Sirna for inhibiting gene expression of LPA, composition containing same, and use thereof

Abstract

The present invention relates to an siRNA for inhibiting the gene expression of LPA, a composition containing same, and a use thereof. The siRNA comprises a sense strand and an antisense strand, wherein the antisense strand comprises a sequence complementary to an LPA mRNA sequence, and the sequence of the antisense strand includes a sequence of any antisense strand in Table 1.

Description

siRNA for inhibiting gene expression of LPA, composition containing the same and use thereof Technical Field

[0001] The present application relates to the field of biomedicine, and in particular to siRNA for inhibiting the gene expression of LPA, a composition containing the siRNA, and uses thereof. Background Art

[0002] LPA is the name of the gene encoding apolipoprotein (a) (apo(a)). It is primarily expressed in the liver, with expression restricted to humans and non-primates. Apolipoprotein (a) is attached to apo(B)-100 via a disulfide bond, forming a lipid core that forms the lipoprotein (a) (Lp(a)) particle. Lp(a) particles are specialized, large cholesterol-rich lipoproteins with a surface coated with cholesterol and phospholipids and embedded with the hydrophilic apolipoprotein components apolipoprotein (a) and apo(B)-100. Lp(a) can enter and deposit on blood vessel walls, promoting atherosclerosis. Lp(a) is structurally homologous to plasminogen (PLG) and can compete with plasminogen for binding sites on fibrin, thereby inhibiting fibrinogen hydrolysis and promoting thrombosis. Therefore, Lp(a) is closely associated with atherosclerosis and thrombosis. Studies have shown that Lp(a) levels in the blood are an independent risk factor for cardiovascular disease, stroke, and atherosclerotic stenosis.

[0003] Analysis of Lp(a) levels in multiple studies has suggested that high Lp(a) levels are an independent risk factor for cardiovascular disease, stroke, and other related conditions, including atherosclerotic stenosis. Furthermore, genome-wide association studies have also implicated LPA as a genetic risk factor for diseases such as atherosclerotic stenosis.

[0004] Therefore, there is a need for methods for effectively treating, preventing, and reducing the risk of developing conditions such as and associated with stroke, atherosclerosis, thrombosis, and cardiovascular diseases such as coronary heart disease, aortic stenosis, and other as yet unidentified related conditions, pathologies, or syndromes. The present invention addresses this unmet medical need.

[0005] Summary of the Invention

[0006] The purpose of this article is to provide siRNA (referred to herein as LPA siRNA) or a pharmaceutically acceptable salt thereof for inhibiting the expression of the LPA gene. The siRNA described herein, after delivery to cells expressing the LPA gene, inhibits and / or knocks down LPA expression in vitro and / or in vivo through the biological process of RNA interference (RNAi).

[0007] Another object of the present invention is to provide a pharmaceutical composition containing the siRNA or a pharmaceutically acceptable salt thereof.

[0008] Another object of the present invention is to provide use of the siRNA or a pharmaceutically acceptable salt thereof or the pharmaceutical composition in preventing or treating diseases or conditions related to LPA expression.

[0009] To achieve the above object, the present invention provides an siRNA or a pharmaceutically acceptable salt thereof, comprising a sense strand and an antisense strand, wherein the antisense strand comprises a sequence complementary to the LPA mRNA sequence, and the sequence of the antisense strand comprises any one of the antisense strand sequences in Table 1:

[0010] Table 1

[0011] In some embodiments, the antisense strand is 19-26 nucleobases long.

[0012] In some embodiments, the antisense strand is 19-21 nucleobases long.

[0013] In some embodiments, the sense strand comprises a sequence that is substantially complementary to the antisense strand and forms a duplex complementary region of 15 to 26 consecutive base pairs.

[0014] In some embodiments, the length of the duplex complementary region is 18 to 23 consecutive base pairs.

[0015] In some embodiments, the length of the duplex complementary region is 19 to 21 consecutive base pairs.

[0016] In some embodiments, the sense strand is 19-26 nucleobases long.

[0017] In some embodiments, the sense strand is 19-21 nucleobases long.

[0018] In some embodiments, the sense strand sequence comprises any sense strand sequence in Table 1.

[0019] In some embodiments, the antisense strand comprises any one of SEQ ID NOs. 128-150, 253-254.

[0020] In some embodiments, the sense strand comprises any one of SEQ ID NOs. 1-23, 126-127.

[0021] In some embodiments, the siRNA comprises any one of the siRNAs listed in Table 1.

[0022] In some embodiments, in the siRNA:

[0023] The antisense strand comprises SEQ ID NO. 128, and the sense strand comprises SEQ ID NO. 1;

[0024] The antisense strand comprises SEQ ID NO. 129, and the sense strand comprises SEQ ID NO. 2;

[0025] The antisense strand comprises SEQ ID NO. 130, and the sense strand comprises SEQ ID NO. 3;

[0026] The antisense strand comprises SEQ ID NO. 131, and the sense strand comprises SEQ ID NO. 4;

[0027] The antisense strand comprises SEQ ID NO. 132, and the sense strand comprises SEQ ID NO. 5;

[0028] The antisense strand comprises SEQ ID NO. 133, and the sense strand comprises SEQ ID NO. 6;

[0029] The antisense strand comprises SEQ ID NO. 134, and the sense strand comprises SEQ ID NO. 7;

[0030] The antisense strand comprises SEQ ID NO. 135, and the sense strand comprises SEQ ID NO. 8;

[0031] The antisense strand comprises SEQ ID NO. 136, and the sense strand comprises SEQ ID NO. 9;

[0032] The antisense strand comprises SEQ ID NO. 137, and the sense strand comprises SEQ ID NO. 10;

[0033] The antisense strand comprises SEQ ID NO. 138, and the sense strand comprises SEQ ID NO. 11;

[0034] The antisense strand comprises SEQ ID NO. 139, and the sense strand comprises SEQ ID NO. 12;

[0035] The antisense strand comprises SEQ ID NO. 140, and the sense strand comprises SEQ ID NO. 13;

[0036] The antisense strand comprises SEQ ID NO. 141, and the sense strand comprises SEQ ID NO. 14;

[0037] The antisense strand comprises SEQ ID NO. 142, and the sense strand comprises SEQ ID NO. 15;

[0038] The antisense strand comprises SEQ ID NO. 143, and the sense strand comprises SEQ ID NO. 16;

[0039] The antisense strand comprises SEQ ID NO. 144, and the sense strand comprises SEQ ID NO. 17;

[0040] The antisense strand comprises SEQ ID NO. 145, and the sense strand comprises SEQ ID NO. 18;

[0041] The antisense strand comprises SEQ ID NO. 146, and the sense strand comprises SEQ ID NO. 19;

[0042] The antisense strand comprises SEQ ID NO. 147, and the sense strand comprises SEQ ID NO. 20;

[0043] The antisense strand comprises SEQ ID NO. 148, and the sense strand comprises SEQ ID NO. 21;

[0044] The antisense strand comprises SEQ ID NO. 149, and the sense strand comprises SEQ ID NO. 22;

[0045] The antisense strand comprises SEQ ID NO. 150, and the sense strand comprises SEQ ID NO. 23;

[0046] The antisense strand includes SEQ ID NO.253, and the sense strand includes SEQ ID NO.126 or

[0047] The antisense strand comprises SEQ ID NO. 254, and the sense strand comprises SEQ ID NO. 127.

[0048] In some embodiments, the antisense strand or sense strand contains 1, 2, 3, 4, 5, or 6 extensions.

[0049] In some embodiments, the antisense strand or sense strand is extended by dT.

[0050] In some embodiments, the antisense strand or the sense strand contains two dT stretches.

[0051] In some embodiments, the 3' end of the antisense strand contains two extended dTs, and the sense strand does not contain dTs.

[0052] In some embodiments, the antisense strand comprises the sequence of any one of the antisense strands in Table 2:

[0053] Table 2

[0054] In some embodiments, the antisense strand comprises any one of SEQ ID NOs. 380-402, 544-546.

[0055] In some embodiments, the sense strand comprises the sequence of any sense strand in Table 2.

[0056] In some embodiments, the sense strand comprises any one of SEQ ID NOs. 255-277, 541-543.

[0057] In some embodiments, the siRNA comprises any one of the siRNAs listed in Table 2.

[0058] In some embodiments, in the siRNA:

[0059] The antisense strand comprises SEQ ID NO. 380, and the sense strand comprises SEQ ID NO. 255;

[0060] The antisense strand comprises SEQ ID NO. 381, and the sense strand comprises SEQ ID NO. 256;

[0061] The antisense strand comprises SEQ ID NO. 382, ​​and the sense strand comprises SEQ ID NO. 257;

[0062] The antisense strand comprises SEQ ID NO. 383, and the sense strand comprises SEQ ID NO. 258;

[0063] The antisense strand comprises SEQ ID NO. 384, and the sense strand comprises SEQ ID NO. 259;

[0064] The antisense strand comprises SEQ ID NO. 385, and the sense strand comprises SEQ ID NO. 260;

[0065] The antisense strand comprises SEQ ID NO. 386, and the sense strand comprises SEQ ID NO. 261;

[0066] The antisense strand comprises SEQ ID NO. 387, and the sense strand comprises SEQ ID NO. 262;

[0067] The antisense strand comprises SEQ ID NO. 388, and the sense strand comprises SEQ ID NO. 263;

[0068] The antisense strand comprises SEQ ID NO. 389, and the sense strand comprises SEQ ID NO. 264;

[0069] The antisense strand comprises SEQ ID NO. 390, and the sense strand comprises SEQ ID NO. 265;

[0070] The antisense strand comprises SEQ ID NO. 391, and the sense strand comprises SEQ ID NO. 266;

[0071] The antisense strand comprises SEQ ID NO. 392, and the sense strand comprises SEQ ID NO. 267;

[0072] The antisense strand comprises SEQ ID NO. 393, and the sense strand comprises SEQ ID NO. 268;

[0073] The antisense strand comprises SEQ ID NO. 394, and the sense strand comprises SEQ ID NO. 269;

[0074] The antisense strand comprises SEQ ID NO. 395, and the sense strand comprises SEQ ID NO. 270;

[0075] The antisense strand comprises SEQ ID NO. 396, and the sense strand comprises SEQ ID NO. 271;

[0076] The antisense strand comprises SEQ ID NO. 397, and the sense strand comprises SEQ ID NO. 272;

[0077] The antisense strand comprises SEQ ID NO. 398, and the sense strand comprises SEQ ID NO. 273;

[0078] The antisense strand comprises SEQ ID NO. 399, and the sense strand comprises SEQ ID NO. 274;

[0079] The antisense strand comprises SEQ ID NO. 400, and the sense strand comprises SEQ ID NO. 275;

[0080] The antisense strand comprises SEQ ID NO. 401, and the sense strand comprises SEQ ID NO. 276;

[0081] The antisense strand comprises SEQ ID NO. 402, and the sense strand comprises SEQ ID NO. 277;

[0082] The antisense strand comprises SEQ ID NO. 541, and the sense strand comprises SEQ ID NO. 544;

[0083] The antisense strand comprises SEQ ID NO. 542, and the sense strand comprises SEQ ID NO. 545; or

[0084] The antisense strand comprises SEQ ID NO. 543, and the sense strand comprises SEQ ID NO. 546.

