Inhibitors of expression and / or function

Abstract

The present invention relates to inhibitors, compositions containing inhibitors, and their use in the treatment or prevention of HFrEF-related diseases such as ischemic heart disease.

Claims

1. An inhibitor of NR3C2 expression and / or function, wherein the inhibitor is conjugated to one or more ligand moieties.

2. The inhibitor according to claim 1, which is an siRNA oligomer.

3. An inhibitor of the expression and / or function of NR3C2, which is an siRNA oligomer.

4. The inhibitor according to claim 3, wherein the inhibitor comprises one or more ligand-conjugated siRNA oligomers.

5. Inhibitors of NR3C2 expression and / or function, such as the inhibitors described in claims 1 to 4, for use in the treatment of HFrEF, particularly myocardial infarction and / or symptoms thereof, such as ischemic heart disease.

6. The inhibitor according to claim 1, 2, or 4, wherein the one or more ligand moieties comprise one or more GalNAc ligands, or further comprise one GalNAc ligand derivative.

7. The inhibitor for use according to claim 1, 2, or 4, wherein the one or more ligand moieties comprise one or more GalNAc ligand derivatives.

8. The inhibitor or inhibitor for use according to one or more prior claims, wherein the target of the inhibitor is NR3C2.

9. It is an siRNA oligomer having a first strand and a second strand, i) The first strand of the siRNA has a length of 15 to 30 nucleosides, preferably 19 to 25 nucleosides, more preferably 23 or 25 nucleosides; even more preferably 23 nucleosides; and / or ii) The second strand of siRNA has a length of 15 to 30 nucleosides, preferably 19 to 25 nucleosides, more preferably 21 nucleosides. An inhibitor or inhibitor for use according to one or more prior claims.

10. The inhibitor or inhibitor for use according to claim 9, wherein the second sense strand further comprises one or more debased nucleosides in the terminal region of the second strand, the debased nucleosides being connected to adjacent nucleosides via reverse internucleoside linkages.

11. The second chain is, i. Two or more debase nucleosides in the terminal region of the second chain; and / or ii Two or more debase nucleosides located in either the 5' or 3' terminal region of the second chain; and / or iii Two or more debasic nucleosides located in either the 5' or 3' terminal region of the second chain, which are present as protrusions as described herein; and / or iv Two or more consecutive debasic nucleosides in the terminal region of the second chain, preferably one such debasic nucleoside being a terminal nucleoside, and / or v Two or more consecutive debasic nucleosides located in either the 5' or 3' terminal region of the second chain, preferably one such debasic nucleoside being a terminal nucleoside located in either the 5' or 3' terminal region of the second chain, and / or vi. Reverse internucleoside linkages connecting at least one debasic nucleoside to an adjacent base nucleoside in the terminal region of the second chain, and / or vii Reverse internucleoside linkages, in which at least one debasic nucleoside is connected to an adjacent base nucleoside in either the 5' or 3' terminal region of the second chain, and / or viiii A debased nucleoside as the second-to-last nucleoside connected via reverse linkage to a nucleoside that is not the terminal nucleoside (referred to herein as the third-to-last nucleoside), and / or ix When the chain is read in the direction toward its end, the debasal nucleosides are two terminal nucleosides connected via a 5'-3' linkage. x When the chain is read in the direction toward the terminal containing the aforementioned terminal nucleoside, the debased nucleosides are two terminal nucleosides connected via a 3'-5' linkage. xi A debased nucleoside at the two terminal positions, wherein the second to last nucleoside is connected to the third to last nucleoside via a reverse linkage, and the reverse linkage is a 5-5' reverse linkage or a 3'-3' reverse linkage, xii A debased nucleoside at the two terminal positions, wherein the second to last nucleoside is connected to the third to last nucleoside via the reverse linkage, (1) The reverse linkage is a 5-5' reverse linkage, and the linkage between the terminal debasing nucleoside and the second-to-last debasing nucleoside is 3'5' when read toward the terminal containing the terminal debasing nucleoside and the second-to-last debasing nucleoside, or (2) The reverse linkage is a 3-3' reverse linkage, and the linkage between the terminal debasing nucleoside and the second-to-last debasing nucleoside is either 5'3' when read toward the terminal containing the terminal debasing nucleoside and the second-to-last debasing nucleoside, debasing nucleoside The inhibitor or inhibitor for use according to claim 10, comprising:

12. The inhibitor according to claim 10 or 11, or an inhibitor for use, wherein the reverse nucleoside linkage is located in the distal terminal region of the second chain from the 5' terminal region, or in the distal terminal region of the second chain from the 3' terminal region.

