Oligonucleotides targeting type 13 17β-hydroxysteroid dehydrogenase and their applications

Abstract

This application provides an oligonucleotide targeting type 13 17β-hydroxysteroid dehydrogenase (HSD17B13) and its use therein, wherein the targeting oligonucleotide can inhibit the expression level of the HSD17B13 gene.

Description

Oligonucleotides targeting type 13 17β-hydroxysteroid dehydrogenase and uses thereof Technical Field

[0001] The present disclosure relates to an oligonucleotide, in particular to an oligonucleotide for inhibiting 17β-hydroxysteroid dehydrogenase (HSD17Bs) gene expression and treating HSD17B13-related diseases. Background Art

[0002] 17β-Hydroxysteroid dehydrogenases (HSD17Bs) are a group of enzymes that catalyze the conversion between 17-keto- and 17-hydroxysteroids. To date, 15 members of the HSD17Bs family have been discovered, most of which are involved in regulating the biological activity of sex hormones, including HSD17B1, HSD17B2, HSD17B3, HSD17B5, and HSD17B6. Other family members also participate in fatty acid metabolism, cholesterol biosynthesis, and bile acid production in the body (Saloniemi, T., Jokela, H., Strauss, L., Pakarinen, P., Poutanen, M. The diversity of sex steroid action: novel functions of hydroxysteroid (17beta) dehydrogenases as revealed by genetically modified mouse models. J. Endocrinol. 2012.). A splice variant (rs72613567:TA) in HSD17B13, encoding the hepatic lipid droplet protein hydroxysteroid 17-β dehydrogenase 13, was found to be associated with reduced levels of aspartate aminotransferase (ALT) and alanine aminotransferase (AST). HSD17B13 was also associated with a lower risk of hepatocellular carcinoma (P=0.047).

[0003] HSD17B13, originally named SCDR9, was first cloned from a human liver cDNA library in 2007 (Liu, S., Huang, C., Li, D., Ren, W., Zhang, H., Qi, M., Li, X., Yu, L., Molecular cloning and expression analysis of a new gene for short-chain dehydrogenase / reductase 9. Acta Biochim. Pol, 2007). The human HSD17B13 gene is located on chromosome 4q22.1 and shares a high degree of sequence homology (78%) with HSD17B11 and its protein, which also maps to the same chromosome. Similar to other 17BHSD members, HSD17B13 contains two conserved motifs: a TGXGXXXG motif associated with NAD(P)(H) binding and a YXXXK motif important for its catalytic activity.

[0004] The role of HSD17B13 in liver physiology

[0005] In 2008, Horiguchi compared the localization of overexpressed HSD17B11 and HSD17B13 and found that HSD17B13 expression is localized to the surface of lipid droplets (LDs). He also found that the N-terminal 35 amino acids of HSD17B13 play a major role in its localization to LDs. Horiguchi identified HSD17B13 as a new LD-associated protein, which is primarily expressed in the liver (Straub, BK, Stoeffel, P., Heid, H., Zimbelmann, R., Schirmacher, P., Differential pattern of lipid droplet-associated proteins and de novo perilipin expression in hepatocyte steatogenesis. Hepatology, 2008). Next, the researchers expressed human HSD17B13 in HepG2 and Huh7 liver cancer cells, clearly demonstrating its localization to the surface of LDs. Immunohistochemical studies of human fatty liver samples further supported the finding that HSD17B13 is an LD-associated protein. Subsequent researchers, using comparative proteomics strategies, discovered that HSD17B13 is located on the surface of hepatic lipid droplets and that its expression is significantly upregulated in the livers of patients with nonalcoholic fatty liver disease (NAFLD) and NAFLD mice (Abul-Husn, NS, Cheng, X., Li, AH, Xin, Y., Schurmann, C., Stevis, P., Liu, Y., Kozlitina, J., Stender, S., Wood, GC, et al., A protein-truncating HSD17B13 variant and protection from chronic liver disease. N. Engl. J. Med., 2018). Accumulating evidence suggests that HSD17B13 plays a key role in hepatic lipid metabolism and that its abnormal expression contributes to the further development of NAFLD.

[0006] Although HSD17B13 has been identified as a liver-specific LD-associated protein and implicated in the pathogenesis of NAFLD, the physiological functions of HSD17B13 remain largely unknown. Based on the high similarity in protein sequence and subcellular localization between HSD17B11 and HSD17B13, it has been previously hypothesized that HSD17B13 may also be involved in sex hormone metabolism. Several studies have compared HSD17B13 with HSD17B11, but have found that HSD17B13 is more likely to be involved in fatty acid metabolism, suggesting that the distinct sequence regions between HSD17B13 and HSD17B11 may be the primary domains responsible for HSD17B13's function and enzymatic activity.

[0007] Studies have shown that overexpression of human HSD17B13 increases the number and size of LDs in cultured hepatocytes. Loss-of-function variants in the HSD17B13 gene can protect against chronic liver damage and mitigate the progression of NAFLD in European Americans. Poutanen and Foti's groups recently reported several important phenotypes of HSD17B13 knockout (KO) mice. No changes in serum sex steroid concentrations were found in HSD17B13 KO mice, suggesting that the enzyme may have no effect on sex hormone metabolism. Interestingly, histological analysis showed that HSD17B13KO mice exhibited an aging-associated hepatic steatosis phenotype (Adam, M., Heikela, H., Sobolewski, C., Portius, D., Maki-Jouppila, J., Mehmood, A., Adhikari, P., Esposito, I., Elo, LL, Zhang, FP, et al., Hydroxysteroid (17beta) dehydrogenase 13 deficiency triggers hepatic steatosis and in flammation in mice. Faseb. J., 2018).

[0008] To date, the physiological function of HSD17B13 remains largely undefined. The fact that HSD17B13 is selectively expressed in the liver and on the surface of lipid droplets suggests that it may play a key role in lipid droplet function and be a key player in hepatic lipid homeostasis. Together, these findings suggest that HSD17B13 plays an important role in hepatic lipid metabolism.

[0009] Association of HSD17B13 with liver disease

[0010] Over the past decade, researchers have focused on the role of LD-associated proteins in the progression of NAFLD. Among the numerous LD-associated proteins, the PAT family, including PLIN1–5, is the most extensively studied. High expression of PLIN1 and PLIN2 has been shown to significantly increase the progression of NAFLD and NASH in liver patients (Carr, RM, et al., 2014).

[0011] Through a comprehensive analysis of the differential expression of LD-associated proteins and human NAFLD proteins, the researchers found that 54 LD-associated proteins in the human body have an upregulating effect on NAFLD, and 35 LD-associated proteins have a downregulating effect on NAFLD. An important discovery is that a human liver-specific LD-associated protein, HSD17B13, plays an important role in the pathological process of NAFLD. Overexpression of HSD17B13 in human HCC cell lines leads to excessive lipid accumulation. At the same time, overexpression of HSD17B13 in the liver of C57BL / 6 mice increases lipogenesis and TG content in the liver. It was further observed that hepatic overexpression of HSD17B13 was accompanied by higher hepatic sterol regulatory element binding protein 1 (SREBP-1) maturation and fatty acid synthase (FAS) protein expression. SREBP1-dependent de novo lipogenesis (DNL) in the liver may contribute to the HSD17B13-induced fatty liver phenotype (Straub, BK, Stoeffel, P., Heid, H., Zimbelmann, R., Schirmacher, P., Differential pattern of lipid droplet-associated proteins and de novo perilipin expression in he-patocyte steatogenesis. Hepatology, 2008. Su, W., et al. Role of HSD17B13 in the liver physiology and pathophysiology. Molecular and Cellular Endocrinology, 2019). Another independent study also revealed the relationship between HSD17B13 and NAFLD. In this study, the HSD17B13 gene is involved in the development of non-alcoholic steatohepatitis (NASH), cirrhosis, and HCC (hepatocellular carcinoma), and is upregulated in patients with NASH and fatty liver (Kampf, C., Mardinoglu, A., Fagerberg, L., Hallstrom, BM, Edlund, K., Lundberg, E., Ponten, F., Nielsen, J., Uhlen, M., The human liver-specific proteome defined by transcriptomics and antibody-based profiling. Faseb. J., 2014).HSD17B13 is not only a direct target of SREBP-1c but also a potential mediator that promotes SREBP-1c maturation. Therefore, it is speculated that the vicious cycle between SREBP-1c and HSD17B13 may play an important role in the pathogenesis of NAFLD (Figure 1).

[0012] Liver LDs are highly active organelles, where HSD17B13 plays an important role in maintaining their homeostasis. HSD17B13 is a direct target gene of SREBP-1c and is involved in de novo hepatic lipogenesis. Insulin, saturated fatty acids (FAs), and LXR agonists, such as T0901317, induce SREBP-1c expression and maturation. SREBP-1c transcriptionally increases HSD17B13 expression, which further promotes SREBP1c maturation, thereby forming a positive feedback loop to enhance lipogenesis. Overexpression of HSD17B13 leads to lipid synthesis and LD enlargement, ultimately causing hepatic steatosis. When HSD17B13 is knocked out, the long-term accumulation of HSD17B13 substrates or the lack of metabolites may disrupt lipid homeostasis and contribute to age-related steatosis and inflammation.

[0013] Currently, there is no treatment for chronic fibrotic liver disease. The standard of care for individuals with chronic fibrotic liver disease includes lifestyle modifications and management of associated comorbidities such as hypertension, hyperlipidemia, diabetes, and obesity. Accordingly, as the prevalence of chronic fibrotic liver disease has increased over the past decade and is expected to continue to increase, there is a need for alternative treatments for individuals with chronic fibrotic liver disease.

[0014] As research into the pathogenesis of NAFLD / NASH continues to deepen, it has been discovered that pathogenic pathways such as insulin resistance, lipotoxicity, oxidative stress, immune or cytokine or mitochondrial function changes, and apoptosis all contribute to the occurrence and development of NASH. Therefore, targeted interventions targeting these pathogenesis mechanisms are a hot topic in current research. The exploration of these therapeutic targets is currently at different stages of research, focusing on interventions targeting different targets in the anti-fibrotic, metabolic, and inflammatory categories. Combined multi-drug interventions targeting different targets in the pathogenesis of NASH may be an important direction for future research. Therapeutic targets need to take into account the prevention and treatment of hepatic steatosis, inflammation, hepatocellular damage, and fibrosis. Summary of the Invention

[0015] The purpose of the present disclosure is to provide an inhibitor targeting type 13 17β hydroxysteroid dehydrogenase (HSD17B13) with good efficacy, high safety and long-lasting efficacy.

[0016] The present disclosure provides double-stranded ribonucleic acids (dsRNAs) and methods of using the dsRNAs to inhibit expression of the HSD17B13 gene in cells or mammals, wherein the dsRNA targets the HSD17B13 gene. Also provided herein are compositions and methods for treating pathological conditions and diseases in mammals caused by HSD17B13 expression. dsRNAs direct the sequence-specific degradation of mRNA through a process known as RNA interference (RNAi).

[0017] In one aspect, the present disclosure provides an oligonucleotide or a pharmaceutically acceptable salt thereof for reducing HSD17B13 gene expression, wherein the oligonucleotide comprises a sense strand and an antisense strand, wherein the sense strand has a sequence identity of at least 80% or more to a sequence as shown in any one of SEQ ID NOs. 1-26, 28-40, and 42-97, or a fragment thereof, or a modified sequence thereof, preferably a sequence identity of 85%, 90%, 95%, 96%, 97%, 98%, or 99% or more; and the antisense strand has a sequence identity of at least 80% or more to a sequence as shown in any one of SEQ ID NOs. 99-124, 126-138, and 140-195, or a fragment thereof, or a modified sequence thereof, preferably a sequence identity of 85%, 90%, 95%, 96%, 97%, 98%, or 99% or more.

