JPWO2020233655A5

JPWO2020233655A5 -

Info

Publication number: JPWO2020233655A5
Application number: JP2021569135A
Authority: JP
Publication date: 2023-05-31

Claims

An siRNA complex having a structure represented by formula (308),

During the ceremony,
n1 is an integer selected from 1 to 3, n3 is an integer selected from 0 to 4, and each m1, m2 or m3 is independently an integer selected from 2 to 10 , R ₁₀ , R ₁₁ , R ₁₂ , R ₁₃ , R ₁₄ or R ₁₅ are each independently H or a C ₁ -C ₁₀ alkyl group, a C ₁ -C ₁₀ halogenated alkyl group and a C ₁ - is selected from the group consisting of _C10 alkoxy groups,
R ₃ is a group of the structure shown in formula A59,

wherein E ₁ is OH, SH or BH ₂ ,
Nu is siRNA, said siRNA comprises a sense strand and an antisense strand, each nucleotide of said siRNA is independently a modified or unmodified nucleotide, said sense strand comprises nucleotide sequence I, said antisense strand The strand comprises a nucleotide sequence II, wherein said nucleotide sequence I and said nucleotide sequence II form a double-stranded region, at least in part, in reverse complementarity, said nucleotide sequence I and said nucleotide sequence II having the following one selected from the sequences shown in i) to vi),
i) said nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 1 are equal in length and differ by no more than 3 nucleotides, and said nucleotide sequence II and the nucleotide sequence shown in SEQ ID NO: 2 are are equal and have no more than 3 nucleotide differences,
5′-AAGCAAGCAGACAUUUAUZ _1-3 ′ (SEQ ID NO: 1),
5′-Z ₂ AUAAAUGUCUGCUUGCUU-3′ (SEQ ID NO: 2)
with the proviso that Z ₁ is C and Z ₂ is G, said nucleotide sequence I includes a nucleotide Z ₃ corresponding to position Z ₁ and said nucleotide sequence II includes a nucleotide corresponding to position Z ₂ Z ₄ , said Z ₄ being the first nucleotide at the 5' end of said antisense strand;
ii) said nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 61 are equal in length and differ by no more than 3 nucleotides, and said nucleotide sequence II and the nucleotide sequence shown in SEQ ID NO: 62 are are equal and have no more than 3 nucleotide differences,
5′-UUUGUAGCAUUUUUUUAZ _5-3 ′ (SEQ ID NO: 61),
5'- _{Z6UAAUAAAAAAUGCUACAAA} -3' (SEQ ID NO: 62)
with the proviso that Z ₅ is A and Z ₆ is U, said nucleotide sequence I includes a nucleotide Z ₇ corresponding to position Z ₅ , and said nucleotide sequence II includes a nucleotide corresponding to position Z ₆ Z ₈ , said Z ₈ being the first nucleotide at the 5' end of said antisense strand;
iii) said nucleotide sequence I and the nucleotide sequence set forth in SEQ ID NO: 121 are equal in length and differ by no more than 3 nucleotides, and said nucleotide sequence II and the nucleotide sequence set forth in SEQ ID NO: 122 are equal in length to are equal and have no more than 3 nucleotide differences,
5′-GCCUGGAGUUUAUUCGGAZ _9-3 ′ (SEQ ID NO: 121),
