Sirna and composition thereof

Abstract

Provided is an siRNA that inhibits the expression of an MMP7 gene, which siRNA contains a sense strand and an antisense strand. The provided siRNA has good stability, excellent MMP7 gene inhibitory activity, and negligible cytotoxicity and immunostimulation, and can significantly reduce MMP7 protein levels at the animal level. The siRNA has important clinical values in the treatment of diseases or conditions benefiting from the reduction of MMP7 levels or the inhibition of MMP7 expression.

Description

siRNA and its composition

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to Chinese patent application No. 202310944130.X filed on July 28, 2023 and Chinese patent application No. 202410176581.8 filed on February 8, 2024, the entire contents of which are incorporated herein by reference in their entirety. Technical Field

[0003] The present invention provides an siRNA for inhibiting the expression of matrix metalloproteinase 7 (MMP7) and a pharmaceutical composition thereof. The siRNA and the pharmaceutical composition provided by the present invention can treat diseases related to matrix metalloproteinase 7. Technical Background

[0004] Matrix metalloproteinase 7 (MMP7) is the smallest member of the matrix metalloproteinase (MMP) family, which consists of 24 related secreted zinc-dependent endopeptidases with diverse substrates and functions, capable of degrading components of the extracellular matrix and cleaving and regulating the activity of non-extracellular matrix substrates. Despite their attractiveness as drug targets, the development of MMP inhibitors is challenging due to the structural similarity of the zinc-dependent catalytic domains among family members. MMP7 is constitutively expressed and secreted by epithelial cells throughout the body and plays a role in epithelial repair. Increased MMP7 expression is associated with pathogenic fibrosis in the lung and liver. MMP7 enzyme levels are implicated in pathogenic fibrosis through multiple potential mechanisms, including promotion of epithelial-mesenchymal transition, extracellular matrix degradation, aberrant matrix repair, and tissue remodeling. MMP7 activates epithelial cells by cleaving E-cadherin and proteolytically activates heparin-binding epidermal growth factor precursor to release active HB-EGF, promoting epithelial-mesenchymal transition in epithelial cells and proliferation of lung fibroblasts, respectively, ultimately leading to or promoting fibrosis.

[0005] Idiopathic pulmonary fibrosis (IPF) is a chronic, often fatal lung disease with a relatively unpredictable clinical course and rate of disease progression. Increased expression of MMP7 has been observed in peripheral blood, bronchoalveolar lavage fluid, and lung tissue in IPF patients. Serum MMP7 expression is a serum biomarker for IPF and correlates with the severity and progression of IPF. Currently, IPF patients have only two drug options, pirfenidone and nintedanib, and the median survival of patients receiving these drugs remains short (only approximately 3 years), indicating the need for more effective drug options.

[0006] Small interfering RNA (siRNA) is an emerging and highly promising candidate for research. It can inhibit or block the expression of a target gene of interest in a sequence-specific manner based on the RNA interference mechanism, thereby achieving the purpose of treating diseases. Specifically, siRNA is loaded into the RNA-induced silencing complex (RISC), and its guide strand, also known as the antisense strand, is complementary paired with the target nucleic acid in the mRNA of the target gene, causing the mRNA of the target gene to degrade, thereby inhibiting or blocking the expression of the target gene. Since siRNA recognizes the target RNA in a sequence-specific manner through base complementary pairing, thereby achieving the cutting of the target RNA, achieving the purpose of lowering the target RNA level and inhibiting the expression of the target gene, siRNA has a very broad application prospect.

[0007] However, the interspecies differences in MMP7 increase the difficulty of siRNA drug development. Furthermore, compared to traditional drugs, siRNA is less stable and susceptible to nuclease degradation when administered systemically. Furthermore, efforts are needed to further enhance activity while avoiding side effects such as off-target effects, immune stimulation, and cytotoxicity. Therefore, developing more candidate siRNAs that inhibit MMP7 gene expression that are stable in the blood, have good biological activity, and have low cytotoxicity has become an urgent issue. Furthermore, using these candidate siRNAs to inhibit MMP7 gene expression to develop drugs that can effectively prevent and / or treat MMP7-related diseases requires clinical research and is commercially viable.

[0008] Summary of the Invention

[0009] The present invention provides siRNA that can effectively inhibit the expression of MMP7 gene, and thus provides drugs and methods for preventing and / or treating MMP7-related diseases.

[0010] siRNA

[0011] In one aspect, the present invention provides a small interfering RNA (siRNA) for inhibiting MMP7 gene expression, the siRNA comprising a sense strand and an antisense strand, wherein the antisense strand comprises at least 17 consecutive nucleotides that differ from the nucleotide sequence shown in any one of SEQ ID NO: 1 to SEQ ID NO: 124 by no more than 4 (e.g., 0, 1, 2, 3 or 4) nucleotides, and the sense strand is at least partially complementary to the antisense strand.

[0012] The at least partially complementary means that the two sequences can be fully complementary, or generally have no more than 6, 5, 4, 3, 2 or 1 mismatched base pairings, while retaining the ability to hybridize under relevant conditions. Those skilled in the art can determine the conditions that are most suitable for testing the complementarity of the two sequences based on the final application of the hybridized nucleotides. Such conditions can, for example, be stringent conditions, such as 400mM NaCl, 40mM PIPES pH 6.4, 1mM EDTA, 50°C or 70°C for 12-16 hours, followed by washing. Other conditions, such as physiologically relevant conditions that may be encountered in vivo, can also be applied.

[0013] In some embodiments, the antisense strand comprises at least 17 consecutive nucleotides that differ by 0 or 1 nucleotide from the nucleotide sequence shown in any one of SEQ ID NO: 1 to SEQ ID NO: 124.

[0014] In some embodiments, the antisense strand comprises at least 17 consecutive nucleotides that differ from the nucleotide sequence set forth in any one of SEQ ID NO: 1 to SEQ ID NO: 124 by no more than 4 (e.g., 0, 1, 2, 3, or 4) nucleotides. In some embodiments, the antisense strand comprises at least 17 consecutive nucleotides that differ from the nucleotide sequence set forth in any one of SEQ ID NO: 1 to SEQ ID NO: 124 by no more than 3 (e.g., 0, 1, 2, or 3) nucleotides. In some embodiments, the antisense strand comprises at least 17 consecutive nucleotides that differ from the nucleotide sequence set forth in any one of SEQ ID NO: 1 to SEQ ID NO: 124 by no more than 2 (e.g., 0, 1, or 2) nucleotides. In some embodiments, the antisense strand comprises at least 17 consecutive nucleotides that differ from the nucleotide sequence set forth in any one of SEQ ID NO: 1 to SEQ ID NO: 124 by no more than 1 (e.g., 0 or 1) nucleotide. In some embodiments, the antisense strand comprises at least 17 consecutive nucleotides of the nucleotide sequence shown in any one of SEQ ID NO: 1 to SEQ ID NO: 124. In some embodiments, the antisense strand is the nucleotide sequence shown in any one of SEQ ID NO: 1 to SEQ ID NO: 124.

[0015] In some embodiments, the sense strand has no more than 6 nucleotide mismatches with the antisense strand. In some embodiments, the sense strand has no more than 5 nucleotide mismatches with the antisense strand. In some embodiments, the sense strand has no more than 4 nucleotide mismatches with the antisense strand. In some embodiments, the sense strand has no more than 3 nucleotide mismatches with the antisense strand. In some embodiments, the sense strand has no more than 2 nucleotide mismatches with the antisense strand. In some embodiments, the sense strand has no more than 1 nucleotide mismatch with the antisense strand. In some embodiments, the sense strand is fully complementary to the antisense strand.

[0016] In some embodiments, the sense strand has at least 15, 16, 17, 18, 19, 20, or 21 nucleotides of complementarity to the antisense strand.

[0017] In some embodiments, the sense strand and the antisense strand are at least 85% (eg, at least 90%, at least 95%, at least 99%) complementary or fully complementary over at least 17 consecutive nucleotides.

[0018] In some embodiments, the sense strand and the antisense strand form a duplex region that is 15 to 30 nucleotide pairs in length, e.g., 15 to 25 nucleotide pairs, 15 to 23 nucleotide pairs, 15 to 21 nucleotide pairs, e.g., a duplex region of 15, 16, 17, 18, 19, 20, or 21 nucleotide pairs.

[0019] In some embodiments, the antisense strand is 17-30 (e.g., 17-29, 17-28, 17-27, 19-30, 19-29, 19-28, 19-27, 19-25, 19-23, 21-25, 21-23) nucleotides in length and the sense strand is 17-30 (e.g., 17-29, 17-28, 17-27, 17-26, 17-25, 17-21, 19-21) nucleotides in length.

[0020] In some embodiments, the antisense strand is 19 to 27 nucleotides in length; and the sense strand is 17 to 25 nucleotides in length.

[0021] In some embodiments, the antisense strand is 21 to 23 nucleotides in length; and the sense strand is 19 to 21 nucleotides in length.

[0022] In some embodiments, the antisense strand is 23 nucleotides in length and the sense strand is 21 nucleotides in length.

[0023] In some embodiments, the siRNA comprises blunt ends and / or overhangs.

[0024] In some embodiments, the siRNA comprises one or more single-stranded nucleotide overhangs. For example, an overhang of 1, 2, 3, or 4 nucleotides. In some embodiments, the overhang can be on the sense strand, the antisense strand, or any combination thereof. In some embodiments, the overhang is present on the 5' end, the 3' end, or both ends of the siRNA antisense strand or the sense strand.

[0025] In some embodiments, the 3' end of the antisense strand of the siRNA comprises a 2-nucleotide overhang. In some embodiments, the 3' end of the sense strand of the siRNA is blunt-ended.

[0026] In some embodiments, the sense strand of the siRNA comprises at least 17 consecutive nucleotides that are not more than 4 (e.g., 0, 1, 2, 3, or 4) nucleotides from the nucleotide sequence shown in any one of SEQ ID NO: 125 to SEQ ID NO: 248. The at least 17 consecutive nucleotides comprised by the sense strand of the siRNA and the antisense strand form a duplex region. In some embodiments, the sense strand comprises at least 17 consecutive nucleotides that are not more than 3 (e.g., 0, 1, 2, or 3) nucleotides from the nucleotide sequence shown in any one of SEQ ID NO: 125 to SEQ ID NO: 248. In some embodiments, the sense strand comprises at least 17 consecutive nucleotides that are not more than 2 (e.g., 0, 1, or 2) nucleotides from the nucleotide sequence shown in any one of SEQ ID NO: 125 to SEQ ID NO: 248. In some embodiments, the sense strand comprises at least 17 consecutive nucleotides that differ from the nucleotide sequence shown in any one of SEQ ID NO: 125 to SEQ ID NO: 248 by no more than 1 (e.g., 0 or 1) nucleotide. In some embodiments, the sense strand has a region of at least 85% (e.g., at least 90%, at least 95%, at least 99%) complementarity or complete complementarity to the antisense strand within the 17 consecutive nucleotides. In some embodiments, the sense strand comprises at least 17 consecutive nucleotides of the nucleotide sequence shown in any one of SEQ ID NO: 125 to SEQ ID NO: 248. In some embodiments, the sense strand is the nucleotide sequence shown in any one of SEQ ID NO: 125 to SEQ ID NO: 248.

[0027] In some embodiments, the antisense strand of the siRNA comprises the antisense strand sequence of any one of duplexes 1 to duplex 124 provided in Table 1, or a portion thereof (e.g., at least 17 consecutive nucleotides thereof), and the sense strand of the siRNA comprises the sense strand sequence of the duplex, or a portion thereof (e.g., at least 17 consecutive nucleotides thereof).

[0028] In some embodiments, the sequences of the sense strand and antisense strand of the siRNA are selected from the sense strand and antisense strand sequences of any duplex 1 to duplex 124 described in Table 1.

[0029] In some embodiments, the antisense strand of the siRNA comprises the antisense strand sequence of duplex 44, duplex 9, duplex 33, duplex 12, duplex 61, duplex 8, duplex 11, duplex 38, or duplex 73 provided in Table 1, or a portion thereof (e.g., at least 17 consecutive nucleotides thereof). Preferably, the sense strand of the siRNA comprises the sense strand sequence of the duplex, or a portion thereof (e.g., at least 17 consecutive nucleotides thereof).

[0030] In some embodiments, the antisense strand of the siRNA comprises the antisense strand sequence of duplex 27, duplex 85, duplex 86, duplex 20, duplex 65, duplex 2, duplex 5, duplex 14, duplex 58, duplex 4, duplex 52, duplex 95, duplex 103, duplex 3, duplex 67, duplex 54, duplex 111, duplex 46, duplex 108, or duplex 119 provided in Table 1, or a portion thereof (e.g., at least 17 consecutive nucleotides thereof). Preferably, the sense strand of the siRNA comprises the sense strand sequence of the duplex, or a portion thereof (e.g., at least 17 consecutive nucleotides thereof).

[0031] In some embodiments, the antisense strand of the siRNA comprises the antisense strand sequence of duplex 24, duplex 34, duplex 50, duplex 53, duplex 59, duplex 72, duplex 75, duplex 87, duplex 42, duplex 94, duplex 1, duplex 35, duplex 79, duplex 82, duplex 25, duplex 39, duplex 71, duplex 112, duplex 124, duplex 100, duplex 109, duplex 113, duplex 30, duplex 105, duplex 115, duplex 7, duplex 51, duplex 15, duplex 97, or duplex 121 provided in Table 1, or a portion thereof (e.g., at least 17 contiguous nucleotides thereof). Preferably, the sense strand of the siRNA comprises the sense strand sequence of the duplex or a portion thereof (eg, at least 17 consecutive nucleotides thereof).

[0032] In some embodiments, the antisense strand of the siRNA comprises the antisense strand sequence of duplex 69, duplex 81, duplex 88, duplex 96, duplex 22, duplex 32, duplex 47, duplex 40, duplex 21, duplex 29, duplex 31, duplex 45, duplex 63, duplex 18, duplex 70, duplex 114, duplex 123, duplex 41, duplex 90, duplex 122, duplex 84, or duplex 10 provided in Table 1, or a portion thereof (e.g., at least 17 consecutive nucleotides thereof). Preferably, the sense strand of the siRNA comprises the sense strand sequence of the duplex, or a portion thereof (e.g., at least 17 consecutive nucleotides thereof).

[0033] In some embodiments, the antisense strand of the siRNA comprises the antisense strand sequence of duplex 6, duplex 60, duplex 80, duplex 118, duplex 16, duplex 55, duplex 116, duplex 19, duplex 120, duplex 13, duplex 74, duplex 23, duplex 56, or duplex 76 provided in Table 1, or a portion thereof (e.g., at least 17 consecutive nucleotides thereof). Preferably, the sense strand of the siRNA comprises the sense strand sequence of the duplex, or a portion thereof (e.g., at least 17 consecutive nucleotides thereof).

[0034] In some embodiments, the antisense strand of the siRNA comprises the antisense strand sequence of duplex 2, duplex 5, duplex 8, duplex 9, duplex 11, duplex 12, duplex 33, duplex 35, duplex 38, duplex 46, duplex 50, duplex 67, or duplex 73 provided in Table 1, or a portion thereof (e.g., at least 17 consecutive nucleotides thereof). Preferably, the sense strand of the siRNA comprises the sense strand sequence of the duplex, or a portion thereof (e.g., at least 17 consecutive nucleotides thereof).

[0035] In some embodiments, the antisense strand of the siRNA comprises the antisense strand sequence of duplex 2, duplex 9, duplex 11, duplex 12, duplex 38, duplex 46, duplex 67, or duplex 73 provided in Table 1, or a portion thereof (e.g., at least 17 consecutive nucleotides thereof). Preferably, the sense strand of the siRNA comprises the sense strand sequence of the duplex, or a portion thereof (e.g., at least 17 consecutive nucleotides thereof).

