US 12,590,304 B2 · Nucleic acid, pharmaceutical composition, conjugate, preparation method, and use

Abstract

An siRNA which inhibits kininogen (KNG) gene expression, a pharmaceutical composition containing the siRNA, and an siRNA conjugate. Each nucleotide in the siRNA is independently a modified or unmodified nucleotide. The siRNA contains a sense strand and an antisense strand. The sense strand contains nucleotide sequence I, nucleotide sequence I having the same length as the nucleotide sequence shown in SEQ ID NO: 1, with no more than three nucleotide differences. The antisense strand contains nucleotide sequence II, nucleotide sequence II having the same length as the nucleotide sequence shown in SEQ ID NO: 2, with no more than three nucleotide differences. The siRNA, the pharmaceutical composition thereof and the siRNA conjugate can effectively treat and/or prevent septicemia.

An siRNA conjugate having a structure as shown by Formula (308):

wherein:
n1 is an integer selected from 1-3, and n3 is an integer selected from 0-4;
each of m1, m2, or m3 is independently an integer selected from 2-10;
each of R10, R11, R12, R13, R14 or R15 is independently H or selected from the group consisting of C1-C10 alkyl, C1-C10 haloalkyl and C1-C10 alkoxy;
and
R3 is a group having a structure as shown by Formula A59:
wherein, E1 is OH, SH or BH2, and Nu is the siRNA;
wherein the siRNA comprises a sense strand and an antisense strand, and each nucleotide in the siRNA is independently a modified or unmodified nucleotide, wherein the sense strand comprises a nucleotide sequence I, and the antisense strand comprises a nucleotide sequence II;
the nucleotide sequence I and the nucleotide sequence II are at least partly reverse complementary to form a double-stranded region;
and the nucleotide sequence I and the nucleotide sequence II are selected from a group of sequences shown in the following i)-vi):
i) the nucleotide sequence I has the same length as and no more than three nucleotide differences from the nucleotide sequence shown in SEQ ID NO:1;
and the nucleotide sequence II has the same length as and no more than three nucleotide differences from the nucleotide sequence shown in SEQ ID NO:2:
(SEQ ID NO:
1)
5′-AAAGUAACAACCAGUUUGZ1-3′;
(SEQ ID NO:
2)
5′-Z2CAAACUGGUUGUUACUUU-3′,
wherein, Z1 is U, and Z2 is A;
and the nucleotide sequence I comprises a nucleotide Z3 at a corresponding site to Z1, the nucleotide sequence II comprises a nucleotide Z4 at a corresponding site to Z2, and Z4 is the first nucleotide from the 5′ terminal of the antisense strand, Z3 is the first nucleotide from the 3′ terminal of the sense strand;
the nucleotide difference between the nucleotide sequence II and the nucleotide sequence shown in SEQ ID NO:
2 comprises a difference at the site of Z4, and Z4 is selected from U, C or G;
ii) the nucleotide sequence I has the same length as and no more than three nucleotide differences from the nucleotide sequence shown in SEQ ID NO:
61;
and the nucleotide sequence II has the same length as and no more than three nucleotide differences from the nucleotide sequence shown in SEQ ID NO:
62:
(SEQ ID NO:
61)
5′-AUUGAACUUUCGAAUUACZ5-3′;
(SEQ ID NO:
62)
5′-Z6GUAAUUCGAAAGUUCAAU-3′,
wherein, Z5 is C, and Z6 is G;
and the nucleotide sequence I comprises a nucleotide Z7 at a corresponding site to Z5, the nucleotide sequence II comprises a nucleotide Z8 at a corresponding site to Z6, and Z8 is the first nucleotide from the 5′ terminal of the antisense strand, Z7 is the first nucleotide from the 3′ terminal of the sense strand;
