Double-stranded oligonucleotide and conjugate for inhibiting expression of c3 genes, and uses thereof

Abstract

The present application relates to the field of drugs, and in particular to a double-stranded oligonucleotide and conjugate for inhibiting the expression of C3 genes, and uses thereof. The double-stranded oligonucleotide or the conjugate comprising same can obviously inhibit the expression of C3 mRNA in HepG2 cells, can obviously inhibit the expression of C3 mRNA in animal bodies, can obviously reduce the level of C3 protein in the serum of cynomolgus monkeys, the inhibition effect being able to last for 42 days, can obviously inhibit alternative complement pathways of the serum of cynomolgus monkeys, but has no effect on classical complement pathways of the serum, can reduce the levels of C3 mRNA and protein of CFA-IgA mice, can reduce C3 deposition in a renal tissue, and can ameliorate the disease course of a patient with C3 deposition in the renal tissue. Additionally, no obvious abnormality is found in hematology and blood biochemistry of animals. The double-stranded oligonucleotide or the conjugate comprising same is beneficial to alleviating, preventing and / or treating diseases or disorders mediated by C3 genes.

Description

Double-stranded oligonucleotides and conjugates for inhibiting C3 gene expression and their uses Technical Field

[0001] The present disclosure relates to the field of medicine, and in particular to double-stranded oligonucleotides and conjugates for inhibiting C3 gene expression and their uses. Background Art

[0002] The complement system is an important component of the body's innate immunity. It is composed of more than 50 soluble proteins and membrane-bound proteins. It plays an important role in the body's life activities such as resisting foreign substances, cell lysis, inflammatory response, dissolution of immune complexes, clearing apoptotic cells, and enhancing humoral immune response.

[0003] Complement activation occurs via three major pathways: the classical pathway, the alternative pathway, and the mannose-binding lectin (MBL) pathway.

[0004] Because the complement system plays a crucial role in defending the body against foreign substances, clearing immune complexes, and participating in adaptive immunity, complement system abnormalities are closely associated with a variety of diseases, including kidney disease and autoimmune disorders. For example, atypical hemolytic uremic syndrome (aHUS) is characterized by microangiopathy, hemolytic anemia, thrombocytopenia, and acute renal failure. Over 50% of aHUS cases are associated with mutations or polymorphisms in complement regulatory factors such as Factor H, Factor I, MCP, C4BP, Factor B, and C3, as well as the production of Factor H autoantibodies. These mutations or polymorphisms often occur in heterozygous forms and affect the secretion and function of these proteins. Furthermore, membranoproliferative type II glomerulonephritis (MPGN) is a severe kidney disease characterized by electron-dense deposits and proliferation of glomerular basement membrane and mesangial cells, as seen under an electron microscope. People who are deficient in Factor H or C3 are susceptible to this disease. Furthermore, the body's production of autoantibodies against the C3 convertase in the alternative pathway (also called nephritic factor C3NeF) is also associated with MPGN. Glomerulonephritis is a disease in which immune complexes in the renal blood vessels cause excessive complement activation, resulting in kidney damage. C3 and its regulatory proteins, Factor I and Factor H, are closely associated with the development and progression of glomerulonephritis. The absence of Factor I and Factor H can lead to disrupted C3 regulation. Furthermore, C1q knockout mice can produce autoantibodies, are unable to effectively clear apoptotic cells, and are susceptible to immune complex-induced glomerulonephritis.

[0005] Targets currently under development within the classical, lectin, and alternative complement pathways include C1 (C1q, C1r, C1s), MASPs, C2, C3, C5, and C6. Targeting C3, a key complement protein, several companies are developing peptide drugs (such as AMY-101 and APL-1) or recombinant enzymes (CB2782) for the treatment of diseases such as paronychia (PNH), chronic obstructive pulmonary disease (COPD), and AMD. However, few drugs targeting C3, a key complement protein, are currently under development for the treatment of kidney diseases.

[0006] Therefore, providing a double-stranded oligonucleotide to inhibit the expression of C3 gene is of great significance for improving the course of patients with renal tissue C3 deposition, and alleviating, preventing and / or treating diseases or conditions mediated by C3 gene.

[0007] Summary of the Invention

[0008] In view of this, the present disclosure provides double-stranded oligonucleotides, conjugates, and uses thereof for inhibiting C3 gene expression.

[0009] In order to achieve the above-mentioned invention objectives, the present disclosure provides the following technical solutions:

[0010] In a first aspect of the present disclosure, the present disclosure provides a double-stranded oligonucleotide for inhibiting C3 gene expression, wherein the double-stranded oligonucleotide comprises a sense strand and an antisense strand;

[0011] In the 5' to 3' direction, nucleotides 2 to 19 of the antisense strand comprise a region complementary to a C3 RNA transcript, wherein the complementary region comprises at least 17 consecutive nucleotides of any one of SEQ ID Nos. 213 to 424, or a nucleotide sequence that differs from the consecutive nucleotides by 1, 2, or 3 nucleotides;

[0012] The sense strand comprises at least 17 nucleotides, and the sense strand and the antisense strand are complementary or substantially complementary to form a duplex region; the substantial complementarity means that the mismatch between the sense strand and the antisense strand in the duplex region is no more than 3 nucleotides.

[0013] In some optional embodiments of the present disclosure, the antisense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 213 to 424 and has 0, 1, 2 or 3 nucleotide differences.

[0014] In some optional embodiments of the present disclosure, the antisense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 213 to 424 having 0 or 1 nucleotide difference.

[0015] In some specific embodiments of the present disclosure, the antisense strand is selected from or comprises any one of the nucleotide sequences of SEQ ID No. 213 to 424.

[0016] In some optional embodiments of the present disclosure, the sense strand comprises at least 17 consecutive nucleotides in any one of the nucleotide sequences of SEQ ID NOs. 1 to 212, or a nucleotide sequence having 1, 2, or 3 nucleotide differences from the consecutive nucleotides.

[0017] In some optional embodiments of the present disclosure, the sense strand comprises a nucleotide sequence of any one of SEQ ID NOs. 1 to 212 and has 0, 1, 2 or 3 nucleotide differences.

[0018] In some optional embodiments of the present disclosure, the sense strand comprises a nucleotide sequence of any one of SEQ ID NOs. 1 to 212 and has 0 or 1 nucleotide difference.

[0019] In some specific embodiments of the present disclosure, the sense strand is selected from or comprises any one of the nucleotide sequences of SEQ ID NOs. 1 to 212.

[0020] In some specific embodiments of the present disclosure, the antisense strand is selected from or comprises any one of nucleotide sequences of SEQ ID Nos. 213 to 424, and the sense strand is selected from or comprises any one of nucleotide sequences of SEQ ID Nos. 1 to 212.

[0021] In some optional embodiments of the present disclosure, the antisense strand comprises a nucleotide sequence of any one of SEQ ID NO. 241, 242, 285, 357 or 403 and having 0 or 1 nucleotide difference;

[0022] In some specific embodiments of the present disclosure, the antisense strand comprises any one of the nucleotide sequences of SEQ ID NOs. 241, 242, 285, 357 and 403.

[0023] In some optional embodiments of the present disclosure, the sense strand comprises a nucleotide sequence of any one of SEQ ID NOs. 29, 30, 73, 145, and 191 having 0 or 1 nucleotide difference;

[0024] In some optional embodiments of the present disclosure, the sense strand comprises any one of the nucleotide sequences of SEQ ID NOs. 29, 30, 73, 145 and 191.

[0025] In some specific embodiments of the present disclosure, the antisense strand comprises any one of SEQ ID NOs. 241, 242, 285, 357 and 403, and the sense strand comprises any one of SEQ ID NOs. 29, 30, 73, 145 and 191.

[0026] In some specific embodiments of the present disclosure, the double-stranded oligonucleotide comprises one or more of the following:

[0027] An antisense strand having the nucleotide sequence shown in SEQ ID NO. 241 or having 0 or 1 nucleotide difference therefrom, and a sense strand having the nucleotide sequence shown in SEQ ID NO. 29 or having 0 or 1 nucleotide difference therefrom;

[0028] An antisense strand having the nucleotide sequence of SEQ ID NO. 242 or having 0 or 1 nucleotide difference therefrom, and a sense strand having the nucleotide sequence of SEQ ID NO. 30 or having 0 or 1 nucleotide difference therefrom;

[0029] An antisense strand having the nucleotide sequence of SEQ ID NO. 285 or having 0 or 1 nucleotide difference therefrom, and a sense strand having the nucleotide sequence of SEQ ID NO. 73 or having 0 or 1 nucleotide difference therefrom;

[0030] An antisense strand having a nucleotide sequence as shown in SEQ ID NO. 357 or having 0 or 1 nucleotide difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID NO. 145 or having 0 or 1 nucleotide difference therefrom;

[0031] The antisense strand has the nucleotide sequence shown in SEQ ID NO. 403 or has 0 or 1 nucleotide difference therefrom, and the sense strand has the nucleotide sequence shown in SEQ ID NO. 191 or has 0 or 1 nucleotide difference therefrom.

[0032] In some specific embodiments of the present disclosure, the double-stranded oligonucleotide comprises one or more of the following:

[0033] The antisense strand has the nucleotide sequence shown in SEQ ID NO. 241, and the sense strand has the nucleotide sequence shown in SEQ ID NO. 29;

[0034] The antisense strand has the nucleotide sequence shown in SEQ ID NO. 242, and the sense strand has the nucleotide sequence shown in SEQ ID NO. 30;

[0035] The antisense strand has the nucleotide sequence shown in SEQ ID NO. 285, and the sense strand has the nucleotide sequence shown in SEQ ID NO. 73;

[0036] The antisense strand has the nucleotide sequence shown in SEQ ID NO.357, and the sense strand has the nucleotide sequence shown in SEQ ID NO.145;

[0037] The antisense strand has the nucleotide sequence shown in SEQ ID NO.403, and the sense strand has the nucleotide sequence shown in SEQ ID NO.191;

[0038] In some specific embodiments of the present disclosure, the double-stranded oligonucleotide comprises:

[0039] The antisense strand has the nucleotide sequence shown in SEQ ID NO. 241 or has 0 or 1 nucleotide difference therefrom, and the sense strand has the nucleotide sequence shown in SEQ ID NO. 29 or has 0 or 1 nucleotide difference therefrom.

[0040] In some specific embodiments of the present disclosure, the double-stranded oligonucleotide comprises:

[0041] The antisense strand has the nucleotide sequence shown in SEQ ID NO.241, and the sense strand has the nucleotide sequence shown in SEQ ID NO.29.

[0042] In some specific embodiments of the present disclosure, each nucleotide in the double-stranded oligonucleotide is independently selected from modified or unmodified nucleotides.

[0043] In some specific embodiments of the present disclosure, substantially all nucleotides in the double-stranded oligonucleotide are independently selected from modified nucleotides. Wherein, "substantially all nucleotides are selected from modified nucleotides" means that most but not all of the nucleotides in the double-stranded oligonucleotide are modified nucleotides, and may contain no more than 5, 4, 3, 2 or 1 unmodified nucleotides.

[0044] In some specific embodiments of the present disclosure, all nucleotides in the double-stranded oligonucleotide are independently selected from modified nucleotides.

[0045] In some specific embodiments of the present disclosure, each nucleotide in the double-stranded oligonucleotide is independently selected from the following modified nucleotides:

[0046] 2'-halogenated modified nucleotides, 2'-deoxy modified nucleotides, 2'-O- optionally substituted C 1-6 Alkyl-modified nucleotides, 2'-O-(CH2) n-O-R1 modified nucleotides, 2'-amino modified nucleotides, abasic nucleotides or quasi-nucleotides; the quasi-nucleotides are selected from one or more of peptide nucleic acid (PNA), morpholino (MNA), bridged nucleic acid (BNA), locked nucleic acid (LNA), glycol nucleic acid / glycerol nucleic acid (GNA), threose nucleic acid (TNA) or unlocked nucleic acid (UNA);

[0047] wherein n is selected from 1 or 2, R1 is selected from optionally substituted C 1-6 Alkyl or optionally substituted C 1-6 Alkoxy; if R1 contains a substituent, the substituent is selected from halogen, C 1-6 alkoxy, hydroxy or amino.

[0048] In some specific embodiments of the present disclosure, each nucleotide in the sense strand and / or antisense strand is independently selected from modified or unmodified nucleotides.

[0049] In some optional embodiments of the present disclosure, all nucleotides in the sense strand and / or antisense strand are substantially selected from modified nucleotides. Wherein, "substantially all nucleotides are selected from modified nucleotides" means that most but not all of the nucleotides in the strand are modified nucleotides, and may contain no more than 5, 4, 3, 2 or 1 unmodified nucleotides.

[0050] In some specific embodiments of the present disclosure, all nucleotides in the sense strand and / or the antisense strand are selected from modified nucleotides.

[0051] In some specific embodiments of the present disclosure, each nucleotide in the sense strand and / or the antisense strand is independently selected from the following modified nucleotides:

[0052] 2'-halogenated modified nucleotides, 2'-deoxy modified nucleotides, 2'-O-optionally substituted C1-6 alkyl modified nucleotides, 2'-O-(CH2) n-O-R1 modified nucleotides, 2'-amino modified nucleotides, abasic nucleotides, or quasi-nucleotides; the quasi-nucleotides are selected from one or more of peptide nucleic acid (PNA), morpholino (MNA), bridged nucleic acid (BNA), locked nucleic acid (LNA), glycol nucleic acid / glycerol nucleic acid (GNA), threose nucleic acid (TNA), or unlocked nucleic acid (UNA);

[0053] wherein said n is selected from 1 or 2;

[0054] The R1 is selected from optionally substituted C 1-6 Alkyl or optionally substituted C 1-6 Alkoxy; if R1 contains a substituent, the substituent is selected from halogen, C 1-6 alkoxy, hydroxy or amino.

[0055] Among them, the structural formula of nucleotide is: Base represents a nucleoside base, and the nucleoside base on each nucleotide is independently selected from uracil U, thymine T, cytosine C, adenine A or guanine G.

[0056] In the present disclosure, a 2'-halogenated modified nucleotide refers to a nucleotide in which the hydroxyl group at the 2' position of the ribose group is substituted by a halogen atom, such as a 2'-fluoro modified nucleotide.

[0057] In some specific embodiments of the present disclosure, the 2'-halogenated modified nucleotides are selected from 2'-fluoro-modified nucleotides.

[0058] In the present disclosure, a 2'-deoxy modified nucleotide refers to a nucleotide in which the hydroxyl group at the 2' position of the ribose group is replaced by a hydrogen atom.

[0059] In the present disclosure, 2'-O-(CH2) n -R1 modified nucleotides refer to nucleotides in which the hydrogen atom on the 2' hydroxyl group of the ribose group is replaced by -(CH2) n -R1 substitution. Wherein, when n is selected from 1, the 2'-O-(CH2) n -R1 modified nucleotides are selected from 2'-O-ethoxymethyl modified nucleotides or 2'-O-2,2,2-trifluoroethoxymethyl modified nucleotides. When n is selected from 2, the 2'-O-(CH2)n-R1 modified nucleotides are selected from 2'-O-methoxyethyl modified nucleotides (also known as 2'-O-moe modified nucleotides).

[0060] In some alternative embodiments of the present disclosure, 2'-O-(CH2) n -R1 modified nucleotides are selected from 2'-O-methoxyethyl modified nucleotides, 2'-O-ethoxymethyl modified nucleotides or 2'-O-2,2,2-trifluoroethoxymethyl modified nucleotides.

[0061] In some alternative embodiments of the present disclosure, 2'-O-(CH2) n -R1 modified nucleotides are selected from 2'-O-methoxyethyl modified nucleotides.

[0062] As used herein, "optionally substituted" is used to define a variable, which may be unsubstituted or substituted.

[0063] In some optional embodiments of the present disclosure, each nucleotide in the sense strand and / or the antisense strand is independently selected from the following modified nucleotides:

[0064] 2'-O-methyl modified nucleotides, 2'-fluoro modified nucleotides, 2'-O-methoxyethyl modified nucleotides or 2'-deoxy modified nucleotides.

[0065] In some specific embodiments of the present disclosure, each nucleotide in the sense strand and / or the antisense strand is independently selected from the following modified nucleotides:

[0066] 2'-O-methyl modified nucleotides, 2'-fluoro modified nucleotides or 2'-O-methoxyethyl modified nucleotides.

[0067] In some specific embodiments of the present disclosure, each nucleotide in the sense strand and / or antisense strand is independently selected from the following modified nucleotides:

[0068] 2'-O-methyl modified nucleotides, 2'-fluoro modified nucleotides or 2'-deoxy modified nucleotides.

[0069] In some specific embodiments of the present disclosure, the double-stranded oligonucleotide contains at least one 2'-O-methoxyethyl modified nucleotide.

[0070] In some specific embodiments of the present disclosure, the length of the sense strand is selected from 18 to 21 nucleotides, and the length of the antisense strand is selected from 19 to 23 nucleotides.

[0071] In some specific embodiments of the present disclosure, the sense strand is 17-21 nucleotides long, and the antisense strand is 19-23 nucleotides long.

[0072] In some specific embodiments of the present disclosure, the sense strand length is selected from 19 nucleotides, and the antisense strand length is selected from 21 nucleotides.

[0073] In some specific embodiments of the present disclosure, the antisense strand contains at least one 2'-O-methoxyethyl-modified nucleotide; and the sense strand contains at most one 2'-O-methoxyethyl-modified nucleotide.

[0074] In some specific embodiments of the present disclosure, the sense strand does not contain 2'-O-methoxyethyl modified nucleotides, and the antisense strand contains one 2'-O-methoxyethyl modified nucleotide. Each nucleotide in the sense strand is independently selected from a 2'-fluoro modified nucleotide or a 2'-O-methyl modified nucleotide; each nucleotide in the antisense strand is independently selected from a 2'-fluoro modified nucleotide, a 2'-O-methyl modified nucleotide, or a 2'-O-methoxyethyl modified nucleotide, and the antisense strand contains one 2'-O-methoxyethyl modified nucleotide.

[0075] In some specific embodiments of the present disclosure, the sense strand contains a 2'-O-methoxyethyl modified nucleotide, and the antisense strand contains a 2'-O-methoxyethyl modified nucleotide. Each nucleotide in the sense strand is independently selected from a 2'-fluoro modified nucleotide, a 2'-O-methyl modified nucleotide, or a 2'-O-methoxyethyl modified nucleotide; and each nucleotide in the antisense strand is independently selected from a 2'-fluoro modified nucleotide, a 2'-O-methyl modified nucleotide, or a 2'-O-methoxyethyl modified nucleotide.

[0076] In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, at least three nucleotides in positions 7 to 10 of the nucleotide sequence in the positive strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides or 2'-O-methoxyethyl-modified nucleotides.

[0077] In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, at least three nucleotides at positions 7 to 10 of the nucleotide sequence in the positive strand are selected from 2'-fluoro-modified nucleotides, at most two nucleotides at positions 5, 12, 13, and 18 are selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides.

[0078] In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, at least three nucleotides at positions 7 to 10 of the nucleotide sequence in the positive strand are selected from 2'-fluoro-modified nucleotides, at most one nucleotide at positions 5, 12, 13, and 18 are selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides.

[0079] In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, the 7th to 10th nucleotides of the nucleotide sequence in the positive strand are selected from 2'-fluoro-modified nucleotides, at most one nucleotide at positions 5, 12, 13, and 18 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides.

[0080] In some specific embodiments of the present disclosure, the modification of the sense strand is selected from one of the following (SS-1) to (SS-5):

[0081] (SS-1) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 in the nucleotide sequence of the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides;

[0082] (SS-2) in the sense strand, nucleotides 7 to 10 are selected from 2'-fluoro-modified nucleotides, nucleotide 5 is selected from 2'-O-methoxyethyl-modified nucleotides, and nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides, in the direction from the 5' end to the 3' end;

[0083] (SS-3) in the sense strand, nucleotides 7 to 10 are selected from 2'-fluoro-modified nucleotides, nucleotide 12 is selected from 2'-O-methoxyethyl-modified nucleotides, and nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides, in the direction from the 5' end to the 3' end;

[0084] (SS-4) in the sense strand, nucleotides 7 to 10 are selected from 2'-fluoro-modified nucleotides, nucleotide 13 is selected from 2'-O-methoxyethyl-modified nucleotides, and the remaining nucleotides are independently selected from 2'-O-methyl-modified nucleotides, in the direction from the 5' end to the 3' end.

[0085] (SS-5) In the direction from the 5' end to the 3' end, the 7th to 10th nucleotides of the nucleotide sequence in the positive strand are selected from 2'-fluoro-modified nucleotides, the 18th nucleotide is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides.

[0086] In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, at least four nucleotides in positions 2, 6, 9 to 12, 14 and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides or 2'-O-methoxyethyl-modified nucleotides.

[0087] In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, at least four nucleotides of positions 2, 6, 9 to 12, 14 and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at positions 2, 6, 14 and 16 are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides or 2'-O-methoxyethyl-modified nucleotides.

[0088] In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, at least four nucleotides of positions 2, 6, 14 and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, and any one nucleotide from positions 9 to 12 is selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides or 2'-O-methoxyethyl-modified nucleotides.

