Patent Application
20240360447
The contents of the electronic sequence listing (PAT059574_Sequence_Listing_ST26.xml; Size: 1,512,836 bytes; and Date of Creation: May 9, 2024) are hereby incorporated by reference in their entirety.
The present disclosure relates to compounds and methods for the treatment of Charcot-Marie-Tooth disease. More specifically, the present disclosure relates to inhibitors of PMP22 and their use in the treatment of Charcot-Marie-Tooth disease.
Charcot-Marie-Tooth (CMT) disease is an inherited peripheral neuropathy characterized by slowly progressive muscle atrophy. CMT is one of the most common inherited neurological disorders, affecting approximately 150,000 people across the United States and Europe. There are several subtypes of CMT disease, each having a distinct genetic cause. The most common form of CMT, accounting for as many as 60% of cases, is CMT type 1A (CMT1A), which results from an excess of peripheral myelin protein 22 (PMP22) protein due to the duplication of one PMP22 alelle.
The PMP22 protein is a major component of myelin that comprises between two and five percent of the myelin that insulates peripheral nerves. While the exact role of PMP22 is not known, there is evidence that overexpression of PMP22 alters the growth and differentiation of Schwann cells, the cells responsible for producing the myelin sheath around neurons. The myelin sheath is a protective layer of lipids and proteins that serves as insulation around nerve axons and facilitates the ability to rapidly conduct nerve signals. In addition to causing deficiencies in the ability to generate new myelin, the presence of excess PMP22 protein in the myelin sheath has been reported to directly destabilize the myelin sheath, leading to increased rates of demyelination. Defects in the myelin sheath reduce the speed that nerve signals can be propagated along nerves, known as the motor nerve conduction velocity, or MNCV. This in turn leads to progressive muscle atrophy in the peripheral limbs resulting in muscle weakness, structural abnormalities in the feet, and abnormal spinal curvature.
Overexpression of PMP22 in mice results in symptoms characteristic of CMT1A disease, including muscle weakness, gait abnormalities, myelination defects, and reduced nerve conduction velocities. Under the control of a conditionally regulated promoter, PMP22 overexpression caused demyelination of neurons, which was reversed upon subsequent suppression of PMP22 expression. Within one week, new myelin sheath formation was evident and within 12 weeks, myelinated neurons were similar to those present in transgenic mice in which PMP22 expression was not suppressed.
Mice harboring three to four copies of the human PMP22 gene develop pathologies similar to those observed in subjects with CMT1A and as such, these mice are used as an experimental model of CMT1A. In this model, treatment with an antisense oligonucleotide complementary to human PMP22 lowered PMP22 mRNA levels and led to restoration of myelination, improvement of MNCV and reversal of other neuropathy endpoints. However, the high doses required in the mouse model translate to dosages that are unlikely to be tolerated in human subjects, thus antisense oligonucleotides targeted to PMP22 have not advanced to development as a treatment for CMT1A.
While a small number of potential therapies are being evaluated in clinical trials, an effective treatment for any CMT disease, including CMT1A, has yet to be identified. Current care consists of physical therapy, occupational therapy and orthopedic devices to help patients cope with disabling symptoms, and pain-relieving drugs for patients with severe pain. Accordingly, there remains an unmet medical need for therapeutic agents for the treatment of CMT1A.
Provided herein are, inter alia, nucleic acid compounds targeted to the peripheral myelin protein 22 (PMP22) mRNA.
In embodiments, provided is a compound comprising an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein each of the antisense strand and sense strands is 15 to 25 nucleotides in length, the nucleotide sequence of the antisense strand is at least 90% complementary to the nucleotide sequence of the PMP22 mRNA (SEQ ID NO: 1170), and the nucleotide sequence of the sense strand has no more than two mismatches to the nucleotide sequence of the antisense strand.
In embodiments, each of the antisense strand and sense strands is 15 to 25 nucleotides in length, the nucleotide sequence of the antisense strand comprises at least 15 contiguous nucleotides of any one of SEQ ID NOs 491, 492, 493, 494, 495, 497, 498, 503, 504, 506, 510, 511, 514, 515, 516, 518, 524, 526, 529, 531, 532, 533, 534, 535, 536, 538, 539, 540, 541, 542, 543, 545, 546, 547, 548, 550, 553, 554, 556, 558, 559, 560, 561, 563, 567, 569, 575, 576, 579, 580, 581, 582, 583, 585, 590, 591, 595, 597, 600, 605, 609, 610, 618, 622, 623, 628, 630, 631, 633, 635, 637, 639, 641, 642, 643, 644, 645, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1122, 1124, 1125, 1126, 1127, 1128, 1129, 1130, 1131, 1132, 1133, 1134, 1135, 1136, 1137, 1138, 1139, 1140, 1141, 1142, 1143, 1144, 1145, 1146, 1147, 1148, 1149, 1150, 1151, 1152, 1153, 1154, 1155, 1156, 1157, 1158, 1159, 1160, 1161, 1162, 1163, 1164, 1165, 1166, 1167, 1168, 1169, 1118, 1121, 1123, 1126, and 1144, and the nucleotide sequence of the sense strand has no more than two mismatches to the nucleotide sequence of the antisense strand.
In embodiments, the antisense strand and the sense strand are not covalently linked.
In embodiments, at least one nucleotide of the antisense strand is a modified nucleotide. In embodiments, at least one nucleotide of the sense strand is a modified nucleotide. In embodiments, the 5′-terminal nucleotide of the antisense strand comprises a 5′-VP modification.
In embodiments, the antisense strand is 21 to 23 nucleotides in length. In embodiments, the sense strand is 21 to 23 nucleotides in length.
In embodiments, the hybridization of the antisense strand to the sense strand forms at least one blunt end. In embodiments, at least one strand comprises a 3′ nucleotide overhang of one to five nucleotides.
In embodiments, the compound comprises a ligand covalently linked to the antisense strand or the sense strand.
In embodiments, the compound has the structure:
A is the sense strand or the antisense strand. t is an integer from 1 to 5.
L3 and L4 are independently a bond, —N(R23)—, —O—, —S—, —C(O)—, —N(R23)C(O)—, —C(O)N(R24)—, —N(R23)C(O)N(R24)—, —C(O)O—, —OC(O)—, —N(R23)C(O)O—, —OC(O)N(R24)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R25)—O—, —O—P(S)(R25)—O—, —O—P(O)(NR23R24)—N—, —O—P(S)(NR23R24)—N—, —O—P(O)(NR23R24)—O—, —O—P(S)(NR23R24)—O—, —P(O)(NR23R24)—N—, —P(S)(NR23R24)—N—, —P(O)(NR23R24)—O—, —P(S)(NR23R24)—O—, —S—S—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene or substituted or unsubstituted heteroarylene. Each R23, R24 and R25 is independently hydrogen or unsubstituted C1-C10 alkyl.
L5 is -L5A-L5B-L5C-L5D-L5E-. L6 is -L6A-L6B-L6C-L6D-L6E-. L5A, L5B, L5C, L5D, L5E, L6A, L6B L6C, L6D, and L6E are independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene or substituted or unsubstituted heteroarylene; and each R23, R24 and R25 is independently hydrogen or unsubstituted C1-C10 alkyl.
R1 and R2 are independently unsubstituted C1-C25 alkyl, wherein at least one of R1 and R2 is unsubstituted C9-C19 alkyl. R3 is hydrogen, —NH2, —OH, —SH, —C(O)H, —C(O)NH2, —NHC(O)H, —NHC(O)OH, —NHC(O)NH2, —C(O)OH, —OC(O)H, —N3, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
In embodiments, provided herein is a pharmaceutical composition comprising the compound as described herein.
In embodiments, provided herein are methods for inhibiting the expression of peripheral myelin protein 22 (PMP22) mRNA in a cell, comprising contacting the cell with a compound of provided herein, thereby inhibiting the expression of PMP22 mRNA in the cell
In embodiments, provided herein are methods for inhibiting the expression of peripheral myelin protein 22 (PMP22) mRNA in a subject, comprising administering to the subject an effective amount of a compound or pharmaceutical composition provided herein, thereby inhibiting the expression of peripheral myelin protein 22 (PMP22) mRNA.
In embodiments, provided herein are methods for increasing myelination and/or slowing the loss of myelination in a subject, comprising administering to the subject an effective amount of a compound or pharmaceutical composition provided herein.
In embodiments, provided herein are methods for treating Charcot-Marie-Tooth disease (CMT) in a subject, comprising administering to the subject an effective amount of a compound or pharmaceutical composition provided herein. In embodiments, the Charcot-Marie-Tooth disease (CMT) is Charcot-Marie-Tooth disease Type 1A (CMT1A).
Unless defined otherwise, all technical terms, scientific terms, abbreviations, chemical structures, and chemical formulae used herein have the same meaning as is commonly understood by one of ordinary skill in the art. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts. All patents, applications, published applications, and other publications referenced herein are incorporated by reference in their entirety unless stated otherwise. Unless otherwise indicated, conventional methods of mass spectroscopy, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA techniques, and pharmacology are employed. Furthermore, use of the term “including” as well as other forms, such as “include”, “includes,” and “included,” is not limiting. As used in this specification, whether in a transitional phrase or in the body of the claim, the terms “comprise(s)” and “comprising” are to be interpreted as having an open-ended meaning. That is, the terms are to be interpreted synonymously with the phrases “having at least” or “including at least.” When used in the context of a process, the term “comprising” means that the process includes at least the recited steps, but may include additional steps. When used in the context of a compound, composition, or device, the term “comprising” means that the compound, composition, or device includes at least the recited features or components, but may also include additional features or components.
Where substituent groups are specified by their conventional chemical formulae, written from left to right, they equally encompass the chemically identical substituents that would result from writing the structure from right to left, e.g., —CH2O— is equivalent to —OCH2—.
“Charcot-Marie-Tooth disease” or “CMT” means an inherited peripheral neuropathy affecting both motor and sensory nerves. CMT is characterized by muscle weakness and atrophy in the legs and arms, foot deformities and loss of sensation and/or numbness. CMT disease includes the CMT1A subtype, among others.
“Charcot-Marie-Tooth disease Type 1A” or CMT1A means the subtype of CMT that results from a duplication of one PMP22 allele, resulting in three copies of the PMP22 gene in subjects.
“Nerve conduction velocity” means the speed with which an electrical impulse moves through a nerve. In embodiments, nerve conduction velocity is motor nerve conduction velocity. In embodiments, nerve conduction velocity is sensory nerve conduction velocity. In embodiments, nerve conduction velocity may be determined by an electroneuroagraphy, i.e. a nerve conduction study.
“Compound muscle action potential” is a is a quantitative measure of the amplitude of the electrical impulses that are transmitted to muscle, correlating with the number of muscle fibers that can be activated. In embodiments, compound muscle action potential is determined by electromyography (EMG).
“Improve” means to lessen the severity of a symptom and/or clinical indicator of a disease.
“Slow the progression of” means to reduce the rate at which a symptom and/or clinical indicator of a disease becomes more severe.
“Therapeutically effective amount” means an amount sufficient for a compound to provide a therapeutic benefit to a subject.
“Subject” used herein means a human or non-human animal selected for treatment or therapy. In embodiments, a subject is a human.
“Administration” means providing a pharmaceutical agent or composition to a subject, and includes administration performed by a medical professional and self-administration. In embodiments, administration is intravenous administration. In embodiments, administration is subcutaneous administration.
“Treating” or “treatment” means the administration of one or more pharmaceutical agents to a subject to achieve a desired clinical result, including but not limited to the alleviation, improvement, or slowing of the progression of at least one clinical indicator and/or symptom of a disease in a subject.
“Delay the onset of” means to delay the development of a condition or disease in a subject who is at risk for developing the disease or condition. In embodiments, a subject at risk for developing a disease or condition is identified using clinical assessments similar to those used to diagnose the disease or condition. For example, a subject at risk for developing CMT1A may be identified by genetic testing for amplication of the PMP22 gene. In embodiments, a subject at risk for developing the disease or condition receives treatment similar to the treatment received by a subject who already has the disease or condition.
“Effective amount” means an amount sufficient for a compound that, when administered to a subject, is sufficient to effect treatment of a disease in the subject. An effective amount may vary depending on one or more of the potency of the compound, its mode of administration, the severity of the disease in the subject, concomitant pharmaceutical agents the subject is receiving, and characteristics of the subject such as the subject's medical history, age, and weight.
“Pharmaceutical salt” means a salt form of a compound that retains the biological effectiveness and properties of a compound and does not have undesired effects when administered to a subject.
“Compound” means a molecule comprising linked monomeric nucleotides. A compound may have one or more modified nucleotides. In embodiments, a compound comprises a double-stranded nucleic acid. In embodiments, a compound comprises a single-stranded nucleic acid. A compound may be provided as a pharmaceutical salt. A compound may be provided as a pharmaceutical composition.
“Oligonucleotide” means a polymer of linked monomeric nucleotides. One or more nucleotides of an oligonucleotide may be a modified nucleotide.
“Double-stranded nucleic acid” means a first nucleotide sequence hybridized to a second nucleotide sequence to form a duplex structure. Double-stranded nucleic acids include structures formed from annealing a first oligonucleotide to a second, complementary oligonucleotide, as in an siRNA. Such double-stranded nucleic acids may have a short nucleotide overhang at one or both ends of the duplex structure. Double-stranded nucleic acids also include structures formed from a single oligonucleotide with sufficient length and self-complementarity to form a duplex structure, as in an shRNA. Such double-stranded nucleic acids include stem-loop structures. A double-stranded nucleic acid may include one or more modifications relative to a naturally occurring terminus, sugar, nucleobase, and/or phosphate group.
“Double-stranded region” means the portion of a double-stranded nucleic acid where nucleotides of the first nucleotide sequence are hybridized to nucleotides of the second nucleotide sequence. A double-stranded region can be a defined portion within a double-stranded nucleic acid that is shorter than (e.g. encompassed by) the full double-stranded nucleic acid. Alternatively, a double-stranded region can be the same length as the full double-stranded nucleic acid. A double-stranded region may contain one or more mismatches between the first and second nucleotide sequences, and retain the ability hybridize with each other. Double-stranded regions do not include nucleotide overhangs.
“Antisense strand” means an oligonucleotide that is complementary to a target RNA (e.g. a mRNA) and is incorporated into the RNA-induced silencing complex (RISC) to direct gene silencing in a sequence-specific manner through the RNA interference pathway. The antisense strand may also be referred to as the “guide strand.”
“Sense strand” means an oligonucleotide that is complementary to the antisense strand of a double-stranded nucleic acid. The sense strand is typically degraded following incorporation of the antisense strand into RISC. The sense strand may also be referred to as the “passenger strand.”
“Nucleotide overhang” means an extension of one or more unpaired nucleotides from the double-stranded region of a double-stranded nucleic acid. For example, when the 3′ terminus of an antisense strand extends beyond the 5′ terminus of a sense strand, the 3′ terminus of the antisense strand has a nucleotide overhang. A nucleotide overhang can be one, two, three, four or five nucleotides. One or more nucleotides of a nucleotide overhang may be a modified nucleotide. A nucleotide overhang may be on the antisense strand, the sense strand, or both the antisense and sense strands.
“Blunt end” means a given terminus of a double-stranded nucleic acid with no unpaired nucleotides extending from the double-stranded region, i.e. there is no nucleotide overhang. A double-stranded nucleic acid may have a blunt end at one or both termini.
“siRNA” means a double-stranded nucleic acid formed from separate antisense and sense strands, which directs gene silencing in a sequence-specific manner by facilitating mRNA degradation before translation through the RNA interference pathway. The antisense and sense strands of an siRNA are not covalently linked.
“shRNA” means a double-stranded nucleic acid containing a loop structure that is processed in a cell to an siRNA which directs gene silencing in a sequence-specific manner, by facilitating mRNA degradation before translation through the RNA interference pathway.
“Single-stranded nucleic acid” means an antisense strand that is not hybridized to a complementary strand. A single-stranded nucleic acid is incorporated into RISC to direct gene silencing in a sequence-specific manner by facilitating mRNA degradation before translation through the RNA interference pathway.
“Hybridize” means the annealing of one nucleotide sequence to another nucleotide sequence based at least in part on nucleotide sequence complementarity. In embodiments, an antisense strand is hybridized to a sense strand. In embodiments, an antisense strand hybridizes to a target mRNA sequence.
“Complementary” means nucleobases having the capacity to pair non-covalently via hydrogen bonding.
“Fully complementary” or “100% complementary” means each nucleobase of a first nucleotide sequence is complementary to each nucleobase of a second nucleotide sequence. In embodiments, an antisense strand is fully complementary to its target mRNA. In embodiments, a sense strand and an antisense strand of double-stranded nucleic acid are fully complementary over their entire lengths. In embodiments, a sense strand and an antisense strand of double-stranded nucleic acid are fully complementary over the entire length of the double-stranded region of the siRNA, and one or both termini of either strand comprises single-stranded nucleotides.
“Percent complementary” means the percentage of nucleobases of an oligonucleotide that are complementary to an equal-length portion of a target nucleic acid. Percent complementarity is calculated by dividing the number of nucleobases of the oligonucleotide that are complementary to nucleobases at corresponding positions in the target nucleic acid by the total number of nucleobases in the oligonucleotide.
“Identical” in the context of nucleotide sequences, means having the same nucleotide sequence, independent of sugar, linkage, and/or nucleobase modifications and independent of the methylation state of any pyrimidines present.
“Percent identity” means the number of nucleobases in a first nucleotide sequence that are identical to nucleobases at corresponding positions in a second nucleotide sequence, divided by the total number of nucleobases in the first nucleotide sequence.
“Mismatch” means a nucleobase of a first nucleotide sequence that is not capable of Watson-Crick pairing with a nucleobase at a corresponding position of a second nucleotide sequence.
“Nucleoside” means a monomer of a nucleobase and a pentofuranosyl sugar (e.g., either ribose or deoxyribose). Nucleosides may comprise bases such as A, C, G, T, or U, or modifications thereof. Nucleosides may be modified at the base and/or and the sugar. In embodiments, a nucleoside is a deoxyribonucleoside. In embodiments, the nucleoside is a ribonucleoside.
“Nucleotide” means a nucleoside covalently linked to a phosphate group at the 5′ carbon of the pentafuranosyl sugar. Nucleotides may be modified at one or more of the nucleobase, sugar moiety, internucleotide linkage and/or phosphate group.
“Nucleobase” means a heterocyclic base moiety capable of non-covalently pairing. Nucleobases include pyrimidines and purines. Unless stated otherwise, conventional nucleobase abbreviations are used herein. Nucleobases abbreviations include, without limitation, A (adenine), C (cytosine), G (guanine), T (thymine), U (uracil).
Unless stated otherwise, numbering of nucleotide atoms is according to standard numbering convention, with the carbons of the pentafuranosyl sugar numbered 1′ through 5′, and the nucleobase atoms numbered 1 through 9 for purines and 1 through 6 for pyrimidines.
“Modified nucleoside” means a nucleoside having one or more modifications relative to a naturally occurring nucleoside. Such alterations may be present in a nucleobase and/or sugar moiety of the nucleoside. A modified nucleoside may have a modified sugar moiety and an unmodified nucleobase. A modified nucleoside may have a modified sugar moiety and a modified nucleobase.
“Modified nucleotide” means a nucleotide having one or more alterations relative to a naturally occurring nucleotide. An alteration may be present in an internucleoside linkage, a nucleobase, and/or a sugar moiety of the nucleotide. A modified nucleotide may have a modified sugar moiety and an unmodified phosphate group. A modified nucleotide may have an unmodified sugar moiety and a modified phosphate group. A modified nucleotide may have a modified sugar moiety and an unmodified nucleobase. A modified nucleotide may have a modified sugar moiety and a modified phosphate group.
“Modified nucleobase” means a nucleobase having one or more alterations relative to a naturally occurring nucleobase.
“Modified phosphate group” means any change from a naturally occurring phosphate group of a nucleotide.
“Modified internucleotide linkage” means any change from a naturally occurring phosphodiester linkage between two nucleotides.
“Phosphorothioate internucleotide linkage” means a substituted phosphodiester internucleotide linkage where one of the non-bridging atoms is a sulfur atom.
“Modified sugar moiety” means a sugar of a nucleotide having any change and/or substitution from a naturally occurring sugar moiety.
“beta-D-deoxyribonucleoside” means a naturally occurring nucleoside monomer of DNA.
“beta-D-ribonucleoside” means a naturally occurring nucleoside monomer of RNA.
“2′-O-methyl sugar” or “2′-OMe sugar” means a sugar having an O—CH3 substitution at the 2′ position of the pentofuranosyl sugar.
“2′-O-methoxyethyl sugar” or “2′-MOE sugar” means a sugar having an OCH2CH2OCH3 substitution at the 2′ position of the pentofuranosyl sugar.
“2-fluoro sugar” or “2′-F sugar” means a sugar having a fluoro substitution at the 2′ position of the pentofuranosyl sugar.
“Bicyclic sugar” means a modified sugar moiety comprising a linkage connecting the 2′-carbon and 4′-carbon of the pentafuranosyl sugar, resulting in a bicyclic structure. Nonlimiting exemplary bicyclic sugar moieties include LNA, ENA, cEt, S-cEt, and R-cEt.
“Locked nucleic acid (LNA) sugar” means a substituted sugar moiety comprising a —CH2—O— linkage between the 4′ and 2′ furanose ring atoms.
“ENA sugar” means a substituted sugar moiety comprising a —(CH2)2—O— linkage between the 4′ and 2′ furanose ring atoms.
“2′-O-methyl nucleotide” means a nucleotide having an O-methyl substitution at the 2′ position of the pentofuranosyl sugar. A 2′-O-methyl nucleotide may have a further modification in addition to the modified sugar moiety, for example a modified nucleobase and/or phosphate group.
“2′-fluoro nucleotide” means a nucleotide having a fluoro substitution at the 2′ position of the pentofuranosyl sugar. A 2′-O-fluoro nucleotide may have a further modification in addition to the modified sugar moiety, for example a modified nucleobase and/or phosphate group.
“Bicyclic nucleotide” means a nucleotide having a linkage connecting the 2′-carbon and 4′-carbon of the pentafuranosyl sugar. A bicyclic nucleotide may have a further modification in addition to the modified sugar moiety, for example a modified nucleobase and/or phosphate group.
“5′-(E)-vinylphosphonate” or “5′-VP”, refers to a chemical moiety having the structure:
or salts thereof, where the wavy line represent the point of attachment to the 5′ carbon of the pentafuranosyl sugar of a nucleotide.
“5-methylcytosine” means a cytosine nucleobase having a 5-methyl substitution on the cytosine ring.
“Non-methylated cytosine” means a cytosine nucleobase that does not have a methyl substitution at the 5 position of the cytosine ring.
“5-methyluracil” means a uracil nucleobase having a 5-methyl substitution on the uracil ring. A 5-methyluracil nucleobase may also be referred to as a thymine.
“Non-methylated uracil” means a uracil nucleobase that does not have a methyl group substitution at the 5 position of the uracil ring.
The term “alkyl,” by itself or as part of another substituent, means, unless otherwise stated, a straight (i.e., unbranched) or branched carbon chain (or carbon), or combination thereof, which may be fully saturated, mono- or polyunsaturated and can include mono-, di- and multivalent radicals. The alkyl may include a designated number of carbons (e.g., C1-C10 means one to ten carbons). Alkyl is an uncyclized chain. Examples of saturated hydrocarbon radicals include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, methyl, homologs and isomers of, for example, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. An unsaturated alkyl group is one having one or more double bonds or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and the higher homologs and isomers. An alkoxy is an alkyl attached to the remainder of the molecule via an oxygen linker (—O—). An alkyl moiety may be an alkenyl moiety. An alkyl moiety may be an alkynyl moiety. An alkyl moiety may be fully saturated. An alkenyl may include more than one double bond and/or one or more triple bonds in addition to the one or more double bonds. An alkynyl may include more than one triple bond and/or one or more double bonds in addition to the one or more triple bonds.
The term “cycloalkyl” means a monocyclic, bicyclic, or a multicyclic cycloalkyl ring system. In embodiments, monocyclic ring systems are cyclic hydrocarbon groups containing from 3 to 8 carbon atoms, where such groups can be saturated or unsaturated, but not aromatic. In embodiments, cycloalkyl groups are fully saturated. Examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl. Bicyclic cycloalkyl ring systems are bridged monocyclic rings or fused bicyclic rings. In embodiments, bridged monocyclic rings contain a monocyclic cycloalkyl ring where two non adjacent carbon atoms of the monocyclic ring are linked by an alkylene bridge of between one and three additional carbon atoms (i.e., a bridging group of the form (CH2)w, where w is 1, 2, or 3). Representative examples of bicyclic ring systems include, but are not limited to, bicyclo[3.1.1]heptane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane, bicyclo[3.3.1]nonane, and bicyclo[4.2.1]nonane. In embodiments, fused bicyclic cycloalkyl ring systems contain a monocyclic cycloalkyl ring fused to either a phenyl, a monocyclic cycloalkyl, a monocyclic cycloalkenyl, a monocyclic heterocyclyl, or a monocyclic heteroaryl. In embodiments, the bridged or fused bicyclic cycloalkyl is attached to the parent molecular moiety through any carbon atom contained within the monocyclic cycloalkyl ring. In embodiments, cycloalkyl groups are optionally substituted with one or two groups which are independently oxo or thia. In embodiments, the fused bicyclic cycloalkyl is a 5 or 6 membered monocyclic cycloalkyl ring fused to either a phenyl ring, a 5 or 6 membered monocyclic cycloalkyl, a 5 or 6 membered monocyclic cycloalkenyl, a 5 or 6 membered monocyclic heterocyclyl, or a 5 or 6 membered monocyclic heteroaryl, wherein the fused bicyclic cycloalkyl is optionally substituted by one or two groups which are independently oxo or thia. In embodiments, multicyclic cycloalkyl ring systems are a monocyclic cycloalkyl ring (base ring) fused to either (i) one ring system selected from the group consisting of a bicyclic aryl, a bicyclic heteroaryl, a bicyclic cycloalkyl, a bicyclic cycloalkenyl, and a bicyclic heterocyclyl; or (ii) two other ring systems independently selected from the group consisting of a phenyl, a bicyclic aryl, a monocyclic or bicyclic heteroaryl, a monocyclic or bicyclic cycloalkyl, a monocyclic or bicyclic cycloalkenyl, and a monocyclic or bicyclic heterocyclyl. In embodiments, the multicyclic cycloalkyl is attached to the parent molecular moiety through any carbon atom contained within the base ring. In embodiments, multicyclic cycloalkyl ring systems are a monocyclic cycloalkyl ring (base ring) fused to either (i) one ring system selected from the group consisting of a bicyclic aryl, a bicyclic heteroaryl, a bicyclic cycloalkyl, a bicyclic cycloalkenyl, and a bicyclic heterocyclyl; or (ii) two other ring systems independently selected from the group consisting of a phenyl, a monocyclic heteroaryl, a monocyclic cycloalkyl, a monocyclic cycloalkenyl, and a monocyclic heterocyclyl. Examples of multicyclic cycloalkyl groups include, but are not limited to tetradecahydrophenanthrenyl, perhydrophenothiazin-1-yl, and perhydrophenoxazin-1-yl.
In embodiments, a cycloalkyl is a cycloalkenyl. The term “cycloalkenyl” is used in accordance with its plain ordinary meaning. In embodiments, a cycloalkenyl is a monocyclic, bicyclic, or a multicyclic cycloalkenyl ring system. In embodiments, monocyclic cycloalkenyl ring systems are cyclic hydrocarbon groups containing from 3 to 8 carbon atoms, where such groups are unsaturated (i.e., containing at least one annular carbon carbon double bond), but not aromatic. Examples of monocyclic cycloalkenyl ring systems include cyclopentenyl and cyclohexenyl. In embodiments, bicyclic cycloalkenyl rings are bridged monocyclic rings or a fused bicyclic rings. In embodiments, bridged monocyclic rings contain a monocyclic cycloalkenyl ring where two non adjacent carbon atoms of the monocyclic ring are linked by an alkylene bridge of between one and three additional carbon atoms (i.e., a bridging group of the form (CH2)w, where w is 1, 2, or 3). Representative examples of bicyclic cycloalkenyls include, but are not limited to, norbornenyl and bicyclo[2.2.2]oct 2 enyl. In embodiments, fused bicyclic cycloalkenyl ring systems contain a monocyclic cycloalkenyl ring fused to either a phenyl, a monocyclic cycloalkyl, a monocyclic cycloalkenyl, a monocyclic heterocyclyl, or a monocyclic heteroaryl. In embodiments, the bridged or fused bicyclic cycloalkenyl is attached to the parent molecular moiety through any carbon atom contained within the monocyclic cycloalkenyl ring. In embodiments, cycloalkenyl groups are optionally substituted with one or two groups which are independently oxo or thia. In embodiments, multicyclic cycloalkenyl rings contain a monocyclic cycloalkenyl ring (base ring) fused to either (i) one ring system selected from the group consisting of a bicyclic aryl, a bicyclic heteroaryl, a bicyclic cycloalkyl, a bicyclic cycloalkenyl, and a bicyclic heterocyclyl; or (ii) two ring systems independently selected from the group consisting of a phenyl, a bicyclic aryl, a monocyclic or bicyclic heteroaryl, a monocyclic or bicyclic cycloalkyl, a monocyclic or bicyclic cycloalkenyl, and a monocyclic or bicyclic heterocyclyl. In embodiments, the multicyclic cycloalkenyl is attached to the parent molecular moiety through any carbon atom contained within the base ring. In embodiments, multicyclic cycloalkenyl rings contain a monocyclic cycloalkenyl ring (base ring) fused to either (i) one ring system selected from the group consisting of a bicyclic aryl, a bicyclic heteroaryl, a bicyclic cycloalkyl, a bicyclic cycloalkenyl, and a bicyclic heterocyclyl; or (ii) two ring systems independently selected from the group consisting of a phenyl, a monocyclic heteroaryl, a monocyclic cycloalkyl, a monocyclic cycloalkenyl, and a monocyclic heterocyclyl.
In embodiments, a heterocycloalkyl is a heterocyclyl. The term “heterocyclyl” as used herein, means a monocyclic, bicyclic, or multicyclic heterocycle. The heterocyclyl monocyclic heterocycle is a 3, 4, 5, 6 or 7 membered ring containing at least one heteroatom independently selected from the group consisting of O, N, and S where the ring is saturated or unsaturated, but not aromatic. The 3 or 4 membered ring contains 1 heteroatom selected from the group consisting of O, N and S. The 5 membered ring can contain zero or one double bond and one, two or three heteroatoms selected from the group consisting of O, N and S. The 6 or 7 membered ring contains zero, one or two double bonds and one, two or three heteroatoms selected from the group consisting of O, N and S. The heterocyclyl monocyclic heterocycle is connected to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the heterocyclyl monocyclic heterocycle. Representative examples of heterocyclyl monocyclic heterocycles include, but are not limited to, azetidinyl, azepanyl, aziridinyl, diazepanyl, 1,3 dioxanyl, 1,3 dioxolanyl, 1,3 dithiolanyl, 1,3 dithianyl, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolinyl, isoxazolidinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl, oxazolinyl, oxazolidinyl, piperazinyl, piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, thiadiazolinyl, thiadiazolidinyl, thiazolinyl, thiazolidinyl, thiomorpholinyl, 1,1 dioxidothiomorpholinyl (thiomorpholine sulfone), thiopyranyl, and trithianyl. The heterocyclyl bicyclic heterocycle is a monocyclic heterocycle fused to either a phenyl, a monocyclic cycloalkyl, a monocyclic cycloalkenyl, a monocyclic heterocycle, or a monocyclic heteroaryl. The heterocyclyl bicyclic heterocycle is connected to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the monocyclic heterocycle portion of the bicyclic ring system. Representative examples of bicyclic heterocyclyls include, but are not limited to, 2,3 dihydrobenzofuran 2 yl, 2,3 dihydrobenzofuran 3 yl, indolin 1 yl, indolin 2 yl, indolin 3 yl, 2,3 dihydrobenzothien 2 yl, decahydroquinolinyl, decahydroisoquinolinyl, octahydro 1H indolyl, and octahydrobenzofuranyl. In embodiments, heterocyclyl groups are optionally substituted with one or two groups which are independently oxo or thia. In certain embodiments, the bicyclic heterocyclyl is a 5 or 6 membered monocyclic heterocyclyl ring fused to a phenyl ring, a 5 or 6 membered monocyclic cycloalkyl, a 5 or 6 membered monocyclic cycloalkenyl, a 5 or 6 membered monocyclic heterocyclyl, or a 5 or 6 membered monocyclic heteroaryl, wherein the bicyclic heterocyclyl is optionally substituted by one or two groups which are independently oxo or thia. Multicyclic heterocyclyl ring systems are a monocyclic heterocyclyl ring (base ring) fused to either (i) one ring system selected from the group consisting of a bicyclic aryl, a bicyclic heteroaryl, a bicyclic cycloalkyl, a bicyclic cycloalkenyl, and a bicyclic heterocyclyl; or (ii) two other ring systems independently selected from the group consisting of a phenyl, a bicyclic aryl, a monocyclic or bicyclic heteroaryl, a monocyclic or bicyclic cycloalkyl, a monocyclic or bicyclic cycloalkenyl, and a monocyclic or bicyclic heterocyclyl. The multicyclic heterocyclyl is attached to the parent molecular moiety through any carbon atom or nitrogen atom contained within the base ring. In embodiments, multicyclic heterocyclyl ring systems are a monocyclic heterocyclyl ring (base ring) fused to either (i) one ring system selected from the group consisting of a bicyclic aryl, a bicyclic heteroaryl, a bicyclic cycloalkyl, a bicyclic cycloalkenyl, and a bicyclic heterocyclyl; or (ii) two other ring systems independently selected from the group consisting of a phenyl, a monocyclic heteroaryl, a monocyclic cycloalkyl, a monocyclic cycloalkenyl, and a monocyclic heterocyclyl. Examples of multicyclic heterocyclyl groups include, but are not limited to 10H-phenothiazin-10-yl, 9,10-dihydroacridin-9-yl, 9,10-dihydroacridin-10-yl, 10H-phenoxazin-10-yl, 10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl, 1,2,3,4-tetrahydropyrido[4,3-g]isoquinolin-2-yl, 12H-benzo[b]phenoxazin-12-yl, and dodecahydro-1H-carbazol-9-yl.
The term “alkylene,” by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from an alkyl, as exemplified, but not limited by, —CH2CH2CH2CH2—. Typically, an alkyl (or alkylene) group will have from 1 to 24 carbon atoms, with those groups having 10 or fewer carbon atoms being preferred herein. A “lower alkyl” or “lower alkylene” is a shorter chain alkyl or alkylene group, generally having eight or fewer carbon atoms. The term “alkenylene,” by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from an alkene.
The term “heteroalkyl,” by itself or in combination with another term, means, unless otherwise stated, a stable straight or branched chain, or combinations thereof, including at least one carbon atom and at least one heteroatom (e.g., O, N, S, Si, or P), and wherein the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. The heteroatom(s) (e.g., O, N, S, Si, or P) may be placed at any interior position of the heteroalkyl group or at the position at which the alkyl group is attached to the remainder of the molecule. Heteroalkyl is an uncyclized chain. Examples include, but are not limited to: —CH2—CH2—O—CH3, —CH2—CH2—NH—CH3, —CH2—CH2—N(CH3)—CH3, —CH2—S—CH2—CH3, —CH2—CH2, —S(O)—CH3, —CH2—CH2—S(O)2—CH3, —CH═CH—O—CH3, —Si(CH3)3, —CH2—CH═N—OCH3, —CH═CH—N(CH3)—CH3, —O—CH3, —O—CH2—CH3, and —CN. Up to two or three heteroatoms may be consecutive, such as, for example, —CH2—NH—OCH3 and —CH2—O—Si(CH3)3. A heteroalkyl moiety may include one heteroatom (e.g., O, N, S, Si, or P). A heteroalkyl moiety may include two optionally different heteroatoms (e.g., O, N, S, Si, or P). A heteroalkyl moiety may include three optionally different heteroatoms (e.g., O, N, S, Si, or P). A heteroalkyl moiety may include four optionally different heteroatoms (e.g., O, N, S, Si, or P). A heteroalkyl moiety may include five optionally different heteroatoms (e.g., O, N, S, Si, or P). A heteroalkyl moiety may include up to 8 optionally different heteroatoms (e.g., O, N, S, Si, or P). The term “heteroalkenyl,” by itself or in combination with another term, means, unless otherwise stated, a heteroalkyl including at least one double bond. A heteroalkenyl may optionally include more than one double bond and/or one or more triple bonds in additional to the one or more double bonds. The term “heteroalkynyl,” by itself or in combination with another term, means, unless otherwise stated, a heteroalkyl including at least one triple bond. A heteroalkynyl may optionally include more than one triple bond and/or one or more double bonds in additional to the one or more triple bonds.
Similarly, the term “heteroalkylene,” by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from heteroalkyl, as exemplified, but not limited by, —CH2—CH2—S—CH2—CH2— and —CH2—S—CH2—CH2—NH—CH2—. For heteroalkylene groups, heteroatoms can also occupy either or both of the chain termini (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, and the like). Still further, for alkylene and heteroalkylene linking groups, no orientation of the linking group is implied by the direction in which the formula of the linking group is written. For example, the formula —C(O)2R′— represents both —C(O)2R′— and —R′C(O)2—. As described above, heteroalkyl groups, as used herein, include those groups that are attached to the remainder of the molecule through a heteroatom, such as —C(O)R′, —C(O)NR′, —NR′R″, —OR′, —SR′, and/or —SO2R′. Where “heteroalkyl” is recited, followed by recitations of specific heteroalkyl groups, such as —NR′R″ or the like, it will be understood that the terms heteroalkyl and —NR′R″ are not redundant or mutually exclusive. Rather, the specific heteroalkyl groups are recited to add clarity. Thus, the term “heteroalkyl” should not be interpreted herein as excluding specific heteroalkyl groups, such as —NR′R″ or the like.
The terms “cycloalkyl” and “heterocycloalkyl,” by themselves or in combination with other terms, mean, unless otherwise stated, cyclic versions of “alkyl” and “heteroalkyl,” respectively. Cycloalkyl and heterocycloalkyl are not aromatic. Additionally, for heterocycloalkyl, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples of heterocycloalkyl include, but are not limited to, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl, and the like. A “cycloalkylene” and a “heterocycloalkylene,” alone or as part of another substituent, means a divalent radical derived from a cycloalkyl and heterocycloalkyl, respectively.
The terms “halo” or “halogen,” by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as “haloalkyl” are meant to include monohaloalkyl and polyhaloalkyl. For example, the term “halo(C1-C4)alkyl” includes, but is not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.
The term “acyl” means, unless otherwise stated, —C(O)R where R is a substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
The term “aryl” means, unless otherwise stated, a polyunsaturated, aromatic, hydrocarbon substituent, which can be a single ring or multiple rings (preferably from 1 to 3 rings) that are fused together (i.e., a fused ring aryl) or linked covalently. A fused ring aryl refers to multiple rings fused together wherein at least one of the fused rings is an aryl ring.
The term “heteroaryl” refers to aryl groups (or rings) that contain at least one heteroatom such as N, O, or S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized. Thus, the term “heteroaryl” includes fused ring heteroaryl groups (i.e., multiple rings fused together wherein at least one of the fused rings is a heteroaromatic ring). A 5,6-fused ring heteroarylene refers to two rings fused together, wherein one ring has 5 members and the other ring has 6 members, and wherein at least one ring is a heteroaryl ring. Likewise, a 6,6-fused ring heteroarylene refers to two rings fused together, wherein one ring has 6 members and the other ring has 6 members, and wherein at least one ring is a heteroaryl ring. And a 6,5-fused ring heteroarylene refers to two rings fused together, wherein one ring has 6 members and the other ring has 5 members, and wherein at least one ring is a heteroaryl ring. A heteroaryl group can be attached to the remainder of the molecule through a carbon or heteroatom. Non-limiting examples of aryl and heteroaryl groups include phenyl, naphthyl, pyrrolyl, pyrazolyl, pyridazinyl, triazinyl, pyrimidinyl, imidazolyl, pyrazinyl, purinyl, oxazolyl, isoxazolyl, thiazolyl, furyl, thienyl, pyridyl, pyrimidyl, benzothiazolyl, benzoxazoyl benzimidazolyl, benzofuran, isobenzofuranyl, indolyl, isoindolyl, benzothiophenyl, isoquinolyl, quinoxalinyl, quinolyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituents for each of the above noted aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below. An “arylene” and a “heteroarylene,” alone or as part of another substituent, mean a divalent radical derived from an aryl and heteroaryl, respectively. A heteroaryl group substituent may be —O— bonded to a ring heteroatom nitrogen.
Spirocyclic rings are two or more rings wherein adjacent rings are attached through a single atom. The individual rings within spirocyclic rings may be identical or different.
Individual rings in spirocyclic rings may be substituted or unsubstituted and may have different substituents from other individual rings within a set of spirocyclic rings. Possible substituents for individual rings within spirocyclic rings are the possible substituents for the same ring when not part of spirocyclic rings (e.g. substituents for cycloalkyl or heterocycloalkyl rings). Spirocylic rings may be substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heterocycloalkylene and individual rings within a spirocyclic ring group may be any of the immediately previous list, including having all rings of one type (e.g. all rings being substituted heterocycloalkylene wherein each ring may be the same or different substituted heterocycloalkylene). When referring to a spirocyclic ring system, heterocyclic spirocyclic rings means a spirocyclic rings wherein at least one ring is a heterocyclic ring and wherein each ring may be a different ring. When referring to a spirocyclic ring system, substituted spirocyclic rings means that at least one ring is substituted and each substituent may optionally be different.
The symbol “” denotes the point of attachment of a chemical moiety to the remainder of a molecule or chemical formula.
The term “oxo,” as used herein, means an oxygen that is double bonded to a carbon atom.
