siRNA and its compositions

Abstract

This invention provides an siRNA that inhibits the expression of the MMP7 gene, comprising a sense strand and an antisense strand. The siRNA according to this invention has good stability, excellent MMP7 gene inhibitory activity, negligible cytotoxicity and immunostimulant activity, and can significantly reduce MMP7 protein levels at the animal level. It has significant clinical value in the treatment of diseases or conditions that benefit from the reduction of MMP7 levels or inhibition of its expression.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to Chinese Patent Application No. 202310944130.X filed on 28 July 2023 and Chinese Patent Application No. 202410176581.8 filed on 8 February 2024, the entirety of said patent application is incorporated herein by reference. This invention provides siRNA and pharmaceutical compositions thereof that inhibit the expression of matrix metalloproteinase 7 (MMP7). The siRNA and pharmaceutical compositions thereof according to the present invention can treat diseases related to matrix metalloproteinase 7. [Background technology]

[0002] Matrix metalloproteinase 7 (MMP7) is the smallest member of the matrix metalloproteinase (MMP) family, which comprises 24 related secreted zinc-dependent endopeptidases with different 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 of the lung and liver. MMP7 enzyme levels are associated with pathogenic fibrosis through multiple potential mechanisms, including epithelial-mesenchymal transition, extracellular matrix degradation, abnormal matrix repair, and promotion of tissue remodeling. MMP7 activates epithelial cells by cleaving E-cadherin and proteolytically activating heparin-binding epidermal growth factor precursors, thereby releasing active HB-EGF, which promotes epithelial-mesenchymal transition and pulmonary fibroblast proliferation in epithelial cells, ultimately leading to or promoting fibrosis.

[0003] Idiopathic pulmonary fibrosis (IPF) is a generally fatal chronic lung disease with a relatively unpredictable clinical course and disease progression rate. Increased MMP7 expression 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 is associated with IPF severity and progression. Currently, there are only two drug options for IPF patients, pirfenidone and nintedanib, and the median survival time for patients treated with these drugs remains short (approximately 3 years), suggesting the need for more diverse and effective drug options.

[0004] Small interfering RNAs (siRNAs) are novel and highly promising candidate research subjects that can achieve the goal of treating diseases by inhibiting or blocking the expression of target genes of interest in a sequence-specific manner based on the RNA interference mechanism. Specifically, siRNAs are incorporated into RNA-induced silencing complexes (RISCs), and their guide strand, also called the antisense strand, complementaryally pairs with the target nucleic acid in the target gene mRNA, thereby degrading the target gene mRNA and inhibiting or blocking the expression of the target gene. By recognizing the target RNA in a sequence-specific manner through complementary base pairing, siRNAs achieve the goal of cleaving the target RNA, downregulating the target RNA level, and inhibiting the expression of the target gene. Therefore, siRNAs have the potential for a very wide range of applications.

[0005] However, considering the interspecies differences in MMP7, the difficulties in developing siRNA drugs increase. On the other hand, compared to conventional drugs, siRNA has the disadvantage of being less stable and easily degraded by nucleases when administered systemically. Furthermore, there is a need to further enhance its activity while avoiding the induction of side effects such as off-target effects, immune stimulation, and cytotoxicity. Therefore, there is an urgent need to develop candidate siRNAs that are stable in the blood, have good biological activity, and have low cytotoxicity in inhibiting MMP7 gene expression. Meanwhile, developing drugs that can effectively prevent and / or treat MMP7-related diseases using the above-mentioned candidate siRNAs that inhibit MMP7 gene expression requires clinical research and has the potential for commercialization. [Overview of the project]

[0006] This invention provides an siRNA that can effectively inhibit the expression of the MMP7 gene, thereby providing a drug and method for preventing and / or treating MMP7-related diseases.

[0007] siRNA In one embodiment, the present invention provides a small interfering RNA (siRNA) for inhibiting the expression of the MMP7 gene, wherein the siRNA comprises 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 NOs. 1 to SEQ ID NOs. 124 by four or fewer nucleotides (e.g., 0, 1, 2, 3, or 4), and the sense strand and the antisense strand are at least partially complementary.

[0008] The phrase "at least partially complementary" means that the two sequences may be perfectly complementary, or there may be six, five, four, three, two, or one or fewer mismatched base pair formations in total, and together they retain the ability to hybridize under the relevant conditions. Those skilled in the art can determine the conditions best suited for testing the complementarity of the two sequences, depending on the final use of the hybridized nucleotides. Such conditions may be stringent conditions, for example, treatment with 400 mM NaCl, 40 mM PIPES, pH 6.4, 1 mM EDTA, at 50°C or 70°C for 12–16 hours, followed by washing. Other conditions, such as physiologically relevant conditions that may be encountered in living organisms, may also be applied.

[0009] In some embodiments, the antisense strand comprises at least 17 consecutive nucleotides that differ by only 0 or 1 nucleotide from the nucleotide sequence shown in any one of SEQ ID NOs: 1 to SEQ ID NOs: 124.

[0010] In some embodiments, the antisense strand includes at least 17 consecutive nucleotides that differ from the nucleotide sequence shown in any one of SEQ ID NOs. 1 to 124 by four or fewer nucleotides (e.g., 0, 1, 2, 3, or 4). In some embodiments, the antisense strand includes at least 17 consecutive nucleotides that differ from the nucleotide sequence shown in any one of SEQ ID NOs. 1 to 124 by three or fewer nucleotides (e.g., 0, 1, 2, or 3). In some embodiments, the antisense strand includes at least 17 consecutive nucleotides that differ from the nucleotide sequence shown in any one of SEQ ID NOs. 1 to 124 by two or fewer nucleotides (e.g., 0, 1, or 2). In some embodiments, the antisense strand includes at least 17 consecutive nucleotides that differ from the nucleotide sequence shown in any one of SEQ ID NOs. 1 to 124 by one or fewer nucleotides (e.g., 0 or 1). In some embodiments, the antisense strand includes at least 17 consecutive nucleotides of the nucleotide sequence shown in any one of SEQ ID NOs. 1 to 124. In some embodiments, the antisense strand is a nucleotide sequence shown in any one of SEQ ID NOs: 1 to 124.

[0011] In some embodiments, the sense strand and the antisense strand have a mismatch of 6 or fewer nucleotides. In some embodiments, the sense strand and the antisense strand have a mismatch of 5 or fewer nucleotides. In some embodiments, the sense strand and the antisense strand have a mismatch of 4 or fewer nucleotides. In some embodiments, the sense strand and the antisense strand have a mismatch of 3 or fewer nucleotides. In some embodiments, the sense strand and the antisense strand have a mismatch of 2 or fewer nucleotides. In some embodiments, the sense strand and the antisense strand have a mismatch of 1 or fewer nucleotides. In some embodiments, the sense strand and the antisense strand are perfectly complementary.

[0012] In some embodiments, the sense strand and the antisense strand are complementary by at least 15, 16, 17, 18, 19, 20, or 21 nucleotides.

[0013] In some embodiments, the sense strand and the antisense strand have at least 85% (e.g., at least 90%, at least 95%, at least 99%) complementarity or complete complementarity in at least 17 consecutive nucleotides.

[0014] In some embodiments, the sense strand and the antisense strand form double-stranded regions having a length of 15 to 30 nucleotide pairs, for example, 15 to 25 nucleotide pairs, 15 to 23 nucleotide pairs, 15 to 21 nucleotide pairs, for example, 15, 16, 17, 18, 19, 20, or 21 nucleotide pairs.

[0015] In some embodiments, the length of the antisense chain is 17-30 nucleotides (e.g., 17-29, 17-28, 17-27, 19-30, 19-29, 19-28, 19-27, 19-25, 19-23, 21-25, 21-23), and the length of the sense chain is 17-30 nucleotides (e.g., 17-29, 17-28, 17-27, 17-26, 17-25, 17-21, 19-21).

[0016] In some embodiments, the length of the antisense strand is 19 to 27 nucleotides, and the length of the sense strand is 17 to 25 nucleotides.

[0017] In some embodiments, the length of the antisense strand is 21 to 23 nucleotides, and the length of the sense strand is 19 to 21 nucleotides.

[0018] In some embodiments, the length of the antisense strand is 23 nucleotides, and the length of the sense strand is 21 nucleotides.

[0019] In some embodiments, the siRNA includes a blunt end and / or an overhang.

[0020] In some embodiments, the siRNA includes one or more single-stranded nucleotide overhangs, for example, overhangs of 1, 2, 3, or 4 nucleotides. In some embodiments, the overhangs may be located on the sense strand, the antisense strand, or any combination thereof. In some embodiments, the overhangs may be located at the 5' end, 3' end, or both ends of the siRNA antisense strand or sense strand.

[0021] In some embodiments, the 3' end of the antisense strand of the siRNA includes an overhang of two nucleotides. In some embodiments, the 3' end of the sense strand of the siRNA is blunt.

[0022] In some embodiments, the sense strand of the siRNA comprises at least 17 consecutive nucleotides that differ from the nucleotide sequence shown in any one of SEQ ID NOs: 125 to SEQ ID NOs: 248 by 4 or fewer (e.g., 0, 1, 2, 3, or 4) nucleotides. The at least 17 consecutive nucleotides contained in the sense strand and the antisense strand of the siRNA form a double-stranded region. 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 NOs: 125 to SEQ ID NOs: 248 by 3 or fewer (e.g., 0, 1, 2, or 3) nucleotides. 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 NOs: 125 to SEQ ID NOs: 248 by 2 or fewer (e.g., 0, 1, or 2) nucleotides. 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 NOs: 125 to SEQ ID NOs: 248 by 1 or fewer (e.g., 0 or 1) nucleotides. In some embodiments, the sense strand has a region that is at least 85% (e.g., at least 90%, at least 95%, at least 99%) complementary or completely complementary 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 NOs: 125 to SEQ ID NOs: 248. In some embodiments, the sense strand is the nucleotide sequence shown in any one of SEQ ID NOs: 125 to SEQ ID NOs: 248.

[0023] In some embodiments, the antisense strand of the siRNA comprises the antisense strand sequence of any one of duplexes 1 to duplex 124 according to Table 1 or a part 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 part thereof (e.g., at least 17 consecutive nucleotides thereof).

[0024] In some embodiments, the sequences of the sense strand and the antisense strand of the siRNA are selected from the sense strand and antisense strand sequences of any one of duplexes 1 to 124 shown in Table 1.

[0025] 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 according to Table 1 or a part thereof (for example, at least 17 consecutive nucleotides thereof). Preferably, the sense strand of the siRNA comprises the sense strand sequence of the duplex or a part thereof (for example, at least 17 consecutive nucleotides thereof).

[0026] 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 according to Table 1 or a part thereof (for example, at least 17 consecutive nucleotides thereof). Preferably, the sense strand of the siRNA comprises the sense strand sequence of the duplex or a part thereof (for example, at least 17 consecutive nucleotides thereof).

[0027] In some embodiments, the antisense strand of the siRNA includes the antisense strand sequences of double-stranded 24, double-stranded 34, double-stranded 50, double-stranded 53, double-stranded 59, double-stranded 72, double-stranded 75, double-stranded 87, double-stranded 42, double-stranded 94, double-stranded 1, double-stranded 35, double-stranded 79, double-stranded 82, double-stranded 25, double-stranded 39, double-stranded 71, double-stranded 112, double-stranded 124, double-stranded 100, double-stranded 109, double-stranded 113, double-stranded 30, double-stranded 105, double-stranded 115, double-stranded 7, double-stranded 51, double-stranded 15, double-stranded 97, and double-stranded 121 as shown in Table 1, or a portion thereof (for example, at least 17 consecutive nucleotides). Preferably, the sense strand of the siRNA includes the double-stranded sense strand sequence or a portion thereof (for example, at least 17 consecutive nucleotides).

[0028] In some embodiments, the antisense strand of the siRNA includes the antisense strand sequences of double-stranded 69, double-stranded 81, double-stranded 88, double-stranded 96, double-stranded 22, double-stranded 32, double-stranded 47, double-stranded 40, double-stranded 21, double-stranded 29, double-stranded 31, double-stranded 45, double-stranded 63, double-stranded 18, double-stranded 70, double-stranded 114, double-stranded 123, double-stranded 41, double-stranded 90, double-stranded 122, double-stranded 84, and double-stranded 10, or a portion thereof (for example, at least 17 consecutive nucleotides). Preferably, the sense strand of the siRNA includes the sense strand sequence of the double-stranded siRNA, or a portion thereof (for example, at least 17 consecutive nucleotides).

[0029] In some embodiments, the antisense strand of the siRNA comprises the antisense strand sequences of double-stranded 6, double-stranded 60, double-stranded 80, double-stranded 118, double-stranded 16, double-stranded 55, double-stranded 116, double-stranded 19, double-stranded 120, double-stranded 13, double-stranded 74, double-stranded 23, double-stranded 56, and double-stranded 76 as shown in Table 1, or a portion thereof (e.g., at least 17 consecutive nucleotides). Preferably, the sense strand of the siRNA comprises the double-stranded sense strand sequence or a portion thereof (e.g., at least 17 consecutive nucleotides).

[0030] In some embodiments, the antisense strand of the siRNA comprises the antisense strand sequence of double-stranded 2, double-stranded 5, double-stranded 8, double-stranded 9, double-stranded 11, double-stranded 12, double-stranded 33, double-stranded 35, double-stranded 38, double-stranded 46, double-stranded 50, double-stranded 67, or double-stranded 73 as shown in Table 1, or a portion thereof (e.g., at least 17 consecutive nucleotides). Preferably, the sense strand of the siRNA comprises the sense strand sequence of the double-stranded siRNA, or a portion thereof (e.g., at least 17 consecutive nucleotides).

[0031] In some embodiments, the antisense strand of the siRNA comprises the antisense strand sequences of double-stranded 2, double-stranded 9, double-stranded 11, double-stranded 12, double-stranded 38, double-stranded 46, double-stranded 67, or double-stranded 73 as shown in Table 1, or a portion thereof (e.g., at least 17 consecutive nucleotides). Preferably, the sense strand of the siRNA comprises the sense strand sequence of the double-stranded siRNA, or a portion thereof (e.g., at least 17 consecutive nucleotides).

