Oligonucleotide targeting lipoprotein a and use thereof

Abstract

Provided are an oligonucleotide targeting lipoprotein a and a use thereof. The oligonucleotide can significantly inhibit the gene expression level of lipoprotein a and has a lasting drug efficacy.

Description

Oligonucleotides targeting lipoprotein a and uses thereof Technical Field

[0001] The present disclosure relates to an oligonucleotide, in particular to the inhibition of Lp(a) gene expression and the treatment of cardiovascular diseases and cardiovascular-related diseases. Background Art

[0002] Lipoprotein(a) (Lp(a)) is a specialized low-density lipoprotein composed of two components: apolipoprotein-B (apoB) and apolipoprotein-A (apoA). It is a plasminogen-like glycoprotein. ApoB is structurally and physicochemically similar to LDL-C, while apoA is composed of a carbohydrate-rich protein. Lp(a) particle size varies significantly.

[0003] Studies have found that plasma levels of Lp(a) vary widely between individuals, ranging from 20 mg / dL to >2000 mg / dL across different ethnicities, with almost 20% of individuals experiencing extreme levels. Although the exact mechanism by which Lp(a) induces atherosclerosis remains unclear, this lipoprotein has been found in the arterial walls of atherosclerotic plaques. Lp(a) is associated with foam cell formation and lipid deposition in plaques.

[0004] Lp(a) is a marker of cardiovascular disease risk. Normal Lp(a) levels range from 0.1 to 25 mg / dL, with approximately 25% of the US population having Lp(a) levels of 30 mg / dL or higher. Elevated serum Lp(a) levels are an independent risk factor for cardiovascular disease. Meta-analyses have shown that the risk of cardiovascular disease is roughly linearly associated with increasing Lp(a) levels. Baseline Lp(a) ≥ 30 mg / dL is associated with an increased hazard ratio (HR) for cardiovascular events, independent of other cardiovascular risk factors. Willeit et al. reported that in patients receiving statin therapy, the risk of cardiovascular disease is roughly linearly associated with increasing Lp(a) levels. These data suggest that residual cardiovascular disease risk persists in patients receiving maximally tolerated statin therapy, and elevated Lp(a) levels are a target for intervention to further reduce this residual risk.

[0005] The clinical threshold for Lp(a) is difficult to determine. The European Atherosclerosis Society (EAS) proposes that the optimal Lp(a) level for Caucasians without any cardiovascular risk is below the 80th percentile, approximately <50 mg / dL. The Canadian Cardiovascular Society (CCS) recommends focusing on individuals with Lp(a) levels >30 mg / dL, as this group has an approximately two-fold increased risk of cardiovascular disease.

[0006] Lp(a) screening should be considered for patients with premature coronary artery disease and / or dyslipidemia who are refractory to statins or bile acid sequestrants. Furthermore, patients in whom elevated Lp(a) levels contribute to adverse clinical outcomes may benefit from Lp(a) screening, including those with a history of coronary artery bypass grafting, percutaneous transluminal coronary angioplasty, heart transplantation, and a family history of dyslipidemia.

[0007] In recent years, significant progress has been made in the treatment of dyslipidemia. Low-density lipoprotein cholesterol (LDL-C) is the primary intervention target for the prevention of atherosclerotic cardiovascular disease (ASCVD). However, many patients still have residual ASCVD risk after LDL-C-lowering therapy. Elevated lipoprotein a (Lp(a)) levels are associated with an increased risk of premature ASCVD. Interventions using statins, fibrates, or bile acid sequestrants have been ineffective in lowering Lp(a) levels. Newer lipid-lowering drugs, such as second-generation antisense oligonucleotides (ASOs), cholesteryl ester transfer protein (CETP) inhibitors, and proprotein convertase subtilisin-kexin 9 (PCSK9) inhibitors, can effectively lower Lp(a) levels.

[0008] Summary of the Invention

[0009] The purpose of the present disclosure is to provide an inhibitor targeting lipoprotein a (Lp(a)) with good efficacy, high safety and long-lasting efficacy.

[0010] The present disclosure provides dsRNA and methods of using the dsRNA to inhibit Lp(a) gene expression in cells or mammals, wherein the dsRNA targets the Lp(a) gene. Also provided herein are compositions and methods for treating pathological conditions and diseases in mammals caused by Lp(a) gene expression. The dsRNA directs the sequence-specific degradation of mRNA through a process known as RNA interference (RNAi).

[0011] In one aspect, the present disclosure provides an oligonucleotide or a pharmaceutically acceptable salt thereof for reducing lipoprotein a (Lp(a)) gene expression, the oligonucleotide comprising a sense strand and an antisense strand, the sense strand having a sequence having at least 80% sequence identity to a sequence shown in any one of SEQ ID NOs. 1-43, 45-66, 68-97, 99-206, 735-749, and 807-817, or a fragment thereof, or a modified sequence of the aforementioned sequence or fragment thereof, preferably a sequence having 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity; the antisense strand having a sequence having at least 80% sequence identity to a sequence shown in any one of SEQ ID NOs. 1-43, 45-66, 68-97, 99-206, 735-749, and 807-817, or a fragment thereof, or a modified sequence of the aforementioned sequence or fragment thereof; and the antisense strand having a sequence having at least 80% sequence identity to a sequence shown in any one of SEQ ID NOs. A sequence having at least 80% sequence identity to any one of the sequences shown in NO. 207-249, 251-276, 278-302, 304-404, 750-769 and 818-828 or a fragment thereof, or a modified sequence of the aforementioned sequence or a fragment thereof, preferably a sequence having 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity.

[0012] In another aspect, the present disclosure provides a conjugate for reducing lipoprotein a (Lp(a)) gene expression or a pharmaceutically acceptable salt thereof, comprising: (i) an oligonucleotide or a pharmaceutically acceptable salt thereof, and (ii) a ligand conjugated to the oligonucleotide or a pharmaceutically acceptable salt thereof, wherein at least one nucleotide of the oligonucleotide is conjugated to a targeting ligand.

[0013] In another aspect, the present disclosure provides a composition comprising the aforementioned oligonucleotide or a pharmaceutically acceptable salt thereof, or the aforementioned conjugate or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier.

[0014] In another aspect, the present disclosure provides use of the aforementioned oligonucleotide or a pharmaceutically acceptable salt thereof, or the aforementioned conjugate or a pharmaceutically acceptable salt thereof, or the aforementioned composition in the preparation of a medicament for treating and / or preventing Lp(a)-related diseases.

[0015] In another aspect, the present disclosure provides a method for treating and / or preventing Lp(a)-related disorders, conditions and / or pathologies in a subject by administering a therapeutic agent (e.g., the aforementioned oligonucleotide or a pharmaceutically acceptable salt thereof, or the aforementioned conjugate or a pharmaceutically acceptable salt thereof, or the aforementioned composition, or a vector or transgene encoding the oligonucleotide) to the subject.

[0016] In another aspect, the present disclosure provides methods for treating and / or preventing Lp(a)-related disorders, conditions and / or pathologies in a subject using the aforementioned oligonucleotides or pharmaceutically acceptable salts thereof, conjugates or pharmaceutically acceptable salts thereof, or compositions in combination with other drugs and / or other therapeutic methods.

[0017] Experiments have demonstrated that the candidate compounds disclosed herein can effectively reduce lipoprotein a (Lp(a)) levels in the body and are effective inhibitors of lipoprotein a (Lp(a)). The present disclosure provides a method for treating diseases or conditions associated with Lp(a) protein, comprising administering to a subject an effective dose of a dsRNA agent disclosed herein to inhibit Lp(a) gene expression. In certain aspects the Lp(a)-related disease or condition is a cardiovascular disease, wherein the cardiovascular disease comprises 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, hyperapolipoprotein betalipoproteinemia, cerebrovascular atherosclerosis, cerebrovascular disease, venous thrombosis, stroke, atherosclerosis, thrombosis, coronary heart disease or aortic stenosis and / or any other disease or pathology associated with elevated levels of Lp(a)-containing particles. In some embodiments, the method further comprises: administering to the subject an additional non-Lp(a) siRNA therapeutic agent in combination, wherein the non-Lp(a) siRNA therapeutic agent comprises one or more of the following, such as HMgCo-A reductase inhibitors (statins), ezetimibe, PCSK9 inhibitors, CTEP inhibitors, ANGPLT3 inhibitors, APOC3 inhibitors, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] FIG1 shows a map of the pFB-AAV-CAG-Gluc-2A-LPA-DS recombinant plasmid.

[0019] Figure 2 shows the efficacy of Lp(a)RNAi in Lp(a)-Gluc mice, wherein Figure 2A shows the efficacy of Lp(a)RNAi in one group of mice over 50 days, and Figure 2B shows the efficacy of Lp(a)RNAi in another group of mice over 36 days.

[0020] FIG3 shows the efficacy of Lp(a) siRNA in hLp(a) transgenic mice.

[0021] FIG4 shows the efficacy of Lp(a) siRNA in hLp(a) transgenic mice.

[0022] FIG5 shows the efficacy of Lp(a) siRNA in cynomolgus monkeys. DETAILED DESCRIPTION

[0023] In this disclosure, unless otherwise indicated, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Furthermore, terms and laboratory procedures related to protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, and immunology used herein are those widely used in the respective fields and are common procedures. To facilitate a better understanding of this disclosure, definitions and explanations of relevant terms are provided below.

[0024] As used herein, the term "approximately" or "approximately" as applied to one or more target values ​​refers to a value similar to the reference value. In certain embodiments, unless otherwise indicated or in addition apparent from context, the term "approximately" or "approximately" refers to a value falling within 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less of the reference value in either direction (greater than or less than) or less (unless such numerals will exceed 100% of possible values).

[0025] Described herein are Lp(a) (also known as apo(a)) RNA interference (RNAi) agents (also known as RNAi triggers or triggers) and compositions containing Lp(a) RNAi agents for selectively and effectively inhibiting the expression of the Lp(a) gene. Lp(a) is the name of the gene that encodes apolipoprotein(a) (apo(a)), a key component of lipoprotein(a) particles. The Lp(a) RNAi agents described herein can be used to prevent or treat the following diseases or to prepare a medicament for preventing or treating the following diseases, including but not limited to: Berger'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, cerebrovascular atherosclerosis, cerebrovascular disease, venous thrombosis, stroke, atherosclerosis, thrombosis, coronary heart disease or aortic valve stenosis and / or any other disease or pathology associated with elevated levels of Lp(a)-containing particles.

[0026] As used herein, "target sequence" refers to a continuous portion of the nucleotide sequence of an mRNA molecule formed during transcription of the Lp(a) gene, including mRNA that is a product of RNA processing of the primary transcript.

[0027] As used herein, term " complementary " refers to the structural relationship that allows nucleotide to form base pairs with each other between nucleotide (for example, on relative nucleic acid or on two nucleotides on the relative region of single nucleic acid chain).For example, the purine nucleotides complementary to the pyrimidine nucleotides of a nucleic acid can be base paired together by forming hydrogen bonds with each other.In some embodiments, complementary nucleotides can be base paired in Watson-Crick (Watson-Crick) mode or in any other manner that allows to form a stable duplex.In some embodiments, two nucleic acids can have and be complementary to each other to form the nucleotide sequence of complementary region, as described herein.

[0028] As used herein, the term "strand" refers to a single continuous sequence of nucleotides linked together by internucleotide bonds (e.g., phosphodiester bonds, phosphorothioate bonds). In some embodiments, the strand has two free ends, e.g., a 5'-end and a 3'-end.

[0029] As used herein, the term "deoxyribonucleotide" refers to a nucleotide that has a hydrogen at the 2' position of its pentose sugar compared to a ribonucleotide. A modified deoxyribonucleotide is a deoxyribonucleotide that has a modification or substitution of one or more atoms other than the 2' position, including a modification or substitution in or of a sugar, a phosphate group, or a base.

[0030] As used herein, the term "oligonucleotide" refers to a short nucleic acid, for example, a short nucleic acid less than 100 nucleotides in length. The oligonucleotide can comprise ribonucleotides, deoxyribonucleotides and / or modified nucleotides, including, for example, modified ribonucleotides. The oligonucleotide can be single-stranded or double-stranded. The oligonucleotide may or may not have a duplex region. As one group of non-limiting examples, the oligonucleotide can be, but is not limited to, small interfering RNA (siRNA), microRNA (miRNA), short hairpin RNA (shRNA), Dicer substrate interfering RNA (dsiRNA), antisense oligonucleotide, short siRNA or single-stranded siRNA. In some embodiments, the double-stranded oligonucleotide is an RNAi oligonucleotide.

[0031] As used herein, the term "double-stranded oligonucleotide" refers to an oligonucleotide that is substantially in duplex form. In some embodiments, the complementary base pairing of one or more duplex regions of a double-stranded oligonucleotide is formed between the antiparallel sequence of the nucleotides of the covalently separated nucleic acid chains. In some embodiments, the complementary base pairing of one or more duplex regions of a double-stranded oligonucleotide is formed between the antiparallel sequence of the nucleotides of the covalently attached nucleic acid chains. In some embodiments, the complementary base pairing of one or more duplex regions of a double-stranded oligonucleotide is formed from a single nucleic acid chain, and the single nucleic acid chain is folded (for example, via a hairpin) to provide the complementary antiparallel sequence of the nucleotides of base pairing together. In some embodiments, a double-stranded oligonucleotide comprises two covalently separated nucleic acid chains that are completely duplexed from each other. However, in some embodiments, a double-stranded oligonucleotide comprises partially duplexed, for example, two covalently separated nucleic acid chains with an overhang at one or both ends. In some embodiments, a double-stranded oligonucleotide comprises the antiparallel sequence of nucleotides, which are partially complementary, and therefore, can have one or more mispairings, and the mispairings can include internal mispairings or terminal mispairings.

[0032] As used herein, the terms "iRNA," "RNAi agent," "iRNA agent," and "RNA interference agent" are used interchangeably herein and refer to an agent that comprises RNA, as such terms are defined herein, and that mediates targeted cleavage of RNA transcripts via the RNA-induced silencing complex (RISC) pathway. RNA interference (RNAi) is a process that directs sequence-specific degradation of mRNA. RNAi modulates, for example, inhibits, the expression of Lp(a) in cells, for example, cells within an individual, such as a mammalian individual.

[0033] As used herein, "conjugation" refers to the covalent attachment of two or more chemical moieties, each with a specific function, to each other; accordingly, "conjugate" refers to a compound formed by covalent attachment of the chemical moieties. Furthermore, "siRNA conjugate" refers to a compound formed by covalent attachment of one or more chemical moieties with a specific function to siRNA. Hereinafter, the siRNA conjugates of the present disclosure will sometimes be referred to as "conjugates." siRNA conjugates should be understood as a general term for siRNA conjugates, including siRNA sense strand conjugates or siRNA antisense strand conjugates, depending on the context.

[0034] As used herein, the term "double-stranded RNA" or "dsRNA" refers to a complex of ribonucleic acid molecules having a duplex structure comprising two antiparallel and substantially complementary nucleic acid strands having "sense" and "antisense" orientations relative to the target RNA (i.e., the Lp(a) gene). In some embodiments of the present disclosure, double-stranded RNA (dsRNA) triggers degradation of the target RNA (e.g., mRNA) through a post-transcriptional gene silencing mechanism referred to herein as RNA interference or RNAi. Generally, the majority of the nucleotides in each strand of a dsRNA molecule are ribonucleotides, but as described in detail herein, each strand or both strands may also contain one or more non-ribonucleotides, such as deoxyribonucleotides or modified nucleotides. Additionally, as used herein, "iRNA" may contain ribonucleotides having chemical modifications; iRNA may contain substantial modifications at multiple nucleotides.

[0035] As used herein, the term "modified nucleotide" refers to a nucleotide that independently has a modified sugar moiety, a modified internucleotide linkage, or a modified nucleobase, or any combination thereof. Thus, the term "modified nucleotide" encompasses substitutions, additions, or removals of internucleoside linkages, sugar moieties, or nucleobases, such as functional groups or atoms. Modifications suitable for use with the agents of the present disclosure include all types of modifications disclosed herein or known in the art.

[0036] As used herein, the term "nucleotide overhang" refers to at least one unpaired nucleotide that protrudes from the duplex structure of a double-stranded iRNA. For example, a nucleotide overhang exists when the 3' end of one strand of a dsRNA extends beyond the 5' end of the other strand, or vice versa. A dsRNA may include an overhang of at least one nucleotide; alternatively, the overhang may include at least two nucleotides, at least three nucleotides, at least four nucleotides, at least five nucleotides or more. The nucleotide overhang may include or consist of nucleotide / nucleoside analogs, including deoxynucleotides / nucleosides. The overhang may be on the sense strand, the antisense strand, or any combination thereof. In addition, the nucleotides of the overhang may be present on the 5' end, the 3' end, or both ends of the antisense strand or the sense strand of the dsRNA.

[0037] As used herein, the term "naked sequence" refers to an unmodified nucleotide sequence.

[0038] As used herein, the term "inhibit" is used interchangeably with "knockdown," "reduction," "silencing," "downregulate," "suppression," and other similar terms, and includes any degree of inhibition.

[0039] The phrase "inhibiting the expression of Lp(a)" is intended to refer to inhibiting the expression of any Lp(a) gene (such as, for example, a mouse Lp(a) gene, a rat Lp(a) gene, a monkey Lp(a) gene, or a human Lp(a) gene), as well as variants or mutants of the Lp(a) gene. Thus, in the context of a genetically manipulated cell, cell population, or organism, the Lp(a) gene can be a wild-type Lp(a) gene, a mutant Lp(a) gene, or a transgenic Lp(a) gene.