The term “alkylarylene” as an arylene moiety covalently bonded to an alkylene moiety (also referred to herein as an alkylene linker). In embodiments, the alkylarylene group has the formula:
An alkylarylene moiety may be substituted (e.g. with a substituent group) on the alkylene moiety or the arylene linker (e.g. at carbons 2, 3, 4, or 6) with halogen, oxo, —N3, —CF3, —CCl3, —CBr3, —CI3, —CN, —CHO, —OH, —NH2, —COOH, —CONH2, —NO2, —SH, —SO2CH3 —SO3H, —OSO3H, —SO2NH2, —NHNH2, —ONH2, —NHC(O)NHNH2, substituted or unsubstituted C1-C5 alkyl or substituted or unsubstituted 2 to 5 membered heteroalkyl). In embodiments, the alkylarylene is unsubstituted.
Each of the above terms (e.g., “alkyl,” “heteroalkyl,” “cycloalkyl,” “heterocycloalkyl,” “aryl,” and “heteroaryl”) includes both substituted and unsubstituted forms of the indicated radical. Preferred substituents for each type of radical are provided below.
Substituents for the alkyl and heteroalkyl radicals (including those groups often referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) can be one or more of a variety of groups selected from, but not limited to, —OR′, ═O, ═NR′, ═N—OR′, —NR′R″, —SR′, -halogen, —SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO2R′, —CONR′R″, —OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)2R′, —NR—C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″, —S(O)R′, —S(O)2R′, —S(O)2NR′R″, —NRSO2R′, —NR′NR″R′″, —ONR′R″, —NR′C(O)NR″NR′″R″″, —CN, —NO2, —NR′SO2R″, —NR′C(O)R″, —NR′C(O)—OR″, —NR′OR″, in a number ranging from zero to (2m′+1), where m′ is the total number of carbon atoms in such radical. R, R′, R″, R′″, and R″″ each preferably independently refer to hydrogen, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl (e.g., aryl substituted with 1-3 halogens), substituted or unsubstituted heteroaryl, substituted or unsubstituted alkyl, alkoxy, or thioalkoxy groups, or arylalkyl groups. When a compound described herein includes more than one R group, for example, each of the R groups is independently selected as are each R′, R″, R′″, and R″″ group when more than one of these groups is present. When R′ and R″ are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a 4-, 5-, 6-, or 7-membered ring. For example, —NR′R″ includes, but is not limited to, 1-pyrrolidinyl and 4-morpholinyl. From the above discussion of substituents, one of skill in the art will understand that the term “alkyl” is meant to include groups including carbon atoms bound to groups other than hydrogen groups, such as haloalkyl (e.g., —CF3 and —CH2CF3) and acyl (e.g., —C(O)CH3, —C(O)CF3, —C(O)CH2OCH3, and the like).
Similar to the substituents described for the alkyl radical, substituents for the aryl and heteroaryl groups are varied and are selected from, for example: —OR′, —NR′R″, —SR′, -halogen, —SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO2R′, —CONR′R″, —OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)2R′, —NR—C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″, —S(O)R′, —S(O)2R′, —S(O)2NR′R″, —NRSO2R′, —NR′NR″R′″, —ONR′R″, —NR′C(O)NR″NR′″R″″, —CN, —NO2, —R′, —N3, —CH(Ph)2, fluoro(C1-C4)alkoxy, and fluoro(C1-C4)alkyl, —NR′SO2R″, —NR′C(O)R″, —NR′C(O)—OR″, —NR′OR″, in a number ranging from zero to the total number of open valences on the aromatic ring system; and where R′, R″, R′″, and R″″ are preferably independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl. When a compound described herein includes more than one R group, for example, each of the R groups is independently selected as are each R′, R″, R′″, and R″″ groups when more than one of these groups is present.
Substituents for rings (e.g. cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene) may be depicted as substituents on the ring rather than on a specific atom of a ring (commonly referred to as a floating substituent). In such a case, the substituent may be attached to any of the ring atoms (obeying the rules of chemical valency) and in the case of fused rings or spirocyclic rings, a substituent depicted as associated with one member of the fused rings or spirocyclic rings (a floating substituent on a single ring), may be a substituent on any of the fused rings or spirocyclic rings (a floating substituent on multiple rings). When a substituent is attached to a ring, but not a specific atom (a floating substituent), and a subscript for the substituent is an integer greater than one, the multiple substituents may be on the same atom, same ring, different atoms, different fused rings, different spirocyclic rings, and each substituent may optionally be different. Where a point of attachment of a ring to the remainder of a molecule is not limited to a single atom (a floating substituent), the attachment point may be any atom of the ring and in the case of a fused ring or spirocyclic ring, any atom of any of the fused rings or spirocyclic rings while obeying the rules of chemical valency. Where a ring, fused rings, or spirocyclic rings contain one or more ring heteroatoms and the ring, fused rings, or spirocyclic rings are shown with one more floating substituents (including, but not limited to, points of attachment to the remainder of the molecule), the floating substituents may be bonded to the heteroatoms. Where the ring heteroatoms are shown bound to one or more hydrogens (e.g. a ring nitrogen with two bonds to ring atoms and a third bond to a hydrogen) in the structure or formula with the floating substituent, when the heteroatom is bonded to the floating substituent, the substituent will be understood to replace the hydrogen, while obeying the rules of chemical valency.
Two or more substituents may optionally be joined to form aryl, heteroaryl, cycloalkyl, or heterocycloalkyl groups. Such so-called ring-forming substituents are typically, though not necessarily, found attached to a cyclic base structure. In one embodiment, the ring-forming substituents are attached to adjacent members of the base structure. For example, two ring-forming substituents attached to adjacent members of a cyclic base structure create a fused ring structure. In another embodiment, the ring-forming substituents are attached to a single member of the base structure. For example, two ring-forming substituents attached to a single member of a cyclic base structure create a spirocyclic structure. In yet another embodiment, the ring-forming substituents are attached to non-adjacent members of the base structure.
Two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally form a ring of the formula -T-C(O)—(CRR′)q—U—, wherein T and U are independently —NR—, —O—, —CRR′—, or a single bond, and q is an integer of from 0 to 3. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula -A-(CH2)r—B—, wherein A and B are independently —CRR′—, —O—, —NR—, —S—, —S(O)—, —S(O)2—, —S(O)2NR′—, or a single bond, and r is an integer of from 1 to 4. One of the single bonds of the new ring so formed may optionally be replaced with a double bond. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula —(CRR′)s—X′—(C″R″R′″)a—, where s and d are independently integers of from 0 to 3, and X′ is —O—, —NR′—, —S—, —S(O)—, —S(O)2—, or —S(O)2NR′—. The substituents R, R′, R″, and R′″ are preferably independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl.
As used herein, the terms “heteroatom” or “ring heteroatom” are meant to include oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), and silicon (Si).
A “substituent group,” as used herein, means a group selected from the following moieties:
A “size-limited substituent” or “size-limited substituent group,” as used herein, means a group selected from all of the substituents described above for a “substituent group,” wherein each substituted or unsubstituted alkyl is a substituted or unsubstituted C1-C20 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 20 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C8 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 8 membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted C6-C10 aryl, and each substituted or unsubstituted heteroaryl is a substituted or unsubstituted 5 to 10 membered heteroaryl.
A “lower substituent” or “lower substituent group,” as used herein, means a group selected from all of the substituents described above for a “substituent group,” wherein each substituted or unsubstituted alkyl is a substituted or unsubstituted C1-C8 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C7 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 7 membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted C6-C10 aryl, and each substituted or unsubstituted heteroaryl is a substituted or unsubstituted 5 to 9 membered heteroaryl.
In embodiments, a substituted or unsubstituted moiety (e.g., substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, and/or substituted or unsubstituted heteroarylene) is unsubstituted (e.g., is an unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl, unsubstituted heteroaryl, unsubstituted alkylene, unsubstituted heteroalkylene, unsubstituted cycloalkylene, unsubstituted heterocycloalkylene, unsubstituted arylene, and/or unsubstituted heteroarylene, respectively). In embodiments, a substituted or unsubstituted moiety (e.g., substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, and/or substituted or unsubstituted heteroarylene) is substituted (e.g., is a substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and/or substituted heteroarylene, respectively).
In embodiments, a substituted moiety (e.g., substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and/or substituted heteroarylene) is substituted with at least one substituent group, wherein if the substituted moiety is substituted with a plurality of substituent groups, each substituent group may optionally be different. In embodiments, if the substituted moiety is substituted with a plurality of substituent groups, each substituent group is different.
In embodiments, a substituted moiety (e.g., substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and/or substituted heteroarylene) is substituted with at least one size-limited substituent group, wherein if the substituted moiety is substituted with a plurality of size-limited substituent groups, each size-limited substituent group may optionally be different. In embodiments, if the substituted moiety is substituted with a plurality of size-limited substituent groups, each size-limited substituent group is different.
In embodiments, a substituted moiety (e.g., substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and/or substituted heteroarylene) is substituted with at least one lower substituent group, wherein if the substituted moiety is substituted with a plurality of lower substituent groups, each lower substituent group may optionally be different. In embodiments, if the substituted moiety is substituted with a plurality of lower substituent groups, each lower substituent group is different.
In embodiments, a substituted moiety (e.g., substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and/or substituted heteroarylene) is substituted with at least one substituent group, size-limited substituent group, or lower substituent group; wherein if the substituted moiety is substituted with a plurality of groups selected from substituent groups, size-limited substituent groups, and lower substituent groups; each substituent group, size-limited substituent group, and/or lower substituent group may optionally be different. In embodiments, if the substituted moiety is substituted with a plurality of groups selected from substituent groups, size-limited substituent groups, and lower substituent groups; each substituent group, size-limited substituent group, and/or lower substituent group is different.
In embodiments of the compounds herein, each substituted or unsubstituted alkyl may be a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted C1-C20 alkyl, each substituted or unsubstituted heteroalkyl is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted 2 to 20 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted C3-C8 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted 3 to 8 membered heterocycloalkyl, each or unsubstituted aryl is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted C6-C10 aryl, and/or each substituted or unsubstituted heteroaryl is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted 5 to 10 membered heteroaryl. In embodiments herein, each substituted or unsubstituted alkylene is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted C1-C20 alkylene, each substituted or unsubstituted heteroalkylene is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted 2 to 20 membered heteroalkylene, each substituted or unsubstituted cycloalkylene is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted C3-C8 cycloalkylene, each substituted or unsubstituted heterocycloalkylene is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted 3 to 8 membered heterocycloalkylene, each substituted or unsubstituted arylene is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted C6-C10 arylene, and/or each substituted or unsubstituted heteroarylene is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted 5 to 10 membered heteroarylene.
In embodiments, each substituted or unsubstituted alkyl is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted C1-C8 alkyl, each substituted or unsubstituted heteroalkyl is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted 2 to 8 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted C3-C7 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted 3 to 7 membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted C6-C10 aryl, and/or each substituted or unsubstituted heteroaryl is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted 5 to 9 membered heteroaryl. In embodiments, each substituted or unsubstituted alkylene is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted C1-C8 alkylene, each substituted or unsubstituted heteroalkylene is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted 2 to 8 membered heteroalkylene, each substituted or unsubstituted cycloalkylene is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted C3-C7 cycloalkylene, each substituted or unsubstituted heterocycloalkylene is a substituted or unsubstituted 3 to 7 membered heterocycloalkylene, each substituted or unsubstituted arylene is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted C6-C10 arylene, and/or each substituted or unsubstituted heteroarylene is a substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted 5 to 9 membered heteroarylene. In embodiments, the compound is a chemical species set forth in the Examples section, figures, or tables below.
Certain compounds provided herein possess asymmetric carbon atoms (optical or chiral centers) or double bonds; the enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisometric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino acids, and individual isomers are encompassed within the scope of the present disclosure. The compounds of provided herein do not include those that are known in art to be too unstable to synthesize and/or isolate. Compounds provided herein include those in racemic and optically pure forms. Optically active (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When the compounds described herein contain olefinic bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers.
As used herein, the term “isomers” refers to compounds having the same number and kind of atoms, and hence the same molecular weight, but differing in respect to the structural arrangement or configuration of the atoms.
The term “tautomer,” as used herein, refers to one of two or more structural isomers which exist in equilibrium and which are readily converted from one isomeric form to another.
It will be apparent to one skilled in the art that certain compounds provided herein may exist in tautomeric forms, all such tautomeric forms of the compounds being within the scope of the present disclosure.
Where the compounds disclosed herein have at least one chiral center, they may exist as individual enantiomers and diastereomers or as mixtures of such isomers, including racemates. Separation of the individual isomers or selective synthesis of the individual isomers is accomplished by application of various methods which are well known to practitioners in the art. Unless otherwise indicated, all such isomers and mixtures thereof are included in the scope of the compounds disclosed herein. Unless otherwise stated, structures depicted herein are also meant to include all stereochemical forms of the structure; i.e., the (R) and (S) configurations for each asymmetric center. Therefore, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the present compounds, generally recognized as stable by those skilled in the art, are within the scope of the present disclosure.
Unless otherwise stated, structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of a hydrogen by a deuterium or tritium, replacement of fluoride by 18F, or the replacement of a carbon by 13C- or 14C-enriched carbon are within the scope of the present disclosure.
The compounds provided herein may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may be radiolabeled with radioactive isotopes, such as for example tritium (3H), iodine-125 (125I), or carbon-14 (14C). All isotopic variations of the compounds provided herein, whether radioactive or not, are included within the present disclosure.
It should be noted that throughout the application that alternatives are written in Markush groups, for example, each amino acid position that contains more than one possible amino acid. It is specifically contemplated that each member of the Markush group should be considered separately, thereby comprising another embodiment, and the Markush group is not to be read as a single unit.
“Analog,” or “analogue” is used in accordance with its plain ordinary meaning within Chemistry and Biology and refers to a chemical compound that is structurally similar to another compound (i.e., a so-called “reference” compound) but differs in composition, e.g., in the replacement of one atom by an atom of a different element, or in the presence of a particular functional group, or the replacement of one functional group by another functional group, or the absolute stereochemistry of one or more chiral centers of the reference compound. Accordingly, an analog is a compound that is similar or comparable in function and appearance but not in structure or origin to a reference compound.
The terms “a” or “an,” as used in herein means one or more. In addition, the phrase “substituted with a[n],” as used herein, means the specified group may be substituted with one or more of any or all of the named substituents. For example, where a group, such as an alkyl or heteroaryl group, is “substituted with an unsubstituted C1-C20 alkyl, or unsubstituted 2 to 20 membered heteroalkyl,” the group may contain one or more unsubstituted C1-C20 alkyls, and/or one or more unsubstituted 2 to 20 membered heteroalkyls.
Where a moiety is substituted with an R substituent, the group may be referred to as “R-substituted.” Where a moiety is R-substituted, the moiety is substituted with at least one R substituent and each R substituent is optionally different. Where a particular R group is present in the description of a chemical genus (such as Formula (I)), a Roman decimal symbol may be used to distinguish each appearance of that particular R group. For example, where multiple R13 substituents are present, each R13 substituent may be distinguished as R13.1, R13.2, R13.3, R13.4, etc., wherein each of R13.1, R13.2, R13.3, R13.4, etc. is defined within the scope of the definition of R13 and optionally differently. The terms “a” or “an,” as used in herein means one or more. In addition, the phrase “substituted with a[n],” as used herein, means the specified group may be substituted with one or more of any or all of the named substituents. For example, where a group, such as an alkyl or heteroaryl group, is “substituted with an unsubstituted C1-C20 alkyl, or unsubstituted 2 to 20 membered heteroalkyl,” the group may contain one or more unsubstituted C1-C20 alkyls, and/or one or more unsubstituted 2 to 20 membered heteroalkyls.
Description of compounds of provided herein is limited by principles of chemical bonding known to those skilled in the art. Accordingly, where a group may be substituted by one or more of a number of substituents, such substitutions are selected so as to comply with principles of chemical bonding and to give compounds which are not inherently unstable and/or would be known to one of ordinary skill in the art as likely to be unstable under ambient conditions, such as aqueous, neutral, and several known physiological conditions. For example, a heterocycloalkyl or heteroaryl is attached to the remainder of the molecule via a ring heteroatom in compliance with principles of chemical bonding known to those skilled in the art thereby avoiding inherently unstable compounds.
Embodiments of the present disclosure relate to compounds targeted to the human peripheral myelin protein 22 (PMP22) mRNA (NCBI Reference Sequence NM_000304.4, deposited with GenBank on Nov. 22, 2018; SEQ ID NO: 1170). The compounds include double-stranded nucleic acids and single-stranded nucleic acids that act through the RNA interference pathway to inhibit the expression of the PMP22 mRNA. In embodiments, a compound is a double-stranded nucleic acid comprising an antisense strand complementary to the PMP22 mRNA and a sense strand complementary to the antisense strand. In embodiments, the antisense strand and sense strand of a compound are two separate strands and are not covalently linked and form a small interfering RNA (siRNA). In embodiments, the antisense strand and sense strand of a compound are covalently linked by a nucleotide linker to form a short hairpin RNA (shRNA). In embodiments, the compound is a single-stranded nucleic acid comprising an antisense strand complementary to the PMP22 mRNA (ssRNAi).
Provided herein are compounds comprising an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein each of the antisense strand and sense strands is 15 to 25 nucleotides in length, the nucleotide sequence of the antisense strand is at least 90% complementary to the human peripheral myelin protein 22 mRNA (SEQ ID NO: 1170), and the nucleotide sequence of the sense strand has no more than two mismatches to the nucleotide sequence of the antisense strand in the double-stranded region.
Provided herein are compounds comprising an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, each of the antisense strand and sense strands is 15 to 25 nucleotides in length, the nucleotide sequence of the antisense strand comprises at least 15 contiguous nucleotides of a nucleotide sequence selected from any one of SEQ ID NOs 491, 492, 493, 494, 495, 497, 498, 503, 504, 506, 510, 511, 514, 515, 516, 518, 524, 526, 529, 531, 532, 533, 534, 535, 536, 538, 539, 540, 541, 542, 543, 545, 546, 547, 548, 550, 553, 554, 556, 558, 559, 560, 561, 563, 567, 569, 575, 576, 579, 580, 581, 582, 583, 585, 590, 591, 595, 597, 600, 605, 609, 610, 618, 622, 623, 628, 630, 631, 633, 635, 637, 639, 641, 642, 643, 644, 645, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1122, 1124, 1125, 1126, 1127, 1128, 1129, 1130, 1131, 1132, 1133, 1134, 1135, 1136, 1137, 1138, 1139, 1140, 1141, 1142, 1143, 1144, 1145, 1146, 1147, 1148, 1149, 1150, 1151, 1152, 1153, 1154, 1155, 1156, 1157, 1158, 1159, 1160, 1161, 1162, 1163, 1164, 1165, 1166, 1167, 1168, 1169, 1118, 1121, 1123, 1126, and 1144, and the nucleotide sequence of the sense strand has no more than two mismatches to the nucleotide sequence of the antisense strand.
Provided herein are compounds comprising a single-stranded nucleic acid comprising an antisense strand, wherein the antisense strand is 15 to 25 nucleotides in length and the nucleotide sequence of the antisense strand comprises at least 15 contiguous nucleotides of a nucleotide sequence selected from any one of SEQ ID NOs 491, 492, 493, 494, 495, 497, 498, 503, 504, 506, 510, 511, 514, 515, 516, 518, 524, 526, 529, 531, 532, 533, 534, 535, 536, 538, 539, 540, 541, 542, 543, 545, 546, 547, 548, 550, 553, 554, 556, 558, 559, 560, 561, 563, 567, 569, 575, 576, 579, 580, 581, 582, 583, 585, 590, 591, 595, 597, 600, 605, 609, 610, 618, 622, 623, 628, 630, 631, 633, 635, 637, 639, 641, 642, 643, 644, 645, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1122, 1124, 1125, 1126, 1127, 1128, 1129, 1130, 1131, 1132, 1133, 1134, 1135, 1136, 1137, 1138, 1139, 1140, 1141, 1142, 1143, 1144, 1145, 1146, 1147, 1148, 1149, 1150, 1151, 1152, 1153, 1154, 1155, 1156, 1157, 1158, 1159, 1160, 1161, 1162, 1163, 1164, 1165, 1166, 1167, 1168, 1169, 1118, 1121, 1123, 1126, and 1144.
In embodiments, the nucleotide sequence of the antisense strand comprises at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, or 23 contiguous nucleotides selected from any one of SEQ ID NOs 491, 492, 493, 494, 495, 497, 498, 503, 504, 506, 510, 511, 514, 515, 516, 518, 524, 526, 529, 531, 532, 533, 534, 535, 536, 538, 539, 540, 541, 542, 543, 545, 546, 547, 548, 550, 553, 554, 556, 558, 559, 560, 561, 563, 567, 569, 575, 576, 579, 580, 581, 582, 583, 585, 590, 591, 595, 597, 600, 605, 609, 610, 618, 622, 623, 628, 630, 631, 633, 635, 637, 639, 641, 642, 643, 644, 645, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1122, 1124, 1125, 1126, 1127, 1128, 1129, 1130, 1131, 1132, 1133, 1134, 1135, 1136, 1137, 1138, 1139, 1140, 1141, 1142, 1143, 1144, 1145, 1146, 1147, 1148, 1149, 1150, 1151, 1152, 1153, 1154, 1155, 1156, 1157, 1158, 1159, 1160, 1161, 1162, 1163, 1164, 1165, 1166, 1167, 1168, 1169, 1118, 1121, 1123, 1126, and 1144.
In embodiments, the nucleotide sequence of the antisense strand comprises 19 contiguous nucleotides of a nucleotide sequence selected from any one of SEQ ID NOs 491, 492, 493, 494, 495, 497, 498, 503, 504, 506, 510, 511, 514, 515, 516, 518, 524, 526, 529, 531, 532, 533, 534, 535, 536, 538, 539, 540, 541, 542, 543, 545, 546, 547, 548, 550, 553, 554, 556, 558, 559, 560, 561, 563, 567, 569, 575, 576, 579, 580, 581, 582, 583, 585, 590, 591, 595, 597, 600, 605, 609, 610, 618, 622, 623, 628, 630, 631, 633, 635, 637, 639, 641, 642, 643, 644, 645, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1122, 1124, 1125, 1126, 1127, 1128, 1129, 1130, 1131, 1132, 1133, 1134, 1135, 1136, 1137, 1138, 1139, 1140, 1141, 1142, 1143, 1144, 1145, 1146, 1147, 1148, 1149, 1150, 1151, 1152, 1153, 1154, 1155, 1156, 1157, 1158, 1159, 1160, 1161, 1162, 1163, 1164, 1165, 1166, 1167, 1168, 1169, 1118, 1121, 1123, 1126, and 1144.
Provided below are features of compounds, such as length, nucleotide sequence, and nucleotide modifications. It is understood that an embodiment of an antisense strand may apply to the antisense strand of a single-stranded nucleic acid or a double-stranded nucleic acid. Further, it is understood that an embodiment of a sense strand may apply to a sense strand of any double-stranded nucleic acid provided herein, including siRNAs and shRNAs.
In embodiments, an antisense strand is 15 to 25 nucleotides in length. In embodiments, an antisense strand is 17 to 23 nucleotides in length. In embodiments, an antisense strand is 19 to 21 nucleotides in length. In embodiments, an antisense strand is 21 to 23 nucleotides in length. In embodiments, an antisense strand is 15 nucleotides in length. In embodiments, an antisense strand is 16 nucleotides in length. In embodiments, an antisense strand is 17 nucleotides in length. In embodiments, an antisense strand is 18 nucleotides in length. In embodiments, an antisense strand is 19 nucleotides in length. In embodiments, an antisense strand is 20 nucleotides in length. In embodiments, an antisense strand is 21 nucleotides in length. In embodiments, an antisense strand is 22 nucleotides in length. In embodiments, an antisense strand is 23 nucleotides in length. In embodiments, an antisense strand is 24 nucleotides in length. In embodiments, an antisense strand is 25 nucleotides in length.
In embodiments, the nucleotide sequence of the antisense strand is at least 95% complementary to SEQ ID NO: 1170. In embodiments, the nucleotide sequence of the antisense strand is 100% complementary to SEQ ID NO: 1170. In embodiments, the nucleotide sequence of the antisense strand is 100% complementary to nucleotides 213 to 233 of SEQ ID NO: 1170.
In embodiments, a sense strand is 15 to 25 nucleotides in length. In embodiments, a sense strand is 17 to 23 nucleotides in length. In embodiments, a sense strand is 19 to 21 nucleotides in length. In embodiments, a sense strand is 21 to 23 nucleotides in length. In embodiments, a sense strand is 15 nucleotides in length. In embodiments, a sense strand is 16 nucleotides in length. In embodiments, a sense strand is 17 nucleotides in length. In embodiments, a sense strand is 18 nucleotides in length. In embodiments, a sense strand is 19 nucleotides in length. In embodiments, a sense strand is 20 nucleotides in length. In embodiments, a sense strand is 21 nucleotides in length. In embodiments, a sense strand is 22 nucleotides in length. In embodiments, a sense strand is 23 nucleotides in length. In embodiments, a sense strand is 24 nucleotides in length. In embodiments, a sense strand is 25 nucleotides in length.
In embodiments, length of the sense strand is identical to the length of the antisense strand. In embodiments, the length of the sense strand is greater than the length of the antisense strand. In embodiments, the length of the sense strand is less than the length of the antisense strand.
The double-stranded region of a double-stranded nucleic acid may be from 15 to 25 nucleobase pairs in length, depending on the lengths of the sense strand and the antisense strand. In embodiments, the double-stranded region is 17 to 23 nucleobase pairs in length. In embodiments, the double-stranded region is 19 to 21 nucleobase pairs in length. In embodiments, the double-stranded region is 21 to 23 nucleotides in length. In embodiments, the double-stranded region is 15 nucleobase pairs in length. In embodiments, the double-stranded region is 16 nucleobase pairs in length. In embodiments, the double-stranded region is 17 nucleobase pairs in length. In embodiments, the double-stranded region is 18 nucleobase pairs in length. In embodiments, the double-stranded region is 19 nucleobase pairs in length. In embodiments, the double-stranded region is 20 nucleobase pairs in length.
In embodiments, the double-stranded region is 21 nucleobase pairs in length. In embodiments, the double-stranded region is 22 nucleobase pairs in length. In embodiments, the double-stranded region is 23 nucleobase pairs in length. In embodiments, the double-stranded region is 24 nucleobase pairs in length. In embodiments, the double-stranded region is 25 nucleobase pairs in length.
In embodiments, the nucleotide sequence of a sense strand has no more than one mismatch to the nucleotide sequence of an antisense strand of a double-stranded nucleic acid. In embodiments, the nucleotide sequence of a sense strand has no mismatches to the nucleotide sequence of an antisense strand of a double-stranded nucleic acid. Single-stranded nucleotide overhangs and nucleotide linkers are not considered for the purposes of determining the number of mismatches within the double-stranded region of a double-stranded nucleic acid provided herein. For example, a double-stranded nucleic acid comprising an antisense strand that is 23 nucleotides in length, and a sense strand that is 21 nucleotides in length have no mismatches over the double-stranded region, provided the nucleotide sequence of the sense strand is fully complementary over its length the nucleotide sequence of the antisense strand. Alternatively, a double-stranded nucleic acid comprising a sense strand that is 20 nucleotides in length, an antisense strand that is 22 nucleotides in length, and a nucleotide linker that is eight nucleotides in length, may have no mismatches over the double-stranded region provided the nucleotide sequence of the sense strand is fully complementary over its length to the nucleotide sequence of the antisense strand.
In embodiments, a double-stranded nucleic acid comprises an antisense strand of 19 nucleotides in length and a sense strand of 19 nucleotides in length. In embodiments, the antisense strand is 22 nucleotides in length and the sense strand is 20 nucleotides in length. In embodiments, the antisense strand is 23 nucleotides in length and the sense strand is 21 nucleotides in length. In embodiments, the antisense strand is 23 nucleotides in length including two deoxythymidines at the 3′ terminus, and the sense strand is 21 nucleotides in length including two deoxythymidines at the 3′ terminus.
In embodiments of compound comprising double-stranded nucleic acid where the antisense strand and sense strand are separate strands that are not covalently linked, the terminal nucleotides may form a nucleobase pair, in which case the end of the double-stranded nucleic acid is a blunt end. Alternatively, one or more unpaired nucleotides of an antisense strand and/or sense strand may extend beyond the terminus of the complementary strand, resulting in a nucleotide overhang of one or more terminal single-stranded nucleotides. In embodiments, at least one of the 5′ and 3′ terminus of a double-stranded nucleic acid is a blunt end. In embodiments, both the 5′ terminus and 3′ terminus of the double-stranded nucleic acid are blunt ends. In embodiments, at least one end of the double-stranded nucleic acid comprises a nucleotide overhang. In embodiments, each end of the double-stranded nucleic acid comprises a nucleotide overhang. In embodiments, one end of the double-stranded nucleic acid is a blunt end and the other end of the double-stranded nucleic acid comprises a nucleotide overhang. In embodiments, the antisense strand comprises a nucleotide overhang at its 3′ terminus. In embodiments, the sense strand comprises a nucleotide overhang at its 3′ terminus. In embodiments, each of the antisense strand and sense strand comprises a nucleotide overhang at its 3′ terminus. In embodiments, at least one of the antisense strand and sense strand comprises a nucleotide overhang at its 5′ terminus. In embodiments, each of the antisense strand and sense strand comprises a nucleotide overhang at each 5′ terminus.
In embodiments, a nucleotide overhang is from one to five single-stranded nucleotides. In embodiments, a nucleotide overhang is one single-stranded nucleotide. In embodiments, a nucleotide overhang is two single-stranded nucleotides. In embodiments, a nucleotide overhang is three single-stranded nucleotides. In embodiments, a nucleotide overhang is three single-stranded nucleotides. In embodiments, a nucleotide overhang is four single-stranded nucleotides. In embodiments, a nucleotide overhang is five single-stranded nucleotides. In embodiments, at least one of the single-stranded nucleotides of a nucleotide overhang is a modified nucleotide. In embodiments, each of the single-stranded nucleotides of a nucleotide overhang is a modified nucleotide. In embodiments, the modified nucleotide is a 2′-O-methyl nucleotide. In embodiments, the nucleotide overhang is two single-stranded nucleotides and each nucleotide is a 2′-O-methoxyethyl nucleotide.
In embodiments, at least one nucleotide of the nucleotide overhang at the 3′ terminus of an antisense strand is complementary to a corresponding nucleotide of SEQ ID NO: 1170. In embodiments, each nucleotide of the nucleotide overhang at the 3′ terminus of an antisense strand is complementary to a corresponding nucleotide of SEQ ID NO: 1170. In some embodiment, at least one nucleotide of the nucleotide overhang at the 3′ terminus of an antisense strand is not complementary to a corresponding nucleotide of SEQ ID NO: 1170. In embodiments, each nucleotide of the nucleotide overhang at the 3′ terminus of an antisense strand is not complementary to a corresponding nucleotide of SEQ ID NO: 1170.
In embodiments, at least one single-stranded nucleotide of a nucleotide overhang is a deoxythymidine nucleotide. In embodiments, a nucleotide overhang is two single-stranded nucleotides and each nucleotide is a deoxythymidine nucleotide. In embodiments, the nucleotide sequence of the antisense strand comprises a nucleotide overhang of two deoxythymidine nucleotides. In embodiments, the sense strand comprises a nucleotide overhang of two deoxythymidine nucleotides. In embodiments, the antisense strand and the sense strand comprise a nucleotide overhang of two deoxythymidine nucleotides.
Non-limiting examples of double-stranded nucleic acids comprising blunt ends or nucleotide overhangs are provided in Table 1 below.
In the first example, where the antisense strand is 21 nucleotides in length and the sense strand is 21 nucleotides in length, and the nucleotide sequence of the antisense strand is fully complementary to the nucleotide sequence of the sense strand over the double-stranded region, the length of the double-stranded region is 19 nucleobase pairs and each terminus of the double-stranded nucleic acid has a dTdT overhang.
In the second example, where the antisense strand is 21 nucleotides in length and the sense strand is 19 nucleotides in length, and the nucleotide sequence of the antisense strand is fully complementary to the nucleotide sequence of the sense strand over the double-stranded region, the length of the double-stranded region is 19 nucleobase pairs and the 3′ terminus of the antisense strand comprises a dTdT overhang.
In the third example, where the antisense strand is 19 nucleotides in length and the sense strand is 19 nucleotides in length, and the nucleotide sequence of the antisense strand is fully complementary to the nucleotide sequence of the sense strand over the double-stranded region, the length of the double-stranded region is 19 nucleobase pairs and each terminus is a blunt end.
In the fourth example, where the antisense strand is 23 nucleotides in length and the sense strand is 21 nucleotides in length, the length of the double-stranded region is 21 nucleobase pairs and 3′ terminus of the antisense strand comprises a two-nucleotide overhang.
In embodiments of a double-stranded nucleic acid comprising a nucleotide linker, the termini that are not connected by the nucleotide linker may form a blunt end or may form a nucleotide overhang of one or more single-stranded nucleotides. In embodiments, the non-linked end of the double-stranded nucleic acid is a blunt end. In embodiments, the non-linked end comprises a nucleotide overhang of one or more single-stranded nucleotides.
In embodiments, the non-linked end of the guide strand comprises a nucleotide overhang. In embodiments, the non-linked end of the sense strand comprises a nucleotide overhang. In embodiments, the 3′ terminus of the guide strand comprises a nucleotide overhang. In embodiments, the 3′ terminus of the sense strand comprises a nucleotide overhang. In embodiments, the 5′ terminus of the sense strand comprises a nucleotide overhang. In embodiments, the 5′ terminus of the sense strand comprises a nucleotide overhang.
In embodiments of a double-stranded nucleic acid where the antisense and sense strand are covalently linked by a nucleotide linker, the nucleotide linker is four to 16 nucleotides in length. In embodiments, the nucleotide linker is four nucleotides in length. In embodiments, the nucleotide linker is four nucleotides in length. In embodiments, the nucleotide linker is five nucleotides in length. In embodiments, the nucleotide linker is six nucleotides in length. In embodiments, the nucleotide linker is seven nucleotides in length. In embodiments, the nucleotide linker is eight nucleotides in length. In embodiments, the nucleotide linker is nine nucleotides in length. In embodiments, the nucleotide linker is 10 nucleotides in length. In embodiments, the nucleotide linker is 11 nucleotides in length. In embodiments, the nucleotide linker is 12 nucleotides in length. In embodiments, the nucleotide linker is 13 nucleotides in length. In embodiments, the nucleotide linker is 14 nucleotides in length. In embodiments, the nucleotide linker is 15 nucleotides in length. In embodiments, the nucleotide linker is 16 nucleotides in length.
Although the sequence listing accompanying this filing identifies each nucleotide sequence as either “RNA” or “DNA” as required, in practice, those sequences may be modified with a combination of chemical modifications specified herein. One of skill in the art will readily appreciate that in the sequence listing, such designation as “RNA” or “DNA” to describe modified nucleotides is somewhat arbitrary. For example, a nucleic acid provided herein comprising a nucleotide comprising a 2′-O-methyl sugar moiety and a thymine base may be described as a DNA residue in the sequence listing, even though the nucleotide is modified and is not a naturally-occurring DNA nucleotide.
Accordingly, nucleic acid sequences provided 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, a nucleic acid having the nucleotide sequence “ATCGATCG” in the sequence listing encompasses any nucleic acid having such nucleotide sequence, whether modified or unmodified, including, but not limited to, such nucleic acids comprising RNA bases, such as those having sequence “AUCGAUCG” and those having some DNA bases and some RNA bases such as “AUCGATCG” and oligonucleotides having other modified bases, such as “ATmeCGAUCG,” wherein meC indicates a 5-methylcytosine.
Double-stranded and single-stranded nucleic acids provided herein may comprise one or more modified nucleotides. A modified nucleotide may be selected over an unmodified form because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for other oligonucleotides or nucleic acid targets, increased stability in the presence of nucleases, and/or reduced immune stimulation.
In embodiments, at least one nucleotide of the antisense strand is a modified nucleotide. In embodiments, at least one nucleotide of the sense strand is a modified nucleotide. In embodiments, each nucleotide of the antisense strand forming the double-stranded region is a modified nucleotide. In embodiments, each nucleotide of the sense strand forming the double-stranded region comprises is a modified nucleotide.
In embodiments, a modified nucleotide comprises one or more of a modified sugar moiety, a modified internucleotide linkage, and a 5′-terminal modified phosphate group. In embodiments, a modified nucleotide comprises a modified sugar moiety. In embodiments, a modified nucleotide comprises a modified internucleotide linkage. In embodiments, a modified nucleotide comprises a modified nucleobase. In embodiments, a modified nucleotide comprises a modified 5′-terminal phosphate group. In embodiments, a modified nucleotide comprises a modification at the 5′ carbon of the pentafuranosyl sugar. In embodiments, a modified nucleotide comprises a modification at the 3′ carbon of the pentafuranosyl sugar. In embodiments, a modified nucleotide comprises a modification at the 2′ carbon of the pentafuranosyl sugar. In embodiments, a modified nucleotide is at the 5′ terminus of an antisense strand or sense strand. In embodiments, a modified nucleotide is at the 3′ terminus of an antisense strand or sense strand. In embodiments, a modified nucleotide is at an internal nucleotide of an antisense strand or sense strand. In embodiments, a modified nucleotide comprises a ligand attached to the 2′, 3, or 5′ carbon of the pentafuranosyl sugar.
In embodiments, a nucleotide comprises a ligand attached to a nucleobase.
A modified nucleotide may comprise a modified sugar moiety, a naturally occurring nucleobase, and a naturally occurring internucleotide linkage. A modified nucleotide may comprise a modified sugar moiety, a naturally occurring nucleobase, and a modified internucleotide linkage.
In embodiments, a modified sugar moiety is modified at the 2′ carbon of the pentafuranosyl sugar, relative to the naturally occurring 2′-OH of RNA or the 2′-H of DNA.
In embodiments, a modification at the 2′ carbon of the pentafuranosyl sugar is selected from F, OCF3, OCH3 (also referred to as “2′-OMe” or “2′-O-methyl), OCH2CH2OCH3 (also referred to as “2′-O-methoxyethyl” or “2′-MOE”), 2′-O(CH2)2SCH3, O—(CH2)2—O—N(CH3)2, —O(CH2)2O(CH2)2N(CH3)2, and O—CH2—C(═O)—N(H)CH3.
In embodiments, a modified sugar moiety is a 2′-fluoro sugar (also referred to as a 2′-F sugar). In embodiments, a modified sugar moiety is a 2′-O-methyl sugar (also referred to as a “2′-OMe sugar” or a “2′-OCH3” sugar). In embodiments, a modified sugar moiety is a 2′-O-methoxyethyl sugar (also referred to as a 2′-OCH2CH2OCH3 or a 2′-MOE sugar).
In embodiments, the modified nucleotide comprising a modified sugar moiety is selected from a 2′-fluoro nucleotide, a 2′-O-methyl nucleotide, a 2′-O-methoxyethyl nucleotide, and a bicyclic sugar nucleotide. In embodiments, a modified nucleotide is a 2′-fluoro nucleotide, where the 2′ carbon of the pentafuranosyl sugar has a fluoro substitution. In embodiments, a modified nucleotide is a 2′-O-methyl nucleotide, where the 2′ carbon of the pentafuranosyl sugar has a 2′-O methyl substitution. In embodiments, a modified nucleotide is a 2′-O-methoxyethyl nucleotide, where the 2′ carbon of the pentafuranosyl sugar has a 2′-O-methoxyethyl substitution. Other modified nucleotides may be similarly named.
In embodiments, a modified nucleotide comprises a modified sugar moiety, where the ribose has a covalent linkage between the 2′ and 4′ carbons. Such a modified sugar moiety may be referred to as a “bicyclic sugar,” and nucleotides comprising such sugar moieties may be referred to as “bicyclic nucleic acids.” In embodiments, the covalent linkage of a bicyclic sugar is a methyleneoxy linkage (4′-CH2—O-2′), also known as “LNA.” In embodiments, the covalent linkage of a bicyclic sugar is an ethyleneoxy linkage (4′-(CH2)2—O-2′), also known as “ENA.” In embodiments, the covalent linkage of a bicyclic moiety is a methyl(methyleneoxy) linkage (4′-CH(CH3)—O-2′), also known as “constrained ethyl” or “cEt.” In certain embodiments, the —CH(CH3)— bridge is constrained in the S orientation (“S-cEt”). In certain embodiments, the —CH(CH3)— bridge is constrained in the R orientation (“R-cEt”). In embodiments, the covalent linkage of a bicyclic sugar is a (4′-CH(CH2—OMe)-O-2′ linkage, also known as “c-MOE.” In embodiments, the bicyclic sugar is a D sugar in the alpha configuration. In certain such embodiments, the bicyclic sugar is a D sugar in the beta configuration. In certain such embodiments, the bicyclic sugar is an L sugar in the alpha configuration. In certain such embodiments, the bicyclic sugar is an L sugar in the beta configuration.
In embodiments, a modified sugar moiety is a 1,5-anhydrohexitol nucleic acid, also known as a “hexitol nucleic acid” or “HNA.”
In embodiments, the oxygen of the pentafuranosyl sugar is replace with a sulfur, to form a thio-sugar. In embodiments, a thio-sugar is modified at the 2′ carbon.
In embodiments, a modified internucleotide linkage is a phosphorothioate internucleotide linkage. In embodiments, a modified internucleotide linkage is a methylphosphonate internucleotide linkage.
In embodiments, the first two internucleotide linkages at the 5′ terminus of the sense strand and the last two internucleotide linkages at the 3′ terminus of the sense strand are phosphorothioate internucleotide linkages. In embodiments, the first two internucleotide linkages at the 5′ terminus of the antisense strand and the last two internucleotide linkages at the 3′ terminus of the antisense strand are phosphorothioate internucleotide linkages. In embodiments, the first two internucleotide linkages at the 5′ terminus of the sense strand and the last two internucleotide linkages at the 3′ terminus of the sense strand are phosphorothioate internucleotide linkages, and the first two internucleotide linkages at the 5′ terminus of the antisense strand and the last two internucleotide linkages at the 3′ terminus of the antisense strand are phosphorothioate internucleotide linkages.
In embodiments, a modified nucleobase is selected from 5-hydroxymethyl cytosine, 7-deazaguanine and 7-deazaadenine. In embodiments, a modified nucleobase is selected from 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine and 2-pyridone. In embodiments, a modified nucleobase is selected from 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and 0-6 substituted purines, including 2 aminopropyladenine, 5-propynyluracil and 5-propynylcytosine.
