siRNA, siRNA-based conjugate, pharmaceutical composition based on them and their use

Claims

1. An siRNA for suppressing the expression of angiotensinogen, comprising a sense strand and an antisense strand, wherein the antisense strand comprises at least 19 contiguous nucleotides that differ by no more than 3 nucleotides from any of the antisense strand sequences selected from SEQ ID NO: 113, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 109, 111, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, and 143; the antisense strand contains no more than 27 contiguous nucleotides, and the number of nucleotides in the sense strand is 16-25; and where the region where the sense strand and antisense strand are complementarily linked contains at least 15 contiguous nucleotides.

2. The siRNA of claim 1, wherein the antisense strand comprises a terminal overhang of 1-5 nucleotides in length at the 3' end.

3. The siRNA of claim 1 or 2, wherein the sense strand contains at most one mismatched base with the antisense strand, or the sense strand does not contain any mismatched bases with the antisense strand.

4. The siRNA of any one of claims 1 to 3, wherein the antisense strand comprises a sequence as represented by any SEQ ID NO selected from: 113, 92, 94, 104, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 60, 66, 68, 96, 98, 100, and 102.

5. The siRNA of any one of claims 1 to 4, wherein the siRNA is an unmodified duplex selected from any of duplex 26, duplex 24, duplex 25, duplex 27, and duplexes 1 to 23.

6. siRNA according to any one of claims 1-5, wherein all nucleotides in the sense strand and / or antisense strand are modified nucleotides or nucleotide analogs, wherein the modified nucleotides are at least one nucleotide selected from 2'-methoxynucleotides, 2'-fluoronucleotides, 2'-deoxynucleotides, 2'-methoxyethylnucleotides, 2'-aminonucleotides, 2'-alkylnucleotides, 3'-methoxynucleotides, (E)-vinylphosphonate-modified nucleotides, (Z)-vinylphosphonate-modified nucleotides, phosphorothioate-modified nucleotides, phosphorothioate-modified nucleotides in the Sp configuration, phosphorothioate-modified nucleotides in the Rp configuration, or mesylphosphoramidate-modified nucleotides, and the nucleotide analogs include closed nucleic acids, open nucleic acids, or glycerol nucleic acids; and / or at least one of the bonds between the nucleotide at position 1 and the nucleotide at position 2 at the 5' end of the sense strand, between the nucleotide at position 2 and the nucleotide at position 3 at the 5' end of the sense strand, between the nucleotide at position 1 and the nucleotide at position 2 at the 3' end of the antisense strand, between the nucleotide at position 2 and the nucleotide at position 3 at the 3' end of the antisense strand, between the nucleotide at position 1 and the nucleotide at position 2 at the 5' end of the antisense strand, and between the nucleotide at position 2 and the nucleotide at position 3 at the 5' end of the antisense strand is a phosphorothioate bond, a phosphorothioate bond in the Rp configuration, or a phosphorothioate bond in the Sp configuration.

7. The siRNA of any one of claims 1 to 6, wherein four, five, six or seven nucleotides in the antisense strand selected from nucleotides at positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 14, 16, 18, 20, 21 and optionally 22 in the direction from the 5' end to the 3' end of the antisense strand are 2'-fluoronucleotides, and all of the nucleotides at positions 2, 6, 14 and 16 are 2'-fluoronucleotides.