[0032] qualification The siRNAs of the present invention are modified in the nucleoside base structure or the ribose-phosphate backbone structure, thereby reducing off-target effects and / or improving the biological stability of the molecule, or improving the physical stability of the double helix formed between the antisense nucleic acid and the sense nucleic acid. Therefore, siRNA sequences containing any modification are also included within the scope of the present invention. siRNA molecules containing ribose nucleoside analogs or derivatives must retain the ability to form double helixes and enable or mediate the specific degradation of target RNA via the RISC pathway.

[0033] In some embodiments of the present invention, the siRNA contains at least one modified nucleotide. The modifications do not necessarily have to be identical for each of the multiple modified ribose nucleosides in the siRNA.

[0034] 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.

[0035] In some embodiments of the present invention, the siRNA comprises a 2'-modified nucleotide.

[0036] In some embodiments of the present invention, the modified nucleotide is selected from 2'-methoxynucleotide, 2'-fluoronucleotide, 2'-deoxynucleotide, 2',3'-seconucleotide analog, 2'-fluoroarabinonucleotide, 2'-methoxyethyl nucleotide, 2'-amino-modified nucleotide, 2'-alkyl-modified nucleotide, 3'-methoxynucleotide, 2'-allyl-modified nucleotide, nucleotide containing a phosphorothioate group, nucleotide containing a methylphosphonate group, nucleotide containing a 5'-phosphate, nucleotide containing a 5'-phosphate mimetic, diol-modified nucleotide, debasalized nucleotide, morpholinonucleotide, locked nucleotide (LNA), unlocked nucleotide (UNA), or glycerol nucleotide (GNA), but the present invention is not limited thereto.

[0037] In some embodiments of the present invention, the antisense strand of the siRNA has a length of 23 nucleotides. 1) AS(5'-3'): mN*fN*mNmNmNfNmNfNmNfNmNfNmNfNmNfNmNmNmNmN*mN*mN(Array No. 280), 2) AS(5'-3'): mN*fN*mNmNmNmNmNmNfNmNmNfNmNfNmNfNmNfNmNmNmNmN*mN*mN(Array No. 281), 3) AS(5'-3'): mN*fN*mNfNmNmNmNmNmNmNmNfNmNfNmNfNmNmNmNmN*mN*mN(Array No. 282), 4) AS(5'-3'): mN*fN*mNmNmNfNmNfNfNfNmNmNfNmNfNmNfNmNmNmNmN*mN*mN(Array No. 283), 5) AS(5'-3'): mN*fN*mNfNmNfNmNfNmNmNmNmNmNfNmNfNmNfNmN*mN*mN(Array No. 284), 6) AS(5'-3'): mN*fN*mNmNmNfNmNmNmNmNmNmNmNfNmNfNmNfNmNmN*mN*mN(Array No. 285), 7) AS(5'-3'): mN*fN*mNmNmNmNmNfNfNmNmNmNfNmNfNmNfNmNmNmNmN*mN*mN(Array No. 286), 8) AS(5'-3'): mN*fN*mNfNmNfNmNfNmNfNmNfNmNfNmNfNmNfNmN*mN*mN(Array No. 287), 9) AS(5'-3'): (E)-VPmN*fN*mNmNmNmNmNfNfNmNmNmNfNmNfNmNmNmNmN*mN*mN(SEQ ID NO: 288) 10) AS(5'-3'): (E)-VPmN*fN*mNmNfNmNfNmNfNmNfNmNfNmNfNmNfNmNmNmN*mN*mN(SEQ ID NO: 389) 11) AS(5'-3'): (E)-VPmN*fN*mNmNmNmNmNmNfNmNmNfNmNfNmNfNmNmNmNmN*mN*mN(SEQ ID NO: 390),, 12) AS(5'-3'): (E)-VPmN*fN*mNmNfNmNfNfNfNfNmNfNmNfNmNfNmNmNmNmN*mN*mN (Alt. 391) Here, mN is a methoxy-modified nucleotide, fN is a fluoro-modified nucleotide, (E)-VP indicates that the nucleotide adjacent to its right is an (E)-vinylphosphonate-modified nucleotide, and * represents a phosphorothioate bond.

[0038] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides in length and includes the following: 1) The antisense strand has 2'-fluoronucleotides at positions 2, 6, 8, 14, and 16 of its 5' end, and the remaining positions have 2'-methoxynucleotides, or 2) The antisense strand has 2'-fluoronucleotides at positions 2, 6, 14, 16, and 18 of its 5' end, and the remaining positions have 2'-methoxynucleotides, or 3) The antisense strand has 2'-fluoronucleotides at positions 2, 6, 14, 16, and 20 at the 5' end, and the remaining positions have 2'-methoxynucleotides, or 4) The antisense strand has 2'-fluoronucleotides at positions 2, 6, 8, 9, 12, 14, and 16 at the 5' end, and the remaining positions have 2'-methoxynucleotides, or 5) The antisense strand has 2'-fluoronucleotides at positions 2, 9, 12, 14, and 16 at the 5' end, and the remaining positions have 2'-methoxynucleotides, or 6) The antisense strand has 2'-fluoronucleotides at positions 2, 4, 12, 14, and 16 of its 5' end, and the remaining positions have 2'-methoxynucleotides, or 7) The antisense strand has 2'-fluoronucleotides at positions 2, 6, 8, 9, 10, 14, and 16 at the 5' end, and the remaining positions have 2'-methoxynucleotides, or 8) The antisense strand has 2'-fluoronucleotides at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20 at the 5' end, and the remaining positions have 2'-methoxynucleotides, or 9) The antisense strand has 2'-fluoronucleotides at positions 2, 8, 9, 14, and 16 of its 5' end, and the remaining positions are 2'-methoxynucleotides, or 10) The 2, 4, 6, 8, 14, 16, 18, and 20 positions at the 5' end of the antisense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides.

[0039] In some embodiments of the present invention, the sense strand of the siRNA is 21 nucleotides in length. 1) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmNmNmN(Array No. 292), 2) SS(5'-3'): MN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmN(Array No. 292), 3) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNfNmNmNmNmNmNmN(SEQ ID NO: 294) 4) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmNmN(Array No. 295), 5) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNfNmNmNmNmNmNmNmNmN(Array No. 296), 6) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNmNmNmNmNmN(Array No. 297) 7) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmNmNmN(Array No. 298), 8) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNmNmNmN*mN(Array 299), 9) SS(5'-3'): mN*mN*mNmNmNmNfNmNdNfNfNmNmNmNmNfNmNmN*mN(Array 400), 10) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNfNmNmN*mN(SEQ ID NO: 401), 11) SS(5'-3'): mN*mN*mNmNmNmNmNmNfNfNfNmNmNmNmNfNmNmN*mN(SEQ ID NO: 402), 12) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNmNmNmN*mN(Array No. 402), 13) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmN*mN*mN(SEQ ID NO: 404), 14) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNfNmNmNmN*mN(Array 405), 15) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNfNmNfNmNmN*mN(SEQ ID NO: 406), 16) SS(5'-3'): mN*mN*mNmNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmNmN mN*mN (Sequence ID 407), 17) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO: 408), 18) SS(5'-3'): mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN(SEQ ID NO: 409), 19) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO: 410) 20) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO: 411) 21) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmNmN*(iab)(SEQ ID NO: 412), 22) SS(5'-3'): mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmN*(iab)(SEQ ID NO: 412), 23) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmN*(iab)(SEQ ID NO: 414) 24) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmNmN*(iab)(SEQ ID NO: 415), ..., 25) SS(5'-3'): Includes mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmNmNmN*(iab)(sequence code 416), Here, mN is a methoxy-modified nucleotide, fN is a fluoro-modified nucleotide, dN is a deoxyribonucleotide, * is a phosphorothioate bond, and (iab) is an inverted debase residue.

[0040] In some embodiments of the present invention, the sense strand of the siRNA is 21 nucleotides in length and comprises the following: 1) The 7, 9, and 11 positions at the 5' end of the sense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides, or 2) The 7th, 9th, and 10th positions of the sense strand's 5' end are 2'-fluoronucleotides, the 11th position is a deoxynucleotide, and the remaining positions are 2'-methoxynucleotides, or 3) The 7, 9, 10, 11, and 13 positions at the 5' end of the sense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides, or 4) The 7, 9, 10, 11, and 14 positions of the 5' end of the sense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides, or 5) The 7, 9, 10, 11, and 15 positions of the 5' end of the sense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides, or 6) The 7, 9, 10, 11, and 17 positions of the 5' end of the sense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides, or 7) The 7, 9, 10, 11, and 19 positions of the 5' end of the sense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides, or 8) The 9, 11, and 13 positions of the 5' end of the sense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides, or 9) The 7, 9, 10, 11, and 16 positions of the 5' end of the sense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides, or 10) The 7, 9, 11, and 17 positions of the 5' end of the sense strand are 2'-fluoronucleotides, the 10 position is a deoxynucleotide, and the remaining positions are 2'-methoxynucleotides, or 11) The 7th, 9th, and 11th positions of the sense strand's 5' end are 2'-fluoronucleotides, the 10th position is a deoxynucleotide, and the remaining positions are 2'-methoxynucleotides, or 12) The 7, 10, 11, and 17 positions of the sense strand's 5' end are 2'-fluoronucleotides, the 9 position is a deoxynucleotide, and the remaining positions are 2'-methoxynucleotides, or 13) The 7, 9, 10, and 17 positions of the 5' end of the sense strand are 2'-fluoronucleotides, the 11 position is a deoxynucleotide, and the remaining positions are 2'-methoxynucleotides, or 14) The 9, 10, 11, and 17 positions of the 5' end of the sense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides, or 15) The 7, 9, 10, 11, 16, and 17 positions of the 5' end of the sense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides, or 16) The 7, 9, 10, 11, 15, and 17 positions at the 5' end of the sense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides.

