Substituted RIG-I agonists: compositions and methods thereof

SEQUENCE LISTING

The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Jun. 22, 2021, is named 24588USPCT-SEQLIST-22JUN2021.txt and is 2320 Kb in size.

BACKGROUND OF THE INVENTION

Retinoic acid-inducible gene 1 (RIG-I) plays a crucial role in promoting the release of type I and type III interferons to fortify host anti-viral immunity (Wu et al., 2015, Virology 482: 181-188). The RIG-I pathway has been identified as an important innate immune sensing pathway of cytosolic 5′-triphosphate (5′ppp) dsRNA that drives the production of a variety of cytokines, including interferon β. In response to viral 5′ppp cytosolic double stranded RNA, RIG-I undergoes activation and recruits mitochondrial antiviral signaling protein (MAVS). Moreover, transcriptome analysis reveals a RIG-I-related signature covering the canonical pathway categories “IFN signaling”, “activation of IRFs by cytosolic pattern recognition receptors (PRRs)”, “TNFR2 signaling” and “antigen presentation” indicating that RIG-I bridges the innate and adaptive immune system (Goulet et al., 2013, PLoS Pathogens). Intriguingly, RIG-I-induced immunogenic tumor cell death triggers adaptive immunity, engaging dendritic cells and T-cells to kill tumors in vivo providing a second innate/adaptive immune system loop (Duewell et al., Cell death and differentiation 2014: 21(12) 1825-1837). Of note, RIG-I-induced apoptosis is restricted to tumor cells only (Duewell et al., 2014, 21(12) 1825-1837).

Recent studies identified key structural features for optimal RIG-I ligands. Short length, double-strandedness, 5′-triphosphorylation and blunt end base pairing characterize the prototypic RNA-based RIG-I agonist (Schlee et al., 2009, Immunity 31:25-34; Pichlmair et al., Science 2006, 314: 997-1001; Schlee, Immunobiology, 2013; 218(11), 1322-1335). Moreover, circular structures (Chen et al. Mol Cell 67 (2), 228-238.e5. 2017) and bent/KINK RNAs (Lee et al., 2016, Nucleic Acids Res, 44(17), 8407-8416) constitute another recently identified group of RIG-I ligands that do not require a triphosphate moiety. Nabet et al. (Cell 2017 Jul. 13; 170(2):352-366.e13. doi: 10.1016/j.cell.2017.06.031) reported that an unshielded endogenous RNA can activate RIG-I in tumor cells promoting aggressive features of cancer. Recent findings indicate also that endogenous small non-coding RNAs leaking to the cytoplasm can activate RIG-I during ionizing radiation therapy (Ranoa et al., Oncotarget 2016; 7(18), 26496-26515). In addition, a hetero-trimeric complex of RIG-I/RNA polymerase III/serine-arginine-rich splicing factor 1 facilitates RIG-I activation in response to delocalized, cytosolic DNA via a 5′ppp RNA intermediate (Ablasser et al., Nat Immunol 2009; 10(10), 1065-1072; Xue et al., PLoS One 2015; 10920, E0115354).

Structural and functional analysis of RIG-I reveals that single amino acids and a lysine-rich patch located at the C-terminal domain (CTD) of RIG-I sense the structural properties of RNAs (Wang et al., Nat Struc Mol Biol 2010; 17(7), 781-787). Remarkably, a typical eukaryotic 2′-O-methylation pattern along with 7-methyl guanosine capping of the 5′-triphosphate group of RNAs prevent binding to RIG-I, thus distinguishing host from pathogenic non-self RNA. Specifically, modifications at the 5′ end decrease RNA affinity and production of pro-inflammatory cytokines (Schuberth-Wagner et al., 2015, Immunity 43, 41-51; Devarkar et al., PNAS 2016; 113(3), 596-601). Modified RNAs containing modified nucleotides m6A, Ψ, mΨ, 2FdU, 2FdC, 5mC, 5moC, and 5hmC appear to lack stimulatory activity (Durbin et al., mBio 7(5):e00833-16. doi: 10.1128/mBio.00833-16). Moreover, illegitimate RIG-I activation by endogenous RNA is controlled by fast ATPase turnover which leads to dissociation of the RIG-I/RNA complex (Louber et al., BMC Biology 2015; 13:54). Interestingly, RIG-I mutations that correlate with a decreased ATPase activity appear to be constitutively active potentially due to signals from host RNA (Fitzgerald et al., 2017, Nucleic Acids Res, 45(3), 1442-1454).

Improvement in the understanding of how RIG-I recognizes RNA and utilizes ATP has resulted from structural studies of RIG-I truncations in mouse, human, and duck (Civril et al., 2011, EMBO Rep 12:1127-1134; Jiang et al., 2011, Nature 479:423-427; Kowalinski et al., 2011, Cell 147:423-435; Luo et al., 2011, Cell 147:409-422). However, in the only RIG-I structure with caspase activation and recruitment domains (CARD) present, the protein was in an inactive, apo-state, and lacked the CTD. This leads to questions regarding the role of both RNA and ATP in the innate immune response derived from RIG-I activation, and the relative positions of the CTD and CARDs in the active RIG-I conformation. Interestingly, the RIG-I CTD capped the 5′ end of the RNA, regardless of the length of the bound duplex. The preference for the end of the duplex RNA in these structures was also independent of a 5′ppp. Recent publications, including those mentioned herein, have demonstrated that activation of the innate immune system through RIG-I leads to potent anti-tumor immunity. This new immunotherapeutic mechanism may be synergized with checkpoint inhibitors such as an anti-PD1 antibody to achieve greater anti-tumor responses. See also Elion et al., 2018, Cancer Research (DOI: 10.1158/0008-5472.CAN-18-0730).

Hence, there still remains a need for understanding RIG-I functionality and establishing RIG-I selectivity, boosting or abrogating RIG-I-driven immunity.

SUMMARY OF THE INVENTION

The present invention provides a composition comprising a nucleic acid capable of activating RIG-I. This invention provides compounds, and uses thereof, wherein said compounds comprise a short single strand oligonucleotide that optionally forms an oligonucleotide hairpin or stem loop molecule with self complementary base pairing of less than 12 base pairs that bind to RIG-I and activate the RIG-I pathway. An embodiment of this invention is realized when the short single strand oligonucleotide does not form an oligonucleotide hairpin or stem loop molecule.

An embodiment of this invention is realized when the short single strand oligonucleotide forms an oligonucleotide hairpin or stem loop molecule. An embodiment of this invention is realized when the short oligonucleotide hairpin molecule optionally comprises a 5′-terminus group selected from OH, diphosphate (pp) group and triphosphate (ppp) group. Another embodiment of this invention is realized when the short oligonucleotide hairpin molecule optionally comprises a 5′-terminus selected from functional groups that mimic the interactions of the triphosphate and diphosphate groups, sometimes referred to as 5′ terminus pp mimic or ppp mimic. An embodiment of this invention is realized when in the absence of a 5′-terminus phosphate group the short oligonucleotide molecule comprises a 5′-terminus that is a 5′-OH. Another embodiment of this invention is realized when the short oligonucleotide hairpin molecule comprises a 5′ terminus pp or 5′ terminus pp mimic. Another embodiment of this invention is realized when the short oligonucleotide hairpin molecule comprises a 5′-terminus ppp, or a 5′-terminus ppp mimic.

An embodiment of this invention is realized when the short oligonucleotide molecule comprises a 5′-terminus ppp. Still another sub-embodiment of this invention is realized when the short oligonucleotide molecule comprises a 5′-terminus ppp mimic. In yet another embodiment the short oligonucleotide hairpin molecule comprises a 5′-terminus group selected from diphosphate and triphosphate, a self complementary base pairing region of less than 12 base pairs, and a blunt end. In another embodiment, the short oligonucleotide hairpin molecule comprises a 5′-terminus mimic group selected from diphosphate and triphosphate, a self complementary base pairing region of less than 12 base pairs, and a blunt end. In another embodiment, the short oligonucleotide hairpin molecule comprises a 5′-terminus mimic group selected from diphosphate and triphosphate, a self complementary base pairing region of less than 12 base pairs, a blunt end, and a non-base pairing stem loop. A subembodiment of this aspect of the invention is realized when the looped end is a non-base pairing tetraloop consisting of four nucleotides. Another subembodiment of this aspect of the invention is realized when a blunt end is at the 5′terminus end.

Still in another embodiment, the invention is directed to short oligonucleotide hairpin or stem loop molecules having a self complementary base pairing region of less than 12 base pairs, a blunt end, and a non-base pairing tetraloop, further comprising a modified phosphodiester backbone. A sub-embodiment of this aspect of the invention is realized when the modified phosphodiester backbone comprise at least one modified phosphate group comprising a phosphorothioate.

In another embodiment of this aspect of the invention, the short oligonucleotide hairpin molecules comprise at least one modified nucleotide base (nucleobase).

In yet another embodiment, disclosed is a method for inducing type I interferon production in a cell, or a method for treating a disease or disorder in a subject in need thereof by inducing type I interferon production in a cell of the subject. In another embodiment, use of one or more short oligonucleotide molecules in the potential treatment or prevention of RIG-I pathway-associated diseases or disorders is disclosed, wherein the molecule comprises a self-complementary base pairing region of less than 12 base pairs, a blunt end, and non base pairing tetraloop. In a sub-embodiment of this aspect of the invention, the disease or disorder is selected from the group consisting of a bacterial infection, a viral infection, a parasitic infection, cancer, an autoimmune disease, an inflammatory disorder, and a respiratory disorder.

In still another embodiment, compositions comprising one or more of short oligonucleotide molecules are disclosed. In another embodiment, a pharmaceutical composition comprising one or more of the short oligonucleotide hairpin molecules capable of inducing interferon production and a pharmaceutically acceptable carrier, wherein the molecule comprises a self complementary base pairing region of less than 12 base pairs, a blunt end and a non base pairing tetraloop. In a sub-embodiment of this aspect of the invention, the 5′-terminus of the short oligonucleotide hairpin molecules comprise at least one of the group consisting of 5′-OH, 5′-triphosphate, 5′-diphosphate, 5′-diphosphate mimic, and 5′-triphosphate mimic. In a sub-embodiment of this aspect of the invention, the pharmaceutical composition further comprises at least one agent selected from an immunostimulatory agent, an antigen, an anti-viral agent, an anti-bacterial agent, an anti-tumor agent, retinoic acid, IFN-α, and IFN-β.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to novel oligonucleotides and analogues thereof and their use to bind and activate the RIG-I protein.

Embodiments of the disclosure include compounds of general Formula Ia and Formula I as well as synthesis and isolation of compounds of general Formula Ia and Formula I. Uses of compounds of general Formula Ia and Formula I are also disclosed. Other embodiments, aspects and features of the present disclosure are either further described in or will be apparent from the ensuing description, examples and appended claims.

The term “oligonucleotide” in the context of the present application encompasses compounds comprising a plurality, e.g., at least 4 nucleotide or nucleotide analogue building blocks, preferably, 6-100, e.g., 19-40, 19-30, and 19-25 building blocks. The nucleotide or nucleotide analogue building blocks may comprise nucleoside or nucleoside analogue subunits connected by inter-subunit linkages. The nucleoside subunits include deoxyribonucleoside subunits, ribonucleoside subunits and/or analogues thereof, particularly sugar- and/or nucleobase-modified nucleoside analogues. Further, the oligonucleotides may comprise non-nucleotidic building blocks and/or further terminal and/or side-chain modifications.

Examples of sugar-modified subunits are realized when R₁, R₂R₃, and/or R₄of a ribonucleoside subunit is replaced, for example, by a group as defined below. In other sugar-modified subunits, the ribose may be substituted, e.g., by another sugar, for example a pentose such as arabinose. This sugar modification may be combined with 2′-OH modifications as described above, such as in 2′-fluoroarabinonucleoside subunits. Still other sugar-modified subunits include locked nucleosides (LNA). A “locked nucleoside” is a nucleoside in which the robose moiety is modified with an extra bridge connecting a 2′ oxygen and a 4′ carbon. Still other nucleobase-modified nucleosidic building blocks, a non-standard, e.g., non-naturally occurring nucleobase, is used instead of a standard nucleobase. Examples of non-standard or modified nucleobases are uracils or cytosines modified at the 5-position, e.g., 5-(2-amino)propyl uracil or 5-bromouracil; hypoxanthine; 2,6-diaminopurine; adenines or guanines modified at the 8-position, e.g. 8-bromoguanine; deazanucleosides, e.g. 7-deazaguanine or 7-deazaadenine; or O- and N-alkylated nucleobases, e.g. N⁶-methyladenine, or N⁶,N⁶-dimethyladenine. Further suitable nucleobase analogues may be selected from universal nucleobase analogues such as 5-nitroindole or inosine.

As used herein, the term “backbone” refers to the linkage between the subunits of the oligonucleotide. A “modified phosphodiester backbone” occurs when the inter-subunit linkage between subunits is be a phosphodiester linkage or a modified linkage, e.g. a phosphorothioate, phosphorodithioate, methylphosphonate, phosphoramidate, boranophosphate, and the like or another modified linkage described herein or known to a skilled person in the art.

As used herein, a “single stranded oligonucleotide” refers to an oligonucleotide in which the bases are not complementary to each other. A “hairpin” oligonucleotide structure refers to a single stranded oligonucleotide, wherein a portion of the oligonucleotide chain loops around and self complementary base pairing results in a portion of the oligonucleotide which is effectively double stranded.

As used herein, a “double stranded oligonucleotide,” refers to an oligonucleotide of two strands, which is formed from complementary base pairsA double-stranded oligonucleotide as used herein can have terminal overhangs on either end or both ends. An oligonucleotide section is fully double-stranded when the two stretches of nucleic acid forming the section have the same length and have sequences which are 100% complementary to each other. As established in the art, two nucleotides are said to be complementary to each other if they can form a base pair, either a Waston-Crick base pair (A-U, G-C) or a wobble base pair (U-G, U-A, I-A, I-U, I-C).

As used herein, the term “blunt end” as used in a double stranded oligonucleotide refers to the 3′ and 5′ ends of the oligonucleotide of the invention, wherein the oligonucleoide terminates in a base pair. Oligonucleotides which do not have a blunt end are referred to as having an overhang, wherein one strand has a longer oligonucleotide chain than the other strand. A single stranded oligonucleotide may have a single blunt end or an overhang.

The oligonucleotide may be selected from deoxyribonucleotides, ribonucleotides and oligonucleotide analogues. The deoxyribonucleotides, ribonucleotides and/or oligonucleotide analogues may be chemically modified at the nucleoside and/or ribose subunit of the analogue, deoxyribonucleotide, ribonucleotide and/or oligonucleotide. Analogues of desoxyribonucleotides or ribonucleotides may comprise at least one desoxyribonucleoside or ribonucleoside subunit and at least one modified nucleosidic subunit and/or at least one modified inter-subunit linkage, e.g. as described above. Oligonucleotide analogues may also consist in their entirety of modified nucleosidic subunits.

The oligonucleotides of the present invention comprise short single strand oligonucleotides that optionally may fold to form a hairpin consisting of self complementary base pairings and an unpaired tetraloop. The self complementary base pairing hairpin structure is formed when intra-molecular base pairing forms a double helix that ends in an unpaired tetraloop opposite the 5′ terminus end of the single strand. An embodiment of this aspect of the invention is realized when the self complementary base pairing is less than 12 base pairs and the unpaired loop is a tetraloop of four nucleotides. The hairpin molecules may be blunt ended or comprise at least one overhang, e.g., a 5′- or 3′-overhang. The hairpin molecules may optionally comprise a 5′ OH group, 5′-terminus phosphate group or 5′-terminus phosphate mimic group.

The oligonucleotides of the present invention comprise single strands of a length of at least 24 nucleotides. In another embodiment, the single strand has a length of 24 nucleotides.

The oligonucleotide may comprise further terminal and/or side-chain modifications, e.g., cell specific targeting entities covalently attached thereto. Those entities may promote cellular or cell-specific uptake and include, for example lipids, small molecules, cholesterol, folate, vitamins, hormones, peptides, oligosaccharides and analogues thereof. Targeting entities may, e.g., be attached to modified nucleobases or non-nucleotidic building blocks by methods known to the skilled person.

Still in another embodiment, modifications establish and/or enhance the selectivity of the oligonucleotide towards a given target. In a particularly preferred embodiment the RIG-I selectivity of the oligonucleotide is enhanced. Methods to determine the RIG-I selectivity of a given oligonucleotide are described herein in detail (cf. Examples) and/or are known to the person skilled in the art.

Yet in another embodiment, the chemical modifications maintain or enhance the chemical stability of the oligonucleotide. A person skilled in the art knows methods for determining the chemical stability of a given oligonucleotide. Such methods are also described, e.g., in the Examples.

Chemical modifications of the oligonucleotide include those to the ribonucleoside subunit, phosphodiester backbone, and nucleobase. Non-limiting examples of such modifications are independently selected from the group comprising halogenation, (e.g., 2′-F-halogenation), 2′-O-alkylation (e.g., 2′-O-methylation), and/or phosphorothioate modifications of internucleotide linkages. Particularly, 2′-F-halogenation and phosphorothioate modifications may increase the stability of the oligonucleotide, while 2′-O-methylation may establish or increase RIG-I selectivity of the oligonucleotide. 2′-O-methylations may also be able to modify the immunogenicity of RNA.

The identification patterns of a given oligonucleotide depend on the sequence and the length of an oligonucleotide and can be determined for each given oligonucleotide. A person skilled in the art is well aware how to carry out this determination. As explained herein, such methods for determining RIG-I selectivity and/or stability of a given oligonucleotide are described in detail in the present application.

The present invention provides modified short oligonucleotide molecules of general Formula Ia:

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or pharmaceutically acceptable salts thereof, wherein:

each X is independently selected from the group consisting of O, S, —CH₂—, and —NH—;

each Y is independently selected from the group consisting of —OH, —SH, and

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each V is independently O, or S;

K is H, or

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each W is independently N, —CH—, or —C(F)—;

each Z is independently N, or —CH—;

each R₁is independently selected from the group consisting of H, halogen, —OH, —OC_1-6alkyl, C_1-6alkyl, —OC_1-3haloalkyl, —OC_2-6alkenyl, —OC_2-6alkynyl, —NH₂, and —OCH₂C_5-6heteroarylCH₂R_y, said alkyl, alkenyl, alkynyl and heteroaryl optionally substituted with 1 to 3 groups of R_b;

each R₂is independently selected from the group consisting of H, halogen, —OH, —OC_1-6alkyl, C_1-6alkyl, —OC_1-3haloalkyl;

or one or more adjacent R₁and R₂can combine to form ═CH₂;

each R₃is independently H, halogen, C_1-6alkyl, or C_2-6alkynyl;

or R₁and R₃on a sugar moiety can be linked to form —CH₂O, provided R₂is hydrogen;

R₄is selected from the group consisting of —CH₂OH,

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R₅is selected from the group consisting of H, C_1-6alkyl, —(CH₂CH₂O)_pCH₂CH₃, and —CH₂CH(OH)CH₂(O(CH₂)₂)_nCH₂NHC(O)R_z;

each R₆is independently selected from H and NH₂;

R_bis selected from C_1-6alkyl, —OC_1-6alkyl, halo, OH, CN, NO, NH₂, NHC_1-6alkyl, N(C_1-6alkyl)₂, SO₃H, SO₄, PO₄, C_1-3haloalkyl, C_1-3haloalkoxy, carboxyl, oxo, and S(O)_galkyl;

R is H or —P(O)(OH)_2′;

Q is independently selected from the group consisting of O, CH₂, and NH;

R_sis NH₂or —C(O)—;

R_xis hydrogen or —C≡CH;

R_yis —CH₂(O(CH₂)₂)_n(CH₂)_kNHC(O)—R_z, or —(CH₂OCH₂)_nCH₂C(O)N((CH₂)_kNHC(O)R_z)₂;

R_zis cholesterol;

g is 1, 2, or 3;

k and n are independently selected from 1, 2, 3, 4, 5, and 6;

p is an integer from 1 to 50.

An embodiment of the invention of Formula Ia is represented by Formula I:

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each V is independently O, or S;

K is H, or

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R₅is selected from the group consisting of H, C_1-6alkyl, —(CH₂CH₂O)_pCH₂CH₃, and —CH₂CH(OH)CH₂(O(CH₂)₂)_nCH₂NHC(O)R_z;

each R₆is independently selected from H and NH₂;

R_bis selected from C_1-6alkyl, —OC_1-6alkyl, halo, OH, CN, NO, NH₂, NHC_1-6alkyl, N(C_1-6alkyl)₂, SO₃H, SO₄, PO₄, C_1-3haloalkyl, C_1-3haloalkoxy, carboxyl, oxo, and S(O)_galkyl;

R is H or —P(O)(OH)_2′;

Q is independently selected from the group consisting of O, CH₂, and NH; R_yis —CH₂(O(CH₂)₂)_n(CH₂)_kNHC(O)—R_z, or —(CH₂OCH₂)_nCH₂C(O)N((CH₂)_kNHC(O)R_z)₂;

R_zis cholesterol;

g is 1, 2, or 3;

k and n are independently selected from 1, 2, 3, 4, 5, and 6;

p is an integer from 1 to 50.

For purposes of this invention, the compound of Formula Ia and I comprises 24 nucleotides selected from the group consisting of 1, 2, 3, 4, 5, and 6,

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wherein R₁, R₂, R₃, R₄, R₆, R_x, R_s, X, V, Y, K, Z, and W are as described herein. Adenine is present at positions 5, 9, 12, 17, 18, 19 and 22. Uracil is present at positions 3, 6, 7, 8, 16 and 20. When the hairpin is present, and reading from left to right, up to 10 self complementary base pairs are possible.

The compounds of the invention also include adenine derivative, inosine, or derivatives thereof, as depicted below:

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An embodiment of the invention of Formula Ia and I is realized when it is read from left to right beginning with guanine (G), which is at the 5′ end, and ending with cytosine (C) at the 3′ end.

Another embodiment of the invention is compounds of formula (I), or a pharmaceutically acceptable salt thereof, wherein any X is independently selected from O and S, any Y is independently selected from OH and SH, any V is independently selected from O or S; and any K is independently selected from H and OH

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In another embodiment, any R₁is independently selected from the group consisting of H, halogen, —OH, —OMe, Me, —OCF₃, O(CH₂)₂OCH₃, NH₂, —OCH₂C≡CH, any R₂is independently selected from the group consisting of H, halogen, —OH, —OMe, Me, and —OCF₃.

In another embodiment, any X is independently selected from O and S, any Y is independently selected from OH and SH, any V is independently selected from O or S; K is H, any W and Z within a single nine membered bicycle is independently selected from W═N and Z═—CH—; W═—CH— and Z═N and W═—CH— and Z is CH, any R₁is selected from the group consisting of H, halogen, —OH, —OMe, Me, —OCF₃, —O(CH₂)₂OCH₃, —NH₂, —OCH₂C≡CH, any R₂is independently selected from the group consisting of H, halogen, —OH, —OMe, -Me, and —OCF₃, and any R₃is independently selected from the group consisting of H, halogen, C_1-6alkyl, and —C≡CH.

An embodiment of the invention of Formula Ia and I is realized when any X is independently selected from O, S, and —CH₂—. A subembodiment of this aspect of the invention is realized when any X is independently selected from O and S. A subembodiment of this aspect of the invention is realized when each X is O. A subembodiment of this aspect of the invention is realized when each X is S. A subembodiment of this aspect of the invention is realized when each X is —CH₂—.

Another embodiment of the invention of Formula Ia and I is realized when any Y is independently selected from OH and SH A subembodiment of this aspect of the invention is realized when each Y is SH. A subembodiment of this aspect of the invention is realized when each Y is OH.

An embodiment of this aspect of the invention is realized when one to three Y is

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A further subembodiment of this aspect of the invention is realized when the heteroaryl is optionally substituted triazolyl, said triazolyl optionally substituted with 1 to 3 groups of R_b. Another embodiment of this aspect of the invention is realized when R_yis —CH₂(O(CH₂)₂)_n(CH₂)_kNHC(O)—R_z. Still another subembodiment of this aspect of the invention is realized when R_yis —(CH₂OCH₂)_nCH₂C(O)N((CH₂)_kNHC(O)R_z)₂. Another subembodiment of this aspect of the invention is realized when Y is

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selected from

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Another embodiment of the invention of Formula Ia and I is realized when V is O. Another subembodiment of the invention of Formula Ia and Formula I is realized when each V is S.

Still another embodiment of the invention of Formula Ia and Formula Ia and I is realized when K is H.

Another embodiment of the invention of Formula Ia and I is realized when K is

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A subembodiment of this aspect of the invention is realized when R₅is selected from the group consisting of H, C_1-6alkyl and —(CH₂CH₂O)_pCH₂CH₃. Another subembodiment of this aspect of the invention is realized when R₅is H. Another subembodiment of this aspect of the invention is realized when R₅is C_1-6alkyl. Still another subembodiment of this aspect of the invention is realized when R₅is —(CH₂CH₂O)_pCH₂CH₃.

An embodiment of the invention of Formula Ia and I is realized when any W is independently selected from N and —CH—. Another embodiment of the invention of Formula Ia and I is realized when at least one W is N. Another embodiment of the invention of Formula Ia and I is realized when one to three W is N and the others are —CH—. Another embodiment of the invention of Formula Ia and I is realized when two or more W is —CH—. Another embodiment of the invention of Formula Ia and I is realized when one to three W is —CF—.

An embodiment of the invention of Formula Ia and I is realized when at least one Z is N. Still another embodiment of the invention of Formula Ia and I is realized when each Z is N. Another embodiment of the invention of Formula Ia and I is realized when each Z is —CH—. Another embodiment of the invention of Formula Ia and I is realized when at least one Z is —CH—. Another embodiment of the invention of Formula I is realized when any W and Z within a single nine membered bicycle is independently selected from W═N and Z═—CH—; W═—CH— and Z═N; W═N and Z═N; and W═—CH— and Z is —CH—. Another embodiment of the invention of Formula Ia and I is realized when each W is N and each Z is —CH—. Another embodiment of the invention of the Formula Ia and I is realized when each W is —CH— and each Z is —CH—. Another embodiment of the invention of the Formula Ia and I is realized when each W═—CH— and each Z═N. Another embodiment of the invention of the Formula Ia and I is realized when each W═N and each Z═N.

An embodiment of the invention of Formula Ia and I is realized when any R₁of Formula Ia and I is selected from the group consisting of H, halogen, —OH, —OMe, Me, —OCF₃, —O(CH₂)₂OCH₃, —OCH₂C≡CH, —NH₂,

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Another embodiment of the invention of Formula Ia and I is realized when any R₁is independently selected from the group consisting of H, halogen, —OH, —OMe, -Me, —OCF₃, —O(CH₂)₂OCH₃, —OCH₂C≡CH, and —NH₂. Another embodiment of the invention of Formula Ia and I is realized when any R₁is independently selected from H, halogen, —OH, —OMe, -Me, —OCF₃, —O(CH₂)₂OCH₃, and —NH₂.

An embodiment of the invention of Formula Ia and I is realized when any R₂is independently selected from the group consisting of H, halogen, —OH, —OMe, -Me, and —OCF₃. A subembodiment of this invention of Formula Ia and I is realized when any R₂is independently selected from the group consisting of H and -Me.

An embodiment of the invention of Formula Ia and I is realized when one to three adjacent R₁and R₂may combine to form ═CH₂.

Another embodiment of the invention of Formula Ia and I is realized when any R₃is independently H, halogen, C_1-6alkyl, —C≡CH. A sub-embodiment of the invention Formula 1 is realized when any R₃is independently selected from H, F, Cl, methyl, ethyl, propyl, butyl hexyl and —C≡CH. A subembodiment of the invention of Formula Ia and I is realized when one to three R₁and R₃combine to form —CH₂O— and R₂is hydrogen. A subembodiment of the invention of Formula I is realized when R₃is hydrogen.

An embodiment of the invention of Formula Ia and I is realized when R₄is —CH₂OH. Another embodiment of the invention of Formula Ia and I is realized when R₄is

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A subembodiment of this aspect of the invention is realized when R is H. Another subembodiment of the invention of Formula Ia and I is realized when R is —P(O)(OH)₂. A subembodiment of this aspect of the invention is realized when Q is O. Another subembodiment of this aspect of the invention is realized when Q is —NH—. Another subembodiment of this aspect of the invention is realized when Q is —CH₂—. Another subembodiment of this aspect of the invention is realized when g is 1, R is H and Q is O. Another subembodiment of the invention is realized when g is 2, R is H and Q is O. Still another subembodiment of the invention is realized when g is 3, R is H and Q is O. Yet another subembodiment of this aspect of the invention is realized when g is 1 and Q is —CH₂—, or —NH—. Another subembodiment of the invention is realized when g is 2, and Q is —NH— or —CH₂—. Still another subembodiment of the invention is realized when g is 3, and Q is —NH—, or —CH₂—.

Another embodiment of the invention of Formula Ia and I is realized when R₄is

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Another subembodiment of this aspect of the invention is realized when R is H. Another subembodiment of the invention of Formula Ia and I is realized when R is —P(O)(OH)₂. A subembodiment of this aspect of the invention is realized when Q is O. Another subembodiment of this aspect of the invention is realized when Q is NH. Another subembodiment of this aspect of the invention is realized when Q is CH₂. Another subembodiment of this aspect of the invention is realized when g is 1, R is H and Q is O. Another subembodiment of the invention is realized when g is 2, R is H and Q is O. Still another subembodiment of the invention is realized when g is 3, R is H and Q is O. Yet another subembodiment of this aspect of the invention is realized when g is 1 and Q is —CH₂—, or —NH—. Another subembodiment of the invention is realized when g is 2, and Q is —NH— or —CH₂—. Still another subembodiment of the invention is realized when g is 3, and Q is —NH—, or —CH₂—. Still another embodiment of the invention of Formula Ia and I is realized when R₄is

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An embodiment of the invention of Formula Ia and I is realized when R₄is

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An embodiment of the invention of Formula Ia and I is realized when R₅is H. Another embodiment of the invention of Formula Ia and I is realized when R₅is C_1-6alkyl, wherein alkyl is selected from the group consisting of methyl, ethyl, propyl, butyl, pentyl and hexyl. Another embodiment of the invention of Formula Ia and I is realized when R₅is —(CH₂CH₂O)_pCH₂CH₃. Yet another embodiment of the invention of Formula Ia and I is realized when R₅is

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An embodiment of the invention of Formula Ia and I is realized when R is H. Another embodiment of the invention of Formula Ia and I is realized when R is —P(O)(OH)₂.

An embodiment of the invention of Formula Ia and I is realized when R₄is —CH₂OH and K is H. A subembodiment of this aspect of the invention is realized when Q is O.

Another embodiment of the invention of Formula Ia and I is realized when R₄is —CH₂OH and K is P(O)(OH)₂. A subembodiment of this aspect of the invention is realized when Q is O.

An embodiment of the invention of Formula Ia and I is realized when one or more adjacent R₁and R₂combine to form ═CH₂, and R₄is —CH₂OH. A subembodiment of this aspect of the invention is realized when K is H. Another subembodiment of this aspect of the invention is realized when K is —P(O)(OH)₂.

An embodiment of the invention of Formula Ia and I is realized when one or more adjacent R₁and R₂combine to form ═CH₂; and R₄is selected from the group consisting of

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Still another subembodiment of this aspect of the invention is realized when Q is O. Yet another subembodiment of this aspect of the invention is realized when Q is —NH—, —CH₂—. A subembodiment of this aspect of the invention is realized when K is H. Another subembodiment of this aspect of the invention is realized when K is —P(O)(OH)₂. Another subembodiment of this aspect of the invention is realized when R is H. Another subembodiment of the invention of Formula Ia and I is realized when R is —P(O)(OH)₂.

An embodiment of the invention of Formula Ia and I is realized when R₁and R₃on a sugar moiety are linked to form —CH₂O— and R₄is —CH₂OH. A subembodiment of this aspect of the invention is realized when K is H. Another subembodiment of this aspect of the invention is realized when K is —P(O)(OH)₂.

An embodiment of the invention of Formula Ia and I is realized when R₁and R₃on a sugar moiety are linked to form —CH₂O and R₄is selected from the group consisting of

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A subembodiment of this aspect of the invention is realized when K is H. Another subembodiment of this aspect of the invention is realized when K is —P(O)(OH)₂. Still another subembodiment of this aspect of the invention is realized when Q is O. Yet another subembodiment of this aspect of the invention is realized when Q is independently selected from O, —CH₂— or —NH—, —CH₂—. Another subembodiment of this aspect of the invention is realized when R is H. Another subembodiment of the invention of Formula Ia and I is realized when R is —P(O)(OH)₂.

Another aspect of the invention of Formula Ia is realized when R_xis hydrogen. Still another aspect of the invention of Formula Ia is realized when R_xis —C≡CH.

Another aspect of the invention of Formula Ia is realized when R_sis NH₂. Still another aspect of the invention of Formula Ia is realized when R_sis —C(O)—.

As described herein, unless otherwise indicated, the use of a compound in treatment means that an amount of the compound, generally presented as a component of a formulation that comprises other excipients, is administered in aliquots of an amount, and at time intervals, which provide and maintain at least a therapeutic serum level of at least one pharmaceutically active form of the compound over the time interval between dose administrations.

When any variable (e.g. aryl, heterocycle, R¹, R²etc.) occurs more than one time in any constituent, its definition on each occurrence is independent of every other occurrence. Also, combinations of substituents/or variables are permissible only if such combinations result in stable compounds, is chemically feasible and/or valency permits.

As used herein, unless otherwise specified, the terms in the paragraphs immediately below have the indicated meaning.

“Alkyl” refers to both branched- and straight-chain saturated aliphatic hydrocarbon groups of 1 to 18 carbon atoms, or more specifically, 1 to 12 carbon atoms. Examples of such groups include, but are not limited to, methyl (Me), ethyl (Et), n-propyl (Pr), n-butyl (Bu), n-pentyl, n-hexyl, and the isomers thereof such as isopropyl (i-Pr), isobutyl (i-Bu), sec-butyl (s-Bu), tert-butyl (t-Bu), isopentyl, and isohexyl. Alkyl groups may be optionally substituted with one or more substituents as defined herein. “C_1-6alkyl” refers to an alkyl group as defined herein having 1 to 6 carbon atoms.

“Carboxyl” refers to a monovalent functional group or radical (e.g., COOH) typical of organic acids.

“Halo” or “halogen” refers to fluoro, chloro, bromo or iodo, unless otherwise noted.

The term “haloalkyl” refers to an alkyl in which one or more hydrogen atoms are replaced by halo, and includes alkyl moieties in which all hydrogens have been replaced by halo (e.g., perfluoroalkyl). Alkyl and haloalkyl groups may be optionally inserted with O, N, or S. The terms “aralkyl” refers to an alkyl moiety in which an alkyl hydrogen atom is replaced by an aryl group. Aralkyl includes groups in which more than one hydrogen atom has been replaced by an aryl group. Examples of “aralkyl” include benzyl, 9-fluorenyl, benzhydryl, and trityl groups.

The term “alkenyl” refers to a straight or branched hydrocarbon chain containing 2-8 carbon atoms and characterized in having one or more double bonds. Examples of a typical alkenyl include, but not limited to, allyl, propenyl, 2-butenyl, 3-hexenyl and 3-octenyl groups.

The term “alkynyl” refers to a straight or branched hydrocarbon chain containing 2-8 carbon atoms and characterized in having one or more triple bonds. Some examples of a typical alkynyl are ethynyl, 2-propynyl, and 3-methylbutynyl, and propargyl.

The term “alkoxy” refers to an alky group singularly bonded to oxygen, and the terms “cycloalkoxy” and “arylkoxy” refer to an cyclo alky and aryl alkyl groups singularly bonded to oxygen respectively.

The term “alkylene” refers to a divalent alkyl, e.g., —CH₂—, —CH₂CH₂—, and —CH₂CH₂CH₂—.

The term “cycloalkyl” as employed herein includes saturated cyclic, bicyclic, tricyclic, or polycyclic hydrocarbon groups having 3 to 12 carbons. The cycloalkyl groups herein described may also contain fused rings. Fused rings are rings that share a common carbon-carbon bond or a common carbon atom (e.g., spiro-fused rings). Examples of cycloalkyl moieties include, but are not limited to, cycylopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantyl, norbornyl, and decalin.

The term “heterocyclyl” refers to a nonaromatic 3-10 membered monocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ring system having 1-3 heteroatoms of monocyclic, 1-6 heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms selected from O, N, or S (e.g., carbon atoms and 1-3, 1-6, or 1-9 heteroatoms of N, O, or S if monocyclic, bicyclic, or tricyclic, respectively). The heterocyclyl groups herein described may also contain fused rings. Fused rings are rings that share a common carbon-carbon bond or a common carbon atom (e.g., spiro-fused rings). Examples of heterocyclyl include, but are not limited to tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, morpholino, pyrrolinyl and pyrrolidinyl.

The term “cycloalkenyl” as employed herein includes partially unsaturated, nonaromatic, cyclic, bicyclic, tricyclic, or polycyclic hydrocarbon groups having 5 to 12 carbons, preferably 5 to 8 carbons. The cycloalkenyl groups herein described may also contain fused rings. Fused rings are rings that share a common carbon-carbon bond or a common carbon atom (e.g., spiro-fused rings). Examples of cycloalkenyl moieties include, but are not limited to, cyclohexenyl, and cyclohexadienyl.

The term “heteroaryl” refers to an aromatic 5-8 membered monocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ring system having 1-3 heteroatoms for monocyclic, 1-6heteroatoms for bicyclic, or 1-9 heteroatoms for tricyclic, said heteroatoms selected from O, N, or S (e.g., carbon atoms and 1-3, 1-6, or 1-9 heteroatoms of N, O, or S for monocyclic, bicyclic, or tricyclic, respectively). The heteroaryl groups herein described may also contain fused rings that share a common carbon-carbon bond.

The term “oxo” refers to an oxygen atom, which forms a carbonyl when attached to carbon, an N-oxide when attached to nitrogen, and a sulfoxide or sulfone when attached to sulfur.

The term “acyl” refers to an alkylcarbonyl, cycloalkylcarbonyl, arylcarbonyl, heterocyclylcarbonyl, or heteroarylcarbonyl substituent.

The term “nucleobase” refers to a nitrogen-containing heterocyclic moiety, which are the parts of the nucleic acids that are involved in the hydrogen-bonding and bind one nucleic acid strand to another complementary strand in a sequence specific manner. The most common naturally-occurring nucleobases are: adenine (A), cyctosine (C), guanine (G), thymine (T), and uracil (U).

The term “modified nucleobase” refers to a moiety that can replace a nucleobase. The modified nucleobase mimics the spatial arrangement, electronic properties, or some other physiochemical property of the nucleobase and retains the property of the hydrogen-bonding that binds one nucleic acid strand to another in a sequence specific manner. A modified nucleobase can pair with at least one of the five naturally occurring bases (uracil, thymine, adenine, cytosine, and guanine) without substantially affecting the melting behavior, recognition by intracellular enzymes, or activity of the oligonucleotide duplex. The term “modified nucleoside” or “modified nucleotide” refers to a nucleoside or nucleotide that contains a modified nucleobase and/or other chemical modification disclosed herein, such as modified sugar, modified phosphorus atom bridges or modified internucleoside linkages. Non-limiting examples of nucleobases include uracil, thymine, adenine, cytosine, and guanine optionally having their respective amino groups protected by, e.g., acyl protecting groups, 5-propynyl-uracil, 2-thio-5-propynyl-uracil, 5-methylcytosine, 2-fluorouracil, 2-fluorocytosine, 5-bromouracil, 5-iodouracil, 2,6-diaminopurine, azacytosine, 2-thiouracil, 2-thiothymine, 2-aminopurine, N9-(2-amino-6-chloropurine, N9-(2,6-diaminopurine), hypoxanthine, N9-(7-deaza-guanine), N9-(7-deaza-8-aza-guanine), N8-(8-aza-7-deazaadenine), pyrimidine analogs such as pseudoisocytosine and pseudouracil and other modified nucleobases such as 8-substituted purines, xanthine, or hypoxanthine.

The terms “adeninyl, cytosinyl, guaninyl, thyminyl, and uracilyl” and the like refer to radicals of adenine, cytosine, guanine, thymine, and uracil.

The term cholesterol is presented by

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and when used as a radical, it is linked to the core structure through bond having the “OH” group.

A “protected” moiety refers to a reactive functional group, e.g., a hydroxyl group or an amino group, or a class of molecules, e.g., sugars, having one or more functional groups, in which the reactivity of the functional group is temporarily blocked by the presence of an attached protecting group. Protecting groups useful for the monomers and methods described herein can be found, e.g., in Greene, T. W., Protective Groups in Organic Synthesis (John Wiley and Sons: New York), 1981, which is hereby incorporated by reference.

A further aspect of the present invention relates to a pharmaceutical composition comprising a modified oligonucleotide as defined herein.

