SiRNA for inhibiting PLN gene expression and conjugate and application thereof

Abstract

The invention belongs to the field of biological medicine, and relates to siRNA for inhibiting PLN gene expression and a conjugate, the siRNA comprises a positive-sense strand and an antisense strand, the positive-sense strand comprises a nucleotide sequence I, and the antisense strand comprises a nucleotide sequence II; each nucleotide in the nucleotide sequence I and the nucleotide sequence II is modified or unmodified nucleotide; the nucleotide sequence I and the nucleotide sequence II are at least partially reversely complementary to form a double-stranded region; the nucleotide sequence I is basically consistent with a first section of nucleotide sequence, and the first section of nucleotide sequence is a section of nucleotide sequence with the length of at least 19 nucleotides in mRNA expressed by the PLN gene. The siRNA and the conjugate and the pharmaceutical composition thereof disclosed by the invention can effectively prevent and / or treat diseases associated with PLN expression, such as heart failure, cardiomyopathy, arrhythmia and the like, and have a good patent medicine prospect.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine and relates to siRNA for inhibiting PLN gene expression and its conjugate and application. Background Art

[0002] Heart failure (HF) is an increasingly serious global epidemic, with 64.3 million patients worldwide, of which 13 million will be in my country by 2023. Although the research on HF is deepening and clinical treatment methods are increasing (such as drug interventions such as angiotensin II or implantation of ventricular assist devices, etc.), the disability and mortality rates of HF patients are still high, and the long-term prognosis is poor, with a mortality rate of 50% within 5 years. Therefore, new treatments are urgently needed.

[0003] Dilated cardiomyopathy refers to the presence of left ventricular or biventricular dilation or systolic dysfunction in the absence of abnormal loading conditions or severe coronary artery disease sufficient to cause ventricular remodeling. The initial symptoms of dilated cardiomyopathy are atypical and may include decreased exercise tolerance, dyspnea, lower limb and abdominal edema, chest tightness, and palpitations. Some patients with dilated cardiomyopathy may not notice any symptoms at first. However, patients with dilated cardiomyopathy may develop heart failure and severe arrhythmias that are life-threatening. The causes of dilated cardiomyopathy are varied, including familial genetic factors.

[0004] Heart failure is characterized by decreased cardiac contractility, and a common feature is impaired calcium chelation in the sarcoplasmic reticulum (SR). The key factor is the Ca2+ chelation function that mediates SR calcium absorption. 2+ -ATPase (SERCA2a) activity is reduced, resulting in Ca 2+ Absorption is abnormal, accompanied by an increase in unphosphorylated phospholamban (PLN). PLN is the main regulator of SERCA2a activity and the only SERCA2a-related protein directly involved in the development of heart disease (including heart failure) to date. Dephosphorylated PLN is an inhibitor of SERCA, and phosphorylation of PLN can relieve this inhibition. In addition, PLN is also a key mediator of the β-adrenergic stimulation effect. Changes in its phosphorylation are related to functional changes in cardiac SR and contractility, and it is a key regulator of cardiac contractility. Therefore, PLN has become a highly potential nucleic acid drug target for the treatment of heart failure.

[0005] PLN is a reversibly phosphorylated SR protein that modifies global cardiac function. In many cases, ablation of PLN was able to successfully rescue the decreased cardiac function and Ca2+ observed in various similar animal models of heart failure. 2+Abnormal uptake. In heart failure or cardiomyopathy, elevated PLN is detrimental. Upregulation of PLN inhibits SERCA2a activity and reduces Ca 2+ In addition, the human R14del-PLN mutation causes the development of cardiomyopathy, with clinical features of arrhythmogenic cardiomyopathy and dilated cardiomyopathy, and may progress to heart failure and premature death. Similar results were found in mice. The R14del-PLN mutation caused dilated cardiomyopathy, resulting in PLN protein aggregation and cardiac dysfunction. Reducing the expression of PLN has become a strategy for treating heart failure. However, there is evidence that humans cannot completely lack PLN, otherwise it will cause severe dilated cardiomyopathy, and the safe tolerance level of PLN reduction is 50%. Therefore, it is necessary to develop more accurate, safer and more effective nucleic acid drugs with lower dosing frequency. Summary of the invention

[0006] The purpose of the present invention is to provide a siRNA molecule capable of inhibiting the expression of the PLN gene, in order to provide a new treatment for PLN-related diseases including heart failure, cardiomyopathy and arrhythmia.

[0007] In a first aspect, the present disclosure provides an siRNA for inhibiting PLN gene expression, the siRNA comprising a sense strand and an antisense strand, the sense strand comprising a nucleotide sequence I, the antisense strand comprising a nucleotide sequence II; each nucleotide in the nucleotide sequence I and the nucleotide sequence II is a modified or unmodified nucleotide; the nucleotide sequence I and the nucleotide sequence II are at least partially reverse-complemented to form a double-stranded region; the nucleotide sequence I is substantially identical to a first nucleotide sequence, the first nucleotide sequence is a nucleotide sequence of at least 19 nucleotides in length in the mRNA expressed by the PLN gene, preferably, the first nucleotide sequence is a nucleotide sequence of 19 to 25 nucleotides in length in the mRNA expressed by the PLN gene, such as 19, 20, 21, 22, 23, 24 or 25 nucleotides;

[0008] Preferably, the first nucleotide sequence is a nucleotide sequence of at least 19 nucleotides in length in the high activity interval of the mRNA expressed by the PLN gene, preferably a nucleotide sequence of 19 to 25 nucleotides, and the high activity interval is positions 292-357, 444-626 and 1127-1149 of the mRNA expressed by the PLN gene, preferably positions 292-314, 315-337, 318-340, 327-349, 335-357, 444-466, 476-498, 604-626 and 1127-1149;

[0009] Preferably, the mRNA expressed by the PLN gene is as shown in NCBI refseqID NM_002667.5; specifically, the mRNA sequence is as shown in SEQ ID NO: 1;

[0010] The high activity interval means that the siRNA and siRNA conjugate designed within this interval can effectively reduce the level of PLN mRNA. Figure 1 The results were divided into intervals according to whether the observed maximum inhibition rate of PLN mRNA by siRNA and siRNA conjugates fell within 40-60%, 60%-80%, or greater than 80%.

[0011] In some embodiments, in the above siRNA, the nucleotide sequence II is substantially reverse complementary, essentially reverse complementary, or completely reverse complementary to the first nucleotide sequence.

[0012] In some embodiments, in any of the above described siRNAs, the sense strand and antisense strand are the same or different in length, the sense strand is 16-23 nucleotides long, and the antisense strand is 19-26 nucleotides long. In some embodiments, the length ratio of the sense strand and antisense strand of the siRNA is 16 / 21, 21 / 23, or 19 / 24.

[0013] In some embodiments, in any of the above-mentioned siRNAs, the nucleotide sequence I comprises at least 15 consecutive nucleotides shown in any of the sequences in SEQ ID NO:2-100, such as at least 15, 16, 17, 18, 19, 20 or 21 nucleotides; preferably, the nucleotide sequence I is as shown in any of the sequences in SEQ ID NO:2-100.

[0014] In some embodiments, in any of the above siRNAs, the nucleotide sequence II comprises at least 15 consecutive nucleotides as shown in any of SEQ ID NOs: 101-199, such as at least 15, 16, 17, 18, 19, 20, 21, 22 or 23 nucleotides; preferably, the nucleotide sequence II is as shown in any of SEQ ID NOs: 101-199.

[0015] In some embodiments, in any of the above siRNAs, the siRNA comprises a sense strand and an antisense strand, which is:

[0016] 1) The sequence of the sense strand is shown in SEQ ID NO: 2, and the sequence of the antisense strand is shown in SEQ ID NO: 101;

[0017] 2) the sequence of the sense strand is shown in SEQ ID NO: 3, and the sequence of the antisense strand is shown in SEQ ID NO: 102;

[0018] 3) the sequence of the sense strand is shown in SEQ ID NO: 4, and the sequence of the antisense strand is shown in SEQ ID NO: 103;

[0019] 4) the sequence of the sense strand is shown in SEQ ID NO: 5, and the sequence of the antisense strand is shown in SEQ ID NO: 104;

[0020] 5) the sequence of the sense strand is shown in SEQ ID NO:6, and the sequence of the antisense strand is shown in SEQ ID NO:105;

[0021] 6) the sequence of the sense strand is shown in SEQ ID NO: 7, and the sequence of the antisense strand is shown in SEQ ID NO: 106;

[0022] 7) The sequence of the sense strand is shown in SEQ ID NO: 8, and the sequence of the antisense strand is shown in SEQ ID NO: 107;

[0023] 8) the sequence of the sense strand is shown in SEQ ID NO:9, and the sequence of the antisense strand is shown in SEQ ID NO:108;

[0024] 9) the sequence of the sense strand is shown in SEQ ID NO: 10, and the sequence of the antisense strand is shown in SEQ ID NO: 109;

[0025] 10) the sequence of the sense strand is shown in SEQ ID NO: 11, and the sequence of the antisense strand is shown in SEQ ID NO: 110;

[0026] 11) The sequence of the sense strand is shown in SEQ ID NO: 12, and the sequence of the antisense strand is shown in SEQ ID NO: 111;

[0027] 12) the sequence of the sense strand is shown in SEQ ID NO: 13, and the sequence of the antisense strand is shown in SEQ ID NO: 112;

[0028] 13) the sequence of the sense strand is shown in SEQ ID NO: 14, and the sequence of the antisense strand is shown in SEQ ID NO: 113;

[0029] 14) the sequence of the sense strand is shown in SEQ ID NO: 15, and the sequence of the antisense strand is shown in SEQ ID NO: 114;

[0030] 15) the sequence of the sense strand is shown in SEQ ID NO: 16, and the sequence of the antisense strand is shown in SEQ ID NO: 115;

[0031] 16) the sequence of the sense strand is shown in SEQ ID NO: 17, and the sequence of the antisense strand is shown in SEQ ID NO: 116;

[0032] 17) The sequence of the sense strand is shown in SEQ ID NO: 18, and the sequence of the antisense strand is shown in SEQ ID NO: 117;

[0033] 18) The sequence of the sense strand is shown in SEQ ID NO: 19, and the sequence of the antisense strand is shown in SEQ ID NO: 118;

[0034] 19) The sequence of the sense strand is shown in SEQ ID NO: 20, and the sequence of the antisense strand is shown in SEQ ID NO: 119;

[0035] 20) the sequence of the sense strand is shown in SEQ ID NO: 21, and the sequence of the antisense strand is shown in SEQ ID NO: 120;

[0036] 21) the sequence of the sense strand is shown in SEQ ID NO: 22, and the sequence of the antisense strand is shown in SEQ ID NO: 121;

[0037] 22) the sequence of the sense strand is shown in SEQ ID NO: 23, and the sequence of the antisense strand is shown in SEQ ID NO: 122;

[0038] 23) the sequence of the sense strand is shown in SEQ ID NO: 24, and the sequence of the antisense strand is shown in SEQ ID NO: 123;

[0039] 24) the sequence of the sense strand is shown in SEQ ID NO: 25, and the sequence of the antisense strand is shown in SEQ ID NO: 124;

[0040] 25) the sequence of the sense strand is shown in SEQ ID NO: 26, and the sequence of the antisense strand is shown in SEQ ID NO: 125;

[0041] 26) the sequence of the sense strand is shown in SEQ ID NO: 27, and the sequence of the antisense strand is shown in SEQ ID NO: 126;

[0042] 27) the sequence of the sense strand is shown in SEQ ID NO: 28, and the sequence of the antisense strand is shown in SEQ ID NO: 127;

[0043] 28) the sequence of the sense strand is shown in SEQ ID NO: 29, and the sequence of the antisense strand is shown in SEQ ID NO: 128;

[0044] 29) the sequence of the sense strand is shown in SEQ ID NO:30, and the sequence of the antisense strand is shown in SEQ ID NO:129;

[0045] 30) the sequence of the sense strand is shown in SEQ ID NO:31, and the sequence of the antisense strand is shown in SEQ ID NO:130;

[0046] 31) the sequence of the sense strand is shown in SEQ ID NO:32, and the sequence of the antisense strand is shown in SEQ ID NO:131;

[0047] 32) the sequence of the sense strand is shown in SEQ ID NO:33, and the sequence of the antisense strand is shown in SEQ ID NO:132;

[0048] 33) the sequence of the sense strand is shown in SEQ ID NO:34, and the sequence of the antisense strand is shown in SEQ ID NO:133;

[0049] 34) the sequence of the sense strand is shown in SEQ ID NO:35, and the sequence of the antisense strand is shown in SEQ ID NO:134;