[0036] Modification

[0037] The siRNA of the present invention can be modified in nucleoside base structure or in ribose-phosphate backbone structure, reduce the effect of missing the target, and / or increase the biological stability of the molecule, or increase the physical stability of the duplex formed between antisense and sense nucleic acid. Therefore, the siRNA sequence comprising any modification is also encompassed within the scope of the present invention. The siRNA molecule comprising ribonucleoside analogs or derivatives must maintain the ability to form duplexes and allow or mediate the specific degradation of the target RNA via the RISC approach.

[0038] In some embodiments of the invention, the siRNA contains at least one modified nucleotide. The modification need not be identical for each of the plurality of modified ribonucleosides in the siRNA.

[0039] In some embodiments of the present invention, all nucleotides in the sense strand and / or antisense strand of the siRNA are modified nucleotides or nucleotide analogs.

[0040] In some embodiments of the invention, the siRNA comprises 2'-modified nucleotides.

[0041] In some embodiments of the present invention, the modified nucleotide is selected from 2'-methoxy nucleotides, 2'-fluoro nucleotides, 2'-deoxy nucleotides, 2',3'-split nucleotide analogs, 2'-fluoroarabino nucleotides, 2'-methoxyethyl nucleotides, 2'-amino modified nucleotides, 2'-alkyl modified nucleotides, 3'-methoxy nucleotides, 2'-allyl modified nucleotides, nucleotides containing thiophosphate groups, nucleotides containing methylphosphonate groups, nucleotides containing 5'-phosphate esters, nucleotides containing 5'-phosphate mimetics, diol-modified nucleotides, abasic nucleotides, morpholino nucleotides, locked nucleotides (LNA), unlocked nucleotides (UNA) or glycerol nucleotides (GNA), but the present invention is not limited thereto.

[0042] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length and comprises:

[0043] 1)AS(5'-3'):

[0044] mN*fN*mNmNmNfNmNfNfNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 380);

[0045] 2)AS(5'-3'):

[0046] (SEQ ID NO:381);

[0047] 3) AS(5'-3'):

[0048] mN*fN*mNfNmNmNmNmNmNmNmNmNmNfNmNfNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:382);

[0049] 4) AS(5'-3'):

[0050] mN*fN*mNmNmNfNmNfNfNfNfNmNmNmNfNmNfNmNmNmNmNmN*mN*mN(SEQ ID NO:383);

[0051] 5) AS(5'-3'):

[0052] mN*fN*mNfNmNfNmNfNmNmNmNmNmNmNfNmNfNmNfNmNfNmN*mN*mN(SEQ ID NO:384);

[0053] 6) AS(5'-3'):

[0054] mN*fN*mNmNmNfNmNmNmNmNmNmNmNmNfNmNfNfNmNmNmNmN*mN*mN(SEQ ID NO:385);

[0055] 7) AS(5'-3'):

[0056] mN*fN*mNmNmNmNmNfNfNmNmNmNmNmNfNfNmNmNmNmNmNmN*mN*mN(SEQ ID NO:386);

[0057] 8) AS(5'-3'):

[0058] mN*fN*mNfNmNfNmNfNmNfNmNmNmNfNmNfNmNfNmNfNmN*mN*mN(SEQ ID NO:387);

[0059] 9) AS(5'-3'):

[0060] (E)-VPmN*fN*mNmNmNmNmNfNfNmNmNmNmNmNfNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:388);

[0061] 10)AS(5'-3'):

[0062] (E)-VPmN*fN*mNmNmNfNmNfNfNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 389);

[0063] 11)AS(5'-3'):

[0064] (E)-VPmN*fN*mNmNmNmNmNmNfNmNmNfNmNfNmNfNmNmNmNmNmN*mN*m N (SEQ ID NO: 390); or,

[0065] 12)AS(5'-3'):

[0066] (E)-VPmN*fN*mNmNmNfNmNfNfNfNmNmNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 391);

[0067] Wherein, mN is a methoxy-modified nucleotide, fN is a fluorinated-modified nucleotide, (E)-VP indicates that the nucleotide adjacent to its right is a (E)-vinylphosphonate-modified nucleotide, and * indicates a phosphorothioate linkage.

[0068] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length and comprises:

[0069] 1) The 2nd, 6th, 8th, 14th, and 16th positions on the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; or

[0070] 2) the 5' end of the antisense strand has 2'-fluoro nucleotides at positions 2, 6, 14, 16, and 18, and the remaining positions have 2'-methoxy nucleotides; or

[0071] 3) the 2nd, 6th, 14th, 16th, and 20th positions on the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; or

[0072] 4) the 5' end of the antisense strand has 2'-fluoro nucleotides at positions 2, 6, 8, 9, 12, 14, and 16, and the remaining positions have 2'-methoxy nucleotides; or

[0073] 5) 2'-fluoro nucleotides at positions 2, 9, 12, 14, and 16 of the 5' end of the antisense strand, and 2'-methoxy nucleotides at the remaining positions; or

[0074] 6) the 2nd, 4th, 12th, 14th, and 16th positions on the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; or

[0075] 7) the 5' end of the antisense strand has 2'-fluoro nucleotides at positions 2, 6, 8, 9, 10, 14, and 16, and the remaining positions have 2'-methoxy nucleotides; or

[0076] 8) the 5' end of the antisense strand has 2'-fluoro nucleotides at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20, and the remaining positions have 2'-methoxy nucleotides; or

[0077] 9) the 5' end of the antisense strand has 2'-fluoro nucleotides at positions 2, 8, 9, 14, and 16, and the remaining positions have 2'-methoxy nucleotides; or

[0078] 10) The 2nd, 4th, 6th, 8th, 14th, 16th, 18th, and 20th positions on the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0079] In some embodiments of the present invention, the sense strand of the siRNA is 21 nucleotides in length and comprises:

[0080] 1) SS (5'-3'):

[0081] mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmNmNmNmN (SEQ ID NO: 392);

[0082] 2) SS (5'-3'):

[0083] mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmN (SEQ ID NO: 393);

[0084] 3) SS (5'-3'):

[0085] mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNmNmNmNmNmNmN (SEQ ID NO: 394);

[0086] 4) SS (5'-3'):

[0087] mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmNmNmN (SEQ ID NO: 395);

[0088] 5)SS(5'-3'):

[0089] mN*mN*mNmNmNmNfNmNfNfNfNmNfNmNmNmNmNmNmNmNmNmN(SEQ ID NO:396);

[0090] 6)SS(5'-3'):

[0091] mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNmNmNmNmNmN(SEQ ID NO:397);

[0092] 7)SS(5'-3'):

[0093] mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNfNfNmNmNmNmN(SEQ ID NO:398);

[0094] 8)SS(5'-3'):

[0095] mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNfNmNmNmN*mN(SEQ ID NO:399);

[0096] 9)SS(5'-3'):

[0097] mN*mN*mNmNmNmNfNmNdNfNfNmNmNmNmNfNmNmNmN*mN(SEQ ID NO:400);

[0098] 10)SS(5'-3'):

[0099] mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNfNfNmNmNmN*mN(SEQ ID NO:401);

[0100] 11)SS(5'-3'):

[0101] mN*mN*mNmNmNmNmNmNfNfNfNmNmNmNmNfNfNmNmNmN*mN(SEQ ID NO:402);

[0102] 12)SS(5'-3'):

[0103] mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNfNfNmNmNmN*mN(SEQ ID NO:403);

[0104] 13)SS(5'-3'):

[0105] mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNfNfNmNmN*mN*mN(SEQ ID NO:404);

[0106] 14)SS(5'-3'):

[0107] mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNfNfNmNmN*mN(SEQ ID NO:405);

[0108] 15)SS(5'-3'):

[0109] mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNmNfNmNmNmN*mN(SEQ ID NO:406);

[0110] 16)SS(5'-3'):

[0111] mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmNmNmN*mN(SEQ ID NO:407);

[0112] 17)SS(5'-3'):

[0113] mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:408);

[0114] 18)SS(5'-3'):

[0115] mN*mN*mNmNmNmNmNmNmNmNmNfNmNfNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:409);

[0116] 19)SS(5'-3'):

[0117] mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:410);

[0118] 20)SS(5'-3'):

[0119] mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:411);

[0120] 21)SS(5'-3'):

[0121] mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmNmNmNmN*(iab) (SEQ ID NO: 412);

[0122] 22)SS(5'-3'):

[0123] mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmN*(iab) (SEQ ID NO: 413);

[0124] 23)SS(5'-3'):

[0125] mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmNmNmN*(iab) (SEQ ID NO: 414);

[0126] 24)SS(5'-3'):

[0127] mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmNmNmNmN*(iab) (SEQ ID NO: 415); or,

[0128] 25)SS(5'-3'):

[0129] mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNmNfNmNmNmNmN*(iab) (SEQ ID NO: 416);

[0130] Wherein, mN is a methoxy-modified nucleotide, fN is a fluorinated-modified nucleotide, dN is a deoxynucleotide, * is a phosphorothioate linkage, and (iab) is an inverted abasic residue.

[0131] In some embodiments of the present invention, the sense strand of the siRNA is 21 nucleotides in length and comprises:

[0132] 1) The 7th, 9th, and 11th positions on the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; or

[0133] 2) the 7th, 9th, and 10th positions of the 5' end of the sense strand are 2'-fluoro nucleotides, the 11th position is a deoxy nucleotide, and the remaining positions are 2'-methoxy nucleotides; or

[0134] 3) the 7th, 9th, 10th, 11th, and 13th positions on the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; or

[0135] 4) positions 7, 9, 10, 11, and 14 of the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; or

[0136] 5) the 7th, 9th, 10th, 11th, and 15th positions on the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; or

[0137] 6) positions 7, 9, 10, 11, and 17 of the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; or

[0138] 7) positions 7, 9, 10, 11, and 19 of the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; or

[0139] 8) the 9th, 11th, and 13th positions on the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; or

[0140] 9) the 7th, 9th, 10th, 11th, and 16th positions on the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; or

[0141] 10) The 5' end of the sense strand has 2'-fluoro nucleotides at positions 7, 9, 11, and 17, a deoxynucleotide at position 10, and 2'-methoxy nucleotides at the remaining positions; or

[0142] 11) The 7th, 9th, and 11th positions of the 5' end of the sense strand are 2'-fluoro nucleotides, the 10th position is a deoxy nucleotide, and the remaining positions are 2'-methoxy nucleotides; or

[0143] 12) The 5' end of the sense strand has 2'-fluoro nucleotides at positions 7, 10, 11, and 17, a deoxynucleotide at position 9, and 2'-methoxy nucleotides at the remaining positions; or

[0144] 13) positions 7, 9, 10, and 17 of the 5' end of the sense strand are 2'-fluoro nucleotides, position 11 is a deoxy nucleotide, and the remaining positions are 2'-methoxy nucleotides; or

[0145] 14) positions 9, 10, 11, and 17 of the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; or

[0146] 15) positions 7, 9, 10, 11, 16, and 17 of the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; or

[0147] 16) Positions 7, 9, 10, 11, 15, and 17 of the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0148] In some embodiments, the siRNA comprises a modification pattern selected from the group consisting of: SEQ ID NO: 380 and SEQ ID NO: 392; SEQ ID NO: 380 and SEQ ID NO: 393; SEQ ID NO: 380 and SEQ ID NO: 394; SEQ ID NO: 380 and SEQ ID NO: 395; SEQ ID NO: 380 and SEQ ID NO: 396; SEQ ID NO: 380 and SEQ ID NO: 397; SEQ ID NO: 380 and SEQ ID NO: 398; SEQ ID NO: 380 and SEQ ID NO: 399; SEQ ID NO: 380 and SEQ ID NO: 400; SEQ ID NO: 380 and SEQ ID NO: 401; SEQ ID NO: 380 and SEQ ID NO: 402; SEQ ID NO: 380 and SEQ ID NO: 403; SEQ ID NO: 380 and SEQ ID NO: 404; SEQ ID NO: 380 and SEQ ID NO: 405; SEQ ID NO: 380 and SEQ ID NO: 406; SEQ ID NO: 380 and SEQ ID NO: 407; SEQ ID NO: 380 and SEQ ID NO: 408; SEQ ID NO: 380 and SEQ ID NO: 409; SEQ ID NO: 380 and SEQ ID NO: 410; SEQ ID NO: 380 and SEQ ID NO: 411; SEQ ID NO: 380 and SEQ ID NO: 412; SEQ ID NO: 380 and SEQ ID NO: 413; SEQ ID NO: 380 and SEQ ID NO: 414; SEQ ID NO: 380 NO:380 and SEQ ID NO:405; SEQ ID NO:380 and SEQ ID NO:406; SEQ ID NO:380 and SEQ ID NO:407; SEQ ID NO:380 and SEQ ID NO:408; SEQ ID NO:380 and SEQ ID NO:409; SEQ ID NO:380 and SEQ ID NO:410; SEQ ID NO:380 and SEQ ID NO:411; SEQ ID NO:380 and SEQ ID NO:412; SEQ ID NO:380 and SEQ ID NO:413; SEQ ID NO:380 and SEQ ID NO:414; SEQ ID NO:380 and SEQ ID NO:415; SEQ ID NO:380 and SEQ ID NO:416;

[0149] SEQ ID NO:381 and SEQ ID NO:392; SEQ ID NO:381 and SEQ ID NO:393; SEQ ID NO:381 and SEQ ID NO:394; SEQ ID NO:381 and SEQ ID NO:395; SEQ ID NO:381 and SEQ ID NO:396; SEQ ID NO:381 and SEQ ID NO:397; SEQ ID SEQ ID NO:381 and SEQ ID NO:398; SEQ ID NO:381 and SEQ ID NO:399; SEQ ID NO:381 and SEQ ID NO:400; SEQ ID NO:381 and SEQ ID NO:401; SEQ ID NO:381 and SEQ ID NO:402; SEQ ID NO:381 and SEQ ID NO:403; SEQ ID NO:381 and SEQ ID NO:404; SEQ ID NO:381 and SEQ ID NO:405; SEQ ID NO:381 and SEQ ID NO:406; SEQ ID NO:381 and SEQ ID NO:407; SEQ ID NO:381 and SEQ ID NO:408; SEQ ID NO:381 and SEQ ID NO:409; SEQ ID NO:381 and SEQ ID NO:410; SEQ ID NO:381 and SEQ ID NO:411; SEQ ID NO:381 and SEQ ID NO:412; SEQ ID NO:381 and SEQ ID NO:413; SEQ ID NO:381 and SEQ ID NO:414; SEQ ID NO:381 and SEQ ID NO:415; SEQ ID NO:381 and SEQ ID NO:416;

[0150] SEQ ID NO:382 and SEQ ID NO:392; SEQ ID NO:382 and SEQ ID NO:393; SEQ ID NO:382 and SEQ ID NO:394; SEQ ID NO:382 and SEQ ID NO:395; SEQ ID NO:382 and SEQ ID NO:396; SEQ ID NO:382 and SEQ ID NO:397; SEQ ID SEQ ID NO:382 and SEQ ID NO:398; SEQ ID NO:382 and SEQ ID NO:399; SEQ ID NO:382 and SEQ ID NO:400; SEQ ID NO:382 and SEQ ID NO:401; SEQ ID NO:382 and SEQ ID NO:402; SEQ ID NO:382 and SEQ ID NO:403; SEQ ID NO:382 and SEQ ID NO:382 and SEQ ID NO:402. NO:404; SEQ ID NO:382 and SEQ ID NO:405; SEQ ID NO:382 and SEQ ID NO:406; SEQ ID NO:382 and SEQ ID NO:407; SEQ ID NO:382 and SEQ ID NO:408; SEQ ID NO:382 and SEQ ID NO:409; SEQ ID NO:382 and SEQ ID NO:410; SEQ ID NO:382 and SEQ ID NO:411; SEQ ID NO:382 and SEQ ID NO:412; SEQ ID NO:382 and SEQ ID NO:413; SEQ ID NO:382 and SEQ ID NO:414; SEQ ID NO:382 and SEQ ID NO:415; SEQ ID NO:382 and SEQ ID NO:416;