In embodiments, a modified nucleotide comprises a modification of the phosphate group at the 5′-carbon of the pentafuranosyl sugar. In embodiments, the modified phosphate group is 5′-(E)-vinylphosphonate (5′-VP).
In embodiments, a modified nucleotide is a phosphorodiamidite-linked morpholino nucleotide.
In embodiments, a modified nucleotide comprises an acyclic nucleoside derivative lacking the bond between the 2′ carbon and 3′ carbon of the sugar ring, also known as an “unlocked nucleic acid” or “UNA.”
In embodiments, the antisense strand is 21 nucleotides in length and the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, and 19 are 2′-O-methyl nucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, and 18 are 2′-fluoro nucleotides, and nucleotides 20 and 21 are beta-D-deoxynucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the following Pattern I:
5′-NMSNFSNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMSNSN-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, “N” is a beta-D-deoxynucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, and 19 are 2′-fluoro nucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, and 18 are 2′-O-methyl nucleotides, and nucleotides 20 and 21 are beta-D-deoxynucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the following Pattern II:
5′-NFSNMSNFNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNFSNSN-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, “N” is a beta-D-deoxynucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the antisense strand is 19 nucleotides in length and the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, and 19 are 2′-O-methyl nucleotides and nucleotides 2, 4, 6, 8, 10, 12, 14, 16, and 18 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the following Pattern III:
5′-NMSNFSNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMSNFSNM-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, “N” is a beta-D-deoxynucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkages is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 19 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, and 19 are 2′-fluoro nucleotides and nucleotides 2, 4, 6, 8, 10, 12, 14, 16, and 18 are 2′-O-methyl nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the following Pattern IV:
5′-NFSNMSNFNMNFNMNFNMNFNMNFNMNFNMNFNMNFSNMSNF-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-flouro nucleotide, “N” is a beta-D-deoxynucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphorodiester internucleotide linkage.
In embodiments, the antisense strand is 23 nucleotides in length and the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides and nucleotides 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 are 2′-fluoro nucleotides the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the following Pattern V:
5′-NMSNFSNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMSNMSNM-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, wherein the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 21 are 2′-fluoro nucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 are 2′-O-methyl nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern VI:
5′-NFSNMSNFNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNFSNMSNF-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the antisense strand is 23 nucleotides in length and the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 12, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides and nucleotides 2, 4, 6, 8, 10, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern VII:
5′-NMSNFSNMNFNMNFNMNFNMNFNMNMNMNFNMNFNMNFNMNFNMSNMSNM-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 10, 11, 13, 15, 17, 19, and 21 are 2′-fluoronucleotides, nucleotides 2, 4, 6, 8, 12, 14, 16, 18, and 20 are 2′-O-methyl nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern VIII:
5′-NFSNMSNFNMNFNMNFNMNFNFNFNMNFNMNFNMNFNMNFSNMSNF-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the antisense strand is 23 nucleotides in length and the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 10, 11, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides and nucleotides 2, 4, 6, 8, 12, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern IX:
5′-NMSNFSNMNFNMNFNMNFNMNMNMNFNMNFNMNFNMNFNMNFNMSNMSNM-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 11, 12, 13, 15, 17, 19, and 21 are 2′-fluoronucleotides, nucleotides 2, 4, 6, 8, 10, 14, 16, 18, and 20 are 2′-O-methyl nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern X:
5′-NFSNMSNFNMNFNMNFNMNFNMNFNFNFNMNFNMNFNMNFSNMSNF-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 23 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 21 are 2′-fluoronucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 are 2′-O-methyl nucleotides, nucleotides 22 and 23 are beta-D-deoxynucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern XI:
5′-NFSNMSNFNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNFSNSN-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, “N” is a beta-D-deoxynucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 23 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 10, 11, 13, 15, 17, 19, and 21 are 2′-fluoronucleotides, nucleotides 2, 4, 6, 8, 12, 14, 16, 18, and 20 are 2′-O-methyl nucleotides, nucleotides 22 and 23 are beta-D-deoxynucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern XII:
5′-NFSNMSNFNMNFNMNFNMNFNFNFNMNFNMNFNMNFNMNFNMNFSNSN-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, “N” is a beta-D-deoxynucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 23 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 11, 12, 13, 15, 17, 19, and 21 are 2′-fluoronucleotides, nucleotides 2, 4, 6, 8, 10, 14, 16, 18, and 20 are 2′-O-methyl nucleotides, nucleotides 22 and 23 are beta-D-deoxynucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern XIII:
5′-NFSNMSNFNMNFNMNFNMNFNMNFNFNFNMNFNMNFNMNFNMNFSNSN-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, “N” is a beta-D-deoxynucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 2, 3, 4, 6, 8, 12, 14, 16, 18, 19, 20, and 21 are 2′-methyl nucleotides, nucleotides 5, 7, 9, 10, 11, 13, 15, and 17 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern XIV:
5′-NMSNMSNMNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNMSNMSNM-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 2, 3, 4, 6, 8, 12, 14, 16, 18, 19, 20, and 21 are 2′-O-methyl nucleotides, nucleotides 5, 7, 9, 10, 11, 13, 15, and 17 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern XV:
5′-NMSNMSNMNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNMSNMSNM-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the antisense strand is 23 nucleotides in length and the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 8, 9, 11, 12, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides, nucleotides 2, 4, 6, 10, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern XVI:
5′-NMSNFSNMNFNMNFNMNFNMNMNMNFNMNFNMNFNMNFNMNFNMSNMSNM-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, “N” is a beta-D-deoxynucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the antisense strand is 23 nucleotides in length and the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 15, 17, 18, 19, 20, 21, 22, and 23 are 2′-O-methyl nucleotides, nucleotides 2, 6, 14, and 16 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern XVII:
5′-NMSNFSNMNFNMNFNMNFNMNMNMNFNMNFNMNFNMNFNMNFNMSNMSNM-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, “N” is a beta-D-deoxynucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21 are 2′-O-methyl nucleotides, nucleotides 7, 9, 10, and 11 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern XVIII:
5′-NMSNMSNMNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNMSNMSNM-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21 are 2′-O-methyl nucleotides, nucleotides 7, 9, 10, and 11 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern XIX:
5′-NMSNMSNMNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNMSNMSNM-3′, wherein “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1 and 2 are 2′-O-methoxyethyl nucleotides, nucleotides 3, 4, 6, 8, 12, 14, 16, 18, 19, 20, and 21 are 2′-O-methyl nucleotides, nucleotides 5, 7, 9, 10, 11, 13, 15, and 17 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern XX:
5′-NESNESNMNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNMSNMSNM-3′, wherein “NE” is a 2′-O-methoxyethyl nucleotide, “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 2 and 3 are 2′-O-methoxyethyl nucleotides, nucleotides 1, 4, 6, 8, 12, 14, 16, 18, 19, 20, and 21 are 2′-O-methyl nucleotides, nucleotides 5, 7, 9, 10, 11, 13, 15, and 17 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern XXI:
5′-NESNESNMNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNMSNMSNM-3′, wherein “NE” is a 2′-O-methoxyethyl nucleotide, “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 2, 3, 19 and 20 are 2′-O-methoxyethyl nucleotides, nucleotides 1, 4, 6, 8, 12, 14, 16, 18, and 21 are 2′-O-methyl nucleotides, nucleotides 5, 7, 9, 10, 11, 13, 15, and 17 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern XXII:
5′-NESNESNMNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNMSNMSNM-3′, wherein “NE” is a 2′-O-methoxyethyl nucleotide, “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 2, 3, and 4 are 2′-O-methoxyethyl nucleotides, nucleotides 6, 8, 12, 14, 16, 18, 19, 20 and 21 are 2′-O-methyl nucleotides, nucleotides 5, 7, 9, 10, 11, 13, 15, and 17 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. Such a modification pattern may be represented by the Pattern XXIII:
5′-NESNESNMNMNFNMNFNMNFNMNFNMNFNMNFNMNFNMNMSNMSNM-3′, wherein “NE” is a 2′-O-methoxyethyl nucleotide, “NM” is a 2′-O-methyl nucleotide, “NF” is a 2′-fluoro nucleotide, a superscript “S” is a phosphorothioate internucleotide linkage, and each other internucleotide linkage is a phosphodiester internucleotide linkage.
In embodiments, an antisense strand has the modification pattern of Pattern I and a 5′-VP at the 5′-terminal nucleotide. In embodiments, an antisense strand has the modification pattern of Pattern III and a 5′-VP at the 5′-terminal nucleotide. In embodiments, an antisense strand has the modification pattern of Pattern V and a 5′-VP at the 5′-terminal nucleotide. In embodiments, an antisense strand has the modification pattern of Pattern VII and a 5′-VP at the 5′ terminal nucleotide. In embodiments, an antisense strand has the modification pattern of Pattern IX and a 5′-VP at the 5′ terminal nucleotide. In embodiments, an antisense strand has the modification pattern of Pattern XVI and a 5′-VP at the 5′ terminal nucleotide. In embodiments, an antisense strand has the modification pattern of Pattern XVII and a 5′-VP at the 5′ terminal nucleotide.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded region, wherein the antisense strand and sense strand are not covalently linked (i.e. the antisense strand and sense strand form an siRNA), wherein the antisense strand is 21 nucleotides in length and the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, and 19 are 2′-O-methyl nucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, and 18 are 2′-fluoro nucleotides, and nucleotides 20 and 21 are beta-D-deoxynucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, and 19 are 2′-fluoro nucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, and 18 are 2′-O-methyl nucleotides, and nucleotides 20 and 21 are beta-D-deoxynucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern represented by Pattern I and the sense strand has the modification pattern represented by Pattern II.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and sense strand are not covalently linked (i.e. the antisense strand and sense strand form an siRNA), wherein the antisense strand is 19 nucleotides in length and the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, and 19 are 2′-O-methyl nucleotides and nucleotides 2, 4, 6, 8, 10, 12, 14, 16, and 18 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 21 nucleotide in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, and 19 are 2′-fluoro nucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, and 18 are 2′-O-methyl nucleotides, and nucleotides 20 and 21 are beta-D-deoxynucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern represented by Pattern III and the sense strand has the modification pattern represented by Pattern II.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and sense strand are not covalently linked (i.e. the antisense strand and sense strand form an siRNA), wherein the antisense strand is 21 nucleotides in length and the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, and 19 are 2′-O-methyl nucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, and 18 are 2′-fluoro nucleotides, and nucleotides 20 and 21 are beta-D-deoxy nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 19 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, and 19 are 2′-fluoro nucleotides and nucleotides 2, 4, 6, 8, 10, 12, 14, 16, and 18 are 2′-O-methyl nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern represented by Pattern I and the sense strand has the modification pattern represented by Pattern IV.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and sense strand are not covalently linked (i.e. the antisense strand and sense strand form an siRNA), wherein the antisense strand is 19 nucleotides in length and the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, and 19 are 2′-O-methyl nucleotides and nucleotides 2, 4, 6, 8, 10, 12, 14, 16, and 18 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide is a phosphodiester internucleotide linkage; and wherein the sense strand is 19 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, and 19 are 2′-fluoro nucleotides and nucleotides 2, 4, 6, 8, 10, 12, 14, 16, and 18 are 2′-O-methyl nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern represented by Pattern III and the sense strand has the modification pattern represented by Pattern IV.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and sense strand are not covalently linked (i.e. the antisense strand and sense strand form an siRNA), wherein the antisense strand is 23 nucleotides in length and wherein the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides and nucleotides 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 are 2′-fluoro nucleotides the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 21 are 2′-fluoro nucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 are 2′-O-methyl nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern represented by Pattern V and the sense strand has the modification represented by Pattern VI.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and sense strand are not covalently linked (i.e. the antisense strand and sense strand form an siRNA), wherein the antisense strand is 23 nucleotides in length and wherein the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 12, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides and nucleotides 2, 4, 6, 8, 10, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 10, 11, 13, 15, 17, 19, and 21 are 2′-fluoronucleotides, nucleotides 2, 4, 6, 8, 12, 14, 16, 18, and 20 are 2′-O-methyl nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern represented by Pattern VII and the sense strand has the modification pattern represented by Pattern VIII.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and sense strand are not covalently linked (i.e. the antisense strand and sense strand form an siRNA), wherein the antisense strand is 23 nucleotides in length and wherein the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 10, 11, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides and nucleotides 2, 4, 6, 8, 12, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 21 nucleotides in length and nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 11, 12, 13, 15, 17, 19, and 21 are 2′-fluoro nucleotides, nucleotides 2, 4, 6, 8, 10, 14, 16, 18, and 20 are 2′-O-methyl nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern of Pattern IX and the sense strand has the modification pattern of Pattern X.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and sense strand are not covalently linked (i.e. the antisense strand and sense strand form an siRNA), wherein the antisense strand is 23 nucleotides in length and wherein the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides and nucleotides 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 are 2′-fluoro nucleotides the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 23 nucleotides in length and wherein the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 21 are 2′-fluoro nucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 are 2′-O-methyl nucleotides, nucleotides 22 and 23 are beta-D-deoxynucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern represented by Pattern V and the sense strand has the modification represented by Pattern XI.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and sense strand are not covalently linked (i.e. the antisense strand and sense strand form an siRNA), wherein the antisense strand is 23 nucleotides in length and wherein the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 12, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides and nucleotides 2, 4, 6, 8, 10, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 23 nucleotides in length and wherein the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 10, 11, 13, 15, 17, 19, and 21 are 2′-fluoro nucleotides, nucleotides 2, 4, 6, 8, 12, 14, 16, 18, and 20 are 2′-O-methyl nucleotides, nucleotides 22 and 23 are beta-D-deoxynucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern represented by Pattern VII and the sense strand has the modification pattern represented by Pattern XII.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and the sense strand are not covalently linked (i.e. the antisense strand and sense strand form an siRNA), wherein the antisense strand is 23 nucleotides in length and wherein the nucleotides of the antisense strand are modified such, that counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 10, 11, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides and nucleotides 2, 4, 6, 8, 12, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 23 nucleotides in length and wherein the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 3, 5, 7, 9, 11, 12, 13, 15, 17, 19, and 21 are 2′-fluoro nucleotides, nucleotides 2, 4, 6, 8, 10, 14, 16, 18, and 20 are 2′-O-methyl nucleotides, nucleotides 22 and 23 are beta-D-deoxynucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern of Pattern IX and the sense strand has the modification pattern of Pattern XIII.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and the sense strand are not covalently linked (i.e., the antisense strand and sense strand from an siRNA), wherein the antisense strand is 23 nucleotides in length and wherein the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 21 nucleotides in length and wherein the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 2, 3, 4, 6, 8, 12, 14, 16, 18, 19, 20, and 21 are 2′-O-methyl nucleotides, nucleotides 5, 7, 9, 10, 11, 13, 15, and 17 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern of Pattern V and the sense strand has the modification pattern of Pattern XIV.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and the sense strand are not covalently linked (i.e., the antisense strand and sense strand from an siRNA), wherein the antisense strand is 23 nucleotides in length and the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 8, 9, 11, 12, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides, nucleotides 2, 4, 6, 10, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 2, 3, 4, 5, 6, 8, 12, 14, 16, 18, 19, 20, and 21 are 2′-O-methyl nucleotides, nucleotides 7, 9, 10, 11, 13, 15, and 17 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern of Pattern XVI and the sense strand has the modification pattern of Pattern XV.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and the sense strand are not covalently linked (i.e., the antisense strand and sense strand from an siRNA), wherein the antisense strand is 23 nucleotides in length and the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 8, 9, 11, 12, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides, nucleotides 2, 4, 6, 10, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21 are 2′-O-methyl nucleotides, nucleotides 7, 9, 10, and 11 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern of Pattern XVII and the sense strand has the modification pattern of Pattern XVIII.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and the sense strand are not covalently linked (i.e., the antisense strand and sense strand from an siRNA), wherein the antisense strand is 23 nucleotides in length and the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 8, 9, 11, 12, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides, nucleotides 2, 4, 6, 10, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 21 nucleotides in length and the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 2, 3, 4, 5, 6, 8, 12, 13, 14, 15, 16, 17, 18, 19, 20, and 21 are 2′-O-methyl nucleotides, nucleotides 7, 9, 10, and 11 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern of Pattern XVII and the sense strand has the modification pattern of Pattern XIX.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and the sense strand are not covalently linked (i.e., the antisense strand and sense strand from an siRNA), wherein the antisense strand is 23 nucleotides in length and wherein the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 21 nucleotides in length and wherein the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1 and 2 are 2′-O-methoxyethyl nucleotides, nucleotides 3, 4, 6, 8, 12, 14, 16, 18, 19, 20, and 21 are 2′-O-methyl nucleotides, nucleotides 5, 7, 9, 10, 11, 13, 15, and 17 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern of Pattern V and the sense strand has the modification pattern of Pattern XX.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and the sense strand are not covalently linked (i.e., the antisense strand and sense strand from an siRNA), wherein the antisense strand is 23 nucleotides in length and wherein the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 21 nucleotides in length and wherein the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 2 and 3 are 2′-O-methoxyethyl nucleotides, nucleotides 1, 4, 6, 8, 12, 14, 16, 18, 19, 20, and 21 are 2′-O-methyl nucleotides, nucleotides 5, 7, 9, 10, 11, 13, 15, and 17 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern of Pattern V and the sense strand has the modification pattern of Pattern XXI.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and the sense strand are not covalently linked (i.e., the antisense strand and sense strand from an siRNA), wherein the antisense strand is 23 nucleotides in length and wherein the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 21 nucleotides in length and wherein the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 2, 3, 19 and 20 are 2′-O-methoxyethyl nucleotides, nucleotides 1, 4, 6, 8, 12, 14, 16, 18, and 21 are 2′-O-methyl nucleotides, nucleotides 5, 7, 9, 10, 11, 13, 15, and 17 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern of Pattern V and the sense strand has the modification pattern of Pattern XXII.
In embodiments, a compound comprises an antisense strand and a sense strand hybridized to form a double-stranded nucleic acid, wherein the antisense strand and the sense strand are not covalently linked (i.e., the antisense strand and sense strand from an siRNA), wherein the antisense strand is 23 nucleotides in length and wherein the nucleotides of the antisense strand are modified such that, counting from the 5′ terminus of the antisense strand, nucleotides 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 22, and 23 are 2′-O-methyl nucleotides, nucleotides 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage; and wherein the sense strand is 21 nucleotides in length and wherein the nucleotides of the sense strand are modified such that, counting from the 5′ terminus of the sense strand, nucleotides 1, 2, 3, and 4 are 2′-O-methoxyethyl nucleotides, nucleotides 6, 8, 12, 14, 16, 18, 19, 20 and 21 are 2′-O-methyl nucleotides, nucleotides 5, 7, 9, 10, 11, 13, 15, and 17 are 2′-fluoro nucleotides, the first two internucleotide linkages at the 5′ terminus and the last two internucleotide linkages at the 3′ terminus are phosphorothioate internucleotide linkages, and each other internucleotide linkage is a phosphodiester internucleotide linkage. In such embodiments, the antisense strand has the modification pattern of Pattern V and the sense strand has the modification pattern of Pattern XXIV.
In embodiments, a compound provided herein comprises a covalently linked ligand.
In embodiments, a compound provided herein comprises a ligand covalently linked to the antisense strand. In embodiments, a compound provided herein comprises a ligand covalently linked to the sense strand. In embodiments, the ligand comprises an uptake motif with one or more long chain fatty acids (LFCA).
In embodiments, a compound comprising an uptake motif has the structure (I)
wherein A is a double-stranded nucleic acid and t is an integer from 1 to 5. In embodiments, A is the sense strand. In embodiments, A is the antisense strand.
L3 and L4 are independently a bond, —N(R23)—, —O—, —S—, —C(O)—, —N(R23)C(O)—, —C(O)N(R24)—, —N(R23)C(O)N(R24)—, —C(O)O—, —OC(O)—, —N(R23)C(O)O—, —OC(O)N(R24)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R25)—O—, —O—P(S)(R25)—O—, —O—P(O)(NR23R24)—N—, —O—P(S)(NR23R24)—N—, —O—P(O)(NR23R24)—O—, —O—P(S)(NR23R24)—O—, —P(O)(NR23R24)—N—, —P(S)(NR23R24)—N—, —P(O)(NR23R24)—O—, —P(S)(NR23R24)—O—, —S—S—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene or substituted or unsubstituted heteroarylene. Each R23, R24 and R25 is independently hydrogen or unsubstituted C1-C10 alkyl.
L5 is -L5A-L5B-L5C-L5D-L5E- and L6 is -L6A-L6B-L6C-L6D-L6E. L5A, L5B, L5C, L5D, L5E, L6A, L6B L6C, L6D, and L6E are independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene or substituted or unsubstituted heteroarylene.
R1 and R2 are independently unsubstituted C1-C25 alkyl, wherein at least one of R1 and R2 is unsubstituted C9-C19 alkyl. In embodiments, R1 and R2 are independently unsubstituted C1-C20 alkyl, wherein at least one of R1 and R2 is unsubstituted C9-C19 alkyl.
R3 is hydrogen, -hydrogen, —NH2, —OH, —SH, —C(O)H, —C(O)NH2, —NHC(O)H, —NHC(O)OH, —NHC(O)NH2, —C(O)OH, —OC(O)H, —N3, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
In embodiments, t is 1. In embodiments, t is 2. In embodiments, t is 3. In embodiments, t is 4. In embodiments, t is 5.
In embodiments, one L3 is attached to a 3′ carbon of a nucleotide. In embodiments, one L3 is attached to the 3′ carbon the 3′ terminal nucleotide of the sense strand. In embodiments, one L3 is attached to the 3′ carbon of the 3′ terminal nucleotide of the antisense strand.
In embodiments, one L3 is attached to a 5′ carbon of a nucleotide. In embodiments, one L3 is attached to the 5′ carbon of the 5′ terminal nucleotide of the sense strand. In embodiments, one L3 is attached to the 5′ carbon of the 5′ terminal nucleotide of the antisense strand.
In embodiments, one L3 is attached to a 2′ carbon of a nucleotide. In embodiments, one L3 is attached to a 2′ carbon of a nucleotide of the sense strand. In embodiments, one L3 is attached to a 2′ carbon of a nucleotide of the antisense strand.
In embodiments, one L3 is attached to a nucleobase. In embodiments, one L3 is attached to a nucleobase of the sense strand. In embodiments, one L3 is attached to a nucleobase of the antisense strand.
In embodiments, one L3 is attached to a phosphate group at a 3′ carbon of a nucleotide. In embodiments, one L3 is attached to a phosphate group at the 3′ carbon the 3′ terminal nucleotide of the sense strand. In embodiments, one L3 is attached to a phosphate group at the 3′ carbon of the 3′ terminal nucleotide of the antisense strand.
In embodiments, one L3 is attached to a phosphate group at a 5′ carbon of a nucleotide. In embodiments, one L3 is attached to a phosphate group at the 5′ carbon of the 5′ terminal nucleotide of the sense strand. In embodiments, one L3 is attached to a phosphate group at the 5′ carbon of the 5′ terminal nucleotide of the antisense strand.
In embodiments, one L3 is attached to a phosphate group at a 2′ carbon of a nucleotide. In embodiments, one L3 is attached to a phosphate group at a 2′ carbon of a nucleotide of the sense strand. In embodiments, one L3 is attached to a phosphate group a 2′ carbon of a nucleotide of the antisense strand.
In embodiments, L3 is a bond, —N(R23)—, —O—, —S—, —C(O)—, —N(R23)C(O)—, —C(O)N(R24)—, —N(R23)C(O)N(R24)—, —C(O)O—, —OC(O)—, —N(R23)C(O)O—, —OC(O)N(R24)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R25)—O—, —O—P(S)(R25)—O—, —O—P(O)(NR23R24)—N—, —O—P(S)(NR23R24)—N—, —O—P(O)(NR23R24)—O—, —O—P(S)(NR23R24)—O—, —P(O)(NR23R24)—N—, —P(S)(NR23R24)—N—, —P(O)(NR23R24)—O—, —P(S)(NR23R24)—O—, —S—S—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene.
In embodiments, L3 is a bond. In embodiments, L3 is —N(R23)—. In embodiments, L3 is —O— or —S—. In embodiments, L3 is —C(O)—. In embodiments, L3 is —N(R23)C(O)— or —C(O)N(R24)—. In embodiments, L3 is —N(R23)C(O)N(R24)—. In embodiments, L3 is —C(O)O— or —OC(O)—. In embodiments, L3 is —N(R23)C(O)O— or —OC(O)N(R24)—. In embodiments, L3 is —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R25)—O—, —O—P(O)(NR23R24)—N—, or —O—P(O)(NR23R24)—O—.
In embodiments, L3 is —P(O)(NR23R24)—N—, —P(S)(NR23R24)—N—, —P(O)(NR23R24)—O— or —P(S)(NR23R24)—O—. In embodiments, L3 is —S—S—.
In embodiments, L3 is independently substituted or unsubstituted alkylene (e.g., C1-C23, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L3 is independently substituted alkylene (e.g., C1-C23, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L3 is independently unsubstituted alkylene (e.g., C1-C23, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L3 is independently substituted or unsubstituted C1-C23 alkylene. In embodiments, L3 is independently substituted C1-C23 alkylene. In embodiments, L3 is independently unsubstituted C1-C23 alkylene. In embodiments, L3 is independently substituted or unsubstituted C1-C12 alkylene. In embodiments, L3 is independently substituted C1-C12 alkylene. In embodiments, L3 is independently unsubstituted C1-C12 alkylene. In embodiments, L3 is independently substituted or unsubstituted C1-C8 alkylene. In embodiments, L3 is independently substituted C1-C8 alkylene. In embodiments, L3 is independently unsubstituted C1-C8 alkylene. In embodiments, L3 is independently substituted or unsubstituted C1-C6 alkylene. In embodiments, L3 is independently substituted C1-C6 alkylene. In embodiments, L3 is independently unsubstituted C1-C6 alkylene. In embodiments, L3 is independently substituted or unsubstituted C1-C4 alkylene. In embodiments, L3 is independently substituted C1-C4 alkylene. In embodiments, L3 is independently unsubstituted C1-C4 alkylene. In embodiments, L3 is independently substituted or unsubstituted ethylene. In embodiments, L3 is independently substituted ethylene. In embodiments, L3 is independently unsubstituted ethylene. In embodiments, L3 is independently substituted or unsubstituted methylene. In embodiments, L3 is independently substituted methylene. In embodiments, L3 is independently unsubstituted methylene.
In embodiments, L3 is independently substituted or unsubstituted heteroalkylene (e.g., 2 to 23 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In embodiments, L3 is independently substituted heteroalkylene (e.g., 2 to 23 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In embodiments, L3 is independently unsubstituted heteroalkylene (e.g., 2 to 23 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In embodiments, L3 is independently substituted or unsubstituted 2 to 23 membered heteroalkylene. In embodiments, L3 is independently substituted 2 to 23 membered heteroalkylene. In embodiments, L3 is independently unsubstituted 2 to 23 membered heteroalkylene. In embodiments, L3 is independently substituted or unsubstituted 2 to 8 membered heteroalkylene. In embodiments, L3 is independently substituted 2 to 8 membered heteroalkylene. In embodiments, L3 is independently unsubstituted 2 to 8 membered heteroalkylene. In embodiments, L3 is independently substituted or unsubstituted 2 to 6 membered heteroalkylene. In embodiments, L3 is independently substituted 2 to 6 membered heteroalkylene. In embodiments, L3 is independently unsubstituted 2 to 6 membered heteroalkylene. In embodiments, L3 is independently substituted or unsubstituted 4 to 6 membered heteroalkylene. In embodiments, L3 is independently substituted 4 to 6 membered heteroalkylene. In embodiments, L3 is independently unsubstituted 4 to 6 membered heteroalkylene. In embodiments, L3 is independently substituted or unsubstituted 2 to 3 membered heteroalkylene. In embodiments, L3 is independently substituted 2 to 3 membered heteroalkylene. In embodiments, L3 is independently unsubstituted 2 to 3 membered heteroalkylene. In embodiments, L3 is independently substituted or unsubstituted 4 to 5 membered heteroalkylene. In embodiments, L3 is independently substituted 4 to 5 membered heteroalkylene. In embodiments, L3 is independently unsubstituted 4 to 5 membered heteroalkylene.
In embodiments, L4 is a bond, —N(R23)—, —O—, —S—, —C(O)—, —N(R23)C(O)—, —C(O)N(R24)—, —N(R23)C(O)N(R24)—, —C(O)O—, —OC(O)—, —N(R23)C(O)O—, —OC(O)N(R24)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R25)—O—, —O—P(S)(R25)—O—, —O—P(O)(NR23R24)—N—, —O—P(S)(NR23R24)—N—, —O—P(O)(NR23R24)—O—, —O—P(S)(NR23R24)—O—, —P(O)(NR23R24)—N—, —P(S)(NR23R24)—N—, —P(O)(NR23R24)—O—, —P(S)(NR23R24)—O—, —S—S—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene.
In embodiments, L4 is a bond. In embodiments, L4 is —N(R23)—. In embodiments, L4 is —O— or —S—. In embodiments, L4 is —C(O)—. In embodiments, L4 is —N(R23)C(O)— or —C(O)N(R24)—. In embodiments, L4 is —N(R23)C(O)N(R24)—. In embodiments, L4 is —C(O)O— or —OC(O)—. In embodiments, L4 is —N(R23)C(O)O— or —OC(O)N(R24)—. In embodiments, L4 is —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R25)—O—, —O—P(O)(NR23R24)—N—, or —O—P(O)(NR23R24)—O—. In embodiments, L4 is —P(O)(NR23R24)—N—, —P(S)(NR23R24)—N—, —P(O)(NR23R24)—O— or —P(S)(NR23R24)—O—. In embodiments, L4 is —S—S—.
In embodiments, L4 is independently substituted or unsubstituted alkylene (e.g., C1-C23, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L4 is independently substituted alkylene (e.g., C1-C23, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L4 is independently unsubstituted alkylene (e.g., C1-C23, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L4 is independently substituted or unsubstituted C1-C23 alkylene. In embodiments, L4 is independently substituted C1-C23 alkylene. In embodiments, L4 is independently unsubstituted C1-C23 alkylene. In embodiments, L4 is independently substituted or unsubstituted C1-C12 alkylene. In embodiments, L4 is independently substituted C1-C12 alkylene. In embodiments, L4 is independently unsubstituted C1-C12 alkylene. In embodiments, L4 is independently substituted or unsubstituted C1-C8 alkylene. In embodiments, L4 is independently substituted C1-C8 alkylene. In embodiments, L4 is independently unsubstituted C1-C8 alkylene. In embodiments, L4 is independently substituted or unsubstituted C1-C6 alkylene. In embodiments, L4 is independently substituted C1-C6 alkylene. In embodiments, L4 is independently unsubstituted C1-C6 alkylene. In embodiments, L4 is independently substituted or unsubstituted C1-C4 alkylene. In embodiments, L4 is independently substituted C1-C4 alkylene. In embodiments, L4 is independently unsubstituted C1-C4 alkylene. In embodiments, L4 is independently substituted or unsubstituted ethylene. In embodiments, L4 is independently substituted ethylene. In embodiments, L4 is independently unsubstituted ethylene. In embodiments, L4 is independently substituted or unsubstituted methylene. In embodiments, L4 is independently substituted methylene. In embodiments, L4 is independently unsubstituted methylene.
In embodiments, L4 is independently substituted or unsubstituted heteroalkylene (e.g., 2 to 23 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In embodiments, L4 is independently substituted heteroalkylene (e.g., 2 to 23 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In embodiments, L4 is independently unsubstituted heteroalkylene (e.g., 2 to 23 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In embodiments, L4 is independently substituted or unsubstituted 2 to 23 membered heteroalkylene. In embodiments, L4 is independently substituted 2 to 23 membered heteroalkylene. In embodiments, L4 is independently unsubstituted 2 to 23 membered heteroalkylene. In embodiments, L4 is independently substituted or unsubstituted 2 to 8 membered heteroalkylene. In embodiments, L4 is independently substituted 2 to 8 membered heteroalkylene. In embodiments, L4 is independently unsubstituted 2 to 8 membered heteroalkylene. In embodiments, L4 is independently substituted or unsubstituted 2 to 6 membered heteroalkylene. In embodiments, L4 is independently substituted 2 to 6 membered heteroalkylene. In embodiments, L4 is independently unsubstituted 2 to 6 membered heteroalkylene. In embodiments, L4 is independently substituted or unsubstituted 4 to 6 membered heteroalkylene. In embodiments, L4 is independently substituted 4 to 6 membered heteroalkylene. In embodiments, L4 is independently unsubstituted 4 to 6 membered heteroalkylene. In embodiments, L4 is independently substituted or unsubstituted 2 to 3 membered heteroalkylene. In embodiments, L4 is independently substituted 2 to 3 membered heteroalkylene. In embodiments, L4 is independently unsubstituted 2 to 3 membered heteroalkylene. In embodiments, L4 is independently substituted or unsubstituted 4 to 5 membered heteroalkylene. In embodiments, L4 is independently substituted 4 to 5 membered heteroalkylene. In embodiments, L4 is independently unsubstituted 4 to 5 membered heteroalkylene.
R23 is independently hydrogen or unsubstituted alkyl (e.g., C1-C23, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R23 is independently hydrogen. In embodiments, R23 is independently unsubstituted C1-C23 alkyl. In embodiments, R23 is independently hydrogen or unsubstituted C1-C12 alkyl. In embodiments, R23 is independently hydrogen or unsubstituted C1-C10 alkyl. In embodiments, R23 is independently hydrogen or unsubstituted C1-C8 alkyl. In embodiments, R23 is independently hydrogen or unsubstituted C1-C6 alkyl. In embodiments, R23 is independently hydrogen or unsubstituted C1-C4 alkyl. In embodiments, R23 is independently hydrogen or unsubstituted C1-C2 alkyl.
R24 is independently hydrogen or unsubstituted alkyl (e.g., C1-C24, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R24 is independently hydrogen. In embodiments, R24 is independently unsubstituted C1-C24 alkyl. In embodiments, R24 is independently hydrogen or unsubstituted C1-C12 alkyl. In embodiments, R24 is independently hydrogen or unsubstituted C1-C10 alkyl. In embodiments, R24 is independently hydrogen or unsubstituted C1-C8 alkyl. In embodiments, R24 is independently hydrogen or unsubstituted C1-C6 alkyl. In embodiments, R24 is independently hydrogen or unsubstituted C1-C4 alkyl. In embodiments, R24 is independently hydrogen or unsubstituted C1-C2 alkyl.
R25 is independently hydrogen or unsubstituted alkyl (e.g., C1-C25, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R25 is independently hydrogen. In embodiments, R25 is independently unsubstituted C1-C25 alkyl. In embodiments, R25 is independently hydrogen or unsubstituted C1-C12 alkyl. In embodiments, R25 is independently hydrogen or unsubstituted C1-C10 alkyl. In embodiments, R25 is independently hydrogen or unsubstituted C1-C8 alkyl. In embodiments, R25 is independently hydrogen or unsubstituted C1-C6 alkyl. In embodiments, R25 is independently hydrogen or unsubstituted C1-C4 alkyl. In embodiments, R25 is independently hydrogen or unsubstituted C1-C2 alkyl.
In embodiments, L3 and L4 are independently a bond, —NH—, —O—, —C(O)—, —C(O)O—, —OC(O)—, —OPO2—O— —O—P(O)(S)—O—, —O—P(O)(CH3)—O—, —O—P(S)(CH3)—O—, —O—P(O)(N(CH3)2)—N—, —O—P(O)(N(CH3)2)—O—, —O—P(S)(N(CH3)2)—N—, —O—P(S)(N(CH3)2)—O—, —P(O)(N(CH3)2)—N—, —P(O)(N(CH3)2)—O—, —P(S)(N(CH3)2)—N—, —P(S)(N(CH3)2)—O—, substituted or unsubstituted alkylene or substituted or unsubstituted heteroalkylene. In embodiments, L3 is independently a bond, —NH—, —O—, —C(O)—, —C(O)O—, —OC(O)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(CH3)—O—, —O—P(S)(CH3)—O—, —O—P(O)(N(CH3)2)—N—, —O—P(O)(N(CH3)2)—O—, —O—P(S)(N(CH3)2)—N—, —O—P(S)(N(CH3)2)—O—, —P(O)(N(CH3)2)—N—, —P(O)(N(CH3)2)—O—, —P(S)(N(CH3)2)—N—, —P(S)(N(CH3)2)—O—, substituted or unsubstituted alkylene or substituted or unsubstituted heteroalkylene. In embodiments, L4 is independently a bond, —NH—, —O—, —C(O)—, —C(O)O—, —OC(O)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(CH3)—O—, —O—P(S)(CH3)—O—, —O—P(O)(N(CH3)2)—N—, —O—P(O)(N(CH3)2)—O—, —O—P(S)(N(CH3)2)—N—, —O—P(S)(N(CH3)2)—O—, —P(O)(N(CH3)2)—N—, —P(O)(N(CH3)2)—O—, —P(S)(N(CH3)2)—N—, —P(S)(N(CH3)2)—O—, substituted or unsubstituted alkylene or substituted or unsubstituted heteroalkylene.
In embodiments, L3 is independently
In embodiments, L3 is independently —OPO2—O—. In embodiments, L3 is independently —O—P(O)(S)—O—. In embodiments, L3 is independently —O—. In embodiments, L3 is independently —S—.
In embodiments, L4 is independently substituted or unsubstituted alkylene or substituted or unsubstituted heteroalkylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—. In embodiments, L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L7 is independently substituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L7 is independently unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2).
In embodiments, L4 is independently substituted or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L4 is independently substituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L4 is independently oxo-substituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L4 is independently unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered).
In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L4 is independently -L7-NH—C(O)—; and L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L4 is independently -L7-C(O)—NH—; and L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2).
In embodiments, L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L7 is independently substituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L7 is independently unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2).
In embodiments, L7 is independently substituted or unsubstituted C1-C20 alkylene. In embodiments, L7 is independently substituted C1-C20 alkylene. In embodiments, L7 is independently hydroxy(OH)-substituted C1-C20 alkylene. In embodiments, L7 is independently hydroxymethyl-substituted C1-C20 alkylene. In embodiments, L7 is independently unsubstituted C1-C20 alkylene. In embodiments, L7 is independently substituted or unsubstituted C1-C12 alkylene. In embodiments, L7 is independently substituted C1-C12 alkylene. In embodiments, L7 is independently hydroxy(OH)-substituted C1-C12 alkylene. In embodiments, L7 is independently hydroxymethyl-substituted C1-C12 alkylene. In embodiments, L7 is independently unsubstituted C1-C12 alkylene. In embodiments, L7 is independently substituted or unsubstituted C1-C8 alkylene. In embodiments, L7 is independently substituted C1-C8 alkylene. In embodiments, L7 is independently hydroxy(OH)-substituted C1-C8 alkylene. In embodiments, L7 is independently hydroxymethyl-substituted C1-C8 alkylene. In embodiments, L7 is independently unsubstituted C1-C8 alkylene. In embodiments, L7 is independently substituted or unsubstituted C1-C6 alkylene. In embodiments, L7 is independently substituted C1-C6 alkylene. In embodiments, L7 is independently hydroxy(OH)-substituted C1-C6 alkylene. In embodiments, L7 is independently hydroxymethyl-substituted C1-C6 alkylene. In embodiments, L7 is independently unsubstituted C1-C6 alkylene. In embodiments, L7 is independently substituted or unsubstituted C1-C4 alkylene. In embodiments, L7 is independently substituted C1-C4 alkylene. In embodiments, L7 is independently hydroxy(OH)-substituted C1-C4 alkylene. In embodiments, L7 is independently hydroxymethyl-substituted C1-C4 alkylene. In embodiments, L7 is independently unsubstituted C1-C4 alkylene. In embodiments, L7 is independently substituted or unsubstituted C1-C2 alkylene. In embodiments, L7 is independently substituted C1-C2 alkylene. In embodiments, L7 is independently hydroxy(OH)-substituted C1-C2 alkylene. In embodiments, L7 is independently hydroxymethyl-substituted C1-C2 alkylene. In embodiments, L7 is independently unsubstituted C1-C2 alkylene.
In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted or unsubstituted C1-C8 alkylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted C1-C8 alkylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted C1-C8 alkylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently hydroxymethyl-substituted C1-C8 alkylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently unsubstituted C1-C8 alkylene.
In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted or unsubstituted C3-C8 alkylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted C3-C8 alkylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted C3-C8 alkylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently hydroxymethyl-substituted C3-C8 alkylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently unsubstituted C3-C8 alkylene.
In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted or unsubstituted C5-C8 alkylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted C5-C8 alkylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted C5-C8 alkylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently hydroxymethyl-substituted C5-C8 alkylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently unsubstituted C5-C8 alkylene.
In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted or unsubstituted octylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted octylene.
In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted octylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently unsubstituted octylene. In embodiments, L4 is independently -L7-NH—C(O)— and L7 is independently hydroxy(OH)-substituted octylene. In embodiments, L4 is independently -L7-NH—C(O)— and L7 is independently hydroxymethyl-substituted octylene. In embodiments, L4 is independently -L7-NH—C(O)— and L7 is independently unsubstituted octylene.
In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted or unsubstituted heptylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted heptylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted heptylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently unsubstituted heptylene. In embodiments, L4 is independently -L7-NH—C(O)— and L7 is independently hydroxy(OH)-substituted heptylene. In embodiments, L4 is independently -L7-NH—C(O)— and L7 is independently hydroxymethyl-substituted heptylene. In embodiments, L4 is independently -L7-NH—C(O)— and L7 is independently unsubstituted heptylene.
In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted or unsubstituted hexylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted hexylene.
In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted hexylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently unsubstituted hexylene. In embodiments, L4 is independently -L7-NH—C(O)— and L7 is independently hydroxy(OH)-substituted hexylene. In embodiments, L4 is independently -L7-NH—C(O)— and L7 is independently hydroxymethyl-substituted hexylene. In embodiments, L4 is independently -L7-NH—C(O)— and L7 is independently unsubstituted hexylene.