the nucleotide difference between the nucleotide sequence II and the nucleotide sequence shown in SEQ ID NO:
62 comprises a difference at the site of Z8, and Z8 is selected from A, U or C;
iii) the nucleotide sequence I has the same length as and no more than three nucleotide differences from the nucleotide sequence shown in SEQ ID NO:
121;
and the nucleotide sequence II has the same length as and no more than three nucleotide differences from the nucleotide sequence shown in SEQ ID NO:
122:
(SEQ ID NO:
121)
5′-UCGAAUUACCUACUCAAUZ9-3′;
(SEQ ID NO:
122)
5′-Z10AUUGAGUAGGUAAUUCGA-3′,
wherein, Z9 is U, and Z10 is A;
and the nucleotide sequence I comprises a nucleotide Zu at a corresponding site to Z9, the nucleotide sequence II comprises a nucleotide Z12 at a corresponding site to Z10, and Z12 is the first nucleotide from the 5′ terminal of the antisense strand, Z11 is the first nucleotide from the 3′ terminal of the sense strand:
the nucleotide difference between the nucleotide sequence II and the nucleotide sequence shown in SEQ ID NO:
122 comprises a difference at the site of Z12, and Z12 is selected from U, C or G;
iv) the nucleotide sequence I has the same length as and no more than three nucleotide differences from the nucleotide sequence shown in SEQ ID NO:
181;
and the nucleotide sequence II has the same length as and no more than three nucleotide differences from the nucleotide sequence shown in SEQ ID NO:
182:
(SEQ ID NO:
181)
5′-GAUAAUGCAUACAUCGAUZ13-3′;
(SEQ ID NO:
182)
5′-Z14AUCGAUGUAUGCAUUAUC-3′,
wherein, Z13 is A, and Z14 is U;
and the nucleotide sequence I comprises a nucleotide Z15 at a corresponding site to Z13, the nucleotide sequence II comprises a nucleotide Z16 at a corresponding site to Z14, and Z16 is the first nucleotide from the 5′ terminal of the antisense strand, Z15 is the first nucleotide from the 3′ terminal of the sense strand:
the nucleotide difference between the nucleotide sequence II and the nucleotide sequence shown in SEQ ID NO:
182 comprises a difference at the site of Z16, and Z16 is selected from A, C or G;
v) the nucleotide sequence I has the same length as and no more than three nucleotide differences from the nucleotide sequence shown in SEQ ID NO:
241;
and the nucleotide sequence II has the same length as and no more than three nucleotide differences from the nucleotide sequence shown in SEQ ID NO:
242:
(SEQ ID NO:
241)
5′-GAAUAACGCAACUUUCUAZ17-3′;
(SEQ ID NO:
242)
5′-Z18UAGAAAGUUGCGUUAUUC-3′,
wherein, Z17 is U, and Z18 is A;
and the nucleotide sequence I comprises a nucleotide Z19 at a corresponding site to Z17, the nucleotide sequence II comprises a nucleotide Z20 at a corresponding site to Z18, and Z20 is the first nucleotide from the 5′ terminal of the antisense strand, Z19 is the first nucleotide from the 3′ terminal of the sense strand:
the nucleotide difference between the nucleotide sequence II and the nucleotide sequence shown in SEQ ID NO:
242 comprises a difference at the site of Z20, and Z20 is selected from U, C or G;
vi) the nucleotide sequence I has the same length as and no more than three nucleotide differences from the nucleotide sequence shown in SEQ ID NO:
301;
and the nucleotide sequence II has the same length as and no more than three nucleotide differences from the nucleotide sequence shown in SEQ ID NO:
302:
(SEQ ID NO:
301)
5′-AACUUUCUAUUUCAAGAUZ21-3′;
(SEQ ID NO:
302)
5′-Z22AUCUUGAAAUAGAAAGUU-3′,
wherein, Z21 is U, and Z22 is A;