[0151] SEQ ID NO:383 and SEQ ID NO:392; SEQ ID NO:383 and SEQ ID NO:393; SEQ ID NO:383 and SEQ ID NO:394; SEQ ID NO:383 and SEQ ID NO:395; SEQ ID NO:383 and SEQ ID NO:396; SEQ ID NO:383 and SEQ ID NO:397; SEQ ID SEQ ID NO:383 and SEQ ID NO:398; SEQ ID NO:383 and SEQ ID NO:399; SEQ ID NO:383 and SEQ ID NO:400; SEQ ID NO:383 and SEQ ID NO:401; SEQ ID NO:383 and SEQ ID NO:402; SEQ ID NO:383 and SEQ ID NO:403; SEQ ID NO:383 and SEQ ID NO:383 and SEQ ID NO:401. NO:404; SEQ ID NO:383 and SEQ ID NO:405; SEQ ID NO:383 and SEQ ID NO:406; SEQ ID NO:383 and SEQ ID NO:407; SEQ ID NO:383 and SEQ ID NO:408; SEQ ID NO:383 and SEQ ID NO:409; SEQ ID NO:383 and SEQ ID NO:410; SEQ ID NO:383 and SEQ ID NO:411; SEQ ID NO:383 and SEQ ID NO:412; SEQ ID NO:383 and SEQ ID NO:413; SEQ ID NO:383 and SEQ ID NO:414; SEQ ID NO:383 and SEQ ID NO:415; SEQ ID NO:383 and SEQ ID NO:416;

[0152] SEQ ID NO:384 and SEQ ID NO:392; SEQ ID NO:384 and SEQ ID NO:393; SEQ ID NO:384 and SEQ ID NO:394; SEQ ID NO:384 and SEQ ID NO:395; SEQ ID NO:384 and SEQ ID NO:396; SEQ ID NO:384 and SEQ ID NO:397; SEQ ID SEQ ID NO:384 and SEQ ID NO:398; SEQ ID NO:384 and SEQ ID NO:399; SEQ ID NO:384 and SEQ ID NO:400; SEQ ID NO:384 and SEQ ID NO:401; SEQ ID NO:384 and SEQ ID NO:402; SEQ ID NO:384 and SEQ ID NO:403; SEQ ID NO:384 and SEQ ID NO:384 and SEQ ID NO:401. NO:404; SEQ ID NO:384 and SEQ ID NO:405; SEQ ID NO:384 and SEQ ID NO:406; SEQ ID NO:384 and SEQ ID NO:407; SEQ ID NO:384 and SEQ ID NO:408; SEQ ID NO:384 and SEQ ID NO:409; SEQ ID NO:384 and SEQ ID NO:410; SEQ ID NO:384 and SEQ ID NO:411; SEQ ID NO:384 and SEQ ID NO:412; SEQ ID NO:384 and SEQ ID NO:413; SEQ ID NO:384 and SEQ ID NO:414; SEQ ID NO:384 and SEQ ID NO:415; SEQ ID NO:384 and SEQ ID NO:416;

[0153] SEQ ID NO:385 and SEQ ID NO:392; SEQ ID NO:385 and SEQ ID NO:393; SEQ ID NO:385 and SEQ ID NO:394; SEQ ID NO:385 and SEQ ID NO:395; SEQ ID NO:385 and SEQ ID NO:396; SEQ ID NO:385 and SEQ ID NO:397; SEQ ID SEQ ID NO:385 and SEQ ID NO:398; SEQ ID NO:385 and SEQ ID NO:399; SEQ ID NO:385 and SEQ ID NO:400; SEQ ID NO:385 and SEQ ID NO:401; SEQ ID NO:385 and SEQ ID NO:402; SEQ ID NO:385 and SEQ ID NO:403; SEQ ID NO:385 and SEQ ID NO:385 and SEQ ID NO:401. NO:404; SEQ ID NO:385 and SEQ ID NO:405; SEQ ID NO:385 and SEQ ID NO:406; SEQ ID NO:385 and SEQ ID NO:407; SEQ ID NO:385 and SEQ ID NO:408; SEQ ID NO:385 and SEQ ID NO:409; SEQ ID NO:385 and SEQ ID NO:410; SEQ ID NO:385 and SEQ ID NO:411; SEQ ID NO:385 and SEQ ID NO:412; SEQ ID NO:385 and SEQ ID NO:413; SEQ ID NO:385 and SEQ ID NO:414; SEQ ID NO:385 and SEQ ID NO:415; SEQ ID NO:385 and SEQ ID NO:416;

[0154] SEQ ID NO:386 and SEQ ID NO:392; SEQ ID NO:386 and SEQ ID NO:393; SEQ ID NO:386 and SEQ ID NO:394; SEQ ID NO:386 and SEQ ID NO:395; SEQ ID NO:386 and SEQ ID NO:396; SEQ ID NO:386 and SEQ ID NO:397; SEQ ID NO:386 and SEQ ID NO:397; SEQ ID NO:386 and SEQ ID NO:398; SEQ ID NO:386 and SEQ ID NO:399; SEQ ID NO:386 and SEQ ID NO:400; SEQ ID NO:386 and SEQ ID NO:401; SEQ ID NO:386 and SEQ ID NO:402; SEQ ID NO:386 and SEQ ID NO:403; SEQ ID NO:386 and SEQ ID NO:386 and SEQ ID NO:401. NO:404; SEQ ID NO:386 and SEQ ID NO:405; SEQ ID NO:386 and SEQ ID NO:406; SEQ ID NO:386 and SEQ ID NO:407; SEQ ID NO:386 and SEQ ID NO:408; SEQ ID NO:386 and SEQ ID NO:409; SEQ ID NO:386 and SEQ ID NO:410; SEQ ID NO:386 and SEQ ID NO:411; SEQ ID NO:386 and SEQ ID NO:412; SEQ ID NO:386 and SEQ ID NO:413; SEQ ID NO:386 and SEQ ID NO:414; SEQ ID NO:386 and SEQ ID NO:415; SEQ ID NO:386 and SEQ ID NO:416;

[0155] SEQ ID NO:387 and SEQ ID NO:392; SEQ ID NO:387 and SEQ ID NO:393; SEQ ID NO:387 and SEQ ID NO:394; SEQ ID NO:387 and SEQ ID NO:395; SEQ ID NO:387 and SEQ ID NO:396; SEQ ID NO:387 and SEQ ID NO:397; SEQ ID SEQ ID NO:387 and SEQ ID NO:398; SEQ ID NO:387 and SEQ ID NO:399; SEQ ID NO:387 and SEQ ID NO:400; SEQ ID NO:387 and SEQ ID NO:401; SEQ ID NO:387 and SEQ ID NO:402; SEQ ID NO:387 and SEQ ID NO:403; SEQ ID NO:387 and SEQ ID NO:404; SEQ ID NO:387 and SEQ ID NO:405; SEQ ID NO:387 and SEQ ID NO:406; SEQ ID NO:387 and SEQ ID NO:407; SEQ ID NO:387 and SEQ ID NO:408; SEQ ID NO:387 and SEQ ID NO:409; SEQ ID NO:387 and SEQ ID NO:410; SEQ ID NO:387 and SEQ ID NO:411; SEQ ID NO:387 and SEQ ID NO:412; SEQ ID NO:387 and SEQ ID NO:413; SEQ ID NO:387 and SEQ ID NO:414; SEQ ID NO:387 and SEQ ID NO:415; SEQ ID NO:387 and SEQ ID NO:416;

[0156] SEQ ID NO:388 and SEQ ID NO:392; SEQ ID NO:388 and SEQ ID NO:393; SEQ ID NO:388 and SEQ ID NO:394; SEQ ID NO:388 and SEQ ID NO:395; SEQ ID NO:388 and SEQ ID NO:396; SEQ ID NO:388 and SEQ ID NO:397; SEQ ID SEQ ID NO:388 and SEQ ID NO:398; SEQ ID NO:388 and SEQ ID NO:399; SEQ ID NO:388 and SEQ ID NO:400; SEQ ID NO:388 and SEQ ID NO:401; SEQ ID NO:388 and SEQ ID NO:402; SEQ ID NO:388 and SEQ ID NO:403; SEQ ID NO:388 and SEQ ID NO:388 and SEQ ID NO:401 NO:404; SEQ ID NO:388 and SEQ ID NO:405; SEQ ID NO:388 and SEQ ID NO:406; SEQ ID NO:388 and SEQ ID NO:407; SEQ ID NO:388 and SEQ ID NO:408; SEQ ID NO:388 and SEQ ID NO:409; SEQ ID NO:388 and SEQ ID NO:410; SEQ ID NO:388 and SEQ ID NO:411; SEQ ID NO:388 and SEQ ID NO:412; SEQ ID NO:388 and SEQ ID NO:413; SEQ ID NO:388 and SEQ ID NO:414; SEQ ID NO:388 and SEQ ID NO:415; SEQ ID NO:388 and SEQ ID NO:416;

[0157] SEQ ID NO:389 and SEQ ID NO:392; SEQ ID NO:389 and SEQ ID NO:393; SEQ ID NO:389 and SEQ ID NO:394; SEQ ID NO:389 and SEQ ID NO:395; SEQ ID NO:389 and SEQ ID NO:396; SEQ ID NO:389 and SEQ ID NO:397; SEQ ID NO:389 and SEQ ID NO:397; SEQ ID NO:389 and SEQ ID NO:398; SEQ ID NO:389 and SEQ ID NO:399; SEQ ID NO:389 and SEQ ID NO:400; SEQ ID NO:389 and SEQ ID NO:401; SEQ ID NO:389 and SEQ ID NO:402; SEQ ID NO:389 and SEQ ID NO:403; SEQ ID NO:389 and SEQ ID NO:389 and SEQ ID NO:401. NO:404; SEQ ID NO:389 and SEQ ID NO:405; SEQ ID NO:389 and SEQ ID NO:406; SEQ ID NO:389 and SEQ ID NO:407; SEQ ID NO:389 and SEQ ID NO:408; SEQ ID NO:389 and SEQ ID NO:409; SEQ ID NO:389 and SEQ ID NO:410; SEQ ID NO:389 and SEQ ID NO:411; SEQ ID NO:389 and SEQ ID NO:412; SEQ ID NO:389 and SEQ ID NO:413; SEQ ID NO:389 and SEQ ID NO:414; SEQ ID NO:389 and SEQ ID NO:415; SEQ ID NO:389 and SEQ ID NO:416;

[0158] SEQ ID NO:390 and SEQ ID NO:392; SEQ ID NO:390 and SEQ ID NO:393; SEQ ID NO:390 and SEQ ID NO:394; SEQ ID NO:390 and SEQ ID NO:395; SEQ ID NO:390 and SEQ ID NO:396; SEQ ID NO:390 and SEQ ID NO:397; SEQ ID SEQ ID NO:390 and SEQ ID NO:398; SEQ ID NO:390 and SEQ ID NO:399; SEQ ID NO:390 and SEQ ID NO:400; SEQ ID NO:390 and SEQ ID NO:401; SEQ ID NO:390 and SEQ ID NO:402; SEQ ID NO:390 and SEQ ID NO:403; SEQ ID NO:390 and SEQ ID NO:404; SEQ ID NO:390 and SEQ ID NO:405; SEQ ID NO:390 and SEQ ID NO:406; SEQ ID NO:390 and SEQ ID NO:407; SEQ ID NO:390 and SEQ ID NO:408; SEQ ID NO:390 and SEQ ID NO:409; SEQ ID NO:390 and SEQ ID NO:410; SEQ ID NO:390 and SEQ ID NO:411; SEQ ID NO:390 and SEQ ID NO:412; SEQ ID NO:390 and SEQ ID NO:413; SEQ ID NO:390 and SEQ ID NO:414; SEQ ID NO:390 and SEQ ID NO:415; SEQ ID NO:390 and SEQ ID NO:416;

[0159] SEQ ID NO:391 and SEQ ID NO:392; SEQ ID NO:391 and SEQ ID NO:393; SEQ ID NO:391 and SEQ ID NO:394; SEQ ID NO:391 and SEQ ID NO:395; SEQ ID NO:391 and SEQ ID NO:396; SEQ ID NO:391 and SEQ ID NO:397; SEQ ID SEQ ID NO:391 and SEQ ID NO:398; SEQ ID NO:391 and SEQ ID NO:399; SEQ ID NO:391 and SEQ ID NO:400; SEQ ID NO:391 and SEQ ID NO:401; SEQ ID NO:391 and SEQ ID NO:402; SEQ ID NO:391 and SEQ ID NO:403; SEQ ID NO:391 and SEQ ID NO:404; SEQ ID NO:391 and SEQ ID NO:405; SEQ ID NO:391 and SEQ ID NO:406; SEQ ID NO:391 and SEQ ID NO:407; SEQ ID NO:391 and SEQ ID NO:408; SEQ ID NO:391 and SEQ ID NO:409; SEQ ID NO:391 and SEQ ID NO:410; SEQ ID NO:391 and SEQ ID NO:411; SEQ ID NO:391 and SEQ ID NO:412; SEQ ID NO:391 and SEQ ID NO:413; SEQ ID NO:391 and SEQ ID NO:414; SEQ ID NO:391 and SEQ ID NO: 415; SEQ ID NO:391 and SEQ ID NO:416.

[0160] In some embodiments, the siRNA comprises a modification pattern selected from the group consisting of: SEQ ID NO: 380 and SEQ ID NO: 392; SEQ ID NO: 380 and SEQ ID NO: 393; SEQ ID NO: 381 and SEQ ID NO: 393; SEQ ID NO: 382 and SEQ ID NO: 392; SEQ ID NO: 383 and SEQ ID NO: 394; SEQ ID NO: 383 and SEQ ID NO: 395; SEQ ID NO: 384 and SEQ ID NO: 395; SEQ ID NO: 385 and SEQ ID NO: 396; SEQ ID NO: 386 and SEQ ID NO: 397; SEQ ID NO: 386 and SEQ ID NO: 398; SEQ ID NO: 386 and SEQ ID NO: 399; SEQ ID NO: 386 and SEQ ID NO: 400; SEQ ID NO: 386 and SEQ ID NO: 401; SEQ ID NO: 386 and SEQ ID NO: 402. NO:386 and SEQ ID NO:402; SEQ ID NO:386 and SEQ ID NO:403; SEQ ID NO:386 and SEQ ID NO:404; SEQ ID NO:386 and SEQ ID NO:405; SEQ ID NO:386 and SEQ ID NO:406; SEQ ID NO:387 and SEQ ID NO:395; SEQ ID NO:387 and SEQ ID NO:394; SEQ ID NO:388 and SEQ ID NO:403; SEQ ID NO:388 and SEQ ID NO:399; SEQ ID NO:388 and SEQ ID NO:407; SEQ ID NO:388 and SEQ ID NO:406; SEQ ID NO:388 and SEQ ID NO:415; SEQ ID NO:388 and SEQ ID NO:416; SEQ ID NO:388 and SEQ ID NO:404; SEQ ID NO:388 and SEQ ID NO:402; SEQ ID NO:388 and SEQ ID NO:411; SEQ ID NO:389 and SEQ ID NO:408; SEQ ID NO:389 and SEQ ID NO:412; SEQ ID NO:390 and SEQ ID NO:409; SEQ ID NO:390 and SEQ ID NO:413;SEQ ID NO: 391 and SEQ ID NO: 410; SEQ ID NO: 391 and SEQ ID NO: 414.;

[0161] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 8, 9, 12, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, and 11 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0162] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 8, 9, 12, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 9, 11, and 13 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0163] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 9, 12, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 9, 11, and 13 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0164] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 4, 12, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, and 11 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0165] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 8, 9, 10, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 15 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0166] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 8, 9, 10, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, and 10 at the 5' end of the sense strand are 2'-fluoro nucleotides, position 11 is a deoxy nucleotide, and the remaining positions are 2'-methoxy nucleotides.