5′-Z ₁₀ UCCGAAUAAACUCCAGGC-3′ (SEQ ID NO: 122)
with the proviso that Z ₉ is A and Z ₁₀ is U, said nucleotide sequence I includes a nucleotide Z ₁₁ corresponding to position Z ₉ , and said nucleotide sequence II includes a nucleotide corresponding to Z ₁₀ at position Z ₁₂ , said Z ₁₂ being the first nucleotide at the 5' end of said antisense strand;
iv) said nucleotide sequence I and the nucleotide sequence set forth in SEQ ID NO: 181 are equal in length and differ by no more than 3 nucleotides, and said nucleotide sequence II and the nucleotide sequence set forth in SEQ ID NO: 182 are equal in length to are equal and have no more than 3 nucleotide differences,
5′-CUGUUUUGCUUUUGUAACZ ₁₃ -3′ (SEQ ID NO: 181),
5′-Z ₁₄ GUUACAAAAGCAAAACAG-3′ (SEQ ID NO: 182)
with the proviso that Z ₁₃ is U and Z ₁₄ is A, said nucleotide sequence I includes a nucleotide Z ₁₅ corresponding to Z ₁₃ at position, and said nucleotide sequence II includes a nucleotide corresponding to Z ₁₄ at position Z ₁₆ , said Z ₁₆ being the first nucleotide at the 5' end of said antisense strand;
v) said nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 241 are equal in length and differ by no more than 3 nucleotides, and said nucleotide sequence II and the nucleotide sequence shown in SEQ ID NO: 242 are are equal and have no more than 3 nucleotide differences,
5′-GGUUUUGUAGCAUUUUUAZ ₁₇ -3′ (SEQ ID NO: 241),
5′-Z ₁₈ UAAAAAAUGCUACAAAACC-3′ (SEQ ID NO: 242)
with the proviso that Z ₁₇ is U and Z ₁₈ is A, said nucleotide sequence I includes a nucleotide Z ₁₉ corresponding to position Z ₁₇ , and said nucleotide sequence II includes a nucleotide corresponding to Z ₁₈ at position Z ₂₀ , said Z ₂₀ being the first nucleotide at the 5' end of said antisense strand;
vi) said nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 301 are equal in length and differ by no more than 3 nucleotides, and said nucleotide sequence II and the nucleotide sequence shown in SEQ ID NO: 302 are are equal and have no more than 3 nucleotide differences,
5′-GUGACUUUUUAAAAUAAAZ _21-3 ′ (SEQ ID NO: 301),
5′-Z ₂₂ UUUAUUUUAAAAGUCAC-3′ (SEQ ID NO: 302)
with the proviso that Z ₂₁ is A and Z ₂₂ is U, said nucleotide sequence I includes a nucleotide Z ₂₃ corresponding to position Z ₂₁ and said nucleotide sequence II includes a nucleotide corresponding to position Z ₂₂ Z ₂₄ , said Z ₂₄ being the first nucleotide at the 5' end of said antisense strand;
R ₂ is a linear alkylene group of 1 to 20 carbon atoms in length, in which one or more carbon atoms are C(O), NH, O, S, CH═N, S(O) ₂ , C ₂ -C ₁₀ alkenylene groups, C ₂ -C ₁₀ alkynylene groups, C ₆ -C ₁₀ arylene groups, C ₃ -C ₁₈ heterocyclylene groups and C ₅ -C ₁₀ heteroarylene groups optionally substituted with one or more, R ₂ is a C ₁ -C ₁₀ alkyl group, a C ₆ -C ₁₀ aryl group, a C ₅ -C ₁₀ heteroaryl group, a C ₁ -C ₁₀ halogenated alkyl group, —OC ₁ -C ₁₀ alkyl group, -OC ₁ -C ₁₀ alkylphenyl group, -C ₁ -C ₁₀ alkyl-OH, -OC ₁ -C ₁₀ halogenated alkyl group, -SC ₁ -C ₁₀ alkyl group, -SC ₁ -C ₁₀ alkylphenyl group, -C ₁ -C ₁₀ alkyl-SH, -SC ₁ -C ₁₀ halogenated alkyl group, halogen substituent, -OH, -SH, -NH ₂ , -C ₁ -C ₁₀ alkyl- NH ₂ , —N(C ₁ -C ₁₀ alkyl group) (C ₁ -C ₁₀ alkyl group), —NH (C ₁ -C ₁₀ alkyl group), —N(C ₁ -C ₁₀ alkyl group) (C ₁ —C ₁₀ alkylphenyl group), —NH (C ₁ -C ₁₀ alkylphenyl group), cyano group, nitro group, —CO ₂ H, —C(O)O (C ₁ -C ₁₀ alkyl group), —CON (C ₁ -C ₁₀ alkyl group) (C ₁ -C ₁₀ alkyl group), -CONH (C ₁ -C ₁₀ alkyl group), -CONH ₂ , -NHC(O) (C ₁ -C ₁₀ alkyl group), —NHC(O) (phenyl group), —N (C ₁ -C ₁₀ alkyl group) C(O) (C ₁ -C ₁₀ alkyl group), —N (C ₁ -C ₁₀ alkyl group) C(O) (phenyl group), -C(O)C1 _- _C10 alkyl group, -C(O) _C1 - _C10 alkylphenyl group, -C(O)C1 _- _C10 haloalkyl group, -OC(O) C ₁ -C ₁₀ alkyl group, -SO ₂ (C ₁ -C ₁₀ alkyl group), -SO ₂ (phenyl group), -SO ₂ (C ₁ -C ₁₀ halogenated alkyl group), -SO ₂ NH ₂ , —SO ₂ NH (C ₁ -C ₁₀ alkyl group), —SO ₂ NH (phenyl group), —NHSO ₂ (C ₁ -C ₁₀ alkyl group), —NHSO ₂ (phenyl group) and —NHSO ₂ (C ₁ —C ₁₀ halogenated alkyl groups), optionally having any one or more substituents,
Each L ₁ is independently a linear alkylene group of 1 to 70 carbon atoms in length, wherein one or more carbon atoms are C(O), NH, O, S, CH═N, S(O) ₂ , consisting of a C ₂ -C ₁₀ alkenylene group, a C ₂ -C ₁₀ alkynylene group, a C ₆ -C ₁₀ arylene group, a C ₃ -C ₁₈ heterocyclylene group and a C ₅ -C ₁₀ heteroarylene group; optionally substituted with one or more selected from the group, L ₁ is a C ₁ -C ₁₀ alkyl group, a C ₆ -C ₁₀ aryl group, a C ₅ -C ₁₀ heteroaryl group, a C ₁ -C ₁₀ halogenated alkyl group, -OC ₁ -C ₁₀ alkyl group, -OC ₁ -C ₁₀ alkylphenyl group, -C ₁ -C ₁₀ alkyl-OH, -OC ₁ -C ₁₀ halogenated alkyl group, -SC ₁ -C ₁₀ alkyl group, -SC ₁ -C ₁₀ alkylphenyl group, -C ₁ -C ₁₀ alkyl-SH, -SC ₁ -C ₁₀ halogenated alkyl group, halogen substituent, -OH, -SH, -NH ₂ , -C ₁ —C ₁₀ alkyl-NH ₂ , —N(C ₁ -C ₁₀ alkyl group) (C ₁ -C ₁₀ alkyl group), —NH (C ₁ -C ₁₀ alkyl group), —N(C ₁ -C ₁₀ alkyl group) group) (C ₁ -C ₁₀ alkylphenyl group), —NH (C ₁ -C ₁₀ alkylphenyl group), cyano group, nitro group, —CO ₂ H, —C(O)O(C ₁ -C ₁₀ alkyl group), -CON (C ₁ -C ₁₀ alkyl group) (C ₁ -C ₁₀ alkyl group), -CONH (C ₁ -C ₁₀ alkyl group), -CONH ₂ , -NHC(O) (C ₁ -C ₁₀ alkyl group), —NHC(O) (phenyl group), —N (C ₁ -C ₁₀ alkyl group) C(O) (C ₁ -C ₁₀ alkyl group), —N (C ₁ -C ₁₀ alkyl group ) C(O) (phenyl group), —C(O) C ₁ -C ₁₀ alkyl group, —C(O) C ₁ -C ₁₀ alkylphenyl group, —C(O) C ₁ -C ₁₀ haloalkyl group, —OC(O) C ₁ -C ₁₀ alkyl group, —SO ₂ (C ₁ -C ₁₀ alkyl group), —SO ₂ (phenyl group), —SO ₂ (C ₁ -C ₁₀ halogenated alkyl group), — SO ₂ NH ₂ , —SO ₂ NH (C ₁ -C ₁₀ alkyl group), —SO ₂ NH (phenyl group), —NHSO ₂ (C ₁ -C ₁₀ alkyl group), —NHSO ₂ (phenyl group) and — optionally having any one or more substituents from the group consisting of NHSO ₂ (C ₁ -C ₁₀ halogenated alkyl groups);