[0040] "Inhibiting Lp(a) gene expression" includes any level of Lp(a) gene inhibition, for example, at least partial inhibition of Lp(a) gene expression. Lp(a) gene expression can be assessed based on the level or change in the level of any variable associated with Lp(a) gene expression, for example, mRNA level or Lp(a) protein level, or indirectly reflects inhibition of Lp(a)-based protein levels by inhibiting the mRNA level of Gluc and Lp(a) fusion protein genes, thereby inhibiting the level of Gluc protein.

[0041] The term "pharmaceutically acceptable salt" refers to salts that retain the biological effectiveness and characteristics of free alkali or free acid, which are not biologically or otherwise undesirable. These salts are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid (particularly hydrochloric acid) and organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methylsulfonic acid, ethylsulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcysteine. In addition, these salts can be prepared by adding inorganic bases or organic bases to the free acid. Salts derived from inorganic bases include but are not limited to alkali metal salts (such as sodium salts, potassium salts and lithium salts), ammonium salts, alkaline earth metal salts (such as calcium salts and magnesium salts). Salts derived from organic bases include, but are not limited to, salts formed with the following organic bases (e.g., organic amines): primary amines, secondary amines, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, and polyamine resins. The oligonucleotides of the present disclosure may also exist in the form of zwitterions. Particularly preferred pharmaceutically acceptable salts of the present disclosure are sodium salts, lithium salts, potassium salts, and trialkylammonium salts.

[0042] As used herein, the term "subject" refers to an animal that expresses the target gene endogenously or heterologously, such as a mammal, including primates (such as humans, non-human primates, such as monkeys and chimpanzees), non-primates (such as cows, pigs, horses, goats, rabbits, sheep, hamsters, guinea pigs, cats, dogs, rats or mice) or birds. In one embodiment, the subject is a human.

[0043] As used herein, the terms "treating" or "treatment" refer to a beneficial or desired result, such as reducing at least one sign or symptom of an Lp(a)-related disorder in a subject. Treatment also encompasses reducing one or more signs or symptoms associated with undesirable Lp(a) expression; lessening the extent of undesirable Lp(a) activation or stabilization; ameliorating or alleviating undesirable Lp(a) activation or stabilization. Treatment also encompasses reducing one or more signs or symptoms associated with undesirable Lp(a) expression. "Treatment" may also mean prolonging survival compared to expected survival in the absence of treatment.

[0044] As used herein, the terms "prevention" or "preventing" when used in reference to a disease or condition that would benefit from a reduction in Lp(a) gene expression or Lp(a) protein production.

[0045] As used herein, the term "therapeutically effective amount" is intended to encompass an amount of a RNAi agent that, when administered to a subject suffering from an Lp(a)-related disorder, is sufficient to affect treatment of the disease (e.g., by reducing, ameliorating, or maintaining an existing disease or one or more disease symptoms). The "therapeutically effective amount" may vary depending on the RNAi agent, how the agent is administered, the disease and its severity, as well as medical history, age, weight, family history, genetic makeup, type of previous or concomitant treatment (if any), and other individual characteristics of the subject to be treated.

[0046] As used herein, the term "prophylactically effective amount" is intended to encompass an amount of an RNAi agent that, when administered to a subject suffering from an Lp(a)-related disorder, is sufficient to prevent or ameliorate the disorder or one or more symptoms of the disorder. Amelioration of the disease includes slowing the progression of the disease or reducing the severity of the disease that later develops. A "prophylactically effective amount" may vary depending on the RNAi agent, how the agent is administered, the degree of disease risk, and the patient's medical history, age, weight, family history, genetic makeup, type of previous or concomitant therapy (if any), and other individual characteristics of the patient being treated.

[0047] In one aspect, the present disclosure provides an oligonucleotide or a pharmaceutically acceptable salt thereof for reducing lipoprotein a (Lp(a)) gene expression, the oligonucleotide comprising a sense chain and an antisense chain, the sense chain having a sequence having at least 80% sequence identity to a sequence shown in any one of SEQ ID NOs. 1-43, 45-66, 68-97, 99-206, 735-749 and 807-817 or a fragment thereof, or a modified sequence of a sequence or a fragment thereof, preferably a sequence having 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity; the antisense chain having a sequence having at least 80% sequence identity to a sequence shown in any one of SEQ ID NOs. 1-43, 45-66, 68-97, 99-206, 735-749 and 807-817 or a fragment thereof, or a modified sequence of a sequence or a fragment thereof; A sequence having at least 80% sequence identity to the sequence shown in any one of NO. 207-249, 251-276, 278-302, 304-404, 750-769 and 818-828 or a fragment thereof, or a modified sequence of a sequence or a fragment thereof, preferably a sequence having 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity.

[0048] In some embodiments, wherein each strand is independently 19 to 25 nucleotides in length.

[0049] In some embodiments, the antisense strand is 19 to 23 nucleotides in length.

[0050] In some embodiments, the sense strand is 19 to 23 nucleotides in length.

[0051] In some embodiments, the oligonucleotide comprises a 5' and / or 3'-overhang sequence having a length of one or more nucleotides, wherein the 5' and / or 3'-overhang sequence is present on the antisense strand and / or the sense strand. In one embodiment, the antisense strand of the oligonucleotide has 1 to 10 nucleotides at the 3' end or the 5' overhang at the end. For example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 nucleotides. In one embodiment, the sense strand of the dsRNA has 1 to 10 nucleotides at the 3' end or the 5' overhang at the end. For example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 nucleotides. In another embodiment, the one or more nucleotides in the overhang are replaced by nucleoside thiophosphates.

[0052] In some embodiments, the antisense strand has one or two overhangs.

[0053] In some embodiments, the sense strand has one or two overhangs.

[0054] In some embodiments, the oligonucleotide comprises a 3'-overhang sequence that is 1 or 2 nucleotides in length.

[0055] In some embodiments, the oligonucleotide comprises a 5'-overhang sequence that is 1 or 2 nucleotides in length.

[0056] In some embodiments, the 3'-overhang sequence is present on the antisense strand. In some embodiments, the overhang sequence is selected from the group consisting of: AA, AG, AU, C, CA, CC, CG, CU, G, GA, GC, GG, GU, U, UA, UC, UG, UU.

[0057] In some embodiments, the 5'-overhang sequence is present on the antisense strand.In some embodiments, the overhang sequence is selected from the group consisting of: A and G.

[0058] In some embodiments, the oligonucleotide comprises an antisense strand and a sense strand each ranging from 19 to 23 nucleotides in length.

[0059] In some embodiments, the sense strand and the antisense strand form a duplex region.

[0060] In some embodiments, the sense strand and the antisense strand are respectively in a 19 / 21 paired, 21 / 21 paired, 21 / 23 paired, or 23 / 23 paired duplex structure.

[0061] In some embodiments, the oligonucleotide comprises a 5' overhang of 1 nucleotide in length and a 3'-overhang sequence, wherein the 5' overhang and the 3'-overhang sequence are present on the antisense strand, and wherein the sense strand is 19 nucleotides in length and the antisense strand is 21 nucleotides in length, such that the sense strand and the antisense strand form a duplex of 19 nucleotides in length.

[0062] In some embodiments, the oligonucleotide comprises a 3'-overhang sequence that is 2 nucleotides in length, wherein the 3'-overhang sequence is present on the antisense strand, and wherein the sense strand is 19 nucleotides in length and the antisense strand is 21 nucleotides in length, such that the sense strand and the antisense strand form a duplex that is 19 nucleotides in length.

[0063] In some embodiments, the oligonucleotide comprises a 3'-overhang sequence that is 2 nucleotides in length, wherein the 3'-overhang sequence is present on the antisense strand and the sense strand, and wherein the sense strand is 21 nucleotides in length and the antisense strand is 21 nucleotides in length, such that the sense strand and the antisense strand form a duplex that is 19 nucleotides in length.

[0064] In some embodiments, the oligonucleotide comprises a 3'-overhang sequence that is 2 nucleotides in length, wherein the 3'-overhang sequence is present on the antisense strand, and wherein the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length, such that the sense strand and the antisense strand form a duplex that is 21 nucleotides in length.

[0065] In some embodiments, the oligonucleotide comprises a 3'-overhang sequence that is 2 nucleotides in length, wherein the 3'-overhang sequence is present on the antisense strand and the sense strand, and wherein the sense strand is 23 nucleotides in length and the antisense strand is 23 nucleotides in length, such that the sense strand and the antisense strand form a duplex that is 21 nucleotides in length.

[0066] In some embodiments, the pharmaceutically acceptable salt of the oligonucleotide can be prepared by adding an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include but are not limited to alkali metal salts (such as sodium salts, potassium salts and lithium salts), ammonium salts, alkaline earth metal salts (such as calcium salts and magnesium salts). Salts derived from organic bases (such as organic amines) include but are not limited to salts formed with the following organic bases: primary amines, secondary amines and tertiary amines, substituted amines include naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, polyamine resins.

[0067] In some embodiments, examples of pharmaceutically acceptable salts of oligonucleotides include, but are not limited to, ammonium salts, such as salts of tertiary alkylamine compounds (eg, triethylamine salts), metal salts such as sodium salts, potassium salts, and magnesium salts.

[0068] In some embodiments, the oligonucleotide or its salt may be in the form of a hydrate or a solvate.

[0069] In some embodiments, the oligonucleotide comprises at least one modified nucleotide.

[0070] In some embodiments, the oligonucleotide comprises at least one 2'-modified nucleotide.

[0071] In some embodiments, the 2'-modified nucleotides are selected from one or more of 2'-alkoxy modified nucleotides, 2'-substituted alkoxy modified nucleotides, 2'-alkyl modified nucleotides, 2'-substituted alkyl modified nucleotides, 2'-amino modified nucleotides, 2'-substituted amino modified nucleotides, 2'-fluoro modified nucleotides, and 2'-deoxy nucleotides.

[0072] In some embodiments, the 2'-modification is a modification selected from the group consisting of 2'-methoxy, 2'-acetamido, 2'-aminoethyl, 2'-fluoro, 2'-O-methoxyethyl.

[0073] In some embodiments, the oligonucleotide comprises a diol nucleic acid (GNA) modification, and the ribonucleotide has a diol nucleic acid structure as shown in formula (I), preferably an S-isomer. In some embodiments, the oligonucleotide comprises Tgn, which is a thymidine-diol nucleic acid as shown in formula (II), preferably an S-isomer thereof. In some embodiments, the oligonucleotide comprises Cgn, which is a cytidine-diol nucleic acid as shown in formula (III), preferably an S-isomer thereof. In some embodiments, the oligonucleotide comprises Agn, which is an adenosine-diol nucleic acid as shown in formula (IV), preferably an S-isomer thereof. In some embodiments, the oligonucleotide comprises Ggn, which is a guanosine-diol nucleic acid as shown in formula (V), preferably an S-isomer thereof;

[0074] In some embodiments, the oligonucleotide comprises a nucleoside-2'-phosphate of formula (VI), wherein Base represents a nucleic acid base selected from A, G, C, and U. In some embodiments, the 5'-phosphate analog modified nucleotide has a vinylphosphonate modified nucleotide of formula (XII). In some embodiments. In some embodiments, the oligonucleotide comprises U-2'5', which is a uridine-2'-phosphate of formula (VII). In some embodiments, the oligonucleotide comprises C-2'5', which is a cytidine-2'-phosphate of formula (VIII). In some embodiments, the oligonucleotide comprises A-2'5', which is an adenosine-2'-phosphate of formula (IX). In some embodiments, the oligonucleotide comprises G-2'5', which is a guanosine-2'-phosphate of formula (X);

[0075] In some embodiments, the oligonucleotide has a 5'-phosphate analog modified nucleotide at the 5' end. In some embodiments, the 5'-phosphate analog modified nucleotide has a vinyl phosphonate modified nucleotide shown in formula (XI), wherein R is selected from H, fluorine, 2'-methoxy, 2'-acetylamino, 2'-aminoethyl and 2'-O-methoxyethyl; Base represents a nucleic acid base selected from A, G, C, T and U. In some embodiments, the 5'-phosphate analog modified nucleotide has a vinyl phosphonate modified nucleotide shown in formula (XII). In some embodiments, the oligonucleotide has APU at the 5' end, which is a 5'-phosphate analog modified uridylic acid (2'-acetylamino-5'-vinylphosphonate-uridylic acid) shown in formula (XIII). In some embodiments, the oligonucleotide has VPU at the 5' end, which is a 5'-phosphate analog modified uridylic acid (2'-methoxy-5'-vinylphosphonate-uridylic acid) shown in formula (XIV);

[0076] In some embodiments, the oligonucleotide comprises an inverted ribonucleotide (3'-3' linked nucleotide) as shown in formula (XV). In some embodiments, the oligonucleotide comprises invdA, which is an inverted deoxyadenylic acid as shown in formula (XVI);

[0077] In some embodiments, the oligonucleotide comprises formula M, which is 2'-O-methyl-6-(3-(2-carboxyethyl)phenyl)-adenylic acid shown in formula (XVII);

[0078] In some embodiments, the oligonucleotide comprises at least one modified internucleotide linkage.

[0079] In some embodiments, at least one modified internucleotide bond is a phosphorothioate bond. The phosphorothioate internucleotide bond modification can occur on any nucleotide of the sense strand, antisense strand, or both strands at any position in the chain. For example, the internucleotide bond modification can occur on each nucleotide on the sense strand or antisense strand; each internucleotide bond modification can occur in an alternating pattern on the sense strand or antisense strand; or the sense strand or antisense strand can contain two internucleotide bond modifications in an alternating pattern. The alternating pattern of the internucleotide bond modification on the sense strand can be the same or different from the antisense strand, and the alternating pattern of the internucleotide bond modification on the sense strand can have an offset relative to the alternating pattern of the internucleotide bond on the antisense strand. In one embodiment, the double-stranded RNAi agent includes 4 to 8 phosphorothioate internucleotide bonds. In some embodiments, the antisense strand includes two phosphorothioate internucleotide bonds at the 5' end and two phosphorothioate internucleotide bonds at the 3' end, and the sense strand includes at least two phosphorothioate internucleotide bonds at the 5' end or the 3' end.

[0080] In some embodiments, the sense strand is selected from the group consisting of SEQ ID NO. 8, 9, 38, 39, 40, 41, 55, 63, 64, 65, 66, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 110 08, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 1 48, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 1 88, 189, 190, 191, 192, 193, 194, 195, 196, 197, 735, 736, 737, 738, 739, 740, 741, 742, 743, 744, 745, 746, 747, 748, 749, 807, 808, 809, 810, 811, 812, 813, 814, 815, 816, 817, or any one of the modified oligonucleotides of SEQ ID NO. 405-419, 421-451, 453-556, 770-784, 829-839; the antisense strand is selected from SEQ ID NO. NO. 258, 259, 265, 215, 273, 274, 275, 276, 278, 281, 283, 284, 286, 287, 297, 304, 306, 309, 316, 320, 326, 388, 389, 404, 750, 751, 752, 753, 754, 755, 756, 757, 758, 759, 760, 761, 762, 763, 764, 765, 766, 767, 768, 769, 818, 819, 820, 821, 822, 823, 824, 825, 826, 827, 828, an unmodified oligonucleotide of any one of SEQ ID NO.The modified oligonucleotide according to any one of 557-572, 574-598, 600-734, and 785-806.

[0081] In some embodiments, the oligonucleotide is selected from any of the following sense and antisense strand combinations:

[0082] (1) the sense strand comprises the sequence shown in SEQ ID NO. 9, and the antisense strand comprises the sequence shown in SEQ ID NO. 215;

[0083] (2) the sense strand comprises the sequence shown in SEQ ID NO. 38, and the antisense strand comprises the sequence shown in SEQ ID NO. 258;

[0084] (3) the sense strand comprises the sequence shown in SEQ ID NO. 40, and the antisense strand comprises the sequence shown in SEQ ID NO. 259;

[0085] (4) the sense strand comprises the sequence shown in SEQ ID NO. 55, and the antisense strand comprises the sequence shown in SEQ ID NO. 265;

[0086] (5) the sense strand comprises the sequence shown in SEQ ID NO. 63, and the antisense strand comprises the sequence shown in SEQ ID NO. 273;

[0087] (6) the sense strand comprises the sequence shown in SEQ ID NO. 64, and the antisense strand comprises the sequence shown in SEQ ID NO. 274;

[0088] (7) the sense strand comprises the sequence shown in SEQ ID NO. 65, and the antisense strand comprises the sequence shown in SEQ ID NO. 275;

[0089] (8) the sense strand comprises the sequence shown in SEQ ID NO. 66, and the antisense strand comprises the sequence shown in SEQ ID NO. 276;

[0090] (9) the sense strand comprises the sequence shown in SEQ ID NO. 68, and the antisense strand comprises the sequence shown in SEQ ID NO. 274;

[0091] (10) the sense strand comprises the sequence shown in SEQ ID NO. 74, and the antisense strand comprises the sequence shown in SEQ ID NO. 278;

[0092] (11) the sense strand comprises the sequence shown in SEQ ID NO. 77, and the antisense strand comprises the sequence shown in SEQ ID NO. 281;

[0093] (12) the sense strand comprises the sequence shown in SEQ ID NO. 79, and the antisense strand comprises the sequence shown in SEQ ID NO. 283;

[0094] (13) the sense strand comprises the sequence shown in SEQ ID NO. 80, and the antisense strand comprises the sequence shown in SEQ ID NO. 284;

[0095] (14) the sense strand comprises the sequence shown in SEQ ID NO. 80, and the antisense strand comprises the sequence shown in SEQ ID NO. 767;

[0096] (15) the sense strand comprises the sequence shown in SEQ ID NO. 80, and the antisense strand comprises the sequence shown in SEQ ID NO. 768;

[0097] (16) the sense strand comprises the sequence shown in SEQ ID NO. 80, and the antisense strand comprises the sequence shown in SEQ ID NO. 769;

[0098] (17) the sense strand comprises the sequence shown in SEQ ID NO. 82, and the antisense strand comprises the sequence shown in SEQ ID NO. 286;