The pharmaceutical composition according to the invention may further comprise pharmaceutically acceptable carriers, diluents, and/or adjuvants. The term “carrier” when used herein includes carriers, excipients and/or stabilisers that are non-toxic to the cell or mammal being exposed thereto at the dosages and concentrations employed. Often the physiologically acceptable carriers are aqueous pH buffered solutions or liposomes. Examples of physiologically acceptable carriers include buffers such as phosphate, citrate and other organic acids (however, with regard to the formulation of the present invention, a phosphate buffer is preferred); anti-oxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinyl pyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose or dextrins, gelating agents such as EDTA, sugar, alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and/or non-ionic surfactants such as TWEEN, polyethylene or polyethylene glycol. According to an embodiment the compound of the invention is dissolved in sterile deionized water.

Such a composition and/or formulation according to the invention can be administered to a subject in need thereof, particularly a human patient, in a sufficient dose for the treatment of the specific conditions by suitable means. For example, the composition and/or formulation according to the invention may be formulated as a pharmaceutical composition together with pharmaceutically acceptable carriers, diluents and/or adjuvants. Therapeutic efficiency and toxicity may be determined according to standard protocols. The pharmaceutical composition may be administered intratumorally, and systemically, e.g., intraperitoneally, intramuscularly, intravenously, locally, such as intranasally, subcutaneously, intradermally or intrathecally. The dose of the composition and/or formulation administered will, of course, be dependent on the subject to be treated and on the condition of the subject, such as the subject's weight, the subject's age and the type and severity of the disease or injury to be treated, the manner of administration and the judgement of the prescribing physician.

In an embodiment, the pharmaceutical composition is administered intradermally. In another embodiment, the composition is administered intradermally via tattooing, microneedling and/or microneedle patches.

Methods of Use

Compounds described herein having therapeutic applications, such as the compounds of general Formula Ia and Formula I, and the compounds of Examples 1 through 838, can be administered to a patient for the purpose of inducing an immune response, inducing a RIG-I-dependent IFN production and/or inducing anti-tumor activity. In one embodiment, the present invention provides for both prophylactic and therapeutic methods of inducing an IFN response in a patient. The term “administration” and variants thereof (e.g., “administering” a compound) means providing the compound to the individual in need of treatment. When a compound is provided in combination with one or more other active agents (e.g., antiviral agents useful for treating HCV infection or anti-tumor agents for treating cancers), “administration” and its variants are each understood to include concurrent and sequential provision of the compound or salt and other agents.

The present invention encompasses the use of the short oligonucleotide hairpin molecule of the invention to prevent and/or treat any disease, disorder, or condition in which inducing IFN production would be beneficial. For example, increased IFN production, by way of the nucleic acid molecule of the invention, may be beneficial to prevent or treat a wide variety of disorders, including, but not limited to, bacterial infection, viral infection, parasitic infection, immune disorders, respiratory disorders, cancer and the like.

Infections include, but are not limited to, viral infections, bacterial infections, anthrax, parasitic infections, fungal infections and prion infection.

Viral infections include, but are not limited to, infections by hepatitis C, hepatitis B, influenza virus, herpes simplex virus (HSV), human immunodeficiency virus (HIV), respiratory syncytial virus (RSV), vesicular stomatitis virus (VSV), cytomegalovirus (CMV), poliovirus, encephalomyocarditis virus (EMCV), human papillomavirus (HPV) and smallpox virus. In one embodiment, the infection is an upper respiratory tract infection caused by viruses and/or bacteria, in particular, flu, more specifically, bird flu.

Bacterial infections include, but are not limited to, infections by streptococci, staphylococci, E. coli, and Pseudomonas. In one embodiment, the bacterial infection is an intracellular bacterial infection which is an infection by an intracellular bacterium such as mycobacteria (tuberculosis), chlamydia, mycoplasma, listeria, and an facultative intracellular bacterium such as Staphylococcus aureus.

Parasitic infections include, but are not limited to, worm infections, in particular, intestinal worm infection, microeukaryotes, and vector-borne diseases, including for example Leishmaniasis.

In a preferred embodiment, the infection is a viral infection or an intracellular bacterial infection. In a more preferred embodiment, the infection is a viral infection by hepatitis C, hepatitis B, influenza virus, RSV, HPV, HSV1, HSV2, and CMV.

Immune disorders include, but are not limited to, allergies, autoimmune disorders, and immunodeficiencies.

Allergies include, but are not limited to, respiratory allergies, contact allergies and food allergies.

Autoimmune diseases or disorders include, but are not limited to, multiple sclerosis, diabetes mellitus, arthritis (including rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, psoriatic arthritis), encephalomyelitis, myasthenia gravis, systemic lupus erythematosis, autoimmune thyroiditis, dermatisis (including atopic dermatitis and eczematous dermatitis), psoriasis, Siogren's Syndrome, Crohn's Disease, aphthous ulcer, iritis, conjunctivitis, keratoconjunctivitis, ulcerative colitis, asthma, allergic asthma, cutaneous lupus erythematosus, scleroderma, vaginitis, proctitis, drug eruptions, leprosy reversal reactions, erythema nodosum leprosum, autoimmune uveitis, allergic encephalomyelitis, acute necrotizing hemorrhagic encephalopathy, idiopathic bilateral progressive sensorineural hearing loss, aplastic anemia, pure red cell anemia, idiopathic thrombocytopenia, polychondritis, Wegener's granulomatosis, chronic active hepatitis, Stevens-Johnson syndrome, idiopathic sprue, lichen planus, Graves' disease, sarcoidosis, primary biliary cirrhosis, uveitis posterior, and interstitial lung fibrosis.

Immunodeficiencies include, but are not limited to, spontaneous immunodeficiency, acquired immunodeficiency (including AIDS), drug-induced immunodeficiency or immunosuppression (such as that induced by immunosuppressants used in transplantation and chemotherapeutic agents used for treating cancer), and immunosuppression caused by chronic hemodialysis, trauma or surgical procedures.

Respiratory disorders include, but are not limited to, acute lung injury (ALI), acute respiratory distress syndrome (ARDS), asthma, chronic obstructive pulmonary disease (COPD), obstructive sleep apnea (OSA), idiopathic pulmonary fibrosis (IPF), tuberculosis, pulmonary hypertension, pleural effusion, and lung cancer.

Examples of cancers include, but are not limited to, Acute Lymphoblastic Leukemia; Acute Myeloid Leukemia; Adrenocortical Carcinoma; AIDS-Related Lymphoma; AIDS-Related Malignancies; Anal Cancer; Astrocytoma; Bile Duct Cancer; Bladder Cancer; Bone Cancer, Osteosarcoma/Malignant Fibrous Histiocytoma; Brain Stem Glioma; Brain Tumor, Cerebellar Astrocytoma; Brain Tumor, Cerebral Astrocytoma/Malignant Glioma; Brain Tumor, Ependymoma; Brain Tumor, Medulloblastoma; Brain Tumor, Supratentorial Primitive Neuroectodermal Tumors; Brain Tumor, Visual Pathway and Hypothalamic Glioma; Breast Cancer; Bronchial Adenomas/Carcinoids; Carcinoid Tumor; Carcinoid Tumor, Gastrointestinal; Carcinoma, Adrenocortical; Carcinoma, Islet Cell; Central Nervous System Lymphoma, Primary; Cerebral Astrocytoma/Malignant Glioma; Cervical Cancer; Chronic Lymphocytic Leukemia; Chronic Myelogenous Leukemia; Chronic Myeloproliferative Disorders; Clear Cell Sarcoma of Tendon Sheaths; Colon Cancer; Colorectal Cancer; Cutaneous T-Cell Lymphoma; Endometrial Cancer; Ependymoma; Epithelial Cancer, Ovarian; Esophageal Cancer; Esophageal Cancer; Ewing's Family of Tumors; Extracranial Germ Cell Tumor; Extrahepatic Bile Duct Cancer; Eye Cancer, Intraocular Melanoma; Eye Cancer, Retinoblastoma; Gallbladder Cancer; Gastric (Stomach) Cancer; Gastrointestinal Carcinoid Tumor; Germ Cell Tumor, Extracranial, Childhood; Germ Cell Tumor, Extragonadal; Germ Cell Tumor, Ovarian; Gestational Trophoblastic Tumor; Glioma, Childhood Brain Stem; Glioma, Childhood Visual Pathway and Hypothalamic; Hairy Cell Leukemia; Head and Neck Cancer; Hepatocellular (Liver) Cancer; Hodgkin's Lymphoma; Hypopharyngeal Cancer; Hypothalamic and Visual Pathway Glioma; Intraocular Melanoma; Islet Cell Carcinoma (Endocrine Pancreas); Kaposi's Sarcoma; Kidney Cancer; Laryngeal Cancer; Leukemia, Acute Lymphoblastic; Leukemia, Acute Myeloid; Leukemia, Chronic Lymphocytic; Leukemia, Chronic Myelogenous; Leukemia, Hairy Cell; Lip and Oral Cavity Cancer; Liver Cancer; Lung Cancer, Non-Small Cell; Lung Cancer, Small Cell; Lymphoblastic Leukemia; Lymphoma, AIDS-Related; Lymphoma, Central Nervous System (Primary); Lymphoma, Cutaneous T-Cell; Lymphoma, Hodgkin's; Lymphoma, Hodgkin's During Pregnancy; Lymphoma, Non-Hodgkin's; Lymphoma, Primary Central Nervous System; Macroglobulinemia, Waldenstrom's; Male Breast Cancer; Malignant Mesothelioma; Malignant Thymoma; Medulloblastoma, Childhood; Melanoma; Melanoma, Intraocular; Merkel Cell Carcinoma; Mesothelioma, Malignant; Metastatic Squamous Neck Cancer with Occult Primary; Multiple Endocrine Neoplasia Syndrome, Childhood; Multiple Myeloma/Plasma Cell Neoplasm; Mycosis Fungoides; Myelodysplastic Syndromes; Myelogenous Leukemia, Chronic; Myeloid Leukemia; Myeloma, Multiple; Myeloproliferative Disorders, Chronic; Nasal Cavity and Paranasal Sinus Cancer; Nasopharyngeal Cancer; Neuroblastoma; Non-Hodgkin's Lymphoma; Non-Small Cell Lung Cancer; Oral Cancer; Oral Cavity and Lip Cancer; Oropharyngeal Cancer; steosarcoma/Malignant Fibrous Histiocytoma of Bone; Ovarian Epithelial Cancer; Ovarian Germ Cell Tumor; Ovarian Low Malignant Potential Tumor; Pancreatic Cancer; Paranasal Sinus and Nasal Cavity Cancer; Parathyroid Cancer; Penile Cancer; Pheochromocytoma; Pineal and Supratentorial Primitive Neuroectodermal Tumors; Pituitary Tumor; Plasma Cell Neoplasm/Multiple Myeloma; Pleuropulmonary Blastoma; Pregnancy and Breast Cancer; Pregnancy and Hodgkin's Lymphoma; Pregnancy and Non-Hodgkin's Lymphoma; Primary Central Nervous System Lymphoma; Primary Liver Cancer; Prostate Cancer; Rectal Cancer; Renal Cell (Kidney) Cancer; Renal Pelvis and Ureter, Transitional Cell Cancer; Retinoblastoma; Rhabdomyosarcoma; Salivary Gland Cancer; Sarcoma, Ewing's Family of Tumors; Sarcoma, Kaposi's; Sarcoma (Osteosarcoma)/Malignant Fibrous Histiocytoma of Bone; Sarcoma, Soft Tissue; Sezary Syndrome; Skin Cancer; Skin Cancer (Melanoma); Skin Carcinoma, Merkel Cell; Small Cell Lung Cancer; Small Intestine Cancer; Soft Tissue Sarcoma; Squamous Neck Cancer with Occult Primary, Metastatic; Stomach (Gastric) Cancer; Supratentorial Primitive Neuroectodermal Tumors; T-Cell Lymphoma, Cutaneous; Testicular Cancer; Thymoma, Malignant; Thyroid Cancer; Transitional Cell Cancer of the Renal Pelvis and Ureter; Trophoblastic Tumor, Gestational; Ureter and Renal Pelvis, Transitional Cell Cancer; Urethral Cancer; Uterine Sarcoma; Vaginal Cancer; Visual Pathway and Hypothalamic Glioma; Vulvar Cancer; Waldenstrom's Macro globulinemia; and Wilms' Tumor.

In one embodiment, the cancer is brain cancer, such as an astrocytic tumor (e.g., pilocytic astrocytoma, subependymal giant-cell astrocytoma, diffuse astrocytoma, pleomorphic xanthoastrocytoma, anaplastic astrocytoma, astrocytoma, giant cell glioblastoma, glioblastoma, secondary glioblastoma, primary adult glioblastoma, and primary pediatric glioblastoma); oligodendroglial tumor (e.g., oligodendroglioma, and anaplastic oligodendroglioma); oligoastrocytic tumor (e.g., oligoastrocytoma, and anaplastic oligoastrocytoma); ependymoma (e.g., myxopapillary ependymoma, and anaplastic ependymoma); medulloblastoma; primitive neuroectodermal tumor, schwannoma, meningioma, meatypical meningioma, anaplastic meningioma; and pituitary adenoma. In another embodiment, the brain cancer is glioma, glioblastoma multiforme, paraganglioma, or suprantentorial primordial neuroectodermal tumors (sPNET).

In another embodiment, the cancer is leukemia, such as acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), chronic myelogenous leukemia (CML), myeloproliferative neoplasm (MPN), post-MPN AML, post-MDS AML, del(5q)-associated high risk MDS or AML, blast-phase chronic myelogenous leukemia, angioimmunoblastic lymphoma, and acute lymphoblastic leukemia.

In one embodiment, the cancer is skin cancer, including melanoma. In another embodiment, the cancer is prostate cancer, breast cancer, thyroid cancer, colon cancer, or lung cancer. In another embodiment, the cancer is sarcoma, including central chondrosarcoma, central and periosteal chondroma, and fibrosarcoma. In another embodiment, the cancer is cholangiocarcinoma.

Administration and Dosing

As used herein, the terms “treatment” and “treating” refer to all processes wherein there may be a slowing, interrupting, arresting, controlling, or stopping of the progression of a disease or disorder described herein. The terms do not necessarily indicate a total elimination of all disease or disorder symptoms. The terms also include the potential prophylactic therapy of the mentioned conditions, particularly in a subject that is predisposed to such disease or disorder.

The terms “administration of” and or “administering” a compound should be understood to include providing a compound described herein, or a pharmaceutically acceptable salt thereof, and compositions of the foregoing to a subject.

The amount of a compound administered to a subject is an amount sufficient to induce an immune response and/or to induce RIG-I-dependent type I interferon production in the subject. In an embodiment, the amount of a compound can be an “effective amount” or “therapeutically effective amount,” wherein the subject compound is administered in an amount that will elicit a biological or medical response of a tissue, system, animal or human that is being sought by a researcher, veterinarian, medical doctor or other clinician. An effective amount does not necessarily include considerations of toxicity and safety related to the administration of a compound.

An effective amount of a compound will vary with the particular compound chosen (e.g., considering the potency, efficacy, and/or half-life of the compound); the route of administration chosen; the condition being treated; the severity of the condition being treated; the age, size, weight, and physical condition of the subject being treated; the medical history of the subject being treated; the duration of the treatment; the nature of a concurrent therapy; the desired therapeutic effect; and like factors and can be routinely determined by the skilled artisan.

Biological methods for introducing a nucleic acid into a host cell include the use of DNA and RNA vectors. Viral vectors, and especially retroviral vectors, have become the most widely used method for inserting genes into mamma-lian, e.g., human cells.

The compounds disclosed herein may be administered by any suitable route including oral and parenteral administration. Parenteral administration is typically by injection or infusion and includes intravenous, intramuscular, and subcutaneous injection or infusion.

The compounds disclosed herein may be administered once or according to a dosing regimen wherein a number of doses are administered at varying intervals of time for a given period of time. For example, doses may be administered one, two, three, or four times per day. Doses may be administered until the desired therapeutic effect is achieved or indefinitely to maintain the desired therapeutic effect. Suitable dosing regimens for a compound disclosed herein depend on the pharmacokinetic properties of that compound, such as absorption, distribution and half-life which can be determined by a skilled artisan. In addition, suitable dosing regimens, including the duration such regimens are administered, for a compound disclosed herein depend on the disease or condition being treated, the severity of the disease or condition, the age and physical condition of the subject being treated, the medical history of the subject being treated, the nature of concurrent therapy, the desired therapeutic effect, and like factors within the knowledge and expertise of the skilled artisan. It will be further understood by such skilled artisans that suitable dosing regimens may require adjustment given an individual subject's response to the dosing regimen or over time as the individual subject needs change. Typical daily dosages may vary depending upon the particular route of administration chosen. Typical daily dosages for oral administration, to a human weighing approximately 70 kg would range from about 0.1 mg to about 2 g, or more specifically, 0.1 mg to 500 mg, or even more specifically, 0.2 mg to 100 mg, of a compound of general Formula Ia and Formula I.

One embodiment of the present disclosure provides for a method of treating a cell proliferation disorder comprising administration of an effective amount of a compound of general Formula Ia and Formula I to a subject in need of treatment thereof. In one embodiment, the cell proliferation disorder is cancer.

In one embodiment, the cancer is brain cancer, leukemia, skin cancer, prostate cancer, thyroid cancer, colon cancer, lung cancer or sarcoma. In another embodiment the cancer is glioma, glioblastoma multiforme, paraganglioma, suprantentorial primordial neuroectodermal tumors, acute myeloid leukemia, myelodysplastic syndrome, chronic myelogenous leukemia, melanoma, breast, prostate, thyroid, colon, lung, central chondrosarcoma, central and periosteal chondroma tumors, fibrosarcoma, and cholangiocarcinoma.

In one embodiment, disclosed herein is the use of a compound of general Formula Ia and Formula I in a therapy. The compound may be useful in a method of inducing an immune response and/or inducing RIG-I-dependent type I interferon production in a subject, such as a mammal in need of such inhibition, comprising administering an effective amount of the compound to the subject.

In one embodiment, disclosed herein is a pharmaceutical composition comprising at least one compound of general Formula Ia and Formula I, or a pharmaceutically acceptable salt thereof, for use in potential treatment to induce an immune response and/or to induce RIG-I-dependent type I interferon production.

In one embodiment, disclosed herein is the use of a compound of general Formula Ia and Formula I in the manufacture of a medicament for the treatment to induce an immune response and/or to induce RIG-I-dependent type I interferon production. In one embodiment, the disease or disorder to be treated is a cell proliferation disorder. In another embodiment, the cell proliferation disorder is cancer. In another embodiment, the cancer is brain cancer, leukemia, skin cancer, breast, prostate cancer, thyroid cancer, colon cancer, lung cancer, or sarcoma. In another embodiment, the cancer is glioma, glioblastoma multiforme, paraganglioma, suprantentorial primordial neuroectodermal tumors, acute myeloid leukemia, myelodysplastic syndrome, chronic myelogenous leukemia, melanoma, breast, prostate, thyroid, colon, lung, central chondrosarcoma, central and periosteal chondroma tumors, fibrosarcoma, and/or cholangiocarcinoma.

Compositions

The present invention provides a pharmaceutical composition comprising at least one nucleic acid molecule of the present invention and a pharmaceutically acceptable carrier. The formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of bringing the active ingredient into association with a carrier or one or more other accessory ingredients, and then, if necessary or desirable, shaping or packaging the product into a desired single- or multi-dose unit.

The term “composition” as used herein is intended to encompass a dosage form comprising a specified compound in a specified amount, as well as any dosage form which results, directly or indirectly, from combination of a specified compound in a specified amount. Such term is intended to encompass a dosage form comprising a compound of general Formula Ia and Formula I, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers or excipients. Accordingly, the compositions of the present disclosure encompass any composition made by admixing a compound of the present disclosure and one or more pharmaceutically acceptable carrier or excipients. By “pharmaceutically acceptable”, it is meant the carriers or excipients are compatible with the compound disclosed herein and with other ingredients of the composition.

For the purpose of inducing an immune response and/or inducing a RIG-I-dependent type I interferon production, the compounds, optionally in the form of a salt, can be administered by means that produce contact of the active agent with the agent's site of action. They can be administered by conventional means available for use in conjunction with pharmaceuticals, either as individual therapeutic agents or in a combination of therapeutic agents. They can be administered alone, but typically are administered with a pharmaceutical carrier selected on the basis of the chosen route of administration and standard pharmaceutical practice.

An embodiment of the invention is realized by a composition comprising a nucleic acid capable of inducing interferon production, wherein the molecule comprises a single strand of 24 bases by 5′ GCUCAUUUACCAGCGUAAAUGAGC 3′ (SEQ ID NO: 841), said single strand having a double-stranded section of less than 12 base pairs. A subembodiment of this aspect of the invention is realized when the single stranded nucleic acid forms a hairpin structure comprising the double-stranded section and a loop. Another subembodiment of this aspect of the invention is realized when the double-stranded nucleic acid comprises one or two blunt ends. Another subembodiment of this aspect of the invention is realized when

the double-stranded nucleic acid comprises a 5′ blunt end. Another subembodiment of this aspect of the invention is realized when the nucleic acid comprises at least one of the group consisting of a 5′-OH, 5′-triphosphate and a 5′-diphosphate. Another subembodiment of this aspect of the invention is realized when the molecule comprises a modified phosphodiester backbone. Still another subembodiment of this aspect of the invention is realized when the molecule comprises at least one 2′-modified nucleotide. A further aspect of this embodiment of the invention is realized when the 2′-modified nucleotide comprises a modification selected from the group consisting of: 2′-OH, 2′-methyl, 2′-OCF₃, 2′-OCH₂C≡CH, —NH₂, 2′-deoxy, 2′-deoxy-2′-fluoro, 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), 2′-O-dimethylaminoethyl-oxyethyl (2′-O-DMAEOE), and 2′-O—N-methylacetamido (2′-O-NMA). Another subembodiment of this aspect of the invention is realized when the molecule comprises at least one modified phosphate group. Another subembodiment of this aspect of the invention is realized when the molecule comprises at least one modified base. Another subembodiment of this aspect of the invention is realized when the double stranded section comprises one or more mispaired bases.

The compositions of the disclosure are prepared using techniques and methods known to those skilled in the art. Some methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).

In one embodiment, disclosed herein is a composition comprising a compound of general Formula Ia and Formula I or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers or excipients. The composition may be prepared and packaged in bulk form wherein an effective amount of a compound of the disclosure can be extracted and then given to a subject, such as with powders or syrups. Alternatively, the composition may be prepared and packaged in unit dosage form wherein each physically discrete unit contains an effective amount of a compound of general Formula Ia and Formula I. When prepared in unit dosage form, the composition of the disclosure typically contains from about 0.1 mg to 2 g, or more specifically, 0.1 mg to 500 mg, or even more specifically, 0.2 mg to 100 mg, of a compound of general Formula Ia and Formula I, or a pharmaceutically acceptable salt thereof.

The pharmaceutical composition according to the invention may further comprise pharmaceutically acceptable carriers, diluents, and/or adjuvants. The term “carrier” when used herein includes excipients and/or stabilisers that are non-toxic to the cell or mammal being exposed thereto at the dosages and concentrations employed. Often the physiologically acceptable carriers are aqueous pH buffered solutions or liposomes. Suitable pharmaceutically acceptable carriers or excipients include the following non-limiting types: diluents, lubricants, binders, disintegrants, fillers, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavoring agents, flavor masking agents, coloring agents, anti-caking agents, hemectants, chelating agents, plasticizers, viscosity increasing agents, antioxidants, preservatives, stabilizers, surfactants, and buffering agents. Particular examples of physiologically acceptable carriers include buffers such as phosphate, citrate and other organic acids; anti-oxidants including ascorbic acid, low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatine or immunoglobulins; hydrophilic polymers such as polyvinyl pyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose or dextrins; gelating agents such as EDTA, sugar, alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and/or non-ionic surfactants such as TWEEN, polyethylene or polyethylene glycol. In another embodiment the compound of the invention is dissolved in sterile deionized water.

A skilled artisan possesses the knowledge and skill in the art to select suitable pharmaceutically acceptable carriers and excipients in appropriate amounts for the use in the disclosure. In addition, there are a number of resources available to the skilled artisan, which describe pharmaceutically acceptable carriers and excipients and may be useful in selecting suitable pharmaceutically acceptable carriers and excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).

The compounds disclosed herein and a pharmaceutically acceptable carrier or excipient(s) will typically be formulated into a dosage form adapted for administration to a subject by a desired route of administration. Many systems for administering active substances into cells are already known and include lipid nanoparticles (LNPs), emulsions, liposomes, exosomes, dendrimers, natural polymers, e.g., chitosan, polyethylenimines (PEIs), immuno- and ligand complexes, micelles, microparticles and/or polymeric nanoparticles and cyclodextrins (see, Drug Transport in Antimicrobial and Anticancer Chemotherapy. G. Papadakou Ed., CRC Press, 1995 and US2013/0037977). These systems may be adapted for (1) oral administration, such as tablets, capsules, caplets, pills, troches, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets, and cachets; and (2) parenteral administration, such as sterile solutions, suspensions, and powders for reconstitution. Suitable pharmaceutically acceptable carriers or excipients will vary depending upon the particular dosage form chosen. In addition, suitable pharmaceutically acceptable carriers or excipients may be chosen for a particular function that they may serve in the composition. For example, certain pharmaceutically acceptable carriers or excipients may be chosen for their ability to facilitate the production of uniform dosage forms. Certain pharmaceutically acceptable carriers or excipients may be chosen for their ability to facilitate the production of stable dosage forms. Certain pharmaceutically acceptable carriers or excipients may be chosen for their ability to facilitate the carrying or transporting of a compound disclosed herein, once administered to the subject, from one organ or portion of the body to another organ or another portion of the body. Certain pharmaceutically acceptable carriers or excipients may be chosen for their ability to enhance patient compliance.

In an embodiment, the disclosure is directed to a solid oral dosage form such as a tablet or capsule comprising an effective amount of a compound of the disclosure and a diluent or filler. Suitable diluents and fillers include lactose, sucrose, dextrose, mannitol, sorbitol, starch (e.g., corn starch, potato starch, and pre-gelatinized starch), cellulose and its derivatives, (e.g., microcrystalline cellulose), calcium sulfate, and dibasic calcium phosphate. The oral solid dosage form may further comprise a binder. Suitable binders include starch (e.g., corn starch, potato starch, and pre-gelatinized starch) gelatin, acacia, sodium alginate, alginic acid, tragacanth, guar gum, povidone, and cellulose and its derivatives (e.g., microcrystalline cellulose). The oral solid dosage form may further comprise a disintegrant. Suitable disintegrants include crospovidone, sodium starch glycolate, croscarmelose, alginic acid, and sodium carboxymethyl cellulose. The oral solid dosage form may further comprise a lubricant. Suitable lubricants include stearic acid, magnesium stearate, calcium stearate, and talc.

Where appropriate, dosage unit formulations for oral administration can be microencapsulated. The composition can also be prepared to prolong or sustain the release as, for example, by coating or embedding particulate material in polymers, wax, or the like.

The compounds disclosed herein may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyrancopolymer, polyhydroxypropylmethacrylamidephenol, polyhydroxyethylaspartamidephenol, or polyethyleneoxidepolylysine substituted with palmitoyl residues. Furthermore, the compounds of the disclosure may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example polylactic acid, polepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanacrylates and cross-linked or amphipathic block copolymers of hydrogels.

In one embodiment, the disclosure is directed to a liquid oral dosage form. Oral liquids such as solution, syrups and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of a compound disclosed herein. Syrups can be prepared by dissolving the compound of the disclosure in a suitably flavored aqueous solution; while elixirs are prepared through the use of a non-toxic alcoholic vehicle. Suspensions can be formulated by dispersing a compound disclosed herein in a non-toxic vehicle. Solubilizers and emulsifiers such as ethoxylated isostearyl alcohols and polyoxy ethylene sorbitol ethers, preservatives, flavor additives such as peppermint oil or other natural sweeteners or saccharin or other artificial sweeteners and the like can also be added.

In another embodiment, the disclosure is directed to compositions for parenteral administration. Compositions adapted for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions that may include suspending agents and thickening agents. The compositions may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets.

Combinations

The compounds of general Formula Ia and Formula I may be administered in combination with one or more additional therapeutic agents. In embodiments, one or more compound of general Formula Ia and Formula I may be co-adminstered. The additional therapeutic agent(s) may be administered in a single dosage form with the compound of general Formula Ia and Formula I, or the additional therapeutic agent(s) may be administered in separate dosage form(s) from the dosage form containing the compound of general Formula Ia and Formula I. The additional therapeutic agent(s) may be one or more agents selected from the group consisting of anti-viral compounds, antigens, adjuvants, anti-cancer agents, CTLA-4, LAG-3 and PD-1 pathway antagonists, lipids, peptides, chemotherapeutic agents, immunomodulatory cell lines, checkpoint inhibitors, vascular endothelial growth factor (VEGF) receptor inhibitors, topoisomerase II inhibitors, smoothen inhibitors, alkylating agents, anti-tumor antibiotics, anti-metabolites, retinoids, and immunomodulatory agents including but not limited to anti-cancer vaccines. It will be understood the descriptions of the above additional therapeutic agents may be overlapping. It will also be understood that the treatment combinations are subject to optimization, and it is understood that the best combination to use of the compounds of general Formula Ia and Formula I and one or more additional therapeutic agents will be determined based on the individual patient needs.

A compound disclosed herein may be used in combination with one or more other active agents, including but not limited to, other anti-cancer agents that are used in the prevention, treatment, control, amelioration, or reduction of risk of a particular disease or condition (e.g., cell proliferation disorders). In one embodiment, a compound disclosed herein is combined with one or more other anti-cancer agents for use in the prevention, treatment, control amelioration, or reduction of risk of a particular disease or condition for which the compounds disclosed herein are useful. Such other active agents may be administered, by a route and in an amount commonly used therefor, contemporaneously or sequentially with a compound of the present disclosure.

When a compound disclosed herein is used contemporaneously with one or more other active agents, a composition containing such other active agents in addition to the compound disclosed herein is contemplated. Accordingly, the compositions of the present disclosure include those that also contain one or more other active ingredients, in addition to a compound disclosed herein. A compound disclosed herein may be administered either simultaneously with, or before or after, one or more other therapeutic agent(s). A compound disclosed herein may be administered separately, by the same or different route of administration, or together in the same pharmaceutical composition as the other agent(s).

Products provided as a combined preparation include a composition comprising a compound of general Formula Ia and Formula I and one or more other active agent(s) together in the same pharmaceutical composition, or a compound of general Formula Ia and Formula I and one or more other therapeutic agent(s) in separate form, e.g. in the form of a kit.

The weight ratio of a compound disclosed herein to a second active agent may be varied and will depend upon the effective dose of each agent. Generally, an effective dose of each will be used. Thus, for example, when a compound disclosed herein is combined with another agent, the weight ratio of the compound disclosed herein to the other agent will generally range from about 1000:1 to about 1:1000, such as about 200:1 to about 1:200. Combinations of a compound disclosed herein and other active agents will generally also be within the aforementioned range, but in each case, an effective dose of each active agent should be used. In such combinations, the compound disclosed herein and other active agents may be administered separately or in conjunction. In addition, the administration of one element may be prior to, concurrent to, or subsequent to the administration of other agent(s).

In one embodiment, this disclosure provides a composition comprising a compound of general Formula Ia and Formula I and at least one other therapeutic agent as a combined preparation for simultaneous, separate or sequential use in therapy. In one embodiment, the therapy is the treatment of a cell proliferation disorder, such as cancer.

In one embodiment, the disclosure provides a kit comprising two or more separate pharmaceutical compositions, at least one of which contains a compound of general Formula Ia and Formula I. In one embodiment, the kit comprises means for separately retaining said compositions, such as a container, divided bottle, or divided foil packet. An example of such a kit is a blister pack, as typically used for the packaging of tablets, capsules, and the like.

A kit of this disclosure may be used for administration of different dosage forms, for example, oral and parenteral, for administration of the separate compositions at different dosage intervals, or for titration of the separate compositions against one another. To assist with compliance, a kit of the disclosure typically comprises directions for administration.

Disclosed herein is a use of a compound of general Formula Ia and Formula I for treating a cell proliferation disorder, wherein the medicament is prepared for administration with another active agent. The disclosure also provides the use of another active agent for treating a cell proliferation disorder, wherein the medicament is administered with a compound of general Formula Ia and Formula I.

The disclosure also provides the use of a compound of general Formula Ia and Formula I for treating a cell proliferation disorder, wherein the patient has previously (e.g., within 24 hours) been treated with another active agent. The disclosure also provides the use of another therapeutic agent for treating a cell proliferation disorder, wherein the patient has previously (e.g., within 24 hours) been treated with a compound of general Formula Ia and Formula I. The second agent may be applied a week, several weeks, a month, or several months after the administration of a compound disclosed herein.

Anti-viral compounds that may be used in combination with the compounds of general Formula Ia and Formula I disclosed herein include hepatitis B virus (HBV) inhibitors, hepatitis C virus (HCV) protease inhibitors, HCV polymerase inhibitors, HCV NS4A inhibitors, HCV NS5A inhibitors, HCV NS5b inhibitors, and human immunodeficiency virus (HIV) inhibitors.

Antigens and adjuvants that may be used in combination with the compounds of general Formula Ia and Formula I disclosed herein include B7 costimulatory molecule, interleukin-2, interferon-γ, GM-CSF, CTLA-4 antagonists, OX-40/OX-40 ligand, CD40/CD40 ligand, sargramostim, levamisol, vaccinia virus, Bacille Calmette-Guerin (BCG), liposomes, alum, Freund's complete or incomplete adjuvant, detoxified endotoxins, mineral oils, surface active substances such as lipolecithin, pluronic polyols, polyanions, peptides, and oil or hydrocarbon emulsions. Adjuvants, such as aluminum hydroxide or aluminum phosphate, can be added to increase the ability of the compound to trigger, enhance, or prolong an immune response. Additional materials, such as cytokines, chemokines, and bacterial nucleic acid sequences, like CpG, a toll-like receptor (TLR) 9 agonist as well as additional agonists for TLR 2, TLR 4, TLR 5, TLR 7, TLR 8, TLR9, including lipoprotein, LPS, monophosphoryllipid A, lipoteichoic acid, imiquimod, resiquimod, stimulator of interferon genes (STING) agonists and in addition retinoic acid-inducible gene I (RIG-I) agonists such as poly I:C, used separately or in combination with the described compositions are also potential adjuvants.

CLTA-4 and PD-1 pathways are important negative regulators of immune response. Activated T-cells upregulate CTLA-4, which binds on antigen-presenting cells and inhibits T-cell stimulation, IL-2 gene expression, and T-cell proliferation; these anti-tumor effects have been observed in mouse models of colon carcinoma, metastatic prostate cancer, and metastatic melanoma. PD-1 binds to active T-cells and suppresses T-cell activation; PD-1 antagonists have demonstrated anti-tumor effects as well. CTLA-4 and PD-1 pathway antagonists that may be used in combination with the compounds of general Formula Ia and Formula I disclosed herein include ipilimumab, tremelimumab, nivolumab, pembrolizumab, CT-011, AMP-224, and MDX-1106.

“PD-1 antagonist” or “PD-1 pathway antagonist” means any chemical compound or biological molecule that blocks binding of PD-L1 expressed on a cancer cell to PD-1 expressed on an immune cell (T-cell, B-cell, or NKT-cell) and preferably also blocks binding of PD-L2 expressed on a cancer cell to the immune-cell expressed PD-1. Alternative names or synonyms for PD-1 and its ligands include: PDCD1, PD1, CD279, and SLEB2 for PD-1; PDCDIL1, PDL1, B7H1, B7-4, CD274, and B7-H for PD-L1; and PDCDIL2, PDL2, B7-DC, Btdc, and CD273 for PD-L2. In any of the treatment method, medicaments and uses of the present disclosure in which a human individual is being treated, the PD-1 antagonist blocks binding of human PD-L1 to human PD-1, and preferably blocks binding of both human PD-L1 and PD-L2 to human PD-1. Human PD-1 amino acid sequences can be found in NCBI Locus No.: NP_005009. Human PD-L1 and PD-L2 amino acid sequences can be found in NCBI Locus No.: NP_054862 and NP_079515, respectively.

PD-1 antagonists useful in any of the treatment method, medicaments and uses of the present disclosure include a monoclonal antibody (mAb), or antigen binding fragment thereof, which specifically binds to PD-1 or PD-L1, and preferably specifically binds to human PD-1 or human PD-L1. The mAb may be a human antibody, a humanized antibody, or a chimeric antibody and may include a human constant region. In some embodiments, the human constant region is selected from the group consisting of IgG1, IgG2, IgG3, and IgG4 constant regions, and in preferred embodiments, the human constant region is an IgG1 or IgG4 constant region. In some embodiments, the antigen binding fragment is selected from the group consisting of Fab, Fab′-SH, F(ab′)₂, scFv, and Fv fragments.

Examples of mAbs that bind to human PD-1, and useful in the treatment method, medicaments and uses of the present disclosure, are described in U.S. Pat. Nos. U.S. Pat. Nos. 7,488,802, 7,521,051, 8,008,449, 8,354,509, and 8,168,757, PCT International Patent Application Publication Nos. WO2004/004771, WO2004/072286, and WO2004/056875, and U.S. Patent Application Publication No. US2011/0271358.

Examples of mAbs that bind to human PD-L1, and useful in the treatment method, medicaments and uses of the present disclosure, are described in PCT International Patent Application Nos. WO2013/019906 and WO2010/077634 A1 and in U.S. Pat. No. 8,383,796. Specific anti-human PD-L1 mAbs useful as the PD-1 antagonist in the treatment method, medicaments and uses of the present disclosure include MPDL3280A, BMS-936559, MEDI4736, MSB0010718C, and an antibody that comprises the heavy chain and light chain variable regions of SEQ ID NO:24 and SEQ ID NO:21, respectively, of WO2013/019906.

Other PD-1 antagonists useful in any of the treatment method, medicaments, and uses of the present disclosure include an immune-adhesion that specifically binds to PD-1 or PD-L1, and preferably specifically binds to human PD-1 or human PD-L1, e.g., a fusion protein containing the extracellular or PD-1 binding portion of PD-L1 or PD-L2 fused to a constant region such as an Fc region of an immunoglobulin molecule. Examples of immune-adhesion molecules that specifically bind to PD-1 are described in PCT International Patent Application Publication Nos. WO2010/027827 and WO2011/066342. Specific fusion proteins useful as the PD-1 antagonist in the treatment method, medicaments, and uses of the present disclosure include AMP-224 (also known as B7-DCIg), which is a PD-L2-FC fusion protein and binds to human PD-1.

Thus, the invention further relates to a method of treating cancer in a human patient comprising administration of a compound of Formula Ia and Formula I and a PD-1 antagonist to the patient. The compound of the invention and the PD-1 antagonist may be administered concurrently or sequentially.

In particular embodiments, the PD-1 antagonist is an anti-PD-1 antibody, or antigen binding fragment thereof. In alternative embodiments, the PD-1 antagonist is an anti-PD-L1 antibody, or antigen binding fragment thereof. In some embodiments, the PD-1 antagonist is pembrolizumab (KEYTRUDA™, Merck & Co., Inc., Kenilworth, N.J., USA), nivolumab (OPDIVO™, Bristol-Myers Squibb Company, Princeton, N.J., USA), cemiplimab (LIBTAYO™, Regeneron Pharmaceuticals, Inc., Tarrytown, N.Y., USA), atezolizumab (TECENTRIQ™, Genentech, San Francisco, Calif., USA), durvalumab (IMFINZI™, AstraZeneca Pharmaceuticals LP, Wilmington, Del.), or avelumab (BAVENCIO™, Merck KGaA, Darmstadt, Germany).

In some embodiments, the PD-1 antagonist is pembrolizumab. In particular sub-embodiments, the method comprises administering 200 mg of pembrolizumab to the patient about every three weeks. In other sub-embodiments, the method comprises administering 400 mg of pembrolizumab to the patient about every six weeks.

In further sub-embodiments, the method comprises administering 2 mg/kg of pembrolizumab to the patient about every three weeks. In particular sub-embodiments, the patient is a pediatric patient.

In some embodiments, the PD-1 antagonist is nivolumab. In particular sub-embodiments, the method comprises administering 240 mg of nivolumab to the patient about every two weeks.

In other sub-embodiments, the method comprises administering 480 mg of nivolumab to the patient about every four weeks.

In some embodiments, the PD-1 antagonist is cemiplimab. In particular embodiments, the method comprises administering 350 mg of cemiplimab to the patient about every 3 weeks.

In some embodiments, the PD-1 antagonist is atezolizumab. In particular sub-embodiments, the method comprises administering 1200 mg of atezolizumab to the patient about every three weeks.

In some embodiments, the PD-1 antagonist is durvalumab. In particular sub-embodiments, the method comprises administering 10 mg/kg of durvalumab to the patient about every two weeks.

In some embodiments, the PD-1 antagonist is avelumab. In particular sub-embodiments, the method comprises administering 800 mg of avelumab to the patient about every two weeks.