[0050] 35) the sequence of the sense strand is shown in SEQ ID NO:36, and the sequence of the antisense strand is shown in SEQ ID NO:135;

[0051] 36) the sequence of the sense strand is shown in SEQ ID NO:37, and the sequence of the antisense strand is shown in SEQ ID NO:136;

[0052] 37) the sequence of the sense strand is shown in SEQ ID NO:38, and the sequence of the antisense strand is shown in SEQ ID NO:137;

[0053] 38) the sequence of the sense strand is shown in SEQ ID NO:39, and the sequence of the antisense strand is shown in SEQ ID NO:138;

[0054] 39) the sequence of the sense strand is shown in SEQ ID NO:40, and the sequence of the antisense strand is shown in SEQ ID NO:139;

[0055] 40) the sequence of the sense strand is shown in SEQ ID NO:41, and the sequence of the antisense strand is shown in SEQ ID NO:140;

[0056] 41) The sequence of the sense strand is shown in SEQ ID NO:42, and the sequence of the antisense strand is shown in SEQ ID NO:141;

[0057] 42) the sequence of the sense strand is shown in SEQ ID NO:43, and the sequence of the antisense strand is shown in SEQ ID NO:142;

[0058] 43) the sequence of the sense strand is shown in SEQ ID NO:44, and the sequence of the antisense strand is shown in SEQ ID NO:143;

[0059] 44) the sequence of the sense strand is shown in SEQ ID NO:45, and the sequence of the antisense strand is shown in SEQ ID NO:144;

[0060] 45) the sequence of the sense strand is shown in SEQ ID NO:46, and the sequence of the antisense strand is shown in SEQ ID NO:145;

[0061] 46) the sequence of the sense strand is shown in SEQ ID NO:47, and the sequence of the antisense strand is shown in SEQ ID NO:146;

[0062] 47) the sequence of the sense strand is shown in SEQ ID NO:48, and the sequence of the antisense strand is shown in SEQ ID NO:147;

[0063] 48) The sequence of the sense strand is shown in SEQ ID NO: 49, and the sequence of the antisense strand is shown in SEQ ID NO: 148;

[0064] 49) The sequence of the sense strand is shown in SEQ ID NO:50, and the sequence of the antisense strand is shown in SEQ ID NO:149;

[0065] 50) the sequence of the sense strand is shown in SEQ ID NO:51, and the sequence of the antisense strand is shown in SEQ ID NO:150;

[0066] 51) The sequence of the sense strand is shown in SEQ ID NO:52, and the sequence of the antisense strand is shown in SEQ ID NO:151;

[0067] 52) the sequence of the sense strand is shown in SEQ ID NO:53, and the sequence of the antisense strand is shown in SEQ ID NO:152;

[0068] 53) the sequence of the sense strand is shown in SEQ ID NO:54, and the sequence of the antisense strand is shown in SEQ ID NO:153;

[0069] 54) the sequence of the sense strand is shown in SEQ ID NO:55, and the sequence of the antisense strand is shown in SEQ ID NO:154;

[0070] 55) the sequence of the sense strand is shown in SEQ ID NO:56, and the sequence of the antisense strand is shown in SEQ ID NO:155;

[0071] 56) The sequence of the sense strand is shown in SEQ ID NO:57, and the sequence of the antisense strand is shown in SEQ ID NO:156;

[0072] 57) the sequence of the sense strand is shown in SEQ ID NO:58, and the sequence of the antisense strand is shown in SEQ ID NO:157;

[0073] 58) The sequence of the sense strand is shown in SEQ ID NO:59, and the sequence of the antisense strand is shown in SEQ ID NO:158;

[0074] 59) The sequence of the sense strand is shown in SEQ ID NO:60, and the sequence of the antisense strand is shown in SEQ ID NO:159;

[0075] 60) The sequence of the sense strand is shown in SEQ ID NO:61, and the sequence of the antisense strand is shown in SEQ ID NO:160;

[0076] 61) The sequence of the sense strand is shown in SEQ ID NO:62, and the sequence of the antisense strand is shown in SEQ ID NO:161;

[0077] 62) The sequence of the sense strand is shown in SEQ ID NO: 63, and the sequence of the antisense strand is shown in SEQ ID NO: 162;

[0078] 63) The sequence of the sense strand is shown in SEQ ID NO: 64, and the sequence of the antisense strand is shown in SEQ ID NO: 163;

[0079] 64) The sequence of the sense strand is shown in SEQ ID NO: 65, and the sequence of the antisense strand is shown in SEQ ID NO: 164;

[0080] 65) The sequence of the sense strand is shown in SEQ ID NO: 66, and the sequence of the antisense strand is shown in SEQ ID NO: 165;

[0081] 66) The sequence of the sense strand is shown in SEQ ID NO: 67, and the sequence of the antisense strand is shown in SEQ ID NO: 166;

[0082] 67) The sequence of the sense strand is shown in SEQ ID NO: 68, and the sequence of the antisense strand is shown in SEQ ID NO: 167;

[0083] 68) The sequence of the sense strand is shown in SEQ ID NO: 69, and the sequence of the antisense strand is shown in SEQ ID NO: 168;

[0084] 69) The sequence of the sense strand is shown in SEQ ID NO: 70, and the sequence of the antisense strand is shown in SEQ ID NO: 169;

[0085] 70) The sequence of the sense strand is shown in SEQ ID NO:71, and the sequence of the antisense strand is shown in SEQ ID NO:170;

[0086] 71) The sequence of the sense strand is shown in SEQ ID NO: 72, and the sequence of the antisense strand is shown in SEQ ID NO: 171;

[0087] 72) the sequence of the sense strand is shown in SEQ ID NO: 73, and the sequence of the antisense strand is shown in SEQ ID NO: 172;

[0088] 73) the sequence of the sense strand is shown in SEQ ID NO: 74, and the sequence of the antisense strand is shown in SEQ ID NO: 173;

[0089] 74) the sequence of the sense strand is shown in SEQ ID NO: 75, and the sequence of the antisense strand is shown in SEQ ID NO: 174;

[0090] 75) the sequence of the sense strand is shown in SEQ ID NO: 76, and the sequence of the antisense strand is shown in SEQ ID NO: 175;

[0091] 76) The sequence of the sense strand is shown in SEQ ID NO: 77, and the sequence of the antisense strand is shown in SEQ ID NO: 176;

[0092] 77) The sequence of the sense strand is shown in SEQ ID NO: 78, and the sequence of the antisense strand is shown in SEQ ID NO: 177;

[0093] 78) The sequence of the sense strand is shown in SEQ ID NO: 79, and the sequence of the antisense strand is shown in SEQ ID NO: 178;

[0094] 79) The sequence of the sense strand is shown in SEQ ID NO:80, and the sequence of the antisense strand is shown in SEQ ID NO:179;

[0095] 80) The sequence of the sense strand is shown in SEQ ID NO:81, and the sequence of the antisense strand is shown in SEQ ID NO:180;

[0096] 81) The sequence of the sense strand is shown in SEQ ID NO:82, and the sequence of the antisense strand is shown in SEQ ID NO:181;

[0097] 82) the sequence of the sense strand is shown in SEQ ID NO:83, and the sequence of the antisense strand is shown in SEQ ID NO:182;

[0098] 83) The sequence of the sense strand is shown in SEQ ID NO:84, and the sequence of the antisense strand is shown in SEQ ID NO:183;

[0099] 84) the sequence of the sense strand is shown in SEQ ID NO:85, and the sequence of the antisense strand is shown in SEQ ID NO:184;

[0100] 85) the sequence of the sense strand is shown in SEQ ID NO:86, and the sequence of the antisense strand is shown in SEQ ID NO:185;

[0101] 86) The sequence of the sense strand is shown in SEQ ID NO: 87, and the sequence of the antisense strand is shown in SEQ ID NO: 186;

[0102] 87) The sequence of the sense strand is shown in SEQ ID NO:88, and the sequence of the antisense strand is shown in SEQ ID NO:187;

[0103] 88) The sequence of the sense strand is shown in SEQ ID NO:89, and the sequence of the antisense strand is shown in SEQ ID NO:188;

[0104] 89) The sequence of the sense strand is shown in SEQ ID NO:90, and the sequence of the antisense strand is shown in SEQ ID NO:189;

[0105] 90) The sequence of the sense strand is shown in SEQ ID NO:91, and the sequence of the antisense strand is shown in SEQ ID NO:190;

[0106] 91) The sequence of the sense strand is shown in SEQ ID NO:92, and the sequence of the antisense strand is shown in SEQ ID NO:191;

[0107] 92) the sequence of the sense strand is shown in SEQ ID NO:93, and the sequence of the antisense strand is shown in SEQ ID NO:192;

[0108] 93) the sequence of the sense strand is shown in SEQ ID NO:94, and the sequence of the antisense strand is shown in SEQ ID NO:193;

[0109] 94) the sequence of the sense strand is shown in SEQ ID NO:95, and the sequence of the antisense strand is shown in SEQ ID NO:194;

[0110] 95) the sequence of the sense strand is shown in SEQ ID NO:96, and the sequence of the antisense strand is shown in SEQ ID NO:195;

[0111] 96) the sequence of the sense strand is shown in SEQ ID NO: 97, and the sequence of the antisense strand is shown in SEQ ID NO: 196;

[0112] 97) The sequence of the sense strand is shown in SEQ ID NO: 98, and the sequence of the antisense strand is shown in SEQ ID NO: 197;

[0113] 98) the sequence of the sense strand is shown in SEQ ID NO:99, and the sequence of the antisense strand is shown in SEQ ID NO:198; or

[0114] 99) The sequence of the sense strand is shown in SEQ ID NO: 100, and the sequence of the antisense strand is shown in SEQ ID NO: 199.

[0115] In some embodiments, in any of the above-mentioned siRNAs, each nucleotide in the nucleotide sequence I and the nucleotide sequence II is a modified nucleotide, and the modified nucleotide is a fluorinated modified nucleotide or a non-fluorinated modified nucleotide.

[0116] In some embodiments, the fluoro-modified nucleotide refers to a nucleotide in which the hydroxyl group at the 2'-position of the ribose group of the nucleotide is replaced by fluorine, and has a structure shown in the following formula (1); the non-fluoro-modified nucleotide refers to a nucleotide or nucleotide analog in which the hydroxyl group at the 2'-position of the ribose group of the nucleotide is replaced by a non-fluoro group. In some embodiments, each non-fluoro-modified nucleotide is independently selected from one of the nucleotides or nucleotide analogs in which the hydroxyl group at the 2'-position of the ribose group of the nucleotide is replaced by a non-fluoro group. These nucleotides or nucleotide analogs in which the hydroxyl group at the 2'-position of the ribose group is replaced by a non-fluoro group are well known to those skilled in the art, and these nucleotides or nucleotide analogs can be selected from one of 2'-alkoxy-modified nucleotides or nucleotide analogs, 2'-substituted alkoxy-modified nucleotides or nucleotide analogs, 2'-alkyl-modified nucleotides or nucleotide analogs, 2'-substituted alkyl-modified nucleotides or nucleotide analogs, 2'-amino-modified nucleotides or nucleotide analogs, 2'-substituted amino-modified nucleotides or nucleotide analogs, and 2'-deoxynucleotides. In some embodiments, the 2'-alkoxy modified nucleotide is a methoxy modified nucleotide (2'-OMe), as shown in formula (2). In some embodiments, the 2'-substituted alkoxy modified nucleotide, for example, can be a 2'-O-methoxyethyl modified nucleotide (2'-MOE), as shown in formula (3). In some embodiments, the 2'-amino modified nucleotide (2'-NH 2 ) is shown in formula (4). In some embodiments, the 2'-deoxynucleotide (DNA) is shown in formula (5).

[0117]

[0118] Nucleotide analogs refer to groups that can replace nucleotides in nucleic acids but have structures different from adenine ribonucleoside, guanine ribonucleotide, cytosine ribonucleotide, uracil ribonucleotide or thymine ribonucleotide. In some embodiments, nucleotide analogs can be isonucleotides, bridged nucleotides or acyclic nucleotides.

[0119] A bridging nucleotide refers to a constrained or inaccessible nucleotide. The bridging nucleotide may contain a five-membered ring, a six-membered ring, or a seven-membered ring with a fixed C3-endosugar condensed bridge structure. In some embodiments, the bridging nucleotide may be LNA, ENA, cET BNA, etc.; wherein LNA is shown in formula (6), ENA is shown in formula (7), and cET BNA is shown in formula (8).

[0120]

[0121] Acyclic nucleotides are a type of nucleotides formed by opening the sugar ring of a nucleotide. In some embodiments, the acyclic nucleotide can be an unlocked nucleic acid (UNA) or a glycerol nucleic acid (GNA), wherein UNA is shown in formula (9) and GNA is shown in formula (10).