[0041] In a baseline embodiment, the siRNA is sequence number 380 and sequence number 392, sequence number 380 and sequence number 393, sequence number 380 and sequence number 394, sequence number 380 and sequence number 395, sequence number 380 and sequence number 396, sequence number 380 and sequence number 397, sequence number 380 and sequence number 398, sequence number 380 and sequence number 399, sequence number 380 and sequence number 400, sequence number 380 and sequence number 401, sequence number 380 and sequence number 402, sequence number 380 and sequence number 403, SEQ ID NOs. 380 and 404, SEQ ID NOs. 380 and 405, SEQ ID NOs. 380 and 406, SEQ ID NOs. 380 and 407, SEQ ID NOs. 380 and 408, SEQ ID NOs. 380 and 409, SEQ ID NOs. 380 and 410, SEQ ID NOs. 380 and 411, SEQ ID NOs. 380 and 412, SEQ ID NOs. 380 and 413, SEQ ID NOs. 380 and 414, SEQ ID NOs. 380 and 415, SEQ ID NOs. 380 and 416, SEQ ID NOs. 381 and 392, SEQ ID NOs. 381 and 393, SEQ ID NOs. 381 and 394, SEQ ID NOs. 381 and 395, SEQ ID NOs. 381 and 396, SEQ ID NOs. 381 and 397, SEQ ID NOs. 381 and 398, SEQ ID NOs. 381 and 399, SEQ ID NOs. 381 and 400, SEQ ID NOs. 381 and 401, SEQ ID NOs. 381 and 402, SEQ ID NOs. 381 and 403, SEQ ID NOs. 381 and SEQ ID NO: 404, SEQ ID NO: 381 and 405, SEQ ID NO: 381 and 406, SEQ ID NO: 381 and 407, SEQ ID NO: 381 and 408, SEQ ID NO: 381 and 409, SEQ ID NO: 381 and 410, SEQ ID NO: 381 and 411, SEQ ID NO: 381 and 412, SEQ ID NO: 381 and 413, SEQ ID NO: 381 and 414, SEQ ID NO: 381 and 415, SEQ ID NO: 381 and 416, Sequence IDs 382 and 392, 382 and 393, 382 and 394, 382 and 395, 382 and 396, 382 and 397, 382 and 398, 382 and 399, 382 and 400, 382 and 401, 382 and 402, 382 and 403, 382 and SEQ ID NO: 404, SEQ ID NO: 382 and 405, SEQ ID NO: 382 and 406, SEQ ID NO: 382 and 407, SEQ ID NO: 382 and 408, SEQ ID NO: 382 and 409, SEQ ID NO: 382 and 410, SEQ ID NO: 382 and 411, SEQ ID NO: 382 and 412, SEQ ID NO: 382 and 413, SEQ ID NO: 382 and 414, SEQ ID NO: 382 and 415, SEQ ID NO: 382 and 416, SEQ ID NOs. 383 and 392, SEQ ID NOs. 383 and 393, SEQ ID NOs. 383 and 394, SEQ ID NOs. 383 and 395, SEQ ID NOs. 383 and 396, SEQ ID NOs. 383 and 397, SEQ ID NOs. 383 and 398, SEQ ID NOs. 383 and 399, SEQ ID NOs. 383 and 400, SEQ ID NOs. 383 and 401, SEQ ID NOs. 383 and 402, SEQ ID NOs. 383 and 403, SEQ ID NOs. 383 and SEQ ID NO: 404, SEQ ID NO: 383 and 405, SEQ ID NO: 383 and 406, SEQ ID NO: 383 and 407, SEQ ID NO: 383 and 408, SEQ ID NO: 383 and 409, SEQ ID NO: 383 and 410, SEQ ID NO: 383 and 411, SEQ ID NO: 383 and 412, SEQ ID NO: 383 and 413, SEQ ID NO: 383 and 414, SEQ ID NO: 383 and 415, SEQ ID NO: 383 and 416, SEQ ID NOs. 384 and 392, SEQ ID NOs. 384 and 393, SEQ ID NOs. 384 and 394, SEQ ID NOs. 384 and 395, SEQ ID NOs. 384 and 396, SEQ ID NOs. 384 and 397, SEQ ID NOs. 384 and 398, SEQ ID NOs. 384 and 399, SEQ ID NOs. 384 and 400, SEQ ID NOs. 384 and 401, SEQ ID NOs. 384 and 402, SEQ ID NOs. 384 and 403, SEQ ID NOs. 384 and SEQ ID NO: 404, SEQ ID NO: 384 and 405, SEQ ID NO: 384 and 406, SEQ ID NO: 384 and 407, SEQ ID NO: 384 and 408, SEQ ID NO: 384 and 409, SEQ ID NO: 384 and 410, SEQ ID NO: 384 and 411, SEQ ID NO: 384 and 412, SEQ ID NO: 384 and 413, SEQ ID NO: 384 and 414, SEQ ID NO: 384 and 415, SEQ ID NO: 384 and 416, SEQ ID NOs. 385 and 392, SEQ ID NOs. 385 and 393, SEQ ID NOs. 385 and 394, SEQ ID NOs. 385 and 395, SEQ ID NOs. 385 and 396, SEQ ID NOs. 385 and 397, SEQ ID NOs. 385 and 398, SEQ ID NOs. 385 and 399, SEQ ID NOs. 385 and 400, SEQ ID NOs. 385 and 401, SEQ ID NOs. 385 and 402, SEQ ID NOs. 385 and 403, SEQ ID NOs. 385 and SEQ ID NO: 404, SEQ ID NO: 385 and 405, SEQ ID NO: 385 and 406, SEQ ID NO: 385 and 407, SEQ ID NO: 385 and 408, SEQ ID NO: 385 and 409, SEQ ID NO: 385 and 410, SEQ ID NO: 385 and 411, SEQ ID NO: 385 and 412, SEQ ID NO: 385 and 413, SEQ ID NO: 385 and 414, SEQ ID NO: 385 and 415, SEQ ID NO: 385 and 416, SEQ ID NOs. 386 and 392, SEQ ID NOs. 386 and 393, SEQ ID NOs. 386 and 394, SEQ ID NOs. 386 and 395, SEQ ID NOs. 386 and 396, SEQ ID NOs. 386 and 397, SEQ ID NOs. 386 and 398, SEQ ID NOs. 386 and 399, SEQ ID NOs. 386 and 400, SEQ ID NOs. 386 and 401, SEQ ID NOs. 386 and 402, SEQ ID NOs. 386 and 403, SEQ ID NOs. 386 and SEQ ID NO: 404, SEQ ID NO: 386 and 405, SEQ ID NO: 386 and 406, SEQ ID NO: 386 and 407, SEQ ID NO: 386 and 408, SEQ ID NO: 386 and 409, SEQ ID NO: 386 and 410, SEQ ID NO: 386 and 411, SEQ ID NO: 386 and 412, SEQ ID NO: 386 and 413, SEQ ID NO: 386 and 414, SEQ ID NO: 386 and 415, SEQ ID NO: 386 and 416, SEQ ID NOs. 387 and 392, SEQ ID NOs. 387 and 393, SEQ ID NOs. 387 and 394, SEQ ID NOs. 387 and 395, SEQ ID NOs. 387 and 396, SEQ ID NOs. 387 and 397, SEQ ID NOs. 387 and 398, SEQ ID NOs. 387 and 399, SEQ ID NOs. 387 and 400, SEQ ID NOs. 387 and 401, SEQ ID NOs. 387 and 402, SEQ ID NOs. 387 and 403, SEQ ID NOs. 387 and SEQ ID NO: 404, SEQ ID NO: 387 and 405, SEQ ID NO: 387 and 406, SEQ ID NO: 387 and 407, SEQ ID NO: 387 and 408, SEQ ID NO: 387 and 409, SEQ ID NO: 387 and 410, SEQ ID NO: 387 and 411, SEQ ID NO: 387 and 412, SEQ ID NO: 387 and 413, SEQ ID NO: 387 and 414, SEQ ID NO: 387 and 415, SEQ ID NO: 387 and 416, SEQ ID NOs. 388 and 392, SEQ ID NOs. 388 and 393, SEQ ID NOs. 388 and 394, SEQ ID NOs. 388 and 395, SEQ ID NOs. 388 and 396, SEQ ID NOs. 388 and 397, SEQ ID NOs. 388 and 398, SEQ ID NOs. 388 and 399, SEQ ID NOs. 388 and 400, SEQ ID NOs. 388 and 401, SEQ ID NOs. 388 and 402, SEQ ID NOs. 388 and 403, SEQ ID NOs. 388 and SEQ ID NO: 404, SEQ ID NO: 388 and 405, SEQ ID NO: 388 and 406, SEQ ID NO: 388 and 407, SEQ ID NO: 388 and 408, SEQ ID NO: 388 and 409, SEQ ID NO: 388 and 410, SEQ ID NO: 388 and 411, SEQ ID NO: 388 and 412, SEQ ID NO: 388 and 413, SEQ ID NO: 388 and 414, SEQ ID NO: 388 and 415, SEQ ID NO: 388 and 416, SEQ ID NOs. 389 and 392, SEQ ID NOs. 389 and 393, SEQ ID NOs. 389 and 394, SEQ ID NOs. 389 and 395, SEQ ID NOs. 389 and 396, SEQ ID NOs. 389 and 397, SEQ ID NOs. 389 and 398, SEQ ID NOs. 389 and 399, SEQ ID NOs. 389 and 400, SEQ ID NOs. 389 and 401, SEQ ID NOs. 389 and 402, SEQ ID NOs. 389 and 403, SEQ ID NOs. 389 and SEQ ID NO: 404, SEQ ID NO: 389 and 405, SEQ ID NO: 389 and 406, SEQ ID NO: 389 and 407, SEQ ID NO: 389 and 408, SEQ ID NO: 389 and 409, SEQ ID NO: 389 and 410, SEQ ID NO: 389 and 411, SEQ ID NO: 389 and 412, SEQ ID NO: 389 and 413, SEQ ID NO: 389 and 414, SEQ ID NO: 389 and 415, SEQ ID NO: 389 and 416, SEQ ID NOs. 390 and 392, 390 and 393, 390 and 394, 390 and 395, 390 and 396, 390 and 397, 390 and 398, 390 and 399, 390 and 400, 390 and 401, 390 and 402, 390 and 403, 390 and SEQ ID NOs: 404, 390 and 405, 390 and 406, 390 and 407, 390 and 408, 390 and 409, 390 and 410, 390 and 411, 390 and 412, 390 and 413, 390 and 414, 390 and 415, 390 and 416, SEQ ID NOs: 391 and 392, 391 and 393, 391 and 394, 391 and 395, 391 and 396, 391 and 397, 391 and 398, 391 and 399, 391 and 400, 391 and 401, 391 and 402, 391 and 403, 391 and 404, 391 and 405, SEQ ID NOs: 391 and 405 The sequence includes a modification pattern selected from 91 and sequence number 406, sequence number 391 and sequence number 407, sequence number 391 and sequence number 408, sequence number 391 and sequence number 409, sequence number 391 and sequence number 410, sequence number 391 and sequence number 411, sequence number 391 and sequence number 412, sequence number 391 and sequence number 413, sequence number 391 and sequence number 414, sequence number 391 and sequence number 415, and sequence number 391 and sequence number 416, where the first sequence number corresponds to the antisense chain and the second sequence number corresponds to the sense chain.

[0042] In a preliminary embodiment, the siRNA is distributed between SEQ ID NOs. 380 and 392, SEQ ID NOs. 380 and 393, SEQ ID NOs. 381 and 393, SEQ ID NOs. 382 and 392, SEQ ID NOs. 383 and 394, SEQ ID NOs. 383 and 395, SEQ ID NOs. 384 and 395, SEQ ID NOs. 385 and 396, SEQ ID NOs. 386 and 397, SEQ ID NOs. 386 and 398, SEQ ID NOs. 386 and 399, SEQ ID NOs. 386 and 400, SEQ ID NOs. 386 and 401, SEQ ID NOs. 386 and 402, SEQ ID NOs. 386 and 403, SEQ ID NOs. 386 and 404, SEQ ID NOs. 386 and 405, SEQ ID NOs. 386 and 406, SEQ ID NOs. 387 The sequence number includes a modification pattern selected from sequence number 395, sequence numbers 387 and 394, sequence numbers 388 and 403, sequence numbers 388 and 399, sequence numbers 388 and 407, sequence numbers 388 and 406, sequence numbers 388 and 415, sequence numbers 388 and 416, sequence numbers 388 and 404, sequence numbers 388 and 402, sequence numbers 388 and 411, sequence numbers 389 and 408, sequence numbers 389 and 412, sequence numbers 390 and 409, sequence numbers 390 and 413, sequence numbers 391 and 410, and sequence numbers 391 and 414, where the first sequence number corresponds to the antisense chain and the second sequence number corresponds to the sense chain.

[0043] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 8, 9, 12, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, and 11 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0044] In some embodiments of the present invention, the antisense strand of the siRNA has a length of 23 nucleotides, where the 2, 6, 8, 9, 12, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA has a length of 21 nucleotides, where the 9, 11, and 13 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0045] In some embodiments of the present invention, the antisense strand of the siRNA has a length of 23 nucleotides, where the 2, 9, 12, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA has a length of 21 nucleotides, where the 9, 11, and 13 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0046] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 4, 12, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, and 11 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0047] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 8, 9, 10, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 15 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0048] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 8, 9, 10, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, and 10 positions at the 5' end of the sense strand are 2'-fluoronucleotides, the 11 position is a deoxynucleotide and the remaining positions are 2'-methoxynucleotides.

[0049] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the positions 2, 4, 6, 8, 14, 16, 18, and 20 at the 5' end of the antisense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides; and the sense strand of the siRNA is 21 nucleotides long, where the positions 7, 9, and 10 at the 5' end of the sense strand are 2'-fluoronucleotides, the position 11 is a deoxynucleotide, and the remaining positions are 2'-methoxynucleotides.

[0050] In some embodiments of the present invention, the antisense strand of the siRNA has a length of 23 nucleotides, where the 2, 6, 14, 16, and 18 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA has a length of 21 nucleotides, where the 7, 9, 10, 11, and 13 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0051] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 8, 9, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 16 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0052] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 8, 9, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 17 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0053] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 8, 9, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 11, and 17 positions at the 5' end of the sense strand are 2'-fluoronucleotides, the 10 position is a deoxynucleotide and the remaining positions are 2'-methoxynucleotides.

[0054] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 8, 9, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 10, 11, and 17 positions at the 5' end of the sense strand are 2'-fluoronucleotides, the 9 position is a deoxynucleotide and the remaining positions are 2'-methoxynucleotides.

[0055] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 8, 9, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, and 17 positions at the 5' end of the sense strand are 2'-fluoronucleotides, the 11 position is a deoxynucleotide and the remaining positions are 2'-methoxynucleotides.

[0056] In some embodiments of the present invention, the antisense strand of the siRNA has a length of 23 nucleotides, where the 2, 8, 9, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA has a length of 21 nucleotides, where the 9, 10, 11, and 17 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0057] In some embodiments of the present invention, the antisense strand of the siRNA has a length of 23 nucleotides, where the 2, 8, 9, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA has a length of 21 nucleotides, where the 7, 9, 10, 11, 16, and 17 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0058] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 8, 9, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, 15, and 17 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0059] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 8, 9, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, and 11 positions at the 5' end of the sense strand are 2'-fluoronucleotides, the 10 position is a deoxynucleotide and the remaining positions are 2'-methoxynucleotides.

[0060] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 4, 6, 8, 10, 14, 16, 18, and 20 positions at the 5' end of the antisense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 15 positions at the 5' end of the sense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides.

[0061] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 4, 6, 8, 10, 14, 16, 18, and 20 positions at the 5' end of the antisense strand are 2'-fluoronucleotides, and the remaining positions are 2'-methoxynucleotides. The sense strand of the siRNA is 21 nucleotides long, where the 7, 9, and 10 positions at the 5' end of the sense strand are 2'-fluoronucleotides, the 11 position is a deoxynucleotide, and the remaining positions are 2'-methoxynucleotides.

[0062] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 8, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, and 11 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0063] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 8, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, and 10 positions at the 5' end of the sense strand are 2'-fluoronucleotides, the 11 position is a deoxynucleotide and the remaining positions are 2'-methoxynucleotides.

[0064] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 8, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 13 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0065] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 8, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 14 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0066] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 8, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 15 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0067] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 8, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 17 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0068] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 8, 14, and 16 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 19 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0069] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 14, 16, and 18 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, and 11 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0070] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 14, 16, and 18 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, and 10 positions at the 5' end of the sense strand are 2'-fluoronucleotides, the 11 position is a deoxynucleotide and the remaining positions are 2'-methoxynucleotides.

[0071] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 14, 16, and 18 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 14 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0072] In some embodiments of the present invention, the antisense strand of the siRNA has a length of 23 nucleotides, where the 2, 6, 14, 16, and 18 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA has a length of 21 nucleotides, where the 7, 9, 10, 11, and 15 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0073] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 14, 16, and 18 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 17 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0074] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 14, 16, and 18 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 19 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0075] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 14, 16, and 20 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, and 11 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0076] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 14, 16, and 20 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, and 10 positions at the 5' end of the sense strand are 2'-fluoronucleotides, the 11 position is a deoxynucleotide and the remaining positions are 2'-methoxynucleotides.

[0077] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 14, 16, and 20 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 13 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0078] In some embodiments of the present invention, the antisense strand of the siRNA has a length of 23 nucleotides, where the 2, 6, 14, 16, and 20 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA has a length of 21 nucleotides, where the 7, 9, 10, 11, and 14 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0079] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 14, 16, and 20 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 15 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0080] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 14, 16, and 20 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 17 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0081] In some embodiments of the present invention, the antisense strand of the siRNA is 23 nucleotides long, where the 2, 6, 14, 16, and 20 positions at the 5' end of the antisense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides, and the sense strand of the siRNA is 21 nucleotides long, where the 7, 9, 10, 11, and 19 positions at the 5' end of the sense strand are 2'-fluoronucleotides and the remaining positions are 2'-methoxynucleotides.