[0085] In some embodiments, the sense strand and / or antisense strand independently comprises one or more modified nucleotides.

[0086] In some embodiments, the modified nucleotide is a 2'-modified nucleotide.

[0087] In some embodiments, the modified nucleotides include: 2'-O-methyl modified nucleotides (i.e., ribonucleotides in which the 2'-OH on the ribose is replaced by 2'-OMe, represented herein as mN, where N represents a nucleotide), 2'-O-methoxyethyl modified nucleotides, 2'-fluoro modified nucleotides (i.e., ribonucleotides in which the 2'-OH on the ribose is replaced by 2'-F, represented herein as fN, where N represents a ribonucleotide, and also represented herein as 2'-deoxy-2'-fluoro modified nucleotides), 2'-deoxy modified nucleotides, 2'-O-alkenyl modified nucleotides, locked nucleotides, GNA, LNA, abasic nucleotides, deoxythymidine, inverted deoxythymidine, deoxyadenosine, inverted deoxyadenosine, 2'-amino-modified nucleotides, 2'-alkyl-modified nucleotides, morpholino nucleotides and nucleotides including non-natural bases, 5'-(E)-vinyl phosphate modified nucleotides (represented herein as VP), or combinations thereof.

[0088] In some embodiments, the nucleotides are all modified nucleotides.

[0089] In some embodiments, the modified nucleotides include 2'-O-methyl modified nucleotides, 2'-fluoro modified nucleotides, deoxythymidine and 5'-(E)-vinyl phosphate modified nucleotides, or a combination thereof.

[0090] In some embodiments, the sense strand, the antisense strand, or both the sense strand and the antisense strand comprise one or more phosphorodiester monothioate or phosphorodiester internucleoside linkages.

[0091] In some embodiments, the antisense strand comprises two consecutive monothioate phosphorodiester internucleoside linkages between the 3' and 5' terminal nucleotides.

[0092] In some embodiments, the sense strand comprises two consecutive monothioate phosphorodiester internucleoside linkages between the 3' and 5' terminal nucleotides.

[0093] In some embodiments, the 5' terminal nucleotides of the sense strand include two consecutive monothioate phosphorodiester internucleoside linkages, and the 3' terminal nucleotides of the sense strand include one monothioate phosphorodiester internucleoside linkage.

[0094] In some embodiments, the sense strand comprises two consecutive monothioate phosphorodiester nucleoside linkages between its 5' terminal nucleotides, the sense strand comprises one monothioate phosphorodiester nucleoside linkage between its 3' terminal nucleotides; and the antisense strand comprises two consecutive monothioate phosphorodiester nucleoside linkages between its 3' and 5' terminal nucleotides.

[0095] In some embodiments:

[0096] 1) The sense strand comprises 21 nucleotides, the antisense strand comprises 23 nucleotides, the duplex complementary region is 21 base pairs in length, the 9th, 10th, and 11th nucleotides from the 5' end of the sense strand are all 2'-fluorinated nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl modified nucleotides; the 8th, 10th, 18th, and 22nd nucleotides from the 3' end of the antisense strand are all 2'-fluorinated nucleotides, and the remaining nucleotides of the antisense strand are all 2'-O-methyl modified nucleotides; the 1st to 3rd nucleotides from the 5' end of the sense strand, the 1st to 3rd nucleotides from the 3' end of the sense strand, the 1st to 3rd nucleotides from the 5' end of the antisense strand, and the 1st to 3rd nucleotides from the 3' end of the antisense strand are all linked by monothiophosphate diester bonds, and the remaining nucleotides of the sense and antisense strands are linked by phosphodiester bonds;

[0097] 2) The sense strand comprises 19 nucleotides, the antisense strand comprises 21 nucleotides, the length of the duplex complementary region is 19 base pairs, the 7th, 8th and 9th nucleotides from the 5' end of the sense strand are all 2'-fluorinated nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl modified nucleotides; the 1st and 2nd positions of the antisense strand from the 3' end are two dTs, and the 6th, 8th, 14th, 16th and 20th nucleotides from the 3' end of the antisense strand are all 2' -fluorinated nucleotides, and the remaining nucleotides of the antisense strand are all 2'-O-methyl modified nucleotides; the first to third nucleotides from the 5' end of the sense strand, the first to third nucleotides from the 3' end of the sense strand, the first to third nucleotides from the 5' end of the antisense strand, and the first to third nucleotides from the 3' end of the antisense strand are all linked by monothioate phosphorodiester bonds, and the remaining nucleotides of the sense and antisense strands are linked by phosphodiester bonds;

[0098] 3) The sense strand comprises 19 nucleotides, the antisense strand comprises 21 nucleotides, the length of the duplex complementary region is 19 base pairs, the 7th, 8th and 9th nucleotides from the 5' end of the sense strand are all 2'-fluorinated nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl modified nucleotides; the 1st and 2nd positions of the antisense strand from the 3' end are two dTs, and the 4th, 6th, 8th, 16th and 20th nucleotides from the 3' end of the antisense strand are all 2'- Fluorine-modified nucleotides, the remaining nucleotides of the antisense strand are all 2'-O-methyl modified nucleotides; the first to third nucleotides from the 5' end of the sense strand, the first to third nucleotides from the 3' end of the sense strand, the first to third nucleotides from the 5' end of the antisense strand, and the first to third nucleotides from the 3' end of the antisense strand are all connected by monothioate phosphorodiester bonds, and the remaining nucleotides of the sense and antisense strands are connected by phosphodiester bonds;

[0099] 4) The sense strand comprises 21 nucleotides, the antisense strand comprises 21 nucleotides, and the length of the duplex complementary region is 21 base pairs. The 9th, 10th, and 11th nucleotides from the 5' end of the sense strand are all 2'-fluorinated nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl modified nucleotides; the 4th, 6th, 8th, 10th, 16th, 18th, and 20th nucleotides from the 3' end of the antisense strand are all 2'-fluorinated nucleotides, and the remaining nucleotides of the antisense strand are modified with 2'-O-methyl modified nucleotides; the 1st to 3rd nucleotides from the 5' end of the sense strand, the 1st to 3rd nucleotides from the 3' end of the sense strand, the 1st to 3rd nucleotides from the 5' end of the antisense strand, and the 1st to 3rd nucleotides from the 3' end of the antisense strand are all connected by monothiophosphate diester bonds, and the remaining nucleotides of the sense and antisense strands are connected by phosphodiester bonds;

[0100] 5) The sense strand comprises 21 nucleotides, the antisense strand comprises 21 nucleotides, the length of the duplex complementary region is 21 base pairs, the 9th, 10th and 11th nucleotides from the 5' end of the sense strand are all 2'-fluorinated nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl modified nucleotides; the 4th, 6th, 8th, 10th, 16th, 18th and 20th nucleotides from the 3' end of the antisense strand are all 2'-fluorinated nucleotides, and the remaining nucleotides of the antisense strand are The amino acid is modified with a 2'-O-methyl modified nucleotide, and the first nucleotide at the 5' end of the antisense strand is modified with VP; the first to third nucleotides from the 5' end of the sense strand, the first to third nucleotides from the 3' end of the sense strand, the first to third nucleotides from the 5' end of the antisense strand, and the first to third nucleotides from the 3' end of the antisense strand are all connected by monothioate phosphorodiester bonds, and the remaining nucleotides of the sense and antisense strands are connected by phosphodiester bonds;

[0101] 6) The sense strand comprises 19 nucleotides, the antisense strand comprises 21 nucleotides, the length of the duplex complementary region is 19 base pairs, the 7th, 8th and 9th nucleotides from the 5' end of the sense strand are all 2'-fluorinated nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl modified nucleotides; the 1st and 2nd positions of the antisense strand from the 3' end are two dTs, the 6th, 8th, 14th, 16th and 20th nucleotides from the 3' end of the antisense strand are all 2'-fluorinated nucleotides, and the remaining nucleotides of the antisense strand are all 2'-O-methyl modified nucleotides. The remaining nucleotides of the sense strand are all 2'-O-methyl modified nucleotides, and the first nucleotide at the 5' end of the antisense strand is modified with VP; the first to third nucleotides from the 5' end of the sense strand, the first to third nucleotides from the 3' end of the sense strand, the first to third nucleotides from the 5' end of the antisense strand, and the first to third nucleotides from the 3' end of the antisense strand are all connected by monothioate phosphorodiester bonds, and the remaining nucleotides of the sense and antisense strands are connected by phosphodiester bonds;