[0089] In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, at least four nucleotides of positions 2, 6, 14 and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, and any one nucleotide at positions 9 to 12 is selected from 2'-fluoro-modified nucleotides, at least one nucleotide at positions 8 and 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides.

[0090] In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, at least four nucleotides of positions 2, 6, 14 and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, and any one of the nucleotides at positions 9 to 12 is selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 or position 15 is selected from a 2'-O-methoxyethyl-modified nucleotide, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides.

[0091] In some specific embodiments of the present disclosure, the modification of the antisense strand is selected from one of the following (AS-1) to (AS-8):

[0092] (AS-1) in the nucleotide sequence of the antisense strand, in the direction from the 5' end to the 3' end, the nucleotides at positions 2, 6, 9, 14, and 16 are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides;

[0093] (AS-2) in the nucleotide sequence of the antisense strand, in the direction from the 5' end to the 3' end, the nucleotides at positions 2, 6, 10, 14, and 16 are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides;

[0094] (AS-3) in the nucleotide sequence of the antisense strand, in the direction from the 5' end to the 3' end, the nucleotides at positions 2, 6, 11, 14, and 16 are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides;

[0095] (AS-4) in the nucleotide sequence of the antisense strand, in the direction from the 5' end to the 3' end, the nucleotides at positions 2, 6, 12, 14, and 16 are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides;

[0096] (AS-5) in the nucleotide sequence of the antisense strand, in the direction from the 5' end to the 3' end, the nucleotides at positions 2, 6, 9, 14, and 16 are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides;

[0097] (AS-6) in the nucleotide sequence of the antisense strand, in the direction from the 5' end to the 3' end, the nucleotides at positions 2, 6, 10, 14, and 16 are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides;

[0098] (AS-7) In the direction from the 5' end to the 3' end, the 2nd, 6th, 11th, 14th and 16th nucleotides of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the 15th nucleotide is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides.

[0099] (AS-8) In the direction from the 5' end to the 3' end, the 2nd, 6th, 12th, 14th and 16th nucleotides of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the 15th nucleotide is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides.

[0100] In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, at least three nucleotides in positions 7 to 10 of the nucleotide sequence in the sense strand are 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides or 2'-O-methoxyethyl-modified nucleotides;

[0101] At least four nucleotides in positions 2, 6, 9 to 12, 14 and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro modified nucleotides, and the nucleotides in the remaining positions are independently selected from 2'-O-methyl modified nucleotides or 2'-O-methoxyethyl modified nucleotides.

[0102] In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, at least three nucleotides in positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are independently selected from 2'-O-methyl-modified nucleotides or 2'-O-methoxyethyl-modified nucleotides;

[0103] At least four nucleotides in positions 2, 6, 9 to 12, 14 and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides in positions 2, 6, 14 and 16 are selected from 2'-fluoro-modified nucleotides, and the nucleotides in the remaining positions are independently selected from 2'-O-methyl-modified nucleotides or 2'-O-methoxyethyl-modified nucleotides.

[0104] In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, at least three nucleotides in positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, at most two nucleotides in positions 5, 12, 13, and 18 are selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides in the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0105] At least four nucleotides at positions 2, 6, 9 to 12, 14 and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at positions 2, 6, 14 and 16 are selected from 2'-fluoro-modified nucleotides, at least one nucleotide at positions 8 and 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides.

[0106] In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, at most one nucleotide at positions 5, 12, 13, and 18 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0107] At least four of the nucleotides at positions 2, 6, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, and any one of the nucleotides at positions 9 to 12 is selected from 2'-fluoro-modified nucleotides, at least one of the nucleotides at positions 8 and 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides.

[0108] In some specific embodiments of the present disclosure, the modifications of the sense strand and the antisense strand in the double-stranded oligonucleotide are selected from one of the following (ds-1) to (ds-8):

[0109] (ds-1) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 in the nucleotide sequence of the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0110] The nucleotides at positions 2, 6, 9, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0111] (ds-2) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 in the nucleotide sequence of the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0112] The nucleotides at positions 2, 6, 10, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0113] (ds-3) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 in the nucleotide sequence of the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0114] The nucleotides at positions 2, 6, 11, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0115] (ds-4) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0116] The nucleotides at positions 2, 6, 12, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0117] (ds-5) nucleotides 7 to 10 in the nucleotide sequence of the sense strand, from the 5' end to the 3' end, are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0118] The nucleotides at positions 2, 6, 9, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0119] (ds-6) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 in the nucleotide sequence of the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0120] The nucleotides at positions 2, 6, 10, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0121] (ds-7) nucleotides 7 to 10 in the nucleotide sequence of the sense strand, from the 5' end to the 3' end, are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0122] The nucleotides at positions 2, 6, 11, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0123] (ds-8) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0124] The nucleotides at positions 2, 6, 12, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; (ds-9) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 5 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0125] The nucleotides at positions 2, 6, 9, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0126] (ds-10) in the sense strand, from the 5' end to the 3' end, nucleotides 7 to 10 are selected from 2'-fluoro-modified nucleotides, the 5th nucleotide is selected from a 2'-O-methoxyethyl-modified nucleotide, and the remaining nucleotides are selected from 2'-O-methyl-modified nucleotides;

[0127] The nucleotides at positions 2, 6, 10, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0128] (ds-11) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 5 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0129] The nucleotides at positions 2, 6, 11, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0130] (ds-12) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 5 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0131] The nucleotides at positions 2, 6, 9, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0132] (ds-13) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 5 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0133] The nucleotides at positions 2, 6, 10, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0134] (ds-14) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 5 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0135] The nucleotides at positions 2, 6, 11, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0136] (ds-15) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 12 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0137] The nucleotides at positions 2, 6, 9, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0138] (ds-16) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 12 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0139] The nucleotides at positions 2, 6, 10, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0140] (ds-17) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 12 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0141] The nucleotides at positions 2, 6, 11, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0142] (ds-18) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 12 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0143] The nucleotides at positions 2, 6, 9, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0144] (ds-19) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 12 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0145] The nucleotides at positions 2, 6, 10, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0146] (ds-20) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 12 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0147] The nucleotides at positions 2, 6, 11, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0148] (ds-21) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 13 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0149] The nucleotides at positions 2, 6, 9, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0150] (ds-22) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 13 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0151] The nucleotides at positions 2, 6, 10, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0152] (ds-23) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 13 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0153] The nucleotides at positions 2, 6, 11, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0154] (ds-24) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 13 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0155] The nucleotides at positions 2, 6, 9, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0156] (ds-25) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 13 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0157] The nucleotides at positions 2, 6, 10, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0158] (ds-26) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 13 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0159] The nucleotides at positions 2, 6, 11, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0160] (ds-27) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 18 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0161] The nucleotides at positions 2, 6, 9, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0162] (ds-28) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 18 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0163] The nucleotides at positions 2, 6, 10, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0164] (ds-29) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 18 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0165] The nucleotides at positions 2, 6, 11, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0166] (ds-30) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 18 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0167] The nucleotides at positions 2, 6, 9, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0168] (ds-31) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 18 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0169] The nucleotides at positions 2, 6, 10, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0170] (ds-32) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 18 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0171] The nucleotides at positions 2, 6, 11, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides. In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0172] At least four of the nucleotides at positions 2, 6, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, and any one of the nucleotides at positions 9 to 12 is selected from 2'-fluoro-modified nucleotides, at least one of the nucleotides at positions 8 and 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides.

[0173] In some specific embodiments of the present disclosure, in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0174] At least four of the nucleotides at positions 2, 6, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, and any one of the nucleotides at positions 9 to 12 is selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 or position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides.

[0175] In some specific embodiments of the present disclosure, the modifications of the sense strand and the antisense strand in the double-stranded oligonucleotide are selected from one of the following (ds-1) to (ds-8):

[0176] (ds-1) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 in the nucleotide sequence of the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0177] The nucleotides at positions 2, 6, 9, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0178] (ds-2) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 in the nucleotide sequence of the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0179] The nucleotides at positions 2, 6, 10, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0180] (ds-3) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 in the nucleotide sequence of the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0181] The nucleotides at positions 2, 6, 11, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0182] (ds-4) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0183] The nucleotides at positions 2, 6, 12, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0184] (ds-5) nucleotides 7 to 10 in the nucleotide sequence of the sense strand, from the 5' end to the 3' end, are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0185] The nucleotides at positions 2, 6, 9, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0186] (ds-6) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 in the nucleotide sequence of the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0187] The nucleotides at positions 2, 6, 10, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0188] (ds-7) nucleotides 7 to 10 in the nucleotide sequence of the sense strand, from the 5' end to the 3' end, are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0189] The nucleotides at positions 2, 6, 11, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0190] (ds-8) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides;

[0191] The nucleotides at positions 2, 6, 12, 14 and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl modified nucleotides.

[0192] In some specific embodiments of the present disclosure, the sense strand and / or the antisense strand independently comprises one or more phosphorothioate internucleotide linkages.

[0193] In some specific embodiments of the present disclosure, the sense strand comprises two consecutive phosphorothioate internucleotide linkages at the 5' terminus.

[0194] In some specific embodiments of the present disclosure, the antisense strand comprises two consecutive phosphorothioate internucleotide linkages at the 5' end and two consecutive phosphorothioate internucleotide linkages at the 3' end.

[0195] In some specific embodiments of the present disclosure, the sense strand comprises two phosphorothioate internucleotide linkages at the 5' terminus; the antisense strand comprises two phosphorothioate internucleotide linkages at the 5' terminus and two phosphorothioate internucleotide linkages at the 3' terminus.

[0196] In some optional embodiments of the present disclosure, the sense strand or the antisense strand comprises a 3' overhang of at least 1 nucleotide.

[0197] In some alternative embodiments of the present disclosure, the antisense strand comprises a 3' overhang of at least 1 nucleotide.

[0198] In some alternative embodiments of the present disclosure, the antisense strand comprises a 3' overhang of 2 nucleotides.

[0199] In some specific embodiments of the present disclosure, each nucleotide in the double-stranded oligonucleotide is independently selected from modified nucleotides;

[0200] Wherein, the sense strand is selected from any one of nucleotide sequences in SEQ ID No. 887 to 907; and / or the antisense strand is selected from any one of nucleotide sequences in SEQ ID No. 908 to 943;

[0201] The positive chain: from the 5' end to the 3' end,

[0202] SEQ ID No.887: CmsGmsAmAmGmCmUfCfAfUfGmAmAmUmAmUmAmUmUm

[0203] SEQ ID No.888: CmsGmsAmAmG(moe)CmUfCfAfUfGmAmAmUmAmUmAmUmUm

[0204] SEQ ID No.889: CmsGmsAmAmGmCmUfCfAfUfGmA(moe)AmUmAmUmAmUmUm

[0205] SEQ ID No.890: CmsGmsAmAmGmCmUfCfAfUfGmAmA(moe)UmAmUmAmUmUm

[0206] SEQ ID No.891:CmsGmsAmAmGmCmUfCfAfUfGmAmAmUmAmUmAmT(moe)Um

[0207] SEQ ID No.892:CmsAmsGmAmGmAmAfAfUfUfCmUmAmCmUmAmCmAmUm

[0208] SEQ ID No.893: CmsAmsGmAmG(moe)AmAfAfUfUfCmUmAmCmUmAmCmAmUm

[0209] SEQ ID No.894:CmsAmsGmAmGmAmAfAfUfUfCmT(moe)AmCmUmAmCmAmUm

[0210] SEQ ID No.895:CmsAmsGmAmGmAmAfAfUfUfCmUmA(moe)CmUmAmCmAmUm

[0211] SEQ ID No.896:CmsAmsGmAmGmAmAfAfUfUfCmUmAmCmUmAmCmA(moe)Um

[0212] SEQ ID No.897:GmsAmsGmAmAmUmUfGfCfUfUmCmAmUmAmCmAmAmAm

[0213] SEQ ID No.898:GmsAmsGmAmA(moe)UmUfGfCfUfUmCmAmUmAmCmAmAmAm

[0214] SEQ ID No.899:GmsAmsGmAmAmUmUfGfCfUfUmC(moe)AmUmAmCmAmAmAm

[0215] SEQ ID No.900:GmsAmsGmAmAmUmUfGfCfUfUmCmA(moe)UmAmCmAmAmAm

[0216] SEQ ID No.901:GmsAmsGmAmAmUmUfGfCfUfUmCmAmUmAmCmAmA(moe)Am

[0217] SEQ ID No.902:CmsAmsAmCmUmCmAfCfCfUfGmUmAmAmUmAmAmAmUm

[0218] SEQ ID No.903: CmsAmsAmCmT(moe)CmAfCfCfUfGmUmAmAmUmAmAmAmUm

[0219] SEQ ID No.904: CmsAmsAmCmUmCmAfCfCfUfGmT(moe)AmAmUmAmAmAmUm

[0220] SEQ ID No.905: CmsAmsAmCmUmCmAfCfCfUfGmUmA(moe)AmUmAmAmAmUm

[0221] SEQ ID No.906: CmsAmsAmCmUmCmAfCfCfUfGmUmAmAmUmAmAmA(moe)Um

[0222] SEQ ID No.907: AmsAmsAmUmUmCmUfAfCfUfAmCmAmUmCmUmAmUmAm

[0223] The antisense strand: in the direction from the 5' end to the 3' end,

[0224] SEQ ID No.908: AmsAfsUmAmUmAfUmUmCfAmUmGmAmGfC(moe)UfUmCmGmsUmsAm

[0225] SEQ ID No.909: AmsAfsUmAmUmAfUmUmCmAfUmGmAmGfC(moe)UfUmCmGmsUmsAm

[0226] SEQ ID No.910: AmsAfsUmAmUmAfUmUmCmAmUfGmAmGfC(moe)UfUmCmGmsUmsAm

[0227] SEQ ID No.911: AmsAfsUmAmUmAfUmT(moe)CmAfUmGmAmGfCmUfUmCmGmsUmsAm

[0228] SEQ ID No.912: AmsUfsGmUmAmGfUmAmGfAmAmUmUmUfC(moe)UfCmUmGmsUmsAm

[0229] SEQ ID No.913: AmsUfsGmUmAmGfUmAmGmAfAmUmUmUfC(moe)UfCmUmGmsUmsAm

[0230] <h2 style=";text-align:left;direction:ltr">SEQ ID No.914:AmsUfsGmUmAmGfUmAmGmAmAfUmUmUfC(moe)UfCmUmGmsUmsAm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0231] <h2 style=";text-align:left;direction:ltr"> SEQ ID No. 915: AmsUfsGmUmAmGfUmA(moe)GmAfAmUmUmUfCmUfCmUmGmsUmsAm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0232] <h2 style=";text-align:left;direction:ltr"> SEQ ID No.916:UmsUfsUmGmUmAfUmGmAfAmGmCmAmAfT(moe)UfCmUmCmsCmsUm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0233] <h2 style=";text-align:left;direction:ltr"> SEQ ID No.917:UmsUfsUmGmUmAfUmGmAmAfGmCmAmAfT(moe)UfCmUmCmsCmsUm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0234] <h2 style=";text-align:left;direction:ltr"> SEQ ID No.918:UmsUfsUmGmUmAfUmGmAmAmGfCmAmAfT(moe)UfCmUmCmsCmsUm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0235] <h2 style=";text-align:left;direction:ltr"> SEQ ID No.919:UmsUfsUmGmUmAfUmG(moe)AmAfGmCmAmAfUmUfCmUmCmsCmsUm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0236] <h2 style=";text-align:left;direction:ltr"> SEQ ID No. 920: AmsUfsUmUmAmUfUmAmCfAmGmGmUmGfA(moe)GfUmUmGmsAmsUm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0237] <h2 style=";text-align:left;direction:ltr"> SEQ ID No. 921: AmsUfsUmUmAmUfUmAmCmAfGmGmUmGfA(moe)GfUmUmGmsAmsUm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0238] <h2 style=";text-align:left;direction:ltr"> SEQ ID No. 922:AmsUfsUmUmAmUfUmAmCmAmGfGmUmGfA(moe)GfUmUmGmsAmsUm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0239] <h2 style=";text-align:left;direction:ltr"> SEQ ID No. 923: AmsUfsUmUmAmUfUmA(moe)CmAfGmGmUmGfAmGfUmUmGmsAmsUm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0240] <h2 style=";text-align:left;direction:ltr"> SEQ ID No.924:AmsAfsUmAmUmAfUmT(moe)CmAmUfGmAmGfCmUfUmCmGmsUmsAm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0241] <h2 style=";text-align:left;direction:ltr">SEQ ID No.925:AmsUfsGmUmAmGfUmA(moe)GmAmAfUmUmUfCmUfCmUmGmsUmsAm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0242] <h2 style=";text-align:left;direction:ltr"> SEQ ID No. 926: AmsUfsUmUmAmUfUmA(moe)CmAmGfGmUmGfAmGfUmUmGmsAmsUm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0243] <h2 style=";text-align:left;direction:ltr"> SEQ ID No.927:AmsUfsGmUmAmGfUmAmGmAmAmUfUmUfC(moe)UfCmUmGmsUmsAm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0244] <h2 style=";text-align:left;direction:ltr"> SEQ ID No.928:UmsAfsUmAmGmAfUmGmUmAmGmUfAmGfAmAfUmUmUmsCmsUm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0245] <h2 style=";text-align:left;direction:ltr"> SEQ ID No. 929: UmsAfsUmAmGmAfUmGmUfAmGmUmAmGfA(moe)AfUmUmUmsCmsUm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0246] <h2 style=";text-align:left;direction:ltr"> SEQ ID No. 930: UmsAfsUmAmGmAfUmGmUmAmGmUfAmGfA(moe)AfUmUmUmsCmsUm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0247] <h2 style=";text-align:left;direction:ltr"> SEQ ID No.931:AmsAfsUmAmUmAfUmUmCfAmUmGmAmGfCmUfUmCmGmsUmsAm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0248] <h2 style=";text-align:left;direction:ltr"> SEQ ID No.932:AmsAfsUmAmUmAfUmUmCmAmUmGfAmGfCmUfUmCmGmsUmsAm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0249] <h2 style=";text-align:left;direction:ltr"> SEQ ID No.933:AmsAfsUmAmUmAfUmUmCmAmUmGfAmGfC(moe)UfUmCmGmsUmsAm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0250] <h2 style=";text-align:left;direction:ltr"> SEQ ID No.934:AmsUfsUmUmAmUfUmAmCfAmGmGmUmGfAmGfUmUmGmsAmsUm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0251] <h2 style=";text-align:left;direction:ltr"> SEQ ID No.935:AmsUfsUmUmAmUfUmAmCmAmGmGfUmGfAmGfUmUmGmsAmsUm<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0252] SEQ ID No.936: AmsUfsUmUmAmUfUmAmCmAmGmGfUmGfA(moe)GfUmUmGmsAmsUm

[0253] SEQ ID No.937: AmsUfsGmUmAmGfUmAmGfAmAmUmUmUfCmUfCmUmGmsUmsAm

[0254] SEQ ID No.938: AmsUfsGmUmAmGfUmAmGmAmAmUfUmUfCmUfCmUmGmsUmsAm

[0255] SEQ ID No.939: UmsAfsUmAmGmAfUmGmUfAmGmUmAmGfAmAfUmUmUmsCmsUm

[0256] SEQ ID No.940: UmsUfsUmGmUmAfUmGmAfAmGmCmAmAfUmUfCmUmCmsCmsUm

[0257] SEQ ID No.941: UmsUfsUmGmUmAfUmGmAmAmGmCfAmAfUmUfCmUmCmsCmsUm

[0258] SEQ ID No.942: UmsUfsUmGmUmAfUmGmAfAmGmCmAmAfU(moe)UfCmUmCmsCmsUm

[0259] SEQ ID No.943: UmsUfsUmGmUmAfUmGmAmAmGmCfAmAfU(moe)UfCmUmCmsCmsUm

[0260] Among them, C represents cytidine-3'-phosphate, G represents guanosine-3'-phosphate, U represents uridine-3'-phosphate, A represents adenosine-3'-phosphate, and T represents thymidine-3'-phosphate; m represents that the nucleotide adjacent to the left of the letter m is a 2'-O-methyl-modified nucleotide; f represents that the nucleotide adjacent to the left of the letter f is a 2'-fluoro-modified nucleotide; (moe) represents that the nucleotide adjacent to the left of the combination mark (moe) is a 2'-O-methoxyethyl-modified nucleotide; s represents that the two nucleotides are connected by a phosphorothioate diester bond.

[0261] Among them, the structural formula of U is The structural formula of T(moe) is In this article, T(moe) is regarded as a modification of U.

[0262] It is particularly pointed out that in the context of the present disclosure, since sequences in which all nucleotides are modified cannot pass the inspection normally and result in the inability to generate a sequence listing, all sequences involving modifications in the present disclosure cannot be presented in the sequence listing, and all sequences are subject to the description in the specification.