Examples of other cytotoxic agents include, but are not limited to, arsenic trioxide (sold under the tradename TRISENOX®), asparaginase (also known as L-asparaginase, and Erwinia L-asparaginase, sold under the tradenames ELSPAR® and KIDROLASE®).

Chemotherapeutic agents that may be used in combination with the compounds of general Formula Ia and Formula I disclosed herein include abiraterone acetate, altretamine, anhydrovinblastine, auristatin, bexarotene, bicalutamide, BMS 184476, 2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl)benzene sulfonamide, bleomycin, N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-proly-1-Lproline-tbutylamide, cachectin, cemadotin, chlorambucil, cyclophosphamide, 3′,4′-didehydro-4′deoxy-8′-norvin-caleukoblastine, docetaxol, doxetaxel, cyclophosphamide, carboplatin, carmustine, cisplatin, cryptophycin, cyclophosphamide, cytarabine, dacarbazine (DTIC), dactinomycin, daunorubicin, decitabine dolastatin, doxorubicin (adriamycin), etoposide, 5-fluorouracil, finasteride, flutamide, hydroxyurea and hydroxyureataxanes, ifosfamide, liarozole, lonidamine, lomustine (CCNU), MDV3100, mechlorethamine (nitrogen mustard), melphalan, mivobulin isethionate, rhizoxin, sertenef, streptozocin, mitomycin, methotrexate, taxanes, nilutamide, nivolumab, onapristone, paclitaxel, pembrolizumab, prednimustine, procarbazine, RPR109881, stramustine phosphate, tamoxifen, tasonermin, taxol, tretinoin, vinblastine, vincristine, vindesine sulfate, and vinflunine.

Examples of vascular endothelial growth factor (VEGF) receptor inhibitors include, but are not limited to, bevacizumab (sold under the trademark AVASTIN by Genentech/Roche), axitinib (described in PCT International Patent Publication No. WO01/002369), Brivanib Alaninate ((S)—((R)-1-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-5-methylpyrrolo[2,1-f][1,2,4]triazin-6-yloxy)propan-2-yl)2-aminopropanoate, also known as BMS-582664), motesanib (N-(2,3-dihydro-3,3-dimethyl-TH-indol-6-yl)-2-[(4-pyridinylmethyl)amino]-3-pyridinecarboxamide. and described in PCT International Patent Application Publication No. WO02/068470), pasireotide (also known as SO 230, and described in PCT International Patent Publication No. WO02/010192), and sorafenib (sold under the tradename NEXAVAR).

Examples of topoisomerase II inhibitors, include but are not limited to, etoposide (also known as VP-16 and Etoposide phosphate, sold under the tradenames TOPOSAR, VEPESID, and ETOPOPHOS), and teniposide (also known as VM-26, sold under the tradename VUMON).

Examples of alkylating agents, include but are not limited to, 5-azacytidine (sold under the trade name VIDAZA), decitabine (sold under the trade name of DECOGEN), temozolomide (sold under the trade names TEMODAR and TEMODAL by Schering-Plough/Merck), dactinomycin (also known as actinomycin-D and sold under the tradename COSMEGEN), melphalan (also known as L-PAM, L-sarcolysin, and phenylalanine mustard, sold under the tradename ALKERAN), altretamine (also known as hexamethylmelamine (HMM), sold under the tradename HEXALEN), carmustine (sold under the tradename BCNU), bendamustine (sold under the tradename TREANDA), busulfan (sold under the tradenames BUSULFEX® and MYLERAN®), carboplatin (sold under the tradename PARAPLATIN®), lomustine (also known as CCNU, sold under the tradename CEENU®), cisplatin (also known as CDDP, sold under the tradenames PLATINOL® and PLATINOL®-AQ), chlorambucil (sold under the tradename LEUKERAN®), cyclophosphamide (sold under the tradenames CYTOXAN® and NEOSAR®), dacarbazine (also known as DTIC, DIC and imidazole carboxamide, sold under the tradename DTIC-DOME®), altretamine (also known as hexamethylmelamine (HMM) sold under the tradename HEXALEN®), ifosfamide (sold under the tradename IFEX®), procarbazine (sold under the tradename MATULANE®), mechlorethamine (also known as nitrogen mustard, mustine and mechloroethamine hydrochloride, sold under the tradename MUSTARGEN®), streptozocin (sold under the tradename ZANOSAR®), thiotepa (also known as thiophosphoamide, TESPA and TSPA, and sold under the tradename THIOPLEX®.

Examples of anti-tumor antibiotics include, but are not limited to, doxorubicin (sold under the tradenames ADRIAMYCIN® and RUBEX®), bleomycin (sold under the tradename LENOXANE®), daunorubicin (also known as dauorubicin hydrochloride, daunomycin, and rubidomycin hydrochloride, sold under the tradename CERUBIDINE®), daunorubicin liposomal (daunorubicin citrate liposome, sold under the tradename DAUNOXOME®), mitoxantrone (also known as DHAD, sold under the tradename NOVANTRONE®), epirubicin (sold under the tradename ELLENCE™) idarubicin (sold under the tradenames IDAMYCIN®, IDAMYCIN PFS®), and mitomycin C (sold under the tradename MUTAMYCIN®).

Examples of anti-metabolites include, but are not limited to, claribine (2-chlorodeoxyadenosine, sold under the tradename LEUSTATIN®), 5-fluorouracil (sold under the tradename ADRUCIL®), 6-thioguanine (sold under the tradename PURINETHOL®), pemetrexed (sold under the tradename ALINMTA®), cytarabine (also known as arabinosylcytosine (Ara-C), sold under the tradename CYTOSAR-U®), cytarabine liposomal (also known as Liposomal Ara-C, sold under the tradename DEPOCYT™), decitabine (sold under the tradename DACOGEN®), hydroxyurea (sold under the tradenames HYDREA®, DROXIA™ and MYLOCEL™), fludarabine (sold under the tradename FLUDARA®), floxuridine (sold under the tradename FUDR®), cladribine (also known as 2-chlorodeoxyadenosine (2-CdA) sold under the tradename LEUSTATIN™), methotrexate (also known as amethopterin, methotrexate sodium (MTX), sold under the tradenames RHEUMATREX® and TREXALL™), and pentostatin (sold under the tradename NIPENT®).

Examples of retinoids include, but are not limited to, alitretinoin (sold under the tradename PANRETIN®), tretinoin (all-trans retinoic acid, also known as ATRA, sold under the tradename VESANOID®), isotretinoin (13-c/s-retinoic acid, sold under the tradenames ACCUTANE®, AMNESTEEM®, CLARAVIS®, CLARUS®, DECUTAN®, ISOTANE®, IZOTECH®, ORATANE®, ISOTRET®, and SOTRET®), and bexarotene (sold under the tradename TARGRETIN®).

Further disclosed herein is a compound disclosed herein, or a pharmaceutically acceptable salt thereof, for use in therapy. In one embodiment, disclosed herein is the use of a compound disclosed herein, or a pharmaceutically acceptable salt, solvate or hydrate thereof, for the preparation of a medicament for use in therapy.

Intermediates

Intermediates were either purchased from the following vendors or synthesized using the cited literature references as shown or further modified through additional synthetic manipulations analogous to phosphoramidites described in intermediates I-1 to I-21.

TABLE 1

Intermediates used in the synthesis of Examples of the instant invention.

Structure
Bases
Source

I-1

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B = nucleobase A, C, G, U
ChemGenes Vendor

I-2

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B = nucleobase A, C, G, U
ChemGenes Vendor

I-3

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B = nucleobase A, C, G, U
ChemGenes Vendor

I-4

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B = nucleobase A, C, G, U
ChemGenes Vendor

I-5

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B = nucleobase A, C, G, U
Glen Research Vendor

I-6

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B = nucleobase A, 5-Me-C, G, T
Exiqon Vendor

I-7

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B = nucleobase A, 5-Me-C, G, T
ChemGenes Vendor

I-8

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B = nucleobase A, C, G, U
J . Med. Chem. 2000, 43, 4516-4525

I-9

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B = nucleobase A, C, G, U
Canadian Journal of Chemistry 1989, 67, 831-9

I-11

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B = nucleobase A, C, G, U
ChemGenes Vendor

I-12

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B = nucleobase A, C, G, U
J . Med. Chem. 2006, 49, 1624-1634

I-13

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B = nucleobase A, C, G, U
Nucleic Acids Research, 2009, Vol. 37, No. 4 1353-1362

I-14

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B = nucleobase A, C, G, U
Nuc. Acids Res. 2015, 43, 7189-7200.

I-15

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B = nucleobase A, C, G, U
U.S., 4,849,513, 18 Jul. 1989; Nucleosides & Nucleotides 1992, 11, 1333-51

I-17

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Journal of the Chemical Society, Perkin Transactions 1: Organic and Bio- Organic Chemistry 1995 (12), 1543-50

I-18

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Carbosynth Vendor

I-19

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Carbosynth Vendor

I-20

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Barry and Associates Vendor

I-21

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WO/2017/027646

I-22

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Carbosynth Vendor

I-23

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Journal of Organic Chemistry, 2013, 78, 11271-11282.

I-24

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Barry and Associates Vendor

I-25

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Barry and Associates Vendor

The following experimental procedures detail the preparation of intermediates used in the synthesis of the Examples of the instant invention. The exemplified procedures are for illustrative purposes only, and are not intended to limit the scope of the instant invention in any way.

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Step 1: To a solution of N-(1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)-tetrahydrofuran-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)acetamide (200 g, 701.13 mmol, 1.00 equiv) in pyridine (3000 mL) was added the solution of 1,3-dichloro-1,1,3,3-teraisopropyldisiloxane (TIPSCl₂) (240 g, 762 mmol, 1.1 equiv) in pyridine dropwise. The resulting solution was allowed to react overnight with stirring at room temperature. The reaction mixture was concentrated under vacuum and the residue was extracted with EtOAc three times. The combined organic layer was washed with brine twice and dried over anhydrous Na₂SO₄The filtrate was concentrated under vacuum and the residue was purified by a silica gel column, eluting with dichloromethane/methanol (100:1) to afford of N-(1-((6aR,8R,9R,9aS)-9-hydroxy-2,2,4,4-tetraisopropyl-tetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)acetamide. LCMS-(ES, m/z) found: 528[M+H]⁺. ¹H NMR (300 MHz, Chloroform-d) δ 9.99 (s, 1H), 8.19 (d, J=7.5 Hz, 1H), 7.43 (d, J=7.4 Hz, 1H), 5.80 (s, 1H), 4.32-4.15 (m, 4H), 4.00 (dd, J=13.4, 2.3 Hz, 1H), 3.34 (s, 1H), 2.28 (s, 3H), 1.14-0.86 (m, 28H).

Step 2: To a solution of N-(1-((6aR,8R,9R,9aS)-9-hydroxy-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)acetamide (350 g, 663 mmol, 1.00 equiv) in dichloromethane (1500 mL) was added methyl 1H-1,2,4-triazole-1-carbodithioate (126 g, 794 mmol, 1.20 equiv) in several batches. To the above was added 1,8-diazabicyclo[5.4.0]-undec-7-ene (DBU) (152 g, 996 mmol, 1.50 equiv) dropwise with stirring at 0° C., and stirred overnight at room temperature. The resulting mixture was neutralized with hydrochloric acid (2 M) and the organic layer was washed with sodium bicarbonate solution (sat.), then dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column and eluted with dichloromethane/methanol (100:1) to afford 0-((6aR,8R,9R,9aR)-8-(4-acetamido-2-oxopyrimidin-1(2H)-yl)-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-9-yl) S-methyl carbonodithioate.

LCMS (ES, m/z) found: 618 [M+H]⁺. ¹H NMR (300 MHz, DMSO-d₆) δ 10.91 (s, 1H), 8.05 (d, J=7.5 Hz, 1H), 7.20 (d, J=7.5 Hz, 1H), 6.34 (dd, J=5.3, 1.1 Hz, 1H), 5.80 (d, J=1.1 Hz, 1H), 4.70 (dd, J=8.5, 5.4 Hz, 1H), 4.07-3.91 (m, 3H), 2.58 (s, 3H), 2.09 (s, 3H), 1.07-0.77 (m, 28H).

Step 3: To a solution of 1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (23 g, 80.44 mmol, 5.00 equiv) in dichloromethane (230 mL) was added HF/Py (105 mL, 70% HF in Py) dropwise with stirring at −78° C. The mixture was stirred for 10 minutes at −78° C. To this was added a solution of N-[1-[(6aR,8R,9R,9aR)-9-[[(methylsulfanyl)methanethioyl]oxy]-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-2-oxo-1,2-dihydropyrimidin-4-yl]acetamide (10 g, 16.18 mmol, 1.00 equiv) in dichloromethane (70 mL) dropwise with stirring at −78° C. The resulting solution was stirred for 3 h at 0° C. in an ice/salt bath. The reaction was then quenched by the addition of 100 mL of NaHSO₃(1M). The pH value of the solution was adjusted to 8 with sodium bicarbonate (solid). The solids were filtered out. The resulting solution was extracted with 4×300 mL of ethyl acetate and the combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column with dichloromethane/methanol/TEA (95:5:0.3) as eluent to furnish 800 mg (13%) of N-[1-[(2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)oxolan-2-yl]-2-oxo-1,2-dihydropyrimidin-4-yl]acetamide as a white solid. The above procedure was repeated for ten batches (20 g×10), and the crude was purified together to give N-(1-((2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)tetrahydrofuran-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)acetamide.

LCMS (ES, m/z) found: 354 [M+H]⁺. ¹H NMR (300 MHz, DMSO-d6) δ 10.92 (s, 1H), 8.38 (d, J=7.5 Hz, 1H), 7.18 (d, J=7.5 Hz, 1H), 6.00 (d, J=3.4 Hz, 1H), 5.76 (d, J=6.0 Hz, 1H), 5.28 (t, J=4.9 Hz, 1H), 4.97-4.81 (m, 1H), 4.21 (p, J=5.4 Hz, 1H), 3.92 (dt, J=6.1, 2.9 Hz, 1H), 3.81-3.68 (m, 1H), 3.65-3.52 (m, 1H), 2.07 (s, 3H).

Step 4: To a solution of N-[1-[(2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)oxolan-2-yl]-2-oxo-1,2-dihydropyrimidin-4-yl]acetamide (9 g, 25.48 mmol, 1.00 equiv, co-evaporated with anhydrous pyridine three times) in pyridine (100 mL) was added a solution of 1-[chloro(4-methoxyphenyl)benzyl]-4-methoxybenzene (17.1 g, 50.47 mmol, 2.00 equiv, co-evaporated with anhydrous pyridine three times) in pyridine (200 mL) dropwise with stirring at 30° C. After the resulting solution was stirred for 3 h at 30° C., then concentrated under vacuum. The resulting solution was diluted with 400 mL of DCM, then was washed with 1×300 mL of sodium bicarbonate (sat.) and 1×300 mL of brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column with ethyl acetate/petroleum ether/Et₃N (60:40:0.3) as eluent to furnish N-[1-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl)methoxy]methyl]-4-hydroxy-3-(trifluoromethoxy)oxolan-2-yl]-2-oxo-1,2-dihydropyrimidin-4-yl]acetamide. LCMS (ES, m/z) found: 656 [M+H]⁺. ¹H NMR (400 MHz, Chloroform-d) δ 9.49 (s, 1H), 8.35 (d, J=7.5 Hz, 1H), 7.43 (d, J=7.7 Hz, 2H), 7.39-7.26 (m, 6H), 7.25-7.07 (m, 2H), 6.88 (d, J=8.5 Hz, 4H), 6.16 (s, 1H), 5.16-5.10 (m, 1H), 4.60 (t, J=6.0 Hz, 1H), 4.24 (d, J=7.5 Hz, 1H), 3.83 (s, 6H), 3.65 (d, J=11.3 Hz, 1H), 3.58 (d, J=11.2 Hz, 1H), 3.34 (s, 1H), 2.22 (s, 3H).

Step 5: To a solution of N-[1-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl)methoxy]methyl]-4-hydroxy-3-(trifluoromethoxy)oxolan-2-yl]-2-oxo-1,2-dihydropyrimidin-4-yl]acetamide (13.0 g, 19.83 mmol, 1.00 equiv, which dried by co-evaporation with pyridine twice) in dichloromethane (200 mL) was added DIPEA (12.8 g, 99.04 mmol, 5.00 equiv) dropwise with stirring at 0° C. To the mixture was added 3-([bis(propan-2-yl)amino](chloro)phosphanyloxy)propanenitrile (7.0 g, 29.58 mmol, 1.50 equiv) dropwise with stirring at 0° C. After the resulting solution was stirred for 2 h at room temperature, it then was diluted with 500 mL of CH₂Cl₂. The resulting mixture was washed with 2×300 mL of saturated sodium bicarbonate and 300 mL of brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column with petroleum ether (PE):ethyl acetate (EA):TEA (38:62:0.5) as eluent to obtain the crude. The crude product (13 g) was purified by Flash-Prep-HPLC with the following conditions (IntelFlash-1): Column, C18 silica gel; mobile phase, 50% CH₃CN/H₂O (0.5% NH₄HCO₃) increasing to 80% CH₃CN/H₂O (0.5% NH₄HCO₃) over 30 min; Detector, UV 254 nm. This resulted in N-[1-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl)methoxy]methyl]-4-([[bis(propan-2-yl)amino](2-cyanoethoxy)phosphanyl]oxy)-3-(trifluoromethoxy)oxolan-2-yl]-2-oxo-1,2-dihydropyrimidin-4-yl]acetamide. L-MS (ES, m/z) found: 856 [M+H]⁺. ¹H-NMR (300 MHz, Chloroform-d,ppm): δ 10.69-10.57 (m, 1H), 8.30 (m, 1H), 7.43 (m, 2H), 7.39-7.23 (m, 7H), 7.11 (m, 1H), 6.89 (m, 4H), 6.23 (d, J=2.8 Hz, 1H), 4.98 (m, 1H), 4.66 (m, 1H), 4.40-4.23 (m, 1H), 3.98-3.45 (m, 12H), 2.62 (t, J=6.2 Hz, 1H), 2.43 (m, 1H), 2.28 (d, J=1.8 Hz, 3H), 1.24-1.11 (m, 9H), 1.08 (d, J=6.8 Hz, 3H). F-NMR: (300 MHz, Chloroform-d, ppm): δ 58.148, −58.222. P-NMR: (300 MHz, Chloroform-d, ppm): δ 15.8, 150.9.

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Step 1: I-23C was made in accordance to the procedure described in the procedure for making Intermediate 1-23. To a solution of O-(6aR,8R,9aR)-8-(4-acetamido-2-oxopyrimidin-1(2H)-yl)-2,2,4,4-tetraisopropyl-tetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-9-yl S-methyl carbonodithioate (270 g, 425 mmol, 1.00 equiv) in acetic anhydride (1000 mL) and AcOH (1000 mL) was added sulfuric acid (10 mL) dropwise with stirring at 0° C. under inert atmosphere. The resulting solution was allowed to stir overnight at room temperature. The solvent was concentrated under vacuum and the residue was extracted with EtOAc three times. The combined organic phase was washed with NaHCO₃(sat.) twice and brine once, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column, eluting with dichloromethane/methanol (100:1) and further purification by recrystallization from acetonitrile:ether=1:10 to afford (2R,3R,4R,5R)-5-(4-acetamido-2-oxopyrimidin-1(2H)-yl)-2-(acetoxymethyl)-4 (((methylthio)carbonothioyl)oxy)tetrahydrofuran-3-yl acetate.

LCMS:(ES, m/z) found:460 [M+H]⁺. ¹H NMR (400 MHz, Chloroform-d) δ 10.47 (s, 1H), 7.80 (d, J=7.5 Hz, 1H), 7.50 (d, J=7.6 Hz, 1H), 6.30 (dd, J=5.9, 3.7 Hz, 1H), 6.03 (d, J=3.7 Hz, 1H), 5.53 (t, J=6.1 Hz, 1H), 4.51-4.38 (m, 3H), 2.58 (s, 3H), 2.28 (s, 3H), 2.15 (s, 3H), 2.10 (s, 3H).

Step 2: To a solution of 1,3-dibromo-5-isopropylimidazolidine-2,4-dione (47 g, 165 mmol, 5 equiv) in dichloromethane (300 mL) was added HF/Py (150 mL, 70% HF in Py) dropwise with stirring at −78° C. under inert atmosphere and kept stirring for 10 min. This was followed by the addition of a solution of (2R,3R,4R,5R)-5-(4-acetamido-2-oxopyrimidin-1(2H)-yl)-2-(acetoxymethyl)-4-(((methylthio)carbonothi-oyl)oxy)tetrahydrofuran-3-yl acetate (15 g, 32.68 mmol, 1.00 equiv) in dichloromethane (200 mL) dropwise with stirring at −78° C., and then kept stirring for 2 h at −20° C. The resulting mixture was quenched by the addition of NaHSO₃(sat.) and extracted with dichloromethane four times. The combined organic layers was washed with sodium bicarbonate (sat.) twice and brine once, and dried over anhydrous sodium sulfate. The filtrate was concentrated and residue was purified by recrystallization from ether to afford (2R,3R,4R,5R)-5-(4-acetamido-2-oxopyrimidin-1(2H)-yl)-2-(acetoxymethyl)-4-(trifluoromethoxy)tetrahydrofuran-3-yl acetate. LCMS (ES, m/z) found: 438 [M+H]⁺. ¹H NMR (300 MHz, Chloroform-d) δ 10.13 (s, 1H), 7.91 (d, J=7.6 Hz, 1H), 7.46 (d, J=7.6 Hz, 1H), 5.91 (d, J=1.5 Hz, 1H), 5.16 (d, J=5.9 Hz, 2H), 4.57-4.17 (m, 3H), 2.24 (s, 3H), 2.16-1.97 (m, 6H).

Step 3: To a solution of 2R,3R,4R,5R)-5-(4-acetamido-2-oxopyrimidin-1(2H)-yl)-2-(acetoxymethyl)-4-(trifluoromethoxy)tetrahydrofuran-3-yl acetate (38 g, 87 mmol, 1.00 equiv) in MeOH (260 mL) was added NH₃(130 mL, 7 M in MeOH) dropwise at 0° C. The resulting solution was stirred overnight at room temperature and then concentrated under vacuum to give 4-amino-1-((2R, 3R, 4R, 5R)-4-hydroxy-5-(hydro-xylmethyl)-3-(trifluoromethoxy)-tetrahydrofuran-2-yl) pyrimidin-2(1H)-one. LCMS: (ES, m/z): 312 [M+H]⁺.

Step 4: To a solution of crude 4-amino-1-((2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluorometh-oxy)-tetrahydrofuran-2-yl)pyrimidin-2(1H)-one (15 g, 48 mmol, 1.00 equiv) in water (100 mL) was added NaNO₂(67 g, 971 mmol, 20 equiv) in several batches, and followed added acetic acid (50 mL) dropwise at 0° C. The resulting solution was stirred overnight at room temperature and concentrated under vacuum. The residue was triturated with EtOH and the filtrate was concentrated under vacuum after filtration to afford 1-((2R, 3R, 4R, 5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)-tetrahydrofuran-2-yl) pyrimidine-2,4(1H, 3H)-dione. LCMS (ES, m/z): 313[M+H]⁺.

Step 5: To a solution of 1-((2R, 3R, 4R, 5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)-tetrahydrofuran-2-yl) pyrimidine-2, 4(1H, 3H)-dione (14 g, 45 mmol, 1.00 equiv, coevaporate with pyridine twice previously) in pyridine (250 mL) was added a solution of 1-(chlorobis(4-methoxyphenyl) methyl)benzene (23 g, 67 mmol, 1.50 equiv) in pyridine (50 mL) dropwise with stirring at room temperature under inert atmosphere. The resulting solution was stirred overnight at room temperature and then concentrated under vacuum. The residue was extracted with EtOAc twice and the combined organic layer was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column, eluting with dichloromethane/methanol/Et₃N (100:1:0.5) to afford 1-((2R, 3R, 4R, 5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-4-hydroxy-3-(trifluoromethoxy)-tetrahydrofuran-2-yl) pyrimidine-2, 4(1H, 3H)-dione. ¹H NMR (300 MHz, Chloroform-d) δ 8.75 (d, J=2.0 Hz, 1H), 7.68 (d, J=8.2 Hz, 1H), 7.36-7.17 (m, 9H), 6.90-6.77 (m, 4H), 6.21 (d, J=5.2 Hz, 1H), 5.30 (dd, J=8.2, 2.2 Hz, 1H), 4.91 (t, J=5.0 Hz, 1H), 4.53 (q, J=4.5 Hz, 1H), 4.18-4.11 (m, 2H), 3.77 (s, 6H), 3.54-3.50 (m, 1H), 2.50 (d, J=5.0 Hz, 1H).

Step 6: 1-((2R, 3R, 4R, 5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-4-hydroxy-3-(trifluoro-methoxy)-tetrahydrofuran-2-yl) pyrimidine-2, 4(1H, 3H)-dione (20 g, 33 mmol, 1.00 equiv) was co-evaporated with dry pyridine three times and then diluted with dry dichloromethane (200 mL), To the above solution was added N-ethyl-N-isopropylpropan-2-amine (13 g, 99 mmol, 3.00 equiv), followed by addition of 3-chloro-3-(diisopropylamino) phosphinooxypropanenitrile (12 g, 50 mmol, 1.50 equiv) dropwise with stirring at room temperature under inert atmosphere. The resulting solution was stirred for 2 h at room temperature and then quenched with sodium bicarbonate (sat.) and extracted with dichloromethane twice. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column, eluting with ethyl acetate/hexane/EtN₃(50:100:0.75) and then further purified by a reverse column, eluting with acetonitrile: water (containing 0.1% NH₄HCO₃) from 20:80 to 80:20 over 1.5 h to afford (2R,3R,4R,5R)-2-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-(trifluoromethoxy)tetrahydrofuran-3-yl (2-cyanoethyl) diisopropylphosphoramidite. LCMS (ES, m/z) found: 815 [M+H]⁺. ¹H-NMR (400 MHz, CDCl₃, ppm): 8.82 (s, 1H), 7.76 (dd, J=11.9, 8.1 Hz, 1H), 7.42-7.25 (m, 7H), 6.88 (dd, J=8.6, 3.8 Hz, 4H), 6.23 (dd, J=9.3, 4.9 Hz, 1H), 5.33 (dd, J=8.3, 4.2 Hz, 1H), 5.01 (q, J=5.7 Hz, 1H), 4.69 (ddt, J=14.7, 9.4, 4.7 Hz, 1H), 4.34 (d, J=3.5 Hz, 1H), 4.23 (d, J=4.5 Hz, 0H), 3.93 (dt, J=13.4, 6.4 Hz, 0H), 3.86-3.72 (m, 6H), 3.65 (dq, J=12.3, 6.3, 5.8 Hz, 3H), 3.61-3.44 (m, 2H), 2.66 (t, J=6.2 Hz, 1H), 2.48 (s, 1H), 1.21 (t, J=5.5 Hz, 10H), 1.10 (d, J=6.7 Hz, 2H). ³¹P NMR-(400 MHz, CDCl₃, ppm): 151.5, 152.0. ¹⁹F NMR: (400 MHz, CDCl₃, ppm): δ−58.6, −59.1.

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Step 1: To a solution of 1H-1,2,4-triazole (250 g, 3.62 mol, 1.00 equiv) in acetone (7000 mL) was added K₃PO₄(768 g, 3.62 mol, 1.00 equiv). After the mixture was stirred for 20 minutes at 25° C., then was added CS₂(826 g, 3.00 equiv) dropwise with stirring at 25° C. After the mixture was stirred for 20 minutes at 25° C., then was added CH₃I (515 g, 3.63 mol, 1.00 equiv) dropwise with stirring at 25° C. After the resulting solution was stirred overnight at 25° C., the solids were filtered out. The filtrate was concentrated under vacuum, then was dissolved in 5 L of ethyl acetate. The insoluble solids were filtered out. The filtrate was concentrated under vacuum to get the crude. The crude product was re-crystallized from EA:Et₂O:PE with the ratio of 1:1:1 to afford 1-(methylsulfanyl)carbothioyl-1H-1,2,4-triazole. LCMS (ES, m/z) found: 160[M+H]⁺. ¹H NMR (300 MHz, Chloroform-d) δ 9.14 (s, 1H), 8.06 (s, 1H), 2.72 (s, 3H).

Step 2: To a solution of N-[9-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-9H-purin-6-yl]benzamide (200 g, 538.58 mmol, 1.00 equiv) in anhydrous pyridine (2500 mL) was added (dichloro-4-sulfanylidene)(titanio)phosphane (186 g, 1.02 mol, 1.11 equiv) dropwise with stirring at room temperature. After the resulting solution was stirred overnight at room temperature, the crude mixture was concentrated under vacuum. The residue was dissolved in 1000 mL of dichloromethane, then was washed with 2×400 mL of sodium bicarbonate (sat.), dried over anhydrous sodium sulfate. The residue was purified by a silica gel column with ethyl acetate/petroleum ether (2:1) as eluent to N-[9-[(6aR,8R,9R,9aS)-9-hydroxy-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-9H-purin-6-yl]benzamide. LCMS (ES, m/z) found: 614 [M+H]⁺. ¹H NMR (300 MHz, DMSO-d₆) δ 11.17 (s, 1H), 8.63 (s, 1H), 8.50 (s, 1H), 8.05-7.96 (m, 2H), 7.61 (t, J=7.3 Hz, 1H), 7.52 (dd, J=8.3, 6.7 Hz, 2H), 5.97 (d, J=1.4 Hz, 1H), 5.64 (d, J=4.7 Hz, 1H), 4.79 (dd, J=8.3, 5.1 Hz, 1H), 4.62 (t, J=5.3 Hz, 1H), 4.10-3.94 (m, 2H), 3.97-3.86 (m, 1H), 1.07-0.88 (m, 28H).

Step 3: To a solution of N-[9-[(6aR,8R,9R,9aS)-9-hydroxy-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-9H-purin-6-yl]benzamide (246 g, 400.75 mmol, 1.00 equiv) in dichloromethane (600 mL) was added 1-(methylsulfanyl)carbothioyl-1H-1,2,4-triazole (115 g, 722.21 mmol, 1.80 equiv). This was followed by the addition of DBU (80 g, 525.49 mmol, 1.30 equiv) dropwise with stirring at 25° C. After the resulting solution was stirred for overnight at 25° C., then was washed with 2×300 mL of hydrogen chloride (1 M) and 1×300 mL of sodium bicarbonate (sat.), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column with ethyl acetate/petroleum ether (1:1) as eluent to furnish N-[9-[(6aR,8R,9R,9aR)-9-[[(methylsulfanyl)methanethioyl]oxy]-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-9H-purin-6-yl]benzamide. LCMS (ES, m/z) found: 704[M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆) δ 11.28 (s, 1H), 8.64 (d, J=18.1 Hz, 2H), 8.09-8.02 (m, 2H), 7.67 (t, J=7.4 Hz, 1H), 7.57 (t, J=7.6 Hz, 2H), 6.77 (dd, J=5.7, 1.4 Hz, 1H), 6.41 (d, J=1.4 Hz, 1H), 5.45 (dd, J=8.4, 5.8 Hz, 1H), 4.05 (ddd, J=14.2, 11.1, 6.9 Hz, 3H), 2.65 (s, 3H), 1.28-0.74 (m, 28H).

Step 4: To a solution of 1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (20.5 g, 71.70 mmol, 5.00 equiv) in dichloromethane (150 mL) was added a solution of 70% HF in Py (60 mL) dropwise with stirring at −78° C. The mixture was stirred for 10 minutes at −78° C., then added a solution of N-[9-[(6aR,8R,9R,9aR)-9-[[(methylsulfanyl)methanethioyl]oxy]-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-9H-purin-6-yl]benzamide (10 g, 14.20 mmol, 1.00 equiv) in dichloromethane (50 mL) dropwise with stirring at −78° C. The mixture was stirred for 10 minutes at −78° C. followed by stirring for 3 h at −5° C. The reaction was then quenched by the addition of 10 g of NaHSO₃. The pH value of the solution was adjusted to 7-8 with sodium bicarbonate (sat.). The solids were filtered out. The filtrate was extracted with 2×200 mL of dichloromethane and the combined organic layers were dried over anhydrous sodium sulfate. The residue was purified by a silica gel column with dichloromethane/methanol (25:1) as eluent to furnish N-[9-[(2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)oxolan-2-yl]-9H-purin-6-yl]benzamide. LCMS (ES, m/z) found: 440 [M+H]⁺. ¹H-NMR: (300 MHz, CDCl₃, ppm): δ 8.81 (s, 1H), 8.14-7.98 (m, 3H), 7.70-7.48 (m, 3H), 6.14 (d, J=7.4 Hz, 1H), 5.62 (dd, J=7.5, 4.4 Hz, 1H), 4.72 (d, J=4.4 Hz, 1H), 4.40 (s, 1H), 3.99 (dd, J=13.2, 1.6 Hz, 1H), 3.84-3.68 (m, 1H), 3.48 (s, 2H).

Step 5: N-[9-[(2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)oxolan-2-yl]-9H-purin-6-yl]benzamide (11 g, 25.04 mmol, 1.00 equiv) was co-evaporated with anhydrous pyridine three times before added into a 500-mL 3-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen. This was followed by the addition of anhydrous pyridine (110 mL) and a solution of 1-[chloro(4-methoxyphenyl)benzyl]-4-methoxybenzene (17 g, 50.17 mmol, 2.00 equiv, was dried by co-evaporation with anhydrous pyridine three times) in anhydrous pyridine (170 mL) dropwise with stirring at 30° C. The resulting solution was stirred for 3 h at 30° C., then was concentrated under vacuum. The resulting solution was diluted with 500 mL of dichloromethane, then was washed with 2×200 mL of sodium bicarbonate (sat.) and 1×200 mL of brine (sat.), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column with PE:EA:TEA (50:50:0.3) as eluent to furnish N-[9-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl)methoxy]methyl]-4-hydroxy-3-(trifluoromethoxy)oxolan-2-yl]-9H-purin-6-yl]benzamide. LCMS (ES, m/z) found: 742 [M+H]⁺.

¹H NMR (400 MHz, Chloroform-d) δ 9.06 (s, 1H), 8.69 (s, 1H), 8.13 (s, 1H), 8.04 (d, J=8.0 Hz, 2H), 7.63 (t, J=7.3 Hz, 1H), 7.54 (t, J=7.7 Hz, 2H), 7.46-7.39 (m, 2H), 7.35-7.19 (m, 5H), 7.19 (d, J=8.6 Hz, 1H), 6.84 (t, J=9.1 Hz, 4H), 6.33 (d, J=5.9 Hz, 1H), 5.83 (t, J=5.4 Hz, 1H), 4.75 (dd, J=10.1, 4.9 Hz, 1H), 4.32 (q, J=3.6 Hz, 1H), 3.81 (d, J=8.2 Hz, 6H), 3.58 (dd, J=10.7, 3.6 Hz, 1H), 3.44 (dd, J=10.8, 3.8 Hz, 1H), 3.13-3.03 (m, 1H).

Step 6: N-[9-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl)methoxy]methyl]-4-hydroxy-3-(trifluoromethoxy)oxolan-2-yl]-9H-purin-6-yl]benzamide (13.5 g, 18.20 mmol, 1.00 equiv) was co-evaporated with anhydrous pyridine three times before added into a 250-mL 3-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen. This was followed by the addition of dichloromethane (140 mL) and DIPEA (12 g, 92.85 mmol, 5.00 equiv). This was followed by the addition of 3-([bis(propan-2-yl)amino](chloro)phosphanyloxy)-propanenitrile (6.5 g, 27.5 mmol, 1.50 equiv) dropwise with stirring at 20° C. The resulting solution was stirred for 1 h at 20° C., then quenched by 5 mL of methanol. The resulting solution was diluted with 500 mL of EA, then was washed with 1×200 mL of sodium bicarbonate (sat.) and 1×200 mL of brine (sat.), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column with PE:EA:TEA (50:50:0.3) as eluent to furnish a crude. The crude was purified by a C18 silica gel column with H₂O(0.5% NH₄HCO₃):ACN(1:99). This resulted in N-[9-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl)methoxy]methyl]-4-([[bis(propan-2-yl)amino](2-cyanoethoxy)phosphanyl]oxy)-3-(trifluoromethoxy)oxolan-2-yl]-9H-purin-6-yl]benzamide. LCMS (ES, m/z) found: 942 [M+H]⁺. ¹H-NMR: (300 MHz, CDCl₃, ppm): δ 8.97 (d, J=4.9 Hz, 1H), 8.65 (d, J=1.6 Hz, 1H), 8.12 (d, J=4.3 Hz, 1H), 8.04-7.94 (m, 2H), 7.67-7.44 (m, 3H), 7.39 (dt, J=8.0, 1.7 Hz, 2H), 7.32-7.10 (m, 7H), 6.84-6.71 (m, 4H), 6.26 (dd, J=16.3, 6.1 Hz, 1H), 5.90-5.78 (m, 1H), 4.79 (dddd, J=17.7, 12.7, 7.2, 3.7 Hz, 1H), 4.34 (q, J=3.5 Hz, 1H), 3.86 (ddd, J=12.3, 6.0, 1.4 Hz, 1H), 3.75 (d, J=2.4 Hz, 6H), 3.60 (dddd, J=20.4, 13.7, 8.5, 3.9 Hz, 4H), 3.34 (ddd, J=10.7, 6.9, 3.8 Hz, 1H), 2.60 (t, J=6.3 Hz, 1H), 2.39 (td, J=6.4, 2.1 Hz, 1H), 1.19 (dd, J=7.0, 5.7 Hz, 8H), 1.06 (d, J=6.7 Hz, 4H). ³¹PNMR (400 MHz, CDCl₃, ppm): 152.1, 151.9. ¹⁹FNMR (400 MHz, CDCl₃, ppm): δ−59.0, 59.2.

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Step 1: To a solution of 2-amino-9-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6,9-dihydro-1H-purin-6-one (100 g, 353.06 mmol, 1.00 equiv) in N,N-dimethylformamide (1000 mL) was added imidazole (88.8 g, 1.31 mol, 3.70 equiv). This was followed by the addition of TiPSCl₂(166.9 g, 529.84 mmol, 1.50 equiv) dropwise with stirring at room temperature. After the resulting solution was stirred for 2.5 h at room temperature, then quenched by the addition of 1500 mL of water/ice. The solids were collected by filtration and washed with water, then dried in an oven at 50° C. This resulted in (crude) of 9-[(6aR,8R,9R,9aS)-9-hydroxy-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo[3,2-f] [1,3, 5,2,4] trioxadisilicon-8-yl]-2-amino-6,9-dihydro-1H-purin-6-one. LC-MS (ES, m/z) found: 526 [M+H]^{+ 1}H NMR (300 MHz, DMSO-d6, ppm) δ 10.61 (s, 1H), 7.73 (s, 1H), 6.45 (s, 2H), 5.64 (d, J=1.8 Hz, 1H), 5.56 (s, 1H), 4.37-4.18 (m, 2H), 4.11-3.83 (m, 3H), 1.05-0.86 (m, 28H).

Step 2: To a solution of 9-[(6aR,8R,9R,9aS)-9-hydroxy-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo [3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-2-amino-6,9-dihydro-1H-purin-6-one (182 g, crude, 346.17 mmol, 1.00 equiv) in dichloromethane (1500 mL), was added 1-(methylsulfanyl)carbothioyl-1H-1,2,4-triazole (71.5 g, 449.03 mmol, 1.20 equiv) under atmosphere of argon. To this was added DBU (79 g, 518.92 mmol, 1.50 equiv) dropwise with stirring at room temperature. After the resulting solution was stirred for 3 h at room temperature, then washed with 2×1000 mL of brine (sat.) and concentrated under vacuum. The residue was purified by a silica gel column with dichloromethane/methanol (30:1) as eluent to furnish 9-[(6aR,8R,9R,9aR)-9-[[(methylsulfanyl)methanethioyl]oxy]-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4] trioxadisilicon-8-yl]-2-amino-6,9-dihydro-TH-purin-6-one. LC-MS (ES, m/z) found: 616 [M+H]⁺. ¹H NMR (300 MHz, DMSO-d6) δ 10.69 (s, 1H), 7.88 (s, 1H), 6.38 (dd, J=5.9, 2.7 Hz, 1H), 6.36 (s, 2H), 5.98 (d, J=2.6 Hz, 1H), 4.85-4.74 (m, 1H), 3.99 (tdt, J=12.3, 8.7, 4.1 Hz, 3H), 2.56 (s, 3H), 1.14-0.73 (m, 28H).