[0102] 7) The sense strand comprises 19 nucleotides, the antisense strand comprises 21 nucleotides, the length of the duplex complementary region is 19 base pairs, the 7th, 8th and 9th nucleotides from the 5' end of the sense strand are all 2'-fluorinated nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl modified nucleotides; the 1st and 2nd positions of the antisense strand from the 3' end are two dTs, and the 6th, 8th, 14th, 16th and 20th nucleotides from the 3' end of the antisense strand are all 2'- Fluorine-modified nucleotides, the remaining nucleotides of the antisense strand are all 2'-O-methyl modified nucleotides; the first to third nucleotides from the 5' end of the sense strand, the first to second nucleotides from the 3' end of the sense strand, the first to third nucleotides from the 5' end of the antisense strand, and the first to third nucleotides from the 3' end of the antisense strand are all linked by monothioate phosphorodiester bonds, and the remaining nucleotides of the sense and antisense strands are linked by phosphodiester bonds; or

[0103] 8) The sense strand comprises 19 nucleotides, the antisense strand comprises 21 nucleotides, the length of the duplex complementary region is 19 base pairs, the 7th, 8th and 9th nucleotides from the 5' end of the sense strand are all 2'-fluorinated nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl modified nucleotides; the 1st and 2nd positions of the antisense strand from the 3' end are two dTs, the 6th, 8th, 14th, 16th and 20th nucleotides from the 3' end of the antisense strand are all 2'-fluorinated nucleotides, and the remaining nucleotides of the antisense strand are all 2'-O-methyl modified nucleotides. The remaining nucleotides of the sense strand are all 2'-O-methyl modified nucleotides, and the first nucleotide at the 5' end of the antisense strand is modified with VP; the first to third nucleotides from the 5' end of the sense strand, the first to second nucleotides from the 3' end of the sense strand, the first to third nucleotides from the 5' end of the antisense strand, and the first to third nucleotides from the 3' end of the antisense strand are all connected by monothioate phosphorodiester bonds, and the remaining nucleotides of the sense and antisense strands are connected by phosphodiester bonds.

[0104] In some embodiments, the antisense strand comprises any one of the antisense strands in Table 3:

[0105] Table 3

[0106] In Table 3, fA, fC, fU, and fG represent 2'-fluoro-modified A, C, U, and G ribonucleoside-3'-phosphates, respectively; mA, mC, mU, and mG represent 2'-O-methyl-modified A, C, U, and G ribonucleoside-3'-phosphates, respectively; * represents a monothioate phosphorodiester bond; dT represents thymidine deoxyribonucleoside-3'-phosphate; and VP represents a ribonucleotide modified with a 5'-(E)-vinyl phosphate group.

[0107] In some embodiments, the antisense strand comprises the antisense strand of any one of SEQ ID NOs. 505, 507, 509, 511, 513, 515, 517, 519, 521, 523, 525, and 527.

[0108] In some embodiments, the sense strand comprises any one of the sense strands in Table 3.

[0109] In some embodiments, the sense strand comprises any one of the sense strands in Table SEQ ID NO. 506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526, and 528.

[0110] In some embodiments, the siRNA comprises any one of the siRNAs listed in Table 3.

[0111] In some embodiments, in the siRNA:

[0112] The antisense strand comprises SEQ ID NO. 505, and the sense strand comprises SEQ ID NO. 506;

[0113] The antisense strand comprises SEQ ID NO. 507, and the sense strand comprises SEQ ID NO. 508;

[0114] The antisense strand comprises SEQ ID NO. 509, and the sense strand comprises SEQ ID NO. 510;

[0115] The antisense strand comprises SEQ ID NO. 511, and the sense strand comprises SEQ ID NO. 512;

[0116] The antisense strand comprises SEQ ID NO. 513, and the sense strand comprises SEQ ID NO. 514;

[0117] The antisense strand comprises SEQ ID NO. 515, and the sense strand comprises SEQ ID NO. 516;

[0118] The antisense strand comprises SEQ ID NO. 517, and the sense strand comprises SEQ ID NO. 518;

[0119] The antisense strand comprises SEQ ID NO. 519, and the sense strand comprises SEQ ID NO. 520;

[0120] The antisense strand comprises SEQ ID NO. 521, and the sense strand comprises SEQ ID NO. 522;

[0121] The antisense strand comprises SEQ ID NO. 523, and the sense strand comprises SEQ ID NO. 524;

[0122] The antisense strand comprises SEQ ID NO. 525, and the sense strand comprises SEQ ID NO. 526; or

[0123] The antisense strand comprises SEQ ID NO.527, and the sense strand comprises SEQ ID NO.528.

[0124] In some embodiments, the siRNA further comprises a targeting group.

[0125] In some embodiments, the targeting group comprises an asialoglycoprotein receptor ligand.

[0126] In some embodiments, the asialoglycoprotein receptor ligand comprises a galactose cluster.

[0127] In some embodiments, the galactose cluster is a monovalent, divalent, trivalent, or tetravalent galactose derivative.

[0128] In some embodiments, the galactose derivative is N-acetylgalactosamine (GalNAc).

[0129] In some embodiments, the galactose cluster is trivalent N-acetylgalactosamine.

[0130] In some embodiments, the galactose cluster is connected to the branching center via a linker, and the branching center is conjugated to the nucleotides of the siRNA via the linker.

[0131] In some embodiments, the targeting group is conjugated to the 5' end of the sense strand.

[0132] In some embodiments, the targeting group is conjugated to the 3' end of the sense strand.

[0133] In some embodiments, the following fragment comprising a targeting group is conjugated to the 5' end or 3' end of the sense strand (ie, the following targeting group replaces the hydroxyl group on the 5' end or 3' end):

[0134] In some embodiments, the fragment comprising the targeting group is conjugated to the 5' end of the sense strand.

[0135] In some embodiments, the fragment comprising the targeting group is conjugated to the 3' end of the sense strand.

[0136] In some embodiments, the sense strand is any sense strand listed in Table 4.

[0137] In some embodiments, the antisense strand is any antisense strand in Table 4.

[0138] In some embodiments, the siRNA is any one of the siRNAs listed in Table 4.

[0139] Table 4

[0140] In Table 4, fA, fC, fU, and fG represent 2'-fluoro-modified A, C, U, and G ribonucleoside-3'-phosphates, respectively; mA, mC, mU, and mG represent 2'-O-methyl-modified A, C, U, and G ribonucleoside-3'-phosphates, respectively; * represents a monothioate phosphorodiester bond. If no * appears between two nucleotides, it means that the two are connected by a normal phosphodiester bond; dT represents thymidine deoxyribonucleoside-3'-phosphate; P represents that the nucleotide adjacent to the right of the P is a 5'-phosphate-modified nucleotide; invdA represents an inverted deoxyribonucleotide (3'-3' linked nucleoside). NAG25-AMG890 is olpasiran, and GalNAC-SLN360 is zerlasiran, wherein NAG25 and [ST23*]3 / C6XLT have corresponding structures; Reference Sequence 1 was modified using the method disclosed in CN117327698A, Reference Sequence 2 was modified using the method disclosed in CN116801886A, NAG25-AMG890 was modified using the method disclosed in US9932586B, and GalNAC-SLN360 was modified using the method disclosed in US11499153B. *L96 indicates:

[0141] In some embodiments, all hydroxyl groups or sulfhydryl groups on the phosphodiester bonds or monothiosulfate diester bonds in the siRNA are present in the form of Na salts. ).

[0142] In another aspect, the present invention provides a pharmaceutical composition comprising the siRNA or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable salt thereof according to the present invention.

[0143] In another aspect, the present invention provides a method for reducing the expression level of LPA in a subject, comprising administering to the subject the siRNA of the present invention or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the present invention.

[0144] In another aspect, the present invention provides use of the siRNA described herein, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described herein, in the preparation of a medicament for preventing or treating a disease associated with LPA expression. In some embodiments, the disease associated with LPA expression is a cardiovascular disease. In some embodiments, the cardiovascular disease includes stroke, atherosclerosis, thrombosis, coronary heart disease, or aortic valve stenosis, as well as any other disease or pathology associated with elevated levels of Lp(a)-containing particles.

[0145] In another aspect, the present invention provides a method for preventing or treating a disease associated with LPA expression, comprising administering to a subject an siRNA described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein. In some embodiments, the disease associated with LPA expression is a cardiovascular disease. In some embodiments, the cardiovascular disease includes stroke, atherosclerosis, thrombosis, coronary heart disease, or aortic stenosis, as well as any other disease or pathology associated with elevated levels of Lp(a)-containing particles.

[0146] In another aspect, the present invention provides an siRNA or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition, for use in preventing or treating a disease associated with LPA expression. In some embodiments, the disease associated with LPA expression is a cardiovascular disease. In some embodiments, the cardiovascular disease includes stroke, atherosclerosis, thrombosis, coronary heart disease, or aortic stenosis, as well as any other disease or pathology associated with elevated levels of Lp(a)-containing particles.

[0147] Definition of terms

[0148] The term siRNA (small interfering RNA) refers to a short double-stranded RNA that mediates sequence-specific and effective inhibition of gene expression (which may also be referred to as gene silencing).

[0149] The term "modified nucleotides" refers to nucleotides other than ribonucleotides (2'-hydroxy nucleotides). In some embodiments, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or 100% of the nucleotides are modified. Modified nucleotides include, but are not limited to, deoxynucleotides, nucleotide mimetics, abasic nucleotides, 2'-modified nucleotides, 3' to 3' linkage (inverted) nucleotides, nucleotides comprising non-natural bases, bridged nucleotides, peptide nucleic acids (PNA), 2',3'-bromonucleotide mimics (unlocked nucleobase analogs), locked nucleotides, 3'-O-methoxy (2' internucleoside linked) nucleotides, 2'-F-arabinonucleotides, 5'-Me, 2'-fluoro nucleotides, morpholino nucleotides, vinylphosphonate deoxyribonucleotides, vinylphosphonate-containing nucleotides, and cyclopropylphosphonate-containing nucleotides (cPrpN). The 2'-modified nucleotides of the present invention (i.e., nucleotides having a group other than a hydroxyl group at the 2' position of the five-membered sugar ring) include, but are not limited to, 2'-O-methyl nucleotides, 2'-deoxy-2'-fluoro nucleotides, 2'-deoxy nucleotides, 2'-methoxyethyl (2'-O-2-methoxyethyl) nucleotides, 2'-amino nucleotides, and 2'-alkyl nucleotides. More than one modification can be incorporated into a single LPA siRNA or even into a single nucleotide thereof. LPA siRNA sense and antisense strands can be synthesized and / or modified by methods known in the art. Modifications at one nucleotide of the present invention are independent of modifications at another nucleotide. Modified nucleotides also include nucleotides having modified nucleobases.