siRNA conjugates, where represents the site to which the group is covalently attached and _M1 represents the targeting group.

each L ₁ is independently selected from the group consisting of (A1) to (A26) groups and any combination thereof;

wherein each j1 is independently an integer from 1 to 20 and each j2 is independently an integer from 1 to 20;
each R′ is independently a C ₁ -C ₁₀ alkyl group;
each Ra is independently selected from the group consisting of groups represented by formulas (A27) to (A45) or any combination thereof;

each Rb is independently a C ₁ -C ₁₀ alkyl group;
Alternatively, L ₁ is one or a combination of bonds selected from A1, A4, A5, A6, A8, A10, A11, A13;
Alternatively, L ₁ is a combination of at least two bonds selected from A1, A4, A8, A10 and A11;
or L ₁ is a combination of at least two bonds selected from A1, A8, A10;
or L ₁ is 3 to 25 atoms in length;
or L ₁ is 4 to 15 atoms in length;

2. The siRNA complex of claim 1, wherein represents the site to which the group is covalently attached.

n1 is an integer of 1 to 2, n3 is an integer of 0 to 1, and n1+n3=2 to 3, or
2. The siRNA complex of claim 1 , wherein each m1, m2 or m3 is independently an integer from 2 to 5 and/or m1=m2=m3.

wherein each said targeting group is independently a ligand having affinity for asialoglycoprotein receptors on the surface of mammalian hepatocytes;
or each said targeting group is independently an asialoglycoprotein or a sugar;
Alternatively, each said targeting group is D-mannopyranose, L-mannopyranose, D-arabinose, D-xylofuranose, L-xylofuranose, D-glucose, L-glucose, D-galactose, L-galactose, α-D-mannofuranose, β-D-mannofuranose, α-D-mannopyranose, β-D-mannopyranose, α-D-glucopyranose, β-D-glucopyranose, α-D- Glucofuranose, β-D-glucofuranose, α-D-fructofuranose, α-D-fructopyranose, α-D-galactopyranose, β-D-galactopyranose, α-D-galactofuranose, β-D - galactofuranose, glucosamine, sialic acid, galactosamine, N-acetylgalactosamine, N-trifluoroacetylgalactosamine, N-propionylgalactosamine, Nn-butyrylgalactosamine, N-isobutyrylgalactosamine, 2-amino-3-O -[(R)-1-carboxyethyl]-2-deoxy-β-D-glucopyranose, 2-deoxy-2-methylamino-L-glucopyranose, 4,6-dideoxy-4-formamide-2,3 -di-O-methyl-D-mannopyranose, 2-deoxy-2-sulfoamino-D-glucopyranose, N-glycolyl-α-neuraminic acid, 5-thio-β-D-glucopyranose, methyl 2,3 , 4-tris-O-acetyl-1-thio-6-O-trityl-α-D-glucopyranoside, 4-thio-β-D-galactopyranose, ethyl 3,4,6,7-tetra-O-acetyl -2-deoxy-1,5-dithio-α-D-glucoheptopyranoside, 2,5-anhydro-D-aronitrile, ribose, D-ribose, D-4-thioribose, L-ribose, L-4 - one independently selected from thioribose;
Alternatively, the siRNA complex of claim 1 , wherein at least one or each of said targeting groups is galactose or N-acetylgalactosamine.

Equations (403), (404), (405), (406), (407), (408), (409), (410), (411), (412), (413), (414), ( 415), (416), (417), (418), (419), (420), (421) or (422) .

the P atom in formula A59 is attached to the end of the sense or antisense strand of the siRNA, said end referring to the 4 nucleotides preceding one end of said sense or antisense strand;
Alternatively, the P atom in formula A59 is attached to the end of the sense or antisense strand of the siRNA, or the P atom in formula A59 is attached to the 3' end of the sense strand of the siRNA,
Alternatively, the siRNA complex of claim 1 , wherein the P atom in formula A59 is attached to the 2', 3' or 5' position of a nucleotide in said siRNA by a phosphodiester bond.

the sense strand further comprises nucleotide sequence III, the antisense strand further comprises nucleotide sequence IV, and the length of nucleotide sequence III and nucleotide sequence IV are each independently 1-4 nucleotides, and said nucleotide sequence III is attached to the 5′ end of nucleotide sequence I and nucleotide sequence IV is attached to the 3′ end of nucleotide sequence II, wherein said nucleotide sequence III and said nucleotide sequence IV are equal in length and substantially reverse complementary or fully reverse complementary, said substantially reverse complementary refers to the presence of one or less base mismatches between the two nucleotide sequences, and fully reverse complementary refers to the two nucleotide sequences 2. The siRNA complex of claim 1 , referring to no mismatches between.