[0018] In another aspect, the present disclosure provides a conjugate for reducing HSD17B13 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 aforementioned oligonucleotide is conjugated to a targeting ligand.

[0019] 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.

[0020] 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 HSD17B13-related diseases.

[0021] In another aspect, the present disclosure provides a method for inhibiting HSD17B13 gene expression in a subject, comprising administering to the subject the aforementioned oligonucleotide or a pharmaceutically acceptable salt thereof, or the aforementioned conjugate or a pharmaceutically acceptable salt thereof, or the aforementioned composition.

[0022] In another aspect, the present disclosure provides a method for preventing or treating a subject suffering from an HSD17B13-related disease, comprising administering to the subject the aforementioned oligonucleotide or a pharmaceutically acceptable salt thereof, the aforementioned conjugate or a pharmaceutically acceptable salt thereof, or the aforementioned composition.

[0023] Experiments have shown that the candidate compounds disclosed herein can effectively reduce the level of HSD17B13 in the body and are effective inhibitors of HSD17B13. The present disclosure provides a method for treating a disease or condition associated with the HSD17B13 protein, comprising administering an effective dose of a dsRNA agent disclosed herein to a subject to inhibit the expression of the HSD17B13 gene. In certain aspects, the HSD17B13-related disease or condition is chronic fibrotic liver disease, wherein the aforementioned chronic fibrotic liver disease includes any other disease or pathology of NAFLD and NASH. In some embodiments, the method further comprises: co-administering an additional non-HSD17B13 siRNA therapeutic agent to the subject, wherein the non-HSD17B13 siRNA therapeutic agent includes one or more of the following: glitazones, vitamin E, liraglutide, metformin, statins, oxoxifylline, and obeticholic acid. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] FIG1 shows the mechanism by which HSD17B13 participates in hepatic LD homeostasis.

[0025] FIG2 shows the solid phase synthesis process of siRNA. DETAILED DESCRIPTION

[0026] 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.

[0027] As used herein, the term "approximately" or "approximately" as applied to one or more target values ​​refers to a value similar to a 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).

[0028] Described herein are RNAi agents targeting HSD17B13 for selectively and effectively inhibiting the expression of the HSD17B13 gene. HSD17B13 is the name of the gene encoding type 13 17β-hydroxysteroid dehydrogenase. The RNAi agents targeting HSD17B13 described herein can be used to prevent or treat the following diseases or to prepare medicaments for preventing or treating the following diseases, including but not limited to: non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis and cirrhosis, and complications associated with these diseases, such as type 2 diabetes, cardiovascular disease, and chronic kidney disease.

[0029] As used herein, "target sequence" refers to a continuous portion of the nucleotide sequence of an mRNA molecule formed during the transcription of the HSD17B13 gene, including mRNA that is an RNA processing product of the primary transcription product.

[0030] 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.

[0031] 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.

[0032] 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.

[0033] 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 a siRNA oligonucleotide.

[0034] 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.

[0035] 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 chains with "sense" and "antisense" orientations relative to the target RNA (i.e., the HSD17B13 gene). In some embodiments of the present disclosure, double-stranded RNA (dsRNA) triggers the degradation of the target RNA (e.g., mRNA) by a post-transcriptional gene silencing mechanism referred to herein as RNA interference or RNAi. In general, the majority of the nucleotides of each chain of the dsRNA molecule are ribonucleotides, but as described in detail herein, each chain or both chains may also include one or more non-ribonucleotides, such as deoxyribonucleotides or modified nucleotides. In addition, as used herein, "iRNA" may include ribonucleotides with chemical modifications; iRNA may include substantial modifications at multiple nucleotides.

[0036] As used herein, the terms "iRNA," "RNAi agent," "iRNA agent," and "RNA interference agent" are used interchangeably herein and refer to agents that comprise RNA, as such terms are defined herein, and mediate 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, expression of HSD17B13 in cells, for example, cells within an individual, such as a mammalian individual.

[0037] 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, the first siRNA conjugate or the second siRNA conjugate, or the siRNA sense strand conjugate or the siRNA antisense strand conjugate, depending on the context.

[0038] 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.

[0039] 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.

[0040] As used herein, the term "naked sequence" refers to an unmodified nucleotide sequence.

[0041] 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.

[0042] The phrase "inhibiting the expression of HSD17B13" is intended to refer to inhibiting the expression of any HSD17B13 gene (such as, for example, a mouse HSD17B13 gene, a rat HSD17B13 gene, a monkey HSD17B13 gene, or a human HSD17B13 gene), as well as variants or mutants of HSD17B13 genes. Thus, in the context of genetically manipulated cells, cell populations, or organisms, the HSD17B13 gene can be a wild-type HSD17B13 gene, a mutant HSD17B13 gene, or a transgenic HSD17B13 gene.

[0043] "Inhibiting the expression of the HSD17B13 gene" includes any level of inhibition of the HSD17B13 gene, for example, at least partial inhibition of the expression of the HSD17B13 gene. The expression of the HSD17B13 gene can be assessed based on the level or change in the level of any variable associated with the expression of the HSD17B13 gene, for example, the level of HSD17B13 mRNA or the level of HSD17B13 protein, or indirectly reflects the inhibition of the level of HSD17B13 protein by inhibiting the mRNA levels of Gluc and HSD17B13 fusion protein genes and thereby inhibiting the level of Gluc protein.

[0044] Inhibition can be assessed by a decrease in the absolute or relative level of one or more variables associated with HSD17B13 expression compared to a control level. The control level can be any type of control level used in the art, for example, a pre-dose baseline level, or a level determined from a similar subject that is untreated or treated with a control such as, for example, a buffer-only control or an inactive agent control.

[0045] 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.

[0046] 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.

[0047] 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 HSD17B13-related disorder in a subject. Treatment also encompasses reducing one or more signs or symptoms associated with undesirable HSD17B13 expression; lessening the extent of undesirable HSD17B13 activation or stabilization; ameliorating or alleviating undesirable HSD17B13 activation or stabilization. Treatment also encompasses reducing one or more signs or symptoms associated with undesirable HSD17B13 expression. "Treatment" may also mean prolonging survival compared to expected survival in the absence of treatment.

[0048] As used herein, the terms "prevention" or "preventing" when used in reference to a disease or condition that would benefit from a decrease in HSD17B13 gene expression or HSD17B13 protein production.

[0049] As used herein, the term "therapeutically effective amount" is intended to encompass an amount of an RNAi agent that, when administered to a subject with an HSD17B13-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.

[0050] As used herein, the term "prophylactically effective amount" is intended to include an amount of an RNAi agent that, when administered to a subject with an HSD17B13-related disorder, is sufficient to prevent or ameliorate the disorder or one or more symptoms of the disorder. Amelioration of a disease includes slowing the progression of the disease or reducing the severity of a later-developing disease. 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.

[0051] In one aspect, the present disclosure provides an oligonucleotide or a pharmaceutically acceptable salt thereof for reducing HSD17B13 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. 1-26, 28-40, and 42-97, or a fragment thereof, or a modified sequence 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. 99-124, 126-138, and 140-195, or a fragment thereof, or a modified sequence thereof, preferably a sequence having 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity.

[0052] In some embodiments, wherein each strand is independently 19 to 25 nucleotides in length.

[0053] In some embodiments, the antisense strand is 19 to 23 nucleotides in length.

[0054] In some embodiments, the sense strand is 19 to 23 nucleotides in length.

[0055] 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.

[0056] In some embodiments, the antisense strand has one or two overhangs.

[0057] In some embodiments, the sense strand has one or two overhangs.

[0058] In some embodiments, the oligonucleotide comprises a 3'-overhang sequence that is 1 or 2 nucleotides in length.

[0059] In some embodiments, the oligonucleotide comprises a 5'-overhang sequence that is 1 or 2 nucleotides in length.

[0060] 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, AC, AG, AU, CA, CC, CU, GA, GC, GG, GU, UA, UC, UG, UU.

[0061] In some embodiments, the oligonucleotide comprises an antisense strand and a sense strand each ranging from 19 to 23 nucleotides in length.

[0062] In some embodiments, the sense strand and the antisense strand form a duplex region.

[0063] 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.

[0064] 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.

[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 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.

[0066] 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.

[0067] 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.

[0068] 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.

[0069] 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.

[0070] 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.

[0071] In some embodiments, the oligonucleotide or its salt may be in the form of a hydrate or a solvate.

[0072] In some embodiments, the oligonucleotide comprises at least one modified nucleotide.

[0073] In some embodiments, the oligonucleotide comprises at least one 2'-modified nucleotide.

[0074] 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.

[0075] 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;

[0076] 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; preferably, the oligonucleotide comprises Tgn, which is a thymidine-diol nucleic acid as shown in formula (II), preferably an S-isomer; preferably, the oligonucleotide comprises Cgn, which is a cytidine-diol nucleic acid as shown in formula (III), preferably an S-isomer; preferably, the oligonucleotide comprises Agn, which is an adenosine-diol nucleic acid as shown in formula (IV), preferably an S-isomer; preferably, the oligonucleotide comprises Ggn, which is a guanosine-diol nucleic acid as shown in formula (V), preferably an S-isomer;

[0077] In some embodiments, the oligonucleotide comprises a nucleoside-2'-phosphate of formula (VI); preferably, the oligonucleotide comprises U-2'5', which is a uridine-2'-phosphate of formula (VII); preferably, the oligonucleotide comprises C-2'5', which is a cytidine-2'-phosphate of formula (VIII); preferably, the oligonucleotide comprises A-2'5', which is an adenosine-2'-phosphate of formula (IX); preferably, the oligonucleotide comprises G-2'5', which is a guanosine-2'-phosphate of formula (X);

[0078] Nucleoside-2'-phosphate

[0079] In some embodiments, the oligonucleotide has a 5'-phosphate analog modified nucleotide at the 5' terminus; preferably, the 5'-phosphate analog modified nucleotide has a vinyl phosphonate modified nucleotide of formula (XI); preferably, the 5'-phosphate analog modified nucleotide has a vinyl phosphonate modified nucleotide of formula (XII); preferably, the oligonucleotide has APU at the 5' terminus, which is a 5'-phosphate analog modified uridylic acid (2'-acetamido-5'-vinylphosphonate-uridylic acid) of formula (XIII); preferably, the oligonucleotide has VPU at the 5' terminus, which is a 5'-phosphate analog modified uridylic acid (2'-methoxy-5'-vinylphosphonate-uridylic acid) of formula (XIV);

[0080] In some embodiments, the oligonucleotide comprises a 6-(3-(2-carboxyethyl)phenyl)purine modified nucleotide; preferably, the oligonucleotide comprises formula M, which is a 2'-O-methyl-6-(3-(2-carboxyethyl)phenyl)-purine nucleotide shown in formula (XVII);

[0081] In some embodiments, the oligonucleotide comprises at least one modified internucleotide linkage.

[0082] 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.

[0083] In some embodiments, the sense strand comprises a sequence selected from any one of SEQ ID NO. 1-2, 4, 6-8, 10-11, 13-16, 22-23, 25, 31, 33-34, 36-40, 42-46, 52-56, 58-63, 65, 67, 72-80, 83-87, 90, 93-95, or a modified sequence thereof; the antisense strand comprises a sequence selected from any one of SEQ ID NO. 174, 175, 176, 177, 178, 181, 182, 183, 184, 185, 188, 191, 192, or any of the foregoing sequences.