[0263] In some specific embodiments of the present disclosure, the double-stranded oligonucleotide comprises one or more of the following:

[0264] In some specific embodiments of the present disclosure, the sense strand is selected from any one of SEQ ID No. 887, 892, 897, 902 and 907; and / or

[0265] The antisense strand is selected from any one of the nucleotide sequences of SEQ ID No. 908, 912, 914, 920, 927 to 943.

[0266] In some specific embodiments of the present disclosure, the double-stranded oligonucleotide comprises one or more of the following:

[0267] In some specific embodiments of the present disclosure, the double-stranded oligonucleotide comprises the following group:

[0268] In some specific embodiments of the present disclosure, the double-stranded oligonucleotide is selected from siRNA.

[0269] In a second aspect of the present disclosure, the present disclosure provides a double-stranded oligonucleotide for inhibiting C3 gene expression, wherein the double-stranded oligonucleotide comprises a sense strand and an antisense strand, and each nucleotide in the double-stranded oligonucleotide is independently selected from modified nucleotides;

[0270] wherein positions 2 to 19 of the antisense strand comprise at least 17 consecutive nucleotides of any one of SEQ ID Nos. 908 to 943, or a nucleotide sequence having 1, 2, or 3 nucleotide base differences from the consecutive nucleotides;

[0271] The sense strand comprises at least 17 nucleotides, and the sense strand and the antisense strand are complementary or substantially complementary to form a duplex region; the substantial complementarity means that the mismatch between the sense strand and the antisense strand in the duplex region is no more than 3 nucleotides.

[0272] In some optional embodiments of the present disclosure, the antisense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 908 to 943 and has 0, 1, 2 or 3 nucleotide base differences.

[0273] In some optional embodiments of the present disclosure, the antisense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 908 to 943 having 0 or 1 nucleotide base difference.

[0274] In some optional embodiments of the present disclosure, the antisense strand comprises any one of the nucleotide sequences of SEQ ID Nos. 908 to 943.

[0275] In some optional embodiments of the present disclosure, the sense strand comprises at least 17 consecutive nucleotides in any one of SEQ ID Nos. 887 to 907, or a nucleotide sequence having 1, 2 or 3 nucleotide base differences from the consecutive nucleotides.

[0276] In some optional embodiments of the present disclosure, the sense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 887 to 907 and has 0, 1, 2 or 3 nucleotide base differences.

[0277] In some optional embodiments of the present disclosure, the sense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 887 to 907 having 0 or 1 nucleotide base difference.

[0278] In some optional embodiments of the present disclosure, the sense strand comprises any one of the nucleotide sequences of SEQ ID Nos. 887 to 907.

[0279] In some optional embodiments of the present disclosure, the antisense strand comprises any one of the nucleotide sequences of SEQ ID Nos. 908 to 943, and the sense strand comprises any one of the nucleotide sequences of SEQ ID Nos. 887 to 907.

[0280] In some optional embodiments of the present disclosure, the double-stranded oligonucleotide comprises one or more of the following groups:

[0281] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 908 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 887 and having 0 or 1 nucleotide base difference therefrom;

[0282] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 909 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 887 and having 0 or 1 nucleotide base difference therefrom;

[0283] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 910 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 887 and having 0 or 1 nucleotide base difference therefrom;

[0284] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 909 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 888 and having 0 or 1 nucleotide base difference therefrom;

[0285] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 909 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 889 and having 0 or 1 nucleotide base difference therefrom;

[0286] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 909 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 890 and having 0 or 1 nucleotide base difference therefrom;

[0287] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 909 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 891 and having 0 or 1 nucleotide base difference therefrom;

[0288] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 911 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 888 and having 0 or 1 nucleotide base difference therefrom;

[0289] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 911 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 889 and having 0 or 1 nucleotide base difference therefrom;

[0290] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 911 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 890 and having 0 or 1 nucleotide base difference therefrom;

[0291] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 911 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 891 and having 0 or 1 nucleotide base difference therefrom;

[0292] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 912 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 892 and having 0 or 1 nucleotide base difference therefrom;

[0293] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 913 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 892 and having 0 or 1 nucleotide base difference therefrom;

[0294] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 914 and having 0 or 1 nucleotide base difference therefrom; and a sense strand having a nucleotide sequence as shown in SEQ ID No. 892 or having 0 or 1 nucleotide difference therefrom;

[0295] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 913 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 893 and having 0 or 1 nucleotide base difference therefrom;

[0296] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 913 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 894 and having 0 or 1 nucleotide base difference therefrom;

[0297] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 913 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 895 and having 0 or 1 nucleotide base difference therefrom;

[0298] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 913 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 896 and having 0 or 1 nucleotide base difference therefrom;

[0299] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 915 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 893 and having 0 or 1 nucleotide base difference therefrom;

[0300] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 915 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 894 and having 0 or 1 nucleotide base difference therefrom;

[0301] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 915 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 895 and having 0 or 1 nucleotide base difference therefrom;

[0302] an antisense strand having a nucleotide sequence as shown in SEQ ID No. 915 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 896 and having 0 or 1 nucleotide base difference therefrom;

[0303] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 916 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 897 and having 0 or 1 nucleotide base difference therefrom;

[0304] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 917 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 897 and having 0 or 1 nucleotide base difference therefrom;

[0305] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 918 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 897 and having 0 or 1 nucleotide base difference therefrom;

[0306] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 917 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 898 and having 0 or 1 nucleotide base difference therefrom;

[0307] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 917 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 899 and having 0 or 1 nucleotide base difference therefrom;

[0308] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 917 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 900 and having 0 or 1 nucleotide base difference therefrom;

[0309] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 917 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 901 and having 0 or 1 nucleotide base difference therefrom;

[0310] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 919 and having 0 or 1 nucleotide base difference therefrom; and a sense strand having a nucleotide sequence as shown in SEQ ID No. 898 and having 0 or 1 nucleotide base difference therefrom;

[0311] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 919 and having 0 or 1 nucleotide base difference therefrom; and a sense strand having a nucleotide sequence as shown in SEQ ID No. 899 and having 0 or 1 nucleotide base difference therefrom;

[0312] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 919 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 900 and having 0 or 1 nucleotide base difference therefrom;

[0313] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 919 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 901 and having 0 or 1 nucleotide base difference therefrom;

[0314] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 920 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom;

[0315] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 921 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom;

[0316] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 922 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom;

[0317] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 921 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 903 and having 0 or 1 nucleotide base difference therefrom;

[0318] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 921 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 904 and having 0 or 1 nucleotide base difference therefrom;

[0319] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 921 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 905 and having 0 or 1 nucleotide base difference therefrom;

[0320] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 921 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 906 and having 0 or 1 nucleotide base difference therefrom;

[0321] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 923 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 903 and having 0 or 1 nucleotide base difference therefrom;

[0322] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 923 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 904 and having 0 or 1 nucleotide base difference therefrom;

[0323] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 923 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 905 and having 0 or 1 nucleotide base difference therefrom;

[0324] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 923 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 906 and having 0 or 1 nucleotide base difference therefrom;

[0325] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 910 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 888 and having 0 or 1 nucleotide base difference therefrom;

[0326] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 910 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 890 and having 0 or 1 nucleotide base difference therefrom;

[0327] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 924 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 890 and having 0 or 1 nucleotide base difference therefrom;

[0328] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 924 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 891 and having 0 or 1 nucleotide base difference therefrom;

[0329] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 914 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 895 and having 0 or 1 nucleotide base difference therefrom;

[0330] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 925 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 896 and having 0 or 1 nucleotide base difference therefrom;

[0331] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 922 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 905 and having 0 or 1 nucleotide base difference therefrom;

[0332] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 926 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 906 and having 0 or 1 nucleotide base difference therefrom;

[0333] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 927 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 892 and having 0 or 1 nucleotide base difference therefrom;

[0334] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 928 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom;

[0335] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 929 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom;

[0336] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 930 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom;

[0337] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 931 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 887 and having 0 or 1 nucleotide base difference therefrom;

[0338] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 932 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 887 and having 0 or 1 nucleotide base difference therefrom;

[0339] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 933 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 887 and having 0 or 1 nucleotide base difference therefrom;

[0340] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 934 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom;

[0341] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 935 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom;

[0342] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 936 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom;

[0343] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 937 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 892 and having 0 or 1 nucleotide base difference therefrom;

[0344] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 938 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 892 and having 0 or 1 nucleotide base difference therefrom;

[0345] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 939 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom;

[0346] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 940 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 897 and having 0 or 1 nucleotide base difference therefrom;

[0347] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 941 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 897 and having 0 or 1 nucleotide base difference therefrom;

[0348] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 942 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 897 and having 0 or 1 nucleotide base difference therefrom;

[0349] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 943 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 897 and having 0 or 1 nucleotide base difference therefrom.

[0350] In some specific embodiments of the present disclosure, the double-stranded oligonucleotide comprises one or more of the following:

[0351] In some optional embodiments of the present disclosure, the antisense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 908, 912, 914, 920, 927 to 943 and has 0, 1, 2 or 3 nucleotide base differences.

[0352] In some optional embodiments of the present disclosure, the antisense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 908, 912, 914, 920, 927 to 943 and has 0 or 1 nucleotide base difference.

[0353] In some optional embodiments of the present disclosure, the antisense strand comprises any one of the nucleotide sequences of SEQ ID Nos. 908, 912, 914, 920, 927-943.

[0354] In some optional embodiments of the present disclosure, the sense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 887, 892, 897, 902 and 907 and has 0, 1, 2 or 3 nucleotide base differences.

[0355] In some optional embodiments of the present disclosure, the sense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 887, 892, 897, 902 and 907 and has 0 or 1 nucleotide base difference.

[0356] In some optional embodiments of the present disclosure, the sense strand comprises any one of the nucleotide sequences of SEQ ID Nos. 887, 892, 897, 902 and 907.

[0357] In some specific embodiments of the present disclosure, the antisense strand comprises any one of SEQ ID Nos. 908, 912, 914, 920, 927 to 943, and the sense strand comprises any one of SEQ ID Nos. 887, 892, 897, 902 and 907.

[0358] In some optional embodiments of the present disclosure, the double-stranded oligonucleotide comprises one or more of the following groups:

[0359] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 908 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 887 and having 0 or 1 nucleotide base difference therefrom;

[0360] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 931 and having 0 or 1 nucleotide base difference therefrom; and a sense strand having a nucleotide sequence as shown in SEQ ID No. 887 and having 0 or 1 nucleotide base difference therefrom;

[0361] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 932 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 887 and having 0 or 1 nucleotide base difference therefrom;

[0362] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 933 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 887 and having 0 or 1 nucleotide base difference therefrom;

[0363] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 912 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 892 and having 0 or 1 nucleotide base difference therefrom;

[0364] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 914 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 892 and having 0 or 1 nucleotide base difference therefrom;

[0365] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 927 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 892 and having 0 or 1 nucleotide base difference therefrom

[0366] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 937 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 892 and having 0 or 1 nucleotide base difference therefrom

[0367] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 938 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 892 and having 0 or 1 nucleotide base difference therefrom

[0368] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 940 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 897 and having 0 or 1 nucleotide base difference therefrom

[0369] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 941 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 897 and having 0 or 1 nucleotide base difference therefrom

[0370] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 942 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 897 and having 0 or 1 nucleotide base difference therefrom;

[0371] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 943 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 897 and having 0 or 1 nucleotide base difference therefrom;

[0372] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 920 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom

[0373] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 934 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom

[0374] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 935 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom;

[0375] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 936 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom;

[0376] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 928 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom;

[0377] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 929 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom;

[0378] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 930 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom

[0379] An antisense strand having a nucleotide sequence as shown in SEQ ID No. 939 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom.

[0380] In some specific embodiments of the present disclosure, the double-stranded oligonucleotide comprises one or more of the following:

[0381] In some optional embodiments of the present disclosure, the antisense strand comprises a nucleotide sequence shown in SEQ ID No. 914 and having 0, 1, 2 or 3 nucleotide base differences.

[0382] In some optional embodiments of the present disclosure, the antisense strand comprises a nucleotide sequence shown in SEQ ID No. 914 and having 0 or 1 nucleotide base difference.

[0383] In some specific embodiments of the present disclosure, the antisense strand comprises the nucleotide sequence shown in SEQ ID No.914.

[0384] In some optional embodiments of the present disclosure, the sense strand comprises the nucleotide sequence shown in SEQ ID No. 892 and has 0, 1, 2 or 3 nucleotide base differences.

[0385] In some optional embodiments of the present disclosure, the sense strand comprises the nucleotide sequence shown in SEQ ID No. 892 and having 0 or 1 nucleotide base difference.

[0386] In some specific embodiments of the present disclosure, the sense strand comprises the nucleotide sequence shown in SEQ ID No. 892.

[0387] In some optional embodiments of the present disclosure, the double-stranded oligonucleotide comprises:

[0388] An antisense strand having the nucleotide sequence shown in SEQ ID No. 914 or having 0 or 1 nucleotide base difference therefrom, and a sense strand having the nucleotide sequence shown in SEQ ID No. 892 or having 0 or 1 nucleotide base difference therefrom.

[0389] In some specific embodiments of the present disclosure, the double-stranded oligonucleotide comprises:

[0390] The antisense strand has the nucleotide sequence shown in SEQ ID No.914, and the sense strand has the nucleotide sequence shown in SEQ ID No.892.

[0391] In some specific embodiments of the present disclosure, the double-stranded oligonucleotide is selected from siRNA.

[0392] In a third aspect of the present disclosure, the present disclosure provides a conjugate comprising the double-stranded oligonucleotide according to the first aspect and / or the second aspect, and one or more gene delivery vectors.

[0393] In some alternative embodiments of the present disclosure, the gene delivery vehicle is selected from the group consisting of lipids, nanoparticles, polymers, and ligands.

[0394] In some optional embodiments of the present disclosure, the lipid comprises cholesterol or a cholesterol derivative; the polymer comprises an amphiphilic polymer or a membrane active polymer; and the ligand is selected from ligands capable of binding to cell surface receptors.

[0395] In some specific embodiments of the present disclosure, the gene delivery vector is selected from ligands capable of binding to cell surface receptors.

[0396] In some specific embodiments of the present disclosure, the ligand is specifically targeted to liver tissue.

[0397] In some specific embodiments of the present disclosure, the ligand is selected from asialoglycoprotein receptor ligand (ASGPR ligand). The binding of the ASGPR ligand to ASGPR can promote the specific targeting of the conjugate to hepatocytes and the internalization of the conjugate into hepatocytes.

[0398] In some specific embodiments of the present disclosure, the ASGPR ligand comprises galactose or a derivative thereof.

[0399] In some optional embodiments of the present disclosure, the galactose or its derivatives include but are not limited to galactose, galactosamine, N-formylgalactosamine, N-acetylgalactosamine, N-propionylgalactosamine, N-n-butyrylgalactosamine or N-isobutyrylgalactosamine.

[0400] In some optional embodiments of the present disclosure, the galactose or its derivative is selected from N-acetylgalactosamine (GalNAc).

[0401] In some specific embodiments of the present disclosure, the ASGPR ligand comprises N-acetylgalactosamine.

[0402] In some optional embodiments of the present disclosure, the ligand is conjugated to the 3' end of the sense strand, the 5' end of the sense strand, the 3' end of the antisense strand and / or the 5' end of the antisense strand.

[0403] In some specific embodiments of the present disclosure, the number of the ligands is selected from one, and one ligand is conjugated to the 3' end of the sense strand.

[0404] In some specific embodiments of the present disclosure, the double-stranded oligonucleotide is selected from siRAN, and the conjugate is selected from siRAN conjugate.

[0405] In some optional embodiments of the present disclosure, the conjugate comprises a structure as shown in formula (101):

[0406] Wherein, Nu represents the double-stranded oligonucleotide;

[0407] Each of the R ligand independently selected from ASGPR ligands;

[0408] The m is selected from 1, 2, 3 or 4.

[0409] In some optional embodiments of the present disclosure, when m is selected from 1, one R ligand conjugated to the 3' end of the sense strand, or an R ligand conjugated to the 5' end of the sense strand, or an R ligand conjugated to the 3' end of the antisense strand, or an R ligand conjugated to the 5' end of the antisense strand;

[0410] When m is selected from 2, the two R ligand conjugated to the 3' end of the sense strand and the 5' end of the sense strand, respectively; or two R ligand conjugated to the 3' end of the sense strand and the 3' end of the antisense strand respectively; or two R ligand conjugated to the 3' end of the sense strand and the 5' end of the antisense strand, respectively; or two R ligand conjugated to the 5' end of the sense strand and the 3' end of the antisense strand, respectively; or two R ligand conjugated to the 5' end of the sense strand and the 5' end of the antisense strand respectively; or two R ligand conjugated to the 3' end of the antisense strand and the 5' end of the antisense strand, respectively;

[0411] When m is selected from 3, the three R ligand conjugated to the 3' end of the sense strand, the 5' end of the sense strand and the 3' end of the antisense strand respectively; or three R ligand conjugated to the 3' end of the sense strand, the 5' end of the sense strand and the 5' end of the antisense strand respectively; or three R ligand conjugated to the 3' end of the sense strand, the 3' end of the antisense strand and the 5' end of the antisense strand respectively; or three R ligand conjugated to the 5' end of the sense strand, the 3' end of the antisense strand, and the 5' end of the antisense strand, respectively;

[0412] When m is selected from 4, the four R ligand They are conjugated to the 3' end of the sense strand, the 5' end of the sense strand, the 3' end of the antisense strand and the 5' end of the antisense strand respectively.

[0413] In some specific embodiments of the present disclosure, m is selected from 1.

[0414] In some specific embodiments of the present disclosure, m is selected from 1, one Rligand Conjugated to the 3' end of the sense strand.

[0415] In some optional embodiments of the present disclosure, m is selected from 2.

[0416] In some optional embodiments of the present disclosure, m is selected from 2, two R ligand conjugated to the 3' and 5' ends of the sense strand, respectively.

[0417] In some optional embodiments of the present disclosure, each of the R ligand Independently selected from the structure represented by formula (201) or a pharmaceutically acceptable salt or stereoisomer thereof:

[0418] Wherein, * represents the conjugation site of the ligand on the sense strand or the antisense strand;

[0419] Said j is selected from 1, 2, 3 or 4;

[0420] Each of said Z is independently selected from hydroxyl or thiol;

[0421] Each of said p is independently selected from 1, 2 or 3;

[0422] Each of said q is independently selected from 1, 2 or 3;

[0423] Each R2 is independently selected from H, optionally substituted C 1-6 Alkyl or optionally substituted C 1-6 alkoxy;

[0424] Each of said L is independently selected from optionally substituted C 2-20 Alkylene or R La and R Lb Independently selected from optionally substituted C 1-10 Alkylene, k is selected from 1, 2, 3, 4 or 5;

[0425] Each Y is independently selected from O, S, and NH.

[0426] In some optional embodiments of the present disclosure, m is selected from 1, 2 or 3.

[0427] In some specific embodiments of the present disclosure, m is selected from 3.

[0428] In some specific embodiments of the present disclosure, the Z is selected from hydroxyl.

[0429] In some specific embodiments of the present disclosure, the p is selected from 1.

[0430] In some specific embodiments of the present disclosure, q is selected from 1.

[0431] In some specific embodiments of the present disclosure, each R2 is selected from H.

[0432] In some optional embodiments of the present disclosure, each of said L is independently selected from optionally substituted C 2-10 Alkylene or Among them, R La and R Lb Independently selected from optionally substituted C 1-10 Alkylene, k is selected from 1, 2 or 3.

[0433] In some specific embodiments of the present disclosure, k is selected from 1.

[0434] In some optional embodiments of the present disclosure, each L is independently selected from

[0435] In some optional embodiments of the present disclosure, each L is independently selected from

[0436] In some specific embodiments of the present disclosure, L is selected from

[0437] In some specific embodiments of the present disclosure, L is selected from

[0438] In some specific embodiments of the present disclosure, Y is selected from O.

[0439] In some optional embodiments of the present disclosure, each R ligand Independently selected from the structure represented by formula (202) or a pharmaceutically acceptable salt or stereoisomer thereof:

[0440] In some optional embodiments of the present disclosure, each R ligand Independently selected from the structure represented by formula (CR01008×3) or its pharmaceutically acceptable salt or stereoisomer:

[0441] In some optional embodiments of the present disclosure, each Rligand is independently selected from the structure shown in formula (CR01013×3) or a pharmaceutically acceptable salt or stereoisomer thereof:

[0442] In some specific embodiments of the present disclosure, each R ligand Independently selected from the structure shown in Formula L96 or a pharmaceutically acceptable salt thereof:

[0443] In some embodiments of the present disclosure, the conjugate includes one or more of the following:

[0444] Exemplary explanation: "SEQ ID No. 892_(CR01008×3)" indicates that the ligand represented by the formula (CR01008×3) is conjugated to the 3' end of the sense strand represented by SEQ ID No. 892.

[0445] In some embodiments of the present disclosure, the conjugate includes one or more of the following:

[0446] Exemplary explanation: "SEQ ID No. 892_(CR01008×3)" indicates that the ligand represented by the formula (CR01008×3) is conjugated to the 3' end of the sense strand represented by SEQ ID No. 892.