[0122]

[0123] In the above formula (9) and formula (10), R is selected from H, OH or alkoxy (O-alkyl).

[0124] Isonucleotides refer to compounds formed by a change in the position of a base on the ribose ring of a nucleotide. In some embodiments, an isonucleotide may be a compound formed by a base moving from the 1' position to the 2' position or the 3' position of the ribose ring, as shown in formula (11) or formula (12).

[0125]

[0126] In formula (11) and formula (12), R is selected from H, OH, F or the non-fluorine group as described above.

[0127] In formula (1) to formula (12), Base represents a base.

[0128] In some embodiments, in any of the above-mentioned siRNAs, in the direction from the 5' end to the 3' end, one or more nucleotides at positions 7, 9, 10, and 11 of the nucleotide sequence I are fluorinated modified nucleotides; and, in the direction from the 5' end to the 3' end, one or more nucleotides at positions 2, 6, 8, 9, 14, and 16 of the nucleotide sequence II are fluorinated modified nucleotides.

[0129] In some embodiments, at least a portion of the phosphate groups in the phosphate-sugar backbone of at least one single strand in the sense strand and the antisense strand of the siRNA is a phosphate group with a modified group. In some embodiments, the phosphate group with a modified group is a thiophosphate group formed by replacing at least one oxygen atom in the phosphodiester bond in the phosphate group with a sulfur atom. In some embodiments, the phosphate group with a modified group is a thiophosphate group having a structure as shown in formula (13):

[0130]

[0131] In some embodiments, in any of the above siRNAs, the phosphorothioate linkage is present in at least one of the following positions: between the 1st and 2nd nucleotides of the sense strand and / or the antisense strand; between the 2nd and 3rd nucleotides of the sense strand and / or the antisense strand; between the 19th and 20th nucleotides of the antisense strand; between the 20th and 21st nucleotides of the antisense strand; between the 21st and 22nd nucleotides of the antisense strand; between the 22nd and 23rd nucleotides of the antisense strand; or any combination thereof;

[0132] In some embodiments, in any of the above siRNAs, the 5' terminal nucleotide of the antisense strand of the siRNA is a 5'-phosphate nucleotide or a 5'-phosphate analog modified nucleotide, as shown in formula (14), formula (15) and formula (16):

[0133]

[0134] In a second aspect, the present disclosure provides a siRNA conjugate, comprising any of the above-described siRNAs and a conjugated group conjugated to the siRNA. In some embodiments, the pharmaceutically acceptable conjugated group in the siRNA conjugate may be galactose or N-acetylgalactosamine, wherein the galactose or N-acetylgalactosamine molecule may be monovalent, divalent, trivalent, or tetravalent. In some embodiments, the conjugation site of the siRNA and the conjugated group may be at the 3' end or 5' end of the siRNA sense strand, or at the 3' end of the antisense strand, or in the internal sequence of the siRNA.

[0135] In some embodiments, in the above siRNA conjugate, the conjugated group is L96, and the structure is as follows:

[0136]

[0137] In some embodiments, the siRNA conjugate comprises a sense strand and an antisense strand, which is:

[0138] 1) The sequence of the sense strand is shown in SEQ ID NO: 200, and the sequence of the antisense strand is shown in SEQ ID NO: 299;

[0139] 2) the sequence of the sense strand is shown in SEQ ID NO: 201, and the sequence of the antisense strand is shown in SEQ ID NO: 300;

[0140] 3) the sequence of the sense strand is shown in SEQ ID NO: 202, and the sequence of the antisense strand is shown in SEQ ID NO: 301;

[0141] 4) the sequence of the sense strand is shown in SEQ ID NO: 203, and the sequence of the antisense strand is shown in SEQ ID NO: 302;

[0142] 5) the sequence of the sense strand is shown in SEQ ID NO: 204, and the sequence of the antisense strand is shown in SEQ ID NO: 303;

[0143] 6) the sequence of the sense strand is shown in SEQ ID NO: 205, and the sequence of the antisense strand is shown in SEQ ID NO: 304;

[0144] 7) the sequence of the sense strand is shown in SEQ ID NO: 206, and the sequence of the antisense strand is shown in SEQ ID NO: 305;

[0145] 8) the sequence of the sense strand is shown in SEQ ID NO: 207, and the sequence of the antisense strand is shown in SEQ ID NO: 306;

[0146] 9) the sequence of the sense strand is shown in SEQ ID NO: 208, and the sequence of the antisense strand is shown in SEQ ID NO: 307;

[0147] 10) the sequence of the sense strand is shown in SEQ ID NO: 209, and the sequence of the antisense strand is shown in SEQ ID NO: 308;

[0148] 11) The sequence of the sense strand is shown in SEQ ID NO: 210, and the sequence of the antisense strand is shown in SEQ ID NO: 309;

[0149] 12) the sequence of the sense strand is shown in SEQ ID NO: 211, and the sequence of the antisense strand is shown in SEQ ID NO: 310;

[0150] 13) the sequence of the sense strand is shown in SEQ ID NO: 212, and the sequence of the antisense strand is shown in SEQ ID NO: 311;

[0151] 14) the sequence of the sense strand is shown in SEQ ID NO: 213, and the sequence of the antisense strand is shown in SEQ ID NO: 312;

[0152] 15) the sequence of the sense strand is shown in SEQ ID NO: 214, and the sequence of the antisense strand is shown in SEQ ID NO: 313;

[0153] 16) the sequence of the sense strand is shown in SEQ ID NO: 215, and the sequence of the antisense strand is shown in SEQ ID NO: 314;

[0154] 17) the sequence of the sense strand is shown in SEQ ID NO: 216, and the sequence of the antisense strand is shown in SEQ ID NO: 315;

[0155] 18) the sequence of the sense strand is shown in SEQ ID NO: 217, and the sequence of the antisense strand is shown in SEQ ID NO: 316;

[0156] 19) The sequence of the sense strand is shown in SEQ ID NO: 218, and the sequence of the antisense strand is shown in SEQ ID NO: 317;

[0157] 20) the sequence of the sense strand is shown in SEQ ID NO: 219, and the sequence of the antisense strand is shown in SEQ ID NO: 318;

[0158] 21) the sequence of the sense strand is shown in SEQ ID NO: 220, and the sequence of the antisense strand is shown in SEQ ID NO: 319;

[0159] 22) the sequence of the sense strand is shown in SEQ ID NO: 221, and the sequence of the antisense strand is shown in SEQ ID NO: 320;

[0160] 23) the sequence of the sense strand is shown in SEQ ID NO: 222, and the sequence of the antisense strand is shown in SEQ ID NO: 321;

[0161] 24) the sequence of the sense strand is shown in SEQ ID NO: 223, and the sequence of the antisense strand is shown in SEQ ID NO: 322;

[0162] 25) the sequence of the sense strand is shown in SEQ ID NO: 224, and the sequence of the antisense strand is shown in SEQ ID NO: 323;

[0163] 26) the sequence of the sense strand is shown in SEQ ID NO: 225, and the sequence of the antisense strand is shown in SEQ ID NO: 324;

[0164] 27) the sequence of the sense strand is shown in SEQ ID NO: 226, and the sequence of the antisense strand is shown in SEQ ID NO: 325;

[0165] 28) the sequence of the sense strand is shown in SEQ ID NO: 227, and the sequence of the antisense strand is shown in SEQ ID NO: 326;

[0166] 29) the sequence of the sense strand is shown in SEQ ID NO: 228, and the sequence of the antisense strand is shown in SEQ ID NO: 327;

[0167] 30) the sequence of the sense strand is shown in SEQ ID NO: 229, and the sequence of the antisense strand is shown in SEQ ID NO: 328;

[0168] 31) the sequence of the sense strand is shown in SEQ ID NO: 230, and the sequence of the antisense strand is shown in SEQ ID NO: 329;

[0169] 32) the sequence of the sense strand is shown in SEQ ID NO: 231, and the sequence of the antisense strand is shown in SEQ ID NO: 330;

[0170] 33) the sequence of the sense strand is shown in SEQ ID NO: 232, and the sequence of the antisense strand is shown in SEQ ID NO: 331;

[0171] 34) the sequence of the sense strand is shown in SEQ ID NO: 233, and the sequence of the antisense strand is shown in SEQ ID NO: 332;

[0172] 35) the sequence of the sense strand is shown in SEQ ID NO: 234, and the sequence of the antisense strand is shown in SEQ ID NO: 333;

[0173] 36) the sequence of the sense strand is shown in SEQ ID NO: 235, and the sequence of the antisense strand is shown in SEQ ID NO: 334;

[0174] 37) the sequence of the sense strand is shown in SEQ ID NO: 236, and the sequence of the antisense strand is shown in SEQ ID NO: 335;

[0175] 38) the sequence of the sense strand is shown in SEQ ID NO: 237, and the sequence of the antisense strand is shown in SEQ ID NO: 336;

[0176] 39) the sequence of the sense strand is shown in SEQ ID NO: 238, and the sequence of the antisense strand is shown in SEQ ID NO: 337;

[0177] 40) the sequence of the sense strand is shown in SEQ ID NO: 239, and the sequence of the antisense strand is shown in SEQ ID NO: 338;

[0178] 41) The sequence of the sense strand is shown in SEQ ID NO: 240, and the sequence of the antisense strand is shown in SEQ ID NO: 339;

[0179] 42) the sequence of the sense strand is shown in SEQ ID NO: 241, and the sequence of the antisense strand is shown in SEQ ID NO: 340;

[0180] 43) the sequence of the sense strand is shown in SEQ ID NO: 242, and the sequence of the antisense strand is shown in SEQ ID NO: 341;

[0181] 44) the sequence of the sense strand is shown in SEQ ID NO: 243, and the sequence of the antisense strand is shown in SEQ ID NO: 342;

[0182] 45) the sequence of the sense strand is shown in SEQ ID NO: 244, and the sequence of the antisense strand is shown in SEQ ID NO: 343;

[0183] 46) the sequence of the sense strand is shown in SEQ ID NO: 245, and the sequence of the antisense strand is shown in SEQ ID NO: 344;

[0184] 47) the sequence of the sense strand is shown in SEQ ID NO: 246, and the sequence of the antisense strand is shown in SEQ ID NO: 345;

[0185] 48) The sequence of the sense strand is shown in SEQ ID NO: 247, and the sequence of the antisense strand is shown in SEQ ID NO: 346;

[0186] 49) The sequence of the sense strand is shown in SEQ ID NO: 248, and the sequence of the antisense strand is shown in SEQ ID NO: 347;

[0187] 50) the sequence of the sense strand is shown in SEQ ID NO: 249, and the sequence of the antisense strand is shown in SEQ ID NO: 348;

[0188] 51) The sequence of the sense strand is shown in SEQ ID NO: 250, and the sequence of the antisense strand is shown in SEQ ID NO: 349;

[0189] 52) the sequence of the sense strand is shown in SEQ ID NO: 251, and the sequence of the antisense strand is shown in SEQ ID NO: 350;

[0190] 53) the sequence of the sense strand is shown in SEQ ID NO: 252, and the sequence of the antisense strand is shown in SEQ ID NO: 351;

[0191] 54) the sequence of the sense strand is shown in SEQ ID NO: 253, and the sequence of the antisense strand is shown in SEQ ID NO: 352;

[0192] 55) the sequence of the sense strand is shown in SEQ ID NO: 254, and the sequence of the antisense strand is shown in SEQ ID NO: 353;

[0193] 56) The sequence of the sense strand is shown in SEQ ID NO: 255, and the sequence of the antisense strand is shown in SEQ ID NO: 354;

[0194] 57) The sequence of the sense strand is shown in SEQ ID NO: 256, and the sequence of the antisense strand is shown in SEQ ID NO: 355;

[0195] 58) The sequence of the sense strand is shown in SEQ ID NO: 257, and the sequence of the antisense strand is shown in SEQ ID NO: 356;

[0196] 59) The sequence of the sense strand is shown in SEQ ID NO: 258, and the sequence of the antisense strand is shown in SEQ ID NO: 357;

[0197] 60) The sequence of the sense strand is shown in SEQ ID NO: 259, and the sequence of the antisense strand is shown in SEQ ID NO: 358;

[0198] 61) The sequence of the sense strand is shown in SEQ ID NO: 260, and the sequence of the antisense strand is shown in SEQ ID NO: 359;

[0199] 62) The sequence of the sense strand is shown in SEQ ID NO: 261, and the sequence of the antisense strand is shown in SEQ ID NO: 360;

[0200] 63) The sequence of the sense strand is shown in SEQ ID NO: 262, and the sequence of the antisense strand is shown in SEQ ID NO: 361;

