INHBE GENE-TARGETTING DOUBLE-CHAIN ​​OLIGONUCLEOTIDE, ITS CONJUGATE, COMPOSITION AND APPLICATION

Claims

1. A double-stranded oligonucleotide that targets the INHBE gene, wherein the double-stranded oligonucleotide comprises a sense strand and an antisense strand, wherein the antisense strand is complementary or substantially complementary to the sense strand; wherein said "substantially complementary" means that the sense strand and the antisense strand have mismatches of no more than 3 nucleotides in a duplex region; wherein the sense strand comprises a nucleotide sequence that is identical or substantially identical to at least 15 contiguous nucleotides in the sequence of SEQ ID NO: 309; and said "substantially identical" means that there are no more than 3 nucleotide differences between the sense strand and at least 15 contiguous nucleotides in the sequence of SEQ ID NO:

309.

2. The double-stranded oligonucleotide of claim 1, wherein the sense strand comprises a nucleotide sequence having no more than 2 nucleotide differences from at least 15 adjacent nucleotides in the sequence SEQ ID NO: 309, preferably no more than 1 nucleotide difference; optionally, the antisense strand is complementary or substantially complementary to at least 15, 16, 17, 18, 19, 20, 21, 22, 23 contiguous nucleotides of the nucleotide sequence of SEQ ID NO: 309, wherein said "substantially complementary" means having a mismatch of no more than 3 nucleotides in the region of complementarity; optionally, the antisense strand comprises at least 15 contiguous nucleotides of any of the sequences presented in SEQ ID NO: 155 - SEQ ID NO: 308 in Table 1, or a nucleotide sequence having no more than 3 nucleotide differences from at least 15 contiguous nucleotides; wherein the sense strand comprises a nucleotide sequence that is at least partially reverse complementary or substantially complementary to the antisense strand to form a duplex region; wherein the said "substantially complementary" means that the sense strand and the antisense strand have mismatches of no more than 3 nucleotides in the duplex region; optionally, the antisense strand comprises at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21 contiguous nucleotides of any of the nucleotide sequences presented in SEQ ID NO: 155 - SEQ ID NO: 308 in Table 1, or a nucleotide sequence having no more than 3 nucleotide differences from the contiguous nucleotides; wherein the antisense strand optionally comprises at least 17, at least 18, at least 19, at least 20, at least 21 contiguous nucleotides of any of the nucleotide sequences presented in SEQ ID NO: 155 - SEQ ID NO: 308 in Table 1, or a nucleotide sequence having no more than 2 nucleotide differences from the contiguous nucleotides; wherein the antisense strand optionally comprises at least 19, at least 20, at least 21 contiguous nucleotides of any of the nucleotide sequences presented in SEQ ID NO: 155 - SEQ ID NO: 308 in Table 1, or a nucleotide sequence having no more than 1 nucleotide difference from the contiguous nucleotides; wherein the antisense strand is optionally selected from or comprises any of the nucleotide sequences presented in SEQ ID NO: 155 - SEQ ID NO: 308 in Table 1; wherein the optionally numbered positions 2-19 of the antisense strand in the 5' to 3' direction comprise at least 15 nucleotides from the nucleotides at positions 2-19 of any of the nucleotide sequences presented in SEQ ID NO: 155 - SEQ ID NO: 308 in Table 1, or a nucleotide sequence having less than 3 nucleotide differences from the at least 15 nucleotides; wherein the sense strand optionally comprises at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19 contiguous nucleotides of any of the nucleotide sequences presented in SEQ ID NO: 1 - SEQ ID NO: 154 in Table 1, or a nucleotide sequence having no more than 3 nucleotide differences from the contiguous nucleotides; wherein the sense strand optionally comprises at least 15, at least 16, at least 17, at least 18, at least 19 contiguous nucleotides of any of the nucleotide sequences presented in SEQ ID NO: 1 - SEQ ID NO: 154 in Table 1; wherein the sense strand is optionally selected from or comprises any of the nucleotide sequences presented in SEQ ID NO: 1 - SEQ ID NO: 154 in Table 1.

