4994
|
CdsmCdsTdsGksAksGk
|
|
1213097
1902
1917
7290
7305
AksAksAksGdsAysTdsGdsAdsGdsTdsmCds
19
2768
|
mCdsmCdsTksGksAk
|
|
1213098
1903
1918
7291
7306
GksAksAksAdsGysAdsTdsGdsAdsGdsTdsm
36
2845
|
CdsmCdsmCksTksGk
|
|
1213099
1904
1919
7292
7307
GksGksAksAdsAysGdsAdsTdsGdsAdsGds
25
2921
|
TdsmCdsmCksmCksTk
|
|
1213100
1905
1920
7293
7308
GksGksGksAdsAysAdsGdsAdsTdsGdsAds
26
2997
|
GdsTdsmCksmCksmCk
|
|
1213101
1906
1921
7294
7309
AksGksGksGdsAysAdsAdsGdsAdsTdsGds
24
3073
|
AdsGdsTksmCksmCk
|
|
1213102
1914
1929
7302
7317
mCksAksmCksmCdsAysAdsGdsGdsAdsGds
34
4954
|
GdsGdsAdsAksAksGk
|
|
1213103
1915
1930
7303
7318
TksmCksAksmCdsCysAdsAdsGdsGdsAdsGds
41
4948
|
GdsGdsAksAksAk
|
|
1213104
1916
1931
7304
7319
AksTksmCksAdsCysmCdsAdsAdsGdsGdsAds
41
4966
|
GdsGdsGksAksAk
|
|
1213105
1917
1932
7305
7320
AksAksTksmCdsAysmCdsmCdsAdsAdsGds
21
4956
|
GdsAdsGdsGksGksAk
|
|
1213106
1918
1933
7306
7321
GksAksAksTdsCysAdsmCdsmCdsAdsAdsGds
23
4965
|
GdsAdsGksGksGk
|
|
1213107
1919
1934
7307
7322
GksGksAksAdsUysmCdsAdsmCdsmCdsAds
18
4995
|
AdsGdsGdsAksGksGk
|
|
1213108
1920
1935
7308
7323
mCksGksGksAdsAysTdsmCdsAdsmCdsmCds
28
3303
|
AdsAdsGdsGksAksGk
|
|
1213109
1921
1936
7309
7324
GksmCksGksGdsAysAdsTdsmCdsAdsmCds
37
3379
|
CdsAdsAdsGksGksAk
|
|
1213110
1922
1937
7310
7325
TksGksmCksGdsGysAdsAdsTdsmCdsAdsm
20
3455
|
CdsmCdsAdsAksGksGk
|
|
1213111
1923
1938
7311
7326
mCksTksGksmCdsGysGdsAdsAdsTdsCds
12
3531
|
AdsmCdsmCdsAksAksGk
|
|
1213112
1924
1939
7312
7327
AksmCksTksGdsCysGdsGdsAdsAdsTdsmCds
20
3607
|
AdsmCdsmCksAksAk
|
|
1213113
1925
1940
7313
7328
mCksAksmCksTdsGysmCdsGdsGdsAdsAds
29
3683
|
TdsmCdsAdsmCksmCksAk
|
|
1213114
1926
1941
7314
7329
TksmCksAksmCdsUysGdsmCdsGdsGdsAds
30
4996
|
AdsTdsmCdsAksmCksmCk
|
|
1213115
1927
1942
7315
7330
mCksTksmCksAdsCysTdsGdsmCdsGdsGds
18
3835
|
AdsAdsTdsmCksAksmCk
|
|
1213116
1928
1943
7316
7331
TksmCksTksmCdsAysmCdsTdsGdsmCdsGds
28
3911
|
GdsAdsAdsTksmCksAk
|
|
1213117
1929
1944
7317
7332
mCksTksmCksTdsCysAdsmCdsTdsGdsmCds
26
4971
|
GdsGdsAdsAksTksmCk
|
|
1213118
1930
1945
7318
7333
TksmCksTksmCdsUysmCdsAdsmCdsTdsGds
19
4997
|
mCdsGdsGdsAksAksTk
|
|
1213119
1931
1946
7319
7334
mCksTksmCksTdsCysTdsmCdsAdsmCdsTds
52
2461
|
GdsmCdsGdsGksAksAk
|
|
1213120
1932
1947
7320
7335
AksmCksTksmCdsUysmCdsTdsmCdsAdsm
10
4998
|
CdsTdsGdsmCdsGksGksAk
|
|
1213121
1933
1948
7321
7336
mCksAksmCksTdsCysTdsmCdsTdsmCdsAds
34
4957
|
mCdsTdsGdsmCksGksGk
|
|
1213122
1934
1949
7322
7337
mCksmCksAksmCdsUysmCdsTdsmCdsTdsm
19
4999
|
CdsAdsmCdsTdsGksmCksGk
|
|
1213123
1935
1950
7323
7338
GksmCksmCksAdsCysTdsmCdsTdsmCdsTds
33
4967
|
mCdsAdsmCdsAksGksmCk
|
|
1213124
1936
1951
7324
7339
AksGksmCksmCdsAysmCdsTdsmCdsTdsm
14
4961
|
CdsTdsmCdsAdsmCksTksGk
|
|
1213125
1937
1952
7325
7340
mCksAksGksmCdsCysAdsmCdsTdsmCdsTds
42
4972
|
mCdsTdsmCdsAksmCksTk
|
|
1213126
2000
2015
7388
7403
mCksGksmCksmCdsAysTdsmCdsmCdsTds
53
3228
|
GdsGdsmCdsGdsmCksGksGk
|
|
1213127
2001
2016
7389
7404
GksmCksGksmCdsCysAdsTdsmCdsmCdsTds
34
4964
|
GdsGdsmCdsGksmCksGk
|
|
1213128
2002
2017
7390
7405
TksGksmCksGdsCysmCdsAdsTdsmCdsmCds
61
3305
|
TdsGdsGdsmCksGksmCk
|
|
1213129
2003
2018
7391
7406
mCksTksGksmCdsGysmCdsmCdsAdsTdsm
114
3381
|
CdsmCdsTdsGdsGksmCksGk
|
|
1213130
2004
2019
7392
7407
mCksmCksTksGdsCysGdsmCdsmCdsAdsTds
24
3457
|
mCdsmCdsTdsGksGksmCk
|
|
1213131
2005
2020
7393
7408
TksmCksmCksTdsGysmCdsGdsmCdsmCds
35
3533
|
AdsTdsmCdsmCdsTksGksGk
|
|
1213132
2006
2021
7394
7409
TksTksmCksmCdsCysGdsmCdsGdsmCdsm
29
5000
|
CdsAdsTdsmCdsmCksTksGk
|
|
1213133
2007
2022
7395
7410
GksTksTksmCdsCysTdsGdsmCdsGdsmCdsm
33
3685
|
CdsAdsTdsmCksmCksTk
|
|
1213134
2008
2023
7396
7411
AksGksTksTdsCysmCdsTdsGdsmCdsGdsm
33
3761
|
CdsmCdsAdsTksmCksmCk
|
|
1213135
2009
2024
7397
7412
GksAkGGksTdsUysmCdsmCdsTdsGdsmCds
9
5001
|
GdsmCdsmCdsAksTksmCk
|
|
1213136
2010
2025
7398
7413
TksGksAksGdsUysTdsmCdsmCdsTdsGdsm
20
5002
|
CdsGdsmCdsmCksAksTk
|
|
1213137
2011
2026
7399
7414
TksTksGksAdsGysTdsTdsmCdsmCdsTdsGds
17
2463
|
mCdsGdsmCksmCksAk
|
|
1213138
2012
2027
7400
7415
AksTksTksGdsAysGdsTdsTdsmCdsmCdsTds
17
2540
|
GdsmCdsGksmCksmCk
|
|
1213139
2013
2028
7401
7416
TksAksTksTdsGysAdsGdsTdsTdsmCdsmCds
26
2617
|
TdsGdsmCksCksCk
|
|
1213140
2014
2029
7402
7417
TksTksAksTdsUysGdsAdsGdsTdsTdsmCds
25
5003
|
CdsTdsGksmCksGk
|
|
1213141
2016
2031
7404
7419
mCksTksTksTdsAysTdsTdsGdsAdsGdsTds
22
2771
|
TdsmCdsmCksTksGk
|
|
1213142
2017
2032
7405
7420
AksmCksTksTdsUysAdsTdsTdsGdsAdsGds
30
5004
|
TdsTdsmCksmCksTk
|
|
1213143
2018
2033
7406
7421
mCksAksmCksTdsUysTdsAdsTdsTdsGdsAds
25
5005
|
GdsTdsTksmCksmCk
|
|
1213144
2019
2034
7407
7422
GksmCksAksmCdsUysTdsTdsAdsTdsTdsGds
10
5006
|
AdsGdsTksTksmCk
|
|
1213145
2020
2035
7408
7423
AksGksmCksAdsCysTdsTdsTdsAdsTdsTds
20
3076
|
GdsAdsGksTksTk
|
|
1213146
2021
2036
7409
7424
AksAksGksmCdsAysmCdsTdsTdsTdsAdsTds
11
3147
|
TdsGdsAksGksTk
|
|
1213147
2022
2037
7410
7425
AksAksAksGdsCysAdsmCdsTdsTdsTdsAds
15
3229
|
TdsTdsGksAksGk
|
|
1213148
2023
2038
7411
7426
mCksAksAksAdsGysmCdsAdsmCdsTdsTds
34
3306
|
TdsAdsTdsTksGksAk
|
|
1213149
2024
2039
7412
7427
TksmCksAksAdsAysGdsmCdsAdsmCdsTds
35
3382
|
TdsTdsAdsTksTksGk
|
|
1213150
2031
2046
7419
7434
AksGksmCksAdsUysTdsTdsTdsmCdsAdsAds
41
5007
|
AdsGdsmCksAksmCk
|
|
1213151
2033
2048
7421
7436
TksmCksAksGdsCysAdsTdsTdsTdsTdsmCds
23
3534
|
AdsAdsAksGksmCk
|
|
TABLE 88
|
|
Reduction of FXII RNA (Huh7, electroporation, 2000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
|
Compound
Start
Stop
Start
Stop
FXII
SEQ ID
|
Number
Site
Site
Site
Site
Chemistry notation (5′ to 3′)
(%UTC)
NO
|
|
1128769
153
168
N/A
N/A
AksGksAksAdsmCdsGdsAdsCdsTdsGds
41
3723
|
TdsGdsTdsGksmCksTk
|
|
1128802
218
233
3569
3584
TksAksmCksAdsTdsTdsTdsGdsTdsGdsGds
N/A
2426
|
TdsAdsmCksAksGk
|
|
1128870
470
485
4415
4430
TksGksTksGdsGdsAdsAdsAdsAdsAdsmCds
31
3728
|
mCdsGdsGksAksGk
|
|
1129491
1927
1942
7315
7330
mCksTksmCksAdsmCdsTdsGdsmCdsGdsGds
16
3835
|
AdsAdsTdsmCksAksmCk
|
|
1129510
1984
1999
7372
7387
AksGksmCksTdsGdsGdsmCdsmCdsGdsmCds
29
3760
|
AdsmCdsTdsGksGksGk
|
|
1129530
2008
2023
7396
7411
AksGksTksTdsmCdsmCdsTdsGdsmCdsGdsm
80
3761
|
CdsmCdsAdsTksmCksmCk
|
|
1129531
2009
2024
7397
7412
GksAksGksTdsTdsmCdsmCdsTdsGdsmCds
29
3837
|
GdsmCdsmCdsAksTksmCk
|
|
1129532
2010
2025
7398
7413
TksGksAksGdsTdsTdsmCdsmCdsTdsGdsm
19
3913
|
CdsGdsmCdsmCksAksTk
|
|
1129540
2019
2034
7407
7422
GksmCksAksmCdsTdsTdsTdsAdsTdsTdsGds
48
3000
|
AdsGdsTksTksmCk
|
|
1129683
N/A
N/A
1003
1018
TksmCksAksAdsmCdsmCdsTdsGdsGdsTds
64
3236
|
TdsTdsmCdsAksmCksAk
|
|
1129703
N/A
N/A
1139
1154
AksTksGksGadsmCdsAdsGdsGdsTdsAdsm
32
3237
|
CdsmCdsmCdsTksTksCk
|
|
1129715
N/A
N/A
1559
1574
CksAksGksGdsGdsTdsAdsAdsGdsGdsTds
52
2626
|
mCdsmCdsAksTksCk
|
|
1129783
N/A
N/A
2494
2509
GksTksTksAdsGdsAdsmCdsAdsmCdsTdsTds
104
3241
|
GdsAdsTksTksmCk
|
|
1129796
N/A
N/A
2628
2643
TksmCksTksAdsmCdsTdsTdsAdsGdsTdsGds
57
2707
|
mCdsAdsAksmCksGk
|
|
1129877
N/A
N/A
4035
4050
GksmCksTksAdsCdsTdsmCdsmCdsAdsAds
66
2788
|
CdsTdsTdsTksmCksmCk
|
|
1129916
N/A
N/A
4363
4378
GksGksGksAdsTdsAdsGdsTdsGdsGdsTdsm
103
2713
|
CdsTdsmCksAksGk
|
|
1129938
N/A
N/A
4635
4650
AksmCksGksmCdsAdsGdsTdsGdsAdsGdsm
52
2868
|
CdsmCdsAdsmCksmCksmCk
|
|
1129943
N/A
N/A
4808
4823
GksGksmCksAdsAdsGdsmCdsmCdsmCdsGds
63
3249
|
TdsmCdsmCdsmCksAksmCk
|
|
1129948
N/A
N/A
4840
4855
AksTksmCksTdsGdsmCdsTdsTdsTdsmCdsm
108
3630
|
CdsGdsmCdsAksmCksTk
|
|
1129949
N/A
N/A
4844
4859
TksmCksTksmCdsAdsTdsmCdsTdsGdsmCds
81
3706
|
TdsTdsTdsmCksmCksGk
|
|
1213152
141
156
508
523
TksGksmCksTdsmCdsUysTdsmCdsAdsGdsm
72
5008
|
CdsTdsTdsTksGksTk
|
|
1213153
142
157
509
524
GksTksGksmCdsTdsCysTdsTdsmCdsAdsGds
29
4958
|
CdsTdsTksTksGk
|
|
1213154
143
158
510
525
TksGksTksGdsmCdsUysmCdsTdsTdsmCdsAds
75
5009
|
GdsmCdsTksTksTk
|
|
1213155
144
159
511
526
GksTksGksTdsGdsCysTdsmCdsTdsTdsmCds
70
4951
|
AdsGdsmCksTksTk
|
|
1213156
145
160
512
527
TksGksTksGdsTdsGysmCdsTdsmCdsTdsTds
35
3112
|
mCdsAdsGksmCksTk
|
|
1213157
146
161
513
528
mCksTksGksTdsGdsUysGdsmCdsTdsmCds
60
5010
|
TdsTdsmCdsAksGksmCk
|
|
1213158
147
162
514
529
AksmCksTksGdsTdsGysTdsGdsmCdsTdsm
56
3267
|
CdsTdsTdsmCksAksGk
|
|
1213160
149
164
516
531
mCksGksAksmCdsTdsGysTdsGdsTdsGdsm
61
3419
|
CdsTdsmCdsTksTksmCk
|
|
1213161
150
165
N/A
N/A
AksmCksGksAdsmCdsUysGdsTdsGdsTds
36
5011
|
GdsmCdsTdsmCksTksTk
|
|
1213162
151
166
N/A
N/A
AksAksmCksGdsAdsCysTdsGdsTdsGdsTds
97
3571
|
GdsmCdsTksmCksTk
|
|
1213163
152
167
N/A
N/A
GksAksAksmCdsGdsAysmCdsTdsGdsTdsGds
50
3647
|
TdsGdsmCksTksmCk
|
|
1213164
153
168
N/A
N/A
AksGksAksAdsmCdsGysAdsmCdsTdsGdsTds
99
3723
|
GdsTdsGksmCksTk
|
|
1213165
154
169
N/A
N/A
GksAksGksAdsAdsCysGdsAdsmCdsTdsGds
132
3799
|
TdsGdsTksGksmCk
|
|
1213166
155
170
N/A
N/A
TksGksAksGdsAdsAysmCdsGdsAdsmCdsTds
70
3875
|
GdsTdsGksTksGk
|
|
1213167
156
171
N/A
N/A
GksTksGksAdsGdsAysAdsmCdsGdsAdsmCds
45
2424
|
TdsGdsTksGksTk
|
|
1213168
157
172
N/A
N/A
AksGksTksGdsAdsGysAdsAdsmCdsGdsAds
112
2502
|
mCdsTdsGksTksGk
|
|
1213169
158
173
N/A
N/A
mCksAksGksTdsGdsAysGdsAdsAdsmCdsGds
73
2579
|
AdsmCdsTksGksTk
|
|
1213170
208
223
3559
3574
GksTksAksmCdsAdsGysmCdsTdsGdsmCdsm
47
4968
|
CdsGdsGdsTksGksGk
|
|
1213171
209
224
3560
3575
GksGksTksAdsmCdsAysGdsmCdsTdsGdsm
22
4952
|
CdsmCdsGdsGksTksGk
|
|
1213173
211
226
3562
3577
GksTksGksGdsTdsAysmCdsAdsGdsmCdsTds
112
4973
|
GdsmCdsmCksGksGk
|
|
1213174
212
227
3563
3578
TksGksTksGdsGdsUysAdsmCdsAdsGdsmCds
72
5012
|
TdsGdsmCksmCksGk
|
|
1213175
213
228
3564
3579
TksTksGksTdsGdsGysTdsAdsmCdsAdsGds
46
2811
|
mCdsTdsGksmCksmCk
|
|
1213176
214
229
3565
3580
TksTksTksGdsTdsGysGdsTdsAdsmCdsAds
62
2887
|
GdsmCdsTksGksmCk
|
|
1213177
215
230
3566
3581
AksTksTksTdsGdsUysGdsGdsTdsAdsmCds
45
5013
|
AdsGdsmCksTksGk
|
|
1213178
216
231
3567
3582
mCksAksTksTdsTdsGysTdsGdsGdsTdsAds
33
3039
|
mCdsAdsGksmCksTk
|
|
1213179
217
232
3568
3583
AksmCksAksTdsTdsUysGdsTdsGdsGdsTds
49
5014
|
AdsmCdsAkGksmCk
|
|
1213180
218
233
3569
3584
TksAksmCksAdsTdsUysTdsGdsTdsGdsGds
26
5015
|
TdsAdsmCksAksGk
|
|
1213181
219
234
3570
3585
GksTksAksmCdsAdsUysTdsTdsGdsTdsGds
50
5016
|
GdsTdsAksmCksAk
|
|
1213182
220
235
3571
3586
GksGksTksAdsmCdsAysTdsTdsTdsGdsTds
46
3269
|
GdsGdsTksAksmCk
|
|
1213183
221
236
3572
3587
GksGksGksTdsAdsCysAdsTdsTdsTdsGds
117
3345
|
TdsGdsGksTksAk
|
|
1213184
222
237
3573
3588
TksGksGksGdsTdsAysmCdsAdsTdsTdsTds
N/A
3421
|
GdsTdsGksGksTk
|
|
1213185
223
238
3574
3589
GksTksGksGdsGdsUysAdsmCdsAdsTdsTds