8. The siRNA of any one of claims 1 to 7, wherein the antisense strand comprises the following modified nucleotides: (1) the nucleotides at positions 2, 6, 7, 14, 16, and 22 in the 5' to 3' direction of the antisense strand are 2'-fluoronucleotides; the nucleotides at other positions of the antisense strand may be 2'-methoxynucleotides; the nucleotide at position 1 in the 5' to 3' direction of the antisense strand further comprises an (E)-VP modification; (2) the nucleotides at positions 2, 6, 14, and 16 in the 5' to 3' direction of the antisense strand are 2'-fluoronucleotides; one or two of the nucleotides at positions 4, 8, 9, 10, 18, and 20 in the 5' to 3' direction of the antisense strand may additionally be 2'-fluoronucleotides; the nucleotides at other positions of the antisense strand are 2'-methoxynucleotides; (3) the nucleotides at positions 2, 6, 14, and 16 in the 5' to 3' direction of the antisense strand and the nucleotide at position 2 in the 3' to 5' direction of the antisense strand are 2'-fluoronucleotides; one or two of the nucleotides at positions 3, 4, 5, 7, 8, 9, 10, 12, and 20 in the 5' to 3' direction of the antisense strand may further be 2'-fluoronucleotides; the nucleotides at other positions of the antisense strand are 2'-methoxynucleotides; the nucleotide at position 1 in the 5' to 3' direction of the antisense strand may further comprise an (E)-VP modification; (4) the nucleotides at positions 2, 6, 7, 14, and 16 in the 5' to 3' direction of the antisense strand and the nucleotide at position 2 in the 3' to 5' direction of the antisense strand are 2'-fluoronucleotides; the nucleotide at position 10 in the 5' to 3' direction of the antisense strand may further be a 2'-fluoronucleotide; the nucleotides at other positions of the antisense strand are 2'-methoxynucleotides; the nucleotide at position 1 in the 5' to 3' direction of the antisense strand further comprises an (E)-VP modification; (5) the nucleotides at positions 2, 14, and 16 in the 5' to 3' direction of the antisense strand and the nucleotide at position 2 in the 3' to 5' direction of the antisense strand are 2'-fluoronucleotides; one of the nucleotides at positions 4, 8, and 9 in the 5' to 3' direction of the antisense strand may further be a 2'-fluoronucleotide; the nucleotides at other positions of the antisense strand are 2'-methoxynucleotides; (6) the nucleotides at positions 2, 6, and 14 in the 5' to 3' direction of the antisense strand and the nucleotide at position 2 in the 3' to 5' direction of the antisense strand are 2'-fluoronucleotides; one of the nucleotides at positions 4, 8, 9, and 18 in the 5' to 3' direction of the antisense strand may further be a 2'-fluoronucleotide; the nucleotides at other positions of the antisense strand are 2'-methoxynucleotides; or (7) the nucleotides at positions 6, 14, and 16 in the 5' to 3' direction of the antisense strand and the nucleotide at position 2 in the 3' to 5' direction of the antisense strand are 2'-fluoronucleotides; one of the nucleotides at positions 4, 8, 9, and 18 in the 5' to 3' direction of the antisense strand may further be a 2'-fluoronucleotide; the nucleotides at other positions of the antisense strand are 2'-methoxynucleotides.

9. The siRNA of any one of claims 1 to 8, wherein the nucleotides at positions 9, 11, 12, 13, and 15 in the 3' to 5' direction of the sense strand are 2'-fluoronucleotides, the nucleotides at other positions of the sense strand are 2'-methoxynucleotides, and the nucleotides at positions 19 and 20, as well as the nucleotides at positions 20 and 21 in the 3' to 5' direction of the sense strand are linked via phosphorothioate groups; and the nucleotides at positions 2, 6, 7, 14, 16, and 22 in the 5' to 3' direction of the antisense strand are 2'-fluoronucleotides, the nucleotides at other positions of the antisense strand are 2'-methoxynucleotides, the nucleotide at position 1 in the 5' to 3' direction of the antisense strand further comprises an (E)-VP modification, and the nucleotides at positions 1 and 2, the nucleotides at positions 2 and 3, the nucleotides at positions 21 and 22, and the nucleotides at positions 22 and 23 in the 5' to 3' direction of the antisense strand are linked via phosphorothioate groups.

10. The siRNA of any one of claims 1 to 9, wherein the siRNA is at least one of the duplexes selected from modified duplex 46 to modified duplex 50, modified duplex 60 to modified duplex 125, modified duplex 129 to modified duplex 130, modified duplex 133 to modified duplex 149, and modified duplex 152 to modified duplex 201.

11. The siRNA of claim 10, wherein the siRNA is at least one of the duplexes selected from modified duplex 46, modified duplex 61 to modified duplex 64, modified duplex 66 to modified duplex 116, modified duplex 117 to modified duplex 125, modified duplex 129 to modified duplex 130, modified duplex 133 to modified duplex 136, modified duplex 155 to modified duplex 157, modified duplex 162, modified duplex 166, modified duplex 171 to modified duplex 172, and modified duplex 181 to modified duplex 198.