In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted or unsubstituted pentylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently substituted pentylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted pentylene. In embodiments, L4 is independently -L7-NH—C(O)— or -L7-C(O)—NH—; and L7 is independently unsubstituted pentylene. In embodiments, L4 is independently -L7-NH—C(O)— and L7 is independently hydroxy(OH)-substituted pentylene. In embodiments, L4 is independently -L7-NH—C(O)— and L7 is independently hydroxymethyl-substituted pentylene. In embodiments, L4 is independently -L7-NH—C(O)— and L7 is independently unsubstituted pentylene.
In embodiments, L4 is independently
In embodiments, L4 is independently
In embodiments, L4 is independently
In embodiments, L4 is independently
In embodiments, L4 is independently
In embodiments, L4 is independently
In embodiments, L4 is independently
In embodiments, L4 is
In embodiments, L4 is independently
In embodiments, L4 is independently
In embodiments, L4 is independently
In embodiments, L4 is independently
In embodiments, -L3-L4- is independently -L7-NH—C(O)— or -L7-C(O)—NH—. In embodiments, L7 is independently substituted or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L7 is independently substituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L7 is independently oxo-substituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L7 is independently unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L7 is independently substituted or unsubstituted heteroalkenylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L7 is independently substituted heteroalkenylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L7 is independently oxo-substituted heteroalkenylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L7 is independently unsubstituted heteroalkenylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered).
In embodiments, L7 is independently substituted or unsubstituted 2 to 20 membered heteroalkylene. In embodiments, L7 is independently substituted 2 to 20 membered heteroalkylene. In embodiments, L7 is independently oxo-substituted 2 to 20 membered heteroalkylene. In embodiments, L7 is independently unsubstituted 2 to 20 membered heteroalkylene. In embodiments, L7 is independently substituted or unsubstituted 2 to 12 membered heteroalkylene. In embodiments, L7 is independently substituted 2 to 12 membered heteroalkylene. In embodiments, L7 is independently oxo-substituted 2 to 12 membered heteroalkylene. In embodiments, L7 is independently unsubstituted 2 to 12 membered heteroalkylene. In embodiments, L7 is independently substituted or unsubstituted 2 to 10 membered heteroalkylene. In embodiments, L7 is independently substituted 2 to 10 membered heteroalkylene. In embodiments, L7 is independently oxo-substituted 2 to 10 membered heteroalkylene. In embodiments, L7 is independently unsubstituted 2 to 10 membered heteroalkylene. In embodiments, L7 is independently substituted or unsubstituted 2 to 8 membered heteroalkylene. In embodiments, L7 is independently substituted 2 to 8 membered heteroalkylene. In embodiments, L7 is independently oxo-substituted 2 to 8 membered heteroalkylene. In embodiments, L7 is independently unsubstituted 2 to 8 membered heteroalkylene. In embodiments, L7 is independently substituted or unsubstituted 2 to 6 membered heteroalkylene. In embodiments, L7 is independently substituted 2 to 6 membered heteroalkylene. In embodiments, L7 is independently oxo-substituted 2 to 6 membered heteroalkylene. In embodiments, L7 is independently unsubstituted 2 to 6 membered heteroalkylene. In embodiments, L7 is independently substituted or unsubstituted 2 to 4 membered heteroalkylene. In embodiments, L7 is independently substituted 2 to 4 membered heteroalkylene. In embodiments, L7 is independently oxo-substituted 2 to 4 membered heteroalkylene. In embodiments, L7 is independently unsubstituted 2 to 4 membered heteroalkylene.
In embodiments, L7 is independently substituted or unsubstituted 2 to 20 membered heteroalkenylene. In embodiments, L7 is independently substituted 2 to 20 membered heteroalkenylene. In embodiments, L7 is independently oxo-substituted 2 to 20 membered heteroalkenylene. In embodiments, L7 is independently unsubstituted 2 to 20 membered heteroalkenylene. In embodiments, L7 is independently substituted or unsubstituted 2 to 12 membered heteroalkenylene. In embodiments, L7 is independently substituted 2 to 12 membered heteroalkenylene. In embodiments, L7 is independently oxo-substituted 2 to 12 membered heteroalkenylene. In embodiments, L7 is independently unsubstituted 2 to 12 membered heteroalkenylene. In embodiments, L7 is independently substituted or unsubstituted 2 to 10 membered heteroalkenylene. In embodiments, L7 is independently substituted 2 to 10 membered heteroalkenylene. In embodiments, L7 is independently oxo-substituted 2 to 10 membered heteroalkenylene. In embodiments, L7 is independently unsubstituted 2 to 10 membered heteroalkenylene. In embodiments, L7 is independently substituted or unsubstituted 2 to 8 membered heteroalkenylene. In embodiments, L7 is independently substituted 2 to 8 membered heteroalkenylene. In embodiments, L7 is independently oxo-substituted 2 to 8 membered heteroalkenylene. In embodiments, L7 is independently unsubstituted 2 to 8 membered heteroalkenylene. In embodiments, L7 is independently substituted or unsubstituted 2 to 6 membered heteroalkenylene. In embodiments, L7 is independently substituted 2 to 6 membered heteroalkenylene. In embodiments, L7 is independently oxo-substituted 2 to 6 membered heteroalkenylene. In embodiments, L7 is independently unsubstituted 2 to 6 membered heteroalkenylene. In embodiments, L7 is independently substituted or unsubstituted 2 to 4 membered heteroalkenylene. In embodiments, L7 is independently substituted 2 to 4 membered heteroalkenylene. In embodiments, L7 is independently oxo-substituted 2 to 4 membered heteroalkenylene. In embodiments, L7 is independently unsubstituted 2 to 4 membered heteroalkenylene.
In embodiments, -L3-L4- is independently —O-L7-NH—C(O)— or —O-L7-C(O)—NH—. In embodiments, L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, -L3-L4- is independently —O-L7-NH—C(O)— or —O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, -L3-L4- is independently —O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2).
In embodiments, L3-L4- is independently —O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-C(O)—NH—; and L7 is independently substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-C(O)—NH— and L7 is independently hydroxymethyl-substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-C(O)—NH—; and L7 is independently unsubstituted C1-C8 alkylene.
In embodiments, -L3-L4- is independently —O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted C3-C8 alkylene. In embodiments, -L3-L4- is independently O-L7-C(O)—NH—; and L7 is independently substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-C(O)—NH— and L7 is independently hydroxymethyl-substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-C(O)—NH—; and L7 is independently unsubstituted C3-C8 alkylene.
In embodiments, -L3-L4- is independently —O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-C(O)—NH—; and L7 is independently substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-C(O)—NH— and L7 is independently hydroxymethyl-substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-C(O)—NH—; and L7 is independently unsubstituted C5-C8 alkylene.
In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently hydroxy(OH)-substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently hydroxymethyl-substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently unsubstituted C1-C8 alkylene.
In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently hydroxy(OH)-substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently hydroxymethyl-substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently unsubstituted C3-C8 alkylene.
In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently hydroxy(OH)-substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently hydroxymethyl-substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —O-L7-NH—C(O)—; and L7 is independently unsubstituted C5-C8 alkylene.
In embodiments, -L3-L4- is independently
In embodiments, -L3-L4- is independently
In embodiments, -L3-L4- is independently
In embodiments, -L3-L4- is independently
In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—, —OP(O)(S)—O-L7-NH—C(O)—, —OPO2—O-L7-C(O)—NH— or —OP(O)(S)—O-L7-C(O)—NH—. In embodiments, L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)— or —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH— or —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted alkylene.
In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)— or —OPO2—O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2).
In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)— or —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2).
In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently hydroxymethyl-substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently unsubstituted C1-C8 alkylene.
In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently hydroxymethyl-substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently unsubstituted C1-C8 alkylene.
In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently hydroxymethyl-substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently unsubstituted C3-C8 alkylene.
In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently hydroxymethyl-substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently unsubstituted C3-C8 alkylene.
In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently hydroxymethyl-substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-C(O)—NH—; and L7 is independently unsubstituted C5-C8 alkylene.
In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently substituted or unsubstituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently hydroxy(OH)-substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently hydroxymethyl-substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-C(O)—NH—; and L7 is independently unsubstituted C5-C8 alkylene.
In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently hydroxy(OH)-substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently hydroxymethyl-substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently unsubstituted C1-C8 alkylene.
In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)2—O-L7-NH—C(O)—; and L7 is independently hydroxy(OH)-substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently hydroxymethyl-substituted C1-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently unsubstituted C1-C8 alkylene.
In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently hydroxy(OH)-substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently hydroxymethyl-substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently unsubstituted C3-C8 alkylene.
In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently hydroxy(OH)-substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently hydroxymethyl-substituted C3-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently unsubstituted C3-C8 alkylene.
In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently hydroxy(OH)-substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently hydroxymethyl-substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OPO2—O-L7-NH—C(O)—; and L7 is independently unsubstituted C5-C8 alkylene.
In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently substituted or unsubstituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently hydroxy(OH)-substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently hydroxymethyl-substituted C5-C8 alkylene. In embodiments, -L3-L4- is independently —OP(O)(S)—O-L7-NH—C(O)—; and L7 is independently unsubstituted C5-C8 alkylene.
In embodiments, -L3-L4- is attached to a 3′ carbon of a nucleotide of the sense strand. In embodiments, -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand. In embodiments, -L3-L4- is attached to a 3′ carbon of the antisense sense strand. In embodiments, -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the antisense sense strand.
In embodiments, -L3-L4- is attached to a 5′ carbon of a nucleotide of the sense strand. In embodiments, -L3-L4- is attached to the 5′ carbon of the 5′ terminal nucleotide of the sense strand. In embodiments, -L3-L4- is attached to a 5′ carbon of a nucleotide of the antisense strand. In embodiments, -L3-L4- is attached to the 5′ carbon of the 5′ terminal nucleotide of the antisense strand.
In embodiments, -L3-L4- is attached to a 2′ carbon of a nucleotide of the sense strand. In embodiments, -L3-L4- is attached to a 2′ carbon of a nucleotide of the antisense strand.
In embodiments, -L3-L4- is attached to a nucleobase of the sense strand. In embodiments, -L3-L4- is attached to a nucleobase of the antisense strand.
In embodiments, -L3-L4- is independently
In embodiments, -L3-L4- is independently
In embodiments, -L3-L4- is independently
In embodiments, -L3-L4- is independently
In embodiments, -L3-L4- is independently
In embodiments, -L3-L4- is independently
In embodiments, -L3-L4- is independently
and is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand.
In embodiments, -L3-L4- is independently
and is attached to the 3′ carbon of the 3′ terminal nucleotide of the antisense strand.
In embodiments, -L3-L4- is independently
that is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand.
In embodiments, -L3-L4- is independently
that is attached to the 3′ carbon of the 3′ terminal nucleotide of the antisense strand.
In embodiments, -L3-L4- is independently
that is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand.
In embodiments, -L3-L4- is independently
that is attached to the 3′ carbon of the 3′ terminal nucleotide of the antisense strand.
In embodiments, an -L3-L4- is independently
and is attached to the 5′ carbon of the 5′ terminal nucleotide of the sense strand.
In embodiments, an -L3-L4- is independently 0
and is attached to the 5′ carbon of the 5′ terminal nucleotide of the antisense strand.
In embodiments, an -L3-L4- is independently
that is attached to the 5′ carbon of the 5′ terminal nucleotide of the sense strand.
In embodiments, an -L3-L4- is independently
that is attached to the 5′ carbon of the 5′ terminal nucleotide of the antisense strand.
In embodiments, an -L3-L4- is independently
that is attached to 5′ carbon of the 5′ terminal nucleotide of the sense strand.
In embodiments, an -L3-L4- is independently
that is attached to the 5′ carbon of the 5′ terminal nucleotide of the antisense strand.
In embodiments, an -L3-L4- is independently attached to a nucleobase of the sense strand. In embodiments, an -L3-L4- is independently
and is attached to a nucleobase of the sense strand.
In embodiments, an -L3-L4- is independently
and is attached to a nucleobase of the antisense strand.
In embodiments, -L3-L4- is independently
In embodiments, -L3-L4- is independently
that is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand.
In embodiments, -L3-L4- is independently
that is attached to the 3′ carbon of the 3′ terminal nucleotide of the antisense strand.
In embodiments, -L3-L4- is independently
that is attached to the 5′ carbon of the 5′ terminal nucleotide of the sense strand.
In embodiments, -L3-L4- is independently
that is attached to the 5′ carbon of the 5′ terminal nucleotide of the antisense strand.
In embodiments, -L3-L4- is independently
that is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand.
In embodiments, -L3-L4- is independently
that is attached to the 3′ carbon of the 3′ terminal nucleotide of the antisense strand.
In embodiments, -L3-L4- is independently
that is attached to the 5′ carbon of the 5′ terminal nucleotide of the sense strand.
In embodiments, -L3-L4- is independently
that is attached to the 5′ carbon of the 5′ terminal nucleotide of the antisense strand.
In embodiments, -L3-L4- is independently
that is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand.
In embodiments, -L3-L4- is independently
that is attached to the 3′ carbon of the 3′ terminal nucleotide of the antisense strand.
In embodiments, -L3-L4- is independently
and is attached to the 5′ carbon of the 5′ terminal nucleotide of the sense strand.
In embodiments, -L3-L4- is independently
and is attached to the 5′ carbon of the 5′ terminal nucleotide of the antisense strand.
In embodiments, -L3-L4- is independently
and is attached to a 2′ carbon of a nucleotide of the sense strand.
In embodiments, -L3-L4- is independently
and is attached to a 2′ carbon of a nucleotide of the antisense strand.
In embodiments, -L3-L4- is independently
and is attached to a 2′ carbon of a nucleotide of the sense strand.
In embodiments, -L3-L4- is independently
and is attached to a 2′ carbon of a nucleotide of the antisense strand.
In embodiments, -L3-L4- is independently
and is attached to a nucleobase of the sense strand.
In embodiments, -L3-L4- is independently
and is attached to a nucleobase of the antisense strand.
In embodiments, R3 is independently hydrogen, —NH2, —OH, —SH, —C(O)H, —C(O)NH2, —NHC(O)H, —NHC(O)OH, —NHC(O)NH2, —C(O)OH, —OC(O)H, —N3, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl. In embodiments, R3 is independently hydrogen. In embodiments, R3 is independently —NH2. In embodiments, R3 is independently —OH. In embodiments, R3 is independently —SH. In embodiments, R3 is independently —C(O)H. In embodiments, R3 is independently —C(O)NH2. In embodiments, R3 is independently —NHC(O)H. In embodiments, R3 is independently —NHC(O)OH. In embodiments, R3 is independently —NHC(O)NH2. In embodiments, R3 is independently —C(O)OH. In embodiments, R3 is independently —OC(O)H. In embodiments, R3 is independently —N3.
In embodiments, R3 is independently substituted or unsubstituted alkyl (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R3 is independently substituted or unsubstituted C1-C20 alkyl. In embodiments, R3 is independently substituted C1-C20 alkyl. In embodiments, R3 is independently unsubstituted C1-C20 alkyl. In embodiments, R3 is independently substituted or unsubstituted C1-C12 alkyl. In embodiments, R3 is independently substituted C1-C12 alkyl. In embodiments, R3 is independently unsubstituted C1-C12 alkyl. In embodiments, R3 is independently substituted or unsubstituted C1-C8 alkyl. In embodiments, R3 is independently substituted C1-C8 alkyl. In embodiments, R3 is independently unsubstituted C1-C8 alkyl. In embodiments, R3 is independently substituted or unsubstituted C1-C6 alkyl. In embodiments, R3 is independently substituted C1-C6 alkyl. In embodiments, R3 is independently unsubstituted C1-C6 alkyl. In embodiments, R3 is independently substituted or unsubstituted C1-C4 alkyl. In embodiments, R3 is independently substituted C1-C4 alkyl. In embodiments, R3 is independently unsubstituted C1-C4 alkyl. In embodiments, R3 is independently substituted or unsubstituted ethyl. In embodiments, R3 is independently substituted ethyl. In embodiments, R3 is independently unsubstituted ethyl. In embodiments, R3 is independently substituted or unsubstituted methyl. In embodiments, R3 is independently substituted methyl. In embodiments, R3 is independently unsubstituted methyl.
In embodiments, L6 is independently —NHC(O)—. In embodiments, L6 is independently —C(O)NH—. In embodiments, L6 is independently substituted or unsubstituted alkylene. In embodiments, L6 is independently substituted or unsubstituted heteroalkylene.
In embodiments, L6 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L6 is independently substituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L6 is independently unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2).
In embodiments, L6 is independently substituted or unsubstituted C1-C20 alkylene. In embodiments, L6 is independently substituted C1-C20 alkylene. In embodiments, L6 is independently unsubstituted C1-C20 alkylene. In embodiments, L6 is independently substituted or unsubstituted C1-C12 alkylene. In embodiments, L6 is independently substituted C1-C12 alkylene. In embodiments, L6 is independently unsubstituted C1-C12 alkylene. In embodiments, L6 is independently substituted or unsubstituted C1-C8 alkylene. In embodiments, L6 is independently substituted C1-C8 alkylene. In embodiments, L6 is independently unsubstituted C1-C8 alkylene. In embodiments, L6 is independently substituted or unsubstituted C1-C6 alkylene. In embodiments, L6 is independently substituted C1-C6 alkylene. In embodiments, L6 is independently unsubstituted C1-C6 alkylene. In embodiments, L6 is independently substituted or unsubstituted C1-C4 alkylene. In embodiments, L6 is independently substituted C1-C4 alkylene. In embodiments, L6 is independently unsubstituted C1-C4 alkylene. In embodiments, L6 is independently substituted or unsubstituted ethylene. In embodiments, L6 is independently substituted ethylene. In embodiments, L6 is independently unsubstituted ethylene. In embodiments, L6 is independently substituted or unsubstituted methylene. In embodiments, L6 is independently substituted methylene. In embodiments, L6 is independently unsubstituted methylene.
In embodiments, L6 is independently substituted or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In embodiments, L6 is independently substituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In embodiments, L6 is independently unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In embodiments, L6 is independently substituted or unsubstituted 2 to 20 membered heteroalkylene. In embodiments, L6 is independently substituted 2 to 20 membered heteroalkylene. In embodiments, L6 is independently unsubstituted 2 to 20 membered heteroalkylene. In embodiments, L6 is independently substituted or unsubstituted 2 to 8 membered heteroalkylene. In embodiments, L6 is independently substituted 2 to 8 membered heteroalkylene. In embodiments, L6 is independently unsubstituted 2 to 8 membered heteroalkylene. In embodiments, L6 is independently substituted or unsubstituted 2 to 6 membered heteroalkylene. In embodiments, L6 is independently substituted 2 to 6 membered heteroalkylene. In embodiments, L6 is independently unsubstituted 2 to 6 membered heteroalkylene. In embodiments, L6 is independently substituted or unsubstituted 4 to 6 membered heteroalkylene. In embodiments, L6 is independently substituted 4 to 6 membered heteroalkylene. In embodiments, L6 is independently unsubstituted 4 to 6 membered heteroalkylene. In embodiments, L6 is independently substituted or unsubstituted 2 to 3 membered heteroalkylene. In embodiments, L6 is independently substituted 2 to 3 membered heteroalkylene. In embodiments, L6 is independently unsubstituted 2 to 3 membered heteroalkylene. In embodiments, L6 is independently substituted or unsubstituted 4 to 5 membered heteroalkylene. In embodiments, L6 is independently substituted 4 to 5 membered heteroalkylene. In embodiments, L6 is independently unsubstituted 4 to 5 membered heteroalkylene.
In embodiments, L6A is independently a bond or unsubstituted alkylene; L6B is independently a bond, —NHC(O)—, or unsubstituted arylene; L6C is independently a bond, unsubstituted alkylene, or unsubstituted arylene; L6D is independently a bond or unsubstituted alkylene; and L6E is independently a bond or —NHC(O)—. In embodiments, L6A is independently a bond or unsubstituted alkylene. In embodiments, L6B is independently a bond, —NHC(O)—, or unsubstituted arylene. In embodiments, L6C is independently a bond, unsubstituted alkylene, or unsubstituted arylene. In embodiments, L6D is independently a bond or unsubstituted alkylene. In embodiments, L6E is independently a bond or —NHC(O)—.
In embodiments, L6A is independently a bond or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L6A is independently unsubstituted C1-C20 alkylene. In embodiments, L6A is independently unsubstituted C1-C12 alkylene. In embodiments, L6A is independently unsubstituted C1-C8 alkylene. In embodiments, L6A is independently unsubstituted C1-C6 alkylene. In embodiments, L6A is independently unsubstituted C1-C4 alkylene. In embodiments, L6A is independently unsubstituted ethylene.
In embodiments, L6A is independently unsubstituted methylene. In embodiments, L6A is independently a bond.
In embodiments, L6B is independently a bond. In embodiments, L6B is independently —NHC(O)—. In embodiments, L6B is independently unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl). In embodiments, L6B is independently unsubstituted C6-C12 arylene. In embodiments, L6B is independently unsubstituted C6-C10 arylene. In embodiments, L6B is independently unsubstituted phenylene. In embodiments, L6B is independently unsubstituted naphthylene. In embodiments, L6B is independently unsubstituted biphenylene.
In embodiments, L6C is independently a bond or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L6C is independently unsubstituted C1-C20 alkylene. In embodiments, L6C is independently unsubstituted C1-C12 alkylene. In embodiments, L6C is independently unsubstituted C1-C8 alkylene. L6C is independently unsubstituted C2-C8 alkynylene. In embodiments, L6C is independently unsubstituted C1-C6 alkylene. In embodiments, L6C is independently unsubstituted C1-C4 alkylene. In embodiments, L6C is independently unsubstituted ethylene. In embodiments, L6C is independently unsubstituted methylene. In embodiments, L6C is independently a bond or unsubstituted alkynylene (e.g., C2-C20, C2-C12, C2-C8, C2-C6, C2-C4, or C2-C2). In embodiments, L6C is independently unsubstituted C2-C20 alkynylene. In embodiments, L6C is independently unsubstituted C2-C12 alkynylene. In embodiments, L6C is independently unsubstituted C2-C8 alkynylene. In embodiments, L6C is independently unsubstituted C2-C6 alkynylene. In embodiments, L6C is independently unsubstituted C2-C4 alkynylene. In embodiments, L6C is independently unsubstituted ethynylene. In embodiments, L6C is independently unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl). In embodiments, L6C is independently unsubstituted C6-C12 arylene. In embodiments, L6C is independently unsubstituted C6-C10 arylene. In embodiments, L6C is independently unsubstituted phenylene. In embodiments, L6C is independently unsubstituted naphthylene. In embodiments, L6C is independently a bond.
In embodiments, L6D is independently a bond or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L6D is independently unsubstituted C1-C20 alkylene. In embodiments, L6D is independently unsubstituted C1-C12 alkylene. In embodiments, L6A is independently unsubstituted C1-C8 alkylene. In embodiments, L6D is independently unsubstituted C1-C6 alkylene. In embodiments, L6D is independently unsubstituted C1-C4 alkylene. In embodiments, L6D is independently unsubstituted ethylene. In embodiments, L6D is independently unsubstituted methylene. In embodiments, L6D is independently a bond.
In embodiments, L6E is independently a bond. In embodiments, L6E is independently —NHC(O)—.
In embodiments, L6A is independently a bond or unsubstituted C1-C8 alkylene. In embodiments, L6B is independently a bond, —NHC(O)—, or unsubstituted phenylene. In embodiments, L6C is independently a bond, unsubstituted C2-C8 alkynylene, or unsubstituted phenylene. In embodiments, L6D is independently a bond or unsubstituted C1-C8 alkylene. In embodiments, L6E is independently a bond or —NHC(O)—.
In embodiments, L6 is independently a bond,
In embodiments, L6 is
independently a bond. In embodiments, L6 is independently
In embodiments, L6 is independently
In embodiments, L6 is independently
In embodiments, L6 is independently
In embodiments, L6 is independently
In embodiments, L5 is independently —NHC(O)—. In embodiments, L5 is independently —C(O)NH—. In embodiments, L5 is independently substituted or unsubstituted alkylene. In embodiments, L5 is independently substituted or unsubstituted heteroalkylene.
In embodiments, L5 is independently substituted or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L5 is independently substituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L5 is independently unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L5 is independently substituted or unsubstituted C1-C20 alkylene. In embodiments, L5 is independently substituted C1-C20 alkylene. In embodiments, L5 is independently unsubstituted C1-C20 alkylene. In embodiments, L5 is independently substituted or unsubstituted C1-C12 alkylene. In embodiments, L5 is independently substituted C1-C12 alkylene. In embodiments, L5 is independently unsubstituted C1-C12 alkylene. In embodiments, L5 is independently substituted or unsubstituted C1-C8 alkylene. In embodiments, L5 is independently substituted C1-C8 alkylene. In embodiments, L5 is independently unsubstituted C1-C8 alkylene. In embodiments, L5 is independently substituted or unsubstituted C1-C6 alkylene. In embodiments, L5 is independently substituted C1-C6 alkylene. In embodiments, L5 is independently unsubstituted C1-C6 alkylene. In embodiments, L5 is independently substituted or unsubstituted C1-C4 alkylene. In embodiments, L5 is independently substituted C1-C4 alkylene. In embodiments, L5 is independently unsubstituted C1-C4 alkylene. In embodiments, L5 is independently substituted or unsubstituted ethylene. In embodiments, L5 is independently substituted ethylene. In embodiments, L5 is independently unsubstituted ethylene. In embodiments, L5 is independently substituted or unsubstituted methylene. In embodiments, L5 is independently substituted methylene. In embodiments, L5 is independently unsubstituted methylene.
In embodiments, L5 is independently substituted or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In embodiments, L5 is independently substituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In embodiments, L5 is independently unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered). In embodiments, L5 is independently substituted or unsubstituted 2 to 20 membered heteroalkylene. In embodiments, L5 is independently substituted 2 to 20 membered heteroalkylene. In embodiments, L5 is independently unsubstituted 2 to 20 membered heteroalkylene. In embodiments, L5 is independently substituted or unsubstituted 2 to 8 membered heteroalkylene. In embodiments, L5 is independently substituted 2 to 8 membered heteroalkylene. In embodiments, L5 is independently unsubstituted 2 to 8 membered heteroalkylene. In embodiments, L5 is independently substituted or unsubstituted 2 to 6 membered heteroalkylene. In embodiments, L5 is independently substituted 2 to 6 membered heteroalkylene. In embodiments, L5 is independently unsubstituted 2 to 6 membered heteroalkylene. In embodiments, L5 is independently substituted or unsubstituted 4 to 6 membered heteroalkylene. In embodiments, L5 is independently substituted 4 to 6 membered heteroalkylene. In embodiments, L5 is independently unsubstituted 4 to 6 membered heteroalkylene. In embodiments, L5 is independently substituted or unsubstituted 2 to 3 membered heteroalkylene. In embodiments, L5 is independently substituted 2 to 3 membered heteroalkylene. In embodiments, L5 is independently unsubstituted 2 to 3 membered heteroalkylene. In embodiments, L5 is independently substituted or unsubstituted 4 to 5 membered heteroalkylene. In embodiments, L5 is independently substituted 4 to 5 membered heteroalkylene. In embodiments, L5 is independently unsubstituted 4 to 5 membered heteroalkylene.
In embodiments, L5A is independently a bond or unsubstituted alkylene; L5B is independently a bond, —NHC(O)—, or unsubstituted arylene; L5C is independently a bond, unsubstituted alkylene, or unsubstituted arylene; L5D is independently a bond or unsubstituted alkylene; and L5E is independently a bond or —NHC(O)—. In embodiments, L5A is independently a bond or unsubstituted alkylene. In embodiments, L5B is independently a bond, —NHC(O)—, or unsubstituted arylene. In embodiments, L5C is independently a bond, unsubstituted alkylene, or unsubstituted arylene. In embodiments, L5D is independently a bond or unsubstituted alkylene. In embodiments, L5E is independently a bond or —NHC(O)—.
In embodiments, L5A is independently a bond or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L5A is independently unsubstituted C1-C20 alkylene. In embodiments, L5A is independently unsubstituted C1-C12 alkylene. In embodiments, L5A is independently unsubstituted C1-C8 alkylene. In embodiments, L5A is independently unsubstituted C1-C6 alkylene. In embodiments, L5A is independently unsubstituted C1-C4 alkylene. In embodiments, L5A is independently unsubstituted ethylene.
In embodiments, L5A is independently unsubstituted methylene. In embodiments, L5A is independently a bond.
In embodiments, L5B is independently a bond. In embodiments, L5B is independently —NHC(O)—. In embodiments, L5B is independently unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl). In embodiments, L5B is independently unsubstituted C6-C12 arylene. In embodiments, L5B is independently unsubstituted C6-C10 arylene. In embodiments, L5B is independently unsubstituted phenylene. In embodiments, L5B is independently unsubstituted naphthylene.
In embodiments, L5C is independently a bond or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L5C is independently unsubstituted C1-C20 alkylene. In embodiments, L5C is independently unsubstituted C1-C12 alkylene. In embodiments, L5C is independently unsubstituted C1-C8 alkylene. L5C is independently unsubstituted C2-C8 alkynylene. In embodiments, L5C is independently unsubstituted C1-C6 alkylene. In embodiments, L5C is independently unsubstituted C1-C4 alkylene. In embodiments, L5C is independently unsubstituted ethylene. In embodiments, L5C is independently unsubstituted methylene. In embodiments, L5C is independently a bond or unsubstituted alkynylene (e.g., C2-C20, C2-C12, C2-C8, C2-C6, C2-C4, or C2-C2). In embodiments, L5C is independently unsubstituted C2-C20 alkynylene. In embodiments, L5C is independently unsubstituted C2-C12 alkynylene. In embodiments, L5C is independently unsubstituted C2-C8 alkynylene. In embodiments, L5C is independently unsubstituted C2-C6 alkynylene. In embodiments, L5C is independently unsubstituted C2-C4 alkynylene. In embodiments, L5C is independently unsubstituted ethynylene. In embodiments, L5C is independently unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl). In embodiments, L5C is independently unsubstituted C6-C12 arylene. In embodiments, L5C is independently unsubstituted C6-C10 arylene. In embodiments, L5C is independently unsubstituted phenylene.
In embodiments, L5C is independently unsubstituted naphthylene. In embodiments, L5C is independently a bond.
In embodiments, L5D is independently a bond or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L5D is independently unsubstituted C1-C20 alkylene. In embodiments, L5D is independently unsubstituted C1-C12 alkylene. In embodiments, L5A is independently unsubstituted C1-C8 alkylene. In embodiments, L5D is independently unsubstituted C1-C6 alkylene. In embodiments, L5D is independently unsubstituted C1-C4 alkylene. In embodiments, L5D is independently unsubstituted ethylene. In embodiments, L5D is independently unsubstituted methylene. In embodiments, L5D is independently a bond.
In embodiments, L5E is independently a bond. In embodiments, L5E is independently —NHC(O)—.
In embodiments, L5A is independently a bond or unsubstituted C1-C8 alkylene. In embodiments, L5B is independently a bond, —NHC(O)—, or unsubstituted phenylene. In embodiments, L5C is independently a bond, unsubstituted C2-C8 alkynylene, or unsubstituted phenylene. In embodiments, L5D is independently a bond or unsubstituted C1-C8 alkylene. In embodiments, L5E is independently a bond or —NHC(O)—.
In embodiments, L5 is independently a bond,
In embodiments, L5 is independently a bond. In embodiments, L5 is independently
In embodiments L5 is independently
In embodiments, L5 is independently
In embodiments, L5 is independently
In embodiments, L5 is independently
In embodiments, R1 is unsubstituted alkyl (e.g., C1-C25, C1-C20, C1-C17, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R1 is unsubstituted unbranched alkyl (e.g., C1-C25, C1-C20, C1-C17, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R1 is unsubstituted unbranched saturated alkyl (e.g., C1-C25, C1-C20, C1-C17, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R1 is unsubstituted unbranched unsaturated alkyl (e.g., C1-C25, C1-C20, C1-C17, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2).
In embodiments, R1 is unsubstituted C1-C17 alkyl. In embodiments, R1 is unsubstituted C11-C17 alkyl. In embodiments, R1 is unsubstituted C13-C17 alkyl. In embodiments, R1 is unsubstituted C14-C15 alkyl. In embodiments, R1 is unsubstituted C15 alkyl. In embodiments, R1 is unsubstituted C14 alkyl.
In embodiments, R1 is unsubstituted unbranched C1-C17 alkyl. In embodiments, R1 is unsubstituted unbranched C11-C17 alkyl. In embodiments, R1 is unsubstituted unbranched C13-C17 alkyl. In embodiments, R1 is unsubstituted unbranched C14-C15 alkyl. In embodiments, R1 is unsubstituted unbranched C14 alkyl. In embodiments, R1 is unsubstituted unbranched C15 alkyl.
In embodiments, R1 is unsubstituted unbranched saturated C1-C17 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C11-C17 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C13-C17 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C14-C15 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C14 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C15 alkyl.
In embodiments, R1 is unsubstituted unbranched unsaturated C1-C17 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C11-C17 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C13-C17 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C14-C15 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C14 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C15 alkyl.
In embodiments, R2 is unsubstituted alkyl (e.g., C1-C25, C1-C20, C1-C17, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R2 is unsubstituted unbranched alkyl (e.g., C1-C25, C1-C20, C1-C17, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R2 is unsubstituted unbranched saturated alkyl (e.g., C1-C25, C1-C20, C1-C17, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2).
In embodiments, R2 is unsubstituted unbranched unsaturated alkyl (e.g., C1-C25, C1-C20, C1-C17, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2).
In embodiments, R2 is unsubstituted C1-C17 alkyl. In embodiments, R2 is unsubstituted C11-C17 alkyl. In embodiments, R2 is unsubstituted C13-C17 alkyl. In embodiments, R2 is unsubstituted C14-C15 alkyl. In embodiments, R2 is unsubstituted C14 alkyl. In embodiments, R2 is unsubstituted C15 alkyl.
In embodiments, R2 is unsubstituted unbranched C1-C17 alkyl. In embodiments, R2 is unsubstituted unbranched C11-C17 alkyl. In embodiments, R2 is unsubstituted unbranched C13-C17 alkyl. In embodiments, R2 is unsubstituted unbranched C14-C15 alkyl. In embodiments, R2 is unsubstituted unbranched C14 alkyl. In embodiments, R2 is unsubstituted unbranched C15 alkyl.
In embodiments, R2 is unsubstituted unbranched saturated C1-C17 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C11-C17 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C13-C17 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C14-C15 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C14 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C15 alkyl.
In embodiments, R2 is unsubstituted unbranched unsaturated C1-C17 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C11-C17 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C13-C17 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C14-C15 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C14 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C15 alkyl.
In embodiments, at least one of R1 and R2 is unsubstituted C1-C19 alkyl. In embodiments, at least one of R1 and R2 is unsubstituted C9-C19 alkyl. In embodiments, at least one of R1 and R2 is unsubstituted C11-C19 alkyl. In embodiments, at least one of R1 and R2 is unsubstituted C13-C19 alkyl.
In embodiments, R1 is unsubstituted C1-C19 alkyl. In embodiments, R1 is unsubstituted C9-C19 alkyl. In embodiments, R1 is unsubstituted C11-C19 alkyl. In embodiments, R1 is unsubstituted C13-C19 alkyl. In embodiments, R1 is unsubstituted unbranched C1-C19 alkyl. In embodiments, R1 is unsubstituted unbranched C9-C19 alkyl. In embodiments, R1 is unsubstituted unbranched C11-C19 alkyl. In embodiments, R1 is unsubstituted unbranched C13-C19 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C1-C19 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C9-C19 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C11-C19 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C13-C19 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C1-C19 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C9-C19 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C11-C19 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C13-C19 alkyl.
In embodiments, R2 is unsubstituted C1-C19 alkyl. In embodiments, R2 is unsubstituted C9-C19 alkyl. In embodiments, R2 is unsubstituted C11-C19 alkyl. In embodiments, R2 is unsubstituted C13-C19 alkyl. In embodiments, R2 is unsubstituted unbranched C1-C19 alkyl. In embodiments, R2 is unsubstituted unbranched C9-C19 alkyl. In embodiments, R2 is unsubstituted unbranched C11-C19 alkyl. In embodiments, R2 is unsubstituted unbranched C13-C19 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C1-C19 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C9-C19 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C11-C19 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C13-C19 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C1-C19 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C9-C19 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C11-C19 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C13-C19 alkyl.
L1A is independently a bond, —N(R20)—, —O—, —S—, —C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —N(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—O—, —P(S)(NR20R21)—O—, —S—S—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L1A is independently a bond, —N(R20)—, —O—, —S—, —C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —N(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—, —P(S)(NR20R21)—O—, —S—S—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).
In embodiments, L1A is independently a bond, —N(R20)—, —O—, —S—, —C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —N(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—, —P(S)(NR20R21)—O—, —S—S—, unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L1A is substituted, L1A is substituted with a substituent group. In embodiments, when L1A is substituted, L1A is substituted with a size-limited substituent group. In embodiments, when L1A is substituted, L1A is substituted with a lower substituent group.
L1B is independently a bond, —N(R20)—, —O—, —S—, —C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —N(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—O—, —P(S)(NR20R21)—O—, —S—S—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L1B is independently a bond, —N(R20)—, —O—, —S—, —C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —N(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—, —P(S)(NR20R21)—O—, —S—S—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).
In embodiments, L1B is independently a bond, —N(R20)—, —O—, —S—, —C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —N(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—O—, —P(S)(NR20R21)—O—, —S—S—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L1B is substituted, L1B is substituted with a substituent group. In embodiments, when L1B is substituted, L1B is substituted with a size-limited substituent group. In embodiments, when L1B is substituted, L1B is substituted with a lower substituent group.
L1C is independently a bond, —N(R20)—, —O—, —S—, —C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —N(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—O—, —P(S)(NR20R21)—O—, —S—S—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L1C is independently a bond, —N(R20)—, —O—, —S—, —C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —N(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—O—, —P(S)(NR20R21)—O—, —S—S—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).
In embodiments, L1C is independently a bond, N(R20)—, —O—, —S—, —C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —N(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—O—, —P(S)(NR20R21)—O—, —S—S—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L1C is substituted, L1C is substituted with a substituent group. In embodiments, when L1C is substituted, L1C is substituted with a size-limited substituent group. In embodiments, when L1C is substituted, L1C is substituted with a lower substituent group.
R1C is independently substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkyl (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkyl (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkyl (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkyl (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted aryl (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroaryl (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, R1C is independently substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkyl (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkyl (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkyl (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkyl (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) aryl (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroaryl (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, R1C is independently unsubstituted alkyl (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkyl (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkyl (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkyl (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted aryl (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroaryl (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when R1C is substituted, R1C is substituted with a substituent group. In embodiments, when R1C is substituted, R1C is substituted with a size-limited substituent group. In embodiments, when R1C is substituted, R1C is substituted with a lower substituent group. In embodiments, R1C is substituted with oxo (═O).
L1D is independently a bond, —N(R20)—, —O—, —S—, —C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —N(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—O—, —P(S)(NR20R21)—O—, —S—S—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L1D is independently a bond, —N(R20)—, —O—, —S—, —C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—O—, —P(S)(NR20R21)—O—, —S—S—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).
In embodiments, L1D is independently a bond, —N(R20)—, —O—, —S—, —C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —N(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—O—, —P(S)(NR20R21)—O—, —S—S—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L1D is substituted, L1D is substituted with a substituent group. In embodiments, when L1D is substituted, L1D is substituted with a size-limited substituent group. In embodiments, when L1D is substituted, L1D is substituted with a lower substituent group.
R1D is independently substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkyl (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkyl (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkyl (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkyl (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted aryl (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroaryl (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, R1D is independently substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkyl (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkyl (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkyl (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkyl (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) aryl (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroaryl (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, R1D is independently unsubstituted alkyl (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkyl (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkyl (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkyl (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted aryl (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroaryl (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when R1D is substituted, R1D is substituted with a substituent group. In embodiments, when R1D is substituted, R1D is substituted with a size-limited substituent group. In embodiments, when R1D is substituted, R1D is substituted with a lower substituent group.
L1E is independently a bond, —N(R20)—, —O—, —S—, —C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —N(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—O—, —P(S)(NR20R21)—O—, —S—S—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L1E is independently a bond, —N(R20)—, —O—, —S—, C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —N(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—, —P(S)(NR20R21)—O—, —S—S—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered).
In embodiments, L1E is independently a bond, —N(R20)—, —O—, —S—, —C(O)—, —N(R20)C(O)—, —C(O)N(R21)—, —N(R20)C(O)N(R21)—, —C(O)O—, —OC(O)—, —N(R20)C(O)O—, —OC(O)N(R21)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R22)—O—, —O—P(S)(R22)—O—, —O—P(O)(NR20R21)—N—, —O—P(S)(NR20R21)—N—, —O—P(O)(NR20R21)—O—, —O—P(S)(NR20R21)—O—, —P(O)(NR20R21)—N—, —P(S)(NR20R21)—N—, —P(O)(NR20R21)—O—, —P(S)(NR20R21)—O—, —S—S—, unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L1E is substituted, L1E is substituted with a substituent group. In embodiments, when L1E is substituted, L1E is substituted with a size-limited substituent group. In embodiments, when L1E is substituted, L1E is substituted with a lower substituent group.
R1E is independently substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkyl (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkyl (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkyl (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkyl (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted aryl (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroaryl (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, R1E is independently substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkyl (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkyl (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkyl (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkyl (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) aryl (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroaryl (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, R1E is independently unsubstituted alkyl (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkyl (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkyl (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkyl (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted aryl (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroaryl (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when R1E is substituted, R1E is substituted with a substituent group. In embodiments, when R1E is substituted, R1E is substituted with a size-limited substituent group. In embodiments, when R1E is substituted, R1E is substituted with a lower substituent group.