[0099] (18) the sense strand comprises the sequence shown in SEQ ID NO. 83, and the antisense strand comprises the sequence shown in SEQ ID NO. 287;

[0100] (19) the sense strand comprises the sequence shown in SEQ ID NO. 93, and the antisense strand comprises the sequence shown in SEQ ID NO. 297;

[0101] (20) the sense strand comprises the sequence shown in SEQ ID NO. 99, and the antisense strand comprises the sequence shown in SEQ ID NO. 304;

[0102] (21) the sense strand comprises the sequence shown in SEQ ID NO. 101, and the antisense strand comprises the sequence shown in SEQ ID NO. 306;

[0103] (22) the sense strand comprises the sequence shown in SEQ ID NO. 104, and the antisense strand comprises the sequence shown in SEQ ID NO. 309;

[0104] (23) the sense strand comprises the sequence shown in SEQ ID NO. 111, and the antisense strand comprises the sequence shown in SEQ ID NO. 316;

[0105] (24) the sense strand comprises the sequence shown in SEQ ID NO. 115, and the antisense strand comprises the sequence shown in SEQ ID NO. 320;

[0106] (25) the sense strand comprises the sequence shown in SEQ ID NO. 121, and the antisense strand comprises the sequence shown in SEQ ID NO. 326;

[0107] (26) the sense strand comprises the sequence shown in SEQ ID NO. 182, and the antisense strand comprises the sequence shown in SEQ ID NO. 388;

[0108] (27) the sense strand comprises the sequence shown in SEQ ID NO. 183, and the antisense strand comprises the sequence shown in SEQ ID NO. 389;

[0109] (28) the sense strand comprises the sequence shown in SEQ ID NO. 197, and the antisense strand comprises the sequence shown in SEQ ID NO. 404;

[0110] (29) the sense strand comprises the sequence shown in SEQ ID NO. 197, and the antisense strand comprises the sequence shown in SEQ ID NO. 765;

[0111] (30) the sense strand comprises the sequence shown in SEQ ID NO. 197, and the antisense strand comprises the sequence shown in SEQ ID NO. 766;

[0112] (31) the sense strand comprises the sequence shown in SEQ ID NO. 735, and the antisense strand comprises the sequence shown in SEQ ID NO. 750;

[0113] (32) the sense strand comprises the sequence shown in SEQ ID NO. 736, and the antisense strand comprises the sequence shown in SEQ ID NO. 751;

[0114] (33) the sense strand comprises the sequence shown in SEQ ID NO. 737, and the antisense strand comprises the sequence shown in SEQ ID NO. 752;

[0115] (34) the sense strand comprises the sequence shown in SEQ ID NO. 738, and the antisense strand comprises the sequence shown in SEQ ID NO. 753;

[0116] (35) the sense strand comprises the sequence shown in SEQ ID NO. 739, and the antisense strand comprises the sequence shown in SEQ ID NO. 754;

[0117] (36) the sense strand comprises the sequence shown in SEQ ID NO. 740, and the antisense strand comprises the sequence shown in SEQ ID NO. 755;

[0118] (37) the sense strand comprises the sequence shown in SEQ ID NO. 741, and the antisense strand comprises the sequence shown in SEQ ID NO. 756;

[0119] (38) the sense strand comprises the sequence shown in SEQ ID NO. 742, and the antisense strand comprises the sequence shown in SEQ ID NO. 757;

[0120] (39) the sense strand comprises the sequence shown in SEQ ID NO. 743, and the antisense strand comprises the sequence shown in SEQ ID NO. 758;

[0121] (40) the sense strand comprises the sequence shown in SEQ ID NO. 744, and the antisense strand comprises the sequence shown in SEQ ID NO. 759;

[0122] (41) the sense strand comprises the sequence shown in SEQ ID NO. 745, and the antisense strand comprises the sequence shown in SEQ ID NO. 760;

[0123] (42) the sense strand comprises the sequence shown in SEQ ID NO. 746, and the antisense strand comprises the sequence shown in SEQ ID NO. 761;

[0124] (43) the sense strand comprises the sequence shown in SEQ ID NO. 747, and the antisense strand comprises the sequence shown in SEQ ID NO. 762;

[0125] (44) the sense strand comprises the sequence shown in SEQ ID NO. 748, and the antisense strand comprises the sequence shown in SEQ ID NO. 763;

[0126] (45) the sense strand comprises the sequence shown in SEQ ID NO. 749, and the antisense strand comprises the sequence shown in SEQ ID NO. 764;

[0127] (46) the sense strand comprises the sequence shown in SEQ ID NO. 807, and the antisense strand comprises the sequence shown in SEQ ID NO. 818;

[0128] (47) the sense strand comprises the sequence shown in SEQ ID NO. 808, and the antisense strand comprises the sequence shown in SEQ ID NO. 819;

[0129] (48) the sense strand comprises the sequence shown in SEQ ID NO. 809, and the antisense strand comprises the sequence shown in SEQ ID NO. 820;

[0130] (49) the sense strand comprises the sequence shown in SEQ ID NO. 810, and the antisense strand comprises the sequence shown in SEQ ID NO. 821;

[0131] (50) the sense strand comprises the sequence shown in SEQ ID NO. 811, and the antisense strand comprises the sequence shown in SEQ ID NO. 822;

[0132] (51) the sense strand comprises the sequence shown in SEQ ID NO. 812, and the antisense strand comprises the sequence shown in SEQ ID NO. 823;

[0133] (52) the sense strand comprises the sequence shown in SEQ ID NO. 813, and the antisense strand comprises the sequence shown in SEQ ID NO. 824;

[0134] (53) the sense strand comprises the sequence shown in SEQ ID NO. 814, and the antisense strand comprises the sequence shown in SEQ ID NO. 825;

[0135] (54) the sense strand comprises the sequence shown in SEQ ID NO. 815, and the antisense strand comprises the sequence shown in SEQ ID NO. 826;

[0136] (55) the sense strand comprises the sequence shown in SEQ ID NO. 816, and the antisense strand comprises the sequence shown in SEQ ID NO. 827; and

[0137] (56) the sense strand comprises the sequence shown in SEQ ID NO. 817, and the antisense strand comprises the sequence shown in SEQ ID NO. 828;

[0138] wherein each strand is independently 19 to 25 nucleotides in length.

[0139] In some embodiments, the oligonucleotide comprises any one selected from the following sense and antisense strand combinations:

[0140] (1) the sense strand comprises the sequence shown in SEQ ID NO. 405, and the antisense strand comprises the sequence shown in SEQ ID NO. 557;

[0141] (2) the sense strand comprises the sequence shown in SEQ ID NO. 406, and the antisense strand comprises the sequence shown in SEQ ID NO. 558;

[0142] (3) the sense strand comprises the sequence shown in SEQ ID NO. 407, and the antisense strand comprises the sequence shown in SEQ ID NO. 559;

[0143] (4) the sense strand comprises the sequence shown in SEQ ID NO. 408, and the antisense strand comprises the sequence shown in SEQ ID NO. 560;

[0144] (5) the sense strand comprises the sequence shown in SEQ ID NO. 409, and the antisense strand comprises the sequence shown in SEQ ID NO. 561;

[0145] (6) the sense strand comprises the sequence shown in SEQ ID NO. 410, and the antisense strand comprises the sequence shown in SEQ ID NO. 562;

[0146] (7) the sense strand comprises the sequence shown in SEQ ID NO. 411, and the antisense strand comprises the sequence shown in SEQ ID NO. 563;

[0147] (8) the sense strand comprises the sequence shown in SEQ ID NO. 412, and the antisense strand comprises the sequence shown in SEQ ID NO. 564;

[0148] (9) the sense strand comprises the sequence shown in SEQ ID NO. 413, and the antisense strand comprises the sequence shown in SEQ ID NO. 565;

[0149] (10) the sense strand comprises the sequence shown in SEQ ID NO. 414, and the antisense strand comprises the sequence shown in SEQ ID NO. 566;

[0150] (11) the sense strand comprises the sequence shown in SEQ ID NO. 415, and the antisense strand comprises the sequence shown in SEQ ID NO. 567;

[0151] (12) the sense strand comprises the sequence shown in SEQ ID NO. 416, and the antisense strand comprises the sequence shown in SEQ ID NO. 568;

[0152] (13) the sense strand comprises the sequence shown in SEQ ID NO. 417, and the antisense strand comprises the sequence shown in SEQ ID NO. 569;

[0153] (14) the sense strand comprises the sequence shown in SEQ ID NO. 418, and the antisense strand comprises the sequence shown in SEQ ID NO. 570;

[0154] (15) the sense strand comprises the sequence shown in SEQ ID NO. 419, and the antisense strand comprises the sequence shown in SEQ ID NO. 562;

[0155] (16) the sense strand comprises the sequence shown in SEQ ID NO. 419, and the antisense strand comprises the sequence shown in SEQ ID NO. 571;

[0156] (17) the sense strand comprises the sequence shown in SEQ ID NO. 410, and the antisense strand comprises the sequence shown in SEQ ID NO. 572;

[0157] (18) the sense strand comprises the sequence shown in SEQ ID NO. 421, and the antisense strand comprises the sequence shown in SEQ ID NO. 562;

[0158] (19) the sense strand comprises the sequence shown in SEQ ID NO. 427, and the antisense strand comprises the sequence shown in SEQ ID NO. 574;

[0159] (20) the sense strand comprises the sequence shown in SEQ ID NO. 430, and the antisense strand comprises the sequence shown in SEQ ID NO. 577;

[0160] (21) the sense strand comprises the sequence shown in SEQ ID NO. 432, and the antisense strand comprises the sequence shown in SEQ ID NO. 579;

[0161] (22) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 580;

[0162] (23) the sense strand comprises the sequence shown in SEQ ID NO. 435, and the antisense strand comprises the sequence shown in SEQ ID NO. 582;

[0163] (24) the sense strand comprises the sequence shown in SEQ ID NO. 436, and the antisense strand comprises the sequence shown in SEQ ID NO. 583;

[0164] (25) the sense strand comprises the sequence shown in SEQ ID NO. 446, and the antisense strand comprises the sequence shown in SEQ ID NO. 593;

[0165] (26) the sense strand comprises the sequence shown in SEQ ID NO. 453, and the antisense strand comprises the sequence shown in SEQ ID NO. 600;

[0166] (27) the sense strand comprises the sequence shown in SEQ ID NO. 455, and the antisense strand comprises the sequence shown in SEQ ID NO. 602;

[0167] (28) the sense strand comprises the sequence shown in SEQ ID NO. 458, and the antisense strand comprises the sequence shown in SEQ ID NO. 605;

[0168] (29) the sense strand comprises the sequence shown in SEQ ID NO. 465, and the antisense strand comprises the sequence shown in SEQ ID NO. 612;

[0169] (30) the sense strand comprises the sequence shown in SEQ ID NO. 469, and the antisense strand comprises the sequence shown in SEQ ID NO. 616;

[0170] (31) the sense strand comprises the sequence shown in SEQ ID NO. 475, and the antisense strand comprises the sequence shown in SEQ ID NO. 622;

[0171] (32) the sense strand comprises the sequence shown in SEQ ID NO. 537, and the antisense strand comprises the sequence shown in SEQ ID NO. 685;

[0172] (33) the sense strand comprises the sequence shown in SEQ ID NO. 538, and the antisense strand comprises the sequence shown in SEQ ID NO. 686;

[0173] (34) the sense strand comprises the sequence shown in SEQ ID NO. 419, and the antisense strand comprises the sequence shown in SEQ ID NO. 700;

[0174] (35) the sense strand comprises the sequence shown in SEQ ID NO. 432, and the antisense strand comprises the sequence shown in SEQ ID NO. 701;

[0175] (36) the sense strand comprises the sequence shown in SEQ ID NO. 419, and the antisense strand comprises the sequence shown in SEQ ID NO. 702;

[0176] (37) the sense strand comprises the sequence shown in SEQ ID NO. 430, and the antisense strand comprises the sequence shown in SEQ ID NO. 703;

[0177] (38) the sense strand comprises the sequence shown in SEQ ID NO. 432, and the antisense strand comprises the sequence shown in SEQ ID NO. 704;

[0178] (39) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 705;

[0179] (40) the sense strand comprises the sequence shown in SEQ ID NO. 435, and the antisense strand comprises the sequence shown in SEQ ID NO. 706;

[0180] (41) the sense strand comprises the sequence shown in SEQ ID NO. 432, and the antisense strand comprises the sequence shown in SEQ ID NO. 707;

[0181] (42) the sense strand comprises the sequence shown in SEQ ID NO. 553, and the antisense strand comprises the sequence shown in SEQ ID NO. 710;

[0182] (43) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 712;

[0183] (44) the sense strand comprises the sequence shown in SEQ ID NO. 553, and the antisense strand comprises the sequence shown in SEQ ID NO. 716;

[0184] (45) the sense strand comprises the sequence shown in SEQ ID NO. 553, and the antisense strand comprises the sequence shown in SEQ ID NO. 717;

[0185] (46) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 725;

[0186] (47) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 726;

[0187] (48) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 727;

[0188] (49) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 729;

[0189] (50) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 733;

[0190] (51) the sense strand comprises the sequence shown in SEQ ID NO. 553, and the antisense strand comprises the sequence shown in SEQ ID NO. 785;

[0191] (52) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 786;

[0192] (53) the sense strand comprises the sequence shown in SEQ ID NO. 770, and the antisense strand comprises the sequence shown in SEQ ID NO. 787;

[0193] (54) the sense strand comprises the sequence shown in SEQ ID NO. 771, and the antisense strand comprises the sequence shown in SEQ ID NO. 788;

[0194] (55) the sense strand comprises the sequence shown in SEQ ID NO. 772, and the antisense strand comprises the sequence shown in SEQ ID NO. 789;

[0195] (56) the sense strand comprises the sequence shown in SEQ ID NO. 773, and the antisense strand comprises the sequence shown in SEQ ID NO. 790;

[0196] (57) the sense strand comprises the sequence shown in SEQ ID NO. 774, and the antisense strand comprises the sequence shown in SEQ ID NO. 791;

[0197] (58) the sense strand comprises the sequence shown in SEQ ID NO. 775, and the antisense strand comprises the sequence shown in SEQ ID NO. 792;

[0198] (59) the sense strand comprises the sequence shown in SEQ ID NO. 776, and the antisense strand comprises the sequence shown in SEQ ID NO. 793;

[0199] (60) the sense strand comprises the sequence shown in SEQ ID NO. 777, and the antisense strand comprises the sequence shown in SEQ ID NO. 794;

[0200] (61) the sense strand comprises the sequence shown in SEQ ID NO. 778, and the antisense strand comprises the sequence shown in SEQ ID NO. 795;

[0201] (62) the sense strand comprises the sequence shown in SEQ ID NO. 779, and the antisense strand comprises the sequence shown in SEQ ID NO. 796;

[0202] (63) the sense strand comprises the sequence shown in SEQ ID NO. 780, and the antisense strand comprises the sequence shown in SEQ ID NO. 797;

[0203] (64) the sense strand comprises the sequence shown in SEQ ID NO. 781, and the antisense strand comprises the sequence shown in SEQ ID NO. 798;

[0204] (65) the sense strand comprises the sequence shown in SEQ ID NO. 782, and the antisense strand comprises the sequence shown in SEQ ID NO. 799;

[0205] (66) the sense strand comprises the sequence shown in SEQ ID NO. 783, and the antisense strand comprises the sequence shown in SEQ ID NO. 800;

[0206] (67) the sense strand comprises the sequence shown in SEQ ID NO. 784, and the antisense strand comprises the sequence shown in SEQ ID NO. 801;

[0207] (68) the sense strand comprises the sequence shown in SEQ ID NO. 553, and the antisense strand comprises the sequence shown in SEQ ID NO. 802;

[0208] (69) the sense strand comprises the sequence shown in SEQ ID NO. 553, and the antisense strand comprises the sequence shown in SEQ ID NO. 803;

[0209] (70) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 804;

[0210] (71) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 805;

[0211] (72) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 806;

[0212] (73) the sense strand comprises the sequence shown in SEQ ID NO. 772, and the antisense strand comprises the sequence shown in SEQ ID NO. 841;

[0213] (74) the sense strand comprises the sequence shown in SEQ ID NO. 775, and the antisense strand comprises the sequence shown in SEQ ID NO. 842;

[0214] (75) the sense strand comprises the sequence shown in SEQ ID NO. 776, and the antisense strand comprises the sequence shown in SEQ ID NO. 843;

[0215] (76) the sense strand comprises the sequence shown in SEQ ID NO. 777, and the antisense strand comprises the sequence shown in SEQ ID NO. 844;

[0216] (77) the sense strand comprises the sequence shown in SEQ ID NO. 783, and the antisense strand comprises the sequence shown in SEQ ID NO. 848;

[0217] (78) the sense strand comprises the sequence shown in SEQ ID NO. 783, and the antisense strand comprises the sequence shown in SEQ ID NO. 854;

[0218] (79) the sense strand comprises the sequence shown in SEQ ID NO. 784, and the antisense strand comprises the sequence shown in SEQ ID NO. 849;

[0219] (80) the sense strand comprises the sequence shown in SEQ ID NO. 831, and the antisense strand comprises the sequence shown in SEQ ID NO. 858;

[0220] (81) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 869;

[0221] (82) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 840;

[0222] (83) the sense strand comprises the sequence shown in SEQ ID NO. 772, and the antisense strand comprises the sequence shown in SEQ ID NO. 841;

[0223] (84) the sense strand comprises the sequence shown in SEQ ID NO. 775, and the antisense strand comprises the sequence shown in SEQ ID NO. 842;

[0224] (85) the sense strand comprises the sequence shown in SEQ ID NO. 776, and the antisense strand comprises the sequence shown in SEQ ID NO. 843;

[0225] (86) the sense strand comprises the sequence shown in SEQ ID NO. 777, and the antisense strand comprises the sequence shown in SEQ ID NO. 844;

[0226] (87) the sense strand comprises the sequence shown in SEQ ID NO. 783, and the antisense strand comprises the sequence shown in SEQ ID NO. 848;

[0227] (88) the sense strand comprises the sequence shown in SEQ ID NO. 783, and the antisense strand comprises the sequence shown in SEQ ID NO. 854;

[0228] (89) the sense strand comprises the sequence shown in SEQ ID NO. 784, and the antisense strand comprises the sequence shown in SEQ ID NO. 849;

[0229] (90) the sense strand comprises the sequence shown in SEQ ID NO. 831, and the antisense strand comprises the sequence shown in SEQ ID NO. 858; and

[0230] (91) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 869;

[0231] wherein each strand is independently 19 to 25 nucleotides in length.