[0150] Locked nucleic acid (LNA) is a type of modified nucleoside that has a methylene bridge added between the C4' and O2' ends of the ribose sugar.

[0151] Glycol nucleic acid (GNA), which has a chiral non-cyclic three-carbon backbone linked by phosphates, is the simplest phosphodiester-based nucleic acid analog.

[0152] The term "sequence" or "nucleotide sequence" refers to a series or order of nucleobases, nucleotides and / or nucleosides, whether modified or unmodified, described as a series of letters using standard nucleotide nomenclature and the symbols for modified nucleotides described herein.

[0153] The term "blunt end" refers to the ends of a double-stranded siRNA where the terminal nucleotides of the two annealed strands are complementary (form complementary base pairs).

[0154] The term "diskle end" refers to the end of a double-stranded siRNA in which the terminal nucleotides of the two annealed strands form a pair (i.e., do not form an overhang) but are not complementary (i.e., form a non-complementary pair). In some embodiments, the 5' end of the sense strand and the 3' end of the antisense strand of the siRNA form a diskle end. In some embodiments, the 3' end of the sense strand and the 5' end of the antisense strand of the siRNA form a diskle end. In some embodiments, both ends of the siRNA form a diskle end. In some embodiments, neither end of the siRNA is a diskle end.

[0155] The term "overhang" refers to a stretch of one or more unpaired nucleotides at the end of one strand of a double-stranded siRNA. The unpaired nucleotides can be on the sense or antisense strand, resulting in a 3' or 5' overhang. In some embodiments, the siRNA contains: a blunt end and a frayed end, a blunt end and a 5' overhang end, a blunt end and a 3' overhang end, a frayed end and a 5' overhang end, a frayed end and a 3' overhang end, two 5' overhang ends, two 3' overhang ends, a 5' overhang end and a 3' overhang end, two frayed ends, or two blunt ends.

[0156] The term "extension" refers to 1, 2, 3, 4, 5, or 6 nucleotides included at the 5' and / or 3' ends of the sense strand core sequence and / or antisense strand core sequence. The extension nucleotides on the sense strand may or may not be complementary to the nucleotides in the corresponding antisense strand (core sequence nucleotides or extension nucleotides). Conversely, the extension nucleotides on the antisense strand may or may not be complementary to the nucleotides in the corresponding sense strand (core sequence nucleotides or extension nucleotides). In some embodiments, both the sense and antisense strands of the siRNA contain 3' and 5' extensions. In some embodiments, the 3' extension nucleotides of one strand base pair with the 5' extension nucleotides of the other strand. In other embodiments, the 3' extension nucleotides of one strand do not base pair with the 5' extension nucleotides of the other strand. In some embodiments, the LPA siRNA has an antisense strand with a 3' extension and a sense strand with a 5' extension.

[0157] The term "complementary" when used to describe a first nucleotide sequence (such as an siRNA sense strand or LPA mRNA) with respect to a second nucleotide sequence (such as an siRNA antisense strand) refers to the ability of nucleotides comprising the first nucleotide sequence to hybridize (form base pair hydrogen bonds) with nucleotides comprising the second nucleotide sequence under certain conditions and form a duplex or double helical structure. Complementary sequences include Watson-Crick base pairs or non-Watson-Crick base pairs, and include natural or modified nucleotides or nucleotide mimetics, as long as the above requirements regarding their hybridization ability are met.

[0158] The term "fully complementary" means that all (100%) of the bases in a contiguous sequence of a first polynucleotide hybridize to the same number of bases in a contiguous sequence of a second polynucleotide.

[0159] The term "partially complementary" means that in a hybridizing nucleobase sequence pair, at least 70% (e.g., 70%, 75%, 80%, 85%, 90%, 95% or 100%) of the bases in the contiguous sequence of the first polynucleotide will hybridize to the same number of bases in the contiguous sequence of the second polynucleotide.

[0160] The term "substantially complementary" means that in a hybridizing nucleobase sequence pair, at least 85% (e.g., 85%, 90%, 95% or 100%) of the bases in the contiguous sequence of the first polynucleotide will hybridize to the same number of bases in the contiguous sequence of the second polynucleotide.

[0161] The terms "complementary," "fully complementary," and "substantially complementary" can be used with respect to base matching between the sense and antisense strands of an siRNA, or between the antisense strand of an siRNA and the sequence of LPA mRNA. Sequence identity or complementarity is independent of modification. For the purpose of determining identity or complementarity, for example, a and Af are complementary to U (or T) and identical to A.

[0162] The term "LPA" may refer to the LPA gene, LPA mRNA or LP(a) protein, as appropriate.

[0163] The terms "silencing," "reducing," "inhibiting," "downregulating," or "knockdown gene expression," when referring to an LPA gene, mean that when a cell, cell population, or tissue is treated with the LPA siRNA, the expression of the gene (as measured by the level of RNA transcribed from the gene or the level of polypeptide, protein, or protein subunit translated from the mRNA in the cell, cell population, or tissue in which the LPA gene is transcribed) is reduced. In some embodiments, the gene expression level and / or mRNA level of LPA in a subject administered the LPA siRNA is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98% relative to the subject before administration of the LPA siRNA or a subject that did not receive the LPA siRNA. Gene expression levels and / or mRNA levels in a subject can be reduced in cells, cell populations, and / or tissues of the subject. In some embodiments, the protein level of LPA in a subject administered the LPA siRNA is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98% relative to the subject before administration of the LPA siRNA or a subject that did not receive the LPA siRNA. Protein levels in a subject can be reduced in cells, cell populations, tissues, blood, and / or other fluids of the subject. Reduction in gene expression, mRNA, or protein levels can be assessed by any method known in the art. A reduction or decrease in LPA mRNA levels and / or protein levels is collectively referred to herein as a decrease or reduction in LPA, or as an inhibition or reduction in the expression of LPA.

[0164] The term "targeting group" includes, but is not limited to, compounds with affinity for cell surface molecules, cell receptor ligands, haptens, antibodies, monoclonal antibodies, antibody fragments, and antibody mimetics with affinity for somatic cell surface molecules. The targeting group can be monovalent, divalent, trivalent, tetravalent, or have a higher valence. In some embodiments, the targeting group is connected to the siRNA using a linker such as a PEG linker or one, two, or three abasic and / or ribitol groups. In some embodiments, the targeting group includes a galactose cluster. The LPA siRNA described herein having a reactive group (e.g., an amine group) at the 5'-end can be synthesized. The reactive group can be used to subsequently attach a targeting moiety using typical methods in the art. In some embodiments, the targeting group includes an asialoglycoprotein receptor ligand. In some embodiments, the asialoglycoprotein receptor ligand includes or consists of one or more galactose derivatives or galactose clusters.

[0165] The term "galactose cluster" includes molecules having two to four terminal galactose derivatives. The terminal galactose derivative is attached to the molecule via its C-1 carbon. In some embodiments, the galactose cluster is a galactose derivative trimer, a three-touch galactose derivative, or a trivalent galactose derivative. In some embodiments, the galactose cluster includes N-acetylgalactosamine (GalNAc). In some embodiments, the galactose cluster includes trivalent N-acetylgalactosamine. In some embodiments, the galactose cluster is a galactose derivative tetramer, a four-touch galactose derivative, or a tetravalent galactose derivative. In some embodiments, the galactose cluster includes tetravalent N-acetylgalactosamine.

[0166] The term "galactose derivative" includes galactose, and galactose derivatives having an affinity for the asialoglycoprotein receptor (equal to or greater than that of galactose). Galactose derivatives include, but are not limited to, galactose, galactosamine, N-formylgalactosamine, N-acetylgalactosamine, N-propionylgalactosamine, N-n-butyrylgalactosamine, and N-isobutyrylgalactosamine. Galactose derivatives and galactose clusters that can be used to target oligonucleotides and other molecules to the liver in vivo are known in the art. Galactose derivatives have been used to target molecules to hepatocytes in vivo by binding to the asialoglycoprotein receptor (ASGPR) expressed on the surface of hepatocytes. Binding of ASGPR ligands to ASGPR promotes cell-specific targeting to hepatocytes and endocytosis of molecules into hepatocytes. Galactose clusters can be attached to the 3' or 5' end of the polynucleotide of the siRNA using methods known in the art.

[0167] The term "galactose derivative trimer" contains three galactose derivatives each attached to a central branch point. In some embodiments, the galactose cluster consists of a galactose derivative trimer that is an N-acetylgalactosamine trimer.

[0168] The term "galactose derivative tetramer" refers to a galactose cluster comprising four galactose derivatives each attached to a central branch point. In some embodiments, the galactose cluster consists of a galactose derivative tetramer that is an N-acetylgalactosamine tetramer.

[0169] The term "linker or spacer" refers to a group that connects the galactose derivative to a branch point or attaches a branch point to the siRNA.

[0170] The term "branch point" refers to any small molecule that allows for the attachment of multiple (eg, three or four) galactose derivatives and further allows for the attachment of the branch point to the siRNA.

[0171] The term "linker" refers to a linker or connecting group that facilitates the covalent linkage of the LPA siRNA to a targeting group, a delivery polymer, or a delivery vehicle. The linker can be attached to the 3' or 5' end of the siRNA sense or antisense strand. In some embodiments, the linker is attached to the siRNA sense strand. In some embodiments, the linker is conjugated to the 5' or 3' end of the siRNA sense strand. In some embodiments, the linker is conjugated to the 5' end of the siRNA sense strand.