i) said nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 3, said nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO: 4, and said nucleotide sequences III and IV each have a length of 1 is a nucleotide, and the base of the nucleotide sequence III is C, or the nucleotide sequences III and IV are both 2 nucleotides in length, and from the 5' end to the 3' end, The nucleotide sequence is CC, or the nucleotide sequences III and IV are both 3 nucleotides in length, and the nucleotide sequence of the nucleotide sequence III is CCC from the 5' end to the 3' end, or , the nucleotide sequences III and IV are both 4 nucleotides in length, and the base sequence of the nucleotide sequence III is ACCC from the 5' end to the 3' end, or
ii) said nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 63, said nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO: 64, and said nucleotide sequences III and IV are both 1 in length. is a nucleotide, and the base of the nucleotide sequence III is U, or the nucleotide sequences III and IV are both 2 nucleotides in length, and from the 5' end to the 3' end of the nucleotide sequence III The nucleotide sequence is GU, or the nucleotide sequences III and IV are both 3 nucleotides in length, and the nucleotide sequence of the nucleotide sequence III is GGU from the 5' end to the 3' end, or , the nucleotide sequences III and IV are both 4 nucleotides in length, and the base sequence of the nucleotide sequence III is GGGU from the 5' end to the 3' end, or
iii) said nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 123, said nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO: 124, and said nucleotide sequences III and IV are both 1 in length. is a nucleotide, and the base of the nucleotide sequence III is G, or the nucleotide sequences III and IV are both 2 nucleotides in length, and from the 5' end to the 3' end of the nucleotide sequence III The nucleotide sequence is AG, or the nucleotide sequences III and IV are both 3 nucleotides in length, and the nucleotide sequence of the nucleotide sequence III is UAG from the 5' end to the 3' end, or , the nucleotide sequences III and IV are both 4 nucleotides in length, and the base sequence of the nucleotide sequence III is AUAG from the 5' end to the 3' end, or
iv) said nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 183, said nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO: 184, and said nucleotide sequences III and IV are both 1 in length. is a nucleotide, and the base of the nucleotide sequence III is C, or the nucleotide sequences III and IV are both 2 nucleotides in length, and from the 5' end to the 3' end, The nucleotide sequence is AC, or the nucleotide sequences III and IV are both 3 nucleotides in length, and the nucleotide sequence of the nucleotide sequence III is GAC from the 5' end to the 3' end, or , the nucleotide sequences III and IV are both 4 nucleotides in length, and the base sequence of the nucleotide sequence III is AGAC from the 5' end to the 3' end, or
v) said nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 243, said nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO: 244, and said nucleotide sequences III and IV are both 1 in length is a nucleotide, and the base of the nucleotide sequence III is G, or the nucleotide sequences III and IV are both 2 nucleotides in length, and from the 5' end to the 3' end of the nucleotide sequence III The nucleotide sequence is UG, or the nucleotide sequences III and IV are both 3 nucleotides in length, and the nucleotide sequence of the nucleotide sequence III is CUG from the 5' end to the 3' end, or , the nucleotide sequences III and IV are both 4 nucleotides in length, and the base sequence of the nucleotide sequence III is UCUG from the 5' end to the 3' end, or
vi) said nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 303, said nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO: 304, and said nucleotide sequences III and IV are both 1 in length is a nucleotide, and the base of the nucleotide sequence III is G, or the nucleotide sequences III and IV are both 2 nucleotides in length, and from the 5' end to the 3' end of the nucleotide sequence III The base sequence is UG, or the nucleotide sequences III and IV are both 3 nucleotides in length, and the base sequence of the nucleotide sequence III is AUG from the 5' end to the 3' end, or 8. The siRNA complex according to claim 7 , wherein each of said nucleotide sequences III and IV has a length of 4 nucleotides, and the base sequence of nucleotide sequence III is UAUG from the 5' end to the 3' end. .