[0447] In some specific embodiments of the present disclosure, the conjugate includes the following group:

[0448] Here, "SEQ ID No.892_(CR01008×3)" indicates that the ligand represented by the formula (CR01008×3) is conjugated to the 3' end of the positive strand represented by SEQ ID No.892.

[0449] In the fourth aspect of the present disclosure, the present disclosure provides use of the double-stranded oligonucleotide described in the first aspect, and / or the double-stranded oligonucleotide described in the second aspect, and / or the conjugate described in the third aspect in the preparation of a medicament for alleviating, preventing and / or treating a disease or condition mediated by the C3 gene.

[0450] In some optional embodiments of the present disclosure, the diseases or conditions mediated by the C3 gene include but are not limited to IgA nephropathy, atypical hemolytic uremic syndrome (aHUS), including paroxysmal nocturnal hemoglobinuria (PNH), C3 glomerulopathy, wolf nephritis and / or membranous nephritis.

[0451] In a fifth aspect of the present disclosure, the present disclosure provides a pharmaceutical composition comprising the double-stranded oligonucleotide of the first aspect, and / or the double-stranded oligonucleotide of the second aspect, and / or the conjugate of the third aspect.

[0452] In some optional embodiments of the present disclosure, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients or adjuvants.

[0453] As used herein, "pharmaceutical composition" may refer to a composition for use in treating a disease or in vitro cell culture experiments. When used in treating a disease, the term "pharmaceutical composition" generally refers to a unit dosage form and can be prepared by any of the methods well known in the pharmaceutical art. All methods include the step of combining the active ingredient with an excipient that constitutes one or more adjunct ingredients. Typically, the composition is prepared by uniformly and thoroughly combining the active siRNA with a liquid excipient, a finely divided solid excipient, or both.

[0454] As used herein, the term "pharmaceutically acceptable" means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients of the formulation and / or the mammal to be treated therewith. Preferably, the "pharmaceutically acceptable" herein refers to a substance approved by a federal regulatory agency or a state government or listed in the U.S. Pharmacopoeia or other generally recognized pharmacopeia for use in animals, particularly humans.

[0455] As used herein, the term "pharmaceutically acceptable excipient" may include any solvent, solid excipient, diluent, or other liquid excipient, etc., suitable for the particular intended dosage form. Except to the extent that any conventional excipient is incompatible with the siRNA of the present disclosure, such as by producing any adverse biological effect or interacting in a deleterious manner with any other component of the pharmaceutically acceptable composition, their use is also contemplated by the present disclosure.

[0456] In addition to any conventional excipients, to the extent that they are incompatible with the siRNA of the present disclosure, such as by producing any adverse biological effects or interacting in a deleterious manner with any other component of the pharmaceutically acceptable composition, their use is also contemplated by the present disclosure.

[0457] The pharmaceutical compositions disclosed herein include formulations suitable for parenteral administration. The formulations can be conveniently presented in unit dosage form and can be prepared by any method known in the pharmaceutical art. The amount of active ingredient that can be combined with excipients to prepare a single dose is generally the amount of siRNA that produces a therapeutic effect.

[0458] In a sixth aspect of the present disclosure, the present disclosure provides a drug comprising the pharmaceutical composition according to the fifth aspect.

[0459] In a seventh aspect of the present disclosure, the present disclosure provides a drug combination comprising the drug according to the sixth aspect and any other active ingredients.

[0460] In the eighth aspect of the present disclosure, the present disclosure provides a kit comprising the double-stranded oligonucleotide described in the first aspect, and / or the double-stranded oligonucleotide described in the second aspect, and / or the conjugate described in the third aspect, and / or the pharmaceutical composition described in the fifth aspect, and / or the drug described in the sixth aspect, and / or the drug combination described in the seventh aspect.

[0461] In the ninth aspect of the present disclosure, the present disclosure provides a method for inhibiting C3 gene expression, comprising administering to a subject the double-stranded oligonucleotide of the first aspect, and / or the double-stranded oligonucleotide of the second aspect, and / or the conjugate of the third aspect, and / or the pharmaceutical composition of the fifth aspect, and / or the drug of the sixth aspect, and / or the drug combination of the seventh aspect.

[0462] In some specific embodiments of the present disclosure, a method for inhibiting C3 expression in cells in vitro comprises contacting the cells with the double-stranded oligonucleotide of the first aspect, and / or the double-stranded oligonucleotide of the second aspect, and / or the conjugate of the third aspect, and / or the pharmaceutical composition of the fifth aspect, and / or the drug of the sixth aspect, and / or the drug combination of the seventh aspect.

[0463] In some optional embodiments of the present disclosure, the expression of the C3 gene in the cell is inhibited for a period of time sufficient to allow the mRNA transcript of the C3 gene to be degraded.

[0464] Of more practical significance is that, in the tenth aspect of the present disclosure, the present disclosure provides a method for alleviating, treating and / or preventing C3-mediated diseases or conditions, which comprises administering to a subject the double-stranded oligonucleotide described in the first aspect, and / or the double-stranded oligonucleotide described in the second aspect, and / or the conjugate described in the third aspect, and / or the pharmaceutical composition described in the fifth aspect, and / or the drug described in the sixth aspect, and / or the drug combination described in the seventh aspect.

[0465] In some specific embodiments of the present disclosure, the subject is a human.

[0466] In some specific embodiments of the present disclosure, the drug is administered at a dose of 0.01 mg / kg to 10 mg / kg or 0.5 mg / kg to 50 mg / kg.

[0467] In some specific embodiments of the present disclosure, the administration of the drug includes subcutaneous administration or intravenous administration.

[0468] In some specific embodiments of the present disclosure, the C3-mediated disease or disorder includes a disease associated with the mRNA level of C3 gene expression.

[0469] In some specific embodiments of the present disclosure, the disease or condition comprises IgA nephropathy, atypical hemolytic uremic syndrome, paroxysmal nocturnal hemoglobinuria (PNH), C3 glomerulopathy, wolf nephritis, or membranous nephritis.

[0470] The double-stranded oligonucleotides or conjugates provided herein can significantly inhibit C3 gene mRNA expression in HepG2 cells; significantly inhibit C3 gene mRNA expression in animals; significantly reduce C3 gene protein levels in cynomolgus monkey serum, with the inhibitory effect persisting up to day 42; significantly inhibit the alternative complement pathway in cynomolgus monkey serum, but have no effect on the classical complement pathway; reduce C3 gene mRNA and protein levels in CFA-IgA mice; reduce C3 deposition in renal tissue and improve the course of disease in patients with renal C3 deposition; and no significant abnormalities were observed in animal hematology or blood biochemistry. The double-stranded oligonucleotides or conjugates provided herein are useful for alleviating, preventing, and / or treating diseases or conditions mediated by the C3 gene. BRIEF DESCRIPTION OF THE DRAWINGS

[0471] FIG1 shows the inhibitory activity of C3 mRNA in BALB / c-HDI mice after administration of the siRNA conjugate described in Example 2;

[0472] FIG2 shows typical histopathological images of each test group at 200 times magnification of HE staining in Example 3;

[0473] FIG3 shows the inhibitory activity of C3 mRNA in HepG2 cells after administration of the siRNA conjugate described in this example in Example 4;

[0474] FIG4 shows the inhibitory activity of C3 mRNA in the liver of cynomolgus monkeys after administration of the siRNA conjugate described in this example in Example 5;

[0475] FIG5 shows the change in C3 protein levels in cynomolgus monkey serum after administration of the siRNA conjugate described in Example 6;

[0476] FIG6 shows the changes in serum complement alternative pathway activity in cynomolgus monkeys after administration of the siRNA conjugates described in Example 7;

[0477] FIG7 shows the changes in serum classical complement pathway activity in cynomolgus monkeys after administration of the siRNA conjugates described in this example in Example 7;

[0478] FIG8 shows the inhibitory activity of target genes in the liver of CFA-hIgA mice after administration of the siRNA conjugate described in this example in Example 8;

[0479] FIG9 shows the changes in C3 protein levels in the serum of CFA-hIgA mice after administration of the siRNA conjugate described in this example in Example 8;

[0480] FIG10 shows the changes in C3 deposition in the kidneys of CFA-hIgA mice after administration of the siRNA conjugates described in Example 8. DETAILED DESCRIPTION

[0481] The present disclosure discloses double-stranded oligonucleotides, conjugates, and their uses for inhibiting C3 gene expression. Those skilled in the art can refer to the contents herein and appropriately modify the process parameters to achieve these results. It is particularly important to note that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this disclosure. The methods and applications of the present disclosure have been described through preferred embodiments. It is apparent that relevant personnel can modify or appropriately alter and combine the methods and applications described herein without departing from the content, spirit, and scope of this disclosure to implement and apply the disclosed technology.

[0482] Explanation of terms

[0483] In this document, the terms “include” or “comprising” are open expressions, that is, including the contents specified in the present disclosure, but not excluding other contents.

[0484] As used herein, the terms "optionally," "optional," or "optionally" generally mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0485] As used herein, the term "small interfering RNA (siRNA)" refers to a double-stranded RNA of 17 to 25 nucleotides in length, comprising a sense strand and an antisense strand. siRNA mediates the targeted cleavage of RNA transcripts in the RISC pathway by forming an RNA-induced silencing complex (RISC). Specifically, siRNA directs the specific degradation of mRNA sequences through the known RNA interference (RNAi) process, inhibiting the translation of mRNA into amino acids and protein. For example, siRNA can regulate (e.g., inhibit) the expression of C3 in cells.

[0486] As used herein, the term "antisense strand (or guide strand)" includes a region that is substantially complementary to a target sequence, such as C3 mRNA. The "follower strand" refers to an iRNA strand that is substantially complementary to the antisense strand. The term "substantially complementary" refers to complete complementarity or at least partial complementarity, for example, the antisense strand is completely complementary to the target sequence or at least partially complementary. In the case of partial complementarity, mismatches can exist within the interior or terminal regions of the molecule, wherein the most tolerated mismatches exist within the terminal regions, for example, within 5, 4, 3 or 2 nucleotides of the 5' end and / or 3' end of the iRNA.

[0487] It should be noted that "at least a portion of the antisense strand is substantially complementary to an mRNA" means that the antisense strand comprises a polynucleotide that is substantially complementary to a continuous portion of an mRNA of interest (e.g., an mRNA encoding C3). Alternatively, if a polynucleotide is substantially non-interruptedly complementary to a portion of an mRNA encoding C3, then the antisense strand is complementary to at least a portion of the C3 mRNA.

[0488] As used herein, the term "target sequence" refers to a contiguous portion of the nucleotide sequence of an mRNA molecule formed during transcription of a gene encoding C3, including mRNAs that are products of RNA processing of the primary transcript. C3 can be within a cell, for example, a cell within a subject.

[0489] As used herein, the term "substantially complementary" means that the sense and antisense strands have no more than 3 nucleotides, no more than 2 nucleotides, or no more than 1 nucleotide mismatch in the duplex region, such as 3 nucleotides, 2 nucleotides, 1 nucleotide, or 0 nucleotides mismatch.

[0490] In this document, the terms "nucleotide difference" and "nucleotide base difference" are used interchangeably. A "nucleotide difference" between one nucleotide sequence and another nucleotide sequence refers to a change in the base type of the nucleotide at the same position in the former compared to the latter. For example, when a nucleotide base in the latter is A, and the corresponding nucleotide base at the same position in the former is U, C, G, or T, it is considered that there is a nucleotide difference at that position between the two nucleotide sequences. In some embodiments, when a nucleotide at the original position is replaced by an abasic nucleotide or its equivalent, it can also be considered that a nucleotide difference occurs at that position.

[0491] As used herein, the term "overhang" refers to at least one unpaired nucleotide protruding from the double-stranded oligonucleotide duplex structure, and also refers to the nucleotide sequence in the siRNA structure outside the double-stranded region. For example, a nucleotide overhang exists when the 3' end of one of the sense and / or antisense strands extends beyond the 5' end of the other strand, or when the 5' end of one of the sense and / or antisense strands extends beyond the 3' end of the other strand. The overhang may comprise at least one nucleotide, at least two nucleotides, at least three nucleotides, at least four nucleotides, at least five nucleotides, or more. The nucleotide overhang may comprise or be composed of nucleotide / nucleoside analogs, including deoxynucleotides / nucleosides. The overhang may be located on the sense strand, the antisense strand, or any combination thereof. Furthermore, the nucleotides of the overhang may appear at the 5' end, the 3' end, or both ends of the antisense or sense strand.

[0492] As used herein, the term "DEPC H2O" refers to ultrapure water (grade 1 water) that has been treated with diethylpyrocarbonate (DEPC) and sterilized by high temperature and high pressure.

[0493] As used herein, the term "subject" refers to any animal being examined, studied, or treated, and is not intended to limit the present disclosure to any particular type of subject. In some embodiments of the present disclosure, humans are preferred subjects, while in other embodiments, non-human animals are preferred subjects, including but not limited to mice, monkeys, ferrets, cows, sheep, goats, pigs, chickens, turkeys, dogs, cats, horses, and reptiles.

[0494] As used herein, the term "inhibiting the expression of the C3 gene" includes any level of inhibition of the C3 gene, for example, at least partial inhibition of C3 gene expression, such as inhibition of at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%. C3 gene expression can be assessed based on the level of any variable associated with C3 gene expression, for example, C3 mRNA or protein levels. Inhibition can be assessed by a decrease in the absolute or relative level of one or more of these variables compared to a control level. The control level can be any type of control level utilized in the art, e.g., a baseline level prior to administration, or a level determined from a similar subject, cell, or sample that has not been treated or has been treated with a control (e.g., a buffer-only control or a no active agent control).

[0495] In the present disclosure, unless otherwise specified, the siRNA sequences used in the present disclosure were commissioned to Kunshan Aotai Biotechnology Co., Ltd. for synthesis; the PCR primers used in the present disclosure were commissioned to Sangon Biotech (Shanghai) Co., Ltd. for synthesis; the liver cancer HepG2 cells used in the present disclosure were purchased from Wuhan Pronocell Life Science Co., Ltd.; the experimental animals BALB / c mice used in the present disclosure were purchased from Zhejiang Weitong Lihua Experimental Animal Technology Co., Ltd.; the CFA-hIgA nephropathy model mice used in the present disclosure were constructed by Peking University First Hospital; and the healthy crab-eating macaques used in the present disclosure were purchased from Guangzhou Landao Biotechnology Co., Ltd.

[0496] In the present disclosure, unless otherwise specified, the base composition and modification meanings of the present disclosure are as follows: capital letters A, U, G, C, and T respectively represent the base composition of the nucleotide, lowercase letter m indicates that the nucleotide adjacent to the left of the letter m is a 2'-O-methyl modified (also known as: 2'-methoxy modified) nucleotide; lowercase letter f indicates that the nucleotide adjacent to the left of the letter f is a 2'-fluoro modified nucleotide; lowercase letter d indicates that the nucleotide adjacent to the left of the letter d is a 2'-deoxy modified ribonucleic acid (also known as: deoxyribonucleic acid); (moe) indicates that the nucleotide adjacent to the left of the combination mark (moe) is a 2'-O-methoxyethyl modified nucleotide; lowercase letter s indicates that the two nucleotides adjacent to the left and right of the letter s are connected by a phosphorothioate bond.

[0497] In this disclosure, unless otherwise stated, the real-time PCR detection data of the in vivo activity experiments involved in this disclosure are all calculated using the ΔΔCt method to perform relative quantitative calculation of the target gene mRNA in each test group. The calculation method is summarized as follows:

[0498] ΔCt(test group) = Ct(test group target gene) – Ct(test group reference gene)

[0499] ΔCt(control group) = Ct(control group target gene) – Ct(control group internal reference gene)

[0500] ΔΔCt(test group)=ΔCt(test group)-ΔCt(control group average)

[0501] ΔΔCt(control group) = ΔCt(control group) - ΔCt(control group average)

[0502] The mRNA expression level of the target gene in the test group was normalized with the control group as the benchmark, and the remaining expression level of the target gene mRNA in the control group was defined as 100%.

[0503] Relative residual expression level of target gene mRNA in the test group = 2 -ΔΔCt (Test group) × 100%

[0504] Test group target gene mRNA inhibition rate = 100% - test group target gene mRNA relative expression level

[0505] The calculation method for the detection data of C3 protein at different time points in the in vivo activity experiment involved in this disclosure is summarized as follows:

[0506] Wherein, pre-dose represents the serum C3 protein concentration before administration, and Dx represents the serum C3 protein concentration on the xth day after administration.

[0507] The calculation method of the detection data of CCP and CAP activities at different time points in the in vivo activity experiments involved in this disclosure is summarized as follows:

[0508] Wherein, pre-dose represents the CCP activity or CAP activity in the serum before administration, and Dx represents the CCP activity or CAP activity in the serum on the xth day after administration.

[0509] In this disclosure, unless otherwise stated, the in vivo activity experimental data used in this disclosure are based on (X±STDEV) The experimental data were plotted and analyzed using GraphPadprism 8.0 software.

[0510] In the present disclosure, unless otherwise stated, the ratios of reagents used in the present disclosure are calculated on a volume basis (v / v).

[0511] Unless otherwise specified, all reagents used in the preparation of the compounds disclosed herein were purchased from Beijing Coupling Technology Co., Ltd., wherein the information of the main reagents is shown in Table 1.

[0512] Table 1 Main reagents

[0513] Among them, CPG stands for controlled pore glass (Controlled Pore Glass) carrier.

[0514] Unless otherwise stated, the reagents, consumables, and instruments used in the biological detection experiments of this disclosure were all commercially available products. The main reagents and consumables and their sources are shown in Table 2, and the main instruments and their sources are shown in Table 3.

[0515] Table 2 Main reagents and consumables

[0516] Table 3 Main instruments and equipment

[0517] The double-stranded oligonucleotides and conjugates for inhibiting C3 gene expression provided by the present disclosure, as well as the raw materials and reagents used in their uses, are all commercially available.

[0518] The present disclosure is further described below with reference to the following embodiments:

[0519] Preparation of compounds

[0520] Preparation Example 1 Preparation of Compound CR01008 and Compound CR01008Z

[0521] (1.1) Synthesis of compound CR01008

[0522] In this preparation example, the synthetic route of compound CR01008 is as follows:

[0523] (1.1.1) Synthesis of Compound 2

[0524] Compound 1 (trans-4-(Boc-amino)cyclohexylcarboxaldehyde, 10.0 g, 1.0 eq) and formaldehyde solution (8.9 g, 37 mass %, 2.4 eq) were dissolved in 33 ml of methanol, and 13 ml of a 45.3 mass % KOH aqueous solution was added dropwise. After the addition was complete, the mixture was stirred at 25°C for 30 minutes, then heated to 60°C and refluxed at 60°C for 2 hours. After the reaction was completed, the reaction solution was cooled to room temperature and evaporated to dryness under reduced pressure to obtain a crude white solid. A small amount of water was added to the crude product to slurry it, and the mixture was filtered to obtain compound 2 (9 g, yield 78.9%) as a white solid. MS-ESI (m / z) = 260 [M+H] + .

[0525] (1.1.2) Synthesis of compound 3

[0526] Compound 2 (9 g, 1 eq) prepared in step (1.1.1) was dissolved in 70 mL of 1,4-dioxane. A 4 M solution of hydrogen chloride in 1,4-dioxane (45 mL) was added, and the mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction solution was evaporated to dryness under reduced pressure to obtain compound 3 (6.8 g, 100% yield) as a white solid.

[0527] (1.1.3) Synthesis of compound 5

[0528] Compound 3 (1.8 g, 2.0 eq), compound 4 (5-[[(2R,3R,4R,5R,6R)-3-acetylamino-4,5-diacetoxy-6-(acetoxymethyl)-2-tetrahydropyranyl]oxy]pentanoic acid, 2.1 g, 1.0 eq), and DIEA (N,N-diisopropylethylamine, 3.5 g, 6.0 eq) prepared in step (1.1.2) were dissolved in 15 ml of DMF. HBTU (1.9 g, 1.1 eq) was added, and the mixture was stirred at 25°C under a N2 atmosphere for 3 hours. After completion of the reaction, the reaction solution was evaporated to dryness under reduced pressure and reverse-phase purified (22% acetonitrile in water) to obtain compound 5 (1.78 g, 64.4% yield) as a white solid. MS-ESI (m / z) = 589 [M+H] + .

[0529] (1.1.4) Synthesis of Compound 6

[0530] Compound 5 (1.54 g, 1.0 eq) prepared in step (1.1.3) was dissolved in 15 ml of pyridine. The reaction system was cooled to 0°C using an ice-water bath and DMTrCl (4,4'-dimethoxytriphenylmethane chloride, 1.32 g, 1.5 eq) was added at 0°C. The reaction was allowed to react at 25°C for 3 hours. 15 ml of methanol was added to the reaction solution to quench the reaction. After completion of the reaction, the reaction solution was evaporated to dryness under reduced pressure and reverse-phase purified (60% acetonitrile in water) to obtain compound 6 (1 g, 42.7% yield) as a yellow solid. MS-ESI (m / z) = 891 [M+H] + .