3. The double-stranded oligonucleotide of claim 2, wherein each nucleotide in the double-stranded oligonucleotide is independently selected from unmodified or modified nucleotides; wherein it is not necessary that essentially all nucleotides of the sense strand or the antisense strand are selected from modified nucleotides; in this case, it is not necessary that all nucleotides of the sense strand and the antisense strand are selected from modified nucleotides; wherein each of the modified nucleotides is optionally independently selected from 2'-halo, 2'-deoxy, 2'-O–(CH2) n -R1modified nucleotides or nucleotide analogs; wherein the nucleotide analogs are selected from one or more of PNA, MNA, BNA, LNA, GNA, TNA and UNA; n is selected from 0, 1 or 2; R1 is selected from optionally substituted C₁₋₆ alkyl, optionally substituted C₁₋₆ alkoxy or –Si(R 1a )3; each R 1a independently selected from optionally substituted C₁₋₆ alkyl or optionally substituted C₁₋₆ alkoxy; in this case, 2'-O–(CH2) is not required n –R1 is selected from 2'-O–CH3, 2'-O–CH2–O–CH3, 2'-O-TBDMS, 2'-O-TIPS, 2'-O-TOM, 2'-O–CH2–O–CH2–CH3, 2’-O–CH2–O–CH2–CF3, or 2’-O–CH2–CH2–O–CH3; wherein the sense strand or the antisense strand optionally comprises a 3'-overhang having at least 1 nucleotide; wherein the antisense strand optionally comprises a 3' overhang having at least 1 nucleotide; wherein the sense strand or the antisense strand optionally comprises a 3'-overhang having at least 2 nucleotides; wherein the antisense strand optionally comprises a 3' overhang having at least 2 nucleotides; wherein the antisense strand optionally contains a 3'-protrusion having 2 nucleotides; wherein the sense strand and / or the antisense strand optionally independently comprise one or more phosphorothioate bonds; in this case, the semantic strand does not necessarily contain two adjacent phosphorothioate bonds between the terminal nucleotides at the 5' end; in this case, the antisense strand optionally contains two adjacent phosphorothioate bonds between the terminal nucleotides at the 3' and 5' ends, respectively.

4. A double-stranded oligonucleotide according to any one of claims 1 to 3, wherein all nucleotides of the sense strand and all nucleotides of the antisense strand are selected from modified nucleotides; wherein the duplex region formed by the sense strand and the antisense strand is represented by the following formula (I): SS: 5'–(N)a'–(X)p'–(N)b'–(X)q'–(N)c'–(X)r'–(N) d'–3' AS: 3'–(N)a–(X)p–(N)b–(X)q–(N)c–5' (I), where SS represents the sense strand and AS represents the antisense strand; each N is independently selected from 2'-fluoro-modified nucleotides, 2'-O-methyl-modified nucleotides, or 2'-deoxy-modified nucleotides; each X is independently selected from 2'-O-TBDMS, 2'-O-TIPS, 2'-O-TOM, 2'-O–CH2–O–CH2–CH3, 2'-O–CH2–O–CH2–CF3, 2'-O–CH2–CH2–O–CH3; each of a, a', p, p', b, b', q, q', c, c', r', d' independently represents a number of nucleotides, wherein: a' is selected from an integer of 3-8; p' is selected from an integer of 0-3; b' is selected from an integer of 4-13; q' is selected from an integer of 0-4; c' is selected from an integer of 3-9; r' is selected from an integer of 0-3; d' is selected from an integer of 0-9; a is selected from an integer of 4-7; p is selected from an integer of 0-1; b is selected from an integer of 4-8; q is selected from an integer of 0-4; c is selected from an integer of 6-10; and p', q', r', p, q are not simultaneously 0, and 0 ≤ q' + r' ≤ 4; wherein optionally a' is selected from an integer of 3-8, p' is selected from 0 or 1, b' is selected from an integer of 4-13, q' is selected from 0 or 1, c' is selected from an integer of 3-9; r' is selected from 0 or 1, d' is selected from an integer of 1-8, a is selected from an integer of 4-7, p is selected from 1, b is selected from an integer of 4-8, q is selected from 0 or 1, c is selected from an integer of 6-10; wherein each N is optionally independently selected from 2'-O-methyl-modified nucleotides, 2'-fluoro-modified nucleotides; wherein each X is optionally independently selected from 2'-O-methyl-modified nucleotides, 2'-O-MOE (methoxyethyl)-modified nucleotides, 2'-O-TBDMS-modified nucleotides, 2'-O-TIPS-modified nucleotides, 2'-O-TOM-modified nucleotides, 2'-O–CH2–O–CH2–CH3-modified nucleotides, 2'-O–CH2–O–CH2–CF3-modified nucleotides; wherein each X is preferably independently selected from 2'-O-methyl-modified nucleotides, 2'-O-MOE-modified nucleotides, 2'-O–CH2–O–CH2–CH3-modified nucleotides, 2'-O–CH2–O–CH2–CF3-modified nucleotides; wherein the duplex region optionally comprises at least one 2'-O-methoxyethyl-modified nucleotide or 2'-O-ethoxymethyl-modified nucleotide.