63
5017
|
TdsGdsTksGksGk
|
|
1213186
1776
1791
7164
7179
TksGksmCksAdsGdsGysGdsTdsGdsAdsGds
149
3299
|
mCdsmCdsGksGksmCk
|
|
1213187
1777
1792
7165
7180
TksTksGksmCdsAdsGysGdsGdsTdsGdsAds
98
3375
|
GdsmCdsmCksGksGk
|
|
1213188
1778
1793
7166
7181
mCksTksTksGdsmCdsAysGdsGdsGdsTdsGds
71
4962
|
AdsGdsmCksmCksGk
|
|
1213189
1779
1794
7167
7182
mCksmCksTksTdsGdsCysAdsGdsGdsGdsTds
40
4974
|
GdsAdsGksmCksmCk
|
|
1213190
1780
1795
7168
7183
GksmCksmCksTdsTdsGysmCdsAdsGdsGds
53
3451
|
GdsTdsGdsAksGksmCk
|
|
1213192
1782
1797
7170
7185
AksTksGksmCdsmCdsUysTdsGdsmCdsAds
107
5018
|
GdsGdsGdsTksGksAk
|
|
1213193
1783
1798
7171
7186
GksAksTksGdsmCdsCysTdsTdsGdsmCdsAds
54
4959
|
GdsGdsGksTksGk
|
|
1213194
1784
1799
7172
7187
TksGksAksTdsGdsCysmCdsTdsTdsGdsmCds
28
4969
|
AdsGdsGksGksTk
|
|
1213195
1785
1800
7173
7188
AksTksGksAdsTdsGysmCdsmCdsTdsTdsGds
53
3603
|
mCdsAdsGksGksGk
|
|
1213196
1786
1801
7174
7189
GksAksTksGdsAdsUysGdsmCdsmCdsTdsTds
73
5019
|
GdsmCdsAksGksGk
|
|
1213197
1787
1802
7175
7190
TksGksAksTdsGdsAysTdsGdsmCdsmCdsTds
43
4975
|
TdsGdsmCksAksGk
|
|
1213198
1788
1803
7176
7191
mCksTksGksAdsTdsGysAdsTdsGdsmCdsm
65
3679
|
CdsTdsTdsGksmCksAk
|
|
1213199
1789
1804
7177
7192
GksmCksTksGdsAdsUysGdsAdsTdsGdsm
80
5020
|
CdsmCdsTdsTksGksmCk
|
|
1213200
1790
1805
7178
7193
AksGksmCksTdsGdsAysTdsGdsAdsTdsGds
49
3831
|
mCdsmCdsTksTksGk
|
|
1213201
1791
1806
7179
7194
mCksAksGksmCdsTdsGysAdsTdsGdsAdsTds
83
4960
|
GdsmCdsmCksTksTk
|
|
1213202
1792
1807
7180
7195
mCksmCksAksGdsmCdsUysGdsAdsTdsGds
44
5021
|
AdsTdsGdsmCksmCksTk
|
|
1213203
1793
1808
7181
7196
mCksmCksmCksAdsGdsCysTdsGdsAdsTds
42
4953
|
GdsAdsTdsGksmCksmCk
|
|
1213204
1892
1907
7280
7295
mCksmCksmCksTdsGdsAysGdsmCdsAdsAds
53
3530
|
TdsmCdsAdsGksGksAk
|
|
1213205
1893
1908
7281
7296
TksmCksmCksmCdsTdsGysAdsGdsmCdsAds
104
3606
|
AdsTdsmCdsAksGksGk
|
|
1213206
1894
1909
7282
7297
GksTksmCksmCdsmCdsUysGdsAdsGdsmCds
45
5022
|
AdsAdsTdsmCksAksGk
|
|
1213207
1895
1910
7283
7298
AksGksTksmCdsmCdsCysTdsGdsAdsGdsm
36
3758
|
CdsAdsAdsTksCksAk
|
|
1213208
1896
1911
7284
7299
GksAkGksTdsmCdsCysmCdsTdsGdsAds
62
3834
|
GdsmCdsAdsAksTksmCk
|
|
1213209
1897
1912
7285
7300
TksGksAksGdsTdsCysmCdsmCdsTdsGds
46
3910
|
AdsGdsmCdsAksAksTk
|
|
TABLE 89
|
|
Reduction of FXII RNA (Huh7, electroporation, 2000 nM modified oligonucleotide)
|
SEQ
SEQ
SEQ
SEQ
|
ID
ID
ID
ID
|
NO:
NO:
NO:
NO:
|
1
1
2
2
|
Compound
Start
Stop
Start
Stop
Sequence and Chemistry notation
FXII
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
(%UTC)
NO
|
|
1128769
153
168
N/A
N/A
AksGksAksAdsmCdsGdsAdsmCdsTdsGdsTds
84
3723
|
GdsTdsGksmCksTk
|
|
1128796
212
227
3563
3578
TksGksTksGdsGdsTdsAdsmCdsAdsGdsmCds
103
2734
|
TdsGdsmCksmCksGk
|
|
1128802
218
233
3569
3584
TksAksmCksAdsTdsTdsTdsGdsTdsGdsGds
97
2426
|
TdsAdsmCksAksGk
|
|
1128870
470
485
4415
4430
TksGksTksGdsGdsAdsAdsAdsAdsAdsCds
60
3728
|
mCdsGdsGksAksGk
|
|
1129221
1357
1372
6017
6032
mCksGksTksmCdsTdsGdsGdsmCdsAdsmCds
19
3060
|
GdsGdsmCdsTksmCksAk
|
|
1129407
1781
1796
7169
7184
TksGksmCksmCdsTdsTdsGdsmCdsAdsGds
12
3527
|
GdsGdsTdsGksAksGk
|
|
1129472
1897
1912
7285
7300
TksGksAksGdsTdsmCdsmCdsmCdsTdsGds
18
3910
|
AdsGdsmCdsAksAksTk
|
|
1129488
1924
1939
7312
7327
AksmCksTksGdsmCdsGdsGdsAdsAdsTdsm
5
3607
|
CdsAdsmCdsmCksAksAk
|
|
1129491
1927
1942
7315
7330
mCksTksmCksAdsCdsTdsGdsmCdsGdsGds
15
3835
|
AdsAdsTdsmCksAksmCk
|
|
1129531
2009
2024
7397
7412
GksAksGksTdsTdsmCdsmCdsTdsGdsmCds
12
3837
|
GdsmCdsmCdsAksTksmCk
|
|
1129532
2010
2025
7398
7413
TksGksAksGdsTdsTdsmCdsmCdsTdsGdsm
11
3913
|
CdsGdsmCdsmCksAksTk
|
|
1129533
2011
2026
7399
7414
TksTksGksAdsGdsTdsTdsmCdsmCdsTdsGds
8
2463
|
mCdsGdsmCksmCksAk
|
|
1129540
2019
2034
7407
7422
GksmCksAksmCdsTdsTdsTdsAdsTdsTdsGds
4
3000
|
AdsGdsTksTksmCk
|
|
1129542
2021
2036
7409
7424
AksAksGksmCdsAdsmCdsTdsTdsTdsAdsTds
17
3147
|
TdsGdsAksGksTk
|
|
1129565
N/A
N/A
144
159
AksTksAksGdsmCdsGdsAdsmCdsmCdsmCds
82
3383
|
mCdsmCdsmCdsAksGksAk
|
|
1129744
N/A
N/A
2082
2097
TksGksGksAdsmCdsmCdsTdsTdsTdsAdsAds
75
3316
|
GdsAdsTksGksmCk
|
|
1129864
N/A
N/A
3897
3912
TksGksGksTdsGdsAdsTdsAdsmCdsmCdsAds
74
3322
|
GdsGdsAksGksAk
|
|
1129946
N/A
N/A
4812
4827
TksmCksmCksTdsGdsGdsmCdsAdsAdsGdsm
119
3478
|
CdsmCdsmCdsGksTksmCk
|
|
1129948
N/A
N/A
4840
4855
AksTksmksTdsGdsmCdsTdsTdsTdsmCdsm
77
3630
|
CdsGdsmCdsAksmCksTk
|
|
1129949
N/A
N/A
4844
4859
TksmCksTksmCdsAdsTdsmCdsTdsGdsmCds
84
3706
|
TdsTdsTdsmCksmCksGk
|
|
1213210
1898
1913
7286
7301
AksTksGksAdsGdsUysCdsmCdsmCdsTds
40
5023
|
GdsAdsGdsmCksAksAk
|
|
1213211
1899
1914
7287
7302
GksAksTksGdsAdsGysTdsmCdsmCdsmCds
27
2537
|
TdsGdsAdsGksmCksAk
|
|
1213212
1900
1915
7288
7303
AksGksAksTdsGdsAysGdsTdsmCdsmCdsm
24
2614
|
CdsTdsGdsAksGksmCk
|
|
1213213
1901
1916
7289
7304
AksAksGksAdsTdsGysAdsGdsTdsmCdsm
19
2691
|
CdsmCdsTdsGksAksGk
|
|
1213214
1902
1917
7290
7305
AksAksAksGdsAdsUysGdsAdsGdsTdsmCds
31
5024
|
mCdsmCdsTksGksAk
|
|
1213215
1903
1918
7291
7306
GksAksAksAdsGdsAysTdsGdsAdsGdsTdsm
23
2845
|
CdsmCdsmCksTksGk
|
|
1213216
1904
1919
7292
7307
GksGksAksAdsAdsGysAdsTdsGdsAdsGds
39
2921
|
TdsmCdsmCksmCksTk
|
|
1213217
1905
1920
7293
7308
GksGksGksAdsAdsAysGdsAdsTdsGdsAds
55
2997
|
GdsTdsmCksmCksmCk
|
|
1213218
1906
1921
7294
7309
AksGksGksGdsAdsAysAdsGdsAdsTdsGds
18
3073
|
AdsGdsTksmCksmCk
|
|
1213219
1914
1929
7302
7317
mCksAksmCksmCdsAdsAysGdsGdsAdsGds
31
4954
|
GdsGdsAdsAksAksGk
|
|
1213220
1915
1930
7303
7318
TksmCksAksmCdsmCdsAysAdsGdsGdsAds
9
4948
|
GdsGdsGdsAksAksAk
|
|
1213221
1916
1931
7304
7319
AksTksmCksAdsmCdsCysAdsAdsGdsGdsAds
31
4966
|
GdsGdsGksAksAk
|
|
1213222
1917
1932
7305
7320
AksAksTksmCdsAdsCysmCdsAdsAdsGdsGds
44
4956
|
AdsGdsGksGksAk
|
|
1213223
1918
1933
7306
7321
GksAksAksTdsmCdsAysmCadsmCdsAdsAds
28
4965
|
GdsGdsAdsGksGksGk
|
|
1213224
1919
1934
7307
7322
GksGksAksAdsTdsCysAdsmCdsmCdsAdsAds
21
3226
|
GdsGdsAksGksGk
|
|
1213225
1920
1935
7308
7323
mCksGksGksAdsAdsUysmCdsAdsmCdsmCds
33
5025
|
AdsAdsGdsGksAksGk
|
|
1213226
1921
1936
7309
7324
GksmCksGksGdsAdsAysTdsmCdsAdsmCds
21
3379
|
mCdsAdsAdsGksGksAk
|
|
1213227
1922
1937
7310
7325
TksGksmCksGdsGdsAysAdsTdsmCdsAdsm
8
3455
|
CdsmCdsAdsAksGksGk
|
|
1213228
1923
1938
7311
7326
mCksTksGksmCdsGdsGysAdsAdsTdsmCds
17
3531
|
AdsmCdsmCdsAksAksGk
|
|
1213229
1924
1939
7312
7327
AksmCksTksGdsmCdsGysGdsAdsAdsTdsm
12
3607
|
CdsAdsmCdsmCksAksAk
|
|
1213230
1925
1940
7313
7328
mCksAksmCksTdsGdsCysGdsGdsAdsAdsTds
22
3683
|
mCdsAdsmCksmCksAk
|
|
1213231
1926
1941
7314
7329
TksmCksAksmCdsTdsGysmCdsGdsGdsAds
23
3759
|
AdsTdsmCdsAksmCksmCk
|
|
1213232
1927
1942
7315
7330
mCksTksmCksAdsmCdsUysGdsmCdsGdsGds
11
5026
|
AdsAdsTdsmCksAksmCk
|
|
1213233
1928
1943
7316
7331
TksmCksTksmCdsAdsCysTdsGdsmCdsGds
16
3911
|
GdsAdsAdsTksmCksAk
|
|
1213234
1929
1944
7317
7332
mCksTksmCksTdsmCdsAysmCdsTdsGdsmCds
11
4971
|
GdsGdsAdsAksTksmCk
|
|
1213235
1930
1945
7318
7333
TksmCksTksmCdsTdsCysAdsmCdsTdsGdsm
11
4955
|
CdsGdsGdsAksAksTk
|
|
1213236
1931
1946
7319
7334
mCksTksmCksTdsmCdsUysmCdsAdsmCdsTds
34
5027
|
GdsmCdsGdsGksAksAk
|
|
1213237
1932
1947
7320
7335
AksmCksTksmCdsTdsCysTdsmCdsAdsmCds
20
4976
|
TdsGdsmCdsGksGksAk
|
|
1213238
1933
1948
7321
7336
mCksAksmCksTdsmCdsUysmCdsTdsmCds
26
5028
|
AdsmCdsTdsGdsmCksGksGk
|
|
1213239
1934
1949
7322
7337
mCksmCksAksmCdsTdsCysTdsmCdsTdsmCds
18
4949
|
AdsmCdsTdsGksmCksGk
|
|
1213240
1935
1950
7323
7338
GksmCksmCksAdsmCdsUysmCdsTdsmCds
32
5029
|
TdsmCdsAdsmCdsTksGksmCk
|
|
1213241
1936
1951
7324
7339
AksGksmCksmCdsAdsCysTdsmCdsTdsmCds
28
4961
|
TdsmCdsAdsmCksTksGk
|
|
1213242
1937
1952
7325
7340
mCksAksGksmCdsmCdsAysmCdsTdsmCds
55
4972
|
TdsmCdsTdsmCdsAksmCksTk
|
|
1213243
2000
2015
7388
7403
mCksGksmCksmCdsAdsUysmCdsmCdsTds
92
5030
|
GdsGdsmCdsGdsmCksGksGk
|
|
1213244
2001
2016
7389
7404
GksmCksGksmCdsmCdsAysTdsmCdsmCds
92
4964
|
TdsGdsGdsmCdsGksmCksGk
|
|
1213245
2002
2017
7390
7405
TksGksmCksGdsmCdsCysAdsTdsmCdsmCds
71
3305
|
TdsGdsGdsmCksGksmCk
|
|
1213246
2003
2018
7391
7406
mCksTksGksmCdsGdsCysmCdsAdsTdsmCds
54
3381
|
mCdsTdsGdsGksmCksGk
|
|
1213247
2004
2019
7392
7407
mCksmCksTksGdsmCdsGysmCdsmCdsAds
54
3457
|
TdsmCdsmCdsTdsGksGksmCk
|
|
1213248
2005
2020
7393
7408
TksmCksmCksTdsGdsCysGdsmCdsmCdsAds
21
3533
|
TdsmCdsmCdsTksGksGk
|
|
1213249
2006
2021
7394
7409
TksTksmCksmCdsTdsGysmCdsGdsmCdsmCds
64
3609
|
AdsTdsmCdsmCksTksGk
|
|
1213250
2007
2022
7395
7410
GksTksTksmCdsmCdsUysGdsmCdsGdsmCds
30
5031
|
mCdsAdsTdsmCksmCksTk
|
|
1213251
2008
2023
7396
7411
AksGksTksTdsmCdsCysTdsGdsmCdsGdsm
21
3761
|
CdsmCdsAdsTksmCksmCk
|
|
1213252
2009
2024
7397
7412
GksAksGksTdsTdsCysmCdsTdsGdsmCdsGds
24
3837
|
mCdsmCdsAksTksmCk
|
|
1213253
2010
2025
7398
7413
TksGksAksGdsTdsUysmCdsmCdsTdsGdsm
17
5032
|
CdsGdsmCdsmCksAksTk
|
|
1213254
2011
2026
7399
7414
TksTksGksAdsGdsUysTdsmCdsmCdsTdsGds
30
5033
|
mCdsGdsmCksmCksAk
|
|
1213255
2012
2027
7400
7415
AksTksTksGdsAdsGysTdsTdsmCdsmCdsTds
20
2540
|
GdsmCdsGksmCksmCk
|
|
1213256
2013
2028
7401
7416
TksAksTksTdsGdsAysGdsTdsTdsmCdsmCds
26
2617
|
TdsGdsmCksGksmCk
|
|
1213257
2014
2029
7402
7417
TksTksAksTdsTdsGysAdsGdsTdsTdsmCdsm
21
2694
|
CdsTdsGksmCksGk
|
|
1213258
2016
2031
7404
7419
mCksTksTksTdsAdsUysTdsGdsAdsGdsTds
19
5034
|
TdsmCdsmCksTksGk
|
|
1213259
2017
2032
7405
7420
AksmCksTksTdsTdsAysTdsTdsGdsAdsGds
2
2848
|
TdsTdsmCksmCksTk
|
|
1213260
2018
2033
7406
7421
mCksAksmCksTdsTdsUysAdsTdsTdsGdsAds
2
5035
|
GdsTdsTksmCksmCk
|
|
1213261
2019
2034
7407
7422
GksmCksAksmCdsTdsUysTdsAdsTdsTdsGds
12
5036
|
AdsGdsTksTksmCk
|
|
1213262
2020
2035
7408
7423
AksGksmCksAdsmCdsUysTdsTdsAdsTdsTds
4
5037
|
GdsAdsGksTksTk
|
|
1213263
2021
2036
7409
7424
AksAksGksmCdsAdsCysTdsTdsTdsAdsTds
19
3147
|
TdsGdsAksGksTk
|
|
1213264
2022
2037
7410
7425
AksAksAksGdsmCdsAysmCdsTdsTdsTdsAds
11
3229
|
TdsTdsGksAksGk
|
|
1213265
2023
2038
7411
7426
mCksAksAksAdsGdsCysAdsmCdsTdsTdsTds
19
3306
|
AdsTdsTksGksAk
|
|
1213266
2024
2039
7412
7427
TksmCksAksAdsAdsGysmCdsAdsmCdsTds
54
3382
|
TdsTdsAdsTksTksGk
|
|
1213267
2031
2046
7419
7434
AksGksmCksAdsTdsUysTdsTdsmCdsAdsAds
23
5038
|
AdsGdsmCksAksmCk
|
|
1213268
2033
2048
7421
7436
TksmCksAksGdsmCdsAysTdsTdsTdsTdsm
20
3534
|
CdsAdsAdsAksGksCk
|
|
Example 5: Dose-Dependent Reduction of Human FXII RNA by Modified Oligonucleotides In Vitro
Certain modified oligonucleotides described in the studies above exhibiting significant in vitro inhibition of FXII RNA were selected and tested at various doses in HepG2 cells.