[0167] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 4, 6, 8, 14, 16, 18, and 20 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, and 10 at the 5' end of the sense strand are 2'-fluoro nucleotides, position 11 is a deoxy nucleotide, and the remaining positions are 2'-methoxy nucleotides.

[0168] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 14, 16, and 18 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 13 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0169] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 8, 9, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 16 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0170] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 8, 9, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 17 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0171] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 8, 9, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 11, and 17 at the 5' end of the sense strand are 2'-fluoro nucleotides, position 10 is a deoxy nucleotide, and the remaining positions are 2'-methoxy nucleotides.

[0172] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 8, 9, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 10, 11, and 17 at the 5' end of the sense strand are 2'-fluoro nucleotides, position 9 is a deoxy nucleotide, and the remaining positions are 2'-methoxy nucleotides.

[0173] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 8, 9, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, and 17 at the 5' end of the sense strand are 2'-fluoro nucleotides, position 11 is a deoxy nucleotide, and the remaining positions are 2'-methoxy nucleotides.

[0174] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 8, 9, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 9, 10, 11, and 17 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0175] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 8, 9, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, 16, and 17 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0176] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 8, 9, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, 15, and 17 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0177] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 8, 9, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, and 11 at the 5' end of the sense strand are 2'-fluoro nucleotides, position 10 is a deoxy nucleotide, and the remaining positions are 2'-methoxy nucleotides.

[0178] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 4, 6, 8, 10, 14, 16, 18, and 20 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 15 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0179] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 4, 6, 8, 10, 14, 16, 18, and 20 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, and 10 at the 5' end of the sense strand are 2'-fluoro nucleotides, position 11 is a deoxy nucleotide, and the remaining positions are 2'-methoxy nucleotides.

[0180] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 8, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, and 11 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0181] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 8, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, and 10 at the 5' end of the sense strand are 2'-fluoro nucleotides, position 11 is a deoxy nucleotide, and the remaining positions are 2'-methoxy nucleotides.

[0182] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 8, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 13 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0183] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 8, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 14 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0184] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 8, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 15 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0185] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 8, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 17 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0186] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 8, 14, and 16 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 19 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0187] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 14, 16, and 18 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, and 11 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0188] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 14, 16, and 18 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, and 10 at the 5' end of the sense strand are 2'-fluoro nucleotides, position 11 is a deoxy nucleotide, and the remaining positions are 2'-methoxy nucleotides.

[0189] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 14, 16, and 18 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 14 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0190] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 14, 16, and 18 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 15 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0191] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 14, 16, and 18 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 17 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0192] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 14, 16, and 18 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 19 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0193] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 14, 16, and 20 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, and 11 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0194] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 14, 16, and 20 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, and 10 at the 5' end of the sense strand are 2'-fluoro nucleotides, position 11 is a deoxy nucleotide, and the remaining positions are 2'-methoxy nucleotides.

[0195] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 14, 16, and 20 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 13 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0196] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 14, 16, and 20 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 14 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0197] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 14, 16, and 20 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 15 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0198] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 14, 16, and 20 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 17 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0199] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length, wherein positions 2, 6, 14, 16, and 20 at the 5' end of the antisense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides; the sense strand of the siRNA is 21 nucleotides in length, wherein positions 7, 9, 10, 11, and 19 at the 5' end of the sense strand are 2'-fluoro nucleotides, and the remaining positions are 2'-methoxy nucleotides.

[0200] In some embodiments of the present invention, the siRNA of the present invention comprises a modified internucleoside linkage or a modified backbone. The modified internucleoside linkage or backbone includes, but is not limited to, phosphorothioate, 2'-O methoxyethyl (MOE), 2'-fluoro, alkyl phosphate, phosphorodithioate, alkyl phosphorothioate, phosphoramidate, carbamate, carbonate, phosphotriester, acetamidate, carboxymethyl ester, and combinations thereof.

[0201] In some embodiments of the present invention, the modified nucleotide is a nucleotide in which the phosphate group is modified with a phosphorothioate group, that is, a non-bridging oxygen atom in the phosphodiester bond is replaced with a sulfur atom, thereby replacing the phosphodiester bond with a phosphorothioate diester bond.

[0202] In some embodiments of the invention, the 5' and 3' ends of the sense strand independently comprise 1 or 2 phosphorothioate linkages; and / or the 5' and 3' ends of the antisense strand independently comprise 1 or 2 phosphorothioate linkages.

[0203] In some embodiments of the present invention, at least one of the following linkages is phosphorothioate between the 1st and 2nd nucleotides at the 5' end of the sense strand, between the 2nd and 3rd nucleotides at the 5' end of the sense strand, between the 1st and 2nd nucleotides at the 3' end of the sense strand, between the 2nd and 3rd nucleotides at the 3' end of the sense strand, between the 1st and 2nd nucleotides at the 3' end of the antisense strand, between the 2nd and 3rd nucleotides at the 3' end of the antisense strand, between the 1st and 2nd nucleotides at the 5' end of the antisense strand, and between the 2nd and 3rd nucleotides at the 5' end of the antisense strand; in some embodiments of the present invention, at least four of the linkages are phosphorothioate; in some embodiments of the present invention, at least six of the linkages are phosphorothioate; in some embodiments of the present invention, all eight of the linkages are phosphorothioate.

[0204] In some embodiments of the present invention, the nucleotides at positions 1 and 2, and the nucleotides at positions 2 and 3 at the 5' end of the sense strand are linked by phosphorothioate groups.

[0205] In some embodiments of the present invention, the nucleotides at positions 1 and 2, and 2 and 3 of the sense strand are linked by phosphorothioate groups, and the nucleotides at positions 1 and 2 of the sense strand are linked by phosphorothioate groups.

[0206] In some embodiments of the present invention, the nucleotides at positions 1 and 2 and the nucleotides at positions 2 and 3 of the 5' end of the sense strand are linked by phosphorothioate groups, and the nucleotides at positions 1 and 2 and the nucleotides at positions 2 and 3 of the 3' end are linked by phosphorothioate groups.

[0207] In some embodiments of the present invention, the nucleotides at positions 1 and 2 and the nucleotides at positions 2 and 3 at the 3' end of the antisense strand are linked by phosphorothioate groups, and the nucleotides at positions 1 and 2 and the nucleotides at positions 2 and 3 at the 5' end are linked by phosphorothioate groups.

[0208] In some embodiments of the present invention, the nucleotides between the 1st and 2nd positions at the 5' end of the sense chain, the nucleotides between the 2nd and 3rd positions at the 5' end of the sense chain, the nucleotides between the 1st and 2nd positions at the 3' end of the antisense chain, the nucleotides between the 2nd and 3rd positions at the 3' end of the antisense chain, the nucleotides between the 1st and 2nd positions at the 5' end of the antisense chain, and the nucleotides between the 2nd and 3rd positions at the 5' end of the antisense chain are all linked by phosphorothioate groups.

[0209] In some embodiments of the present invention, the nucleotides between the 1st and 2nd positions at the 5' end of the sense chain, between the nucleotides between the 2nd and 3rd positions at the 5' end of the sense chain, between the nucleotides between the 1st and 2nd positions at the 3' end of the sense chain, between the nucleotides between the 1st and 2nd positions at the 3' end of the antisense chain, between the nucleotides between the 2nd and 3rd positions at the 3' end of the antisense chain, between the nucleotides between the 1st and 2nd positions at the 5' end of the antisense chain, and between the nucleotides between the 2nd and 3rd positions at the 5' end of the antisense chain are all linked by phosphorothioate groups.

[0210] In some embodiments of the present invention, the nucleotides between the 1st and 2nd positions at the 5' end of the sense chain, the nucleotides between the 2nd and 3rd positions at the 5' end of the sense chain, the nucleotides between the 1st and 2nd positions at the 3' end of the sense chain, the nucleotides between the 2nd and 3rd positions at the 3' end of the sense chain, the nucleotides between the 1st and 2nd positions at the 3' end of the antisense chain, the nucleotides between the 2nd and 3rd positions at the 3' end of the antisense chain, the nucleotides between the 1st and 2nd positions at the 5' end of the antisense chain, and the nucleotides between the 2nd and 3rd positions at the 5' end of the antisense chain are all linked by phosphorothioate groups.

[0211] In some embodiments of the present invention, the sense strand may include one or more blocking residues or moieties, referred to as "blocking residues." A "blocking residue" is a non-nucleotide compound or other moiety that can be incorporated into one or more ends of the nucleotide sequence of the siRNA. In some embodiments of the present invention, the blocking residue is present at the 5' end, the 3' end, or both the 5' end and the 3' end of the sense strand.

[0212] In some embodiments of the present invention, an inverted abasic residue (iab) is added as a capping residue. See F. Czauderna, Nucleic Acids Res., 2003, 31(11), 2705-16. In some embodiments of the present invention, the 5' end and / or 3' end of the sense strand may contain more than one inverted abasic deoxyribose moiety as a capping residue.

[0213] In some embodiments of the invention, one or more inverted abasic residues (iab) are added to the 3' end of the sense strand. In some embodiments of the invention, one or more inverted abasic residues (iab) are added to the 5' end of the sense strand. In some embodiments of the invention, one or more inverted abasic residues are included at or near one or more ends of the siRNA sense strand.

[0214] Inverted abasic residues can be linked via phosphate, phosphorothioate, or other internucleoside linkages.

[0215] In some embodiments of the present invention, the first nucleotide at the 5' end of the antisense strand comprises a (E)-vinylphosphonate and its analogs modification, such as a (E)-VP modification. In some embodiments of the present invention, the first nucleotide may optionally comprise other modifications, such as a 2'-modification (e.g., a 2'-methoxy modification).

[0216] In some embodiments of the present invention, the nucleotides 2 to 8 at the 5' end of the antisense strand are nucleotides modified with glycerol (GNA) and its analogs.

[0217] In some embodiments, the antisense strand comprises the nucleotide sequence shown in any one of SEQ ID NOs: 249-302.

[0218] In some embodiments, the sense strand comprises the nucleotide sequence shown in any one of SEQ ID NOs: 303-379.

[0219] In some embodiments, the sequences of the sense and antisense strands of the siRNA are selected from the sense and antisense strand sequences of any duplex 125 to duplex 172, duplex 177, duplex 180 to duplex 185, and duplex 187 to duplex 237 provided in Table 2.

[0220] In some embodiments, the antisense strand comprises the nucleotide sequence shown in any one of SEQ ID NOs: 256, 260, 263, 264, 278, 279, 281, 285, 286, 287, and 288. In some embodiments, the sense strand comprises the nucleotide sequence shown in any one of SEQ ID NOs: 310, 313, 314, 315, 318, 317, 321, 324, 325, 326, 329, 330, 331, 333, 334, 349, 350, 351, 352, 356, 357, 358, 359, 360, 361, 362, 363, 364, and 365. In some embodiments, the sequences of the sense and antisense strands of the siRNA are selected from the sense and antisense strand sequences of any one of duplex 143, duplex 144, duplex 145, duplex 146, duplex 147, duplex 148, duplex 153, duplex 157, duplex 158, duplex 167, duplex 168, duplex 169, duplex 170, duplex 171, duplex 172, duplex 198, duplex 200, duplex 202, duplex 203, duplex 208, duplex 209, duplex 210, duplex 211, duplex 212, duplex 213, duplex 214, duplex 215, duplex 216, and duplex 217 provided in Table 2.

[0221] In some embodiments, the antisense strand comprises the nucleotide sequence shown in any one of SEQ ID NOs: 256, 264, 285, 286, 287, and 288. In some embodiments, the sense strand comprises the nucleotide sequence shown in any one of SEQ ID NOs: 310, 314, 313, 318, 317, 330, 331, 334, 356, 357, 358, 359, 360, 361, 362, 363, 364, and 365. In some embodiments, the sequences of the sense and antisense strands of the siRNA are selected from the sense and antisense strand sequences of any one of duplex 143, duplex 144, duplex 145, duplex 146, duplex 148, duplex 167, duplex 169, duplex 171, duplex 208, duplex 209, duplex 210, duplex 211, duplex 212, duplex 213, duplex 214, duplex 215, duplex 216, and duplex 217 provided in Table 2.

[0222] On the other hand, the present invention also provides a small interfering RNA (siRNA), wherein the siRNA comprises a sense strand and an antisense strand, wherein the antisense strand comprises a modification pattern selected from the following 1) to 12):

[0223] 1)AS(5'-3'):mN*fN*mNmNmNfNmNfNfNmNmNmNmNfNmNfNmNmNmNmNmNmN*mN*mN(SEQ ID NO:380);

[0224] 2)AS(5'-3'):mN*fN*mNmNmNmNmNmNfNmNmNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN(SEQ ID NO:381);

[0225] 3)AS(5'-3'):mN*fN*mNfNmNmNmNmNmNmNmNmNmNfNmNfNmNmNmNmNmNmN*mN*mN(SEQ ID NO:382);

[0226] 4)AS(5'-3'):mN*fN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNfNmNfNmNmNmNmNmN*m N*mN(SEQ ID NO:383);

[0227] 5)AS(5'-3'):mN*fN*mNfNmNfNmNfNmNmNmNmNmNmNmNfNmNfNmNfNmNmN*mN*mN(SEQ ID NO:384);

[0228] 6)AS(5'-3'):mN*fN*mNmNmNmNfNmNmNmNmNmNmNmNmNmNmNfNmNfNmNmNmNmNmNmN*mN*mN(SEQ ID NO:385);

[0229] 7)AS(5'-3'):mN*fN*mNmNmNmNmNfNfNmNmNmNmNmNmNmNfNfNmNmNmNmNmNmN*mN*mN(SEQ ID NO:386);

[0230] 8)AS(5'-3'):mN*fN*mNfNmNfNmNfNmNfNmNmNmNmNmNfNmNfNfNfNmNmN*mN*mN(SEQ ID NO:387);

[0231] 9)AS(5'-3'):(E)-VPmN*fN*mNmNmNmNmNfNfNmNmNmNmNmNfNfNfNmNmNmNmNmN*mN*mN(SEQ ID NO:388);

[0232] 10)AS(5'-3'):(E)-VPmN*fN*mNmNmNfNmNfNfNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 389);

[0233] 11)AS(5'-3'):(E)-VPmN*fN*mNmNmNmNmNmNfNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 390); or,

[0234] 12)AS(5'-3'):(E)-VPmN*fN*mNmNmNfNmNfNfNfNmNmNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 391);

[0235] Wherein, mN is a methoxy-modified nucleotide, fN is a fluorinated-modified nucleotide, (E)-VP indicates that the nucleotide adjacent to its right is a (E)-vinylphosphonate-modified nucleotide, and * indicates a phosphorothioate linkage;

[0236] and / or,

[0237] The sense strand comprises a modification pattern selected from the following 1) to 25):

[0238] 1) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmNmNmN (SEQ ID NO: 392);

[0239] 2) SS(5'-3'):mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmN (SEQ ID NO: 393);

[0240] 3) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNmNmNmNmNmNmN (SEQ ID NO: 394);

[0241] 4) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmNmN (SEQ ID NO: 395);

[0242] 5) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNfNmNmNmNmNmNmNmNm N (SEQ ID NO: 396);

[0243] 6)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmN(SEQ ID NO:397);

[0244] 7)SS(5'-3'):mN*mN*mNmNmNmNmNfNfNfNfNmNmNmNmNmNmNfNfNfNmNmNmNmNmNmNmNmNmNmNmN(SEQ ID NO:398);

[0245] 8)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNfNmNmNmNmN*mN(SEQ ID NO:399);

[0246] 9)SS(5'-3'):mN*mN*mNmNmNmNfNmNdNfNfNfNmNmNmNmNmNfNmNmNmNmN*mN*mN(SEQ ID NO:400);

[0247] 10)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNfNmNmNmNmN*mN(SEQ ID NO:401);

[0248] 11)SS(5'-3'):mN*mN*mNmNmNmNmNmNfNfNfNfNmNmNmNmNmNfNfNmNmNmN*mN(SEQ ID NO:402);

[0249] 12)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNfNmNmNmNmN*mN(SEQ ID NO:403);

[0250] 13)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNfNmNmNmNmNmNfNmNmN*mN*mN(SEQ ID NO:404);

[0251] 14)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNfNmNmNmNmNfNfNmNmNmNmN*mN(SEQ ID NO:405);

[0252] 15)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNfNfNmNmNmN*mN(SEQ ID NO:406);

[0253] 16) SS(5’-3’): mN*mN*mNmNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmN*mN (SEQ ID NO: 407);

[0254] 17) SS(5’-3’): mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmN*mN*mN (SEQ ID NO: 408);

[0255] 18) SS(5’-3’): mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmN*mN*mN (SEQ ID NO: 409);

[0256] 19) SS(5’-3’): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmN*mN*mN (SEQ ID NO: 410);

[0257] 20) SS(5’-3’): mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmN*mN *mN (SEQ ID NO: 411);

[0258] 21) SS(5’-3’): mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmNmNmN*(iab) (SEQ ID NO: 412);

[0259] 22) SS(5’-3’): mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmN*(iab) (SEQ ID NO: 413);

[0260] 23) SS(5’-3’): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmNmN*(iab) (SEQ ID NO: 414);

[0261] 24) SS(5’-3’): mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmNmNmN*(iab) (SEQ ID NO: 415); or,

[0262] 25)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNfNmNmNmNmN*(iab) (SEQ ID NO: 416);

[0263] Wherein, mN is a methoxy-modified nucleotide, fN is a fluorinated-modified nucleotide, dN is a deoxynucleotide, * is a phosphorothioate linkage, and (iab) is an inverted abasic residue.