[0082] In some embodiments of the present invention, the siRNA of the present invention comprises a modified nucleoside linkage or a modified backbone. The modified nucleoside linkage or backbone includes, but is not limited to, phosphorothioates, 2'-O methoxyethyl (MOE), 2'-fluoro, alkyl phosphates, diphosphorothioates, alkylphosphorothioates, phosphoramidates, urethanes, carbonates, phosphotryesters, acetamidates, carboxymethyl esters, and combinations thereof.

[0083] 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, the phosphodiester bond is replaced with a thiophosphodiester bond by substituting a non-crosslinked oxygen atom in the phosphodiester bond with one sulfur atom.

[0084] In some embodiments of the present invention, the 5' and 3' ends of the sense chain each independently contain one or two phosphorothioate bonds, and / or the 5' and 3' ends of the antisense chain each independently contain one or two phosphorothioate bonds.

[0085] In some embodiments of the present invention, at least one of the bonds between the nucleotides at positions 1 and 2 of the 5' end of the sense strand, between the nucleotides at positions 2 and 3 of the 5' end of the sense strand, between the nucleotides at positions 1 and 2 of the 3' end of the sense strand, between the nucleotides at positions 2 and 3 of the 3' end of the sense strand, between the nucleotides at positions 1 and 2 of the 3' end of the antisense strand, between the nucleotides at positions 2 and 3 of the 3' end of the antisense strand, between the nucleotides at positions 1 and 2 of the 5' end of the antisense strand, and between the nucleotides at positions 2 and 3 of the 5' end of the antisense strand is a phosphorothioate bond; in some embodiments of the present invention, at least four are phosphorothioate bonds; in some embodiments of the present invention, at least six are phosphorothioate bonds; and in some embodiments of the present invention, all eight are phosphorothioate bonds.

[0086] In some embodiments of the present invention, the nucleotides at positions 1 and 2, and positions 2 and 3 of the sense strand's 5' end are linked by a phosphorothioate.

[0087] In some embodiments of the present invention, the nucleotides at positions 1 and 2, and positions 2 and 3 of the sense strand at the 5' end are linked by a phosphorothioate, and the nucleotides at positions 1 and 2 of the 3' end are linked by a phosphorothioate.

[0088] In some embodiments of the present invention, the nucleotides at positions 1 and 2, and positions 2 and 3 of the sense strand at the 5' end are linked by a phosphorothioate, and the nucleotides at positions 1 and 2, and positions 2 and 3 of the 3' end are linked by a phosphorothioate.

[0089] In some embodiments of the present invention, the nucleotides at positions 1 and 2, and positions 2 and 3 of the 3' end of the antisense chain are linked by a phosphorothioate, and the nucleotides at positions 1 and 2, and positions 2 and 3 of the 5' end are linked by a phosphorothioate.

[0090] In some embodiments of the present invention, the nucleotides at positions 1 and 2 of the 5' end of the sense strand, the nucleotides at positions 2 and 3 of the 5' end of the sense strand, the nucleotides at positions 1 and 2 of the 3' end of the antisense strand, the nucleotides at positions 2 and 3 of the 3' end of the antisense strand, the nucleotides at positions 1 and 2 of the 5' end of the antisense strand, and the nucleotides at positions 2 and 3 of the 5' end of the antisense strand are all linked by phosphorothioates.

[0091] In some embodiments of the present invention, the nucleotides at positions 1 and 2 of the 5' end of the sense strand, positions 2 and 3 of the 5' end of the sense strand, positions 1 and 2 of the 3' end of the sense strand, positions 1 and 2 of the 3' end of the antisense strand, positions 2 and 3 of the 3' end of the antisense strand, positions 1 and 2 of the 5' end of the antisense strand, and positions 2 and 3 of the 5' end of the antisense strand are all linked by phosphorothioates.

[0092] In some embodiments of the present invention, the nucleotides at positions 1 and 2 of the 5' end of the sense strand, positions 2 and 3 of the 5' end of the sense strand, positions 1 and 2 of the 3' end of the sense strand, positions 2 and 3 of the 3' end of the sense strand, positions 1 and 2 of the 3' end of the antisense strand, positions 2 and 3 of the 3' end of the antisense strand, positions 1 and 2 of the 5' end of the antisense strand, and positions 2 and 3 of the 5' end of the antisense strand are all linked by phosphorothioates.

[0093] In some embodiments of the present invention, the sense strand may include one or more capping residues or portions referred to as “capping residues.” A “capping residue” is a non-nucleotide compound or other portion that can be incorporated into one or more ends of the nucleotide sequence of the siRNA. In some embodiments of the present invention, the capping residues are located at the 5' end, the 3' end, or both the 5' and 3' ends of the sense strand.

[0094] In some embodiments of the present invention, an inverted debase 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' and / or 3' ends of the sense strand may contain multiple inverted debase deoxyribose moieties as capping residues.

[0095] In some embodiments of the present invention, one or more inverted debase residues (iabs) are added to the 3' end of the sense strand. In some embodiments of the present invention, one or more inverted debase residues (iabs) are added to the 5' end of the sense strand. In some embodiments of the present invention, one or more inverted debase residues are included at or near one or more ends of the siRNA sense strand.

[0096] The inverted debase residue may be linked via a phosphate ester, phosphorothioate, or other nucleoside bond.

[0097] In some embodiments of the present invention, the first nucleotide at the 5' end of the antisense strand comprises (E)-vinylphosphonate and its analogue modifications, such as (E)-VP modification. In some embodiments of the present invention, the first nucleotide may optionally comprise other modifications, such as 2'-modifications (e.g., 2'-methoxy modification).

[0098] In some embodiments of the present invention, the nucleotides at positions 2-8 of the 5' end of the antisense strand are nucleotides modified with glycerol (GNA) and its analogues.

[0099] In some embodiments, the antisense strand comprises a nucleotide sequence shown in any one of sequence numbers 249 to 302.

[0100] In some embodiments, the sense strand includes a nucleotide sequence shown in any one of sequence numbers 303 to 379.

[0101] In some embodiments, the sense strand and antisense strand sequences of the siRNA are selected from one of the following double-stranded sense strand and antisense strand sequences from double-stranded 125-172, double-stranded 177, double-stranded 180-185, and double-stranded 187-237 as shown in Table 2.

[0102] In some embodiments, the antisense strand includes a 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 includes a 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 sense strand and antisense strand sequences of the siRNA are selected from any one of the double-stranded sense strand and antisense strand sequences of double-stranded 143, double-stranded 144, double-stranded 145, double-stranded 146, double-stranded 147, double-stranded 148, double-stranded 153, double-stranded 157, double-stranded 158, double-stranded 167, double-stranded 168, double-stranded 169, double-stranded 170, double-stranded 171, double-stranded 172, double-stranded 198, double-stranded 200, double-stranded 202, double-stranded 203, double-stranded 208, double-stranded 209, double-stranded 210, double-stranded 211, double-stranded 212, double-stranded 213, double-stranded 214, double-stranded 215, double-stranded 216, and double-stranded 217 shown in Table 2.

[0103] In some embodiments, the antisense strand includes a nucleotide sequence shown in any one of SEQ ID NOs: 256, 264, 285, 286, 287, and 288. In some embodiments, the sense strand includes a 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 sense strand and antisense strand sequences of the siRNA are selected from any one of the double-stranded sense strand and antisense strand sequences of double-stranded 143, double-stranded 144, double-stranded 145, double-stranded 146, double-stranded 148, double-stranded 167, double-stranded 169, double-stranded 171, double-stranded 208, double-stranded 209, double-stranded 210, double-stranded 211, double-stranded 212, double-stranded 213, double-stranded 214, double-stranded 215, double-stranded 216, and double-stranded 217 shown in Table 2.

[0104] In another embodiment, the present invention further provides a small interfering RNA (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*mNmNmNfNmNfNmNfNmNfNmNfNmNfNmNmNmN*mN*mN(Alt. 380), 2) AS(5'-3'): mN*fN*mNmNmNmNmNmNfNmNfNmNfNmNfNmNfNmNmNmN*mN*mN(SEQ ID NO: 381), 3) AS(5'-3'): mN*fN*mNfNmNmNmNmNmNmNmNfNmNfNmNfNmNmNmNmN*mN*mN(Array No. 382), 4) AS(5'-3'): mN*fN*mNmNmNfNmNfNfNfNmNfNmNfNmNfNmNmNmNmN*mN*mN(Array No. 383), 5) AS(5'-3'): mN*fN*mNfNmNfNmNfNmNmNmNmNfNmNfNmNfNmN*mN*mN(Array No. 384), 6) AS(5'-2'): mN*fN*mNmNmNfNmNmNmNmNmNmNfNmNfNmNfNmNmN*mN*mN(Array No. 285), 7) AS(5'-2'): mN*fN*mNmNmNmNmNfNfNmNmNmNfNmNfNmNmNmNmN*mN*mN(Array No. 286), 8) AS(5'-3'):mN*fN*mNfNmNfNmNfNmNfNmNfNmNfNmNfNmNfNmN*mN*mN(Alt. 387), 9) AS(5'-3'): (E)-VPmN*fN*mNmNmNmNmNfNfNmNmNmNfNmNfNmNmNmNmN*mN*mN(SEQ ID NO: 388), 10) AS(5'-3'): (E)-VPmN*fN*mNmNfNmNfNmNfNmNfNmNfNmNfNmNfNmNmNmN*mN*mN(SEQ ID NO: 389), 11) AS(5'-3'):(E)-VPmN*fN*mNmNmNmNmNmNfNmNfNmNfNmNfNmNmNmNmN*mN*mN(SEQ ID NO: 390), or, 12) AS(5'-3'):(E)-VPmN*fN*mNmNfNmNfNfNfNmNfNmNfNmNfNmNmNmN*mN*mN(SEQ ID NO: 391) Here, mN is a methoxy-modified nucleotide, fN is a fluoro-modified nucleotide, (E)-VP indicates that the nucleotide adjacent to its right is an (E)-vinylphosphonate-modified nucleotide, and * represents a phosphorothioate bond. and / or, The aforementioned sense chain includes a modification pattern selected from the following 1) to 25): 1) SS(5'-2'): mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmNmN(SEQ ID NO: 392), 2) SS(5'-3'): mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmN(SEQ ID NO: 393), 3) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNfNmNmNmNmNmNmN(SEQ ID NO: 394), 4) SS(5'-2'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmNmN(SEQ ID NO: 395), 5) SS(5'-2'): mN*mN*mNmNmNmNfNmNfNfNfNmNfNmNmNmNmNmNmNmN(Alt. 396), 6) SS(5'-2'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNmNmNmNmNmN(Altcode 397), 7) SS(5'-2'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmNmNmN(Alt. 398), 8) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmN*mN(Alt. 399), 9) SS(5'-2'): mN*mN*mNmNmNmNfNmNdNfNfNmNmNmNmNfNmNmN*mN(Array No. 400), 10) SS(5'-2'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNfNmNmN*mN(SEQ ID NO: 401), 11) SS(5'-3'): mN*mN*mNmNmNmNmNmNfNfNfNmNmNmNmNfNmNmN*mN(SEQ ID NO: 402), 12) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmN*mN(SEQ ID NO: 403), 13) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmN*mN*mN(SEQ ID NO: 404), 14) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNfNmNmN*mN(SEQ ID NO: 405), 15) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNfNmNfNmNfNmNmN*mN(SEQ ID NO: 406), 16) SS(5'-2'):mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmN*mN(SEQ ID NO: 407), 17) SS(5'-2'): mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO: 408), 18) SS(5'-2'): mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN(SEQ ID NO: 409), 19) SS(5'-3'):mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO: 410), 20) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmN*mN*mN(SEQ ID NO: 411), 21) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmN*(iab)(SEQ ID NO: 412), 22) SS(5'-3'): mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmN*(iab)(SEQ ID NO: 413), 23) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmN*(iab)(SEQ ID NO: 414), 24) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmNmN*(iab)(SEQ ID NO: 415), or, 25) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmNmN*(iab)(SEQ ID NO: 416), Here, mN is a methoxy-modified nucleotide, fN is a fluoro-modified nucleotide, dN is a deoxyribonucleotide, * is a phosphorothioate bond, and (iab) is an inverted debase residue.

[0105] In a preliminary embodiment, the siRNA is distributed between SEQ ID NOs. 380 and 392, SEQ ID NOs. 380 and 393, SEQ ID NOs. 381 and 393, SEQ ID NOs. 382 and 392, SEQ ID NOs. 383 and 394, SEQ ID NOs. 383 and 395, SEQ ID NOs. 384 and 395, SEQ ID NOs. 385 and 396, SEQ ID NOs. 386 and 397, SEQ ID NOs. 386 and 398, SEQ ID NOs. 386 and 399, SEQ ID NOs. 386 and 400, SEQ ID NOs. 386 and 401, SEQ ID NOs. 386 and 402, SEQ ID NOs. 386 and 403, SEQ ID NOs. 386 and 404, SEQ ID NOs. 386 and 405, SEQ ID NOs. 386 and 406, SEQ ID NOs. 387 The sequence number includes a modification pattern selected from sequence number 395, sequence numbers 387 and 394, sequence numbers 388 and 403, sequence numbers 388 and 399, sequence numbers 388 and 407, sequence numbers 388 and 406, sequence numbers 388 and 415, sequence numbers 388 and 416, sequence numbers 388 and 404, sequence numbers 388 and 402, sequence numbers 388 and 411, sequence numbers 389 and 408, sequence numbers 389 and 412, sequence numbers 390 and 409, sequence numbers 390 and 413, sequence numbers 391 and 410, and sequence numbers 391 and 414, where the first sequence number corresponds to the antisense chain and the second sequence number corresponds to the sense chain.

[0106] Delivery The siRNA of the present invention can be delivered or introduced by any method known in the art (e.g., delivered or introduced into cells in vitro, or delivered or introduced into a patient in vivo). For example, to perform in vivo delivery, the siRNA may be injected into a tissue site or administered systemically. In vivo delivery may further be performed via a β-glucan delivery system. In vitro introduction into cells includes methods known in the art, such as electroporation and lipofection.