Step 3: To a solution of 9-[(6aR,8R,9R,9aR)-9-[[(methylsulfanyl)methanethioyl]oxy]-2,2,4,4-tetrakis (propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-2-amino-6,9-dihydro-1H-purin-6-one (113 g, 183.47 mmol, 1.00 equiv) in pyridine (1200 mL), was added TMSCl (100 g, 920.47 mmol, 5.00 equiv) dropwise with stirring at 0° C. and stirred for 2 hours at room temperature. To this was added i-Bu₂O (49.3 g, 312.03 mmol, 1.70 equiv) dropwise with stirring at 0° C. After the resulting solution was stirred for 1 h at room temperature, it then was quenched by the addition of 150 mL saturated aqueous of sodium bicarbonate. The pyridine was evaporated out under vacuum, then diluted with 700 mL of brine (sat.), and extracted with 2 x 800 mL of ethyl acetate. The combined organic layers were concentrated and purified by a silica gel column with dichloromethane/methanol (100% DCM) as eluent to furnish N-[9-[(6aR,8R,9R,9aR)-9-[[(methylsulfanyl)methanethioyl]oxy]-2,2,4,4-tetrakis (propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-6-oxo-6,9-dihydro-1H-purin-2-yl]-2-methylpropanamide. LCMS (ES, m/z) found: 686 [M+H]⁺

Step 4: To a solution of N-[9-[(6aR,8R,9R,9aR)-9-[[(methylsulfanyl)methanethioyl]oxy]-2,2,4,4-tetrakis (propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-6-oxo-6,9-dihydro-TH-purin-2-yl]-2-methylpropanamide (90 g, 131.19 mmol, 1.00 equiv) in acetic anhydride (420 mL) and HOAc (210 mL), was added sulfuric acid (conc.) (25.2 g, 256.93 mmol, 1.96 equiv) dropwise with stirring at 0° C. After the resulting solution was stirred overnight at room temperature, then diluted with 700 mL of brine (sat.). The pH value of the solution was adjusted to 7 with a solution of sodium carbonate (4 M), then extracted with dichloromethane 4×1000 ml and the combined organic layer were concentrated under vacuum. The residue was purified by a silica gel column with ethyl acetate/petroleum ether (up to 100% EA) as eluent to furnish [(2R,3R,4R,5R)-3-(acetyloxy)-5-[2-(2-methylpropanamido)-6-oxo-6,9-dihydro-1H-purin-9-yl]-4-[[(methylsulfanyl) methane thioyl]oxy] oxolan-2-yl]methyl acetate. LCMS (ES, m/z) found: 528 [M+H]⁺

Step 5: To a solution of 1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (27.1 g, 94.78 mmol, 5.00 equiv) in dichloromethane (140 mL), was added a solution of 70% HF in Py (100 mL) dropwise with stirring at −78° C. To this was added [(2R,3R,4R,5R)-3-(acetyloxy)-5-[2-(2-methylpropanamido)-6-oxo-6,9-dihydro-1H-purin-9-yl]-4-[[(methylsulfanyl)methanethioyl]oxy]oxolan-2-yl]methyl acetate (10 g, 18.95 mmol, 1.00 equiv) with stirring at −78° C. After the resulting solution was stirred for 1 h at −78° C., then quenched by the addition of 100 mL saturated aqueous of NaHSO₃. The resulting solution was extracted with 3×200 mL of ethyl acetate and the organic layers combined. The pH value of the organic layers was adjusted to 8 with a saturated solution of sodium bicarbonate. Then the aqueous phase was extracted with ethyl acetate, and the combined organic layers were concentrated under vacuum. The residue was purified by a silica gel column with ethyl acetate/petroleum ether (up to 100% EA) as eluent to furnish [(2R,3R,4R,5R)-3-(acetyloxy)-5-[2-(2-methylpropanamido)-6-oxo-6,9-dihydro-1H-purin-9-yl]-4-(trifluoromethoxy)oxolan-2-yl]methyl acetate. LCMS (ES, m/z) found: 506 [M+H]⁺.

Step 6: To a solution of guanidine nitrate (11.81 g, 96.84 mmol, 5 equiv), MeONa (1.88 g, 34.86 mmol, 1.80 equiv) in dichloromethane (100 mL) and methanol (900 mL), was added [(2R,3R,4R,5R)-3-(acetyloxy)-5-[2-(2-methylpropanamido)-6-oxo-6,9-dihydro-1H-purin-9-yl]-4-(trifluoromethoxy)oxolan-2-yl]methyl acetate (9.8 g, 19.39 mmol, 1.00 equiv) with stirring at −5° C. After the resulting solution was stirred for 30 min at −5° C., the pH was adjusted 7 with citric acid (1 M) and concentrated under vacuum. The residue was purified by a silica gel column with dichloromethane/methanol (10:1) as eluent to furnish N-[9-[(2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)oxolan-2-yl]-6-oxo-6,9-dihydro-1H-purin-2-yl]-2-methylpropanamide. LCMS (ES, m/z) found: 422 [M+H]⁺.

Step 7: To a solution of N-[9-[(2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)oxolan-2-yl]-6-oxo-6,9-dihydro-1H-purin-2-yl]-2-methylpropanamide (7.15 g, 16.97 mmol, 1.00 equiv) in pyridine (70 mL), was added 1-methoxy-4-[1-(4-methoxyphenyl)-1-phenylethyl]benzene (8.63 g, 27.10 mmol, 1.50 equiv) at room temperature. After the resulting solution was stirred for 1.5 h at room temperature, it was concentrated under vacuum. The residue was purified by a silica gel column with EA/PE (up to 80% EA) as eluent to furnish N-[9-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl) methoxy] methyl]-4-hydroxy-3-(trifluoromethoxy)oxolan-2-yl]-6-oxo-6,9-dihydro-1H-purin-2-yl]-2-methylprop anamide. LCMS: (ES, m/z) found: 724 [M+H]⁺.

Step 8: To a solution of N-[9-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl)methoxy]methyl]-4-hydroxy-3-(trifluoromethoxy)oxolan-2-yl]-6-oxo-6,9-dihydro-1H-purin-2-yl]-2-methylpropanamide (11.1 g, 15.34 mmol, 1.00 equiv) in dichloromethane (110 mL), was added DIPEA (9.9 g, 76.74 mmol, 5.00 equiv) with stirring at room temperature. To this was added 3-([[bis(propan-2-yl)amino](chloro) phosphanyl]oxy)propanenitrile (7.25 g, 30.63 mmol, 2.00 equiv) dropwise with stirring at room temperature. After the solution was stirred for 1.5 h at room temperature, it was then quenched by the addition of 20 mL of methanol. The resulting mixture was washed with 2×150 mL saturated solution of sodium bicarbonate, and the organic layers concentrated under vacuum. The crude product was purified by Flash-Prep-HPLC with the following conditions (CombiFlash-1): C18 column; mobile phase, 5% ACN/NH₄CO₃0.05% in H₂O increasing to 80% ACN/NH₄CO₃0.05% in H₂O within 35 min; Detector, UV 254 nm. This resulted in N-[9-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl) (phenyl) methoxy]methyl]-4-([[bis(propan-2-yl)amino](2-cyanoethoxy)phosphanyl]oxy)-3-(trifluoro meth oxy) oxolan-2-yl]-6-oxo-6,9-dihydro-1H-purin-2-yl]-2-methylpropanamide as a white solid. LCMS: (ES, m/z): 924 [M+H]⁺. ¹H-NMR: (300 MHz, DMSO, ppm): δ 12.15 (s, 1H), 11.52 (s, 1H), 8.24 (d, J=1.4 Hz, 1H), 7.42-7.16 (m, 9H), 6.84 (m, 4H), 6.17 (dd, J=18.6, 6.2 Hz, 1H), 5.78 (m, 1H), 4.62-4.45 (m, 1H), 4.26 (m, 1H), 3.74 (d, J=2.0 Hz, 7H), 3.69-3.39 (m, 3H), 3.41 (m, 1H), 3.25 (dd, J=10.7, 3.0 Hz, 1H), 2.83-2.68 (m, 2H), 2.59 (t, J=5.9 Hz, 1H), 1.14 (m, 14H), 1.01 (d, J=6.7 Hz, 4H). ¹⁹FNMR: (300 MHz, DMSO, ppm): δ−57.612, −57.740. ³¹PNMR: (300 MHz, DMSO, ppm):150.775, 150.450.

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Step 1: To a slurry of (3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-3-methyldihydrofuran-2(3H)-one (WO2013/188480, 2013, A1) (50 g, 308 mmol) in DCM (500 mL) was added pyridine (64.8 mL). The mixture was stirred in an ice bath and then benzoyl chloride (95 g, 678 mmol) was added dropwise over 65 minutes at −5° C. The reaction mixture was then warmed to room temperature and stirred for 1.5 h. The reaction mixture was then cooled with an ice bath and water (215 mL) was added. The reaction mixture was warmed to room temperature and stirred for 2 h at this temperature. The organic layer was washed with aqueous hydrogen chloride (1 N, 170 mL), water (170 mL), saturated aqueous sodium carbonate (170 mL), water (3×170 mL) and brine (170 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was treated with toluene (100 mL) and it began to solidify. The solid was filtered and washed with toluene (100 mL) to give ((2R,3R,4R)-3-(benzoyloxy)-4-hydroxy-4-methyl-5-oxotetrahydrofuran-2-yl)methyl benzoate. MS ESI calculated for C20H1807 [M+H]⁺ 371.11, found 371.10. ¹H-NMR: (300 MHz, DMSO-d₆) δ 8.10-7.97 (m, 2H), 7.96-7.84 (m, 2H), 7.72-7.59 (m, 2H), 7.58-7.41 (m, 4H), 6.40 (s, 1H), 5.42 (d, J=7.1 Hz, 1H), 4.98 (td, J=6.4, 3.7 Hz, 1H), 4.67 (dd, J=12.3, 3.7 Hz, 1H), 4.59 (dd, J=12.4, 6.0 Hz, 1H), 1.45 (s, 3H).

Step 2: To a solution of ((2R,3R,4R)-3-(benzoyloxy)-4-hydroxy-4-methyl-5-oxotetrahydrofuran-2-yl)methyl benzoate (49 g, 132 mmol) in DCM (353 mL) was added dropwise diethylaminosulfurtrifluoride (34.1 g, 212 mmol) over 1 h at −78° C. The reaction was slowly warmed to room temperature and stirred for 16 h. The reaction was then quenched with saturated aqueous sodium carbonate (150 mL). The organic layer was washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to give ((2R,3R,4S)-3-(benzoyloxy)-4-fluoro-4-methyl-5-oxotetrahydrofuran-2-yl)methyl benzoate. MS ESI calculated for C₂₀H₁₇FO₆[M+H]⁺ 373.10, found 373.10. ¹H-NMR: (400 MHz, CDCl₃) δ 8.10-8.02 (m, 4H), 7.69-7.63 (m, 1H), 7.63-7.56 (m, 1H), 7.54-7.48 (m, 2H), 7.47-7.41 (m, 2H), 5.87 (dd, J=18.8, 5.3 Hz, 1H), 4.83-4.72 (m, 2H), 4.73-4.62 (m, 1H), 1.79 (d, J=23.2 Hz, 3H). ¹⁹F-NMR: (400 MHz, CDCl₃) δ−153.28 (s, 1F).

Step 3: To a solution of ((2R,3R,4S)-3-(benzoyloxy)-4-fluoro-4-methyl-5-oxotetrahydrofuran-2-yl)methyl benzoate (62 g, 167 mmol) in dry THF (500 mL) was added dropwise lithium tri-tert-butoxyaluminum hydride (1M in THF, 200 mL, 200 mmol) at −78° C. within 30 min. The reaction was warmed to room temperature and stirred for 2 h at this temperature. The mixture was cooled to 0° C. and quenched by saturated aqueous ammonium chloride (800 mL), diluted with ethyl acetate (1000 mL). The resulting mixture was then filtered and the organic layer was washed with brine (800 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give ((2R,3R,4S)-3-(benzoyloxy)-4-fluoro-5-hydroxy-4-methyltetrahydrofuran-2-yl)methyl benzoate which was used directly in next step. MS ESI calculated for C₂₀H₁₉FO₆[M+H]⁺ 375.12, found 357.10. ¹⁹F-NMR: (282 MHz, CDCl₃) δ−149.67, −167.30.

Step 4: To a solution of ((2R,3R,4S)-3-(benzoyloxy)-4-fluoro-5-hydroxy-4-methyltetrahydrofuran-2-yl)methyl benzoate (67.3 g, 180 mmol) (co-evaporated with dry toluene (100 mL×3) before being used) in dry DCM (565 mL) were added triethylamine (32.5 mL, 234 mmol), N,N-dimethylpyridin-4-amine (4.39 g, 36.0 mmol) and acetic anhydride (25.5 mL, 270 mmol) at 0° C. The reaction mixture was then warmed to room temperature and stirred for 2 h at this temperature. The solvent was removed under reduced pressure and the residue was purified by column chromatography on silica gel using a stepwise gradient of ethyl acetate (0-20%) in petroleum ether to give (2R,3R,4S)-5-acetoxy-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate. MS ESI calculated for C₂₂H₂₁FO₇[M+NH₄]⁺ 434.13, found 434.20.

Step 5: To a solution of (2R,3R,4S)-5-acetoxy-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate (80 g, 192 mmol) in DCM (720 mL) was added hydrogen bromide (33% in acetic acid, 95 mL, 576 mmol) at 0° C. The reaction was stirred at ambient temperature for 16 h. The reaction mixture was quenched by sodium bicarbonate until no carbon dioxide was observed and diluted with DCM (800 mL). The organic layer was then washed with saturated aqueous sodium bicarbonate (3×700 mL), water (700 mL), brine (700 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give ((2R,3R,4S)-3-(benzoyloxy)-5-bromo-4-fluoro-4-methyltetrahydrofuran-2-yl)methyl benzoate which was used directly in next step. MS ESI calculated for C₂₀H₁₈BrFO₅[M+H]⁺ 437.03, 439.03, found 437.15, 439.05.

Step 6: To a solution of N-(9H-purin-6-yl)benzamide (45.2 g, 189 mmol) in anhydrous dioxane (700 mL) was added sodium hydride (60% in mineral oil, 9.35 g, 234 mmol) at room temperature under argon atmosphere and the mixture was stirred for 5 min at this temperature. Then a solution of ((2R,3R,4S)-3-(benzoyloxy)-5-bromo-4-fluoro-4-methyltetrahydrofuran-2-yl)methyl benzoate (78.6 g, 180 mmol) in anhydrous dioxane (700 mL) was added at room temperature. The reaction mixture was stirred at 100° C. for 5 h. The reaction mixture was quenched by saturated aqueous ammonium chloride (800 mL) and diluted with ethyl acetate (1000 mL). The organic layer was then washed with water (800 mL), brine (800 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel using a stepwise gradient of 0-60% ethyl acetate in petroleum ether to give a crude product. The crude product was further purified by Combi-Flash with the following conditions: Column: AQ-C18 gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 45-100% within 35 min); Flow rate: 100 mL/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and evaporated under reduced pressure to give (2R,3R,4S,5R)-5-(6-benzamido-9H-purin-9-yl)-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate.

MS ESI calculated for C₃₂H₂₆FN₅O₆[M+H]⁺ 596.19, found 596.20. ¹H-NMR: (400 MHz, CDCl₃) δ 9.12 (s, 1H), 8.84 (s, 1H), 8.37 (d, J=3.3 Hz, 1H), 8.18-8.09 (m, 4H), 8.09-8.01 (m, 2H), 7.74-7.42 (m, 9H), 6.52 (d, J=20.4 Hz, 1H), 5.68 (dd, J=16.8, 2.6 Hz, 1H), 4.90 (dd, J=12.0, 5.9 Hz, 1H), 4.84 (dd, J=12.0, 4.1 Hz, 1H), 4.60-4.57 (m, 1H), 1.61 (d, J=22.6 Hz, 3H). ¹⁹F-NMR: (376 MHz, CDCl₃) δ−160.17 (s, 1F).

Step 7: To a solution of (2R,3R,4S,5R)-5-(6-benzamido-9H-purin-9-yl)-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate (10 g, 16.79 mmol) in methanol (70 mL), THF (56 mL) and water (14 mL) was added dropwise aqueous sodium hydroxide solution (4 N, 12.59 mL, 50.4 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 25 min. The reaction mixture was quenched by formic acid (1.89 mL). Then the solvent was removed under reduced pressure. The crude product was purified by column chromatography on silica gel using a stepwise gradient of MeOH (0-10%) in DCM to give N-(9-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-9H-purin-6-yl)benzamide. MS ESI calculated for C₁₈H₁₈FN₅O₄[M+H]⁺ 388.13, found 388.15. ¹H-NMR: (400 MHz, CD₃OD): δ 8.76 (s, 1H), 8.64 (d, J=2.7 Hz, 1H), 8.13-8.06 (m, 2H), 7.71-7.62 (m, 1H), 7.62-7.53 (m, 2H), 6.37 (d, J=16.3 Hz, 1H), 4.33 (dd, J=19.1, 4.5 Hz, 1H), 4.03 (q, J=4.8 Hz, 1H), 3.92 (dd, J=12.1, 4.2 Hz, 1H), 3.87 (dd, J=12.1, 5.5 Hz, 1H), 1.51 (d, J=23.0 Hz, 3H). ¹⁹F-NMR: (376 MHz, CD₃OD): δ−159.16 (s, 1F).

Step 8: To a solution of N-(9-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-9H-purin-6-yl)benzamide (8.8 g, 22.72 mmol) in dry pyridine (45 mL) was added 4,4′-(chloro(phenyl)methylene)bis(methoxybenzene) (8.47 g, 24.99 mmol) at room temperature. The reaction mixture was stirred at room temperature for 2 h. The solvent was removed under reduced pressure. The residue was co-evaporated with toluene (3×40 mL) and then purified by column chromatography on silica gel using a stepwise gradient of ethyl acetate (0-100%) in petroleum ether to give a yellow crude product. The crude product was further purified by Combi-Flash with the following conditions: Column: AQ-C18 gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 45-100% within 35 min); Flow rate: 100 mL/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and roto-evaporated in vacuo to give N-(9-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)-9H-purin-6-yl)benzamide. MS ESI calculated for C₃₉H₃₆FN₅O₆[M+H]⁺ 690.26, found 690.25.

¹H-NMR: (300 MHz, CD₃OD) δ 8.69 (s, 1H), 8.33 (d, J=2.9 Hz, 1H), 8.10-8.01 (m, 2H), 7.65-7.43 (m, 5H), 7.36-7.15 (m, 7H), 6.82 (d, J=8.9 Hz, 4H), 6.33 (d, J=17.0 Hz, 1H), 4.29 (dd, J=18.8, 4.3 Hz, 1H), 4.14-4.10 (m, 1H), 3.74 (s, 6H), 3.46 (dd, J=10.1, 6.4 Hz, 1H), 3.39 (dd, J=10.1, 4.0 Hz, 1H), 1.43 (d, J=23.1 Hz, 3H).

Step 9: To a solution of 3-((bis(diisopropylamino)phosphino)oxy)propanenitrile (8.46 g, 28.1 mmol) (co-evaporated with dry acetonitrile (3×20 mL) before being used) in dry acetonitrile (20 mL) was added pyridin-1-ium 2,2,2-trifluoroacetate (5.42 g, 28.1 mmol) at room temperature. The solution was marked as solution A. To a solution of N-(9-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)-9H-purin-6-yl)benzamide (12.9 g, 18.70 mmol) (co-evaporated with dry acetonitrile (3×50 mL) before being used) in dry acetonitrile (45 mL) was added to solution A at room temperature. The reaction was stirred at room temperature for 1 h. The reaction was quenched by water (300 mL) and diluted with ethyl acetate (400 mL). The organic layer was washed with saturated aqueous sodium bicarbonate (300 mL), water (300 mL) and brine (300 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified by Combi-Flash with the following conditions: Column: AQ-C18 gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 45-100% within 35 min); Flow rate: 100 m/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and the solvent was removed under reduced pressure to give (2R,3R,4S,5R)-5-(6-benzamido-9H-purin-9-yl)-2-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl (2-cyanoethyl) diisopropylphosphoramidite. MS ESI calculated for C₄₈H₅₃FN₇O₇P [M+H]⁺ 890.37, found 890.45. ¹H-NMR: (400 MHz, DMSO-d₆) δ 11.30 (s, 1H), 8.76 (s, 0.6H), 8.75 (s, 0.4H), 8.44 (d, J=2.3 Hz, 0.4H), 8.40 (d, J=2.4 Hz, 0.6H), 8.11-8.04 (m, 2H), 7.71-7.62 (m, 1H), 7.61-7.53 (m, 2H), 7.52-7.40 (m, 2H), 7.35-7.20 (m, 7H), 6.90-6.85 (m, 4H), 6.39 (d, J=16.0 Hz, 0.4H), 6.38 (d, J=16.0 Hz, 0.6H), 4.76-4.61 (m, 1H), 4.31-4.19 (m, 1H), 3.82-3.70 (m, 1H), 3.74 (s, 6H), 3.66-3.43 (m, 5H), 2.75 (t, J=5.8 Hz, 1H), 2.59 (t, J=5.9 Hz, 1H), 1.59 (d, J=22.9 Hz, 1.5H), 1.53 (d, J=23.0 Hz, 1.5H), 1.23-0.96 (m, 12H). ¹⁹F-NMR: (376 MHz, DMSO-d₆) δ−154.29 (d, J=3.7 Hz, 0.5F), −154.91 (s, 0.5F). ³¹P-NMR: (162 MHz, DMSO-d₆) δ 150.40 (d, J=3.7 Hz, 0.5P), 150.14 (s, 0.5P).

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Step 1: To a suspension of N-(2-oxo-1,2-dihydropyrimidin-4-yl)benzamide (88 g, 408 mmol) in dry acetonitrile (200 mL) was added (E)-trimethylsilyl N-(trimethylsilyl)acetimidate (225 mL, 919 mmol) dropwise at room temperature under an argon atmosphere. The mixture was then heated to 70° C. and stirred for 2 h. The reaction was cooled to room temperature and a solution of (2R,3R,4S)-5-acetoxy-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate (85 g, 204 mmol) (co-evaporated with dry acetonitrile (3×50 mL) before being used) in dry acetonitrile (225 mL) was added at room temperature. The mixture was heated to 70° C. and stirred for 10 h. The reaction was diluted with ethyl acetate (1500 mL) and quenched by saturated aqueous sodium bicarbonate (600 mL) at 0° C. and the mixture was filtered. The organic layer was washed with saturated sodium bicarbonate (3×600 mL), water (600 mL) and brine (800 mL). The organic layer was dried with anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel using a stepwise gradient of ethyl acetate (0-50%) in petroleum ether to give (2R,3R,4S,5R)-5-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate. MS ESI calculated for C₃₁H₂₆FN₃O₇[M+H]⁺ 572.18, found 572.15. ¹H-NMR: (400 MHz, DMSO-d₆) δ 11.36 (s, 1H), 8.18-7.99 (m, 7H), 7.81-7.48 (m, 9H), 7.41 (d, J=7.6 Hz, 1H), 6.42 (d, J=17.6 Hz, 1H), 5.60 (dd, J=19.7, 4.6 Hz, 1H), 4.80-4.74 (m, 2H), 4.68-4.61 (m, 1H), 1.60 (d, J=23.5 Hz, 3H). ¹⁹F-NMR: (376 MHz, DMSO) δ−158.48 (s, 1F).

Step 2: To a solution of (2R,3R,4S,5R)-5-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate (14.1 g, 24.67 mmol) in THF (99 mL), MeOH (79 mL) and water (19.7 mL) was added dropwise aqueous sodium hydroxide (4 N, 18.50 mL, 74.0 mmol) at 0° C. The reaction was then stirred at room temperature for 20 min. The reaction was quenched by formic acid (2.82 mL, 74.0 mmol) at 0° C. The solvent was then removed under reduced pressure and the residue was purified by column chromatography on silica gel using a stepwise gradient of MeOH (0-10%) in DCM to give N-(1-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)benzamide. MS ESI calculated for C17H₁₈FN₃O₅[M+H]⁺ 364.12, found 364.15. ¹H-NMR: (400 MHz, DMSO-d₆) δ 11.31 (s, 1H), 8.23 (d, J=7.6 Hz, 1H), 8.06-7.97 (m, 2H), 7.69-7.60 (m, 1H), 7.62-7.49 (m, 2H), 7.41-7.36 (m, 1H), 6.09 (d, J=13.8 Hz, 1H), 5.93 (d, J=5.7 Hz, 1H), 5.12 (t, J=5.6 Hz, 1H), 4.13-4.05 (m, 1H), 3.82-3.78 (m, 1H), 3.77-3.72 (m, 1H), 3.68-3.62 (m, 1H), 1.46 (d, J=23.6 Hz, 3H). ¹⁹F-NMR: (376 MHz, DMSO-d₆) δ−156.84 (s, 1F).

Step 3: To a solution of N-(1-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)benzamide (7.2 g, 19.82 mmol) (co-evaporated with dry pyridine (3×20 mL) before being used) in dry pyridine (36 mL) was added 4,4′-(chloro(phenyl)methylene)bis(methoxybenzene) (7.39 g, 21.80 mmol) at room temperature. The reaction was stirred at room temperature for 2 h. The solvent was removed under reduced pressure and the residue was co-evaporated with toluene (2×15 mL). The residue was purified by column chromatography on silica gel using a stepwise gradient of ethyl acetate (0-100%) in petroleum ether to give a crude product. The crude product was further purified by Combi-Flash with the following conditions: Column: AQ C18 gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 45-100% within 35 min); Flow rate: 100 mL/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and the solvent was removed under reduced pressure to give N-(1-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)benzamide. MS ESI calculated for C₃₈H₃₆FN₃O₇[M+H]⁺ 666.25, found 666.25. ¹H-NMR: (400 MHz, DMSO-d₆) δ 11.32 (s, 1H), 8.05-8.01 (m, 3H), 7.69-7.60 (m, 1H), 7.58-7.49 (m, 2H), 7.44 (d, J=7.8 Hz, 2H), 7.38-7.26 (m, 8H), 6.97-6.90 (m, 4H), 6.13 (d, J=13.2 Hz, 1H), 6.03 (d, J=5.7 Hz, 1H), 4.16 (dt, J=21.4, 6.0 Hz, 1H), 4.01-3.97 (m, 1H), 3.77 (s, 6H), 3.42-3.33 (m, 2H), 1.46 (d, J=23.6 Hz, 3H). ¹⁹F-NMR: (376 MHz, DMSO-d₆) δ−156.48 (s, 1F).

Step 4: To a solution of 3-(((diisopropylamino)(2,4-dimethylpentan-3-yl)phosphino)oxy)propanenitrile (6.50 g, 21.63 mmol) (co-evaporated with dry acetonitrile (3×20 mL) before being used) in dry acetonitrile (20 mL) was added pyridine 2,2,2-trifluoroacetate (4.18 g, 21.63 mmol) at room temperature. After 5 min, a solution of N-(1-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)benzamide (9.6 g, 14.42 mmol) (co-evaporated with dry acetonitrile (3×20 mL) before being used) in dry acetonitrile (30 mL) was added at room temperature. The reaction was stirred at room temperature for 1.5 h. The reaction was diluted with ethyl acetate (500 mL) and quenched by saturated aqueous sodium bicarbonate (300 mL). The organic layer was then washed with water (300 mL) and brine (300 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was further purified by Combi-Flash with the following conditions: Column: AQ C18 gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 45-100% within 35 min); Flow rate: 100 mL/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and roto-evaporated in vacuo to give (2R,3R,4S,5R)-5-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-2-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl (2-cyanoethyl) diisopropylphosphoramidite. MS ESI calculated for C₄₇H₅₃FN₅O₈P [M+H]⁺ 866.36, found 866.45. ¹H-NMR: (400 MHz, DMSO-d₆) δ 11.35 (s, 1H), 8.12 (d, J=7.9 Hz, 0.5H), 8.10 (d, J=8.0 Hz, 0.5H), 8.05-7.98 (m, 2H), 7.69-7.60 (m, 1H), 7.58-7.49 (m, 2H), 7.49-7.39 (m, 2H), 7.38-7.26 (m, 8H), 6.97-6.88 (m, 4H), 6.23-6.12 (m, 1H), 4.51-4.41 (m, 1H), 4.16-4.11 (m, 1H), 3.77 (s, 6H), 3.59-3.40 (m, 6H), 2.75 (t, J=5.9 Hz, 1H), 2.59 (t, J=6.0 Hz, 1H), 1.58 (d, J=14.1 Hz, 1.4H), 1.52 (d, J=14.0 Hz, 1.6H), 1.22-0.93 (m, 12H). ¹⁹F-NMR: (376 MHz, DMSO-d₆) δ−155.59 (s, 0.5F), −155.99 (s, 0.5F). ³¹P-NMR: (162 MHz, DMSO-d₆) δ 150.64 (s, 0.5P), 150.46 (s, 0.5P).

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Step 1: To a solution of uracil (16.9 g, 144 mmol) in acetonitrile (150 mL) was added N,O-bis(trimethylsilyl)acetamide (84 mL, 324 mmol) at 85° C. The solution was stirred at 85° C. for 2 h and then cooled to room temperature. A solution of (2R,3R,4S)-5-acetoxy-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate (30.0 g, 72 mmol) in acetonitrile (150 mL) was added followed by dropwise addition of trimethylsilyl trifluoromethanesulfonate (84 g, 360 mmol) at room temperature. The reaction mixture was slowly warmed to 85° C. and stirred for 16 h. The mixture was then cooled to 0° C., quenched with saturated sodium bicarbonate solution (500 mL). The mixture was diluted with ethyl acetate (500 mL). The separated organic layer was washed with brine (3×100 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with 10%-50% ethyl acetate in petroleum ether to afford ((2R,3R,4S)-3-(benzoyloxy)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-4-methyltetrahydrofuran-2-yl)methyl benzoate. MS ESI calculated for C₂₄H₂₁FN₂O₇[M+H]⁺ 469.13, found 469.15. ¹H-NMR (300 MHz, CDCl₃) δ 9.48 (brs, 0.5H), 9.40 (brs, 0.5H), 8.21-7.87 (m, 5H), 7.76-7.36 (m, 6H), 6.56 (d, J=15.5 Hz, 0.5H), 6.30 (d, J=20.3 Hz, 0.5H), 5.86-5.70 (m, 1H), 5.62-5.45 (m, 1H), 5.00-4.57 (m, 3H), 1.66 (d, J=11.4, 1.5H), 1.56 (d, J=11.4 Hz, 1.5H).

Step 2: To a solution of ((2R,3R,4S)-3-(benzoyloxy)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-4-methyltetrahydrofuran-2-yl)methyl benzoate (48 g, 103 mmol) in MeOH (125 mL), THF (100 mL) and H₂O (25 mL) was added aqueous sodium hydroxide solution (77.8 mL, 4 M, 309 mmol) at 0° C. The mixture was stirred at room temperature for 30 min. The pH of the mixture was adjusted to 7 with formic acid and the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with 0%-20% MeOH in DCM to afford 1-((3S, 4R, 5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, 3H)-dione as a white solid. The product (containing α and β isomers) was separated by Prep-SFC with the following conditions: (Column: ChiralPak IA-SFC, 5 cm×25 cm, 5 um; Mobile Phase A: CO₂: 50, Mobile Phase B: MeOH: 50; Flow rate:180 mL/min; 220 nm; RT1: 2.51 min (β isomer); RT2: 3.46 min (a isomer)) to give 1-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)-dione. MS ESI calculated for C₁₀H₁₃FN₂O₅[M+H]⁺ 261.08, found 261.10. ¹H-NMR (300 MHz, CD₃OD) δ 7.82 (dd, J=8.2, 2.4 Hz, 1H), 6.03 (d, J=15.9 Hz, 1H), 5.70 (d, J=8.2 Hz, 1H), 4.13 (dd, J=20.7, 4.7 Hz, 1H), 3.92-3.79 (m, 2H), 3.84-3.68 (m, 1H), 1.48 (d, J=23.3 Hz, 3H).

Step 3: To a solution of 1-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)-dione (9.5 g, 365.0 mmol) in pyridine (60 mL) was added 4,4′-(chloro(phenyl)methylene)bis(methoxybenzene) (13.6 g, 402.0 mmol) at 0° C. The mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure. The residue was purified by Combi Flash Column Chromatography as the following conditions: (Column: AQ-C¹⁸, 19×250 mm, 10 um; Mobile Phase A: Water, Mobile Phase B: MeCN; Flow rate: 100 mL/min; Gradient: 30% B hold 5 min, 30% B to 100% B in 35 min; 100% B hold 8 min, 254/280 nm). The product-containing fractions were pooled and concentrated under reduced pressure to afford 1-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)-dione. MS ESI calculated for C₃₁H₃₁FN₂O₇[M−H]⁻561.21, found 561.21. ¹H-NMR (400 MHz, CD₃OD) δ 7.78-7.70 (m, 1H), 7.50-7.43 (m, 2H), 7.39-7.19 (m, 7H), 6.93-6.82 (m, 4H), 6.03 (d, J=14.1 Hz, 1H), 5.47 (dd, J=8.0, 2.1 Hz, 1H), 4.27 (dd, J=20.7, 5.6 Hz, 1H), 3.96 (q, J=4.7 Hz, 1H), 3.83-3.74 (m, 6H), 3.44 (d, J=4.2 Hz, 2H), 1.48 (d, J=23.3 Hz, 3H).

Step 4: 1-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)-dione (15.0 g, 26.6 mmol) was co-evaporated with dry acetonitrile (3×10 mL) and then re-dissolved in DCM (40 mL), marked as solution A which was kept under an argon atmosphere before being used. 3-(Bis(diisopropylamino)phosphinooxy)propanenitrile (12.0 g, 39.9 mmol) was also co-evaporated with dry acetonitrile (3×10 mL) and then re-dissolved in DCM (35 mL), marked as solution B. 4,5-Dicyanoimidazole (2.6 g, 22.0 mmol) was added into the solution B, followed by the addition of solution A at ambient temperature. The resulting mixture was stirred at ambient temperature for 2 h. The reaction mixture was diluted with DCM (300 mL) and washed with saturated aqueous sodium bicarbonate solution (300 mL) and the organic layer was concentrated under reduced pressure. The residue was purified by Combi Flash Column Chromatography as the following conditions: (Column: AQ-C¹⁸, 19×250 mm, 10 um; Mobile Phase A: Water, Mobile Phase B: MeCN; Flow rate: 100 mL/min; Gradient: 30% B hold 5 min, 30% B to 100% B in 30 min; 100% B hold 8 min, 254/280 nm). The product-containing fractions were pooled and concentrated under reduced pressure to afford (2R,3R,4S,5R)-2-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-4-methyltetrahydrofuran-3-yl (2-cyanoethyl) diisopropylphosphoramidite. MS ESI calculated for C₄₀H₄₈FN₄O₈P [M+H]⁺ 763.32, found 763.30. ¹H-NMR (300 MHz, DMSO-d₆) δ 11.55 (s, 1H), 7.63-7.60 (m, 1H), 7.46-7.42 (m, 2H), 7.37-7.24 (m, 7H), 6.94-6.85 (m, 4H), 6.05-5.95 (m, 1H), 5.60-5.46 (m, 1H), 4.47-4.33 (m, 1H), 4.15-4.03 (m, 1H), 3.85-3.65 (m, 1H), 3.76 (s, 6H), 3.65-3.29 (m, 5H), 2.75 (t, J=5.8 Hz, 1H), 2.58 (t, J=5.9 Hz, 1H), 1.55-1.43 (m, 3H), 1.14-0.98 (m, 12H). ¹⁹F-NMR (282 MHz, DMSO-d₆): δ−155.85 (s, 0.3F), −156.32 (s, 0.7F). ³¹P-NMR (121 MHz, DMSO-d₆): δ 150.57 (s, 0.4P), 150.23 (s, 0.6P).

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Step 1: To a suspension of ((2R,3R,4S)-3-(benzoyloxy)-4-fluoro-5-hydroxy-4-methyltetrahydrofuran-2-yl)methyl benzoate (74 g, 198 mmol), 6-chloro-9H-purin-2-amine (36.9 g, 217 mmol) and triphenylphosphine (156 g, 593 mmol) in dry THF (740 mL) was added (E)-diethyl diazene-1,2-dicarboxylate (93 mL, 593 mmol) at 0° C. The reaction was then stirred at ambient temperature for 3 h. After completion of the reaction, the solvent was removed and the residue was diluted with ethyl acetate (1000 mL). The organic layer was then washed with saturated aqueous sodium bicarbonate solution (2×500 mL), water (500 mL) and brine (500 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was then purified by column chromatography on silica gel using a stepwise gradient of ethyl acetate (0-50%) in petroleum ether to give product which is a mixture of α and β isomers. The crude product was used as it. MS ESI calculated for C₄₃H₃₄ClFN₅O₅P [M+H]⁺ 786.20, 788.20, found 786.25, 788.25.

Step 2: To a suspension of ((2R,3R,4S,5R)-3-(benzoyloxy)-5-(6-chloro-2-((triphenylphosphoranylidene)amino)-9H-purin-9-yl)-4-fluoro-4-methyltetrahydrofuran-2-yl)methyl benzoate (133 g, 169 mmol) in EtOH (700 mL) was added acetic acid (700 mL, 1 M in water, 700 mmol) at ambient temperature. The reaction was then stirred at 80° C. for 2 h. After completion of the reaction, the solvent was removed and the residue was diluted with ethyl acetate (1000 mL) and neutralized with sodium bicarbonate (200 g). The organic layer was then washed with saturated aqueous sodium bicarbonate solution (6×500 mL), water (500 mL) and brine (500 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel using a stepwise gradient of ethyl acetate (0-50%) in petroleum ether to give a crude product. To the crude product was then added Et₂O (300 mL). The resulting solid, which contains triphenylphosphine oxide and most of the a isomer, was filtered off and the filtrate was concentrated under reduced pressure. The residue was then purified by column chromatography on silica gel using a stepwise gradient of ethyl acetate (0-50%) in petroleum ether to give (2R,3R,4S,5R)-5-(2-amino-6-chloro-9H-purin-9-yl)-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate. MS ESI calculated for C₂₅H₂₁ClFN₅O₅[M+H]⁺ 526.12, 528.12, found 526.20, 528.20. ¹H-NMR: (300 MHz, DMSO-d₆) δ 8.14 (d, J=3.2 Hz, 1H), 8.13 (m, 4H), 7.79-7.51 (m, 6H), 7.12 (brs, 2H), 6.25 (d, J=18.8 Hz, 1H), 5.71 (dd, J=18.9, 4.0 Hz, 1H), 4.83-4.69 (m, 2H), 4.69-4.65 (m, 1H), 1.55 (d, J=23.1 Hz, 3H). ¹⁹F-NMR: (282 MHz, DMSO-d₆) δ−157.00 (d, J=1.9 Hz, 1F).

Step 3: To (2R,3R,4S,5R)-5-(2-amino-6-chloro-9H-purin-9-yl)-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate (22 g, 41.8 mmol) was added ammonia (200 mL, 7 M in 2-propanol, 1400 mmol) at −78° C. The reaction was then stirred at 90° C. for 2 d. After completion of the reaction, the solvent was removed and ethyl acetate (150 mL) was added. The suspension was filtered and the filter cake was washed with ethyl acetate (30 mL) to give (2R,3R,4S,5R)-5-(2,6-diamino-9H-purin-9-yl)-4-fluoro-2-(hydroxymethyl)-4-methyltetrahydrofuran-3-ol. MS ESI calculated for C₁₁H₁₅FN₆O₃[M+H]⁺ 299.12, found 299.10.

¹H-NMR: (300 MHz, DMSO-d₆): δ 7.77 (d, J=3.1 Hz, 1H), 6.80 (brs, 2H), 6.02 (s, 1H), 5.88 (brs, 2H), 5.86 (d, J=17.9 Hz, 1H), 5.07 (s, 1H), 4.15 (d, J=19.3 Hz, 1H), 3.84-3.73 (m, 1H), 3.73-3.55 (m, 2H), 1.36 (d, J=23.1 Hz, 3H). ¹⁹F-NMR: (282 MHz, DMSO-d₆): δ−157.21 (s, 1F).

Step 4: To a solution of (2R,3R,4S,5R)-5-(2,6-diamino-9H-purin-9-yl)-4-fluoro-2-(hydroxymethyl)-4-methyltetrahydrofuran-3-ol (11.5 g, 38.6 mmol) in Tris-buffer (193 mL, pH=7.5) was added adenosine deaminase (193 mg, 38.6 mmol) at ambient temperature. The mixture was then warmed to 40° C. and stirred for 16 h. The mixture was concentrated. The residue was purified by Combi-Flash Column Chromatography with the following conditions: Column: AQ C18gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 0-20% within 50 min); Flow rate: 100 mL/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and the solvent was removed under reduced pressure to give 2-amino-9-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-1H-purin-6(9H)-one. MS ESI calculated for C₁₁H₁₄FN₅O₄[M+H]⁺ 300.10, found 300.10. ¹H-NMR: (300 MHz, DMSO-d₆): δ 7.58 (d, J=3.1 Hz, 1H), 7.01 (brs, 2H), 6.16 (brs, 1H), 5.77 (d, J=17.6 Hz, 1H), 5.16 (brs, 1H), 4.11 (dd, J=19.3, 4.5 Hz, 1H), 3.79-3.66 (m, 1H), 3.62-3.54 (m, 2H), 1.31 (d, J=23.0 Hz, 3H). ¹⁹F-NMR: (282 MHz, DMSO-d₆): δ−157.34 (s, 1F).