13. The inhibitor or inhibitor for use according to any one of claims 10 to 12, wherein the reverse nucleoside linkage is a 3'3 reverse linkage.

14. The inhibitor or inhibitor for use according to any one of claims 10 to 12, wherein the reverse nucleoside linkage is a 5'5 reverse linkage.

15. The inhibitor or inhibitor for use according to any one of claims 9 to 14, wherein one or more nucleosides in the first chain and / or the second chain are modified to form a modified nucleoside.

16. The inhibitor or inhibitor for use according to claim 15, wherein the modification is a modification of the 2'-OH group of ribose sugar, and is optionally selected from 2'-Me or 2'-F modifications.

17. The inhibitor or inhibitor for use according to claim 15 or 16, wherein the first chain contains 2'-F at any of the 14th, 2nd, or 6th positions, or any combination thereof, counting from the 1st position of the first chain.

18. The inhibitor or inhibitor for use according to any one of claims 15 to 17, wherein the second chain includes a 2'-F modification at the 7th and / or 9th and / or 11th and / or 13th positions, counting from the 1st position of the second chain.

19. The inhibitor or inhibitor for use according to any one of claims 15 to 18, wherein the first and second chains each comprise 2'-Me and 2'-F modifications.

20. An inhibitor or inhibitor for use according to any one of claims 15 to 19, wherein the siRNA preferably comprises at least one thermal destabilization modification at one or more positions 1 to 9 of the first strand counting from position 1 of the first strand, and / or at one or more positions of the second strand aligned with positions 1 to 9 of the first strand, the destabilization modification being selected from modified unlocked nucleic acids (IMUNAs) and glycol nucleic acids (GNAs), preferably glycol nucleic acids.

21. The inhibitor or inhibitor for use according to claim 20, wherein the siRNA comprises at least one thermal destabilization modification at position 7 of the first strand, counting from position 1 of the first strand.

22. An inhibitor or inhibitor for use according to any one of claims 15 to 21, wherein the siRNA comprises three or more 2'-F modifications at positions 7 to 13 of the second strand, counting from position 1 of the second strand, for example, four, five, six, or seven 2'-F modifications at positions 7 to 13 of the second strand.

23. An inhibitor or inhibitor for use according to any one of claims 15 to 22, wherein the siRNA, the second strand comprises at least three, for example, four, five, or six 2'-Me modifications at positions 1 to 6 of the second strand, counting from position 1 of the second strand.

24. The inhibitor or inhibitor for use according to any one of claims 15 to 23, wherein the first strand is an siRNA, and preferably includes at least five 2'-Me serial modifications in the 3'-terminal region, including the terminal nucleoside of the 3'-terminal region, or within at least one or two nucleosides from the terminal nucleoside of the 3'-terminal region.

25. An inhibitor or inhibitor for use according to any one of claims 15 to 24, wherein the first strand is a siRNA, and preferably the 3' terminal region includes seven 2'-Me serial modifications, including a terminal nucleoside in the 3' terminal region.

26. The inhibitor or inhibitor for use according to any one of claims 15 to 25, wherein the siRNA oligomer further comprises one or more phosphorothioate nucleoside linkages.

27. The inhibitor or inhibitor for use according to claim 26, wherein each of the one or more phosphorothioate nucleoside links is located between at least three consecutive positions in the 5' or 3' near-terminal region of the second chain, and the near-terminal region is preferably adjacent to the terminal region in which the one or more debased nucleosides of the second chain are located as described in claim 10.

28. The inhibitor or inhibitor for use according to claim 26 or 27, wherein each of the one or more phosphorothioate nucleoside linkages is located between at least three consecutive positions in the 5' and / or 3' terminal region of the first chain, and preferably the terminal position of the 5' and / or 3' terminal region of the first chain is attached to its adjacent position by the phosphorothioate nucleoside linkage.

29. The inhibitor or inhibitor for use according to any one of claims 9 to 28, wherein the oligomer is siRNA, and the second strand of the siRNA is directly or indirectly conjugated to one or more ligand moieties, the ligand moieties typically located in the terminal region of the second strand, preferably in its 3' terminal region.

30. The ligand portion is i) One or more GalNAc ligands, and / or ii) One or more GalNAc ligand derivatives, and / or iii) One or more GalNAc ligands and / or GalNAc ligand derivatives conjugated to the siRNA via a linker. The inhibitor or inhibitor for use according to claim 29, comprising:

31. The inhibitor or inhibitor for use according to claim 30, wherein the one or more GalNAc ligands and / or GalNAc ligand derivatives are conjugated, preferably directly or indirectly, to the 5' or 3' terminal region of the second chain of the siRNA oligomer.