L3 is independently a bond, —N(R23)—, —O—, —S—, —C(O)—, —N(R23)C(O)—, —C(O)N(R24)—, —N(R23)C(O)N(R24)—, —C(O)O—, —OC(O)—, —N(R23)C(O)O—, —OC(O)N(R24)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R25)—O—, —O—P(S)(R25)—O—, —O—P(O)(NR23R24)—N—, —O—P(S)(NR23R24)—N—, —O—P(O)(NR23R24)—O—, —O—P(S)(NR23R24)—O—, —P(O)(NR23R24)—N—, —P(S)(NR23R24)—N—, —P(O)(NR23R24)—O—, —P(S)(NR23R24)—O—, —S—S—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L3 is independently a bond, a —N(R23)—, —O—, —S—, —C(O)—, —N(R23)C(O)—, —C(O)N(R24)—, —N(R23)C(O)N(R24)—, —C(O)O—, —OC(O)—, —N(R23)C(O)O—, —OC(O)N(R24)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R25)—O—, —O—P(S)(R25)—O—, —O—P(O)(NR23R24)—N—, —O—P(S)(NR23R24)—N—, —O—P(O)(NR23 (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L3 is independently a bond, —N(R23)—, —O—, —S—, —C(O)—, —N(R23)C(O)—, —C(O)N(R24)—, —N(R23)C(O)N(R24)—, —C(O)O—, —OC(O)—, —N(R23)C(O)O—, —OC(O)N(R24)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R25)—O—, —O—P(S)(R25)—O—, —O—P(O)(NR23R24)—N—, —O—P(S)(NR23R24)—N—, —O—P(O)(NR23R24)—O—, —O—P(S)(NR23R24)—O—, —P(O)(NR23R24)—N—, —P(S)(NR23R24)—N—, —P(O)(NR23R24)—O—, —P(S)(NR23R24)—O—, —S—S—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L3 is substituted, L3 is substituted with a substituent group. In embodiments, when L3 is substituted, L3 is substituted with a size-limited substituent group. In embodiments, when L3 is substituted, L3 is substituted with a lower substituent group.
L4 is independently a bond, —N(R23)—, —O—, —S—, —C(O)—, —N(R23)C(O)—, —C(O)N(R24)_, —N(R23)C(O)N(R24)—, —C(O)O—, —OC(O)—, —N(R23)C(O)O—, —OC(O)N(R24)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R25)—O—, —O—P(S)(R25)—O—, —O—P(O)(NR23R24)—N—, —O—P(S)(NR23R24)—N—, —O—P(O)(NR23R24)—O—, —O—P(S)(NR23R24)—O—, —P(O)(NR23R24)—N—, —P(S)(NR23R24)—N—, —P(O)(NR23R24)—O—, —P(S)(NR23R24)—O—, —S—S—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L4 is a bond, —N(R23)—, —O—, —S—, —C(O)—, —N(R23)C(O)—, —C(O)N(R24)—, —N(R23)C(O)N(R24)—, —C(O)O—, —OC(O)—, —N(R23)C(O)O—, —OC(O)N(R24)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R25)—O—, —O—P(S)(R25)—O—, —O—P(O)(NR23R24)—N—, —O—P(S)(NR23R24)—N—, —O—P(O)(NR23R24)—O—, —O—P(S)(NR23R24)—O—, —P(O)(NR23R24)—N—, —P(S)(NR23R24)—N—, —P(O)(NR23R24)—O—, —P(S)(NR23R24)—O—, —S—S—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L4 is a bond, —N(R23)—, —O—, —S—, —C(O)—, —N(R23)C(O)—, —C(O)N(R24)—, —N(R23)C(O)N(R24)—, —C(O)O—, —OC(O)—, —N(R23)C(O)O—, —OC(O)N(R24)—, —OPO2—O—, —O—P(O)(S)—O—, —O—P(O)(R25)—O—, —O—P(S)(R25)—O—, —O—P(O)(NR23R24)—N—, —O—P(S)(NR23R24)—N—, —O—P(O)(NR23R24)—O—, —O—P(S)(NR23R24)—O—, —P(O)(NR23R24)—N—, —P(S)(NR23R24)—N—, —P(O)(NR23R24)—O—, —P(S)(NR23R24)—O—, —S—S—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L4 is substituted, L4 is substituted with a substituent group. In embodiments, when L4 is substituted, L4 is substituted with a size-limited substituent group. In embodiments, when L4 is substituted, L4 is substituted with a lower substituent group.
R23 is independently hydrogen or unsubstituted alkyl (e.g., C1-C23, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R23 is independently hydrogen. In embodiments, R23 is independently unsubstituted C1-C23 alkyl. In embodiments, R23 is independently hydrogen or unsubstituted C1-C12 alkyl. In embodiments, R23 is independently hydrogen or unsubstituted C1-C10 alkyl. In embodiments, R23 is independently hydrogen or unsubstituted C1-C8 alkyl. In embodiments, R23 is independently hydrogen or unsubstituted C1-C6 alkyl. In embodiments, R23 is independently hydrogen or unsubstituted C1-C4 alkyl. In embodiments, R23 is independently hydrogen or unsubstituted C1-C2 alkyl.
R24 is independently hydrogen or unsubstituted alkyl (e.g., C1-C23, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R24 is independently hydrogen. In embodiments, R24 is independently unsubstituted C1-C23 alkyl. In embodiments, R24 is independently hydrogen or unsubstituted C1-C12 alkyl. In embodiments, R24 is independently hydrogen or unsubstituted C1-C10 alkyl. In embodiments, R24 is independently hydrogen or unsubstituted C1-C8 alkyl. In embodiments, R24 is independently hydrogen or unsubstituted C1-C6 alkyl. In embodiments, R24 is independently hydrogen or unsubstituted C1-C4 alkyl. In embodiments, R24 is independently hydrogen or unsubstituted C1-C2 alkyl.
R25 is independently hydrogen or unsubstituted alkyl (e.g., C1-C23, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R25 is independently hydrogen. In embodiments, R25 is independently unsubstituted C1-C23 alkyl. In embodiments, R25 is independently hydrogen or unsubstituted C1-C12 alkyl. In embodiments, R25 is independently hydrogen or unsubstituted C1-C10 alkyl. In embodiments, R25 is independently hydrogen or unsubstituted C1-C8 alkyl. In embodiments, R25 is independently hydrogen or unsubstituted C1-C6 alkyl. In embodiments, R25 is independently hydrogen or unsubstituted C1-C4 alkyl. In embodiments, R25 is independently hydrogen or unsubstituted C1-C2 alkyl.
L5 is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L5 is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L5 is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L5 is substituted, L5 is substituted with a substituent group. In embodiments, when L5 is substituted, L5 is substituted with a size-limited substituent group. In embodiments, when L5 is substituted, L5 is substituted with a lower substituent group.
L5A is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L5A is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L5A is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L5A is substituted, L5A is substituted with a substituent group. In embodiments, when L5A is substituted, L5A is substituted with a size-limited substituent group. In embodiments, when L5A is substituted, L5A is substituted with a lower substituent group.
L5B is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L5B is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L5B is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L5B is substituted, L5B is substituted with a substituent group. In embodiments, when L5B is substituted, L5B is substituted with a size-limited substituent group. In embodiments, when L5B is substituted, L5B is substituted with a lower substituent group.
L5C is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L5C is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L5C is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L5C is substituted, L5C is substituted with a substituent group. In embodiments, when L5C is substituted, L5C is substituted with a size-limited substituent group. In embodiments, when L5C is substituted, L5C is substituted with a lower substituent group.
L5D is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L5D is independently a bond, —NH—, —O—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L5D is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L5D is substituted, L5D is substituted with a substituent group. In embodiments, when L5D is substituted, L5D is substituted with a size-limited substituent group. In embodiments, when L5D is substituted, L5D is substituted with a lower substituent group.
L5E is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L5E is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L5E is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L5E is substituted, L5E is substituted with a substituent group. In embodiments, when L5E is substituted, L5E is substituted with a size-limited substituent group. In embodiments, when L5E is substituted, L5E is substituted with a lower substituent group.
L6 is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L6 is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L6 is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L6 is substituted, L6 is substituted with a substituent group. In embodiments, when L6 is substituted, L6 is substituted with a size-limited substituent group. In embodiments, when L6 is substituted, L6 is substituted with a lower substituent group.
L6A is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L61 is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L6A is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L6A is substituted, L6A is substituted with a substituent group. In embodiments, when L6A is substituted, L6A is substituted with a size-limited substituent group. In embodiments, when L6A is substituted, L6A is substituted with a lower substituent group.
L6B is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L6B is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L6B is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L6B is substituted, L6B is substituted with a substituent group. In embodiments, when L6B is substituted, L6B is substituted with a size-limited substituent group. In embodiments, when L6B is substituted, L6B is substituted with a lower substituent group.
L6C is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L6C is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L6C is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L6C is substituted, L6C is substituted with a substituent group. In embodiments, when L6C is substituted, L6C is substituted with a size-limited substituent group. In embodiments, when L6C is substituted, L6C is substituted with a lower substituent group.
L6D is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L6D is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L6D is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L6D is substituted, L6D is substituted with a substituent group. In embodiments, when L6D is substituted, L6D is substituted with a size-limited substituent group. In embodiments, when L6D is substituted, L6D is substituted with a lower substituent group.
L6E is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L6E is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) arylene (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, L6E is independently a bond, —NH—, —O—, —S—, —C(O)—, —NHC(O)—, —NHC(O)NH—, —C(O)O—, —OC(O)—, —C(O)NH—, unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkylene (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkylene (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted arylene (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroarylene (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when L6E is substituted, L6E is substituted with a substituent group. In embodiments, when L6E is substituted, L6E is substituted with a size-limited substituent group. In embodiments, when L6E is substituted, L6E is substituted with a lower substituent group.
In embodiments, L7 is independently substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L7 is independently substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, L7 is independently unsubstituted alkylene (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2).
In embodiments, L7 is independently substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L7 is independently substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L7 is independently unsubstituted heteroalkylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L7 is independently substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkenylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L7 is independently substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkenylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, L7 is independently unsubstituted heteroalkenylene (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 10 membered, 2 to 8 membered, 2 to 6 membered, or 2 to 4 membered). In embodiments, when L7 is substituted, L7 is substituted with a substituent group. In embodiments, when L7 is substituted, L7 is substituted with a size-limited substituent group. In embodiments, when L7 is substituted, L7 is substituted with a lower substituent group.
In embodiments, R1 is unsubstituted alkyl (e.g., C1-C25, C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R1 is unsubstituted C1-C25 alkyl. In embodiments, R1 is unsubstituted C1-C20 alkyl. In embodiments, R1 is unsubstituted C1-C12 alkyl. In embodiments, R1 is unsubstituted C1-C8 alkyl. In embodiments, R1 is unsubstituted C1-C6 alkyl. In embodiments, R1 is unsubstituted C1-C4 alkyl. In embodiments, R1 is unsubstituted C1-C2 alkyl.
In embodiments, R1 is unsubstituted branched alkyl (e.g., C1-C25, C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R1 is unsubstituted branched C1-C25 alkyl.
In embodiments, R1 is unsubstituted branched C1-C20 alkyl. In embodiments, R1 is unsubstituted branched C1-C12 alkyl. In embodiments, R1 is unsubstituted branched C1-C8 alkyl. In embodiments, R1 is unsubstituted branched C1-C6 alkyl. In embodiments, R1 is unsubstituted branched C1-C4 alkyl. In embodiments, R1 is unsubstituted branched C1-C2 alkyl.
In embodiments, R1 is unsubstituted unbranched alkyl (e.g., C1-C25, C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R1 is unsubstituted unbranched C1-C25 alkyl.
In embodiments, R1 is unsubstituted unbranched C1-C20 alkyl. In embodiments, R1 is unsubstituted unbranched C1-C12 alkyl. In embodiments, R1 is unsubstituted unbranched C1-C8 alkyl. In embodiments, R1 is unsubstituted unbranched C1-C6 alkyl. In embodiments, R1 is unsubstituted unbranched C1-C4 alkyl. In embodiments, R1 is unsubstituted unbranched C1-C2 alkyl.
In embodiments, R1 is unsubstituted branched saturated alkyl (e.g., C1-C25, C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R1 is unsubstituted branched saturated C1-C25 alkyl. In embodiments, R1 is unsubstituted branched saturated C1-C20 alkyl.
In embodiments, R1 is unsubstituted branched saturated C1-C12 alkyl. In embodiments, R1 is unsubstituted branched saturated C1-C8 alkyl. In embodiments, R1 is unsubstituted branched saturated C1-C6 alkyl. In embodiments, R1 is unsubstituted branched saturated C1-C4 alkyl. In embodiments, R1 is unsubstituted branched saturated C1-C2 alkyl.
In embodiments, R1 is unsubstituted branched unsaturated alkyl (e.g., C1-C25, C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R1 is unsubstituted branched unsaturated C1-C25 alkyl. In embodiments, R1 is unsubstituted branched unsaturated C1-C20 alkyl. In embodiments, R1 is unsubstituted branched unsaturated C1-C12 alkyl. In embodiments, R1 is unsubstituted branched unsaturated C1-C8 alkyl. In embodiments, R1 is unsubstituted branched unsaturated C1-C6 alkyl. In embodiments, R1 is unsubstituted branched unsaturated C1-C4 alkyl. In embodiments, R1 is unsubstituted branched saturated C1-C2 alkyl.
In embodiments, R1 is unsubstituted unbranched saturated alkyl (e.g., C1-C25, C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R1 is unsubstituted unbranched saturated C1-C25 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C1-C20 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C1-C12 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C1-C8 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C1-C6 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C1-C4 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C1-C2 alkyl.
In embodiments, R1 is unsubstituted unbranched unsaturated alkyl (e.g., C1-C25, C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R1 is unsubstituted unbranched unsaturated C1-C25 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C1-C20 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C1-C12 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C1-C8 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C1-C6 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C1-C4 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C1-C2 alkyl.
In embodiments, R1 is unsubstituted C9-C19 alkyl. In embodiments, R1 is unsubstituted branched C9-C19 alkyl. In embodiments, R1 is unsubstituted unbranched C9-C19 alkyl. In embodiments, R1 is unsubstituted branched saturated C9-C19 alkyl. In embodiments, R1 is unsubstituted branched unsaturated C9-C19 alkyl. In embodiments, R1 is unsubstituted unbranched saturated C9-C19 alkyl. In embodiments, R1 is unsubstituted unbranched unsaturated C9-C19 alkyl.
In embodiments, R2 is unsubstituted alkyl (e.g., C1-C25, C1-C20, C1-C12, C1-C8, C1-C8, C1-C4, or C1-C2). In embodiments, R2 is unsubstituted C1-C25 alkyl. In embodiments, R2 is unsubstituted C1-C20 alkyl. In embodiments, R2 is unsubstituted C1-C12 alkyl. In embodiments, R2 is unsubstituted C1-C8 alkyl. In embodiments, R2 is unsubstituted C1-C6 alkyl. In embodiments, R2 is unsubstituted C1-C4 alkyl. In embodiments, R2 is unsubstituted C1-C2 alkyl.
In embodiments, R2 is unsubstituted branched alkyl (e.g., C1-C25, C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R2 is unsubstituted branched C1-C25 alkyl.
In embodiments, R2 is unsubstituted branched C1-C20 alkyl. In embodiments, R2 is unsubstituted branched C1-C12 alkyl. In embodiments, R2 is unsubstituted branched C1-C8 alkyl. In embodiments, R2 is unsubstituted branched C1-C6 alkyl. In embodiments, R2 is unsubstituted branched C1-C4 alkyl. In embodiments, R2 is unsubstituted branched C1-C2 alkyl.
In embodiments, R2 is unsubstituted unbranched alkyl (e.g., C1-C25, C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R2 is unsubstituted unbranched C1-C25 alkyl.
In embodiments, R2 is unsubstituted unbranched C1-C20 alkyl. In embodiments, R2 is unsubstituted unbranched C1-C12 alkyl. In embodiments, R2 is unsubstituted unbranched C1-C8 alkyl. In embodiments, R2 is unsubstituted unbranched C1-C6 alkyl. In embodiments, R2 is unsubstituted unbranched C1-C4 alkyl. In embodiments, R2 is unsubstituted unbranched C1-C2 alkyl.
In embodiments, R2 is unsubstituted branched saturated alkyl (e.g., C1-C25, C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R2 is unsubstituted branched saturated C1-C25 alkyl. In embodiments, R2 is unsubstituted branched saturated C1-C20 alkyl.
In embodiments, R2 is unsubstituted branched saturated C1-C12 alkyl. In embodiments, R2 is unsubstituted branched saturated C1-C8 alkyl. In embodiments, R2 is unsubstituted branched saturated C1-C6 alkyl. In embodiments, R2 is unsubstituted branched saturated C1-C4 alkyl. In embodiments, R2 is unsubstituted branched saturated C1-C2 alkyl.
In embodiments, R2 is unsubstituted branched unsaturated alkyl (e.g., C1-C25, C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R2 is unsubstituted branched unsaturated C1-C25 alkyl. In embodiments, R2 is unsubstituted branched unsaturated C1-C20 alkyl. In embodiments, R2 is unsubstituted branched unsaturated C1-C12 alkyl. In embodiments, R2 is unsubstituted branched unsaturated C1-C8 alkyl. In embodiments, R2 is unsubstituted branched unsaturated C1-C6 alkyl. In embodiments, R2 is unsubstituted branched unsaturated C1-C4 alkyl. In embodiments, R2 is unsubstituted branched saturated C1-C2 alkyl.
In embodiments, R2 is unsubstituted unbranched saturated alkyl (e.g., C1-C25, C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R2 is unsubstituted unbranched saturated C1-C25 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C1-C20 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C1-C12 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C1-C8 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C1-C6 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C1-C4 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C1-C2 alkyl.
In embodiments, R2 is unsubstituted unbranched unsaturated alkyl (e.g., C1-C25, C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2). In embodiments, R2 is unsubstituted unbranched unsaturated C1-C25 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C1-C20 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C1-C12 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C1-C8 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C1-C6 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C1-C4 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C1-C2 alkyl.
In embodiments, R2 is unsubstituted C9-C19 alkyl. In embodiments, R2 is unsubstituted branched C9-C19 alkyl. In embodiments, R2 is unsubstituted unbranched C9-C19 alkyl. In embodiments, R2 is unsubstituted branched saturated C9-C19 alkyl. In embodiments, R2 is unsubstituted branched unsaturated C9-C19 alkyl. In embodiments, R2 is unsubstituted unbranched saturated C9-C19 alkyl. In embodiments, R2 is unsubstituted unbranched unsaturated C9-C19 alkyl.
In embodiments, R3 is hydrogen, —NH2, —OH, —SH, —C(O)H, —C(O)NH2, —NHC(O)H, —NHC(O)OH, —NHC(O)NH2, —C(O)OH, —OC(O)H, —N3, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted alkyl (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroalkyl (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted cycloalkyl (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heterocycloalkyl (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted aryl (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) or unsubstituted heteroaryl (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, R3 is hydrogen, —NH2, —OH, —SH, —C(O)H, —C(O)NH2, —NHC(O)H, —NHC(O)OH, —NHC(O)NH2, —C(O)OH, —OC(O)H, —N3, substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) alkyl (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroalkyl (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) cycloalkyl (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heterocycloalkyl (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) aryl (e.g., C6-C12, C6-C10, or phenyl), or substituted (e.g., substituted with a substituent group, a size-limited substituent group, or lower substituent group) heteroaryl (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, R3 is hydrogen, —NH2, —OH, —SH, —C(O)H, —C(O)NH2, —NHC(O)H, —NHC(O)OH, —NHC(O)NH2, —C(O)OH, —OC(O)H, —N3, unsubstituted alkyl (e.g., C1-C20, C1-C12, C1-C8, C1-C6, C1-C4, or C1-C2), unsubstituted heteroalkyl (e.g., 2 to 20 membered, 2 to 12 membered, 2 to 8 membered, 2 to 6 membered, 4 to 6 membered, 2 to 3 membered, or 4 to 5 membered), unsubstituted cycloalkyl (e.g., C3-C10, C3-C8, C3-C6, C4-C6, or C5-C6), unsubstituted heterocycloalkyl (e.g., 3 to 10 membered, 3 to 8 membered, 3 to 6 membered, 4 to 6 membered, 4 to 5 membered, or 5 to 6 membered), unsubstituted aryl (e.g., C6-C12, C6-C10, or phenyl), or unsubstituted heteroaryl (e.g., 5 to 12 membered, 5 to 10 membered, 5 to 9 membered, or 5 to 6 membered). In embodiments, when R3 is substituted, R3 is substituted with a substituent group. In embodiments, when R3 is substituted, R3 is substituted with a size-limited substituent group. In embodiments, when R3 is substituted, R3 is substituted with a lower substituent group (e.g., oxo).
In embodiments, the uptake motif is represented by the structure:
The uptake motif is attached to the remainder of the compounds provided here through the -L3-L4- moiety as set forth in Formula (I) above. The wavy line represents attachment to the L4 linker in Formula (I). R1, R2, R3, L5, and L6 in Formula (I-a) are as described in Formula (I), including embodiments thereof.
In embodiments, the compound comprises one or more uptake motifs having a structure shown in Table 2 below. In embodiments, the compound comprises a DTx-01-01 motif in Table 2. In embodiments, the compound comprises a DTx-01-03 motif 1 of Table 2.
In embodiments, the compound comprises a DTx-01-06 motif in Table 2. In embodiments, the compound comprises a DTx-01-08 motif in Table 2. In embodiments, the compound comprises a DTx-01-11 motif in Table 2. In embodiments, the compound comprises a DTx-01-13 motif in Table 2. In embodiments, the compound comprises a DTx-01-30 motif in Table 2. In embodiments, the compound comprises a DTx-01-31 motif in Table 2. In embodiments, the compound comprises a DTx-01-32 motif in Table 2. In embodiments, the compound comprises a DTx-01-33 motif in Table 2. In embodiments, the compound comprises a DTx-01-34 motif in Table 2. In embodiments, the compound comprises a DTx-01-35 motif in Table 2. In embodiments, the compound comprises a DTx-01-36 motif in Table 2. In embodiments, the compound comprises a DTx-01-39 motif in Table 2. In embodiments, the compound comprises a DTx-01-43 motif in Table 2. In embodiments, the compound comprises a DTx-01-44 motif in Table 2. In embodiments, the compound comprises a DTx-01-45 motif in Table 2. In embodiments, the compound comprises a DTx-01-46 motif in Table 2. In embodiments, the compound comprises a DTx-01-50 motif in Table 2. In embodiments, the compound comprises a DTx-01-51 motif in Table 2. In embodiments, the compound comprises a DTx-01-52 motif in Table 2. In embodiments, the compound comprises a DTx-01-53 motif in Table 2. In embodiments, the compound comprises a DTx-01-54 motif in Table 2. In embodiments, the compound comprises a DTx-01-55 motif in Table 2. In embodiments, the compound comprises a DTx-03-06 motif in Table 2. In embodiments, the compound comprises a DTx-03-50 motif in Table 2. In embodiments, the compound comprises a DTx-03-51 motif in Table 2. In embodiments, the compound comprises a DTx-03-52 motif in Table 2. In embodiments, the compound comprises a DTx-03-53 motif in Table 2. In embodiments, the compound comprises a DTx-03-54 motif in Table 2. In embodiments, the compound comprises a DTx-03-55 motif in Table 2. In embodiments, the compound comprises a DTx-04-01 motif in Table 2. In embodiments, the compound comprises a DTx-05-01 motif in Table 2. In embodiments, the compound comprises a DTx-06-06 motif in Table 2. In embodiments, the compound comprises a DTx-06-50 motif in Table 2. In embodiments, the compound comprises a DTx-06-51 motif in Table 2. In embodiments, the compound comprises a DTx-06-52 motif in Table 2. In embodiments, the compound comprises a DTx-06-53 motif in Table 2. In embodiments, the compound comprises a DTx-06-54 motif in Table 2. In embodiments, the compound comprises a DTx-06-55 motif in Table 2. In embodiments, the compound comprises a DTx-08-01 motif in Table 2. In embodiments, the compound comprises a DTx-09-01 motif in Table 2. In embodiments, the compound comprises a DTx-10-01 motif in Table 2. In embodiments, the compound comprises a DTx-11-01 motif in Table 2. In embodiments, the compound comprises a DTx-01-60 motif in Table 2. In embodiments, the compound comprises a DTx-01-61 motif in Table 2. In embodiments, the compound comprises a DTx-01-62 motif in Table 2. In embodiments, the compound comprises a DTx-01-63 motif in Table 2. In embodiments, the compound comprises a DTx-01-64 motif in Table 2. In embodiments, the compound comprises a DTx-01-65 motif in Table 2. In embodiments, the compound comprises a DTx-01-66 motif in Table 2. In embodiments, the compound comprises a DTx-01-67 motif in Table 2. In embodiments, the compound comprises a DTx-01-68 motif in Table 2. In embodiments, the compound comprises a DTx-01-69 motif in Table 2. In embodiments, the compound comprises a DTx-01-70 motif in Table 2. In embodiments, the compound comprises a DTx-01-71 motif in Table 2. In embodiments, the compound comprises a DTx-01-72 motif in Table 2. In embodiments, the compound comprises a DTx-01-73 motif in Table 2. In embodiments, the compound comprises a DTx-01-74 motif in Table 2. In embodiments, the compound comprises a DTx-01-75 motif in Table 2. In embodiments, the compound comprises a DTx-01-76 motif in Table 2. In embodiments, the compound comprises a DTx-01-77 motif in Table 2. In embodiments, the compound comprises a DTx-01-78 motif in Table 2. In embodiments, the compound comprises a DTx-01-79 motif in Table 2. In embodiments, the compound comprises a DTx-01-80 motif in Table 2. In embodiments, the compound comprises a DTx-01-81 motif in Table 2. In embodiments, the compound comprises a DTx-01-82 motif in Table 2. In embodiments, the compound comprises a DTx-1-83 motif in Table 2. In embodiments, the compound comprises a DTx-01-84 motif in Table 2. In embodiments, the compound comprises a DTx-01-85 motif in Table 2. In embodiments, the compound comprises a DTx-01-86 motif in Table 2. In embodiments, the compound comprises a DTx-01-87 motif in Table 2. In embodiments, the compound comprises a DTx-01-88 motif in Table 2. In embodiments, the compound comprises a DTx-01-89 motif in Table 2. In embodiments, the compound comprises a DTx-01-90 motif in Table 2. In embodiments, the compound comprises a DTx-01-91 motif in Table 2. In embodiments, the compound comprises a DTx-01-92 motif in Table 2. In embodiments, the compound comprises a DTx-01-93 motif in Table 2. In embodiments, the compound comprises a DTx-01-94 motif in Table 2. In embodiments, the compound comprises a DTx-01-95 motif in Table 2. In embodiments, the compound comprises a DTx-01-96 motif in Table 2. In embodiments, the compound comprises a DTx-01-97 motif in Table 2. In embodiments, the compound comprises a DTx-01-98 motif in Table 2. In embodiments, the compound comprises a DTx-01-99 motif in Table 2. In embodiments, the compound comprises a DTx-01-100 motif in Table 2. In embodiments, the compound comprises a DTx-01-101 motif in Table 2.
In embodiments, DTx-01-01 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-03 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-06 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-08 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-11 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-13 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-30 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-31 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-32 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-33 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-34 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-35 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-36 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-39 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-43 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-44 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-45 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-46 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-50 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-51 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-52 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-53 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-54 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-55 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-03-06 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-03-50 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-03-51 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-03-52 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-03-53 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-03-54 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-03-55 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-04-01 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-05-01 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-06-06 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-06-50 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-06-51 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-06-52 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-06-53 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-06-54 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-06-55 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-08-01 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-09-01 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-10-01 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-11-01 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-60 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-61 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-62 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-63 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-64 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-65 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-66 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-67 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-68 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-69 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-70 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-71 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-72 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-73 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-74 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-75 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-76 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-77 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-78 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-79 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-80 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-81 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-82 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-83 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-84 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-85 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-86 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-87 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-88 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-89 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-90 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-91 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-92 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-93 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-94 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-95 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-96 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-97 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-98 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-99 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-100 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-101 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-01 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-03 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-06 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-08 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-11 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-13 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-30 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-31 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-32 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-33 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-34 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-35 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-36 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-39 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-43 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-44 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-45 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-46 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-50 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-51 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-52 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-53 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-54 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-55 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
n embodiments, DTx-03-06 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-03-50 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-03-51 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-03-52 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-03-53 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-03-54 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-03-55 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-04-01 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-05-01 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-06-06 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-06-50 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-06-51 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-06-52 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-06-53 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-06-54 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-06-55 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-08-01 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-09-01 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-10-01 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-11-01 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-60 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-61 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-62 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-63 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-64 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-65 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-66 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-67 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-68 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-69 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-70 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-71 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-72 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-73 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-74 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-75 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-76 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-77 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-78 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-79 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-80 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-81 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-82 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-83 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-84 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-85 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-86 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-87 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-88 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-89 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-90 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-91 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-92 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-93 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-94 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-95 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-96 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-97 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-98 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-99 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-100 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, DTx-01-101 is attached to the double-stranded nucleic acid (A) through -L3-L4-, wherein -L3-L4- is
In embodiments, -L3-L4- is
the phosphate group is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, and R2 is unsubstituted unbranched C15 alkyl.
In embodiments, -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C13 alkyl, and R2 is unsubstituted unbranched C13 alkyl.
In embodiments, -L3-L4- is
within -L3-L4-, -L3 is attached to a phosphate group at the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
H L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, and R2 is unsubstituted unbranched C15 alkyl.
In embodiments, -L3-L4- is
within -L3-L4-, -L3 is attached to a phosphate group at the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
H, L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C13 alkyl, and R2 is unsubstituted unbranched C13 alkyl.
In embodiments, a compound is DT-000623, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-OH—UFSCMSCFUMGFUMUFGMCFUMGFAMGFUMAFUMCFSAMSUF-3′ (SEQ ID NO: 652), and the nucleotide sequence of the antisense strand is
5′-PO4-AMSUFSGMAFUMAFCMUFCMAFGMCFAMAFCMAFGMGFAMSTDSTD-OH-3′ (SEQ ID NO: 176), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a nucleotide followed by a subscript “D” is a beta-D-deoxyribonucleotide; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, a compound is DT-000812, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-OH—CFSCMSUFCMCFUMGFUMUFGMCFUMGFAMGFUMAFUMCFSAMSUF-3′ (SEQ ID NO: 658), and the nucleotide sequence of the antisense strand is
5′-VP-AMSUFSGMAFUMAFCMUFCMAFGMCFAMAFCMAFGMGFAMGFGMSAMSGM-OH-3′ (SEQ ID NO: 879), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-VP” is a 5′-vinylphosphonate at the 5′-terminal nucleotide of the antisense strand. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, a compound is DT-001246, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-OH—CFSCMSUFCMCFUMGFUMUFGMCFUFGFAMGFUMAFUMCFSAMSUF-3′ (SEQ ID NO: 770), and the nucleotide sequence of the antisense strand is
5′-VP-AMSUFSGMAFUMAFCMUFCMAMGMCFAMAFCMAFGMGFAMGFGMSAMSGM-OH-3′ (SEQ ID NO: 899), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-VP” is a 5′-vinylphosphonate at the 5′-terminal nucleotide of the antisense strand. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, a compound is DT-001247, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
H L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-OH—CFSCMSUFCMCFUMGFUMUFGFCFUMGFAMGFUMAFUMCFSAMSUF-3′ (SEQ ID NO: 771), and the nucleotide sequence of the antisense strand is
5′-VP-AMSUFSGMAFUMAFCMUFCMAFGMCMAMAFCMAFGMGFAMGFGMSAMSGM-OH-3′ (SEQ ID NO: 900), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-VP” is a 5′-vinyl phosphonate at the 5′-terminal nucleotide of the antisense strand. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, a compound is DT-001250, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-OH—CMSCMSUMCMCFUMGFUMUFGMCFUMGFAMGFUMAFUMCMSAMSUM-3′ (SEQ ID NO: 772), and the nucleotide sequence of the antisense strand is
5′-VP-AMSUFSGMAFUMAFCMUFCMAFGMCFAMAFCMAFGMGFAMGFGMSAMSGM-OH-3′ (SEQ ID NO: 879), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-VP” is a 5′-VP modification at the 5′-terminal nucleotide of the antisense strand. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, a compound is DT-001251, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-OH—CMSCMSUMCMCMUMGFUMUFGMCFUMGFAMGFUMAFUMCMSAMSUM-3′ (SEQ ID NO: 773), and the nucleotide sequence of the antisense strand is
5′-VP-AMSUFSGMAFUMAFCMUMCMAFGMCMAMAFCMAFGMGFAMGFGMSAMSGM-OH-3′ (SEQ ID NO: 901), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-VP” is a 5′-VP modification at the 5′-terminal nucleotide of the antisense strand. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, a compound is DT-001252, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-OH—CMSCMSUMCMCMUMGFUMUFGFCFUMGMAMGMUMAMUMCMSAMSUM-3′ (SEQ ID NO: 774), and the nucleotide sequence of the antisense strand is
5′-VP-AMSUFSGMAMUMAFCMUMCMAMGMCMAMAFCMAFGMGMAMGMGMSAMSGM-OH-3′ (SEQ ID NO: 902), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-VP” is a 5′-VP modification at the 5′-terminal nucleotide of the antisense strand. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, a compound is DT-001253, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-OH—CMSCMSUMCMCMUMGFUMUFGFCFUMGMAMGMUMAMUMCMAMUM-3′ (SEQ ID NO: 775), and the nucleotide sequence of the antisense strand is
5′-VP-AMSUFSGMAMUMAFCMUMCMAMGMCMAMAFCMAFGMGMAMGMGMSAMSGM-OH-3′ (SEQ ID NO: 902), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-VP” is a 5′-VP modification at the 5′-terminal nucleotide of the antisense strand. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, a compound is DT-001254, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-OH—CESCESUMCMCFUMGFUMUFGMCFUMGFAMGFUMAFUMCMSAMSUM-3′ (SEQ ID NO: 776), and the nucleotide sequence of the antisense strand is
5′-VP-AMSUFSGMAFUMAFCMUFCMAFGMCFAMAFCMAFGMGFAMGFGMSAMSGM-OH-3′ (SEQ ID NO: 879), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a nucleotide followed by the subscript “E” is a 2′-O-methoxyethyl nucleotide; the nucleobase of each “CE” nucleotide is a 5-methylcytosine; each other “C” is a non-methylated cytosine; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-VP” is a 5′-VP modification at the 5′-terminal nucleotide of the antisense strand. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, a compound is DT-001255, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-OH—CMSCESUECMCFUMGFUMUFGMCFUMGFAMGFUMAFUMCMSAMSUM-3′ (SEQ ID NO: 777), and the nucleotide sequence of the antisense strand is
5′-VP-AMSUFSGMAFUMAFCMUFCMAFGMCFAMAFCMAFGMGFAMGFGMSAMSGM-OH-3′ (SEQ ID NO: 879), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a nucleotide followed by the subscript “E” is a 2′-O-methoxyethyl nucleotide; the nucleobase of each “CE” nucleotide is a 5-methylcytosine; each other “C” is a non-methylated cytosine; the nucleobase of each “UE” nucleotide is a 5-methyluracil; each other “U” is a non-methylated uridine; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-VP” is a 5′-VP modification at the 5′-terminal nucleotide of the antisense strand. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, a compound is DT-001256, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-OH—CMSCESUECMCFUMGFUMUFGMCFUMGFAMGFUMAFUMCESAESUM-3′ (SEQ ID NO: 778), and the nucleotide sequence of the antisense strand is
5′-VP-AMSUFSGMAFUMAFCMUFCMAFGMCFAMAFCMAFGMGFAMGFGMSAMSGM-OH-3′ (SEQ ID NO: 879), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a nucleotide followed by the subscript “E” is a 2′-O-methoxyethyl nucleotide; the nucleobase of each “CE” nucleotide is a 5-methylcytosine; each other “C” is a non-methylated cytosine; the nucleobase of each “UE” nucleotide is a 5-methyluracil; each other “U” is a non-methylated uridine; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-VP” is a 5′-VP modification at the 5′-terminal nucleotide of the antisense strand. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, a compound is DT-001257, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-OH—CESCESUECECFUMGFUMUFGMCFUMGFAMGFUMAFUMCMSAMSUM-3′ (SEQ ID NO: 779), and the nucleotide sequence of the antisense strand is
5′-VP-AMSUFSGMAFUMAFCMUFCMAFGMCFAMAFCMAFGMGFAMGFGMSAMSGM-OH-3′ (SEQ ID NO: 879), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a nucleotide followed by the subscript “E” is a 2′-O-methoxyethyl nucleotide; the nucleobase of each “CE” nucleotide is a 5-methylcytosine; each other “C” is a non-methylated cytosine; the nucleobase of each “UE” nucleotide is a 5-methyluracil; each other “U” is a non-methylated uridine; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-VP” is a 5′-VP at the 5′-terminal nucleotide. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, a compound is DT-001858, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-OH—CMSCMSUMCMCMUMGFUMUFGFCFUMGMAMGMUMAMUMCMAMSUM-3′ (SEQ ID NO: 887), and the nucleotide sequence of the antisense strand is
5′-VP-AMSUFSGMAMUMAFCMUMCMAMGMCMAMAFCMAFGMGMAMGMGMSAMSGM-OH-3′ (SEQ ID NO: 902), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-VP” is a 5′-VP at the 5′-terminal nucleotide. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, a compound is DT-001859, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-OH—CMSCMSUFCMCMUMGFUMUFGFCFUMGMAMGMUMAMUMCMSAMSUM-3′ (SEQ ID NO: 878), and the nucleotide sequence of the antisense strand is
5′-VP-AMSUFSGMAMUMAFCMUMCMAMGMCMAMAFCMAFGMGMAMGMGMSAMSGM-OH-3′ (SEQ ID NO: 902), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-VP” is a 5′-VP at the 5′-terminal nucleotide. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, a compound is DT-001860, where -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl, the nucleotide sequence of the sense strand is
5′-HO—CMSCMSUMCMCMUMGFUMUFGFCFUMGMAMGMUMAMUMCMSAMSUM-3′ (SEQ ID NO: 774), and the nucleotide sequence of the antisense strand is
5′-VP-AMSUFSGMAMUMAFCMUMCMAMGMCMAMAFCMAFGMGMAMGMGMSAMSGE-OH-3′ (SEQ ID NO: 975), where a nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a superscript “S” is a phosphorothioate internucleotide linkage; and all other internucleotide linkages are phosphodiester internucleotide linkages. “5′-VP” is a 5′-VP at the 5′-terminal nucleotide. “5′-OH” and “OH-3′” are hydroxyl moieties at the 5′-terminus and 3′ terminus, respectively.
In embodiments, -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl;
In embodiments, -L3-L4- is
the phosphate group of -L3-L4- is attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand, L6 is
L5 is —NHC(O)—, R3 is hydrogen, R1 is unsubstituted unbranched C15 alkyl, R2 is unsubstituted unbranched C15 alkyl;
In embodiments, a ligand is a saturated or unsaturated C5-C20 alkyl. In embodiments, a ligand contains a saturated or unsaturated C6-C18 alkyl.
The compounds provided herein may be present as a pharmaceutical salt. In embodiments, the pharmaceutical salt is a sodium salt.
Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.
Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.
Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like; particularly preferred are the ammonium, potassium, s16 odium, calcium and magnesium salts. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, specifically such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. Many such salts are known in the art, as described in WO 87/05297, Johnston et al., published Sep. 11, 1987 (incorporated by reference herein in its entirety).
In embodiments, a non-bridging heteroatom (e.g., an S− or O−) of a linkage of a compound provided herein may be protonated or associated with a counterion such as Na+, K+, etc. An acceptable salt (e.g. a pharmaceutically acceptable salt) of a compound may comprise fewer cationic counterions (such as Na+, K+, etc.) than there are non-bridging heteroatoms per molecule (i.e., some non-bridging heteroatoms are protonated and some are associated with counterions). In embodiments, a phosphate linkage attaching an -L3-L4- to a carbon of a nucleotide includes a non-bridging heteroatom. In embodiments, a phosphodiester linkage of a nucleic acid includes a non-bridging heteroatom. In embodiments, a phosphorothioate linkage of a nucleic acid includes a non-bridging heteroatom.
The compounds provided herein may be present as a pharmaceutical composition comprising the compound and a pharmaceutically acceptable diluent. In embodiments, the compound is present in a pharmaceutically acceptable diluent. In embodiments, the pharmaceutically acceptable diluent is a sterile aqueous solution. In embodiments, the sterile aqueous solution is a sterile saline solution.
A pharmaceutical composition may be prepared so that it is compatible with the intended mode of administration of the compound. Routes of administration of compounds include intravenous, intradermal, subcutaneous, transdermal, intramuscular, topical, and ocular administration.
Pharmaceutical compositions may be prepared for ocular administration to the eye in the form of an injection. Pharmaceutical compositions suitable for injection include sterile aqueous solutions, including sterile saline solutions. Pharmaceutical compositions suitable for injection may also be a lyophilized compound that is subsequently reconstitute with a pharmaceutically acceptable diluent in preparation for injection.
Alternatively, pharmaceutical compositions may be prepared for ocular administration to the eye in the form of an ophthalmic suspension (i.e. eye drops). Additional pharmaceutical preparations suitable for ocular administration include emulsions, ointments, aqueous gels, nanomicelles, nanoparticles, liposomes, dendrimers, implants, contact lenses, nanosuspensions, microneedles, and in situ thermosensitive gels.
Provided herein is a method for inhibiting the expression of peripheral myelin protein 22 (PMP22) mRNA in a cell, comprising contacting a cell with a nucleic compound provided herein, thereby inhibiting the expression of peripheral myelin protein 22 (PMP22) in the cell. In embodiments, the cell is a peripheral nerve cell. In embodiments, the cell is in vivo. In embodiments, the cell is in vitro.
Provided herein is a method for inhibiting the expression of peripheral myelin protein 22 (PMP22) in a subject, comprising administering to the subject an effective amount of a compound or pharmaceutical composition provided herein. In embodiments, the expression of peripheral myelin protein 22 (PMP22) is inhibited in the subject. In embodiments, the expression of PMP22 mRNA is inhibited in a peripheral nerve of the subject. In embodiments, the peripheral nerve is one or more of a sciatic nerve, a brachial plexus nerve, a tibial nerve, a peroneal nerve, a femoral nerve, a lateral femoral cutaneous nerve, and a spinal accessory nerve.