Step 5: To a suspension of 2-amino-9-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-1H-purin-6(9H)-one (6.2 g, 20.72 mmol) in dry pyridine (31 mL) was added trimethylsilyl chloride (9.6 mL, 103.7 mmol) at 0° C. The reaction was then warmed to room temperature and continued to stir for 2 h. Then isobutryic anhydride (3.2 mL, 31.1 mmol) was added at 0° C. The mixture was stirred at room temperature for 1.5 h. After completion of the reaction, ammonium hydroxide (100 mL) was added. The solvents were removed under reduced pressure and the residue was purified by chromatography on silica gel using a stepwise gradient of MeOH (0-20%) in DCM to give N-(9-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-6-oxo-6,9-dihydro-1H-purin-2-yl)isobutyramide. MS ESI calculated for C₁₅H₂₀FN₅O₅[M+H]⁺ 370.14, found 370.10.

¹H-NMR: (400 MHz, DMSO-d₆) δ 12.13 (s, 1H), 11.67 (s, 1H), 8.09 (d, J=2.7 Hz, 1H), 6.06 (d, J=5.1 Hz, 1H), 5.91 (d, J=15.8 Hz, 1H), 5.10 (t, J=5.7 Hz, 1H), 4.20 (dt, J=18.5, 4.9 Hz, 1H), 3.88-3.79 (m, 1H), 3.76-3.57 (m, 2H), 2.78 (h, J=6.8 Hz, 1H), 1.40 (d, J=23.2 Hz, 3H), 1.13 (d, J=6.8 Hz, 6H). ¹⁹F-NMR: (282 MHz, DMSO-d₆): δ−157.74 (s, 1F).

Step 6: To a solution of N-(9-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-6-oxo-6,9-dihydro-1H-purin-2-yl)isobutyramide (3 g, 8.12 mmol) in dry pyridine (15 mL) was added 4,4′-(chloro(phenyl)methylene)bis(methoxybenzene) (3.02 g, 8.9 mmol) at room temperature. The reaction was quenched by adding MeOH (5 mL). The solvent was removed and the residue was purified by Combi-Flash Colum Chromatography with the following conditions: Column: AQ C18gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 60-100% within 50 min); Flow rate: 100 mL/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and the solvent was removed under reduced pressure to give N-(9-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)-6-oxo-6,9-dihydro-1H-purin-2-yl)isobutyramide. MS ESI calculated for C₃₆H₃₈FN₅O₇[M+H]⁺ 672.28, found 672.25. ¹H-NMR: (400 MHz, DMSO-d₆) δ 12.15 (s, 1H), 11.68 (s, 1H), 7.86 (d, J=2.9 Hz, 1H), 7.48-7.38 (m, 2H), 7.35-7.18 (m, 7H), 6.93-6.81 (m, 4H), 6.09 (d, J=5.0 Hz, 1H), 5.97 (d, J=16.8 Hz, 1H), 4.20 (dt, J=18.8, 4.8 Hz, 1H), 4.08-3.98 (m, 1H), 3.74 (s, 6H), 3.44-3.29 (m, 1H), 3.23 (dd, J=10.3, 3.6 Hz, 1H), 2.78 (septet, J=6.8 Hz, 1H), 1.39 (d, J=23.2 Hz, 3H), 1.13 (dd, J=6.8, 1.7 Hz, 6H). ¹⁹F-NMR: (376 MHz, DMSO-d₆): δ−157.52 (s, 1F).

Step 7: To a solution of 3-((bis(diisopropylamino)phosphino)oxy)propanenitrile (11.30 g, 37.5 mmol)(co-evaporated with dry MeCN (3×60 mL) before being used) in dry DCM (24 mL) was added 4,5-dicyanoimidazole (4.4 g, 37.5 mmol) at room temperature. The solution was marked as solution A. To a solution of N-(9-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)-6-oxo-6,9-dihydro-1H-purin-2-yl)isobutyramide (16.8 g, 25.01 mmol) (co-evaporated with dry MeCN (3×60 mL) before being used) in dry DCM (60 mL) was added solution A at room temperature under argon. The reaction was then stirred at room temperature for 1 h. After completion of the reaction, the reaction mixture was diluted with DCM (300 mL). The organic layer was then washed with saturated aqueous sodium bicarbonate solution (150 mL), water (150 mL) and brine (150 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was then purified by Combi-Flash Column Chromatography with the following conditions: Column: AQ C18 gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 45-100% within 45 min); Flow rate: 100 mL/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and roto-evaporated in vacuo to give (2R,3R,4S,5R)-2-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-4-fluoro-5-(2-isobutyramido-6-oxo-1H-purin-9(6H)-yl)-4-methyltetrahydrofuran-3-yl (2-cyanoethyl) diisopropylphosphoramidite. MS ESI calculated for C₄₅H₅₅FN₇O₈P [M+H]⁺ 872.38, found 872.40. ¹H-NMR: (400 MHz, DMSO-d₆) δ 12.19 (s, 1H), 11.69 (s, 0.4H), 11.66 (s, 0.6H), 7.97-7.90 (m, 1H), 7.48-7.39 (m, 2H), 7.37-7.20 (m, 7H), 6.96-6.85 (m, 4H), 6.04-5.94 (m, 1H), 4.58-4.38 (m, 1H), 4.27-4.12 (m, 1H), 3.86-3.66 (m, 7H), 3.66-3.46 (m, 3H), 3.45-3.27 (m, 2H), 2.87-2.72 (m, 2H), 2.64-2.55 (m, 1H), 1.58-1.41 (m, 3H), 1.18-1.09 (m, 16H), 1.00 (d, J=6.7 Hz, 2H). ¹⁹F-NMR: (376 MHz, DMSO-d₆): δ−155.28 (s, 0.4F), −155.50 (s, 0.6F). ³¹P-NMR: (162 MHz, DMSO-d₆) δ 150.98 (s, 0.4P), 150.03 (s, 0.6P).

TABLE 2

Exemplary nomenclature for first position nucleotides starting at the 5′ end include:

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rGs

Wherein L is selected from the Table 2a:

Also included within Table 2 above are first position nucleotides where “r” in “rG” or “rGs” represent groups comprising “ome”, “flu,” “thior,” “d,” “fara,” “1na,” “moe,” “4alkd,” “ara,” “meF” “cf3,” 25, rxylo, ami, ome4S, fara8az, and “me,” as illustrated herein.

TABLE 2a

L Definition

5′ end

substituents L
Structure

HO

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pcp

ppp

An embodiment of this invention is realized when the first position nucleotide starting at the 5′ end of Formula Ia and Formula I is referred to as rG wherein V, Y, Z, W, and X are O, —OH, —CH—, N, and O, respectively. An embodiment of this invention is realized when the first position nucleotide starting at the 5′ end of Formula Ia and Formula I is referred to as rGs wherein V, Y, Z, W, and X are O, —SH, —CH—, N, and O, respectively.

Table 3: Nomenclature for internally-located nucleotides (i.e., positions 2-23) contained within the single-stranded molecules exemplified in Table 5. For the purposes of this invention, in Table 3 the terms “r,” “ome”, “flu,” “thior,” “d,” “fara,” “1na,” “moe,” “4alkd,” “ara,” “meF” “cf3,” 25, rxylo, ami, ome4S, fara8az, or “me,” appearing before the “B” in the first and third column of Table 3 represents a sugar group as exemplified in columns two and four of Table 3, and “B” represents a nucleobase or derivative thereof selected from adenine, cytosine, guanine and uracil, which are designated “A”, “C”, “G”, and “U”, respectively.

TABLE 3

Internal

nucleotide

nomenclature

(positions 2-23)
Structure

rB B = nucleobase A, C, G, U, or I where “I” in “rI” is an “A” analog

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omeB B = nucleobase A, C, G, U

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fluB B = nucleobase A, C, G, U

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thiorB B = nucleobase A, C, G, U

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dB B = nucleobase A, C, G, U

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faraB B = nucleobase A, C, G, U

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lnaB B = nucleobase A, 5-Me-C, G,5- Me-U

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moeB B = nucleobase A, 5-Me-C, G,5- Me-U

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4alkdB B = nucleobase A, C, G, U

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araB B = nucleobase A, C, G, U

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meFB B = nucleobase A, C, G, U

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cf3B B = nucleobase A, C, G, U

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25B B = nucleobase A, C, G, U

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rxyloB B = nucleobase A, C, G, U

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amiB B = nucleobase A, C, G, U

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ome4SB B = nucleobase A, C, G, U

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rBs B = nucleobase A, C, G, U, or I where “I” in “rI” is an “A” analog

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omeBs B = nucleobase A, C, G, U

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fluBs B = nucleobase A, C, G, U

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thiorBs B = nucleobase A, C, G, U

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dBs B = nucleobase A, C, G, U

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faraBs B = nucleobase A, C, G, U

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lnaBs B = nucleobase A, 5-Me-C, G,5- Me-U

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moeBs B = nucleobase A, 5-Me-C, G,5- Me-U

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4alkdBs B = nucleobase A, C, G, U

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araBs B = nucleobase A, C, G, U

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meFBs B = nucleobase A, C, G, U

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cf3Bs B = nucleobase A, C, G, U

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25Bs B = nucleobase A, C, G, U

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rxyloBs B = nucleobase A, C, G, U

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amiBs B = nucleobase A, C, G, U

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ome4SBs B = nucleobase A, C, G, U

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!TABLE 3a

Position 2-23 nucleotides or derivatives thereof

Nucleotide

fara8azA

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fara7dzA

r2AP

d2aminoA

r7dz8azA

4S8azG

d5propU

d5propC

r7dzA

d7dzA

fara8azAs

fara7dzAs

r2APs

d2aminoAs

r7dz8azAs

4S8azGs

d5propUs

d5propCs

r7dzAs

d7dzAs

An embodiment of this invention is realized when nucleotide position 2 starting from the 5′ end is selected from the group consisting of rC, rCs, fluC, fluCs, OMeC, and OMeCs. A subembodiment of this aspect of the invention is realized when nucleotide position 2 starting from the 5′ end is rC or rCs. Another subembodiment of this aspect of the invention is realized when nucleotide position 2 starting from the 5′ end is fluC or fluCs. Another subembodiment of this aspect of the invention is realized when nucleotide position 2 starting from the 5′ end is OMeC or OMeCs.

TABLE 4

Nomenclature for nucleotide position 24 starting from 5′ exemplified in Table 5

Nucleotide

Nucleotide

position 24 -
Structure
Position 24
Structure

rCSup

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rC;pSup

fluC Sup

fluC;pSup

omeCSup

omeC;pSup

An embodiment of this invention is realized when R₃of the position 24 nucleotide is hydrogen, K is hydrogen or P(O)(OH)₂, X is O, and one of R₁and R₂is hydrogen while the other is selected from F, OH, and OMe. An embodiment of this invention is realized when the position 24 nucleotide is selected from Table 4. A subembodiment of this aspect of the invention is realized when nucleotide position 24 is rCSup or rC;pSup. Another subembodiment of this aspect of the invention is realized when nucleotide position 24 starting from the 5′ end is fluCSup or fluC;pSup. Another subembodiment of this aspect of the invention is realized when nucleotide position 24 starting from the 5′ end is omeCSup or omeC;pSup.

An embodiment of this invention is realized when position 23 nucleotide starting from the 5′ end of Formula Ia and Formula I is selected from rG, rGs, fluG, fluGs, omeG, and omeGs. A subembodiment of this aspect of the invention is realized when nucleotide position 23 is rG or rGs. Another subembodiment of this aspect of the invention is realized when nucleotide position 23 starting is fluG or fluGs. Another subembodiment of this aspect of the invention is realized when nucleotide position 23 is omeG or omeGs.

Another embodiment of this invention is realized when starting from the 5′ end of Formula Ia and Formula I, position 1 nucleotide is rG or rGs, and position 24 nucleotide is selected from the group consisting of rCSup, rC;pSup, fluCSup, fluC;pSup, omeC;pSup, and omeCSup.

Another embodiment of this invention is realized when starting from the 5′ end of Formula Ia and Formula I, position 1 nucleotide is rG or rGs, position 2 nucleotide is selected from the group consisting of rC, rCs, fluC, fluCs, omeCs, and omeC, position 23 is selected from the group consisting of rG, rGs, fluG, fluGs, omeG, and omeGs, and position 24 nucleotide is selected from the group consisting of rCSup, rC;pSup, fluCSup, fluC;pSup, omeC;pSup, and omeCSup.

General Synthetic Scheme

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Oligonucleotides were prepared by chemical synthesis, as presented in Scheme 1. The oligonucleotide target sequence was synthesized on solid support, such as on controlled pore glass. After cleavage of the protected oligonucleotide from the solid support and deprotection of all protecting groups, each strand was purified chromatographically with a reversed phase (C18) or anion exchange (SAX) resin. After purification, each oligonucleotide product was lyophilized to dryness.

The synthesis of each oligonucleotide was accomplished on a solid support, such as controlled pore glass, using a commercially available automated oligosynthesizers. The solid support was obtained pre-loaded with the first (3′) nucleoside unit of the desired sequence and placed in an appropriate column position for the oligosynthesizer. The first nucleoside was linked to the solid support via a succinate linkage and contained a suitable acid sensitive protecting group (e.g., trityl, dimethoxytrityl) on the 5′-hydroxyl group. The solid-phase oligosynthesis employed synthetic procedures that are generally known in the art. Elongation of the desired oligomeric sequence went through a cycle of four steps: 1) Acidic deprotection of the 5′-trityl protecting group; 2) Coupling of the next nucleotide unit as the 5′-trityl (or dimethoxytrityl) protected phosphoramidite in the presence of an activating agent, such as S-ethyl-tetrazole; 3) Oxidation of the P(III) phosphite triester to the P(V) phosphate triester by an oxidizing agent, such as iodine; and 4) Capping any remaining unreacted alcohol groups through esterification with an acylating agent, such as acetic anhydride. The phosphoramidites used were either derived from naturally occurring nucleotide units or from chemically modified versions of these nucleotides. Typically, all phosphoramidites were prepared in solutions of acetonitrile (or other suitable solvents or solvent mixtures, such as acetonitrile with some percentage of dimethyl formamide or some other appropriate organic solvent). The activator for phosphoramidite coupling was typically dissolved in acetonitrile. An oxidizing agent, such as iodine, was dissolved in a suitable solvent mixture, such as acetonitrile, pyridine and water. Acidic detritylation reagents, such as dichloroacetic acid or trichloroacetic acid, were dissolved in appropriate solvents, such as toluene or dichloromethane. Acylating capping reagents were, for example, a mixture of acetic anhydride, 2,6-lutidine and N-methyl-imidazole in acetonitrile. In place of the oxidation of the phosphite triester to the phosphate triester, the P(III) intermediate may be converted to the phosphorothioate analog with a sulfurizing reagent, such as phenyl-acetyl-disulfide, in a suitable solvent, such as a mixture of acetonitrile and pyridine. In between each step of the oligonucleotide elongation, acetonitrile (or another suitable solvent) was used to remove excess reagents and wash the solid support.

Oligonucleotide synthesis cycles were continued until the last (5′) nucleotide unit was installed onto the extended oligomer. After the final cycle, the 5′-trityl protecting group may or may not be removed from the oligonucleotide while it remains on the solid support. In some instances, the 5′-terminal trityl group was first removed by treatment with an acidic solution. After this deprotection, the solid support was treated with an appropriate base, such as aqueous methyl amine or ammonia:methyl amine (1:1) (at either room temperature or with mild heating) in order to cleave the oligonucleotide from the support, remove the cyanoethyl protecting groups on the phosphate linkages and deprotect the acyl protecting groups on the nucleotide bases. Purification of these oligonucleotides would then be accomplished by SAX or reversed phase (C18 or C8) chromatography. Typically, the oligonucleotide was eluted from the SAX resin with a gradient of an inorganic salt, such as sodium chloride or sodium perchlorate. Salt was removed from the purified samples by dialysis or tangential flow filtration. The desalted material was then lyophilized or annealed. Alternatively, the purified oligonucleotides were annealed prior to removal of salt and then dialyzed.

In order to synthesize certain 5′-modified oligonucleotides, such as those containing 5′-ppp and 5′-pp, additional synthetic steps were performed on the oligonucleotide prior to cleavage from the solid support. After removal of the 5′-protecting group of the terminal nucleotide, a suitable phosphoramidite or comparable reagent was used to generate an activated monophosphate or phosphite intermediate. The intermediate was oxidized, followed by coupling with pyrophosphate or phosphate, or analogs thereof, to provide the desired 5′-ppp or 5′-pp oligonucleotide after cleavage and deprotection of the oligonucleotide under appropriate conditions. Alternatively, the intermediate was coupled to pyrophosphate or phosphate, or analogs thereof, followed by the oxidation step.

In other instances, the 5′-trityl protecting group was left on the oligonucleotide during basic cleavage from the solid support and deprotection of the oligonucleotide protecting groups. In this case, the oligonucleotide was purified with a reversed phase resin, such as C18. The presence of the trityl group allowed for the desired full length oligonucleotide to be retained on the resin, while undesired truncated products that were acylated during the capping reaction were removed from the resin. These undesired oligonucleotides were removed from the resin with a lower percentage of an organic solvent, such as acetonitrile. After this removal of failure products, the trityl group was removed by acidic treatment (with an acid such as aqueous trifluoroacetic acid) while still on the C18 resin. After salt exchange and extensive washing of the resin with water, the desired deprotected product was eluted with a higher percentage of organic solvent in water. These purified oligonucleotides were then either lyophilized or annealed as required.

If the desired oligonucleotide contained any ribose (2′-hydroxyl) nucleotides, a modified procedure was required to remove the silyl 2′-hydroxyl protecting groups. After cleaving the oligonucleotide from the solid support with aqueous base, the column was further washed with an appropriate solvent, such as DMSO, to remove any material remaining on the column. After basic deprotection of the oligonucleotide, the reaction mixture then was treated with an appropriate fluoride reagent, such as triethylamine-hydrogen fluoride or TBAF, to cleave all of the silyl ethers and expose the desired alcohols. After silyl deprotection was complete, an appropriate buffer was added to each sample in order to neutralize the solution prior to purification (either by SAX or C18). In certain cases, a 5′-vinyl phosphonate-3′-phosphoramidite was coupled to the 5′-terminus of an oligonucleotide. The oxidation reagent for this incorporation may be changed to t-butyl hydroperoxide rather than iodine. When an oligonucleotide contained a protected phosphonate moiety, the methyl or ethyl phosphate esters were deprotected with an appropriate reagent, such as iodotrimethylsilane or bromotrimethylsilane in pyridine/acetonitrile, while the oligonucleotide was still on the solid support. After washing the support with acetonitrile, the oligonucleotide was treated with (3-mercaptoethanol in triethylamine/acetonitrile. After further washing with acetonitrile, the oligonucleotide product was cleaved from the solid support and fully deprotected with aqueous methylamine. The 5′-phosphonate oligonucleotides were purified by either SAX or C18 chromatography.

Each purified oligonucleotide was analyzed for purity by appropriate methods, including reversed phase HPLC, SAX HPLC, and capillary gel electrophoresis. The identity of the oligonucleotide was confirmed by mass spectrometry, using an ionization technique such as ESI or MALDI. The yields of each oligonucleotide were assessed by UV (260 nm) with a theoretically derived extinction coefficient. The appropriate amounts of each strand were approximated by UV (260 nm) measurements and theoretical EC260 extinction coefficients. The final oligonucleotide material was lyophilized or dissolved in an appropriate aqueous buffer prior to biochemical or biological evaluation.

Example 1

5′-HO;rG;rC;rU;rC;rA;rU;rU;rU;rA;rC;rC;rA;rG;rC;rG;rU;rA;rA;rA;rU;rG;rA;rG;rC-3′(SEQ ID NO. 838)

Oligosynthesis was conducted on a MerMade12 oligosynthesizer at 10 μmol scale. Suitably protected ribonucleoside 3′-phosphoramidites (5′-ODMT, 2′-OTBS, N-acetyl-cytidine, N-benzoyl-adenosine, N-isobutyryl-guanosine) were dissolved in acetonitrile (0.1 M). Ethylthio-tetrazole (ETT) was used as activator (0.25 M in acetonitrile). A solution of iodine (0.025 M) in pyridine/acetonitrile/water (45/45/10) was used as oxidizer. Detritylation was accomplished with a solution of 3% trichloroacetic acid (TCA) in methylene chloride. The CPG solid support was preloaded with the 3′-terminal nucleoside, 5′-ODMT-2′-OTBS-N-acetyl-cytidine. The DMT protecting group was removed with three cycles of 3% TCA, followed by 3×2 mL washes of the support with acetontrile. Equal volumes (0.5 mL each) of the required amidite solution (corresponding to the desired oligonucleotide sequence from the 3′-terminus) and activator was next added to the CPG. Nucleotide coupling was accomplished with three cycles of amidite/activator for 7.5 minutes each. After completion of the amidite coupling and washing of the CPG with acetonitrile (3×2 mL), the P(III) intermediate was oxidized to the P(V) phosphate ester with iodine (4×1 mL, 1 min each) and washed with acetonitrile (3×2 mL). Unreacted oligonucleotide 5′-hydroxyls were capped with an equal volume mixture (0.5 mL each) of 20% acetic anhydride/30% lutidine in acetonitrile and N-methyl-imidazole (20% in acetonitrile). The solid support was then washed with acetonitrile (4×2 mL), before the next coupling cycle was initiated with detritylation of the newly installed nucleotide unit.

After completion of the synthesis of the oligonucleotide sequence, the CPG support was washed with acetonitrile (4×2 mL). The protected oligonucleotide was removed from the solid support with 40% aqueous methylamine (3 mL, then 2 mL) and DMSO (2×3 mL). The combined solutions were heated at 35° C. for 45 minutes. The oligonucleotide was then treated with 3 mL of HF-triethylamine and heated to 40° C. for 60 minutes prior to the addition of 50 mM aqueous NaOAc (15 mL). The reaction mixture was directly loaded onto a C18 cartridge (5 g) that was prewashed with acetonitrile, water and 50 mM aqueous triethylammonium acetate (TEAA). After washing with water and 16% acetonitrile in 50 mM TEAA/50 mM NaCl (to remove shortmer synthesis failures), the DMT group was deprotected on-column with 1% TFA. The column was washed with water, 1 M NaCl and additional water prior to elution of the desired oligonucleotide with 30% aqueous acetonitrile.

The desired oligonucleotide was further purified by preparative mass-directed HPLC with a Waters X-Bridge Phenyl column (19×150 mm, ambient temperature, 25 mL/min, buffer A: 200 mM aqueous TEAA, pH 7, buffer B: 200 mM TEAA, 20% aq. acetonitrile, pH 7) eluting with a gradient of 3% to 11% B over 9 minutes and 11% to 25% B over 3 minutes. The counterion of the oligonucleotide was exchanged to sodium via spin filtration (MWCO 3000) and the isolated product was lyophilized to a white solid.

LC-MS: (m/z)⁻ (z=4) 1908.1 (exp); 1908.1 (theor); (z=5) 1526.1 (exp); 1526.3 (theor)

Examples 2-336, 342-352, 360-404, 436-442, 475-477, 514-593, 595-605, 607, 609-613, 615, 617-633, 635-645, 647, 649-653, 655, and 657-838 noted as SEQ ID No. 1 as shown in Table 5 below were prepared according to the procedures analogous to those outlined in Example 1. Examples 337-341, 353-359, 405-435, 443-474, 478-513, 594, 606, 608, 614, 616, 634, 646, 648, 654, and 656 with modified nucleobases in Table 5 below were prepared using procedures known in the art in combination with the procedures analogous to those outlined in Example 1.

TABLE 5

SEQ

Ex
Sequence
MW
ID No.