[0264] In some embodiments, the siRNA comprises a modification pattern selected from the group consisting of: SEQ ID NO: 380 and SEQ ID NO: 392; SEQ ID NO: 380 and SEQ ID NO: 393; SEQ ID NO: 381 and SEQ ID NO: 393; SEQ ID NO: 382 and SEQ ID NO: 392; SEQ ID NO: 383 and SEQ ID NO: 394; SEQ ID NO: 383 and SEQ ID NO: 395; SEQ ID NO: 384 and SEQ ID NO: 395; SEQ ID NO: 385 and SEQ ID NO: 396; SEQ ID NO: 386 and SEQ ID NO: 397; SEQ ID NO: 386 and SEQ ID NO: 398; SEQ ID NO: 386 and SEQ ID NO: 399; SEQ ID NO: 386 and SEQ ID NO: 400; SEQ ID NO: 386 and SEQ ID NO: 401; SEQ ID NO: 386 and SEQ ID NO: 402. NO:386 and SEQ ID NO:402; SEQ ID NO:386 and SEQ ID NO:403; SEQ ID NO:386 and SEQ ID NO:404; SEQ ID NO:386 and SEQ ID NO:405; SEQ ID NO:386 and SEQ ID NO:406; SEQ ID NO:387 and SEQ ID NO:395; SEQ ID NO:387 and SEQ ID NO:394; SEQ ID NO:388 and SEQ ID NO:403; SEQ ID NO:388 and SEQ ID NO:399; SEQ ID NO:388 and SEQ ID NO:407; SEQ ID NO:388 and SEQ ID NO:406; SEQ ID NO:388 and SEQ ID NO:415; SEQ ID NO:388 and SEQ ID NO:416; SEQ ID NO:388 and SEQ ID NO:404; SEQ ID NO:388 and SEQ ID NO:402; SEQ ID NO:388 and SEQ ID NO:411; SEQ ID NO:389 and SEQ ID NO:408; SEQ ID NO:389 and SEQ ID NO:412; SEQ ID NO:390 and SEQ ID NO:409; SEQ ID NO:390 and SEQ ID NO:413;SEQ ID NO: 391 and SEQ ID NO: 410; SEQ ID NO: 391 and SEQ ID NO: 414.;

[0265] deliver

[0266] The siRNA of the present invention can be delivered or introduced by any means known in the art (e.g., delivered in vitro or introduced into cells, or delivered in vivo or introduced into patients). For example, to deliver in vivo, the siRNA can be injected into a tissue site, or systemically administered. In vivo delivery can also be carried out by a beta-glucan delivery system. In vitro introduction into cells includes methods known in the art, such as electroporation and lipofection.

[0267] In some embodiments, the delivery method includes, but is not limited to, viral delivery (retrovirus, adenovirus, lentivirus, baculovirus, AAV); liposomes (Lipofectamine, cationic DOTAP, neutral DOPC); nanoparticles (cationic polymers, PEI); bacterial delivery (tkRNAi); chemical modification of siRNA (LNA) to increase stability; lipid nanoparticles (LNP); neutral liposomes (NL); polymer nanoparticles (low molecular weight polymers or high molecular weight polymers); double-stranded RNA binding motifs (dsRBMs); and other delivery systems known in the art to be suitable for nucleic acid or oligonucleotide delivery.

[0268] Conjugate

[0269] In one aspect, the present invention provides a conjugate comprising at least one siRNA of the present invention and a pharmaceutically acceptable targeting molecule. The conjugate of the present invention is obtained by coupling the siRNA of the present invention to a pharmaceutically acceptable targeting molecule, the conjugate comprising a pharmaceutically acceptable targeting molecule and an optional linker. The siRNA can be non-covalently coupled to the targeting molecule, or covalently coupled to the targeting molecule.

[0270] The pharmaceutically acceptable targeting molecule can be a targeting molecule conventionally used in the field of siRNA administration, which typically enhances the pharmacokinetic or biodistribution properties of the siRNA to which it is attached, and improves the cell-specific (or organ-specific) distribution and cell-specific (or organ-specific) uptake of the siRNA. Representative targeting molecules include, but are not limited to, compounds with affinity for cell surface molecules, cell receptor ligands, haptens, antibodies or antibody fragments, antibody mimics, and the like. In some embodiments, the targeting molecules include but are not limited to one or more of the following targeting molecules or their derivatives: integrin family; lipophilic molecules, such as cholesterol, bile acid, vitamins (such as vitamin E), lipid molecules of different chain lengths; polymers, such as polyethylene glycol; polypeptides, such as membrane-permeable peptides; aptamers; antibodies; quantum dots; carbohydrates, such as lactose, polylactose, mannose, galactose, N-acetylgalactosamine (GalNAc); folic acid (folate); or receptor ligands expressed by hepatic parenchymal cells, such as asialoglycoproteins, asialosugar residues, lipoproteins (such as high-density lipoproteins, low-density lipoproteins, etc.), glucagon, neurotransmitters (such as adrenaline), growth factors, transferrin, etc.

[0271] The linker can be a linker conventionally used in the field of siRNA administration, including but not limited to one or more of the following linkers or their derivatives: amide linker portion, amino linker portion, carbonyl linker portion, carbamate linker portion, urea linker portion, ether linker portion, disulfide linker portion, succinylamino linker portion, etc.

[0272] In some embodiments, the targeting molecule can be directly or indirectly connected to the siRNA of the present invention via a joint / linking group. In some embodiments, the targeting molecule is connected to the siRNA via an unstable, cleavable or reversible bond or joint. In some embodiments, the targeting molecule is connected to at least one end of the sense strand and / or antisense strand of the siRNA. In some embodiments, the targeting molecule is connected to the 5' end and / or 3' end of the sense strand. In some embodiments, the targeting molecule is connected to the 5' end and / or 3' end of the antisense strand.

[0273] In some embodiments, the siRNA of the present invention is delivered to target cells or tissues via a targeting molecule linked thereto. In some embodiments, the targeting molecule comprises a ligand for a cell receptor.

[0274] In some embodiments, the targeting molecule has affinity for an epithelial cell surface receptor.

[0275] In some embodiments, the targeting molecule is an integrin ligand.

[0276] In some embodiments, the targeting molecule includes an αvβ6 integrin ligand. Integrin αvβ6 is an epithelial-specific integrin that is highly upregulated in damaged lung epithelial cells. Herein, "αvβ6 integrin ligand" refers to a molecule with affinity for integrin αvβ6, which can promote the targeting and delivery of siRNA connected thereto to cells expressing integrin αvβ6 (such as lung epithelial cells). αvβ6 integrin ligands are known to those skilled in the art, and can be found in, for example, international patent application WO2023070082A2. In some embodiments, two or more (e.g., 3) αvβ6 integrin ligands are connected to the siRNA of the present invention directly or via a linker / connector group. In some embodiments, the two or more (e.g., 3) αvβ6 integrin ligands are connected to the 5' end of the sense chain.

[0277] Pharmaceutical composition

[0278] In one aspect, the present invention provides pharmaceutical compositions comprising at least one siRNA of the present invention.

[0279] In some embodiments of the present invention, the pharmaceutical composition contains one siRNA as described above.

[0280] In other embodiments of the present invention, the pharmaceutical composition contains at least two of the above-described siRNAs (for example, but not limited to, two, three, four, five, six, seven, eight, nine, ten, or more) as active ingredients. Preferably, the at least two siRNAs each target a different target sequence in the MMP7 gene, thereby enabling simultaneous action against different target sequences and resulting in a synergistic effect. Here, "different target sequences" refers to target sequences that do not overlap, or target sequences that overlap by less than five consecutive nucleotides (for example, the number of overlapping consecutive nucleotides is 4, 3, 2, 1, or 0). In this case, the at least two siRNAs described above may be present in any ratio. Preferably, the at least two siRNAs described above may be present in a molar ratio of 1:100 to 100:1; more preferably, the at least two siRNAs described above may be present in a molar ratio of 1:10 to 10:1, 1:5 to 5:1, or 1:2 to 2:1. In some embodiments of the present invention, the at least two aforementioned siRNAs are present in the same molar ratio.

[0281] In other embodiments of the present invention, the pharmaceutical composition contains at least one siRNA of the present invention and further contains at least one siRNA targeting another target (e.g., another gene other than MMP7). In this case, the siRNA of the present invention and the siRNA targeting the other target can be present in any different ratios, for example, in a molar ratio of 1:100 to 100:1, for example, in a molar ratio of 1:10 to 10:1, 1:5 to 5:1, or 1:2 to 2:1, for example, in the same molar ratio.

[0282] In some embodiments, the pharmaceutical composition comprises an effective amount of siRNA. An "effective amount" refers to an amount of siRNA effective to produce the desired pharmacological, therapeutic, or preventive outcome. For example, if a given clinical treatment is considered effective when there is at least a 10% reduction in a measurable parameter associated with a disease or condition, then a therapeutically effective amount of the drug for treating that disease or condition is the amount required to achieve at least a 10% reduction in that parameter. For example, a therapeutically effective amount of an siRNA targeting MMP7 can reduce MMP7 mRNA levels by at least 10%.

[0283] In some embodiments, in the pharmaceutical composition described in any of the above embodiments, the siRNA can be linked to a targeting molecule to form a conjugate. Therefore, in some embodiments, the pharmaceutical composition comprises a conjugate of the present invention.

[0284] In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or excipient.

[0285] In some embodiments, the pharmaceutically acceptable carrier and / or excipient is a delivery vehicle. A delivery vehicle is a substance that improves the delivery of nucleic acids or oligonucleotides to cells or tissues. Such substances can be any delivery vehicle known in the art that is suitable for nucleic acid or oligonucleotide delivery, including but not limited to: viruses (retroviruses, adenoviruses, lentiviruses, baculoviruses, AAVs); liposomes (Lipofectamine, cationic DOTAP, neutral DOPC); nanoparticles (cationic polymers, PEI); bacteria (tkRNAi); lipid nanoparticles (LNPs); neutral liposomes (NLs); polymer nanoparticles (low molecular weight polymers or high molecular weight polymers); double-stranded RNA binding motifs (dsRBMs), etc.

[0286] In some embodiments, the siRNA can be packaged by the delivery vehicle.

[0287] In some embodiments, the siRNA can be directly or indirectly connected to the delivery vector via a joint / connecting group. In some embodiments, the delivery vector is connected to the siRNA via an unstable, cleavable or reversible bond or joint. In some embodiments, the delivery vector is connected to at least one end of the sense strand and / or antisense strand of the siRNA. In some embodiments, the delivery vector is connected to the 5' end and / or 3' end of the sense strand. In some embodiments, the delivery vector is connected to the 5' end and / or 3' end of the antisense strand.

[0288] In some embodiments, the pharmaceutical composition of the present invention is formulated into a dosage form compatible with its intended route of administration, for example, it can be administered locally (such as direct injection or implantation), systemically, or subcutaneously, intravenously, intraperitoneally, or parenterally, including intracranial (such as intraventricular, intrameningeal, or intrathecal), intramuscularly, transdermally, airway (aerosol), nasal, oral, rectal, or topical (including buccal and sublingual) administration.

[0289] In some embodiments, the pharmaceutical composition is administered by suction, intranasal administration, intratracheal administration or oropharyngeal inhalation. The active ingredient of the required amount can be mixed in a suitable solvent and then aseptically filtered to prepare a preparation suitable for inhalation administration. Generally, the preparation for inhalation administration is a sterile solution under physiological pH and has low viscosity. Salt can be added to the preparation to balance tension. In some cases, a surfactant or cosolvent can be added to increase the solubility of the active ingredient and improve aerosol properties. In some cases, an excipient can be added to control viscosity to ensure the size and distribution of the atomized droplets.

[0290] In other embodiments, the pharmaceutical composition can be administered by injection, such as intravenous, intramuscular, subcutaneous, intradermal, intraarticular, intraocular, intraperitoneal or topical administration. Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the temporary preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, or phosphate buffered saline (PBS) and the like. The pharmaceutical composition should remain stable under production and storage conditions and should prevent contamination by microorganisms such as bacteria and fungi. For example, a sterile injection solution can be prepared by the following method: the active ingredient of the required dose is mixed in an appropriate solvent, and optionally, other desired ingredients (including but not limited to, pH regulators, surfactants, adjuvants, ionic strength enhancers, diluents, agents for maintaining osmotic pressure, agents for delayed absorption, preservatives, or any combination thereof) are mixed at the same time, followed by filtration sterilization. In addition, the sterile injection solution can be prepared as a sterile lyophilized powder (for example, by vacuum drying or freeze drying) for easy storage and use.

[0291] The siRNA of the present invention can be formulated in dosage unit form for ease of administration. Dosage unit form refers to physically discrete units suitable as unitary dosages for the subject to be treated; each unit contains a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.

[0292] In the present invention, the dosage regimen can be adjusted to obtain the best desired response (e.g., therapeutic or preventive response). For example, the dosage can be a single dose, multiple doses can be administered over a period of time, or the dosage can be proportionally reduced or increased according to the urgency of the treatment situation.

[0293] application

[0294] Inhibits the expression of MMP7

[0295] The siRNA of the present invention can be used to inhibit the expression of MMP7 in vitro and / or in vivo.

[0296] In one aspect, the present invention provides a method for inhibiting MMP7 expression in a cell, comprising introducing into the cell an siRNA, conjugate, or pharmaceutical composition of the present invention. In some embodiments, the method is performed in vitro. The siRNA of the present invention can be introduced by any nucleic acid delivery method known in the art, such as electroporation or lipofection.

[0297] The term "inhibiting MMP7 expression" refers to at least partial suppression of MMP7 gene expression, which can be manifested as a decrease in the amount of detectable MMP7 mRNA. The degree of inhibition is typically expressed as: (mRNA in control cells) - (mRNA in treated cells) / (mRNA in control cells) * 100%. Alternatively, the degree of inhibition can be given as a decrease in a parameter functionally associated with MMP7 gene expression, such as the amount of protein encoded by the MMP7 gene. In principle, MMP7 gene silencing can be measured in any cell expressing MMP7 (constitutively or genetically engineered) and by any suitable assay. Measurements can be made at multiple time points, before, during, and after administration of the siRNA, to determine the effect of the siRNA. MMP7 levels or expression can be measured by evaluating mRNA (e.g., by Northern blot or PCR) or protein (e.g., by Western blot or ELISA). For example, the effect of the siRNA on MMP7 expression can be determined by measuring the rate of MMP7 gene transcription (e.g., by RT-PCR).