32. The ligand portion is 【Chemistry 1】 The inhibitor or inhibitor for use according to claim 30 or 31, comprising:

33. structure: 【Chemistry 2】 It has, During the ceremony, R １ Each instance is independently selected from the group consisting of hydrogen, methyl, and ethyl; R ２ is hydrogen, hydroxyl, -OC １～３ Alkyl, -C(=O)OC １～３ Selected from the group consisting of alkyl, halo, and nitro, X １ and X ２ In each occurrence, it is independently selected from the group consisting of methylene, oxygen, and sulfur. m is an integer from 1 to 6. n is an integer between 1 and 10. q, r, s, t, and v are independent integers between 0 and 4, however, (i) q and r cannot both be zero at the same time, (ii) s, t, and v must all be zero at the same time. Z is an oligomer. The inhibitor or inhibitor for use according to claim 30 or 31.

34. structure: 【Transformation 3】 It has, in the formula, r and s are integers independently selected from 1 to 16; and Z is an oligomer. The inhibitor or inhibitor for use according to claim 30 or 31.

35. One or more inhibitors or inhibitors for use according to a preceding claim, formulated as a pharmaceutical composition having an excipient and / or a carrier.

36. A pharmaceutical composition comprising one or more inhibitors according to a prior claim, in combination with a pharmaceutically acceptable excipient or carrier.

37. A pharmaceutical composition comprising one or more inhibitors according to a preceding claim, in combination with a pharmaceutically acceptable excipient or carrier, for use in the treatment of HFrEF such as ischemic heart disease, particularly myocardial infarction and / or symptoms thereof.

38. The use of NR3C2 as a target for identifying one or more therapeutic agents for the treatment of HFrEF, particularly myocardial infarction and / or its symptoms, such as ischemic heart disease.

39. A method for treating HFrEF, particularly myocardial infarction and / or symptoms thereof, such as ischemic heart disease, comprising administering an NR3C2 inhibitor, such as one or more inhibitors described in the preceding claims, to a patient.

40. NR3C2 for use as a biomarker for HFrEF and related symptoms of ischemic heart disease, particularly myocardial infarction and / or its symptoms.

41. Typically, NR3C2 is used in in vivo methods to predict susceptibility to HFrEF, particularly myocardial infarction and / or its symptoms, such as ischemic heart disease, by monitoring the sequence and / or expression and / or function levels of NR3C2 in patient samples.

42. A method for predicting susceptibility to HFrEF, such as ischemic heart disease, particularly myocardial infarction and / or its symptoms, and a method for selectively treating HFrEF-related diseases, such as ischemic heart disease, particularly myocardial infarction and / or its symptoms, in a patient. (a) Obtain a sample from the patient, (b) To detect the sequence and / or expression and / or function of NR3C2 in the sample obtained from the patient, (c) Predict susceptibility to HFrEF-related diseases such as ischemic heart disease based on the sequence and / or expression and / or function of NR3C2 in the sample obtained from the patient. (d) Preferably, administer an effective dose of an NR3C2 inhibitor to the diagnosed patient. A method that includes this.

43. An inhibitor or composition according to any of the preceding claims in the preparation of a pharmaceutical for use in the treatment of HFrEF, ischemic heart disease, particularly myocardial infarction and / or symptoms thereof.

44. The inhibitor or composition for use according to any of the preceding claims, wherein the inhibitor or composition is administered after myocardial infarction.

45. A nucleic acid for inhibiting the expression of NR3C2, comprising a double-stranded region including a first strand and a second strand at least partially complementary to the first strand, wherein the first strand is (i) at least partially complementary to the portion of RNA transcribed from the NR3C2 gene, and (ii) A nucleic acid comprising at least 17 consecutive nucleosides that differ by 0 or 1 nucleoside from any one of the first strand sequences listed in Table 2.

46. A nucleic acid for inhibiting the expression of NR3C2, comprising a double-stranded region including a first strand and a second strand at least partially complementary to the first strand, wherein the first strand is (i) at least partially complementary to the portion of RNA transcribed from the NR3C2 gene, and (ii) A nucleic acid comprising at least 17 consecutive nucleosides that differ by 0 or 1 nucleosides from any one of the first chain modification sequences listed in Table 3.

47. The nucleic acid according to claim 45 or 46, wherein the first strand comprises nucleosides 2 to 18 of any one sequence described in claim 45 or 46, and in particular the first strand comprises nucleosides 2 to 18 of any one sequence defined in Table 2 or 3.