[0084] In some embodiments, the sense strand comprises a sequence selected from any one of SEQ ID NO. 7, 33, 34, 38, 43, 52, 53, 59, 60, 61, 62, 63, 65, 67, 72, 73, 74, 75, 77, 78, 79, 83, 87, 90, and 94, or a modified sequence thereof; the antisense strand comprises a sequence selected from any one of SEQ ID NO. 105, 131, 132, 136, 141, 150, 151, 157, 158, 159, 160, 161, 163, 165, 170, 171, 172, 173, 175, 176, 177, 181, 185, 188, and 192, or a modified sequence thereof.

[0085] In some embodiments, the modified sequence of the sense strand comprises a sequence selected from any one of SEQ ID NOs. 198-212, 214-221, 223-258; and the modified sequence of the antisense strand comprises a sequence selected from any one of SEQ ID NOs. 261-275, 277-284, 286-321.

[0086] In some embodiments, the oligonucleotide is selected from any of the following sense and antisense strand combinations:

[0087] (1) the sense strand comprises the sequence shown in SEQ ID NO. 1, and the antisense strand comprises the sequence shown in SEQ ID NO. 99;

[0088] (2) the sense strand comprises the sequence shown in SEQ ID NO. 2, and the antisense strand comprises the sequence shown in SEQ ID NO. 100;

[0089] (3) the sense strand comprises the sequence shown in SEQ ID NO. 4, and the antisense strand comprises the sequence shown in SEQ ID NO. 102;

[0090] (4) the sense strand comprises the sequence shown in SEQ ID NO. 6, and the antisense strand comprises the sequence shown in SEQ ID NO. 104;

[0091] (5) the sense strand comprises the sequence shown in SEQ ID NO. 7, and the antisense strand comprises the sequence shown in SEQ ID NO. 105;

[0092] (6) the sense strand comprises the sequence shown in SEQ ID NO. 8, and the antisense strand comprises the sequence shown in SEQ ID NO. 106;

[0093] (7) the sense strand comprises the sequence shown in SEQ ID NO. 10, and the antisense strand comprises the sequence shown in SEQ ID NO. 108;

[0094] (8) the sense strand comprises the sequence shown in SEQ ID NO. 11, and the antisense strand comprises the sequence shown in SEQ ID NO. 109;

[0095] (9) the sense strand comprises the sequence shown in SEQ ID NO. 13, and the antisense strand comprises the sequence shown in SEQ ID NO. 111;

[0096] (10) the sense strand comprises the sequence shown in SEQ ID NO. 14, and the antisense strand comprises the sequence shown in SEQ ID NO. 112;

[0097] (11) the sense strand comprises the sequence shown in SEQ ID NO. 15, and the antisense strand comprises the sequence shown in SEQ ID NO. 113;

[0098] (12) the sense strand comprises the sequence shown in SEQ ID NO. 16, and the antisense strand comprises the sequence shown in SEQ ID NO. 114;

[0099] (13) the sense strand comprises the sequence shown in SEQ ID NO. 22, and the antisense strand comprises the sequence shown in SEQ ID NO. 120;

[0100] (14) the sense strand comprises the sequence shown in SEQ ID NO. 23, and the antisense strand comprises the sequence shown in SEQ ID NO. 121;

[0101] (15) the sense strand comprises the sequence shown in SEQ ID NO. 25, and the antisense strand comprises the sequence shown in SEQ ID NO. 123;

[0102] (16) the sense strand comprises the sequence shown in SEQ ID NO. 31, and the antisense strand comprises the sequence shown in SEQ ID NO. 129;

[0103] (17) the sense strand comprises the sequence shown in SEQ ID NO. 33, and the antisense strand comprises the sequence shown in SEQ ID NO. 131;

[0104] (18) the sense strand comprises the sequence shown in SEQ ID NO. 34, and the antisense strand comprises the sequence shown in SEQ ID NO. 132;

[0105] (19) the sense strand comprises the sequence shown in SEQ ID NO. 36, and the antisense strand comprises the sequence shown in SEQ ID NO. 134;

[0106] (20) the sense strand comprises the sequence shown in SEQ ID NO. 37, and the antisense strand comprises the sequence shown in SEQ ID NO. 135;

[0107] (21) the sense strand comprises the sequence shown in SEQ ID NO. 38, and the antisense strand comprises the sequence shown in SEQ ID NO. 136;

[0108] (22) the sense strand comprises the sequence shown in SEQ ID NO. 39, and the antisense strand comprises the sequence shown in SEQ ID NO. 137;

[0109] (23) the sense strand comprises the sequence shown in SEQ ID NO. 40, and the antisense strand comprises the sequence shown in SEQ ID NO. 138;

[0110] (24) the sense strand comprises the sequence shown in SEQ ID NO. 42, and the antisense strand comprises the sequence shown in SEQ ID NO. 140;

[0111] (25) the sense strand comprises the sequence shown in SEQ ID NO. 43, and the antisense strand comprises the sequence shown in SEQ ID NO. 141;

[0112] (26) the sense strand comprises the sequence shown in SEQ ID NO. 44, and the antisense strand comprises the sequence shown in SEQ ID NO. 142;

[0113] (27) the sense strand comprises the sequence shown in SEQ ID NO. 45, and the antisense strand comprises the sequence shown in SEQ ID NO. 143;

[0114] (28) the sense strand comprises the sequence shown in SEQ ID NO. 46, and the antisense strand comprises the sequence shown in SEQ ID NO. 144;

[0115] (29) the sense strand comprises the sequence shown in SEQ ID NO. 52, and the antisense strand comprises the sequence shown in SEQ ID NO. 150;

[0116] (30) the sense strand comprises the sequence shown in SEQ ID NO. 53, and the antisense strand comprises the sequence shown in SEQ ID NO. 151;

[0117] (31) the sense strand comprises the sequence shown in SEQ ID NO. 54, and the antisense strand comprises the sequence shown in SEQ ID NO. 152;

[0118] (32) the sense strand comprises the sequence shown in SEQ ID NO. 55, and the antisense strand comprises the sequence shown in SEQ ID NO. 153;

[0119] (33) the sense strand comprises the sequence shown in SEQ ID NO. 56, and the antisense strand comprises the sequence shown in SEQ ID NO. 154;

[0120] (34) the sense strand comprises the sequence shown in SEQ ID NO. 58, and the antisense strand comprises the sequence shown in SEQ ID NO. 156;

[0121] (35) the sense strand comprises the sequence shown in SEQ ID NO. 59, and the antisense strand comprises the sequence shown in SEQ ID NO. 157;

[0122] (36) the sense strand comprises the sequence shown in SEQ ID NO. 60, and the antisense strand comprises the sequence shown in SEQ ID NO. 158;

[0123] (37) the sense strand comprises the sequence shown in SEQ ID NO. 61, and the antisense strand comprises the sequence shown in SEQ ID NO. 159;

[0124] (38) the sense strand comprises the sequence shown in SEQ ID NO. 62, and the antisense strand comprises the sequence shown in SEQ ID NO. 160;

[0125] (39) the sense strand comprises the sequence shown in SEQ ID NO. 63, and the antisense strand comprises the sequence shown in SEQ ID NO. 161;

[0126] (40) the sense strand comprises the sequence shown in SEQ ID NO. 65, and the antisense strand comprises the sequence shown in SEQ ID NO. 163;

[0127] (41) the sense strand comprises the sequence shown in SEQ ID NO. 67, and the antisense strand comprises the sequence shown in SEQ ID NO. 165;

[0128] (42) the sense strand comprises the sequence shown in SEQ ID NO. 72, and the antisense strand comprises the sequence shown in SEQ ID NO. 170;

[0129] (43) the sense strand comprises the sequence shown in SEQ ID NO. 73, and the antisense strand comprises the sequence shown in SEQ ID NO. 171;

[0130] (44) the sense strand comprises the sequence shown in SEQ ID NO. 74, and the antisense strand comprises the sequence shown in SEQ ID NO. 172;

[0131] (45) the sense strand comprises the sequence shown in SEQ ID NO. 75, and the antisense strand comprises the sequence shown in SEQ ID NO. 173;

[0132] (46) the sense strand comprises the sequence shown in SEQ ID NO. 76, and the antisense strand comprises the sequence shown in SEQ ID NO. 174;

[0133] (47) the sense strand comprises the sequence shown in SEQ ID NO. 77, and the antisense strand comprises the sequence shown in SEQ ID NO. 175;

[0134] (48) the sense strand comprises the sequence shown in SEQ ID NO. 78, and the antisense strand comprises the sequence shown in SEQ ID NO. 176;

[0135] (49) the sense strand comprises the sequence shown in SEQ ID NO. 79, and the antisense strand comprises the sequence shown in SEQ ID NO. 177;

[0136] (50) the sense strand comprises the sequence shown in SEQ ID NO. 80, and the antisense strand comprises the sequence shown in SEQ ID NO. 178;

[0137] (51) the sense strand comprises the sequence shown in SEQ ID NO. 83, and the antisense strand comprises the sequence shown in SEQ ID NO. 181;

[0138] (52) the sense strand comprises the sequence shown in SEQ ID NO. 84, and the antisense strand comprises the sequence shown in SEQ ID NO. 182;

[0139] (53) the sense strand comprises the sequence shown in SEQ ID NO. 85, and the antisense strand comprises the sequence shown in SEQ ID NO. 183;

[0140] (54) the sense strand comprises the sequence shown in SEQ ID NO. 86, and the antisense strand comprises the sequence shown in SEQ ID NO. 184;

[0141] (55) the sense strand comprises the sequence shown in SEQ ID NO. 87, and the antisense strand comprises the sequence shown in SEQ ID NO. 185;

[0142] (56) the sense strand comprises the sequence shown in SEQ ID NO. 90, and the antisense strand comprises the sequence shown in SEQ ID NO. 188;

[0143] (57) the sense strand comprises the sequence shown in SEQ ID NO. 93, and the antisense strand comprises the sequence shown in SEQ ID NO. 191;

[0144] (58) the sense strand comprises the sequence shown in SEQ ID NO. 94, and the antisense strand comprises the sequence shown in SEQ ID NO. 192;

[0145] (59) The sense strand comprises the sequence shown in SEQ ID NO.95, and the antisense strand comprises the sequence shown in SEQ ID NO.193.