[0201] 64) The sequence of the sense strand is shown in SEQ ID NO: 263, and the sequence of the antisense strand is shown in SEQ ID NO: 362;

[0202] 65) The sequence of the sense strand is shown in SEQ ID NO: 264, and the sequence of the antisense strand is shown in SEQ ID NO: 363;

[0203] 66) The sequence of the sense strand is shown in SEQ ID NO: 265, and the sequence of the antisense strand is shown in SEQ ID NO: 364;

[0204] 67) The sequence of the sense strand is shown in SEQ ID NO: 266, and the sequence of the antisense strand is shown in SEQ ID NO: 365;

[0205] 68) The sequence of the sense strand is shown in SEQ ID NO: 267, and the sequence of the antisense strand is shown in SEQ ID NO: 366;

[0206] 69) The sequence of the sense strand is shown in SEQ ID NO: 268, and the sequence of the antisense strand is shown in SEQ ID NO: 367;

[0207] 70) The sequence of the sense strand is shown in SEQ ID NO: 269, and the sequence of the antisense strand is shown in SEQ ID NO: 368;

[0208] 71) The sequence of the sense strand is shown in SEQ ID NO: 270, and the sequence of the antisense strand is shown in SEQ ID NO: 369;

[0209] 72) The sequence of the sense strand is shown in SEQ ID NO: 271, and the sequence of the antisense strand is shown in SEQ ID NO: 370;

[0210] 73) The sequence of the sense strand is shown in SEQ ID NO: 272, and the sequence of the antisense strand is shown in SEQ ID NO: 371;

[0211] 74) the sequence of the sense strand is shown in SEQ ID NO: 273, and the sequence of the antisense strand is shown in SEQ ID NO: 372;

[0212] 75) The sequence of the sense strand is shown in SEQ ID NO: 274, and the sequence of the antisense strand is shown in SEQ ID NO: 373;

[0213] 76) The sequence of the sense strand is shown in SEQ ID NO: 275, and the sequence of the antisense strand is shown in SEQ ID NO: 374;

[0214] 77) The sequence of the sense strand is shown in SEQ ID NO: 276, and the sequence of the antisense strand is shown in SEQ ID NO: 375;

[0215] 78) The sequence of the sense strand is shown in SEQ ID NO: 277, and the sequence of the antisense strand is shown in SEQ ID NO: 376;

[0216] 79) The sequence of the sense strand is shown in SEQ ID NO: 278, and the sequence of the antisense strand is shown in SEQ ID NO: 377;

[0217] 80) The sequence of the sense strand is shown in SEQ ID NO: 279, and the sequence of the antisense strand is shown in SEQ ID NO: 378;

[0218] 81) The sequence of the sense strand is shown in SEQ ID NO: 280, and the sequence of the antisense strand is shown in SEQ ID NO: 379;

[0219] 82) the sequence of the sense strand is shown in SEQ ID NO: 281, and the sequence of the antisense strand is shown in SEQ ID NO: 380;

[0220] 83) The sequence of the sense strand is shown in SEQ ID NO: 282, and the sequence of the antisense strand is shown in SEQ ID NO: 381;

[0221] 84) the sequence of the sense strand is shown in SEQ ID NO: 283, and the sequence of the antisense strand is shown in SEQ ID NO: 382;

[0222] 85) The sequence of the sense strand is shown in SEQ ID NO: 284, and the sequence of the antisense strand is shown in SEQ ID NO: 383;

[0223] 86) The sequence of the sense strand is shown in SEQ ID NO: 285, and the sequence of the antisense strand is shown in SEQ ID NO: 384;

[0224] 87) The sequence of the sense strand is shown in SEQ ID NO: 286, and the sequence of the antisense strand is shown in SEQ ID NO: 385;

[0225] 88) The sequence of the sense strand is shown in SEQ ID NO: 287, and the sequence of the antisense strand is shown in SEQ ID NO: 386;

[0226] 89) The sequence of the sense strand is shown in SEQ ID NO: 288, and the sequence of the antisense strand is shown in SEQ ID NO: 387;

[0227] 90) The sequence of the sense strand is shown in SEQ ID NO: 289, and the sequence of the antisense strand is shown in SEQ ID NO: 388;

[0228] 91) The sequence of the sense strand is shown in SEQ ID NO: 290, and the sequence of the antisense strand is shown in SEQ ID NO: 389;

[0229] 92) the sequence of the sense strand is shown in SEQ ID NO: 291, and the sequence of the antisense strand is shown in SEQ ID NO: 390;

[0230] 93) the sequence of the sense strand is shown in SEQ ID NO: 292, and the sequence of the antisense strand is shown in SEQ ID NO: 391;

[0231] 94) the sequence of the sense strand is shown in SEQ ID NO: 293, and the sequence of the antisense strand is shown in SEQ ID NO: 392;

[0232] 95) the sequence of the sense strand is shown in SEQ ID NO: 294, and the sequence of the antisense strand is shown in SEQ ID NO: 393;

[0233] 96) The sequence of the sense strand is shown in SEQ ID NO: 295, and the sequence of the antisense strand is shown in SEQ ID NO: 394;

[0234] 97) The sequence of the sense strand is shown in SEQ ID NO: 296, and the sequence of the antisense strand is shown in SEQ ID NO: 395;

[0235] 98) the sequence of the sense strand is shown in SEQ ID NO: 297, and the sequence of the antisense strand is shown in SEQ ID NO: 396; or

[0236] 99) The sequence of the sense strand is shown in SEQ ID NO:298, and the sequence of the antisense strand is shown in SEQ ID NO:397.

[0237] In a third aspect, the present disclosure provides a composition comprising any of the above-mentioned siRNAs or siRNA conjugates.

[0238] In some embodiments, the composition is a pharmaceutical composition, and further comprises a pharmaceutically acceptable carrier or excipient. The pharmaceutically acceptable carrier or excipient involved in the present disclosure includes, but is not limited to, water for injection, sodium hydroxide, sodium dihydrogen phosphate monohydrate, sodium dihydrogen phosphate dihydrate, phosphoric acid, sodium chloride, potassium chloride, hydrochloric acid, anhydrous potassium dihydrogen phosphate, anhydrous disodium hydrogen phosphate, PEG2000, PEG6000, cholesterol, distearoylphosphatidylcholine, 1,2-dimyristyl glyceride, and dimethyl adipate.

[0239] In a fourth aspect, the present disclosure provides use of any of the above-described siRNAs, siRNA conjugates or pharmaceutical compositions in the preparation of a drug for preventing and / or treating a disease associated with PLN expression.

[0240] In a fifth aspect, the present disclosure provides a method for preventing and / or treating a disease associated with PLN expression, the method comprising administering the siRNA, siRNA conjugate or pharmaceutical composition of the present disclosure to a subject.

[0241] In some embodiments, the disease associated with PLN expression includes one or more of heart failure, cardiomyopathy, and arrhythmia;

[0242] Preferably, the heart failure includes one or more of acute heart failure, chronic heart failure, systolic heart failure, diastolic heart failure, left heart failure, right heart failure, and total heart failure;

[0243] Preferably, the cardiomyopathy is a hereditary cardiomyopathy, including one or more of cardiomyopathy caused by p.Arg14del, Arg9Cys (R9C) and / or Arg25Cys (R25C) gene mutations; More preferably, the cardiomyopathy is dilated cardiomyopathy, including one or more of dilated cardiomyopathy caused by TTN, LMNA, RBM20, SCN5A, MYH7, TNNT2 and / or TPM1 mutations;

[0244] Preferably, the arrhythmia comprises ventricular tachycardia and / or ventricular fibrillation.

[0245] The pharmaceutical composition disclosed herein can be used alone to treat diseases associated with PLN expression, or can be used in combination with standard oral drugs, providing experimental support for the implementation of diversified treatment options for clinical patients with the above diseases.

[0246] A typical suitable dosage range of the siRNA, siRNA conjugate or pharmaceutical composition for inhibiting PLN gene expression involved in the present disclosure will be from about 0.1 mg / kg to about 10.0 mg / kg, preferably from about 0.3 mg / kg to about 3.0 mg / kg, based on the amount of siRNA contained therein.

[0247] The administration routes involved in the present disclosure include intravenous administration, subcutaneous administration, intrathecal injection, intramuscular administration, transdermal administration, airway administration (aerosol), ocular administration, nasal administration, rectal administration, pulmonary administration and topical administration (including buccal administration and sublingual administration).

[0248] The siRNA, siRNA conjugate and pharmaceutical composition disclosed in the present invention can specifically target the heart, complementarily pair with the cardiac PLN mRNA sequence, induce PLN mRNA degradation, thereby inhibiting the protein synthesis and aggregation of PLN in the heart, and improving the sarcoplasmic reticulum's ability to respond to Ca 2+ normal uptake and cardiac contractility while avoiding off-target effects.

[0249] In addition, compared with traditional small molecule drugs and antibody drugs, small nucleic acid drugs can directly regulate upstream gene expression and are relatively less likely to develop drug resistance. Moreover, small nucleic acid drugs have a long half-life in the body, so the frequency of administration is low (the drug can be given once every six months), and patient compliance is good.

[0250] In summary, the siRNA, siRNA conjugates and pharmaceutical compositions provided by the present disclosure have strong inhibitory ability on the PLN gene, can significantly reduce the expression of PLN mRNA, and have low drug toxicity. Therefore, the siRNA, siRNA conjugates and pharmaceutical compositions disclosed in the present disclosure can effectively prevent and / or treat diseases associated with PLN expression, such as heart failure, cardiomyopathy, arrhythmia, etc., provide patients with more effective, safe and convenient therapeutic drugs, and have good drug development prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0251] Figure 1 Schematic diagram of the high-efficiency activity range of PLN mRNA.

[0252] Figure 2 The results of siRNA conjugate cytotoxicity experiments.

[0253] Figure 3 The results of in vivo study on the inhibition of PLN expression by siRNA conjugates. DETAILED DESCRIPTION

[0254] definition

[0255] In the above and below, unless otherwise specified, capital letters C, G, U, A represent cytosine, guanine, uracil, and adenine nucleotides; lowercase letter m represents that the nucleotide adjacent to the left of the letter m is a methoxy-modified nucleotide; lowercase letter f represents that the nucleotide adjacent to the left of the letter f is a fluorinated-modified nucleotide; lowercase letter s represents that the two nucleotides adjacent to the left and right of the letter s are connected by thiophosphate subunits; the letter combination VP represents that the nucleotide adjacent to the right of the letter combination VP is a vinyl phosphate (5'-(E)-vinylphosphonate, E-VP)-modified nucleotide; L96 has the structure of formula (I) and is connected to the 3' end of the sense chain via a phosphate bond.

[0256] In the above and below, the "fluorinated modified nucleotide" refers to a nucleotide in which the hydroxyl group at the 2' position of the ribose group of the nucleotide is replaced by fluorine, and the "non-fluorinated modified nucleotide" refers to a nucleotide or nucleotide analog in which the hydroxyl group at the 2' position of the ribose group of the nucleotide is replaced by a non-fluorinated group. "Nucleotide analog" refers to a group that can replace nucleotides in nucleic acids but has a structure different from adenine ribonucleotides, guanine ribonucleotides, cytosine ribonucleotides, uracil ribonucleotides or thymine deoxyribonucleotides. Such as isonucleotides, bridged nucleic acids (BNA for short) or acyclic nucleotides. The "methoxy-modified nucleotide" refers to a nucleotide in which the hydroxyl group at the 2' position of the ribose group is replaced by a methoxy group. In the context of this article, the expressions "complementary" or "reverse complementary" can be used interchangeably and have the meanings well known to those skilled in the art, that is, in a double-stranded nucleic acid molecule, the bases of one chain are each paired with the bases on the other chain in a complementary manner. In DNA, the purine base adenine (A) always pairs with the pyrimidine base thymine (T) (or uracil (U) in RNA); the purine base guanine (C) always pairs with the pyrimidine base cytosine (G). Each base pair includes a purine and a pyrimidine. When adenine on one chain always pairs with thymine (or uracil) on the other chain, and guanine always pairs with cytosine, the two chains are considered to be complementary to each other, and the sequence of the chain can be inferred from the sequence of its complementary chain. Accordingly, "mismatch" in the art means that in a double-stranded nucleic acid, the bases at corresponding positions are not paired in a complementary form. In the above and below, unless otherwise specified, "substantially reverse complementary" means that there are no more than 3 base mismatches between the two nucleotide sequences involved; "substantially reverse complementary" means that there are no more than 1 base mismatch between the two nucleotide sequences; "completely reverse complementary" means that there is no base mismatch between the two nucleotide sequences. In the above and below, especially when describing the preparation method of siRNA, pharmaceutical composition or siRNA conjugate of the present disclosure, unless otherwise specified, the nucleoside monomer (nucleoside monomer) refers to the modified or unmodified nucleoside phosphoramidite monomer (unmodified or modified RNA phosphoramidites, sometimes RNA phosphoramidites are also called Nucleoside phosphora-midites) used in phosphoramidite solid phase synthesis according to the type and order of nucleotides in the siRNA or siRNA conjugate to be prepared. Phosphoramidite solid phase synthesis is a method used in siRNA synthesis known to those skilled in the art. The nucleoside monomers used in the present disclosure are all commercially available.