[0172] A linker or a linking group is a connection between two atoms that connects a target chemical group (such as siRNA) or a segment to another target chemical group (such as a targeting group or a delivery polymer) or a segment via one or two covalent bonds. Unstable linkages contain unstable bonds. The linkage may optionally include a spacer that increases the distance between the two linked atoms. The spacer may also increase flexibility and / or length for the linkage. The spacer may include, but is not limited to, an alkyl group, an alkenyl group, an alkynyl group, an aryl group, an aralkyl group, an aralkenyl group, and an aralkynyl group; each of which may contain one or more heteroatoms, heterocycles, amino acids, nucleotides, and sugars. Spacer groups are well known in the art, and the foregoing list is not intended to limit the scope of this specification.

[0173] The term "delivery vehicle" refers to a substance that can be used to deliver siRNA to cells or tissues. A delivery vehicle is a compound that improves the delivery of siRNA to cells or tissues. In some embodiments, siRNA can be combined with lipids, nanoparticles, polymers, liposomes, micelles, or other delivery systems available in the art. siRNA can also be chemically conjugated to targeting groups, lipids (including but not limited to cholesterol and cholesterol derivatives), nanoparticles, polymers, liposomes, micelles, or other delivery systems available in the art. In some embodiments, LPA siRNA is linked to a targeting ligand comprising an asialoglycoprotein ligand. In some embodiments, LPA siRNA is linked to a targeting ligand comprising or consisting of a galactose cluster.

[0174] The term "pharmaceutical composition" includes a pharmacologically effective amount of at least one LPA siRNA described herein and one or more pharmaceutically acceptable excipients. A pharmaceutically acceptable excipient is a substance other than the active pharmaceutical ingredient (API, therapeutic product, such as LPA siRNA) that has undergone appropriate safety evaluation and is intentionally included in a drug delivery system. Excipients do not exert or are not intended to exert a therapeutic effect at the intended dose. The siRNAs and pharmaceutical compositions comprising the LPA siRNAs disclosed herein can be packaged or included in a kit, container, package, or dispenser. The LPA siRNAs and pharmaceutical compositions comprising the LPA siRNAs can be packaged in prefilled syringes or vials. These pharmaceutical compositions can be used to inhibit LPA gene expression in cells, tissues, or organisms. In some embodiments, the pharmaceutical compositions are used to treat a subject suffering from a disease, disorder, or condition that would benefit from reduced or inhibited LPA expression. In some embodiments, the pharmaceutical compositions are used to treat a subject at risk of developing a disease, disorder, or condition that would benefit from reduced or inhibited LPA expression. Diseases, disorders or conditions that would benefit from reduced or inhibited LPA expression include, but are not limited to, Buerger's disease, peripheral arterial disease, coronary artery disease, metabolic syndrome, acute coronary syndrome, aortic stenosis, aortic regurgitation, aortic dissection, retinal artery occlusion, cerebrovascular disease, mesenteric ischemia, superior mesenteric artery occlusion, renal artery stenosis, stable / unstable angina, acute coronary syndrome, heterozygous or homozygous familial hypercholesterolemia, hyperlipoprotein betalipoproteinemia, cerebrovascular atherosclerosis, cerebrovascular disease and venous thrombosis. In some embodiments, the subject is a mammal, including but not limited to a human patient.

[0175] The siRNA is delivered to cells, tissues, human organisms, or non-human organisms by any means available in the art. In some embodiments, the cells are mammalian cells, including but not limited to human cells. The cells, tissues, or non-human organisms can be used for research or as research tools (e.g., drug testing or diagnosis).

[0176] The LPA siRNA described herein can be used to treat a subject having, or at risk for, a disease, disorder, or condition that would benefit from reduced or inhibited LPA expression. Treatment of a subject that would benefit from reduced and / or inhibited LPA gene expression includes therapeutic and / or prophylactic treatment. Examples of diseases, disorders, or conditions include, but are not limited to, Buerger's disease, peripheral artery disease, coronary artery disease, metabolic syndrome, acute coronary syndrome, aortic stenosis, aortic regurgitation, aortic dissection, retinal artery occlusion, cerebrovascular disease, mesenteric ischemia, superior mesenteric artery occlusion, renal artery stenosis, stable / unstable angina, acute coronary syndrome, heterozygous or homozygous familial hypercholesterolemia, hyperlipoprotein betalipoproteinemia, cerebral atherosclerosis, cerebrovascular disease, and venous thrombosis. In some embodiments, the method comprises administering a composition, such as a pharmaceutical composition comprising the LPA siRNA described herein, to the mammal to be treated.

[0177] In some embodiments, a therapeutically effective amount of one or more of the LPA siRNAs is administered to a subject to inhibit the expression of LPA in the subject (e.g., an amount effective to inhibit the expression of LPA in the subject). In some embodiments, one or more of the LPA siRNAs described herein are used to treat a subject suffering from a disease or condition that would benefit from a reduction or inhibition of LPA expression. In some embodiments, the LPA siRNAs are used to treat or prevent at least one symptom in a subject suffering from a disease or condition that would benefit from a reduction or inhibition of LPA expression. A therapeutically effective amount of any one or more of the siRNAs is administered to a subject to treat a symptom. In some embodiments, a prophylactically effective amount of any one or more of the siRNAs is administered to a subject to prevent at least one symptom.

[0178] In some embodiments, LPA siRNA is used to treat or manage clinical manifestations, wherein a therapeutically or prophylactically effective amount of one or more LPA siRNAs or compositions comprising LPA siRNAs described herein is administered to a subject in need of such treatment, prevention, or management. In some embodiments, the method comprises administering a composition comprising LPA siRNAs described herein to a mammal to be treated.

[0179] The route of administration is the route by which the LPA siRNA comes into contact with the body. Generally, methods for administering drugs and nucleic acids used to treat subjects are well known in the art and can be applied to the administration of the compositions described herein. The compounds described herein can be administered via any suitable route in formulations appropriately tailored for that particular route. Thus, the compounds described herein can be administered by injection, for example, intravenously, intramuscularly, intradermally, subcutaneously, or intraperitoneally. BRIEF DESCRIPTION OF THE DRAWINGS

[0180] The features and advantages of the present invention can be better understood by referring to the exemplary embodiments described in detail below and the accompanying drawings. The accompanying drawings are briefly described as follows:

[0181] FIG1 is a flow chart of the solid phase phosphoramidite technique for synthesizing the LPA siRNA agent of the present invention.

[0182] FIG2 is a graph showing the effect of the RNA inhibitors in Table 7 prepared in Example 4 of the present invention on reducing Lp(a) levels in the serum of transgenic mice at different concentrations;

[0183] FIG3 is a graph showing the effect of the RNA inhibitors in Table 8 prepared in Example 5 of the present invention on reducing Lp(a) levels in the serum of transgenic mice at different concentrations;

[0184] FIG4 is a graph showing the effect of the RNA inhibitors in Table 9 prepared in Example 6 of the present invention on reducing Lp(a) levels in cynomolgus monkey serum. DETAILED DESCRIPTION

[0185] The present invention is further described below with reference to examples.

[0186] Example 1 Design and Synthesis of LPA siRNA

[0187] The present invention designed small interfering nucleic acids (siRNAs) as shown in Table 1 for the lipoprotein gene (transcript information: NM005577.4). The transcript information of this protein gene is recorded in the NCBI database. Table 1 lists the sequences of the siRNAs targeting LPA mRNA and their sense and antisense strands. Table 2 lists the siRNAs and their sense and antisense strands after adding two dTs to the 3' ends of the sense and antisense strands based on Table 1. The present invention then modified the above sequences, and the modified siRNAs, their sense and antisense strands are shown in Table 3. siRNAs modified with targeting groups, their sense and antisense strands are shown in Table 4. siRNAs and their sense and antisense strands disclosed in the prior art are also shown in Table 4.

[0188] The LPA siRNA agent of the present invention can be synthesized using conventional solid-phase phosphoramidite technology, as shown in Figure 1. All materials are commercially available. Briefly, the DMT protection is first removed using TCA, followed by washing with ACN. Coupling, capping, oxidation / thiolation, and ACN washing are then performed, followed by repeated coupling cycles. After synthesizing the target sequence, aminolysis is performed using aqueous ammonia at 40°C for 16 hours, followed by centrifugation and drying. The product is then desalted or purified by HPLC. For UPLC / TOF-MS analysis and UV quantification, a 4-μL sample is added to 196 μL of water onto a UV plate. Concentration is measured using a microplate reader and the nmol concentration is calculated. Annealing: The antisense strand:sense strand ratio is 1.05:1. The mixture is then aliquoted into 2.5 nmol x 2 tubes (suitable for centrifuge tubes). The tubes are then lyophilized individually, ensuring minimal solution volume. The tubes are sealed with aluminum foil during lyophilization to yield the final siRNA.

[0189] Example 2 Detection of LPA expression level

[0190] LPA expression levels were assessed using a dual-luciferase reporter assay. The psiCHECK2-LPA dual-luciferase reporter plasmid was first constructed, inserting the full-length LPA sequence into the psiCHECK2-LPA vector. The LPA sequence was obtained from Origene, and the psiCHECK2 vector was obtained from Promega. HUH7 cells were seeded in 96-well plates and cultured in a 37°C incubator for 2 hours to allow attachment. The LPA dual-reporter plasmid was then transfected into the HUH7 cells using FugeneHD and incubated overnight at 37°C in a 5% CO2 incubator. After 16 hours of overnight incubation, HUH7 cells were transfected with LPA-siRNA using RNAiMAX (Invitrogen) and cultured for 24 hours. After 24 hours of incubation in a 5% CO2 incubator at 37°C, the expression of Firefly luciferase and Renilla luciferase was assessed using the Dual-Glo luciferase Assay System. The calculation formulas for the absolute and relative inhibition rates of LPA are as follows, where the sample represents the group with 0.3 nM siRNA added to the test, the internal reference represents the group without siRNA added, and the control represents the group with 0.3 nM control siRNA added.