[0232] In some embodiments, the oligonucleotide comprises any one selected from the following sense and antisense strand combinations:

[0233] (1) the sense strand comprises the sequence shown in SEQ ID NO. 406, and the antisense strand comprises the sequence shown in SEQ ID NO. 558;

[0234] (2) the sense strand comprises the sequence shown in SEQ ID NO. 407, and the antisense strand comprises the sequence shown in SEQ ID NO. 559;

[0235] (3) the sense strand comprises the sequence shown in SEQ ID NO. 409, and the antisense strand comprises the sequence shown in SEQ ID NO. 561;

[0236] (4) the sense strand comprises the sequence shown in SEQ ID NO. 410, and the antisense strand comprises the sequence shown in SEQ ID NO. 562;

[0237] (5) the sense strand comprises the sequence shown in SEQ ID NO. 411, and the antisense strand comprises the sequence shown in SEQ ID NO. 563;

[0238] (6) the sense strand comprises the sequence shown in SEQ ID NO. 419, and the antisense strand comprises the sequence shown in SEQ ID NO. 702;

[0239] (7) the sense strand comprises the sequence shown in SEQ ID NO. 430, and the antisense strand comprises the sequence shown in SEQ ID NO. 703;

[0240] (8) the sense strand comprises the sequence shown in SEQ ID NO. 432, and the antisense strand comprises the sequence shown in SEQ ID NO. 704;

[0241] (9) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 705;

[0242] (10) the sense strand comprises the sequence shown in SEQ ID NO. 435, and the antisense strand comprises the sequence shown in SEQ ID NO. 706;

[0243] (11) the sense strand comprises the sequence shown in SEQ ID NO. 432, and the antisense strand comprises the sequence shown in SEQ ID NO. 707;

[0244] (12) the sense strand comprises the sequence shown in SEQ ID NO. 553, and the antisense strand comprises the sequence shown in SEQ ID NO. 710;

[0245] (13) the sense strand comprises the sequence shown in SEQ ID NO. 553, and the antisense strand comprises the sequence shown in SEQ ID NO. 716;

[0246] (14) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 712;

[0247] (15) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 727;

[0248] (16) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 729;

[0249] (17) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 733;

[0250] (18) the sense strand comprises the sequence shown in SEQ ID NO. 553, and the antisense strand comprises the sequence shown in SEQ ID NO. 785;

[0251] (19) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 786;

[0252] (20) the sense strand comprises the sequence shown in SEQ ID NO. 453, and the antisense strand comprises the sequence shown in SEQ ID NO. 600;

[0253] (21) the sense strand comprises the sequence shown in SEQ ID NO. 537, and the antisense strand comprises the sequence shown in SEQ ID NO. 685;

[0254] (22) the sense strand comprises the sequence shown in SEQ ID NO. 538, and the antisense strand comprises the sequence shown in SEQ ID NO. 686;

[0255] (23) the sense strand comprises the sequence shown in SEQ ID NO. 433, and the antisense strand comprises the sequence shown in SEQ ID NO. 840;

[0256] (24) the sense strand comprises the sequence shown in SEQ ID NO. 772, and the antisense strand comprises the sequence shown in SEQ ID NO. 841;

[0257] (25) the sense strand comprises the sequence shown in SEQ ID NO. 775, and the antisense strand comprises the sequence shown in SEQ ID NO. 842;

[0258] (26) the sense strand comprises the sequence shown in SEQ ID NO. 776, and the antisense strand comprises the sequence shown in SEQ ID NO. 843;

[0259] (27) the sense strand comprises the sequence shown in SEQ ID NO. 777, and the antisense strand comprises the sequence shown in SEQ ID NO. 844;

[0260] (28) the sense strand comprises the sequence shown in SEQ ID NO. 783, and the antisense strand comprises the sequence shown in SEQ ID NO. 848;

[0261] (29) the sense strand comprises the sequence shown in SEQ ID NO. 783, and the antisense strand comprises the sequence shown in SEQ ID NO. 854;

[0262] (30) the sense strand comprises the sequence shown in SEQ ID NO. 784, and the antisense strand comprises the sequence shown in SEQ ID NO. 849;

[0263] (31) the sense strand comprises the sequence shown in SEQ ID NO. 831, and the antisense strand comprises the sequence shown in SEQ ID NO. 858; and

[0264] (32) The sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.869.

[0265] On the other hand, the present disclosure provides a conjugate for reducing lipoprotein a (Lp(a)) gene expression or a pharmaceutically acceptable salt thereof, comprising: (i) the aforementioned oligonucleotide or a pharmaceutically acceptable salt thereof, and (ii) a ligand conjugated to the aforementioned oligonucleotide or a pharmaceutically acceptable salt thereof, wherein at least one nucleotide of the oligonucleotide is conjugated to a targeting ligand. In some embodiments, at least one nucleotide of the aforementioned oligonucleotide or a salt thereof is conjugated to a targeting ligand to form an siRNA conjugate. The aforementioned siRNA conjugate contains the aforementioned siRNA and a conjugated group conjugated to the siRNA. The term "oligonucleotide salt" refers to an oligonucleotide compound in salt form. Oligonucleotide salts include salts of oligonucleotide conjugated compounds and salts of unconjugated oligonucleotide compounds. The oligonucleotide salt is advantageously present in the form of a solid powder.

[0266] In general, the conjugated group comprises at least one pharmaceutically acceptable targeting ligand and an optional linker, and the siRNA, linker and targeting ligand are connected in sequence. The targeting group can be a ligand conventionally used in the field of siRNA administration, such as the various ligands described in WO2009082607A2, the entire disclosure of which is incorporated herein by reference. In some embodiments, the targeting ligand is 2-4. The siRNA molecule can be non-covalently or covalently conjugated to the conjugated group, for example, it can be covalently conjugated to the conjugated group. The conjugation site of the siRNA and the conjugated group can be at the 3' end or 5' end of the siRNA sense strand or antisense strand, or can be in the internal sequence of the siRNA. In some embodiments, the conjugation site of the siRNA and the conjugated group is at the 3' end or 5' end of the siRNA sense strand. In some embodiments, the conjugation site of the siRNA and the conjugated group is at the 3' end or 5' end of the siRNA antisense strand. In some preferred embodiments, the conjugation site of the siRNA and the conjugated group is at the 3' end of the siRNA sense strand.

[0267] In some embodiments, the targeting ligand comprises an asialoglycoprotein receptor ligand. In some embodiments, the asialoglycoprotein receptor ligand comprises or consists of one or more galactose derivatives. As used herein, the term "galactose derivative" includes galactose and lactose 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-propionyl-galactosamine, N-n-butyryl-galactosamine, and N-isobutyrylgalactosamine. Galactose derivatives and clusters of galactose derivatives 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(s) facilitates cell-specific targeting to hepatocytes and endocytosis of molecules into hepatocytes. ASGPR ligands can be monomeric (eg, having a single galactose derivative) or polymeric (eg, having multiple galactose derivatives). Galactose derivatives or clusters of galactose derivatives can be linked to the 3' or 5' end of the siRNA using methods known in the art.

[0268] In some embodiments, the pharmaceutically acceptable targeting ligand in the siRNA conjugate can be galactose or N-acetylgalactosamine (GalNAc), wherein the galactose or N-acetylgalactosamine molecule can be monovalent, divalent, trivalent, or tetravalent. It should be understood that the monovalent, divalent, trivalent, and tetravalent refer to the formation of an siRNA conjugate by a conjugated group containing a galactose or N-acetylgalactosamine molecule as a targeting ligand, and the molar ratio of the siRNA molecule to the galactose or N-acetylgalactosamine molecule in the siRNA conjugate is 1: 1, 1: 2, 1: 3, or 1: 4. In some embodiments, the pharmaceutically acceptable targeting ligand is N-acetylgalactosamine. In some embodiments, when the siRNA described in the present disclosure is conjugated to a conjugated group containing N-acetylgalactosamine, the N-acetylgalactosamine molecule is trivalent or tetravalent. In some embodiments, when the siRNA described herein is conjugated to a conjugation group containing N-acetylgalactosamine, the N-acetylgalactosamine molecule is trivalent.

[0269] In some embodiments, at least one nucleotide of the oligonucleotide is conjugated to one or more targeting ligands.

[0270] In some embodiments, the targeting ligand comprises a carbohydrate, an amino sugar, cholesterol, a polypeptide, or a lipid.

[0271] In some embodiments, the targeting ligand comprises an N-acetylgalactosamine (GalNAc) moiety.

[0272] In some embodiments, the GalNac moiety is a monovalent GalNAc moiety, a divalent GalNAc moiety, a trivalent GalNAc moiety, or a tetravalent GalNAc moiety.

[0273] In some embodiments, the targeting ligand is selected from L96, A1, A3 and NAG25s. In some embodiments, the targeting ligand is located at the 5' and / or 3' end of the sense strand and / or antisense strand. In some preferred embodiments, the targeting ligand is located at the 3' end of the sense strand.

[0274] The present disclosure also provides a conjugate for reducing lipoprotein a (Lp(a)) gene expression, or a pharmaceutically acceptable salt thereof, comprising: (i) an oligonucleotide or a pharmaceutically acceptable salt thereof, and (ii) one or more lipophilic moieties conjugated to one or more internal positions on at least one chain and / or one or more lipophilic moieties conjugated to the 3' and / or 5' termini on at least one chain, optionally via a linker or a carrier.

[0275] In certain embodiments, the lipophilic portion is aliphatic, cyclic, e.g., alicyclic, or polycyclic, e.g., polyalicyclic compounds, such as steroids (e.g., sterols) or straight or branched aliphatic hydrocarbons. The lipophilic portion can typically comprise a hydrocarbon chain, which can be cyclic or acyclic. The hydrocarbon chain can comprise various substituents and / or one or more heteroatoms, such as oxygen or nitrogen atoms. Such lipophilic aliphatic portions include, but are not limited to, saturated or unsaturated C4-C 30 Hydrocarbons (e.g. C6-C 22 hydrocarbons), saturated or unsaturated fatty acids, waxes (e.g., monoalcohol esters of fatty acids and fatty diamides), terpenes (e.g., C 10 Terpenes, C 15 Sesquiterpenes, C 20 Diterpenes, C 30 Triterpenes and C 40 Tetraterpenes) and other polyaliphatic hydrocarbons. For example, the lipophilic portion may contain C4 to C 30 Hydrocarbon chains (e.g., C4 to C 30 In some embodiments, the lipophilic moiety comprises a saturated or unsaturated C6 to C 18 Hydrocarbon chain (e.g., straight chain C6 to C 22 In one embodiment, the lipophilic portion comprises a saturated or unsaturated C 16 Hydrocarbon chains (e.g., straight chain C 16 The lipid portion is a 2'-O-alkyl group, consisting of 10-30 hydrocarbon chains, including single and branched chains.

[0276] The lipophilic moiety is conjugated to the double-stranded RNAi agent through a linker comprising an ether, thioether, urea, carbonate, amine, amide, maleimide-thioether, disulfide, phosphodiester, sulfonamide bond, click reaction (e.g., triazole from azide-alkyne cycloaddition), or carbamate.

[0277] In another aspect, the present disclosure provides a composition comprising an oligonucleotide or a pharmaceutically acceptable salt thereof, or a conjugate thereof, or a salt of its conjugate, and optionally a pharmaceutically acceptable carrier.

[0278] In some embodiments, the composition is in the form of oral administration, intravenous injection, subcutaneous injection or intramuscular injection, preferably subcutaneous injection.

[0279] In some embodiments, the composition further comprises other drugs for treating and / or preventing Lp(a)-related diseases.

[0280] In some embodiments, the Lp(a)-related disease is selected from Berger'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, hyperapolipoprotein betalipoproteinemia, cerebrovascular atherosclerosis, cerebrovascular disease, venous thrombosis, stroke, atherosclerosis, thrombosis, coronary heart disease or aortic stenosis and / or any other disease or pathology associated with elevated levels of Lp(a)-containing particles.

[0281] In another aspect, the present disclosure provides use of an oligonucleotide or a pharmaceutically acceptable salt or composition thereof in the preparation of a medicament for treating and / or preventing an Lp(a)-related disease.

[0282] In some embodiments, administration of an oligonucleotide as described herein results in a decrease in the level of Lp(a) expression in a cell. In some embodiments, the decrease in the level of Lp(a) expression can be a decrease to 1% or less, 5% or less, 10% or less, 15% or less, 20% or less, 25% or less, 30% or less, 35% or less, 40% or less, 45% or less, 50% or less, 55% or less, 60% or less, 70% or less, 80% or less, or 90% or less compared to an appropriate control level of Lp(a). An appropriate control level can be the level of Lp(a) expression in a cell or cell population that has not been contacted with an oligonucleotide as described herein. In some embodiments, the effect of delivering an oligonucleotide to a cell according to the methods disclosed herein is evaluated after a limited period of time. For example, the level of Lp(a) in the blood can be analyzed at least 8 hours, 12 hours, 18 hours, 24 hours; or at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks after the oligonucleotide is introduced into the cells.

[0283] In some embodiments, inhibition of in vivo expression is determined by knocking down a human gene in a rodent expressing the human gene, e.g., an AAV-infected mouse expressing a human target gene (i.e., Lp(a)), e.g., when administered as a single dose, e.g., after subcutaneous injection at 2 mg / kg, the lowest level of Lp(a) expression is used to confirm the inhibitory effect on the human gene. Such a system is useful when the nucleic acid sequences of the human gene and the model animal gene are sufficiently close so that the human iRNA provides effective knockdown of the model animal gene.

[0284] Inhibition of Lp(a) gene expression can be indicated by a decrease in the amount of mRNA expressed by a cell line (such cells can be present, for example, in a sample derived from a subject) in which the Lp(a) gene is transcribed and which is treated (e.g., by contacting one or more cells with an iRNA disclosed herein, or by administering an iRNA disclosed herein to a subject in which cells are or were present) such that expression of the Lp(a) gene is inhibited compared to an untreated cell line that is substantially identical to the cell line (control cells that have not been treated with the iRNA or have not been treated with an iRNA targeting the gene of interest). In a preferred embodiment, inhibition is assessed using a 10 nM siRNA concentration in a species-matched cell line as described in Example 2, and is expressed as 2^-ΔΔCT of the mRNA expression level in the treated cells relative to the mRNA level in the control cells, using the following formula, wherein a housekeeping gene (e.g., GAPDH) is used as an internal reference for normalization:

[0285] △CT=CT Lp(a) -CT GAPDH

[0286] △△CT=△CT 处理细胞 -△CT 对照细胞

[0287] mRNA level = 2^-△△CT

[0288] In other embodiments, inhibition of Lp(a) gene expression can be assessed based on a decrease in a parameter functionally associated with Lp(a) gene expression, e.g., Lp(a) protein levels in blood or serum from a subject. Lp(a) gene silencing can be determined in any cell expressing Lp(a), whether endogenous or heterologous from an expression construct, and by any assay known in the art.

[0289] Inhibition of Lp(a) protein expression can be demonstrated by a decrease in the level of Lp(a) protein expressed by a cell or cell population or in a sample from a subject (e.g., the level of protein in a blood sample from a subject). As described above, to assess mRNA inhibition, inhibition of protein expression levels in treated cells or cell populations can be similarly expressed as a percentage of the protein level in a control cell or cell population, or as a change in protein level in a sample from a subject (e.g., blood or serum from a subject).

[0290] mRNA inhibition percentage = (protein expression 处理细胞 -Protein expression 对照细胞 ) / protein expression 对照细胞 *100%

[0291] Control cells, cell populations, or subject samples that can be used to assess inhibition of Lp(a) gene expression include cells, cell populations, or subject samples that have not been contacted with the RNAi agents of the present disclosure. For example, control cells, cell lines, or subject samples can be derived from individual subjects (e.g., human or animal subjects) prior to treating the subject or an appropriately matched group control with an RNAi agent.

[0292] In some embodiments of the disclosed methods, the iRNA is administered to a subject so that the iRNA is delivered to a specific site within the subject. Inhibition of Lp(a) expression can be assessed by measuring the level or change in Lp(a) mRNA or Lp(a) protein or fusion secreted luciferase in a sample of fluid or tissue from a specific site (e.g., liver or blood) of the subject.

[0293] In certain aspects, the present disclosure provides methods for preventing a disease, disorder, symptom, or condition as described herein in a subject by administering a therapeutic agent (e.g., an oligonucleotide or a vector encoding the same or a transgene) to the subject. In some embodiments, the subject to be treated is one who would benefit therapeutically from, for example, a reduction in the amount of Lp(a) protein.

[0294] The methods described herein generally involve administering to a subject an effective amount (i.e., an amount capable of producing the desired therapeutic result) of an oligonucleotide. A therapeutically acceptable amount can be an amount capable of treating a disease or condition. The appropriate dosage for any one subject will depend on certain factors, including the subject's size, body surface area, age, the specific composition to be administered, one or more active ingredients in the composition, the time and route of administration, overall health, and other drugs administered concurrently.