[0257] In Table 1 and Table 2, if there is no VP on the left side of the 5' terminal nucleotide of the sense strand and the modified sense strand connected to the conjugated group, it means that the 5' terminal nucleotide is not connected to a 5' phosphate group or a 5' phosphate derivative group, and its structure is shown in formula (II):

[0258]

[0259] Wherein, Base represents a base, such as A, U, G, C or T; R is a hydroxyl group or is substituted by various groups known to those skilled in the art, for example, R can be 2'-fluoro (2'-F), 2'-alkoxy, 2'-substituted alkoxy, 2'-alkyl, 2'-substituted alkyl, 2'-amino, 2'-substituted amino, or 2'-deoxynucleotide.

[0260] In Table 1 and Table 2, if there is no VP on the left side of the 5' terminal nucleotide of the antisense strand and the modified antisense strand, it means that the 5' terminal nucleotide is not connected to a 5' phosphate group or a 5' phosphate derivative group, and its structure is also shown in formula (II).

[0261] In Table 1 and Table 2, the 3' position of the 3' terminal nucleotide of the sense strand, the 3' terminal nucleotide of the antisense strand and the modified antisense strand is a hydroxyl group.

[0262] Example

[0263] Other objects, features and advantages of the present disclosure will become apparent from the following detailed description. However, it should be understood that the detailed description and specific examples (although representing specific embodiments of the present disclosure) are given for illustrative purposes only, because after reading the detailed description, various changes and modifications made within the spirit and scope of the present disclosure will become apparent to those skilled in the art.

[0264] The experimental techniques and experimental methods used in this example are all conventional technical methods unless otherwise specified. For example, the experimental methods in the following examples that do not specify specific conditions are usually carried out under conventional conditions such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or under conditions recommended by the manufacturer. The materials, reagents, etc. used in the examples can be obtained through regular commercial channels unless otherwise specified.

[0265] Example 1. siRNA design and synthesis

[0266] 1.1 siRNA design

[0267] OligoWalk was used to design a set of siRNA targeting human PLN gene (human PLN gene: NCBIrefseqID NM_002667.5; NCBI Gene ID: 5350). Human NM_002667.5REFSEQ mRNA has a length of 2989 bases. At the same time, in order to avoid any sequence toxicity, sequences similar to human genes should be excluded.

[0268]

[0269] 1.2 siRNA sequence synthesis

[0270] siRNAs, including negative control siRNA (siCtrl), were synthesized according to standard oligonucleotide solid phase synthesis protocols.

[0271] Oligonucleotide solid phase synthesis scheme: RNA was synthesized at a scale of 500 nmol using commercially available 5'-DMT-2'-TBDMS-rU phosphoramidite monomers, 5'-DMT-2'-TBDMS-rA (Bz) phosphoramidite monomers, 5'-DMT-2'-TBDMS-rC (Ac) phosphoramidite monomers, and 5'-DMT-2'-TBDMS-rG (iBu) phosphoramidite monomers. The phosphoramidite solution was prepared at a concentration of 50 mM, and 0.3 M benzylthiotetrazolyl (BTT) acetonitrile solution was used as an activator. During the synthesis, 0.1 M oxidizing agent (pyridine: THF: water = 20: 78: 2) was used to convert trivalent phosphorus into a pentavalent phosphorus-stabilized phosphate backbone. After the synthesis was completed, the sequence was aminolyzed from the solid phase support and precipitated. The 2'O-tert-butyldimethylsilyl protecting group of 2' was removed with triethylamine trihydrofluoric acid.

[0272] For the RNA sequence that has been synthesized, use ammonia water: methylamine = 1:1 aminolysis solution for 40 minutes at 55°C. After the aminolysis is completed, remove the solid phase carrier and take the supernatant to dryness. Add triethylamine trihydrofluoric acid: triethylamine: NMP = 6:4:3 protective group removal agent, react at 60°C for 2 hours, add n-butanol at a ratio of 1:5, stand at -20°C for 30 minutes, and centrifuge to obtain the precipitate. Add RNase-free water to dissolve, and purify by reverse phase chromatography (0.1M triethylamine acetic acid (TEAA) and acetonitrile). The purified sample is ultrafiltered and desalted with PBS. Annealing is performed to obtain siRNA, and the obtained siRNA is verified. The results show that the target siRNA is successfully prepared.

[0273] 1.3 siRNA sequence modification and conjugate synthesis

[0274] The modified siRNA is synthesized according to the oligonucleotide solid phase synthesis scheme. The modified nucleotide group can be introduced into the siRNA of the present disclosure by using a nucleoside monomer with a corresponding modification. The method for preparing a nucleoside monomer with a corresponding modification is also well known to those skilled in the art. The siRNA conjugate is synthesized by conjugating L96 to siRNA with reference to the synthesis method disclosed in WO2014025805A1 or WO2017015109A1.

[0275] The structure of the conjugated group L96 is shown below:

[0276]

[0277] Annealing of oligoribonucleotides to produce siRNA conjugates: The RNA oligomers to be annealed are washed with sterile RNaseFree HCl. 2 O (without RNA hydrolase) was prepared into a 200 μM solution. The annealing reaction system was set up as follows: a total volume of 100 μL of the above solution (duplex concentration of 10 nmol) was placed in a 95°C water bath for 10 minutes (≥100 nmol required high temperature for 20 minutes) → quickly placed in a 60°C water bath to cool naturally → the annealed solution was stored at 4°C. The complementary chains were mixed by combining equimolar RNA solutions. After identification, the siRNA conjugate was constructed correctly.

[0278] The siRNA and siRNA conjugate solutions were prepared into dry powder for later use.

[0279] The sequences of the synthesized siRNAs are shown in Table 1 below, and the schematic diagram of the high-efficiency activity range of PLN mRNA is shown in Figure 1 shown.

[0280] Table 1. List of siRNA sequences targeting PLN

[0281]

[0282]

[0283]

[0284]

[0285] Wherein, A, U, G, C represent adenine, uracil, guanine, and cytosine nucleotides. The sequences of the synthesized siRNA conjugates are shown in Table 2 below:

[0286] Table 2: siRNA conjugates targeting PLN

[0287]

[0288]

[0289]

[0290]

[0291]

[0292]

[0293] Among them, the lowercase letter m indicates that the nucleotide adjacent to the left of the letter m is a methoxy-modified nucleotide; the lowercase letter f indicates that the nucleotide adjacent to the left of the letter f is a fluorinated-modified nucleotide; the lowercase letter s indicates that the two nucleotides adjacent to the left and right of the letter s are connected by thiophosphate subunits; VP indicates that the nucleotide adjacent to the right of the letter combination VP is a vinyl phosphate (5'-(E)-vinylphosphonate, E-VP) modified nucleotide; L96 represents the L96 conjugated group connected to siRNA.

[0294] Example 2. In vitro activity screening of siRNA 293T cell line

[0295] 2.1 Experimental procedures

[0296] 2.1.1 Cell culture

[0297] 293T cells (China Center for Type Culture Collection (CCTCC), catalog number: CVCL_0063) were cultured at 37°C and 5% CO 2 The cells were cultured in a complete DMEM medium (Eallbio, supplemented with 10% FBS) and when the confluence rate reached 80%-90%, the cells were trypsinized, counted and transfected.

[0298] 2.1.2 Preparation of siRNA dilution

[0299] (1) The dry powder of the siRNA to be tested is centrifuged at low temperature and high speed, and then dissolved in ultrapure distilled water to prepare a 100 μM siRNA stock solution.

[0300] (2) Prepare 200 nM siRNA dilution Y

[0301] a) taking 50 μl of the 100 μM siRNA stock solution prepared in step (1) above, adding 50 μl of ultrapure distilled water to obtain a siRNA dilution solution with a final concentration of 50 μM;

[0302] b) taking 2 μl of the 50 μM siRNA dilution prepared in step a), adding 18 μl of ultrapure distilled water to obtain a siRNA stock solution X with a final concentration of 5 μM;

[0303] c) Take 2 μl of the prepared siRNA stock solution X, add 48 μl Opti-medium (Opti-MEM I Medium, Gibco, catalog number: 31985070) to obtain 200 nM siRNA dilution solution Y.

[0304] 2.1.3 293T cell transfection

[0305] Pick RNAiMAX transfection reagent (Thermo Fisher, catalog number: 13778150) 0.6 μl, add 10 μl Opti-medium, and obtain RNAiMAX transfection reagent diluent; RNAiMAX transfection reagent diluent and 200nM siRNA diluent Y prepared in 2.1.2 were mixed in a volume ratio of 1:1 to prepare a transfection mixture, which was allowed to stand for 5 minutes. 10μl of the transfection mixture was added to a 96-well plate, and 90μl of 293T cells cultured in 2.1.1 were added (final volume 100μl / well, the number of cells was 20,000 / well, and the concentration of siRNA in this system was 10nM); the above transfection was followed by culture for 24 hours.

[0306] 2.1.4 RNA extraction

[0307] According to the FlysisAmp Cells-to-CT 1-Step SYBR Green Kit (Nanjing Novozymes Biotech Co., Ltd., Catalog No.: CL132-01) product manual, the total RNA of the 293T cells obtained in 2.1.3 was extracted.

[0308] 2.1.5 Fluorescence quantitative PCR

[0309] The extracted total RNA was reverse transcribed and analyzed by real-time PCR using the FlysisAmp Cells-to-CT 1-Step SYBR Green Kit.

[0310] 2.1.6 Result Analysis

[0311] (1) The Ct value was automatically calculated using the software of the 7500 real-time fluorescence quantitative PCR instrument (Thermo Fisher);

[0312] (2) The relative expression of genes was calculated using the following formula:

[0313] ΔCt1=Ct(PLN group)–Ct(ACTIN of PLN group)

[0314] ΔCt2=Ct(siCtrl group)–Ct(ACTIN of siCtrl group)

[0315] ΔΔCt = ΔCt1-ΔCt2, where siCtrl group represents nonspecific siRNA sequence and is the negative control group; mRNA expression relative to siCtrl group = 2 -ΔΔCt

[0316] Inhibition rate (%) = (1-mRNA expression relative to the siCtrl group) × 100%.

[0317] 2.2 Experimental Results

[0318] The inhibitory effects of the siRNA disclosed in the present invention are shown in Table 3 below:

[0319] Table 3. Results of in vitro screening of siRNA 293T cell lines

[0320]

[0321]

[0322]

[0323] It can be seen from Table 3 that some siRNAs disclosed in the present invention can significantly inhibit the expression of PLN gene at 10 nM.

[0324] Example 3. Cytotoxicity Experiment

[0325] 3.1 Experimental procedures

[0326] 3.1.1 Cell culture

[0327] Incubate 293T cells at 37°C with 5% CO 2 The cells were cultured in a complete DMEM medium (Eallbio, catalog number: 03.1002C; supplemented with 10% FBS) in an environment. When the confluence rate reached 80%-90%, the cells were trypsinized, counted and transfected.

[0328] 3.1.2 293T cell transfection

[0329] Based on the results of Example 2 above, some siRNAs with good in vitro activity were screened for toxicity determination. 293T cells were transfected in a manner similar to Example 2, and the cytotoxicity of each siRNA conjugate was measured by determining the ratio of cell viability / toxicity in each sample after 72 hours of culture, wherein the concentrations of the transfected siRNA conjugates were 50 nM and 5 nM, respectively. Cell viability was measured by determining the intracellular ATP content using the CellTiter-Glo assay according to the manufacturer's protocol. Cytotoxicity in the supernatant was measured using ToxiLightTM according to the manufacturer's protocol. Cytotoxicity results are shown in Figure 2 .

[0330] 3.2 Experimental Results

[0331] The results show that some of the siRNA conjugates disclosed in the present invention have low cytotoxicity and good cell compatibility.

[0332] Based on the in vitro screening results of Example 2 and Example 3, the highly effective activity intervals for inhibiting PLN mRNA were determined to be positions 292-357, 444-626, and 1127-1149 calculated according to NCBI refseqID NM_002667.5.

[0333] Example 4. In vivo activity screening of siRNA conjugates

[0334] According to the in vitro screening results of Examples 2 and 3, some siRNA sequences with good in vitro activity were selected to verify the in vivo activity of the siRNA conjugates.