[0191] LPA inhibition rate (%) = {1-[(Renilla lum of sample - average value of background Renilla lum) / (Firefly lum of sample - average value of background Firefly lum)] / [(Renilla lum of internal reference - average value of background Renilla lum) / (Firefly lum of internal reference - average value of background Firefly lum)]} × 100%. The LPA inhibition rate of the control siRNA was 60.38%.

[0192] LPA relative inhibition rate (%) = (sample LPA inhibition rate / control LPA inhibition rate) × 100%.

[0193] The results are shown in Table 5 below:

[0194] Table 5

[0195] Example 3 In vitro experiments of LPA RNAi agents

[0196] LPA expression levels were evaluated using a dual-luciferase reporter assay. First, the psiCHECK2-LPA dual-luciferase reporter plasmid was constructed by inserting the full-length human LPA sequence into the psiCHECK2 vector (from Origene). The LPA sequence was obtained from Origene, and the psiCHECK2 vector was obtained from Promega. Hep3B cells were seeded at 10,000 cells per well in a 96-well plate and cultured overnight at 37°C to allow attachment. The LPA reporter plasmid was transfected into the cells using 0.2 μL / well of FUGENE HD (Promega, E2311). After incubation for 6 hours, the cells were transfected with siRNA using 0.2 μL / well of lipoFectamine 2000 (Thermo, 11668-019). The cells were incubated at 37°C with 5% CO2 for 24 hours. The expression of Firefly luciferase and Renilla luciferase was detected using the Dual-Glo Luciferase Assay System (Promega E29440) to determine the expression level of LPA.

[0197] LPA inhibition rate (%) = {1-[(average value of Renilla lum of sample - average value of background Renilla lum) / (average value of Firefly lum of sample - average value of background Firefly lum)] / [(average value of Renilla lum of internal control - average value of background Renilla lum) / (average value of Firefly lum of internal control - average value of background Firefly lum)]}×100%. IC was calculated using the dose-response (variable slope) four-parameter method in GraphPad Prism software. 50 The results are shown in Table 6 below:

[0198] Table 6

[0199] Example 4 In vivo testing of the efficacy of LPA RNAi agents in transgenic mice

[0200] To evaluate the in vivo activity of the test siRNA, this experiment used LPA humanized transgenic mice to evaluate the in vivo activity of the test molecules. Transgenic mice were subcutaneously injected with 3 mg / kg of JAB-190024-1, JAB-190024-3, JAB-190024-5 and 1 mg / kg of JAB-190024-1, JAB-190024-3, JAB-190024-5, SLN360 on day 0, and the control group mice were given an equal volume of phosphate buffered saline (PBS). Mouse serum was collected before administration and on days 7, 14, 28, 42, and 56 after administration. The human lipoprotein (a) Elisa (Abcam ab212165) kit was used to detect the level of Lp (a) in mouse serum. Normalization was performed based on the Lp (a) level in the serum samples from the PBS-treated group to calculate the relative level of Lp (a) in the drug-treated group. The results are shown in Table 7 and Figure 2:

[0201] Table 7

[0202] Example 5 In vivo testing of the efficacy of LPA RNAi agents in transgenic mice

[0203] To evaluate the in vivo activity of the test siRNAs, LPA humanized transgenic mice were used to evaluate the in vivo activity of the test molecules. Transgenic mice were subcutaneously injected with 1 mg / kg of AMG890, SLN360, JAB-190024-1, JAB-190024-2, JAB-190024-6, JAB-190024-4 and 0.3 mg / kg of JAB-190024-1, JAB-190024-2, JAB-190024-6, JAB-190024-4 on day 0. Control mice were given an equal volume of phosphate-buffered saline (PBS). Mouse serum was collected on the day before dosing (Day-1) and on days 7, 14, 28, 42, 56, and 84 after dosing. Human Lp(a) levels in mouse serum were measured using a Human Lipoprotein(a) Elisa (Abcam, ab212165). The relative Lp(a) levels in the dosing groups were calculated based on the Lp(a) levels in serum samples from the PBS-treated group. The results are shown in Table 8 and Figure 3:

[0204] Table 8

[0205] Example 6 In vivo testing of the efficacy of cynomolgus monkey LPA RNAi agents

[0206] To evaluate the in vivo activity of the test siRNAs in large animals, this study used cynomolgus macaques (male, 4-7 years old, weighing 3-6 kg) to evaluate the in vivo activity of the test molecules. The macaques were housed in an animal room controlled at a temperature of 18°C ​​to 26°C and a relative humidity of 40% to 70%. The macaques were subcutaneously injected with 1 mg / kg of JAB-190024-1, JAB-190024-2, or JAB-190024-6. Serum was collected before dosing and on days 7, 14, and 21 after dosing. 0.5 mL of serum was aliquoted into two EP tubes and stored at -80°C for subsequent Lp(a) level analysis. Serum Lp(a) levels were measured using the Mercodia Lp(a) Elisa kit (Mercodia, 10-1106-01). Normalization was performed based on the serum Lp(a) levels in cynomolgus monkeys before administration to calculate the relative Lp(a) levels in each group at different time points after administration. The results are shown in Table 9 and Figure 4.

[0207] Table 9

[0208] Example 7

[0209] The experiments on nucleic acid-protein interactions were based on the surface plasmon resonance principle and were performed using a GE Biacore T200 instrument and a Cytiva S series SA chip.

[0210] According to the supplier's manual, the biotinylated AS chain RNA ligand was coupled to the channel surface of the Sensor Chip SA chip (Cytiva, catalog number BR-1005-31) using the standard ligand nucleic acid coupling procedure, and the target coupling amount was set to 500RU.

[0211] For kinetic measurements, Recombinant Human Argonaute-2 / AGO2 Protein (His Tag) (SinoBiological, catalog number 11079-H07B) was applied in increasing concentrations at 25°C in 1X HBS-EP+ buffer with a contact time of 180 seconds and a flow rate of 30 μL / min, and the signal was monitored over time. Specifically, the sample solution was injected onto the chip surface at a flow rate of 30 μL / min for 180 seconds. After each binding reaction, it was naturally dissociated for 1200 seconds, and 0.5% SDS was injected as a regeneration reagent at a flow rate of 30 μL / min for 30 seconds to allow the signal to return to the baseline. A 1:1 binding model was used to fit the data. The sequences tested and the results obtained are shown in Tables 10 and 11 below, respectively.

[0212] Table 10

[0213] Table 11

[0214] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. An siRNA or a pharmaceutically acceptable salt thereof, comprising a sense strand and an antisense strand, wherein the antisense strand comprises a sequence complementary to an LPA mRNA sequence, and the sequence of the antisense strand comprises the sequence of any antisense strand in Table 1.

2. The siRNA or a pharmaceutically acceptable salt thereof according to claim 1, wherein The antisense strand is 19-26 nucleobases long.

3. The siRNA or a pharmaceutically acceptable salt thereof according to claim 2, wherein: The antisense strand is 19-21 nucleobases long.

4. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein: The sense strand includes a sequence that is substantially complementary to the antisense strand and forms a duplex complementary region of 15 to 26 consecutive base pairs.

5. The siRNA or a pharmaceutically acceptable salt thereof according to claim 4, wherein: The length of the duplex complementary region is 18 to 23 consecutive base pairs.

6. The siRNA or a pharmaceutically acceptable salt thereof according to claim 5, wherein: The length of the duplex complementary region is 19 to 21 consecutive base pairs.

7. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein: The length of the sense strand is 19-26 nucleobases.

8. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein: The length of the sense strand is 19-21 nucleobases.

9. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 8, wherein: The sequence of the sense chain includes any sense chain sequence in Table 1.

10. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, wherein: The antisense strand includes any one of SEQ ID NOs. 128-150 and 253-254.

11. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 10, wherein: The positive strand includes any one of SEQ ID NOs. 1-23, 126-127.

12. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, wherein: The siRNA includes any siRNA in Table 1.

13. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 12, wherein: In the siRNA: The antisense strand comprises SEQ ID NO.128, and the sense strand comprises SEQ ID NO.1; The antisense strand includes SEQ ID NO.129, and the sense strand includes SEQ ID NO.2; The antisense strand comprises SEQ ID NO.130, and the sense strand comprises SEQ ID NO.3; The antisense strand includes SEQ ID NO.131, and the sense strand includes SEQ ID NO.4; The antisense strand includes SEQ ID NO.132, and the sense strand includes SEQ ID NO.5; The antisense strand includes SEQ ID NO.133, and the sense strand includes SEQ ID NO.6; The antisense strand includes SEQ ID NO.134, and the sense strand includes SEQ ID NO.7; The antisense strand comprises SEQ ID NO.135, and the sense strand comprises SEQ ID NO.8; The antisense strand comprises SEQ ID NO.136, and the sense strand comprises SEQ ID NO.9; The antisense strand comprises SEQ ID NO.137, and the sense strand comprises SEQ ID NO.10; The antisense strand includes SEQ ID NO.138, and the sense strand includes SEQ ID NO.11; The antisense strand includes SEQ ID NO.139, and the sense strand includes SEQ ID NO.12; The antisense strand comprises SEQ ID NO.140, and the sense strand comprises SEQ ID NO.13; The antisense strand includes SEQ ID NO.141, and the sense strand includes SEQ ID NO.14; The antisense strand includes SEQ ID NO.142, and the sense strand includes SEQ ID NO.15; The antisense strand includes SEQ ID NO.143, and the sense strand includes SEQ ID NO.16; The antisense strand includes SEQ ID NO.144, and the sense strand includes SEQ ID NO.17; The antisense strand comprises SEQ ID NO.145, and the sense strand comprises SEQ ID NO.18; The antisense strand comprises SEQ ID NO.146, and the sense strand comprises SEQ ID NO.19; The antisense strand comprises SEQ ID NO.147, and the sense strand comprises SEQ ID NO.20; The antisense strand comprises SEQ ID NO.148, and the sense strand comprises SEQ ID NO.21; The antisense strand includes SEQ ID NO.149, and the sense strand includes SEQ ID NO.22; The antisense strand comprises SEQ ID NO.150, and the sense strand comprises SEQ ID NO.23; The antisense strand includes SEQ ID NO.253, and the sense strand includes SEQ ID NO.126 or The antisense strand includes SEQ ID NO.254, and the sense strand includes SEQ ID NO.