[0107] In some embodiments, the delivery method includes, but is not limited to, viral delivery (retroviruses, adenoviruses, lentiviruses, baculoviruses, AAVs), liposomes (Lipofectamines, cationic DOTAPs, neutral DOPCs), nanoparticles (cationic polymers, PEIs), bacterial delivery (tkRNAi), chemical modification of siRNA for improved stability (LNAs), lipid nanoparticles (LNPs), neutral liposomes (NLs), 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 applicable to the delivery of nucleic acids or oligonucleotides.

[0108] Conjugate In one embodiment, the present invention provides a conjugate comprising at least one siRNA of the present invention and a pharmaceutically acceptable target molecule. The conjugate of the present invention is obtained by conjugating the siRNA of the present invention with a pharmaceutically acceptable target molecule, the conjugate comprising the pharmaceutically acceptable target molecule and an optional linker. The siRNA may be conjugated noncovalently to the target molecule or covalently to the target molecule.

[0109] The pharmaceutically acceptable target molecules may be target molecules commonly used in the field of siRNA administration, which typically enhance the pharmacokinetic or biodistribution properties of the siRNA ligated to them, improving the cell-specific (or organ-specific) distribution and cell-specific (or organ-specific) uptake of the siRNA. Typical target molecules include, but are not limited to, compounds with affinity for cell surface molecules, cell receptor ligands, haptens, antibodies or antibody fragments, and antibody mimetic compounds. In some embodiments, the target molecule includes, but is not limited to, one or more target molecules or derivatives thereof, such as the integrin family, lipophilic molecules (e.g., cholesterol, bile acids, vitamins (e.g., vitamin E), lipid molecules of different chain lengths, polymers (e.g., polyethylene glycol), polypeptides (e.g., cell membrane permeable peptides, aptamers), antibodies, quantum dots, sugars (e.g., lactose, polylactose, mannose, galactose, N-acetylgalactosamine (GalNAc), folate), or receptor ligands expressed by hepatocytes (e.g., asialoglycoproteins, asialoglycosaccharide residues, lipoproteins (e.g., high-density lipoproteins, low-density lipoproteins, etc.), glucagon, neurotransmitters (e.g., adrenaline), growth factors, transferrin, etc.

[0110] The linker may be a linker commonly used in the field of siRNA administration, and may include, but is not limited to, one or more linkers or derivatives thereof, such as an amide linker moiety, an amino linker moiety, a carbonyl linker moiety, a carbamate linker moiety, a urea linker moiety, an ether linker moiety, a disulfide linker moiety, or a succinyl amino linker moiety.

[0111] In some embodiments, the target molecule may be directly or indirectly linked to the siRNA via a linker / binding group. In some embodiments, the target molecule is linked to the siRNA via an unstable, cleavable, or reversible bond or linker. In some embodiments, the target molecule is linked to at least one end of the sense strand and / or antisense strand of the siRNA. In some embodiments, the target molecule is linked to the 5' and / or 3' end of the sense strand. In some embodiments, the target molecule is linked to the 5' and / or 3' end of the antisense strand.

[0112] In some embodiments, the siRNA of the present invention is delivered to target cells or tissues via a target molecule to which it is ligated. In some embodiments, the target molecule includes a ligand for a cell receptor.

[0113] In some embodiments, the target molecule possesses affinity for epithelial cell surface receptors.

[0114] In some embodiments, the target molecule is an integrin ligand.

[0115] In some embodiments, the target molecule comprises an αvβ6 integrin ligand. Integrin αvβ6 is an epithelial-specific integrin that is highly upregulated in damaged lung epithelial cells. In this specification, “αvβ6 integrin ligand” refers to a molecule having affinity for integrin αvβ6 that can facilitate the targeting and delivery of the siRNA ligated to it to integrin αvβ6-expressing cells (e.g., lung epithelial cells). αvβ6 integrin ligands are known to those skilled in the art; for example, see International Patent Application WO2023070082A2. In some embodiments, two or more (e.g., three) αvβ6 integrin ligands are ligated directly to the siRNA of the present invention or via a linker / ligating group. In some embodiments, the two or more (e.g., three) αvβ6 integrin ligands are ligated to the 5' end of the sense strand.

[0116] Pharmaceutical composition In one embodiment, the present invention provides a pharmaceutical composition comprising at least one siRNA of the present invention.

[0117] In some embodiments of the present invention, the pharmaceutical composition contains at least one of the siRNAs described above.

[0118] In some other embodiments of the present invention, the pharmaceutical composition contains at least two (for example, two, three, four, five, six, seven, eight, nine, ten or more, but not limited to these) siRNAs as active ingredients. Preferably, the at least two siRNAs each target different target sequences in the MMP7 gene, thereby acting simultaneously on different target sequences to produce a synergistic effect. Here, "different target sequences" means that there is no overlap between the target sequences, or that there are fewer than five overlapping consecutive nucleotides between the target sequences (for example, four, three, two, one, or zero overlapping consecutive nucleotides). In this case, the at least two siRNAs can be present in any different proportions. Preferably, the at least two siRNAs can exist in a molar ratio of 1:100 to 100:1 with respect to each other, and more preferably, the at least two siRNAs can exist in a molar ratio of 1:10 to 10:1, 1:5 to 5:1, or 1:2 to 2:1 with respect to each other. In some embodiments of the present invention, the at least two siRNAs exist in the same molar ratio.

[0119] In some 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 proportions, 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, or for example, in the same molar ratio.

[0120] In some embodiments, the pharmaceutical composition contains an effective amount of siRNA. “Effective amount” represents the amount of siRNA that can effectively produce a desired pharmacological, therapeutic, or prophylactic effect. For example, if a given clinical treatment is considered effective when a measurable parameter associated with a disease or disorder is reduced by at least 10%, then the therapeutic effective amount of a drug for treating the disease or disorder is the amount required to achieve at least a 10% reduction in that parameter. For example, a therapeutic effective amount of siRNA targeting MMP7 can reduce MMP7 mRNA levels by at least 10%.

[0121] In some embodiments, in the pharmaceutical composition described in any of the above embodiments, the siRNA can be linked to a target molecule to form a conjugate. Therefore, in some embodiments, the pharmaceutical composition contains the conjugate of the present invention.

[0122] In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or excipient.

[0123] In some embodiments, the pharmaceutically acceptable carrier and / or excipient is a delivery carrier. The delivery carrier is a substance that improves the delivery of nucleic acids or oligonucleotides to cells or tissues. Such a substance may be any delivery carrier known in the art that is applicable to the delivery of nucleic acids or oligonucleotides, and includes, but is not limited to, viruses (retroviruses, adenoviruses, lentiviruses, baculoviruses, AAVs), liposomes (Lipofectamines, cationic DOTAPs, neutral DOPCs), nanoparticles (cationic polymers, PEIs), 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), and the like.

[0124] In some embodiments, the siRNA can be encapsulated by the delivery carrier.

[0125] In some embodiments, the siRNA may be directly or indirectly linked to the delivery carrier via a linker / binding group. In some embodiments, the delivery carrier is linked to the siRNA via an unstable, cleavable, or reversible binding or linker. In some embodiments, the delivery carrier is linked to at least one end of the sense strand and / or antisense strand of the siRNA. In some embodiments, the delivery carrier is linked to the 5' end and / or 3' end of the sense strand. In some embodiments, the delivery carrier is linked to the 5' end and / or 3' end of the antisense strand.

[0126] In some embodiments, the pharmaceutical compositions of the present invention are formulated into dosage forms suitable for the intended route of administration, and can be administered, for example, locally (e.g., by direct injection or implantation), systemically, or subcutaneously, intravenously, intraperitoneally, or parenterally, including intracranial (e.g., intraventricular, intrameningeal, or intrathecal), intramuscular, transdermal, respiratory (aerosol), intranasal, oral, rectal, or topical (including buccal and sublingual) administration.

[0127] In some embodiments, the pharmaceutical composition is administered by inhalation, intranasal administration, intratracheal administration, or oropharyngeal inhalation. A formulation suitable for inhalation administration is prepared by incorporating the required amount of active ingredient into a suitable solvent and subsequently sterile filtration. Formulations used for inhalation administration are typically sterile solutions with physiological pH and low viscosity. Salts may be added to the formulation to balance the tension. In some cases, surfactants or co-solvents may be added to increase the solubility of the active ingredient and improve aerosol properties. In some cases, excipients may be added to control viscosity to ensure the size and distribution of aerosolized droplets.

[0128] In some other embodiments, the pharmaceutical composition may be administered by injection, for example, intravenously, intramuscularly, subcutaneously, intradermally, intra-articularly, intraocularly, intraperitoneally, or topically. Pharmaceutical compositions suitable for injection 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). It is desirable that the pharmaceutical composition maintains stability under manufacturing and storage conditions and prevents contamination by microorganisms such as bacteria and fungi. For example, a sterile injection solution may be prepared by incorporating the required dose of the active ingredient into a suitable solvent, optionally incorporating other desired components (including, but not limited to, pH adjusters, surfactants, adjuvants, ionic strength enhancers, diluents, osmotic pressure-maintaining reagents, absorption-delaying reagents, preservatives, or any combination thereof), followed by filtration and sterilization. Alternatively, the sterile injection solution may be manufactured as a sterile lyophilized powder (e.g., by vacuum drying or lyophilization), thereby facilitating storage and use.

[0129] The siRNA of the present invention may be formulated in the form of a drug unit, thereby facilitating administration. A drug unit refers to a physically distinct unit suitable for use as a single dose to a target of treatment, each unit containing a predetermined amount of active ingredient calculated to produce a desired therapeutic effect when combined with a desired drug carrier.

[0130] In this invention, the optimal target response (e.g., therapeutic or prophylactic response) can be obtained by adjusting the administration scheme. For example, it can be administered as a single dose, multiple doses over a certain period, or the dose can be proportionally reduced or increased depending on the urgency of the treatment situation.

[0131] application Inhibition of MMP7 expression The siRNA of the present invention can be used to inhibit MMP7 expression in vitro and / or in vivo.

[0132] In one embodiment, the present invention provides a method for inhibiting MMP7 expression in cells, the method comprising introducing the siRNA, conjugate, or pharmaceutical composition of the present invention into cells. In some embodiments, the method is carried out 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.

[0133] The term "inhibiting MMP7 expression" refers to at least partial suppression of MMP7 gene expression, which may manifest as a detectable decrease in MMP7 mRNA levels. 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 expressed as a decrease in a parameter functionally related to MMP7 gene expression, such as the amount of protein encoded by the MMP7 gene. In principle, MMP7 gene silencing can be measured by any appropriate assay in any cell expressing MMP7 (constitutive expression or genetically engineered expression). Measurements can be performed at multiple time points before, during, and after siRNA administration to assess the effects of siRNA. MMP7 levels or expression can be measured by evaluating mRNA (e.g., by Northern blot or PCR) or protein (e.g., Western blot or ELISA). For example, the effect of siRNA on MMP7 expression can be measured by measuring the transcription rate of the MMP7 gene (e.g., by RT-PCR).

[0134] In some embodiments, administration of the siRNA of the present invention suppresses the expression of the MMP7 gene by at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%. In some embodiments, administration of the siRNA of the present invention suppresses the expression of the MMP7 gene by at least about 60%, 70%, or 80%. In some embodiments, administration of the siRNA of the present invention suppresses the expression of the MMP7 gene by at least about 85%, 90%, or 95%. In some embodiments, administration of the siRNA of the present invention suppresses the expression of the MMP7 gene by at least about 96%, 97%, 98%, 99%, or 100%.

[0135] In some embodiments, the siRNA, the conjugate, or the pharmaceutical composition may be 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 another target).

[0136] In some embodiments, one siRNA according to the present invention is used. In some other embodiments, at least two (for example, two, three, four, five, six, seven, eight, nine, ten or more, but not limited to these) siRNAs according to the present invention are used, preferably, each of the at least two siRNAs targets a different target sequence in the MMP7 gene. In some other embodiments, at least one siRNA according to the present invention and siRNAs targeting other targets (for example, other genes other than MMP7) are used.

[0137] Treatment of MMP7-related diseases The siRNA of the present invention can be used to treat diseases or conditions that benefit from a reduction in MMP7 levels or inhibition of its expression.

[0138] In one embodiment, the present invention provides a method for preventing and / or treating a condition or disease associated with MMP7 in a subject, the method comprising administering an effective amount of the siRNA, conjugate or pharmaceutical composition of the present invention to a subject in need thereof. The present invention further relates to the use of the siRNA, conjugate or pharmaceutical composition of the present invention in the manufacture of drugs for treating and / or preventing conditions or diseases associated with MMP7.

[0139] In some embodiments, the pathological conditions or diseases associated with MMP7 relate to the overexpression of MMP7. Overexpression of MMP7 refers to MMP7 levels (e.g., MMP7 levels present in the plasma or tissue of the subject, preferably in damaged tissue) being higher than normal MMP7 levels (e.g., the corresponding levels in a healthy control).

[0140] In some embodiments, the pathological conditions or diseases associated with MMP7 benefit from a decrease in MMP7 levels or inhibition of its expression.

[0141] In some embodiments, the pathological condition or disease associated with MMP7 is an inflammatory pneumonia, such as pulmonary fibrosis (e.g., idiopathic pulmonary fibrosis, interstitial lung disease) or chronic obstructive pulmonary disease.

[0142] In some embodiments, the pathological condition or disease associated with MMP7 is a fibrotic disease, such as renal fibrosis or hepatic fibrosis.

[0143] In some embodiments, the pathological condition or disease associated with MMP7 is idiopathic pulmonary fibrosis.

[0144] In some embodiments, for certain diseases, if MMP7 expression is elevated, treatment with the siRNA of the present invention can preferably reduce the level or expression of MMP7 to a level that is considered normal in individuals without such disorders.

[0145] In some embodiments, the subject is a mammal, such as a human.