The modified oligonucleotides were tested in a series of experiments using the same culture conditions. The results for each experiment are presented in separate tables shown below. Cultured HepG2 cells at a density of 10,000 cells per well were transfected using lipofectin with modified oligonucleotides diluted to different concentrations as specified in the tables below. After a treatment period of approximately 24 hours, FXII RNA levels were measured as previously described using the Human FXII primer-probe set RTS2976_MGB (forward sequence CCGGGAGCACACCGTTT, designated herein as SEQ ID NO: 11; reverse sequence GGAATCACCAAGGAGGGAAAG, designated herein as SEQ ID NO: 12; probe sequence CTGATTGCTCAGGGACT, designated herein as SEQ ID NO: 13). FXII RNA levels were normalized to total RNA, as measured by RIBOGREEN®. Reduction of FXII RNA is presented in the tables below as percent FXII RNA relative to the amount in untreated control cells (% UTC).
The half maximal inhibitory concentration (IC50) of each modified oligonucleotide was calculated using a linear regression on a log/linear plot of the data in excel and is also presented in the tables below.
TABLE 90
|
|
Dose-dependent reduction of human FXII RNA by
|
modified oligonucleotides
|
Compound
FXII (% UTC)
150
IC50
|
Number
5 nM
9 nM
19 nM
38 nM
75 nM
nM
(μM)
|
|
413627
74
57
45
30
22
18
0.02
|
413631
77
66
46
36
27
11
0.02
|
413632
80
64
53
42
27
21
0.02
|
413636
87
71
58
44
28
21
0.03
|
413637
85
66
56
36
24
15
0.02
|
413642
72
57
42
32
17
11
0.01
|
413643
75
50
37
25
18
13
0.01
|
413644
81
77
54
39
29
18
0.03
|
413645
80
69
51
35
28
21
0.02
|
413652
77
62
46
32
21
12
0.02
|
|
Example 6: Dose-Dependent Reduction of Human FXII RNA by Modified Oligonucleotides In Vitro
Certain modified oligonucleotides described in the studies above exhibiting significant in vitro inhibition of FXII RNA were selected and tested at various doses in Huh7 cells.
The modified oligonucleotides were tested in a series of experiments using the same culture conditions. The results for each experiment are presented in separate tables shown below. Cultured Huh7 cells at a density of 5,000 cells per well were transfected using oligofectamine with modified oligonucleotides diluted to different concentrations as specified in the tables below. After a treatment period of approximately 24 hours, FXII RNA levels were measured as previously described using the Human FXII primer-probe set RTS2992. FXII RNA levels were normalized to total RNA, as measured by RIBOGREEN®. Reduction of FXII RNA is presented in the tables below as percent FXII RNA relative to the amount in untreated control cells (% UTC).
The half maximal inhibitory concentration (IC50) of each modified oligonucleotide was calculated using a linear regression on a log/linear plot of the data in excel and is also presented in the tables below. The symbol “†” indicates that the modified oligonucleotide is complementary to the target transcript within the amplicon region of the primer probe set. In such instances, additional assays using alternative primer probes must be performed to accurately assess the potency and efficacy of such modified oligonucleotides.
TABLE 91
|
|
Dose-dependent reduction of human FXII RNA
|
by modified oligonucleotides
|
FXII (% UTC)
|
Compound
14
28
55
110
220
IC50
|
Number
nM
nM
nM
nM
nM
μM
|
|
462145
87
68
48
37
20
0.06
|
462147
80
66
43
34
28
0.056
|
462176
95
79
67
56
31
0.113
|
462183
76
64
52
39
24
0.058
|
462189
56
60
37
25
22
0.028
|
462190
72
67
52
34
26
0.056
|
462192
53
44
32
15
22
0.015
|
462193
86
75
48
43
27
0.072
|
462195
50
42
35
19
23
0.013
|
462196
53
31
36
17
19
0.012
|
462197
46
40
31
17
15
<14 nM
|
462198
42
34
32
27
19
<14 nM
|
|
TABLE 92
|
|
Dose-dependent reduction of human FXII RNA
|
by modified oligonucleotides
|
FXII (% UTC)
|
Compound
14
28
55
110
220
IC50
|
Number
nM
nM
nM
nM
nM
μM
|
|
462072
80
55
40
26
12
0.041
|
462073
81
47
30
22
23
0.035
|
462098
89
89
57
38
19
0.075
|
462107
90
88
54
58
32
0.109
|
462131
69
61
68
42
34
0.08
|
462188
95
84
52
46
41
0.106
|
462194
73
63
50
40
49
0.087
|
462199
50
46
44
28
31
0.016
|
462064†
73
62
32
16
13
N/A
|
462065†
66
48
41
13
4
N/A
|
462068†
56
52
29
15
8
N/A
|
462069†
81
57
53
21
10
N/A
|
|
Example 7: Dose-Dependent Reduction of Human FXII RNA by Modified Oligonucleotides In Vitro
Certain modified oligonucleotides described in the studies above exhibiting significant in vitro inhibition of FXII RNA were selected and tested at various doses in Huh7 cells.
The modified oligonucleotides were tested in a series of experiments using the same culture conditions. The results for each experiment are presented in separate tables shown below. Cultured Huh7 cells at a density of 20,000 cells per well were transfected using electroporation with modified oligonucleotides diluted to different concentrations as specified in the tables below. After a treatment period of approximately 24 hours, FXII RNA levels were measured as previously described using the Human FXII primer-probe set RTS40528. FXII RNA levels were normalized to total RNA, as measured by RIBOGREEN®. Reduction of FXII RNA is presented in the tables below as percent FXII RNA relative to the amount in untreated control cells (% UTC).
The half maximal inhibitory concentration (IC50) of each modified oligonucleotide was calculated using a linear regression on a log/linear plot of the data in excel and is also presented in the tables below.
TABLE 93
|
|
Dose-dependent reduction of human FXII RNA
|
by modified oligonucleotides
|
Compound
FXII (% UTC)
|
Number
47 nM
188 nM
750 nM
3000 nM
IC50 μM
|
|
1128754
94
82
66
37
1.6
|
1128802
81
54
38
16
0.3
|
1128835
92
83
57
37
1.3
|
1128873
91
75
63
42
1.8
|
1128874
93
67
41
17
0.5
|
1128875
93
76
60
36
1.3
|
1129015
93
71
44
30
0.7
|
1129094
81
66
42
24
0.5
|
1129114
92
64
50
23
0.6
|
1129294
86
55
55
30
0.6
|
1129473
84
63
37
22
0.4
|
1129475
76
50
28
15
0.2
|
1129476
70
44
40
22
0.2
|
1129493
81
48
23
11
0.2
|
1129494
87
85
45
26
0.7
|
1129533
65
54
21
21
0.2
|
1129534
77
46
36
17
0.2
|
1129535
80
64
32
25
0.4
|
1129536
85
47
30
25
0.3
|
|
TABLE 94
|
|
Dose-dependent reduction of human FXII
|
RNA by modified oligonucleotides
|
FXII (% UTC)
|
Compound
47
188
750
3000
|
Number
nM
nM
nM
nM
IC50 μM
|
|
1128795
84
63
38
28
0.5
|
1128796
66
50
21
12
0.2
|
1128797
86
71
37
24
0.5
|
1128802
86
62
37
15
0.4
|
1128836
80
67
44
30
0.6
|
1128878
85
74
50
27
0.7
|
1128958
92
94
74
42
>3.0
|
1129058
94
54
45
28
0.5
|
1129136
77
68
62
32
1.0
|
1129477
96
56
39
26
0.5
|
1129478
98
65
37
22
0.5
|
1129496
88
76
70
45
2.8
|
1129497
100
75
43
17
0.6
|
1129498
80
69
43
32
0.6
|
1129499
77
65
31
26
0.4
|
1129516
82
43
38
32
0.3
|
1129537
69
57
21
16
0.2
|
1129538
55
38
18
11
0.1
|
1129539
45
22
14
7
<0.04
|
|
TABLE 95
|
|
Dose-dependent reduction of human FXII RNA
|
by modified oligonucleotides
|
FXII (% UTC)
|
Compound
47
188
750
3000
IC50
|
Number
nM
nM
nM
nM
μM
|
|
1128738
88
72
56
32
0.9
|
1128799
92
68
58
22
0.7
|
1128802
104
73
40
23
0.6
|
1128879
81
76
33
23
0.5
|
1128880
97
72
39
25
0.6
|
1128980
112
85
62
42
1.7
|
1129000
118
97
65
38
1.7
|
1129020
106
92
52
35
1.2
|
1129039
84
63
45
24
0.5
|
1129119
108
77
65
35
1.3
|
1129139
78
58
44
19
0.4
|
1129180
97
88
40
34
0.9
|
1129420
88
69
47
33
0.8
|
1129460
86
60
39
24
0.4
|
1129479
98
58
36
17
0.4
|
1129481
79
63
42
31
0.5
|
1129500
82
64
44
18
0.4
|
1129540
48
28
19
11
<0.04
|
1129541
64
36
17
10
0.1
|
|
TABLE 96
|
|
Dose-dependent reduction of human FXII RNA by modified
|
FXII (% UTC)
|
Compound
47
188
750
3000
|
Number
nM
nM
nM
nM
IC50 μM
|
|
1128762
73
63
37
21
0.3
|
1128801
99
56
35
18
0.4
|
1128802
97
65
37
14
0.4
|
1128821
78
55
57
21
0.5
|
1128841
106
91
73
36
1.9
|
1128881
80
66
42
27
0.5
|
1129022
122
100
80
46
2.9
|
1129043
112
98
57
31
1.2
|
1129142
98
79
48
20
0.7
|
1129221
63
49
28
14
0.1
|
1129423
99
72
36
29
0.6
|
1129441
78
45
27
12
0.2
|
1129462
95
70
64
25
0.9
|
1129482
84
63
70
29
1.0
|
1129483
69
33
14
8
0.1
|
1129501
95
66
35
14
0.4
|
1129502
93
73
49
19
0.6
|
1129542
66
34
24
12
0.1
|
1129543
89
44
30
17
0.3
|
|
TABLE 97
|
|
Dose-dependent reduction of human FXII RNA by
|
modified oligonucleotides
|
FXII (% UTC)
|
Compound
47
188
750
3000
|
Number
nM
nM
nM
nM
IC50 μM
|
|
1128763
80
57
37
22
0.3
|
1128764
73
42
26
12
0.2
|
1128802
68
58
32
19
0.2
|
1129044
88
69
43
26
0.6
|
1129065
85
60
36
15
0.4
|
1129084
87
70
47
23
0.6
|
1129104
103
87
54
40
1.4
|
1129143
81
65
40
24
0.4
|
1129144
98
68
45
30
0.7
|
1129424
95
59
39
24
0.5
|
1129445
83
54
40
20
0.4
|
1129484
61
40
22
15
0.1
|
1129485
55
39
21
10
0.1
|
1129486
57
38
21
9
0.1
|
1129503
114
110
86
43
>3.0
|
1129504
100
71
43
26
0.7
|
1129544
82
46
33
18
0.3
|
1129545
70
54
32
15
0.2
|
1129864
89
66
53
31
0.8
|
|
TABLE 98
|
|
Dose-dependent reduction of human FXII RNA
|
by modified oligonucleotides
|
FXII (% UTC)
|
Compound
47
188
750
3000
|
Number
nM
nM
nM
nM
IC50 μM
|
|
1128747
99
95
60
38
1.6
|
1128765
76
48
29
11
0.2
|
1128766
49
52
36
17
0.1
|
1128768
65
53
34
25
0.2
|
1128802
83
55
41
15
0.3
|
1128808
98
85
67
31
1.4
|
1128848
79
59
47
18
0.4
|
1129227
81
60
39
31
0.5
|
1129287
72
72
34
18
0.3
|
1129407
59
51
44
27
0.2
|
1129408
61
58
40
25
0.2
|
1129426
81
68
46
19
0.5
|
1129466
84
55
48
29
0.5
|
1129487
71
56
23
35
0.3
|
1129488
59
49
26
11
0.1
|
1129489
58
42
16
15
0.1
|
1129526
96
79
64
33
1.3
|
1129527
69
58
31
19
0.2
|
1129948
84
65
45
27
0.5
|
|
TABLE 99
|
|
Dose-dependent reduction of human FXII RNA
|
by modified oligonucleotides
|
FXII (% UTC)
|
Compound
47
188
750
3000
|
Number
nM
nM
nM
nM
IC50 μM
|
|
1128769
49
37
24
19
<0.04
|
1128802
67
49
25
12
0.2
|
1128810
75
77
43
29
0.6
|
1128830
117
72
33
29
0.7
|
1128870
49
36
38
18
<0.04
|
1129051
88
71
52
33
0.9
|
1129092
86
66
43
27
0.6
|
1129211
76
54
57
19
0.4
|
1129409
60
32
34
13
0.1
|
1129472
70
35
42
18
0.2
|
1129490
58
29
13
6
0.1
|
1129491
52
31
22
7
<0.04
|
1129492
62
26
10
9
0.1
|
1129510
55
42
42
14
0.1
|
1129529
73
50
44
21
0.3
|
1129530
66
34
24
9
0.1
|
1129531
48
42
32
10
<0.04
|
1129532
55
27
13
8
<0.04
|
1129949
84
98
56
42
2.0
|
|
TABLE 100
|
|
Dose-dependent reduction of human FXII RNA
|
by modified oligonucleotides
|
FXII (% UTC)
|
Compound
78
313
1250
5000
|
Number
nM
nM
nM
nM
IC50 μM
|
|
1130826
88
63
43
14
0.7
|
1130828
73
56
37
23
0.5
|
1130871
65
28
19
18
0.1
|
1130883
94
57
44
21
0.8
|
1131023
89
64
42
22
0.8
|
1131024
92
68
43
31
1.1
|
1131247
107
105
54
34
2.3
|
1131416
81
67
39
23
0.7
|
1131640
97
64
45
20
0.9
|
1131695
78
54
33
21
0.5
|
1131696
100
45
34
14
0.6
|
1131697
70
46
31
7
0.3
|
1131724
96
60
36
37
1.0
|
1131725
83
64
45
17
0.7
|
1131751
88
54
34
15
0.5
|
1131752
75
49
40
21
0.5
|
1131753
61
38
20
8
0.1
|
1131891
80
46
37
13
0.4
|
1131892
77
84
61
33
2.1
|
|
TABLE 101
|
|
Dose-dependent reduction of human FXII RNA
|
by modified oligonucleotides
|
FXII (% UTC)
|
Compound
78
313
1250
5000
|
Number
nM
nM
nM
nM
IC50 μM
|
|
1130829
78
62
28
18
0.5
|
1130830
92
49
26
25
0.6
|
1130871
75
53
27
15
0.4
|
1130914
100
86
73
43
4.2
|
1130970
88
84
63
39
3.0
|
1130997
87
73
57
24
1.2
|
1130998
92
76
61
45
3.4
|
1131025
71
76
42
20
0.7
|
1131053
84
85
55
32
1.8
|
1131473
90
74
35
19
0.8
|
1131474
94
77
55
36
1.9
|
1131641
82
69
43
23
0.8
|
1131671
61
49
35
21
0.3
|
1131698
88
87
48
14
1.0
|
1131699
73
43
24
11
0.3
|
1131754
48
22
14
7
<0.1
|
1131755
61
41
18
5
0.2
|
1132453
96
74
55
23
1.2
|
1132537
88
65
46
22
0.9
|
|
TABLE 102
|
|
Dose-dependent reduction of human FXII RNA
|
by modified oligonucleotides
|
FXII (% UTC)
|
Compound
78
313
1250
5000
IC50
|
Number
nM
nM
nM
nM
μM
|
|
1130831
68
50
20
11
0.3
|
1130832
77
55
35
21
0.5
|
1130858
80
88
55
35
2.0
|
1130871
72
37
24
16
0.2
|
1130973
85
61
45
22
0.8
|
1131084
85
95
49
27
1.5
|
1131198
85
68
38
24
0.8
|
1131223
91
53
41
16
0.6
|
1131309
87
64
34
42
1.1
|
1131310
86
62
51
28
1.1
|
1131700
67
45
29
9
0.2
|
1131701
55
48
26
5
0.2
|
1131702
96
42
16
9
0.4
|
1131730