[0146] In some preferred embodiments, the oligonucleotide is selected from any one of the following sense and antisense strand combinations:

[0147] (1) the sense strand comprises the sequence shown in SEQ ID NO. 7, and the antisense strand comprises the sequence shown in SEQ ID NO. 105;

[0148] (2) the sense strand comprises the sequence shown in SEQ ID NO. 33, and the antisense strand comprises the sequence shown in SEQ ID NO. 131;

[0149] (3) the sense strand comprises the sequence shown in SEQ ID NO. 34, and the antisense strand comprises the sequence shown in SEQ ID NO. 132;

[0150] (4) the sense strand comprises the sequence shown in SEQ ID NO. 38, and the antisense strand comprises the sequence shown in SEQ ID NO. 136;

[0151] (5) the sense strand comprises the sequence shown in SEQ ID NO. 43, and the antisense strand comprises the sequence shown in SEQ ID NO. 141;

[0152] (6) the sense strand comprises the sequence shown in SEQ ID NO. 52, and the antisense strand comprises the sequence shown in SEQ ID NO. 150;

[0153] (7) the sense strand comprises the sequence shown in SEQ ID NO. 53, and the antisense strand comprises the sequence shown in SEQ ID NO. 151;

[0154] (8) the sense strand comprises the sequence shown in SEQ ID NO. 59, and the antisense strand comprises the sequence shown in SEQ ID NO. 157;

[0155] (9) the sense strand comprises the sequence shown in SEQ ID NO. 60, and the antisense strand comprises the sequence shown in SEQ ID NO. 158;

[0156] (10) the sense strand comprises the sequence shown in SEQ ID NO. 61, and the antisense strand comprises the sequence shown in SEQ ID NO. 159;

[0157] (11) the sense strand comprises the sequence shown in SEQ ID NO. 62, and the antisense strand comprises the sequence shown in SEQ ID NO. 160;

[0158] (12) the sense strand comprises the sequence shown in SEQ ID NO. 63, and the antisense strand comprises the sequence shown in SEQ ID NO. 161;

[0159] (13) the sense strand comprises the sequence shown in SEQ ID NO. 65, and the antisense strand comprises the sequence shown in SEQ ID NO. 163;

[0160] (14) the sense strand comprises the sequence shown in SEQ ID NO. 67, and the antisense strand comprises the sequence shown in SEQ ID NO. 165;

[0161] (15) the sense strand comprises the sequence shown in SEQ ID NO. 72, and the antisense strand comprises the sequence shown in SEQ ID NO. 170;

[0162] (16) the sense strand comprises the sequence shown in SEQ ID NO. 73, and the antisense strand comprises the sequence shown in SEQ ID NO. 171;

[0163] (17) the sense strand comprises the sequence shown in SEQ ID NO. 74, and the antisense strand comprises the sequence shown in SEQ ID NO. 172;

[0164] (18) the sense strand comprises the sequence shown in SEQ ID NO. 75, and the antisense strand comprises the sequence shown in SEQ ID NO. 173;

[0165] (19) the sense strand comprises the sequence shown in SEQ ID NO. 77, and the antisense strand comprises the sequence shown in SEQ ID NO. 175;

[0166] (20) the sense strand comprises the sequence shown in SEQ ID NO. 78, and the antisense strand comprises the sequence shown in SEQ ID NO. 176;

[0167] (21) the sense strand comprises the sequence shown in SEQ ID NO. 79, and the antisense strand comprises the sequence shown in SEQ ID NO. 177;

[0168] (22) the sense strand comprises the sequence shown in SEQ ID NO. 83, and the antisense strand comprises the sequence shown in SEQ ID NO. 181;

[0169] (23) the sense strand comprises the sequence shown in SEQ ID NO. 87, and the antisense strand comprises the sequence shown in SEQ ID NO. 185;

[0170] (24) the sense strand comprises the sequence shown in SEQ ID NO. 90, and the antisense strand comprises the sequence shown in SEQ ID NO. 188;

[0171] (25) The sense strand comprises the sequence shown in SEQ ID NO.94, and the antisense strand comprises the sequence shown in SEQ ID NO.192.

[0172] In some preferred embodiments, the oligonucleotide is selected from any one of the following sense and antisense strand combinations:

[0173] (1) the sense strand comprises the sequence shown in SEQ ID NO. 33, and the antisense strand comprises the sequence shown in SEQ ID NO. 131;

[0174] (2) the sense strand comprises the sequence shown in SEQ ID NO. 34, and the antisense strand comprises the sequence shown in SEQ ID NO. 132;

[0175] (3) the sense strand comprises the sequence shown in SEQ ID NO. 38, and the antisense strand comprises the sequence shown in SEQ ID NO. 136;

[0176] (4) the sense strand comprises the sequence shown in SEQ ID NO. 43, and the antisense strand comprises the sequence shown in SEQ ID NO. 141;

[0177] (5) the sense strand comprises the sequence shown in SEQ ID NO. 53, and the antisense strand comprises the sequence shown in SEQ ID NO. 151;

[0178] (6) the sense strand comprises the sequence shown in SEQ ID NO. 60, and the antisense strand comprises the sequence shown in SEQ ID NO. 158;

[0179] (7) the sense strand comprises the sequence shown in SEQ ID NO. 61, and the antisense strand comprises the sequence shown in SEQ ID NO. 159;

[0180] (8) the sense strand comprises the sequence shown in SEQ ID NO. 83, and the antisense strand comprises the sequence shown in SEQ ID NO. 181;

[0181] (9) The sense strand comprises the sequence shown in SEQ ID NO.90, and the antisense strand comprises the sequence shown in SEQ ID NO.188.

[0182] In some embodiments, the oligonucleotide comprises any one selected from the following sense and antisense strand combinations:

[0183] (1) the sense strand comprises the sequence shown in SEQ ID NO. 198, and the antisense strand comprises the sequence shown in SEQ ID NO. 261;

[0184] (2) the sense strand comprises the sequence shown in SEQ ID NO. 199, and the antisense strand comprises the sequence shown in SEQ ID NO. 262;

[0185] (3) the sense strand comprises the sequence shown in SEQ ID NO. 200, and the antisense strand comprises the sequence shown in SEQ ID NO. 263;

[0186] (4) the sense strand comprises the sequence shown in SEQ ID NO. 201, and the antisense strand comprises the sequence shown in SEQ ID NO. 264;

[0187] (5) the sense strand comprises the sequence shown in SEQ ID NO. 202, and the antisense strand comprises the sequence shown in SEQ ID NO. 265;

[0188] (6) the sense strand comprises the sequence shown in SEQ ID NO. 203, and the antisense strand comprises the sequence shown in SEQ ID NO. 266;

[0189] (7) the sense strand comprises the sequence shown in SEQ ID NO. 204, and the antisense strand comprises the sequence shown in SEQ ID NO. 267;

[0190] (8) the sense strand comprises the sequence shown in SEQ ID NO. 205, and the antisense strand comprises the sequence shown in SEQ ID NO. 268;

[0191] (9) the sense strand comprises the sequence shown in SEQ ID NO. 206, and the antisense strand comprises the sequence shown in SEQ ID NO. 269;

[0192] (10) the sense strand comprises the sequence shown in SEQ ID NO. 207, and the antisense strand comprises the sequence shown in SEQ ID NO. 270;

[0193] (11) the sense strand comprises the sequence shown in SEQ ID NO. 208, and the antisense strand comprises the sequence shown in SEQ ID NO. 271;

[0194] (12) the sense strand comprises the sequence shown in SEQ ID NO. 209, and the antisense strand comprises the sequence shown in SEQ ID NO. 272;

[0195] (13) the sense strand comprises the sequence shown in SEQ ID NO. 210, and the antisense strand comprises the sequence shown in SEQ ID NO. 273;

[0196] (14) the sense strand comprises the sequence shown in SEQ ID NO. 211, and the antisense strand comprises the sequence shown in SEQ ID NO. 274;

[0197] (15) the sense strand comprises the sequence shown in SEQ ID NO. 212, and the antisense strand comprises the sequence shown in SEQ ID NO. 275;

[0198] (16) the sense strand comprises the sequence shown in SEQ ID NO. 214, and the antisense strand comprises the sequence shown in SEQ ID NO. 277;

[0199] (17) the sense strand comprises the sequence shown in SEQ ID NO. 215, and the antisense strand comprises the sequence shown in SEQ ID NO. 278;

[0200] (18) the sense strand comprises the sequence shown in SEQ ID NO. 216, and the antisense strand comprises the sequence shown in SEQ ID NO. 279;

[0201] (19) the sense strand comprises the sequence shown in SEQ ID NO. 217, and the antisense strand comprises the sequence shown in SEQ ID NO. 280;

[0202] (20) the sense strand comprises the sequence shown in SEQ ID NO. 218, and the antisense strand comprises the sequence shown in SEQ ID NO. 281;

[0203] (21) the sense strand comprises the sequence shown in SEQ ID NO. 219, and the antisense strand comprises the sequence shown in SEQ ID NO. 282;

[0204] (22) the sense strand comprises the sequence shown in SEQ ID NO. 220, and the antisense strand comprises the sequence shown in SEQ ID NO. 283;

[0205] (23) the sense strand comprises the sequence shown in SEQ ID NO. 221, and the antisense strand comprises the sequence shown in SEQ ID NO. 284;

[0206] (24) the sense strand comprises the sequence shown in SEQ ID NO. 223, and the antisense strand comprises the sequence shown in SEQ ID NO. 286;

[0207] (25) the sense strand comprises the sequence shown in SEQ ID NO. 224, and the antisense strand comprises the sequence shown in SEQ ID NO. 287;

[0208] (26) the sense strand comprises the sequence shown in SEQ ID NO. 225, and the antisense strand comprises the sequence shown in SEQ ID NO. 288;

[0209] (27) the sense strand comprises the sequence shown in SEQ ID NO. 226, and the antisense strand comprises the sequence shown in SEQ ID NO. 289;

[0210] (28) the sense strand comprises the sequence shown in SEQ ID NO. 227, and the antisense strand comprises the sequence shown in SEQ ID NO. 290;

[0211] (29) the sense strand comprises the sequence shown in SEQ ID NO. 228, and the antisense strand comprises the sequence shown in SEQ ID NO. 291;

[0212] (30) the sense strand comprises the sequence shown in SEQ ID NO. 229, and the antisense strand comprises the sequence shown in SEQ ID NO. 292;

[0213] (31) the sense strand comprises the sequence shown in SEQ ID NO. 230, and the antisense strand comprises the sequence shown in SEQ ID NO. 293;

[0214] (32) the sense strand comprises the sequence shown in SEQ ID NO. 231, and the antisense strand comprises the sequence shown in SEQ ID NO. 294;

[0215] (33) the sense strand comprises the sequence shown in SEQ ID NO. 232, and the antisense strand comprises the sequence shown in SEQ ID NO. 295;

[0216] (34) the sense strand comprises the sequence shown in SEQ ID NO. 233, and the antisense strand comprises the sequence shown in SEQ ID NO. 296;

[0217] (35) the sense strand comprises the sequence shown in SEQ ID NO. 234, and the antisense strand comprises the sequence shown in SEQ ID NO. 297;

[0218] (36) the sense strand comprises the sequence shown in SEQ ID NO. 235, and the antisense strand comprises the sequence shown in SEQ ID NO. 298;

[0219] (37) the sense strand comprises the sequence shown in SEQ ID NO. 236, and the antisense strand comprises the sequence shown in SEQ ID NO. 299;

[0220] (38) the sense strand comprises the sequence shown in SEQ ID NO. 237, and the antisense strand comprises the sequence shown in SEQ ID NO. 300;

[0221] (39) the sense strand comprises the sequence shown in SEQ ID NO. 238, and the antisense strand comprises the sequence shown in SEQ ID NO. 301;

[0222] (40) the sense strand comprises the sequence shown in SEQ ID NO. 239, and the antisense strand comprises the sequence shown in SEQ ID NO. 302;

[0223] (41) the sense strand comprises the sequence shown in SEQ ID NO. 240, and the antisense strand comprises the sequence shown in SEQ ID NO. 303;

[0224] (42) the sense strand comprises the sequence shown in SEQ ID NO. 241, and the antisense strand comprises the sequence shown in SEQ ID NO. 304;

[0225] (43) the sense strand comprises the sequence shown in SEQ ID NO. 242, and the antisense strand comprises the sequence shown in SEQ ID NO. 305;

[0226] (44) the sense strand comprises the sequence shown in SEQ ID NO. 243, and the antisense strand comprises the sequence shown in SEQ ID NO. 306;

