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

An siRNA conjugate, wherein the siRNA conjugate has a structure as shown by Formula (308):

• wherein,
• n1 is an integer of 1-2, and n3 is an integer of 0-1, and n1+n3=2-3:
• m1, m2, and m3 independently of one another are an integer of 2-10;
• R10, R11, R12, R13, R14, and R15 independently of one another are 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;
• and Nu is siRNA,
• wherein the siRNA comprises a sense strand and an antisense strand, each nucleotide in the siRNA being 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;
• wherein the sense strand and the antisense strand have the same or different length, wherein the sense strand has a length of 19 to 23 nucleotides, and the antisense strand has a length of 19 to 26 nucleotides;
• and
• wherein the nucleotide sequence I and the nucleotide sequence II are selected from the sequences as shown in any of i) to ix):
• i) the nucleotide sequence I has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 1 with no more than 3 nucleotide differences therebetween, and the nucleotide sequence II has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 2 with no more than 3 nucleotide differences therebetween:
• (SEQ ID NO:
• 1)
• 5′-GGGUAUUCUUUCAAGCAAZ1-3′;
• (SEQ ID NO:
• 2)
• 5′-Z2UUGCUUGAAAGAAUACCC-3′,
• wherein, Z1 is U and Z2 is A, 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 has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 61 with no more than 3 nucleotide differences therebetween, and the nucleotide sequence II has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 62 with no more than 3 nucleotide differences therebetween:
• (SEQ ID NO:
• 61)
• 5′-GGCAUAAACUAUAACAGCZ5-3′;
• (SEQ ID NO:
• 62)
• 5′-Z6GCUGUUAUAGUUUAUGCC-3′,
• wherein, Z5 is U and Z6 is A, 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 has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 121 with no more than 3 nucleotide differences therebetween, and the nucleotide sequence II has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 122 with no more than 3 nucleotide differences therebetween:
• (SEQ ID NO:
• 121)
• 5′-GCUCAAGAAUGCCAAGAAZ9-3′;
• (SEQ ID NO:
• 122)
• 5′-Z10UUCUUGGCAUUCUUGAGC-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 has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 181 with no more than 3 nucleotide differences therebetween, and the nucleotide sequence II has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 182 with no more than 3 nucleotide differences therebetween:
• (SEQ ID NO:
• 181)
• 5′-GCAACAAAGACAUUUAUGZ13-3′;
• (SEQ ID NO:
• 182)
• 5′-Z14CAUAAAUGUCUUUGUUGC-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 has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 241 with no more than 3 nucleotide differences therebetween, and the nucleotide sequence II has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 242 with no more than 3 nucleotide differences therebetween:
• (SEQ ID NO:
• 241)
• 5′-GAAUCUCAAAGAAAUCUUZ17-3′;
• (SEQ ID NO:
• 242)
• 5′-Z18AAGAUUUCUUUGAGAUUC-3′,
• wherein, Z1 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;
• vi) the nucleotide sequence I has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 301 with no more than 3 nucleotide differences therebetween, and the nucleotide sequence II has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 302 with no more than 3 nucleotide differences therebetween:
• (SEQ ID NO:
• 301)
• 5′-GUACGUGGACUGGAUUCUZ21-3′;
• (SEQ ID NO:
• 302)
• 5′-Z22AGAAUCCAGUCCACGUAC-3′,
• wherein, Z21 is G and Z22 is C, 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;
• vii) the nucleotide sequence I has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 361 with no more than 3 nucleotide differences therebetween, and the nucleotide sequence II has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 362 with no more than 3 nucleotide differences therebetween:
• (SEQ ID NO:
• 361)
• 5′-AUUUCUGGGUAUUCUUUCZ25-3′;
• (SEQ ID NO:
• 362)
• 5′-Z26GAAAGAAUACCCAGAAAU-3′,
• wherein, Z25 is A and Z26 is U, and
• the nucleotide sequence I comprises a nucleotide Z27 at the position corresponding to Z25;
• the nucleotide sequence II comprises a nucleotide Z28 at the position corresponding to Z26, wherein Z28 is the first nucleotide at 5′ terminal of the antisense strand;
• viii) the nucleotide sequence I has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 421 with no more than 3 nucleotide differences therebetween, and the nucleotide sequence II has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 422 with no more than 3 nucleotide differences therebetween:
• (SEQ ID NO:
• 421)
• 5′-CAUGAAGGGCAUAAACUAZ29-3′;
• (SEQ ID NO:
• 422)
• 5′-Z30UAGUUUAUGCCCUUCAUG-3′,
• wherein, Z29 is U and Z30 is A, and
• the nucleotide sequence I comprises a nucleotide Z31 at the position corresponding to Z29;
• the nucleotide sequence II comprises a nucleotide Z32 at the position corresponding to Z30, wherein Z32 is the first nucleotide at 5′ terminal of the antisense strand;
• ix) the nucleotide sequence I has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 481 with no more than 3 nucleotide differences therebetween, and the nucleotide sequence II has the same length as the nucleotide sequence as shown by SEQ ID NO:
• 482 with no more than 3 nucleotide differences therebetween:
• (SEQ ID NO:
• 481)
• 5′-GGAUUCUGGAGAAAACUCZ33-3′;
• (SEQ ID NO:
• 482)
• 5′-Z34GAGUUUUCUCCAGAAUCC-3′,
• wherein, Z33 is A and Z34 is U, and
• the nucleotide sequence I comprises a nucleotide Z35 at the position corresponding to Z33;
• the nucleotide sequence II comprises a nucleotide Z36 at the position corresponding to Z34, wherein Z36 is the first nucleotide at 5′ terminal of the antisense strand_;
• “corresponding position” refers to a same position in the nucleotide sequence by 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 alkylOH, OC1-C10 haloalkyl, SC1-C10 alkyl, SC1-C10 alkylphenyl, —C1-C10 alkylSH, SC1-C10 haloalkyl, halo, OH, —SH, NH2, C1-C10 alkylNH2, 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 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 alkylOH, OC1-C10 haloalkyl, SC1-C10 alkyl, SC1-C10 alkylphenyl, —C1-C10 alkylSH, SC1-C10 haloalkyl, halo, OH, —SH, NH2, C1-C10 alkylNH2, 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 the group is covalently linked;
• M1 represents a targeting group, wherein each of the targeting groups is independently a ligand that has affinity to asialoglycoprotein receptors on a surface of mammalian hepatocytes.