[0298] In some embodiments, the expression of the MMP7 gene is suppressed by at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% by administering the siRNA of the present invention. In some embodiments, the expression of the MMP7 gene is suppressed by at least about 60%, 70%, or 80% by administering the siRNA of the present invention. In some embodiments, the expression of the MMP7 gene is suppressed by at least about 85%, 90%, or 95% by administering the siRNA of the present invention. In some embodiments, the expression of the MMP7 gene is suppressed by at least about 96%, 97%, 98%, 99%, or 100% by administering the siRNA of the present invention.

[0299] In some embodiments, the siRNA, the conjugate, or the pharmaceutical composition is used alone or in combination with another pharmaceutically active agent (eg, siRNA targeting a different target sequence in the MMP7 gene or siRNA targeting other targets).

[0300] In some embodiments, one siRNA provided herein is used. In other embodiments, at least two siRNAs provided herein (for example, but not limited to, two, three, four, five, six, seven, eight, nine, ten, or more) are used, preferably each of the at least two siRNAs targets a different target sequence in the MMP7 gene. In other embodiments, at least one siRNA provided herein and an siRNA targeting other targets (e.g., genes other than MMP7) are used.

[0301] Treatment of MMP7-related diseases

[0302] The siRNAs of the present invention can be used to treat diseases or conditions that would benefit from reduced levels or inhibition of expression of MMP7.

[0303] In one aspect, the present invention provides a method for preventing and / or treating a pathological condition or disease associated with MMP7 in a subject, the method comprising administering to a subject in need thereof an effective amount of an siRNA, conjugate, or pharmaceutical composition of the present invention. The present invention also relates to the use of an siRNA, conjugate, or pharmaceutical composition of the present invention in the preparation of a medicament for treating and / or preventing a pathological condition or disease associated with MMP7.

[0304] In some embodiments, the pathological condition or disease associated with MMP7 involves overexpression of MMP7. Overexpression of MMP7 refers to MMP7 levels (e.g., MMP7 levels present in plasma or tissues of a subject, and preferably in damaged tissues) that are higher than normal MMP7 levels (e.g., corresponding levels in healthy controls).

[0305] In some embodiments, the pathological condition or disease associated with MMP7 would benefit from reduced levels or inhibition of MMP7 expression.

[0306] In some embodiments, the pathological condition or disease associated with MMP7 is an inflammatory disease of the lung, such as pulmonary fibrosis (eg, idiopathic pulmonary fibrosis, interstitial lung disease) or chronic obstructive pulmonary disease.

[0307] In some embodiments, the pathological condition or disease associated with MMP7 is a fibrotic disease, such as renal fibrosis or liver fibrosis.

[0308] In some embodiments, the pathological condition or disease associated with MMP7 is idiopathic pulmonary fibrosis.

[0309] In some embodiments, if MMP7 expression is elevated for a particular disease, treatment with the siRNA of the invention may preferably reduce the level or expression of MMP7 to a level that is within the range considered normal for individuals without such a disorder.

[0310] In some embodiments, the subject is a mammal, such as a human.

[0311] In some embodiments, the siRNA, the conjugate or the pharmaceutical composition is used alone or in combination with another pharmaceutically active agent (e.g., siRNA targeting a different target sequence in the MMP7 gene or siRNA targeting other targets), for example, administered simultaneously or sequentially.

[0312] In some embodiments, one siRNA provided herein is used. In other embodiments, at least two siRNAs provided herein (for example, but not limited to, two, three, four, five, six, seven, eight, nine, ten, or more) are used, preferably each of the at least two siRNAs targets a different target sequence in the MMP7 gene. In other embodiments, at least one siRNA provided herein and an siRNA targeting other targets (e.g., genes other than MMP7) are used.

[0313] In some embodiments, the siRNA of the invention is administered by inhalation, intranasally, intratracheally, or oropharyngeal inhalation. In some embodiments, the siRNA of the invention is administered by inhalation, for example, by an inhaler device, such as a metered-dose inhaler, or a nebulizer such as a jet-nebulizer or a vibrating mesh nebulizer, or a soft mist inhaler, for the treatment of inflammatory lung diseases.

[0314] Definition of terms

[0315] In the present invention, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. At the same time, in order to better understand the present invention, the definitions and explanations of relevant terms are provided below.

[0316] In this document, unless otherwise specified, capital letters C, G, U, A, and T represent the base composition of nucleotides, including modified and unmodified nucleotides; lowercase letter m indicates that the nucleotide adjacent to the right of the identifier m is a 2'-methoxy nucleotide; lowercase letter f indicates that the nucleotide adjacent to the right of the identifier f is a 2'-fluoro nucleotide; lowercase letter d indicates that the nucleotide adjacent to the right of the identifier d is a 2'-deoxy nucleotide; the identifier * indicates that the two nucleotides adjacent to the left and right of the identifier * are connected by thiophosphate groups; (E)-VP indicates that the nucleotide adjacent to the right is a (E)-vinylphosphonate-modified nucleotide; and iab indicates an inverted abasic residue.

[0317] As used herein, the term "modified nucleotide" refers to a nucleotide independently having a modified ribose moiety, a modified internucleoside bond, or a modified base. Thus, the term "modified nucleotide" encompasses substitutions, additions, or removals (e.g., using functional groups or atoms) of internucleoside bonds, ribose moieties, or bases. Modifications suitable for use in the present invention include all types of modifications disclosed herein or known in the art. "Methoxy-modified nucleotides" refer to nucleotides in which the 2'-hydroxyl group of the ribose group of a nucleotide is replaced by a methoxy group. "Fluoro-modified nucleotides" refer to nucleotides in which the 2'-hydroxyl group of the ribose group of a nucleotide is replaced by a fluorine group. "Nucleotide analogs" refer to groups that can replace nucleotides in nucleic acids but have a structure different from adenine ribonucleotides, guanine ribonucleotides, cytosine ribonucleotides, uracil ribonucleotides, or thymine deoxyribonucleotides. Such as isonucleotides, bridged nucleic acids (BNAs), or acyclic nucleotides.

[0318] Among them, the structure of the fluorinated modified nucleotide is as follows:

[0319] The structure of a methoxy-modified nucleotide is shown below:

[0320] The nucleotide structure of the locked nucleic acid modification is shown below:

[0321] As used herein, the term "siRNA" refers to an RNA molecule that is capable of sequence-specifically inducing RNAi, comprises a sense strand and an antisense strand, and has a partially or completely complementary double-stranded structure. In the siRNAs of the present invention, the length of the complementary double-stranded structure can be 17-30 base pairs, for example, 19, 20, 21, 22, 23, 24, or 25 base pairs. In some embodiments of the present invention, the siRNA may further contain modified nucleotides as needed, provided that the modified nucleotides do not significantly weaken or abolish the siRNA's ability to inhibit MMP7 gene expression. Currently, there are various methods available in the art for modifying siRNA, including, for example, backbone modifications (e.g., phosphate group modifications), ribose group modifications, and base modifications (Watts, JK, GF Deleavey, and MJ Damha, Chemically modified siRNA: tools and applications. Drug Discov Today, 2008. 13(19-20): p. 842-55).

[0322] The term "antisense strand" includes a region that is substantially complementary to the target sequence. The term "sense strand," as used herein, refers to a strand that has a region that is substantially complementary to a region of the antisense strand. The term "region of complementarity" refers to a region on the antisense strand that is substantially complementary to MMP7 mRNA or a region on the sense strand that is substantially complementary to the antisense strand. When the region of complementarity is not completely complementary to the target sequence or antisense strand, mismatches may occur within or at the terminal regions of the molecule. Typically, the most tolerated mismatches are in the terminal regions, for example, within 5, 4, 3, 2, or 1 nucleotides of the 5' and / or 3' termini.

[0323] As used herein, unless otherwise specified, the term "complementary" refers to the ability of an oligonucleotide of a first sequence to hybridize with an oligonucleotide of a second sequence under certain conditions and form a double-stranded structure. "At least partially complementary" means that the two sequences can be completely complementary, or generally have no more than 6, 5, 4, 3, 2, or 1 mismatched base pairings, while retaining the ability to hybridize under relevant conditions. In addition, when two oligonucleotides are designed to form one or more single-stranded overhangs upon hybridization, such overhangs should not be considered mismatches for the purpose of determining complementarity. As used herein, when the above hybridization ability requirements are met, "complementary" sequences may also include or be completely formed from non-Watson-Crick base pairs and / or base pairs formed from non-natural and modified nucleotides. Such non-Watson-Crick base pairs include, but are not limited to, G:U wobble base pairing or Hoogstein base pairing. Correspondingly, in this document, unless otherwise specified, "mismatch" means that the bases at corresponding positions in the siRNA duplex molecule are not paired in a complementary form.

[0324] Those skilled in the art will be able to determine the most appropriate conditions for testing the complementarity of two sequences based on the intended use of the hybridized nucleotides. Such conditions may, for example, be stringent conditions, such as 400 mM NaCl, 40 mM PIPES pH 6.4, 1 mM EDTA, 50°C or 70°C for 12-16 hours, followed by washing. Other conditions, such as physiologically relevant conditions encountered in vivo, may also be used.

[0325] As used herein, unless otherwise specified, "nucleotide sequence differences" refer to changes in the base type of the nucleotides at the same or corresponding positions compared to the original nucleotide sequence. For example, if one nucleotide base in the original nucleotide sequence is A, and the nucleotide base at the same or corresponding position is changed to U, C, G, or dT, dC, dG, etc., a nucleotide sequence difference is considered to exist at that position. It should be noted that if the nucleotides at the same or corresponding positions differ only in the presence or type of modification compared to the original nucleotide sequence, a nucleotide sequence difference is not considered to exist at that position.

[0326] In this article, if not otherwise specified, the term "pharmaceutically acceptable carrier and / or excipient" refers to a carrier, delivery vehicle, diluent, adjuvant and / or the salt / ester / hydrate thereof formed, etc., which are usually chemically or physically compatible with other ingredients (such as siRNA of the present invention) constituting a certain pharmaceutical dosage form, and are physiologically compatible with the subject. "Pharmaceutically acceptable carrier and / or excipient" does not play or is not intended to play a therapeutic effect at the expected dose. Such ingredients can play the following roles: a) help the processing of the drug delivery system during manufacturing, b) protect, support or enhance the stability, bioavailability or patient acceptability of the active ingredient, c) assist product identification, and / or d) enhance the overall safety, effectiveness, delivery, etc. of the active ingredient during storage and use. For example, the siRNA of the present invention can be wrapped by a delivery vehicle. "Pharmaceutically acceptable carriers and / or excipients" include, but are not limited to, viruses, liposomes, nanoparticles, bacteria, lipid nanoparticles (LNPs), neutral liposomes (NLs), polymer nanoparticles, double-stranded RNA binding motifs (dsRBMs), pH adjusters, surfactants, adjuvants, ionic strength enhancers, diluents, agents that maintain osmotic pressure, agents that delay absorption, and preservatives. For example, viruses include, but are not limited to, retroviruses, adenoviruses, lentiviruses, baculoviruses, and AAVs. Liposomes include, but are not limited to, lipofectamine, cationic DOTAP, and neutral DOPC. Nanoparticles include, but are not limited to, cationic polymers and PEI. Bacteria include, but are not limited to, tkRNAi. Polymeric nanoparticles include, but are not limited to, low molecular weight polymers or high molecular weight polymers. pH adjusters include, but are not limited to, phosphate buffers. Surfactants include, but are not limited to, cationic, anionic, or nonionic surfactants, such as Tween-80. Ionic strength enhancers include, but are not limited to, sodium chloride. Preservatives include, but are not limited to, various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, and the like. Agents that maintain osmotic pressure include, but are not limited to, sugars, NaCl, and the like. Agents that delay absorption include, but are not limited to, monostearate and gelatin. Diluents include, but are not limited to, water, aqueous buffers (e.g., buffered saline), alcohols, and polyols (e.g., glycerol).

[0327] As used herein, unless otherwise specified, the term "inhibit" refers to the down-regulation of target gene expression due to siRNA-mediated degradation of target gene mRNA. "Down-regulation" refers to a decrease in target gene expression by 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% or more, or even 100%, relative to the absence of siRNA treatment. A 100% decrease in target gene expression refers to the absence of detectable target gene expression.

[0328] In this article, the term "overhang" or "nucleotide overhang" refers to at least one unpaired nucleotide protruding from the siRNA duplex structure. For example, when the 3' end of one chain of the siRNA extends beyond the 5' end of the other chain (or vice versa), there is an overhang. The siRNA may include an overhang of at least one nucleotide, or the overhang may include at least 2nt, at least 3nt, at least 4nt, at least 5nt or more. The overhang may include nucleotide / nucleoside analogs, or be composed of nucleotide / nucleoside analogs, which include deoxynucleotides / nucleosides. The overhang may be on the sense strand, the antisense strand, or any combination thereof. The nucleotides of the overhang may be present on the 5' end, the 3' end, or both ends of the antisense strand or the sense strand of the siRNA. Accordingly, the term "blunt end" refers to the absence of a nucleotide overhang.

[0329] As used herein, the term "prevention" refers to a method implemented to prevent or delay the occurrence of a disease, condition, or symptom in a subject; and the term "treatment" refers to a method implemented to obtain a beneficial or desired clinical outcome. For the purposes of the present invention, beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms, reduction in the extent of the disease, stabilization (i.e., no longer worsening) of the state of the disease, delaying or slowing the progression of the disease, improvement or alleviation of the state of the disease, and relief of symptoms (whether partial or complete), whether detectable or undetectable. In addition, "treatment" may also refer to prolonging survival compared to the expected survival if not receiving treatment.

[0330] As used herein, the term "effective amount" refers to an amount sufficient to achieve, or at least partially achieve, a desired effect. For example, a prophylactic effective amount is an amount sufficient to prevent, arrest, or delay the onset of a disease; a therapeutic effective amount is an amount sufficient to cure or at least partially arrest the disease and its complications in a patient already suffering from the disease. Determining such an effective amount is well within the capabilities of those skilled in the art. For example, an effective amount for therapeutic use will depend on the severity of the disease being treated, the overall state of the patient's own immune system, the patient's general condition such as age, weight, and sex, the method of administration of the drug, and any other concurrently administered treatments.

[0331] Advantageous Effects of the Invention

[0332] The siRNA of the present invention can effectively inhibit MMP7 gene expression in vitro and / or in vivo, exhibits good stability, negligible cytotoxicity, and immunostimulatory properties, and significantly reduces MMP7 protein levels in animals. Therefore, the siRNA of the present invention can be used to treat diseases or conditions that benefit from reduced MMP7 levels or suppressed expression, and has important clinical value in the treatment of pulmonary inflammation or fibrosis.

[0333] Below in conjunction with embodiment, embodiment of the present invention is described in detail, but those skilled in the art will understand that the following examples are only used to illustrate the present invention, rather than to limit the scope of the invention. According to the following detailed description of preferred embodiments, various objects and advantages of the present invention will become implementable to those skilled in the art.

[0334] Sequence information

[0335] Table 1: Duplex 1 to Duplex 124

[0336] Table 2. Modified duplexes 125 to 237 DETAILED DESCRIPTION

[0337] The invention will now be described in the following non-limiting examples.

[0338] Those skilled in the art will appreciate that the examples are provided to illustrate the present invention by way of example and are not intended to limit the scope of the invention. The experimental methods in the examples are conventional methods unless otherwise specified. Where specific conditions are not specified in the examples, the experiments were carried out according to conventional conditions or the conditions recommended by the manufacturer. Where the manufacturer of the reagents or instruments is not specified, they are all commercially available conventional products.