The antisense strand further comprises a nucleotide sequence V, wherein the nucleotide sequence V is 1-3 nucleotides in length and is attached to the 3' end of the antisense strand to form a 3' overhanging end of the antisense strand. Alternatively, said nucleotide sequence V is 2 nucleotides in length, or said nucleotide sequence V is two consecutive thymine deoxyribonucleotides or two consecutive uracil ribonucleotides, or said 2. The siRNA complex of claim 1 , wherein nucleotide sequence V is complementary to the nucleotide at the corresponding position in the target mRNA.

The sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:5 and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:6, or the sense strand of the siRNA is shown in SEQ ID NO:7. comprising a nucleotide sequence, wherein the antisense strand comprises the nucleotide sequence set forth in SEQ ID NO:8;
Alternatively, the sense strand of the siRNA comprises the nucleotide sequence set forth in SEQ ID NO: 65, and the antisense strand comprises the nucleotide sequence set forth in SEQ ID NO: 66, or the sense strand of the siRNA contains the nucleotide sequence set forth in SEQ ID NO: 67. comprising the nucleotide sequence shown, wherein the antisense strand comprises the nucleotide sequence shown in SEQ ID NO: 68;
Alternatively, the siRNA sense strand comprises the nucleotide sequence set forth in SEQ ID NO: 125, and the siRNA antisense strand comprises the nucleotide sequence set forth in SEQ ID NO: 126, or the siRNA sense strand comprises the nucleotide sequence set forth in SEQ ID NO: 127, wherein the antisense strand of the siRNA comprises the nucleotide sequence set forth in SEQ ID NO: 128;
Alternatively, the siRNA sense strand comprises the nucleotide sequence set forth in SEQ ID NO: 185, and the antisense strand comprises the nucleotide sequence set forth in SEQ ID NO: 186, or the siRNA sense strand comprises the nucleotide sequence set forth in SEQ ID NO: 187. comprising the nucleotide sequence shown, wherein said antisense strand comprises the nucleotide sequence shown in SEQ ID NO: 188;
Alternatively, the siRNA sense strand comprises the nucleotide sequence set forth in SEQ ID NO: 245, and the siRNA antisense strand comprises the nucleotide sequence set forth in SEQ ID NO: 246, or the siRNA sense strand comprises the nucleotide sequence set forth in SEQ ID NO: 247, wherein the antisense strand of said siRNA comprises the nucleotide sequence set forth in SEQ ID NO: 248;
Alternatively, the siRNA sense strand comprises the nucleotide sequence set forth in SEQ ID NO: 305, and the siRNA antisense strand comprises the nucleotide sequence set forth in SEQ ID NO: 306, or the siRNA sense strand comprises the nucleotide sequence set forth in SEQ ID NO: 307. The siRNA complex of claim 1 , comprising the nucleotide sequence set forth in SEQ ID NO:308, wherein the antisense strand of said siRNA comprises the nucleotide sequence set forth in SEQ ID NO:308.

2. The siRNA complex of claim 1 , wherein the siRNA has the nucleotide sequence set forth in siPCSKa1, siPCSKa2, siPCSKb1, siPCSKb2, siPCSKc1, siPCSKc2, siPCSKd1, siPCSKd2, siPCSKe1, siPCSKe2, siPCSKf1 or siPCSKf2.

each nucleotide in the sense strand and the antisense strand is independently a fluoro-modified nucleotide or a non-fluoro-modified nucleotide;
Alternatively, said fluoro-modified nucleotides are located in nucleotide sequence I and nucleotide sequence II, and from the 5' end to the 3' end, at least nucleotides at positions 7, 8 and 9 of said nucleotide sequence I are fluoro-modified nucleotides. wherein, from the 5' end to the 3' end, at least nucleotides at positions 2, 6, 14, and 16 of said nucleotide sequence II are fluoro-modified nucleotides;
Alternatively, from the 5' end to the 3' end, in the sense strand, nucleotides at positions 7, 8, 9 or 5, 7, 8, 9 of the nucleotide sequence I are fluoro-modified nucleotides; 2, 6, 14, 16 or 2, 6 of said nucleotide sequence II in said antisense strand, from the 5' end to the 3' end, wherein the nucleotides at the remaining positions in the strand are non-fluoro modified nucleotides; , 8, 9, 14, 16 are fluoro-modified nucleotides, and the remaining nucleotides in the antisense strand are non-fluoro-modified nucleotides .