[0335] 4.1 Experimental procedures

[0336] 6-8 week old male mice (C57BL / 6) were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd. Each mouse was intravenously injected with 1×10 11 A recombinant adeno-associated virus 8 (AAV8) vector containing 100 genome copies. The recombinant AAV8 vector was packaged by an AAV8 capsid protein expression plasmid and a transfer plasmid carrying AAV8-hPLN, wherein the transfer plasmid carried the 354-2989 position of the human PLN sequence (NM_002667.5), which was controlled by the TBG promoter (AAV8-TBG-hPLN), and was purchased from Yunzhou Biotechnology (Guangzhou) Co., Ltd. The AAV8-hPLN transgenic mouse model was constructed 14 days after injection. Then, each mouse was given subcutaneous administration at a dose of 3 mg / kg of the conjugate. Mice were killed on the 7th day (D7), the 14th day (D14), and the 19th day (D19) after administration, with 6 mice in each group, and liver tissues were taken for PLN mRNA expression detection. Total RNA was extracted by Trizol method, mRNA was reverse transcribed using HiScript III RT SuperMix for qPCR (+gDNA wiper) kit (Vazyme, catalog number: R323-01), and real-time fluorescence quantitative PCR was performed using ChamQ Universal SYBR qPCR Master Mix kit (Vazyme, catalog number: Q711-03). The results of in vivo activity screening of siRNA conjugates are shown in Tables 4 and 5.

[0337] Table 4. In vivo screening results of siRNA conjugates

[0338]

[0339]

[0340] Table 5. In vivo screening results of siRNA conjugates

[0341]

[0342] 4.2 Experimental Results

[0343] Some chemically modified siRNA sequences have good activity in mice. YGND11-13M, YGND11-16M, YGND11-17M, YGND11-18M, YGND11-21M, YGND11-26M, YGND11-31M, YGND11-41M and YGND11-73M can reduce the expression level of PLN in mice, among which YGND11-13M, YGND11-21M, YGND11-31M, YGND11-41M and YGND11-73M have high efficiency in knocking down PLN gene expression.

[0344] Example 5. Screening of multiple doses of siRNA conjugates for in vivo activity

[0345] Based on the in vivo experimental screening results of Example 4 above, some siRNA conjugates were selected for in vivo activity verification.

[0346] 5.1 Experimental procedures

[0347] The AAV8-hPLN transgenic mouse model was constructed using the method described in Example 4. The conjugate was administered subcutaneously at a dose of 0.3 mg / kg, 1 mg / kg, and 3 mg / kg per mouse. The mice were killed on the 19th day after administration, and liver tissue was taken for PLN mRNA expression detection. The detection method and reagents were the same as in Example 4. The results of the in vivo activity screening of the siRNA conjugates at multiple doses are shown in Tables 6 and Figure 3 shown.

[0348] Table 6. Results of multiple dose in vivo screening of siRNA conjugates

[0349]

[0350]

[0351] Note: N / A means data not measured

[0352] 5.2 Experimental Results

[0353] The results showed that the siRNA conjugates disclosed in the present invention all showed good dose-dependent effects, among which YGND11-21M had the highest inhibition efficiency on PLN gene expression, with an inhibition efficiency of up to 71.57% at a dosage of 3 mg / kg. In addition, YGND11-26M had an inhibition efficiency of up to 65.27% at a dosage of 3 mg / kg.

Claims

1. An siRNA for inhibiting PLN gene expression, the siRNA comprising a sense strand and an antisense strand, the sense strand comprising a nucleotide sequence I, the antisense strand comprising a nucleotide sequence II; each nucleotide in the nucleotide sequence I and the nucleotide sequence II is a modified or unmodified nucleotide; the nucleotide sequence I and the nucleotide sequence II are at least partially reverse-complemented to form a double-stranded region; the nucleotide sequence I is substantially identical to a first nucleotide sequence, the first nucleotide sequence is a nucleotide sequence of at least 19 nucleotides in length in the mRNA expressed by the PLN gene, preferably, the first nucleotide sequence is a nucleotide sequence of 19 to 25 nucleotides in length in the mRNA expressed by the PLN gene, such as 19, 20, 21, 22, 23, 24 or 25 nucleotides; Preferably, the first nucleotide sequence is a nucleotide sequence of at least 19 nucleotides in length in the high activity interval of the mRNA expressed by the PLN gene, preferably a nucleotide sequence of 19 to 25 nucleotides, and the high activity interval is positions 292-357, 444-626 and 1127-1149 of the mRNA expressed by the PLN gene, preferably positions 292-314, 315-337, 318-340, 327-349, 335-357, 444-466, 476-498, 604-626 and 1127-1149; Preferably, the mRNA expressed by the PLN gene is as shown in NCBI refseqID NM_002667.5; specifically, the mRNA sequence is as shown in SEQ ID NO:

1.

2. The siRNA according to claim 1, wherein The nucleotide sequence I has at least 70%, at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the first nucleotide sequence.

3. The siRNA according to claim 1 or 2, wherein The nucleotide sequence I comprises at least 15 consecutive nucleotides as shown in any one of SEQ ID NOs: 2-100, such as at least 15, 16, 17, 18, 19, 20 or 21 nucleotides; preferably, the nucleotide sequence I is as shown in any one of SEQ ID NOs: 2-100; The nucleotide sequence II comprises at least 15 consecutive nucleotides as shown in any one of SEQ ID NOs: 101-199, such as at least 15, 16, 17, 18, 19, 20, 21, 22 or 23 nucleotides; preferably, the nucleotide sequence II is as shown in any one of SEQ ID NOs: 101-199.

4. The siRNA according to any one of claims 1 to 3, wherein the siRNA comprises a sense strand and an antisense strand, which is: 1) The sequence of the sense strand is shown in SEQ ID NO: 2, and the sequence of the antisense strand is shown in SEQ ID NO: 101; 2) the sequence of the sense strand is shown in SEQ ID NO: 3, and the sequence of the antisense strand is shown in SEQ ID NO: 102; 3) the sequence of the sense strand is shown in SEQ ID NO: 4, and the sequence of the antisense strand is shown in SEQ ID NO: 103; 4) the sequence of the sense strand is shown in SEQ ID NO: 5, and the sequence of the antisense strand is shown in SEQ ID NO: 104; 5) the sequence of the sense strand is shown in SEQ ID NO:6, and the sequence of the antisense strand is shown in SEQ ID NO:105; 6) the sequence of the sense strand is shown in SEQ ID NO: 7, and the sequence of the antisense strand is shown in SEQ ID NO: 106; 7) The sequence of the sense strand is shown in SEQ ID NO: 8, and the sequence of the antisense strand is shown in SEQ ID NO: 107; 8) the sequence of the sense strand is shown in SEQ ID NO:9, and the sequence of the antisense strand is shown in SEQ ID NO:108; 9) the sequence of the sense strand is shown in SEQ ID NO: 10, and the sequence of the antisense strand is shown in SEQ ID NO: 109; 10) the sequence of the sense strand is shown in SEQ ID NO: 11, and the sequence of the antisense strand is shown in SEQ ID NO: 110; 11) The sequence of the sense strand is shown in SEQ ID NO: 12, and the sequence of the antisense strand is shown in SEQ ID NO: 111; 12) the sequence of the sense strand is shown in SEQ ID NO: 13, and the sequence of the antisense strand is shown in SEQ ID NO: 112; 13) the sequence of the sense strand is shown in SEQ ID NO: 14, and the sequence of the antisense strand is shown in SEQ ID NO: 113; 14) the sequence of the sense strand is shown in SEQ ID NO: 15, and the sequence of the antisense strand is shown in SEQ ID NO: 114; 15) the sequence of the sense strand is shown in SEQ ID NO: 16, and the sequence of the antisense strand is shown in SEQ ID NO: 115; 16) the sequence of the sense strand is shown in SEQ ID NO: 17, and the sequence of the antisense strand is shown in SEQ ID NO: 116; 17) The sequence of the sense strand is shown in SEQ ID NO: 18, and the sequence of the antisense strand is shown in SEQ ID NO: 117; 18) The sequence of the sense strand is shown in SEQ ID NO: 19, and the sequence of the antisense strand is shown in SEQ ID NO: 118; 19) The sequence of the sense strand is shown in SEQ ID NO: 20, and the sequence of the antisense strand is shown in SEQ ID NO: 119; 20) the sequence of the sense strand is shown in SEQ ID NO: 21, and the sequence of the antisense strand is shown in SEQ ID NO: 120; 21) the sequence of the sense strand is shown in SEQ ID NO: 22, and the sequence of the antisense strand is shown in SEQ ID NO: 121; 22) the sequence of the sense strand is shown in SEQ ID NO: 23, and the sequence of the antisense strand is shown in SEQ ID NO: 122; 23) the sequence of the sense strand is shown in SEQ ID NO: 24, and the sequence of the antisense strand is shown in SEQ ID NO: 123; 24) the sequence of the sense strand is shown in SEQ ID NO: 25, and the sequence of the antisense strand is shown in SEQ ID NO: 124; 25) the sequence of the sense strand is shown in SEQ ID NO: 26, and the sequence of the antisense strand is shown in SEQ ID NO: 125; 26) the sequence of the sense strand is shown in SEQ ID NO: 27, and the sequence of the antisense strand is shown in SEQ ID NO: 126; 27) the sequence of the sense strand is shown in SEQ ID NO: 28, and the sequence of the antisense strand is shown in SEQ ID NO: 127; 28) the sequence of the sense strand is shown in SEQ ID NO: 29, and the sequence of the antisense strand is shown in SEQ ID NO: 128; 29) the sequence of the sense strand is shown in SEQ ID NO:30, and the sequence of the antisense strand is shown in SEQ ID NO:129; 30) the sequence of the sense strand is shown in SEQ ID NO:31, and the sequence of the antisense strand is shown in SEQ ID NO:130; 31) the sequence of the sense strand is shown in SEQ ID NO:32, and the sequence of the antisense strand is shown in SEQ ID NO:131; 32) the sequence of the sense strand is shown in SEQ ID NO:33, and the sequence of the antisense strand is shown in SEQ ID NO:132; 33) the sequence of the sense strand is shown in SEQ ID NO:34, and the sequence of the antisense strand is shown in SEQ ID NO:133; 34) the sequence of the sense strand is shown in SEQ ID NO:35, and the sequence of the antisense strand is shown in SEQ ID NO:134; 35) the sequence of the sense strand is shown in SEQ ID NO:36, and the sequence of the antisense strand is shown in SEQ ID NO:135; 36) the sequence of the sense strand is shown in SEQ ID NO:37, and the sequence of the antisense strand is shown in SEQ ID NO:136; 37) the sequence of the sense strand is shown in SEQ ID NO:38, and the sequence of the antisense strand is shown in SEQ ID NO:137; 38) the sequence of the sense strand is shown in SEQ ID NO:39, and the sequence of the antisense strand is shown in SEQ ID NO:138; 39) the sequence of the sense strand is shown in SEQ ID NO:40, and the sequence of the antisense strand is shown in SEQ ID NO:139; 40) the sequence of the sense strand is shown in SEQ ID NO:41, and the sequence of the antisense strand is shown in SEQ ID NO:140; 41) The sequence of the sense strand is shown in SEQ ID NO:42, and the sequence of the antisense strand is shown in SEQ ID NO:141; 42) the sequence of the sense strand is shown in SEQ ID NO:43, and the sequence of the antisense strand is shown in SEQ ID NO:142; 43) the sequence of the sense strand is shown in SEQ ID NO:44, and the sequence of the antisense strand is shown in SEQ ID NO:143; 44) the sequence of the sense strand is shown in SEQ ID NO:45, and the sequence of the antisense strand is shown in SEQ ID NO:144; 45) the sequence of the sense strand is shown in SEQ ID NO:46, and the sequence of the antisense strand is shown in SEQ ID NO:145; 46) the sequence of the sense strand is shown in SEQ ID NO:47, and the sequence of the antisense strand is shown in SEQ ID NO:146; 47) the sequence of the sense strand is shown in SEQ ID NO:48, and the sequence of the antisense strand is shown in SEQ ID NO:147; 48) The sequence of the sense strand is shown in SEQ ID NO: 49, and the sequence of the antisense strand is shown in SEQ ID NO: 148; 49) The sequence of the sense strand is shown in SEQ ID NO:50, and the sequence of the antisense strand is shown in SEQ ID NO:149; 50) the sequence of the sense strand is shown in SEQ ID NO:51, and the sequence of the antisense strand is shown in SEQ ID NO:150; 51) The sequence of the sense strand is shown in SEQ ID NO:52, and the sequence of the antisense strand is shown in SEQ ID NO:151; 52) the sequence of the sense strand is shown in SEQ ID NO:53, and the sequence of the antisense strand is shown in SEQ ID NO:152; 53) the sequence of the sense strand is shown in SEQ ID NO:54, and the sequence of the antisense strand is shown in SEQ ID NO:153; 54) the sequence of the sense strand is shown in SEQ ID NO:55, and the sequence of the antisense strand is shown in SEQ ID NO:154; 55) the sequence of the sense strand is shown in SEQ ID NO:56, and the sequence of the antisense strand is shown in SEQ ID NO:155; 56) The sequence of the sense strand is shown in SEQ ID NO:57, and the sequence of the antisense strand is shown in SEQ ID NO:156; 57) the sequence of the sense strand is shown in SEQ ID NO:58, and the sequence of the antisense strand is shown in SEQ ID NO:157; 58) The sequence of the sense strand is shown in SEQ ID NO:59, and the sequence of the antisense strand is shown in SEQ ID NO:158; 59) The sequence of the sense strand is shown in SEQ ID NO:60, and the sequence of the antisense strand is shown in SEQ ID NO:159; 60) The sequence of the sense strand is shown in SEQ ID NO:61, and the sequence of the antisense strand is shown in SEQ ID NO:160; 61) The sequence of the sense strand is shown in SEQ ID NO:62, and the sequence of the antisense strand is shown in SEQ ID NO:161; 62) The sequence of the sense strand is shown in SEQ ID NO: 63, and the sequence of the antisense strand is shown in SEQ ID NO: 162; 63) The sequence of the sense strand is shown in SEQ ID NO: 64, and the sequence of the antisense strand is shown in SEQ ID NO: 163; 64) The sequence of the sense strand is shown in SEQ ID NO: 65, and the sequence of the antisense strand is shown in SEQ ID NO: 164; 65) The sequence of the sense strand is shown in SEQ ID NO: 66, and the sequence of the antisense strand is shown in SEQ ID NO: 165; 66) The sequence of the sense strand is shown in SEQ ID NO: 67, and the sequence of the antisense strand is shown in SEQ ID NO: 166; 67) The sequence of the sense strand is shown in SEQ ID NO: 68, and the sequence of the antisense strand is shown in SEQ ID NO: 167; 68) The sequence of the sense strand is shown in SEQ ID NO: 69, and the sequence of the antisense strand is shown in SEQ ID NO: 168; 69) The sequence of the sense strand is shown in SEQ ID NO: 70, and the sequence of the antisense strand is shown in SEQ ID NO: 169; 70) The sequence of the sense strand is shown in SEQ ID NO:71, and the sequence of the antisense strand is shown in SEQ ID NO:170; 71) The sequence of the sense strand is shown in SEQ ID NO: 72, and the sequence of the antisense strand is shown in SEQ ID NO: 171; 72) the sequence of the sense strand is shown in SEQ ID NO: 73, and the sequence of the antisense strand is shown in SEQ ID NO: 172; 73) the sequence of the sense strand is shown in SEQ ID NO: 74, and the sequence of the antisense strand is shown in SEQ ID NO: 173; 74) the sequence of the sense strand is shown in SEQ ID NO: 75, and the sequence of the antisense strand is shown in SEQ ID NO: 174; 75) the sequence of the sense strand is shown in SEQ ID NO: 76, and the sequence of the antisense strand is shown in SEQ ID NO: 175; 76) The sequence of the sense strand is shown in SEQ ID NO: 77, and the sequence of the antisense strand is shown in SEQ ID NO: 176; 77) The sequence of the sense strand is shown in SEQ ID NO: 78, and the sequence of the antisense strand is shown in SEQ ID NO: 177; 78) The sequence of the sense strand is shown in SEQ ID NO: 79, and the sequence of the antisense strand is shown in SEQ ID NO: 178; 79) The sequence of the sense strand is shown in SEQ ID NO:80, and the sequence of the antisense strand is shown in SEQ ID NO:179; 80) The sequence of the sense strand is shown in SEQ ID NO:81, and the sequence of the antisense strand is shown in SEQ ID NO:180; 81) The sequence of the sense strand is shown in SEQ ID NO:82, and the sequence of the antisense strand is shown in SEQ ID NO:181; 82) the sequence of the sense strand is shown in SEQ ID NO:83, and the sequence of the antisense strand is shown in SEQ ID NO:182; 83) the sequence of the sense strand is shown in SEQ ID NO:84, and the sequence of the antisense strand is shown in SEQ ID NO:183; 84) the sequence of the sense strand is shown in SEQ ID NO:85, and the sequence of the antisense strand is shown in SEQ ID NO:184; 85) the sequence of the sense strand is shown in SEQ ID NO:86, and the sequence of the antisense strand is shown in SEQ ID NO:185; 86) The sequence of the sense strand is shown in SEQ ID NO: 87, and the sequence of the antisense strand is shown in SEQ ID NO: 186; 87) The sequence of the sense strand is shown in SEQ ID NO:88, and the sequence of the antisense strand is shown in SEQ ID NO:187; 88) The sequence of the sense strand is shown in SEQ ID NO:89, and the sequence of the antisense strand is shown in SEQ ID NO:188; 89) The sequence of the sense strand is shown in SEQ ID NO:90, and the sequence of the antisense strand is shown in SEQ ID NO:189; 90) The sequence of the sense strand is shown in SEQ ID NO:91, and the sequence of the antisense strand is shown in SEQ ID NO:190; 91) The sequence of the sense strand is shown in SEQ ID NO:92, and the sequence of the antisense strand is shown in SEQ ID NO:191; 92) the sequence of the sense strand is shown in SEQ ID NO:93, and the sequence of the antisense strand is shown in SEQ ID NO:192; 93) the sequence of the sense strand is shown in SEQ ID NO:94, and the sequence of the antisense strand is shown in SEQ ID NO:193; 94) the sequence of the sense strand is shown in SEQ ID NO:95, and the sequence of the antisense strand is shown in SEQ ID NO:194; 95) the sequence of the sense strand is shown in SEQ ID NO:96, and the sequence of the antisense strand is shown in SEQ ID NO:195; 96) the sequence of the sense strand is shown in SEQ ID NO: 97, and the sequence of the antisense strand is shown in SEQ ID NO: 196; 97) The sequence of the sense strand is shown in SEQ ID NO: 98, and the sequence of the antisense strand is shown in SEQ ID NO: 197; 98) the sequence of the sense strand is shown in SEQ ID NO:99, and the sequence of the antisense strand is shown in SEQ ID NO:198; or 99) The sequence of the sense strand is shown in SEQ ID NO: 100, and the sequence of the antisense strand is shown in SEQ ID NO:

199.

5. The siRNA according to any one of claims 1 to 4, wherein Each nucleotide in the nucleotide sequence I and the nucleotide sequence II is a modified nucleotide, and the modified nucleotide is a fluorinated modified nucleotide or a non-fluorinated modified nucleotide; Preferably, the fluorinated modified nucleotide refers to a nucleotide in which the hydroxyl group at the 2'-position of the ribose group of the nucleotide is replaced by fluorine, and has a structure as shown in the following formula (1); the non-fluorinated modified nucleotide refers to a nucleotide or nucleotide analog in which the hydroxyl group at the 2'-position of the ribose group of the nucleotide is replaced by a non-fluorinated group; preferably, the nucleotide or nucleotide analog formed by the non-fluorinated group substitution is a 2'-alkoxy-modified nucleotide or nucleotide analog, a 2'-substituted alkoxy-modified nucleotide or nucleotide analog, a 2'-alkyl-modified nucleotide or nucleotide analog, a 2'-substituted alkyl-modified nucleotide or nucleotide analog, a 2'-amino-modified nucleotide or nucleotide analog, a 2'-substituted amino-modified nucleotide or nucleotide analog, or a 2'-deoxynucleotide; more preferably, the non-fluorinated modified nucleotide has a structure as shown in any one of the following formulas (2) to (12): In formula (1) to formula (12), Base represents a base; In formula (9) and formula (10), R is selected from H, OH or alkoxy (O-alkyl); In formula (11) and formula (12), R is selected from H, OH, F or the non-fluorine group.

6. The siRNA according to claim 5, wherein From the 5' end to the 3' end, one or more nucleotides in positions 7, 9, 10, and 11 of the nucleotide sequence I are fluorinated modified nucleotides; and from the 5' end to the 3' end, one or more nucleotides in positions 2, 6, 8, 9, 14, and 16 of the nucleotide sequence II are fluorinated modified nucleotides.

7. The siRNA according to claim 5 or 6, wherein At least a portion of the phosphate groups in the phosphate-sugar backbone of at least one single strand in the sense strand and the antisense strand of the siRNA is a phosphate group having a modified group, wherein the phosphate group having a modified group is a thiophosphate group formed by replacing at least one oxygen atom in the phosphodiester bond in the phosphate group with a sulfur atom; Preferably, the phosphate group having a modified group is a thiophosphate group having a structure as shown in formula (13): Preferably, the phosphorothioate linkage is present in at least one of the following positions: between the 1st and 2nd nucleotides of the sense strand and / or the antisense strand; between the 2nd and 3rd nucleotides of the sense strand and / or the antisense strand; between the 19th and 20th nucleotides of the antisense strand; between the 20th and 21st nucleotides of the antisense strand; between the 21st and 22nd nucleotides of the antisense strand; between the 22nd and 23rd nucleotides of the antisense strand; or any combination thereof; Preferably, the 5' terminal nucleotide of the antisense strand of the siRNA is a 5'-phosphate nucleotide or a 5'-phosphate analog modified nucleotide, as shown in formula (14), formula (15) and formula (16):