48. The nucleic acid according to claim 45, wherein the second strand comprises a nucleoside sequence of at least 17 consecutive nucleosides that differ by 0 or 1 nucleosides from any one of the second strand sequences listed in Table 2, and the second strand has a region that is at least 85% complementary to the first strand across the 17 consecutive nucleosides.

49. The nucleic acid according to claim 46, wherein the second chain comprises a nucleoside sequence of at least 17 consecutive nucleosides that differ by 0 or 1 nucleosides from any one of the second chain modification sequences listed in Table 4, and the second chain has a region that is at least 85% complementary to the first chain across the 17 consecutive nucleosides.

50. The nucleic acid according to claim 45, wherein the first chain comprises any one of the sequences of the first chain listed in Table 2.

51. The nucleic acid according to claim 46, wherein the first chain comprises any one of the modification sequences of the first chain listed in Table 3.

52. The nucleic acid according to claim 48, wherein the second chain comprises any one of the second chain sequences listed in Table 2.

53. The nucleic acid according to claim 49, wherein the second chain comprises any one of the second chain modification sequences listed in Table 4.

54. The nucleic acid according to any one of claims 45 to 53, wherein the first chain has a length of 17 to 30 nucleosides, preferably 19 to 25 nucleosides, more preferably 19 or 23 nucleosides.

55. The nucleic acid according to any one of claims 45 to 54, wherein the second chain has a length of 17 to 30 nucleosides, preferably 19 to 25 nucleosides, and more preferably 19, 21, or 23 nucleosides.

56. The nucleic acid according to any one of claims 45 to 55, wherein the double-stranded region of the nucleic acid has a nucleoside length between 17 and 30, more preferably 19, 21, or 23.

57. The nucleic acid according to any one of claims 45 to 56, wherein the complementary region between the first strand and the portion of RNA transcribed from the NR3C2 gene is between 17 and 30 nucleoside lengths.

58. The nucleic acid according to any one of claims 45 to 57, further comprising one or more single-stranded nucleoside protrusions, optionally the protrusions being located on the first or second strand, preferably at the 3' end of the first or second strand, and / or the protrusions comprising 1 to 4 nucleosides, more preferably 2 nucleosides.

59. The nucleic acid according to any one of claims 45 to 58, which is an siRNA oligonucleoside.

60. The second chain contains two consecutive debased nucleosides in the 5' terminal region of the second chain, one of which is the terminal nucleoside of the 5' terminal region of the second chain, and the other debased nucleoside is the second to last nucleoside of the 5' terminal region of the second chain. (a) The second to last debasic nucleoside is connected to the adjacent first base nucleoside in the adjacent 5' near-terminal region via an inverse nucleoside linkage, (b) The reverse connection is a 5-5' reverse connection, and (c) The linkage between the terminal debasing nucleoside and the second-to-last debasing nucleoside is 3'5' when read toward the terminal including the terminal debasing nucleoside and the second-to-last debasing nucleoside. The nucleic acid according to claim 49 or 53.

61. (i) The first chain and the second chain each have a length of 23 nucleosides, (ii) The two phosphorothioate nucleoside links are each located between three consecutive positions in the 5' near-terminal region of the second chain, the first phosphorothioate nucleoside link is located between the adjacent first base nucleoside of (a) and the adjacent second base nucleoside in the 5' near-terminal region of the second chain, the second phosphorothioate nucleoside link is located between the adjacent second base nucleoside and the adjacent third base nucleoside in the 5' near-terminal region of the second chain, (iii) The two phosphorothioate nucleoside links are each located between three consecutive positions in both the 5' and 3' terminal regions of the first chain, and each terminal nucleoside in each of the 5' and 3' terminal regions of the first chain is attached to the second-to-last adjacent nucleoside of 5' and 3', respectively, by phosphorothioate nucleoside links, and each second-to-last nucleoside of the first 5' and 3' is attached to the third-to-last adjacent nucleoside of 5' and 3', respectively, by phosphorothioate nucleoside links, and (iv) The second strand of the nucleic acid is directly or indirectly conjugated to one or more ligand moieties in the 3' terminal region of the second strand. The nucleic acid according to claim 50.

62. The two consecutive reverse debase nucleosides in the 5' terminal region of the second chain form the following 5' terminal motif 【Chemistry 4】 It exists as, During the ceremony, T represents the 2'Me ribose modification, B represents the nucleoside base of the two first base nucleosides in the 5' terminal region of the second chain, and Z represents the remaining 19 consecutive base nucleosides of the second chain. The nucleic acid according to claim 50 or 51.