1
5′-HO;rG;rC;rU;rC;rA;rU;rU;rU;rA;rC;rC;rA;rG;rC;rG;rU;
7636.72
1

rA;rA;rA;rU;rG;rA;rG;rCSup-3′

2
5′-HO;rG;fluC;rU;fluC;fluA;fluU;rU;fluU;rA;rC;fluC;rA;
7672.62
124

fluG;rC;fluG;fluU;fluA;rA;rA;rU;omeG;rA;rG;fluCSup-3′

3
5′-HO;rG;fluC;rU;fluC;rA;fluU;omeU;rU;fluA;rC;rC;rA;
7694.68
125

fluG;rC;rG;rU;rA;omeA;fluA;fluU;rG;rA;fluG;omeCSup-3′

4
5′-HO;rG;rC;fluU;rC;fluA;rU;rU;omeU;fluA;fluC;rC;rA;
7706.71
126

fluG;fluC;fluG;rU;rA;omeA;omeA;rU;rG;rA;rG;omeCSup-3′

5
5′-HO;rG;rC;omeU;rC;rA;fluU;rU;omeU;fluA;rC;fluC;omeA;
7710.63
127

rG;rC;rG;fluU;rA;fluA;omeA;rU;fluG;fluA;fluG;fluCSup-3′

6
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7716.81
128

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

7
5′-HO;rG;omeC;rU;fluC;rA;rU;fluU;omeU;fluA;omeC;fluC;fluA;
7720.73
129

rG;fluC;omeG;fluU;rA;fluA;fluA;fluU;fluG;fluA;fluG;fluCSup-3′

8
5′-HO;rG;rC;fluU;rC;fluA;omeU;fluU;rU;rA;fluC;omeC;omeA;
7720.73
130

rG;fluC;omeG;omeU;rA;rA;rA;rU;fluG;rA;fluG;rCSup-3′

9
5′-HO;rG;rC;rU;rC;fluA;omeU;omeU;fluU;rA;fluC;rC;fluA;
7722.70
131

rG;fluC;omeG;rU;rA;rA;fluA;fluU;omeG;fluA;rG;omeCSup-3′

10
5′-HO;rG;rC;fluU;rC;fluA;fluU;fluU;omeU;fluA;omeC;rC;rA;
7724.64
132

omeG;omeC;fluG;rU;rA;fluA;omeA;fluU;fluG;rA;rG;rCSup-3′

11
5′-HO;rG;rC;omeU;omeC;fluA;rU;rU;rU;fluA;fluC;fluC;rA;
7724.71
133

omeG;fluC;fluG;rU;omeA;rA;fluA;rU;fluG;fluA;omeG;rCSup-3′

12
5′-HO;rG;rC;omeU;fluC;rA;rU;rU;rU;fluA;fluC;rC;fluA;fluG;
7724.68
134

rC;fluG;rU;rA;fluA;fluA;omeU;omeG;omeA;omeG;fluCSup-3′

13
5′-HO;rG;omeC;fluU;rC;omeA;fluU;omeU;omeU;fluA;omeC;
7726.92
135

fluC;rA;fluG;fluC;rG;rU;fluA;rA;fluA;fluU;rG;fluA;rG;rCSup-3′

14
5′-HO;rG;rC;fluU;fluC;fluA;omeU;fluU;rU;fluA;omeC;fluC;fluA;
7728.71
136

rG;fluC;rG;omeU;rA;rA;fluA;fluU;rG;omeA;omeG;fluCSup-3′

15
5′-HO;rG;fluC;rU;omeC;rA;omeU;rU;omeU;omeA;fluC;rC;fluA;
7728.67
137

fluG;rC;fluG;omeU;fluA;rA;fluA;rU;fluG;fluA;fluG;fluCSup-3′

16
5′-HO;rG;fluC;rU;rC;fluA;rU;fluU;fluU;fluA;rC;rC;omeA;fluG;
7730.58
138

fluC;omeG;fluU;omeA;omeA;fluA;omeU;fluG;fluA;fluG;rCSup-3′

17
5′-HO;rG;omeC;fluU;omeC;rA;rU;rU;rU;rA;fluC;rC;rA;rG;
7730.78
139

rC;omeG;fluU;rA;fluA;fluA;omeU;omeG;omeA;rG;rCSup-3′

18
5′-HO;rG;rC;omeU;rC;omeA;rU;fluU;fluU;rA;rC;omeC;omeA;
7730.75
140

rG;fluC;omeG;rU;rA;rA;rA;fluU;fluG;rA;omeG;rCSup-3′

19
5′-HO;rG;rC;rU;rC;omeA;fluU;omeU;rU;rA;rC;fluC;omeA;
7732.73
141

omeG;rC;rG;fluU;rA;fluA;omeA;rU;rG;fluA;omeG;fluCSup-3′

20
5′-HO;rG;rC;rU;fluC;fluA;fluU;omeU;rU;rA;fluC;omeC;rA;rG;
7734.77
142

omeC;rG;omeU;fluA;omeA;rA;omeU;fluG;rA;fluG;rCSup-3′

21
5′-HO;rG;omeC;fluU;rC;fluA;fluU;fluU;rU;rA;fluC;rC;rA;rG;
7736.75
143

fluC;rG;omeU;omeA;omeA;fluA;rU;omeG;rA;omeG;fluCSup-3′

22
5′-HO;rG;rC;rU;rC;fluA;rU;fluU;rU;fluA;omeC;fluC;omeA;rG;
7736.71
144

fluC;omeG;fluU;fluA;rA;omeA;omeU;omeG;rA;fluG;rCSup-3′

23
5′-HO;rG;omeC;fluU;rC;omeA;omeU;rU;fluU;omeA;rC;fluC;rA;
7736.68
145

fluG;rC;fluG;omeU;fluA;fluA;rA;omeU;fluG;rA;rG;rCSup-3′

24
5′-HO;rG;omeC;rU;rC;rA;rU;fluU;rU;omeA;rC;omeC;omeA;fluG;
7736.71
146

fluC;rG;fluU;rA;omeA;fluA;fluU;fluG;fluA;omeG;rCSup-3′

25
5′-HO;rG;omeC;omeU;rC;fluA;fluU;fluU;omeU;rA;omeC;omeC;
7736.75
147

rA;rG;rC;fluG;fluU;rA;fluA;fluA;omeU;rG;rA;rG;fluCSup-3′

26
5′-HO;rG;rC;fluU;omeC;omeA;fluU;rU;fluU;rA;omeC;fluC;rA;
7736.75
148

omeG;rC;rG;rU;rA;rA;fluA;fluU;omeG;omeA;fluG;fluCSup-3′

27
5′-HO;rG;omeC;fluU;rC;rA;omeU;fluU;fluU;fluA;omeC;rC;rA;
7738.73
149

rG;fluC;omeG;omeU;fluA;fluA;fluA;rU;fluG;rA;rG;omeCSup-3′

28
5′-HO;rG;omeC;omeU;fluC;fluA;rU;fluU;rU;rA;rC;rC;fluA;omeG;
7738.69
150

rC;rG;fluU;omeA;fluA;fluA;fluU;omeG;fluA;rG;omeCSup-3′

29
5′-HO;rG;fluC;fluU;rC;fluA;omeU;rU;rU;fluA;rC;rC;fluA;rG;
7738.69
151

omeC;omeG;omeU;fluA;fluA;omeA;rU;fluG;fluA;rG;omeCSup-3′

30
5′-HO;rG;fluC;omeU;rC;omeA;fluU;omeU;fluU;fluA;omeC;omeC;
7740.67
152

rA;fluG;rC;fluG;rU;omeA;fluA;fluA;rU;fluG;rA;fluG;rCSup-3′

31
5′-HO;rG;fluC;fluU;omeC;rA;fluU;fluU;rU;rA;fluC;fluC;omeA;
7742.75
153

omeG;omeC;fluG;fluU;rA;rA;fluA;omeU;fluG;fluA;rG;omeCSup-3′

32
5′-HO;rG;rC;rU;omeC;omeA;fluU;fluU;fluU;omeA;omeC;fluC;
7746.77
154

fluA;rG;rC;rG;omeU;rA;omeA;rA;omeU;rG;fluA;rG;rCSup-3′

33
5′-HO;rG;rC;omeU;omeC;rA;omeU;fluU;rU;omeA;rC;fluC;rA;
7746.81
155

omeG;fluC;omeG;rU;rA;fluA;rA;rU;fluG;rA;omeG;fluCSup-3′

34
5′-HO;rG;omeC;rU;omeC;omeA;omeU;omeU;fluU;fluA;fluC;rC;
7748.82
156

fluA;rG;fluC;rG;rU;rA;rA;omeA;rU;rG;fluA;omeG;fluCSup-3′

35
5′-HO;rG;fluC;rU;omeC;fluA;omeU;omeU;fluU;rA;fluC;fluC;rA;
7750.76
157

rG;rC;omeG;fluU;omeA;rA;fluA;fluU;rG;omeA;omeG;rCSup-3′

36
5′-HO;rG;fluC;fluU;omeC;rA;rU;fluU;omeU;fluA;rC;rC;rA;fluG;
7750.76
158

fluC;omeG;rU;fluA;omeA;rA;rU;omeG;omeA;omeG;fluCSup-3′

37
5′-HO;rG;rC;fluU;omeC;omeA;rU;fluU;fluU;rA;omeC;fluC;fluA;
7752.74
159

fluG;rC;omeG;fluU;omeA;rA;fluA;omeU;rG;rA;fluG;omeCSup-3′

38
5′-HO;rG;omeC;omeU;fluC;omeA;rU;fluU;rU;fluA;omeC;rC;rA;rG;
7752.77
160

omeC;fluG;fluU;fluA;rA;fluA;rU;fluG;fluA;omeG;omeCSup-3′

39
5′-HO;rG;rC;rU;rC;rA;rU;omeU;omeU;rA;fluC;fluC;fluA;omeG;
7752.74
161

fluC;omeG;fluU;fluA;omeA;fluA;fluU;omeG;rA;omeG;fluCSup-3′

40
5′-HO;rG;rC;fluU;rC;rA;rU;fluU;fluU;omeA;omeC;rC;omeA;fluG;
7752.67
162

rC;fluG;omeU;fluA;omeA;fluA;fluU;rG;omeA;fluG;omeCSup-3′

41
5′-HO;rG;fluC;omeU;omeC;omeA;rU;fluU;omeU;rA;fluC;rC;rA;
7754.75
163

omeG;fluC;rG;omeU;rA;fluA;omeA;fluU;fluG;fluA;fluG;fluCSup-3′

42
5′-HO;rG;rC;omeU;omeC;fluA;fluU;fluU;rU;rA;fluC;fluC;omeA;
7756.73
164

fluG;fluC;omeG;rU;fluA;fluA;rA;fluU;omeG;fluA;omeG;omeCSup-3′

43
5′-HO;rG;omeC;rU;rC;omeA;fluU;fluU;fluU;rA;omeC;fluC;omeA;
7756.73
165

rG;omeC;fluG;rU;omeA;fluA;fluA;fluU;fluG;omeA;fluG;fluCSup-3′

44
5′-HO;rG;rC;fluU;omeC;rA;omeU;rU;omeU;omeA;rC;omeC;
7756.83
166

rA;rG;rC;fluG;rU;omeA;omeA;omeA;rU;rG;rA;fluG;fluCSup-3′

45
5′-HO;rG;rC;rU;fluC;rA;rU;rU;omeU;omeA;omeC;omeC;rA;
7756.87
167

fluG;fluC;omeG;omeU;rA;rA;fluA;omeU;rG;rA;omeG;rCSup-3′

46
5′-HO;rG;omeC;fluU;fluC;omeA;fluU;rU;fluU;omeA;fluC;fluC;
7758.75
168

omeA;fluG;fluC;rG;omeU;omeA;rA;fluA;omeU;fluG;rA;fluG;fluCSup-3′

47
5′-HO;rG;rC;omeU;fluC;rA;rU;rU;rU;rA;rC;omeC;fluA;fluG;fluC;
7760.82
169

rG;omeU;fluA;omeA;omeA;omeU;fluG;omeA;rG;omeCSup-3′

48
5′-HO;rG;fluC;rU;rC;rA;omeU;omeU;fluU;fluA;rC;fluC;omeA;
7760.79
170

omeG;rC;fluG;omeU;rA;rA;omeA;rU;omeG;rA;fluG;omeCSup-3′

49
5′-HO;rG;rC;rU;fluC;omeA;omeU;fluU;rU;fluA;rC;rC;fluA;omeG;
7762.76
171

omeC;rG;rU;fluA;rA;omeA;omeU;fluG;omeA;omeG;fluCSup-3′

50
5′-HO;rG;fluC;rU;omeC;omeA;fluU;omeU;rU;omeA;rC;rC;rA;fluG;
7762.76
172

fluC;rG;omeU;fluA;fluA;omeA;omeU;omeG;fluA;rG;rCSup-3′

51
5′-HO;rG;omeC;rU;omeC;omeA;omeU;rU;omeU;fluA;omeC;fluC;
7762.87
173

rA;fluG;fluC;rG;fluU;fluA;rA;omeA;fluU;rG;rA;rG;omeCSup-3′

52
5′-HO;rG;fluC;fluU;omeC;omeA;rU;omeU;rU;rA;rC;fluC;fluA;rG;
7764.81
174

omeC;fluG;omeU;rA;omeA;fluA;rU;omeG;fluA;omeG;fluCSup-3′

53
5′-HO;rG;fluC;omeU;fluC;fluA;rU;rU;omeU;rA;fluC;fluC;omeA;rG;
7764.85
175

fluC;omeG;rU;omeA;rA;omeA;omeU;rG;fluA;omeG;fluCSup-3′

54
5′-HO;rG;fluC;fluU;fluC;omeA;omeU;fluU;rU;rA;fluC;omeC;rA;
7766.79
176

fluG;rC;omeG;omeU;rA;fluA;omeA;fluU;rG;fluA;omeG;omeCSup-3′

55
5′-HO;rG;fluC;fluU;fluC;rA;fluU;fluU;rU;omeA;fluC;fluC;omeA;
7768.80
177

omeG;omeC;rG;fluU;omeA;fluA;rA;rU;omeG;omeA;fluG;omeCSup-3′

56
5′-HO;rG;omeC;rU;fluC;omeA;fluU;fluU;omeU;fluA;rC;omeC;fluA;
7768.77
178

rG;fluC;fluG;omeU;fluA;rA;omeA;omeU;rG;omeA;fluG;fluCSup-3′

57
5′-HO;rG;omeC;fluU;rC;fluA;omeU;rU;fluU;fluA;rC;omeC;fluA;
7768.73
179

omeG;fluC;fluG;rU;omeA;fluA;fluA;fluU;omeG;rA;omeG;omeCSup-3′

58
5′-HO;rG;rC;fluU;fluC;omeA;fluU;omeU;omeU;omeA;fluC;rC;omeA;
7770.71
180

fluG;omeC;omeG;fluU;fluA;fluA;fluA;fluU;rG;omeA;rG;fluCSup-3′

59
5′-HO;rG;omeC;fluU;rC;fluA;fluU;omeU;omeU;omeA;omeC;fluC;
7770.71
181

fluA;fluG;rC;omeG;fluU;fluA;fluA;rA;rU;omeG;fluA;omeG;fluCSup-3′

60
5′-HO;rG;fluC;omeU;omeC;fluA;omeU;fluU;fluU;fluA;fluC;omeC;fluA;
7772.76
182

fluG;omeC;omeG;fluU;rA;omeA;rA;omeU;fluG;fluA;rG;fluCSup-3′

61
5′-HO;rG;fluC;omeU;rC;fluA;omeU;rU;omeU;rA;rC;omeC;fluA;rG;
7776.81
183

omeC;rG;fluU;omeA;omeA;rA;fluU;fluG;omeA;fluG;omeCSup-3′

62
5′-HO;rG;fluC;rU;rC;fluA;fluU;omeU;omeU;fluA;fluC;omeC;omeA;
7776.81
184

omeG;rC;omeG;omeU;rA;rA;fluA;rU;fluG;rA;omeG;omeCSup-3′

63
5′-HO;rG;omeC;omeU;omeC;rA;rU;fluU;fluU;rA;rC;omeC;omeA;
7776.85
185

rG;omeC;rG;omeU;fluA;fluA;fluA;fluU;omeG;rA;omeG;fluCSup-3′

64
5′-HO;rG;omeC;fluU;omeC;rA;omeU;rU;omeU;fluA;omeC;fluC;fluA;
7776.81
186

fluG;rC;rG;omeU;rA;rA;omeA;fluU;omeG;fluA;omeG;rCSup-3′

65
5′-HO;rG;rC;rU;fluC;omeA;fluU;rU;rU;fluA;omeC;omeC;rA;omeG;
7776.85
187

omeC;omeG;omeU;rA;fluA;fluA;omeU;fluG;omeA;rG;fluCSup-3′

66
5′-HO;rG;fluC;omeU;fluC;rA;omeU;rU;fluU;rA;omeC;omeC;omeA;
7778.82
188

rG;rC;omeG;fluU;fluA;fluA;fluA;omeU;fluG;omeA;rG;omeCSup-3′

67
5′-HO;rG;fluC;omeU;fluC;fluA;fluU;fluU;rU;rA;rC;omeC;omeA;fluG;
7778.82
189

fluC;omeG;rU;rA;omeA;omeA;omeU;omeG;fluA;rG;omeCSup-3′

68
5′-HO;rG;omeC;fluU;fluC;fluA;omeU;omeU;rU;omeA;rC;omeC;omeA;
7778.79
190

omeG;fluC;omeG;rU;fluA;fluA;fluA;omeU;rG;fluA;rG;rCSup-3′

69
5′-HO;rG;rC;omeU;fluC;omeA;omeU;rU;rU;fluA;fluC;fluC;omeA;
7778.79
191

omeG;rC;rG;fluU;fluA;omeA;fluA;omeU;rG;omeA;omeG;fluCSup-3′

70
5′-HO;rG;omeC;rU;fluC;rA;omeU;fluU;rU;fluA;fluC;omeC;rA;omeG;
7780.80
192

rC;omeG;omeU;fluA;omeA;omeA;fluU;fluG;omeA;fluG;fluCSup-3′

71
5′-HO;rG;rC;omeU;fluC;rA;omeU;omeU;fluU;fluA;fluC;omeC;fluA;
7780.80
193

fluG;omeC;fluG;fluU;rA;fluA;omeA;rU;rG;omeA;omeG;omeCSup-3′

72
5′-HO;rG;omeC;omeU;fluC;rA;fluU;rU;rU;omeA;fluC;fluC;fluA;rG;
7780.80
194

omeC;fluG;omeU;omeA;omeA;fluA;omeU;fluG;fluA;omeG;rCSup-3′

73
5′-HO;rG;fluC;omeU;rC;omeA;omeU;rU;fluU;rA;omeC;omeC;omeA;
7782.78
195

fluG;fluC;fluG;rU;fluA;fluA;omeA;fluU;omeG;omeA;fluG;fluCSup-3′

74
5′-HO;rG;fluC;fluU;omeC;fluA;fluU;omeU;rU;fluA;omeC;rC;fluA;fluG;
7784.76
196

omeC;rG;fluU;omeA;omeA;fluA;omeU;fluG;omeA;omeG;fluCSup-3′

75
5′-HO;rG;omeC;fluU;fluC;fluA;omeU;fluU;fluU;omeA;fluC;omeC;fluA;
7784.75
197

omeG;rC;fluG;fluU;omeA;rA;omeA;rU;omeG;fluA;fluG;omeCSup-3′

76
5′-HO;rG;omeC;rU;omeC;rA;omeU;fluU;fluU;rA;rC;fluC;omeA;
7784.89
198

fluG;omeC;omeG;rU;omeA;rA;omeA;rU;omeG;omeA;rG;rCSup-3′

77
5′-HO;rG;rC;rU;omeC;omeA;fluU;rU;rU;omeA;omeC;omeC;fluA;
7786.90
199

rG;omeC;omeG;rU;fluA;rA;fluA;omeU;rG;omeA;fluG;omeCSup-3′

78
5′-HO;rG;fluC;omeU;fluC;fluA;omeU;omeU;omeU;omeA;omeC;
7786.91
200

omeC;fluA;omeG;fluC;rG;rU;rA;omeA;rA;rU;omeG;rA;rG;rCSup-3′

79
5′-HO;rG;rC;rU;fluC;omeA;omeU;fluU;omeU;fluA;omeC;rC;rA;fluG;
7786.83
201

omeC;omeG;rU;omeA;omeA;fluA;rU;omeG;rA;omeG;rCSup-3′

80
5′-HO;rG;fluC;omeU;fluC;omeA;fluU;omeU;omeU;rA;rC;rC;fluA;fluG;
7792.77
202

rC;omeG;omeU;omeA;fluA;fluA;rU;fluG;omeA;omeG;omeCSup-3′

81
5′-HO;rG;omeC;omeU;omeC;fluA;omeU;rU;rU;omeA;fluC;omeC;fluA;
7794.85
203

fluG;omeC;fluG;rU;fluA;omeA;omeA;fluU;fluG;rA;omeG;fluCSup-3′

82
5′-HO;rG;fluC;omeU;rC;omeA;omeU;fluU;rU;fluA;omeC;omeC;fluA;
7794.82
204

fluG;fluC;fluG;omeU;omeA;omeA;rA;fluU;omeG;omeA;rG;fluCSup-3′

83
5′-HO;rG;rC;fluU;omeC;fluA;omeU;rU;fluU;omeA;omeC;fluC;rA;omeG;
7794.81
205

fluC;fluG;omeU;fluA;fluA;rA;omeU;fluG;omeA;omeG;omeCSup-3′

84
5′-HO;rG;rC;omeU;rC;omeA;omeU;fluU;fluU;rA;rC;rC;omeA;omeG;
7798.80
206

rC;omeG;omeU;omeA;rA;fluA;omeU;omeG;fluA;rG;omeCSup-3′

85
5′-HO;rG;omeC;omeU;rC;rA;fluU;rU;rU;fluA;rC;fluC;omeA;omeG;
7798.90
207

omeC;omeG;omeU;omeA;rA;rA;fluU;rG;omeA;omeG;omeCSup-3′

86
5′-HO;rG;omeC;omeU;omeC;omeA;fluU;omeU;omeU;fluA;fluC;fluC;
7802.90
208

omeA;rG;omeC;omeG;fluU;fluA;omeA;omeA;rU;rG;rA;rG;rCSup-3′

87
5′-HO;rG;omeC;fluU;fluC;omeA;rU;omeU;fluU;fluA;omeC;fluC;omeA;
7802.93
209

fluG;omeC;rG;omeU;omeA;rA;omeA;rU;omeG;rA;rG;omeCSup-3′

88
5′-HO;rG;rC;omeU;omeC;rA;fluU;omeU;fluU;rA;omeC;omeC;rA;omeG;
7802.90
210

fluC;omeG;omeU;omeA;omeA;omeA;fluU;rG;fluA;rG;fluCSup-3′

89
5′-HO;rG;rC;fluU;omeC;omeA;rU;fluU;omeU;fluA;omeC;omeC;omeA;
7816.87
211

rG;rC;omeG;fluU;omeA;omeA;rA;omeU;fluG;fluA;omeG;omeCSup-3′

90
5′-HO;rG;omeC;fluU;omeC;omeA;omeU;rU;rU;fluA;fluC;omeComeA;
7820.86
212

omeG;fluC;omeG;omeU;omeA;fluA;fluA;fluU;omeG;omeA;fluG;rCSup-3′
Exact

7817.20

91
5′-HO;rG;rC;omeU;omeC;rA;fluU;omeU;omeU;fluA;omeC;rC;omeA;
7856.93
213

omeG;omeC;omeG;omeU;omeA;omeA;omeA;omeU;omeG;fluA;fluG;

fluCSup-3′

92
5′-HO;rG;fluC;fluU;fluC;fluA;fluU;fluU;fluU;fluA;fluC;fluC;fluA;fluG;
7682.40
214

fluC;fluG;fluU;fluA;fluA;fluA;fluU;fluG;fluA;fluG;fluCSup-3′

93
5′-HO;rG;rC;fluU;rC;rA;fluU;rU;omeU;rA;rC;omeC;fluA;
7692.66
215

omeG;fluC;rG;fluU;fluA;rA;rA;rU;rG;fluA;rG;rCSup-3′

94
5′-HO;rG;rC;fluU;rC;rA;fluU;fluU;rU;omeA;rC;fluC;fluA;
7696.58
216

rG;rC;fluG;rU;fluA;omeA;rA;fluU;omeG;fluA;rG;rCSup-3′

95
5′-HO;rG;rC;fluU;rC;rA;rU;omeU;rU;fluA;fluC;omeC;rA;rG;
7706.71
217

rC;rG;fluU;omeA;rA;rA;fluU;omeG;fluA;rG;fluCSup-3′

96
5′-HO;rG;fluC;omeU;fluC;rA;fluU;omeU;rU;rA;fluC;fluC;fluA;
7710.70
218

rG;rC;rG;rU;omeA;rA;fluA;fluU;omeG;rA;fluG;rCSup-3′

97
5′-HO;rG;fluC;rU;rC;fluA;rU;rU;rU;rA;fluC;fluC;omeA;omeG;
7712.72
219

fluC;fluG;omeU;fluA;omeA;rA;fluU;rG;fluA;rG;fluCSup-3′

98
5′-HO;rG;rC;fluU;fluC;omeA;fluU;fluU;fluU;fluA;rC;fluC;fluA;
7718.58
220

rG;fluC;fluG;fluU;rA;omeA;omeA;fluU;fluG;omeA;rG;rCSup-3′

99
5′-HO;rG;rC;omeU;rC;rA;omeU;fluU;fluU;fluA;fluC;rC;rA;fluG;
7720.69
221

rC;omeG;rU;omeA;rA;rA;omeU;rG;fluA;rG;fluCSup-3′

100
5′-HO;rG;fluC;fluU;fluC;rA;fluU;fluU;rU;rA;omeC;omeC;rA;
7724.79
222

omeG;fluC;fluG;rU;rA;rA;rA;rU;omeG;omeA;fluG;fluCSup-3′

101
5′-HO;rG;omeC;rU;fluC;fluA;fluU;fluU;fluU;fluA;fluC;rC;omeA;
7724.68
223

fluG;rC;rG;omeU;rA;fluA;rA;omeU;rG;omeA;rG;rCSup-3′

102
5′-HO;rG;fluC;rU;omeC;fluA;fluU;rU;rU;omeA;rC;rC;omeA;rG;
7728.67
224

fluC;fluG;fluU;rA;fluA;fluA;fluU;fluG;fluA;omeG;omeCSup-3′

103
5′-HO;rG;rC;rU;omeC;fluA;fluU;rU;fluU;rA;fluC;rC;rA;rG;
7732.79
225

omeC;rG;fluU;fluA;rA;rA;rU;omeG;omeA;omeG;omeCSup-3′

104
5′-HO;rG;omeC;omeU;rC;rA;rU;omeU;rU;fluA;rC;rC;fluA;
7732.73
226

omeG;fluC;rG;omeU;fluA;fluA;rA;omeU;rG;rA;fluG;rCSup-3′

105
5′-HO;rG;omeC;fluU;rC;rA;rU;fluU;rU;rA;fluC;rC;omeA;rG;
7734.77
227

omeC;fluG;rU;fluA;omeA;rA;omeU;rG;omeA;fluG;fluCSup-3′

106
5′-HO;rG;fluC;rU;rC;fluA;rU;rU;omeU;fluA;rC;fluC;rA;fluG;
7734.77
228

omeC;rG;rU;rA;fluA;rA;omeU;omeG;omeA;fluG;omeCSup-3′

107
5′-HO;rG;fluC;fluU;fluC;fluA;rU;rU;omeU;omeA;rC;rC;omeA;
7734.77
229

rG;omeC;fluG;rU;omeA;rA;rA;rU;rG;fluA;omeG;fluCSup-3′

108
5′-HO;rG;fluC;fluU;omeC;rA;rU;fluU;rU;omeA;omeC;fluC;rA;
7736.82
230

fluG;fluC;omeG;fluU;omeA;omeA;rA;rU;rG;rA;rG;fluCSup-3′

109
5′-HO;rG;rC;fluU;omeC;omeA;rU;fluU;omeU;rA;fluC;omeC;
7736.75
231

fluA;omeG;omeC;rG;rU;rA;fluA;fluA;rU;rG;fluA;fluG;rCSup-3′

110
5′-HO;rG;fluC;rU;rC;omeA;fluU;fluU;fluU;fluA;rC;omeC;rA;rG;
7736.75
232

omeC;rG;rU;omeA;fluA;rA;fluU;rG;fluA;omeG;omeCSup-3′

111
5′-HO;rG;rC;rU;fluC;rA;rU;rU;omeU;fluA;fluC;fluC;fluA;rG;
7736.79
233

omeC;fluG;rU;omeA;fluA;fluA;omeU;omeG;rA;rG;omeCSup-3′

112
5′-HO;rG;fluC;rU;fluC;omeA;rU;rU;omeU;fluA;fluC;omeC;
7736.75
234

omeA;rG;rC;fluG;fluU;fluA;omeA;fluA;rU;omeG;rA;rG;rCSup-3′

113
5′-HO;rG;rC;rU;rC;omeA;rU;fluU;fluU;omeA;fluC;fluC;rA;fluG;
7738.73
235

omeC;rG;fluU;omeA;omeA;omeA;fluU;fluG;rA;rG;fluCSup-3′

114
5′-HO;rG;rC;rU;fluC;fluA;rU;omeU;fluU;rA;omeC;rC;fluA;omeG;
7738.69
236

rC;fluG;rU;fluA;omeA;omeA;rU;fluG;fluA;fluG;omeCSup-3′

115
5′-HO;rG;omeC;fluU;fluC;fluA;fluU;rU;fluU;omeA;rC;fluC;fluA;
7738.73
237

rG;rC;omeG;rU;rA;omeA;omeA;omeU;fluG;rA;rG;fluCSup-3′

116
5′-HO;rG;omeC;rU;rC;rA;omeU;rU;fluU;rA;fluC;rC;fluA;fluG;
7738.69
238

fluC;omeG;omeU;omeA;omeA;fluA;rU;fluG;fluA;fluG;rCSup-3′

117
5′-HO;rG;omeC;fluU;fluC;rA;rU;omeU;fluU;omeA;omeC;fluC;fluA;
7740.78
239

fluG;fluC;rG;rU;omeA;fluA;rA;rU;fluG;fluA;rG;omeCSup-3′

118
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7744.80
240

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

119
5′-HO;rG;fluC;rU;fluC;rA;omeU;fluU;fluU;fluA;fluC;fluC;rA;omeG;
7746.71
241

omeC;fluG;omeU;fluA;omeA;fluA;fluU;rG;fluA;fluG;omeCSup-3′

120
5′-HO;rG;rC;fluU;omeC;rA;omeU;fluU;omeU;omeA;fluC;rC;
7746.77
242

omeA;fluG;rC;rG;omeU;rA;rA;rA;fluU;fluG;rA;rG;omeCSup-3′

121
5′-HO;rG;fluC;fluU;rC;omeA;rU;rU;rU;rA;omeC;omeC;fluA;fluG;
7750.76
243

omeC;omeG;omeU;fluA;rA;omeA;fluU;fluG;rA;fluG;rCSup-3′

122
5′-HO;rG;fluC;omeU;omeC;fluA;fluU;omeU;fluU;fluA;omeC;rC;fluA;
7752.74
244

rG;fluC;rG;omeU;omeA;fluA;rA;fluU;rG;rA;omeG;rCSup-3′

123
5′-HO;rG;rC;omeU;rC;fluA;omeU;fluU;omeU;omeA;rC;fluC;rA;fluG;
7752.67
245

omeC;fluG;fluU;fluA;rA;fluA;omeU;rG;fluA;omeG;rCSup-3′

124
5′-HO;rG;fluC;omeU;fluC;rA;rU;rU;fluU;omeA;omeC;rC;omeA;rG;
7752.74
246

fluC;fluG;omeU;fluA;fluA;omeA;fluU;rG;fluA;omeG;rCSup-3′

125
5′-HO;rG;fluC;omeU;omeC;omeA;rU;fluU;fluU;omeA;fluC;fluC;fluA;
7752.78
247

rG;omeC;rG;rU;fluA;fluA;omeA;fluU;rG;omeA;rG;rCSup-3′

126
5′-HO;rG;omeC;rU;fluC;rA;rU;omeU;rU;omeA;fluC;omeC;rA;fluG;
7754.79
248

fluC;omeG;fluU;fluA;fluA;rA;omeU;omeG;fluA;fluG;fluCSup-3′

127
5′-HO;rG;omeC;omeU;omeC;fluA;fluU;rU;omeU;fluA;fluC;rC;omeA;
7754.68
249

fluG;rC;fluG;fluU;rA;fluA;rA;fluU;omeG;omeA;fluG;rCSup-3′

128
5′-HO;rG;fluC;rU;omeC;rA;omeU;rU;rU;omeA;rC;omeC;fluA;
7754.86
250

omeG;rC;rG;rU;omeA;omeA;rA;fluU;rG;omeA;rG;rCSup-3′

129
5′-HO;rG;omeC;fluU;fluC;rA;fluU;omeU;omeU;rA;rC;fluC;fluA;fluG;
7756.73
251

omeC;omeG;fluU;fluA;rA;rA;fluU;fluG;fluA;omeG;omeCSup-3′

130
5′-HO;rG;omeC;omeU;omeC;omeA;rU;rU;omeU;fluA;rC;rC;
7756.83
252

fluA;omeG;rC;omeG;rU;fluA;rA;rA;rU;fluG;rA;rG;omeCSup-3′

131
5′-HO;rG;omeC;omeU;fluC;omeA;rU;rU;fluU;rA;rC;rC;omeA;
7756.83
253

omeG;rC;rG;fluU;rA;omeA;fluA;rU;rG;rA;omeG;omeCSup-3′

132
5′-HO;rG;rC;rU;rC;omeA;rU;rU;fluU;omeA;fluC;fluC;omeA;rG;
7758.85
254

fluC;rG;omeU;rA;omeA;fluA;rU;omeG;omeA;rG;omeCSup-3′

133
5′-HO;rG;rC;fluU;rC;rA;omeU;omeU;omeU;rA;omeC;rC;omeA;
7758.81
255

rG;omeC;rG;rU;fluA;omeA;fluA;omeU;rG;rA;fluG;fluCSup-3′

134
5′-HO;rG;fluC;omeU;fluC;omeA;omeU;rU;omeU;rA;rC;omeC;rA;
7760.86
256

rG;fluC;fluG;rU;omeA;rA;rA;omeU;fluG;fluA;rG;omeCSup-3′

135
5′-HO;rG;fluC;rU;rC;rA;omeU;omeU;omeU;rA;omeC;fluC;omeA;rG;
7760.79
257

rC;omeG;rU;fluA;fluA;fluA;rU;fluG;omeA;omeG;rCSup-3′

136
5′-HO;rG;fluC;fluU;rC;omeA;rU;omeU;fluU;omeA;fluC;rC;rA;omeG;
7760.75
258

rC;rG;rU;fluA;rA;omeA;omeU;fluG;omeA;omeG;rCSup-3′

137
5′-HO;rG;fluC;omeU;rC;rA;omeU;rU;fluU;fluA;fluC;fluC;rA;fluG;omeC;
7762.80
259

omeG;fluU;rA;rA;omeA;omeU;rG;omeA;omeG;rCSup-3′

138
5′-HO;rG;fluC;omeU;rC;omeA;rU;omeU;rU;omeA;omeC;fluC;fluA;
7762.84
260

omeG;fluC;fluG;fluU;rA;rA;rA;rU;rG;omeA;fluG;omeCSup-3′

139
5′-HO;rG;rC;fluU;omeC;fluA;rU;omeU;rU;fluA;rC;omeC;rA;fluG;rC;
7762.73
261

fluG;omeU;omeA;rA;omeA;fluU;omeG;fluA;omeG;rCSup-3′

140
5′-HO;rG;fluC;fluU;rC;rA;rU;rU;fluU;omeA;rC;omeC;fluA;omeG;omeC;
7763.80
262

omeG;fluU;rA;rA;omeA;omeU;fluG;rA;omeG;fluCSup-3′

141
5′-HO;rG;rC;fluU;omeC;rA;omeU;fluU;fluU;omeA;omeC;omeC;rA;
7764.77
263

rG;omeC;rG;fluU;fluA;fluA;omeA;rU;omeG;fluA;fluG;rCSup-3′

142
5′-HO;rG;rC;rU;rC;omeA;omeU;omeU;rU;omeA;fluC;rC;omeA;omeG;
7764.74
264

fluC;fluG;fluU;omeA;fluA;fluA;rU;fluG;omeA;rG;fluCSup-3′

143
5′-HO;rG;rC;omeU;fluC;rA;fluU;omeU;fluU;omeA;rC;rC;rA;omeG;
7766.68
265

fluC;fluG;fluU;fluA;omeA;fluA;fluU;omeG;omeA;omeG;rCSup-3′

144
5′-HO;rG;fluC;fluU;omeC;omeA;omeU;fluU;rU;rA;rC;rC;rA;omeG;
7766.75
266

fluC;fluG;fluU;omeA;omeA;omeA;omeU;rG;fluA;fluG;fluCSup-3′

145
5′-HO;rG;omeC;rU;omeC;fluA;omeU;omeU;fluU;rA;omeC;fluC;fluA;
7766.82
267

omeG;omeC;fluG;omeU;rA;fluA;rA;fluU;fluG;rA;rG;fluCSup-3′

146
5′-HO;rG;omeC;rU;omeC;fluA;fluU;fluU;fluU;omeA;fluC;omeC;omeA;
7768.80
268

fluG;fluC;rG;rU;fluA;rA;omeA;rU;fluG;omeA;fluG;omeCSup-3′

147
5′-HO;rG;omeC;omeU;fluC;omeA;fluU;rU;fluU;fluA;omeC;omeC;rA;
7770.78
269

omeG;omeC;omeG;fluU;fluA;fluA;rA;fluU;rG;fluA;fluG;fluCSup-3′

148
5′-HO;rG;fluC;fluU;fluC;omeA;omeU;omeU;omeU;fluA;fluC;omeC;
7770.78
270

fluA;rG;fluC;rG;fluU;omeA;fluA;rA;omeU;omeG;rA;fluG;fluCSup-3′

149
5′-HO;rG;omeC;fluU;rC;fluA;fluU;rU;rU;omeA;omeC;omeC;omeA;
7770.88
271

omeG;omeC;omeG;rU;omeA;rA;fluA;rU;rG;rA;rG;rCSup-3′

150
5′-HO;rG;rC;rU;fluC;rA;omeU;fluU;omeU;rA;rC;rC;omeA;omeG;fluC;
7770.81
272

omeG;fluU;omeA;rA;rA;omeU;omeG;rA;omeG;rCSup-3′

151
5′-HO;rG;omeC;omeU;omeC;rA;rU;omeU;omeU;rA;rC;fluC;omeA;
7772.82
273

omeG;rC;fluG;rU;fluA;fluA;omeA;rU;fluG;omeA;rG;rCSup-3′

152
5′-HO;rG;omeC;fluU;fluC;omeA;omeU;rU;omeU;rA;omeC;rC;omeA;
7772.82
274

fluG;rC;fluG;fluU;omeA;omeA;rA;rU;rG;omeA;rG;rCSup-3′

153
5′-HO;rG;fluC;rU;fluC;fluA;rU;rU;omeU;omeA;omeC;rC;omeA;rG;
7774.87
275

omeC;omeG;omeU;rA;rA;omeA;fluU;omeG;fluA;rG;fluCSup-3′

154
5′-HO;rG;rC;rU;omeC;omeA;rU;omeU;rU;fluA;rC;fluC;omeA;rG;rC;
7774.80
276

omeG;fluU;omeA;fluA;omeA;fluU;omeG;rA;fluG;omeCSup-3′

155
5′-HO;rG;fluC;rU;rC;rA;rU;fluU;omeU;rA;omeC;fluC;fluA;fluG;rC;
7776.81
277

rG;omeU;omeA;fluA;omeA;omeU;omeG;omeA;omeG;fluCSup-3′

156
5′-HO;rG;fluC;fluU;omeC;rA;rU;rU;fluU;fluA;omeC;omeC;omeA;omeG;
7776.81
278

rC;rG;fluU;omeA;omeA;rA;omeU;fluG;fluA;omeG;rCSup-3′

157
5′-HO;rG;fluC;omeU;fluC;rA;omeU;fluU;omeU;omeA;fluC;fluC;omeA;
7778.83
279

omeG;omeC;rG;rU;rA;fluA;omeA;fluU;fluG;rA;omeG;rCSup-3′

158
5′-HO;rG;rC;fluU;omeC;fluA;omeU;omeU;omeU;fluA;rC;omeC;omeA;
7778.75
280

fluG;omeC;fluG;fluU;rA;omeA;rA;fluU;rG;omeA;fluG;rCSup-3′

159
5′-HO;rG;rC;omeU;fluC;fluA;omeU;fluU;fluU;fluA;omeC;fluC;omeA;
7780.80
281

omeG;omeC;rG;omeU;fluA;fluA;rA;rU;omeG;rA;omeG;fluCSup-3′

160
5′-HO;rG;omeC;omeU;omeC;rA;fluU;fluU;omeU;fluA;fluC;rC;fluA;
7780.80
282

rG;fluC;fluG;fluU;omeA;rA;omeA;fluU;omeG;omeA;rG;omeCSup-3′

161
5′-HO;rG;omeC;rU;fluC;rA;rU;omeU;fluU;rA;omeC;omeC;rA;omeG;
7784.93
283

omeC;fluG;rU;rA;omeA;rA;omeU;omeG;fluA;omeG;rCSup-3′

162
5′-HO;rG;rC;rU;omeComeA;rU;omeU;omeU;omeA;fluC;omeC;omeA;
7788.85
284

fluG;rC;fluG;omeU;omeA;rA;rA;fluU;fluG;fluA;rG;omeCSup-3′

163
5′-HO;rG;omeC;rU;omeC;omeA;fluU;fluU;omeU;omeA;fluC;omeC;
7790.86
285

fluA;omeG;rC;fluG;omeU;fluA;rA;rA;omeU;rG;rA;omeG;fluCSup-3′

164
5′-HO;rG;omeC;fluU;omeC;fluA;omeU;omeU;fluU;fluA;rC;omeC;omeA;
7792.84
286

rG;fluC;rG;omeU;fluA;omeA;fluA;rU;omeG;omeA;rG;fluCSup-3′

165
5′-HO;rG;omeC;omeU;omeC;fluA;omeU;omeU;omeU;rA;omeC;fluC;
7794.81
287

rA;omeG;fluC;rG;fluU;omeA;omeA;fluA;fluU;fluG;fluA;fluG;rCSup-3′

166
5′-HO;rG;omeC;fluC;fluU;omeC;rA;fluU;omeU;fluU;fluA;fluC;omeC;
7794.85
288

omeA;omeG;fluC;omeG;rU;omeA;rA;fluA;fluU;omeG;omeA;rG;
found

omeCSup-3′

167
5′-HO;rG;omeC;omeU;fluC;fluA;omeU;fluU;omeU;omeA;fluC;fluC;
7796.79
289

omeA;fluG;rC;omeG;rU;fluA;omeA;omeA;fluU;rG;fluA;fluG;omeCSup-3′

168
5′-HO;rG;omeC;omeU;rC;fluA;omeU;omeU;rU;omeA;omeC;omeC;
7796.93
290

rA;rG;rC;rG;fluU;rA;rA;rA;omeU;omeG;omeA;fluG;omeCSup-3′

169
5′-HO;rG;omeC;omeU;fluC;fluA;rU;omeU;omeU;omeA;omeC;rC;
7800.88
291

rA;rG;rC;omeG;omeU;omeA;rA;omeA;fluU;fluG;omeA;rG;fluCSup-3′

170
5′-HO;rG;rC;omeU;fluC;rA;omeU;omeU;omeU;omeA;fluC;fluC;rA;
Calculated
292

rG;omeC;fluG;omeU;omeA;omeA;rA;fluU;rG;omeA;fluG;omeCSup-3′
7802.90

171
5′-HO;rG;omeC;omeU;rC;omeA;fluU;fluU;fluU;omeA;fluC;omeC;fluA;
7802.82
293

rG;rC;omeG;omeU;rA;omeA;omeA;rU;omeG;fluA;omeG;rCSup-3′

172
5′-HO;rG;fluC;fluU;rC;omeA;fluU;omeU;omeU;rA;fluC;fluC;rA;omeG;
7802.86
294

omeC;omeG;rU;fluA;omeA;omeA;omeU;omeG;omeA;rG;rCSup-3′

173
5′-HO;rG;omeC;rU;omeC;fluA;fluU;fluU;omeU;rA;fluC;fluC;omeA;omeG;
7808.86
295

fluC;fluG;omeU;omeA;fluA;omeA;omeU;omeG;rA;fluG;omeCSup-3′

174
5′-HO;rG;omeC;rU;omeC;rA;omeU;omeU;rU;rA;omeC;rC;omeA;rG;
7809.00
296

omeC;fluG;rU;omeA;fluA;omeA;omeU;rG;rA;omeG;omeCSup-3′

175
5′-HO;rG;rC;rU;fluC;omeA;omeU;rU;rU;omeA;omeC;rC;omeA;omeG;
7810.94
297

omeC;rG;rU;fluA;omeA;omeA;fluU;omeG;rA;omeG;omeCSup-3′

176
5′-HO;rG;omeC;omeU;rC;omeA;omeU;omeU;rU;omeA;omeC;rC;omeA;
7812.88
298

omeG;omeC;fluG;fluU;rA;rA;rA;omeU;omeG;fluA;fluG;rCSup-3′

177
5′-HO;rG;fluC;omeU;omeC;fluA;rU;rU;omeU;omeA;rC;omeC;rA;omeG;
7814.93
299

omeC;omeG;omeU;rA;fluA;fluA;omeU;rG;omeA;fluG;omeCSup-3′

178
5′-HO;rG;omeC;omeU;omeC;fluA;rU;fluU;omeU;omeA;omeC;omeC;
7816.94
300

omeA;omeG;omeC;rG;fluU;fluA;rA;fluA;rU;omeG;omeA;rG;fluCSup-3′

179
5′-HO;rG;dC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;omeC;
7686.65
301

rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

180
5′-HO;rG;omeC;dU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7700.68
302

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

181
5′-HO;rG;omeC;rU;dC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7700.68
303

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

182
5′-HO;rG;omeC;rU;rC;rA;dU;rU;rU;rA;fluC;rC;rA;fluG;
7698.68
304

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

183
5′-HO;rG;omeC;rU;rC;rA;fluU;dU;rU;rA;fluC;rC;rA;fluG;
7700.68
305

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

184
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;dU;rA;fluC;rC;rA;fluG;
7700.68
306

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

185
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;dC;rC;rA;fluG;
7698.68
307

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

186
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;dC;rA;fluG;
7700.68
308

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

187
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;dA;fluG;
7700.68
309

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

188
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;dG;
7698.68
310

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

189
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7686.65
311

dC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

190
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7700.68
312

omeC;dG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

191
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7698.68
313

omeC;rG;dU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

192
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7686.65
314

omeC;rG;fluU;dA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

193
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7698.68
315

omeC;rG;fluU;omeA;dA;omeA;rU;rG;rA;rG;omeCSup-3′

194
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7686.65
316

omeC;rG;fluU;omeA;fluA;dA;rU;rG;rA;rG;omeCSup-3′

195
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7700.68
317

omeC;rG;fluU;omeA;fluA;omeA;dU;rG;rA;rG;omeCSup-3′

196
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7700.68
318

omeC;rG;fluU;omeA;fluA;omeA;rU;dG;rA;rG;omeCSup-3′

197
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;omeC;rG;
7700.68
319

fluU;omeA;fluA;omeA;rU;rG;dA;rG;omeCSup-3′

198
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7700.68
320

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;dG;omeCSup-3′

199
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7686.65
321

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;dCSup-3′

200
5′-HO;rG;faraC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7704.64
322

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

201
5′-HO;rG;omeC;faraU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7718.67
323

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

202
5′-HO;rG;omeC;rU;faraC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7718.67
324

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

203
5′-HO;rG;omeC;rU;rC;rA;faraU;rU;rU;rA;fluC;rC;rA;fluG;
7716.67
325

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

204
5′-HO;rG;omeC;rU;rC;rA;fluU;faraU;rU;rA;fluC;rC;rA;fluG;
7718.67
326

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

205
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;faraU;rA;fluC;rC;rA;fluG;
7718.67
327

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

206
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;faraC;rC;rA;fluG;
7716.68
328

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

207
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;faraC;rA;fluG;
7718.67
329

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

208
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7704.64
330

faraC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

209
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7716.67
331

omeC;rG;faraU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

210
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7718.67
332

omeC;rG;fluU;omeA;fluA;omeA;faraU;rG;rA;rG;omeCSup-3′

211
5′-HO;rG;dC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7728.73
333

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

212
5′-HO;rG;rC;dU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7714.70
334

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

213
5′-HO;rG;rC;omeU;dC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7728.73
335

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

214
5′-HO;rG;rC;omeU;rC;fluA;dU;omeU;rU;rA;rC;omeC;rA;fluG;
7728.73
336

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

215
5′-HO;rG;rC;omeU;rC;fluA;rU;dU;rU;rA;rC;omeC;rA;fluG;
7714.70
337

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

216
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;dU;rA;rC;omeC;rA;fluG;
7728.73
338

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

217
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;dC;omeC;rA;fluG;
7728.73
339

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

218
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;dC;rA;fluG;
7714.70
340

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

219
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;dA;fluG;
7728.73
341

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

220
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;dG;
7726.74
342

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

221
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7714.70
343

dC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

222
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7728.73
344

omeC;dG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

223
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7714.70
345

omeC;rG;dU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

224
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7714.70
346

omeC;rG;omeU;dA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

225
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7726.74
347

omeC;rG;omeU;omeA;dA;fluA;rU;rG;omeA;rG;fluCSup-3′

226
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7726.74
348

omeC;rG;omeU;omeA;fluA;dA;rU;rG;omeA;rG;fluCSup-3′

227
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7728.73
349

omeC;rG;omeU;omeA;fluA;fluA;dU;rG;omeA;rG;fluCSup-3′

228
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7728.73
350

omeC;rG;omeU;omeA;fluA;fluA;rU;dG;omeA;rG;fluCSup-3′

229
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7714.70
351

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;dA;rG;fluCSup-3′

230
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7728.73
352

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;dG;fluCSup-3′

231
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7726.74
353

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;dCSup-3′

232
5′-HO;rG;faraC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7746.72
354

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

233
5′-HO;rG;rC;faraU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7732.69
355

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

234
5′-HO;rG;rC;omeU;faraC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7746.72
356

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

235
5′-HO;rG;rC;omeU;rC;fluA;faraU;omeU;rU;rA;rC;omeC;rA;fluG;
7746.72
357

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

236
5′-HO;rG;rC;omeU;rC;fluA;rU;faraU;rU;rA;rC;omeC;rA;fluG;
7732.69
358

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

237
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;faraU;rA;rC;omeC;rA;fluG;
7746.72
359

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

238
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;faraC;omeC;rA;fluG;
7746.72
360

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

239
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;faraC;rA;fluG;
7732.69
361

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

240
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;faraA;fluG;
7746.72
362

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

241
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7732.69
363

faraC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

242
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7732.69
364

omeC;rG;faraU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

243
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7746.72
365

omeC;rG;omeU;omeA;fluA;fluA;faraU;rG;omeA;rG;fluCSup-3′

244
5′-HO;rG;omeC;fluU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7704.64
366

omeC;rG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′

245
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;fluC;rA;fluG;
7704.64
367

omeC;rG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′

246
5′-HO;rG;omeC;fluU;rC;rA;fluU;rU;rU;rA;fluC;fluC;rA;fluG;
7706.63
368

omeC;rG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′

247
5′-HO;rG;omeC;rU;rC;rA;fluU;fluU;rU;rA;fluC;rC;rA;fluG;
7704.64
369

omeC;rG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′

248
5′-HO;rG;omeC;fluU;rC;rA;fluU;fluU;rU;rA;fluC;rC;rA;fluG;
7706.63
370

omeC;rG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′

249
5′-HO;rG;omeC;rU;rC;rA;fluU;fluU;rU;rA;fluC;fluC;rA;fluG;
7706.63
371

omeC;rG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′

250
5′-HO;rG;omeC;fluU;rC;rA;fluU;fluU;rU;rA;fluC;fluC;rA;fluG;
7708.62
372

omeC;rG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′

251
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;omeC;
7716.67
373

omeG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′

252
5′-HO;rG;omeC;fluU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7718.67
374

omeC;omeG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′

253
5′-HO;rG;lna5meC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7728.69
375

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

254
5′-HO;rG;omeC;lna5MeU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7742.71
376

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

255
5′-HO;rG;omeC;rU;rC;lnaA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7728.69
377

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

256
5′-HO;rG;omeC;rU;rC;rA;lna5MeU;rU;rU;rA;fluC;rC;rA;fluG;
7740.72
378

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

257
5′-HO;rG;omeC;rU;rC;rA;fluU;lna5MeU;rU;rA;fluC;rC;rA;fluG;
7742.71
379

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

258
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;lna5MeU;rA;fluC;rC;rA;fluG;
7742.71
380

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

259
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;lnaA;fluC;rC;rA;fluG;
7728.69
381

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

260
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;lna5meC;rC;rA;fluG;
7740.72
382

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

261
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;lna5meC;rA;fluG;
7742.71
383

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

262
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;lnaA;fluG;
7728.69
384

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

263
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;lnaG;
7726.69
385

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

264
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7728.69
386

lna5meC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

265
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7728.69
387

omeC;lnaG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

266
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7740.72
388

omeC;rG;lna5MeU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

267
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7714.66
389

omeC;rG;fluU;lnaA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

268
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7726.69
390

omeC;rG;fluU;omeA;lnaA;omeA;rU;rG;rA;rG;omeCSup-3′

269
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7714.66
391

omeC;rG;fluU;omeA;fluA;lnaA;rU;rG;rA;rG;omeCSup-3′

270
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7742.71
392

omeC;rG;fluU;omeA;fluA;omeA;lna5MeU;rG;rA;rG;omeCSup-3′

271
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7728.69
393

omeC;rG;fluU;omeA;fluA;omeA;rU;lnaG;rA;rG;omeCSup-3′

272
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7728.69
394

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;lnaA;rG;omeCSup-3′

273
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7728.69
395

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;lnaG;omeCSup-3′

274
5′-HO;rG;25C;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7702.65
396

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

275
5′-HO;rG;omeC;25U;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7716.67
397

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

276
5′-HO;rG;omeC;rU;25C;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7716.68
398

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

277
5′-HO;rG;omeC;rU;rC;25A;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7716.67
399

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

278
5′-HO;rG;omeC;rU;rC;rA;25U;rU;rU;rA;fluC;rC;rA;fluG;omeC;
7714.68
400

rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

279
5′-HO;rG;omeC;rU;rC;rA;fluU;25U;rU;rA;fluC;rC;rA;fluG;
7716.67
401

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

280
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;25U;rA;fluC;rC;rA;fluG;
7716.67
402

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

281
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;25A;fluC;rC;rA;fluG;
7716.67
403

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

282
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;25C;rC;rA;fluG;omeC;
7714.69
404

rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

283
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;25C;rA;fluG;
7716.68
405

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

284
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;25A;fluG;
7716.67
406

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

285
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;25G;omeC;
7714.68
407

rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

286
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;25C;
7702.65
408

rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

287
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7714.68
409

omeC;rG;25U;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

288
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7702.65
410

omeC;rG;fluU;25A;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

289
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7714.68
411

omeC;rG;fluU;omeA;25A;omeA;rU;rG;rA;rG;omeCSup-3′

290
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7702.65
412

omeC;rG;fluU;omeA;fluA;25A;rU;rG;rA;rG;omeCSup-3′

291
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7716.67
413

omeC;rG;fluU;omeA;fluA;omeA;25U;rG;rA;rG;omeCSup-3′

292
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7716.67
414

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;25A;rG;omeCSup-3′

293
5′-HO;rG;lna5meC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7770.77
415

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

294
5′-HO;rG;rC;lna5MeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7756.74
416

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

295
5′-HO;rG;rC;omeU;lna5meC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7770.77
417

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

296
5′-HO;rG;rC;omeU;rC;lnaA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7754.75
418

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

297
5′-HO;rG;rC;omeU;rC;fluA;lna5MeU;omeU;rU;rA;rC;omeC;rA;fluG;
7770.77
419

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

298
5′-HO;rG;rC;omeU;rC;fluA;rU;lna5MeU;rU;rA;rC;omeC;rA;fluG;
7756.74
420

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

299
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;lna5MeU;rA;rC;omeC;rA;fluG;
7770.77
421

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

300
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;lnaA;rC;omeC;rA;fluG;
7756.74
422

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

301
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;lna5meC;omeC;rA;fluG;
7770.77
423

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

302
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;lna5meC;rA;fluG;
7756.74
424

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

303
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;lnaA;fluG;
7756.74
425

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

304
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;lnaG;
7754.75
426

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

305
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7756.74
427

omeC;lnaG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

306
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7756.74
428

omeC;rG;lna5MeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

307
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7742.71
429

omeC;rG;omeU;lnaA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

308
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7754.75
430

omeC;rG;omeU;omeA;lnaA;fluA;rU;rG;omeA;rG;fluCSup-3′

309
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7754.75
431

omeC;rG;omeU;omeA;fluA;lnaA;rU;rG;omeA;rG;fluCSup-3′

310
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7770.77
432

omeC;rG;omeU;omeA;fluA;fluA;lna5MeU;rG;omeA;rG;fluCSup-3′

311
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7756.74
433

omeC;rG;omeU;omeA;fluA;fluA;rU;lnaG;omeA;rG;fluCSup-3′

312
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7742.71
434

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;lnaA;rG;fluCSup-3′

313
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7756.74
435

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;lnaG;fluCSup-3′

314
5′-HO;rG;25C;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7744.73
436