8. An siRNA conjugate comprising the siRNA according to any one of claims 5 to 7 and a conjugated group conjugated to the siRNA, wherein: The conjugated group is L96, and the structure is shown below: Preferably, the siRNA conjugate comprises a sense strand and an antisense strand, which is: 1) The sequence of the sense strand is shown in SEQ ID NO: 200, and the sequence of the antisense strand is shown in SEQ ID NO: 299; 2) the sequence of the sense strand is shown in SEQ ID NO: 201, and the sequence of the antisense strand is shown in SEQ ID NO: 300; 3) the sequence of the sense strand is shown in SEQ ID NO: 202, and the sequence of the antisense strand is shown in SEQ ID NO: 301; 4) the sequence of the sense strand is shown in SEQ ID NO: 203, and the sequence of the antisense strand is shown in SEQ ID NO: 302; 5) the sequence of the sense strand is shown in SEQ ID NO: 204, and the sequence of the antisense strand is shown in SEQ ID NO: 303; 6) the sequence of the sense strand is shown in SEQ ID NO: 205, and the sequence of the antisense strand is shown in SEQ ID NO: 304; 7) the sequence of the sense strand is shown in SEQ ID NO: 206, and the sequence of the antisense strand is shown in SEQ ID NO: 305; 8) the sequence of the sense strand is shown in SEQ ID NO: 207, and the sequence of the antisense strand is shown in SEQ ID NO: 306; 9) the sequence of the sense strand is shown in SEQ ID NO: 208, and the sequence of the antisense strand is shown in SEQ ID NO: 307; 10) the sequence of the sense strand is shown in SEQ ID NO: 209, and the sequence of the antisense strand is shown in SEQ ID NO: 308; 11) The sequence of the sense strand is shown in SEQ ID NO: 210, and the sequence of the antisense strand is shown in SEQ ID NO: 309; 12) the sequence of the sense strand is shown in SEQ ID NO: 211, and the sequence of the antisense strand is shown in SEQ ID NO: 310; 13) the sequence of the sense strand is shown in SEQ ID NO: 212, and the sequence of the antisense strand is shown in SEQ ID NO: 311; 14) the sequence of the sense strand is shown in SEQ ID NO: 213, and the sequence of the antisense strand is shown in SEQ ID NO: 312; 15) the sequence of the sense strand is shown in SEQ ID NO: 214, and the sequence of the antisense strand is shown in SEQ ID NO: 313; 16) the sequence of the sense strand is shown in SEQ ID NO: 215, and the sequence of the antisense strand is shown in SEQ ID NO: 314; 17) the sequence of the sense strand is shown in SEQ ID NO: 216, and the sequence of the antisense strand is shown in SEQ ID NO: 315; 18) the sequence of the sense strand is shown in SEQ ID NO: 217, and the sequence of the antisense strand is shown in SEQ ID NO: 316; 19) The sequence of the sense strand is shown in SEQ ID NO: 218, and the sequence of the antisense strand is shown in SEQ ID NO: 317; 20) the sequence of the sense strand is shown in SEQ ID NO: 219, and the sequence of the antisense strand is shown in SEQ ID NO: 318; 21) the sequence of the sense strand is shown in SEQ ID NO: 220, and the sequence of the antisense strand is shown in SEQ ID NO: 319; 22) the sequence of the sense strand is shown in SEQ ID NO: 221, and the sequence of the antisense strand is shown in SEQ ID NO: 320; 23) the sequence of the sense strand is shown in SEQ ID NO: 222, and the sequence of the antisense strand is shown in SEQ ID NO: 321; 24) the sequence of the sense strand is shown in SEQ ID NO: 223, and the sequence of the antisense strand is shown in SEQ ID NO: 322; 25) the sequence of the sense strand is shown in SEQ ID NO: 224, and the sequence of the antisense strand is shown in SEQ ID NO: 323; 26) the sequence of the sense strand is shown in SEQ ID NO: 225, and the sequence of the antisense strand is shown in SEQ ID NO: 324; 27) the sequence of the sense strand is shown in SEQ ID NO: 226, and the sequence of the antisense strand is shown in SEQ ID NO: 325; 28) the sequence of the sense strand is shown in SEQ ID NO: 227, and the sequence of the antisense strand is shown in SEQ ID NO: 326; 29) the sequence of the sense strand is shown in SEQ ID NO: 228, and the sequence of the antisense strand is shown in SEQ ID NO: 327; 30) the sequence of the sense strand is shown in SEQ ID NO: 229, and the sequence of the antisense strand is shown in SEQ ID NO: 328; 31) the sequence of the sense strand is shown in SEQ ID NO: 230, and the sequence of the antisense strand is shown in SEQ ID NO: 329; 32) the sequence of the sense strand is shown in SEQ ID NO: 231, and the sequence of the antisense strand is shown in SEQ ID NO: 330; 33) the sequence of the sense strand is shown in SEQ ID NO: 232, and the sequence of the antisense strand is shown in SEQ ID NO: 331; 34) the sequence of the sense strand is shown in SEQ ID NO: 233, and the sequence of the antisense strand is shown in SEQ ID NO: 332; 35) the sequence of the sense strand is shown in SEQ ID NO: 234, and the sequence of the antisense strand is shown in SEQ ID NO: 333; 36) the sequence of the sense strand is shown in SEQ ID NO: 235, and the sequence of the antisense strand is shown in SEQ ID NO: 334; 37) the sequence of the sense strand is shown in SEQ ID NO: 236, and the sequence of the antisense strand is shown in SEQ ID NO: 335; 38) the sequence of the sense strand is shown in SEQ ID NO: 237, and the sequence of the antisense strand is shown in SEQ ID NO: 336; 39) the sequence of the sense strand is shown in SEQ ID NO: 238, and the sequence of the antisense strand is shown in SEQ ID NO: 337; 40) the sequence of the sense strand is shown in SEQ ID NO: 239, and the sequence of the antisense strand is shown in SEQ ID NO: 338; 41) The sequence of the sense strand is shown in SEQ ID NO: 240, and the sequence of the antisense strand is shown in SEQ ID NO: 339; 42) the sequence of the sense strand is shown in SEQ ID NO: 241, and the sequence of the antisense strand is shown in SEQ ID NO: 340; 43) the sequence of the sense strand is shown in SEQ ID NO: 242, and the sequence of the antisense strand is shown in SEQ ID NO: 341; 44) the sequence of the sense strand is shown in SEQ ID NO: 243, and the sequence of the antisense strand is shown in SEQ ID NO: 342; 45) the sequence of the sense strand is shown in SEQ ID NO: 244, and the sequence of the antisense strand is shown in SEQ ID NO: 343; 46) the sequence of the sense strand is shown in SEQ ID NO: 245, and the sequence of the antisense strand is shown in SEQ ID NO: 344; 47) the sequence of the sense strand is shown in SEQ ID NO: 246, and the sequence of the antisense strand is shown in SEQ ID NO: 345; 48) The sequence of the sense strand is shown in SEQ ID NO: 247, and the sequence of the antisense strand is shown in SEQ ID NO: 346; 49) The sequence of the sense strand is shown in SEQ ID NO: 248, and the sequence of the antisense strand is shown in SEQ ID NO: 347; 50) the sequence of the sense strand is shown in SEQ ID NO: 249, and the sequence of the antisense strand is shown in SEQ ID NO: 348; 51) The sequence of the sense strand is shown in SEQ ID NO: 250, and the sequence of the antisense strand is shown in SEQ ID NO: 349; 52) the sequence of the sense strand is shown in SEQ ID NO: 251, and the sequence of the antisense strand is shown in SEQ ID NO: 350; 53) the sequence of the sense strand is shown in SEQ ID NO: 252, and the sequence of the antisense strand is shown in SEQ ID NO: 351; 54) the sequence of the sense strand is shown in SEQ ID NO: 253, and the sequence of the antisense strand is shown in SEQ ID NO: 352; 55) the sequence of the sense strand is shown in SEQ ID NO: 254, and the sequence of the antisense strand is shown in SEQ ID NO: 353; 56) The sequence of the sense strand is shown in SEQ ID NO: 255, and the sequence of the antisense strand is shown in SEQ ID NO: 354; 57) The sequence of the sense strand is shown in SEQ ID NO: 256, and the sequence of the antisense strand is shown in SEQ ID NO: 355; 58) The sequence of the sense strand is shown in SEQ ID NO: 257, and the sequence of the antisense strand is shown in SEQ ID NO: 356; 59) The sequence of the sense strand is shown in SEQ ID NO: 258, and the sequence of the antisense strand is shown in SEQ ID NO: 357; 60) The sequence of the sense strand is shown in SEQ ID NO: 259, and the sequence of the antisense strand is shown in SEQ ID NO: 358; 61) The sequence of the sense strand is shown in SEQ ID NO: 260, and the sequence of the antisense strand is shown in SEQ ID NO: 359; 62) The sequence of the sense strand is shown in SEQ ID NO: 261, and the sequence of the antisense strand is shown in SEQ ID NO: 360; 63) The sequence of the sense strand is shown in SEQ ID NO: 262, and the sequence of the antisense strand is shown in SEQ ID NO: 361; 64) The sequence of the sense strand is shown in SEQ ID NO: 263, and the sequence of the antisense strand is shown in SEQ ID NO: 362; 65) The sequence of the sense strand is shown in SEQ ID NO: 264, and the sequence of the antisense strand is shown in SEQ ID NO: 363; 66) The sequence of the sense strand is shown in SEQ ID NO: 265, and the sequence of the antisense strand is shown in SEQ ID NO: 364; 67) The sequence of the sense strand is shown in SEQ ID NO: 266, and the sequence of the antisense strand is shown in SEQ ID NO: 365; 68) The sequence of the sense strand is shown in SEQ ID NO: 267, and the sequence of the antisense strand is shown in SEQ ID NO: 366; 69) The sequence of the sense strand is shown in SEQ ID NO: 268, and the sequence of the antisense strand is shown in SEQ ID NO: 367; 70) The sequence of the sense strand is shown in SEQ ID NO: 269, and the sequence of the antisense strand is shown in SEQ ID NO: 368; 71) The sequence of the sense strand is shown in SEQ ID NO: 270, and the sequence of the antisense strand is shown in SEQ ID NO: 369; 72) The sequence of the sense strand is shown in SEQ ID NO: 271, and the sequence of the antisense strand is shown in SEQ ID NO: 370; 73) The sequence of the sense strand is shown in SEQ ID NO: 272, and the sequence of the antisense strand is shown in SEQ ID NO: 371; 74) the sequence of the sense strand is shown in SEQ ID NO: 273, and the sequence of the antisense strand is shown in SEQ ID NO: 372; 75) The sequence of the sense strand is shown in SEQ ID NO: 274, and the sequence of the antisense strand is shown in SEQ ID NO: 373; 76) The sequence of the sense strand is shown in SEQ ID NO: 275, and the sequence of the antisense strand is shown in SEQ ID NO: 374; 77) The sequence of the sense strand is shown in SEQ ID NO: 276, and the sequence of the antisense strand is shown in SEQ ID NO: 375; 78) The sequence of the sense strand is shown in SEQ ID NO: 277, and the sequence of the antisense strand is shown in SEQ ID NO: 376; 79) The sequence of the sense strand is shown in SEQ ID NO: 278, and the sequence of the antisense strand is shown in SEQ ID NO: 377; 80) The sequence of the sense strand is shown in SEQ ID NO: 279, and the sequence of the antisense strand is shown in SEQ ID NO: 378; 81) The sequence of the sense strand is shown in SEQ ID NO: 280, and the sequence of the antisense strand is shown in SEQ ID NO: 379; 82) the sequence of the sense strand is shown in SEQ ID NO: 281, and the sequence of the antisense strand is shown in SEQ ID NO: 380; 83) The sequence of the sense strand is shown in SEQ ID NO: 282, and the sequence of the antisense strand is shown in SEQ ID NO: 381; 84) the sequence of the sense strand is shown in SEQ ID NO: 283, and the sequence of the antisense strand is shown in SEQ ID NO: 382; 85) The sequence of the sense strand is shown in SEQ ID NO: 284, and the sequence of the antisense strand is shown in SEQ ID NO: 383; 86) The sequence of the sense strand is shown in SEQ ID NO: 285, and the sequence of the antisense strand is shown in SEQ ID NO: 384; 87) The sequence of the sense strand is shown in SEQ ID NO: 286, and the sequence of the antisense strand is shown in SEQ ID NO: 385; 88) The sequence of the sense strand is shown in SEQ ID NO: 287, and the sequence of the antisense strand is shown in SEQ ID NO: 386; 89) The sequence of the sense strand is shown in SEQ ID NO: 288, and the sequence of the antisense strand is shown in SEQ ID NO: 387; 90) The sequence of the sense strand is shown in SEQ ID NO: 289, and the sequence of the antisense strand is shown in SEQ ID NO: 388; 91) The sequence of the sense strand is shown in SEQ ID NO: 290, and the sequence of the antisense strand is shown in SEQ ID NO: 389; 92) the sequence of the sense strand is shown in SEQ ID NO: 291, and the sequence of the antisense strand is shown in SEQ ID NO: 390; 93) the sequence of the sense strand is shown in SEQ ID NO: 292, and the sequence of the antisense strand is shown in SEQ ID NO: 391; 94) the sequence of the sense strand is shown in SEQ ID NO: 293, and the sequence of the antisense strand is shown in SEQ ID NO: 392; 95) the sequence of the sense strand is shown in SEQ ID NO: 294, and the sequence of the antisense strand is shown in SEQ ID NO: 393; 96) The sequence of the sense strand is shown in SEQ ID NO: 295, and the sequence of the antisense strand is shown in SEQ ID NO: 394; 97) The sequence of the sense strand is shown in SEQ ID NO: 296, and the sequence of the antisense strand is shown in SEQ ID NO: 395; 98) the sequence of the sense strand is shown in SEQ ID NO: 297, and the sequence of the antisense strand is shown in SEQ ID NO: 396; or 99) The sequence of the sense strand is shown in SEQ ID NO:298, and the sequence of the antisense strand is shown in SEQ ID NO:

397.

9. A composition comprising the siRNA according to any one of claims 1 to 7 or the siRNA conjugate according to claim 8; Preferably, the composition is a pharmaceutical composition, further comprising a pharmaceutically acceptable carrier or excipient; Preferably, the carrier or excipient includes but is not limited to water for injection, sodium hydroxide, sodium dihydrogen phosphate monohydrate, sodium dihydrogen phosphate dihydrate, phosphoric acid, sodium chloride, potassium chloride, hydrochloric acid, anhydrous potassium dihydrogen phosphate, anhydrous disodium hydrogen phosphate, PEG2000, PEG6000, cholesterol, distearoylphosphatidylcholine, 1,2-dimyristyl glyceride, and dimethyl adipate.

10. Use of the siRNA according to any one of claims 1 to 7, the siRNA conjugate according to claim 8, or the pharmaceutical composition according to claim 9 in the preparation of a drug for preventing and / or treating a disease associated with PLN expression; Preferably, the diseases associated with PLN expression include one or more of heart failure, cardiomyopathy, and arrhythmia; Preferably, the heart failure includes one or more of acute heart failure, chronic heart failure, systolic heart failure, diastolic heart failure, left heart failure, right heart failure, and total heart failure; Preferably, the cardiomyopathy is a hereditary cardiomyopathy, including one or more of cardiomyopathy caused by p.Arg14del, Arg9Cys (R9C) and / or Arg25Cys (R25C) gene mutations; More preferably, the cardiomyopathy is dilated cardiomyopathy, including one or more of dilated cardiomyopathy caused by TTN, LMNA, RBM20, SCN5A, MYH7, TNNT2 and / or TPM1 mutations; Preferably, the arrhythmia comprises ventricular tachycardia and / or ventricular fibrillation.

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