each non-fluoro-modified nucleotide is one independently selected from nucleotides or nucleotide analogs in which the hydroxy group at the 2' position of the ribose group of the nucleotide is substituted with a non-fluorine group;
Alternatively, a nucleotide in which the hydroxy group at the 2' position of the ribose group of the nucleotide is substituted with a non-fluorine group is 2'-alkoxy modified nucleotide, 2'-substituted alkoxy modified nucleotide, 2'-alkyl modified nucleotide, 2'-substituted one selected from alkyl modified nucleotides, 2'-amino modified nucleotides, 2'-substituted amino modified nucleotides and 2'-deoxy nucleotides, and the nucleotide analog is selected from isonucleotides, LNA, ENA, cET, UNA and GNA is one to be
Alternatively, the siRNA complex of claim 12 , wherein each non-fluoro-modified nucleotide is a methoxy-modified nucleotide, and the methoxy-modified nucleotide refers to a nucleotide in which the 2'-hydroxy group of the ribose group is replaced with methoxy. body.

From the 5' end to the 3' end, nucleotides at positions 5, 7, 8 and 9 of nucleotide sequence I in the sense strand of the siRNA are fluoro-modified nucleotides, and nucleotides at the remaining positions of the sense strand of the siRNA are methoxy-modified nucleotides, and nucleotides at positions 2, 6, 8, 9, 14 and 16 of nucleotide sequence II in the antisense strand of the siRNA from the 5' end to the 3' end are fluoro-modified nucleotides; the nucleotides at the remaining positions of the antisense strand of the siRNA are methoxy-modified nucleotides;
Alternatively, from the 5' end to the 3' end, nucleotides at positions 5, 7, 8 and 9 of nucleotide sequence I in the sense strand of the siRNA are fluoro-modified nucleotides, and the remaining positions of the sense strand of the siRNA are The nucleotides are methoxy-modified nucleotides, the nucleotides at positions 2, 6, 14 and 16 of nucleotide sequence II in the antisense strand of the siRNA are fluoro-modified nucleotides from the 5' end to the 3' end, and the siRNA the nucleotides at the remaining positions of the antisense strand are methoxy-modified nucleotides,
Alternatively, from the 5' end to the 3' end, the nucleotides at positions 7, 8 and 9 of nucleotide sequence I in the sense strand of the siRNA are fluoro-modified nucleotides, and the nucleotides at the remaining positions of the sense strand of the siRNA are It is a methoxy-modified nucleotide, and nucleotides at positions 2, 6, 14 and 16 of nucleotide sequence II in the antisense strand of the siRNA are fluoro-modified nucleotides from the 5' end to the 3' end, and the siRNA antisense 2. The siRNA complex of claim 1 , wherein the nucleotides at the remaining positions of the strand are methoxy-modified nucleotides.

In the siRNA, at least one phosphate ester group is a thiophosphate ester group, and the thiophosphate ester group is
between the first and second nucleotides from the 5' end of the sense strand,
between the second and third nucleotides from the 5' end of the sense strand,
between the first and second nucleotides from the 3′ end of the sense strand,
between the second and third nucleotides from the 3′ end of the sense strand,
Between the first and second nucleotides from the 5' end of the antisense strand,
between the second and third nucleotides from the 5' end of the antisense strand,
selected from the group consisting of between the 1st and 2nd nucleotides from the 3' end of the antisense strand, and between the 2nd and 3rd nucleotides from the 3' end of the antisense strand 2. The siRNA complex of claim 1 , present in association with at least one.

the 5'-terminal nucleotide of the antisense strand of the siRNA is a 5'-phosphate nucleotide or a 5'-phosphate analog modified nucleotide;
Alternatively, the 5'-phosphate nucleotide is a nucleotide having a structure represented by formula (2), and the 5'-phosphate analog modified nucleotide is any one of formulas (3) to (6) selected from nucleotides having the structure shown in

2. The siRNA complex of claim 1 , wherein R is selected from H, OH, a methoxy group or fluorine, and Base represents a base and is selected from A, U, C, G or T.