Provided herein is a method for increasing myelination and/or slowing the loss of myelination in a subject, comprising administering to the subject an effective amount of a compound or pharmaceutical composition provided herein. In embodiments, the administering increases myelination in the subject. In embodiments, the administering slows the loss of myelination in the subject. In embodiments, the subject has a peripheral demyelinating disease. In embodiments, the peripheral demyelinating disease is Charcot-Marie-Tooth disease (CMT). In embodiments, the Charcot-Marie-Tooth disease is Charcot-Marie-Tooth disease Type TA (CMT1A). In embodiments, the Charcot-Marie-Tooth disease Type 1E (CMT1E).
Provided herein is a method for treating Charcot-Marie-Tooth disease (CMT) in a subject in need thereof, comprising administering to the subject an effective amount compound or pharmaceutical composition provided herein. In embodiments, the Charcot-Marie-Tooth disease (CMT) is Charcot-Marie-Tooth disease Type 1A (CMT1A).
Provided herein is a method for treating Charcot-Marie-Tooth disease Type 1A (CMT1A) in a subject in need thereof, comprising administering to the subject an effective amount compound or pharmaceutical composition provided herein. Provided herein is a method for slowing the progression of Charcot-Marie-Tooth Disease Type 1A (CMT1A) in a subject in need thereof, comprising administering to the subject a compound or pharmaceutical composition provided herein.
In embodiments, the subject has Charcot-Marie-Tooth Disease Type 1A (CMT1A). CMT1A may be diagnosed by a medical professional using one or more routinely available assessments, including family history, medical history, and neurological examination. In embodiments, a subject is diagnosed as having CMT1A by the presence of one or more clinical indicators of CMT1A selected from: a family history of CMT1A; amplification of the PMP22 gene; distal muscle weakness; distal musculature atrophy, decreased deep tendon reflexes, distal sensory impairment; decreased compound muscle action potential; and decreased nerve conduction velocity.
Provided herein is a method for delaying the onset of CMT1A in a subject at risk for developing CMT1A, comprising administering to the subject a compound provided herein. A subject at risk for developing CMT1A may be identified by a medical professional using one or more routinely available assessments, including family history, medical history, and neurological examination. In embodiments, a subject is identified as being at risk for developing CMT1A by the presence of one or more clinical indicators of CMT1A selected from: a family history of CMT1A; amplification of the PMP22 gene; distal muscle weakness; distal musculature atrophy; decreased deep tendon reflexes; distal sensory impairment; decreased compound muscle action potential; and decreased nerve conduction velocity.
In embodiments, a subject has a family history of CMT1A. In embodiments, amplification of the PMP22 gene in the subject is confirmed by genetic testing.
In embodiments, a subject has distal muscle weakness. In embodiments, the distal muscle weakness is in one or more of the arms, legs, hands and feet. In embodiments, the distal muscle weakness is measured by quantified muscular testing (QMT). In embodiments, the distal muscle weakness is reduced hand grip strength. In embodiments, the distal muscle weakness is reduced foot dorsiflexion.
In embodiments, a subject has distal musculature atrophy. In embodiments, the distal musculature atrophy is in one or more of the arms, legs, hands, and feet. In embodiments, the distal musculature atrophy is calf muscle atrophy.
In embodiments, a subject has reduced deep tendon reflexes.
In embodiments, a subject has distal sensory impairment.
In embodiments, the subject has reduced nerve conduction velocity (NCV). In embodiments, the nerve conduction velocity is motor nerve conduction velocity (MNCV). In embodiments, the nerve conduction velocity is sensory nerve conduction velocity (SNCV). Nerve conduction velocity may be determined by an electroneuroagraphy, i.e. a nerve conduction study, involving the placement of electrodes on the skin over a muscle or nerve. These electrodes produce a small electric impulse that stimulates nerves and allows for quantification of electrical activity from a distal muscle or nerve (those in the hands, lower arms, lower legs, and feet).
In embodiments, a subject has reduced compound muscle action potential (CMAP). CMAP may be determined by electromyography (EMG), a procedure which involves inserting a needle electrode through the skin to the muscle and measuring the bioelectrical activity of muscles, specific abnormalities in which indicate axon loss. EMG may be useful in further characterizing the distribution, activity, and severity of peripheral nerve involvement in CMT1A.
In embodiments, a subject has increased calf muscle fat fraction. In embodiments, calf muscle fat fraction is measured by magnetic resonance imaging (MRI).
In embodiments, a subject has elevated plasma neurofilament light (NfL) protein. In embodiments, a subject has elevated plasma transmembrane protease serine 5 (TMPRSS55).
In embodiments, the administration of the compound or pharmaceutical composition to the subject improves and/or slows the progression of one or more clinical indicators of Charcot-Marie-Tooth disease Type 1A in the subject. In embodiments, administration of the compound or pharmaceutical composition to the subject improves one or more clinical indicators of Charcot-Marie-Tooth disease Type 1A in the subject. In embodiments, administration of the compound or pharmaceutical composition to the subject slows the progression of one or more clinical indicators of Charcot-Marie-Tooth disease Type 1A in the subject. In embodiments, the one or more clinical indicator is selected from distal muscle weakness; distal sensory impairment; reduced nerve conduction velocity; reduced compound muscle action potential; reduced sensory nerve action potential; increased calf muscle fat fraction; elevated plasma neurofilament light (NfL); and elevated plasma transmembrane protease serine 5 (TMPRSS55). In embodiments, administration of the compound or pharmaceutical composition to the subject improves distal muscle weakness. In embodiments, administration of the compound slows the progression of distal muscle weakness. In embodiments, the distal muscle weakness is reduced hand grip strength. In embodiments, the distal muscle weakness is reduced foot dorsiflexion. In embodiments, administration of the compound or pharmaceutical composition improves distal sensory impairment. In embodiments, administration of the compound or pharmaceutical composition slows the progress of distal sensory impairment. In embodiments, administration of the compound or pharmaceutical composition increases nerve conduction velocity. In embodiments, administration of the compound or pharmaceutical composition slows the progression of reduced nerve conduction velocity. In embodiments, the nerve conduction velocity is motor nerve conduction velocity. In embodiments, the nerve conduction velocity is sensory nerve conduction velocity. In embodiments, administration of the compound or pharmaceutical composition improves compound muscle action potential. In embodiments, administration of the compound slows the progression of reduced compound muscle action potential. In embodiments, administration of the compound or pharmaceutical composition improves sensory nerve action potential. In embodiments, administration of the compound or pharmaceutical composition slows the progression of reduced sensory nerve action potential. In embodiments, administration of the compound or pharmaceutical composition improves increased fat muscle fat fraction. In embodiments, administration of the compound or pharmaceutical composition slows the progression of increased fat muscle fat fraction. In embodiments, administration of the compound or pharmaceutical composition improves elevated plasma neurofilament light (NfL). In embodiments, administration of the compound or pharmaceutical composition slows the progression of elevated plasma neurofilament light (NfL). In embodiments, administration of the compound or pharmaceutical composition improves elevated plasma transmembrane protease serine 5 (TMPRSS55). In embodiments, administration of the compound or pharmaceutical composition slows the progression of elevated plasma transmembrane protease serine 5 (TMPRSS55).
Disease severity and disease progression in subjects may be determined using one or more clinical assessments. In embodiments, disease severity in a subject is determined by performing one or more clinical assessments. In embodiments, disease progression in a subject is determined by performing one or more clinical assessments. In embodiments, disease progression is determined by measuring the change over time in one or more clinical assessments. In embodiments, the clinical assessment is selected from the Charcot-Marie-Tooth Neuropathy Score (CMTNS), the Charcot-Marie-Tooth Neuropathy Score with Rasch weighting (CMTNS-R), the Charcot Marie-Tooth Neuropathy Score Version 2 (CMTNS-v2), the Charcot-Marie-Tooth Examination Score (CMTES), the Charcot-Marie-Tooth Examination Score with Rasch weighting (CMTES-R), the Charcot-Marie-Tooth Functional Outcome Measure (CMT-FOM), the Charcot-Marie-Tooth Disease Pediatric Scale, the Charcot-Marie-Tooth Disease Infant Scale, the Charcot-Marie-Tooth Health Index, and the Overall Neuropathy Limitation Scale (ONLS). In embodiments, the clinical assessment is the Charcot-Marie-Tooth Neuropathy Score (CMTNS). In embodiments, the clinical assessment is the Charcot-Marie-Tooth Neuropathy Score with Rasch weighting (CMTNS-R). In embodiments, the clinical assessment is the Charcot Marie-Tooth Neuropathy Score Version 2 (CMTNS-v2). In embodiments, the clinical assessment is the Charcot-Marie-Tooth Examination Score (CMTES). In embodiments, the clinical assessment is the Charcot-Marie-Tooth Examination Score with Rasch weighting (CMTES-R). In embodiments, the clinical assessment is the Charcot-Marie-Tooth Functional Outcome Measure (CMT-FOM). In embodiments, the clinical assessment is the Charcot-Marie-Tooth Disease Pediatric Scale. In embodiments, the clinical assessment is the Charcot-Marie-Tooth Disease Infant Scale. In embodiments, the clinical assessment the Charcot-Marie-Tooth Health Index. In embodiments, the clinical assessment is and the Overall Neuropathy Limitation Scale (ONLS).
In embodiments, administration is intravenous administration. In embodiments, the administration is subcutaneous administration.
In embodiments, at least one additional therapy is administered to the subject. In embodiments, the at least one additional therapy is PXT3003 comprising baclofen, sorbitol, and naltrexone.
In embodiments, compounds provided herein are for use in therapy. In embodiments, pharmaceutical compositions provided herein are for use in therapy. In embodiments, the therapy is the treatment of a demyelinating disease. In embodiments, the therapy is the treatment of Charcot-Marie-Tooth disease. In embodiments, the therapy is the treatment of Charcot-Marie-Tooth disease Type 1A (CMT1A).
Various formulations are available to facilitate compound use both in vitro and as therapeutic agents. Accordingly, in embodiments, a compound provided herein is present in a formulation.
Compounds may be formulated with cationic lipids to facilitate transfection into cells. Suitable cationic lipid reagents for transfection include Lipofectamine reagents, such as Lipofectamine RNAiMAX.
For use in vivo as therapeutic agents, nucleic acids compounds may be encapsulated into lipid nanoparticles. Lipid nanoparticles generally comprise a cationic lipid, a non-cationic lipid, and a lipid that prevents aggregation of the nanoparticle. Suitable cationic lipids include DLin-MC3-DMA ((6Z,9Z,28Z,31Z)-Heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate), DLin-KC2-DMA
The following examples are presented to more fully illustrate some embodiments of the invention. They should not be construed, however, as limiting the scope of the invention.
Variations of these examples within the scope of the claims are within the purview of one skilled in the art and are considered to fall within the scope of the embodiments as described and claimed herein. The reader will recognize that the skilled artisan, armed with the present disclosure and skill in the art, is able to prepare and use the invention without exhaustive examples.
To a stirred solution of linear fatty acid 01-08-1 (25.58 g, 0.099 mol) in DMF (500 mL) at RT was added DIPEA (42.66 mL, 0.245 mol) and compound 01-08-2 (8.0 g, 0.049 mol), followed by EDCl (18.97 g, 0.099 mol) and HOBt (13.37 g, 0.099 mol). The resulting mixture was stirred at 50° C. After 16 h, the reaction mixture was quenched with ice water and extracted with DCM. The combined organic extract was washed with water, brine, dried over Na2SO4, and then evaporated to give crude 01-08-3, which was recrystallized (20% MTBE in petroleum ether) to afford 01-08-3 as an off-white solid (18 g, 56%).
To a stirred solution of 01-08-3 (10 g, 0.0156 mol) in MeOH and THF (1:1; 200 mL) at RT was added slowly Ba(OH)2 (9.92 g, 0.031 mol, dissolved in MeOH). The resulting mixture was stirred at RT. After 6 h, the reaction mixture was quenched with ice water dropwise, and then acidified with 1.5 M HCl. The mixture was filtered, and the precipitate was recrystallized (MTBE in petroleum ether) to afford lipid motif DTx-01-08 as an off-white solid (7.2 g, 74.2%). MS (ESI) m/z (M+H)+: 623.6; 1H-NMR (400 MHz, CDCl3): δ 0.868 (m, 6H), 1.25-1.69 (m, 58H), 2.03 (t, J=7.2 Hz, 2H), 2.11 (t, J=7.6 Hz, 2H), 2.99 (q, J=8.4 Hz, 2H), 4.15-4.20 (m, 1H), 7.42 (br s, 1H), 7.65 (d, J=7.6 Hz, 1H), 12.09 (br s, 1H).
To a stirred solution of 01-32-2 (3 g, 0.01 mol) in DMF (50 mL) at RT was added slowly DIPEA (13.8 mL, 0.077 mol), linear fatty acid 01-32-1 (4.4 g, 0.0154 mol), and HATU (5.87 g, 0.0154 mol). The resulting mixture was stirred at 60° C. After 16 h, the reaction mixture was quenched with ice water, the solids isolated by filtration, and the solids dried under vacuum to afford 01-32-3 as an off-white solid (3.5 g, 53.2%).
To a stirred solution of 01-32-3 (3.5 g, 0.0051 mol) in MeOH (10 mL), THF (10 mL), and water (3 mL), was added LiOH H2O (0.8 g, 0.0154). The reaction mixture was stirred 16 h. Subsequently, the reaction mixture was concentrated under vacuum and neutralized with 1.5 N HCl. The solids were isolated by filtration, washed with water, and dried under vacuum, affording crude DTx-01-32. Recrystallization (80% DCM in hexane) yielded lipid motif DTx-01-32 as an off-white solid (2.3 g, 79.3%). LCMS m/z (M+H)+: 567.2; 1H-NMR (400 MHz, TFA-d): δ 0.87-0.98 (m, 6H), 1.20-1.58 (m, 41H), 1.74-1.92 (m, 8H), 2.18-2.21 (m, 2H), 2.73 (t, J=7.6 Hz, 2H), 3.05 (t, J=7.6 Hz, 2H), 3.60 (t, J=7.8 Hz, 2H).
Scheme I above illustrates the preparation of an oligonucleotide conjugated with an uptake motif at the 3′ terminus of the oligonucleotide, i.e. at the 3′ carbon of the terminal 3′ nucleotide. In summary, 3′-amino CPG beads I-1 (Glen Research, Catalog No. 20-2958) modified with the DMT and Fmoc-protected C7 linker illustrated above were treated with 20% piperidine/DMF to afford Fmoc-deprotected amino C7 CPG beads I-2. An uptake motif (e.g. DTx-01-08) was then coupled to 1-2 using HATU and DIEA in DMF to produce lipid-loaded CPG beads 1-3, which were treated by 3% dichloroacetic acid (DCA) in DCM to remove the DMT protecting group and afford I-4. Oligonucleotide synthesis was accomplished via standard phosphoramidite chemistry and yielded oligonucleotide-bounded CPG beads I-5. At this point, if applicable, beads I-5 containing methyl ester-protected lipid motifs (e.g., DTx-01-07-OMe, DTx-01-09-OMe) were saponified to their respective carboxylic acid using 0.5 M LiOH in 3:1 v/v methanol/water. Subsequent treatment of I-5 with AMA [ammonium hydroxide (28%)/methylamine (40%) (1:1, v/v)] cleaved the DTx-01-08-conjugated oligonucleotide from the beads. The conjugated oligonucleotide was then purified by RP-HPLC and characterized by MALDI-TOF MS using the [M+H] peak.
Scheme II above illustrates the preparation of a sense strand of a double-stranded oligonucleotide conjugated with an uptake motif at each of the 5′ and 3′ termini. In summary, 3′-amino CPG beads 11-1 (Glen Research, Catalog No. 20-2958) modified with the DMT and Fmoc-protected C7 linker illustrated above were treated with 20% piperidine/DMF to afford Fmoc-deprotected amino C7 CPG beads II-2. An uptake motif (e.g. DTx-01-08) was then coupled to 11-2 using HATU and DIEA in DMF to produce the fatty-acid loaded CPG beads II-3, which were subsequently treated with 3% dichloroacetic acid (DCA) in DCM to remove the DMT protecting group and afford 11-4. Oligonucleotide synthesis was performed on 11-4 via standard phosphoramidite chemistry. The final coupling was with a phosphoramidite (Glen Research, Catalog No. 10-1906) that incorporated a monomethoxytrityl (MMTr) protected 6-carbon alkyl amine as shown in structure 11-5. After removal of MMT with 3% dichloroacetic acid (DCA) in DCM, II-6 was coupled to DTx-01-08 using HATU and DIEA in DMF to yield 11-7. Stepwise deprotection with triethylamine in acetonitrile (to remove phosphate protecting groups) and AMA [ammonium hydroxide (28%)/methylamine (40%) (1:1, v/v)] (to remove base protecting groups and cleave the oligonucleotide from the synthesis resin) yielded crude II-8. Purification using RP-HPLC yielded a conjugated oligonucleotide. Purity and identity of 11-8 were confirmed by analytical RP-HPLC and MALDI-TOF MS using the [M+H] peak, respectively.
Scheme III above illustrates the preparation of an oligonucleotide conjugated to an uptake motif at the 5′ terminus, i.e. at the 5′ carbon of the 3′ terminal nucleotide. In summary, oligonucleotide synthesis was performed on CPG beads III-1 (Glen Research, Catalog No. 20-5041-xx) via standard phosphoramidite chemistry. In the last nucleotide coupling of the automated sequence, a nucleotide modified with the MMT-protected C6 linker illustrated above (Glen Research, Catalog No. 10-1906) was used, yielding modified oligonucleotide-bounded CPG beads III-2. After removal of MMT with 3% dichloroacetic acid (DCA) in DCM, 111-2 was coupled to an uptake motif (e.g., DTx-01-08) using HATU and DIEA in DMF to yield III-4. Subsequent treatment with AMA [ammonium hydroxide (28%)/methylamine (40%) (1:1, v/v)] cleaved the DTx-01-08-conjugated modified oligonucleotide from the beads to generate III-5. The oligonucleotide was then purified by RP-HPLC and characterized by MALDI-TOF MS using the [M+H] peak.
For each of the strands synthesized by Schemes I, II, or III and listed above, the corresponding complementary strand was prepared via standard phosphoramidite chemistry, purified by IE-HPLC, and characterized by MALDI-TOF MS using the [M+H] peak. The duplex was formed by mixing equal molar equivalents of the passenger strand (the sense strand) and guide strand (the antisense strand), heating to 90° C. for 5 minutes, and then slowly cooling to room temperature. Duplex formation was confirmed by non-denaturing PAGE or non-denaturing HPLC.
Cell Culture. HEK293 cells were purchased from ATCC and were cultured in DMEM containing 10% Fetal Bovine Serum (FBS), 2 mM L-glutamine, 1× non-essential amino acids, 100 U/mL penicillin and 100 mg/mL streptomycin in a humidified 37° C. incubator with 5% CO2. Human Schwann cells (HSwC), isolated from human spinal nerve and cryopreserved at first passage (P1), were purchased from iXcells Biotechnologies (Cat #10HU-188). HSwC were cultured in Schwann Cell Growth Medium (Cat #MD-0055) in a humidified 37° C. incubator with 5% CO2.
Generation of Stable Human and Mouse PMP22 Cell Lines. 3×106 HEK293 cells were plated onto 10-cm tissue culture treated petri dishes in the media described herein without antibiotics. The day after plating, human (Origene, Cat #RC216500) or mouse (Origene, Cat #MR225485) PMP22 plasmids were transfected into HEK293 cells with Lipofectamine 2000 according to the manufacturer's protocol. Briefly, 20 ug of each plasmid were diluted in 480 μL of DMEM without FBS or antibiotic. Separately, 50 uL of Lipofectamine 2000 was diluted in 450 uL of DMEM without FBS or antibiotic. The plasmid/DMEM and the Lipofectamine 2000/DMEM cocktails were then combined, mixed by titrating up and down and incubated for 20 minutes at room temperature to enable complex formation. The DMEM media containing FBS but lacking antibiotic (9 mL) was then added to the plasmid/Lipofectamine 2000 complexes (1 mL) and then added to cells in the 10-cm dish. The cells were incubated overnight at 37° C. in the incubator. Media was then removed and replaced with DMEM containing FBS and antibiotic. Five days post-transfection, the media was replaced with DMEM containing FBS, antibiotic and 800 ug/mL geneticin to select for cells that stably express either the human or mouse PMP22. The cells were cultured in this media for 30 days with media changes every 3 days. The cells were then expanded and subsequently cryopreserved. Sequencing and qPCR were utilized to confirm integration of the human or mouse PMP22 expression vector.
Reverse Transfection of siRNA. HEK293 cells were trypsinized and diluted to 20,000 cells/well, in 90 uL of antibiotic-free media. Schwann cells were trypsinized and diluted to 10,000 cells/well, in 90 uL of antibiotic-free media. Compounds were diluted in PBS to 100× of the desired final concentration. Separately, Lipofectamine RNAiMax (Life Technologies) was diluted 1:66.7 in media lacking supplements (e.g. FBS, antibiotic etc.). The 100× compound in PBS was then complexed with RNAiMAX by adding 1 part compound in PBS to 9 parts lipofectamine/media. Following incubation for 20 minutes, 10 uL of the compound:RNAiMAX complexes were added to a 96-well collagen coated plate. A volume of 90 ul of the cell dilution was added to each well of the 96-well plate. The plate was then placed in a humidified 37° C. incubator with 5% CO2. After 24 hours, the complexes were removed and replaced with complete media containing antibiotics for each cell line. HEK293 media was replaced with DMEM containing 10% FBS, 2 mM L-glutamine, 1× non-essential amino acids, 100 U/mL penicillin and 100 mg/mL streptomycin. Schwann cell media was replaced with Schwann Cell Growth Medium. RNA was isolated 48 hours following transfection.
Free uptake of conjugated siRNA. HEK293 cells were trypsinized and diluted to 20,000 cells/well, in 100 uL of complete media and allowed to settle overnight in 96 well collagen coated plates. Schwann cells were trypsinized and diluted to 10,000 cells/well, in 100 uL of complete media and allowed to settle for 48 hours in 96 well collagen coated plates. Compounds were diluted in deep well plates in the corresponding basal media for each cell line supplemented with 2% FBS to the desired final concentration of the top dose then serially diluted. After the appropriate amount of time for cells to settle, media was removed from plates by inverting. 100 ul of compound or PBS at proper concentrations was added to each well of the 96 well plate. HEK293 cells were incubated for 48 hours, and Schwann cells were incubated 72 hours in a humidified 37° C. incubator with 5% CO2 before RNA was isolated.
RNA Isolation, Reverse Transcription and Quantitative PCR. RNA was isolated utilizing the RNeasy 96 kit (Qiagen) according to the manufacturer's protocol. RNA was reverse transcribed to cDNA utilizing random primers and the high-capacity cDNA reverse transcription kit (ThermoFisher Scientific) in a SimpliAmp thermal cycler (ThermoFisher Scientific) according to the manufacturer's instructions. Real-time quantitative PCR was performed utilizing gene-specific primers (Thermofisher Scientific; IDTDNA), TaqMan probes (Thermofisher Scientific; IDTDNA) and TaqMan fast universal PCR master mix (Thermofisher scientific) on a StepOnePlus real-time PCR system (Thermofisher Scientific) according to the manufacturer's instructions. For analysis of quantitative PCR, mRNA expression was normalized to the expression of either 18s rRNA, b-actin or HPRT1 mRNA (housekeeping genes) utilizing the relative CT method according to the best practices proposed in Nature Protocols (Schmittgen, T. D. & Livak, K. J. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 3, 1101-1108 (2008)).
Mice. C3-PMP22 (B6.Cg-Tg(PMP22)C3Fbas/J) male mice were originally purchased from the Jackson Laboratory. C3-PMP22 mice express 3 to 4 copies of a wild-type human peripheral myelin protein 22 (PMP22). The C3-PMP22 male mice were used to set up a mouse colony. The transgenic line was maintained hemizygous by breeding C3-PMP22 males with wildtype females (C57BL/6J). All litters were weaned between 21-23 days of age and tail clipped for genotyping. Both hemizygous female and male mice were used for experiments.
Intravenous injection. Mice were weighed the day before the study initiation. On the day of the study, the mice were restrained with an approved device and injected with the treatment of interest (compound or PBS) via the tail vein.
Target Engagement Studies in vivo in wildtype mice and C3-PMP22 mice. 7-84 days following intravenous injection of the compound of interest or control, the mice were euthanized. Sciatic, tibial, sensory, and motor branches of the femoral nerves and/or brachial plexus were dissected and prepared for RNA isolation. The regions of interest were placed in tubes containing beads, flash frozen and stored at −80° C. until RNA isolation. To extract total RNA, Trizol was added to the tubes and RNA isolated using the RNeasy 96 kit via the manufacturer's instructions.
Electrophysiology assessment using Electromyography (EMG). The EMG apparatus (ADInstruments, PowerLab Cat #PL2604/P) was used to measure motor nerve conduction velocity (MNCV). The mice were anesthetized in an isoflurane chamber and transferred to the nose cone on a recirculating water heating pad to maintain their temperature. A rectal probe was used to monitor the temperature. A total of 4 electrodes were used: 2 recording and 2 stimulating electrodes. The two recording electrodes were gently inserted between the 1st and 2nd and 2nd and 3rd toes and taped to the plexiglass surface. One stimulating electrode was inserted under the skin between the shoulders. The second stimulating electrode was inserted into the skin of the ankle. The EMG was set to deliver a stimulus using a 0.1 msec square pulse stimulus every 2 seconds. The stimulation voltage was gradually increased until the maximal M-wave is observed (Mmax). The stimulating electrode was then moved from the ankle to the greater sciatic notch and stimulate once. The stimulation was repeated at the ankle and sciatic notch 2 more times each. At the end of the last measurement, leaving the electrode at the hip, the electrodes from the toes were removed and the leg stretched. A compass was used to measure the distance between the electrode at the hip and the point at the ankle at which stimulation was conducted. The latency between the M-wave in response to stimulation at the ankle vs hip was calculated and averaged across the 3 trials. This value was divided by the distance between the electrodes to calculate the motor conduction velocity. At the end of the measurement all electrodes were removed, and the mouse was placed on a water-recirculating heating pad that is set at 37° C. Once the mouse has fully recovered it was returned to housing rack in animal holding room.
Myelin staining. The nerves of interest were carefully dissected, placed lengthwise on a stick of wood (applicator or matchstick) to prevent the nerve from folding, and immersed in a scintillation vial containing cold 2.5% glutaraldehyde (fixative) overnight at 4° C. The following day the nerves were washed with 0.1M sodium phosphate buffer and immersed in 2% osmium for approximately 1 hour (osmium penetrates tissue from all sides at roughly 0.5 mm/hr, so a mouse nerve with a diameter of 1 mm should osmicate for 1 hour). After rinsing in water, the nerves were dehydrated and embedded in resin blocks. Once embedded in resin blocks the nerves were cut with glass knifes using a microtome in 0.15 um sections. The sections were subsequently stained with 2% paraphenylenediamine (PPD) for 20 minutes at room temperature, rinsed, dried and coverslip mounted for microscopic examination.
Beam Walking. Coordination and balance were evaluated through the beam walking assay by two experimenters that were blinded to experimental conditions. Mice were trained over two-three consecutive days to cross a 100 cm-long painted wood round beam with a 25 mm diameter to reach a platform with a darkened escape box. The beam was place 30 cm over a padded surface. Training trials ended when the mouse reached the escape platform or when the mouse fell off the beam. The latency to cross the beam and the number of times the hind paws slipped during placement were tabulated for each training run. Each training run was repeated three times per day with a minimum of 5 minutes between runs. Training was considered complete when all mice crossed the beam consistently without pausing. On the subsequent testing day, mice underwent three trials in which they crossed the 25 mm-diameter beam, with a minimum of 5 minutes between runs. Then mice underwent an additional three trials in which they crossed a 10 mm-diameter beam. Latency to cross the beam and the number of foot slips or falls were tabulated for each trial. Data from the second and third trials on each beam were averaged. Trials in which the mouse paused while crossing or fell off the beam were excluded from analysis.
Hindlimb clasping. In order to evaluate general neuromuscular dysfunction, incidence of hindlimb clasping was observed. A blinded observer took a photo of hindlimb behavior while suspending the mice briefly from their tails. From these images, hindlimb behavior was scored as 0-normal splaying of the hindlimbs and toes of the paw spread wide, 1-clasping of one foot or hindlimb, or 2-clasping of both feet of hindlimb. The angle of hindlimb spread was also calculated from the images using ImageJ2 (NIH, Rueden et al, 2017) to measure the angle between the hind paws by drawing a vector from each paw to the anus.
Grip strength. Grip strength is a measure of muscular strength, or the maximum force/tension generated by one's forearm muscles. It can be measured using a digital force meter equipped with precision force gauges to retain the peak force applied on a digital display and with a grid or wire system that allows mouse grip by either or both paws. Each mouse was lifted by the tail to the height where the front paws are at the same height as the bar/grid. The mouse was then moved horizontally towards the bar/grid until it was within reach. After visually checking that the grip was good, i.e. a symmetric, tight grip with both paws and exerting a detectable resistance against the investigator's pull, the mouse was gently pulled away until its grasp is broken. The pulling was at a constant speed and sufficiently slow to permit the mouse to build up a resistance against it. The transducer saved the value at this point. Measurements were discarded if the animal used only one paw or also used its hind paws, turned backwards during the pull, or released the bar without resistance. The test was repeated three times and the values averaged.
Numerous siRNAs targeting the human PMP22 mRNA were designed and synthesized. The sense and antisense strands of the compounds ere prepared with sugar moiety, terminal, and internucleotide linkage modifications to increase hybridization affinity, minimize degradation by nucleases, and enhance loading into RISC. The siRNAs are shown in Table 3.
In Table 3, “Start” and “End” correspond to the 5′ and 3′ nucleotide positions of the nucleotide sequence of the human PMP22 mRNA (NCBI Reference Sequence NM_000304.4, deposited with GenBank on Nov. 22, 2018; SEQ ID NO: 1170) to which the nucleotides of the antisense strand are complementary. Each row represents a sense and antisense strand pair of an siRNA. If present, an siRNA ID in the “Parent siRNA ID” column indicates an siRNA related by nucleotide sequence.
Modified sugar moieties are indicated by a subscript notation following the nucleotide, and modified internucleotide linkages are indicated by a superscript notation. A nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; and a nucleotide followed by the subscript “D” is a beta-D-deoxyribonucleotide. A superscript “S” is a phosphorothioate internucleotide linkage; all other internucleotide linkages are phosphodiester internucleotide linkages. For example, “UFSCM” is a 2′-flourouridine linked to a 2′-O-methylcytidine by a phosphorothioate internucleotide linkage. “GMUF” is a 2-O-methylguanosine linked to a 2′-fluorouridine by a phosphodiester internucleotide linkage. A hydroxyl group is at the 5′ carbon of the 5′ terminal nucleotide is indicated by “5′-OH”; a phosphate group at the 5′ carbon of the 5′ terminal nucleotide is indicated by “5′-PO4”; and a hydroxyl group at the 3′ carbon of the 3′ terminal nucleotide is indicated by “OH-3′.”