[0531] (1.1.5) Synthesis of compound CR01008

[0532] Compound 6 (1.08 g, 1.0 eq) prepared in step (1.1.4) was dissolved in 20 ml of anhydrous dichloromethane. DCI (115 mg, 0.8 eq) and compound 7 (bis(diisopropylamino)(2-cyanoethoxy)phosphine, 732 mg, 2.1 eq) were added, respectively. The atmosphere was purged with nitrogen three times, and the mixture was stirred at 25°C for 2 hours. After completion of the reaction, 20 ml of saturated aqueous sodium bicarbonate solution was added to the reaction solution, and the mixture was extracted three times with 20 ml of dichloromethane (3×20 ml). The organic phases were combined and evaporated to dryness under reduced pressure. After reverse purification (72 vol% acetonitrile in water), the mixture was dried under vacuum for 12 hours to obtain compound CR01008 (1 g, 76.0% yield) as a white powder. MS-ESI (m / z) = 1091 [M+Na] + .

[0533] 1H NMR(400MHz, DMSO-d6)δ1.05(d,J=6.7Hz,6H).1.14(d,J=6.7Hz,6H),1.37–1.17(m,5H),1.60–1.40(m,6H),1.68–1.62(m,1H),1.80(s,3H), 1.80(s,3H),1.92(s,3H),2.02(s,5H),2.13(s,3H),2.71(t,J=5.9Hz,2H),2.79(d,J=8.4Hz,1H),2.87(d,J=8.4Hz,1H),3.36(s,1H),3.58– 3.39(m,3H),3.69–3.60(m,2H),3.75(s,7H),3.90(dt,J=11.2,8.8Hz,1H),4.05(s,3H),4.51(d,J=8.4Hz,1H),4.99(dd,J=11.3,3.4Hz,1H) ,5.24(d,J=3.4Hz,1H),5.78(s,1H),6.93–6.87(m,4H),7.35–7.21(m,7H),7.44–7.37(m,2H),7.66(d,J=7.8Hz,1H),7.84(d,J=9.2Hz,1H).

[0534] (1.2) Synthesis of compound CR01008Z

[0535] In this preparation example, the synthetic route of compound CR01008Z is as follows:

[0536] (1.2.1) Synthesis of compound 9

[0537] Compound 6 (500 mg) prepared in step (1.1.4) was dissolved in 10 ml of dichloromethane, and compound 8 (succinic anhydride, 112 mg), DMAP (6.8 mg), and TEA (226.2 mg) were added. The atmosphere was purged with nitrogen three times, and the mixture was stirred at 25°C for 16 hours. Flash purification was performed to obtain compound 9 (300 mg, yield 53.6%). MS-ESI (m / z) = 1013 [M+Na] + .

[0538] (1.2.2) Synthesis of compound CR01008Z

[0539] Compound 9 (50 mg), aminoCPG (1.25 g, 80 μmol / g, 0.1 mmol), HBTU (27 mg), and DIEA (12 mg) prepared in step (1.2.1) were added to a 20 ml sample vial and shaken for 16 hours. After the reaction, the reaction solution was filtered to obtain a filter cake, which was washed once with 10 ml of acetonitrile (1×10 ml) and then dried in vacuo. The dried filter cake, DMAP (3 mg), Cap1 (10 ml, 200 V), and Cap2 (1 ml, 20 V) were added to a 20 ml sample vial and shaken for 6 hours. After the reaction, the reaction solution was filtered to obtain a filter cake, which was washed once with 10 ml of acetonitrile (1×10 ml) and then dried in vacuo to obtain compound CR01008Z (1.03 g, loading 20-30 μmol / g).

[0540] Cap1 and Cap2 are capping reagents, Cap1 is a 20% by volume N-methylimidazole mixed solution in pyridine / acetonitrile, with a volume ratio of pyridine to acetonitrile of 3:5; Cap2 is a 20% by volume acetic anhydride solution in acetonitrile.

[0541] Preparation Example 2 Preparation of Compound CR01013 and Compound CR01013Z

[0542] (2.1) Preparation of compound CR01013

[0543] In this preparation example, the synthetic route of compound CR01013 is as follows:

[0544] (2.1.1) Synthesis of compound 2

[0545] Compound 1 (trans-4-(Boc-amino)cyclohexylcarboxaldehyde, 4.9 g) was dissolved in 17 ml of methanol, and aqueous formaldehyde solution (4.21 g, 37% by mass) and aqueous sodium hydroxide solution (6.5 ml, 45.3% by mass) were added dropwise. After the addition was complete, the temperature was raised to 60°C and stirred at 60°C for 2 hours. After the reaction was completed, the reaction solution was cooled to 25°C and evaporated to dryness under reduced pressure to obtain a crude white solid. A small amount of water was added to the crude product to slurry, filtered, and dried to obtain compound 2 (4.8 g, 85.9% yield) as a white solid. ESI-MS (m / z) = 260.2 [M+H] +.

[0546] (2.1.2) Synthesis of compound 3

[0547] Compound 2 (4.8 g), prepared in step (2.1.1), was dissolved in 25 ml of 1,4-dioxane. A 4 M solution of hydrochloric acid in 1,4-dioxane (25 ml) was added, and the mixture was stirred at 25°C for 2 hours. After completion of the reaction, the reaction solution was evaporated to dryness under reduced pressure to obtain compound 3 (3.6 g, 99.4% yield) as a white solid.

[0548] (2.1.3) Synthesis of compound 11

[0549] Compound 3 (3.6 g) prepared according to step (2.1.2) was dissolved in 36 ml of DMF, followed by the addition of TEA (5.62 g), compound 10 (N-benzyloxycarbonyl-4-aminobutyric acid, 5.28 g), and HBTU (8.43 g). The mixture was stirred at 25°C for 16 hours. After completion of the reaction, the reaction solution was added to 200 ml of saturated sodium bicarbonate solution and extracted three times with 100 ml of ethyl acetate (3 × 100 ml). The organic phases were combined, washed once with 50 ml of saturated sodium chloride solution (1 × 50 ml), and then dried over anhydrous sodium sulfate. The organic phase was evaporated to dryness under reduced pressure and purified by normal phase column chromatography (eluent: dichloromethane / methanol = 10 / 1, v / v) to obtain compound 11 (2.3 g, 33.0% yield) as a white solid. ESI MS (m / z) = 379.5 [M+H]+.

[0550] (2.1.4) Synthesis of compound 12

[0551] Compound 11 (2.3 g) prepared in step (2.1.3) was dissolved in 23 ml of methanol, and wet palladium on carbon (230 mg, 10% loading) was added. The atmosphere was replaced with hydrogen three times, and the reaction system was stirred at 25°C under a hydrogen atmosphere (15 psi) for 16 hours. After completion of the reaction, the reaction solution was filtered to obtain a filtrate, which was evaporated to dryness under reduced pressure to obtain compound 12 (1.48 g, 99.8% yield) as a yellow oil.

[0552] (2.1.5) Synthesis of compound 13

[0553] Compound 12 (1.48 g) prepared in step (2.1.4) was dissolved in 15 ml of DMF, and triethylamine (TEA, 1.22 g), compound 4 (1.35 g), and HBTU (3.45 g) were added. The mixture was stirred at 25°C for 16 hours. After completion of the reaction, the reaction solution was added to 150 ml of saturated sodium bicarbonate aqueous solution and extracted three times with 50 ml of ethyl acetate (3 × 50 ml). The organic phases were combined, washed once with 30 ml of saturated sodium chloride aqueous solution (1 × 30 ml), and then dried over anhydrous sodium sulfate. The organic phase was evaporated to dryness under reduced pressure and purified by reverse phase column chromatography (C18 column, eluent: water / acetonitrile = 5 / 1, v / v) to obtain compound 13 (1.3 g, yield 31.8%) as a white solid. ESI-MS (m / z): 674.3 [M+H]+.

[0554] (2.1.6) Synthesis of Compound 14

[0555] Compound 13 (1.1 g) prepared in step (2.1.5) was dissolved in 11 ml of pyridine. The reaction system was cooled to 0°C in an ice-water bath, and DMTrCl (813 mg) was added portionwise at 0°C. The reaction system was stirred at 0°C for 1 hour. After the reaction, methanol was added to quench the reaction solution. The solvent was evaporated, and the solution was purified by reverse phase column chromatography (eluent: water / acetonitrile = 1 / 4, v / v) to obtain compound 14 (800 mg, 50.3% yield) as a white solid. ESI-MS (m / z): 976.5 [M+H].

[0556] (2.1.7) Synthesis of compound CR01013

[0557] At 25°C, compound 14 (550 mg), prepared according to step (2.1.6), was dissolved in 5 ml of dichloromethane (DCM). 4,5-Dicyanoimidazole (DCl, 53.2 mg) and compound 7 (2-cyanoethyl N,N,N',N'-tetraisopropylphosphorodiamidite, 255.4 mg) were added. The atmosphere was purged with nitrogen three times, and the reaction system was stirred at 25°C under a nitrogen atmosphere for 1 hour. After completion of the reaction, the reaction solution was washed twice with 5 ml of saturated sodium bicarbonate solution (2 × 5 ml) and then once with 30 ml of saturated sodium chloride solution (1 × 30 ml). The organic phase was separated and dried over anhydrous sodium sulfate. The organic phase solvent was evaporated to dryness under reduced pressure and purified by normal phase column chromatography (eluent: dichloromethane / methanol = 20 / 1, v / v) to obtain compound CR01013 (532 mg, 80.4% yield) as a white solid. ESI-MS (m / z): 1176.7 [M+H]+.

[0558] 1H NMR(400MHz, DMSO-d6)δ0.95–1.05(d,J=6.7Hz,5H),1.06–1.15(q,J=7.6Hz,8H),1.15–1.21(t,J=7.2Hz,14H),1.72–1 .80(s,3H),1.84–1.92(s,3H),1.94–2.07(d,J=16.0Hz,7H),2.07–2.14(s,3H),2.64–2.72(q,J=5.8Hz,2H),2.74–2.89 (d,J=8.5Hz,2H),3.35–3.56(m,4H),3.57–3.70(m,4H),3.71–3.77(s,6H),3.81–3.93(m,1H),3.96–4.09(d,J=6.4Hz, 3H), 6.82–6.97 (d, J = 8.7Hz, 4H), 7.17–7.27 (t, J = 8.7Hz, 5H), 7.27–7.34 (t, J = 7.6Hz, 2H), 7.34–7.43 (d, J = 7.5Hz, 2H).

[0559] (2.2) Preparation of compound CR01013Z

[0560] In this preparation example, the synthetic route of compound CR01013Z is as follows:

[0561] (2.2.1) Synthesis of compound 15

[0562] Compound 14 (100 mg, 0.10 mmol), prepared according to step (2.1.6), was dissolved in 2 ml of dichloromethane at 25°C. Triethylamine (25.9 mg, 0.25 mmol), DMAP (1.25 mg, 0.01 mmol), and compound 8 (succinic anhydride, 15.4 mg, 0.15 mmol) were added. The reaction system was stirred at 25°C for 16 hours. After completion of the reaction, the solvent was evaporated and the product was purified by reverse phase column chromatography (C18 column, eluent: water / acetonitrile = 2 / 1, v / v) to obtain compound 15 (110 mg, 0.10 mmol, 100% yield) as a yellow oil. ESI-MS (m / z) = 1099.3 [M+Na]+.

[0563] (2.2.2) Synthesis of compound CR01013Z

[0564] Compound 15 (50 mg, 0.04 mmol), prepared according to step (2.2.1), was dissolved in 10 ml of acetonitrile. HBTU (24.2 mg, 0.06 mmol), DIEA (11.0 mg, 0.08 mmol), and amino-CPG (1.06 g, loading 80 μmol / g) were added, and the reaction system was stirred at 25°C for 16 hours. After the reaction, the reaction solution was filtered to obtain a filter cake, which was washed twice with 50 ml of dichloromethane (2 × 50 ml), three times with 50 ml of acetonitrile (3 × 50 ml), and once with 50 ml of ethyl acetate (1 × 50 ml), and then vacuum dried. Cap 1 (4.8 ml), Cap 2 (0.54 ml), and DMAP (2.59 mg) were added to the dried filter cake, and the reaction system was stirred at 25°C for 5 hours. After the reaction was completed, the reaction solution was filtered to obtain a filter cake, which was washed three times with 50 ml of acetonitrile (3×50 ml) and dried in vacuo to obtain compound CR01013Z (900 mg, loading amount of 20-30 μmol / g).

[0565] Cap1 and Cap2 are capping reagents, Cap1 is a 20% by volume N-methylimidazole mixed solution in pyridine / acetonitrile, with a volume ratio of pyridine to acetonitrile of 3:5; Cap2 is a 20% by volume acetic anhydride solution in acetonitrile.

[0566] Preparation of siRNA / siRNA conjugates

[0567] Preparation Example 3 Preparation of siRNA

[0568] (3.1) Synthetic sense strand (SS)

[0569] Using phosphoramidite solid-phase nucleic acid synthesis, nucleoside monomers are linked one by one in a 3'-5' direction. Each linking step involves four steps: deprotection, coupling, capping, and oxidation or sulfurization. The synthesis conditions are as follows:

[0570] The nucleoside monomer was prepared into an acetonitrile solution with a concentration of 0.1 M.

[0571] The deprotection reaction conditions for each step were identical: 25°C, 70 seconds, a 3% vol. dichloroacetic acid solution in dichloromethane as the deprotection reagent, and a 5:1 molar ratio of dichloroacetic acid to the 4,4'-dimethoxytrityl protecting group on the solid support.

[0572] The conditions for each coupling reaction were the same. The coupling reaction conditions were: temperature 25°C, a molar ratio of the nucleic acid sequence attached to the solid support to the nucleoside monomer of 1:10, a molar ratio of the nucleic acid sequence attached to the solid support to the coupling reagent of 1:65, a reaction time of 600 seconds, a 0.5 M solution of 5-ethylthio-1H-tetrazole in acetonitrile as the coupling reagent, and a 0.2 M solution of hydrogenated xanthan gum in acetonitrile / pyridine (a 1:1 volume ratio of acetonitrile to pyridine) as the thiolation reagent.

[0573] The capping reaction conditions were identical for each step. The capping reaction conditions were: 25°C; 2 minutes; a 1:1 molar ratio of Cap1 and Cap2; Cap1: a 20% by volume N-methylimidazole solution in pyridine / acetonitrile (3:5 volume ratio of pyridine to acetonitrile); and Cap2: a 20% by volume solution of acetic anhydride in acetonitrile. The molar ratio of the N-methylimidazole in Cap1 to the acetic anhydride in Cap2 to the nucleic acid sequence attached to the solid support was 1:1:1.

[0574] The oxidation reaction conditions were identical for each step. The oxidation reaction conditions were: temperature, 25°C; reaction time, 3 seconds; oxidizing agent concentration, 0.05 M iodine solution; a molar ratio of iodine to the nucleic acid sequence attached to the solid support during the coupling reaction, 30:1; and the oxidation reaction was performed in a water / pyridine mixture (1:9 by volume). The sulfidation reaction conditions were: temperature, 25°C; reaction time, 360 seconds; thiolation agent concentration, 0.2 M hydrogenated xanthan gum in pyridine solution; a molar ratio of thiolation agent to the nucleic acid sequence attached to the solid support during the coupling reaction, 4:1; and the sulfidation reaction was performed in a water / pyridine mixture (1:9 by volume).

[0575] After the last nucleoside monomer is connected, the nucleic acid sequence connected to the solid phase support is cut, deprotected, purified, desalted, and then freeze-dried to obtain the positive chain, wherein:

[0576] Cleavage and deprotection conditions were as follows: the synthesized nucleotide sequence attached to the solid phase support was added to 0.5 ml / μmol of 25% by mass ammonia water at 55°C for 16 hours, the solvent was removed, and the product was concentrated to dryness in vacuo. After the ammonia treatment, the product was dissolved in 0.4 ml / μmol of N-methylpyrrolidone relative to the amount of single-stranded nucleic acid, followed by the addition of 0.3 ml / μmol of triethylamine and 0.6 ml / μmol of triethylamine trihydrofluoride to remove the 2'-O-TBDMS protection from the ribose.

[0577] Purification and desalting conditions: Nucleic acid purification was achieved using a preparative ion chromatography column (Source 15Q) with a NaCl gradient elution. Specifically, eluent 1 consisted of 20 mM sodium phosphate (pH 8.1) in a water / acetonitrile mixture (9:1 volume ratio of water to acetonitrile); eluent 2 consisted of 1.5 M sodium chloride, 20 mM sodium phosphate (pH 8.1) in a water / acetonitrile mixture (9:1 volume ratio of water to acetonitrile); the elution gradient was eluent 1:eluent 2 = (100:0) to (50:50). The product eluates were collected and combined, and desalted using a reversed-phase chromatography column. Desalting conditions included using a Sephadex column with Sephadex G25 as the filler and eluting with deionized water.

[0578] Detection: Purity was determined by ion exchange chromatography (IEX-HPLC); molecular weight was determined by liquid chromatography-mass spectrometry (LC-MS). The measured molecular weight was compared with the theoretical value. If the measured value was consistent with the theoretical value, the positive strand of the siRNA was obtained.

[0579] (3.2) Synthesis of antisense strand (AS)

[0580] The antisense strand was synthesized using a universal solid phase support. The deprotection, coupling, capping, oxidation or sulfurization reaction conditions, cleavage and deprotection conditions, purification and desalting conditions in the solid phase synthesis method of the antisense strand were the same as those in step (3.1) for synthesizing the sense strand.

[0581] Detection: Purity is detected using ion exchange chromatography (IEX-HPLC); molecular weight is detected using liquid chromatography-mass spectrometry (LC-MS). The measured molecular weight is compared with the theoretical value. If the measured value is consistent with the theoretical value, it indicates that the antisense strand of the siRNA has been obtained.

[0582] (3.3) Synthesis of double-stranded siRNA

[0583] The sense chain synthesized in step (3.1) and the antisense chain synthesized in step (3.2) were mixed in an equimolar ratio, dissolved in water for injection, and heated to 95° C. The mixture was slowly cooled to room temperature and maintained at room temperature for 10 minutes to allow the sense chain and antisense chain to form a double-stranded structure through hydrogen bonds, thereby obtaining siRNA having the sense chain and antisense chain shown in Tables 4 to 7.

[0584] Table 4 siRNA sequence information

[0585] When creating RNA sequences, the sequence listing software requires that U be represented by T. Therefore, the siRNA sequences disclosed herein cannot be correctly represented in the sequence listing. All sequences are subject to those described in the specification.

[0586] Table 5 siRNA sequence information

[0587] Table 6 siRNA sequence information

[0588] Table 7 siRNA sequence information

[0589] Preparation Example 4 Preparation of siRNA Conjugate with the 3' End of the Sense Strand Conjugated to the L96 Vector

[0590] Compound L96-PS was purchased from Ailiying Pharmaceutical Group (Tianjin) Co., Ltd. with a loading of 120±12 μmol / g (detection method: UV / HPLC). The structural formula of compound L96-PS is as follows:

[0591] Wherein, PS represents polystyrene resin solid phase carrier.

[0592] (4.1) Synthesis of the positive chain:

[0593] Using the phosphoramidite nucleic acid solid-phase synthesis method, the L96-PS compound attached to the solid support was used as the initial cycle, and nucleoside monomers were attached one by one in the 3'-5' direction according to the nucleotide sequence. Each nucleoside monomer attachment involved a four-step reaction: deprotection, coupling, capping, and oxidation or sulfurization. The synthesis method and conditions were similar to the sense strand synthesis method described in step (3.1) of Preparation Example 3.

[0594] (4.2) Synthesis of antisense strand:

[0595] The siRNA antisense strand in this preparation example was synthesized according to the antisense strand synthesis method shown in step (3.2) of Preparation Example 3.

[0596] (4.3) Synthesis of siRNA conjugates

[0597] The siRNA conjugate of this preparation example was synthesized according to the method shown in step (3.3) of Preparation Example 3.

[0598] The structural formula of the siRNA conjugate in which the 3' end of the sense chain is conjugated to the L96 vector is as follows:

[0599] in, L96 is conjugated to the 3' end of the sense strand of the siRNA via a phosphodiester bond.

[0600] Table 8 Sequence information of siRNA conjugates with the 3' end of the sense strand conjugated to the L96 vector

[0601] Exemplarily explained, "SEQ ID No. 849_L96" means that the L96 vector is conjugated to the 3' end of the sense strand as shown in SEQ ID No. 849 via a phosphodiester bond.

[0602] Table 9 Sequence information of mouse siRNA conjugates with the 3' end of the sense strand conjugated to the L96 vector

[0603] Table 10 Sequence information of siRNA conjugates with the 3' end of the positive strand conjugated to the L96 vector

[0604] Preparation Example 5 Synthesis of siRNA Conjugate Conjugated to the 3' End of the Sense Strand (CR01008×3) Vector

[0605] (5.1) Synthesis of the positive chain

[0606] By the phosphoramidite nucleic acid solid phase synthesis method, starting with the compound CR01008Z connected to the solid phase support, nucleoside monomers are linked one by one in a 3'-5' direction according to the nucleotide sequence (compound CR01008 is regarded as one nucleoside monomer).