[0227] (45) the sense strand comprises the sequence shown in SEQ ID NO. 244, and the antisense strand comprises the sequence shown in SEQ ID NO. 307;

[0228] (46) the sense strand comprises the sequence shown in SEQ ID NO. 245, and the antisense strand comprises the sequence shown in SEQ ID NO. 308;

[0229] (47) the sense strand comprises the sequence shown in SEQ ID NO. 246, and the antisense strand comprises the sequence shown in SEQ ID NO. 309;

[0230] (48) the sense strand comprises the sequence shown in SEQ ID NO. 247, and the antisense strand comprises the sequence shown in SEQ ID NO. 310;

[0231] (49) the sense strand comprises the sequence shown in SEQ ID NO. 248, and the antisense strand comprises the sequence shown in SEQ ID NO. 311;

[0232] (50) the sense strand comprises the sequence shown in SEQ ID NO. 249, and the antisense strand comprises the sequence shown in SEQ ID NO. 312;

[0233] (51) the sense strand comprises the sequence shown in SEQ ID NO. 250, and the antisense strand comprises the sequence shown in SEQ ID NO. 313;

[0234] (52) the sense strand comprises the sequence shown in SEQ ID NO. 251, and the antisense strand comprises the sequence shown in SEQ ID NO. 314;

[0235] (53) the sense strand comprises the sequence shown in SEQ ID NO. 252, and the antisense strand comprises the sequence shown in SEQ ID NO. 315;

[0236] (54) the sense strand comprises the sequence shown in SEQ ID NO. 253, and the antisense strand comprises the sequence shown in SEQ ID NO. 316;

[0237] (55) the sense strand comprises the sequence shown in SEQ ID NO. 254, and the antisense strand comprises the sequence shown in SEQ ID NO. 317;

[0238] (56) the sense strand comprises the sequence shown in SEQ ID NO. 255, and the antisense strand comprises the sequence shown in SEQ ID NO. 318;

[0239] (57) the sense strand comprises the sequence shown in SEQ ID NO. 256, and the antisense strand comprises the sequence shown in SEQ ID NO. 319;

[0240] (58) the sense strand comprises the sequence shown in SEQ ID NO. 257, and the antisense strand comprises the sequence shown in SEQ ID NO. 320;

[0241] (59) The sense strand comprises the sequence shown in SEQ ID NO. 258, and the antisense strand comprises the sequence shown in SEQ ID NO. 321.

[0242] In some preferred embodiments, the oligonucleotide comprises any one selected from the following sense and antisense strand combinations:

[0243] (1) the sense strand comprises the sequence shown in SEQ ID NO. 202, and the antisense strand comprises the sequence shown in SEQ ID NO. 265;

[0244] (2) the sense strand comprises the sequence shown in SEQ ID NO. 215, and the antisense strand comprises the sequence shown in SEQ ID NO. 278;

[0245] (3) the sense strand comprises the sequence shown in SEQ ID NO. 216, and the antisense strand comprises the sequence shown in SEQ ID NO. 279;

[0246] (4) the sense strand comprises the sequence shown in SEQ ID NO. 219, and the antisense strand comprises the sequence shown in SEQ ID NO. 282;

[0247] (5) the sense strand comprises the sequence shown in SEQ ID NO. 224, and the antisense strand comprises the sequence shown in SEQ ID NO. 287;

[0248] (6) the sense strand comprises the sequence shown in SEQ ID NO. 228, and the antisense strand comprises the sequence shown in SEQ ID NO. 291;

[0249] (7) the sense strand comprises the sequence shown in SEQ ID NO. 229, and the antisense strand comprises the sequence shown in SEQ ID NO. 292;

[0250] (8) the sense strand comprises the sequence shown in SEQ ID NO. 234, and the antisense strand comprises the sequence shown in SEQ ID NO. 297;

[0251] (9) the sense strand comprises the sequence shown in SEQ ID NO. 235, and the antisense strand comprises the sequence shown in SEQ ID NO. 298;

[0252] (10) the sense strand comprises the sequence shown in SEQ ID NO. 236, and the antisense strand comprises the sequence shown in SEQ ID NO. 299;

[0253] (11) the sense strand comprises the sequence shown in SEQ ID NO. 237, and the antisense strand comprises the sequence shown in SEQ ID NO. 300;

[0254] (12) the sense strand comprises the sequence shown in SEQ ID NO. 238, and the antisense strand comprises the sequence shown in SEQ ID NO. 301;

[0255] (13) the sense strand comprises the sequence shown in SEQ ID NO. 239, and the antisense strand comprises the sequence shown in SEQ ID NO. 302;

[0256] (14) the sense strand comprises the sequence shown in SEQ ID NO. 240, and the antisense strand comprises the sequence shown in SEQ ID NO. 303;

[0257] (15) the sense strand comprises the sequence shown in SEQ ID NO. 241, and the antisense strand comprises the sequence shown in SEQ ID NO. 304;

[0258] (16) the sense strand comprises the sequence shown in SEQ ID NO. 242, and the antisense strand comprises the sequence shown in SEQ ID NO. 305;

[0259] (17) the sense strand comprises the sequence shown in SEQ ID NO. 243, and the antisense strand comprises the sequence shown in SEQ ID NO. 306;

[0260] (18) the sense strand comprises the sequence shown in SEQ ID NO. 244, and the antisense strand comprises the sequence shown in SEQ ID NO. 307;

[0261] (19) the sense strand comprises the sequence shown in SEQ ID NO. 246, and the antisense strand comprises the sequence shown in SEQ ID NO. 309;

[0262] (20) the sense strand comprises the sequence shown in SEQ ID NO. 247, and the antisense strand comprises the sequence shown in SEQ ID NO. 310;

[0263] (21) the sense strand comprises the sequence shown in SEQ ID NO. 248, and the antisense strand comprises the sequence shown in SEQ ID NO. 311;

[0264] (22) the sense strand comprises the sequence shown in SEQ ID NO. 250, and the antisense strand comprises the sequence shown in SEQ ID NO. 313;

[0265] (23) the sense strand comprises the sequence shown in SEQ ID NO. 254, and the antisense strand comprises the sequence shown in SEQ ID NO. 317;

[0266] (24) the sense strand comprises the sequence shown in SEQ ID NO. 255, and the antisense strand comprises the sequence shown in SEQ ID NO. 318;

[0267] (25) The positive strand comprises the sequence shown in SEQ ID NO. 257, and the antisense strand comprises the sequence shown in SEQ ID NO. 320

[0268] In some preferred embodiments, the oligonucleotide comprises any one selected from the following sense and antisense strand combinations:

[0269] (1) the sense strand comprises the sequence shown in SEQ ID NO. 215, and the antisense strand comprises the sequence shown in SEQ ID NO. 278;

[0270] (2) the sense strand comprises the sequence shown in SEQ ID NO. 216, and the antisense strand comprises the sequence shown in SEQ ID NO. 279;

[0271] (3) the sense strand comprises the sequence shown in SEQ ID NO. 219, and the antisense strand comprises the sequence shown in SEQ ID NO. 282;

[0272] (4) the sense strand comprises the sequence shown in SEQ ID NO. 224, and the antisense strand comprises the sequence shown in SEQ ID NO. 287;

[0273] (5) the sense strand comprises the sequence shown in SEQ ID NO. 229, and the antisense strand comprises the sequence shown in SEQ ID NO. 292;

[0274] (6) the sense strand comprises the sequence shown in SEQ ID NO. 235, and the antisense strand comprises the sequence shown in SEQ ID NO. 298;

[0275] (7) the sense strand comprises the sequence shown in SEQ ID NO. 236, and the antisense strand comprises the sequence shown in SEQ ID NO. 299;

[0276] (8) the sense strand comprises the sequence shown in SEQ ID NO. 250, and the antisense strand comprises the sequence shown in SEQ ID NO. 313;

[0277] (9) The sense strand comprises the sequence shown in SEQ ID NO. 255, and the antisense strand comprises the sequence shown in SEQ ID NO. 318.

[0278] On the other hand, the present disclosure provides a conjugate or a pharmaceutically acceptable salt thereof for reducing HSD17B13 gene expression, which comprises: (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 above-mentioned siRNA and a conjugated group connected to the siRNA. The term "oligonucleotide salt" refers to an oligonucleotide compound in the form of a salt. Oligonucleotide salts include salts of oligonucleotide conjugated compounds and salts of unconjugated oligonucleotide compounds. Oligonucleotide salts are advantageously present in the form of solid powder.

[0279] 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.

[0280] 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.

[0281] 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.

[0282] In some embodiments, at least one nucleotide of the oligonucleotide is conjugated to one or more targeting ligands.

[0283] In some embodiments, the targeting ligand comprises a carbohydrate, an amino sugar, cholesterol, a polypeptide, or a lipid.

[0284] In some embodiments, the targeting ligand comprises an N-acetylgalactosamine (GalNAc) moiety.

[0285] In some embodiments, the GalNac moiety is a monovalent GalNAc moiety, a divalent GalNAc moiety, a trivalent GalNAc moiety, or a tetravalent GalNAc moiety.

[0286] In some embodiments, the targeting ligand is selected from L96 and (NAG37)s. 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.

[0287] In another aspect, the present disclosure provides a composition comprising the aforementioned oligonucleotide or a pharmaceutically acceptable salt thereof or the aforementioned conjugate, and optionally a pharmaceutically acceptable carrier.

[0288] In some embodiments, the composition is administered orally, intravenously, subcutaneously, or intramuscularly.

[0289] In some embodiments, the HSD17B13-related disease is selected from non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis and cirrhosis, and complications associated with these diseases such as type 2 diabetes, cardiovascular disease, chronic kidney disease, etc.

[0290] In another aspect, the present disclosure provides use of the aforementioned oligonucleotide or a pharmaceutically acceptable salt or composition thereof in the preparation of a medicament for treating and / or preventing HSD17B13-related diseases.

[0291] In some embodiments, the HSD17B13-related disease is selected from non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, and cirrhosis.

[0292] In some embodiments, administration of an oligonucleotide as described herein results in a reduction in the level of HSD17B13 expression in a cell. In some embodiments, the reduction in the level of HSD17B13 expression can be a reduction 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 HSD17B13. An appropriate control level can be the level of HSD17B13 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, HSD17B13 mRNA levels in hepatocytes 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 introduction of the oligonucleotide into the cells.

[0293] 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., HSD17B13), e.g., confirming the inhibitory effect on the human gene by nadir HSD17B13 expression following subcutaneous injection when administered as a single dose, e.g., at 2 mg / kg. Such a system is useful when the nucleic acid sequences of the human gene and the model animal gene are sufficiently close that the human iRNA provides effective knockdown of the model animal gene.

[0294] Inhibition of HSD17B13 gene expression can be represented 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 HSD17B13 gene is transcribed and which is treated (e.g., by contacting one or more cells with an iRNA of the disclosure, or by administering an iRNA of the disclosure to a subject in which cells are or were present) such that expression of the HSD17B13 gene is inhibited compared to a cell line that is substantially identical to the cell line but is not treated (control cells that are not treated with the iRNA or that are not treated with an iRNA targeting the gene of interest). In a preferred embodiment, inhibition is assessed using siRNA concentrations of 0.5 nM and 0.05 nM in species-matched cell lines as described in Example 2, and is represented by 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: ΔCT = CT HSD17B13 -CT GAPDH△△CT=△CT 处理细胞 -△CT 对照细胞

[0295] mRNA level = 2^-△△CT

[0296] In other embodiments, inhibition of HSD17B13 gene expression can be assessed based on a reduction in a parameter functionally associated with HSD17B13 gene expression, e.g., HSD17B13 protein levels in blood or serum from a subject. HSD17B13 gene silencing can be determined in any cell expressing HSD17B13, whether endogenous or heterologous from an expression construct, and by any assay known in the art.