FCMCFUMGFU
MAFGMCFAMA
MUFGMCFUMG
FCMAFGMGFA
FAMGFUMAFST
MGFGMAFGMS
D
STD-OH-3′
FGMGFAMUFC
MCFCMAFCMGF
MGFUMGFGMG
FCMAFAMUFST
D
STD-OH-3′
STD-OH-3′
FGMAFUMCFU
MCFCMAFGMA
MCFUMGFGMC
FGMAFUMCFA
FAMGFAMAFST
MGFUMUFGMS
D
STD-OH-3′
FCMUFUMCFCM
MCFUMGFAMG
FGMAFAMGFA
MGFGMUFGMS
STD-OH-3′
FUMGFUMCFC
MUFGMGFUMG
MAFCMCFAMCF
FGMAFCMAFU
MUFUMCFCMST
STD-OH-3′
D
STD-OH-3′
FUMCFAMUFC
MUFGMAFUMG
MAFUMCFAMC
FAMUFGMAFG
FCMAFAMAFST
MAFAMAFCMST
D
STD-OH-3′
D
STD-OH-3′
FUMCFAMUFC
MUFGMAFAMG
MUFUMCFAMG
FAMUFGMAFU
FCMAFUMUFST
MCFGMAFCMST
D
STD-OH-3′
D
STD-OH-3′
FUMGFUMUFC
MAFGMAFAMG
MUFUMCFUMG
FAMAFCMAFG
MGFAMAFCMST
D
STD-OH-3′
D
STD-OH-3′
FUMGFGMAFA
MAFAMGFAMU
MUFCMUFUMC
FUMCFCMAFG
FCMAFAMAFST
MUFGMAFUM
D
STD-OH-3′
FUMCFUMUFC
MUFUMUFGMG
MCFAMAFAMU
FAMAFGMAFU
MUFCMCFAMST
D
STD-OH-3′
D
STD-OH-3′
FCMUFCMAFA
MCFGMAFGMU
MCFUMCFGMG
FUMGFAMGFA
FAMUFUMAFST
MUFGMCFCMST
D
STD-OH-3′
D
STD-OH-3′
FUMAFCMGFG
MGFAMAFAMC
MUFUMUFCMG
FCMGFUMAFG
MGFAMGFUMS
D
STD-OH-3′
FUMCFUMCFU
MUFGMCFCMA
MGFGMCFAMG
FGMAFGMAFU
FAMAFCMUFST
MCFAMGFUMST
D
STD-OH-3′
D
STD-OH-3′
FCMUFCMUFG
MCFUMGFCMCF
MGFCMAFGMA
FAMCFUMGFST
D
STD-OH-3′
STD-OH-3′
FUMCFUMGFG
MUFCMUFGMC
MCFAMGFAMA
FCMAFGMAFG
MAFUMCFAMST
D
STD-OH-3′
D
STD-OH-3′
FCMUFGMGFC
MUFUMCFUMG
MAFGMAFAMC
FCMCFAMGFA
FUMGFUMAFST
MGFAMUFCMST
D
STD-OH-3′
D
STD-OH-3′
FGMUFCMCFA
MGFGMCFCMU
MGFGMCFCMA
FGMGFAMCFA
FCMCFAMUFST
MGFAMCFUMST
D
STD-OH-3′
D
STD-OH-3′
FCMCFAMGFG
MGFUMGFGMC
MCFCMAFCMCF
FCMUFGMGFA
MCFAMGFAMST
STD-OH-3′
D
STD-OH-3′
FCMAFGMGFC
MGFGMUFGMG
MCFAMCFCMAF
FCMCFUMGFG
MAFCMAFGMST
STD-OH-3′
D
STD-OH-3′
FCMAFCMCFAM
MAFUMCFAMU
MCFCMUFGMST
STD-OH-3′
D
STD-OH-3′
FUMUFCMCFU
MGFAMAFCMA
MGFUMUFCMU
FGMGFAMAFC
FUMCFUMGEST
MAFGMAFGMS
D
STD-OH-3′
FUMCFCMUFG
MAFGMAFAMC
MUFUMCFUMU
FAMGFGMAFA
FCMUFGMCFST
MCFAMGFAMST
D
STD-OH-3′
D
STD-OH-3′
FGMGFUMCFU
MGFCMAFCMA
MGFUMGFCMG
FGMAFCMCFA
FUMGFAMUFST
MGFCMAFAMST
D
STD-OH-3′
D
STD-OH-3′
FUMGFUMGFC
MUFCMAFCMG
MGFUMGFAMU
FCMAFCMAFG
FGMAFGMUFST
MAFCMCFAMST
D
STD-OH-3′
D
STD-OH-3′
FUMUFGMCFU
MAFCMUFCMA
MGFAMGFUMA
FGMCFAMAFC
FUMCFAMUFST
MAFGMGFAMS
D
STD-OH-3′
FGMUFCMAFG
MUFUMGFGMC
MCFCMAFAMU
FUMGFAMCFG
FGMGFAMUFST
MAFUMCFGMST
D
STD-OH-3′
D
STD-OH-3′
FCMUFGMUFA
MGFUMGFCMU
MGFCMAFCMCF
FAMCFAMGFU
MUFCMUFGMST
STD-OH-3′
D
STD-OH-3′
FUMCFCMUFCM
MUFCMCFUMG
FAMGFGMAFA
MGFAMGFGyS
STD-OH-3′
FAMUFCMAFC
MUFUMUFGMG
MCFAMAFAMC
FUMGFAMUFG
FGMAFAMUFST
MAFUMGFAMS
D
STD-OH-3′
FAMUFCMCFU
MCFGMAFCMA
MGFUMCFGMA
MAFUMGFGMS
D
STD-OH-3′
FAMUFUMCFU
MAFGMAFCMA
MGFUMCFUMC
FUMGFUMUFST
MCFUMGFAMST
D
STD-OH-3′
D
STD-OH-3′
FGMUFUMCFC
MAFAMCFAMG
MUFGMUFUMC
FGMAFAMCFA
FUMUFCMUFST
MGFAMGFAMS
D
STD-OH-3′
FGMGFAMAFU
MGFAMAFGMA
MCFUMUFCMCF
FUMUFCMCFA
MGFUMGFAMS
STD-OH-3′
FAMUFCMUFU
MUFUMGFGMA
MCFCMAFAMA
FAMGFAMUFU
FUMUFCMUFST
MCFCMAFGMST
D
STD-OH-3′
D
STD-OH-3′
FAMGFCMGFG
MGFAMCFAMC
MUFGMUFCMA
FCMGFCMUFG
FUMCFUMAFST
MAFGMAFAMS
D
STD-OH-3′
FGMCFGMGFU
MUFGMAFCMA
MGFUMCFAMU
FCMCFGMCFUM
FCMUFAMUFST
D
STD-OH-3′
STD-OH-3′
FUMCFCMUFG
MGFCMAFAMC
MUFUMGFCMU
FAMGFGMAFG
FGMAFGMUFST
MGFAMGFCMST
D
STD-OH-3′
D
STD-OH-3′
FCMUFGMUFU
MCFAMGFCMA
MGFCMUFGMA
FAMCFAMGFG
FGMUFAMUFST
MAFGMGFAMS
D
STD-OH-3′
FUMGFUMUFG
MUFCMAFGMC
MCFUMGFAMG
FAMAFCMAFG
FUMAFUMCEST
MGFAMGFGMS
D
STD-OH-3′
MCFUMCFAMG
MUFGMAFGMU
FCMAFAMCFA
FAMUFCMAFST
MGFGMAFGMS
D
STD-OH-3′
FUMGFCMUFG
MUFAMCFUMC
MAFGMUFAMU
FAMGFCMAFA
MCFAMGFGMST
D
STD-OH-3′
D
STD-OH-3′
MAFUMAFCMU
MGFUMAFUMC
FCMAFGMCFA
FAMUFCMGEST
MAFCMAFGMST
D
STD-OH-3′
D
STD-OH-3′
FCMUFGMAFG
MGFAMUFAMC
MUFAMUFCMA
FUMCFAMGFC
FUMCFGMUFST
MAFAMCFAMST
D
STD-OH-3′
D
STD-OH-3′
FUMGFAMGFU
MUFGMAFUMA
MAFUMCFAMU
FCMUFCMAFG
FCMGFUMCFST
MCFAMAFCMST
D
STD-OH-3′
D
STD-OH-3′
FGMAFGMUFA
MAFUMGFAMU
MUFCMAFUMC
FAMCFUMCFA
FGMUFCMCFST
MGFCMAFAMST
D
STD-OH-3′
D
STD-OH-3′
FAMUFCMAFU
MGFAMCFGMA
MCFGMUFCMCF
FUMGFAMUFA
MCFUMCFAMST
STD-OH-3′
D
STD-OH-3′
FAMUFCMGFU
MGFAMGFGMA
MCFCMUFCMCF
FCMGFAMUFG
MAFUMAFCMST
STD-OH-3′
D
STD-OH-3′
FGMUFGMCFU
MCFAMCFCMAF
MGFGMUFGMC
FUMGFCMUFST
D
STD-OH-3′
STD-OH-3′
FCMUFGMGFU
MCFAMGFCMA
MGFCMUFGMC
FCMCFAMGFCM
FUMGFUMUFST
D
STD-OH-3′
STD-OH-3′
MAFAMCFAMG
MUFGMUFUMC
FCMAFGMCFA
FGMUFCMUFST
MCFCMAFGMST
D
STD-OH-3′
D
STD-OH-3′
FGMCFUMGFU
MAFCMGFAMA
MUFCMGFUMC
FCMAFGMCFA
FUMCFCMAFST
MGFCMAFCMST
D
STD-OH-3′
D
STD-OH-3′
FUMUFCMGFU
MGFGMAFGMA
MCFUMCFCMAF
FCMGFAMAFC
MAFGMCFAMST
STD-OH-3′
D
STD-OH-3′
FCMUFCMCFAM
MAFUMCFGMU
FGMGFAMGFA
MCFGMAFAMST
STD-OH-3′
D
STD-OH-3′
FGMAFUMCFG
MCFUMGFAMC
MUFCMAFGMC
FGMAFUMCFG
FCMAFAMUFST
MUFGMGFAM
D
STD-OH-3′
FUMGFGMAFU
MCFAMCFGMA
MCFGMUFGMG
FUMCFCMAFU
FGMCFAMAFST
MUFGMGFCMST
D
STD-OH-3′
D
STD-OH-3′
FCMAFAMUFG
MUFGMUFCMC
MGFAMCFAMC
FAMUFUMGFC
FGMCFAMAFST
MCFCMAFCMST
D
STD-OH-3′
D
STD-OH-3′
FGMAFCMAFC
MUFUMGFCMG
MGFCMAFAMC
FUMGFUMCFC
FUMGFAMUFST
MAFUMUFGMS
D
STD-OH-3′
FCMGFCMAFA
MUFCMAFGMU
MCFUMGFAMU
FUMGFCMGFU
FCMUFCMUFST
MGFUMCFCMST
D
STD-OH-3′
D
STD-OH-3′
FUMCFUMUFC
MUFGMAFGMG
MCFUMCFAMG
FAMAFGMAFG
FGMAFAMAFST
MGFUMGFCMST
D
STD-OH-3′
D
STD-OH-3′
FGMUFCMCFA
MGFUMGFGMU
MCFCMAFCMUF
FGMGFAMCFA
MUFUMUFCMST
STD-OH-3′
D
STD-OH-3′
FCMUFGMUFU
MGFAMGFAMA
MUFCMUFCMA
FAMCFAMGFU
FUMCFAMUFST
MGFGMUFGMS
D
STD-OH-3′
FCMUFCMAFU
MGFAMUFGMA
MCFAMUFCMA
FUMGFAMGFA
FCMCFAMAFST
MAFAMCFAMST
D
STD-OH-3′
D
STD-OH-3′
FUMCFAMCFCM
MGFUMUFUMG
FGMUFGMAFU
MGFAMUFGMS
STD-OH-3′
FUMGFGMCFU
MCFUMGFCMA
MGFCMAFGMU
FGMCFCMAFU
FCMUFGMUFST
MUFCMGFUMST
D
STD-OH-3′
D
STD-OH-3′
FAMUFGMAFU
MCFAMGFGMA
MCFCMUFGMU
FUMCFAMUFG
MGFUMGFGMS
D
STD-OH-3′
FCMUFGMUFC
MGFAMUFCMG
MGFAMUFCMA
FAMCFAMGFG
FUMCFUMUFST
MAFUMCFAMST
D
STD-OH-3′
D
STD-OH-3′
FAMUFCMAFU
MGFAMAFGMA
MCFUMUFCMA
FUMGFAMUFC
FGMCFAMUFST
MGFAMCFAMST
D
STD-OH-3′
D
STD-OH-3′
FUMUFCMUFU
MGFCMAFGMA
MCFUMGFCMCF
FAMGFAMAFC
MAFGMGFAMS
STD-OH-3′
FUMUFCMUFG
MUFUMGFGMC
MCFCMAFAMCF
FAMGFAMAFG
MAFAMCFAMST
STD-OH-3′
D
STD-OH-3′
FAMCFUMCFU
MGFUMGFAMA
MUFCMAFCMCF
FGMAFGMUFU
MGFGMCFAMST
STD-OH-3′
D
STD-OH-3′
FUMUFCMAFC
MGFAMGFGMG
MCFCMUFCMAR
FUMGFAMAFG
MAFGMUFUM
STD-OH-3′
FGMGFUMUFU
MUFGMUFAMA
MUFAMCFAMU
FAMAFCMCFU
FCMAFCMUFST
MGFCMCFCMST
D
STD-OH-3′
D
STD-OH-3′
FUMAFCMAFU
MAFGMUFGMA
MCFAMCFUMG
FUMGFUMAFA
FGMAFAMUFST
MAFAMCFCMST
D
STD-OH-3′
D
STD-OH-3′
FAMUFCMAFC
MUFCMCFAMG
MUFGMGFAMA
FUMGFAMUFG
FUMCFUMUFST
MUFAMAFAMS
D
STD-OH-3′
FGMAFAMUFC
MGFGMAFAMG
MUFUMCFCMA
FAMUFUMCFC
FAMAFUMUFST
MAFGMUFGMS
D
STD-OH-3′
FUMCFCMAFA
MGFAMAFUMU
MAFUMUFCMU
FUMGFGMAFA
FUMGFCMUFST
MGFAMUFUMS
D
STD-OH-3′
FUMCFUMUFG
MCFCMAFGMCF
MCFUMGFGMU
D
STD-OH-3′
STD-OH-3′
FUMCFUMGFU
MAFCMGFCMA
MGFCMGFUMG
FCMAFGMAFC
FAMUFGMAFST
MCFAMGFCMST
D
STD-OH-3′
D
STD-OH-3′
FAMGFUMGFC
MCFGMCFAMG
MUFGMCFGMG
FCMCFAMUFST
D
STD-OH-3′
STD-OH-3′
MGFGMCFCMG
MGFGMCFCMA
FCMAFGMCFA
FUMCFUMAFST
MCFUMCFAMST
D
STD-OH-3′
D
STD-OH-3′
MGFUMGFUMA
MAFCMAFCMG
FGMAFUMGFG
FGMUFGMAFST
MCFCMGFCMST
D
STD-OH-3′
D
STD-OH-3′
MGFGMGFUMG
MAFCMCFCMGF
FCMCFUMCFAM
STD-OH-3′
STD-OH-3′
FAMCFCMCFGM
MAFCMUFCMCF
STD-OH-3′
STD-OH-3′
MAFUMGFCMC
MGFCMAFUMC
FAMCFUMCFCM
FUMCFAMAFST
D
STD-OH-3′
STD-OH-3′
FUMCFUMCFA
MGFAMGFUMU
MAFCMUFCMG
FGMAFGMAFU
FGMAFUMUFST
MGFCMCFAMST
D
STD-OH-3′
D
STD-OH-3′
FUMCFGMGFA
MGFUMAFAMU
MUFUMAFCMU
FCMCFUMAFST
MUFUMGFAMS
D
STD-OH-3′
FUMAFCMUFC
MGFUMAFGMG
MCFUMAFCMG
FAMGFUMAFA
FGMUFUMUFST
MUFCMCFGMST
D
STD-OH-3′
D
STD-OH-3′
FGMGFUMUFU
MGFGMCFGMA
MCFGMCFCMUF
FAMAFCMCFG
MUFAMGFGM
STD-OH-3′
FUMUFCMGFC
MGFUMAFGMG
MCFUMAFCMA
FUMCFCMUFST
MCFCMGFUMST
D
STD-OH-3′
D
STD-OH-3′
FGMCFCMUFG
MCFAMCFCMCF
MGFGMUFGMG
FCMCFUMUFST
D
STD-OH-3′
STD-OH-3′
FCMUFCMAFG
MAFCMCFGMCF
MCFGMGFUMG
FUMCFAMUFST
D
STD-OH-3′
STD-OH-3′
FGMUFCMAFU
MAFUMAFGMA
MCFUMAFUMG
FUMGFAMCFA
FUMGFAMUFST
MCFCMGFCMST
D
STD-OH-3′
D
STD-OH-3′
FAMUFCMUFA
MCFAMCFAMU
MUFGMUFGMA
FAMGFAMUFG
FUMCFUMUFST
MAFCMAFCMST
D
STD-OH-3′
D
STD-OH-3′
FUMGFAMUFC
MGFCMAFAMG
MUFUMGFCMG
FAMUFCMAFC
FGMAFAMAFST
MAFUMAFGMS
D
STD-OH-3′
FGMCFGMGFA
MCFGMUFUMU
MAFAMCFGMC
FCMCFGMCFAM
FGMAFAMUFST
D
STD-OH-3′
STD-OH-3′
FUMCFUMGFA
MAFCMGFCMU
MGFCMGFUMA
FCMAFGMAFG
FCMAFUMAFST
MCFCMUFCMST
D
STD-OH-3′
D
STD-OH-3′
FAMGFGMGFA
MGFUMUFUMU
MAFAMAFCMA
FCMCFCMUFUM
FGMAFAMAFST
D
STD-OH-3′
STD-OH-3′
FAMAFUMCFC
MUFUMUFGMG
MCFAMAFAMC
FGMAFUMUFU
FUMCFAMAFST
MUFGMGFGMS
D
STD-OH-3′
FAMAFAMCFU
MUFUMUFGMA
MCFAMAFAMC
FGMUFUMUFG
FCMAFAMAFST
MGFGMAFUMS
D
STD-OH-3′
FUMGFAMUFU
MCFUMUFCMA
MGFAMAFGMA
FAMUFCMAFA
FUMGFUMAFST
MCFAMGFCMST
D
STD-OH-3′
D
STD-OH-3′
FGMAFUMUFG
MUFCMUFUMC
MAFAMGFAMU
FAMAFUMCFA
FGMUFAMUFST
MAFCMAFGMST
D
STD-OH-3′
D
STD-OH-3′
FUMUFUMAFU
MUFUMUFUMA
MAFAMAFAMC
FUMAFAMAFC
FCMUFAMUFST
MCFGMGFAMST
D
STD-OH-3′
D
STD-OH-3′
FUMAFAMAFA
MUFAMGFGMU
MCFCMUFAMU
FUMUFUMAFU
FUMUFAMUFST
MAFAMAFCMST
D
STD-OH-3′
D
STD-OH-3′
FGMUFAMUFU
MAFAMAFCMA
MGFUMUFUMG
FAMUFAMCFU
FCMUFUMUFST
MAFUMGFUMS
D
STD-OH-3′
FAMUFCMAFG
MGFAMGFGMC
MCFCMUFCMGF
FUMGFAMUFG
MGFUMCFAMST
STD-OH-3′
D
STD-OH-3′
FAMAFAMGFA
MUFAMCFUMU
MAFGMUFAMG
FCMUFUMUFA
FCMUFAMAFST
MAFGMGFCMST
D
STD-OH-3′
D
STD-OH-3′
FAMGFCMUFA
MUFCMCFUMU
MAFGMGFAMA
FAMGFCMUFA
FCMUFUMUFST
MCFUMUFCMST
D
STD-OH-3′
D
STD-OH-3′
FAMCFUMUFU
MAFUMGFUMA
MAFCMAFUMC
FAMAFGMUFU
FCMUFAMAFST
MCFCMUFUMST
D
STD-OH-3′
D
STD-OH-3′
FUMCFCMUFA
MCFUMGFUMU
MAFCMAFGMU
FAMGFGMAFU
FAMUFAMAFST
MGFUMAFAMS
D
STD-OH-3′
FCMCFUMAFA
MAFCMUFGMU
MCFAMGFUMA
FUMAFGMGFA
FUMAFAMUFST
MUFGMUFAMS
D
STD-OH-3′
FCMAFGMAFA
MUFUMAFUMU
MAFUMAFAMG
FUMCFUMGFG
FAMUFAMAFST
MGFUMAFAMS
D
STD-OH-3′
FCMCFCMUFUM
MAFUMGFAMA
FAMGFGMGFA
MAFGMGFGS
STD-OH-3′
FGMCFAMUFC
MGFUMUFGMG
MCFAMAFCMA
FAMUFGMCFA
FGMAFAMAFST
MCFUMGFGMST
D
STD-OH-3′
D
STD-OH-3′
FUMGFUMGFU
MCFUMUFCMA
MGFAMAFGMC
FCMAFCMAFG
FUMUFUMAFST
MAFGMGFUMS
D
STD-OH-3′
FAMAFCMUFG
MUFCMUFAMC
MUFAMGFAMU
FAMGFUMUFG
FGMUFAMUFST
MGFUMGFGMS
D
STD-OH-3′
FGMCFUMAFA
MAFGMUFCMU
MGFAMCFUMC
FUMAFGMCFA
FCMAFGMAFST
MUFCMAFGMST
D
STD-OH-3′
D
STD-OH-3′
FUMGFCMAFU
MGFAMAFAMA
MUFUMUFCMU
FUMGFCMAFA
FGMAFUMUFST
MAFGMCFAMST
D
STD-OH-3′
D
STD-OH-3′
FAMCFUMGFU
MCFCMAFCMAF
MGFUMGFGMA
FCMUFAMAFST
D
STD-OH-3′
STD-OH-3′
FGMUFGMUFG
MAFGMUFCMC
MGFAMCFUMA
FAMCFAMCFA
FAMGFAMUFST
MGFUMUFGMS
D
STD-OH-3′
FGMGFAMCFU
MUFCMUFUMA
MAFAMGFAMU
FGMUFCMCFA
FGMCFAMUFST
MCFAMCFAMST
D
STD-OH-3′
D
STD-OH-3′
FGMCFAMCFCM
MUFGMGFUMG
FGMUFGMCFU
MCFCMCFUMST
STD-OH-3′
D
STD-OH-3′
FCMCFAMCFCM
MCFCMCFUMGE
STD-OH-3′
STD-OH-3′
FGMGFUMUFG
MCFUMUFCMCF
MGFAMAFGMC
FUMGFCMAFST
D
STD-OH-3′
STD-OH-3′
FAMAFGMCFU
MCFUMGFCMA
MGFCMAFGMG
FGMCFUMUFC
FCMUFUMAFST
MCFAMAFCMST
D
STD-OH-3′
D
STD-OH-3′
FGMCFUMGFC
MGFCMCFUMG
MAFGMGFCMU
FCMAFGMCFU
FUMAFGMUFST
MUFCMCFAMST
D
STD-OH-3′
D
STD-OH-3′
FCMAFGMGFC
MCFUMAFAMG
MUFUMAFGMU
FCMCFUMGFCM
FCMUFGMUFST
D
STD-OH-3′
STD-OH-3′
FUMCFUMGFA
MGFCMAFGMU
MCFUMGFCMCF
FCMAFGMAFG
MAFCMCFCMST
STD-OH-3′
D
STD-OH-3′
FUMCFUMUFG
MAFAMGFGMC
MCFCMUFUMA
FAMAFGMAFC
FAMCFAMUFST
MCFCMUFCMST
D
STD-OH-3′
D
STD-OH-3′
FCMCFUMUFA
MAFUMGFUMU
MAFCMAFUMC
FAMAFGMGFC
MAFAMGFAMS
D
STD-OH-3′
FAMAFCMAFU
MAFGMGFGMA
MCFCMCFUMUF
FUMGFUMUFA
MAFGMGFCMST
STD-OH-3′
D
STD-OH-3′
FCMAFUMCFCM
MCFAMAFGMG
FGMAFUMGFU
MUFAMAFGMS
STD-OH-3′
FGMCFAMUFU
MGFCMCFAMA
MUFGMGFCMU
FAMUFGMCFA
FGMCFAMAFST
MAFGMGFGMS
D
STD-OH-3′
FCMAFUMUFU
MAFGMCFCMA
MGFGMCFUMG
FAMAFUMGFC
FCMAFAMAFST
MAFAMGFGMS
D
STD-OH-3′
FUMUFGMGFC
MUFGMCFAMG
MUFGMCFAMA
FAMGFAMAFST
MUFGMCFAMST
D
STD-OH-3′
D
STD-OH-3′
FUMGFGMCFU
MUFUMGFCMA
MGFCMAFAMA
FGMCFCMAFA
FGMAFAMAFST
MAFUMGFCMST
D
STD-OH-3′
D
STD-OH-3′
FGMGFCMUFG
MUFUMUFGMC
MCFAMAFAMG
FAMGFCMCFA
FAMAFAMUFST
MAFAMUFGM
D
STD-OH-3′
FAMAFAMGFA
MGFAMUFUMU
MAFAMUFCMU
FCMUFUMUFG
FGMCFUMUFST
MCFAMGFCMST
D
STD-OH-3′
D
STD-OH-3′
FAMAFAMUFC
MAFGMCFAMG
MUFGMCFUMU
FAMUFUMUFC
FGMGFAMAFST
MUFUMUFGMS
D
STD-OH-3′
FUMCFUMGFC
MCFCMAFAMG
MUFUMGFGMA
FCMAFGMAFU
FAMGFAMAFST
MUFUMCFUMST
D
STD-OH-3′
D
STD-OH-3′
MCFCMUFUMCE
MAFAMGFGMG
D
STD-OH-3′
STD-OH-3′
FUMGFGMCFC
MGFCMCFCMGF
MGFGMGFCMA
FGMAFAMAFST
D
STD-OH-3′
STD-OH-3′
FUMCFCMGFCM
MCFUMCFAMG
FCMGFGMAFG
MUFUMUFCMST
STD-OH-3′
D
STD-OH-3′
FGMCFUMGFA
MCFUMGFCMU
MGFCMAFGMA
FCMAFGMCFG
FAMCFUMUFST
MGFAMGFUM
D
STD-OH-3′
FAMCFUMUFG
MGFCMGFGMC
MCFCMGFCMCF
FAMAFGMUFU
MCFUMGFCMST
STD-OH-3′
D
STD-OH-3′
MGFGMCFGMG
MCFGMCFCMAF
FCMAFAMGFU
MUFCMUFGMST
STD-OH-3′
D
STD-OH-3′
FUMGFUMUFC
MAFCMAFGMG
MCFUMGFUMU
FAMAFCMAFG
FCMUFUMCFST
MAFGMCFCMST
D
STD-OH-3′
D
STD-OH-3′
MUFGMUFUMA
MAFAMCFAMC
FUMAFAMAFU
FUMUFUMUFST
MAFGMGFUMS
D
STD-OH-3′
FAMAFAMUFA
MGFAMUFUMU
MAFAMUFCMU
FAMUFUMUFA
FCMAFAMAFST
MUFUMGFUMS
D
STD-OH-3′
FUMGFUMUFG
MGFAMUFUMC
MAFAMUFCMU
FAMAFCMAFC
FUMAFAMAFST
MGFAMGFGMS
D
STD-OH-3′
FAMAFCMUFG
MCFAMCFAMCF
MUFGMUFGMG
FAMCFUMAFST
D
STD-OH-3′
STD-OH-3′
FUMAFUMUFU
MUFUMAFUMA
MAFUMAFAMC
FAMAFUMAFG
FAMCFUMUFST
MGFUMUFUMS
D
STD-OH-3′
FGMAFAMGFG
MGFUMUFUMU
MGFAMAFAMA
FCMCFCMUFUM
FCMAFGMAFSA
M
SAF-OH-3′
FAMAFAMAFU
MUFUMUFGMG
MCFCMCFAMAF
FGMAFUMUFU
MUFGMGFGMC
FUMSCMSGM-
FGMUFUMGFA
MCFUMUFCMA
MUFUMGFAMA
FAMUFCMAFA
FGMAFUMGFSU
MCFAMGFCMA
M
SAF-OH-3′
FAMSCMSCM-
FUMUFAMAFA
MUFAMCFUMU
MGFAMAFGMU
FAMGFCMUFSA
MAFGMGFCMU
SAF-OH-3′
FGMAFAMGFG
MGFUMUFUMU
MGFAMAFAMA
FCMCFCMUFUM
FCMAFGMAFSA
M
SAF-OH-3′
FAMAFAMAFU
MUFUMUFGMG
MCFCMCFAMAF
FGMAFUMUFU
MUFGMGFGMC
FUMSCMSGM-
FGMUFUMGFA
MCFUMUFCMA
MUFUMGFAMA
FAMUFCMAFA
MCFAMGFCMA
MsAF-OH-3′
FAMSCMSCM-
FUMUFAMAFA
MUFAMCFUMU
MGFAMAFGMU
FAMGFCMUFSA
MAFGMGFCMU
M
SAF-OH-3′
FCMSAMSAM-
FCMAFUMCFCM
MCFUMGFUMU
FAMGFGMAFU
MGFUMAFAMA
M
SAF-OH-3′
FCMCFCMAFGM
MUFUMAFUMU
FUMCFUMGFG
MGFUMAFAMA
MsAF-OH-3′
FAMSCMSAM-
FCMCFAMAFCM
MUFCMUFAMC
FAMGFUMUFG
MGFUMGFGMC
M
SUF-OH-3′
FUMUFUMGFC
MGFAMAFAMA
MAFUMUFUMU
FUMGFCMAFA
FCMUFGMAFSU
MAFGMCFAMA
M
SUF-OH-3′
FCMUFGMUFG
MAFGMUFCMC
MUFGMGFAMC
FAMCFAMCFA
FUMAFAMGESA
MGFUMUFGMG
M
SUF-OH-3′
FUMSAMSUM-
FCMCFUMCFCM
MGFCMAFAMC
FAMGFGMAFG
MGFAMGFCMA
M
SUF-OH-3′
FUMSUMSCM-
FUMCFCMUFG
MCFAMGFCMA
MUFUMGFCMU
FAMCFAMGFG
FGMAFGMUFSA
MAFGMGFAMG
M
SUF-OH-3′
FCMSAMSUM-
FCMCFUMGFU
MUFCMAFGMC
MUFGMCFUMG
FAMAFCMAFG
FAMGFUMAFSU
MGFAMGFGMA
M
SCF-OH-3′
FGMSCMSAM-
MCFUMCFAMG
MGFCMUFGMA
FCMAFAMCFA
FGMUFAMUFSC
MGFGMAFGMG
M
SAF-OH-3′
FAMSGMSCM-
FGMUFUMGFC
MUFAMCFUMC
MUFGMAFGMU
FAMGFCMAFA
FAMUFCMAFSU
MCFAMGFGMA
M
SCF-OH-3′
FUMUFGMCFU
MAFUMAFCMU
MGFAMGFUMA
FCMAFGMCFA
FUMCFAMUFSC
MAFCMAFGMG
M
SGF-OH-3′
FAMSGMSGM-
FGMCFUMGFA
MUFGMAFUMA
MGFUMAFUMC
FCMUFCMAFG
FAMUFCMGFSU
MCFAMAFCMA
M
SCF-OH-3′
MAFUMGFAMU
MUFAMUFCMA
FAMCFUMCFA
FUMCFGMUFSC
MGFCMAFAMC
M
SCF-OH-3′
FAMSGMSGM-
FUMCFAMUFC
MGFAMGFGMA
MGFUMCFCMU
FCMGFAMUFG
FCMCFAMCFSG
MAFUMAFCMU
M
SUF-OH-3′
FCMGFGMUFG
MCFAMCFCMAF
MCFUMGFGMU
FGMCFUMGFSC
M
SUF-OH-3′
MAFAMCFAMG
MGFCMUFGMU
FUMCFGMUFSC
MCFCMAFGMCF
M
SUF-OH-3′
FUMGFGMAFC
MUFUMGFCMG
MAFCMGFCMA
FUMGFUMCFC
FAMCFUMGFA
MAFUMUFGMC
MUF-OH-3′
FCMSCMSAM-
FCMAFCMGFCM
MUFCMAFGMU
FUMGFCMGFU
MGFUMCFCMA
M
SUF-OH-3′
FAMUFGMUFC
MGFUMGFGMU
MCFAMCFCMAR
FGMGFAMCFA
MUFUMUFCMC
FUMSGMSAM-
FCMAFCMUFG
MGFAMGFAMA
MUFUMUFCMU
FAMCFAMGFU
FCMAFUMCESA
MGFGMUFGMG
M
SUF-OH-3′
FAMSCMSAM-
FAMAFUMGFG
MCFUMGFCMA
MCFUMGFCMA
FGMUFCMUFSG
MUFCMGFUMU
M
SUF-OH-3′
FCMCFAMUFG
MCFAMGFGMA
MAFUMCFCMU
FUMCFAMUFG
FGMUFCMGFSA
MGFUMGFGMC
M
SUF-OH-3′
FCMSUMSGM-
FCMGFAMUFC
MGFAMAFGMA
MAFUMCFUMU
FUMGFAMUFC
FCMAFGMCFSA
MGFAMCFAMG
M
SUF-OH-3′
FGMSAMSUM-
FUMGFUMUFC
MGFCMAFGMA
MUFUMCFUMG
FAMGFAMAFC
FCMCFAMAFSC
MAFGMGFAMA
MUF-OH-3′
FCMSAMSGM-
FUMCFUMUFC
MUFUMGFGMC
MUFGMCFCMA
FAMGFAMAFG
FAMCFUMCFSU
MAFAMCFAMG
M
SUF-OH-3′
FGMSAMSAM-
FUMCFUMUFC
MGFAMGFGMG
MAFCMCFCMUF
FUMGFAMAFG
MAFGMUFUMG
SAF-OH-3′
FGMSCMSAM-
FCMAFGMGFU
MUFGMUFAMA
MUFUMUFAMC
FAMAFCMCFU
FAMUFCMAFSC
MGFCMCFCMCF
M
SUF-OH-3′
FUMUFUMAFC
MAFGMUFGMA
MAFUMCFAMC
FUMGFUMAFA
FUMGFGMAFSA
MAFAMCFCMU
M
SUF-OH-3′
FAMCFAMUFC
MUFCMCFAMG
MAFCMUFGMG
FUMGFAMUFG
FAMAFUMCFSU
MUFAMAFAMA
M
SUF-OH-3′
FCMSCMSUM-
FUMGFGMAFA
MGFGMAFAMG
MUFCMUFUMC
FAMUFUMCFC
FCMAFAMAFSU
MAFGMUFGMA
M
SUF-OH-3′
FCMUFUMCFCM
MGFAMAFUMU
MGFAMUFUMC
M
SUF-OH-3′
FUMGFAMGFU
MCFGMCFAMG
MGFCMUFGMC
FCMAFCMUFCM
FGMGFCMCFSA
M
SUF-OH-3′
FCMGFGMAFG
MAFUMGFCMC
MUFGMGFCMA
FAMCFUMCFCM
FUMCFUMCESA
M
SAF-OH-3′
FAMUFUMAFC
MGFUMAFGMG
MUFCMCFUMA
FAMGFUMAFA
FCMGFGMUFSU
MUFCMCFGMA
M
SUF-OH-3′
FGMSUMSUM-
FAMCFGMGFU
MGFGMCFGMA
MUFUMCFGMC
FAMAFCMCFG
FCMUFAMCFSA
MUFAMGFGMA
M
SUF-OH-3′
FGMSUMSAM-
FGMUFUMUFC
MGFUMAFGMG
MGFCMCFUMA
FCMGFAMAFA
FCMAFUMCFSC
MCFCMGFUMA
M
SUF-OH-3′
FGMUFGMUFC
MAFUMAFGMA
MAFUMCFUMA
FUMGFAMCFA
FUMGFUMGESA
MCFCMGFCMUF
M
SUF-OH-3′
FUMCFAMUFC
MCFAMCFAMU
MUFAMUFGMU
FAMGFAMUFG
FGMAFUMCFSU
MAFCMAFCMCF
M
SUF-OH-3′
FUMGFUMGFA
MGFCMAFAMG
MUFCMUFUMG
FAMUFCMAFC
FCMGFGMAFSA
MAFUMAFGMA
M
SAF-OH-3′
FCMCFAMAFA
MUFUMUFGMA
MCFUMCFAMA
FGMUFUMUFG
FAMCFCMAFSA
MGFGMAFUMU
M
SAF-OH-3′
FUMSUMSGM-
FUMUFGMAFU
MUFCMUFUMC
MUFGMAFAMG
FAMAFUMCFA
FAMUFGMUFSA
MAFCMAFGMC
M
SUF-OH-3′
FAMSAMSCM-
FUMAFUMAFA
MUFAMGFGMU
MAFAMCFCMU
FAMUFUMUFSA
MAFAMAFCMC
M
SUF-OH-3′
FGMSGMSAM-
FUMAFGMUFA
MAFAMAFCMA
MUFUMGFUMU
FAMUFAMCFU
FUMGFCMUFSU
MAFUMGFUMA
SUF-OH-3′
FCMCFAMUFCM
MGFAMGFGMC
FUMGFAMUFG
MGFUMCFAMA
M
SUF-OH-3′
FGMUFAMGFC
MUFCMCFUMU
MUFAMAFGMG
FAMGFCMUFA
FAMAFCMUFSU
MCFUMUFCMU
M
SUF-OH-3′
FUMSUMSAM-
FGMAFAMCFU
MAFUMGFUMA
MUFUMAFCMA
FAMAFGMUFU
FUMCFCMUFSA
MCFCMUFUMA
M
SAF-OH-3′
FGMSCMSUM-
FGMUFGMGFA
MUFCMUFUMA
MCFUMAFAMG
FGMUFCMCFA
FAMUFGMCFSA
MCFAMCFAMG
M
SUF-OH-3′
FUMSUMSGM-
FUMUFUMGFG
MGFCMCFCMGF
MCFCMGFGMG
FCMAFGMAFSA
M
SAF-OH-3′
FAMCFUMCFCM
MCFUMCFAMG
FCMGFGMAFG
MUFUMUFCMU
M
SAF-OH-3′
MCFUMGFCMU
FCMAFGMCFG
MGFAMGFUMU
M
SUF-OH-3′
FUMSCMSUM-
The 3′ terminus of the sense strand of certain compounds was conjugated to a long chain fatty acid (LCFA) domain or “uptake motif” which improves the uptake of nucleic acid compounds into cells both in vitro and in vivo (International Patent Application Publication No. WO 2019/232255). The conjugated compounds are shown in Table 4. “Start” and “End” correspond to the 5′ and 3′ nucleotide positions of the nucleotide sequence of the human PMP22 mRNA (NCBI Reference Sequence NM_000304.4, deposited with GenBank on Nov. 22, 2018; SEQ ID NO: 1170) to which the nucleotides of the antisense strand are complementary. Each row represents a sense and antisense strand pair of an siRNA. The nucleotide sequences for both the modified and unmodified sense and antisense strands are included.
Conjugated compounds were formed as in the structures below, where the nucleotide shown is the 3′ terminal nucleotide, “B” is nucleobase and “R” is the substituent at the 2′ carbon of the nucleoside sugar.
The uptake motif DTx-01-08 was conjugated to the sense strand, using the “C7OH” linker
attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand via the phosphate group to form the conjugate group named “C7OH-[DTx-01-08] in Table 4.
The uptake motif DTx-01-32 was conjugated to the sense strand, using the “C7OH” linker
attached to the 3′ carbon of the 3′ terminal nucleotide of the sense strand via the phosphate group to form the conjugate group named “C7OH-[DTx-01-32] in Table 4.
In Table 4 and elsewhere herein, modified sugar moieties are indicated by a subscript notation following the nucleotide, and modified internucleotide linkages are indicated by a superscript notation. 5′ and 3′ terminal groups are also indicated. A nucleotide followed by the subscript “F” is a 2′-fluoro nucleotide; a nucleotide followed by the subscript “M” is a 2′-O-methyl nucleotide; a nucleotide followed by the subscript “E” is a 2′-O-methoxyethyl nucleotide; and a nucleotide followed by the subscript “D” is a beta-D-deoxyribonucleotide. The nucleobase of each “CE” nucleotide is a 5-methylcytosine; each other “C” is a non-methylated cytosine; the nucleobase of each “UE” nucleotide is a 5-methyluracil; each other “U” is a non-methylated uridine. A superscript “S” is a phosphorothioate internucleotide linkage; all other internucleotide linkages are phosphodiester internucleotide linkages. For example, “UFSCM” is a 2′-flourouridine linked to a 2′-O-methylcytidine by a phosphorothioate internucleotide linkage. “GMUF” is a 2-O-methylguanosine linked to a 2′-fluorouridine by a phosphodiester internucleotide linkage. A hydroxyl group is at the 5′ carbon of the 5′ terminal nucleotide is indicated by “5′-OH”; a phosphate group at the 5′ carbon of the 5′ terminal nucleotide is indicated by “5′-PO4”; a 5′-VP modification at the 5′ terminal nucleotide of an antisense strand is indicated by “5′-VP”; and a hydroxyl group at the 3′ carbon of the 3′ terminal nucleotide is indicated by “OH-3′.”