[0339] It is known to those skilled in the art that the siRNA of the present invention can be obtained by conventional siRNA preparation methods in the art (e.g., solid phase synthesis and liquid phase synthesis), wherein both solid phase synthesis and liquid phase synthesis are commercially available. It is also clear to those skilled in the art that modified nucleotide groups can be introduced into the siRNA of the present invention by using nucleotide monomers with corresponding modifications. Methods for preparing nucleotide monomers with corresponding modifications are well known to those skilled in the art, and commercial monomers are also available on the market.

[0340] Example 1: siRNA synthesis

[0341] siRNA sequences were designed for the MMP7 gene sequence (see NCBI Reference Sequence: NM_002423.5). For the sense and antisense strands of the siRNA sequences of the present invention, as well as the sense and antisense strands of the modified duplexes, deoxynucleoside CPG was used as a solid support. The sense strand was synthesized using the solid support, while the antisense strand was synthesized using universal CPG.

[0342] Sequence synthesis was performed on a 48-channel synthesizer at a 0.2 μmol scale. The phosphoramidite monomer was used at a concentration of 0.05 M, and the activator was 0.3 M BTT.

[0343] The sequence was cut and deprotected in a 1.5 ml tube using AMA in the first step and triethylamine trihydrofluoride to remove the two protecting groups in the second step. For sequences comprising two fully modified sequences, ammoniacal liquor was used for aminolysis. A mixture of acetone: ethanol (80:20 by volume) was used to precipitate the sequence after cutting and deprotection and to dissolve it in RNase-free water. Each sequence was analyzed by LC-MS to determine sequence accuracy, quantitatively measured by spectrophotometer, and purity determined by HPLC.

[0344] After HPLC purification, lyophilization, and quality control, the salt was exchanged using sodium acetate alcohol precipitation and desalted using a 3KD ultrafiltration tube. After desalting, the sense and antisense strands were quantified spectrophotometrically and mixed in a 1:1 ratio and annealed to form siRNA duplexes. The resulting siRNA duplexes are shown in Table 1.

[0345] Example 2: siRNA purification and activity detection

[0346] 2-1. Cell culture and transfection

[0347] A549 cell culture: A549 cells (ATCC No: CCL-185) were cultured to near confluence in F-12K complete medium (Gibco, supplemented with 10% FBS) at 37°C in a 5% CO2 atmosphere. Cells were then trypsinized and plated. 8,000 A549 cells were plated per well of a 96-well plate in 0.1 mL of F-12K medium (Gibco, supplemented with 10% FBS) and cultured for 24 hours. siRNA was transfected using lipofectamine 2000 (Invitrogen) and cultured for an additional 24 hours at 37°C in a 5% CO2 atmosphere before RNA extraction. Single-dose experiments were performed at duplex concentrations of 10 nM, 1.0 nM, 0.3 nM, 0.1 nM, 0.03 nM, and 0.01 nM (two or three concentrations were tested).

[0348] RNA extraction

[0349] An RNA extraction kit (Yeasen, Cat: 18600ES50) was used according to the instructions of the RNA extraction kit. Finally, 170 μL of RNase-free water was added to collect RNA.

[0350] 2-3. Real-time fluorescence quantitative PCR

[0351] One-step RT-qPCR was performed using Yeasen's one-step RT-qPCR kit (Cat: 11143ES80) according to the instructions. ΔΔCt determination was performed in ABI QuantStudio TM Real-time fluorescence PCR was performed in a 6-well real-time fluorescence PCR system. Each duplex was tested for three independent transfections, and each transfection was performed in triplicate.

[0352] The results of the in vitro activity test of the duplexes are shown in Table 3. As shown in Table 3, duplexes 1-124 all have good inhibitory activity on MMP7 mRNA expression.

[0353] Table 3: Results of single-dose duplex testing in A549 cells

[0354] 2-4. IC of duplexes on A549 and HCC-78 cells 50 and KDmax detection

[0355] A549 and HCC-78 cells (Beina Biotechnology) were cultured and transfected under the same conditions as in 2-1. 50 During the assay, siRNA was tested at duplex concentrations of 10 nM, 0.1 nM, 0.01 nM, 0.0025 nM, 0.001 nM, 0.0004 nM, 0.0001 nM, and 0.000001 nM. RNA was extracted 24 hours after transfection using the same method as in 2-2. The inhibitory activity of the duplex at each concentration point was determined by real-time fluorescence quantitative PCR using the same method as in 2-3. Dose-effect curves were plotted using GraphPad Prism software and the IC was calculated. 50 The highest knockdown rate obtained at the above concentration point is the KDmax value of the duplex.

[0356] Duplex IC in A549 and HCC-78 cells 50 The results of the KDmax tests are shown in Tables 4 and 5, respectively. As shown in Table 4, the IC values ​​of the 13 duplexes tested in A549 cells were 50 All of them were less than 10 pM, and their KDmax were all above 90%. At the same time, as shown in Table 5, the IC 50 The KDmax was also above 95%.

[0357] Table 4: Duplex IC in A549 cells 50 and KDmax test results

[0358] Table 5: Duplex IC in HCC-78 cells 50 and KDmax test results

[0359] 2-5. Activity Detection of Chemically Modified Duplexes in A549 and HCC-78 Cells

[0360] The activity detection method of the chemically modified duplex on A549 cells is the same as 2-1 to 2-3.

[0361] The results of in vitro activity tests of the modified duplexes are shown in Table 6. As shown in Table 6, the in vitro inhibitory activities of duplexes 125-172, 177, 180-185, and 187-237 on MMP7 mRNA expression at 0.1 nM can reach over 30%.

[0362] Table 6: Results of single-dose testing of modified duplexes in A549 cells

[0363] 2-6. IC of chemically modified duplexes in A549 and HCC-78 cells 50 and KDmax detection

[0364] The IC50 and KDmax detection methods of the chemically modified duplexes on A549 and HCC-78 cells are the same as those in 2-4.

[0365] Modified duplex IC in A549 and HCC-78 cells 50 The results of the KDmax tests are shown in Tables 7 and 8, respectively. As shown in Table 7, the IC values ​​of the modified duplexes in A549 cells were 50 All of them were less than 5 pM, and KDmax were all above 90%. At the same time, as shown in Table 8, the IC values ​​of the 18 modified duplexes on HCC-78 cells were 50 The KDmax was also above 95%.

[0366] Table 7: IC50 and KDmax test results of modified duplexes in A549 cells

[0367] Table 8: IC50 and KDmax test results of modified duplexes in HCC-78 cells

[0368] Example 3: In vitro stability test of chemically modified duplexes

[0369] 3-1. Mouse serum stability test:

[0370] siRNA was added to the stability serum (mouse serum: DMEM = 5:4 (volume ratio)) matrix with a final siRNA concentration of 1 μM and a total system of 50 μL. Three replicates were performed in each group as 48h stability samples. Another group of stability serum without siRNA was taken as the 0h sample. Both groups of samples were incubated at 37°C for 48h. After incubation, 150 μL of Loading Buffer was added to the 48h stability sample to terminate the reaction. 150 μL of Loading Buffer was added to the 0h stability sample before adding siRNA. The amount of siRNA added was the same as that of the 48h sample group. After mixing evenly, 50 μL of 100nM internal standard siRNA was added to all samples, purified using a Clarity OTX SPE column, and finally detected by LC-MS the response of the positive and antisense chains of each group of samples. The residual amount was obtained by comparing the response ratio of the 48h and 0h samples.

[0371] 3-2. Mouse lung homogenate stability test:

[0372] siRNA was added to a 12.5 mg / mL mouse lung homogenate solution for a final siRNA concentration of 1 μM. The total volume was 50 μL, with three replicates per group, to serve as the 72-hour stability sample. A separate group of mouse lung homogenate solution, without siRNA, was collected as the 0-hour sample. Both groups of samples were incubated at 37°C for 72 hours. After incubation, 150 μL of Loading Buffer was added to the 72-hour stability sample to terminate the reaction. 150 μL of Loading Buffer was added to the 0-hour stability sample, followed by the addition of siRNA, using the same amount as the 72-hour sample. After thorough mixing, 50 μL of 100 nM internal standard siRNA was added to all samples. The samples were purified using a Clarity OTX SPE column, and the forward and reverse strand responses of each sample were analyzed by LC-MS. The residual amount was determined by comparing the response ratios of the 72-hour and 0-hour samples.

[0373] 3-3. Lysosome stability test:

[0374] siRNA was added to 0.1 mg / mL pH 5.0 lysosomes (Tritosomes) with a final siRNA concentration of 1 μM. The total volume was 50 μL, with three replicates per group, as the 48-hour stability sample. Another group of lysosomes (Tritosomes) without siRNA was taken as the 0-hour sample. Both groups of samples were incubated at 37°C for 48 hours. After incubation, 150 μL of Loading Buffer was added to the 48-hour stability sample to terminate the reaction. 150 μL of Loading Buffer was added to the 0-hour stability sample, followed by the addition of siRNA. The amount of siRNA added was the same as that for the 48-hour sample group. After mixing thoroughly, 50 μL of 100 nM internal standard siRNA was added to all samples, which were purified using a Clarity OTX SPE column. Finally, the responses of the positive and negative strands of each sample group were detected by LC-MS. The residual amount was determined by comparing the response ratio of the 48-hour and 0-hour samples.

[0375] The stability results of the sense and antisense chains of the modified duplexes in serum, lung homogenate, and lysosomes are shown in Table 9. As shown in Table 9, the sense and antisense chains of the modified duplexes have excellent stability in serum, lung homogenate, and lysosomes.

[0376] Table 9: Test results of serum stability, lung homogenate stability and lysosomal stability of modified duplexes

[0377] Example 4: In vitro fibrosis activity detection experiment

[0378] 4.1 TGF-β1-induced fibrosis in A549 cells

[0379] The A549 cells were cultured under the same conditions as in Example 2. The cells were cultured until close to confluence, and the cells were digested with trypsin and plated. 1×10 5 Cells were cultured overnight in 0.5 mL of serum-free medium. After overnight culture, the medium was replaced with serum-free medium. The sample group was transfected with the modified duplex at a final concentration of 2 nM using lipofectamine 2000 (Invitrogen). The blank control group and the TGF-β1-alone treatment group did not contain the modified duplex. 24 hours later, TGF-β1 was added to the sample group and the TGF-β1-alone treatment group at a final concentration of 10 ng / mL, while the blank control group was added with the corresponding volume of serum-free medium. Culture was continued for another 2 days before RNA extraction and real-time fluorescence quantitative PCR was used to measure MMP7 and collagen1A mRNA levels. RNA extraction and real-time fluorescence quantitative PCR were performed as in Example 2.

[0380] 4.2 Primary type II alveolar epithelial cells

[0381] Primary type II alveolar epithelial cells (purchased from ProBio) were cultured at 37°C in an environment of 5% CO2 using epithelial cell culture medium (purchased from ProBio, supplemented with 10% FBS and 1% double-antibody (penicillin-streptomycin (10,000 U / mL))) until nearly confluent. The cells were then trypsinized and plated, with 6 × 10 cells per well of a 24-well plate. 4 cells and 0.5 mL of culture medium; cell transfection, stimulation and detection methods are the same as in Example 4.1.

[0382] 4.3 Primary lung fibroblasts

[0383] Primary lung fibroblasts (purchased from ProBio) were cultured using fibroblast culture medium (purchased from ProBio, supplemented with 10% FBS and 1% bispecific antibody (penicillin-streptomycin (10,000 U / mL))); other culture conditions, cell transfection, stimulation and detection methods were the same as in Example 4.2.

[0384] The results showed that after transfection of the modified duplexes in Table 10 in the three cells, the knockdown efficiency of MMP7 was greater than 90%. The relative level detection results of collagen 1A mRNA are shown in Table 10. The results show that the modified duplexes can downregulate the expression of collagen 1A mRNA induced by TGF-β1.

[0385] Table 10: Relative expression levels of Collagen 1A mRNA in fibrosis experiments

[0386] Example 5: In vivo activity detection

[0387] The modified duplexes used in the in vivo activity assays were all connected to the αvβ6 integrin ligand as the target head at the 5' end of the sense strand via a linker. The linker and target head used the tridentate αvβ6 epithelial cell targeting ligand Tri-SM6.1-αvβ6-(TA14) shown in Figure 1 of patent WO2023070082A2. The names of the modified duplexes after coupling were named by adding the suffix -TRiM to the original numbering.

[0388] 5.1 In vivo activity detection in MMP7+ / - heterozygous mice

[0389] In vivo administration: Commercial MMP7+ / - heterozygous mice (purchased from Biocytogen) were transgenic mice containing the full-length human MMP7 gene (CDS+UTR). In the in vivo experiment, the 0.5 mg / kg dose group consisted of 4 male mice, 6-8 weeks old; the 3 mg / kg dose group consisted of 4 female mice, 6-8 weeks old. 0.5 mg / kg or 3 mg / kg of candidate siRNA molecules were delivered to the lungs via a nebulizer airway. The saline group was delivered with saline, with a nebulization volume of 50 μl each time. Mice were anesthetized and sacrificed 3 or 7 days after treatment, and lung tissue was obtained after perfusion to detect the expression of MMP7 mRNA levels.

[0390] RNA expression detection: After removing the lung tissue, add tissue protection solution and place it at 4°C overnight, then place it in a -80°C refrigerator for long-term storage. Discard the tissue protection solution, add the prepared tissue lysis solution according to the instructions of TaKaRa Mini BEST Universal RNA Extraction Kit (Takara: 9767), and add magnetic beads to homogenize on a tissue homogenizer. After homogenization, perform the subsequent RNA extraction steps according to the instructions. Use the Takara reverse transcription kit (RR036A) to reverse transcribe the extracted RNA, and use the reverse transcribed cDNA to Green kit (ACR, AG11718) was used to amplify the target gene expression level. GAPDH was used as the internal reference and the ΔΔCt method was used in ABIQuantStudio TM Real-time fluorescence PCR was performed in a 6-well real-time fluorescence PCR system with 2-3 replicate wells for each mouse lung tissue sample.

[0391] The results of the in vivo activity test of the modified duplex are shown in Table 11. The results show that the modified duplex can effectively downregulate MMP7 mRNA expression in vivo.

[0392] Table 11: MMP7 mRNA expression levels in MMP7+ / - heterozygous mice after aerosolized airway administration

[0393] 5.2 In vivo activity detection in MMP7 homozygous mice

[0394] In vivo administration: Commercial MMP7 homozygous mice (purchased from Biocytogen), the transgenic mice contain the full-length human MMP7 gene (CDS+UTR). In the in vivo experiment, 3-4 female mice, 6-8 weeks old, were administered to each group. 3 mg / kg of candidate siRNA molecules were delivered to the lungs by nebulized needle airway administration. The saline group delivered saline with a nebulization volume of 50 μl each time. The mice were anesthetized and killed 15 days after administration. The alveolar lavage fluid was first obtained with pre-cooled PBS, and the supernatant was centrifuged for protein quantification (Pierce BCA Protein Assay Kits (Thermo-23225), and MMP7 protein expression was detected using the MMP7 detection kit (Wuhan Cloud-Clone Technology Co., Ltd.-SEA102Hu). After perfusion, lung tissue was taken to detect the mRNA level expression of MMP7. The methods for mouse lung tissue processing, RNA extraction and RNA expression detection were the same as in Example 5.1.

[0395] The results of the in vivo activity test of the modified duplex are shown in Table 12. The results show that the modified duplex can effectively downregulate the expression of MMP7 mRNA and MMP7 protein in vivo.

[0396] Table 12: MMP7 mRNA and protein expression levels in MMP7 homozygous mice 15 days after aerosolized airway administration

[0397] Although the specific embodiments of the present invention have been described in detail, those skilled in the art will understand that various modifications and changes can be made to the details based on all the teachings published, and these changes are all within the scope of protection of the present invention. The entire invention is given by the appended claims and any equivalents thereof.