[0297] Inhibition of HSD17B13 protein expression can be demonstrated by a decrease in the level of HSD17B13 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 protein levels in control cells or cell populations, or as a change in protein levels in a sample from a subject (e.g., blood or serum from a subject). Percent inhibition of mRNA = (protein expression level) 处理细胞 -Protein expression 对照细胞 ) / protein expression 对照细胞 *100%

[0298] 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 therapeutically benefit from, for example, a reduction in the amount of HSD17B13 protein.

[0299] 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.

[0300] 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.

[0301] 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.

[0302] In some embodiments, the oligonucleotide is administered at a fixed dose of about 10 mg to about 800 mg. In some embodiments, the oligonucleotide is administered to a subject at a fixed dose of about 10 to 50 mg, 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 10 mg, about 50 mg, about 100 mg, about 200 mg, about 300 mg, about 400 mg, 500 mg, about 600 mg, about 700 mg, or about 800 mg.

[0303] 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.

[0304] 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.

[0305] In another aspect, the present disclosure provides a method for inhibiting HSD17B13 gene expression in a subject, comprising administering to the subject the aforementioned oligonucleotide or a pharmaceutically acceptable salt thereof, or the aforementioned conjugate or a pharmaceutically acceptable salt thereof, or the aforementioned composition.

[0306] In another aspect, the present disclosure provides a method for preventing or treating a subject suffering from an HSD17B13-related disease, comprising administering to the subject the aforementioned oligonucleotide or a pharmaceutically acceptable salt thereof, the aforementioned conjugate or a pharmaceutically acceptable salt thereof, or the aforementioned composition.

[0307] In another aspect, the present disclosure provides a method for inhibiting HSD17B13 gene expression in a subject, comprising administering to the subject a fixed dose of about 10 mg to about 800 mg of an oligonucleotide or a salt thereof, or a conjugate or a salt thereof, or a composition comprising the same.

[0308] In another aspect, the present disclosure provides a method for treating a subject suffering from an HSD17B13-related disorder, the method comprising administering to the subject a fixed dose of about 10 mg to about 800 mg of an oligonucleotide or a salt thereof, or a conjugate thereof or a salt thereof, or a composition comprising the same. In some embodiments, the HSD17B13-related disease is selected from non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis and cirrhosis, and complications associated with these diseases such as type 2 diabetes, cardiovascular disease, chronic kidney disease, etc.

[0309] In some embodiments, the method further comprises co-administering to the subject an additional therapeutic agent for treating an HSD17B13-related disease.

[0310] In some embodiments, the method further comprises co-administering to the subject an additional non-HSD17B13 siRNA therapeutic agent, wherein the non-HSD17B13 siRNA therapeutic agent comprises one or more of the following, such as glitazones, vitamin E, liraglutide, metformin, statins, pentoxifylline, and obeticholic acid.

[0311] 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.

[0312] 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.

[0313] Example 1

[0314] Unless the source of a reagent is specifically given herein, such reagent can be obtained from any molecular biology reagent supplier at quality / purity standards appropriate for molecular biology.

[0315] Abbreviations for nucleotide monomers used in nucleic acid sequence representations. It will be understood that these monomers, when present in an oligonucleotide, are interconnected by 5'-3' phosphodiester bonds unless otherwise indicated.

[0316] Table A. Nucleotide monomer abbreviations used in nucleic acid sequence representation

[0317] Preparation of targeting ligands

[0318] (i) L96 was prepared according to the method described in patent CN104717982B.

[0319] (ii) (NAG37)s was prepared according to the method described in patent CN113164509A.

[0320] Preparation of oligonucleotides

[0321] (1) Preparation of siRNA

[0322] 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.

[0323] Solid-phase synthesis (Figure 2): 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 Berleke Precision Instrument Co., Ltd.). Solid-phase synthesis begins at the 3' end of the sequence and sequentially couples monomers 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.

[0324] (2) Preparation of double-stranded RNA agents

[0325] (a) Synthesis of the positive chain

[0326] 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.

[0327] (b) Synthesis of antisense strand

[0328] 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.

[0329] (c) Preparation of double-stranded siRNA

[0330] 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.

[0331] According to the same method, the double-stranded siRNA agents shown in Tables 2, 3 and 4 below were prepared.

[0332] First, a computer-based algorithm was used to generate candidate oligonucleotide sequences complementary to human HSD17B13 mRNA (NM_178135.4, Table 1). Some of these sequences were also complementary to or had no more than two mismatches with cynomolgus macaque HSD17B13 mRNA (XM_005555367.3, 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; in some cases, the overhangs were non-complementary UU. 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. The first base at the 5' end of the antisense strand (the last base at the 3' end of the sense strand) of some complementary paired sequences was replaced by a base that did not match HSD17B13 mRNA.

[0333] Table 1: Human and cynomolgus monkey HSD17B13 mRNA sequences

[0334] 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 stands for 2'-deoxyadenosine 3'-phosphate; dG stands for 2'-deoxyguanosine 3'-phosphate; dC stands for 2'-deoxycytidine 3'-phosphate; and dU stands for 2'-deoxyuridine 3'-phosphate.

[0335] For nucleotide modifications: m represents 2'-methoxy; f represents 2'-deoxy-2'-fluoro; and s represents phosphorothioate.

[0336] Certain oligonucleotides may include a diol nucleic acid (GNA) modification, wherein the ribonucleotide has a diol nucleic acid structure as shown in formula (I). 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).

[0337] Certain oligonucleotides may contain 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). 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).

[0338] Some oligonucleotides have 5'-phosphate analog modified nucleotides at the 5' end. In some oligonucleotides, the 5'-phosphate analog modified nucleotides have vinyl phosphonate modified nucleotides shown in formula (XI). In some oligonucleotides, the 5'-phosphate analog modified nucleotides have vinyl phosphonate modified nucleotides shown in formula (XII). In some oligonucleotides, the 5' end has APU, which is a 5'-phosphate analog modified uridylic acid (2'-acetamido-5'-vinyl phosphonate-uridylic acid) shown in formula (XIII). In some oligonucleotides, the 5' end has VPUm, which is a 5'-phosphate analog modified uridylic acid (2'-methoxy-5'-vinyl phosphonate-uridylic acid) shown in formula (XIV).

[0339] Certain oligonucleotides may comprise inverted ribonucleotides comprising an inverted abasic residue ((invAb)) selected from the group consisting of abasic residues represented by formula (XV), (XVI), or (XVII):

[0340] Certain oligonucleotides can be conjugated to targeting ligands. The targeting ligands are selected from L96 and (NAG37)s, wherein L96 is N-[tris(GalNAc-alkyl)amidodecanoyl]-4-hydroxyprolinol (Hyp-(GalNAc-alkyl)3); and (NAG37)s is a GalNAc carrier. The structural formulas of L96 and (NAG37)s are as follows:

[0341] Table 2: Naked oligonucleotide sequences

[0342] Table 3: Modified oligonucleotides

[0343] Table 4: siRNA sequences with vectors

[0344] Example 2. In vitro activity screening of naked HSD17B13-siRNA sequences in HEK293A (psiCHECK2 dual luciferase reporter system)

[0345] (1) Cell culture and transfection:

[0346] HEK293A cells (iCell-h086, Shanghai Biotechnology Co., Ltd.) were placed in a 37°C, 5% CO2 incubator and cultured in Dulbecco's Modified Eagle Medium (iCell-h086-001b, Shanghai Biotechnology Co., Ltd.) supplemented with 10% FBS (GIBCO, 12483020) and 1% penicillin-streptomycin (GIBCO, 15140-122). Cells were digested with trypsin-EDTA (Thermo, 25200-072) until the confluence reached 90%. The cells were counted using a Countstar IC1000 counter and 190 μl of cell suspension was plated per well in a 96-well plate (Shanghai Biotechnology Co., Ltd., FCP968). The cell number was 1 × 10 4 Cells were transfected at 4 μg / well and allowed to adhere the next day. Transfection was performed using Lipofectamine 3000 Transfection Reagent (thermofisher, L3000150). Specifically, siRNA was diluted with Opti-MEM (thermofisher, 1105821) to a concentration of 0.1 μM / L or 0.01 μM / L, and psiCHECK2-hHSD17B13 plasmid (General Biotechnology (Anhui) Co., Ltd.) was diluted with Opti-MEM to 2.5 ng / μl. For system ①, 1 μl of siRNA was added to 4 μl of Opti-MEM containing the plasmid. For system ②, 0.3 μl of Lipo3000 was diluted with 5 μl of Opti-MEM. After each system was allowed to stand for 5 minutes, systems ① and ② were mixed, allowed to stand for 15 minutes, and then added dropwise to a 96-well plate. The final siRNA concentration was 5 nM or 0.5 nM, and the plasmid concentration was 10 ng per well. The 96-well plate was placed in an incubator and incubated for 24 h.

[0347] (2) Detection

[0348] 24 h after transfection, the original culture medium in the 96-well plate was discarded, each well was rinsed with 200 μL PBS (gibco, 20012027) and discarded, and 50 μL 1× Mix Luciferase Assay Reagent (Progega, E2940) with 50 μL PBS, add to the cell wells, and lyse on a shaker for at least 15 minutes. Read the firefly fluorescence signal in a microplate reader (BioTek, Synerg H1M). Add 50 μL stop solution to each well. Stop& Reagent (Progega, E2940) and read the Renilla fluorescence signal.

[0349] (3) Data statistical analysis: Calculation of relative luciferase ratio (Ratio): Relative luciferase ratio = Renilla luciferase activity (RL) / Firefly luciferase activity (FL) Inhibition efficiency (%) = (1-(Ratio 给药组 / Ratio 对照组 ))*100%.

[0350] The cell screening results are shown in Table 5.

[0351] As shown in Table 5, when the dosage was 0.5 nM, except for AL0201005, AL0201017, AL0201018, AL0201020, AL0201047, AL0201048, AL0201057, AL0201066, AL0201070, AL0201073, AL0201074, AL0201075, AL0201076, AL0201077, AL0201079, AL0201080, AL0201082, AL0201090 and AL0201097, the inhibition effect of most siRNAs was ≥80%. At a low concentration of 0.05 nM, the inhibition rate of HSD17B13 mRNA decreased, but most sequences had no significant effect on HSD17B13 mRNA. The mRNA inhibition rate can reach 70%.

[0352] Table 5: Knockdown levels of HSD17B13 naked siRNA sequences in the psiCHEC2 dual-luciferase reporter gene system

[0353] Example 3. In vitro activity screening of chemically modified HSD17B13-siRNA

[0354] The psiCHECK2 dual luciferase reporter system screening was the same as in Example 2.