[0607] Each linking of a nucleoside monomer involves four steps: deprotection, coupling, capping, and oxidation or sulfurization. The deprotection, coupling, capping, oxidation or sulfurization reaction conditions, cleavage and deprotection conditions, and purification and desalting conditions in the synthesis of the sense chain in this preparation example are the same as those in the synthesis of the sense chain in step (3.1) of Preparation Example 3.

[0608] In this step, three clusters of CR01008 were synthesized during the synthesis of the sense strand, which were denoted as (CR01008)×3 or (CR01008×3).

[0609] The structural formula of the three-cluster CR01008 is as follows:

[0610] (5.2) Synthesis of antisense strand:

[0611] The antisense strand of this preparation example was synthesized according to the antisense strand synthesis method shown in step (3.2) of Preparation Example 3.

[0612] (5.3) Synthesis of siRNA conjugates

[0613] The siRNA conjugate of this preparation example was synthesized according to the method shown in step (3.3) of Preparation Example 3.

[0614] When the ligand is a triple cluster of CR01008, the structural formula of the siRNA conjugate is as follows:

[0615] in, Represents siRNA. (CR01018×3) The vector is conjugated to the 3' end of the sense strand of the siRNA.

[0616] Table 11 Sequence information of siRNA conjugates with the 3' end of the positive strand conjugated to a vector (CR01008×3)

[0617] For illustrative explanation of the above table, "SEQ ID No. 858_(CR01008×3)" indicates that the (CR01008×3) vector is conjugated to the 3' end of the positive strand shown in SEQ ID No. 858.

[0618] Biological detection experiments

[0619] Example 1 Evaluation of the in vitro activity of Complement C3 (C3) siRNA

[0620] In this example, the target gene inhibitory activity evaluation method in the liver cancer cell line HepG2 was used to evaluate the inhibitory activity of RX002001 to RX002212 at the same C3 target site on the target gene C3 mRNA (accession number NM_000064.4) in cells.

[0621] Test sample preparation:

[0622] After centrifugation, dissolve the siRNA sample in DEPC H2O according to the specifications of each tube to prepare a 20 μM stock solution. This stock solution was then serially diluted with DEPC H2O to produce 0.04 μM and 0.004 μM working solutions. Dose screening experiments were performed at final duplex concentrations of 1 nM and 0.1 nM.

[0623] 96-well transfection and detection:

[0624] HepG2 cells grown to near confluence were trypsinized, washed, and a cell suspension was prepared for plating in 96-well plates.

[0625] Disperse 5 μL of 0.4 μM and 0.04 μM working solutions in 20 μL of Opti-MEM to form an siRNA mixture. Disperse 0.2 μL of RNAiMAX in 24.8 μL of Opti-MEM and incubate for 5 minutes. Then, mix with each siRNA mixture and incubate for 10 minutes to form a transfection complex. Add 150 μL of cell suspension (12,000 cells) per well to the transfection complex, and place the 96-well plate in an incubator for an additional 24 hours.

[0626] For the mock control group, 0.2 μL RNAiMAX was dispersed in 24.8 μL Opti-MEM and incubated for 5 minutes. Then, 25 μL Opti-MEM was added and incubated for 10 minutes to form a transfection complex. 150 μL of cell suspension (12,000 cells) was added to the transfection complex per well, and the 96-well plate was placed in an incubator and incubated for 24 hours.

[0627] Remove the 96-well plate, lyse the cells, and dilute the lysate. Use a one-step qPCR kit (Thermo Fisher, Power SYBR Green RNA-to-CT TM 1-Step, 4389986) Target gene mRNA expression in HeLa cells was detected on a fluorescent quantitative PCR instrument (Roche, LightCycler 480). In this real-time fluorescent quantitative PCR method, the TATA-box binding protein (TBP) gene was used as an internal reference gene. Primers targeting the target gene and primers targeting the TBP internal reference gene were used to detect the target gene and TBP internal reference gene, respectively. The sequences of the detection primers are shown in Table 12.

[0628] Table 12 Sequences of detection primers

[0629] In the real-time fluorescence quantitative PCR method, the expression level and inhibition rate of the target gene mRNA in each test group were relatively quantitatively calculated using the ΔΔCt method according to the technical method described in the embodiment.

[0630] Table 13 Inhibitory activity of target genes in HepG2 cells after administration of the siRNA described in this example

[0631] Example 2 Evaluation of the activity of siRNA conjugates with the 3' end of the sense strand conjugated to the L96 vector in BALB / c-HDI mice

[0632] In this example, the BALB / c mouse tail vein hydrodynamic injection model was used to evaluate the inhibitory activity of the same C3 target siRNA sense strand 3' end conjugated to the L96 carrier conjugates RZ002001-RZ0020032 on the target gene C3 mRNA, with RZ000001 as a negative control.

[0633] Plasmid construction:

[0634] pcDNA-CMV-RG002 plasmid (ID: NM_000064.4) was constructed by Sangon Biotech (Shanghai) Co., Ltd.

[0635] Mouse model construction:

[0636] The BALB / c mouse hydrodynamic injection model is established by rapidly injecting a pcDNA-CMV-RG002 plasmid solution through the tail vein at high pressure. On day 3 of the experiment, mice were injected with 10 μg of pcDNA-CMV-RG002 via the tail vein hydrodynamic injection over 5 seconds, at an injection volume of 8% of their body weight. The plasmid DNA for injection was diluted in PBS and the solution was prepared immediately prior to injection and stored at 4°C.

[0637] Animal grouping, drug administration and tissue sample collection:

[0638] 6-7 week old BALB / c mice were randomly divided into groups (all female) by body weight, with 5 mice in each group. Each test group was given a predetermined dose of drug conjugate and a PBS control group was added. All mice were dosed according to body weight, and a single dose was administered by subcutaneous injection in the abdomen. Each drug conjugate was administered in the form of a PBS solution of 0.1 mg (calculated as siRNA) / mL, and the administration volume was 10 mL / kg (mouse body weight). That is, the dosage of each drug conjugate was 1 mg (calculated as siRNA) / kg (mouse body weight). The PBS control group was given 10 mL / kg (mouse body weight) of PBS solution without siRNA conjugate. The administration day was recorded as day 0 (recorded as D0), and plasmid injection was performed on the 3rd day after administration (recorded as D3). Five mice of all groups were killed on the 4th day (recorded as D4). The mice were subjected to gross dissection and the liver tissue of each mouse was collected. The liver tissue was cut into about 2 mm 3 Small pieces were stored with RNA later.

[0639] For each mouse, an appropriate amount of liver tissue sample was taken from RNA later, and the liver tissue sample was disrupted in a Tissuelyser II fully automatic tissue homogenizer for 60 seconds. Then, total RNA was extracted using a fully automatic nucleic acid extractor (purchased from Zhejiang Hanwei Technology Co., Ltd.) and a nucleic acid extraction kit (purchased from Zhejiang Hanwei Technology Co., Ltd., GO-MNTR-100) according to the standard operating procedures for total RNA extraction.

[0640] For each mouse, 1 μg of total RNA was taken respectively, and a reverse transcription kit (Thermo Fisher Scientific, RevertAid First Strand cDNA Synthesis Kit, K1622) was used and Oligo (dT) 18 reverse transcription primer was selected. According to the method described in the reverse transcription kit instructions, 20 μL reverse transcription system was configured and the reverse transcription reaction was completed. After the reaction was completed, 60 μL RNase-Free water was added to the reverse transcription system to obtain a cDNA solution. The expression level of the target gene mRNA in the animal was then detected using a real-time fluorescence quantitative PCR kit (Thermo Fisher Scientific, TaqMan Fast Advanced Master Mix, 4444557) and a fluorescence quantitative PCR instrument (Bio-Rad, CFX Opus 384). In this real-time fluorescence quantitative PCR method, the Nero gene on the plasmid backbone was used as an internal reference gene, and the target gene and the Nero internal reference gene were detected using primers for the target gene and primers for the Nero internal reference gene, respectively. The sequences of the detection primers are shown in Table 14.

[0641] Table 14 Sequences of detection primers

[0642] According to the instructions of the real-time fluorescence quantitative PCR kit, 10 μL of real-time PCR reaction system was prepared for each PCR detection well. Each reaction system contained 4 μL of the cDNA solution obtained by the reverse transcription reaction, 5 μL of TaqMan TMFast Advanced Master Mix (2×), 0.15 μL 10 μM upstream primer, 0.15 μL 10 μM downstream primer (primer information see Table 8), 0.15 μL 10 μM probe primer, 0.55 μL RNase-Free H2O. The configured reaction system was placed on a real-time fluorescence quantitative PCR instrument (Bio-Rad, CFX Opus 384), and Real-time PCR amplification was performed using a two-step method. The amplification program was 50°C for 2 min, followed by pre-denaturation at 95°C for 20 s, denaturation at 95°C for 3 s, annealing and extension at 60°C for 30 s, and the denaturation, annealing and extension process was repeated for 40 cycles. In this real-time fluorescence quantitative PCR method, the expression level and inhibition rate of the target gene mRNA in each test group were relatively quantitatively calculated using the ΔΔCt method according to the technical method described in the embodiment.

[0643] The results of Example 2 showed that at a dose of 1 mg / kg, RZ002003, RZ002006, RZ002011, RZ002015 and RZ002031 could significantly inhibit the expression of C3 gene mRNA in mouse liver tissue ( FIG. 1 , Table 15 ).

[0644] Table 15 Inhibitory activity of C3 gene mRNA in liver tissue of BALB / c-HDI mice after administration of siRNA conjugates described in this example

[0645] Example 3 Toxicological evaluation of siRNA conjugates with the 3' end of the sense strand conjugated to the L96 vector in ICR mice

[0646] In this example, a single subcutaneous injection of 300 mg / kg was used to evaluate the nature and extent of the toxic reactions of the L96 vector conjugates RZ002003, RZ002006, RZ002011, RZ002015, and RZ002031 in ICR mice.

[0647] Animal grouping, drug administration and tissue sample collection:

[0648] 6-8 week old ICR mice (purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.) were randomly divided into groups according to body weight, with 10 mice per group, half male and half female. Each test group was given a predetermined dose of drug conjugate and a PBS control group was added. All mice were dosed according to body weight and administered by subcutaneous injection in the abdomen as a single dose. Each drug conjugate was administered in the form of a 30 mg (calculated as siRNA) / mL PBS solution, and the administration volume was 10 mL / kg (mouse body weight). That is, the dosage of each drug conjugate was 300 mg (calculated as siRNA) / kg (mouse body weight). The PBS control group was given 10 mL / kg (mouse body weight) of PBS solution without siRNA conjugate and observed for 14 days. The day of administration was recorded as day 0 (denoted as D0). Clinical observations were performed at least once a day after the first administration during the experimental period, and body weight and food intake were measured twice a week. On the 14th day of the experiment (denoted as D14), blood was drawn from all animals (mice were fasted for at least 12 hours before sampling and testing, and water was not allowed) for hematology and blood biochemistry tests. Blood was collected and anticoagulated with EDTA-K2, and routine blood tests were performed using an automated modular blood and body fluid analyzer (Sysmex XN-10B1). Blood samples (without anticoagulation) were collected and placed at room temperature for approximately 30 minutes. After blood coagulation, they were centrifuged at 2000 g for 10 minutes at 4°C, and blood biochemical analysis was performed using a fully automated biochemical analyzer (Sysmex BX-4000). All animals were dissected on day 14 of the experiment (designated D14). Mice were fasted for at least 12 hours prior to dissection and anesthetized with Zotal 50 (Vicco, 8G4VA, France). After blood collection, the animals were euthanized by exsanguination of the abdominal aorta for gross anatomical observation. Histopathological examinations of the liver and kidneys of all animals were performed, as shown in Figure 2 (stained with hematoxylin-eosin). The results showed that none of the test substances had significant effects on the general condition, body weight, and gross anatomical observations of ICR mice. Compared with the PBS group, at a dose of 300 mg (calculated as siRNA) / kg (mouse body weight), no significant abnormalities were observed in the hematology and blood biochemistry of animals in the RZ002003, RZ002006, RZ002011, and RZ002015 groups; no significant abnormalities were observed in the hematology of animals in the RZ002031 group, with slight increases in ALT and AST.

[0649] Example 4 In vitro activity evaluation of siRNA conjugates containing the 3' end of the sense strand conjugated to (CR01008) × 3 vectors

[0650] In this example, the target gene inhibitory activity evaluation method in the human liver cancer cell line HepG2 was used to evaluate the inhibitory activity of the (CR01008)×3 vector siRNA conjugate on the target gene C3 mRNA in cells.

[0651] Test sample preparation:

[0652] After centrifugation, dissolve each siRNA sample in PBS according to the specifications of each tube to prepare a 20 μM stock solution. This stock solution was then serially diluted with PBS to produce 0.5 μM and 0.05 μM working solutions. Dosage testing was performed at final duplex concentrations of 5 nM and 0.5 nM.

[0653] 96-well transfection and detection:

[0654] HepG2 cells grown to near confluence were digested with trypsin, and the cells were washed to prepare a cell suspension. 100 μL of cell suspension was added to each well of a 96-well plate, with 12,000 cells per well. The cells were cultured in a 37°C, 5% CO2 incubator. After the cells adhered for 24 hours, the DMEM medium in the 96-well plate was aspirated and 80 μL of Opti-MEM was added to each well. TM Culture medium was added, and the 96-well plate was then placed in an incubator for further incubation. 1 μL of the 0.1 μM and 0.01 μM working solutions were dispersed in 9 μL of Opti-MEM to form the siRNA mixture. 0.3 μL of RNAiMAX was dispersed in 9.7 μL of Opti-MEM and mixed with each siRNA mixture to form a transfection complex. The transfection complex was incubated at room temperature for 10 minutes, and then the transfection complex was added to the 96-well plate at a rate of 20 μL / well. After 4 hours of incubation, 100 μL of DMEM medium containing 20% ​​FBS was added to each well, and the 96-well plate was placed in an incubator for a further 24 hours. For the mock control group, 0.3 μL of RNAiMAX was dispersed in 9.7 μL of Opti-MEM, followed by 10 μL of Opti-MEM. The mixture was incubated at room temperature for 10 minutes; then, 20 μL of the complex was added to the 96-well plate at a rate of 20 μL / well. After culturing for 4 hours, 100 μL of DMEM medium containing 20% ​​FBS was added to each well, and the 96-well plate was placed in an incubator and cultured for a further 24 hours.

[0655] The 96-well plate was removed and total RNA was extracted using a fully automatic nucleic acid extractor (purchased from Zhejiang Hanwei Technology Co., Ltd.) and a nucleic acid extraction kit (purchased from Zhejiang Hanwei Technology Co., Ltd., GO-MNTR-100) according to the standard operating procedures for total RNA extraction.

[0656] Use reverse transcription kit (Thermo Fisher Scientific company, RevertAid First Strand cDNA Synthesis Kit, K1622) and select Oligo (dT) 18 reverse transcription primer, configure 20 μ L reverse transcription system according to the method for reverse transcription kit specification sheet record and complete reverse transcription reaction.Then use real-time fluorescence quantitative PCR kit (Thermo Fisher Scientific company, TaqMan Fast Advanced Master Mix, 4444557) to detect the expression of target gene mRNA in HepG2 cells on fluorescence quantitative PCR instrument (Bio-Rad company, CFX Opus384).In this real-time fluorescence quantitative PCR method, using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene as internal reference gene, use the primer for target gene and the primer for GAPDH internal reference gene to detect target gene and GAPDH internal reference gene respectively.The sequence of detection primer is shown in Table 16.

[0657] Table 16 Sequences of detection primers

[0658] According to the instructions of the real-time fluorescence quantitative PCR kit, 10 μL of real-time PCR reaction system was prepared for each PCR detection well. Each reaction system contained 4 μL of the cDNA solution obtained by the reverse transcription reaction, 5 μL of TaqMan TM Fast Advanced Master Mix (2×), 0.15 μL of 10 μM upstream primer, 0.15 μL of 10 μM downstream primer (primer information see Table 14), 0.15 μL of 10 μM probe primer, and 0.55 μL of RNase-Free H2O. The prepared reaction system was placed in a real-time fluorescence quantitative PCR instrument (Bio-Rad, CFX Opus 384) and real-time PCR amplification was performed using a two-step method: 50°C for 2 min, followed by pre-denaturation at 95°C for 20 s, denaturation at 95°C for 3 s, annealing and extension at 60°C for 30 s, and 40 cycles of denaturation, annealing, and extension. In this real-time fluorescence quantitative PCR method, the expression level and inhibition rate of the target gene C3 mRNA in each test group were relatively quantitatively calculated using the ΔΔCt method according to the technical methods described in the embodiment (Figure 3).

[0659] Table 17 Inhibitory activity of target gene C3 mRNA in HepG2 cells after administration of the siRNA conjugates described in this example

[0660] The results of Example 4 showed that at doses of 5 nM and 0.5 nM, the differently modified CR01008 vector conjugate siRNAs could significantly inhibit the expression of C3 gene mRNA in HepG2 cells (Table 17).

[0661] Example 5 Evaluation of mRNA-lowering effect of siRNA conjugates containing the 3' end of the sense strand conjugated to (CR01008) × 3 vectors in normal cynomolgus monkeys

[0662] This example evaluates the inhibitory activity of RZ002099, RZ002101, RZ002106, and RZ002113 on the target gene C3 mRNA in cynomolgus monkeys.

[0663] Animal grouping, drug administration and tissue sample collection:

[0664] Healthy cynomolgus monkeys weighing 3-5 kg ​​were divided into groups of 4 per group, half male and half female. Each test group was given a predetermined dose of drug conjugate and a PBS control group was added. The dosage of all animals was calculated according to body weight and a single dose was given by subcutaneous injection in the abdomen. Each drug conjugate was administered in the form of a PBS solution of 9 mg (calculated as siRNA) / mL, and the administration volume was 1 mL / kg (cynomolgus monkey body weight), that is, the dosage of each drug conjugate was 9 mg (calculated as siRNA) / kg (cynomolgus monkey body weight). The PBS control group was given 1 mL / kg (cynomolgus monkey body weight) of PBS solution without siRNA conjugate. The day of administration was recorded as day 0 (recorded as D0), and on the 14th day after administration (recorded as D14), each group of cynomolgus monkeys underwent liver biopsy, and the liver tissue was immediately preserved with RNA later.

[0665] For each crab-eating macaque, liver tissue samples were removed from RNA later and disrupted in a Tissuelyser II fully automatic tissue homogenizer for 60 seconds. Total RNA was then extracted using a fully automatic nucleic acid extractor (purchased from Zhejiang Hanwei Technology Co., Ltd.) and a nucleic acid extraction kit (purchased from Zhejiang Hanwei Technology Co., Ltd., GO-MNTR-100) according to the standard operating procedures for total RNA extraction.

[0666] For each cynomolgus monkey, 1 μg of total RNA was taken respectively, and a reverse transcription kit (Thermo Fisher Scientific, RevertAid First Strand cDNA Synthesis Kit, K1622) was used and Oligo (dT) 18 reverse transcription primer was selected. According to the method described in the reverse transcription kit instructions, 20 μL reverse transcription system was configured and the reverse transcription reaction was completed. After the reaction was completed, 60 μL RNase-Free water was added to the reverse transcription system to obtain a cDNA solution. Then, a real-time fluorescence quantitative PCR kit (Thermo Fisher Scientific, TaqMan Fast Advanced Master Mix, 4444557) was used to detect the expression of the target gene mRNA in the animal on a fluorescence quantitative PCR instrument (Roche, Lightcycler 480II). In this real-time fluorescence quantitative PCR method, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as an internal reference gene, and the target gene and the GAPDH internal reference gene were detected using primers for the target gene and primers for the GAPDH internal reference gene, respectively. The sequences of the detection primers are shown in Table 18.

[0667] Table 18 Sequences of detection primers

[0668] According to the instructions of the real-time fluorescence quantitative PCR kit, 10 μL of real-time PCR reaction system was prepared for each PCR detection well. Each reaction system contained 4 μL of the cDNA solution obtained by the reverse transcription reaction, 5 μL of TaqMan TM Fast Advanced Master Mix (2×), 0.15 μL 10 μM upstream primer, 0.15 μL 10 μM downstream primer (primer information see Table 16), 0.15 μL 10 μM probe primer, 0.55 μL RNase-Free H2O. The configured reaction system was placed on a real-time fluorescence quantitative PCR instrument (Bio-Rad, CFX Opus 384), and Real-time PCR amplification was performed using a two-step method. The amplification program was 50°C for 2 min, followed by pre-denaturation at 95°C for 20 s, denaturation at 95°C for 3 s, annealing and extension at 60°C for 30 s, and the denaturation, annealing and extension process was repeated for 40 cycles. In this real-time fluorescence quantitative PCR method, the expression level and inhibition rate of the target gene mRNA in each test group were relatively quantitatively calculated using the ΔΔCt method according to the technical method described in the embodiment.

[0669] The results of Example 5 showed that RZ002099, RZ002101, RZ002106 and RZ002113 could significantly inhibit the expression of C3 gene mRNA at a single dose of 9 mg / kg (Table 19, Figure 4).