[0146] In some embodiments, the siRNA, the conjugate, or the pharmaceutical composition may be used alone or in combination with another pharmaceutically active agent (for example, siRNA targeting different target sequences in the MMP7 gene or siRNA targeting other targets), for example, simultaneously or sequentially.

[0147] In some embodiments, one siRNA according to the present invention is used. In some other embodiments, at least two (for example, two, three, four, five, six, seven, eight, nine, ten or more, but not limited to these) siRNAs according to the present invention are used, preferably, each of the at least two siRNAs targets a different target sequence in the MMP7 gene. In some other embodiments, at least one siRNA according to the present invention and siRNAs targeting other targets (for example, other genes other than MMP7) are used.

[0148] In some embodiments, the siRNA of the present invention is administered by inhalation, intranasal administration, intratracheal administration, or oropharyngeal inhalation. In some embodiments, the siRNA of the present invention is administered by inhalation via an inhalation device, such as a metered-dose inhaler, or a nebulizer such as a jet nebulizer or vibrating mesh nebulizer, or a soft mist inhaler, thereby being applied to the treatment of inflammatory pneumonia.

[0149] Definition of Terms In this invention, unless otherwise specified, the scientific and technical terms used herein have meanings that are generally understood by those skilled in the art. Furthermore, to better understand this invention, definitions and interpretations of relevant terms are provided below.

[0150] In this specification, unless otherwise specified, uppercase C, G, U, A, and T represent the base composition of a nucleotide, which may include modified or unmodified nucleotides; lowercase m indicates that the nucleotide adjacent to the right of the label m is a 2'-methoxynucleotide; lowercase f indicates that the nucleotide adjacent to the right of the label f ​​is a 2'-fluoronucleotide; lowercase d indicates that the nucleotide adjacent to the right of the label d is a 2'-deoxynucleotide; the label * indicates that the two nucleotides adjacent to the left and right of the label * are linked by a phosphorothioate; (E)-VP indicates that the nucleotide adjacent to its right is an (E)-vinylphosphonate-modified nucleotide; and iab represents an inverted debase residue.

[0151] In this specification, “modified nucleotide” independently refers to a nucleotide having a modified ribose moiety, a modified nucleoside bond, or a modified base. Therefore, the term “modified nucleotide” encompasses substitution, addition, or removal (e.g., using a functional group or atom) of the nucleoside bond, ribose moiety, or base. Suitable modifications for use in the present invention include all types of modifications disclosed herein or known in the art. “Methoxy-modified nucleotide” refers to a nucleotide in which the hydroxyl group at the 2' position of the ribose group is substituted with methoxy. “Fluoro-modified nucleotide” refers to a nucleotide in which the hydroxyl group at the 2' position of the ribose group is substituted with fluorine. “Nucleotide analog” refers to a group that can substitute for a nucleotide in a nucleic acid but has a different structure from adenine-ribose nucleotide, guanine-ribose nucleotide, cytosine-ribose nucleotide, uracil-ribose nucleotide, or thymine-deoxyribose nucleotide. Examples include isonucleotides, bridged nucleic acid (abbreviated as BNA), or acyclic nucleotides.

[0152] Here, the structure of a fluoromodified nucleotide is as follows: JPEG2026528732000001.jpg3637 The structure of a methoxy-modified nucleotide is as follows: JPEG2026528732000002.jpg3738 The structure of a locust-modified nucleic acid nucleotide is shown below: JPEG2026528732000003.jpg3939 In this specification, the term "siRNA" means an RNA molecule capable of sequence-specifically inducing RNAi phenomena, comprising a sense strand and an antisense strand, and having a partially or completely complementary double-stranded structure. In the siRNA according to the present invention, the length of the complementary double-stranded structure may be 17 to 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, such modified nucleotides not significantly weaken or cause loss of the siRNA's function of inhibiting MMP7 gene expression. Currently, there are many methods available in the art for modifying siRNA, including, for example, skeletal 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).

[0153] The term “antisense strand” includes a region substantially complementary to the target sequence. The term “sense strand,” as used herein, refers to a strand of region substantially complementary to the antisense strand. The term “complementary region” refers to a region in the antisense strand that is substantially complementary to the MMP7 mRNA or a region in the sense strand that is substantially complementary to the antisense strand. If the complementary region is not perfectly complementary to the target sequence or the antisense strand, the mismatch may be located in the interior or terminal regions of the molecule. Typically, the most acceptable mismatches are located in terminal regions, e.g., within 5, 4, 3, 2, or 1 nucleotides at the 5' and / or 3' ends.

[0154] In this specification, 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 specific conditions to form a double-stranded structure. “At least partially complementary” means that the two sequences may be fully complementary, or there may be six, five, four, three, two, or one or fewer mismatched base pair formations in total, and they retain the ability to hybridize under the relevant conditions. Furthermore, if two oligonucleotides are designed to form one or more single-stranded overhangs during hybridization, such overhangs should not be considered mismatches in determining complementarity. In this specification, “complementary” sequences may further include, or be entirely formed from, base pairs formed from non-Watson-Crick base pairs and / or non-naturally modified nucleotides, as long as the above hybridization ability requirements are met. Such non-Watson-Crick base pairs include, but are not limited to, G:U fluctuation base pairing or Hoogstein base pairing. Correspondingly, unless otherwise specified herein, “mismatch” means that the bases at the corresponding positions in an siRNA double-stranded molecule are not paired in a complementary manner.

[0155] Those skilled in the art can determine the most suitable conditions for testing the complementarity of two sequences depending on the final use of the hybridized nucleotides. Such conditions may be stringent conditions, for example, treatment with 400 mM NaCl, 40 mM PIPES, pH 6.4, 1 mM EDTA, at 50°C or 70°C for 12–16 hours, followed by washing. Other conditions, such as physiologically relevant conditions that may be encountered in living organisms, may also be applied.

[0156] In this specification, unless otherwise specified, "difference in nucleotide sequence" means a change in the type of base of a nucleotide at the same or corresponding position 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 changes to U, C, G, or dT, dC, dG, then a difference in nucleotide sequence is considered to exist at that position. It should be explained here that if the nucleotide at the same or corresponding position differs from the original nucleotide sequence only in terms of the presence or absence of modification or the type of modification, then a difference in nucleotide sequence is not considered to exist at that position.

[0157] In this specification, unless otherwise specified, the term “pharmaceutically acceptable carrier and / or excipient” means that carriers, delivery carriers, diluents, excipients and / or salts / esters / hydrates they form, etc., are typically chemically or physically compatible with other components constituting a drug dosage form (e.g., the siRNA of the present invention) and physiologically compatible with the subject. “pharmaceutically acceptable carriers and / or excipients” do not exert a therapeutic effect at the desired dose and are not intended to exert a therapeutic effect. Such components may serve to a) assist in the processing of the drug delivery system in the manufacturing process, b) protect, support or enhance the stability, bioavailability or patient acceptability of the active ingredient, c) assist in product identification, and / or d) enhance any other properties of the active ingredient such as overall safety, efficacy, or delivery during storage and use. For example, the siRNA of the present invention may be encapsulated by a delivery carrier. "Pharmacochemically 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, osmotic pressure-maintaining reagents, absorption-delaying reagents, and preservatives. For example, viruses include, but are not limited to, retroviruses, adenoviruses, lentiviruses, baculoviruses, and AAVs. Liposomes include, but are not limited to, lipofectamines, cationic DOTAP, and neutral DOPC. Nanoparticles include, but are not limited to, cationic polymers and PEI. Bacteria include, but are not limited to, tkRNAi. Polymer 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 reagents, such as parabens, chlorobutanol, phenol, and sorbic acid.Reagents that maintain osmotic pressure include, but are not limited to, sugars, NaCl, and their analogues. Reagents that delay absorption include, but are not limited to, monostearates and gelatin. Diluents include, but are not limited to, water, aqueous buffers (e.g., buffered saline), alcohols, and polyols (e.g., glycerol).

[0158] In this specification, unless otherwise specified, the term "inhibition" means a situation in which the expression of a target gene is downregulated by mRNA degradation of the target gene via siRNA. "Downregulation" means a situation in which the expression level of the target gene is reduced by 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% or 100% compared to the absence of siRNA treatment. Here, a 100% reduction in the expression level of the target gene means that there is no detectable level of target gene expression.

[0159] In this specification, the terms “overhang” or “nucleotide overhang” refer to at least one unpaired nucleotide that protrudes from the siRNA double-stranded structure. For example, an overhang exists if the 3' end of one strand of siRNA extends beyond (or vice versa) the 5' end of the other strand. siRNA may contain an overhang of at least one nucleotide, or the overhang may contain at least 2nt, at least 3nt, at least 4nt, at least 5nt or more. The overhang may contain or consist of a nucleotide / nucleoside analog, the nucleotide / nucleoside analog containing a deoxynucleotide / nucleoside. The overhang may be present on the sense strand, the antisense strand, or any combination thereof. The nucleotide of the overhang may be present on the 5' end, the 3' end, or both ends of the antisense or sense strand of siRNA. Correspondingly, the term “blunt end” means the absence of a nucleotide overhang.

[0160] In this specification, the term “prevention” refers to a method taken to prevent or delay the onset of a disease, disorder, or symptom in a subject, and the term “treatment” refers to a method taken 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, symptom reduction, disease extent reduction, stabilization of the disease state (i.e., no worsening), delay or mitigation of disease progression, improvement or mitigation of the disease state, and symptom relief (whether partial or in whole), whether detectable or undetectable. Furthermore, “treatment” may refer to extending survival time compared to a desired survival time (without treatment).

[0161] In this specification, the term “effective dose” means an amount sufficient to achieve, or at least partially achieve, the desired effect. For example, an effective dose for preventing a disease means an amount sufficient to prevent, inhibit, or delay the onset of the disease, and an effective dose for treating a disease means an amount sufficient to cure or at least partially inhibit the disease and its complications in a patient who has the disease. Determining such an effective dose is entirely within the capabilities of those skilled in the art. For example, an effective dose for therapeutic use depends 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 drug administration, and other treatments administered concurrently.

[0162] Beneficial effects of the invention The siRNA of the present invention can effectively inhibit the expression of the MMP7 gene in vitro and / or in vivo, possesses good stability, negligible cytotoxicity and immunostimulant activity, and can significantly reduce MMP7 protein levels at the animal level. Therefore, the siRNA of the present invention can be used to treat diseases or conditions that benefit from reduced levels or inhibition of MMP7 expression, and has significant clinical value in the treatment of pulmonary inflammation or fibrotic diseases.

[0163] The embodiments of the present invention will be described in detail below, along with examples. However, those skilled in the art will understand that the following examples are merely illustrative and do not limit the scope of the present invention. Based on the detailed description of preferred embodiments below, various objectives and advantageous aspects of the present invention will be feasible to those skilled in the art.

[0164] Sequence information JPEG2026528732000004.jpg126165JPEG2026528732000005.jpg217163JPEG2026528732000006.jpg217163JPEG2026528732000007.j pg217163JPEG2026528732000008.jpg217163JPEG2026528732000009.jpg217163JPEG2026528732000010.jpg217163JPEG2026528732 000011.jpg217163JPEG2026528732000012.jpg217163JPEG2026528732000013.jpg217163JPEG2026528732000014.jpg187163JPEG20 26528732000015.jpg37166JPEG2026528732000016.jpg222164JPEG2026528732000017.jpg222164JPEG2026528732000018.jpg222164 JPEG2026528732000019.jpg221164JPEG2026528732000020.jpg222164JPEG2026528732000021.jpg222164JPEG2026528732000022.j pg222164JPEG2026528732000023.jpg221164JPEG2026528732000024.jpg222164JPEG2026528732000025.jpg222164JPEG20265287320 00026.jpg222164JPEG2026528732000027.jpg222164JPEG2026528732000028.jpg222164JPEG2026528732000029.jpg221164JPEG202 6528732000030.jpg222164JPEG2026528732000031.jpg222164JPEG2026528732000032.jpg222164JPEG2026528732000033.jpg180164 [Modes for carrying out the invention]

[0165] The present invention will now be described in more detail in the following non-limiting embodiments.

[0166] Those skilled in the art will know that the examples illustrate the present invention and are not intended to limit the scope of protection claimed in this application. Unless otherwise specified, the experimental methods in the examples are conventional methods. Unless specific conditions are stated in the examples, the methods are carried out according to conventional conditions or conditions suggested by the manufacturer. Unless the manufacturer is specified, the reagents or equipment used are common products that can be purchased commercially.

[0167] Those skilled in the art will recognize that the siRNA described in the present invention can be obtained by conventional siRNA production methods in the art (e.g., solid-phase synthesis and liquid-phase synthesis), and that commercial custom services are available for both solid-phase and liquid-phase synthesis. It will also be apparent to those skilled in the art that modified nucleotide groups can be introduced into the siRNA described in the present invention by using nucleotide monomers having the corresponding modifications. Methods for producing nucleotide monomers having the corresponding modifications are well known to those skilled in the art, and commercially available monomers are also supplied to the market.

[0168] Example 1: siRNA synthesis An siRNA sequence was designed for the MMP7 gene sequence (see NCBI Reference Sequence: NM_002423.5). For the sense strand and antisense strand of the siRNA sequence of the present invention, and the sense strand and antisense strand of the modified double-stranded siRNA sequence, a deoxynucleoside CPG was used as a solid support. The sense strand was synthesized using the solid support, and the antisense strand was synthesized using a universal CPG.

[0169] Sequence synthesis was performed on a 0.2 μmol scale using a 48-channel synthesizer. The phosphoramidite monomer was used at a concentration of 0.05 M, and the activator was 0.3 M BTT.

[0170] Sequences were cleaved and deprotected in 1.5 ml tubes. The 2-position protecting group was removed using AMA in step 1 and triethylamine hydrofluoric acid in step 2. Sequences containing all modifications at the 2-position required ammonia monolysis. The cleaved and deprotected sequences were precipitated using an acetone:ethanol mixture (volume ratio 80:20) and dissolved in RNase-free water. Each sequence was analyzed by LC-MS to confirm sequence accuracy, quantified by spectrophotometer, and purity determined by HPLC.