The siRNA is siPCSKa1-M1, siPCSKa1-M2, siPCSKa1-M3, siPCSKa2-M1, siPCSKa2-M2, siPCSKa2-M3, siPCSKb1-M1, siPCSKb1-M2, siPCSKb1-M3, siPCSKb2-M1, siPCSKb 2-M2, siPCSKb2- M3, siPCSKc1-M1, siPCSKc1-M2, siPCSKc1-M3, siPCSKc2-M1, siPCSKc2-M2, siPCSKc2-M3, siPCSKd1-M1, siPCSKd1-M2, siPCSKd1-M3, siPCSKd2-M1, siPCSKd2- M2, siPCSKd2-M3, siPCSKe1-M1 siPCSKe1-M2 siPCSKe1-M3 siPCSKe2-M1 siPCSKe2-M2 siPCSKe2-M3 siPCSKf1-M1 siPCSKf1-M2 Any of siPCSKf2-M3 is one, or
The siRNA is siPCSKa1-M1S, siPCSKa1-M2S, siPCSKa1-M3S, siPCSKa2-M1S, siPCSKa2-M2S, siPCSKa2-M3S, siPCSKb1-M1S, siPCSKb1-M2S, siPCSKb1-M3S, siPCSKb2- M1S, siPCSKb2-M2S, siPCSKb2- M3S, siPCSKc1-M1S, siPCSKc1-M2S, siPCSKc1-M3S, siPCSKc2-M1S, siPCSKc2-M2S, siPCSKc2-M3S, siPCSKd1-M1S, siPCSKd1-M2S, siPCSKd1-M3S, siPCSKd2-M 1S, siPCSKd2-M2S, siPCSKd2-M3S, siPCSKe1-M1S siPCSKe1-M2S siPCSKe1-M3S siPCSKe2-M1S Either iPCSKf2-M2S or siPCSKf2-M3S is one or said siRNA is 1-M3SP1, siPCSKa2-M1SP1, siPCSKa2-M2SP1, siPCSKa2-M3SP1, siPCSKb1-M1P1, siPCSKb1-M2P1, siPCSKb1-M3P1, siPCSKb2-M1P1, siPCSKb2-M2P1, siPCSKb2-M3P1, siPCSKb1-M1SP1, siPCS Kb1-M2SP1, siPCSKb1-M3SP1, siPCSKb2-M1SP1, siPCSKb2- M2SP1, siPCSKb2-M3SP1, siPCSKc1-M1P1, siPCSKc1-M2P1, siPCSKc1-M3P1, siPCSKc2-M1P1, siPCSKc2-M2P1, siPCSKc2-M3P1, siPCSKc1-M1SP1, siPCSKc1-M2SP1 , siPCSKc1-M3SP1, siPCSKc2-M1SP1, siPCSKc2-M2SP1, siPCSKc2-M3SP1, siPCSKd1-M1P1, siPCSKd1-M2P1, siPCSKd1-M3P1, siPCSKd2-M1P1, siPCSKd2-M2P1, siPCSKd2-M3P1, siPCSKd1-M1SP1, siPCSKd1-M2SP1, siPCS Kd1-M3SP1, siPCSKd2-M1SP1, siPCSKd2-M2SP1, siPCSKd2- M3SP1, siPCSKe1-M1P1, siPCSKe1-M2P1, siPCSKe1-M3P1, siPCSKe2-M1P1, siPCSKe2-M2P1, siPCSKe2-M3P1, siPCSKe1-M1SP1, siPCSKe1-M2SP1, siPCSKe1-M3SP1 , siPCSKe2-M1SP1, siPCSKe2-M2SP1, siPCSKe2-M3SP1, siPCSKf1-M1P1, siPCSKf1-M2P1, siPCSKf1-M3P1, siPCSKf2-M1P1, siPCSKf2-M2P1, siPCSKf2-M3P1, siPCSKf1-M1SP1, siPCSKf1-M2SP1, siPCSKf1-M3SP1, siPCS Either Kf2-M1SP1, siPCSKf2-M2SP1 or siPCSKf2-M3SP1 The siRNA complex of claim 1 , which is one.

A composition for treating and/or preventing a disease or physiological condition caused by abnormal expression of the PCSK9 gene , comprising the siRNA complex according to any one of claims 1 to 17 .

A composition for suppressing PCSK9 gene expression in hepatocytes , comprising the siRNA complex according to any one of claims 1 to 17 .