12. An siRNA-based conjugate, wherein the siRNA-based conjugate is obtained by conjugating the siRNA of any one of claims 1-11 to a conjugation molecule.

13. The siRNA-based conjugate of claim 12, wherein the conjugated molecule comprises ; or ; or ; or ; or the structure of the conjugate is as follows: , where the conjugation molecule is covalently conjugated to the 3' end of the sense strand of the siRNA in a manner as shown in the formula above.

14. The siRNA conjugate of claim 12 or 13, wherein the siRNA conjugate is selected from at least one of conjugate 46 to conjugate 50, conjugate 60 to conjugate 125, conjugate 129 to conjugate 130, conjugate 133 to conjugate 136, conjugate 150 to conjugate 151, conjugate 155 to conjugate 204; or the siRNA-based conjugate is selected from at least one of conjugate 150, conjugate 151, conjugate 155, conjugate 156, conjugate 157, conjugate 162, conjugate 166, conjugate 171, conjugate 172, conjugate 175, conjugate 181, conjugate 182, conjugate 183, conjugate 184, conjugate 185; or the siRNA-based conjugate is selected from at least one of conjugate 155, conjugate 156, conjugate 157, conjugate 162, conjugate 171, conjugate 172, conjugate 175, conjugate 182, conjugate 183 and conjugate 184.

15. The siRNA-based conjugate of claim 12 or 13, wherein the antisense strand comprises the following sequence in the 5'-3' direction: (E)-VPmU*i2FA*mCmUmCi2FAX2mUmAX3mAmAmGi2FAmAi2FAmAmGmGmX4mG*i2FG*mG, where X2 is mU or i2FU, X3 is mG or i2FG, and X4 is A or U; and / or the semantic thread contains the following sequence in the 5'-3' direction: mC*mX5*mCmCmUmUi2FUmUi2FCi2FUi2FUmCi2FUmAmAmUmGmAmGmUmAGalNAc(L96), where X5 represents A or U.

16. A pharmaceutical composition for the prevention and / or treatment of diseases associated with angiotensinogen dysregulation, comprising an siRNA-based conjugate according to any one of claims 12-15 as an active ingredient and a pharmaceutically acceptable carrier.

17. A method for preventing and / or treating diseases associated with angiotensinogen dysregulation, comprising administering to a subject in need thereof an siRNA-based conjugate according to any one of claims 12-15, wherein the diseases associated with angiotensinogen dysregulation are selected from hypertension or related conditions.

18. The method of claim 17, wherein the hypertension or related conditions are selected from at least one of transient hypertension, essential hypertension, symptomatic hypertension, hypertensive crisis, hypertensive emergency, isolated systolic and diastolic hypertension, pregnancy-related hypertension, diabetic hypertension, resistant hypertension, refractory hypertension, paroxysmal hypertension, renovascular hypertension, Goldblatt's hypertension, ocular hypertension, pulmonary hypertension, portal hypertension, systemic venous hypertension, systolic hypertension and labile hypertension, cardiac disease accompanied by hypertension, and hypertensive nephropathy.

19. Use of an siRNA-based conjugate according to any one of claims 12-15 in the preparation of a medicinal product for the prevention and / or treatment of diseases associated with angiotensinogen dysregulation, wherein the diseases associated with angiotensinogen dysregulation are selected from hypertension or related conditions.

20. The use according to claim 19, wherein the hypertension or related conditions are selected from at least one of transient hypertension, essential hypertension, symptomatic hypertension, hypertensive crisis, emergency state associated with hypertension, isolated systolic and diastolic hypertension, pregnancy-associated hypertension, diabetic hypertension, resistant hypertension, refractory hypertension, paroxysmal hypertension, renovascular hypertension, Goldblatt's hypertension, ocular hypertension, pulmonary hypertension, portal hypertension, systemic venous hypertension, systolic hypertension and labile hypertension, cardiac disease accompanied by hypertension, and hypertensive nephropathy.