and the nucleotide sequence I comprises a nucleotide Z23 at a corresponding site to Z21, the nucleotide sequence II comprises a nucleotide Z24 at a corresponding site to Z22, and Z24 is the first nucleotide from the 5′ terminal of the antisense strand;
wherein “corresponding site” means being at the same site in the nucleotide sequence by counting from the same terminal of the nucleotide sequence, Z23 is the first nucleotide from the 3′ terminal of the sense strand:
the nucleotide difference between the nucleotide sequence II and the nucleotide sequence shown in SEQ ID NO:
302 comprises a difference at the site of Z24, and Z24 is selected from U, C or G;
R2 is a linear alkylene of 1-20 carbon atoms in length, wherein one or more carbon atoms are optionally replaced with any one or more selected from the group consisting of:
C(O), NH, O, S, CH═N, S(O)2, C2-C10 alkenylene, C2-C10 alkynylene, C6-C10 arylene, C3-C18 heterocyclylene, and C5-C10 heteroarylene;
and wherein R2 can optionally have any one or more substituents in the group consisting of:
C1-C10 alkyl, C6-C10 aryl, C5-C10 heteroaryl, C1-C10 haloalkyl, —OC1-C10 alkyl, —OC1-C10 alkylphenyl, —C1-C10 alkyl-OH, —OC1-C10 haloalkyl, —SC1-C10 alkyl, —SC1-C10 alkylphenyl, —C1-C10 alkyl-SH, —SC1-C10 haloalkyl, halo substituent, —OH, —SH, —NH2, —C1-C10 alkyl-NH2, —N(C1-C10 alkyl)(C1-C10 alkyl), —NH(C1-C10 alkyl), —N(C1-C10 alkyl)(C1-C10 alkylphenyl), —NH(C1-C10 alkylphenyl), cyano, nitro, —CO2H, —C(O)O(C1-C10 alkyl), —CON(C1-C10 alkyl)(C1-C10 alkyl), —CONH(C1-C10 alkyl), —CONH2, —NHC(O)(C1-C10 alkyl), —NHC(O)(phenyl), —N(C1-C10 alkyl)C(O)(C1-C10 alkyl), —N(C1-C10 alkyl)C(O)(phenyl), —C(O)C1-C10 alkyl, —C(O)C1-C10 alkylphenyl, —C(O)C1-C10 haloalkyl, —OC(O)C1-C10 alkyl, —SO2(C1-C10 alkyl), —SO2(phenyl), —SO2(C1-C10 haloalkyl), —SO2NH2, —SO2NH(C1-C10 alkyl), —SO2NH(phenyl), —NHSO2(C1-C10 alkyl), —NHSO2(phenyl), and —NHSO2(C1-C10 haloalkyl);
each L1 is a linear alkylene of 1-70 carbon atoms in length, wherein one or more carbon atoms are optionally replaced with any one or more selected from the group consisting of:
C(O), NH, O, S, CH═N, S(O)2, C2-C10 alkenylene, C2-C10 alkynylene, C6-C10 arylene, C3-C18 heterocyclylene, and C5-C10 heteroarylene;
and wherein L1 can optionally have any one or more substituents in the group consisting of:
C1-C10 alkyl, C6-C10 aryl, C5-C10 heteroaryl, C1-C10 haloalkyl, —OC1-C10 alkyl, —OC1-C10 alkylphenyl, —C1-C10 alkyl-OH, —OC1-C10 haloalkyl, —SC1-C10 alkyl, —SC1-C10 alkylphenyl, —C1-C10 alkyl-SH, —SC1-C10 haloalkyl, halo substituent, —OH, —SH, —NH2, —C1-C10 alkyl-NH2, —N(C1-C10 alkyl)(C1-C10 alkyl), —NH(C1-C10 alkyl), —N(C1-C10 alkyl)(C1-C10 alkylphenyl), —NH(C1-C10 alkylphenyl), cyano, nitro, —CO2H, —C(O)O(C1-C10 alkyl), —CON(C1-C10 alkyl)(C1-C10 alkyl), —CONH(C1-C10 alkyl), —CONH2, —NHC(O)(C1-C10 alkyl), —NHC(O)(phenyl), —N(C1-C10 alkyl)C(O)(C1-C10 alkyl), —N(C1-C10 alkyl)C(O)(phenyl), —C(O)C1-C10 alkyl, —C(O)C1-C10 alkylphenyl, —C(O)C1-C10 haloalkyl, —OC(O)C1-C10 alkyl, —SO2(C1-C10 alkyl), —SO2(phenyl), —SO2(C1-C10 haloalkyl), —SO2NH2, —SO2NH(C1-C10 alkyl), —SO2NH(phenyl), —NHSO2(C1-C10 alkyl), —NHSO2(phenyl), and —NHSO2(C1-C10 haloalkyl);
represents the site where a group is covalently linked;
and
M1 represents a targeting group.

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Nucleic acid, pharmaceutical composition, conjugate, preparation method, and use

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Nucleic acid, pharmaceutical composition, conjugate, preparation method, and use

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Abstract

Claims8