63. The nucleic acid according to any one of claims 45 to 62, wherein the nucleic acid is directly or indirectly conjugated to one or more ligand moieties, and optionally the ligand moieties are located in the terminal region of the second chain, preferably in its 3' terminal region.

64. The ligand portion is (i) one or more N-acetylgalactosamine (GalNAc) ligands, and / or (ii) One or more N-acetylgalactosamine (GalNAc) ligand derivatives The nucleic acid according to claim 63, comprising:

65. The nucleic acid according to claim 64, wherein the one or more GalNAc ligands and / or GalNAc ligand derivatives are directly or indirectly conjugated to the 5' or 3' terminal region, preferably the 3' terminal region, of the second chain of the nucleic acid.

66. structure: 【Transformation 5】 Includes During the ceremony, R1 is independently selected in each occurrence from the group consisting of hydrogen, methyl, and ethyl; R2 is selected from the group consisting of hydrogen, hydroxyl, -OC1-3 alkyl, -C(=O)OC1-3 alkyl, halo, and nitro. X1 and X2 are independently selected in each occurrence from the group consisting of methylene, oxygen, and sulfur. m is an integer from 1 to 6. n is an integer between 1 and 10. q, r, s, t, and v are independent integers between 0 and 4, however, (i) q and r cannot both be zero at the same time, (ii) s, t, and v must all be zero at the same time. Z is the oligonucleoside portion. The nucleic acid according to any one of claims 63 to 65.

67. structure: 【Transformation 6】 The oligonucleotide comprises the continuous nucleoside of the second chain. The nucleic acid according to claim 66.

68. structure: 【Transformation 7】 Includes, During the ceremony, r and s are integers independently selected from 1 to 16, and Z is the oligonucleoside portion. The nucleic acid according to any one of claims 63 to 65.

69. structure 【Transformation 8】 The nucleic acid according to claim 68, comprising, wherein the oligonucleotide represents the continuous nucleoside of the second chain.

70. The nucleic acid according to claim 67 or 69, wherein the structure is conjugated to the 3' terminal region of the second chain.

71. The nucleic acid according to claims 62, 67, and 70.

72. The nucleic acid according to claims 62, 69, and 70.

73. A pharmaceutical composition comprising the nucleic acid according to any one of claims 45 to 72, in combination with a pharmaceutically acceptable excipient or carrier.

74. A nucleic acid or pharmaceutical composition according to any one of claims 45 to 73, for use in therapeutic purposes.

75. A nucleic acid or pharmaceutical composition according to any one of claims 45 to 74, for use in the treatment of HFrEF such as ischemic heart disease, particularly myocardial infarction and / or symptoms thereof.

76. A nucleic acid or pharmaceutical composition according to claim 75, administered after a myocardial infarction.

77. The nucleic acid according to claim 50, wherein the first strand comprises the following sequences: SEQ ID NO: 228, SEQ ID NO: 238, SEQ ID NO: 243, SEQ ID NO: 235, SEQ ID NO: 245, and SEQ ID NO: 250, preferably one of SEQ ID NO: 238 or SEQ ID NO:

245.

78. The nucleic acid according to claim 51, wherein the first strand comprises the following sequences: SEQ ID NO: 674, SEQ ID NO: 684, SEQ ID NO: 689, SEQ ID NO: 681, SEQ ID NO: 691, and SEQ ID NO: 696, preferably one of SEQ ID NO: 684 or SEQ ID NO:

691.

79. The nucleic acid according to claim 52, wherein the second strand comprises the following sequences: SEQ ID NO: 451, SEQ ID NO: 461, SEQ ID NO: 466, SEQ ID NO: 458, SEQ ID NO: 468, and SEQ ID NO: 473, preferably one of SEQ ID NO: 461 or SEQ ID NO:

468.

80. The nucleic acid according to claim 53, wherein the second strand comprises the following sequences: SEQ ID NO: 897, SEQ ID NO: 907, SEQ ID NO: 912, SEQ ID NO: 904, SEQ ID NO: 914, and SEQ ID NO: 919, preferably one of SEQ ID NO: 907 or SEQ ID NO:

914.

81. The nucleic acid according to claim 45 or 48, comprising a first strand and a second strand, each containing, consisting of, or essentially consisting of, any one of the following combinations of the first and second sequences and a nucleoside sequence that differs by 0 or 1 nucleoside. Table 1 Preferably Table 2 or Table 3

82. The nucleic acid according to claim 46 or 49, comprising a first strand and a second strand, each containing, consisting of, or essentially consisting of, any one of the following combinations of the first and second sequences and a nucleoside sequence that differs by 0 or 1 nucleoside. Table 4 Preferably Table 5 or Table 6

Images