[0295] In some embodiments, any of the compositions disclosed herein is administered to a subject enterally (e.g., orally, through a gastric feeding tube, through a duodenal feeding tube, via gastrostomy, or rectally), parenterally (e.g., subcutaneously, intravenously, intraarterially, intramuscularly), topically (e.g., epidermally, by inhalation, via eye drops, or through a mucous membrane), or by direct injection into a target organ (e.g., the subject's liver). Typically, the oligonucleotides disclosed herein are administered intravenously or subcutaneously.

[0296] In some embodiments, the oligonucleotide is administered at a dosage in the range of about 0.001 mg / kg to about 200 mg / kg (e.g., about 0.1 mg / kg to about 100 mg / kg). In some embodiments, the oligonucleotide is administered to a subject at a dosage in the range of about 0.1 mg / kg to about 50 mg / kg, preferably about 0.1 mg / kg to about 20 mg / kg, 0.3 mg / kg to about 18 mg / kg, 0.5 mg / kg to about 15 mg / kg, or 0.5 mg / kg to about 12 mg / kg, more preferably about 1 mg / kg to about 10 mg / kg.

[0297] In some embodiments, the oligonucleotide is administered at a fixed dose of about 50 mg to about 800 mg. In some embodiments, the oligonucleotide is administered to a subject at a fixed dose of about 50 mg to about 200 mg, about 200 mg to about 400 mg, or about 400 mg to about 800 mg. In some embodiments, the oligonucleotide is administered to a subject at a fixed dose of about 100 mg, about 200 mg, about 300 mg, about 400 mg, 500 mg, about 600 mg, about 700 mg, or about 800 mg.

[0298] As a set of non-limiting examples, the oligonucleotides of the disclosure will typically be administered yearly, twice yearly, quarterly (once every three months), bimonthly (once every two months), monthly, or weekly.

[0299] In some embodiments, the fixed dose is administered to the subject at monthly intervals. In some embodiments, the fixed dose is administered to the subject at six-month intervals.

[0300] In some embodiments, a subject is administered a fixed dose of about 150 mg approximately once every six months. In some embodiments, a subject is administered a fixed dose of about 200 mg approximately once every six months. In some embodiments, a subject is administered a fixed dose of about 300 mg approximately once every six months. In some embodiments, a subject is administered a fixed dose of about 600 mg approximately once every six months. In some embodiments, a subject is administered a fixed dose of about 800 mg approximately once every six months. In some embodiments, a subject is administered a fixed dose of about 800 mg approximately once every six months.

[0301] In some embodiments, the subject to be treated is a human (e.g., a human patient) or a non-human primate or other mammalian subject. Other exemplary subjects include domestic animals such as dogs and cats; livestock such as horses, cows, pigs, sheep, goats, and chickens; and animals such as mice, rats, guinea pigs, and hamsters.

[0302] In another aspect, the present disclosure provides a method for inhibiting Lp(a) gene expression in a subject, comprising administering to the subject a fixed dose of about 50 mg to about 800 mg of an oligonucleotide or a salt thereof.

[0303] In another aspect, the present disclosure provides a method for treating a subject having an Lp(a)-related disorder comprising administering to the subject a fixed dose of about 50 mg to about 800 mg of an oligonucleotide or a salt thereof.

[0304] In another aspect, the present disclosure provides a method for lowering blood pressure levels in a subject, comprising administering to the subject a fixed dose of about 50 mg to about 800 mg of an oligonucleotide or a salt thereof.

[0305] In some embodiments, the aforementioned methods further comprise administering to the subject an additional therapeutic agent for treating an Lp(a)-related disease.

[0306] For purposes of clarity and conciseness, features are described herein as part of the same or separate embodiments; however, it will be understood that the scope of the present disclosure may include embodiments having a combination of all or some of the described features.

[0307] Hereinafter, the present disclosure will be described in more detail with reference to specific examples. However, the examples are for illustrative purposes only and have no limiting effect on the present disclosure.

[0308] Example

[0309] Abbreviation for nucleotide monomers used in nucleic acid sequence representation.

[0310] Table A. Nucleotide monomer abbreviations used in nucleic acid sequence representation

[0311] Preparation of targeting ligands

[0312] (i) L96 was prepared according to the method described in patent CN104717982B.

[0313] (ii) The preparation steps of A1 are as follows:

[0314] (1) Synthesis of intermediate E1

[0315] F1 (199.46 mg, 0.41 mmol) was dissolved in DMF and activated with HBTU (155.39 mg, 0.41 mmol). DIEA (67.81 L, 0.41 mmol) was then added and stirred for 30 min. Intermediate G1 (100.0 mg, 0.14 mmol) was added to the reaction system. After stirring overnight, the reaction was complete by TLC. After concentration under reduced pressure, column chromatography (PE:EA = 10:1) was performed to obtain intermediate I-F1 (174.98 mg, 58% yield). Intermediate I-F1 was then treated with palladium on carbon under hydrogen catalysis to remove the benzyl group to obtain intermediate II-F1 (156.34, 95%). HBTU-activated intermediate II-F1 was coupled with (3R,5S)-5-[[bis(4-methoxyphenyl)phenylmethoxy]methyl]-3-pyrrolidinol in DMF to obtain intermediate E1. 1 HNMR (400MHz, DMSO-D6): δ7.91-7.86(m,8H),7.51-7.45(m,6H),7.40-7.32(m,10H),7.02-6.95(m,3H),6.03-5.89( m,4H),5.31(d,3H),5.07-4.99(m,4H),4.43-4.37(m,3H),4.23-4.19(m,4H),4.17-4.13(m,6H),4.05-4.00(m,6H),3 .88-3.81(m,6H),3.77(s,6H),3.69-3.66(m,9H),3.50-3.47(m,12H),3.40-3.37(m,6H),3.20-3.16(m,16H),2.60- 2.57(m,2H),2.46(s,9H),2.30(s,9H),2.10(m,6H),2.07(s,9H),1.98(s,9H),1.56-1.45(m,8H),1.32-1.27(m,4H). HRMS[M+H] + m / z 2467 (calcd for C 115 H 168 N 14 O 45 ,2466).

[0316] (2) Synthesis of solid phase support A1 with protective groups

[0317] Intermediate E1 was treated with succinic anhydride in the presence of DMAP and Et3N to obtain the hemisuccinate intermediate E1-1 in quantitative yield. The hemisuccinate intermediate E1-1 was coupled with the amino groups of a 60% divinylbenzene (DVB) cross-linked (aminomethyl) polystyrene resin with an amine content of 250 mol / g to obtain a solid support A1 with a loading of 100 mol / g and a protecting group.

[0318] (iii) The preparation steps of A3 are as follows:

[0319] Intermediate E3 was prepared by a similar method to that of E1. 1 HNMR(400MHz,DMSO-D6):8.79-8.65(m,6H),7.85-7.75(m,9H),7.44-7.35(m,12H),7.02-6.94(m,3H),5.99-5.89(m ,4H),5.15(d,3H),5.01-4.95(m,4H),4.46-4.33(m,4H),4.25-4.17(m,8H),4.13-4.07(m,10H),4.05-3.97(m,10H), 3.90-3.80(m,9H),3.77(s,6H),3.73-3.66(m,8H),3.56-3.50(m,14H),3.41-3.36(m,6H),3.22-3.17(m,14H),2.59- 2.56(m,2H),2.47(s,9H),2.32(s,9H),2.14(m,6H),2.08(s,9H),2.00(s,9H),1.57-1.45(m,8H),1.33-1.26(m,4H). HRMS[M+H] + m / z 2810 (calcd for C 127 H 186 N 20 O 51 ,2809)

[0320] A3 was prepared via E3 in a similar manner to that of A1.

[0321] Preparation of oligonucleotides

[0322] (1) Preparation of siRNA

[0323] The siRNA sequence is synthesized separately on a solid support via a sense strand (SS) and an antisense strand (AS), and is obtained after deprotection, cleavage, purification, annealing, purification, and lyophilization.

[0324] [Corrected 02 / 12 / 2024 according to Rule 91] Solid-phase synthesis: Sense and antisense strands are synthesized separately on a solid support using phosphoramidite technology using an automated oligonucleotide synthesizer. Examples of such synthesizers include the AKTA Oligopilot (Cytiva) and the Dr. Oligo 192XLc (Kunshan Berlik Precision Instrument Co., Ltd.). Solid-phase synthesis begins at the 3' end of the sequence and couples monomers sequentially into the sequence. Each coupling of a phosphoramidite monomer involves four chemical steps: 1) unblocking or deprotection (removal of the hydroxyl protecting group); 2) coupling; 3) oxidation; and 4) capping. All phosphoramidite monomers, reagents, and purification consumables used were commercially available, including various phosphoramidite monomers (e.g., 5'-O-(4,4'-Dimethoxytrityl)-2'-O-methyl-Uridine-3'-CE-hosphoramidite) purchased from Shanghai Zhaowei Technology Development Co., Ltd., and reaction reagents (e.g., 40 wt% aqueous methylamine solution, 28 wt% aqueous ammonium hydroxide solution) purchased from Sigma-Aldrich LLC. The siRNA synthesis and purification methods used herein are described in US20130178612A1 and US2015100197A1, among others; the synthesis methods for sequences containing VPUm and APU structures are described in J. Med. Chem. 2018, 61, 734-744.

[0325] (2) Preparation of double-stranded RNA reagent

[0326] (a) Synthesis of the positive chain

[0327] The solid-phase phosphoramidite method is a well-established method for oligonucleotide synthesis. It utilizes a computer-controlled synthesizer and is carried out within a stainless steel synthesis column. Sense strand synthesis begins with a solid support loaded with a targeting ligand (e.g., L96 and A1), or directly from the solid support. The solid-phase synthesizer controls different pipelines, injecting various raw materials, reagents, and solvents in a 3' to 5' sequence order, ligating phosphoramidite nucleoside monomers one by one. The reaction involves four cycles: DMT protection group removal, condensation, oxidation or thiolation, and capping. Each cycle adds a single nucleotide unit to the oligonucleotide, yielding sequences of 19 or 21 nucleotide units. Following synthesis, the protecting group (2-cyanoethyl) is removed on the solid-phase synthesis column, and the synthesized sequence is cleaved from the solid support via aminolysis. The resulting product is filtered, the filter cake washed with ethanol, and the filtrate and washings are collected and concentrated to yield the crude sense strand. The crude product is purified by chromatography (SOURCE 15Q) and lyophilized to yield the desired sense strand. Among them, in the synthesizer, the siRNA sense chain conjugate is synthesized starting from the solid support loaded with the targeting ligand (such as L96); and the siRNA is synthesized directly starting from the solid support.

[0328] (b) Synthesis of antisense strand

[0329] The synthesis of the antisense strand is similar to that of the sense strand. A solid-phase synthesizer controls different pipelines to inject different raw materials, reagents, and solvents in the order of 3' to 5' of the sequence, connecting the phosphoramidite nucleoside monomers one by one. The reaction process includes four cycles: DMT protection group removal, condensation reaction, oxidation or thiolation reaction, and end-capping reaction. Each cycle connects a nucleotide unit to obtain an oligonucleotide sequence of 21 or 23 nucleotide units. After synthesis, the protecting group (2-cyanoethyl) is removed on a solid-phase synthesis column, and the synthesized sequence is cleaved from the solid phase support by aminolysis. The residue is filtered, the filter cake is washed with ethanol, and the filtrate and washing liquid are collected and concentrated to obtain the crude antisense strand. The crude product is purified by chromatography (SOURCE 15Q), ultrafiltration, and lyophilization to obtain the target antisense strand siRNA.

[0330] (c) Preparation of double-stranded siRNA

[0331] Dissolve the AS and SS strands separately in injection water, mix at a defined ratio (1.01:1.0-1.2:1.0), incubate at 30-50°C for 30-90 minutes, cool to room temperature, and freeze-dry to obtain double-stranded siRNA.

[0332] According to the same method, the double-stranded siRNA reagents shown in Tables 2, 3 and 4 below were prepared.

[0333] Example 1. Activity screening of naked Lp(a)-siRNA sequences in primary cynomolgus monkey liver cells

[0334] First, a computer-based algorithm was used to generate candidate oligonucleotide sequences complementary to human Lp(a) mRNA (NM_005577.4, Table 1). Some of these sequences were also complementary to or had no more than two mismatches with cynomolgus macaque Lp(a) mRNA (XM_045391712.1, Table 1). Some of these sequences were designed as double-stranded siRNAs with 19 / 21 pairings between the sense and antisense strands, with the antisense strand having two overhangs complementary to the mRNA sequence, or in some cases, non-complementary UU overhangs. Some of these sequences were designed as double-stranded siRNAs with 21 / 23 pairings between the sense and antisense strands, with the antisense strand having two overhangs complementary to the mRNA sequence. Some of these sequences were designed as double-stranded siRNAs with 21 / 21 or 23 / 23 pairings. In some complementary paired sequences, the first base at the 5' end of the antisense strand (the last base at the 3' end of the sense strand) is replaced by a base that does not match Lp(a) mRNA.

[0335] Table 1: Human and cynomolgus monkey Lp(a) mRNA sequences

[0336] In Tables 2, 3, and 4, "G," "C," "A," "U," and "T" generally represent nucleotides based on guanine, cytosine, adenine, uracil, and thymine, respectively. dA represents 2'-deoxyadenosine 3'-phosphate; dG represents 2'-deoxyguanosine 3'-phosphate; dC represents 2'-deoxycytidine 3'-phosphate; and dT represents 2'-deoxythymidine 3'-phosphate.

[0337] For nucleotide modifications: m represents 2'-methoxy; f represents 2'-deoxy-2'-fluoro; and s represents phosphorothioate.

[0338] Certain oligonucleotides may include a diol nucleic acid (GNA) modification, wherein the ribonucleotide has a diol nucleic acid structure as shown in formula (I), wherein Base represents a nucleic acid base selected from A, G, C, and T. In certain oligonucleotides, individual ribonucleotides are replaced with (Tgn), which is a thymidine-diol nucleic acid (GNA) S-isomer as shown in formula (II). In certain oligonucleotides, individual ribonucleotides are replaced with (Cgn), which is a cytidine-diol nucleic acid (GNA) S-isomer as shown in formula (III). In certain oligonucleotides, individual ribonucleotides are replaced with (Agn), which is an adenosine-diol nucleic acid (GNA) S-isomer as shown in formula (IV). In certain oligonucleotides, individual ribonucleotides are replaced with (Ggn), which is a guanosine-diol nucleic acid (GNA) S-isomer as shown in formula (V). The structural formulas of GNA, Tgn, Cgn, Agn, and Ggn are as follows:

[0339] Certain oligonucleotides may comprise 2'-5'-phosphodiester bonds, wherein adjacent pairs of nucleoside units are linked 2'-5' to 5'-2'. Ribonucleotides have a nucleoside-2'-phosphate structure as shown in formula (VI), wherein Base represents a nucleic acid base selected from A, G, C, and U. In certain oligonucleotides, individual ribonucleotides are replaced by (U-2'5'), which is a uridine-2'-phosphate as shown in formula (VII). In certain oligonucleotides, individual ribonucleotides are replaced by (C-2'5'), which is a cytidine-2'-phosphate as shown in formula (VIII). In certain oligonucleotides, individual ribonucleotides are replaced by (A-2'5'), which is an adenosine-2'-phosphate as shown in formula (IX). In certain oligonucleotides, individual ribonucleotides are replaced by (G-2'5'), which is a guanosine-2'-phosphate as shown in formula (X). The structural formulas of nucleoside-2'-phosphates, U-2'5', C-2'5', A-2'5' and G-2'5' are as follows:

[0340] Some oligonucleotides have a 5'-phosphate analog modified nucleotide at the 5' end. In some oligonucleotides, the 5'-phosphate analog modified nucleotide has a vinyl phosphonate modified nucleotide as shown in formula (XI). In some oligonucleotides, the 5'-phosphate analog modified nucleotide has a vinyl phosphonate modified nucleotide as shown in formula (XII). Wherein, R is selected from H, OH, fluorine, 2'-methoxy, 2'-acetylamino, 2'-aminoethyl and 2'-O-methoxyethyl, and Base represents a nucleic acid base selected from A, G, C and U. In some oligonucleotides, the 5' end has APU, which is a 5'-phosphate analog modified uridylic acid (2'-acetylamino-5'-vinylphosphonate-uridylic acid) as shown in formula (XIII). In some oligonucleotides, the 5' end has VPU, which is a 5'-phosphate analog modified uridylic acid (2'-methoxy-5'-vinylphosphonate-uridylic acid) as shown in formula (XIV).

[0341] Certain oligonucleotides may comprise inverted ribonucleotides (3'-3' linked nucleotides) of formula (XV). In certain oligonucleotides, individual ribonucleotides are replaced with (invdA), which is an inverted deoxyadenylate of formula (XVI).

[0342] In certain oligonucleotides, individual ribonucleotides are replaced with M, which is 2'-O-methyl-6-(3-(2-carboxyethyl)phenyl)-adenylic acid as shown in formula (XVII).