[0670] Table 19: Changes in C3 mRNA levels in the liver of cynomolgus monkeys after administration of the siRNA conjugates described in this example

[0671] Example 6 Evaluation of the C3 protein-lowering effect of the siRNA conjugate containing the 3' end of the sense strand conjugated to (CR01008) × 3 vectors in normal cynomolgus monkeys

[0672] In this example, the ELISA method was used to determine the expression of C3 protein in the serum of cynomolgus monkeys at different time points after a single administration of (CR01008)×3 carrier conjugates RZ002099, RZ002101, RZ002106 and RZ002113.

[0673] Animal grouping, drug administration and tissue sample collection:

[0674] Healthy cynomolgus monkeys weighing 3-5 kg ​​were grouped according to serum C3 protein levels, with 4 animals per group, half male and half female. Each test group was given a predetermined dose of the drug conjugate and a PBS control group was added. All animals were dosed according to body weight and administered as a single subcutaneous injection in the abdomen. Each drug conjugate was administered in the form of a PBS solution of 9 mg (calculated as siRNA) / mL, with a dosing volume of 1 mL / kg (cynomolgus monkey body weight), i.e., the dose of each drug conjugate was 9 mg (calculated as siRNA) / kg (cynomolgus monkey body weight). The PBS control group was given 1 mL / kg (mouse body weight) of PBS solution without siRNA conjugate. Serum was collected from cynomolgus monkeys before administration (denoted as pre-dose value, pre-dose), on the day of administration as day 0 (denoted as D0), on the 7th day (denoted as D7), the 14th day (denoted as D14), the 21st day (denoted as D21), the 28th day (denoted as D28), the 35th day (denoted as D35), the 42nd day (denoted as D42), the 49th day (denoted as D49), the 56th day (denoted as D56), the 63rd day (denoted as D63), the 70th day (denoted as D70), the 77th day (denoted as D77), the 84th day (denoted as D84), the 91st day (denoted as D91), the 98th day (denoted as D98), and the 105th day (denoted as D105) after administration, and the C3 protein expression was determined using a human complement C3 ELISA kit (HycultBiotech, HK366-01).

[0675] The results of Example 6 showed that, at a single 9 mg / kg dose, RZ002099, RZ002101, RZ002106, and RZ002113 all significantly reduced C3 protein levels in cynomolgus monkey serum. RZ002106 achieved a 90% reduction in C3 protein, with an 80% reduction observed at D70. The levels had not returned to pre-drug levels by D105 ( Figure 5 , Table 20 ).

[0676] Table 20 Changes in C3 protein levels in cynomolgus monkey serum after administration of the siRNA conjugates described in this example

[0677] Example 7 Effect of siRNA conjugates containing the 3' end of the sense strand conjugated to (CR01008) × 3 vectors on the activity of the complement pathway in normal cynomolgus monkeys

[0678] This example evaluates the complement activity in cynomolgus monkey serum at different time points after a single administration of the (CR01008)×3 vector (i.e., (CR01008×3) vector) conjugates RZ002099, RZ002101, RZ002106, and RZ002113.

[0679] Animal grouping, drug administration and tissue sample collection:

[0680] Healthy cynomolgus monkeys weighing 3-5 kg ​​were divided into groups of 4, half male and half female. Each test group was given a predetermined dose of the drug conjugate and a PBS control group was added. All animals were dosed according to body weight and administered as a single subcutaneous injection in the abdomen. Each drug conjugate was administered as a 9 mg (calculated as siRNA) / mL PBS solution in a dosing volume of 1 mL / kg (cynomolgus monkey body weight), i.e., the dose of each drug conjugate was 9 mg (calculated as siRNA) / kg (cynomolgus monkey body weight). The PBS control group was given 1 mL / kg (cynomolgus monkey body weight) of PBS solution without the siRNA conjugate. Before administration (recorded as pre-dose value, pre-dose), the day of administration was recorded as day 0 (recorded as D0), and the 7th day (recorded as D7), the 14th day (recorded as D14), the 21st day (recorded as D21), the 28th day (recorded as D28), the 35th day (recorded as D35), the 42nd day (recorded as D42), the 49th day (recorded as D49), the 56th day (recorded as D56), the 63rd day (recorded as D63), the 70th day (recorded as D70), the 77th day (recorded as D77), the 84th day (recorded as D84), the 91st day (recorded as D91), the 98th day (recorded as D98), and the 105th day (recorded as D105) were used. The activities of the complement system alternative pathway (CAP) and complement system classical pathway (CCP) were determined using the complement system alternative pathway (COMPL AP330 RUO, IBL America) and complement system classical pathway kits (COMPL CP310 RUO, IBL America).

[0681] The results of Example 7 showed that at a single dose of 9 mg / kg, RZ002099, RZ002101, RZ002106 and RZ002113 could significantly inhibit the alternative complement pathway in cynomolgus monkey serum, but had no significant effect on the classical complement pathway in serum ( Figure 6 , Figure 7 , Table 21 , Table 22 ).

[0682] Table 21 Changes in serum CAP activity in cynomolgus monkeys after administration of the siRNA conjugates described in this example

[0683] Table 22 Changes in serum CCP activity in cynomolgus monkeys after administration of the siRNA conjugates described in this example

[0684] Example 8: Evaluation of the efficacy of siRNA conjugates with the 3' end of the sense strand conjugated to the L96 vector in CFA-hIgA mice

[0685] In this example, a mouse model of IgA nephropathy was established using complete Freund's adjuvant (CFA)-induced humanized IgA mice. The inhibitory effect of RZM02003 on mouse C3 mRNA expression in CFA-IgA mice was assessed using quantitative polymerase chain reaction (QPCR). Serum mouse C3 protein expression at different time points was measured using an enzyme-linked immunosorbent assay (ELISA). Immunofluorescence was used to observe the deposition of mouse C3 protein and human IgA immune protein in renal tissue. Periodic acid-Schiff (PAS) staining was used to observe renal tissue lesions.

[0686] Animal grouping, drug administration and tissue sample collection:

[0687] CFA-hIgA model mice were randomly divided into groups of 3 mice per group. Each test group was given a predetermined dose of the drug conjugate and a PBS control group was added. The drug dose for all mice was calculated based on body weight, and the administration volume was 10 mL / kg (mouse body weight). RZM02003 was administered subcutaneously in the abdomen as a single dose, with the drug conjugate administered in the form of 0.9 mg (calculated as siRNA) / mL of PBS solution, i.e., the drug conjugate dosage was 9 mg (calculated as siRNA) / kg (rat body weight). The PBS control group was given 10 mL / kg (mouse body weight) of PBS solution without the siRNA conjugate.

[0688] Before administration, the renal apex tissue of each mouse after modeling was collected by surgical excision. A portion of the renal tissue was fixed with 10% formalin fixative, embedded in paraffin, and sliced ​​and then PAS stained to observe renal tissue lesions before administration; a portion of the liver tissue was embedded and fixed with OCT (optimal cutting temperature compound), and frozen sliced ​​and then immunofluorescence staining of the kidney tissue was performed to observe mouse C3 deposition and human IgA deposition in the kidney before administration. The day of administration was recorded as day 0 (recorded as D0). Three mice in all groups were killed on the 14th day after administration (recorded as D14), and three mice in all groups were killed on the 28th day after administration (recorded as D28). The serum of all sacrificed mice was collected, and the expression of C3 protein in the serum was detected using the mouse complement C3 ELISA kit (abcam, 157711). The sacrificed mice were subjected to gross dissection, and the liver tissue of each sacrificed mouse was collected. The liver tissue was cut into approximately 2mm 3 Small pieces were collected and stored in RNA later. Kidney tissue was collected from each sacrificed mouse. A portion of the kidney tissue was fixed with 10% formalin, embedded in paraffin, and sectioned for PAS staining to observe renal tissue lesions on days 14 and 28 after administration. A portion of the liver tissue was embedded in OCT, fixed, and frozen sectioned for immunofluorescence staining to observe mouse C3 and human IgA deposition in the kidneys on days 14 and 28 after administration.

[0689] The results of Example 8 showed that a single dose of 9 mg / kg of RZM02003 significantly reduced C3 mRNA and protein levels in CFA-IgA mice. On day 14 after administration, the C3 mRNA and protein reduction effects reached 98% ( Figure 8 , Table 23 ) and 78% ( Figure 9 , Table 24 ), respectively. On day 28 after administration, the C3 mRNA reduction effect was still greater than 98% ( Figure 8 , Table 23 ), and the C3 protein reduction effect was approximately 70% ( Figure 9 , Table 24 ).

[0690] The results of Example 8 showed that RZM02003 could reduce C3 deposition in renal tissue at a single dose of 9 mg / kg ( FIG. 10 ), and a significant decrease in C3 fluorescence staining in renal tissue was observed on days 14 and 28 after administration.

[0691] Table 23 Inhibitory activity of C3 mRNA in the liver of CFA-hIgA mice after administration of the siRNA conjugates described in this example

[0692] Table 24: Changes in C3 protein levels in the serum of CFA-hIgA mice after administration of the siRNA conjugates described in this example

[0693] The double-stranded oligonucleotides and conjugates for inhibiting C3 gene expression provided herein, as well as their uses, have been described in detail. Specific examples have been used herein to illustrate the principles and implementation methods of the present disclosure. The descriptions of the above examples are intended only to facilitate understanding of the methods and core concepts of the present disclosure. It should be noted that those skilled in the art may make various improvements and modifications to the present disclosure without departing from the principles of the present disclosure, and such improvements and modifications are also within the scope of protection of the claims of the present disclosure.

Claims

1. A double-stranded oligonucleotide for inhibiting C3 gene expression, characterized in that: The double-stranded oligonucleotide comprises a sense strand and an antisense strand; In the direction from the 5' end to the 3' end, nucleotides 2 to 19 of the antisense strand comprise a complementary region to a C3RNA transcript, wherein the complementary region comprises at least 17 consecutive nucleotides in any one of SEQ ID Nos. 213 to 424, or a nucleotide sequence having 1, 2 or 3 nucleotide differences with the consecutive nucleotides; The sense strand contains at least 17 nucleotides, and the sense strand and the antisense strand are complementary or substantially complementary to form a duplex region; the substantially complementary means that the mismatch between the sense strand and the antisense strand in the duplex region does not exceed 3 nucleotides.

2. The double-stranded oligonucleotide according to claim 1, characterized in that The antisense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 213 to 424 and has 0, 1, 2 or 3 nucleotide differences; Optionally, the antisense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 213 to 424 and having 0 or 1 nucleotide difference; Optionally, the antisense strand is selected from or comprises any one of the nucleotide sequences of SEQ ID No. 213 to 424.

3. The double-stranded oligonucleotide according to claim 1, characterized in that The sense strand comprises at least 17 consecutive nucleotides in any one of the nucleotide sequences of SEQ ID NOs. 1 to 212, or a nucleotide sequence having 1, 2 or 3 nucleotide differences from the consecutive nucleotides; Optionally, the sense strand comprises a nucleotide sequence of any one of SEQ ID NOs. 1 to 212 and having 0, 1, 2 or 3 nucleotide differences; Optionally, the sense strand comprises a nucleotide sequence of any one of SEQ ID NOs. 1 to 212 and having 0 or 1 nucleotide difference; Optionally, the sense strand is selected from or comprises any one of the nucleotide sequences of SEQ ID NOs. 1 to 212.

4. The double-stranded oligonucleotide according to claim 3, characterized in that The antisense strand comprises a nucleotide sequence of any one of SEQ ID NO. 241, 242, 285, 357 or 403 and has 0 or 1 nucleotide difference; Optionally, the antisense strand comprises any one of the nucleotide sequences of SEQ ID NOs. 241, 242, 285, 357 and 403; Optionally, the sense strand comprises a nucleotide sequence of any one of SEQ ID NOs. 29, 30, 73, 145 and 191 and has 0 or 1 nucleotide difference; Optionally, the sense strand comprises any one of the nucleotide sequences of SEQ ID NOs. 29, 30, 73, 145 and 191; Optionally, the double-stranded oligonucleotide comprises one or more of the following: An antisense strand having a nucleotide sequence as shown in SEQ ID NO.241 or having 0 or 1 nucleotide difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID NO.29 or having 0 or 1 nucleotide difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID NO.242 or having 0 or 1 nucleotide difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID NO.30 or having 0 or 1 nucleotide difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID NO.285 or having 0 or 1 nucleotide difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID NO.73 or having 0 or 1 nucleotide difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID NO.357 or having 0 or 1 nucleotide difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID NO.145 or having 0 or 1 nucleotide difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID NO.403 or having 0 or 1 nucleotide difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID NO.191 or having 0 or 1 nucleotide difference therefrom; Optionally, the double-stranded oligonucleotide comprises: An antisense strand having a nucleotide sequence as shown in SEQ ID NO.241 or having 0 or 1 nucleotide difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID NO.29 or having 0 or 1 nucleotide difference therefrom.

5. The double-stranded oligonucleotide according to any one of claims 1 to 4, characterized in that Each nucleotide in the double-stranded oligonucleotide is independently selected from modified or unmodified nucleotides; Optionally, substantially all nucleotides in the double-stranded oligonucleotide are independently selected from modified nucleotides; Optionally, all nucleotides in the double-stranded oligonucleotide are independently selected from modified nucleotides; Optionally, each nucleotide in the double-stranded oligonucleotide is independently selected from the following modified nucleotides: 2'-halogenated modified nucleotides, 2'-deoxy modified nucleotides, 2'-O-optionally substituted C 1-6 Alkyl-modified nucleotides, 2'-O-(CH2) n -O-R1 modified nucleotides, 2'-amino modified nucleotides, abasic nucleotides, or quasi-nucleotides; the quasi-nucleotides are selected from one or more of PNA, MNA, BNA, LNA, GNA, TNA or UNA; wherein n is selected from 1 or 2, R1 is selected from optionally substituted C 1-6 Alkyl or optionally substituted C 1-6 Alkoxy; if R1 contains a substituent, the substituent is selected from halogen, C 1-6 Alkoxy, hydroxy or amino; Optionally, the 2'-halogenated modified nucleotides are selected from 2'-fluoro-modified nucleotides; Optionally, the 2'-O-optionally substituted C 1-6 The alkyl-modified nucleotides are selected from 2'-O-methyl-modified nucleotides; Optionally, the 2'-O-(CH2) n -O-R1 modified nucleotide is selected from 2'-O-methoxyethyl modified nucleotide, 2'-O-ethoxymethyl modified nucleotide or 2'-O-2,2,2-trifluoroethoxymethyl modified nucleotide; Optionally, the 2'-O-(CH2) n -O-R1 modified nucleotides are selected from 2'-O-methoxyethyl modified nucleotides; Optionally, each nucleotide in the double-stranded oligonucleotide is independently selected from the following modified nucleotides: 2'-fluoro-modified nucleotides, 2'-deoxy-modified nucleotides, 2'-O-methyl-modified nucleotides or 2'-O-methoxyethyl-modified nucleotides; Optionally, each nucleotide in the double-stranded oligonucleotide is independently selected from the following modified nucleotides: 2'-Fluoro modified nucleotides, 2'-O-methyl modified nucleotides or 2'-O-methoxyethyl modified nucleotides.

6. The double-stranded oligonucleotide according to claim 5, characterized in that The double-stranded oligonucleotide contains at least one 2'-O-methoxyethyl modified nucleotide; Optionally, the antisense strand contains at least one 2'-O-methoxyethyl modified nucleotide; the sense strand contains at most one 2'-O-methoxyethyl modified nucleotide; Optionally, in the direction from the 5' end to the 3' end, at least three nucleotides in positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, at most one nucleotide in positions 5, 12, 13, and 18 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides in the remaining positions are independently selected from 2'-O-methyl-modified nucleotides; at least four nucleotides in positions 2, 6, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, and any one nucleotide in positions 9 to 12 is selected from 2'-fluoro-modified nucleotides, at least one nucleotide in positions 8 and 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides in the remaining positions are independently selected from 2'-O-methyl-modified nucleotides; Optionally, in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; At least four of the nucleotides at positions 2, 6, 14 and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, and any one of the nucleotides at positions 9 to 12 is selected from 2'-fluoro-modified nucleotides, at least one of the nucleotides at positions 8 and 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; Optionally, in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; At least four of the nucleotides at positions 2, 6, 14 and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, and any one of the nucleotides at positions 9 to 12 is selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 or 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; Optionally, the modifications of the sense strand and the antisense strand in the double-stranded oligonucleotide are selected from one of the following (ds-1) to (ds-8): (ds-1) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; The nucleotides at positions 2, 6, 9, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; (ds-2) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; The nucleotides at positions 2, 6, 10, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl modified nucleotides; (ds-3) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; The nucleotides at positions 2, 6, 11, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl modified nucleotides; (ds-4) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 in the nucleotide sequence of the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; The nucleotides at positions 2, 6, 12, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 8 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; (ds-5) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; The nucleotides at positions 2, 6, 9, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl modified nucleotides; (ds-6) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; The nucleotides at positions 2, 6, 10, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl modified nucleotides; (ds-7) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 in the nucleotide sequence of the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; The nucleotides at positions 2, 6, 11, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; (ds-8) in the direction from the 5' end to the 3' end, the nucleotides at positions 7 to 10 of the nucleotide sequence in the sense strand are selected from 2'-fluoro-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; The nucleotides at positions 2, 6, 12, 14, and 16 of the nucleotide sequence in the antisense strand are selected from 2'-fluoro-modified nucleotides, the nucleotide at position 15 is selected from 2'-O-methoxyethyl-modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-O-methyl-modified nucleotides; Optionally, the sense strand comprises two consecutive phosphorothioate internucleotide bonds at the 5' terminus; The antisense strand comprises two phosphorothioate internucleotide linkages at the 5' terminus and two consecutive phosphorothioate internucleotide linkages at the 3' terminus.

7. The double-stranded oligonucleotide according to claim 5, characterized in that Each nucleotide in the double-stranded oligonucleotide is independently selected from modified nucleotides; Wherein, the sense strand is selected from any nucleotide sequence of SEQ ID No.887-907; and / or the antisense strand is selected from any nucleotide sequence of SEQ ID No.908-943.

8. The double-stranded oligonucleotide according to claim 7, characterized in that The double-stranded oligonucleotide comprises one or more of the following:

9. The double-stranded oligonucleotide according to claim 7, characterized in that The sense strand is selected from any one of nucleotide sequences of SEQ ID No. 887, 892, 897, 902 and 907; and / or The antisense strand is selected from any one of the nucleotide sequences of SEQ ID No.908, 912, 914, 920, 927-943.

10. The double-stranded oligonucleotide according to claim 9, wherein The double-stranded oligonucleotide comprises one or more of the following:

11. The double-stranded oligonucleotide according to claim 10, wherein The double-stranded oligonucleotide is selected from the following group:

12. A double-stranded oligonucleotide for inhibiting C3 gene expression, characterized in that: The double-stranded oligonucleotide comprises a sense strand and an antisense strand, and each nucleotide in the double-stranded oligonucleotide is independently selected from modified nucleotides; Wherein, positions 2 to 19 of the antisense strand contain at least 17 consecutive nucleotides in any one of SEQ ID Nos. 908 to 943, or a nucleotide sequence having 1, 2 or 3 nucleotide base differences with the consecutive nucleotides; The sense strand contains at least 17 nucleotides, and the sense strand and the antisense strand are complementary or substantially complementary to form a duplex region; the substantially complementary means that the mismatch between the sense strand and the antisense strand in the duplex region does not exceed 3 nucleotides.

13. The double-stranded oligonucleotide according to claim 12, characterized in that The antisense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 908 to 943 and has 0, 1, 2 or 3 nucleotide base differences; Optionally, the antisense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 908 to 943 and having 0 or 1 nucleotide base difference; Optionally, the antisense strand comprises any one of the nucleotide sequences of SEQ ID Nos. 908 to 943.

14. The double-stranded oligonucleotide according to claim 12, wherein The sense strand comprises at least 17 consecutive nucleotides in any one of SEQ ID Nos. 887 to 907, or a nucleotide sequence having 1, 2 or 3 nucleotide base differences with the consecutive nucleotides; Optionally, the sense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 887 to 907 and having 0, 1, 2 or 3 nucleotide base differences; Optionally, the sense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 887 to 907 and having 0 or 1 nucleotide base difference; Optionally, the sense strand comprises any one of the nucleotide sequences of SEQ ID Nos. 887 to 907.