93
64
38
32
1.0
|
1131756
39
13
13
5
0.1
|
1131757
54
27
14
10
0.1
|
1131758
73
58
25
9
0.4
|
1132455
88
74
59
33
1.8
|
1132651
76
76
55
38
2.0
|
|
TABLE 103
|
|
Dose-dependent reduction of human FXII RNA
|
by modified oligonucleotides
|
FXII (% UTC)
|
Compound
78
313
1250
5000
|
Number
nM
nM
nM
nM
IC50 μM
|
|
1130871
65
52
30
22
0.3
|
1130894
72
66
50
38
1.3
|
1131006
100
92
68
47
4.8
|
1131146
84
91
56
37
2.4
|
1131199
76
75
54
22
1.0
|
1131255
83
65
53
19
0.9
|
1131314
77
71
61
34
1.9
|
1131337
70
81
57
36
2.1
|
1131423
74
64
40
21
0.6
|
1131426
98
87
81
44
>5.0
|
1131481
91
66
43
28
1.0
|
1131593
66
51
37
26
0.4
|
1131673
93
63
45
35
1.2
|
1131704
96
45
39
19
0.6
|
1131706
93
80
62
40
2.7
|
1131759
62
67
40
23
0.5
|
1131760
87
71
55
40
2.0
|
1131761
89
75
60
29
1.6
|
1132797
100
75
77
54
>5.0
|
|
TABLE 104
|
|
Dose-dependent reduction of human FXII RNA
|
by modified oligonucleotides
|
FXII (% UTC)
|
Compound
78
313
1250
5000
|
Number
nM
nM
nM
nM
IC50 μM
|
|
1130839
141
86
58
32
2.0
|
1130871
57
34
22
9
0.1
|
1130920
89
83
76
23
2.1
|
1130952
94
71
49
29
1.3
|
1130979
81
84
68
34
2.7
|
1131007
104
73
61
43
2.6
|
1131120
108
68
44
42
1.6
|
1131204
71
50
27
26
0.4
|
1131340
92
78
63
27
1.7
|
1131344
88
69
44
35
1.2
|
1131368
87
79
63
41
3.0
|
1131424
74
79
47
33
1.3
|
1131428
99
64
47
21
0.9
|
1131480
82
60
57
17
0.8
|
1131484
85
67
46
24
0.9
|
1131595
67
48
35
23
0.3
|
1131736
79
78
40
14
0.7
|
1131763
49
38
22
15
<0.1
|
1131764
59
44
25
15
0.2
|
|
TABLE 105
|
|
Dose-dependent reduction of human FXII RNA
|
by modified oligonucleotides
|
FXII (% UTC)
|
Compound
78
313
1250
5000
|
Number
nM
nM
nM
nM
IC50 μM
|
|
1130840
76
69
46
27
0.9
|
1130868
54
49
33
13
0.2
|
1130870
76
57
48
18
0.6
|
1130871
72
41
29
18
0.3
|
1130925
70
60
31
19
0.4
|
1130981
86
62
42
23
0.8
|
1130983
88
62
43
29
0.9
|
1131009
92
91
50
35
1.9
|
1131036
83
72
59
32
1.7
|
1131095
74
59
31
34
0.6
|
1131375
91
89
67
46
4.9
|
1131429
75
61
45
24
0.7
|
1131624
72
56
49
22
0.6
|
1131625
61
72
43
22
0.6
|
1131627
91
58
42
20
0.7
|
1131711
94
100
53
30
1.9
|
1131737
79
62
42
19
0.6
|
1131739
94
80
58
31
1.8
|
1131767
82
50
42
34
0.8
|
|
TABLE 106
|
|
Dose-dependent reduction of human FXII RNA
|
by modified oligonucleotides
|
FXII (% UTC)
|
Compound
78
313
1250
5000
|
Number
nM
nM
nM
nM
IC50 μM
|
|
1130871
59
39
23
11
0.1
|
1130872
77
59
31
38
0.7
|
1130900
57
50
51
23
0.3
|
1130928
76
53
57
55
>5.0
|
1131013
75
58
25
16
0.4
|
1131040
61
57
43
23
0.4
|
1131096
82
70
39
35
1.1
|
1131208
102
80
57
33
1.9
|
1131347
86
67
63
31
1.7
|
1131348
74
67
42
29
0.8
|
1131432
61
45
34
22
0.2
|
1131433
91
63
28
20
0.6
|
1131599
70
61
37
23
0.5
|
1131601
88
58
49
34
1.1
|
1131630
93
54
42
20
0.7
|
1131712
73
54
32
22
0.4
|
1131713
101
66
33
27
0.9
|
1132188
89
75
69
41
3.4
|
1132271
80
76
68
48
>5.0
|
|
TABLE 107
|
|
Dose-dependent reduction of human FXII RNA
|
by modified oligonucleotides
|
FXII (% UTC)
|
Compound
78
313
1250
5000
|
Number
nM
nM
nM
nM
IC50 μM
|
|
1130763
91
71
50
44
2.1
|
1130846
64
56
42
37
0.6
|
1130871
63
41
19
16
0.2
|
1130874
69
74
40
24
0.7
|
1130903
119
90
65
35
2.4
|
1130987
66
39
27
17
0.2
|
1131015
80
84
65
33
2.4
|
1131126
66
77
51
28
1.1
|
1131155
87
79
61
31
1.9
|
1131350
79
66
46
30
1.0
|
1131434
83
56
40
23
0.6
|
1131462
73
73
56
34
1.6
|
1131463
98
104
77
45
>5.0
|
1131686
71
53
48
29
0.7
|
1131687
78
60
42
29
0.8
|
1131688
74
57
34
23
0.5
|
1131716
100
58
43
23
0.9
|
1131744
78
67
32
58
2.2
|
1132387
104
96
77
51
>5.0
|
|
TABLE 108
|
|
Dose-dependent reduction of
|
human FXII RNA by modified oligonucleotides
|
FXII (% UTC)
|
Compound
78
313
1250
5000
IC50
|
Number
nM
nM
nM
nM
μM
|
|
1130871
69.8
45.8
26.5
23.1
0.3
|
1130988
76.2
74.4
49.9
24.9
1.0
|
1131018
92.2
62.8
43.5
20.2
0.8
|
1131019
90.5
78.4
36.3
20.4
0.9
|
1131100
90.3
69.0
47.3
29.5
1.2
|
1131242
85.1
59.3
45.8
19.2
0.7
|
1131243
84.3
70.1
49.8
23.1
1.0
|
1131409
85.6
82.4
46.3
26.1
1.2
|
1131411
95.8
87.5
48.1
39.4
2.0
|
1131605
77.7
80.7
44.5
25.8
1.0
|
1131635
80.9
50.2
44.3
28.5
0.7
|
1131663
80.0
74.7
44.0
17.3
0.8
|
1131689
92.7
72.3
64.7
27.1
1.6
|
1131691
76.5
55.0
35.1
16.4
0.5
|
1131717
85.2
58.3
39.3
23.6
0.7
|
1131718
94.4
63.5
47.4
33.6
1.2
|
1131745
73.3
49.4
32.1
25.7
0.4
|
1131746
81.7
54.2
39.0
15.6
0.5
|
1131747
69.9
48.1
33.6
18.1
0.3
|
|
TABLE 109
|
|
Dose-dependent reduction of human
|
FXII RNA by modified oligonucleotides
|
FXII (% UTC)
|
Compound
78
313
1250
5000
IC50
|
Number
nM
nM
nM
nM
μM
|
|
1130824
86
73
56
29
1.4
|
1130825
111
78
49
42
2.0
|
1130871
76
39
26
17
0.3
|
1130908
76
65
51
28
1.0
|
1130938
97
77
57
19
1.2
|
1131020
79
62
47
15
0.6
|
1131021
77
72
51
27
1.1
|
1131216
78
87
56
55
>5.0
|
1131357
90
84
64
38
2.8
|
1131608
100
70
51
30
1.4
|
1131692
70
63
33
22
0.5
|
1131693
63
41
29
18
0.2
|
1131694
79
57
21
16
0.4
|
1131720
92
63
72
27
1.7
|
1131721
92
85
67
40
3.3
|
1131722
106
95
56
40
2.6
|
1131748
75
57
53
20
0.7
|
1131749
90
66
47
25
1.0
|
1132505
85
78
50
36
1.7
|
|
TABLE 110
|
|
Dose-dependent reduction of human
|
FXII RNA by modified oligonucleotides
|
FXII (% UTC)
|
Compound
78
313
1250
5000
IC50
|
Number
nM
nM
nM
nM
μM
|
|
413643
71
57
28
26
0.4
|
462073
94
68
30
23
0.8
|
462192
64
50
41
16
0.3
|
626671
101
98
74
45
>5.0
|
1124324
97
87
102
52
>5.0
|
1124408
103
74
50
22
1.2
|
1124489
86
84
75
56
>5.0
|
1124490
98
83
58
44
3.0
|
1124620
91
74
55
33
1.6
|
1124621
93
74
45
21
1.0
|
1124677
112
83
66
48
3.8
|
1124813
102
86
55
39
2.3
|
1124996
93
62
49
32
1.2
|
1125047
72
63
59
24
1.0
|
1125074
92
70
56
39
2.0
|
1125075
100
94
58
51
>5.0
|
1125101
120
108
50
43
2.7
|
1125102
105
61
57
45
2.3
|
1125570
115
113
115
81
>5.0
|
|
TABLE 111
|
|
Dose-dependent reduction of
|
human FXII RNA by modified oligonucleotides
|
FXII (% UTC)
|
Compound
78
313
1250
5000
IC50
|
Number
nM
nM
nM
nM
μM
|
|
413644
129
90
69
36
2.6
|
462073
117
78
46
36
1.6
|
1124222
490
74
81
51
>5.0
|
1124223
93
84
66
51
>5.0
|
1124328
129
93
82
42
4.0
|
1124384
80
101
79
59
>5.0
|
1124491
76
78
52
35
1.6
|
1124598
100
87
91
51
>5.0
|
1124842
105
93
65
56
>5.0
|
1124998
122
77
67
41
2.7
|
1125023
96
91
53
36
2.1
|
1125049
105
91
56
43
2.7
|
1125050
100
78
67
29
1.9
|
1125051
80
58
39
19
0.6
|
1125076
103
96
83
51
>5.0
|
1125077
106
99
74
42
4.4
|
1125078
94
92
87
40
>5.0
|
1125103
99
116
65
37
3.5
|
1125104
115
94
49
55
3.5
|
|
TABLE 112
|
|
Dose-dependent reduction of
|
human FXII RNA by modified oligonucleotides
|
FXII (% UTC)
|
Compound
250
1000
4000
IC50
|
Number
63 nM
nM
nM
nM
μM
|
|
1129531
78
53
26
11
0.3
|
1206486
82
65
51
31
0.9
|
1206977
92
69
50
23
0.8
|
1206981
79
66
45
25
0.7
|
1207005
67
41
17
13
0.2
|
1207007
51
31
15
6
<0.1
|
1207009
90
65
38
26
0.7
|
1207111
87
56
44
22
0.6
|
1207113
92
73
45
26
0.9
|
1207133
72
48
29
18
0.3
|
1207164
77
76
52
36
1.3
|
1207228
87
65
53
37
1.2
|
1207230
106
77
63
60
>4.0
|
1207244
91
57
46
24
0.7
|
1207276
63
39
11
11
0.1
|
1207362
75
71
48
20
0.6
|
1207366
94
53
38
24
0.6
|
1207370
68
55
39
21
0.3
|
1207394
61
39
22
11
0.1
|
|
TABLE 113
|
|
Dose-dependent reduction of
|
human FXII RNA by modified oligonucleotides
|
FXII (% UTC)
|
Compound
63
250
1000
4000
IC50
|
Number
nM
nM
nM
nM
μM
|
|
1129531
92
47
28
14
0.4
|
1206487
82
68
54
29
1
|
1206497
99
70
58
31
1.3
|
1206970
77
54
35
12
0.4
|
1206978
93
63
35
15
0.5
|
1206986
91
71
52
36
1.3
|
1207010
76
51
17
7
0.2
|
1207106
84
65
49
19
0.7
|
1207125
91
52
28
8
0.4
|
1207159
71
36
14
6
0.2
|
1207190
78
61
59
31
1
|
1207247
101
70
28
16
0.6
|
1207254
71
49
29
18
0.3
|
1207255
71
49
28
21
0.3
|
1207271
113
80
55
29
1.3
|
1207278
46
36
11
4
<0.1
|
1207279
66
38
23
10
0.2
|
1207358
78
61
43
17
0.5
|
1207367
79
48
30
9
0.3
|
|
TABLE 114
|
|
Dose-dependent reduction of
|
human FXII RNA by modified oligonucleotides
|
FXII (% UTC)
|
Compound
250
1000
4000
IC50
|
Number
63 nM
nM
nM
nM
μM
|
|
1129531
57
40
17
7
0.1
|
1206490
72
42
35
24
0.3
|
1206492
73
60
46
22
0.5
|
1206980
68
51
34
13
0.3
|
1206996
76
54
30
20
0.4
|
1207004
66
57
41
30
0.4
|
1207006
47
19
10
7
<0.1
|
1207118
73
53
31
14
0.3
|
1213120
60
45
24
15
0.1
|
1213135
70
53
31
23
0.3
|
1213144
42
20
25
6
<0.1
|
1213220
58
53
41
33
0.3
|
1213227
78
72
40
27
0.7
|
1213234
80
64
44
25
0.6
|
1213235
66
59
30
18
0.3
|
1213259
52
29
16
9
<0.1
|
1213260
51
35
13
13
<0.1
|
1213262
56
27
18
9
<0.1
|
1213264
77
46
18
9
0.2
|
|
TABLE 115
|
|
Dose-dependent reduction of human FXII RNA by modified oligonucleotides
|
FXII (% UTC)
|
Compound
250
1000
4000
IC50
|
Number
63 nM
nM
nM
nM
μM
|
|
1129531
77
44
20
20
0.2
|
1206489
83
53
35
21
0.4
|
1206509
97
89
72
33
2.3
|
1206971
84
65
38
22
0.6
|
1206972
69
70
49
21
0.6
|
1206984
64
51
37
25
0.3
|
1206987
70
82
64
47
>4.0
|
1207008
58
37
18
14
0.1
|
1207123
96
73
59
26
1.1
|
1207131
79
57
45
27
0.6
|
1207139
86
66
54
24
0.9
|
1207160
53
27
13
11
<0.1
|
1207161
53
31
16
13
<0.1
|
1207248
83
58
43
31
0.7
|
1207277
42
28
12
6
<0.1
|
1207312
85
75
39
23
0.7
|
1207360
80
48
39
22
0.4
|
1207381
79
85
61
46
3.6
|
1207393
58
38
17
7
0.1
|
|
TABLE 116
|
|
Dose-dependent reduction of human FXII RNA by modified oligonucleotides
|
FXII (% UTC)
|
Compound
250
1000
4000
IC50
|
Number
63 nM
nM
nM
nM
μM
|
|
1128802
88
57
36
17
0.5
|
1129531
68
44
30
14
0.2
|
1208476
71
62
35
23
0.4
|
1208506
80
80
77
67
>4.0
|
1208689
84
76
66
76
>4.0
|
1208937
92
85
83
69
>4.0
|
1209017
80
73
72
59
>4.0
|
1209119
80
66
72
59
>4.0
|
1209422
94
90
54
61
>4.0
|
1209543
91
94
83
76
>4.0
|
1209545
88
82
71
70
>4.0
|
1209565
98
95
70
78
>4.0
|
1209576
82
83
68
60
>4.0
|
1209607
93
88
74
85
>4.0
|
1209615
97
83
68
71
>4.0
|
1209616
82
76
70
60
>4.0
|
1209630
92
86
83
71
>4.0
|
1209659
91
98
67
57
>4.0
|
1209674
99
81
81
70
>4.0
|
|
TABLE 117
|
|
Dose-dependent reduction of
|
human FXII RNA by modified oligonucleotides
|
FXII (% UTC)
|
Compound
250
1000
4000
IC50
|
Number
63 nM
nM
nM
nM
μM
|
|
1128802
82
55
38
16
0.4
|
1129531
77
38
29
9
0.2
|
1208612
83
79
63
58
>4.0
|
1208757
84
76
74
69
>4.0
|
1208977
76
75
76
43
>4.0
|
1209134
81
78
75
72
>4.0
|
1209238
76
73
68
71
>4.0
|
1209391
91
100
89
78
>4.0
|
1209430
84
77
73
54
>4.0
|
1209455
77
74
71
71
>4.0
|
1209514
76
77
70
73
>4.0
|
1209547
89
78
58
35
1.6
|
1209548
81
74
59
35
1.5
|
1209605
103
100
78
80
>4.0
|
1209624
89
90
75
70
>4.0
|
1209645
105
85
79
62
>4.0
|
1209670
115
96
82
85
>4.0
|
1209688
78
75
80
76
>4.0
|
1209692
88
97
89
71
>4.0
|
|
Example 8: Design of Oligomeric Compounds Complementary to a Human FXII Nucleic Acid
Oligomeric compounds were designed as indicated in the tables below. Modified oligonucleotides described in the Examples above (parent compounds) were further modified by either adding a THA-C6-GalNAc3 conjugate (designated as [THA-GalNAc] in the table below) at the 5′ end of the modified oligonucleotide, or by adding a 3′-THA-C6-GalNAc hydroxyproline PO conjugate (designated as 3′-HPPO-GalNAc in the tables below), at the 3′-end of the modified oligonucleotide. THA-GalNAc is represented by the structure below wherein the phosphate group is attached to the 5′-oxygen atom of the 5′ nucleoside:
![embedded image]()
3′-HPPO-GalNAc is represented by the structure below wherein the phosphate group is attached to the 3′-oxygen atom of the 3′ nucleoside:
![embedded image]()
The chemistry notation column in the tables below specifies the specific chemistry notation for modified oligonucleotides; wherein subscript ‘d’ represents a 2′-β-D-deoxyribosyl sugar moiety, subscript ‘e’ represents a 2′-MOE sugar moiety, subscript ‘y’ represents a 2′-O-methyl sugar moiety, subscript ‘k’ represents a cEt modified sugar moiety, subscript ‘s’ represents a phosphorothioate internucleoside linkage, subscript ‘o’ represents a phosphodiester internucleoside linkage, and superscript ‘m’ before the cytosine residue (mC) represents a 5-methyl cytosine.
TABLE 118
|
|
Design of GalNAc conjugated modified oligonucleotides complementary to human FXII
|
Parent
|
Compound
Compound
SEQ ID
|
No.
No.
Sequence and Chemistry notation (5′ to 3′)
No.
