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

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

• wherein
• n1 is an integer of 1-2, n3 is an integer of 0-1, and n1+n3=2-3;
• each m1, m2, or m3 independently of one another is an integer of 2-10;
• R10, R11, R12, R13, R14, or R15 independently of one another is H, or selected from the group consisting of C1-C10 alkyl, C1-C10 haloalkyl, and C1-C10 alkoxy;
• R3 is a group having a structure as shown by Formula (A59):
• wherein,
• E1 is OH, SH or BH2;
• Nu is an siRNA;
• the siRNA comprises a sense strand and an antisense strand;
• 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;
• the nucleotide sequence I and the nucleotide sequence II are the sequences selected from one of the following groups i)-vi):
• i) the nucleotide sequence I and the nucleotide sequence as shown by SEQ ID NO:
• 1 have an equal length and no more than 3 nucleotide differences, and the nucleotide sequence II and the nucleotide sequence as shown by SEQ ID NO:
• 2 have an equal length and no more than 3 nucleotide differences:
• (SEQ ID NO:
• 1)
• 5′-AAGCAAGCAGACAUUUAUZ1-3′;
• (SEQ ID NO:
• 2)
• 5′-Z2AUAAAUGUCUGCUUGCUU-3′,
• wherein, Z1 is C and Z2 is G, and
• the nucleotide sequence I comprises a nucleotide Z3 at the position corresponding to Z1;
• the nucleotide sequence II comprises a nucleotide Z4 at the position corresponding to Z2, wherein Z4 is the first nucleotide at 5′ terminal of the antisense strand;
• ii) the nucleotide sequence I and the nucleotide sequence as shown by SEQ ID NO:
• 61 have an equal length and no more than 3 nucleotide differences, and the nucleotide sequence II and the nucleotide sequence as shown by SEQ ID NO:
• 62 have an equal length and no more than 3 nucleotide differences:
• (SEQ ID NO:
• 61)
• 5′-UUUGUAGCAUUUUUAUUAZ5-3′;
• (SEQ ID NO:
• 62)
• 5′-Z6UAAUAAAAAUGCUACAAA-3′,
• wherein, Z5 is A and Zb is U, and
• the nucleotide sequence I comprises a nucleotide Z7 at the position corresponding to Z5;
• the nucleotide sequence II comprises a nucleotide Z8 at the position corresponding to Z6, wherein Z8 is the first nucleotide at 5′ terminal of the antisense strand;
• iii) the nucleotide sequence I and the nucleotide sequence as shown by SEQ ID NO:
• 121 have an equal length and no more than 3 nucleotide differences, and the nucleotide sequence II and the nucleotide sequence as shown by SEQ ID NO:
• 122 have an equal length and no more than 3 nucleotide differences:
• (SEQ ID NO:
• 121)
• 5′-GCCUGGAGUUUAUUCGGAZ9-3′;
• (SEQ ID NO:
• 122)
• 5′-Z10UCCGAAUAAACUCCAGGC-3′,
• wherein, Z9 is A and Z10 is U, and
• the nucleotide sequence I comprises a nucleotide Z11 at the position corresponding to Z9;
• the nucleotide sequence II comprises a nucleotide Z12 at the position corresponding to Z10, wherein Z12 is the first nucleotide at 5′ terminal of the antisense strand;
• iv) the nucleotide sequence I and the nucleotide sequence as shown by SEQ ID NO:
• 181 have an equal length and no more than 3 nucleotide differences, and the nucleotide sequence II and the nucleotide sequence as shown by SEQ ID NO:
• 182 have an equal length and no more than 3 nucleotide differences:
• (SEQ ID NO:
• 181)
• 5′-CUGUUUUGCUUUUGUAACZ13-3′;
• (SEQ ID NO:
• 182)
• 5′-Z14GUUACAAAAGCAAAACAG-3′,
• wherein, Z13 is U and Z14 is A, and
• the nucleotide sequence I comprises a nucleotide Z15 at the position corresponding to Z13;
• the nucleotide sequence II comprises a nucleotide Z16 at the position corresponding to Z14, wherein Z16 is the first nucleotide at 5′ terminal of the antisense strand;
• v) the nucleotide sequence I and the nucleotide sequence as shown by SEQ ID NO:
• 241 have an equal length and no more than 3 nucleotide differences, and the nucleotide sequence II and the nucleotide sequence as shown by SEQ ID NO:
• 242 have an equal length and no more than 3 nucleotide differences:
• (SEQ ID NO:
• 241)
• 5′-GGUUUUGUAGCAUUUUUAZ17-3′;
• (SEQ ID NO:
• 242)
• 5′-Z18UAAAAAUGCUACAAAACC-3′,
• wherein, Z17 is U and Z18 is A, and
• the nucleotide sequence I comprises a nucleotide Z19 at the position corresponding to Z17;
• the nucleotide sequence II comprises a nucleotide Z20 at the position corresponding to Z18, wherein Z20 is the first nucleotide at 5′ terminal of the antisense strand;
• and
• vi) the nucleotide sequence I and the nucleotide sequence as shown by SEQ ID NO:
• 301 have an equal length and no more than 3 nucleotide differences, and the nucleotide sequence II and the nucleotide sequence as shown by SEQ ID NO:
• 302 have an equal length and no more than 3 nucleotide differences:
• (SEQ ID NO:
• 301)
• 5′-GUGACUUUUUAAAAUAAAZ21-3′;
• (SEQ ID NO:
• 302)
• 5′-Z22UUUAUUUUAAAAAGUCAC-3′,
• wherein, Z21 is A and Z22 is U, and
• the nucleotide sequence I comprises a nucleotide Z23 at the position corresponding to Z21;
• the nucleotide sequence II comprises a nucleotide Z24 at the position corresponding to Z22, wherein Z24 is the first nucleotide at 5′ terminal of the antisense strand;
• “corresponding position” refers to a same position in a nucleotide sequence when counting from a same terminal of the nucleotide sequence;
• R2 is a linear alkylene of 1 to 20 carbon atoms in length, wherein one or more carbon atoms are optionally replaced with any one or more groups 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 optionally has any one or more substituents selected from 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, —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 independently is a linear alkylene of 1 to 70 carbon atoms in length, wherein one or more carbon atoms are optionally replaced with any one or more groups 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 optionally has any one or more substituents selected from 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, —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.