[0343] Certain oligonucleotides can be conjugated to targeting ligands. Targeting ligands are selected from L96, A1, A3, and NAG25s, wherein L96 is N-[tris(GalNAc-alkyl)amidodecanoyl]-4-hydroxyprolinol (Hyp-(GalNAc-alkyl)3); A1, A3, and NAG25s are GalNAc carriers. The structural formulas of L96, A1, A3, and NAG25s are as follows:

[0344] Table 2: Naked oligonucleotide sequences

[0345] Table 3: Modified oligonucleotides

[0346] Table 4: siRNA sequences with vectors

[0347] (1) Cynomolgus monkey hepatocyte cell culture and transfection:

[0348] Before adding cells, remove culture medium (Beijing Red Biotech Co., Ltd., HEPO24) and preheat in a biosafety cabinet. Add 4 ml of FBS to 36 ml of culture medium to prepare complete medium and warm in a 37°C water bath for 10 min. Treat with coating medium (Beijing Red Biotech Co., Ltd., HEPO44) in a CO2 incubator at 37°C for 0.5 h. Remove primary cynomolgus macaque hepatocytes (Beijing Red Biotech Co., Ltd., cmTCSC) from liquid nitrogen and thaw in a 37°C water bath. After approximately 2 min, remove the cells and transfer the cell suspension to 40 ml of preheated complete medium. Rinse the cryovial with 2 ml of complete medium and centrifuge the cell suspension at 180 × g for 1 min. Discard the supernatant and add 2 ml of preheated CM seeding medium (Beijing Red Biotech Co., Ltd., CMHEP054). Gently pipette the cell suspension to mix thoroughly. Count 20 μl of the cell suspension. Based on the count results, cells were diluted to 3*10^5 / ml and seeded into 12-well plates at 3*10^5 / well. The plates were then cultured in a 37°C, 5% CO2 incubator. After 4-5 hours of adherence, the CM seeding medium was aspirated and replaced with pre-warmed culture medium (Beijing Red Biotech Co., Ltd., CMHEP064) at 0.9ml / well. Transfection was performed 6 hours after adherence. Lipofectamine was used. TM Transfection was performed using 3000 Transfection Reagent (thermofisher, L3000150). System ① used 50 μl Opti-MEM (thermofisher, 1105821) to dilute the siRNA (Suzhou Ouli Biopharmaceutical Technology Co., Ltd.) to a concentration of 1 μM, 0.2 μM, or 20 nM. System ② used 50 μl Opti-MEM to dilute 3 μl Lipofectamine TM 3000Transfection Reagent, after standing for 5 minutes, mix system ① and ②, the final concentration of siRNA is 0.5μM or 0.1μM, stand for 15 minutes, add dropwise to the 12-well plate, the final concentration of siRNA is 50nM, 10nM or 1nM, replace DMEM / F12 complete medium 4 hours after transfection, and place the 12-well plate in an incubator for incubation for 24 hours.

[0349] (2) RNA extraction and detection

[0350] (i) Total RNA was extracted using RNA-Quick Purification Kit (RN001, Yishan Biotechnology):

[0351] Remove the 12-well plate from the incubator, aspirate the medium, wash once with an appropriate amount of PBS, add 500 μl of lysis buffer to each well, and transfer the supernatant to a fresh 1.5 ml centrifuge tube. Add 500 μl of absolute ethanol to the lysed cells and mix thoroughly (if a precipitate forms, this is normal; continue with the procedure). Invert the centrifuge tube several times or pipette vigorously 10 times to disperse the precipitate. Then add the liquid to the spin column, place the tubes symmetrically in a centrifuge (Eppendorf, 5430), and centrifuge at 4000 × g for 1 minute. Remove the centrifuge tube and add 500 μl of wash buffer to the column. Centrifuge at 12000 × g for 1 minute. After centrifugation, remove the column, discard the waste liquid, and return the RNA column to the collection tube. Centrifuge the empty tube once to remove any residual wash buffer. Place the column in a clean RNase-free 1.5 ml centrifuge tube and air dry it with the lid open for 2 minutes. Add 30 μl of elution buffer to the center of the RNA column membrane. Let stand at room temperature for 2 minutes. Centrifuge at 2000 × g for 1 minute to elute the RNA, then place on ice. Measure the eluted RNA concentration for subsequent experiments. The extracted RNA can be used immediately for subsequent experiments or stored at -80°C for later use.

[0352] (ii) Use cDNA was synthesized using IIQ RT SuperMix for qPCR (+gDNA wiper) reverse transcription kit (Novozyme, R223-01):

[0353] In an RNase-free centrifuge tube, prepare a mixture of 4 μl of 4×g DNA wiper mix, 1 μg of template RNA, and 16 μl of RNase-free ddH₂O to remove genomic DNA. Gently pipette to mix thoroughly and incubate at 42°C for 2 minutes. Then, add 4 μl of 5× HiScript II qRT SuperMix II directly to the reaction tube and gently pipette to mix thoroughly. Place the tube in a PCR instrument (Applied Biosystems, 9700) and cycle at 50°C for 15 minutes, 85°C for 5 seconds, and hold at 4°C. The product can be used immediately for qPCR reactions or stored at -20°C and used within six months. For long-term storage, aliquot and store at -80°C. Avoid repeated freeze-thaw cycles for cDNA.

[0354] (iii) qPCR quantification using ChamQ SYBR qPCR Master Mix (Novozymes, Q311-02):

[0355] A mixture of 10 μl 2× ChamQ SYBR qPCR Master Mix, 0.5 μl Forword primer (Ruibo Kexing), 0.5 μl Reverse primer (Ruibo Kexing), 1 μl Template cDNA, and 8 μl ddH2O was prepared to 20 μl system. Each sample was replicated three times. The 96-well plate was placed in a qPCR instrument (ROCGENE, Archimed) and the program was executed: pre-deformation, 95°C, 30 sec; amplification, 95°C, 10 sec, 60°C, 30 sec, 40 cycles; melting curve, 95°C, 15 sec, 60°C, 60 sec, 95°C, 15 sec.

[0356] (3) Data statistical analysis:

[0357] Export the data to EXCEL format and use CT LPA -CT GAPDH The control group was normalized. In order to calculate the fold change of relative silencing efficiency, the data were analyzed using the △△CT method, and the mean and standard deviation of the three parallel repeated data were calculated.

[0358] The results of the three screenings of monkey primary cells are shown in Tables 5 to 8.

[0359] As shown in Table 5, when the dosage was 50 nM, AL0111001, AL0111004, AL0111005, AL0111007, AL0111008, AL0111009, AL0111010, AL0111011, AL0111013, AL0111014, AL0111016, AL0111017, AL0111018, AL0111019, AL0111020 The inhibitory effect of siRNAs for AL0111021, AL0111022, AL0111026, AL0111027, AL0111029, AL0111030, AL0111031, AL0111032, AL0111033, AL0111034, AL0111035, AL0111036, AL0111037, AL0111038, and AL0111044 was ≥90%.

[0360] Table 5: Lp(a) siRNA naked sequence knockdown levels in monkey primary hepatocytes

[0361] After ranking mRNA inhibition rates from high to low, siRNA sequences with higher inhibition rates were designed based on sequence position. The knockdown of Lp(a) mRNA by naked sequences AL0111053-AL0111068 at a dose of 10 nM is shown in Table 6. Using AL0111044 as a control, all sequences achieved knockdown rates greater than 90%, even exceeding those of the control sequence.

[0362] Table 6: Lp(a) siRNA naked sequence knockdown levels in monkey primary hepatocytes

[0363] After ranking the knockdown levels of siRNAs AL0111001-AL0111068, the optimal sequences were selected for activity testing at a low concentration of 1 nM. The results are shown in Table 7. As shown in Table 7, with the exception of AL0111056, AL0111058, and AL0111059, the knockdown rates of Lp(a) mRNA at a concentration of 1 nM were ≥50%. Among them, AL0111009, AL0111053, AL0111054, and AL0111061 achieved knockdown rates ≥80%, indicating significant efficacy.

[0364] Table 7: Lp(a)-siRNA naked sequence knockdown levels in monkey primary hepatocytes

[0365] Example 2. Screening of Lp(a)-siRNA activity in RT4 cells

[0366] (1) RT4 cell culture and transfection:

[0367] Human bladder transitional cell papilloma RT4 cells (Sybest Biotech Co., Ltd., Shanghai) were used and placed in a 37°C, 5% CO2 incubator. They were cultured in DMEM medium (Sybest Biotech Co., Ltd., Shanghai) supplemented with 10% FBS (GIBCO, 12483020) and 1% penicillin-streptomycin (GIBCO, 15140-122). When the cell confluence reached 90%, they were digested with trypsin-EDTA (Thermo, 25200-072) and counted using a cell counter (Countstar, IC1000). 190 μl of cell suspension / well was inoculated into a 96-well plate at a cell number of 2*104 cells / well and allowed to adhere to the plate the next day for transfection.

[0368] Transfection was performed using Lipofectamine™ RNAiMAX (thermofisher, 13778150). 2.2 μl (2 μM) of the diluted compound, 19.1 μl Opti-MEM, and 0.7 μl RNAiMAX were mixed to form a transfection complex. After incubation for 5 minutes, the transfection complex was added to the cells (n=2) at a final siRNA concentration of 10 nM at 10 μl per well. The cells were cultured in a 37°C, 5% CO2 incubator for 24 hours.

[0369] (2) RNA extraction and detection

[0370] 24 hours after transfection, the culture medium was removed, and the cells were washed once with PBS. 200 μl of lysis buffer was added to each well, incubated at room temperature for 15 minutes, and then vortexed to mix thoroughly. Total RNA was extracted using a 96-well automated nucleic acid extraction instrument (Hanwei Technology, HW-96 series) and a matching blood RNA magnetic bead extraction kit (Zhiang Bio, GO-MNTR).

[0371] cDNA was reverse transcribed using HiScript IIQ RT SuperMix for qPCR (with gDNA wiper) (Vazyme, R223-01). A mixture of 4 μl of 4X gDNA wiper mix and 12 μl of RNA sample was prepared in an RNase-free centrifuge tube. The mixture was then made up to 16 μl with RNase-free ddH2O. Each sample was replicated at 42°C for 2 min. Four μl of 5X HiScript IIQ RT SuperMix II was added to the first reaction tube. Mix thoroughly and incubate at 50°C for 15 min and 85°C for 5 s.

[0372] A mixture of 10 μl 2XChamQ SYBR QPCR Master MIX (Beijing Vazyme Biotechnology Co., Ltd. (Vazyme), Q311-02) was prepared in an RNase-free centrifuge tube, 2 μl of sample, 1 μl of 10 μM primer, and RNase-free ddH2O was added to 20 μl. Each sample was replicated three times. The 96-well plate was placed in a qPCR instrument (ABI, QuantStudio5 Real-Time PCR instrument) and the program was executed: stage 1, 95°C, 30 sec; amplification, 95°C, 5 sec; 60°C, 20 sec; 40 cycles; melting curve, 95°C, 15 sec, 60°C, 60 sec, 95°C, 15 sec.

[0373] (3) Data statistical analysis:

[0374] Export the data to EXCEL format and use CTLp(a) -CT GAPDH The control group was normalized, and the data were analyzed using the △△CT method to calculate the fold change of relative silencing efficiency. The results are shown in Table 8.

[0375] When the dosage was 10 nM, the inhibitory effect of sequence AL0115036-AL0115153 on the expression level of Lp(a) mRNA in RT4 cell line was shown in Table 8. Among them, AL0115019, AL0115041, AL0115042, AL0115044, AL0115045, AL0115046, AL0115047, AL0115049, AL0115052, AL0115054, AL0115055, AL0115056, AL0115057, AL0115061, AL0115062, AL0115064, AL0115065, AL0115066, AL0115067, AL0115070 68. The knockdown rate of Lp(a) mRNA by AL0115069, AL0115070, AL0115071, AL0115072, AL0115073, AL0115074, AL0115075, AL0115095, AL0115096, AL0115119, AL0115135, AL0115137, AL0115138, AL0115139, AL0115141, AL0115142 and AL0115143 was ≥40%, or even greater than 60%.

[0376] Table 8: Knockdown of Lp(a)-siRNA modified sequences in RT4 cells

[0377] Example 3. Activity screening of chemically modified Lp(a)-siRNA in primary monkey hepatocytes

[0378] (1) Cynomolgus monkey hepatocyte cell culture and transfection:

[0379] Same as Example 1.

[0380] (2) RNA extraction and detection

[0381] Same as Example 1.

[0382] (3) Data statistical analysis:

[0383] Same as Example 1.

[0384] The results of the three screenings of monkey primary cells are shown in Tables 9 to 17. Tables 9 to 17 show the results of experiments conducted at different times using different batches of monkey primary cells.

[0385] As can be seen from Tables 9 to 14, the siRNAs that showed an inhibitory effect of more than 70% on Lp(a) mRNA at a high concentration of 1 nM, or at least 50% at a low concentration of 0.1 nM were AL0115001, AL0115002, AL0115003, AL0115004, AL0115005, AL0115006, AL0115007, AL0115008, AL0115009, AL0115010, AL0115011, AL0115012, AL0115013, AL0115014, AL0115015, AL0115016, AL0115017, AL0115018, and AL0115019. 5020, AL0115026, AL0115029, AL0115031, AL0115032, AL0115034, AL0115035, AL0115 152. AL0115153, AL0115045, AL0115052, AL0115054, AL0115057, AL0115064, AL01150 68, AL0115074, AL0115096, AL0115119, AL0115137, AL0115138, AL0115162, AL0115164, AL0115169, AL0115170, AL0115178, AL0115179, AL0115180, AL0115182, and AL0115186. In particular, AL0115137, AL0115161, AL0115162, AL0115163, AL0115164, AL0115169, AL0115170, AL0115178, AL0115180, AL0115182 and AL0115186 can inhibit Lp(a) mRNA by more than 70% at a low concentration of 0.1 nM, indicating a significant inhibitory effect.

[0386] As can be seen from Tables 15-17, AL0115151, AL0115164, AL0115185, AL0115186, AL0115189, AL0115210, AL0115220, AL0115222, AL0115223, AL0115224, AL0115225, AL0115227, AL0115228, AL0115229, AL0115230, AL0115231, AL0115232, AL0115237, AL0115238 and AL0115239 can inhibit Lp(a) mRNA by more than 70%, and the inhibitory effect is significant. In particular, AL0115164, AL0115185, AL0115186, AL0115189, AL0115210, AL0115224, AL0115225, AL0115231, AL0115237, AL0115238 and AL0115239 can still achieve at least 70% target gene inhibition effect at a dosage of 0.1 nM.

[0387] Table 9: Knockdown levels of chemically modified Lp(a) siRNA in primary cynomolgus monkey hepatocytes Note: NA stands for Not Detected

[0388] Table 10: Knockdown levels of chemically modified Lp(a) siRNA in primary cynomolgus monkey hepatocytes Note: NA means not tested

[0389] Table 11: Knockdown levels of chemically modified Lp(a) siRNA in primary cynomolgus monkey hepatocytes Note: NA means not tested

[0390] Table 12: Knockdown levels of chemically modified Lp(a) siRNA in primary cynomolgus monkey hepatocytes Note: NA means not tested

[0391] Table 13: Knockdown levels of chemically modified Lp(a) siRNA in primary cynomolgus monkey hepatocytes

[0392] Table 14: Knockdown levels of chemically modified Lp(a) siRNA in primary cynomolgus monkey hepatocytes

[0393] Table 15: Knockdown levels of chemically modified Lp(a) siRNA in primary cynomolgus monkey hepatocytes

[0394] Table 16: Knockdown levels of chemically modified Lp(a) siRNA in primary cynomolgus monkey hepatocytes

[0395] Table 17: Knockdown levels of chemically modified Lp(a) siRNA in primary cynomolgus monkey hepatocytes

[0396] Example 4. In vivo testing of Lp(a) RNAi agents in Gluc mice

[0397] Gluc is easily secreted and highly sensitive, and can directly measure Gluc expression activity in whole blood. Gluc is connected to an exogenous gene by P2A, and the expression level of Gluc directly reflects the exogenous gene mRNA level. The mRNA sequence of human Lp(a) gene (GENBANK NO.NM_005577) is obtained in the NCBI database. The human Lp(a) gene 2609-2677 base sequence is selected and connected to the Gaussia secretory luciferase (Gluc) encoding gene by P2A to obtain a fusion protein gene (SEQ ID NO.870), which is secreted and expressed in cells as a fusion protein. Recombinant plasmids were obtained by conventional molecular biology techniques such as enzyme digestion and ligation. First, the sequence containing the CAG promoter (SEQ ID NO. 871) was inserted between EcoRI and XhoI of the pFB vector (purchased from Agilent, Catalog No. 013001) to obtain pFB-AAV-CAG. The fusion protein gene was then inserted between EcoRI and BamHI of the pFB-AAV-CAG vector by enzyme digestion and ligation to obtain the pFB-AAV-CAG-Gluc-2A-LPA-DS vector (Figure 1). Transient transfection with the plasmid was performed in vivo by hydrodynamic tail vein injection, which was administered at least 29 days before the administration of the Lpa RNAi agent or control.

[0398] The experiment used SPF male C57BL / 6 mice aged 6 to 8 weeks (Beijing Biotechnology Co., Ltd.). Each mouse was injected with 1.00E+11 vg / mL of pFB-AAV-CAG-Gluc-2A-LPA-DS vector via the tail vein. Then, according to the experimental design requirements, orbital blood was collected at different time points, and serum was separated for Gluc luminescence detection (Pierce TMTwo groups of experiments were performed. One group was randomly divided into a vehicle control group (NC group) based on the Lpa-Gluc luminescence detection value, and eight groups were randomly divided into test groups (AL0117001, AL0117002, AL0117003, AL0117004, AL0117005, AL0117006, and AL0117007), each containing five mice. After grouping, mice were subcutaneously administered a single dose of the corresponding LPA RNAi agent and a vehicle control. Blood samples were collected from mice on days 8, 15, 22, 36, 43, and 50 after administration (Figure 2A). Another set of experiments was randomly divided into eight groups (five mice each) based on Lpa-Gluc luminescence values: a vehicle control group (NC group) and test groups (AL0117004, AL0117005, AL0117006, AL0117007, AL0117008, AL0117009, and AL01170010). After grouping, mice were administered a single subcutaneous dose of the corresponding LPA RNAi agent and a vehicle control. Blood was collected from the mice on days 8, 15, 22, 29, and 36 after administration (Figure 2B). Serum was isolated and analyzed for luc luminescence to evaluate the inhibitory effect of the siRNA on the exogenous gene mRNA. The results are shown in Figure 2. The most significant and longest-lasting reductions were observed in the AL0117005, AL0117006, AL0117007, and AL0117009 groups (Figure 2).