15. The double-stranded oligonucleotide according to any one of claims 12 to 14, characterized in that The double-stranded oligonucleotide comprises one or more of the following: An antisense strand having a nucleotide sequence as shown in SEQ ID No.908 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.909 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.910 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.909 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.888 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.909 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.889 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.909 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.890 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.909 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.891 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.911 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.888 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.911 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.889 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.911 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.890 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.911 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.891 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.912 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.913 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.914 and having 0 or 1 nucleotide base difference therefrom; and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 or having 0 or 1 nucleotide difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.913 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.893 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.913 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.894 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.913 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.895 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.913 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.896 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.915 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.893 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.915 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.894 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.915 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.895 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.915 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.896 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.916 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.917 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.918 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.917 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.898 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.917 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.899 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 917 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 900 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 917 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 901 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.919 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.898 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.919 and having 0 or 1 nucleotide base difference therefrom; and a sense strand having a nucleotide sequence as shown in SEQ ID No.899 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 919 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 900 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.919 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.901 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 920 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 921 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 922 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 921 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 903 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 921 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 904 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.921 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.905 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.921 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.906 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 923 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 903 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 923 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 904 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 923 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 905 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 923 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 906 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.910 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.888 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.910 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.890 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.924 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.890 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.924 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.891 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.914 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.895 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.925 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.896 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 922 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 905 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 926 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 906 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.927 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 928 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 929 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 930 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.931 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.932 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.933 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 934 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 935 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 936 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.937 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.938 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 939 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 940 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 897 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.941 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.942 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 943 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 897 and having 0 or 1 nucleotide base difference therefrom; Optionally, the double-stranded oligonucleotide comprises one or more of the following: an antisense strand having a nucleotide sequence as shown in SEQ ID No.908, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887; an antisense strand having a nucleotide sequence as shown in SEQ ID No.909, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887; An antisense strand having a nucleotide sequence as shown in SEQ ID No.910, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887; An antisense strand having a nucleotide sequence as shown in SEQ ID No.909, and a sense strand having a nucleotide sequence as shown in SEQ ID No.888; an antisense strand having a nucleotide sequence as shown in SEQ ID No.909, and a sense strand having a nucleotide sequence as shown in SEQ ID No.889; an antisense strand having a nucleotide sequence as shown in SEQ ID No.909, and a sense strand having a nucleotide sequence as shown in SEQ ID No.890; an antisense strand having a nucleotide sequence as shown in SEQ ID No.909, and a sense strand having a nucleotide sequence as shown in SEQ ID No.891; an antisense strand having a nucleotide sequence as shown in SEQ ID No.911, and a sense strand having a nucleotide sequence as shown in SEQ ID No.888; an antisense strand having a nucleotide sequence as shown in SEQ ID No.911, and a sense strand having a nucleotide sequence as shown in SEQ ID No.889; an antisense strand having a nucleotide sequence as shown in SEQ ID No.911, and a sense strand having a nucleotide sequence as shown in SEQ ID No.890; An antisense strand having a nucleotide sequence as shown in SEQ ID No.911, and a sense strand having a nucleotide sequence as shown in SEQ ID No.891; An antisense strand having a nucleotide sequence as shown in SEQ ID No.912, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892; An antisense strand having a nucleotide sequence as shown in SEQ ID No.913, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892; An antisense strand having a nucleotide sequence as shown in SEQ ID No.914, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892; An antisense strand having a nucleotide sequence as shown in SEQ ID No.913, and a sense strand having a nucleotide sequence as shown in SEQ ID No.893; An antisense strand having a nucleotide sequence as shown in SEQ ID No.913, and a sense strand having a nucleotide sequence as shown in SEQ ID No.894; an antisense strand having a nucleotide sequence as shown in SEQ ID No.913, and a sense strand having a nucleotide sequence as shown in SEQ ID No.895; An antisense strand having a nucleotide sequence as shown in SEQ ID No.913, and a sense strand having a nucleotide sequence as shown in SEQ ID No.896; An antisense strand having a nucleotide sequence as shown in SEQ ID No.915, and a sense strand having a nucleotide sequence as shown in SEQ ID No.893; An antisense strand having a nucleotide sequence as shown in SEQ ID No.915, and a sense strand having a nucleotide sequence as shown in SEQ ID No.894; An antisense strand having a nucleotide sequence as shown in SEQ ID No.915, and a sense strand having a nucleotide sequence as shown in SEQ ID No.895; an antisense strand having a nucleotide sequence as shown in SEQ ID No.915, and a sense strand having a nucleotide sequence as shown in SEQ ID No.896; an antisense strand having a nucleotide sequence as shown in SEQ ID No.916, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897; An antisense strand having a nucleotide sequence as shown in SEQ ID No.917, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897; An antisense strand having a nucleotide sequence as shown in SEQ ID No.918, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897; An antisense strand having a nucleotide sequence as shown in SEQ ID No.917, and a sense strand having a nucleotide sequence as shown in SEQ ID No.898; An antisense strand having a nucleotide sequence as shown in SEQ ID No.917, and a sense strand having a nucleotide sequence as shown in SEQ ID No.899; An antisense strand having a nucleotide sequence as shown in SEQ ID No.917, and a sense strand having a nucleotide sequence as shown in SEQ ID No.900; An antisense strand having a nucleotide sequence as shown in SEQ ID No.917, and a sense strand having a nucleotide sequence as shown in SEQ ID No.901; An antisense strand having a nucleotide sequence as shown in SEQ ID No.919, and a sense strand having a nucleotide sequence as shown in SEQ ID No.898; An antisense strand having a nucleotide sequence as shown in SEQ ID No.919, and a sense strand having a nucleotide sequence as shown in SEQ ID No.899; An antisense strand having a nucleotide sequence as shown in SEQ ID No.919, and a sense strand having a nucleotide sequence as shown in SEQ ID No.900; An antisense strand having a nucleotide sequence as shown in SEQ ID No.919, and a sense strand having a nucleotide sequence as shown in SEQ ID No.901; An antisense strand having a nucleotide sequence as shown in SEQ ID No.920, and a sense strand having a nucleotide sequence as shown in SEQ ID No.902; An antisense strand having a nucleotide sequence as shown in SEQ ID No.921, and a sense strand having a nucleotide sequence as shown in SEQ ID No.902; An antisense strand having a nucleotide sequence as shown in SEQ ID No.922, and a sense strand having a nucleotide sequence as shown in SEQ ID No.902; An antisense strand having a nucleotide sequence as shown in SEQ ID No.921, and a sense strand having a nucleotide sequence as shown in SEQ ID No.903; An antisense strand having a nucleotide sequence as shown in SEQ ID No.921, and a sense strand having a nucleotide sequence as shown in SEQ ID No.904; An antisense strand having a nucleotide sequence as shown in SEQ ID No.921, and a sense strand having a nucleotide sequence as shown in SEQ ID No.905; An antisense strand having a nucleotide sequence as shown in SEQ ID No.921, and a sense strand having a nucleotide sequence as shown in SEQ ID No.906; An antisense strand having a nucleotide sequence as shown in SEQ ID No.923, and a sense strand having a nucleotide sequence as shown in SEQ ID No.903; An antisense strand having a nucleotide sequence as shown in SEQ ID No.923, and a sense strand having a nucleotide sequence as shown in SEQ ID No.904; An antisense strand having a nucleotide sequence as shown in SEQ ID No.923, and a sense strand having a nucleotide sequence as shown in SEQ ID No.905; An antisense strand having a nucleotide sequence as shown in SEQ ID No.923, and a sense strand having a nucleotide sequence as shown in SEQ ID No.906; an antisense strand having a nucleotide sequence as shown in SEQ ID No.910, and a sense strand having a nucleotide sequence as shown in SEQ ID No.888; An antisense strand having a nucleotide sequence as shown in SEQ ID No.910, and a sense strand having a nucleotide sequence as shown in SEQ ID No.890; an antisense strand having a nucleotide sequence as shown in SEQ ID No.924, and a sense strand having a nucleotide sequence as shown in SEQ ID No.890; An antisense strand having a nucleotide sequence as shown in SEQ ID No.924, and a sense strand having a nucleotide sequence as shown in SEQ ID No.891; An antisense strand having a nucleotide sequence as shown in SEQ ID No.914, and a sense strand having a nucleotide sequence as shown in SEQ ID No.895; An antisense strand having a nucleotide sequence as shown in SEQ ID No.925, and a sense strand having a nucleotide sequence as shown in SEQ ID No.896; An antisense strand having a nucleotide sequence as shown in SEQ ID No.922, and a sense strand having a nucleotide sequence as shown in SEQ ID No.905; An antisense strand having a nucleotide sequence as shown in SEQ ID No.926, and a sense strand having a nucleotide sequence as shown in SEQ ID No.906; An antisense strand having a nucleotide sequence as shown in SEQ ID No.927, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892; An antisense strand having a nucleotide sequence as shown in SEQ ID No.928, and a sense strand having a nucleotide sequence as shown in SEQ ID No.907; An antisense strand having a nucleotide sequence as shown in SEQ ID No.929, and a sense strand having a nucleotide sequence as shown in SEQ ID No.907; An antisense strand having a nucleotide sequence as shown in SEQ ID No.930, and a sense strand having a nucleotide sequence as shown in SEQ ID No.907; An antisense strand having a nucleotide sequence as shown in SEQ ID No.931, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887; an antisense strand having a nucleotide sequence as shown in SEQ ID No.932, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887; An antisense strand having a nucleotide sequence as shown in SEQ ID No.933, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887; An antisense strand having a nucleotide sequence as shown in SEQ ID No.934, and a sense strand having a nucleotide sequence as shown in SEQ ID No.902; An antisense strand having a nucleotide sequence as shown in SEQ ID No.935, and a sense strand having a nucleotide sequence as shown in SEQ ID No.902; An antisense strand having a nucleotide sequence as shown in SEQ ID No.936, and a sense strand having a nucleotide sequence as shown in SEQ ID No.902; An antisense strand having a nucleotide sequence as shown in SEQ ID No.937, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892; An antisense strand having a nucleotide sequence as shown in SEQ ID No.938, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892; An antisense strand having a nucleotide sequence as shown in SEQ ID No.939, and a sense strand having a nucleotide sequence as shown in SEQ ID No.907; An antisense strand having a nucleotide sequence as shown in SEQ ID No.940, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897; An antisense strand having a nucleotide sequence as shown in SEQ ID No.941, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897; An antisense strand having a nucleotide sequence as shown in SEQ ID No.942, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897; The antisense strand has the nucleotide sequence shown in SEQ ID No.943, and the sense strand has the nucleotide sequence shown in SEQ ID No.

897.

16. The double-stranded oligonucleotide according to any one of claims 12 to 14, characterized in that The antisense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 908, 912, 914, 920, 927 to 943 and having 0, 1, 2 or 3 nucleotide base differences; Optionally, the antisense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 908, 912, 914, 920, 927 to 943 and having 0 or 1 nucleotide base difference; Optionally, the antisense strand comprises any one of the nucleotide sequences of SEQ ID Nos. 908, 912, 914, 920, 927 to 943.

17. The double-stranded oligonucleotide according to any one of claims 12 to 14, wherein the sense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 887, 892, 897, 902 and 907 and has 0, 1, 2 or 3 nucleotide base differences; Optionally, the sense strand comprises a nucleotide sequence of any one of SEQ ID Nos. 887, 892, 897, 902 and 907 and having 0 or 1 nucleotide base difference; Optionally, the sense strand comprises any one of the nucleotide sequences of SEQ ID Nos. 887, 892, 897, 902 and 907.

18. The double-stranded oligonucleotide according to any one of claims 16 to 17, characterized in that The double-stranded oligonucleotide comprises one or more of the following: An antisense strand having a nucleotide sequence as shown in SEQ ID No.908 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.931 and having 0 or 1 nucleotide base difference therefrom; and a sense strand having a nucleotide sequence as shown in SEQ ID No.887 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.932 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.933 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.912 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.914 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.927 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 and having 0 or 1 nucleotide base difference therefrom An antisense strand having a nucleotide sequence as shown in SEQ ID No.937 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 and having 0 or 1 nucleotide base difference therefrom An antisense strand having a nucleotide sequence as shown in SEQ ID No.938 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 and having 0 or 1 nucleotide base difference therefrom An antisense strand having a nucleotide sequence as shown in SEQ ID No.940 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897 and having 0 or 1 nucleotide base difference therefrom An antisense strand having a nucleotide sequence as shown in SEQ ID No.941 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897 and having 0 or 1 nucleotide base difference therefrom An antisense strand having a nucleotide sequence as shown in SEQ ID No.942 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 943 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 897 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.920 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.902 and having 0 or 1 nucleotide base difference therefrom An antisense strand having a nucleotide sequence as shown in SEQ ID No.934 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.902 and having 0 or 1 nucleotide base difference therefrom An antisense strand having a nucleotide sequence as shown in SEQ ID No. 935 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 936 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 902 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 928 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No. 929 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom; An antisense strand having a nucleotide sequence as shown in SEQ ID No.930 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.907 and having 0 or 1 nucleotide base difference therefrom An antisense strand having a nucleotide sequence as shown in SEQ ID No. 939 and having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No. 907 and having 0 or 1 nucleotide base difference therefrom; Optionally, the double-stranded oligonucleotide comprises one or more of the following: an antisense strand having a nucleotide sequence as shown in SEQ ID No.908, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887; An antisense strand having a nucleotide sequence as shown in SEQ ID No.931, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887; an antisense strand having a nucleotide sequence as shown in SEQ ID No.932, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887; An antisense strand having a nucleotide sequence as shown in SEQ ID No.933, and a sense strand having a nucleotide sequence as shown in SEQ ID No.887; An antisense strand having a nucleotide sequence as shown in SEQ ID No.912, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892; An antisense strand having a nucleotide sequence as shown in SEQ ID No.914, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892; An antisense strand having a nucleotide sequence as shown in SEQ ID No.927, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 An antisense strand having a nucleotide sequence as shown in SEQ ID No.937, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 An antisense strand having a nucleotide sequence as shown in SEQ ID No.938, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 An antisense strand having a nucleotide sequence as shown in SEQ ID No.940, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897 An antisense strand having a nucleotide sequence as shown in SEQ ID No.941, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897 An antisense strand having a nucleotide sequence as shown in SEQ ID No.942, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897; An antisense strand having a nucleotide sequence as shown in SEQ ID No.943, and a sense strand having a nucleotide sequence as shown in SEQ ID No.897; An antisense strand having a nucleotide sequence as shown in SEQ ID No.920, and a sense strand having a nucleotide sequence as shown in SEQ ID No.902 An antisense strand having a nucleotide sequence as shown in SEQ ID No.934, and a sense strand having a nucleotide sequence as shown in SEQ ID No.902 An antisense strand having a nucleotide sequence as shown in SEQ ID No.935, and a sense strand having a nucleotide sequence as shown in SEQ ID No.902; An antisense strand having a nucleotide sequence as shown in SEQ ID No.936, and a sense strand having a nucleotide sequence as shown in SEQ ID No.902; An antisense strand having a nucleotide sequence as shown in SEQ ID No.928, and a sense strand having a nucleotide sequence as shown in SEQ ID No.907; An antisense strand having a nucleotide sequence as shown in SEQ ID No.929, and a sense strand having a nucleotide sequence as shown in SEQ ID No.907; An antisense strand having a nucleotide sequence as shown in SEQ ID No.930, and a sense strand having a nucleotide sequence as shown in SEQ ID No.907 The antisense strand has the nucleotide sequence shown in SEQ ID No.939, and the sense strand has the nucleotide sequence shown in SEQ ID No.

907.

19. The double-stranded oligonucleotide according to any one of claims 12 to 14, characterized in that The antisense strand comprises a nucleotide sequence shown in SEQ ID No. 914 and having 0, 1, 2 or 3 nucleotide base differences; Optionally, the antisense strand comprises the nucleotide sequence shown in SEQ ID No. 914 and having 0 or 1 nucleotide base difference; Optionally, the antisense strand comprises the nucleotide sequence shown in SEQ ID No.

914.

20. The double-stranded oligonucleotide according to claim 19, wherein The sense strand comprises the nucleotide sequence shown in SEQ ID No. 892 and having 0, 1, 2 or 3 nucleotide base differences; Optionally, the sense strand comprises the nucleotide sequence shown in SEQ ID No. 892 and having 0 or 1 nucleotide base difference; Optionally, the sense strand comprises the nucleotide sequence shown in SEQ ID No.

892.

21. The double-stranded oligonucleotide according to claim 20, characterized in that The double-stranded oligonucleotide comprises: An antisense strand having a nucleotide sequence as shown in SEQ ID No.914 or having 0 or 1 nucleotide base difference therefrom, and a sense strand having a nucleotide sequence as shown in SEQ ID No.892 or having 0 or 1 nucleotide base difference therefrom.

22. The double-stranded oligonucleotide according to claim 21, wherein The double-stranded oligonucleotide comprises: The antisense strand has the nucleotide sequence shown in SEQ ID No.914, and the sense strand has the nucleotide sequence shown in SEQ ID No.

892.

23. The double-stranded oligonucleotide according to any one of claims 1 to 22, characterized in that The double-stranded oligonucleotide is selected from siRNA.

24. A conjugate, characterized in that The conjugate comprises a double-stranded oligonucleotide as claimed in any one of claims 1 to 23 and one or more gene delivery vectors; Optionally, the gene delivery vector is selected from a ligand that binds to a cell surface receptor; Optionally, the ligand is selected from asialoglycoprotein receptor ligands (ASGPR ligands); Optionally, the ASGPR ligand comprises galactose or a derivative thereof; The galactose or its derivatives include but are not limited to galactose, galactosamine, N-formylgalactosamine, N-acetylgalactosamine, N-propionylgalactosamine, N-n-butyrylgalactosamine or N-isobutyrylgalactosamine; Optionally, the number of the ASGPR ligand is selected from one, and one of the ASGPR ligands is conjugated to the 3' end of the sense strand of the double-stranded oligonucleotide.

25. The conjugate according to claim 24, characterized in that The conjugate comprises a structure as shown in formula (101): Wherein, Nu represents the double-stranded oligonucleotide; Each of the R ligand independently selected from ASGPR ligands; The m is selected from 1, 2, 3 or 4; Optionally, m is selected from 1; Optionally, m is selected from 1; one of said R ligand Conjugated to the 3' end of the sense strand.

26. The conjugate according to claim 25, characterized in that Each of the R ligand Independently selected from the structure represented by formula (201) or a pharmaceutically acceptable salt or a stereoisomer thereof: Wherein, * represents the conjugation site of the ligand on the sense strand or the antisense strand; the j is selected from 1, 2, 3 or 4; Each of said Z is independently selected from hydroxyl or thiol; Each of said p is independently selected from 1, 2 or 3; Each of said q is independently selected from 1, 2 or 3; Each R2 is independently selected from H, optionally substituted C 1-6 Alkyl or optionally substituted C 1-6 Alkoxy; Each of said L is independently selected from optionally substituted C 2-20 Alkylene or R La and R Lb are independently selected from optionally substituted C 1-10 Alkylene, k is selected from 1, 2, 3, 4 or 5; Each Y is independently selected from O, S or NH; Optionally, m is selected from 1, 2 or 3; Optionally, said m is selected from 3; Optionally, Z is selected from hydroxyl; Optionally, said p is selected from 1; Optionally, q is selected from 1; Optionally, said R2 is selected from H; Optionally, each of said L is independently selected from optionally substituted C 2-10 Alkylene or Among them, R La and R Lb are independently selected from optionally substituted C 1-10 Alkylene, k is selected from 1, 2 or 3; Optionally, k is selected from 1; Optionally, each L is independently selected from Optionally, Y is selected from O.

27. The conjugate according to claim 26, characterized in that Each R ligand Independently selected from the structure shown in formula (202) or a pharmaceutically acceptable salt or a stereoisomer thereof:

28. The conjugate according to any one of claims 25 to 27, characterized in that Each R ligand Independently selected from the structure shown in formula (CR01008×3) or a pharmaceutically acceptable salt or a stereoisomer thereof: Optionally, each Rligand is independently selected from the structure shown in formula (CR01013×3) or a pharmaceutically acceptable salt or a stereoisomer thereof:

29. The conjugate according to claim 28, characterized in that The conjugate includes one or more of the following groups: Optionally, the conjugate comprises one or more of the following: Optionally, the conjugate comprises the following group:

30. Use of the double-stranded oligonucleotide according to any one of claims 1 to 23 or the conjugate according to any one of claims 24 to 29 in the preparation of a medicament for alleviating, preventing and / or treating a disease or condition mediated by the C3 gene; Optionally, the C3 gene-mediated disease or condition includes, but is not limited to, IgA nephropathy, atypical hemolytic uremic syndrome (aHUS), including paroxysmal nocturnal hemoglobinuria (PNH), C3 glomerulopathy, wolf nephritis and / or membranous nephritis.

31. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the double-stranded nucleotide according to any one of claims 1 to 23 and / or the conjugate according to any one of claims 24 to 29.

32. A kit, characterized in that The kit comprises the double-stranded oligonucleotide according to any one of claims 1 to 23 and / or the conjugate according to any one of claims 24 to 29.

33. A method for inhibiting C3 gene expression, characterized in that: It comprises administering the double-stranded oligonucleotide according to any one of claims 1 to 23, and / or the conjugate according to any one of claims 24 to 29, and / or the pharmaceutical combination according to claim 31 to a subject.

34. A method for alleviating, treating and / or preventing a C3-mediated disease or condition, characterized in that: It comprises administering to a subject a double-stranded oligonucleotide according to any one of claims 1 to 23, and / or a conjugate according to any one of claims 24 to 29, and / or a pharmaceutical combination according to claim 31; Optionally, the C3-mediated disease or disorder comprises a disease associated with the level of mRNA expressed by the C3 gene; Optionally, the disease or disorder comprises IgA nephropathy, atypical hemolytic uremic syndrome, paroxysmal nocturnal hemoglobinuria (PNH), C3 glomerulopathy, lupus nephritis, or membranous nephritis.

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