MUFGMGFCMAFG
MAFAMCFUMGFSU
MCFAMGFAMGF
M
SAF-C7OH-
MUFCMCFUMGFU
MUFCMUFUMCFSU
MGFAMAFCMAF
M
SGF-C7OH-
MGFUMCFUMGFU
MGFCMGFUMGFSA
MAFCMCFAMGF
M
SUF-C7OH-
MUFCMCFUMGFU
MUFCMUFUMCFSU
MGFAMAFCMAF
M
SGF-C7OH-
MCFAMUFCMUFU
MCFAMGFCMAFSU
MUFGMAFUMCF
M
SUF-C7OH-
MGFGMAFAMUFC
MUFUMCFCMAFSA
MCFCMAFGMUF
M
SAF-C7OH-
MUFGMCFUMGFA
MGFUMAFUMCFSA
MCFAMAFCMAF
M
SUF-C7OH-
MUFCMAFCMCFAM
MGFAMUFGMAF
SUF-C7OH-
MUFUMCFUMGFU
MCFUMCFUMGFSU
MAFAMUFGMCF
M
SUF-C7OH-
MUFUMCFCMUFG
MUFUMCFUMUFSC
MAFAMCFAMGF
M
SUF-C7OH-
MGFAMAFUMCFU
MUFCMCFAMAFSA
MUFCMCFAMGF
M
SUF-C7OH-
MCFGMGFUMGFU
MCFAMUFCMUFSA
MCFGMCFUMGF
M
SUF-C7OH-
MUFGMCFUMGFA
MGFUMAFUMCFSA
MCFAMAFCMAF
M
SUF-C7OH-
MGFUMUFGMCFU
MGFAMGFUMAFU
MCFAMAFCMAF
MCFSAMSUF-
MGFUMUFGMCFU
MGFAMGFUMAFU
MCFAMAFCMAF
MCFSAMSUF-
MCFCMUFGMUFU
MGFCMUFGMAFSG
MGFGMAFGMGF
M
SUF-C7OH-
MUFGMUFUMGFC
MUFGMAFGMUFSA
MCFAMGFGMAF
M
SUF-C7OH-
MGFUMUFGMCFU
MGFAMGFUMAFSU
MAFCMAFGMGF
M
SCF-C7OH-
MGFCMUFGMAFG
MUFAMUFCMAFSU
MGFCMAFAMCF
M
SCF-C7OH-
MAFGMUFAMUFC
MAFUMCFGMUFSC
MCFUMCFAMGF
M
SCF-C7OH-
MAFCMAFCMGFCM
MGFUMCFCMAF
SUF-C7OH-
MUFGMAFUMCFC
MUFGMUFCMGFSA
MCFAMUFGMGF
M
SUF-C7OH-
MUFCMAFUMCFU
MUFCMAFGMCFSA
MGFAMUFCMGF
M
SUF-C7OH-
MUFCMUFUMCFU
MGFCMCFAMAFSC
MGFAMAFCMAF
M
SUF-C7OH-
MGFUMUFGMCFU
MGFAMGFUMAFU
MCFAMAFCMAF
MCFSAMSUF-
MUFCMAFCMUFG
MGFAMAFUMCFSU
MGFAMUFGMUF
MUFCMAFCMUFG
MGFAMAFUMCFSU
MGFAMUFGMUF
M
SUF-C7OH-
MUFCMAFCMCFCM
MGFAMAFGMAF
SAF-C7OH-
MGFUMUFUMUFA
MCFAMUFCMAFSC
MAFCMCFUMGF
M
SUF-C7OH-
MAFCMAFUMCFA
MCFUMGFGMAFSA
MGFUMAFAMAF
M
SUF-C7OH-
MAFAMUFCMUFU
MCFCMAFAMAFSU
MUFUMCFCMAF
M
SUF-C7OH-
MCFCMAFAMAFU
MUFCMUFUMGFSC
MGFGMAFAMGF
M
SUF-C7OH-
MCFUMUFGMCFU
MGFGMUFCMUFSG
MAFGMAFAMUF
M
SUF-C7OH-
MCFUMGFUMGFC
MGFUMGFAMUFSG
MAFGMAFCMCF
M
SAF-C7OH-
MGFUMGFCMUFG
MCFGMGFCMCFSA
MAFCMUFCMAF
M
SUF-C7OH-
MCFCMCFGMGFAM
MGFGMUFGMCF
SUF-C7OH-
MAFGMUFGMGFC
MAFUMCFUMCESA
MCFUMCFCMGF
M
SAF-C7OH-
MCFGMGFAMUFU
MAFCMUFCMCFSU
MCFGMAFGMUF
M
SAF-C7OH-
MAFCMUFCMCFUM
MGFUMAFAMUF
SUF-C7OH-
MGFUMUFUMCFG
MCFCMUFAMCESA
MAFCMCFGMUF
M
SUF-C7OH-
MUFCMGFCMCFUM
MGFAMAFAMCF
SUF-C7OH-
MUFCMAFGMCFG
MGFUMGFUMCFSA
MGFAMGFAMAF
M
SUF-C7OH-
MUFCMAFUMCFU
MAFUMGFUMGFSA
MGFAMCFAMCF
M
SUF-C7OH-
MUFCMUFAMUFG
MUFGMAFUMCFSU
MGFAMUFGMAF
M
SUF-C7OH-
MGFAMUFCMUFU
MGFCMGFGMAFSA
MUFCMAFCMAF
M
SAF-C7OH-
MGFGMGFAMAFA
MAFCMAFGMAFSA
MCFCMUFUMCFC
M
SAF-C7OH-
MUFCMSTDSTD-
MAFUMCFCMCFAM
MAFUMUFUMUF
SAF-C7OH-
MAFAMCFUMCFA
MAFAMCFCMAFSA
MUFUMUFGMGF
M
SAF-C7OH-
MGFAMUFUMGFA
MAFGMAFUMGFSU
MUFCMAFAMCF
M
SAF-C7OH-
MAFUMUFGMAFA
MGFAMUFGMUFSA
MAFUMCFAMAF
M
SUF-C7OH-
MAFAMAFAMCFC
MUFAMUFUMUFSA
MUFUMAFUMAF
M
SUF-C7OH-
MUFAMUFUMGFU
MUFUMGFCMUFSU
MUFAMCFUMAF
M
SUF-C7OH-
MUFCMAFGMCFCM
MGFAMUFGMGF
SUF-C7OH-
MAFAMGFAMAFG
MUFAMGFCMUFSA
MUFUMUFAMAF
M
SAF-C7OH-
MGFCMUFAMAFG
MGFAMAFCMUFSU
MGFCMUFAMCF
M
SUF-C7OH-
MCFUMUFUMAFC
MAFUMCFCMUFSA
MAFGMUFUMCF
M
SAF-C7OH-
MGFAMCFUMAFA
MGFAMUFGMCFSA
MUFCMCFAMCF
M
SUF-C7OH-
MGFGMGFAMAFA
MAFCMAFGMAFSA
MCFCMUFUMCFC
M
SAF-C7OH-
MUFCMSTDSTD-
MAFUMCFCMCFAM
MAFUMUFUMUF
SAF-C7OH-
MAFAMCFUMCFA
MAFAMCFCMAFSA
MUFUMUFGMGF
M
SAF-C7OH-
MGFAMUFUMGFA
MAFGMAFUMGFSU
MUFCMAFAMCE
M
SAF-C7OH-
MAFUMUFGMAFA
MGFAMUFGMUFSA
MAFUMCFAMAF
M
SUF-C7OH-
MAFAMAFAMCFC
MUFAMUFUMUFSA
MUFUMAFUMAF
M
SUF-C7OH-
MUFAMUFUMGFU
MUFUMGFCMUFSU
MUFAMCFUMAF
M
SUF-C7OH-
MUFCMAFGMCFCM
MGFAMUFGMGF
SUF-C7OH-
MAFAMGFAMAFG
MUFAMGFCMUFSA
MUFUMUFAMAF
M
SAF-C7OH-
MGFCMUFAMAFG
MGFAMAFCMUFSU
MGFCMUFAMCF
M
SUF-C7OH-
MCFUMUFUMAFC
MAFUMCFCMUFSA
MAFGMUFUMCE
M
SAF-C7OH-
MGFAMCFUMAFA
MGFAMUFGMCESA
MUFCMCFAMCF
M
SUF-C7OH-
MGFGMGFAMAFA
MAFCMAFGMAFSA
MCFCMUFUMCFC
M
SAF-C7OH-
MUFCMSTDSTD-
MAFUMCFCMCFAM
MAFUMUFUMUF
SAF-C7OH-
MGFAMUFUMGFA
MAFGMAFUMGFSU
MUFCMAFAMCF
M
SAF-C7OH-
MAFAMGFAMAFG
MUFAMGFCMUFSA
MUFUMUFAMAF
M
SAF-C7OH-
MAFAMCFUMCFA
MAFAMCFCMAFSA
MUFUMUFGMGF
MAFUMUFGMAFA
MGFAMUFGMUFSA
MAFUMCFAMAF
M
SUF-C7OH-
MAFAMAFAMCFC
MUFAMUFUMUFSA
MUFUMAFUMAF
M
SUF-C7OH-
MUFAMUFUMGFU
MUFUMGFCMUFSU
MUFAMCFUMAF
M
SUF-C7OH-
MUFCMAFGMCFCM
MGFAMUFGMGF
SUF-C7OH-
MGFCMUFAMAFG
MGFAMAFCMUFSU
MGFCMUFAMCF
M
SUF-C7OH-
MCFUMUFUMAFC
MAFUMCFCMUFSA
MAFGMUFUMCF
M
SAF-C7OH-
MGFAMCFUMAFA
MGFAMUFGMCFSA
MUFCMCFAMCF
M
SUF-C7OH-
MGFUMUFCMCFU
MGFUMUFCMUFSU
MAFCMAFGMAF
M
SCF-C7OH-
MUFUMAFUMAFA
MCFAMCFUMUFSU
MAFAMAFUMAF
M
SUF-C7OH-
MAFAMUFAMAFA
MUFCMUFCMAFSA
MUFUMUFAMUF
M
SAF-C7OH-
MGFUMUFGMAFA
MUFCMUFUMAFSA
MAFCMAFCMGF
M
SAF-C7OH-
MAFCMUFGMUFA
MGFAMUFGMUFSA
MGFUMUFGMGF
M
SUF-C7OH-
MAFGMGFCMCFA
MCFCMAFUMGFSA
MCFUMGFGMAF
M
SUF-C7OH-
MAFCMUFGMUFG
MUFGMGFAMCFSU
MGFUMUFGMGF
M
SAF-C7OH-
MAFUMUFUMAFU
MAFAMCFAMCFSU
MAFUMAFGMGF
M
SUF-C7OH-
MUFUMGFCMUFG
MAFGMUFAMUFSC
MAFAMCFAMGF
M
SAF-C7OH-
MCFUMGFAMGFU
MAFUMCFAMUFSC
MAFGMCFAMAF
M
SGF-C7OH-
MGFAMGFUMAFU
MCFAMUFCMGFSU
MUFCMAFGMCF
M
SCF-C7OH-
MUFCMGFUMCFCM
MGFAMUFGMAF
SUF-C7OH-
MUFGMCFUMGFG
MUFGMCFUMGFSC
MCFAMCFCMGFC
M
SUF-C7OH-
MGFAMSTDSTD-
MCFUMGFCMUFG
MUFUMCFGMUFSC
MAFGMCFAMCF
M
SUF-C7OH-
MGFCMAFAMCFU
MGFAMUFCMUFSC
MGFCMGFUMGF
M
SUF-C7OH-
MUFCMCFAMCFCM
MGFAMCFAMUF
SUF-C7OH-
MUFGMUFUMUFC
MUFCMAFUMCFSA
M
SUF-C7OH-
MGFGMCFUMGFC
MAFGMUFCMUFSG
MCFCMAFUMUF
M
SUF-C7OH-
MUFCMUFGMCFCM
MGFAMAFGMAF
SUF-C7OH-
MGFGMCFCMGFG
MGFCMAFGMAFSA
MCFCMAFAMAF
M
SAF-C7OH-
MCFCMGFCMUFGM
MGFGMAFGMUF
SAF-C7OH-
MCFUMGFAMGFC
MAFGMAFAMCFSU
MAFGMCFGMGF
M
SUF-C7OH-
MCFUMGFAMGFU
MAFUMCFAMUFSC
MAFGMCFAMAF
M
SGF-C7OH-
MGFAMGFUMAFU
MCFAMUFCMGFSU
MUFCMAFGMCF
M
SCF-C7OH-
MUFCMGFUMCFCM
MGFAMUFGMAF
SUF-C7OH-
MUFGMCFUMGFG
MUFGMCFUMGFSC
MCFAMCFCMGFC
M
SUF-C7OH-
MGFAMSTDSTD-
MCFUMGFCMUFG
MUFUMCFGMUFSC
MAFGMCFAMCF
M
SUF-C7OH-
MUFCMAFCMCFCM
MGFAMAFGMAF
SAF-C7OH-
MGFUMUFUMUFA
MCFAMUFCMAFSC
MAFCMCFUMGF
M
SUF-C7OH-
MAFCMAFUMCFA
MCFUMGFGMAFSA
MGFUMAFAMAF
M
SUF-C7OH-
MUFCMAFCMUFG
MGFAMAFUMCFSU
MGFAMUFGMUF
M
SUF-C7OH-
MAFAMUFCMUFU
MCFCMAFAMAFSU
MUFUMCFCMAF
M
SUF-C7OH-
MCFCMAFAMAFU
MUFCMUFUMGFSC
MGFGMAFAMGF
M
SUF-C7OH-
MGFUMGFCMUFG
MCFGMGFCMCFSA
MAFCMUFCMAF
M
SUF-C7OH-
MAFGMUFGMGFC
MAFUMCFUMCFSA
MCFUMCFCMGF
M
SAF-C7OH-
MAFCMUFCMCFUM
MGFUMAFAMUF
SUF-C7OH-
MGFUMUFUMCFG
MCFCMUFAMCESA
MAFCMCFGMUF
M
SUF-C7OH-
MUFCMGFCMCFUM
MGFAMAFAMCF
SUF-C7OH-
MUFCMAFUMCFU
MAFUMGFUMGFSA
MGFAMCFAMCF
M
SUF-C7OH-
MUFCMUFAMUFG
MUFGMAFUMCFSU
MGFAMUFGMAF
M
SUF-C7OH-
MGFAMUFCMUFU
MGFCMGFGMAFSA
MUFCMAFCMAF
M
SAF-C7OH-
MAFAMGFGMGFA
MAFAMAFCMAFG
MCFCMUFUMCFC
MAFSAMSAF-
MUFCMCFCMSUM
SUM-OH-3′
MAFAMAFUMCFC
MCFAMAFAMCFU
MAFUMUFUMUF
MCFAMAF-C7OH-
MUFUMGFAMUFU
MGFAMAFGMAFU
MUFCMAFAMCF
MGFSUMSAF-
MUFAMAFAMGFA
MAFGMUFAMGFC
MUFUMUFAMAF
MUFSAMSAF-
MAFAMGFGMGFA
MAFAMAFCMAFG
MCFCMUFUMCFC
MAFSAMSAF-
MUFCMCFCMSUM
SUM-OH-3′
MAFAMAFUMCFC
MCFAMAFAMCFU
MAFUMUFUMUF
MCFAMAF-C7OH-
MUFUMGFAMUFU
MGFAMAFGMAFU
MUFCMAFAMCF
MGFSUMSAF-
MUFAMAFAMGFA
MAFGMUFAMGFC
MUFUMUFAMAF
MUFSAMSAF-
MCFCMUFAMAFCM
MGFGMAFUMGF
SAF-C7OH-
MAFGMAFAMAFU
MAFAMGFAMUFSA
MCFUMGFGMGF
M
SAF-C7OH-
MGFCMAFUMUFU
MUFCMUFGMAFSU
MGFCMAFAMAF
M
SUF-C7OH-
MUFGMUFGMGFA
MCFUMAFAMGESA
MCFAMCFAMGF
M
SUF-C7OH-
MAFUMCFCMUFA
MAFCMAFGMUFA
MGFGMAFUMGF
MUFSAMSAF-
MCFCMAFGMAFA
MAFUMAFAMGFA
MCFUMGFGMGF
MUFSAMSAF-
MCFAMAFCMUFG
MUFAMGFAMUFG
MGFUMUFGMGF
MUFSAMSUF-
MUFUMGFCMAFU
MUFUMUFCMUFG
MGFCMAFAMAF
MAFSUMSUF-
MUFGMUFGMUFG
MGFAMCFUMAFA
MCFAMCFAMGF
MAFUMCFCMUFA
MAFCMAFGMUFA
MGFGMAFUMGF
MUFSAMSAF-
MCFCMAFGMAFA
MAFUMAFAMGFA
MCFUMGFGMGF
MUFSAMSAF-
MCFAMAFCMUFG
MUFAMGFAMUFG
MGFUMUFGMGF
MUFSAMSUF-
MUFUMGFCMAFU
MUFUMUFCMUFG
MGFCMAFAMAF
MAFSUMSUF-
MUFGMUFGMUFG
MGFAMCFUMAFA
MCFAMCFAMGF
MGFSAMSUF-
MGFUMUFGMCFUF
MCFAMAFCMAF
MGFUMUFGFCFUM
MCMAMAFCMAF
MGFUMUFGMCFU
MGFAMGFUMAFU
MCFAMAFCMAF
MCMSAMSUM-
MCMAMAFCMAF
MAFCMUMCMAM
MUMCMSAMSUM-
FGMGMAMGMGM
SAMSGM-OH-3′
MAFCMUMCMAM
MUMCMAMUM-
SAMSGM-OH-3′
MGFUMUFGMCFU
MGFAMGFUMAFU
MCFAMAFCMAF
MCMSAMSUM-
MGFUMUFGMCFU
MGFAMGFUMAFU
MCFAMAFCMAF
MCMSAMSUM-
MGFUMUFGMCFU
MGFAMGFUMAFU
MCFAMAFCMAF
MCESAESUM-
MCFAMAFCMAF
MUFGMCFUMGFA
MGFUMAFUMCFA
MAFGMCFAMAF
MUFSCMSGF-
MCFUMGFAMGFU
MAFUMCFAMUFC
MUFCMAFGMCF
MGFSUMSCF-
MCFUMCFAMGF
MGFGMAFCMAFC
MGFCMAFAMCFU
MGFUMCFCMAF
MGFSAMSUF-
MCFAMUFGMAFU
MCFCMUFGMUFCM
MCFAMUFGMGF
MGFAMUFCMAFU
MCFUMUFCMAFG
MGFAMUFCMGF
MCFSAMSUF-
MGFUMUFGMCFU
MGFGMGFUMAFU
MCFAMAFCMAF
MCFSAMSUF-
MCFUMCFCMUFGM
MGFGMAFGMGF
MCFCMUFGMUFU
MGFCMUFGMAFG
MCFAMGFGMAF
MUFSAMSUF-
MCFUMGFUMUFG
MCFUMGFAMGFU
MAFCMAFGMGF
MAFSUMSCF-
MUFGMUFUMGFC
MUFGMAFGMUFA
MAFAMCFAMGF
MUFSCMSAF-
MUFUMGFCMUFG
MAFGMUFAMUFC
MGFCMAFAMCF
MAFSUMSCF-
MUFGMCFUMGFA
MGFUMAFUMCFA
MAFGMCFAMAF
MUFSCMSGF-
MCFUMGFAMGFU
MAFUMCFAMUFC
MUFCMAFGMCF
MGFSUMSCF-
MCFUMCFAMGF
MCFAMUFCMGFU
MCFCMUFCMCFAM
MGFAMUFGMAF
MGFGMUFGMCFU
MGFGMUFGMCFU
MCFAMCFCMGFC
MGFSCMSUF-
MGFAMCFGMSU
M
SGM-OH-3′
MUFGMCFUMGFC
MUFGMUFUMCFG
MAFGMCFAMCF
MUFSCMSUF-
MGFGMAFCMAFC
MGFCMAFAMCFU
MGFUMCFCMAF
MGFSAMSUF-
MAFCMGFCMAFA
MCFUMGFAMUFC
MGFCMGFUMGF
MUFSCMSUF-
MUFGMUFCMCFA
MCFCMAFCMUFGM
MGFAMCFAMUF
MAFCMUFGMUFU
MUFCMUFCMAFU
MCFAMGFUMGF
MCFSAMSUF-
MAFUMGFGMCFU
MGFCMAFGMUFC
MCFCMAFUMUF
MUFSGMSUF-
MCFAMUFGMAFU
MCFCMUFGMUFCM
MCFAMUFGMGF
MGFAMUFCMAFU
MCFUMUFCMAFG
MGFAMUFCMGF
MCFSAMSUF-
MGFUMUFCMUFU
MCFUMGFCMCFAM
MGFAMAFCMAF
MCFUMUFCMUFG
MCFCMAFAMCFUM
MGFAMAFGMAF
MCFUMUFCMAFCM
MGFAMAFGMAF
MAFGMGFUMUFU
MUFAMCFAMUFC
MAFCMCFUMGF
MAFSCMSUF-
M
SUM-OH-3′
MUFUMAFCMAFU
MCFAMCFUMGFG
MGFUMAFAMAF
MAFSAMSUF-
MCFAMUFCMAFCM
MGFAMUFGMUF
MGFGMAFAMUFC
MUFUMCFCMAFA
MUFUMCFCMAF
MAFSUMSUF-
MUFUMCFCMAFA
MAFUMUFCMUFU
MGFGMAFAMGF
MGFSCMSUF-
MGFAMGFUMGFC
MUFGMCFGMGFC
MAFCMUFCMAF
MCFSAMSUF-
MGFGMAFGMUFG
MGFCMAFUMCFU
MCFUMCFCMGF
MCFSAMSAF-
MUFUMAFCMUFC
MCFUMAFCMGFG
MGFUMAFAMUF
MUFSUMSUF-
MCFGMGFUMUFU
MCFGMCFCMUFAM
MAFCMCFGMUF
MUFUMUFCMGFC
MCFUMAFCMAFU
MGFAMAFAMCF
MCFSCMSUF-
MUFGMUFCMAFU
MCFUMAFUMGFU
MGFAMCFAMCF
MGFSAMSUF-
MCFAMUFCMUFA
MUFGMUFGMAFU
MGFAMUFGMAF
MCFSUMSUF-
M
SUM-OH-3′
MGFUMGFAMUFC
MUFUMGFCMGFG
MUFCMAFCMAF
MAFSAMSAF-
MCFAMAFAMCFU
MCFAMAFAMCFCM
MUFUMUFGMGF
MUFGMAFUMUFG
MAFAMGFAMUFG
MAFUMCFAMAF
MUFSAMSUF-
MAFUMAFAMAFA
MCFCMUFAMUFU
MUFUMAFUMAF
MUFSAMSUF-
MAFGMUFAMUFU
MGFUMUFUMGFC
MUFAMCFUMAF
MUFSUMSUF-
MCFAMUFCMAFG
MCFCMUFCMGFUM
MGFAMUFGMGF
MUFAMGFCMUFA
MAFGMGFAMAFC
MGFCMUFAMCF
MUFSUMSUF-
MAFAMCFUMUFU
MAFCMAFUMCFCM
MAFGMUFUMCF
MUFGMGFAMCFU
MAFAMGFAMUFG
MUFCMCFAMCE
MCFSAMSUF-
MUFUMGFGMCFC
MGFGMGFCMAFG
MCFCMAFAMAF
MAFSAMSAF-
MCFUMCFCMGFCM
MGFGMAFGMUF
MCFGMCFUMGFA
MGFCMAFGMAFA
MAFGMCFGMGF
MCFSUMSUF-
MGFGMUFGMCFU
MGFGMUFGMCFU
MCFAMCFCMGFC
MGFSCMSUF-
MGFAMCFGMSU
M
SGM-OH-3′
MUFGMCFUMGFC
MUFGMUFUMCFG
MAFGMCFAMCF
MUFSCMSUF-
MUFGMUFCMCFA
MCFCMAFCMUFGM
MGFAMCFAMUF
MAFCMUFGMUFU
MUFCMUFCMAFU
MCFAMGFUMGF
MCFSAMSUF-
MAFUMGFGMCFU
MGFCMAFGMUFC
MCFCMAFUMUF
MUFSGMSUF-
MCFUMUFCMUFG
MCFCMAFAMCFUM
MGFAMAFGMAF
MCFUMUFCMAFCM
MGFAMAFGMAF
MAFGMGFUMUFU
MUFAMCFAMUFC
MAFCMCFUMGF
MAFSCMSUF-
M
SUM-OH-3′
MUFUMAFCMAFU
MCFAMCFUMGFG
MGFUMAFAMAF
MAFSAMSUF-
MCFAMUFCMAFCM
MGFAMUFGMUF
MGFAMGFUMGFC
MUFGMCFGMGFC
MAFCMUFCMAF
MCFSAMSUF-
MUFUMAFCMUFC
MCFUMAFCMGFG
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Unconjugated compounds were tested for their ability to inhibit the expression of PMP22 in human Schwann cells that express endogenous PMP22 and HEK cells engineered to express human PMP22 (HEK-PMP22 cells). Transfection experiments and PMP22 quantitation were performed according to the methods described herein.
Schwann cells and HEK-PMP22 cells were transfected with siRNAs at doses of 0.3 nM, 3 nM, and 30 nM. RNA was isolated 48 hours later, reverse transcribed to cDNA and PMP22 expression was quantified by qPCR. The average PMP22 expression for each of four replicates was calculated and shown in Tables 5 through 10. Several of the siRNAs inhibited PMP22 expression in a dose-dependent manner.
Schwann cells and HEK-PMP22 cells were transfected with siRNAs at doses of 3 nM and 30 nM. RNA was isolated 48 hours later, reverse transcribed to cDNA and PMP22 expression was quantified by PCR. The average PMP22 expression for each of four replicates was calculated and shown in Tables 11 and 12. Several of the siRNAs inhibited PMP22 expression in a dose-dependent manner.
Compounds DT-000904 through DT-000928 target the 3′-UTR of human PMP22. As HEK-PMP22 cells do not express the 3′-UTR of PMP22, these compounds were tested in Schwann cells only.
Compounds DT-001010 through DT-001034 target the 5′-UTR of human PMP22. As HEK-PMP22 cells do not express the 5′-UTR of PMP22, these compounds were tested in Schwann cells only.
Certain compounds were selected for additional testing in a dose-response experiment. Schwann cells and HEK-PMP22 cells were transfected with siRNAs at doses of 0.3 nM, 1 nM, 3 nM, 10 nM and 30 nM. RNA was isolated 48 hours later, reverse transcribed to cDNA and PMP22 expression was quantified by qPCR. The average PMP22 expression for each of four replicates was calculated and shown in Tables 15 through 18. Several of the siRNAs inhibited PMP22 expression in a dose-dependent manner.
Based on transfection data, certain compounds were identified as “hits” and selected for conjugation. Table 19 illustrates the parent unconjugated siRNAs identified as “hits” and the one or more conjugated siRNAs derived therefrom. Also shown are the lengths of the sense strand, the uptake motif attached to the sense strand, and the 5′ terminal moiety of the antisense strand.
Conjugated compounds were tested for their ability to inhibit the expression of PMP22 in HER cells engineered to express human PMP22 (HEK-PMP22 cells). These studies were performed under free uptake conditions as described herein. The “parent” unconjugated compound ID is indicated next to each conjugated compound ID.
Schwann cells and HEK-PMP22 cells were treated with siRNAs as indicated in the Tables below. RNA was isolated 48 hours later, reverse transcribed to cDNA and PMP22 expression was quantified by qPCR. The average PMP22 expression for each of four
Conjugated PMP22 siRNAs were tested in wild-type C57BL/6J mice. In this experiment, control siRNAs were DT-000155 and DT-000337, both DTx-01-08-conjugated siRNAs targeting PTEN, each having a unique nucleotide sequence. Also tested was DT-000428, a fully phosphorothioated LNA gapmer antisense oligonucleotide (ASO) targeting PMP22, where a 10-nucleotide DNA gap is flanked by 3-nucleotide LNA wings (5′-ALTLCLTDTDCDADADTDCDADADCDALGLCL-3′; subscript L is an LNA nucleotide and subscript D is a beta-D-deoxyribonucleotide; nucleotides four to 19 of SEQ ID NO: 591). Groups of five mice each were treated with PBS or compound at a dose of 30 mg/kg according to the dosing schedule indicated in Table 35. On Day 12, mice were sacrificed, and RNA was collected from tissue for RNA extraction and quantitation of mouse PMP22 mRNA levels by quantitative RT-PCR. The average percent expression in the central sciatic nerve was calculated for each treatment and is shown in Table 35.
C3-PMP22 mice express three to four copies of a wild-type human PMP22 gene and are used as an experimental model of CMT1A. Conjugated siRNAs targeted to human PMP22 were selected for their ability to reduce human PMP22 in C3-PMP22 mice. Experiments were performed as described herein.
In this experiment, the control siRNA was DT-000337, a DTx-01-08-conjugated siRNA targeting PTEN. Also tested was DT-000428, a fully phosphorothioated LNA gapmer antisense oligonucleotide targeting PMP22, where a 10-nucleotide DNA gap is flanked by 3-nucleotide LNA wings (5′-ALTLCLTDTDCDADADTDCDADADCDALGLCL-3′; nucleotides 4 to 19 of SEQ ID NO: 438; subscript L is an LNA nucleotide and subscript D is a beta-D-deoxyribonucleotide). Groups of six mice each were treated with PBS, siRNA compound at a dose of 50 mg/kg, or DT-000428 at a dose of 100 mg/kg on Days 1, 7, and 14. On Day 21, mice were sacrificed, and RNA was collected from tissue for RNA extraction and quantitation of human PMP22 mRNA levels by quantitative RT-PCR. The average percent expression in the sciatic nerve and tibial nerve was calculated for each treatment and is shown in Table 36.
The most active compound from the above study, DT-000623, was further tested. Groups of six C3-PMP22 mice each were treated with PBS or DT-000623 siRNA compound for a total of 1 dose, 2 doses, or 3 doses, at the dosing schedule indicated in Table 37. For comparison, wild-type mice were treated with PBS on the same dosing schedule. After 21 days, mice were sacrificed, and RNA was collected from tissue for RNA extraction and quantitation of human PMP22 mRNA levels by quantitative RT-PCR. mRNA levels for the mouse sciatic nerve markers MPZ, Pou3F1, Sc5d, and Id2 were also calculated. The average percent expression for each mRNA in the sciatic nerve and tibial nerve was calculated for each treatment and is shown in Table 37. In each table, wild-type PBS indicates data collected from wild-type mice treated with PBS. All other data were obtained in C3-PMP22 mice.
DT-000623 and variants, DT-000811 and DT-000812, were tested in C3-PMP22 mice. Groups of five C3-PMP22 mice each were treated with PBS or a single dose of 10 mg/kg, 30 mg/kg, or 100 mg/kg of DT-000623, DT-000811 and DT-000812. On Day 7 following the single-dose administration, mice were sacrificed, and RNA was collected from tissue for RNA extraction and quantitation of human PMP22 mRNA levels by quantitative RT-PCR. The average percent expression for each gene in the sciatic nerve and tibial nerve was calculated for each treatment and is shown in Table 38.
DT-000812 and DT-000945, an additional variant of DT-000623, were tested in C3-PMP22 mice. Groups of six C3-PMP22 mice each were treated with PBS or a single dose of 30 mg/kg of DT-000812 and DT-000945. One group of each treatment was sacrificed 14 days following the single-dose injection, and second groups of each treatment were sacrificed 28 days following the single-dose injection. RNA was collected from tissue for RNA extraction. Human PMP22 mRNA expression was measured by quantitative RT-PCR for both endpoints. Mouse MPZ, Pou3F1, and Sc5d mRNA levels were measured by quantitative RT-PCR for the 28-day endpoint. The average percent expression for each gene in the sciatic nerve, brachial plexus nerve, and tibial nerve was calculated for each treatment and time period and is shown in Tables 39 and 40.
To determine whether variations in siRNA nucleotide sequence and/or modified nucleotide pattern would yield compounds with improved properties such as potency and duration of action, further compounds targeting PMP22 were designed and tested. The structure of each compound is shown in Table 4.
Groups of four or five C3-PMP22 mice each were treated with PBS or a single dose of PBS or 30 mg/kg of conjugated siRNA compound. Seven days following injection, mice were sacrificed, and sciatic and brachial plexus nerves was collected for RNA extraction. Human PMP22 mRNA expression was measured by quantitative RT-PCR. The average percent expression for human PMP22 mRNA was calculated for each treatment and is shown in Table 41.
Groups of six C3-PMP22 mice each were treated with PBS or a single dose of PBS or 50 mg/kg of conjugated siRNA compound. Seven days following injection, mice were sacrificed, and sciatic and brachial plexus nerves was collected for RNA extraction. Human PMP22 mRNA expression was measured by quantitative RT-PCR. The average percent expression for human PMP22 mRNA was calculated for each treatment and is shown in Tables 42 through 49. For the compounds in Table 49, only the % human PMP22 remaining in the sciatic nerve is shown. Each table represents a different experiment.
Groups of six C3-PMP22 mice each were treated with a single dose of PBS, or 10 mg/kg or 30 mg/kg of conjugated siRNA compound (except for DT-000812 which was dosed only at 30 mg/kg). At Day 14 following injection, mice were sacrificed, and sciatic and brachial plexus nerve tissues were harvested for RNA extraction. Human PMP22 mRNA expression was measured by quantitative RT-PCR. The average percent expression for human PMP22 mRNA was calculated for each treatment and is shown in Tables 50 through 52. Each table represents a separate experiment.
C3-PMP22 mice are used as an experimental model of Charcot-Marie-Tooth disease type 1A (CMT1A). These transgenic mice express three to four copies of a wild-type human PMP22 gene, which leads to reduced numbers of myelinated fibers as early as three weeks of age. C3-PMP22 mice exhibit symptoms of neuromuscular impairment in the limbs similar to those observed in humans with CMT1A. Measurable functional endpoints in C3-PMP22 mice include, for example, motor nerve conduction velocity (MNCV), compound muscle action potential (CMAP), grip strength and beam walking.
The MNCV test is a non-invasive test that measures the velocity of a nerve signal. In this test, two electrodes are placed along a nerve, and the signal transduced between those electrodes is captured via a recording electrode placed at the neuromuscular junction. Defects in the myelin sheath in subjects with CMT1A cause a reduction in MNCV and a decrease in the amplitude of the transduced signal. These same findings are observed in C3-PMP22 mice.
CMAP is a quantitative measure of the amplitude of the electrical impulses that are transmitted to muscle. CMAP correlates with the number of muscle fibers that can be activated. In subjects with CMT1A, the CMAP of the nerve controlling contraction of the Anterior Tibialis muscle, a major muscle in the lower leg, correlates significantly with leg strength. These same findings are present in C3-PMP22 mice.
In the beam walking test, the dexterity of mice is observed as they walk along a horizontally suspended beam. Wild-type mice easily traverse the entire length of the beam. CMT1A mice, however, proceed more slowly and their paws may slip off the beam.
In the grip strength test, the mouse grasps a grid attached to a force transducer while an investigator gently pulls its tail. Grip strength is recorded as the force applied by the mouse in resisting the pulling motion. Relative to wild-type mice, grip strength of C3-PMP22 mice is reduced.
The efficacy of DT-000812 was evaluated in C3-PMP22 mice. Groups of six mice each were treated with PBS, weekly doses of 10 mg/kg DT-000812 (on Day 1 and weekly thereafter for a total of 11 doses), and monthly doses of 30 mg/kg DT-000812 (on Day 1, Day 28, and Day 56 for a total of 3 doses). Wild-type mice treated with PBS were used as a control (WT-PBS). Motor nerve conduction velocity (MNCV) and compound muscle action potential (CMAP) were determined just prior to treatment and at 4, 8, and 12 weeks to establish a baseline value for each endpoint. At 12 weeks, mice were sacrificed, and sciatic and brachial plexus nerves were harvested for RNA extraction. Human PMP22 mRNA expression in C3-PMP22 mice was measured by quantitative RT-PCR. Additionally, the expression of the top 500 dysregulated genes in wild-type mice relative to C3-PMP22 was evaluated by RNAseq. Peripheral nerves were dissected and prepared for morphometric analysis according to routine methods (for example, Jolivalt, et al., 2016, Curr. Protoc. Mouse Biol., 6:223-255). Cross sections of nerve were processed into resin blocks which were cut into 0.5- to 1.3-μm thick sections, stained with p-phenylenediamine, and viewed by light microscopy. Axon diameters and myelin thickness were measured using a software-assisted manual approach in ImageJ/FIJI.
The average percent expression for human PMP22 mRNA was calculated for each treatment and is shown in Table 53 and
The average MNCV per treatment group are shown in Table 54 and
The mean proportion of unmyelinated axons in each treatment group is shown in Table 57 and
In each table, WT-PBS indicates wild-type mice treated with PBS; all other data were obtained in C3-PMP22 mice (PBS, 10 mg/kg DT-000812, and 30 mg/kg DT-000812).
As illustrated by the above data, substantial improvements in multiple endpoints associated with CMT1A were observed.
Treatment of C3-PMP22 mice with DT-000812 resulted in a reduction in human PMP22 mRNA in both the sciatic and brachial plexus nerves (Table 53 and
The MNCV tests revealed an improvement in the efficiency of motor nerve conduction (Table 54 and
In wild-type mice, CMAP consisted of a strong electrical polarization signal, followed by a depolarization signal. In C3-PMP22 mice, both signals were muted and difficult to distinguish from background electrical impulses. In contrast, treatment with DT-000812 restored the shape and amplitude of CMAPs in C3-PMP22 mice (
In the beam walking test, wild-type mice easily traversed the entire length of the beam. In contrast, PBS-treated C3-PMP22 mice proceeded much more slowly, and their hind paws repeatedly slipped off the beam and on average required twice the amount of time to travel the same distance as wild-type mice. After twelve weeks of treatment of C3-PMP22 mice with DT-000812, the speed at which the mice traversed the beam was close to that of wild-type mice. Additionally, the number of slips relative to PBS-treated C3-PMP22 mice was reduced.
The grip strength of C3-PMP22 mice mice treated with PBS was markedly reduced relative to wild-type mice. Treatment with DT-000812 over a 12-week period increased forelimb grip strength to a level equivalent of wild-type mice. Furthermore, DT-000812 treatment over this same period led to increases in the mass of several peripheral muscles (quadricep and gastrocnemius) relative to untreated C3-PMP22 mice.
Measurement of nine genes essential for Schwann cell function illustrated that DT-000812 restored gene expression of these genes in the sciatic and brachial plexus nerves to the levels observed in wild-type mice. Additionally, RNAseq analysis revealed that the large majority of genes dysregulated in C3-PMP22 mice were restored toward wild-type levels of mRNA expression following treatment with DT-000812 at both the 10 mg/kg and 30 mg/kg doses.
Taken, these data demonstrate that inhibition of PMP22 with DT-000812 in C3-PMP22 mice, a model for CMT1A in human subjects, leads to substantial improvements in multiple phenotypes associated with CMT1A.
The efficacies DT-001246 and DT-001247 were evaluated, and compared to DT-000812, in C3-PMP22 mice. Groups of eight mice each were treated with PBS and a single dose of 30 mg/kg of each compound on Day 0 of the study. Motor nerve conduction velocity (MNCV) and compound muscle action potential (CMAP) were determined just prior to treatment (Baseline; Day -1) and at Day 27. At Day 28, mice were sacrificed, and sciatic and brachial plexus nerve tissues were harvested for RNA extraction. Human PMP22 mRNA expression was measured by quantitative RT-PCR.
The average percent expression for human PMP22 mRNA was calculated for each treatment and is shown in Table 59. MNCV and CMAP are shown in Table 60. The expression of several myelin-specific mouse mRNAs was also measured by quantitative RT-PCR. The average percent expression for each of these mRNAs was calculated and is shown in Table 61.
DT-000812, DT-001246, and DT-001247 were evaluated in a 60-day efficacy study in C3-PMP22 mice. Groups of eight mice each were treated with PBS and a single dose of 30 mg/kg of each compound on Day 0 of the study. Motor nerve conduction velocity (MNCV) and compound muscle action potential (CMAP) were determined just prior to treatment (Baseline; Day -1) and at Day 59. At Day 60, mice were sacrificed, and sciatic and brachial plexus nerve tissues were harvested for RNA extraction. Human PMP22 mRNA expression was measured by quantitative RT-PCR. The expression of several myelin-specific mouse mRNAs was also measured by quantitative RT-PCR.
The average percent expression for human PMP22 mRNA was calculated for each treatment and is shown in Table 62. MNCV and CMAP are shown in Table 63. The average percent expression for the myelin-specific mRNAs was calculated and is shown in Table 64.
The efficacies of DT-001250, DT-001251, DT-001252, and DT-001253 were evaluated, and compared to DT-000812, in C3-PMP22 mice. Groups of eight mice each were treated with PBS and a single dose of 30 mg/kg of each compound on Day 0 of the study. Motor nerve conduction velocity (MNCV) and compound muscle action potential (CMAP) were determined just prior to treatment (Baseline; Day -1) and at Day 27. At Day 28, mice were sacrificed, and sciatic and brachial plexus nerve tissues were harvested for RNA extraction. Human PMP22 mRNA expression was measured by quantitative RT-PCR.
The average percent expression for human PMP22 mRNA was calculated for each treatment and is shown in Table 65. MNCV and CMAP are shown in Table 66. The expression of several myelin-specific mouse mRNAs was also measured by quantitative RT-PCR. The average percent expression for each of these mRNAs was calculated and is shown in Table 67.
DT-000812, DT-001250, DT-001251, DT-001252, and DT-001253 were evaluated in a 60-day efficacy study in C3-PMP22 mice. Groups of eight mice each were treated with PBS and a single dose of 30 mg/kg of each compound on Day 0 of the study. Motor nerve conduction velocity (MNCV) and compound muscle action potential (CMAP) were determined just prior to treatment (Baseline; Day -1), at Day 28 and at Day 59. At Day 60, mice were sacrificed, and sciatic and brachial plexus nerve tissues were harvested for RNA extraction. Human PMP22 mRNA expression was measured by quantitative RT-PCR. The expression of several myelin-specific mouse mRNAs was also measured by quantitative RT-PCR.
The average percent expression for human PMP22 mRNA was calculated for each treatment and is shown in Table 68. MNCV and CMAP are shown in Table 69. The average percent expression for the myelin-specific mRNAs was calculated and is shown in Table 70.
The efficacies of DT-001254, DT-001255, and DT-001257 were evaluated in C3-PMP22 mice. DT-000812 was included in the study. Groups of eight mice each were treated with PBS and a single dose of 30 mg/kg of each compound on Day 0 of the study. Wild-type mice treated with PBS were used as a control (WT-PBS). Motor nerve conduction velocity (MNCV) and compound muscle action potential (CMAP) were determined just prior to treatment (Baseline; Day -1) and at Day 27. At Day 28, mice were sacrificed, and sciatic and brachial plexus nerve tissues were harvested for RNA extraction. Human PMP22 mRNA expression was measured by quantitative RT-PCR. The expression of several myelin-specific mouse mRNAs was also measured by quantitative RT-PCR.
The average percent expression for human PMP22 mRNA was calculated for each treatment and is shown in Table 71. MNCV and CMAP are shown in Table 72. The average percent expression for myelin-specific mouse mRNAs was calculated and is shown in Table 73.
In each table, WT-PBS indicates wild-type mice treated with PBS; all other data were obtained in C3-PMP22 mice.
DT-000812, DT-001254, DT-001255, and DT-001257 were evaluated in a 60-day efficacy study in C3-PMP22 mice. Groups of eight mice each were treated with PBS and a single dose of 30 mg/kg of each compound on Day 0 of the study. Wild-type mice treated with PBS were used as a control (WT-PBS). Motor nerve conduction velocity (MNCV) and compound muscle action potential (CMAP) were determined just prior to treatment (Baseline; Day −1), at Day 28 and at Day 59. At Day 60, mice were sacrificed, and sciatic and brachial plexus nerve tissues were harvested for RNA extraction. Human PMP22 mRNA expression was measured by quantitative RT-PCR. The expression of several myelin-specific mouse mRNAs was also measured by quantitative RT-PCR.
The average percent expression for human PMP22 mRNA was calculated for each treatment and is shown in Table 74. MNCV and CMAP are shown in Table 75. The average percent expression for the myelin-specific mRNAs was calculated and is shown in Table 76.
In each table, WT-PBS indicates wild-type mice treated with PBS; all other data were obtained in C3-PMP22 mice.
The efficacy of DT-001263 was evaluated and compared to DT-000812, in C3-PMP22 mice. Groups of eight mice each were treated with PBS and a single dose of 30 mg/kg of each compound on Day 0 of the study. Wild-type mice treated with PBS were used as a control (WT-PBS). Motor nerve conduction velocity (MNCV) and compound muscle action potential (CMAP) were determined just prior to treatment (Baseline; Day -1) and at Day 27. At Day 28, mice were sacrificed, and sciatic and brachial plexus nerve tissues were harvested for RNA extraction. Human PMP22 mRNA expression was measured by quantitative RT-PCR.
The average percent expression for human PMP22 mRNA was calculated for each treatment and is shown in Table 77. MNCV and CMAP are shown in Table 78. The expression of mouse MPZ mRNA was also measured by quantitative RT-PCR. The average percent expression for each of these mRNAs was calculated and is shown in Table 79.
In each table, WT-PBS indicates wild-type mice treated with PBS; all other data were obtained in C3-PMP22 mice.
12-week Efficacy Studies: DT-001252, DT-001253, and DT-001257
DT-001252, DT-001253, and DT-001257 were each evaluated in separate 12-week efficacy studies in C3-PMP22 mice. Each study also included treatment with DT-000812 at 30 mg/kg. Groups of eight mice each were treated with PBS, or monthly doses of 3 mg/kg, 10 mg/kg, or 30 mg/kg siRNA compound on Day 0, Day 28, and Day 56, for a total of 3 doses. Wild-type mice treated with PBS were used as a control (WT-PBS). Motor nerve conduction velocity (MNCV), compound muscle action potential (CMAP), grip strength and beam walking ability were determined just prior to treatment to establish a baseline value and at 4, 8, and 12 weeks of treatment. At 12 weeks, mice were sacrificed, and sciatic and brachial plexus nerves were harvested for RNA extraction. Human PMP22 mRNA expression in C3-PMP22 mice was measured by quantitative RT-PCR. The expression of several myelin-specific mouse mRNAs was also measured by quantitative RT-PCR. Peripheral nerves were dissected and prepared for morphometric analysis according to routine methods (for example, Jolivalt, et al., 2016, Curr. Protoc. Mouse Biol., 6:223-255). Cross sections of nerve were processed into resin blocks which were cut into 0.5- to 1.3-μm thick sections, stained with p-phenylenediamine, and viewed by light microscopy. Axon diameters and myelin thickness were measured using a software-assisted manual approach in ImageJ/FIJI.
The average percent expression for human PMP22 mRNA was calculated for each treatment and is shown in Table 80. The average percent expression for myelin-specific mRNAs was calculated and is shown in Table 85.
The average MNCV per treatment group at each time point is shown in Table 81. In the experiment testing DT-001252, errors in measurement of the traces resulted in variable MNCV data at the baseline, 4-week and 8-week timepoints, thus these data are not presented.
The average CMAP per treatment group at each time point is shown in Table 82. Grip strength and beam walking ability were measured at 4, 8, and 12 weeks and are shown in Table 82.
The mean percentage of unmyelinated axons in each treatment group is shown in Table 83.
In each table, WT-PBS indicates wild-type mice treated with PBS; all other data were obtained in C3-PMP22 mice.
As illustrated by the above data, substantial improvements in multiple endpoints associated with CMT1A were observed.
Treatment of C3-PMP22 mice with each conjugated PMP22 siRNA tested resulted in a reduction in human PMP22 mRNA expression compared to PBS-treated C3-PMP22 mice in both the sciatic and brachial plexus nerves (Table 80).
The MNCV tests revealed an improvement in the efficiency of motor nerve conduction at 12 weeks (Table 81). Additionally, each conjugated PMP22 siRNA tested improved compound muscle action potential at each time point (Table 82). The improvement in CMAP following treatment with DT-001252 is further illustrated in
The grip strength of C3-PMP22 mice mice treated with PBS was markedly reduced relative to wild-type mice. Treatment with the conjugated PMP22 siRNAs increased grip strength (Table 83). Furthermore, increases in the masses of several peripheral muscles (quadricep, tibialis anterior and gastrocnemius) were increased relative to untreated C3-PMP22 mice. In the beam walking test, wild-type mice easily traversed the entire length of the beam. In contrast, PBS-treated C3-PMP22 mice proceeded much more slowly, and their hind paws repeatedly slipped off the beam and on average required additional time to travel the same distance as wild-type mice. After treatment with the conjugated PMP22 siRNAs, the speed at which C3-PMP22 mice traversed the beam was closer to that of wild-type mice (Table 85). Additionally, the number of slips relative to PBS-treated C3-PMP22 mice was reduced (Table 84).
Measurement of myelin-specific genes essential for Schwann cell function illustrated that treatment with the conjugated PMP22 siRNAs restored gene expression of these genes in the sciatic and brachial plexus nerves to the levels observed in wild-type mice (Table 83).
Taken, these data demonstrate that inhibition of PMP22 with conjugated PMP22 siRNAs, in an experimental model for CMT1A, leads to substantial improvements in multiple phenotypes associated with CMT1A.
The efficacy of DT-001252 was further evaluated by measuring myelination of the femoral motor nerve. Peripheral nerves were dissected and prepared for morphometric analysis according to routine methods (for example, Jolivalt, et al., 2016, Curr. Protoc. Mouse Biol., 6:223-255). Cross sections of nerve were processed into resin blocks which were cut into 0.5- to 1.3-μm thick sections, stained with p-phenylenediamine, and viewed by light microscopy. Axon diameters and myelin thickness were measured using a software-assisted manual approach in ImageJ/FIJI. Histological analysis revealed that, whereas unmyelinated axons were common in femoral motor nerve sections from C3-PMP22 mice, each DT-001252 treatment group exhibited substantially lower numbers of large unmyelinated axons (Table 87,
The effect of treatment with DT-001252 on serum Neurofilament light (NfL) was also evaluated. NfL is a marker of neuronal damage and is elevated in subjects with CMT1A. Serum NfL at 12 weeks was measured using a NFL-light Advantage assay kit (Quanterix). The mean NfL for each treatment group is shown in Table 88 (n=7 for PBS-treated C3-PMP22 mice due to exclusion of one outlier individual data point; n=8 for all other groups). As shown in Table 88, treatment with each dose of DT-001252 normalized serum NfL.
Additional compounds were designed to evaluate the effects of chemical modifications on the potency of certain conjugated PMP22 siRNAs related to unconjugated compounds identified as “hits” and shown in Table 19. These derivatives comprise the identical nucleotide sequences as their respective parent compounds but have variations in nucleotide modifications. DT-001842 and DT-001843 are derivatives of DT-000901; DT-001844 and DT-001845 are derivatives of DT-000847; DT-001846 and DT-001847 are derivatives of DT-000849; DT-001848 and DT-001849 are derivatives of DT-000855; DT-001858, DT-001859, and DT-001860 are derivatives of DT-000414. Groups of five C3-PMP22 mice each were treated with a single dose of PBS, or 10 mg/kg or 30 mg/kg of conjugated siRNA compound. DT-001252 was included in each study as a benchmark compound. At Day 14 following injection, mice were sacrificed, and sciatic and brachial plexus nerve tissues were harvested for RNA extraction. Human PMP22 mRNA and mouse MPZ mRNA were measured by quantitative RT-PCR. The average percent expression for each mRNA was calculated for each treatment and is shown in Tables 89 through 94. As illustrated in the tables below, derivatives of DT-001252 exhibited potency comparable to that of DT-001252.
Comparison of Activity of Structurally Related Conjugated PMP22 siRNAs
As illustrated herein, certain conjugated PMP22 siRNAs exhibited potent reduction of hPMP22 in the C3-PMP22 mouse model. One such group of related siRNAs is listed in Table 95. Each of these siRNAs has the sense strand of SEQ ID NO: 1015 or SEQ ID NO: 1018 (which differ by a single nucleobase), the antisense strand of SEQ ID NO: 1144 and the DTx-01-08 motif conjugated to the 3′ end of the sense strand through a C7 linker as described herein. As each antisense strand of each siRNA has the nucleotide sequence of SEQ ID NO: 1144, each siRNA targets nucleotides 213 to 233 of the human PMP22 mRNA. Variations were introduced in the number, nature, and placement of chemical modifications, as shown in Table 95. Each 0o hPMP22 shown in Table 95 is from an experiment described herein and is reproduced below for comparison. While each of the conjugated PMP22 siRNAs in Table 95 exhibits potent reduction of the hPMP22 mRNA, certain analogs including but not limited to DT-001252 and DT-001253 are notable for their duration of action.
MGFUMAFUMCFSAMSUF-C7OH-[DTx-
MAFCMAFGMGFAMGFGMSAMSGM-OH-3′
MGFUMAFUMCFSAMSUF-C7OH-[DTx-
MAFCMAFGMGFAMGFGMSAMSGM-OH-3′
FAMGFUMAFUMCMSAMSUM-C7OH-
MAFCMAFGMGFAMGFGMSAMSGM-OH-3′
MAMGMUMAMUMCMSAMSUM-C7OH-
MAMGMUMAMUMCMAMUM-C7OH-
MGFUMAFUMCMSAMSUM-C7OH-[DTx-
MAMGMUMAMUMCMAMSUM-C7OH-
MAMGMUMAMUMCMSAMSUM-C7OH-
This application is a continuation of International Patent Application No. PCT/US2022/080012, which claims the benefit of U.S. Provisional Application No. 63/280,773 filed Nov. 18, 2021, the contents of each of which are hereby incorporated herein in their entirety and for all purposes.
This invention was made with government support under grant number 1R43NS119090-01A1 awarded by the National Institutes of Health. The government has certain rights in the invention.
| Number | Date | Country | |
|---|---|---|---|
| 63280773 | Nov 2021 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/US2022/080012 | Nov 2022 | WO |
| Child | 18660600 | US |