5. A double-stranded oligonucleotide according to claim 4, wherein in the direction from the 5'-end to the 3'-end at least four nucleotides at positions 2, 6, 9, 12, 14 and 16 of the antisense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides or 2'-O-methoxyethyl-modified nucleotides; wherein, optionally in the direction from the 5'-end to the 3'-end, at least five nucleotides at positions 2, 6, 9, 12, 14 and 16 of the antisense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides or 2'-O-methoxyethyl-modified nucleotides; wherein, optionally in the direction from the 5'-end to the 3'-end, arbitrary five nucleotides at positions 2, 6, 9, 12, 14 and 16 of the antisense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides or 2'-O-methoxyethyl-modified nucleotides; wherein, optionally in the direction from the 5'-end to the 3'-end, the nucleotides at positions 2, 6, 9, 14 and 16 of the antisense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides or 2'-O-methoxyethyl-modified nucleotides; wherein the antisense strand optionally comprises at least one 2'-O-methoxyethyl-modified nucleotide; wherein, optionally in the direction from the 5'-end to the 3'-end, the nucleotide at position 15 of the antisense strand is selected from a 2'-O-methoxyethyl-modified nucleotide; at least four nucleotides at positions 2, 6, 9, 12, 14 and 16 are selected from 2'-fluoro-modified nucleotides; and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; wherein, optionally in the direction from the 5'-end to the 3'-end, the nucleotide at position 15 of the antisense strand is selected from a 2'-O-methoxyethyl-modified nucleotide; at least five nucleotides at positions 2, 6, 9, 12, 14 and 16 are selected from 2'-fluoro-modified nucleotides; and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; wherein, optionally in the direction from the 5'-end to the 3'-end, the nucleotide at position 15 of the antisense strand is selected from a 2'-O-methoxyethyl-modified nucleotide; arbitrary five nucleotides at positions 2, 6, 9, 12, 14 and 16 are selected from 2'-fluoro-modified nucleotides; and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; wherein, optionally in the direction from the 5'-end to the 3'-end, the nucleotide at position 15 of the antisense strand is selected from a 2'-O-methoxyethyl-modified nucleotide; the nucleotides at positions 2, 6, 9, 14 and 16 are selected from 2'-fluoro-modified nucleotides; and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; wherein, optionally in the direction from the 5'-end to the 3'-end, the nucleotide at position 15 of the antisense strand is selected from a 2'-O-methoxyethyl-modified nucleotide; the nucleotides at positions 2, 6, 12, 14 and 16 are selected from 2'-fluoro-modified nucleotides; and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides.

6. A double-stranded oligonucleotide according to claim 4, wherein in the direction from the 5'-end to the 3'-end at least three nucleotides in positions 7-10 of the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides in the remaining positions are selected from 2'-O-methyl-modified nucleotides; wherein, optionally in the direction from the 5'-end to the 3'-end, the nucleotides in positions 7–10 of the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides in the remaining positions are selected from 2'-O-methyl-modified nucleotides.

7. The double-stranded oligonucleotide of claim 4, wherein the antisense strand comprises at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, or at least 21 contiguous nucleotides of any of the modified nucleotide sequences of the antisense strand presented in Table 2, or a nucleotide sequence having no more than 3 nucleotide differences from the contiguous nucleotides; wherein the antisense strand optionally comprises at least 17, at least 18, at least 19, at least 20, or at least 21 contiguous nucleotides of any of the modified nucleotide sequences of the antisense strand presented in Table 2; wherein the antisense strand is optionally selected from or comprises any of the modified antisense strand nucleotide sequences presented in Table 2; wherein the sense strand optionally comprises at least 15, at least 16, at least 17, at least 18, at least 19 contiguous nucleotides of any of the modified nucleotide sequences of the sense strand presented in Table 2, or a nucleotide sequence that differs from the contiguous nucleotides by 1, 2 or 3 nucleotides; wherein the sense strand of the double-stranded oligonucleotide is optionally selected from or comprises any of the modified nucleotide sequences of the sense strand presented in Table 2; wherein, optionally, the sense strand of the double-stranded oligonucleotide is selected from or comprises any of the modified nucleotide sequences of the sense strand presented in Table 2; and the antisense strand is selected from or comprises any of the modified nucleotide sequences of the antisense strand presented in Table 2.