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

315
5′-HO;rG;rC;25U;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7730.70
437

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

316
5′-HO;rG;rC;omeU;25C;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7744.73
438

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

317
5′-HO;rG;rC;omeU;rC;25A;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7742.74
439

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

318
5′-HO;rG;rC;omeU;rC;fluA;25U;omeU;rU;rA;rC;omeC;rA;fluG;
7744.73
440

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

319
5′-HO;rG;rC;omeU;rC;fluA;rU;25U;rU;rA;rC;omeC;rA;fluG;
7730.70
441

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

320
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;25U;rA;rC;omeC;rA;fluG;
7744.73
442

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

321
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;25A;rC;omeC;rA;fluG;
7744.73
443

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

322
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;25C;omeC;rA;fluG;
7744.73
444

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

323
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;25C;rA;fluG;
7730.71
445

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

324
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;25A;fluG;
7744.73
446

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

325
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;25G;
7742.74
447

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

326
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7730.71
448

25C;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

327
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7744.73
449

omeC;25G;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

328
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7730.70
450

omeC;rG;25U;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

329
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7730.70
451

omeC;rG;omeU;25A;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

330
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7742.74
452

omeC;rG;omeU;omeA;25A;fluA;rU;rG;omeA;rG;fluCSup-3′

331
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7742.74
453

omeC;rG;omeU;omeA;fluA;25A;rU;rG;omeA;rG;fluCSup-3′

332
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7744.73
454

omeC;rG;omeU;omeA;fluA;fluA;25U;rG;omeA;rG;fluCSup-3′

333
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7730.70
455

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;25A;rG;fluCSup-3′

334
5′-HO;rG;rC;omeU;rC;fluA;fluU;omeU;rU;rA;rC;omeC;rA;fluG;
7756.76
456

omeC;rG;omeU;omeA;rA;fluA;rU;rG;omeA;rG;omeCSup-3′

335
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;fluC;omeC;rA;fluG;
7756.76
457

omeC;rG;omeU;omeA;rA;fluA;rU;rG;omeA;rG;omeCSup-3′

336
5′-HO;rG;rC;omeU;rC;fluA;fluU;omeU;rU;rA;fluC;omeC;rA;fluG;
7758.75
458

omeC;rG;omeU;omeA;rA;fluA;rU;rG;omeA;rG;omeCSup-3′

337
5′-HO;rG;omeC;rU;rC;r2AP;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7716.68
2

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

338
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;r2AP;fluG;
7716.68
3

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

339
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7702.65
4

omeC;rG;fluU;r2AP;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

340
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7714.68
5

omeC;rG;fluU;omeA;r2AP;omeA;rU;rG;rA;rG;omeCSup-3′

341
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7702.65
6

omeC;rG;fluU;omeA;fluA;r2AP;rU;rG;rA;rG;omeCSup-3′

342
5′-HO;rG;omeC;rU;rC;moeA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7774.75
459

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

343
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;moeA;fluC;rC;rA;fluG;
7774.75
460

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

344
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;moeA;fluG;
7774.75
461

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

345
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;moeG;
7772.76
462

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

346
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7774.75
463

omeC;moeG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

347
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;omeC;
7760.73
464

rG;fluU;moeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

348
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7772.76
465

omeC;rG;fluU;omeA;moeA;omeA;rU;rG;rA;rG;omeCSup-3′

349
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7760.73
466

omeC;rG;fluU;omeA;fluA;moeA;rU;rG;rA;rG;omeCSup-3′

350
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7774.75
467

omeC;rG;fluU;omeA;fluA;omeA;rU;moeG;rA;rG;omeCSup-3′

351
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7774.75
468

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;moeA;rG;omeCSup-3′

352
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7774.75
469

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;moeG;omeCSup-3′

353
5′-HO;rG;rC;omeU;rC;r2AP;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7742.74
7

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

354
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;r2AP;rC;omeC;rA;fluG;
7744.73
8

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

355
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;r2AP;fluG;
7744.73
9

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

356
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7730.70
10

omeC;rG;omeU;r2AP;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

357
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7742.74
11

omeC;rG;omeU;omeA;r2AP;fluA;rU;rG;omeA;rG;fluCSup-3′

358
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7742.74
12

omeC;rG;omeU;omeA;fluA;r2AP;rU;rG;omeA;rG;fluCSup-3′

359
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7730.70
13

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;r2AP;rG;fluCSup-3′

360
5′-HO;rG;rC;omeU;rC;moeA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7800.82
470

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

361
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;moeA;rC;omeC;rA;fluG;
7802.81
471

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

362
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;moeA;fluG;
7802.81
472

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

363
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;moeG;
7800.82
473

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

364
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7802.81
474

omeC;moeG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

365
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7788.78
475

omeC;rG;omeU;moeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

366
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7800.82
476

omeC;rG;omeU;omeA;moeA;fluA;rU;rG;omeA;rG;fluCSup-3′

367
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7800.82
477

omeC;rG;omeU;omeA;fluA;moeA;rU;rG;omeA;rG;fluCSup-3′

368
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7802.81
478

omeC;rG;omeU;omeA;fluA;fluA;rU;moeG;omeA;rG;fluCSup-3′

369
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7788.78
479

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;moeA;rG;fluCSup-3′

370
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7802.81
480

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;moeG;fluCSup-3′

371
5′-HO;rG;omeC;fluU;rC;rA;fluU;fluU;rU;rA;fluC;rC;rA;fluG;
7720.66
481

omeC;omeG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′

372
5′-HO;rG;omeC;rU;rC;rA;fluU;fluU;rU;rA;fluC;fluC;rA;fluG;
7720.66
482

omeC;omeG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′

373
5′-HO;rG;omeC;fluU;rC;rA;fluU;fluU;rU;rA;fluC;fluC;rA;fluG;
7722.65
483

omeC;omeG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′

374
5′-HO;rG;moe5meC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7774.75
484

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

375
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;moe5meC
7774.75
485

rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′;

376
5′-HO;rG;omeC;rU;rC;rA;moe-5-MeU;rU;rU;rA;fluC;rC;rA;fluG;
7786.79
486

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

377
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;moe5meC;rC;rA;fluG;
7786.79
487

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

378
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7786.79
488

omeC;rG;moe5MeU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

379
5′-HO;rG;omeC;moe5MeU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7788.78
489

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

380
5′-HO;rG;omeC;rU;moe5meC;rA;fluU;rU;rU;rA;fluC;rC;rA;
7788.78
490

fluG;omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

381
5′-HO;rG;omeC;rU;rC;rA;fluU;moe5MeU;rU;rA;fluC;rC;rA;fluG;
7788.78
491

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

382
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;moe5MeU;rA;fluC;rC;rA;fluG;
7788.78
492

omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

383
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;moe5meC;rA;
7788.78
493

fluG;omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′

384
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;
7788.78
494

omeC;rG;fluU;omeA;fluA;omeA;moe5MeU;rG;rA;rG;omeCSup-3′

385
5′-HO;rG;rC;moe5MeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7802.81
495

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

386
5′-HO;rG;rC;omeU;rC;fluA;rU;moe5MeU;rU;rA;rC;omeC;rA;fluG;
7802.81
496

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

387
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;moe5meC;rA;
7802.81
497

fluG;omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

388
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7802.81
498

moe5meC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

389
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7802.81
499

omeC;rG;moe5MeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

390
5′-HO;rG;moe5meC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7816.83
500

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

391
5′-HO;rG;rC;omeU;moe5meC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7816.83
501

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

392
5′-HO;rG;rC;omeU;rC;fluA;moe5MeU;omeU;rU;rA;rC;omeC;rA;fluG;
7816.83
502

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

393
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;moe5MeU;rA;rC;omeC;rA;fluG;
7816.83
503

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

394
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;moe5meC;omeC;rA;fluG;
7816.83
504

omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′

395
5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG;
7816.83
505

omeC;rG;omeU;omeA;fluA;fluA;moe5MeU;rG;omeA;rG;fluCSup-3′

396
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7797.00
506

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

397
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7873.16
507

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

398
5′-HO;rG;omeC;rU;rC;rI;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7797.99
508

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

399
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rI;fluC;rC;rAs;fluG;
7797.99
509

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

400
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rIs;fluG;
7797.99
510

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

401
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7783.96
511

omeC;rG;fluU;rI;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

402
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7796.00
512

omeC;rG;fluU;omeA;rI;omeA;rUs;rG;rAs;rGs;omeCSup-3′

403
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7783.96
513

omeC;rG;fluU;omeA;fluA;rI;rUs;rG;rAs;rGs;omeCSup-3′

404
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7797.99
514

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rIs;rGs;omeCSup-3′

405
5′-HO;rG;d5propC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7805.03
14

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

406
5′-HO;rG;omeC;d5propU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7819.05
15

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

407
5′-HO;rG;omeC;rU;d5propC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7819.05
16

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

408
5′-HO;rG;omeC;rU;rC;rA;d5propU;rU;rUs;rA;fluC;rC;rAs;fluG;
7817.06
17

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

409
5′-HO;rG;omeC;rU;rC;rA;fluU;d5propU;rUs;rA;fluC;rC;rAs;fluG;
7819.05
18

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

410
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;d5propUs;rA;fluC;rC;rAs;fluG;
7819.05
19

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

411
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;d5propC;rC;rAs;fluG;
7817.06
20

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

412
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;d5propC;rAs;fluG;
7819.05
21

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

413
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7805.03
22

d5propC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

414
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7817.06
23

rG;d5propU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

415
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7819.05
24

rG;fluU;omeA;fluA;omeA;d5propUs;rG;rAs;rGs;omeCSup-3′

416
5′-HO;rG;omeC;rU;rC;r7dz8azA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7797.01
25

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

417
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;r7dz8azA;fluC;rC;rAs;fluG;
7797.01
26

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

418
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;r7dz8azAs;fluG;
7797.00
27

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

419
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7782.98
28

rG;fluU;r7dz8azA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

420
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7795.01
29

rG;fluU;omeA;r7dz8azA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

421
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7782.98
30

rG;fluU;omeA;fluA;r7dz8azA;rUs;rG;rAs;rGs;omeCSup-3′

422
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7797.00
31

rG;fluU;omeA;fluA;omeA;rUs;rG;r7dz8azAs;rGs;omeCSup-3′

423
5′-HO;rG;omeC;rU;rC;d2aminoA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7796.02
32

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

424
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;d2aminoA;fluC;rC;rAs;fluG;
7796.02
33

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

425
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;d2aminoAs;fluG;
7796.02
34

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

426
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7781.99
35

fluU;d2aminoA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

427
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7794.03
36

fluU;omeA;d2aminoA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

428
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7781.99
37

fluU;omeA;fluA;d2aminoA;rUs;rG;rAs;rGs;omeCSup-3′

429
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7796.02
38

fluU;omeA;fluA;omeA;rUs;rG;d2aminoAs;rGs;omeCSup-3′

430
5′-HO;rG;omeC;rU;rC;fara8azA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7799.98
39

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

431
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;fara8azA;fluC;rC;rAs;fluG;
7799.98
40

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

432
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;fara8azAs;fluG;omeC;
7799.98
41

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

433
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7785.96
42

fluU;fara8azA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

434
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7797.99
43

fluU;omeA;fara8azA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

435
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7785.96
44

fluU;omeA;fluA;fara8azA;rUs;rG;rAs;rGs;omeCSup-3′

436
5′-HO;rG;rC;rUs;fluC;rI;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;
7860.12
515

omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

437
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;rI;omeC;omeC;rAs;omeG;
7872.15
516

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

438
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rIs;omeG;
7874.14
517

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

439
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7874.14
518

omeC;omeG;omeU;rI;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

440
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7872.15
519

omeC;omeG;omeU;rA;rI;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

441
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7872.15
520

omeC;omeG;omeU;rA;fluA;rI;omeUs;fluG;omeAs;rGs;fluCSup-3′

442
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7860.12
521

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;rIs;rGs;fluCSup-3′

443
5′-HO;rG;d5propC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7895.21
45

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

444
5′-HO;rG;rC;d5propUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7895.20
46

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

445
5′-HO;rG;rC;rUs;d5propC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7893.22
47

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

446
5′-HO;rG;rC;rUs;fluC;omeA;d5propU;rU;rUs;fluA;omeC;omeC;rAs;
7893.22
48

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

447
5′-HO;rG;rC;rUs;fluC;omeA;fluU;d5propU;rUs;fluA;omeC;omeC;rAs;
7895.21
49

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

448
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;d5propUs;fluA;omeC;omeC;rAs;
7895.20
50

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

449
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;d5propC;omeC;rAs;omeG;
7881.18
51

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

450
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;d5propC;rAs;omeG;
7881.18
52

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

451
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7881.18
53

d5propC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

452
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7881.18
54

omeC;omeG;d5propU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

453
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7881.18
55

omeC;omeG;omeU;rA;fluA;fluA;d5propUs;fluG;omeAs;rGs;fluCSup-3′

454
5′-HO;rG;rC;rUs;fluC;r7dz8azA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7859.13
56

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

455
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;r7dz8azA;omeC;omeC;rAs;
7871.17
57

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

456
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;r7dz8azAs;
7873.16
58

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

457
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7873.16
59

omeC;omeG;omeU;r7dz8azA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

458
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7871.17
60

omeC;omeG;omeU;rA;r7dz8azA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

459
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7871.17
61

omeC;omeG;omeU;rA;fluA;r7dz8azA;omeUs;fluG;omeAs;rGs;fluCSup-3′

460
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7859.13
62

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;r7dz8azAs;rGs;fluCSup-3′

461
5′-HO;rG;rC;rUs;fluC;d2aminoA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7858.15
63

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

462
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;d2aminoA;omeC;omeC;rAs;
7870.18
64

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

463
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;d2aminoAs;
7872.17
65

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

464
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7872.18
66

omeC;omeG;omeU;d2aminoA;fluA;fluA;omeUs;fluG;omeAs;rGs;

fluCSup-3′

465
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7870.18
67

omeC;omeG;omeU;rA;d2aminoA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

466
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7870.18
68

omeC;omeG;omeU;rA;fluA;d2aminoA;omeUs;fluG;omeAs;rGs;fluCSup-3′

467
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7858.14
69

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;d2aminoAs;rGs;fluCSup-3′

468
5′-HO;rG;rC;rUs;fluC;fara8azA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7862.11
70

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

469
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fara8azA;omeC;omeC;rAs;
7874.15
71

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

470
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;fara8azAs;
7876.13
72

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

471
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7876.14
73

omeC;omeG;omeU;fara8azA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

472
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7874.15
74

omeC;omeG;omeU;rA;fara8azA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

473
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7874.15
75

omeC;omeG;omeU;rA;fluA;fara8azA;omeUs;fluG;omeAs;rGs;fluCSup-3′

474
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7862.11
76

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;fara8azAs;rGs;fluCSup-3′

475
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;fluC;rAs;fluG;omeC;
7798.99
522

omeG;fluU;omeA;rA;fluA;rUs;rG;rAs;rGs;omeCSup-3′

476
5′-HO;rG;omeC;fluU;rC;rA;fluU;rU;rUs;rA;fluC;fluC;rAs;fluG;omeC;
7800.99
523

omeG;fluU;omeA;rA;fluA;rUs;rG;rAs;rGs;omeCSup-3′

477
5′-HO;rG;omeC;rU;rC;rA;fluU;fluU;rUs;rA;fluC;rC;rAs;fluG;
7798.99
524

omeC;omeG;fluU;omeA;rA;fluA;rUs;rG;rAs;rGs;omeCSup-3′

478
5′-HO;rG;omeC;rU;rC;fara7dzA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7798.01
77

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

479
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;fara7dzA;fluC;rC;rAs;fluG;
7798.01
78

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

480
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;fara7dzAs;fluG;omeC;
7798.00
79

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

481
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7783.98
80

omeC;rG;fluU;fara7dzA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

482
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7796.02
81

fluU;omeA;fara7dzA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

483
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7783.98
82

omeC;rG;fluU;omeA;fluA;fara7dzA;rUs;rG;rAs;rGs;omeCSup-3′

484
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7798.00
83

fluU;omeA;fluA;omeA;rUs;rG;fara7dzAs;rGs;omeCSup-3′

485
5′-HO;rG;omeC;rU;rC;d7dzA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7780.02
84

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

486
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;d7dzA;fluC;rC;rAs;fluG;
7780.02
85

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

487
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;d7dzAs;fluG;
7780.01
86

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

488
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7765.99
87

omeC;rG;fluU;d7dzA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

489
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7778.03
88

omeC;rG;fluU;omeA;d7dzA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

490
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7765.99
89

omeC;rG;fluU;omeA;fluA;d7dzA;rUs;rG;rAs;rGs;omeCSup-3′

491
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7780.01
90

fluU;omeA;fluA;omeA;rUs;rG;d7dzAs;rGs;omeCSup-3′

492
5′-HO;rG;omeC;rU;rC;r7dzA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7796.02
91

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

493
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;r7dzA;fluC;rC;rAs;fluG;
7796.02
92

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

494
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;r7dzAs;fluG;
7796.01
93

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

495
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7781.99
94

omeC;rG;fluU;r7dzA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

496
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7794.03
95

omeC;rG;fluU;omeA;r7dzA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

497
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7781.99
96

omeC;rG;fluU;omeA;fluA;r7dzA;rUs;rG;rAs;rGs;omeCSup-3′

498
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7796.01
97

fluU;omeA;fluA;omeA;rUs;rG;r7dzAs;rGs;omeCSup-3′

499
5′-HO;rG;rC;rUs;fluC;fara7dzA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7860.14
98

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

500
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fara7dzA;omeC;omeC;rAs;
7872.17
99

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

501
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;fara7dzAs;
7874.16
100

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

502
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7874.16
101

omeC;omeG;omeU;fara7dzA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

503
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7872.17
102

omeC;omeG;omeU;rA;fara7dzA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

504
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7872.17
103

omeC;omeG;omeU;rA;fluA;fara7dzA;omeUs;fluG;omeAs;rGs;fluCSup-3′

505
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7860.13
104

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;fara7dzAs;rGs;fluCSup-3′

506
5′-HO;rG;rC;rUs;fluC;d7dzA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7842.15
105

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

507
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;d7dzA;omeC;omeC;rAs;omeG;
7854.18
106

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

508
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;d7dzAs;
7856.17
107

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

509
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7856.17
108

omeC;omeG;omeU;d7dzA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

510
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7854.18
109

omeC;omeG;omeU;rA;d7dzA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

511
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7854.18
110

omeC;omeG;omeU;rA;fluA;d7dzA;omeUs;fluG;omeAs;rGs;fluCSup-3′

512
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7842.14
111

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;d7dzAs;rGs;fluCSup-3′

513
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;r7dzA;omeC;omeC;rAs;omeG;
7870.18
112

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

514
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;fluC;omeC;rAs;omeG;
7843.11
525

omeC;omeG;omeU;rA;rA;fluA;rUs;rG;omeAs;rGs;fluCSup-3′

515
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7855.15
526

omeC;omeG;omeU;rA;rA;fluA;rUs;rG;omeAs;rGs;fluCSup-3′

516
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;fluC;omeC;rAs;omeG;
7845.10
527

omeC;omeG;omeU;rA;fluA;fluA;rUs;rG;omeAs;rGs;fluCSup-3′

517
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;fluC;omeC;rAs;omeG;
7857.14
528

omeC;omeG;omeU;rA;rA;fluA;omeUs;rG;omeAs;rGs;fluCSup-3′

518
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;fluC;omeC;rAs;omeG;
7845.10
529

omeC;omeG;omeU;rA;rA;fluA;rUs;fluG;omeAs;rGs;fluCSup-3′

519
5′-HO;thiorG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7813.07
530

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

520
5′-HO;rG;omeC;thiorU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7813.06
531

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

521
5′-HO;rG;omeC;rU;thiorC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7813.07
532

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

522
5′-HO;rG;omeC;rU;rC;thiorA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7813.07
533

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

523
5′-HO;rG;omeC;rU;rC;rA;thiorU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7811.07
534

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

524
5′-HO;rG;omeC;rU;rC;rA;fluU;thiorU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7813.06
535

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

525
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;thiorUs;rA;fluC;rC;rAs;fluG;omeC;
7813.06
536

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

526
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;thiorA;fluC;rC;rAs;fluG;omeC;
7813.07
537

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

527
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;thiorC;rC;rAs;fluG;omeC;
7811.07
538

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

528
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;thiorC;rAs;fluG;omeC;
7813.07
539

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

529
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;thiorAs;fluG;omeC;
7813.06
540

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

530
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;thiorG;omeC;
7811.08
541

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

531
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;thiorC;rG;
7799.04
542

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

532
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7813.07
543

thiorG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

533
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7811.07
544

thiorU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

534
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7799.04
545

fluU;thiorA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

535
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7811.08
546

fluU;omeA;thiorA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

536
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7799.04
547

fluU;omeA;fluA;thiorA;rUs;rG;rAs;rGs;omeCSup-3′

537
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7813.06
548

fluU;omeA;fluA;omeA;thiorUs;rG;rAs;rGs;omeCSup-3′

538
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7813.07
549

fluU;omeA;fluA;omeA;rUs;thiorG;rAs;rGs;omeCSup-3′

539
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7813.06
550

fluU;omeA;fluA;omeA;rUs;rG;thiorAs;rGs;omeCSup-3′

540
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7813.06
551

fluU;omeA;fluA;omeA;rUs;rG;rAs;thiorGs;omeCSup-3′

541
5′-HO;rG;cf3C;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7850.97
552

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

542
5′-HO;rG;omeC;cf3U;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7865.00
553

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

543
5′-HO;rG;omeC;rU;cf3C;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7865.00
554

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

544
5′-HO;rG;omeC;rU;rC;rA;cf3U;rU;rUs;rA;fluC;rC;rAs;fluG;
7863.01
555

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

545
5′-HO;rG;omeC;rU;rC;rA;fluU;cf3U;rUs;rA;fluC;rC;rAs;fluG;
7865.00
556

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

546
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;cf3Us;rA;fluC;rC;rAs;fluG;
7865.00
557

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

547
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;cf3C;rC;rAs;fluG;
7863.01
558

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

548
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;cf3C;rAs;fluG;
7865.00
559

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

549
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;cf3As;fluG;
7865.00
560

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

550
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7850.97
561

cf3C;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

551
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7863.01
562

omeC;rG;cf3U;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

552
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7850.97
563

omeC;rG;fluU;cf3A;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

553
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7863.01
564

omeC;rG;fluU;omeA;cf3A;omeA;rUs;rG;rAs;rGs;omeCSup-3′

554
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7865.00
565

omeC;rG;fluU;omeA;fluA;omeA;cf3Us;rG;rAs;rGs;omeCSup-3′

555
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7865.00
566

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;cf3As;rGs;omeCSup-3′

556
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;
7865.00
567

omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;cf3Gs;omeCSup-3′

557
5′-HO;thiorG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7889.22
568

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

558
5′-HO;rG;thiorC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7889.22
569

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

559
5′-HO;rG;rC;thiorUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7889.21
570

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

560
5′-HO;rG;rC;rUs;fluC;omeA;thiorU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7887.23
571

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

561
5′-HO;rG;rC;rUs;fluC;omeA;fluU;thiorU;rUs;fluA;omeC;omeC;rAs;
7889.22
572

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

562
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;thiorUs;fluA;omeC;omeC;rAs;
7889.21
573

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

563
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;thiorA;omeC;omeC;rAs;omeG;
7887.23
574

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

564
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;thiorC;omeC;rAs;omeG;
7875.19
575

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

565
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;thiorC;rAs;omeG;
7875.19
576

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

566
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;thiorAs;
7889.22
577

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

567
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;thiorG;
7875.19
578

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

568
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7875.19
579

thiorC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

569
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7875.19
580

omeC;thiorG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

570
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7875.19
581

omeC;omeG;thiorU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

571
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7889.22
582

omeC;omeG;omeU;thiorA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

572
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7887.23
583

omeC;omeG;omeU;rA;thiorA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

573
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7887.23
584

omeC;omeG;omeU;rA;fluA;thiorA;omeUs;fluG;omeAs;rGs;fluCSup-3′

574
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7875.19
585

omeC;omeG;omeU;rA;fluA;fluA;thiorUs;fluG;omeAs;rGs;fluCSup-3′

575
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7887.23
586

omeC;omeG;omeU;rA;fluA;fluA;omeUs;thiorG;omeAs;rGs;fluCSup-3′

576
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7875.19
587

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;thiorAs;rGs;fluCSup-3′

577
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7889.22
588

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;thiorGs;fluCSup-3′

578
5′-HO;rG;cf3C;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7941.16
589

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

579
5′-HO;rG;rC;cf3Us;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7941.16
590

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

580
5′-HO;rG;rC;rUs;cf3C;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7939.16
591

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

581
5′-HO;rG;rC;rUs;fluC;omeA;cf3U;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7939.16
592

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

582
5′-HO;rG;rC;rUs;fluC;omeA;fluU;cf3U;rUs;fluA;omeC;omeC;rAs;omeG;
7941.16
593

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

583
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;cf3Us;fluA;omeC;omeC;rAs;omeG;
7941.16
594

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

584
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;cf3A;omeC;omeC;rAs;omeG;
7939.16
595

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

585
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;cf3C;omeC;rAs;omeG;
7927.13
596

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

586
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;cf3C;rAs;omeG;
7927.13
597

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

587
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;cf3As;omeG;
7941.16
598

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

588
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7927.13
599

cf3C;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

589
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7927.13
600

omeC;omeG;cf3U;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

590
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7939.16
601

omeC;omeG;omeU;rA;cf3A;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

591
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7939.16
602

omeC;omeG;omeU;rA;fluA;cf3A;omeUs;fluG;omeAs;rGs;fluCSup-3′

592
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7927.13
603

omeC;omeG;omeU;rA;fluA;fluA;cf3Us;fluG;omeAs;rGs;fluCSup-3′

593
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7927.13
604

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;cf3As;rGs;fluCSup-3′

594
5′-HO;4S8azG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7814.05
113

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

595
5′-HO;rG;araC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7782.98
605

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

596
5′-HO;rG;omeC;araU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7797.00
606

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

597
5′-HO;rG;omeC;rU;araC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7797.00
607

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

598
5′-HO;rG;omeC;rU;rC;araA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7797.00
608

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

599
5′-HO;rG;omeC;rU;rC;rA;araU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7795.01
609

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

600
5′-HO;rG;omeC;rU;rC;rA;fluU;araU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7797.00
610

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

601
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;araUs;rA;fluC;rC;rAs;fluG;omeC;
7797.00
611

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

602
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;araA;fluC;rC;rAs;fluG;omeC;
7797.00
612

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

603
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;araC;rC;rAs;fluG;omeC;rG;
7795.01
613

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

604
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;araC;rAs;fluG;omeC;
7797.00
614

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

605
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;araAs;fluG;omeC;
7797.00
615

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

606
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;4S8azG;omeC;
7812.06
114

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

607
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;araC;rG;
7782.98
616

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

608
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7814.05
115

4S8azG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

609
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7795.01
617

araU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

610
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7782.98
618

fluU;araA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

611
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7795.01
619

fluU;omeA;araA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

612
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7782.98
620

fluU;omeA;fluA;araA;rUs;rG;rAs;rGs;omeCSup-3′

613
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7797.00
621

fluU;omeA;fluA;omeA;araUs;rG;rAs;rGs;omeCSup-3′

614
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7814.05
116

fluU;omeA;fluA;omeA;rUs;4S8azG;rAs;rGs;omeCSup-3′

615
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7797.00
622

fluU;omeA;fluA;omeA;rUs;rG;araAs;rGs;omeCSup-3′

616
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7814.05
117

fluU;omeA;fluA;omeA;rUs;rG;rAs;4S8azGs;omeCSup-3′

617
5′-HO;rG;omeC;rU;rC;rA;rxyloU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7795.01
623

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

618
5′-HO;rG;omeC;rU;rC;rA;fluU;rxyloU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7797.00
624

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

619
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rxyloUs;rA;fluC;rC;rAs;fluG;omeC;
7797.00
625

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

620
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;rxyloC;rC;rAs;fluG;omeC;
7795.01
626

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

621
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rxyloC;rAs;fluG;omeC;
7797.00
627

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

622
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rxyloAs;fluG;omeC;
7797.00
628

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

623
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;rxyloG;omeC;
7795.01
629

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

624
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;rxyloC;rG;
7782.98
630

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

625
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rxyloG;
7797.00
631

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

626
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7795.01
632

rxyloU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

627
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7782.98
633

fluU;rxyloA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

628
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7795.01
634

fluU;omeA;rxyloA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

629
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7782.98
635

fluU;omeA;fluA;rxyloA;rUs;rG;rAs;rGs;omeCSup-3′

630
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7797.00
636

fluU;omeA;fluA;omeA;rxyloUs;rG;rAs;rGs;omeCSup-3′

631
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7797.00
637

fluU;omeA;fluA;omeA;rUs;rxyloG;rAs;rGs;omeCSup-3′

632
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7797.00
638

fluU;omeA;fluA;omeA;rUs;rG;rxyloAs;rGs;omeCSup-3′

633
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7797.00
639

fluU;omeA;fluA;omeA;rUs;rG;rAs;rxyloGs;omeCSup-3′

634
5′-HO;4S8azG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7890.21
118

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

635
5′-HO;rG;araC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7873.16
640

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

636
5′-HO;rG;rC;araUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7873.16
641

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

637
5′-HO;rG;rC;rUs;araC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7871.17
642

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

638
5′-HO;rG;rC;rUs;fluC;araA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7859.13
643

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

639
5′-HO;rG;rC;rUs;fluC;omeA;araU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7871.17
644

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

640
5′-HO;rG;rC;rUs;fluC;omeA;fluU;araU;rUs;fluA;omeC;omeC;rAs;omeG;
7873.16
645

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

641
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;araUs;fluA;omeC;omeC;rAs;omeG;
7873.16
646

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

642
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;araA;omeC;omeC;rAs;omeG;
7871.17
647

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

643
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;araC;omeC;rAs;omeG;
7859.13
648

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

644
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;araC;rAs;omeG;
7859.13
649

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

645
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;araAs;omeG;
7873.16
650

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

646
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;4S8azG;
7876.18
119

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

647
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7859.13
651

araC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

648
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7876.18
120

omeC;4S8azG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

649
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7859.13
652

omeC;omeG;araU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

650
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7873.16
653

omeC;omeG;omeU;araA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

651
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7871.17
654

omeC;omeG;omeU;rA;araA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

652
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7871.17
655

omeC;omeG;omeU;rA;fluA;araA;omeUs;fluG;omeAs;rGs;fluCSup-3′

653
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7859.13
656

omeC;omeG;omeU;rA;fluA;fluA;araUs;fluG;omeAs;rGs;fluCSup-3′

654
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7888.22
121

omeC;omeG;omeU;rA;fluA;fluA;omeUs;4S8azG;omeAs;rGs;fluCSup-3′

655
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7859.13
657

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;araAs;rGs;fluCSup-3′

656
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7890.20
122

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;4S8azGs;fluCSup-3′

657
5′-HO;rG;rC;rUs;fluC;omeA;rxyloU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7871.17
658

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

658
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rxyloU;rUs;fluA;omeC;omeC;rAs;omeG;
7873.16
659

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

659
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rxyloUs;fluA;omeC;omeC;rAs;omeG;
7873.16
660

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

660
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;rxyloC;rAs;omeG;
7859.13
661

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

661
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rxyloAs;
7873.16
662

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

662
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;rxyloG;
7859.13
663

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

663
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7859.13
664

rxyloC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

664
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7859.13
665

omeC;rxyloG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

665
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7859.13
666

omeC;omeG;rxyloU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

666
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7873.16
667

omeC;omeG;omeU;rxyloA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

667
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7871.17
668

omeC;omeG;omeU;rA;rxyloA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

668
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7871.17
669

omeC;omeG;omeU;rA;fluA;rxyloA;omeUs;fluG;omeAs;rGs;fluCSup-3′

669
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7859.13
670

omeC;omeG;omeU;rA;fluA;fluA;rxyloUs;fluG;omeAs;rGs;fluCSup-3′

670
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7871.17
671

omeC;omeG;omeU;rA;fluA;fluA;omeUs;rxyloG;omeAs;rGs;fluCSup-3′

671
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7859.13
672

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;rxyloAs;rGs;fluCSup-3′

672
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7873.16
673

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rxyloGs;fluCSup-3′

673
5′-HO;amiG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7796.02
674

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

674
5′-HO;rG;amiC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7781.99
675

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

675
5′-HO;rG;omeC;amiU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7796.02
676

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

676
5′-HO;rG;omeC;rU;rC;amiA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7796.02
677

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

677
5′-HO;rG;omeC;rU;rC;rA;amiU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7794.03
678

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

678
5′-HO;rG;omeC;rU;rC;rA;fluU;amiU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7796.02
679

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

679
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;amiUs;rA;fluC;rC;rAs;fluG;omeC;
7796.02
680

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

680
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;amiC;rC;rAs;fluG;omeC;rG;
7794.03
681

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

681
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;amiAs;fluG;omeC;
7796.02
682

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

682
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;amiG;omeC;rG;
7794.03
683

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

683
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;amiC;rG;
7781.99
684

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

684
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7794.03
685

amiU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

685
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7794.03
686

fluU;omeA;amiA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

686
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7781.99
687

fluU;omeA;fluA;amiA;rUs;rG;rAs;rGs;omeCSup-3′

687
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7796.02
688

fluU;omeA;fluA;omeA;amiUs;rG;rAs;rGs;omeCSup-3′

688
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7796.02
689

fluU;omeA;fluA;omeA;rUs;amiG;rAs;rGs;omeCSup-3′

689
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7796.02
690

fluU;omeA;fluA;omeA;rUs;rG;amiAs;rGs;omeCSup-3′

690
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7796.02
691

fluU;omeA;fluA;omeA;rUs;rG;rAs;amiGs;omeCSup-3′

691
5′-HO;rG;unaC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7784.99
692

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

692
5′-HO;rG;omeC;unaU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7799.02
693

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

693
5′-HO;rG;omeC;rU;unaC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7799.02
694

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

694
5′-HO;rG;omeC;rU;rC;unaA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7799.02
695

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

695
5′-HO;rG;omeC;rU;rC;rA;unaU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7797.03
696

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

696
5′-HO;rG;omeC;rU;rC;rA;fluU;unaU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7799.02
697

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

697
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;unaUs;rA;fluC;rC;rAs;fluG;omeC;
7799.02
698

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

698
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;unaA;fluC;rC;rAs;fluG;omeC;
7799.02
699

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

699
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;unaC;rC;rAs;fluG;omeC;rG;
7797.03
700

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

700
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;unaC;rAs;fluG;omeC;
7799.02
701

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

701
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;unaAs;fluG;omeC;
7799.02
702

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

702
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;unaG;omeC;rG;
7797.03
703

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

703
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;unaC;rG;
7784.99
704

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

704
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;unaG;
7799.02
705

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

705
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7797.03
706

unaU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

706
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7784.99
707

fluU;unaA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

707
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7797.03
708

fluU;omeA;unaA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

708
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7784.99
709

fluU;omeA;fluA;unaA;rUs;rG;rAs;rGs;omeCSup-3′

709
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7799.02
710

fluU;omeA;fluA;omeA;unaUs;rG;rAs;rGs;omeCSup-3′

710
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7799.02
711

fluU;omeA;fluA;omeA;rUs;unaG;rAs;rGs;omeCSup-3′

711
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7799.02
712

fluU;omeA;fluA;omeA;rUs;rG;unaAs;rGs;omeCSup-3′

712
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7799.02
713

fluU;omeA;fluA;omeA;rUs;rG;rAs;unaGs;omeCSup-3′

713
5′-HO;amiG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7872.17
714

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

714
5′-HO;rG;amiC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7872.17
715

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

715
5′-HO;rG;rC;amiUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7872.17
716

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

716
5′-HO;rG;rC;rUs;amiC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7870.18
717

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

717
5′-HO;rG;rC;rUs;fluC;amiA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7858.15
718

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

718
5′-HO;rG;rC;rUs;fluC;omeA;amiU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7870.18
719

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

719
5′-HO;rG;rC;rUs;fluC;omeA;fluU;amiU;rUs;fluA;omeC;omeC;rAs;omeG;
7872.17
720

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

720
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;amiUs;fluA;omeC;omeC;rAs;omeG;
7872.17
721

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

721
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;amiC;omeC;rAs;omeG;
7858.15
722

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

722
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;amiC;rAs;omeG;
7858.15
723

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

723
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;amiAs;omeG;
7872.17
724

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

724
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;amiG;
7858.15
725

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

725
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7858.15
726

amiC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

726
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7858.15
727

omeC;amiG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

727
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7858.15
728

omeC;omeG;amiU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

728
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7872.17
729

omeC;omeG;omeU;amiA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

729
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7870.18
730

omeC;omeG;omeU;rA;amiA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

730
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7870.18
731

omeC;omeG;omeU;rA;fluA;amiA;omeUs;fluG;omeAs;rGs;fluCSup-3′

731
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7858.15
732

omeC;omeG;omeU;rA;fluA;fluA;amiUs;fluG;omeAs;rGs;fluCSup-3′

732
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7858.15
733

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;amiAs;rGs;fluCSup-3′

733
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7872.17
734

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;amiGs;fluCSup-3′

734
5′-HO;unaG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7875.17
735

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

735
5′-HO;rG;unaC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7875.17
736

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

736
5′-HO;rG;rC;unaUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7875.17
737

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

737
5′-HO;rG;rC;rUs;unaC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7873.18
738

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

738
5′-HO;rG;rC;rUs;fluC;unaA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7861.15
739

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

739
5′-HO;rG;rC;rUs;fluC;omeA;unaU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7873.18
740

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

740
5′-HO;rG;rC;rUs;fluC;omeA;fluU;unaU;rUs;fluA;omeC;omeC;rAs;omeG;
7875.17
741

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

741
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;unaUs;fluA;omeC;omeC;rAs;omeG;
7875.17
742

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

742
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;unaA;omeC;omeC;rAs;omeG;
7873.18
743

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

743
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;unaC;omeC;rAs;omeG;
7861.15
744

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

744
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;unaC;rAs;omeG;
7861.15
745

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

745
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;unaG;
7861.15
746

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

746
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7861.15
747

omeC;omeG;unaU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

747
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7875.17
748

omeC;omeG;omeU;unaA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

748
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7873.18
749

omeC;omeG;omeU;rA;unaA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

749
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7873.18
750

omeC;omeG;omeU;rA;fluA;unaA;omeUs;fluG;omeAs;rGs;fluCSup-3′

750
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7861.15
751

omeC;omeG;omeU;rA;fluA;fluA;unaUs;fluG;omeAs;rGs;fluCSup-3′

751
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7873.18
752

omeC;omeG;omeU;rA;fluA;fluA;omeUs;unaG;omeAs;rGs;fluCSup-3′

752
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7861.15
753

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;unaAs;rGs;fluCSup-3′

753
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7875.17
754

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;unaGs;fluCSup-3′

754
5′-HO;4alk-
7805.03
755

dG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;

omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

755
5′-HO;rG;4alk-
7791.00
756

dC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA;

fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

756
5′-HO;rG;omeC;4alk-
7805.03
757

dU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA;

fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

757
5′-HO;rG;omeC;rU;4alk-
7805.03
758

dC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA;fluA;

omeA;rUs;rG;rAs;rGs;omeCSup-3′

758
5′-HO;rG;omeC;rU;rC;4alk-
7805.03
759

dA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA;fluA;omeA;

rUs;rG;rAs;rGs;omeCSup-3′

759
5′-HO;rG;omeC;rU;rC;rA;4alk-
7803.04
760

dU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA;fluA;omeA;

rUs;rG;rAs;rGs;omeCSup-3′

760
5′-HO;rG;omeC;rU;rC;rA;fluU;4alk-
7805.03
761

dU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA;fluA;omeA;rUs;

rG;rAs;rGs;omeCSup-3′

761
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;4alk-
7805.02
762

dUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;

rAs;rGs;omeCSup-3′

762
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;4alk-
7805.03
763

dA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;

rGs;omeCSup-3′

763
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;4alk-
7803.04
764

dC;rC;rAs;fluG;omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;

omeCSup-3′

764
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;4alk-
7805.02
765

dAs;fluG;omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

765
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;4alk-
7791.00
766

dC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

766
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7805.03
767

4alk-dG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

767
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7803.04
768

rG;4alk-dU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

768
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7791.00
769

fluU;4alk-dA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

769
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7803.04
770

fluU;omeA;4alk-dA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

770
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7791.00
771

fluU;omeA;fluA;4alk-dA;rUs;rG;rAs;rGs;omeCSup-3′

771
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7805.02
772

fluU;omeA;fluA;omeA;4alk-dUs;rG;rAs;rGs;omeCSup-3′

772
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7805.02
773

fluU;omeA;fluA;omeA;rUs;rG;4alk-dAs;rGs;omeCSup-3′

773
5′-HO;ome4SG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7827.09
774

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

774
5′-HO;rG;ome4SC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7813.07
775

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

775
5′-HO;rG;omeC;ome4SU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7827.09
776