|
|
1194343
1128769
THA-GalNAc-oAksGksAksAdsmCdsGdsAdsmCdsTdsGdsTdsGdsTdsGksmCksTk
3723
|
|
1194345
1128801
THA-GalNAc-oAksmCksAksTdsTdsTdsGdsTdsGdsGdsTdsAdsmCdsAksGksmCk
3114
|
|
1194346
1128802
THA-GalNAc-oTksAksmCksAdsTdsTdsTdsGdsTdsGdsGdsTdsAdsmCksAksGk
2426
|
|
1194347
1128870
THA-GalNAc-oTksGksTksGdsGdsAdsAdsAdsAdsAdsmCdsmCdsGdsGksAksGk
3728
|
|
1194348
1129221
THA-GalNAc-omCksGksTksmCdsTdsGdsGdsmCdsAdsmCdsGdsGdsmCdsTksmCksAk
3060
|
|
1194349
1129287
THA-GalNAc-omCksGksTksAdsAdsGdsGdsmCdsGdsAdsmCdsAdsGdsGksAksGk
3521
|
|
1194350
1129407
THA-GalNAc-oTksGksmCksmCdsTdsTdsGdsmCdsAdsGdsGdsGdsTdsGksAksGk
3527
|
|
1194357
1129485
THA-GalNAc-oGksmCksGksGdsAdsAdsTdsmCdsAdsmCdsmCdsAdsAdsGksGksAk
3379
|
|
1194358
1129486
THA-GalNAc-oTksGksmCksGdsGdsAdsAdsTdsmCdsAdsmCdsmCdsAdsAksGksGk
3455
|
|
1194359
1129487
THA-GalNAc-omCksTksGksmCdsGdsGdsAdsAdsTdsmCdsAdsmCdsmCdsAksAksGk
3531
|
|
1194360
1129488
THA-GalNAc-oAksmCksTksGdsmCdsGdsGdsAdsAdsTdsmCdsAdsmCdsmCksAksAk
3607
|
|
1194361
1129489
THA-GalNAc-omCksAksmCksTdsGdsmCdsGdsGdsAdsAdsTdsmCdsAdsmCksmCksAk
3683
|
|
1194363
1129492
THA-GalNAc-oTksmCksTksmCdsAdsmCdsTdsGdsmCdsGdsGdsAdsAdsTksmCksAk
3911
|
|
1194370
1129534
THA-GalNAc-oAksTksTksGdsAdsGdsTdsTdsmCdsmCdsTdsGdsmCdsGksmCksmCk
2540
|
|
1194371
1129538
THA-GalNAc-oAksmCksTksTdsTdsAdsTdsTdsGdsAdsGdsTdsTdsmCksmCksTk
2848
|
|
1194372
1129540
THA-GalNAc-oGksmCksAksmCdsTdsTdsTdsAdsTdsTdsGdsAdsGdsTksTksmCk
3000
|
|
1194373
1129542
THA-GalNAc-oAksAksGksmCdsAdsmCdsTdsTdsTdsAdsTdsTdsGdsAksGksTk
3147
|
|
1194374
1129543
THA-GalNAc-oAksAksAksGdsmCdsAdsmCdsTdsTdsTdsAdsTdsTdsGksAksGk
3229
|
|
1194375
1129544
THA-GalNAc-omCksAksAksAdsGdsmCdsAdsmCdsTdsTdsTdsAdsTdsTksGksAk
3306
|
|
1194376
1129545
THA-GalNAc-oTksmCksAksAdsAdsGdsmCdsAdsmCdsTdsTdsTdsAdsTksTksGk
3382
|
|
1194377
1129947
THA-GalNAc-oTksGksmCksTdsTdsTdsmCdsmCdsGdsmCdsAdsmCdsTdsmCksTksmCk
3554
|
|
1194378
1130831
THA-GalNAc-oGesAeoAeomCeoGeoAdsmCdsTdsGdsTdsGdsTdsGdsmCdsTdsmCeoTeoTes
997
|
mCesAe
|
|
1194379
1130868
THA-GalNAc-oAesmCeoAeoTeoTeoTdsGdsTdsGdsGdsTdsAdsmCdsAdsGdsmCeoTeoGesm
1756
|
CesmCe
|
|
1194380
1130871
THA-GalNAc-oGesGeoTeoAeomCeoAdsTdsTdsTdsGdsTdsGdsGdsTdsAdsmCeoAeoGesm
183
|
CesTe
|
|
1194381
1130900
THA-GalNAc-oAesTeomCeoGeomCeoTdsGdsGdsTdsmCdsmCdsTdsGdsAdsTdsmCeoAeo
1906
|
AesAesGe
|
|
1194382
1130925
THA-GalNAc-oGesGeoTeomCeoTeoTdsTdsmCdsAdsmCdsTdsTdsTdsmCdsTdsTeoGeoGes
1758
|
GesmCe
|
|
1194383
1130987
THA-GalNAc-oGesGeomCeoTeomCeoAdsAdsAdsGdsmCdsAdsmCdsTdsTdsmCdsTeomCeo
2135
|
TesTesTe
|
|
1194384
1131013
THA-GalNAc-oAesmCeomCeoAeoTeoAdsTdsmCdsTdsmCdsAdsTdsTdsmCdsTdsTeoGeoTes
1984
|
GesGe
|
|
1194385
1131040
THA-GalNAc-oAesmCeomCeomCeoTeoTdsGdsmCdsAdsmCdsTdsGdsGdsmCdsAdsTeomCeo
1911
|
TesGesGe
|
|
1194387
1131204
THA-GalNAc-oAesGeoTeoTeomCeomCdsGdsmCdsGdsmCdsTdsTdsGdsmCdsTdsmCeoGeo
1618
|
GesmCesAe
|
|
1194388
1131432
THA-GalNAc-oGesGeoTeoAeoGeoGdsAdsGdsmCdsGdsmCdsAdsmCdsGdsGdsmCeomCeo
1925
|
AesAesmCe
|
|
1194389
1131593
THA-GalNAc-oTesGeoAeoTeoGeomCdsmCdsTdsTdsGdsmCdsAdsGdsGdsGdsTeoGeoAes
1326
|
GesmCe
|
|
1194390
1131595
THA-GalNAc-oGesAeoTeoGeoAeoTdsGdsmCdsmCdsTdsTdsGdsmCdsAdsGdsGeoGeoTes
1556
|
GesAe
|
|
1194391
1131671
THA-GalNAc-oGesmCeoAeoAeoTeomCdsAdsGdsGdsAdsAdsAdsmCdsGdsGdsTeoGeoTes
950
|
GesmCe
|
|
1194392
1131693
THA-GalNAc-oTesGeomCeoGeoGeoAdsAdsTdsmCdsAdsmCdsmCdsAdsAdsGdsGeoAeoGes
496
|
GesGe
|
|
1194393
1131694
THA-GalNAc-omCesTeoGeomCeoGeoGdsAdsAdsTdsmCdsAdsmCdsmCdsAdsAdsGeoGeo
571
|
AesGesGe
|
|
1194394
1131697
THA-GalNAc-omCesTeomCeoAeomCeoTdsGdsmCdsGdsGdsAdsAdsTdsmCdsAdsmCeomCeo
802
|
AesAesGe
|
|
1194395
1131699
THA-GalNAc-oTesmCeoTeomCeoTeomCdsAdsmCdsTdsGdsmCdsGdsGdsAdsAdsTeomCeo
951
|
AesmCesmCe
|
|
1194396
1131700
THA-GalNAc-omCesTeomCeoTeomCeoTdsmCdsAdsmCdsTdsGdsmCdsGdsGdsAdsAeoTeom
1028
|
CesAesmCe
|
|
1194397
1131701
THA-GalNAc-oAesmCeoTeomCeoTeomCdsTdsmCdsAdsmCdsTdsGdsmCdsGdsGdsAeoAeo
1179
|
TesmCesAe
|
|
1194398
1131702
THA-GalNAc-omCesmCeoAeomCeoTeomCdsTdsmCdsTdsmCdsAdsmCdsTdsGdsmCdsGeo
1103
|
GeoAesAesTe
|
|
1194399
1131712
THA-GalNAc-omCesTeoTeomCeomCeoAdsTdsGdsmCdsmCdsmCdsmCdsAdsGdsmCdsmCeo
1935
|
AeomCesTesmCe
|
|
1194400
1131745
THA-GalNAc-oAesGeoTeoTeomCeomCdsTdsGdsmCdsGdsmCdsmCdsAdsTdsmCdsmCeoTeo
2309
|
GesGesmCe
|
|
1194401
1131747
THA-GalNAc-oTesGeoAeoGeoTeoTdsmCdsmCdsTdsGdsmCdsGdsmCdsGdsmCdsmCdsAdsTeomCeom
347
|
CesTesGe
|
|
1194402
1131753
THA-GalNAc-omCesAeomCeoTeoTeoTdsAdsTdsTdsGdsAdsGdsTdsTdsmCdsmCeoTeoGesm
163
|
CesGe
|
|
1194403
1131754
THA-GalNAc-oGesmCeoAeomCeoTeoTdsTdsAdsTdsTdsGdsAdsGdsTdsTdsmCeomCeoTes
164
|
GesmCe
|
|
1194404
1131755
THA-GalNAc-oAesGeomCeoAeomCeoTdsTdsTdsAdsTdsTdsGdsAdsGdsTdsTeomCeomCes
953
|
TesGe
|
|
1194405
1131756
THA-GalNAc-oAesAeoGeomCeoAeomCdsTdsTdsTdsAdsTdsTdsGdsAdsGdsTeoTeomCesm
1030
|
CesTe
|
|
1194406
1131757
THA-GalNAc-oAesAeoAeoGeomCeoAdsmCdsTdsTdsTdsAdsTdsTdsGdsAdsGeoTeoTesm
165
|
CesmCe
|
|
1194407
1131758
THA-GalNAc-omCesAeoAeoAeoGeomCdsAdsmCdsTdsTdsTdsAdsTdsTdsGdsAeoGeoTes
1105
|
TesmCe
|
|
1194408
1131763
THA-GalNAc-oGesmCeoAeoTeoTeoTdsTdsmCdsAdsAdsAdsGdsmCdsAdsmCdsTeoTeoTes
1562
|
AesTe
|
|
1194409
1131764
THA-GalNAc-oAesGeomCeoAeoTeoTdsTdsTdsmCdsAdsAdsAdsGdsmCdsAdsmCeoTeoTes
97
|
TesAe
|
|
1194410
1131891
THA-GalNAc-omCesmCeoGeoAeomCeoTdsGdsTdsGdsTdsGdsmCdsTdsmCdsTdsTeomCeo
659
|
AesGesmCe
|
|
1194411
1132187
THA-GalNAc-oGesTeoTeoTeoGeomCdsTdsAdsGdsTdsTdsmCdsAdsAdsTdsGeoTeoTesmCes
1876
|
Ae
|
|
1194412
1132188
THA-GalNAc-oAesmCeoTeoGeoTeoTdsTdsGdsmCdsTdsAdsGdsTdsTdsmCdsAeoAeoTesGes
1951
|
Te
|
|
1194413
1132189
THA-GalNAc-oAesAeoTeoAeomCeoTdsGdsTdsTdsTdsGdsmCdsTdsAdsGdsTeoTeomCes
2025
|
AesAe
|
|
1194415
1132270
THA-GalNAc-oaesTeoTeomCeomCeomCdsAdsGdsTdsTdsAdsAdsGdsGdsTdsTomCeoAes
1730
|
AesmCe
|
|
1194416
1132271
THA-GalNAc-omCesAeomCeoAeoTeoTdsmCdsmCdsmCdsAdsGdsTdsTdsAdsAdsGeoGeo
1879
|
TesTesmCe
|
|
1194417
1132272
THA-GalNAc-oTesTeoGeomCeoAeomCdsAdsTdsTdsmCdsmCdsmCdsAdsGdsTdsTeoAeoAes
1954
|
GesGe
|
|
1194419
1132504
THA-GalNAc-oGesTeoTeoTemCeomCdsAdsAdsGdsmCdsTdsTdsGdsGdsTdsTeoTeoAesm
449
|
CesmCe
|
|
1194420
1132505
THA-GalNAc-omCesAeoAeoGeoTeoTdsTdsmCdsmCdsAdsAdsGdsmCdsTdsTdsGeoGeoTes
525
|
TesTe
|
|
1194421
1132506
THA-GalNAc-omCesTeomCeomCeoAeoAdsGdsTdsTdsTdsmCdsmCdsAdsAdsGdsmCeoTeo
600
|
TesGesGe
|
|
1194423
1132650
THA-GalNAc-oTesTeoGeomCeomCeoTdsTdsGdsGdsTdsGdsTdsmCdsTdsGdsAeoGeoGes
910
|
AesGe
|
|
1194424
1132651
THA-GalNAc-oAesGeomCeoTeoTeoGdsmCdsmCdsTdsTdsGdsGdsTdsGdsTdsmCeoTeoGes
985
|
AesGe
|
|
1194425
1132652
THA-GalNAc-oAesGeomCeoAeoGeomCdsTdsTdsGdsmCdsmCdsTdsTdsGdsGdsTeoGeoTes
1062
|
mCesTe
|
|
1194426
1132796
THA-GalNAc-oGesAeoAeomCeomCeoAdsAdsGdsAdsGdsmCdsAdsAdsGdsTdsTeomCeoGes
1523
|
AesTe
|
|
1194427
1132797
THA-GalNAc-oAesGeoGeoGeoAeodsmCdsmCdsAdsAdsGdsAdsGdsmCdsAdsAeoGeoTes
1369
|
TesmCe
|
|
1194429
1194286
THA-GalNAc-omCksTksTksTdsmCdsmCdsGdsmCdsAdsmCdsTdsmCdsTdsmCksmCksmCk
4026
|
|
1194430
1194287
THA-GalNAc-oGksmCksTksTdsTdsmCdmCdsGdsmCdsAdsmCdsTdsmCdsTksmCksmCk
4102
|
|
1194483
1129948
AksTksmCksTdsGdsmCdsTdsTdsTdsmCdsmCdsGdsmCdsAksmCksTk-3′-HPPO-GalNAc
3630
|
|
1194484
1129949
TksmCksTksmCdsAdsTdsmCdsTdsGdsmCdsTdsTdsTdsmCksmCksGk-3′-HPPO-GalNAc
3706
|
|
1194486
1128769
AksGksAksAdsmCdsGdsAdsmCdsTdsGdsTdsGdsTdsGksmCksTk-3′-HPPO-GalNAc
3723
|
|
1194487
1128796
TksGksTksGdsGdsTdsAdsmCdsAdsGdsmCdsTdsGdsmCksmCksGk-3′-HPPO-GalNAc
2734
|
|
1194488
1128801
AksmCksAksTdsTdsTdsGdsTdsGdsGdsTdsAdsmCdsAksGksmCk-3′-HPPO-GalNAc
3114
|
|
1194489
1128802
TksAksmCksAdsTdsTdsTdsGdsTdsGdsGdsTdsAdsmCksAksGk-3′-HPPO-GalNAc
2426
|
|
1194490
1128870
TksGksTksGdsGdsAdsAdsAdsAdsAdsmCdsmCdsGdsGksAksGk-3′-HPPO-GalNAc
3728
|
|
1194491
1129221
mCksGksTksmCdsTdsGdsGdsmCdsAdsmCdsGdsGdsmCdsTksmCksAk-3′-HPPO-GalNAc
3060
|
|
1194492
1129287
mCksGksTksAdsAdsGdsGdsmCdsGdsAdsmCdsAdsGdsGksAksGk-3′-HPPO-GalNAc
3521
|
|
1194493
1129407
TksGksmCksmCdsTdsTdsGdsmCdsAdsGdsGdsGdsTdsGksAksGk-3′-HPPO-GalNAc
3527
|
|
1194494
1129408
AksTksGksAdsTdsGdsmCdsmCdsTdsTdsGdsmCdsAdsGksGksGk-3′-HPPO-GalNAc
3603
|
|
1194495
1129409
mCksTksGksAdsTdsGdsAdsTdsGdsmCdsmCdsTdsTdsGksmCksAk-3′-HPPO-GalNAc
3679
|
|
1194496
1129441
AksGksTksAdsGdsGdsmCdsmCdsAdsmCdsAdsTdsmCdsGksGksTk-3′-HPPO-GalNAc
3071
|
|
1194497
1129472
TksGksAksGdsTdsmCdsmCdsmCdsTdsGdsAdsGdsmCdsAksAksTk-3′-HPPO-GalNAc
3910
|
|
1194498
1129476
AksAksGksAdsTdsGdsAdsGdsTdsmCdsmCdsmCdsTdsGksAksGk-3′-HPPO-GalNAc
2691
|
|
1194499
1129483
GksGksAksAdsTdsmCdsAdsmCdsmCdsAdsAdsGdsGdsAksGksGk-3′-HPPO-GalNAc
3226
|
|
1194500
1129485
GksmCksGksGdsAdsAdsTdsmCdsAdsmCdsmCdsAdsAdsGksGksAk-3′-HPPO-GalNAc
3379
|
|
1194501
1129486
TksGksmCksGdsGdsAdsAdsTdsmCdsAdsmCdsmCdsAdsAksGksGk-3′-HPPO-GalNAc
3455
|
|
1194502
1129487
mCksTksGksmCdsGdsGdsAdsAdsTdsmCdsAdsmCdsmCdsAksAksGk-3′-HPPO-GalNAc
3531
|
|
1194503
1129488
AksmCksTksGdsmCdsGdsGdsAdsAdsTdsmCdsAdsmCdsmCksAksAk-3′-HPPO-GalNAc
3607
|
|
1194504
1129489
mCksAksmCksTdsGdsmCdsGdsGdsAdsAdsTdsmCdsAdsmCksmCksAk-3′-HPPO-GalNAc
3683
|
|
1194505
1129491
mCksTksmCsAdsmCdsTdsGdsmCdsGdsGdsAdsAdsTdsmCksAksmCk-3′-HPPO-GalNAc
3835
|
|
1194506
1129492
TksmCksTksmCdsAdsmCdsTdsGdsmCdsGdsGdsAdsAdsTksmCksAk-3′-HPPO-GalNAc
3911
|
|
1194507
1129510
AksGksmCksTdsGdsGdsmCdsmCdsGdsmCdsAdsmCdsTdsGksGksGk-3′-HPPO-GalNAc
3760
|
|
1194508
1129527
TksmCksmCksTdsGdsmCdsGdsmCdsmCdsAdsTdsmCdsmCdsTksGksGk-3′-HPPO-GalNAc
3533
|
|
1194509
1129530
AksGksTksTdsmCdsmCdsTdsGdsmCdsGdsmCdsmCdsAdsTksmCksmCk-3′-HPPO-GalNAc
3761
|
|
1194510
1129531
GksAksGksTdsTdsmCdsmCdsTdsGdsmCdsGdsmCdsmCdsAksTksmCk-3′-HPPO-GalNAc
3837
|
|
1194512
1129533
TksTksGksAdsGdsTdsTdsmCdsmCdsTdsGdsmCdsGdsmCksmCksAk-3′-HPPO-GalNAc
2463
|
|
1194513
1129534
AksTksTksGdsAdsGdsTdsTdsmCdsmCdsTdsGdsmCdsGksmCksmCk-3′-HPPO-GalNAc
2540
|
|
1194514
1129538
AksmCksTksTdsTdsAdsTdsTdsGdsAdsGdsTdsTdsmGksmCksTk-3′-HPPO-GalNAc
2848
|
|
1194515
1129540
GksmCksAksmCdsTdsTdsTdsAdsTdsTdsGdsAdsGdsTksTksmCk-3′-HPPO-GalNAc
3000
|
|
1194516
1129542
AksAksGksmCdsAdsmCdsTdsTdsTdsAdsTdsTdsGdsAksGksTk-3′-HPPO-GalNAc
3147
|
|
1194517
1129543
AksAksAksGdsmCdsAdsmCdsTdsTdsTdsAdsTdsTdsGksAksGk-3′-HPPO-GalNAc
3229
|
|
1194518
1129544
mCksAksAksAdsGdsmCdsAdsmCdsTdsTdsTdsAdsTdsTksGksAk-3′-HPPO-GalNAc
3306
|
|
1194519
1129545
TksmCksAksAdsAdsGdsmCdsAdsmCdsTdsTdsTdsAdsTksTksGk-3′-HPPO-GalNAc
3382
|
|
1194520
1129947
TksGksmCksTdsTdsTdsmCdsmCdsGdsmCdsAdsmCdsTdsmCksTksmCk-3′-HPPO-GalNAc
3554
|
|
1194572
1194286
mCksTksTksTdsmCdsmCdsGdsmCdsAdsmCdsTdsmCdsTdsmCksmCksmCk-3′-HPPO-GalNAc
4026
|
|
1194573
1194287
GksmCksTksTdsTdsmCdsmCdsGdsmCdsAdsmCdsTdsmCdsTksmCksmCk-3′-HPPO-GalNAc
4102
|
|
1194574
1194288
mCksTksGksmCdsTdsTdsTdsmCdsmCdsGdsmCdsAdsmCdsTksmCksTk-3′-HPPO-GalNAc
4253
|
|
1194575
1194289
TksmCksTksGdsmCdsTdsTdsTdsmCdsmCdsGdsmCdsAdsmCksTksmCk-3′-HPPO-GalNAc
4329
|
|
1194576
1194290
mCksAksTksmCdsTdsGdsmCdsTdsTdsTdsmCdsmCdsGdsmCksAksmCk-3′-HPPO-GalNAc
4405
|
|
1194578
1194292
mCksTksmCksAdsTdsmCdsTdsGdsmCdsTdsTdsTdsmCdsmCksGksmCk-3′-HPPO-GalNAc
4557
|
|
1194579
1194293
mCksTksmCksTdsmCdsAdsTdsmCdsTdsGdsmCdsTdsTdsTksmCksmCk-3′-HPPO-GalNAc
4708
|
|
1194580
1194294
mCksmCksTksmCdsTdsmCdsAdsTdsmCdsTdsGdsmCdsTdsTksTksmCk-3′-HPPO-GalNAc
4783
|
|
1194581
1194295
mCksmCksmCksTdsmCdsTdsmCdsAdsTdsmCdsTdsGdsmCdsTksTksTk-3′-HPPO-GalNAc
4859
|
|
1194582
1194296
TksmCksmCksmCdsTdsmCdsTdsmCdsAdsTdsmCdsTdsGdsmCksTksTk-3′-HPPO-GalNAc
3950
|
|
1194583
1194297
mCksTksmCksmCdsmCdsTdsmCdsTdsmCdsAdsTdsmCdsTdsGksmCksTk-3′-HPPO-GalNAc
4027
|
|
1270699
1207006
THA-GalNAc-omCkAesmCksTdsTdsTdsAdsTdsTdsGdsAdsGdsTesTesmCksmCk
2924
|
|
1270700
1207007
THA-GalNAc-oGksmCesAksmCdsTdsTdsTdsAdsTdsTdsGdsAdsGesTesTksmCk
3000
|
|
1270701
1207160
THA-GalNAc-omCksAksmCdsTdsTdsTdsAdsTdsTdsGdsAdsGesTesTesmCksmCk
2924
|
|
1270702
1207161
THA-GalNAc-oGksmCksAdsmCdsTdsTdsTdsAdsTdsTdsGdsAesGesTesTksmCk
3000
|
|
1270703
1207277
THA-GalNAc-omCksAksmCdsTdsTdsTdsAdsTdsTdsGdsAdsGesTksTesmCksmCe
2924
|
|
1270705
1213144
THA-GalNAc-oGksmCksAksmCdsUyTdsTdsAdsTdsTdsGdsAdsGdsTksTksmCk
5006
|
|
1270707
1213260
THA-GalNAc-omCksAksmCksTdsTdsUysAdsTdsTdsGdsAdsGdsTdsTksmCksmCk
5035
|
|
1270710
1207393
THA-GalNAc-oAksmCksTdsTdsTdsAdsTdsTdsGdsAdsGdsTksTesmCksmCesTk
2848
|
|
1270712
1207394
THA-GalNAc-omCksAksmCdsTdsTdsTdsAdsTdsTdsGdsAdsGksTesTksmCesmCk
2924
|
|
1270714
1207279
THA-GalNAc-oAksGksmCdsAdsmCdsTdsTdsTdsAdsTdsTdsGesAksGesTksTe
3076
|
|
1270715
1207159
THA-GalNAc-oAksmCksTdsTdsTdsAdsTdsTdsGdsAdsGdsTesTesmCesmCksTk
2848
|
|
1270716
1207005
THA-GalNAc-oAksmCesTksTdsTdsAdsTdsTdsGdsAdsGdsTdsTesmCesmCksTk
2848
|
|
Example 9: Design of Oligomeric Compounds Complementary to a Human FXII Nucleic Acid
Modified oligonucleotides were designed as indicated in the tables below. The chemistry notation column in the tables below specifies the specific chemistry notation for modified oligonucleotides; wherein subscript ‘d’ represents a 2′-β-D-deoxyribosyl sugar moiety, subscript ‘e’ represents a 2′-MOE sugar moiety, subscript ‘y’ represents a 2′-O-methyl sugar moiety, subscript ‘k’ represents a cET modified sugar moiety, subscript ‘s’ represents a phosphorothioate internucleoside linkage, subscript ‘o’ represents a phosphodiester interrnucleoside linkage, and superscript ‘m’ before the cytosine residue (mC) represents a 5-methyl cytosine.