[0399] Example 5. In vivo testing of Lp(a) RNAi agents in transgenic mice

[0400] The study used SPF male C57BL / 6JSmoc-Tg(pAlb-hLPA)1Smoc mice, 8-9 weeks old (purchased from Shanghai Model Organisms). Pre-dose serum samples were obtained on day 0, and the mice were randomly divided into groups based on hLp(a) levels. C57BL / 6JSmoc-Tg(pAlb-hLPA)1Smoc mice were administered a single subcutaneous dose of 1 mg / kg of the Lp(a) RNAi agents AL0117014, AL0117015, AL0117016, AL0117017, AL0117018, AL0117019, and AL0117020. A control group (NC) was established as a vehicle group and administered the same volume of 0.9% saline. Blood samples were collected from the mice (from the eyeballs and sent for analysis within 1 hour) at 1, 2, 3, 4, 5, and 6 weeks after administration. The pre-dose control was used as the control to measure hLp(a) knockdown levels. During the experiment, no animals showed signs of death or dying. Clinical observation revealed no significant abnormalities in any of the animals. Changes in hLp(a) levels are shown in Figure 3.

[0401] As shown in Figure 3, compared to the control group (NC), all Lp(a) siRNA drug groups effectively knocked down Lp(a) protein in mice. The knockdown effect of all hLp(a) siRNAs reached its lowest point 22 days after drug intervention, then gradually recovered. By day 55, blood hLp(a) levels had returned to pre-drug levels. All drug interventions significantly reduced hLp(a) levels in the mice's blood, with the AL0117015 group achieving the most significant and longest-lasting reduction.

[0402] In another set of experiments, a single subcutaneous dose of 1 mg / kg of Lp(a) RNAi agents, AL0117021, AL0117022, AL0117023, AL0117024, AL0117025, AL0117026, AL0117027, AL0117028, AL0117029, AL0117030, and AL0117016, was administered. A control group (NC) was also established, receiving the same volume of 0.9% saline. Blood samples were collected from the eyeballs of the mice at 1, 2, 3, 4, 5, and 6 weeks after administration, and hLp(a) knockdown levels were measured using pre-dose control as the control. During the experimental period, no animals showed signs of mortality or moribundity. Clinical observation revealed no significant abnormalities in any of the animals. Changes in hLp(a) levels are shown in Figure 4.

[0403] As can be seen from Figure 4, compared with the control group (NC), all drug-treated groups except AL0117023 and AL0117025 groups showed significant knockdown of hLp(a) protein, especially AL0117024, AL0117029, and AL0117030, which showed more sustained efficacy and better target knockdown efficacy during the 5-week administration.

[0404] Example 6. In vivo testing of Lp(a) RNAi agents in cynomolgus monkeys

[0405] The efficacy of AL0117030, AL0117038, and AL0117040 was evaluated in crab-eating macaques. Male crab-eating macaques aged 3-6 years (purchased from Zhaoyan Pharmaceutical) were used in the experiment. Pre-dose serum samples were obtained on day 0 of administration, and the animals were randomly divided into groups according to Lp(a) levels. All experimental groups were given 2 mg / kg of Lp(a) RNAi agent. Blood from crab-eating macaques was collected at 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 9 weeks, 11 weeks, and 14 weeks after administration, and the Lp(a) expression level was measured, and the level of Lp(a) knockdown was detected using the pre-dose control as the control. During the experiment, no obvious abnormalities were found in all animals during clinical observation. The experimental groups are shown in Table 18; the change levels of Lp(a) are shown in Figure 5.

[0406] Table 18 Dosage Grouping

[0407] As shown in Figure 5, after a single dose, AL0117030, AL0117038, and AL0117040 can continuously silence Lp(a) expression in cynomolgus macaque serum, with the highest silencing efficiency exceeding 90%. Sustained silencing of more than 80% can be maintained for at least nearly 100 days, supporting a lower frequency of clinical use in the future.

Claims

1. An oligonucleotide or a pharmaceutically acceptable salt thereof for reducing lipoprotein a (Lp(a)) gene expression, the oligonucleotide comprising a sense strand and an antisense strand, the sense strand having a sequence having at least 80% sequence identity with a sequence shown in any one of SEQ ID NOs. 1-43, 45-66, 68-97, 99-206, 735-749 and 807-817 or a fragment thereof, or a modified sequence of the sequence or a fragment thereof, preferably a sequence having 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity; the antisense strand having a sequence ... A sequence having at least 80% sequence identity to any of the sequences shown in NO. 207-249, 251-276, 278-302, 304-404, 750-769 and 818-828 or a fragment thereof, or a modified sequence of the sequence or its fragment, preferably a sequence having 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity.

2. The oligonucleotide or a pharmaceutically acceptable salt thereof according to claim 1, wherein The oligonucleotide or a pharmaceutically acceptable salt thereof is selected from carboxylate, alkali metal salt, ammonium salt, alkaline earth metal salt, salt formed with an organic base and other pharmaceutically acceptable salts; Preferably, the salt is an alkali metal salt, more preferably a sodium salt or a potassium salt thereof; Preferably, the salt is an alkaline earth metal salt, more preferably a magnesium salt or a calcium salt; Preferably, the salt is an ammonium salt, more preferably a triethylamine salt.

3. The oligonucleotide or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein: The oligonucleotide comprises at least one modified nucleotide; Preferably, the oligonucleotide comprises at least one 2'-modified nucleotide; Preferably, the 2'-modified nucleotide is selected from one or more of 2'-alkoxy modified nucleotides, 2'-substituted alkoxy modified nucleotides, 2'-alkyl modified nucleotides, 2'-substituted alkyl modified nucleotides, 2'-amino modified nucleotides, 2'-substituted amino modified nucleotides, 2'-fluoro modified nucleotides, and 2'-deoxy nucleotides; Preferably, the 2'-modification is a modification selected from the group consisting of: 2'-methoxy, 2'-acetylamino, 2'-aminoethyl, 2'-fluoro, 2'-O-methoxyethyl; Preferably, the oligonucleotide has a 5'-phosphate analog modified nucleotide at the 5' terminal nucleotide; preferably, the 5'-phosphate analog modified nucleotide has a vinyl phosphonate modified nucleotide shown in formula (XII), wherein R is selected from H, OH, fluorine, 2'-methoxy, 2'-acetylamino, 2'-aminoethyl and 2'-O-methoxyethyl, and Base represents a nucleic acid base selected from A, G, C, T and U; preferably, the 5'-phosphate analog modified nucleotide has a vinyl phosphate modified nucleotide shown in formula (XIII), wherein R is selected from H, OH, fluorine, 2'-methoxy, 2'-acetylamino, 2'-aminoethyl and 2'-O-methoxyethyl; more preferably, the 5'-phosphonate analog modified nucleotide is APU shown in formula (XIV) or VPUm shown in formula (XV); Preferably, the oligonucleotide comprises a diol nucleic acid (GNA) modification, and the ribonucleotide has a diol nucleic acid structure as shown in formula (I), preferably an S-isomer; wherein Base represents a nucleic acid base selected from A, G, C and U; preferably, the oligonucleotide comprises Tgn, which is a thymidine-diol nucleic acid as shown in formula (II), preferably an S-isomer thereof; preferably, the oligonucleotide comprises Cgn, which is a cytidine-diol nucleic acid as shown in formula (III), preferably an S-isomer thereof; more preferably, the oligonucleotide comprises Agn, which is an adenosine-diol nucleic acid as shown in formula (IV), preferably an S-isomer thereof; preferably, the oligonucleotide comprises Ggn, which is a guanosine-diol nucleic acid as shown in formula (V), preferably an S-isomer thereof; Preferably, the oligonucleotide comprises a nucleoside-2'-phosphate represented by formula (VI), wherein Base represents a nucleic acid base selected from A, G, C and U; preferably, the oligonucleotide comprises U-2'5', which is a uridine-2'-phosphate represented by formula (VII); preferably, the oligonucleotide comprises C-2'5', which is a cytidine-2'-phosphate represented by formula (VIII); preferably, the oligonucleotide comprises A-2'5', which is an adenosine-2'-phosphate represented by formula (IX); preferably, the oligonucleotide comprises G-2'5', which is a guanosine-2'-phosphate represented by formula (X); Preferably, the oligonucleotide has a 5'-phosphate analog modified nucleotide at the 5' end; preferably, the 5'-phosphate analog modified nucleotide has a vinyl phosphonate modified nucleotide shown in formula (XI); preferably, the 5'-phosphate analog modified nucleotide has a vinyl phosphonate modified nucleotide shown in formula (XII); wherein R is selected from H, fluorine, 2'-methoxy, 2'-acetylamino, 2'-aminoethyl and 2'-O-methoxyethyl; Base represents a nucleic acid base selected from A, G, C and U; preferably, the oligonucleotide has APU at the 5' end, which is a 5'-phosphate analog modified uridylic acid (2'-acetylamino-5'-vinylphosphonate-uridylic acid) shown in formula (XIII); preferably, the oligonucleotide has VPU at the 5' end, which is a 5'-phosphate analog modified uridylic acid (2'-methoxy-5'-vinylphosphonate-uridylic acid) shown in formula (XIV); Preferably, the oligonucleotide comprises an inverted ribonucleotide (3'-3' linked nucleotide) as shown in formula (XV); Preferably, the oligonucleotide comprises invdA, which is an inverted deoxyadenylic acid as shown in formula (XVI); Preferably, the oligonucleotide comprises formula M, which is 2'-O-methyl-6-(3-(2-carboxyethyl)phenyl)-adenylic acid as shown in formula (XVII); 4. The oligonucleotide or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein: The oligonucleotide comprises at least one modified internucleotide linkage; Preferably, said at least one modified internucleotide bond is a phosphorothioate bond.

5. The oligonucleotide or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein: The sense strand is selected from SEQ ID NO. 8, 9, 38, 39, 40, 41, 55, 63, 64, 65, 66, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108 08, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 1 48, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188 88, 189, 190, 191, 192, 193, 194, 195, 196, 197, 735, 736, 737, 738, 739, 740, 741, 742, 743, 744, 745, 746, 747, 748, 749, 807, 808, 809, 810, 811, 812, 813, 814, 815, 816, 817, or any one of SEQ ID NO.405-419, 421-451, 453-556, 770-784, 829-839; the antisense strand is selected from SEQ ID NO. NO.258, 259, 265, 215, 273, 274, 275, 276, 278, 281, 283, 284, 286, 287, 297, 304, 306, 309, 316, 320, 326, 388, 389, 404, 750, 751, 752, 753, 754, 755, 756, 757, 758, 759, 760, 761, 762, 763, 764, 765, 766, 767, 768, 769, 818, 819, 820, 821, 822, 823, 824, 825, 826, 827, 828, an unmodified oligonucleotide as described in any one of 840-869, or SEQ ID NO.The modified oligonucleotide described in any one of 557-572, 574-598, 600-734, 785-806.

6. The oligonucleotide or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein: The oligonucleotide is selected from any one of the following sense strand and antisense strand combinations: (1) the sense strand comprises the sequence shown in SEQ ID NO.9, and the antisense strand comprises the sequence shown in SEQ ID NO.215; (2) the sense strand comprises the sequence shown in SEQ ID NO.38, and the antisense strand comprises the sequence shown in SEQ ID NO.258; (3) the sense strand comprises the sequence shown in SEQ ID NO.40, and the antisense strand comprises the sequence shown in SEQ ID NO.259; (4) the sense strand comprises the sequence shown in SEQ ID NO.55, and the antisense strand comprises the sequence shown in SEQ ID NO.265; (5) the sense strand comprises the sequence shown in SEQ ID NO.63, and the antisense strand comprises the sequence shown in SEQ ID NO.273; (6) the sense strand comprises the sequence shown in SEQ ID NO.64, and the antisense strand comprises the sequence shown in SEQ ID NO.274; (7) the sense strand comprises the sequence shown in SEQ ID NO.65, and the antisense strand comprises the sequence shown in SEQ ID NO.275; (8) the sense strand comprises the sequence shown in SEQ ID NO.66, and the antisense strand comprises the sequence shown in SEQ ID NO.276; (9) the sense strand comprises the sequence shown in SEQ ID NO.68, and the antisense strand comprises the sequence shown in SEQ ID NO.274; (10) the sense strand comprises the sequence shown in SEQ ID NO.74, and the antisense strand comprises the sequence shown in SEQ ID NO.278; (11) the sense strand comprises the sequence shown in SEQ ID NO.77, and the antisense strand comprises the sequence shown in SEQ ID NO.281; (12) the sense strand comprises the sequence shown in SEQ ID NO.79, and the antisense strand comprises the sequence shown in SEQ ID NO.283; (13) the sense strand comprises the sequence shown in SEQ ID NO.80, and the antisense strand comprises the sequence shown in SEQ ID NO.284; (14) the sense strand comprises the sequence shown in SEQ ID NO.80, and the antisense strand comprises the sequence shown in SEQ ID NO.767; (15) the sense strand comprises the sequence shown in SEQ ID NO.80, and the antisense strand comprises the sequence shown in SEQ ID NO.768; (16) the sense strand comprises the sequence shown in SEQ ID NO.80, and the antisense strand comprises the sequence shown in SEQ ID NO.769; (17) the sense strand comprises the sequence shown in SEQ ID NO.82, and the antisense strand comprises the sequence shown in SEQ ID NO.286; (18) the sense strand comprises the sequence shown in SEQ ID NO.83, and the antisense strand comprises the sequence shown in SEQ ID NO.287; (19) the sense strand comprises the sequence shown in SEQ ID NO.93, and the antisense strand comprises the sequence shown in SEQ ID NO.297; (20) the sense strand comprises the sequence shown in SEQ ID NO.99, and the antisense strand comprises the sequence shown in SEQ ID NO.304; (21) the sense strand comprises the sequence shown in SEQ ID NO.101, and the antisense strand comprises the sequence shown in SEQ ID NO.306; (22) the sense strand comprises the sequence shown in SEQ ID NO.104, and the antisense strand comprises the sequence shown in SEQ ID NO.309; (23) the sense strand comprises the sequence shown in SEQ ID NO.111, and the antisense strand comprises the sequence shown in SEQ ID NO.316; (24) the sense strand comprises the sequence shown in SEQ ID NO.115, and the antisense strand comprises the sequence shown in SEQ ID NO.320; (25) the sense strand comprises the sequence shown in SEQ ID NO.121, and the antisense strand comprises the sequence shown in SEQ ID NO.326; (26) the sense strand comprises the sequence shown in SEQ ID NO.182, and the antisense strand comprises the sequence shown in SEQ ID NO.388; (27) the sense strand comprises the sequence shown in SEQ ID NO.183, and the antisense strand comprises the sequence shown in SEQ ID NO.389; (28) the sense strand comprises the sequence shown in SEQ ID NO.197, and the antisense strand comprises the sequence shown in SEQ ID NO.404; (29) the sense strand comprises the sequence shown in SEQ ID NO.197, and the antisense strand comprises the sequence shown in SEQ ID NO.765; (30) the sense strand comprises the sequence shown in SEQ ID NO.197, and the antisense strand comprises the sequence shown in SEQ ID NO.766; (31) the sense strand comprises the sequence shown in SEQ ID NO.735, and the antisense strand comprises the sequence shown in SEQ ID NO.750; (32) the sense strand comprises the sequence shown in SEQ ID NO.736, and the antisense strand comprises the sequence shown in SEQ ID NO.751; (33) the sense strand comprises the sequence shown in SEQ ID NO.737, and the antisense strand comprises the sequence shown in SEQ ID NO.752; (34) the sense strand comprises the sequence shown in SEQ ID NO.738, and the antisense strand comprises the sequence shown in SEQ ID NO.753; (35) the sense strand comprises the sequence shown in SEQ ID NO.739, and the antisense strand comprises the sequence shown in SEQ ID NO.754; (36) the sense strand comprises the sequence shown in SEQ ID NO.740, and the antisense strand comprises the sequence shown in SEQ ID NO.755; (37) the sense strand comprises the sequence shown in SEQ ID NO.741, and the antisense strand comprises the sequence shown in SEQ ID NO.756; (38) the sense strand comprises the sequence shown in SEQ ID NO.742, and the antisense strand comprises the sequence shown in SEQ ID NO.757; (39) the sense strand comprises the sequence shown in SEQ ID NO.743, and the antisense strand comprises the sequence shown in SEQ ID NO.758; (40) the sense strand comprises the sequence shown in SEQ ID NO.744, and the antisense strand comprises the sequence shown in SEQ ID NO.759; (41) the sense strand comprises the sequence shown in SEQ ID NO.745, and the antisense strand comprises the sequence shown in SEQ ID NO.760; (42) the sense strand comprises the sequence shown in SEQ ID NO.746, and the antisense strand comprises the sequence shown in SEQ ID NO.761; (43) the sense strand comprises the sequence shown in SEQ ID NO.747, and the antisense strand comprises the sequence shown in SEQ ID NO.762; (44) the sense strand comprises the sequence shown in SEQ ID NO.748, and the antisense strand comprises the sequence shown in SEQ ID NO.763; (45) the sense strand comprises the sequence shown in SEQ ID NO.749, and the antisense strand comprises the sequence shown in SEQ ID NO.764; (46) the sense strand comprises the sequence shown in SEQ ID NO.807, and the antisense strand comprises the sequence shown in SEQ ID NO.818; (47) the sense strand comprises the sequence shown in SEQ ID NO.808, and the antisense strand comprises the sequence shown in SEQ ID NO.819; (48) the sense strand comprises the sequence shown in SEQ ID NO.809, and the antisense strand comprises the sequence shown in SEQ ID NO.820; (49) the sense strand comprises the sequence shown in SEQ ID NO.810, and the antisense strand comprises the sequence shown in SEQ ID NO.821; (50) the sense strand comprises the sequence shown in SEQ ID NO.811, and the antisense strand comprises the sequence shown in SEQ ID NO.822; (51) the sense strand comprises the sequence shown in SEQ ID NO.812, and the antisense strand comprises the sequence shown in SEQ ID NO.823; (52) the sense strand comprises the sequence shown in SEQ ID NO.813, and the antisense strand comprises the sequence shown in SEQ ID NO.824; (53) the sense strand comprises the sequence shown in SEQ ID NO.814, and the antisense strand comprises the sequence shown in SEQ ID NO.825; (54) the sense strand comprises the sequence shown in SEQ ID NO.815, and the antisense strand comprises the sequence shown in SEQ ID NO.826; (55) the sense strand comprises the sequence shown in SEQ ID NO.816, and the antisense strand comprises the sequence shown in SEQ ID NO.827; and (56) the sense strand comprises the sequence shown in SEQ ID NO.817, and the antisense strand comprises the sequence shown in SEQ ID NO.828; wherein each strand is independently 19 to 25 nucleotides in length.