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

776
5′-HO;rG;omeC;rU;ome4SC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7827.09
777

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

777
5′-HO;rG;omeC;rU;rC;ome4SA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7827.09
778

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

778
5′-HO;rG;omeC;rU;rC;rA;ome4SU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7825.10
779

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

779
5′-HO;rG;omeC;rU;rC;rA;fluU;ome4SU;rUs;rA;fluC;rC;rAs;fluG;omeC;
7827.09
780

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

780
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;ome4SUs;rA;fluC;rC;rAs;fluG;omeC;
7827.09
781

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

781
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;ome4SC;rC;rAs;fluG;omeC;
7825.10
782

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

782
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;ome4SG;omeC;
7825.10
783

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

783
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;ome4SC;
7813.07
784

rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

784
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;ome4SG;
7827.09
785

fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

785
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7825.10
786

ome4SU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

786
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7813.07
787

fluU;ome4SA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

787
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7825.10
788

fluU;omeA;ome4SA;omeA;rUs;rG;rAs;rGs;omeCSup-3′

788
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7813.07
789

fluU;omeA;fluA;ome4SA;rUs;rG;rAs;rGs;omeCSup-3′

789
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7827.09
790

fluU;omeA;fluA;omeA;ome4SUs;rG;rAs;rGs;omeCSup-3′

790
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7827.09
791

fluU;omeA;fluA;omeA;rUs;ome4SG;rAs;rGs;omeCSup-3′

791
5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;
7827.09
792

fluU;omeA;fluA;omeA;rUs;rG;rAs;ome4SGs;omeCSup-3′

792
5′-HO;4alk-
7881.18
793

dG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;

omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

793
5′-HO;rG;4alk-
7881.18
794

dC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;

omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

794
5′-HO;rG;rC;4alk-
7881.18
795

dUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;

omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

795
5′-HO;rG;rC;rUs;4alk-
7879.19
844

dC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;omeG;

omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

796
5′-HO;rG;rC;rUs;fluC;4alk-
7867.16
796

dA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;omeG;omeU;

rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

797
5′-HO;rG;rC;rUs;fluC;omeA;4alk-
7879.19
797

dU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;omeG;omeU;rA;fluA;

fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

798
5′-HO;rG;rC;rUs;fluC;omeA;fluU;4alk-
7881.18
798

dU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;omeG;omeU;rA;fluA;

fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

799
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;4alk-
7881.18
799

dUs;fluA;omeC;omeC;rAs;omeG;omeC;omeG;omeU;rA;fluA;fluA;

omeUs;fluG;omeAs;rGs;fluCSup-3′

800
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;4alk-
7879.19
800

dA;omeC;omeC;rAs;omeG;omeC;omeG;omeU;rA;fluA;fluA;ome

Us;fluG;omeAs;rGs;fluCSup-3′

801
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;4alk-
7881.18
801

dAs;omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;

rGs;fluCSup-3′

802
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7867.16
802

omeC;4alk-dG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

803
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7867.15
803

omeC;omeG;4alk-dU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

804
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7881.18
804

omeC;omeG;omeU;4alk-dA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

805
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7879.19
805

omeC;omeG;omeU;rA;4alk-dA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

806
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7879.19
806

omeC;omeG;omeU;rA;fluA;4alk-dA;omeUs;fluG;omeAs;rGs;fluCSup-3′

807
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7867.15
807

omeC;omeG;omeU;rA;fluA;fluA;4alk-dUs;fluG;omeAs;rGs;fluCSup-3′

808
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7879.19
808

omeC;omeG;omeU;rA;fluA;fluA;omeUs;4alk-dG;omeAs;rGs;fluCSup-3′

809
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7867.15
809

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;4alk-dAs;rGs;fluCSup-3′

810
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7881.18
810

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;4alk-dGs;fluCSup-3′

811
5′-HO;ome4SG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7903.25
811

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

812
5′-HO;rG;ome4SC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7903.25
812

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

813
5′-HO;rG;rC;ome4SUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7903.24
813

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

814
5′-HO;rG;rC;rUs;ome4SC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;
7901.26
814

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

815
5′-HO;rG;rC;rUs;fluC;omeA;ome4SU;rU;rUs;fluA;omeC;omeC;rAs;
7901.25
815

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

816
5′-HO;rG;rC;rUs;fluC;omeA;fluU;ome4SU;rUs;fluA;omeC;omeC;rAs;
7903.25
816

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

817
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;ome4SUs;fluA;omeC;omeC;rAs;
7903.24
817

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

818
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;ome4SC;omeC;rAs;omeG;
7889.22
818

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

819
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;ome4SAs;
7903.24
819

omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

820
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;ome4SG;
7889.22
820

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

821
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7889.22
821

ome4SC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

822
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7889.22
822

omeC;ome4SG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

823
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7889.22
823

omeC;omeG;ome4SU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

824
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7903.25
824

omeC;omeG;omeU;ome4SA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

825
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7901.26
825

omeC;omeG;omeU;rA;ome4SA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′

826
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7889.21
826

omeC;omeG;omeU;rA;fluA;fluA;ome4SUs;fluG;omeAs;rGs;fluCSup-3′

827
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7901.26
827

omeC;omeG;omeU;rA;fluA;fluA;omeUs;ome4SG;omeAs;rGs;fluCSup-3′

828
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7889.22
828

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;ome4SAs;rGs;fluCSup-3′

829
5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;
7903.24
829

omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;ome4SGs;fluCSup-3′

Example 830

5,-ppp;rG;rC;rU;rC;rA;rU;rU;rU;rA;rC;rC;rA;rG;rC;rG;rU;rA;rA;rA;rU;rG;rA;rG;rCSu p-3′ (SEQ ID No. 839)

To fully protected CPG-bound 5′-O-dimethoxytrityl-oligoribonucleotide (44 μmol), 3% dichloroacetic acid (DCA) in methylene chloride (20 mL) was added and aged at room temperature. The DCA solution was removed after 30 min and the solid support rinsed with acetonitrile (20 mL). After removal of the acetonitrile, solutions of 5-benzylthio-1H-tetrazole (3504 μL, 0.876 μmol) and 6-chloro-N,N-diisopropyl-4H-benzo[d][1,3,2]dioxaphosphinin-2-amine (4380 μL, 0.876 μmol) in acetonitrile were added and the reaction was aged at room temperature overnight. The solution was removed from the solid support and 0.02 M I₂in THF/H₂O/pyridine (Glen Research, cat: 40-4330-52) was added and the reaction was aged for 5 min. The oxidizer reagent was removed and the solid support was rinsed with acetonitrile (20 mL). After removal of acetonitrile, the CPG was dried under vacuum for 4 h. To the solid support was added 0.5 M tetrabutylammonium dihydrogendiphosphate in dry DMF (3.5 mL) and the mixture was aged at 45° C. for 3 days. The solid was filtered and washed with DMF, water (3×), and acetonitrile. The solid support then was treated with 8 mL AMA (40% aqueous methylamine:concentrated aqueous ammonia hydroxide, 1:1 v/v) for 2.5 h. The CPG was filtered and washed with an additional 5 mL AMA. The combined filtrates were placed in a GeneVac (ambient temperature) overnight to remove all solvent. The resulting pellet was treated with 5 mL dry THF and TBAF (tetra-n-butylammonium fluoride) in dry THF (8 mL, 1.0 M), and the mixture was aged at room temperature overnight. The reaction was quenched with 2 mL concentrated aqueous ammonia hydroxide. The solvent was removed from the reaction with a GeneVac evaporator (ambient temperature) for 5 h. The resulting viscous material was diluted with 40 mL water and dialyzed by spin filtration (3000 MWCO). The solution was further washed by spin dialysis with water (4×40 mL). The retained solution was collected, diluted with water to 15 mL and purified by ion pairing reversed phase (IP-RP) HPLC using a YMC column, 19×150 mm, 5 μm, to give the desired product.

HR LC-MS: (m/z)⁻(z=4) 1969.2855 (exp); 1969.2442 (theor).

Examples 830-837 as shown in Table 6 below were prepared according to the procedures analogous to those outlined in Example 830 above

TABLE 6

SEQ

ID

Ex#
Sequence
MW
No.

830
ppp;rG;rC;rU;rC;rA;
7876.52
1

rU;rU;rU;rA;rC;rC;rA;

rG;rC;rG;rU;rA;rA;rA;

rU;rG;rA;rG;rCSup

831
ppp;rG;omeC,rU;rC;rA;
7956.61
1

fluU;rU;rU;rA;fluC;rC;

rA;fluG;omeC;rG;fluU;

omeA,fluA;omeA;rU;rG;

rA;rG;omeCSup

832
ppp;rG;omeCs;rUs;rCs;
8085.13
1

rA;fluU;rU;rUs;rA;

fluC;rC;rAs;fluGs;

omeCs;rG;fluU;omeA;

fluAs;omeA;rU;rG;rA;

rG;omeCSup

833
ppp,rG;omeC;rU;rC;rA;
8036.94
1

fluU;rU;rUs;rA;fluC;

rC,rAs;fluG,omeC;rG;

fluU;omeA;fluA;omeA;

rUs;rG;rAs;rGs;omeCSup

834
ppp;rGs;rCs;omeUs;rCs;
8177.45
1

fluAs;rUs;omeUs;rUs;

rAs;rCs;omeC;rA;fluG;

omeC;rG;omeU;omeA;

fluA;fluA;rU;rGs;

omeA,rGs;fluCSup

835
ppp;rG;rC;rU;fluC,
8097.03
1

omeA;fluU;rU;rUs;fluA;

omeG;omeC;rAs;omeG;

omeC,omeG;omeU;rA;

fluA;fluA;omeUs;fluG,

omeAs;rGs;fluCSup

836
ppp,rG;rC;rU;fluC;
8016.70
1

omeA;fluU;rU;rU;fluA;

omeC;omeC;rA;omeG;

omeC;omeG;omeU;rA;

fluA;fluA;omeU;fluG;

omeA;rG;fluCSup

837
ppp;rG;rC;rUs;fluC;
8113.09
1

omeA;fluU;rU;rUs;fluA;

omeC;omeC;rAs;omeG;

OmeC;omeG;omeU;rA;

fluA,fluA;omeUs;fluG;

omeAs;rGs;fluCSup

Example 838

5′-PcP;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;omeG;omeU;r A;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ (SEQ ID No. 840)

Fully protected CPG-bound 5′-O-dimethoxytrityl-oligoribonucleotide (5′-DMT-O;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;omeG;omeU;rA;fluA;f luA;omeUs;fluG;omeAs;rGs;fluCSup-3′ (SEQ ID NO: 842); 10 μmol) was treated with 3% 2′,2′-dichloroacetic acid in dichloromethane for 5 min (2×5 mL). Then, the solid phase was washed with acetonitrile (3×3 mL) and dichloromethane (3×3 mL). To the solid phase, N₂was passed through for 2 min and it was kept under vacuum for 1 h.

The solid phase was treated with diethyl (((diisopropylamino)(methoxy)phosphaneyl)methyl)phosphonate (0.125 g, 0.400 mmol), acetonitrile (2.5 mL), and 5-(ethylthio)-2H-tetrazole (20 mg, 0.15 mmol) in acetonitrile (0.5 mL). The mixture was shaken at room temperature for 18 h. Then, the solid phase was filtered, rinsed with acetonitrile (2×1 mL), and treated with 12 in acetonitrile/pyridine/water (0.025 M, 9/9/2, 3 mL) for 5 min. The resin was further rinsed with acetonitrile (4×3 mL) and dichloromethane (2×3 mL). The solid phase was dried by passing a stream of N₂through it for 1 min and kept under vacuum for 1 h.

The resulting solid phase was treated with TMSI (20 mg, 0.10 mmol) in pyridine/acetonitrile (v/v 1/50, 3 mL) for 30 min. Then, the solid phase was filtered, rinsed with acetonitrile (2×3 mL) and treated with 2-mercaptoethanol in Et₃N/acetonitrile (1 M, v/v 1/1, 0.5 mL) for 1 min. The CPG was washed again with acetonitrile (5×5 mL) and then treated with 40% aq MeNH₂(2 mL) for 20 min. The solid support was filtered off and rinsed with 40% aq MeNH₂(2×2 mL). The combined filtrate was shaken at ambient temperature for 2 h and then partly concentrated to ca. 2 mL under reduced pressure. The resulting solution lyophilized to give a solid.

The solid was dissolved in water (4 mL) and Et₃N-3HF (0.32 mL, 2.0 mmol) in water (1 mL) was added. The mixture was shaken at 40° C. for 6 h, after which aqueous sodium acetate (6 mL, 1 M) was added. The resulting mixture was purified by preparative HPLC (C18), eluting with 5 to 40% acetonitrile in water (0.1 M triethylammonium acetate) to provide the desired product.

LC-MS: (m/z)⁻(z=5) 1605.7 (exp), 1605.2 (theor); (z=6) 1338.0 (exp), 1337.5 (theor); (z=7) 1146.3 (exp), 1146.3 (theor); (z=8) 1002.9 (exp), 1002.9 (theor).

The oligonucleotides of the invention based on SEQ ID No. 1 comprises at least one 5′ group selected from L in Table 2a; 24 unmodified nucleobases consisting of (Adenine (A), Guanine (G), Cytosine (C) and Uracil (U), and when read from left to right, is presented as 5′-GCUCAUUUACCAGCGUAAAUGAGC-3′ (SEQ ID NO: 841) without the L group. Certain oligonucleotides of the invention based on SEQ ID No. 1 further comprise optionally modified sugar groups and optionally modified phosphodiester groups. One or more of the sugar groups optionally comprises modifications at one or more of the 2′ and/or 4′ positions selected from Table 2 for the first position nucleotide, Tables 3 and 3a for positions 2-23 nucleotides, and Table 4 for position 24 nucleotide. Exemplary modifications at the 2′ position are selected from the group consisting of OH, CH₃, OCH₃, OCF₃, OP(O)OH, O(CH₂)₂OCH₃, fluoro, NH₂, locked nucleosides, or 2′-3′ seco-nucleosides. Exemplary modifications of the 4′ position are selected from halogen, C_1-6alkyl, or C_2-6alkynyl. The oligonucleotides of the invention may also comprise modified phosphodiester backbone groups also depicted in Tables 3 and 3a.

An embodiment of the invention of Formula Ia and Formula I is realized when greater than 10 independently selected modified nucleobases from Table 3 are incorporated into the nucleotide sequence. A subembodiment of this aspect of the invention is realized when 10 to 20, 10 to 15, or 10 to 12 independently selected modified nucleobases from Table 3 are incorporated into the nucleotide sequence. Another subembodiment of this aspect of the invention is realized when the nucleotide sequence comprises optionally modified sugar groups and optionally modified phosphodiester backbones. A further aspect of this subembodiment of the invention is realized when one or more of the sugar groups optionally comprises modifications at one or more of the 2′ and/or 4′ positions selected from Table 2 for the first position nucleotide, Table 3 for positions 2-23 nucleotides, and Table 4 for position 24 nucleotide. Still a further aspect of this subembodiment of the invention is realized when modifications at the 2′ position are selected from the group consisting of OH, CH₃, OCH₃, OCF₃, OP(O)OH, O(CH₂)₂OCH₃, fluoro, NH₂, locked nucleosides, 2′-3′ seco-nucleosides and modifications of the 4′ position are selected from halogen, C_1-6alkyl, or C_2-6alkynyl.

An embodiment of the invention of Formula Ia and Formula I is realized when one or more of the sugar moieties at nucleotide positions 1, 2, 3, 7, 8, 12, and/or 17 optionally comprise a natural nucleobase; one or more of the sugar moieties at nucleotide positions 4, 6, 9, 18, 19, 21 and/or 24 optionally comprise a modified nucleobase containing a 2′ fluoro group; one or more of the sugar moieties at nucleotide positions 5, 10, 11, 13, 14, 15,16, 20, and/or 22 optionally comprise a modified nucleobase containing 2′ OMe group; and Formula Ia and Formula I optionally comprises a modified phosphodiester backbone between positions 3-4, 8-9, 20-21, 22-23 and/or 23-24.

Another embodiment of the invention of Formula Ia and Formula I is realized when SEQ ID No. 1 optionally comprises an 2′-OH at one or more of the sugar moieties at nucleotide positions 1, 3, 4, 5, 7, 8, 9, 11, 12, 15, 20, 22, and/or 23; optionally comprises fluoro at one or more of the sugar moieties at nucleotide positions 6, 10, 13, 16, and/or 18; optionally comprises 2′-OMe at one or more of the sugar moieties at nucleotide positions 2, 14, 17, 19, and/or 24; and optionally comprises a modified phosphodiester backbone between positions 8-9, 12-13, 20-21, 22-23 and/or 23-24.

An embodiment of the invention of Formula Ia and Formula I is realized when the position 8 nucleotide is selected from the group consisting of rUs, thiorUs, araU, and d5propUs.

Another embodiment of the invention of Formula Ia and Formula I is realized when the position 12 nucleotide is selected from rAs, d7dzAs, cf3As, and d2aminoAs.

Another embodiment of the invention of Formula Ia and Formula I is realized when the position 2 nucleotide is rC.

Another embodiment of the invention of Formula Ia and Formula I is realized when the position 3 nucleotide is selected from the group consisting of rUs and thiorUs.

Another embodiment of the invention of Formula Ia and Formula I is realized when the position 7 nucleotide is selected from the group consisting of rU and thiorU.

Another embodiment of the invention of Formula Ia and Formula I is realized when the position 22 nucleotide is selected from the group consisting of omeAs and araAs.

Still another embodiment of the invention of Formula Ia and Formula I is realized when the terminal 5′ substituent is selected from OH and triphosphate. A subembodiment of this aspect of the invention is realized when the terminal 5′ substituent is OH. Another subembodiment of this aspect of the invention is realized when the terminal 5′ substituent is triphosphate.

Yet another embodiment of the invention of Formula Ia and Formula I is realized when the terminal 5′ end is OH or triphosphate, nucleotide positions 1, 4, 5, 6, 9, 10, 11, 13, 14, 15, 16, 17, 18, 19, 20, 21, 23 and 24, respectively, is:

rG;rC;rUs;fluC;omeA;fluU;rU;ome4SUs;fluA;omeC;

omeC;rAs;omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup (SEQ ID NO: 843). A subembodiment of this aspect of the invention is realized when nucleotide position 8 is selected from the group consisting of rUs, thiorUs, araUs, and d5propUs. Another subembodiment of this aspect of the invention is realized when nucleotide position 12 is selected from the group consisting of rAs, d7dzAs, cf3As, and d2aminoAs. Another subembodiment of this aspect of the invention is realized when nucleotide position 2 is selected from the group consisting of rC. Another subembodiment of this aspect of the invention is realized when nucleotide position 3 is selected from the group consisting of rUs and thiorUs. Another subembodiment of this aspect of the invention is realized when nucleotide position 7 is selected from the group consisting of rU and thiorU.

Biological Data

The oligonucleotides described here are RIG-I agonists which demonstrate interferon production with an EC₅₀of <1.7 μM. Preferred oligonucleotides of the invention demonstrate EC50 values of <1.0 μM, <0.5 μM, <0.25 μM, <0.1 μM, or <0.05 μM in the A549 Dual cell assay when transfected using LyoVec. The methods below describe each of these assays.

Annealing Hairpin RNA

All hairpin RNA samples were annealed and snap cooled prior to testing. RNA samples (100 uM) were prepared in phosphate buffered saline (PBS) and transferred to a PCR plate. The RNA containing plate was sealed, placed in a thermocycler and heated at 95° C. for 1 minute followed by immediate transfer to wet ice. The plate was incubated on ice for 10 minutes and then allowed to return to room temperature prior to subsequent reformatting.

A549 Dual Cell Assay

Equilibrate a 96-well source plate(s) containing 10 nmol of lyophilized RNA in each well to room temperature. Centrifuge the plate(s) at 800×g for 5 minutes at room temperature to pellet material to the bottom of the wells. Carefully remove plate seal(s) and resuspend the RNA using 100 μL of molecular biology grade 1×PBS, hereafter referred to as 1×PBS (Thermo Fisher; Cat. No. AM9624). Mix thoroughly to ensure complete solvation of RNA. Allow the RNA plate(s) to stand for 5-10 minutes at room temperature to aid complete dissolution of all RNA. At this point the final concentration of RNA after dissolution is 100 μM in 100 μL.

To prepare dilution series of the RNA, briefly mix then transfer 60 μL of 10 μM positive control RNA (ppp-SLR10, Pyle et al. Sci. Adv. 2018 February; 4 (2): e1701854) previously prepared in 1×PBS, or test RNA solution from the 96-well source plate(s) to the appropriate wells in columns 3 and 13 of 384-well DNA Lo-Bind destination plate(s) (Eppendorf, Cat. No. 951040546). To columns 4-12 and 14-22 of the destination plate(s) add 30 μL of 1×PBS. Transfer 15 μL of RNA solution from columns 3 and 13 to adjacent wells in columns 4 and 14 and mix by aspiration. Using fresh pipette tips each time, repeat this process a further 8 times until columns 12 and 22 are reached to generate 10-point, 3-fold serial dilutions. In wells A1 to H2 and 123 to P24 add 45 μL 10 μM control RNA to define maximum effect. In wells I1 to P2 and I23 to P24 add 45 μL 1×PBS to define minimum effect. At this point these serial dilution plates can be stored at −20° C. until required.

To further dilute the RNA samples transfer 3 μL of RNA solution from all wells of the serial dilution plate(s) to fresh 384-well DNA LoBind plate(s). To these new plate(s) add 27 μL of molecular biology grade 1×PBS and mix. These plates can be stored at −20° C. until required. These 1:10 dilution daughter plates are used as the RNA source plates in this assay and can be sealed and kept at room temperature if being used on the same day or sealed and stored at −20° C. until needed.

Transfer 5 μL from each well of the 1:10 dilution daughter plate(s) to equivalent wells of 384-well assay plate(s) (Corning; Cat. No. 3764). The plates are sealed and kept at room temperature for up to 2 hours. Dissolve LyoVec transfection reagent (Invivogen; Cat. No. lyec-2) to 0.078 mg/mL using 1×PBS. Dispense 5 μL of LyoVec solution into each well of 384-well assay plate(s). Briefly centrifuge the plate(s) to ensure the RNA/LyoVec mixture is brought down to the bottom of the wells and incubate for a minimum of 15 minutes before adding cells. Rapidly thaw A549-Dual cells (originally obtained from Invivogen; Cat. No. a549d-nfisi), dilute in 5 volumes of prewarmed assay buffer (DMEM, Thermo Fisher Cat. No. 21063-029; supplemented with 10% FBS, Thermo Fisher, Cat. No. 10091148), perform a cell count on a small sample of the cell suspension and centrifuge the remainder at 200×g for 5 minutes at room temperature. Resuspend the cell pellet to a density of 2 million viable cells per mL in assay buffer. Add 20 μL of cells per well to all wells of the 384-well assay plate(s). Briefly, centrifuge the plates to ensure that the cell suspension is brought down to the bottom of the wells. Incubate the plates at 37° C., 5% CO₂in a humidified chamber for 18 hours.

Protecting the reagent from light, reconstitute the Quanti-Luc detection reagent (Invivogen; Cat. No. rep-qlc2) with 25 mL of deionized water per reagent sachet. Remove assay plate(s) from the incubator and transfer 10 μL of supernatant from the assay plate(s) to 384-well AlphaPlate(s) (Perkin Elmer; Cat. No. 6005359). Then add 25 μL of Quanti-Luc reagent to each well of the AlphaPlate(s) and mix thoroughly. Incubate each AlphaPlate for 60 minutes at room temperature then read plate on an Envision Multilabel Plate Reader (Perkin Elmer) using a 384-L1 aperture, an integration time of 0.2 seconds and reading by rows bi-directionally.

Data is normalized to the maximum and minimum effect control wells and then subjected to a 4 parameter logistic fit based on the Levenberg-Marquardt algorithm to provide measures of potency and relative efficacy.

Examples 1 through 838 were evaluated in cell based assay as described above. Table 7 gives the biological data (RIG-I EC₅₀) collected for each example using the methodology described above.

Compounds of the disclosure were evaluated for interferon β production in A549 cell culture with LyoVec as described above. The following Table 7 tabulates the biological data for these compounds as EC₅₀.

TABLE 7

Ex
EC₅₀(μM)

1
0.002

2
0.007

3
0.012

4
0.104

5
0.197

6
0.003

7
0.069

8
0.007

9
0.006

10
0.139

11
0.007

12
0.037

13
0.029

14
0.010

15
0.049

16
0.039

17
0.016

18
0.014

19
0.061

20
0.099

21
0.009

22
0.066

23
0.176

24
0.214

15
0.058

26
0.148

27
0.016

28
0.016

29
0.018

30
0.023

31
0.005

32
0.273

33
0.005

34
0.481

35
0.029

36
0.247

37
0.048

38
0.018

39
0.008

40
0.118

41
0.169

42
0.011

43
0.172

44
0.219

45
0.038

46
0.239

47
0.066

48
0.024

49
0.101

50
0.195

51
0.080

51
0.056

53
0.350

54
0.075

55
0.022

56
0.219

57
0.011

58
0.039

59
0.375

60
0.074

61
0.121

62
0.012

63
0.008

64
0.231

65
0.006

66
0.028

67
0.036

68
0.295

69
0.084

70
0.075

71
0.422

72
0.243

73
0.250

74
0.339

75
0.194

76
0.158

77
0.038

78
0.392

79
0.121

80
0.381

81
0.310

82
0.092

83
0.280

84
0.083

85
0.011

86
0.284

87
0.012

88
0.211

89
0.295

90
0.080

91
1.122

92
0.014

93
0.011

94
0.013

95
0.013

96
0.008

97
0.012

98
0.109

99
0.018

100
0.011

101
0.014

102
0.017

103
0.128

104
1.177

105
0.104

106
0.235

107
0.240

108
0.014

109
0.021

110
0.084

111
0.014

112
0.021

113
0.073

114
0.469

115
0.292

116
0.054

117
0.137

118
0.005

119
0.026

120
0.010

121
0.029

122
0.112

123
0.139

124
0.235

125
0.322

126
1.091

127
0.253

128
0.291

129
0.077

130
0.042

131
0.120

132
0.038

133
0.368

134
0.070

135
0.114

136
1.238

137
0.160

138
0.070

139
0.065

140
0.092

141
0.319

142
0.027

143
0.272

144
0.374

145
0.055

146
0.144

147
0.121

148
0.422

149
0.013

150
0.029

151
0.551

152
0.042

153
0.022

154
0.262

155
0.377

156
0.167

157
0.200

158
0.465

159
0.065

160
0.157

161
0.143

162
0.043

163
0.384

164
0.442

165
0.246

166
0.017

167
0.442

168
0.387

169
0.076

170
0.028

171
0.538

172
0.361

173
0.206

174
0.463

175
1.101

176
0.592

177
0.502

178
0.796

179
0.002

180
0.002.

181
0.002

182
0.001

183
0.001

184
0.001

185
0.002

186
0.001

187
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188
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189
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190
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191
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192
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193
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194
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195
0.001

196
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197
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198
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199
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200
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201
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202
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203
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204
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205
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206
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207
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208
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209
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210
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211
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212
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213
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216
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217
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220
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224
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225
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226
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227
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228
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229
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230
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231
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232
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233
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234
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235
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236
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237
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238
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239
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240
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241
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242
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244
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246
0.001

247
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248
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249
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250
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251
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252
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253
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255
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256
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257
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258
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259
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260
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261
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262
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263
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264
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265
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266
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267
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268
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269
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270
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271
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/72
0.002

273
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274
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275
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276
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277
0.004

278
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279
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280
0.003

281
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282
0.013

283
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284
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285
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286
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287
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288
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289
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290
0.003

291
0.005

292
0.002

293
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294
0.006

295
0.002

296
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297
0.004

298
0.003

299
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300
0.002

301
0.002

302
0.002

303
0.002

304
0.002

305
0.002

306
0.006

307
0.002

308
0.023

309
0.051

310
0.004

311
0.001

312
0.003

313
0.008

314
0.004

315
0.004

316
0.864

317
0.007

318
0.040

319
0.006

320
0.016

321
0.007

322
0.010

323
0.003

324
0.009

325
0.001

326
0.005

327
0.004

328
0.012

329
0.018

330
0.015

331
0.095

332
0.009

333
0.005

334
0.003

335
<0.001

336
0.008

337
0.004

338
0.003

339
0.005

340
0.003

341
0.003

342
0.005

343
0.019

344
<0.001

345
0.003

346
0.001

347
0.006

348
0.002

349
0.002

350
0.001

351
0.002

352
0.015

353
0.006

354
0.004

355
0.004

356
0.005

357
0.003

358
0.016

359
0.004

360
0.008

361
0.028

362
0.003

363
0.003

364
0.006

365
0.037

366
0.021

367
0.055

368
0.005

369
0.012

370
0.004

371
0.001

372
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373
0.003

374
0.002

375
0.002

376
0.004

377
0.002

378
0.001

379
0.001

380
0.011

381
0.002

382
0.004

383
<0.001

384
0.005

385
0.007

386
0.005

387
0.001

388
0.005

389
0.013

390
0.020

391
0.011

392
0.015

393
0.010

394
0.003

395
0.012

396
0.005

397
0.005

398
0.010

399
0.008

400
0.006

401
0.011

402
0.007

403
0.006

404
0.011

405
0.004

406
0.005

407
0.008

408
0.004

409
0.003

410
0.005

411
0.005

412
0.006

413
0.004

414
0.005

415
0.004

416
0.005

417
0.005

418
0.004

419
0.004

420
0.003

421
0.004

422
0.006

423
0.002

424
0.006

425
0.007

426
0.010

417
0.008

428
0.004

429
0.009

430
0.005

431
0.009

432
0.003

433
0.002

434
0.004

435
0.004

436
0.010

437
0.006

438
0.006

439
0.005

440
0.008

441
0.011

442
0.007

443
0.010

444
0.012

445
0.006

446
0.004

447
0.004

448
0.001

449
0.004

450
0.003

451
0.002

452
0.002

453
0.007

454
0.029

455
0.004

456
0.003

457
0.005

458
0.004

459
0.003

460
0.004

461
0.003

462
0.004

463
0.002.

464
0.005

465
0.005

466
0.009

467
0.004

468
0.003

469
0.006

470
0.007

471
0.011

472
0.011

473
0.005

474
0.011

475
<0.001

476
0.002

477
0.004

478
0.002

479
0.008

480
0.004

481
0.004

482
0.004

483
0.004

484
0.004

485
0.004

486
0.003

487
0.003

488
0.004

489
0.003

490
0.002

491
0.004

492
0.003

493
0.002

494
0.003

495
0.002

496
0.003

497
<0.001

498
0.002

499
0.004

500
0.003

501
0.003

502
0.004

503
0.003

504
0.003

505
0.003

506
0.008

507
0.003

508
0.003

509
0.003

510
0.004

511
0.004

512
0.003

513
0.017

514
<0.001

515
0.003

516
0.002

517
0.003

518
0.001

519
0.005

520
0.010

521
0.009

522
0.026

523
0.033

524
1.111

525
1.667

526
0.009

527
0.008

528
0.008

529
0.004

530
0.005

531
0.005

532
0.004

533
0.004

534
0.003

535
0.004

536
0.002

537
0.004

538
0.022

539
0.004

540
0.004

541
0.005

542
0.004

543
0.003

544
0.004

545
0.005

546
0.004

547
0.004

548
0.004

549
0.008

550
0.005

551
0.004

552
0.011

553
0.004

554
0.004

555
0.008

556
0.382

557
0.007

558
0.010

559
0.004

560
0.004

561
0.004

562
0.004

563
0.018

564
0.013

565
0.006

566
0.007

567
0.004

568
0.006

569
0.004

570
0.011

571
0.009

572
0.004

573
0.034

574
0.008

575
0.012

576
0.004

577
0.009

578
0.012

579
0.007

580
0.010

581
0.010

582
0.007

583
0.005

584
0.005

585
0.004

586
0.008

587
0.004

588
0.012

589
0.009

590
0.009

591
0.010

592
0.013

593
0.005

594
0.014

595
0.011

596
0.012

597
0.037

598
0.040

599
0.037

600
0.029

601
0.037

602
0.018

603
<0.001

604
0.007

605
0.006

606
0.006

607
0.004

608
0.006

609
0.029

610
0.028

611
0.027

612
0.032

613
0.037

614
0.011

615
0.005

616
0.005

617
0.049

618
0.043

619
0.003

620
0.004

621
0.003

622
0.011

623
0.010

624
0.009

625
0.013

626
0.054

627
0.025

628
0.036

629
0.038

630
0.002

631
<0.001

632
0.001

633
0.001

634
0.006

635
0.011

636
0.012

637
0.036

638
0.031

639
0.032

640
0.024

641
<0.001

642
0.002

643
0.001

644
0.009

645
0.004

646
0.004

647
0.006

648
0.005

649
0.023

650
0.019

651
0.026

652
0.033

653
0.002

654
0.003

655
<0.001

656
0.002

657
0.040

658
0.042

659
0.006

660
0.002

661
0.009

662
0.007

663
0.011

664
0.011

665
0.036

666
0.037

667
0.033

668
0.038

669
<0.001

670
0.002

671
0.002

672
0.007

673
0.015

674
0.007

675
0.004

676
0.003

677
0.006

678
0.003

679
0.008

680
0.012

681
0.004

682
0.003

683
0.005

684
0.006

685
0.006

686
0.003

687
0.012

688
0.005

689
0.006

690
0.009

691
0.167

692
0.009

693
0.017

694
0.005

695
0.009

696
0.005

697
0.011

698
0.014

699
0.012

700
0.002

701
0.002

702
0.001

703
<0.001

704
0.005

705
0.013

706
0.011

707
0.006

708
0.006

709
0.008

710
0.007

711
0.010

712
0.014

713
0.011

714
0.003

715
0.003

716
0.004

717
0.004

718
0.005

719
0.007

720
0.003

721
0.008

722
0.005

723
0.003

724
0.004

725
0.003

726
0.007

727
0.003

728
0.004

729
0.004

730
0.005

731
0.007

732
0.010

733
0.006

734
0.010

735
0.111

736
0.014

737
0.142

738
0.007

739
0.013

740
0.009

741
0.009

742
0.012

743
0.007

744
0.006

745
0.001

746
0.015

747
0.012

748
0.005

749
0.010

750
0.007

751
0.011

752
0.004

753
0.009

754
0.014

755
0.012

756
0.003

757
0.007

758
0.002

759
0.005

760
0.015

761
0.011

761
0.019

763
0.009

764
0.016

765
0.007

766
0.004

767
0.009

768
0.035

769
0.007

770
0.088

771
0.019

772
0.022

773
0.062

774
0.004

775
0.002

776
0.010

777
0.003

778
0.005

779
0.008

780
0.042

781
0.009

782
0.003

783
0.003

784
0.002

785
0.006

786
0.005

787
0.007

788
0.011

789
0.011

790
0.007

791
0.023

792
0.007

793
0.010

794
0.009

795
0.006

796
0.004

797
0.009

798
0.041

799
0.017

800
0.013

801
0.004

802
0.009

803
0.006

804
0.015

805
0.009

806
0.015

807
0.009

808
0.008

809
0.013

810
0.013

811
0.013

812
0.011

813
0.011

814
0.005

815
0.008

816
0.012

817
0.007

818
0.013

819
0.003

820
0.008

821
0.006

822
0.009

823
0.004

824
0.009

825
0.009

826
0.007

827
0.016

828
0.011

829
0.025

830
<0.001

831
<0.001

832
0.001

833
<0.001

834
NA

835
NA

836
0.002

837
0.004

838
0.001

Human Peripheral Blood Mononuclear Cell (PBMC) Assay

The cell-based potencies of representative compounds of the disclosure were evaluated for interferon α production in human PBMC cell culture with LyoVec as described above compared to known literature RIG-I agonist, 5′ppp-GGACGUACGUUUCGACGUACGUCC-3′, referred to as SEQ ID No. 123 herein, and as SEQ ID No. 12 in US2016/0046943 A (5ppp 10 L); also in Linehand et al. Sci Adv. 2018 February; as 4(2) (ppp-SLR10)) were ascertained by measuring the increase of the type I interferon, IFN-α, secreted by human peripheral blood mononuclear cells (PBMCs) following transfection of the test RNAs.

The test RNAs were serially diluted in calcium/magnesium-free 1×DPBS (Gibco Cat. No. 14190136) across 10 points with 5 μL per dilution point added to the wells of a flat-bottom tissue-culture treated 384-well plate (Corning Cat. No. 3764). Five microliters per well of 1:50 diluted transfection reagent Lipofectamine 2000 (Invitrogen Cat. No. 11668019) were added to each RNA-containing. The RNA/Lipofectamine 2000 mixtures were incubated at room temperature for 10 minutes to enable complexation. Frozen human PBMCs (AllCells Cat. No. C-PB102F-001) were thawed, resuspended to a density of 2.5×10⁶/mL in assay medium (RPMI 1640 (Gibco Cat. No. 11875-085) supplemented with 10% fetal bovine serum (Gibco Cat. No. 10091148)) then 40 μL was added to each well containing the complexed RNA.

After 18 hours of incubation at 37° C., 5% CO₂, 5 μL of undiluted cell supernatant was transferred from each well into a 384-well AlphaPlate (Perkin Elmer Cat. No. 6005359). Interferon alpha levels were measured using a human IFN-α AlphaLISA Kit (Perkin Elmer Cat. No. AL297F) following the manufacturer's protocol. Briefly, 10 μL of 5× AlphaLISA anti-IFN-α acceptor beads were added to each supernatant-containing well followed by a 30 minute room temperature incubation. Ten microliters of 5× biotinylated anti-IFN-α antibody were then added to each well followed by a 60 minute room temperature incubation. Twenty-five microliters of light-sensitive 2× Streptavidin donor beads were added to each well followed by a final incubation of 30 minutes at room temperature in the dark. A Perkin Elmer Envision Multilabel Plate Reader with a 384-A1 aperture was used to scan the plates at an excitation wavelength of 680 nM and emission wavelength of 615 nM. The data were analyzed using a nonlinear regression model, log(agonist) vs. response—Variable Slope, in Prism 7.05 (GraphPad Software, La Jolla, Calif., USA). The following Table 8 tabulates the biological data for these compounds as EC₅₀.

Table 8: Biological activities of unmodified example 830 and modified example 833 compared to literature RIG-I agonist 5ppp 10 L (pppSLR10) in human PBMC assay.

TABLE 8

95% CI

EC50
Lower
Upper

(nM)
Limit
Limit

5ppp 10L (pppSLR10)

Donor 1
6.84
5.65
8.22
Mean EC50 (95% CI)

Donor 2
7.79
5.41
10.74
nM = 6.07 (4,72, 7.75)

Donor 3
4.23
3.63
4.98

Donor 4
6.04
4.48
8.21

Example 830

Donor 1
0.41
0.32
0.54
Mean EC50 (95% CI)

Donor 2
0.34
0.25
0.46
nM = 0.36 (0.25, 0.51)

Donor 3
0.37
0.22
0.59

Donor 4
0.32
0.22
0.46

Example 833

Donor 1
0.80
0.61
1.07
Mean EC50 (95% CI)

Donor 2
0.49
0.38
0.65
nM = 0.64 (0.46, 0.93)

Donor 3
0.67
0.48
0.94

Donor 4
0.63
0.39
1.14

Number	Name	Date	Kind
7488802	Collins et al.	Feb 2009	B2
7521051	Collins et al.	Apr 2009	B2
7655785	Bentwich	Feb 2010	B1
8008449	Korman et al.	Aug 2011	B2
8168757	Finnefroch et al.	May 2012	B2
8354509	Carven et al.	Jan 2013	B2
8383796	Korman et al.	Feb 2013	B2
9439962	Honjo et al.	Sep 2016	B2
20080025979	Honjo et al.	Jan 2008	A1
20090111765	Hartmann et al.	Apr 2009	A1
20110271358	Freeman et al.	Nov 2011	A1
20130037977	Burke et al.	Feb 2013	A1
20160152983	Goldeck et al.	Jun 2016	A1

Number	Date	Country
3045536	Jul 2016	EP
2004004771	Jan 2004	WO
2004056875	Jul 2004	WO
2004072286	Aug 2004	WO
2010027827	Mar 2010	WO
2010077634	Jul 2010	WO
2011066342	Jun 2011	WO
2013019906	Feb 2013	WO
2014159990	Oct 2014	WO
2016179034	Nov 2016	WO
2017173427	Oct 2017	WO
2019204743	Oct 2019	WO

Substituted RIG-I agonists: compositions and methods thereof

Information

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Date Filed

Date Issued

Inventors

Original Assignees

Examiners

Agents

CPC

Field of Search

CPC

International Classifications

Term Extension

Abstract

Description

Claims

CROSS-REFERENCE TO RELATED APPLICATIONS

PCT Information

US Referenced Citations (13)

Foreign Referenced Citations (12)

Non-Patent Literature Citations (30)

Provisional Applications (1)

Entry
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