[0171] After HPLC purification, lyophilization, and quality control, the salt was precipitated with sodium acetate alcohol, desalted using a 3KD ultrafiltration tube, and after desalting, the sense and antisense strands were identified by spectrophotometer quantification. These strands were then mixed in a 1:1 ratio and annealed to form siRNA double strands. The resulting siRNA double strands are shown in Table 1.

[0172] Example 2: Purification and detection of siRNA activity 2-1. Cell Culture and Transfection A549 cell culture: A549 cells (ATCC No: CCL-185) were cultured in F-12K complete medium (Gibco, supplemented with 10% FBS) at 37°C and 5% CO2 until nearly confluent. The cells were then digested with trypsin, seeded, and cultured in a 96-well plate. 8000 A549 cells and 0.1 mL of F-12K medium (Gibco, supplemented with 10% FBS) were added to each well, and the cells were cultured for 24 hours. After transfecting with siRNA using lipofectamine 2000 (Invitrogen), the cells were cultured again at 37°C and 5% CO2 for another 24 hours, followed by RNA extraction. Single-dose experiments were performed at double-stranded 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 selected for detection).

[0173] 2-2. RNA Extraction Using an RNA extraction kit (yeasen, Cat: 18600ES50), follow the instructions provided with the kit, and finally add 170 μL of RNase-free water to collect the RNA.

[0174] 2-3. Real-time fluorescence quantitative PCR One-step RT-qPCR was performed using the yesen one-step RT-qPCR kit (Cat: 11143ES80) following the instructions. A ΔΔCt assay was performed using ABI QuantStudio. TM Real-time fluorescence PCR was performed using a real-time fluorescence PCR system. Three independent transfection tests were performed on each double-stranded sample, and each transfection was measured in three replicates.

[0175] The results of the in vitro activity tests of the double-stranded proteins are shown in Table 3 below. As can be seen from Table 3, all of the double-stranded proteins 1-124 showed good inhibitory activity against MMP7 mRNA expression.

[0176] JPEG2026528732000034.jpg165155JPEG2026528732000035.jpg221150JPEG2026528732000036.jpg221150JPEG2026528732000037.jpg214150 2-4. Detection of double-stranded IC50 and KDmax in A549 and HCC-78 cells The culture and transfection conditions for A549 and HCC-78 cells (Kita-no Biological Laboratory) are the same as in 2-1. 50For the detection experiment, siRNA was administered at double-stranded 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 extraction was performed 24 hours after transfection, using the same RNA extraction method as in 2-2. Inhibitory activity at each concentration point of the double-stranded siRNA was detected by real-time fluorescence quantitative PCR, using the same method as in 2-3. Dose-effect curves were plotted using GraphPad Prism SoftBank, and IC50 was calculated. 50 The value was calculated. The highest knockdown rate obtained at the above concentration points is the KDmax value of the double hemisphere.

[0177] Double-stranded IC in A549 and HCC-78 cells 50 The results of the KDmax tests are shown in Tables 4 and 5, respectively. As can be seen from Table 4, the 13 double strands detected were IC in A549 cells. 50 All of these values ​​were less than 10 pM, and all of them had a KDmax of 90% or more. In addition, as can be seen from Table 5, the eight double-stranded molecules detected showed IC in HCC-78 cells. 50 All of these values ​​are below 10 pM, and KDmax is above 95%.

[0178] JPEG2026528732000038.jpg99123 JPEG2026528732000039.jpg62130 2-5. Detection of the activity of chemically modified double-stranded proteins in A549 and HCC-78 cells The method for detecting activity in chemically modified double-stranded A549 cells is the same as in sections 2-1 to 2-3.

[0179] The results of detecting the in vitro activity of the modified double-stranded proteins are shown in Table 6. As can be seen from Table 6, double-stranded proteins 125-172, 177, 180-185, and 187-237 can all achieve in vitro inhibitory activity of 30% or more against MMP7 mRNA expression under 0.1 nM conditions.

[0180] JPEG2026528732000040.jpg104153JPEG2026528732000041.jpg221147JPEG2026528732000042.jpg221147JPEG2026528732000043.jpg166147 2-6. Detection of IC50 and KDmax of Chemically Modified Double-Strands in A549 and HCC-78 Cells The method for detecting IC50 and KDmax of chemically modified double-strands in A549 and HCC-78 cells is the same as that in 2-4.

[0181] IC of Double-Strands Modified in A549 and HCC-78 Cells 50 The test results of KDmax are as shown in Table 7 and Table 8 respectively. As can be seen from Table 7, for the 18 detected double-strands after modification, the IC in A549 cells 50 is all less than 5 pM, and the KDmax is all 90% or more. At the same time, as can be seen from Table 8, for the 18 detected double-strands after modification, the IC in HCC-78 cells 50 is also less than 5 pM, and the KDmax is all 95% or more.

[0182] JPEG2026528732000044.jpg130137 JPEG2026528732000045.jpg130145 Example 3: In Vitro Stability Test of Chemically Modified Double-Strands 3-1. Mouse Serum Stability Test: siRNA was added to a stable serum (mouse serum: DMEM = 5:4 (volume ratio)) matrix, resulting in a final siRNA concentration of 1 μM. The total volume was 50 μL, with three duplicates set for each group, and these were designated as 48-h stable samples. Separately, one group of stable serum without added siRNA was taken and designated as the 0-h sample. The two sample groups were incubated at 37°C for 48 hours. After incubation was complete, 150 μL of Loading Buffer was added to the 48-h stable sample to stop the reaction. 150 μL of Loading Buffer was added to the 0-h stable sample, followed by the addition of siRNA, ensuring that the amount of siRNA added matched that of the 48-h sample group. After homogeneous mixing, 50 μL of 100 nM internal standard siRNA was added to all samples, and the samples were purified using a ClarityOTXSPE column. Finally, the sense and antisense strand responses of each sample group were detected by LC-MS, and the remaining amount was obtained by comparing the response ratios of the 48-h and 0-h samples.

[0183] 3-2. Stability test of mouse lung homogenate: siRNA was added to a 12.5 mg / mL mouse lung homogenate solution, resulting in a final siRNA concentration of 1 μM. The total volume was 50 μL, with three duplicates set for each group, and these were designated as 72-h stable samples. Separately, one group of mouse lung homogenate solution without siRNA was taken and designated as the 0-h sample. The two sample groups were incubated at 37°C for 72 hours. After incubation was complete, 150 μL of Loading Buffer was added to the 72-h stable sample to stop the reaction. 150 μL of Loading Buffer was added to the 0-h stable sample, followed by the addition of siRNA, ensuring that the amount of siRNA added matched that of the 72-h sample group. After homogeneous mixing, 50 μL of 100 nM internal standard siRNA was added to all samples, and the samples were purified using a ClarityOTXSPE column. Finally, the sense and antisense strand responses of each sample group were detected by LC-MS, and the residual amount was obtained by comparing the ratio of responses between the 72-h and 0-h samples.

[0184] 3-3. Lysosomal stability test: siRNA was added to lysosomes (tritosomes) at a concentration of 0.1 mg / mL and pH 5.0, resulting in a final siRNA concentration of 1 μM. The total volume of the system was 50 μL, with three duplicates set for each group, and these were designated as 48-h stable samples. Separately, one group of lysosomes (tritosomes) without added siRNA was taken and designated as the 0-h sample. Both sample groups were incubated at 37°C for 48 hours. After incubation was complete, 150 μL of Loading Buffer was added to the 48-h stable sample to stop the reaction. 150 μL of Loading Buffer was added to the 0-h stable sample, followed by the addition of siRNA, ensuring that the amount of siRNA added matched that of the 48-h sample group. After homogeneous mixing, 50 μL of 100 nM internal standard siRNA was added to all samples, and the samples were purified using a ClarityOTXSPE column. Finally, the sense and antisense strand responses of each sample group were detected by LC-MS, and the remaining amount was obtained by comparing the ratio of responses between the 48-h and 0-h samples.

[0185] The stability results of the modified double-stranded sense and antisense strands in serum, lung homogenate, and lysosomes are shown in Table 9. As can be seen from Table 9, the modified double-stranded sense and antisense strands exhibit excellent stability in serum, lung homogenate, and lysosomes.

[0186] JPEG2026528732000046.jpg59165 JPEG2026528732000047.jpg199165 Example 4: In vitro fibrosis activity detection experiment 4.1 TGF-β1-induced A549 cell fibrosis The culture conditions for A549 cells were the same as in Example 2. The cells were cultured until nearly confluent, digested with trypsin, seeded, and placed 1 × 10⁶ cells in each well of a 24-well plate. 5After adding individual cells and 0.5 mL of culture medium and culturing overnight, the medium was replaced with serum-free medium. In the sample group, modified double strands were transfected with lipofectamine 2000 (Invitrogen) at a final concentration of 2 nM. Modified double strands were not present in the blank control group and the TGF-β1 monotherapy group. After 24 hours, TGF-β1 at a final concentration of 10 ng / mL was added to the sample group and the TGF-β1 monotherapy group, and a corresponding volume of serum-free medium was added to the blank group. After culturing for two more days, RNA extraction was performed, and the mRNA levels of MMP7 and collagen1A were detected by real-time fluorescence quantitative PCR. The RNA extraction and real-time fluorescence quantitative PCR methods were the same as in Example 2.

[0187] 4.2 Primary type II alveolar epithelial cells Culture of primary type II alveolar epithelial cells (purchased from Sai Baikang): Culturing in an environment of 37°C and 5% CO2 using epithelial cell medium (purchased from Sai Baikang, supplemented with 10% FBS and 1% biantibody (penicillin-streptomycin (10,000 U / mL))) until nearly confluence, digesting the cells with trypsin, seeding, and placing 6 × 10⁶ cells in each well of a 24-well plate. 4 Add the specified number of cells and 0.5 mL of culture medium. The cell transfection, stimulation, and detection methods are the same as in Example 4.1.

[0188] 4.3 Primary lung fibroblasts Primary lung fibroblasts (purchased from Sai Baikang) were cultured in fibroblast medium (purchased from Sai Baikang, supplemented with 10% FBS and 1% biantibody (penicillin-streptomycin (10,000 U / mL))), with other culture conditions, cell transfection, stimulation, and detection methods being the same as in Example 4.2.

[0189] The results show that in all three cell types, after transfecting with the modified double-stranded mRNA shown in Table 10, the knockdown efficiency of MMP7 exceeded 90%, and the relative level detection results of collagen 1A mRNA are shown in Table 10. As can be seen from the results, the modified double-stranded mRNA can downregulate TGF-β1-induced collagen 1A mRNA expression.

[0190] JPEG2026528732000048.jpg168160 Example 5: In vivo activity detection The modified double-stranded molecules used for in vivo activity detection all have an αvβ6 integrin ligand linked to the 5' end of the sense strand via a linker, with the linker and targeting site using the tridentate αvβ6 epithelial cell targeting ligand Tri-SM6.1-αvβ6-(TA14) shown in Figure 1 of patent WO2023070082A2. The conjugated modified double-stranded molecules were named by adding the suffix -TRiM to the original number.

[0191] 5.1 Detection of in vivo activity in MMP7+ / - heterozygous mice In vivo administration: Commercially available MMP7+ / - heterozygous mice (purchased from Biocytogen) were used, and these transgenic mice contained 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 aged 6-8 weeks each, and the 3 mg / kg dose group consisted of 4 female mice aged 6-8 weeks each. Candidate siRNA molecules at 0.5 mg / kg or 3 mg / kg were delivered to the lungs via nebulizer needle, and saline was delivered to the saline group with a spray volume of 50 μl each time. After administration for 3 or 7 days, the mice were sacrificed under anesthesia, and lung tissue was collected after perfusion and used to detect MMP7 mRNA level expression.

[0192] RNA Expression Detection: After extracting lung tissue, tissue preservation solution was added and left overnight at 4°C, then stored in a -80°C refrigerator for long-term storage. The tissue preservation solution was discarded, and tissue lysate prepared according to the instructions for the TaKaRa Mini BEST Universal RNA Extraction Kit (Takara:9767) was added, magnetic beads were added, and the tissue was homogenized using a tissue homogenizer. After homogenization was complete, each subsequent RNA extraction step was performed according to the instructions. The extracted RNA was reverse transcribed using the Takara Reverse Transcription Kit (RR036A), and the resulting cDNA was converted to SYBR (R) The expression level of the target gene was detected by amplification using the Green kit (AG11718). Using GAPDH as the internal standard, the ΔΔCt assay was performed using ABI QuantStudio. TM Real-time fluorescence PCR was performed using a real-time fluorescence PCR system. Two to three wells were prepared in parallel for each mouse lung tissue sample.

[0193] The results of the in vivo activity test of the modified double-stranded mRNA are shown in Table 11. As can be seen from the results, the modified double-stranded mRNA can effectively downregulate MMP7 mRNA expression in vivo.

[0194] JPEG2026528732000049.jpg179166 5.2 Detection of in vivo activity in MMP7 homozygous mice In vivo administration: Commercially available MMP7 homozygous mice (purchased from Biocytogen) were used, and these transgenic mice contained the full-length human MMP7 gene (CDS+UTR). In the in vivo experiments, each group consisted of 3-4 female mice aged 6-8 weeks. Candidate siRNA molecules at a dose of 3 mg / kg were delivered to the lungs via nebulizer needle administration, and physiological saline was delivered to a saline group with a spray volume of 50 μl each time. After 15 days of administration, the mice were sacrificed under anesthesia, and the bronchoalveolar lavage fluid was collected in pre-cooled PBS, centrifuged, and the supernatant was taken for protein quantification (Pierce BCA Protein Assay Kits (Thermo-23225)). MMP7 protein expression was detected using an MMP7 detection kit (Wuhan Yunkelong Technology Co., Ltd. - SEA102Hu). Lung tissue was collected after perfusion and used for detection of MMP7 mRNA level expression. The methods for processing mouse lung tissue, RNA extraction, and RNA expression detection were the same as in Example 5.1.

[0195] The in vivo activity test results for the modified double-stranded protein are shown in Table 12. As can be seen from the results, the modified double-stranded protein can effectively downregulate the expression of MMP7 mRNA and MMP7 protein in vivo.