“Start site” indicates the 5′-most nucleoside of the target sequence to which the modified oligonucleotide is complementary. “Stop site” indicates the 3′-most nucleoside of the target sequence to which the modified oligonucleotide is complementary. As shown in the tables below, the modified oligonucleotides are complementary to either the human FXII mRNA, designated herein as SEQ ID NO: 1 (described herein above) or to the human FXII genomic sequence, designated herein as SEQ ID NO: 2 (described herein above) or to both. ‘N/A’ indicates that the modified oligonucleotide is not complementary to that particular target sequence with 100% complementarity.
TABLE 119
|
|
5-10-5 2′O-methyl modified oligonucleotides complementary to human FXII
|
SEQ
SEQ
SEQ
SEQ
|
ID NO:
ID NO:
ID NO:
ID NO:
|
Compound
1 Start
1 Stop
2 Start
2 Stop
Sequence and Chemistry notation
SEQ ID
|
Number
Site
Site
Site
Site
(5′ to 3′)
No.
|
|
1213034
2014
2033
7402
7421
CysAysCysUysUysTdsAdsTdsTdsGdsAdsGds
5039
|
TdsTdsmCdsCysUysGysCysGy
|
|
1358035
1936
1955
7324
7343
CysCysCysCysAysGdsmCdsmCdsAdsmCdsTds
5040
|
mCdsTdsmCdsTdsCysAysCysUysGy
|
|
1213030
1924
1943
7312
7331
UysCysUysCysAysmCdsTdsGdsmCdsGdsGds
5041
|
AdsAdsTdsmCdsAysCysCysAysAy
|
|
1213029
1914
1933
7302
7321
GysAysAysUysCysAdsmCdsmCdsAdsAdsGds
5042
|
GdsAdsGdsGdsGysAysAysAysGy
|
|
Example 10: Tolerability of Modified Oligonucleotides Targeting Human FXII in CD-1 Mice
CD-1 mice are a multipurpose mouse model frequently utilized for safety and efficacy testing. The mice were treated with modified oligonucleotides selected from studies described above and evaluated for changes in the levels of various plasma chemistry markers.
Study 1
Groups of four 7-to-8-week-old male CD-1 mice were injected subcutaneously once a week for six weeks (for a total of 6 treatments) with 80 mg/kg of modified oligonucleotides. One group of four male CD-1 mice was injected with saline. Mice were euthanized 72 hours following the final administration.
To evaluate the effect of modified oligonucleotides on liver and kidney function, plasma levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TBIL), blood urea nitrogen (BUN), creatinine (CRT) and albumin were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400c, Melville, NY). The results are presented in the table below. Modified oligonucleotides that caused changes in the levels of any of the liver or kidney function markers outside the expected range for modified oligonucleotides were excluded in further studies.
TABLE 120
|
|
Plasma chemistry markers in male CD-1 mice
|
Compound
AST
ALT
TBIL
BUN
CRT
Albumin
|
No.
(IU/L)
(IU/L)
(mg/dL)
(mg/dL)
(mg/dL)
(mg/dL)
|
|
saline
51
33
0.27
25
0.05
2.82
|
1194378
603
1124
0.34
27
0.07
2.61
|
1194379
255
740
0.25
22
0.04
2.65
|
1194380
62
58
0.17
26
0.08
2.84
|
1194381
368
375
0.18
24
0.09
2.83
|
1194382
380
423
0.21
25
0.08
2.67
|
1194383
72
104
0.15
25
0.06
2.54
|
1194384
66
41
0.17
23
0.05
2.70
|
1194385
1287
1501
0.24
27
0.08
2.10
|
1194387
157
239
0.26
27
0.06
2.69
|
1194388
68
48
0.17
24
0.07
2.77
|
1194389
69
53
0.16
25
0.06
2.75
|
1194390
870
1395
0.22
26
0.08
2.84
|
1194391
49
40
0.17
23
0.07
2.81
|
1194392
62
56
0.16
27
0.07
2.75
|
1194393
51
39
0.16
22
0.07
2.58
|
1194394
99
106
0.20
24
0.07
2.84
|
1194395
71
112
0.13
24
0.05
2.53
|
1194396
1614
1954
1.81
18
0.03
2.54
|
1194397
678
836
0.20
17
0.06
2.62
|
1194398
713
867
0.20
23
0.07
2.76
|
1194399
548
393
0.26
26
0.07
2.84
|
1194400
854
1165
0.66
22
0.07
2.78
|
1194401
457
1201
0.23
27
0.05
2.76
|
|
Body weights of CD-1 mice were measured at days 1 and 40, and the average body weight for each group is presented in the table below. Liver, kidney and spleen weights were measured at the end of the study and are presented in the table below. Modified oligonucleotides that caused any changes in organ weights outside the expected range for modified oligonucleotides were excluded from further studies.
TABLE 121
|
|
Body and organ weights (in grams)
|
Compound
Body Weight (g)
Liver
Kidney
Spleen
|
No.
Day 1
Day 40
(g)
(g)
(g)
|
|
saline
31
36
2.0
0.6
0.1
|
1194378
32
39
2.6
0.6
0.1
|
1194379
32
38
2.7
0.6
0.1
|
1194380
32
38
2.6
0.6
0.1
|
1194381
32
40
2.8
0.6
0.1
|
1194382
32
38
2.5
0.6
0.1
|
1194383
31
36
2.2
0.5
0.1
|
1194384
32
40
2.2
0.6
0.1
|
1194385
31
39
3.0
0.6
0.1
|
1194387
30
37
2.2
0.5
0.3
|
1194388
31
38
2.5
0.6
0.2
|
1194389
30
37
2.3
0.6
0.1
|
1194390
32
38
3.2
0.6
0.1
|
1194391
30
37
2.4
0.6
0.1
|
1194392
32
38
2.2
0.6
0.1
|
1194393
31
38
2.0
0.6
0.1
|
1194394
31
37
2.1
0.6
0.1
|
1194395
32
37
2.4
0.6
0.1
|
1194396
31
35
2.0
0.6
0.1
|
1194397
32
40
2.8
0.6
0.1
|
1194398
32
38
3.0
0.6
0.2
|
1194399
32
38
2.9
0.6
0.2
|
1194400
31
38
3.2
0.6
0.1
|
1194401
33
41
2.6
0.6
0.2
|
|
Study 2
Groups of four 7-to-8-week-old male CD-1 mice were injected subcutaneously once a week for six weeks (for a total of 6 treatments) with 80 mg/kg of modified oligonucleotides. One group of four male CD-1 mice was injected with saline. Mice were euthanized 72 hours following the final administration.
To evaluate the effect of modified oligonucleotides on liver and kidney function, plasma levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TBIL), blood urea nitrogen (BUN), creatinine (CRT) and albumin were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400c, Melville, NY). The results are presented in the table below. Modified oligonucleotides that caused changes in the levels of any of the liver or kidney function markers outside the expected range for modified oligonucleotides were excluded in further studies.
TABLE 122
|
|
Plasma chemistry markers in male CD-1 mice
|
Compound
AST
ALT
TBIL
BUN
CRT
Albumin
|
No.
(IU/L)
(IU/L)
(mg/dL)
(mg/dL)
(mg/dL)
(mg/dL)
|
|
saline
70
68
0.16
22
0.09
2.84
|
1194402
66
66
0.17
24
0.09
2.73
|
1194403
67
64
0.15
26
0.09
2.64
|
1194404
77
82
0.17
25
0.10
2.91
|
1194405
100
155
0.14
25
0.07
2.53
|
1194406
95
84
0.15
25
0.07
2.60
|
1194407
94
69
0.11
19
0.07
2.56
|
1194408
111
137
0.15
26
0.06
2.54
|
1194409
134
102
0.16
25
0.06
2.69
|
1194410
344
385
0.18
28
0.08
2.53
|
1194411
321
242
0.15
22
0.08
2.53
|
1194412
227
333
0.17
25
0.06
2.49
|
1194413
110
61
0.16
24
0.10
2.79
|
1194415
143
132
0.13
25
0.08
2.39
|
1194416
73
107
0.11
21
0.06
2.43
|
1194417
488
672
0.24
23
0.05
2.43
|
1194419
72
60
0.14
28
0.08
2.64
|
1194420
104
86
0.15
26
0.09
2.72
|
1194421
548
502
0.26
26
0.12
2.99
|
1194423
255
443
0.11
25
0.04
1.54
|
1194424
325
423
0.14
26
0.09
2.48
|
1194425
447
490
0.18
25
0.10
2.95
|
1194426
320
298
0.21
26
0.10
2.80
|
1194427
147
194
0.18
22
0.08
2.75
|
|
Body weights of CD-1 mice were measured at days 1 and 39, and the average body weight for each group is presented in the table below. Liver, kidney and spleen weights were measured at the end of the study and are presented in the table below. Modified oligonucleotides that caused any changes in organ weights outside the expected range for modified oligonucleotides were excluded from further studies.
TABLE 123
|
|
Body and organ weights (in grams)
|
Compound
Body Weight (g)
Liver
Kidney
Spleen
|
No.
Day 1
Day 39
(g)
(g)
(g)
|
|
Saline
33
38
2.0
0.6
0.1
|
1194402
35
41
2.3
0.6
0.1
|
1194403
32
37
2.2
0.5
0.1
|
1194404
33
39
2.4
0.6
0.2
|
1194405
35
41
2.7
0.6
0.1
|
1194406
34
43
2.7
0.7
0.1
|
1194407
32
38
2.1
0.6
0.1
|
1194408
33
39
2.6
0.7
0.2
|
1194409
32
39
2.7
0.6
0.1
|
1194410
35
41
2.9
0.6
0.2
|
1194411
35
41
2.9
0.6
0.1
|
1194412
32
38
2.7
0.6
0.2
|
1194413
32
39
2.3
0.6
0.1
|
1194415
33
37
2.2
0.6
0.2
|
1194416
34
39
2.3
0.6
0.1
|
1194417
35
41
2.3
0.6
0.2
|
1194419
33
37
2.1
0.5
0.1
|
1194420
33
38
2.2
0.6
0.1
|
1194421
37
42
2.7
0.6
0.1
|
1194423
33
39
2.7
0.6
0.2
|
1194424
33
41
2.8
0.6
0.2
|
1194425
37
41
2.0
0.6
0.1
|
1194426
35
41
2.7
0.6
0.2
|
1194427
34
41
2.9
0.6
0.1
|
|
Study 3
Groups of two 8-to-10-week-old male CD-1 mice were injected subcutaneously once (for a total of 1 treatment) with 50 mg/kg of modified oligonucleotides. One group of two male CD-1 mice was injected with saline. Mice were euthanized 4 days following administration.
To evaluate the effect of modified oligonucleotides on liver and kidney function, plasma levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TBIL), blood urea nitrogen (BUN), creatinine (CRT) and albumin were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400c, Melville, NY). The results are presented in the table below. Assays include two animals in a group, except where the symbol “‡” indicates that only 1 animal was used for a specific assay. Modified oligonucleotides that caused changes in the levels of any of the liver or kidney function markers outside the expected range for modified oligonucleotides were excluded in further studies.
TABLE 124
|
|
Plasma chemistry markers in male CD-1 mice
|
Compound
AST
ALT
TBIL
BUN
CRT
Albumin
|
No.
(IU/L)
(IU/L)
(mg/dL)
(mg/dL)
(mg/dL)
(mg/dL)
|
|
saline
81
63
0.13
18
0.12
2.5
|
1194483
1073
1351
0.78
19
0.13
2.58
|
1194484
5261
6171
4.19
15
0.13
2.46
|
1194486
112
115
0.17
19
0.14
2.85
|
1194487‡
2747
3998
5.21
16
0.2
2.91
|
1194488
68
46
0.14
25
0.14
2.92
|
1194489
43
38
0.16
23
0.13
2.48
|
1194490
61
62
0.13
21
0.13
2.74
|
1194491
59
77
0.16
21
0.16
2.85
|
1194492
47
26
0.13
23
0.14
2.59
|
1194493
114
141
0.15
21
0.15
2.77
|
1194494‡
2255
3189
3.49
25
0.09
2.25
|
1194495
3848
3311
1.48
19
0.15
2.56
|
1194496
839
3491
2.8
21
0.14
2.37
|
1194497
3145
3724
0.77
20
0.17
2.53
|
1194498
229
251
0.17
15
0.12
2.14
|
1194499
519
464
0.93
26
0.18
2.85
|
1194500
58
52
0.16
21
0.12
2.61
|
1194501
60
30
0.13
21
0.14
2.69
|
1194502
45
30
0.15
21
0.14
2.61
|
1194503
57
38
0.14
19
0.14
2.68
|
1194504
47
53
0.15
20
0.13
2.63
|
1194505‡
109
183
0.15
23
0.16
3.06
|
1194506
63
96
0.14
25
0.13
2.86
|
1194507
124
198
0.16
22
0.14
2.41
|
1194508
1291
797
0.3
17
0.14
2.81
|
1194509
6147
5321
3.17
18
0.18
2.99
|
1194510
322
505
0.21
19
0.15
2.83
|
1194512‡
2308
5902
1.98
18
0.09
2.52
|
1194513
87
112
0.21
20
0.14
2.42
|
1194514
60
63
0.13
19
0.14
2.52
|
1194515
90
116
0.14
23
0.13
2.52
|
1194516
54
47
0.15
21
0.16
2.62
|
1194517
102
102
0.14
17
0.16
2.54
|
1194518
78
83
0.15
22
0.13
2.62
|
1194519
43
32
0.19
19
0.12
2.66
|
1194520
84
61
0.12
20
0.15
2.56
|
1194572
148
153
0.14
22
0.11
2.54
|
1194573
71
50
0.16
18
0.14
2.6
|
1194574‡
324
428
0.11
25
0.13
2.77
|
1194575‡
6483
7128
6.54
27
0.14
2.39
|
1194576
2701
1943
6.15
30
0.32
2.44
|
1194578
N/A
1182‡
5.22
85
0.69
2.63
|
1194579
4055
6429
4.34
25
0.18
2.83
|
1194580‡
3302
4048
6.96
24
0.2
2.51
|
1194581
6176
6185
5.21
22
0.21
3.23
|
1194582
1160‡
16‡
3.86
32
0.32
3.31
|
1194583‡
N/A
N/A
5.19
27
0.19
3.44
|
|
Study 4
Groups of four 7-to-8-week-old male CD-1 mice were injected subcutaneously once per week for 6 weeks (for a total of 6 treatments) with 40 mg/kg of modified oligonucleotides. One group of four male CD-1 mice was injected with saline. Mice were euthanized 2 days following final administration.
To evaluate the effect of modified oligonucleotides on liver and kidney function, plasma levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TBIL), blood urea nitrogen (BUN), creatinine (CRT) and albumin were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400c, Melville, NY). The results are presented in the table below. Assays include four animals in a group, except where the symbol “‡” indicates that 3 animals or less was used for a specific assay. Modified oligonucleotides that caused changes in the levels of any of the liver or kidney function markers outside the expected range for modified oligonucleotides were excluded in further studies.
TABLE 125
|
|
Plasma chemistry markers in male CD-1 mice
|
Compound
AST
ALT
TBIL
BUN
CRT
Albumin
|
No.
(IU/L)
(IU/L)
(mg/dL)
(mg/dL)
(mg/dL)
(mg/dL)
|
|
saline
74
57
0.24
20
0.11
3.07
|
1194343
767
1036
0.41
16
0.10
2.87
|
1194345
165
163
0.20
23
0.12
2.75
|
1194346
826
955
0.25
25
0.16
3.06
|
1194347
965
1210
0.22
23
0.12
3.01
|
1194348
1886
3635
0.55
24
0.15
3.15
|
1194349
140
209
0.19
22
0.12
2.75
|
1194350
1342
1861
0.24
23
0.13
2.72
|
1194357
66
46
0.13
25
0.14
2.61
|
1194358
54
59
0.16
24
0.15
2.99
|
1194359
132
202
0.16
22
0.11
2.66
|
1194360
119
263
0.15
19
0.08
2.57
|
1194361
596
740
0.21
27
0.09
2.83
|
1194363‡
441
604
0.19
22
0.11
3.08
|
1194370
502
572
0.16
21
0.09
2.74
|
1194371
58
53
0.18
22
0.09
2.85
|
1194372
510
646
0.27
21
0.09
2.88
|
1194373
677
963
0.24
20
0.08
2.87
|
1194374
1530
2506
0.20
23
0.09
2.99
|
1194375
544
815
0.18
19
0.09
2.81
|
1194376
251
490
0.16
19
0.06
2.54
|
1194377
438
704
0.18
23
0.09
2.75
|
1194429
1282
1188
0.36
21
0.06
2.80
|
1194430
558
820
0.18
20
0.11
2.87
|
|
Body weights of CD-1 mice were measured at days 1 and 35, and the average body weight for each group is presented in the table below. Liver, kidney and spleen weights were measured at the end of the study and are presented in the table below. Modified oligonucleotides that caused any changes in organ weights outside the expected range for modified oligonucleotides were excluded from further studies.
TABLE 126
|
|
Body and organ weights (in grams)
|
Compound
Body Weight (g)
Liver
Kidney
Spleen
|
No.