7. The oligonucleotide or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein: The oligonucleotide comprises any one selected from the following sense strand and antisense strand combinations: (1) the sense strand comprises the sequence shown in SEQ ID NO.405, and the antisense strand comprises the sequence shown in SEQ ID NO.557; (2) the sense strand comprises the sequence shown in SEQ ID NO.406, and the antisense strand comprises the sequence shown in SEQ ID NO.558; (3) the sense strand comprises the sequence shown in SEQ ID NO.407, and the antisense strand comprises the sequence shown in SEQ ID NO.559; (4) the sense strand comprises the sequence shown in SEQ ID NO.408, and the antisense strand comprises the sequence shown in SEQ ID NO.560; (5) the sense strand comprises the sequence shown in SEQ ID NO.409, and the antisense strand comprises the sequence shown in SEQ ID NO.561; (6) the sense strand comprises the sequence shown in SEQ ID NO.410, and the antisense strand comprises the sequence shown in SEQ ID NO.562; (7) the sense strand comprises the sequence shown in SEQ ID NO.411, and the antisense strand comprises the sequence shown in SEQ ID NO.563; (8) the sense strand comprises the sequence shown in SEQ ID NO.412, and the antisense strand comprises the sequence shown in SEQ ID NO.564; (9) the sense strand comprises the sequence shown in SEQ ID NO.413, and the antisense strand comprises the sequence shown in SEQ ID NO.565; (10) the sense strand comprises the sequence shown in SEQ ID NO.414, and the antisense strand comprises the sequence shown in SEQ ID NO.566; (11) the sense strand comprises the sequence shown in SEQ ID NO.415, and the antisense strand comprises the sequence shown in SEQ ID NO.567; (12) the sense strand comprises the sequence shown in SEQ ID NO.416, and the antisense strand comprises the sequence shown in SEQ ID NO.568; (13) the sense strand comprises the sequence shown in SEQ ID NO.417, and the antisense strand comprises the sequence shown in SEQ ID NO.569; (14) the sense strand comprises the sequence shown in SEQ ID NO.418, and the antisense strand comprises the sequence shown in SEQ ID NO.570; (15) the sense strand comprises the sequence shown in SEQ ID NO.419, and the antisense strand comprises the sequence shown in SEQ ID NO.562; (16) the sense strand comprises the sequence shown in SEQ ID NO.419, and the antisense strand comprises the sequence shown in SEQ ID NO.571; (17) the sense strand comprises the sequence shown in SEQ ID NO.410, and the antisense strand comprises the sequence shown in SEQ ID NO.572; (18) the sense strand comprises the sequence shown in SEQ ID NO.421, and the antisense strand comprises the sequence shown in SEQ ID NO.562; (19) the sense strand comprises the sequence shown in SEQ ID NO.427, and the antisense strand comprises the sequence shown in SEQ ID NO.574; (20) the sense strand comprises the sequence shown in SEQ ID NO.430, and the antisense strand comprises the sequence shown in SEQ ID NO.577; (21) the sense strand comprises the sequence shown in SEQ ID NO.432, and the antisense strand comprises the sequence shown in SEQ ID NO.579; (22) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.580; (23) the sense strand comprises the sequence shown in SEQ ID NO.435, and the antisense strand comprises the sequence shown in SEQ ID NO.582; (24) the sense strand comprises the sequence shown in SEQ ID NO.436, and the antisense strand comprises the sequence shown in SEQ ID NO.583; (25) the sense strand comprises the sequence shown in SEQ ID NO.446, and the antisense strand comprises the sequence shown in SEQ ID NO.593; (26) the sense strand comprises the sequence shown in SEQ ID NO.453, and the antisense strand comprises the sequence shown in SEQ ID NO.600; (27) the sense strand comprises the sequence shown in SEQ ID NO.455, and the antisense strand comprises the sequence shown in SEQ ID NO.602; (28) the sense strand comprises the sequence shown in SEQ ID NO.458, and the antisense strand comprises the sequence shown in SEQ ID NO.605; (29) the sense strand comprises the sequence shown in SEQ ID NO.465, and the antisense strand comprises the sequence shown in SEQ ID NO.612; (30) the sense strand comprises the sequence shown in SEQ ID NO.469, and the antisense strand comprises the sequence shown in SEQ ID NO.616; (31) the sense strand comprises the sequence shown in SEQ ID NO.475, and the antisense strand comprises the sequence shown in SEQ ID NO.622; (32) the sense strand comprises the sequence shown in SEQ ID NO.537, and the antisense strand comprises the sequence shown in SEQ ID NO.685; (33) the sense strand comprises the sequence shown in SEQ ID NO.538, and the antisense strand comprises the sequence shown in SEQ ID NO.686; (34) the sense strand comprises the sequence shown in SEQ ID NO.419, and the antisense strand comprises the sequence shown in SEQ ID NO.700; (35) the sense strand comprises the sequence shown in SEQ ID NO.432, and the antisense strand comprises the sequence shown in SEQ ID NO.701; (36) the sense strand comprises the sequence shown in SEQ ID NO.419, and the antisense strand comprises the sequence shown in SEQ ID NO.702; (37) the sense strand comprises the sequence shown in SEQ ID NO.430, and the antisense strand comprises the sequence shown in SEQ ID NO.703; (38) the sense strand comprises the sequence shown in SEQ ID NO.432, and the antisense strand comprises the sequence shown in SEQ ID NO.704; (39) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.705; (40) the sense strand comprises the sequence shown in SEQ ID NO.435, and the antisense strand comprises the sequence shown in SEQ ID NO.706; (41) the sense strand comprises the sequence shown in SEQ ID NO.432, and the antisense strand comprises the sequence shown in SEQ ID NO.707; (42) the sense strand comprises the sequence shown in SEQ ID NO.553, and the antisense strand comprises the sequence shown in SEQ ID NO.710; (43) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.712; (44) the sense strand comprises the sequence shown in SEQ ID NO.553, and the antisense strand comprises the sequence shown in SEQ ID NO.716; (45) the sense strand comprises the sequence shown in SEQ ID NO.553, and the antisense strand comprises the sequence shown in SEQ ID NO.717; (46) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.725; (47) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.726; (48) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.727; (49) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.729; (50) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.733; (51) the sense strand comprises the sequence shown in SEQ ID NO.553, and the antisense strand comprises the sequence shown in SEQ ID NO.785; (52) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.786; (53) the sense strand comprises the sequence shown in SEQ ID NO.770, and the antisense strand comprises the sequence shown in SEQ ID NO.787; (54) the sense strand comprises the sequence shown in SEQ ID NO.771, and the antisense strand comprises the sequence shown in SEQ ID NO.788; (55) the sense strand comprises the sequence shown in SEQ ID NO.772, and the antisense strand comprises the sequence shown in SEQ ID NO.789; (56) the sense strand comprises the sequence shown in SEQ ID NO.773, and the antisense strand comprises the sequence shown in SEQ ID NO.790; (57) the sense strand comprises the sequence shown in SEQ ID NO.774, and the antisense strand comprises the sequence shown in SEQ ID NO.791; (58) the sense strand comprises the sequence shown in SEQ ID NO.775, and the antisense strand comprises the sequence shown in SEQ ID NO.792; (59) the sense strand comprises the sequence shown in SEQ ID NO.776, and the antisense strand comprises the sequence shown in SEQ ID NO.793; (60) the sense strand comprises the sequence shown in SEQ ID NO.777, and the antisense strand comprises the sequence shown in SEQ ID NO.794; (61) the sense strand comprises the sequence shown in SEQ ID NO.778, and the antisense strand comprises the sequence shown in SEQ ID NO.795; (62) the sense strand comprises the sequence shown in SEQ ID NO.779, and the antisense strand comprises the sequence shown in SEQ ID NO.796; (63) the sense strand comprises the sequence shown in SEQ ID NO.780, and the antisense strand comprises the sequence shown in SEQ ID NO.797; (64) the sense strand comprises the sequence shown in SEQ ID NO.781, and the antisense strand comprises the sequence shown in SEQ ID NO.798; (65) the sense strand comprises the sequence shown in SEQ ID NO.782, and the antisense strand comprises the sequence shown in SEQ ID NO.799; (66) the sense strand comprises the sequence shown in SEQ ID NO.783, and the antisense strand comprises the sequence shown in SEQ ID NO.800; (67) the sense strand comprises the sequence shown in SEQ ID NO.784, and the antisense strand comprises the sequence shown in SEQ ID NO.801; (68) the sense strand comprises the sequence shown in SEQ ID NO.553, and the antisense strand comprises the sequence shown in SEQ ID NO.802; (69) the sense strand comprises the sequence shown in SEQ ID NO.553, and the antisense strand comprises the sequence shown in SEQ ID NO.803; (70) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.804; (71) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.805; (72) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.806; (73) the sense strand comprises the sequence shown in SEQ ID NO.772, and the antisense strand comprises the sequence shown in SEQ ID NO.841; (74) the sense strand comprises the sequence shown in SEQ ID NO.775, and the antisense strand comprises the sequence shown in SEQ ID NO.842; (75) the sense strand comprises the sequence shown in SEQ ID NO.776, and the antisense strand comprises the sequence shown in SEQ ID NO.843; (76) the sense strand comprises the sequence shown in SEQ ID NO.777, and the antisense strand comprises the sequence shown in SEQ ID NO.844; (77) the sense strand comprises the sequence shown in SEQ ID NO.783, and the antisense strand comprises the sequence shown in SEQ ID NO.848; (78) the sense strand comprises the sequence shown in SEQ ID NO.783, and the antisense strand comprises the sequence shown in SEQ ID NO.854; (79) the sense strand comprises the sequence shown in SEQ ID NO.784, and the antisense strand comprises the sequence shown in SEQ ID NO.849; (80) the sense strand comprises the sequence shown in SEQ ID NO.831, and the antisense strand comprises the sequence shown in SEQ ID NO.858; (81) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.869; (82) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.840; (83) the sense strand comprises the sequence shown in SEQ ID NO.772, and the antisense strand comprises the sequence shown in SEQ ID NO.841; (84) the sense strand comprises the sequence shown in SEQ ID NO.775, and the antisense strand comprises the sequence shown in SEQ ID NO.842; (85) the sense strand comprises the sequence shown in SEQ ID NO.776, and the antisense strand comprises the sequence shown in SEQ ID NO.843; (86) the sense strand comprises the sequence shown in SEQ ID NO.777, and the antisense strand comprises the sequence shown in SEQ ID NO.844; (87) the sense strand comprises the sequence shown in SEQ ID NO.783, and the antisense strand comprises the sequence shown in SEQ ID NO.848; (88) the sense strand comprises the sequence shown in SEQ ID NO.783, and the antisense strand comprises the sequence shown in SEQ ID NO.854; (89) the sense strand comprises the sequence shown in SEQ ID NO.784, and the antisense strand comprises the sequence shown in SEQ ID NO.849; (90) the sense strand comprises the sequence shown in SEQ ID NO.831, and the antisense strand comprises the sequence shown in SEQ ID NO.858; and (91) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.869; Preferably, the oligonucleotide comprises any one selected from the following sense strand and antisense strand combinations: (1) the sense strand comprises the sequence shown in SEQ ID NO.406, and the antisense strand comprises the sequence shown in SEQ ID NO.558; (2) the sense strand comprises the sequence shown in SEQ ID NO.407, and the antisense strand comprises the sequence shown in SEQ ID NO.559; (3) the sense strand comprises the sequence shown in SEQ ID NO.409, and the antisense strand comprises the sequence shown in SEQ ID NO.561; (4) the sense strand comprises the sequence shown in SEQ ID NO.410, and the antisense strand comprises the sequence shown in SEQ ID NO.562; (5) the sense strand comprises the sequence shown in SEQ ID NO.411, and the antisense strand comprises the sequence shown in SEQ ID NO.563; (6) the sense strand comprises the sequence shown in SEQ ID NO.419, and the antisense strand comprises the sequence shown in SEQ ID NO.702; (7) the sense strand comprises the sequence shown in SEQ ID NO.430, and the antisense strand comprises the sequence shown in SEQ ID NO.703; (8) the sense strand comprises the sequence shown in SEQ ID NO.432, and the antisense strand comprises the sequence shown in SEQ ID NO.704; (9) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.705; (10) the sense strand comprises the sequence shown in SEQ ID NO.435, and the antisense strand comprises the sequence shown in SEQ ID NO.706; (11) the sense strand comprises the sequence shown in SEQ ID NO.432, and the antisense strand comprises the sequence shown in SEQ ID NO.707; (12) the sense strand comprises the sequence shown in SEQ ID NO.553, and the antisense strand comprises the sequence shown in SEQ ID NO.710; (13) the sense strand comprises the sequence shown in SEQ ID NO.553, and the antisense strand comprises the sequence shown in SEQ ID NO.716; (14) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.712; (15) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.727; (16) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.729; (17) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.733; (18) the sense strand comprises the sequence shown in SEQ ID NO.553, and the antisense strand comprises the sequence shown in SEQ ID NO.785; (19) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.786; (20) the sense strand comprises the sequence shown in SEQ ID NO.453, and the antisense strand comprises the sequence shown in SEQ ID NO.600; (21) the sense strand comprises the sequence shown in SEQ ID NO.537, and the antisense strand comprises the sequence shown in SEQ ID NO.685; (22) the sense strand comprises the sequence shown in SEQ ID NO.538, and the antisense strand comprises the sequence shown in SEQ ID NO.686; (23) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.840; (24) the sense strand comprises the sequence shown in SEQ ID NO.772, and the antisense strand comprises the sequence shown in SEQ ID NO.841; (25) the sense strand comprises the sequence shown in SEQ ID NO.775, and the antisense strand comprises the sequence shown in SEQ ID NO.842; (26) the sense strand comprises the sequence shown in SEQ ID NO.776, and the antisense strand comprises the sequence shown in SEQ ID NO.843; (27) the sense strand comprises the sequence shown in SEQ ID NO.777, and the antisense strand comprises the sequence shown in SEQ ID NO.844; (28) the sense strand comprises the sequence shown in SEQ ID NO.783, and the antisense strand comprises the sequence shown in SEQ ID NO.848; (29) the sense strand comprises the sequence shown in SEQ ID NO.783, and the antisense strand comprises the sequence shown in SEQ ID NO.854; (30) the sense strand comprises the sequence shown in SEQ ID NO.784, and the antisense strand comprises the sequence shown in SEQ ID NO.849; (31) the sense strand comprises the sequence shown in SEQ ID NO.831, and the antisense strand comprises the sequence shown in SEQ ID NO.858; and (32) the sense strand comprises the sequence shown in SEQ ID NO.433, and the antisense strand comprises the sequence shown in SEQ ID NO.869; wherein each strand is independently 19 to 25 nucleotides in length.

8. A conjugate for reducing lipoprotein a (Lp(a)) gene expression or a pharmaceutically acceptable salt thereof, comprising: (i) the oligonucleotide or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, and (ii) a ligand conjugated to the oligonucleotide or a pharmaceutically acceptable salt thereof, wherein at least one nucleotide of the oligonucleotide is conjugated to a targeting ligand; Preferably, the targeting ligand comprises a carbohydrate, an amino sugar, cholesterol, a polypeptide or a lipid; Preferably, the targeting ligand comprises an N-acetylgalactosamine (GalNAc) moiety; Preferably, the GalNac moiety is a monovalent GalNAc moiety, a divalent GalNAc moiety, a trivalent GalNAc moiety or a tetravalent GalNAc moiety; Preferably, the targeting ligand is selected from L96, A1, A3 and NAG25s; 9. A composition comprising the oligonucleotide or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7 or the conjugate or a pharmaceutically acceptable salt thereof according to claim 8, and optionally a pharmaceutically acceptable carrier; Preferably, the composition is in the form of an oral agent, an intravenous injection, a subcutaneous injection or an intramuscular injection; Preferably, the combination further comprises other drugs for treating and / or preventing Lp(a)-related diseases; Preferably, the Lp(a)-related disease is selected from Berger'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, cerebrovascular atherosclerosis, cerebrovascular disease, venous thrombosis, stroke, atherosclerosis, thrombosis, coronary heart disease or aortic stenosis and / or any other disease or pathology associated with elevated levels of Lp(a)-containing particles.

10. Use of the oligonucleotide according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof or the composition according to claim 9 in the preparation of a medicament for treating and / or preventing Lp(a)-related diseases; Preferably, the Lp(a)-related disease is selected from Berger'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, cerebrovascular atherosclerosis, cerebrovascular disease, venous thrombosis, stroke, atherosclerosis, thrombosis, coronary heart disease or aortic stenosis and / or any other disease or pathology associated with elevated levels of Lp(a)-containing particles.

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