127.

14. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 13, wherein: The antisense strand or sense strand contains 1, 2, 3, 4, 5 or 6 stretches.

15. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 14, wherein: The extension of the antisense strand or sense strand is dT.

16. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 15, wherein: The antisense strand or sense strand contains 2 dT stretches.

17. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 15, wherein: The 3' end of the antisense strand contains two extended dTs, and the sense strand contains no dT.

18. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 17, wherein: The antisense strand includes the sequence of any antisense strand in Table 2.

19. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 18, wherein: The antisense strand includes any one of SEQ ID NOs. 380-402 and 544-546.

20. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 17, wherein: The sense strand includes the sequence of any sense strand in Table 2.

21. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 20, wherein: The positive strand includes any one of SEQ ID NOs. 255-277 and 541-543.

22. The siRNA or pharmaceutically acceptable salt thereof according to any one of claims 1 to 21, wherein the siRNA comprises any one of the siRNAs in Table 2.

23. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 22, wherein: The antisense strand includes SEQ ID NO.380, and the sense strand includes SEQ ID NO.255; The antisense strand includes SEQ ID NO.381, and the sense strand includes SEQ ID NO.256; The antisense strand includes SEQ ID NO.382, and the sense strand includes SEQ ID NO.257; The antisense strand includes SEQ ID NO.383, and the sense strand includes SEQ ID NO.258; The antisense strand includes SEQ ID NO.384, and the sense strand includes SEQ ID NO.259; The antisense strand includes SEQ ID NO.385, and the sense strand includes SEQ ID NO.260; The antisense strand includes SEQ ID NO.386, and the sense strand includes SEQ ID NO.261; The antisense strand includes SEQ ID NO.387, and the sense strand includes SEQ ID NO.262; The antisense strand includes SEQ ID NO.388, and the sense strand includes SEQ ID NO.263; The antisense strand includes SEQ ID NO.389, and the sense strand includes SEQ ID NO.264; The antisense strand includes SEQ ID NO.390, and the sense strand includes SEQ ID NO.265; The antisense strand includes SEQ ID NO.391, and the sense strand includes SEQ ID NO.266; The antisense strand includes SEQ ID NO.392, and the sense strand includes SEQ ID NO.267; The antisense strand includes SEQ ID NO.393, and the sense strand includes SEQ ID NO.268; The antisense strand includes SEQ ID NO.394, and the sense strand includes SEQ ID NO.269; The antisense strand includes SEQ ID NO.395, and the sense strand includes SEQ ID NO.270; The antisense strand includes SEQ ID NO.396, and the sense strand includes SEQ ID NO.271; The antisense strand includes SEQ ID NO.397, and the sense strand includes SEQ ID NO.272; The antisense strand includes SEQ ID NO.398, and the sense strand includes SEQ ID NO.273; The antisense strand includes SEQ ID NO.399, and the sense strand includes SEQ ID NO.274; The antisense strand comprises SEQ ID NO.400, and the sense strand comprises SEQ ID NO.275; The antisense strand includes SEQ ID NO.401, and the sense strand includes SEQ ID NO.276; The antisense strand comprises SEQ ID NO.402, and the sense strand comprises SEQ ID NO.277; The antisense strand includes SEQ ID NO.541, and the sense strand includes SEQ ID NO.544; The antisense strand comprises SEQ ID NO.542, and the sense strand comprises SEQ ID NO.545; or The antisense strand includes SEQ ID NO.543, and the sense strand includes SEQ ID NO.

546.

24. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 23, wherein: The sense strand and / or antisense strand independently comprises one or more modified nucleotides 25. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 24, wherein: The modified nucleotides are 2'-modified nucleotides.

26. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 25, wherein: The modified nucleotides include: 2'-O-methyl modified nucleotides, 2'-O-methoxyethyl modified nucleotides, 2'-fluorine modified nucleotides, 2'-deoxy modified nucleotides, 2'-O-alkenyl modified nucleotides, locked nucleotides, GNA, LNA, abasic nucleotides, deoxythymidine, inverted deoxythymidine, deoxyadenosine, inverted deoxyadenosine, 2'-amino-modified nucleotides, 2'-alkyl-modified nucleotides, morpholino nucleotides and nucleotides including non-natural bases, 5'-(E)-vinyl phosphate modified nucleotides, or combinations thereof.

27. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 26, wherein: The nucleotides described are all modified nucleotides.

28. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 27, wherein: The modified nucleotides include 2'-O-methyl modified nucleotides, 2'-fluorine modified nucleotides, deoxythymidine and 5'-(E)-vinyl phosphate modified nucleotides, or a combination thereof.

29. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 28, wherein: The sense strand, antisense strand, or both the sense strand and antisense strand include one or more phosphorothioate diester or phosphorothioate diester internucleoside linkages.

30. The siRNA or a pharmaceutically acceptable salt thereof according to claim 29, wherein: The antisense strand comprises two consecutive monothioate phosphorodiester internucleoside linkages between the 3' and 5' terminal nucleotides.

31. The siRNA or a pharmaceutically acceptable salt thereof according to claim 29 or 30, wherein: The sense strand comprises two consecutive monothioate phosphorodiester internucleoside linkages between the 3' and 5' terminal nucleotides.

32. The siRNA or pharmaceutically acceptable salt thereof according to claim 29 or 30, wherein: The 5'-end terminal nucleotides of the sense strand include two consecutive monothiophosphate diester nucleoside bonds, and the 3'-end terminal nucleotides of the sense strand include one monothiophosphate diester nucleoside bond.

33. The siRNA according to claim 29 or 30 or a pharmaceutically acceptable salt thereof, wherein the 5' terminal nucleotides of the sense strand include two consecutive monothiophosphate diester nucleoside bonds, the 3' terminal nucleotides of the sense strand include one monothiophosphate diester nucleoside bond; and the antisense strand includes two consecutive monothiophosphate diester nucleoside bonds between the 3' and 5' terminal nucleotides.

34. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 33, wherein: 1) The sense strand comprises 21 nucleotides, the antisense strand comprises 23 nucleotides, the length of the duplex complementary region is 21 base pairs, the 9th, 10th and 11th nucleotides from the 5' end of the sense strand are all 2'-fluorine-modified nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl-modified nucleotides; the 8th, 10th, 18th and 22nd nucleotides from the 3' end of the antisense strand are all 2'-fluorine-modified nucleotides, and the remaining nucleotides of the antisense strand are all 2'-O-methyl-modified nucleotides; the 1st to 3rd nucleotides from the 5' end of the sense strand, the 1st to 3rd nucleotides from the 3' end of the sense strand, the 1st to 3rd nucleotides from the 5' end of the antisense strand, and the 1st to 3rd nucleotides from the 3' end of the antisense strand are all connected by monothiophosphate diester bonds, and the remaining nucleotides of the sense strand and the antisense strand are connected by phosphodiester bonds; 2) The sense strand comprises 19 nucleotides, the antisense strand comprises 21 nucleotides, the length of the duplex complementary region is 19 base pairs, the 7th, 8th and 9th nucleotides of the sense strand starting from the 5' end are all 2'-fluorine-modified nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl-modified nucleotides; the 1st and 2nd nucleotides of the antisense strand starting from the 3' end are two dTs, and the 6th, 8th, 14th, 16th and 20th nucleotides of the antisense strand starting from the 3' end are all 2' -fluorine-modified nucleotides, and the remaining nucleotides of the antisense strand are all 2'-O-methyl-modified nucleotides; the first to third nucleotides from the 5' end of the sense strand, the first to third nucleotides from the 3' end of the sense strand, the first to third nucleotides from the 5' end of the antisense strand, and the first to third nucleotides from the 3' end of the antisense strand are all connected by monothioate phosphorodiester bonds, and the remaining nucleotides of the sense strand and the antisense strand are connected by phosphodiester bonds; 3) The sense strand comprises 19 nucleotides, the antisense strand comprises 21 nucleotides, the length of the duplex complementary region is 19 base pairs, the 7th, 8th and 9th nucleotides of the sense strand starting from the 5' end are all 2'-fluorine-modified nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl-modified nucleotides; the 1st and 2nd nucleotides of the antisense strand starting from the 3' end are two dTs, and the 4th, 6th, 8th, 16th and 20th nucleotides of the antisense strand starting from the 3' end are all 2'- Fluorine-modified nucleotides, the remaining nucleotides of the antisense chain are all 2'-O-methyl-modified nucleotides; the first to third nucleotides from the 5' end of the sense chain, the first to third nucleotides from the 3' end of the sense chain, the first to third nucleotides from the 5' end of the antisense chain, and the first to third nucleotides from the 3' end of the antisense chain are all connected by monothioate phosphorodiester bonds, and the remaining nucleotides of the sense chain and the antisense chain are connected by phosphodiester bonds; 4) The sense strand comprises 21 nucleotides, the antisense strand comprises 21 nucleotides, the length of the duplex complementary region is 21 base pairs, the 9th, 10th and 11th nucleotides of the sense strand from the 5' end are all 2'-fluorine-modified nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl-modified nucleotides; the 4th, 6th, 8th, 10th, 16th, 18th and 20th nucleotides of the antisense strand from the 3' end are all 2'-fluorine-modified nucleotides, and the remaining nucleotides of the antisense strand are modified with 2'-O-methyl-modified nucleotides; the 1st to 3rd nucleotides of the sense strand from the 5' end, the 1st to 3rd nucleotides of the sense strand from the 3' end, the 1st to 3rd nucleotides of the antisense strand from the 5' end, and the 1st to 3rd nucleotides of the antisense strand from the 3' end are all connected by monothiophosphate diester bonds, and the remaining nucleotides of the sense strand and the antisense strand are connected by phosphodiester bonds; 5) The sense strand comprises 21 nucleotides, the antisense strand comprises 21 nucleotides, the length of the duplex complementary region is 21 base pairs, the 9th, 10th and 11th nucleotides of the sense strand starting from the 5' end are all 2'-fluorinated nucleotides, and the sense strand The remaining nucleotides of the antisense chain are all 2'-O-methyl modified nucleotides; the 4th, 6th, 8th, 10th, 16th, 18th and 20th nucleotides from the 3rd position of the 3' end of the antisense chain are all 2'-fluorine modified nucleotides, the remaining nucleotides of the antisense chain are modified with 2'-O-methyl modified nucleotides, and the first nucleotide of the 5' end of the antisense chain is modified with VP; the first to third nucleotides from the 5' end of the sense chain, the first to third nucleotides from the 3' end of the sense chain, the first to third nucleotides from the 5' end of the antisense chain, and the first to third nucleotides from the 3' end of the antisense chain are all connected by monothioate phosphorodiester bonds, and the remaining nucleotides of the sense chain and the antisense chain are connected by phosphodiester bonds; 6) The sense strand comprises 19 nucleotides, the antisense strand comprises 21 nucleotides, the length of the duplex complementary region is 19 base pairs, the 7th, 8th and 9th nucleotides of the sense strand from the 5' end are all 2'-fluorine-modified nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl-modified nucleotides; the 1st and 2nd positions of the antisense strand from the 3' end are two dTs, the 6th, 8th, 14th, 16th and 20th nucleotides of the antisense strand from the 3' end are all 2'-fluorine-modified nucleotides, and the remaining nucleotides of the antisense strand are all 2'-O-methyl-modified nucleotides. The remaining nucleotides of the sense strand are all 2'-O-methyl modified nucleotides, and the first nucleotide at the 5' end of the antisense strand is modified with VP; the first to third nucleotides from the 5' end of the sense strand, the first to third nucleotides from the 3' end of the sense strand, the first to third nucleotides from the 5' end of the antisense strand, and the first to third nucleotides from the 3' end of the antisense strand are all connected by monothioate phosphorodiester bonds, and the remaining nucleotides of the sense strand and the antisense strand are connected by phosphodiester bonds; 7) The sense strand comprises 19 nucleotides, the antisense strand comprises 21 nucleotides, the length of the duplex complementary region is 19 base pairs, the 7th, 8th and 9th nucleotides of the sense strand starting from the 5' end are all 2'-fluorine-modified nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl-modified nucleotides; the 1st and 2nd nucleotides of the antisense strand starting from the 3' end are two dTs, and the 6th, 8th, 14th, 16th and 20th nucleotides of the antisense strand starting from the 3' end are all 2'- Fluorine-modified nucleotides, the remaining nucleotides of the antisense strand are all 2'-O-methyl-modified nucleotides; the first to third nucleotides from the 5' end of the sense strand, the first to second nucleotides from the 3' end of the sense strand, the first to third nucleotides from the 5' end of the antisense strand, and the first to third nucleotides from the 3' end of the antisense strand are all connected by monothioate phosphorodiester bonds, and the remaining nucleotides of the sense strand and the antisense strand are connected by phosphodiester bonds; or 8) The sense strand comprises 19 nucleotides, the antisense strand comprises 21 nucleotides, the length of the duplex complementary region is 19 base pairs, the 7th, 8th and 9th nucleotides of the sense strand from the 5' end are all 2'-fluorine-modified nucleotides, and the remaining nucleotides of the sense strand are all 2'-O-methyl-modified nucleotides; the 1st and 2nd positions of the antisense strand from the 3' end are two dTs, the 6th, 8th, 14th, 16th and 20th nucleotides of the antisense strand from the 3' end are all 2'-fluorine-modified nucleotides, and the remaining nucleotides of the antisense strand are all 2'-O-methyl-modified nucleotides. The remaining nucleotides of the sense strand are all 2'-O-methyl modified nucleotides, and the first nucleotide at the 5' end of the antisense strand is modified with VP; the first to third nucleotides from the 5' end of the sense strand, the first to second nucleotides from the 3' end of the sense strand, the first to third nucleotides from the 5' end of the antisense strand, and the first to third nucleotides from the 3' end of the antisense strand are all connected by monothioate phosphorodiester bonds, and the remaining nucleotides of the sense strand and the antisense strand are connected by phosphodiester bonds.

35. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 34, wherein: The antisense strand includes any one of the antisense strands in Table 3.

36. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 35, wherein: The antisense strand includes any antisense strand of SEQ ID NO. 505, 507, 509, 511, 513, 515, 517, 519, 521, 523, 525 and 527.

37. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 36, wherein: The sense strand includes any sense strand in Table 3.

38. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 37, wherein: The sense strand includes any sense strand of SEQ ID NO. 506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526 and 528.

39. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 38, wherein: The siRNA includes any siRNA in Table 3.

40. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 39, wherein: The antisense strand includes SEQ ID NO.505, and the sense strand includes SEQ ID NO.506; The antisense strand includes SEQ ID NO.507, and the sense strand includes SEQ ID NO.508; The antisense strand comprises SEQ ID NO.509, and the sense strand comprises SEQ ID NO.510; The antisense strand includes SEQ ID NO.511, and the sense strand includes SEQ ID NO.512; The antisense strand includes SEQ ID NO.513, and the sense strand includes SEQ ID NO.514; The antisense strand includes SEQ ID NO.515, and the sense strand includes SEQ ID NO.516; The antisense strand includes SEQ ID NO.517, and the sense strand includes SEQ ID NO.518; The antisense strand includes SEQ ID NO.519, and the sense strand includes SEQ ID NO.520; The antisense strand includes SEQ ID NO.521, and the sense strand includes SEQ ID NO.522; The antisense strand includes SEQ ID NO.523, and the sense strand includes SEQ ID NO.524; The antisense strand comprises SEQ ID NO.525, and the sense strand comprises SEQ ID NO.526; or The antisense strand includes SEQ ID NO.527, and the sense strand includes SEQ ID NO.

528.

41. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 40, wherein: The siRNA further comprises a targeting group.

42. The siRNA or a pharmaceutically acceptable salt thereof according to claim 41, wherein The targeting group includes an asialoglycoprotein receptor ligand.

43. The siRNA or pharmaceutically acceptable salt thereof according to claim 42, wherein: The asialoglycoprotein receptor ligand comprises a galactose cluster.

44. The siRNA or pharmaceutically acceptable salt thereof according to claim 43, wherein: The galactose cluster is a monovalent, divalent, trivalent or tetravalent galactose derivative.

45. The siRNA according to claim 44 or a pharmaceutically acceptable salt thereof, wherein: The galactose derivative is N-acetylgalactosamine (GalNAc).

46. ​​The siRNA or a pharmaceutically acceptable salt thereof according to claim 45, wherein The galactose cluster is trivalent N-acetylgalactosamine.

47. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 46, wherein: The galactose cluster is connected to the branch center through a linker, and the branch center is conjugated to the nucleotide of siRNA through the linker.

48. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 47, wherein: The targeting group is conjugated to the 5' end of the sense strand.

49. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 47, wherein: The targeting group is conjugated to the 3' end of the sense strand.

50. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 47, wherein: The following fragments containing targeting groups are conjugated to the 5' or 3' end of the sense strand:

51. The siRNA according to claim 50 or a pharmaceutically acceptable salt thereof, wherein: The fragment comprising the targeting group is conjugated to the 5' end of the sense strand.

52. The siRNA or a pharmaceutically acceptable salt thereof according to claim 51, wherein The fragment comprising the targeting group is conjugated to the 3' end of the sense strand.

53. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 52, wherein: The siRNA includes any one of the siRNAs in Table 4.

54. The siRNA according to claim 53 or a pharmaceutically acceptable salt thereof, wherein: The sense strand is any sense strand in Table 4.

55. The siRNA according to claim 53 or a pharmaceutically acceptable salt thereof, wherein: The antisense strand is any antisense strand in Table 4.

56. A pharmaceutical composition comprising the siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 55, and a pharmaceutically acceptable salt thereof.

57. A method for reducing the expression level of LPA in a subject, the method comprising administering to the subject the siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 55 or the pharmaceutical composition according to claim 56.

58. Use of the siRNA or pharmaceutically acceptable salt thereof according to any one of claims 1 to 55 or the pharmaceutical composition according to claim 56 in the preparation of a drug for preventing or treating a disease associated with LPA expression.

59. The use according to claim 58, wherein: The disease associated with LPA expression is cardiovascular and cerebrovascular disease.

60. The use according to claim 59, wherein: The cardiovascular and cerebrovascular diseases include stroke, atherosclerosis, thrombosis, coronary heart disease or aortic valve stenosis and any other diseases or pathologies associated with increased levels of Lp(a) particles.

61. A method for preventing or treating a disease associated with LPA expression, the method comprising administering to a subject the siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 55 or the pharmaceutical composition according to claim 56.

62. The method of claim 61, wherein: The disease associated with LPA expression is cardiovascular and cerebrovascular disease.

63. The method of claim 62, wherein: The cardiovascular and cerebrovascular diseases include stroke, atherosclerosis, thrombosis, coronary heart disease or aortic valve stenosis and any other diseases or pathologies associated with increased levels of Lp(a) particles.

64. The siRNA or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 55 or the pharmaceutical composition according to claim 56 for preventing or treating a disease associated with LPA expression.

65. The use according to claim 64, wherein: The disease associated with LPA expression is cardiovascular and cerebrovascular disease.

66. The use according to claim 65, wherein: The cardiovascular and cerebrovascular diseases include stroke, atherosclerosis, thrombosis, coronary heart disease or aortic valve stenosis and any other diseases or pathologies associated with increased levels of Lp(a) particles.

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