Claims

1. A small interfering RNA (siRNA) for inhibiting MMP7 gene expression, the siRNA comprising a sense strand and an antisense strand, wherein: The antisense strand comprises at least 17 consecutive nucleotides that differ from the nucleotide sequence shown in any one of SEQ ID NO: 1 to SEQ ID NO: 124 by no more than 4 (e.g., 0, 1, 2, 3 or 4) nucleotides, and the positive strand is at least partially complementary to the antisense strand.

2. The siRNA according to claim 1, wherein The antisense strand comprises at least 17 consecutive nucleotides that differ from the nucleotide sequence shown in any one of SEQ ID NO: 1 to SEQ ID NO: 124 by 0 or 1 nucleotide.

3. The siRNA according to claim 1 or 2, wherein The sense strand comprises at least 17 consecutive nucleotides that differ from the nucleotide sequence shown in any one of SEQ ID NO: 125 to SEQ ID NO: 248 by no more than 4 (e.g., 0, 1, 2, 3 or 4) nucleotides; Preferably, the sense strand comprises at least 17 consecutive nucleotides that differ by 0 or 1 nucleotide from the nucleotide sequence shown in any one of SEQ ID NO: 125 to SEQ ID NO: 248; Preferably, the sense strand has a region of at least 85% complementarity with the antisense strand within the 17 consecutive nucleotides.

4. The siRNA according to any one of claims 1 to 3, wherein The siRNA comprises a blunt end and / or an overhang of 1 to 4 nucleotides; Preferably, the siRNA comprises an overhang of 1 or 2 nucleotides; Preferably, the overhang is present at the 5' end and / or the 3' end of the antisense strand and / or the sense strand; Preferably, the 3' end of the antisense strand of the siRNA comprises an overhang of 2 nucleotides.

5. The siRNA according to any one of claims 1 to 4, wherein The lengths of the antisense strand and the sense strand are independently 17 to 30 nucleotides; preferably, the length of the antisense strand is 19 to 27 nucleotides; preferably, the length of the sense strand is 17 to 25 nucleotides.

6. The siRNA according to any one of claims 1 to 5, wherein The antisense strand is 21 to 23 nucleotides in length, and the sense strand is 19 to 21 nucleotides in length.

7. The siRNA according to any one of claims 1 to 6, wherein The sense strand has no more than 6 (eg, 0, 1, 2, 3, 4, 5, or 6) nucleotide mismatches with the antisense strand.

8. The siRNA according to any one of claims 1 to 7, wherein The sequences of the sense strand and the antisense strand of the siRNA are selected from the sense strand and the antisense strand sequences of any duplex 1 to duplex 124 described in Table 1.

9. The siRNA according to any one of claims 1 to 8, wherein The siRNA contains at least one modified nucleotide.

10. The siRNA according to claim 9, wherein All nucleotides in the sense strand and / or antisense strand of the siRNA are modified nucleotides or nucleotide analogs.

11. The siRNA according to claim 10, wherein The modified nucleotide or nucleotide analog is selected from 2'-methoxy nucleotides, 2'-fluoro nucleotides, 2'-deoxy nucleotides, 2',3'-split ring nucleotide analogs, 2'-fluoroarabino nucleotides, 2'-methoxyethyl nucleotides, 2'-amino modified nucleotides, 2'-alkyl modified nucleotides, 3'-methoxy nucleotides, 2'-allyl modified nucleotides, nucleotides containing thiophosphate groups, nucleotides containing methylphosphonate groups, nucleotides containing 5'-phosphate esters, nucleotides containing 5'-phosphate mimetics, diol-modified nucleotides, abasic nucleotides, morpholino nucleotides, locked nucleotides, unlocked nucleotides or glycerol nucleotides.

12. The siRNA according to any one of claims 9 to 11, wherein The nucleotides in the sense strand of the siRNA are selected from at least two of 2'-methoxy nucleotides, 2'-fluoro nucleotides, and deoxy nucleotides; and / or the nucleotides in the antisense strand of the siRNA are selected from 2'-methoxy nucleotides and 2'-fluoro nucleotides.

13. The siRNA according to any one of claims 9 to 12, wherein The 5' end and / or 3' end of the sense strand of the siRNA optionally comprises a blocking residue (eg, iab).

14. The siRNA according to any one of claims 9 to 13, wherein The first nucleotide at the 5' end of the antisense strand of the siRNA optionally comprises an (E)-vinylphosphonate modification.

15. The siRNA according to any one of claims 1 to 14, wherein The sense strand and / or antisense strand of the siRNA comprises a modified internucleoside linkage; Preferably, the 5' end and the 3' end of the sense strand each independently comprise 1 or 2 phosphorothioate linkages; and / or the 5' end and the 3' end of the antisense strand each independently comprise 1 or 2 phosphorothioate linkages.

16. The siRNA according to any one of claims 9 to 15, wherein The antisense strand of the siRNA comprises the following modification pattern: 1)AS(5'-3'):mN*fN*mNmNmNfNmNfNfNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 380); 2)AS(5'-3'):mN*fN*mNmNmNmNmNmNfNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 381); 3)AS(5'-3'):mN*fN*mNfNmNmNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 382); 4)AS(5'-3'):mN*fN*mNmNmNfNmNfNfNfNmNmNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 383); 5)AS(5'-3'):mN*fN*mNfNmNfNmNfNmNmNmNmNmNfNmNfNmNfNmNfNmN*mN*mN (SEQ ID NO: 384); 6)AS(5'-3'):mN*fN*mNmNmNfNmNmNmNmNmNmNmNfNmNfNmNfNmNmNmN*mN*mN (SEQ ID NO: 385); 7)AS(5'-3'):mN*fN*mNmNmNmNmNfNfNmNmNmNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 386); 8)AS(5'-3'):mN*fN*mNfNmNfNmNfNmNfNmNmNmNfNmNfNmNfNmNfNmN*mN*mN (SEQ ID NO: 387); 9)AS(5'-3'):(E)-VPmN*fN*mNmNmNmNmNfNfNmNmNmNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 388); 10)AS(5'-3'):(E)-VPmN*fN*mNmNmNfNmNfNfNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 389); 11)AS(5'-3'):(E)-VPmN*fN*mNmNmNmNmNmNfNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 390); or, 12)AS(5'-3'):(E)-VPmN*fN*mNmNmNfNmNfNfNfNmNmNmNfNmNfNmNmNmNm NmN*mN*mN (SEQ ID NO: 391); Among them, mN is a methoxy-modified nucleotide, fN is a fluorinated-modified nucleotide, (E)-VP indicates that the nucleotide adjacent to its right is a (E)-vinylphosphonate-modified nucleotide, and * is a phosphorothioate linkage.

17. The siRNA according to any one of claims 9 to 16, wherein The sense strand of the siRNA comprises the following modification pattern: 1) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmNmNmN (SEQ ID NO: 392); 2) SS(5'-3'):mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmN (SEQ ID NO: 393); 3) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNmNmNmNmNmNmN (SEQ ID NO: 394); 4) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmNmN (SEQ ID NO: 395); 5) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNfNmNmNmNmNmNmNmNmN (SEQ ID NO: 396); 6) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmNmNmNmN (SEQ ID NO: 397); 7) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNfNmNmNmNmN (SEQ ID NO: 398); 8) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNfNmNmNmN*mN (SEQ ID NO: 399); 9)SS(5'-3'):mN*mN*mNmNmNmNfNmNdNfNfNfNmNmNmNmNmNfNmNmNmNmN*mN*mN(SEQ ID NO:400); 10)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNfNmNmNmNmN*mN(SEQ ID NO:401); 11)SS(5'-3'):mN*mN*mNmNmNmNmNmNfNfNfNfNmNmNmNmNmNfNfNmNmNmN*mN(SEQ ID NO:402); 12)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNfNmNmNmNmN*mN(SEQ ID NO:403); 13)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNfNmNmNmNmNmNfNmNmN*mN*mN(SEQ ID NO:404); 14)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNfNmNmNmNmNfNfNmNmNmNmN*mN(SEQ ID NO:405); 15)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNfNfNmNmNmN*mN(SEQ ID NO:406); 16)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:407); 17)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNmNmNmNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:408); 18)SS(5'-3'):mN*mN*mNmNmNmNmNmNmNfNmNfNfNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:409); 19)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:410); 20)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:411); 21)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmNmN*(iab)(SEQ ID NO:412); 22) SS(5'-3'):mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmN*(iab) (SEQ ID NO: 413); 23) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmNmN*(iab) (SEQ ID NO: 414); 24)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmNmNmN*(iab)(SEQ ID NO: 415); or, 25)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNfNmNmNmNmN*(iab) (SEQ ID NO: 416); Among them, mN is a methoxy-modified nucleotide, fN is a fluorinated-modified nucleotide, dN is a deoxynucleotide, * is a phosphorothioate linkage, and (iab) is an inverted abasic residue.

18. The siRNA according to any one of claims 9 to 17, wherein The antisense strand comprises the nucleotide sequence shown in any one of SEQ ID NOs: 249-302.

19. The siRNA according to any one of claims 9 to 18, wherein The positive strand comprises the nucleotide sequence shown in any one of SEQ ID NOs: 303-379.

20. The siRNA according to any one of claims 9 to 19, wherein The sequences of the sense strand and antisense strand of the siRNA are selected from the sense strand and antisense strand sequences of any duplex 125 to duplex 172, duplex 177, duplex 180 to duplex 185, and duplex 187 to duplex 237 described in Table 2.

21. A conjugate, wherein The conjugate comprises the siRNA according to any one of claims 1 to 20 and a pharmaceutically acceptable targeting molecule; Preferably, the targeting molecule has affinity for epithelial cell surface receptors; Preferably, the targeting molecule is an integrin ligand, such as an αvβ6 ligand.

22. A pharmaceutical composition, wherein The pharmaceutical composition comprises the siRNA according to any one of claims 1 to 20 or the conjugate according to claim 21, and a pharmaceutically acceptable carrier and / or excipient.

23. The pharmaceutical composition according to claim 22, wherein The pharmaceutically acceptable carrier is a delivery carrier; preferably, the siRNA is encapsulated by the delivery carrier.

24. Use of the siRNA according to any one of claims 1 to 20, the conjugate according to claim 21 or the pharmaceutical composition according to claim 22 or 23 in the preparation of a medicament for treating and / or preventing a pathological condition or disease associated with MMP7; Preferably, the siRNA, the conjugate or the pharmaceutical composition is used alone or in combination with another pharmaceutically active agent (eg, siRNA targeting a different target sequence in the MMP7 gene or siRNA targeting other targets).

25. The use according to claim 24, wherein The pathological condition or disease associated with MMP7 is an inflammatory disease of the lung, such as pulmonary fibrosis (eg idiopathic pulmonary fibrosis, interstitial lung disease) or chronic obstructive pulmonary disease.

26. The use according to claim 24, wherein The pathological condition or disease associated with MMP7 is a fibrotic disease, such as renal fibrosis or liver fibrosis.

27. A method for preventing and / or treating a pathological condition or disease associated with MMP7 in a subject, the method comprising administering to a subject in need thereof an effective amount of the siRNA of any one of claims 1 to 20, the conjugate of claim 21, or the pharmaceutical composition of claim 22 or 23; Preferably, the siRNA, the conjugate or the pharmaceutical composition is used alone or in combination with another pharmaceutically active agent (eg, siRNA targeting a different target sequence in the MMP7 gene or siRNA targeting other targets).

28. The method according to claim 27, wherein: The pathological condition or disease associated with MMP7 is an inflammatory disease of the lung, such as pulmonary fibrosis (eg idiopathic pulmonary fibrosis, interstitial lung disease) or chronic obstructive pulmonary disease.

29. The use according to claim 27, wherein The pathological condition or disease associated with MMP7 is a fibrotic disease, such as renal fibrosis or liver fibrosis.

30. A small interfering RNA (siRNA), the siRNA comprising a sense strand and an antisense strand, wherein: The antisense strand comprises a modification pattern selected from the following 1) to 12): 1)AS(5'-3'):mN*fN*mNmNmNfNmNfNfNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 380); 2)AS(5'-3'):mN*fN*mNmNmNmNmNmNfNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 381); 3)AS(5'-3'):mN*fN*mNfNmNmNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 382); 4)AS(5'-3'):mN*fN*mNmNmNfNmNfNfNfNmNmNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 383); 5)AS(5'-3'):mN*fN*mNfNmNfNmNfNmNmNmNmNmNfNmNfNmNfNmNfNmN*mN*mN (SEQ ID NO: 384); 6)AS(5'-3'):mN*fN*mNmNmNfNmNmNmNmNmNmNmNfNmNfNmNfNmNmNmN*mN*mN (SEQ ID NO: 385); 7)AS(5'-3'):mN*fN*mNmNmNmNmNfNfNmNmNmNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 386); 8)AS(5'-3'):mN*fN*mNfNmNfNmNfNmNfNmNmNmNfNmNfNmNfNmNfNmN*mN*mN (SEQ ID NO: 387); 9)AS(5'-3'):(E)-VPmN*fN*mNmNmNmNmNfNfNmNmNmNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 388); 10)AS(5'-3'):(E)-VPmN*fN*mNmNmNfNmNfNfNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 389); 11)AS(5'-3'):(E)-VPmN*fN*mNmNmNmNmNmNfNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 390); or, 12)AS(5'-3'):(E)-VPmN*fN*mNmNmNfNmNfNfNfNmNmNmNfNmNfNmNmNmNmNmN*mN*mN (SEQ ID NO: 391); Wherein, mN is a methoxy-modified nucleotide, fN is a fluorinated-modified nucleotide, (E)-VP indicates that the nucleotide adjacent to its right side is a (E)-vinylphosphonate-modified nucleotide, and * is a phosphorothioate linkage; and / or, The sense strand comprises a modification pattern selected from the following 1) to 25): 1) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmNmNmN (SEQ ID NO: 392); 2) SS(5'-3'):mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmN (SEQ ID NO: 393); 3) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNmNmNmNmNmNmN (SEQ ID NO: 394); 4) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmNmN (SEQ ID NO: 395); 5) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNfNmNmNmNmNmNmNmNmN (SEQ ID NO: 396); 6) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmNmNmNmN (SEQ ID NO: 397); 7) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNfNmNmNmNmN (SEQ ID NO: 398); 8)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNfNmNmNmNmN*mN(SEQ ID NO:399); 9)SS(5'-3'):mN*mN*mNmNmNmNfNmNdNfNfNfNmNmNmNmNmNfNmNmNmNmN*mN*mN(SEQ ID NO:400); 10)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNfNmNmNmNmN*mN(SEQ ID NO:401); 11)SS(5'-3'):mN*mN*mNmNmNmNmNmNfNfNfNfNmNmNmNmNmNfNfNmNmNmN*mN(SEQ ID NO:402); 12)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNfNmNmNmNmN*mN(SEQ ID NO:403); 13)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNfNmNmNmNmNmNfNmNmN*mN*mN(SEQ ID NO:404); 14)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNfNmNmNmNmNfNfNmNmNmNmN*mN(SEQ ID NO:405); 15)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNfNfNmNmNmN*mN(SEQ ID NO:406); 16)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:407); 17)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNmNmNmNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:408); 18)SS(5'-3'):mN*mN*mNmNmNmNmNmNmNfNmNfNfNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:409); 19)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:410); 20)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO:411); 21)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmNmNmN*(iab) (SEQ ID NO: 412); 22) SS(5'-3'):mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmN*(iab) (SEQ ID NO: 413); 23)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmN mN*(iab) (SEQ ID NO: 414); 24)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmNmN mN*(iab)(SEQ ID NO: 415); 25)SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNfNmNmNmNm N*(iab) (SEQ ID NO: 416); Among them, mN is a methoxy-modified nucleotide, fN is a fluorinated-modified nucleotide, dN is a deoxynucleotide, * is a phosphorothioate linkage, and (iab) is an inverted abasic residue.