[0355] (1) Cynomolgus monkey primary liver cell culture and transfection:

[0356] Before adding cells, remove the culture medium (Beijing Red Biotech Co., Ltd., HEPO24) and preheat it in a biosafety cabinet. Take 36 ml of culture medium and add 4 ml of FBS to prepare complete culture medium. Heat it in a 37°C water bath for 10 minutes. Treat with coating medium (Beijing Red Biotech Co., Ltd., HEPO44) in a CO2 incubator at 37°C for 0.5 hours. Take out the cynomolgus monkey primary hepatocytes (Beijing Red Biotech Co., Ltd., cmTCSC) from liquid nitrogen and revive the cells in a 37°C water bath. After about 2 minutes, take out the cells and transfer the cell suspension to 40 ml of preheated complete culture medium. Wash the cell cryopreservation tube with 2 ml of complete culture medium. Centrifuge the cell suspension at 180 × g for 1 minute, discard the supernatant, add 2 ml of preheated CM seeding medium (Beijing Red Biotech Co., Ltd., CMHEP054), gently blow to mix the cell suspension, and take 20 μl of cell suspension for counting. According to the counting results, the cells were diluted to 3×10 5 / ml, according to 3×10 5 Inoculate 12-well plates in 12-well plates and culture in a 37°C, 5% CO2 incubator. After 4-5 hours of adherence, aspirate the CM seeding medium and replace it with pre-warmed culture medium (Beijing Red Biotech Co., Ltd., CMHEP064) at 0.9 ml / well. Transfect 6 hours after adherence. Use Lipofectamine TM Transfection was performed using 3000 Transfection Reagent (thermofisher, L3000150). System ① was diluted with 50 μl Opti-MEM (thermofisher, 1105821) to a concentration of 100 nM or 10 nM siRNA, and system ② was diluted with 50 μl Opti-MEM to a concentration of 3 μl Lipo3000. After standing for 5 min, systems ① and ② were mixed at a siRNA concentration of 50 nM or 5 nM. After standing for 15 min, the cells were added dropwise to a 12-well plate at a final siRNA concentration of 5 nM or 0.5 nM. DMEM / F12 complete medium was replaced 4 h after transfection, and the 12-well plates were incubated in an incubator for 24 h.

[0357] (2) RNA extraction and detection

[0358] 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).

[0359] cDNA was reverse transcribed using HiScript IIQ RT SuperMix for qPCR (with gDNA wiper) (Vazyme, R223-01). A mixture of 4× gDNA wiper mix (4 μl) and 12 μl of RNA sample was prepared in an RNase-free centrifuge tube. The mixture was then filled to 16 μl with RNase-free ddH2O. Each sample was replicated at 42°C for 2 min. Four μl of 5× 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.

[0360] A mixture of 10 μl 2×ChamQ 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: pre-denaturation at 95°C for 30 seconds; amplification at 95°C for 5 seconds; 60°C for 20 seconds; 40 cycles; melting curve at 95°C for 15 seconds, 60°C for 60 seconds, and 95°C for 15 seconds.

[0361] (3) Data statistical analysis:

[0362] Export the data to EXCEL format and use CT HSD17B13 -CT GAPDH The control group was normalized, and the data were analyzed using the ΔΔCT method to calculate the fold change in relative silencing efficiency. The results are shown in Tables 6, 7, and 8. Tables 6-8 show the results of experiments conducted at different times using different batches of monkey primary cells.

[0363] When the dosage was 5 nM and 0.5 nM, AL0205005, AL0205006, AL0205007, AL0205009, AL0205010, AL0205013, AL0205018, AL0205019, AL0205020, AL0205022, AL0205023, AL0205024, AL0205027, AL0205029, AL0205031, AL0205032, AL0205034, AL0205035, AL0205037, AL0205038, AL0205039, AL0205040 The knockdown rates of 5040, AL0205041, AL0205042, AL0205043, AL0205044, AL0205045, AL0205046, AL0205047, AL0205049, AL0205050, AL0205051, AL0205052, AL0205053, AL0205054, AL0205055, AL0205056, AL0205057, AL0205058, AL0205060 and AL0205061 on HSD17B13 mRNA were all ≥70%, and the knockdown rates of some sequences on HSD17B13 mRNA could even reach 80% or even 90%.

[0364] The knockdown effect of HSD17B13 mRNA was further tested in cynomolgus monkeys. The results showed that at a very low concentration of 0.1 nM, AL0205018, AL0205019, AL0205022, AL0205027, AL0205032, AL0205038, AL0205039, AL0205053 and AL0205058 could still knock down HSD17B13 mRNA by >50%, indicating a significant inhibitory effect.

[0365] Table 6: Knockdown of HSD17B13-siRNA modified sequences in cynomolgus monkey primary hepatocytes

[0366] Table 7: Knockdown of HSD17B13-siRNA modified sequences in cynomolgus monkey primary hepatocytes

[0367] Table 8: Knockdown of HSD17B13-siRNA modified sequences in cynomolgus monkey primary hepatocytes

Claims

1. An oligonucleotide or a pharmaceutically acceptable salt thereof for reducing the expression of the HSD17B13 gene, the oligonucleotide comprising a sense strand and an antisense strand, the sense strand having a sequence with at least 80% sequence identity to the sequence shown in any one of SEQ ID NOs. 1-26, 28-40 and 42-97, or a fragment thereof, or a modified sequence of the sequence or its fragment, preferably having a sequence identity of 85%, 90%, 95%, 96%, 97%, 98%, 99% or more; the antisense strand having a sequence with at least 80% sequence identity to the sequence shown in any one of SEQ ID NOs. 99-124, 126-138 and 140-195, or a fragment thereof, or a modified sequence of the sequence or its fragment, preferably having a sequence identity of 85%, 90%, 95%, 96%, 97%, 98%, 99% or more.

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 carboxylates, alkali metal salts, ammonium salts, alkaline earth metal salts, salts formed with organic bases, and other pharmaceutically acceptable salts; Preferably, the salt is an alkali metal salt, more preferably sodium salt or potassium salt; Preferably, the salt is an alkaline earth metal salt, more preferably magnesium salt or calcium salt; Preferably, the salt is an ammonium salt, more preferably triethylamine salt.

3. The oligonucleotide or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein, The oligonucleotide contains at least one modified nucleotide; Preferably, the oligonucleotide contains 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, 2'-deoxynucleotides; Preferably, the 2'-modification is a modification selected from the following: 2'-methoxy, 2'-acetamido, 2'-aminoethyl, 2'-fluoro, 2'-O-methoxyethyl; Preferably, the oligonucleotide comprises glycol nucleic acid (GNA) modification, and the ribonucleotide has a glycol nucleic acid structure shown in formula (I), preferably the S-isomer; preferably, the oligonucleotide comprises Tgn, which is thymidine-glycol nucleic acid shown in formula (II), preferably its S-isomer; preferably, the oligonucleotide comprises Cgn, which is cytidine-glycol nucleic acid shown in formula (III), preferably its S-isomer; preferably, the oligonucleotide comprises Agn, which is adenosine-glycol nucleic acid shown in formula (IV), preferably its S-isomer; preferably, the oligonucleotide comprises Ggn, which is guanosine-glycol nucleic acid shown in formula (V), preferably its S-isomer; Preferably, the oligonucleotide contains the nucleoside-2'-phosphate shown in formula (VI); preferably, the oligonucleotide contains U-2'5', which is uridine-2'-phosphate shown in formula (VII); preferably, the oligonucleotide contains C-2'5', which is cytidine-2'-phosphate shown in formula (VIII); preferably, the oligonucleotide contains A-2'5', which is adenosine-2'-phosphate shown in formula (IX); preferably, the oligonucleotide contains G-2'5', which is guanosine-2'-phosphate shown in formula (X); Nucleoside-2'-phosphate Preferably, the oligonucleotide has a nucleotide modified with a 5'-phosphate analog at the 5'-end; preferably, the nucleotide modified with a 5'-phosphate analog has a nucleotide modified with vinylphosphonate as shown in formula (XI); preferably, the nucleotide modified with a 5'-phosphate analog has a nucleotide modified with vinylphosphonate as shown in formula (XII); preferably, the oligonucleotide has APU at the 5'-end, which is a uridylate modified with a 5'-phosphate analog as shown in formula (XIII) (2'-acetamido-5'-vinylphosphonate-uridylate); preferably, the oligonucleotide has VPU at the 5'-end, which is a uridylate modified with a 5'-phosphate analog as shown in formula (XIV) (2'-methoxy-5'-vinylphosphonate-uridylate); Preferably, the oligonucleotide comprises a 6-(3-(2-carboxyethyl)phenyl)purine-modified nucleotide; preferably, the oligonucleotide comprises Formula M, which is a 2'-O-methyl-6-(3-(2-carboxyethyl)phenyl)-purine nucleotide represented by Formula (XVII); 4. The oligonucleotide or a pharmaceutically acceptable salt thereof according to any one of claims 1-3, wherein, The oligonucleotide contains at least one modified internucleotide bond; Preferably, the 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 1-4, wherein, The sense strand comprises a sequence shown in any one selected from SEQ ID NO.1-2, 4, 6-8, 10-11, 13-16, 22-23, 25, 31, 33-34, 36-40, 42-46, 52-56, 58-63, 65, 67, 72-80, 83-87, 90, 93-95 or a modified sequence thereof; the antisense strand comprises a sequence shown in any one selected from SEQ ID NO.99, 100, 102, 104, 105, 106, 108, 109, 111, 112, 113, 114, 120, 121, 123, 129, 131, 132, 134, 135, 136, 137, 138, 140, 141, 142, 143, 144, 150, 151, 152, 153, 154, 156, 157, 158, 159, 160, 161, 163, 165, 170, 171, 172, 173, 174, 175, 176, 177, 178, 181, 182, 183, 184, 185, 188, 191, 192, 193 or a modified sequence thereof; Preferably, the sense strand comprises a sequence shown in any one selected from SEQ ID NO.7, 33, 34, 38, 43, 52, 53, 59, 60, 61, 62, 63, 65, 67, 72, 73, 74, 75, 77, 78, 79, 83, 87, 90 and 94 or a modified sequence thereof; the antisense strand comprises a sequence shown in any one selected from SEQ ID NO.105, 131, 132, 136, 141, 150, 151, 157, 158, 159, 160, 161, 163, 165, 170, 171, 172, 173, 175, 176, 177, 181, 185, 188 and 192 or a modified sequence thereof; Preferably, the modified sequence of the sense strand comprises a sequence described in any one selected from SEQ ID NO.198-212, 214-221, 223-258; the modified sequence of the antisense strand comprises a sequence described in any one selected from SEQ ID NO.261-275, 277-284, 286-321.