8. A double-stranded oligonucleotide according to any one of claims 1 to 7, selected from siRNA.

9. A double-stranded oligonucleotide conjugate comprising a double-stranded oligonucleotide according to any one of claims 1 to 8 and one or more ligands capable of binding to a cellular receptor; wherein the ligand is optionally conjugated to the sense strand and / or the antisense strand; wherein the ligand is optionally conjugated to the 3'-end and / or 5'-end of the sense strand; in this case, the ligand is optionally conjugated to the 3' end of the sense strand; in this case, the cellular receptor is not necessarily selected from the asialoglycoprotein receptor; wherein the ligand optionally contains galactose or a galactose cluster.

10. The conjugate of claim 9, wherein the ligand is selected from the structure of formula (101), or the corresponding isomer, or the corresponding pharmaceutically acceptable salt: , Where represents the conjugation site between the ligand and the sense strand or antisense strand; m is selected from 1, 2, 3 or 4; each Z is independently selected from hydroxyl or mercapto; each p is independently selected from 1, 2, or 3; each q is independently selected from 1, 2, or 3; each R is independently selected from H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy; each L is independently selected from optionally substituted C2–C20 alkylene or ; R La and R Lb are independently selected from optionally substituted C1-C10 alkylene, and k is selected from 1, 2, 3, 4 or 5; each Y is independently selected from O, S or NH; optionally m is selected from 1, 2 and 3; optionally Z is selected from hydroxyl; optionally p is selected from 1; optionally q is selected from 1; optionally R is selected from H; optionally each L is independently selected from C1–C10 alkylene or ; where R La and R Lb are independently selected from C1-C5 alkylene, and k is 1, 2, or 3; optionally k is selected from 1; it is not necessary that each L is independently selected from , , , , , , , , , , , , , or ; optionally Y is selected from O.

11. The conjugate of claim 9, wherein the ligand is selected from the following structure, or the corresponding isomer, or the corresponding pharmaceutically acceptable salt: .

12. The conjugate of claim 9, wherein the ligand is selected from the structure shown below, or the corresponding isomer, or the corresponding pharmaceutically acceptable salt: , Where represents the conjugation site between the ligand and the sense strand or antisense strand.

13. A composition comprising any of the following: (I) a double-stranded oligonucleotide according to any one of claims 1 to 8; and / or (II) a conjugate according to any one of claims 9 to 12; optionally, the composition is a pharmaceutical composition; optionally, the composition further comprises a pharmaceutically acceptable excipient or auxiliary agent.

14. Use of any of the following: (I) a double-stranded oligonucleotide according to any one of claims 1 to 8; and / or (II) a conjugate according to any one of claims 9 to 12; and / or (III) composition according to claim 13, in obtaining a medicinal product for the prevention and / or treatment of a disease or disorder mediated by the INHBE gene; wherein the optional disease or disorder includes, but is not limited to, the presence of or risk of developing a metabolic disorder, type 2 diabetes mellitus, obesity, elevated triglycerides, lipodystrophy, liver inflammation, fatty liver disease, hypercholesterolemia, elevated liver enzymes, non-alcoholic steatohepatitis (NASH), cardiovascular disease, cardiomyopathy, hypertension, and / or heart failure.

15. A method of reducing the expression or activity of an INHBE gene, comprising contacting a cell with any of the following: (I) a double-stranded oligonucleotide according to any one of claims 1 to 8; and / or (II) a conjugate according to any one of claims 9 to 12; and / or (III) composition according to claim 13.

16. A method for the prevention and / or treatment of a disease or disorder mediated by the INHBE gene, comprising administering to a subject a pharmaceutically acceptable amount of any of the following: (I) a double-stranded oligonucleotide according to any one of claims 1 to 8; and / or (II) a conjugate according to any one of claims 9 to 12; and / or (III) composition according to claim 13.