Day 1
Day 35
(g)
(g)
(g)
|
|
Saline
31
39
2.0
0.6
0.1
|
1194343
31
37
2.3
0.5
0.1
|
1194345
33
40
2.6
0.5
0.1
|
1194346
30
36
2.8
0.5
0.1
|
1194347
34
39
3.1
0.6
0.2
|
1194348
33
37
3.2
0.5
0.1
|
1194349
32
38
2.6
0.5
0.1
|
1194350
32
39
3.3
0.6
0.2
|
1194357
31
36
2.2
0.5
0.2
|
1194358
33
39
2.6
0.5
0.1
|
1194359
31
36
2.3
0.5
0.1
|
1194360
31
36
2.3
0.5
0.2
|
1194361
33
37
2.7
0.5
0.1
|
1194363
31
35
3.4
0.5
0.2
|
1194370
32
36
2.1
0.5
0.2
|
1194371
32
38
2.1
0.5
0.1
|
1194372
31
36
2.3
0.5
0.1
|
1194373
32
36
2.3
0.5
0.1
|
1194374
32
38
3.1
0.6
0.2
|
1194375
31
37
2.9
0.5
0.2
|
1194376
31
37
2.3
0.5
0.1
|
1194377
33
39
2.8
0.5
0.2
|
1194429
31
36
2.7
0.5
0.3
|
1194430
32
36
2.5
0.4
0.1
|
|
Example 11: Tolerability of Modified Oligonucleotides Targeting Human FXII in Sprague-Dawley Rats
Sprague-Dawley rats are a multipurpose model used for safety and efficacy evaluations. The rats were treated with Ionis modified oligonucleotides described in the Examples above and evaluated for changes in the levels of various plasma chemistry markers.
Study 1
Groups of 4 Sprague-Dawley rats each were weekly injected subcutaneously with 80 mg/kg of Ionis oligonucleotide for 6 weeks (total 6 doses). The rats were euthanized; and organs, urine and plasma were harvested for further analysis 3 days after the last dose.
Plasma Chemistry Markers
To evaluate the effect of Ionis oligonucleotides on hepatic function, plasma levels of transaminases were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400c, Melville, NY). Plasma levels of ALT (alanine transaminase) and AST (aspartate transaminase) were measured and the results are presented in the table below expressed in IU/L. Plasma levels of total bilirubin (TBIL), creatinine (CREA), albumin (ALB), and Blood Urea Nitrogen (BUN) were also measured using the same clinical chemistry analyzer and the results are also presented in the table below. Ionis modified oligonucleotides that caused changes in the levels of any markers of liver function outside the expected range for modified oligonucleotides were excluded in further studies. In some cases, where less than 4 samples were available in a group, the compounds are marked with the symbol “‡”.
TABLE 127
|
|
Plasma chemistry markers in Sprague-Dawley rats
|
Compound
AST
ALT
TBIL
BUN
CREA
ALB
|
No.
(IU/L)
(IU/L)
(mg/dL)
(mg/dL)
(mg/dL)
(mg/dL)
|
|
Saline
68
41
0.16
17
0.21
3.13
|
1194380
89
56
0.28
19
0.26
3.03
|
1194383
198
158
0.22
17
0.22
3.18
|
1194384‡
52
24
0.12
17
0.18
2.93
|
1194388
244
80
0.34
17
0.15
2.22
|
1194389
157
103
0.20
27
0.59
3.31
|
1194392
64
29
0.18
18
0.20
3.18
|
1194393
59
32
0.15
16
0.23
3.19
|
1194394
58
28
0.12
17
0.20
3.03
|
1194395
154
83
0.20
17
0.23
2.99
|
1194402
82
64
0.17
19
0.20
3.23
|
1194403
68
37
0.17
16
0.22
3.13
|
|
Blood obtained from rat groups at week 6 were sent to IDEXX BioResearch for measurement of blood cell counts. Counts taken include red blood cell (RBC) count, white blood cell (WBC) count, hemoglobin (HGB), hematocrit (HCT), Mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and individual white blood cell counts, such as that of monocytes (MON), neutrophils (NEU), lymphocytes (LYM), and platelets (PLT). The results are presented in the tables below. Ionis oligonucleotides that caused changes in the blood cell count outside the expected range for modified oligonucleotides were excluded in further studies.
TABLE 128
|
|
Blood Cell Count in Sprague-Dawley Rats
|
Compound
WBC
RBC
HGB
HCT
MCV
MCH
MCHC
NEU
LYM
MON
PLT
|
No.
(/nL)
(/pL)
(g/dL)
(%)
(fL)
(pg)
(%)
(%)
(%)
(%)
(/nL)
|
|
Saline
11
8
15
44
52
18
35
17
78
4
786
|
1194380
5
9
15
44
52
18
34
16
77
5
981
|
1194383
10
9
17
48
51
18
35
12
80
5
916
|
1194384
8
9
15
44
51
18
35
15
79
4
958
|
1194388
6
5
10
34
68
21
31
19
74
6
229
|
1194389
10
9
16
44
52
18
36
16
79
4
1080
|
1194392
7
8
15
44
53
18
35
13
83
3
792
|
1194393
7
9
16
46
53
18
35
19
74
5
714
|
1194394
5
8
14
41
54
18
34
8
88
2
513
|
1194395
10
9
15
42
49
17
35
13
80
5
792
|
1194402
8
9
16
45
50
17
35
9
86
3
800
|
1194403
6
9
15
44
51
18
35
17
78
4
939
|
|
To evaluate the effect of Ionis oligonucleotides on kidney function, urinary levels of micro total protein (MTP) and creatinine were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400c, Melville, NY). The ratios of MTP to creatinine (MTP/C ratio) are presented in the table below. Ionis oligonucleotides that caused changes in the levels of the ratio outside the expected range for modified oligonucleotides were excluded in further studies.
TABLE 129
|
|
MTP to creatinine ratio in Sprague-Dawley rats
|
Compound No.
MTP/C Ratio
|
|
saline
1.7
|
1194380
1.9
|
1194383
2.5
|
1194384
2.5
|
1194388
2.4
|
1194389
1.9
|
1194392
1.6
|
1194393
1.7
|
1194394
1.9
|
1194395
1.9
|
1194402
2.0
|
1194403
1.8
|
|
Body weights of rats were measured at days 1 and 38, and the average body weight for each group is presented in the table below. Liver, spleen and kidney weights were measured at the end of the study, and are presented in the table below. Ionis oligonucleotides that caused any changes in organ weights outside the expected range for modified oligonucleotides were excluded from further studies.
TABLE 130
|
|
Body and organ weights (g)
|
Compound
Body Weight (g)
Liver
Kidney
Spleen
|
No.
Day 1
Day 38
Weight (g)
Weight (g)
Weight (g)
|
|
Saline
266
424
18
3.4
1.0
|
1194380
255
378
17
3.1
1.1
|
1194383
260
370
16
3.3
1.2
|
1194384
256
396
17
3.5
1.4
|
1194388
256
346
27
3.4
4.8
|
1194389
257
398
18
3.3
1.4
|
1194392
256
387
19
3.3
1.5
|
1194393
245
363
16
2.8
1.0
|
1194394
265
389
18
3.5
1.9
|
1194395
264
377
14
3.2
1.3
|
1194402
249
335
13
2.7
1.1
|
1194403
260
387
16
3.4
1.1
|
|
Study 2
Groups of 4 Sprague-Dawley rats each were weekly injected subcutaneously with 80 mg/kg of Ionis oligonucleotide for 6 weeks (total 6 doses). The rats were euthanized; and organs, urine and plasma were harvested for further analysis 3 days after the last dose.
Plasma Chemistry Markers
To evaluate the effect of Ionis oligonucleotides on hepatic function, plasma levels of transaminases were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400c, Melville, NY). Plasma levels of ALT (alanine transaminase) and AST (aspartate transaminase) were measured and the results are presented in the table below expressed in IU/L. Plasma levels of total bilirubin (TBIL), creatinine (CREA), albumin (ALB), and Blood Urea Nitrogen (BUN) were also measured using the same clinical chemistry analyzer and the results are also presented in the table below. Ionis modified oligonucleotides that caused changes in the levels of any markers of liver function outside the expected range for modified oligonucleotides were excluded in further studies. In some cases, where less than 4 samples were available in a group, the compounds are marked with the symbol “‡”.
TABLE 131
|
|
Plasma chemistry markers in Sprague-Dawley rats
|
Compound
AST
ALT
TBIL
BUN
CREA
ALB
|
No.
(IU/L)
(IU/L)
(mg/dL)
(mg/dL)
(mg/dL)
(mg/dL)
|
|
Saline
81
49
0.16
15
0.22
3.11
|
1194404
99
47.5
0.22
18.375
0.28
3.21
|
1194405
853
783
0.28
19
0.30
3.32
|
1194406
96
50
0.16
16
0.21
3.40
|
1194407
71
34
0.15
17
0.24
3.20
|
1194408
223
179
0.23
16
0.24
3.49‡
|
1194409
169
133
0.24
17
0.22
3.30
|
1194413‡
209
167
0.28
17
0.21
3.27
|
1194415
61
41
0.14
17
0.21
3.44
|
1194416‡
91
57
0.16
17
0.22
3.47
|
1194419
105
48
0.16
15
0.23
3.32
|
1194420
84
52
0.13
16
0.22
3.36
|
1194427
113
71
0.18
14
0.22
3.65‡
|
|
Blood obtained from rat groups at week 6 were sent to IDEXX BioResearch for measurement of blood cell counts. Counts taken include red blood cell (RBC) count, white blood cell (WBC) count, hemoglobin (HGB), hematocrit (HCT), Mean corpuscular volume (MCV) mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and individual white blood cell counts, such as that of monocytes (MON), neutrophils (NEU), lymphocytes (LYM), and platelets (PLT). The results are presented in the tables below. Ionis oligonucleotides that caused changes in the blood cell count outside the expected range for modified oligonucleotides were excluded in further studies.
TABLE 132
|
|
Blood Cell Count in Sprague-Dawley Rats
|
Compound
WBC
RBC
HGB
HCT
MCV
MCH
MCH
NEU
LYM
MON
PLT
|
No.
(/nL)
(/pL)
(g/dL)
(%)
(fL)
(pg)
C (%)
(%)
(%)
(%)
(/nL)
|
|
Saline
12
8
15
43
55
19
35
17
78
5
882
|
1194404
13
8
15
43
53
19
35
10
86
4
868
|
1194405
10
9
16
44
52
18
35
10
77
12
909
|
1194406
11
8
15
44
52
18
35
11
82
6
867
|
1194407
11
8
15
44
54
19
35
12
83
4
817
|
1194408
9
8
16
45
54
19
35
14
78
7
918
|
1194409
10
8
15
43
55
19
35
8
86
6
735
|
1194413‡
8
8
15
44
55
19
35
12
79
8
798
|
1194415
8
8
16
45
54
19
35
10
82
7
855
|
1194416
9
8
16
45
53
19
35
10
83
6
1093
|
1194419
5
8
15
42
54
19
35
11
81
8
813
|
1194420
8
9
16
45
53
19
35
13
80
5
835
|
1194427
12
8
16
47
56
19
35
13
80
6
1003
|
|
To evaluate the effect of Ionis oligonucleotides on kidney function, urinary levels of micro total protein (MTP) and creatinine were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400c, Melville, NY). The ratios of MTP to creatinine (MTP/C ratio) are presented in the table below. Ionis oligonucleotides that caused changes in the levels of the ratio outside the expected range for modified oligonucleotides were excluded in further studies.
TABLE 133
|
|
MTP to creatinine ratio in Sprague-Dawley rats
|
Compound No.
MTP/C Ratio
|
|
saline
0.8
|
1194404
1.1
|
1194405
1.6
|
1194406
1.5
|
1194407
1.8
|
1194408
1.9
|
1194409
1.5
|
1194413
1.4
|
1194415
2.0
|
1194416
1.4
|
1194419
2.0
|
1194420
2.3
|
1194427
2.6
|
|
Body weights of rats were measured at days 1 and 35, and the average body weight for each group is presented in the table below. Liver, spleen and kidney weights were measured at the end of the study, and are presented in the table below. Ionis oligonucleotides that caused any changes in organ weights outside the expected range for modified oligonucleotides were excluded from further studies.
TABLE 134
|
|
Body and organ weights (g)
|
Compound
Body Weight (g)
Liver
Kidney
Spleen
|
No.
Day 1
Day 35
Weight (g)
Weight (g)
Weight (g)
|
|
Saline
219
433
17
3.8
4.4
|
1194404
213
391
15
3.3
5.5
|
1194405
210
357
16
3.3
4.4
|
1194406
218
388
16
3.6
4.7
|
1194407
227
410
17
3.9
5.0
|
1194408
222
390
15
3.6
4.8
|
1194409
216
376
16
3.7
5.3
|
1194413*
207
399
17
4.0
4.2
|
1194415
220
382
15
3.5
3.9
|
1194416
218
369
13
3.3
4.7
|
1194419
218
388
16
3.2
3.6
|
1194420
219
392
16
3.6
4.6
|
1194427
204
375
15
3.2
4.0
|
|
Study 3
Groups of 4 Sprague-Dawley rats each were weekly injected subcutaneously with 40 mg/kg of Ionis oligonucleotide for 6 weeks (total 6 doses). A group of 4 rats were treated subcutaneously with 80 mg/kg of Compound No. 1194391. The rats were euthanized; and organs, urine and plasma were harvested for further analysis 2 days after the last dose.
Plasma Chemistry Markers
To evaluate the effect of Ionis oligonucleotides on hepatic function, plasma levels of transaminases were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400c, Melville, NY). Plasma levels of ALT (alanine transaminase) and AST (aspartate transaminase) were measured and the results are presented in the table below expressed in IU/L. Plasma levels of total bilirubin (TBIL), creatinine (CREA), albumin (ALB), and Blood Urea Nitrogen (BUN) were also measured using the same clinical chemistry analyzer and the results are also presented in the table below. Ionis modified oligonucleotides that caused changes in the levels of any markers of liver function outside the expected range for modified oligonucleotides were excluded in further studies.
TABLE 135
|
|
Plasma chemistry markers in Sprague-Dawley rats
|
Compound
AST
ALT
TBIL
BUN
CREA
ALB
|
No.
(IU/L)
(IU/L)
(mg/dL)
(mg/dL)
(mg/dL)
(mg/dL)
|
|
Saline
65
52
0.16
15
0.23
3.31
|
1194345
122
52
0.15
19
0.25
2.81
|
1194349
59
38
0.16
16
0.27
3.54
|
1194357
78
50
0.16
16
0.28
3.67
|
1194358
82
51
0.18
16
0.29
3.88
|
1194359
70
41
0.15
15
0.28
3.65
|
1194360
269
92
0.35
20
0.27
2.66
|
1194371
125
85
0.20
17
0.25
2.74
|
1194377
1054
319
1.04
16
0.35
3.18
|
1194430
1195
379
1.04
16
0.26
3.39
|
1194391
83
65
0.17
15
0.21
3.39
|
|
Blood obtained from rat groups at week 6 were sent to IDEXX BioResearch for measurement of blood cell counts. Counts taken include red blood cell (RBC) count, white blood cell (WBC) count, hemoglobin (HGB), hematocrit (HCT), Mean corpuscular volume (MCV) mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and individual white blood cell counts, such as that of monocytes (MON), neutrophils (NEU), lymphocytes (LYM), and platelets (PLT). The results are presented in the tables below. Ionis oligonucleotides that caused changes in the blood cell count outside the expected range for modified oligonucleotides were excluded in further studies.
TABLE 136
|
|
Blood Cell Count in Sprague-Dawley Rats
|
Compound
WBC
RBC
HGB
HCT
MCV
MCH
MCH
NEU
LYM
MON
PLT
|
No.
(/nL)
(/pL)
(g/dL)
(%)
(fL)
(pg)
C (%)
(%)
(%)
(%)
(/nL)
|
|
Saline
9
8
16
47
58
19
34
16
81
2
749
|
1194345
12
7
14
42
61
20
33
9
87
3
687
|
1194349
10
8
15
45
55
19
34
21
75
3
921
|
1194357
9
8
16
47
57
19
34
10
86
3
814
|
1194358
11
9
16
47
55
19
34
13
82
4
1020
|
1194359
10
8
15
45
55
19
34
13
82
3
887
|
1194360
16
8
13
38
50
17
34
10
84
6
733
|
1194371
9
8
14
43
56
18
33
19
74
5
910
|
1194377
9
8
15
43
56
19
34
17
76
6
917
|
1194430
13
8
14
42
56
19
34
20
73
7
1241
|
1194391
8
8
16
46
58
20
34
22
74
3
803
|
|
To evaluate the effect of Ionis oligonucleotides on kidney function, urinary levels of micro total protein (MTP) and creatinine were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400c, Melville, NY). The ratios of MTP to creatinine (MTP/C ratio) are presented in the table below. Ionis oligonucleotides that caused changes in the levels of the ratio outside the expected range for modified oligonucleotides were excluded in further studies.
TABLE 137
|
|
MTP to creatinine ratio in Sprague-Dawley rats
|
Compound No.
MTP/C Ratio
|
|
saline
1.1
|
1194345
1.8
|
1194349
2.9
|
1194357
2.1
|
1194358
1.6
|
1194359
1.8
|
1194360
1.7
|
1194371
1.2
|
1194377
2.4
|
1194430
2.2
|
1194391
1.4
|
|
Body weights of rats were measured at days 1 and 38, and the average body weight for each group is presented in the table below. Liver, spleen and kidney weights were measured at the end of the study, and are presented in the table below. Ionis oligonucleotides that caused any changes in organ weights outside the expected range for modified oligonucleotides were excluded from further studies.
TABLE 138
|
|
Body and organ weights (g)
|
Compound
Body Weight (g)
Liver
Kidney
Spleen
|
No.
Day 1
Day 38
Weight (g)
Weight (g)
Weight (g)
|
|
Saline
237
450
17
3.5
0.8
|
1194345
249
446
20
3.8
2.3
|
1194349
232
425
19
3.2
1.2
|
1194357
235
409
17
3.6
1.2
|
1194358
240
396
16
3.3
1.2
|
1194359
249
414
15
3.4
1.1
|
1194360
239
385
15
3.5
1.8
|
1194371
235
422
19
3.5
1.4
|
1194377
243
376
10
3.0
1.4
|
1194391
233
414
16
3.3
0.8
|
1194430
243
406
13
3.7
1.5
|
|
Study 4
Groups of 4 Sprague-Dawley rats each were weekly injected subcutaneously with 40 mg/kg of Ionis oligonucleotide for 6 weeks (total 6 doses). The rats were euthanized; and organs, urine and plasma were harvested for further analysis 3 days after the last dose.
Plasma Chemistry Markers
To evaluate the effect of Ionis oligonucleotides on hepatic function, plasma levels of transaminases were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400c, Melville, NY). Plasma levels of ALT (alanine transaminase) and AST (aspartate transaminase) were measured and the results are presented in the table below expressed in IU/L. Plasma levels of total bilirubin (TBIL), creatinine (CREA), albumin (ALB), and Blood Urea Nitrogen (BUN) were also measured using the same clinical chemistry analyzer and the results are also presented in the table below. Ionis modified oligonucleotides that caused changes in the levels of any markers of liver function outside the expected range for modified oligonucleotides were excluded in further studies.
TABLE 139
|
|
Plasma chemistry markers in Sprague-Dawley rats
|
Compound
AST
ALT
TBIL
BUN
CREA
ALB
|
No.