[0196] JPEG2026528732000050.jpg84166 While specific embodiments of the present invention have been described in detail, those skilled in the art should understand that various modifications and changes can be made to the details based on all the disclosed teachings, and that any of these changes will fall within the scope of the claims of the present invention. The entire scope of the present invention is given by the appended claims and any equivalents thereof.

Claims

1. A small interfering RNA (siRNA) for inhibiting MMP7 gene expression, wherein the siRNA comprises 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 NOs. 1 to SEQ ID NOs. 124 by four or fewer nucleotides (e.g., 0, 1, 2, 3, or 4), and the sense strand and the antisense strand are at least partially complementary.

2. The siRNA according to claim 1, wherein 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 NOs: 1 to SEQ ID NOs:

124.

3. The sense strand comprises at least 17 consecutive nucleotides that differ from the nucleotide sequence shown in any one of SEQ ID NOs. 125 to 248 by four or fewer nucleotides (for example, 0, 1, 2, 3, or 4). 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 NOs: 125 to 248. Preferably, the sense strand has a region within the 17 consecutive nucleotides that is at least 85% complementary to the antisense strand, according to claim 1 or 2.

4. The siRNA includes a blunt end and / or an overhang of 1 to 4 nucleotides. Preferably, the siRNA includes one or two nucleotide overhangs. Preferably, the overhang is located at the 5' and / or 3' ends of the antisense chain and / or the sense chain. Preferably, the siRNA according to any one of claims 1 to 3, wherein the 3' end of the antisense strand of the siRNA includes an overhang of two 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 each independently 17 to 30 nucleotides, preferably the length of the antisense strand is 19 to 27 nucleotides, and 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 length of the antisense strand is 21 to 23 nucleotides, and the length of the sense strand is 19 to 21 nucleotides.

7. The siRNA according to any one of claims 1 to 6, wherein the sense strand and the antisense strand have six or fewer nucleotide mismatches (for example, 0, 1, 2, 3, 4, 5, or 6).

8. The siRNA according to any one of claims 1 to 7, wherein the sense strand and antisense strand sequences of the siRNA are selected from any one of the double-stranded sense strand and antisense strand sequences from double-stranded 1 to double-stranded 124 listed 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 modified nucleotide or nucleotide analog is selected from 2'-methoxynucleotide, 2'-fluoronucleotide, 2'-deoxynucleotide, 2',3'-seconucleotide analog, 2'-fluoroarabinonucleotide, 2'-methoxyethyl nucleotide, 2'-amino modified nucleotide, 2'-alkyl modified nucleotide, 3'-methoxy nucleotide, 2'-allyl modified nucleotide, nucleotide containing a phosphorothioate group, nucleotide containing a methylphosphonate group, nucleotide containing a 5'-phosphate, nucleotide containing a 5'-phosphate mimetic, diol modified nucleotide, debasalized nucleotide, morpholino nucleotide, locked nucleotide, unlocked nucleotide, or glycerol nucleotide, as described in claim 10.

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'-methoxynucleotides, 2'-fluoronucleotides, and deoxynucleotides, and / or the nucleotides in the antisense strand of the siRNA are selected from 2'-methoxynucleotides and 2'-fluoronucleotides.

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 includes a capping residue (e.g., iab).

14. The siRNA according to any one of claims 9 to 13, wherein the nucleotide at position 1 of the 5' end of the antisense strand of the siRNA optionally comprises an (E)-vinylphosphonate modification.

15. The sense strand and / or antisense strand of the siRNA include modified nucleoside bonds, Preferably, the siRNA according to any one of claims 1 to 14, wherein the 5' and 3' ends of the sense strand each independently contain one or two phosphorothioate bonds, and / or the 5' and 3' ends of the antisense strand each independently contain one or two phosphorothioate bonds.

16. The antisense strand of the siRNA includes the following modification pattern: 1) AS(5'-3'): mN*fN*mNmNmNfNmNfNmNfNmNfNmNfNmNfNmNmNmN*mN*mN(Condition 380) 2) AS(5'-3'): mN*fN*mNmNmNmNmNfNmNfNmNfNmNfNmNfNmNmNmN*mN*mN(Conversion 381 instead) 3) AS(5'-3'): mN*fN*mNfNmNmNmNmNmNmNfNmNfNmNfNmNmNmNmN*mN*mN(Condition 382) 4) AS(5'-3'): mN*fN*mNmNmNfNmNfNfNfNmNmNfNmNmNmNmN*mN*mN(Condition 383) 5) AS(5'-3'): mN*fN*mNfNmNfNmNfNmNfNmNmNfNmNfNmNfNmN*mN*mN(Inversion 384) 6) AS(5'-3'): mN*fN*mNmNmNfNmNmNmNmNmNfNmNfNmNfNmNmN*mN*mN(Inversion 385) 7) AS(5'-3'): mN*fN*mNmNmNmNfNfNfNmNmNmNfNmNfNmNmNmNmN*mN*mN(Inverse 386) 8) AS(5'-3'): mN*fN*mNfNmNfNmNfNmNfNmNfNmNfNmNfNmNfNmN*mN*mN(Sequence ID 387), 9) AS(5'-3'): (E)-VPmN*fN*mNmNmNmNfNfNmNmNmNfNmNfNmNmNmNmN*mN*mN(Inversion Number 388) 10) AS(5'-3'): (E)-VPmN*fN*mNmNfNmNfNmNfNmNfNmNfNmNfNmNmNmN*mN*mN (Sequence ID 389), 11) AS(5'-3'): (E)-VPmN*fN*mNmNmNmNmNfNmNfNmNfNmNfNmNfNmNmNmN*mN*mN (Inverse 390), or, 12) AS(5'-3'): (E)-VPmN*fN*mNmNfNmNfNmNfNfNmNfNmNfNmNmNmN*mN*mN(Sequence ID 391), Herein, mN is a methoxy-modified nucleotide, fN is a fluoro-modified nucleotide, (E)-VP indicates that the nucleotide adjacent to its right is an (E)-vinylphosphonate-modified nucleotide, and * is a phosphorothioate bond, the siRNA according to any one of claims 9 to 15.

17. The sense strand of the aforementioned siRNA includes the following modification pattern: 1) SS(5'-3'): mN*mN*mNmNmNmNf 2) SS(5'-3'): mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmN(☐393) 3) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNmNmNmNmN(Concurrent 394) 4) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmNmNmN(concurrent 395 instead 5) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNm 6) SS(5'-3'): mN*mN*mNmNmNfNmNfNfNfNmNmNmNfNmNmNmNmNmN(Condition 397) 7) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmNmNmN(Conversion 398) 8) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNfNmNmN*mN(Conversion 399) 9) SS(5'-3'): mN*mN*mNmNmNmNfNmNdNfNfNmNmNmNmNfNmNmN*mN(inverse 400) 10) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNfNmNmN*mN(inverse 401 instead) 11) SS(5'-3'): mN*mN*mNmNmNmNmNmNfNfNfNmNmNmNmNfNmNmN*mN(inverse 402) 12) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmN*mN(inversion number 403) 13) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNmNmN*mN*mN(Inversion No. 404), 14) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNfNmNmNmN*mN(Inversion 405) 15) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNmNmNfNmNfNmNmN*mN(Inverse 406) 16) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmN*mN(inverse 407 instead) 17) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmN*mN*mN(inversion 408) 18) SS(5'-3'): mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN(Inversion 409), 19) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmN*mN*mN(inverse 410) 20) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmN*mN*mN(inversion number 411 instead 21) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmN*(iab)(Inverse 412) 22) SS(5'-3'): mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmN*(iab)(inverse number 413), 23) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmN*(iab)(Inversion number 414), 24) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmNmN*(iab) (Indicate No. 415), or, 25) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmNmN*(iab)(Sequence ID 416), The siRNA according to any one of claims 9 to 16, wherein mN is a methoxy-modified nucleotide, fN is a fluoro-modified nucleotide, dN is a deoxynucleotide, * is a phosphorothioate bond, and (iab) is an inverted abasic residue.

18. The siRNA according to any one of claims 9 to 17, wherein the antisense strand comprises a nucleotide sequence shown in any one of sequence numbers 249 to 302.

19. The siRNA according to any one of claims 9 to 18, wherein the sense strand comprises a nucleotide sequence shown in any one of sequence numbers 303 to 379.

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

21. A conjugate comprising the siRNA and a pharmaceutically acceptable target molecule as described in any one of claims 1 to 20, Preferably, the target molecule has affinity for epithelial cell surface receptors, Preferably, the target molecule is a conjugate of an integrin ligand, such as an αvβ6 ligand.

22. A pharmaceutical composition comprising an siRNA according to any one of claims 1 to 20 or a 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, and 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 manufacture of a drug for treating and / or preventing a pathological condition or disease related to MMP7, Preferably, the siRNA, the conjugate, or the pharmaceutical composition is used alone or in combination with additional pharmaceutically active agents (e.g., siRNA targeting different target sequences 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 pneumonia, for example, pulmonary fibrosis (e.g., 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 the MMP7 is a fibrotic disease, such as renal fibrosis or hepatic fibrosis.

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

28. The method according to claim 27, wherein the pathological condition or disease associated with MMP7 is an inflammatory pneumonia, such as pulmonary fibrosis (e.g., 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 the MMP7 is a fibrotic disease, such as renal fibrosis or hepatic fibrosis.

30. A small interfering RNA (siRNA), wherein the siRNA comprises a sense strand and an antisense strand, and the antisense strand comprises a modification pattern selected from the following 1) to 12): 1) AS(5'-3'): mN*fN*mNmNmNfNmNfNmNfNmNfNmNfNmNfNmNmNmN*mN*mN(Condition 380) 2) AS(5'-3'): mN*fN*mNmNmNmNmNfNmNfNmNfNmNfNmNfNmNmNmN*mN*mN(Conversion 381 instead) 3) AS(5'-3'): mN*fN*mNfNmNmNmNmNmNmNfNmNfNmNfNmNmNmNmN*mN*mN(Condition 382) 4) AS(5'-3'): mN*fN*mNmNmNfNmNfNfNfNmNmNfNmNmNmNmN*mN*mN(Condition 383) 5) AS(5'-3'): mN*fN*mNfNmNfNmNfNmNfNmNmNfNmNfNmNfNmN*mN*mN(Inversion 384) 6) AS(5'-3'): mN*fN*mNmNmNfNmNmNmNmNmNfNmNfNmNfNmNmN*mN*mN(Inversion 385) 7) AS(5'-3'): mN*fN*mNmNmNmNfNfNfNmNmNmNfNmNfNmNmNmNmN*mN*mN(Inverse 386) 8) AS(5'-3'): mN*fN*mNfNmNfNmNfNmNfNmNfNmNfNmNfNmNfNmN*mN*mN(Sequence ID 387), 9) AS(5'-3'): (E)-VPmN*fN*mNmNmNmNfNfNmNmNmNfNmNfNmNmNmNmN*mN*mN(Inversion Number 388) 10) AS(5'-3'): (E)-VPmN*fN*mNmNfNmNfNmNfNmNfNmNfNmNfNmNmNmN*mN*mN (Sequence ID 389), 11) AS(5'-3'): (E)-VPmN*fN*mNmNmNmNmNfNmNfNmNfNmNfNmNfNmNmNmN*mN*mN (Inverse 390), or, 12) AS(5'-3'): (E)-VPmN*fN*mNmNfNmNfNmNfNfNmNfNmNfNmNmNmN*mN*mN(Sequence ID 391), Here, mN is a methoxy-modified nucleotide, fN is a fluoro-modified nucleotide, (E)-VP indicates that the nucleotide adjacent to its right is an (E)-vinylphosphonate-modified nucleotide, and * represents a phosphorothioate bond. and / or, The sense chain includes a modification pattern selected from the following 1) to 25): 1) SS(5'-3'): mN*mN*mNmNmNmNf 2) SS(5'-3'): mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmN(☐393) 3) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNmNmNmNmNmN(Concurrent 394) 4) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmNmNmN(concurrent 395 instead 5) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNm 6) SS(5'-3'): mN*mN*mNmNmNfNmNfNfNfNmNmNmNfNmNmNmNmNmN(Condition 397) 7) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmNmNmN(Conversion 398) 8) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNfNmNmN*mN(Conversion 399) 9) SS(5'-3'): mN*mN*mNmNmNmNfNmNdNfNfNmNmNmNmNfNmNmN*mN(inverse 400) 10) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNfNmNmN*mN(inverse 401 instead) 11) SS(5'-3'): mN*mN*mNmNmNmNmNmNfNfNfNmNmNmNmNfNmNmN*mN(inverse 402) 12) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmN*mN(inversion number 403) 13) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNmNmN*mN*mN(Inversion No. 404), 14) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNfNfNmNmNmN*mN(Inversion 405) 15) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNmNmNfNmNfNmNmN*mN(Inverse 406) 16) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmN*mN(inverse 407 instead) 17) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmN*mN*mN(inversion 408) 18) SS(5'-3'): mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmN*mN*mN(Inversion 409), 19) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmN*mN*mN(inverse 410) 20) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmN*mN*mN(inversion number 411 instead 21) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNmNfNmNmNmNmNmNmNmNmN*(iab)(Inverse 412) 22) SS(5'-3'): mN*mN*mNmNmNmNmNmNfNmNfNmNfNmNmNmNmNmNmN*(iab)(inverse number 413), 23) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNdNmNmNmNmNmNmNmNmNmN*(iab)(Inversion number 414), 24) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNdNfNmNmNmNmNmNmNmNmNmN*(iab) (Indicate No. 415), or, 25) SS(5'-3'): mN*mN*mNmNmNmNfNmNfNfNfNmNmNmNmNfNmNmNmN*(iab)(Sequence ID 416), Here, mN is a methoxy-modified nucleotide, fN is a fluoro-modified nucleotide, dN is a deoxyribonucleotide, * is a phosphorothioate bond, and (iab) is an inverted abasic residue, representing a small interfering RNA (siRNA).

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