(IU/L)
(IU/L)
(mg/dL)
(mg/dL)
(mg/dL)
(mg/dL)
|
|
Saline
61
43
0.1
16
0.3
3.4
|
1270699
86
63
0.2
17
0.3
3.7
|
1270700
195
157
0.3
17
0.4
3.6
|
1270701
83
47
0.2
15
0.3
3.6
|
1270703
113
61
0.2
25
0.4
3.5
|
1270705
254
246
0.2
17
0.3
3.5
|
1270707
213
196
0.2
18
0.3
3.6
|
1270710
130
61
0.1
17
0.3
3.5
|
1270712
119
80
0.2
17
0.3
3.6
|
1270714
112
70
0.2
22
0.3
3.3
|
1270715
337
198
0.3
19
0.3
3.5
|
1270716
195
92
0.2
15
0.3
3.6
|
|
Blood obtained from rat groups at week 6 were sent to IDEXX BioResearch for measurement of blood cell counts. Counts taken include red blood cell (RBC) count, white blood cell (WBC) count, hemoglobin (HGB), hematocrit (HCT), Mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and individual white blood cell counts, such as that of monocytes (MON), neutrophils (NEU), lymphocytes (LYM), and platelets (PLT). The results are presented in the tables below. Ionis oligonucleotides that caused changes in the blood cell count outside the expected range for modified oligonucleotides were excluded in further studies.
TABLE 140
|
|
Blood Cell Count in Sprague-Dawley Rats
|
Compound
WBC
RBC
HGB
HCT
MCV
MCH
MCH
NEU
LYM
MON
PLT
|
No.
(/nL)
(/pL)
(g/dL)
(%)
(fL)
(pg)
C (%)
(%)
(%)
(%)
(/nL)
|
|
Saline
6
9
16
47
54
18
33
15
78
6
804
|
1270699
7
9
15
45
52
18
34
11
82
6
717
|
1270700
11
9
15
44
51
18
35
10
83
6
813
|
1270701
8
9
15
44
51
18
34
13
80
6
814
|
1270703
9
8
14
43
54
18
33
19
73
6
757
|
1270705
9
9
15
45
51
18
34
15
76
8
799
|
1270707
10
9
16
45
52
18
35
10
81
8
939
|
1270710
13
8
13
39
49
17
34
12
81
6
778
|
1270712
7
8
14
42
51
17
34
21
69
8
780
|
1270714
9
9
16
44
50
18
36
19
72
8
754
|
1270715
13
8
14
42
51
17
34
11
83
5
865
|
1270716
8
8
14
42
51
17
34
11
80
8
738
|
|
To evaluate the effect of Ionis oligonucleotides on kidney function, urinary levels of micro total protein (MTP) and creatinine were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400c, Melville, NY). The ratios of MTP to creatinine (MTP/C ratio) are presented in the table below. Ionis oligonucleotides that caused changes in the levels of the ratio outside the expected range for modified oligonucleotides were excluded in further studies.
TABLE 141
|
|
MTP to creatinine ratio in Sprague-Dawley rats
|
Compound No.
MTP/C Ratio
|
|
saline
1.0
|
1270699
2.5
|
1270700
1.0
|
1270701
1.6
|
1270703
1.3
|
1270705
1.2
|
1270707
1.8
|
1270710
2.6
|
1270712
1.2
|
1270714
0.9
|
1270715
3.1
|
1270716
2.5
|
|
Body weights of rats were measured at days 1 and 35, and the average body weight for each group is presented in the table below. Liver, spleen and kidney weights were measured at the end of the study, and are presented in the table below. Ionis oligonucleotides that caused any changes in organ weights outside the expected range for modified oligonucleotides were excluded from further studies.
TABLE 142
|
|
Body and organ weights (g)
|
Compound
Body Weight (g)
Liver
Kidney
Spleen
|
No.
Day 1
Day 35
Weight (g)
Weight (g)
Weight (g)
|
|
Saline
469
529
18
4.0
0.8
|
1270699
484
511
22
4.3
1.6
|
1270700
496
507
19
4.0
1.4
|
1270701
469
513
19
4.4
1.2
|
1270703
491
509
19
4.6
1.4
|
1270705
499
527
21
4.7
1.6
|
1270707
486
500
21
4.5
1.7
|
1270710
499
540
22
4.6
1.5
|
1270712
500
552
17
4.5
1.4
|
1270714
504
548
17
4.8
1.2
|
1270715
474
494
19
4.0
1.7
|
1270716
477
501
17
4.2
1.5
|
|
Example 12: Activity of Modified Oligonucleotides Complementary to Human FXII in Transgenic Mice, Single Dose
A transgenic mouse model was developed in-house using the Fosmid NCBI Clone DB ID: ABC12-49040200E6. The clone was digested at AgeI and NdeI restriction sites to produce a region containing the entire ˜23 kb portion of human F12 gene, including regions 12 kb upstream and 4 kb downstream of the F12 gene. The gene fragment was introduced into fertilized eggs from C57Bl/6 mice by pronuclear injection to produce FXII founder lines. Line 16459 was used in the experiments described herein. Human FXII RNA expression is found in the liver and circulating protein is found in plasma.
Study 1
Treatment
The transgenic mice were divided into groups of 2 female mice each for modified oligonucleotide treatment. Groups received subcutaneous injections of modified oligonucleotide at a dose of 2.5 mg/kg twice a week for two weeks (4 treatments). One group of two mice received subcutaneous injections of PBS twice a week for two weeks (4 treatments). The PBS-injected group served as the control group to which oligonucleotide-treated groups were compared.
RNA Analysis
48 hours after the last treatment, mice were sacrificed and RNA was extracted from liver for real-time RTPCR analysis of FXII RNA expression. Human FXII primer probe sets RTS2992 (described herein above) and RTS40528 (described herein above) were used to measure human FXII RNA levels. FXII RNA levels were normalized to total RNA content, as measured by RIBOGREEN®. Reduction of FXII RNA is presented in the tables below as percent FXII RNA relative to the amount in PBS control animals (% control).
TABLE 143
|
|
Reduction of human FXII RNA in transgenic mice
|
Compound
FXII RNA (% control) in liver
|
No.
RTS2992
RTS40528
|
|
PBS
100
100
|
1194343
2
9
|
1194347
15
20
|
|
Protein Analysis
Human FXII protein levels in plasma were determined using an Abcam FXII ELISA kit (ab192144). Reduction of FXII protein is presented in the tables below as percent FXII protein relative to the amount in PBS control animals (% control).
TABLE 144
|
|
Reduction of human FXII protein in transgenic mice
|
Compound No.
FXII protein (% control) in plasma
|
|
PBS
100
|
1194343
0.3
|
1194347
17
|
|
Study 2
Treatment
The transgenic mice were divided into groups of 4 mice (2 male and 2 female) each for modified oligonucleotide treatment. Groups received subcutaneous injections of modified oligonucleotide at a dose of 1.5 mg/kg twice a week for two weeks (4 treatments). One group of four mice (2 male and 2 female) received subcutaneous injections of PBS twice a week for two weeks (4 treatments). The PBS-injected group served as the control group to which oligonucleotide-treated groups were compared.
RNA Analysis
48 hours after the last treatment, mice were sacrificed and RNA was extracted from liver for real-time RTPCR analysis of FXII RNA expression. Human FXII primer probe sets RTS2992 (described herein above) and RTS40528 (described herein above) were used to measure human FXII RNA levels. FXII RNA levels were normalized to total RNA content, as measured by RIBOGREEN®. Reduction of FXII RNA is presented in the tables below as percent FXII RNA relative to the amount in PBS treated animals (% control).
TABLE 145
|
|
Reduction of human FXII RNA in transgenic mice
|
FXII RNA (%
|
Compound
control) in liver
|
No.
RTS2992
RTS40528
|
|
PBS
100
100
|
1194380
32
36
|
1194383
52
58
|
1194384
32
35
|
1194389
54
76
|
1194391
45
57
|
1194392
48
68
|
1194393
37
40
|
1194394
31
41
|
1194395
21
35
|
1194402
22
27
|
1194403
12
10
|
1194404
17
17
|
1194406
31
40
|
1194407
56
76
|
1194408
21
23
|
1194409
22
20
|
1194413
82
80
|
1194415
83
111
|
1194416
81
96
|
1194419
45
54
|
1194420
61
53
|
1194427
90
71
|
1194345
14
12
|
1194349
10
9
|
1194357
3
3
|
1194358
3
2
|
1194359
6
4
|
1194360
5
4
|
1194371
5
6
|
1194377
12
12
|
1194430
21
22
|
|
Protein Analysis
Human FXII protein levels in plasma were determined using an Abcam FXII ELISA kit (ab192144). Reduction of FXII protein is presented in the tables below as percent FXII protein relative to the amount in PBS treated animals (% control).
TABLE 146
|
|
Reduction of human FXII protein in transgenic mice
|
Compound No.
FXII protein (% control) in plasma
|
|
PBS
100
|
1194380
23
|
1194383
41
|
1194384
24
|
1194389
56
|
1194391
36
|
1194392
38
|
1194393
27
|
1194394
20
|
1194395
12
|
1194402
14
|
1194403
4
|
1194404
9
|
1194406
23
|
1194407
42
|
1194408
16
|
1194409
15
|
1194413
61
|
1194415
80
|
1194416
72
|
1194419
42
|
1194420
61
|
1194427
87
|
1194345
11
|
1194349
3
|
1194357
1
|
1194358
0
|
1194359
1
|
1194360
2
|
1194371
1
|
1194377
9
|
1194430
20
|
|
Example 13: Activity of Modified Oligonucleotides Complementary to Human FXII in Transgenic Mice, Multiple Dose
A transgenic mouse model was developed in-house using the Fosmid NCBI Clone DB ID: ABC12-49040200E6. The clone was digested at AgeI and NdeI restriction sites to produce a region containing the entire ˜23 kb portion of human F12 gene, including regions 12 kb upstream and 4 kb downstream of the F12 gene. The gene fragment was introduced into fertilized eggs from C57Bl/6 mice by pronuclear injection to produce FXII founder lines. Line 16459 was used in the experiments described herein. Human FXII RNA expression is found in the liver and circulating protein is found in plasma.
Study 1
Treatment
The transgenic mice were divided into groups of 2 female mice each for modified oligonucleotide treatment. Groups received subcutaneous injections of modified oligonucleotide at doses indicated in the table below twice a week for two weeks (4 treatments). One group of two mice received subcutaneous injections of PBS twice a week for two weeks (4 treatments). The PBS-injected group served as the control group to which oligonucleotide-treated groups were compared.
RNA Analysis
48 hours after the last treatment, mice were sacrificed and RNA was extracted from liver for real-time RTPCR analysis of FXII RNA expression. Human FXII primer probe set RTS2992 (described herein above) was used to measure human FXII RNA levels. FXII RNA levels were normalized to total RNA content, as measured by RIBOGREEN®. Reduction of FXII RNA is presented in the tables below as percent FXII RNA relative to the amount in PBS treated animals (% control).
TABLE 147
|
|
Reduction of human FXII RNA in transgenic mice
|
RTS2992
|
Compound
Concentration
FXII RNA (%
ED50
|
No.
(mpk)
control) in liver
(mpk)
|
|
1194347
0.32
75
1.292
|
0.8
59
|
2
50
|
5
14
|
|
Protein Analysis
Human FXII protein levels in plasma were determined using an Abcam FXII ELISA kit (ab192144). Reduction of FXII protein is presented in the tables below as percent FXII protein relative to the amount in PBS treated animals (% control).
TABLE 148
|
|
Reduction of human FXII protein in transgenic mice
|
Compound
Concentration
FXII protein (% control)
ED50
|
No.
(mpk)
in plasma
(mpk)
|
|
1194347
0.32
78
1.073
|
0.8
64
|
2
31
|
5
11
|
|
Study 2
Treatment
The transgenic mice were divided into groups of four mice (2 male and 2 female) each for modified oligonucleotide treatment. Groups received subcutaneous injections of modified oligonucleotide at doses indicated in the table below twice a week for two weeks (4 treatments). One group of four mice (2 male and 2 female) received subcutaneous injections of PBS twice a week for two weeks (4 treatments). The PBS-injected group served as the control group to which oligonucleotide-treated groups were compared.
RNA Analysis
48 hours after the last treatment, mice were sacrificed and RNA was extracted from liver for real-time RTPCR analysis of FXII RNA expression. Human FXII primer probe set RTS2992 (described herein above) was used to measure human FXII RNA levels. FXII RNA levels were normalized to total RNA content, as measured by RIBOGREEN®. Reduction of FXII RNA is presented in the tables below as percent FXII RNA relative to the amount in PBS treated animals (% control).
TABLE 149
|
|
Reduction of human FXII RNA in transgenic mice
|
FXII RNA
|
(%
|
control) in
|
Compound
Concentration
liver
ED50
|
No.
(mpk)
(RTS2992)
(mpk)
|
|
1194357
0.08
60
0.07514
|
0.1
33
|
0.2
27
|
0.3
14
|
0.6
8
|
0.9
4
|
1194349
0.1
60
0.4603
|
0.3
61
|
0.9
25
|
1194358
0.1
47
0.08653
|
0.3
18
|
0.9
9
|
1194359
0.1
80
0.1997
|
0.3
36
|
0.9
9
|
1194371
0.1
114
0.2737
|
0.3
35
|
0.9
10
|
1194377
0.1
91
0.5223
|
0.3
70
|
0.9
28
|
1270699
0.1
79
0.1731
|
0.3
31
|
0.9
6
|
1270700
0.1
57
0.1211
|
0.3
21
|
0.9
4
|
1270701
0.1
38
0.1871
|
0.3
41
|
0.9
6
|
1270702
0.1
59
0.1816
|
0.3
22
|
0.9
6
|
1270703
0.08
76
0.2228
|
0.2
71
|
0.6
11
|
1270705
0.08
62
0.113
|
0.2
33
|
0.6
12
|
1270707
0.08
82
0.2085
|
0.2
57
|
0.6
14
|
1270710
0.08
92
0.265
|
0.2
61
|
0.6
19
|
1270712
0.08
92
0.3297
|
0.2
63
|
0.6
14
|
1270714
0.08
57
0.3936
|
0.2
47
|
0.6
25
|
1270715
0.08
69
0.3242
|
0.2
57
|
0.6
20
|
1270716
0.08
58
0.3093
|
0.2
52
|
0.6
20
|
|
Protein Analysis
Human FXII protein levels in plasma were determined using an Abcam FXII ELISA kit (ab192144). Reduction of FXII protein is presented in the tables below as percent FXII protein relative to the amount in PBS treated animals (% control).
TABLE 150
|
|
Reduction of human FXII protein in transgenic mice
|
FXII
|
protein
|
(%
|
control)
|
Compound
Concentration
in
ED50
|
No.
(mpk)
plasma
(mpk)
|
|
1194357
0.08
67
0.065
|
0.1
43
|
0.2
25
|
0.3
15
|
0.6
3
|
0.9
6
|
1194349
0.1
84
0.2799
|
0.3
48
|
0.9
13
|
1194358
0.1
52
0.09148
|
0.3
20
|
0.9
5
|
1194359
0.1
53
0.0959
|
0.3
22
|
0.9
3
|
1194371
0.1
65
0.1356
|
0.3
29
|
0.9
3
|
1194377
0.1
80
0.2882
|
0.3
50
|
0.9
16
|
1270699
0.1
62
0.1153
|
0.3
22
|
0.9
4
|
1270700
0.1
53
0.08752
|
0.3
17
|
0.9
1
|
1270701
0.1
42
0.08131
|
0.3
27
|
0.9
5
|
1270702
0.1
65
0.1331
|
0.3
27
|
0.9
4
|
1270703
0.08
79
0.1572
|
0.2
41
|
0.6
7
|
1270705
0.08
66
0.1174
|
0.2
36
|
0.6
7
|
1270707
0.08
77
0.1668
|
0.2
46
|
0.6
8
|
1270710
0.08
86
0.2288
|
0.2
56
|
0.6
13
|
1270712
0.08
86
0.2467
|
0.2
58
|
0.6
16
|
1270714
0.08
93
0.3053
|
0.2
62
|
0.6
24
|
1270715
0.08
89
0.3739
|
0.2
62
|
0.6
37
|
1270716
0.08
89
0.3447
|
0.2
71
|
0.6
25
|
|
Study 3
Treatment
The transgenic mice were divided into groups of four mice (2 male and 2 female) each for modified oligonucleotide treatment. Groups received subcutaneous injections of modified oligonucleotide at doses indicated in the table below twice a week for two weeks (4 treatments). One group of four mice (2 male and 2 female) received subcutaneous injections of PBS twice a week for two weeks (4 treatments). The PBS-injected group served as the control group to which oligonucleotide-treated groups were compared.
RNA Analysis
72 hours after the last treatment, mice were sacrificed and RNA was extracted from liver for real-time RTPCR analysis of FXII RNA expression. Human FXII primer probe sets RTS2992 (described herein above) was used to measure human FXII RNA levels. FXII RNA levels were normalized to total RNA content, as measured by RIBOGREEN®. Reduction of FXII RNA is presented in the tables below as percent FXII RNA relative to the amount in PBS treated animals (% control).
TABLE 151
|
|
Reduction of human FXII RNA in transgenic mice
|
RTS2992
|
Compound
Concentration
FXII RNA
ED50
|
No.
(mpk)
(% control) in liver
(mpk)
|
|
1194371
0.075
94
0.2457
|
0.15
57
|
0.3
47
|
0.6
22
|
1.2
10
|
|
Example 14: Effect of Modified Oligonucleotides on Human FXII mRNA In Vitro, Multiple Doses
Modified oligonucleotides selected from the examples above were tested at various doses in Huh7 cells. Cultured Huh7 cells at a density of 20,000 cells per well were transfected by electroporation with various concentrations of modified oligonucleotide as specified in the table below. After a treatment period of approximately 24 hours, total RNA was isolated from the cells, and FXII mRNA levels were measured by quantitative real-time RTPCR. Human FXII primer probe set RTS40528 (described herein above) was used to measure RNA levels. FXII levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Reduction of FXII RNA is presented in the tables below as percent FXII RNA relative to the amount in untreated control cells (% UTC). The half maximal inhibitory concentration (IC50) of each modified oligonucleotide was calculated in GraphPad Prism using the log (inhibitor) vs. normalized response—variable slope method.
Four comparator compounds from WO 2017/120397 were added to the study. These include Compound Nos. 1213034, 1358035, 1213030 and 1213029 designated as A-145676, A-145669, A-145668, and A-145667 respectively in WO 2017/120397.
TABLE 152
|
|
Dose-dependent reduction of human FXII RNA by modified
|
oligonucleotides
|
Compound
FXII RNA (% UTC)
IC50
|
No.
10 μM
2 μM
0.4 μM
0.08 μM
0.016 μM
(μM)
|
|
1129485
7
15
44
81
98
0.34
|
1129486
8
18
51
96
97
0.48
|
1207006
8
10
28
52
84
0.1
|
1207160
17
18
30
58
65
0.09
|
1207161
12
20
27
68
83
0.17
|
1207277
6
9
29
59
84
0.13
|
1207279
12
29
45
80
95
0.43
|
1207393
7
14
26
63
83
0.14
|
1213260
12
18
26
62
87
0.15
|
1213144
8
25
37
71
95
0.28
|
Comparator Compounds
|
1213030
43
84
132
132
94
8.15
|
1213034
69
116
139
127
92
>10
|
1213029
63
123
146
130
98
>10
|
1358035
43
93
127
121
104
8.59
|
|
Patent Grant
12018261