6. The oligonucleotide or a pharmaceutically acceptable salt thereof according to any one of claims 1-5, wherein, The oligonucleotide is selected from any one of the following combinations of sense strand and antisense strand: (1) The sense strand comprises the sequence shown in SEQ ID NO.1, and the antisense strand comprises the sequence shown in SEQ ID NO.99; (2) The sense strand comprises the sequence shown in SEQ ID NO.2, and the antisense strand comprises the sequence shown in SEQ ID NO.100; (3) The sense strand comprises the sequence shown in SEQ ID NO.4, and the antisense strand comprises the sequence shown in SEQ ID NO.102; (4) The sense strand comprises the sequence shown in SEQ ID NO.6, and the antisense strand comprises the sequence shown in SEQ ID NO.104; (5) The sense strand contains the sequence shown in SEQ ID NO.7, and the antisense strand contains the sequence shown in SEQ ID NO.105; (6) The sense strand contains the sequence shown in SEQ ID NO.8, and the antisense strand contains the sequence shown in SEQ ID NO.106; (7) The sense strand contains the sequence shown in SEQ ID NO.10, and the antisense strand contains the sequence shown in SEQ ID NO.108; (8) The sense strand contains the sequence shown in SEQ ID NO.11, and the antisense strand contains the sequence shown in SEQ ID NO.109; (9) The sense strand contains the sequence shown in SEQ ID NO.13, and the antisense strand contains the sequence shown in SEQ ID NO.111; (10) The sense strand contains the sequence shown in SEQ ID NO.14, and the antisense strand contains the sequence shown in SEQ ID NO.112; (11) The sense strand contains the sequence shown in SEQ ID NO.15, and the antisense strand contains the sequence shown in SEQ ID NO.113; (12) The sense strand contains the sequence shown in SEQ ID NO.16, and the antisense strand contains the sequence shown in SEQ ID NO.114; (13) The sense strand contains the sequence shown in SEQ ID NO.22, and the antisense strand contains the sequence shown in SEQ ID NO.120; (14) The sense strand contains the sequence shown in SEQ ID NO.23, and the antisense strand contains the sequence shown in SEQ ID NO.121; (15) The sense strand contains the sequence shown in SEQ ID NO.25, and the antisense strand contains the sequence shown in SEQ ID NO.123; (16) The sense strand contains the sequence shown in SEQ ID NO.31, and the antisense strand contains the sequence shown in SEQ ID NO.129; (17) The sense strand contains the sequence shown in SEQ ID NO.33, and the antisense strand contains the sequence shown in SEQ ID NO.131; (18) The sense strand contains the sequence shown in SEQ ID NO.34, and the antisense strand contains the sequence shown in SEQ ID NO.132; (19) The sense strand contains the sequence shown in SEQ ID NO.36, and the antisense strand contains the sequence shown in SEQ ID NO.134; (20) The sense strand contains the sequence shown in SEQ ID NO.37, and the antisense strand contains the sequence shown in SEQ ID NO.135; (21) The sense strand contains the sequence shown in SEQ ID NO.38, and the antisense strand contains the sequence shown in SEQ ID NO.136; (22) The sense strand contains the sequence shown in SEQ ID NO. 39, and the antisense strand contains the sequence shown in SEQ ID NO. 137; (23) The sense strand contains the sequence shown in SEQ ID NO. 40, and the antisense strand contains the sequence shown in SEQ ID NO. 138; (24) The sense strand contains the sequence shown in SEQ ID NO. 42, and the antisense strand contains the sequence shown in SEQ ID NO. 140; (25) The sense strand contains the sequence shown in SEQ ID NO. 43, and the antisense strand contains the sequence shown in SEQ ID NO. 141; (26) The sense strand contains the sequence shown in SEQ ID NO. 44, and the antisense strand contains the sequence shown in SEQ ID NO. 142; (27) The sense strand contains the sequence shown in SEQ ID NO. 45, and the antisense strand contains the sequence shown in SEQ ID NO. 143; (28) The sense strand contains the sequence shown in SEQ ID NO. 46, and the antisense strand contains the sequence shown in SEQ ID NO. 144; (29) The sense strand contains the sequence shown in SEQ ID NO. 52, and the antisense strand contains the sequence shown in SEQ ID NO. 150; (30) The sense strand contains the sequence shown in SEQ ID NO. 53, and the antisense strand contains the sequence shown in SEQ ID NO. 151; (31) The sense strand contains the sequence shown in SEQ ID NO. 54, and the antisense strand contains the sequence shown in SEQ ID NO. 152; (32) The sense strand contains the sequence shown in SEQ ID NO. 55, and the antisense strand contains the sequence shown in SEQ ID NO. 153; (33) The sense strand contains the sequence shown in SEQ ID NO. 56, and the antisense strand contains the sequence shown in SEQ ID NO. 154; (34) The sense strand contains the sequence shown in SEQ ID NO. 58, and the antisense strand contains the sequence shown in SEQ ID NO. 156; (35) The sense strand contains the sequence shown in SEQ ID NO. 59, and the antisense strand contains the sequence shown in SEQ ID NO. 157; (36) The sense strand contains the sequence shown in SEQ ID NO. 60, and the antisense strand contains the sequence shown in SEQ ID NO. 158; (37) The sense strand contains the sequence shown in SEQ ID NO. 61, and the antisense strand contains the sequence shown in SEQ ID NO. 159; (38) The sense strand contains the sequence shown in SEQ ID NO. 62, and the antisense strand contains the sequence shown in SEQ ID NO. 160; (39) The sense strand contains the sequence shown in SEQ ID NO.63, and the antisense strand contains the sequence shown in SEQ ID NO.161; (40) The sense strand contains the sequence shown in SEQ ID NO.65, and the antisense strand contains the sequence shown in SEQ ID NO.163; (41) The sense strand contains the sequence shown in SEQ ID NO.67, and the antisense strand contains the sequence shown in SEQ ID NO.165; (42) The sense strand contains the sequence shown in SEQ ID NO.72, and the antisense strand contains the sequence shown in SEQ ID NO.170; (43) The sense strand contains the sequence shown in SEQ ID NO.73, and the antisense strand contains the sequence shown in SEQ ID NO.171; (44) The sense strand contains the sequence shown in SEQ ID NO.74, and the antisense strand contains the sequence shown in SEQ ID NO.172; (45) The sense strand contains the sequence shown in SEQ ID NO.75, and the antisense strand contains the sequence shown in SEQ ID NO.173; (46) The sense strand contains the sequence shown in SEQ ID NO.76, and the antisense strand contains the sequence shown in SEQ ID NO.174; (47) The sense strand contains the sequence shown in SEQ ID NO.77, and the antisense strand contains the sequence shown in SEQ ID NO.175; (48) The sense strand contains the sequence shown in SEQ ID NO.78, and the antisense strand contains the sequence shown in SEQ ID NO.176; (49) The sense strand contains the sequence shown in SEQ ID NO.79, and the antisense strand contains the sequence shown in SEQ ID NO.177; (50) The sense strand contains the sequence shown in SEQ ID NO.80, and the antisense strand contains the sequence shown in SEQ ID NO.178; (51) The sense strand contains the sequence shown in SEQ ID NO.83, and the antisense strand contains the sequence shown in SEQ ID NO.181; (52) The sense strand contains the sequence shown in SEQ ID NO.84, and the antisense strand contains the sequence shown in SEQ ID NO.182; (53) The sense strand contains the sequence shown in SEQ ID NO.85, and the antisense strand contains the sequence shown in SEQ ID NO.183; (54) The sense strand contains the sequence shown in SEQ ID NO.86, and the antisense strand contains the sequence shown in SEQ ID NO.184; (55) The sense strand contains the sequence shown in SEQ ID NO.87, and the antisense strand contains the sequence shown in SEQ ID NO.185; (56) The sense strand contains the sequence shown in SEQ ID NO.90, and the antisense strand contains the sequence shown in SEQ ID NO.188; (57) The sense strand contains the sequence shown in SEQ ID NO.93, and the antisense strand contains the sequence shown in SEQ ID NO.191; (58) The sense strand contains the sequence shown in SEQ ID NO.94, and the antisense strand contains the sequence shown in SEQ ID NO.192; (59) The sense strand contains the sequence shown in SEQ ID NO.95, and the antisense strand contains the sequence shown in SEQ ID NO.193; Preferably, the oligonucleotide is selected from any of the following combinations of sense and antisense strands: (1) The sense strand contains the sequence shown in SEQ ID NO.7, and the antisense strand contains the sequence shown in SEQ ID NO.105; (2) The sense strand contains the sequence shown in SEQ ID NO.33, and the antisense strand contains the sequence shown in SEQ ID NO.131; (3) The sense strand contains the sequence shown in SEQ ID NO.34, and the antisense strand contains the sequence shown in SEQ ID NO.132; (4) The sense strand contains the sequence shown in SEQ ID NO.38, and the antisense strand contains the sequence shown in SEQ ID NO.136; (5) The sense strand contains the sequence shown in SEQ ID NO.43, and the antisense strand contains the sequence shown in SEQ ID NO.141; (6) The sense strand contains the sequence shown in SEQ ID NO.52, and the antisense strand contains the sequence shown in SEQ ID NO.150; (7) The sense strand contains the sequence shown in SEQ ID NO.53, and the antisense strand contains the sequence shown in SEQ ID NO.151; (8) The sense strand contains the sequence shown in SEQ ID NO.59, and the antisense strand contains the sequence shown in SEQ ID NO.157; (9) The sense strand contains the sequence shown in SEQ ID NO.60, and the antisense strand contains the sequence shown in SEQ ID NO.158; (10) The sense strand contains the sequence shown in SEQ ID NO.61, and the antisense strand contains the sequence shown in SEQ ID NO.159; (11) The sense strand contains the sequence shown in SEQ ID NO.62, and the antisense strand contains the sequence shown in SEQ ID NO.160; (12) The sense strand contains the sequence shown in SEQ ID NO.63, and the antisense strand contains the sequence shown in SEQ ID NO.161; (13) The sense strand contains the sequence shown in SEQ ID NO.65, and the antisense strand contains the sequence shown in SEQ ID NO.163; (14) The sense strand contains the sequence shown in SEQ ID NO.67, and the antisense strand contains the sequence shown in SEQ ID NO.165; (15) The sense strand contains the sequence shown in SEQ ID NO.72, and the antisense strand contains the sequence shown in SEQ ID NO.170; (16) The sense strand contains the sequence shown in SEQ ID NO.73, and the antisense strand contains the sequence shown in SEQ ID NO.171; (17) The sense strand contains the sequence shown in SEQ ID NO.74, and the antisense strand contains the sequence shown in SEQ ID NO.172; (18) The sense strand contains the sequence shown in SEQ ID NO.75, and the antisense strand contains the sequence shown in SEQ ID NO.173; (19) The sense strand contains the sequence shown in SEQ ID NO.77, and the antisense strand contains the sequence shown in SEQ ID NO.175; (20) The sense strand contains the sequence shown in SEQ ID NO.78, and the antisense strand contains the sequence shown in SEQ ID NO.176; (21) The sense strand contains the sequence shown in SEQ ID NO.79, and the antisense strand contains the sequence shown in SEQ ID NO.177; (22) The sense strand contains the sequence shown in SEQ ID NO.83, and the antisense strand contains the sequence shown in SEQ ID NO.181; (23) The sense strand contains the sequence shown in SEQ ID NO.87, and the antisense strand contains the sequence shown in SEQ ID NO.185; (24) The sense strand contains the sequence shown in SEQ ID NO.90, and the antisense strand contains the sequence shown in SEQ ID NO.188; (25) The sense strand contains the sequence shown in SEQ ID NO.94, and the antisense strand contains the sequence shown in SEQ ID NO.192; More preferably, the oligonucleotide is selected from any of the following combinations of sense and antisense strands: (1) The sense strand contains the sequence shown in SEQ ID NO.33, and the antisense strand contains the sequence shown in SEQ ID NO.131; (2) The sense strand contains the sequence shown in SEQ ID NO.34, and the antisense strand contains the sequence shown in SEQ ID NO.132; (3) The sense strand contains the sequence shown in SEQ ID NO.38, and the antisense strand contains the sequence shown in SEQ ID NO.136; (4) The sense strand contains the sequence shown in SEQ ID NO.43, and the antisense strand contains the sequence shown in SEQ ID NO.141; (5) The sense strand contains the sequence shown in SEQ ID NO.53, and the antisense strand contains the sequence shown in SEQ ID NO.151; (6) The sense strand comprises the sequence shown in SEQ ID NO.60, and the antisense strand comprises the sequence shown in SEQ ID NO.158; (7) The sense strand comprises the sequence shown in SEQ ID NO.61, and the antisense strand comprises the sequence shown in SEQ ID NO.159; (8) The sense strand comprises the sequence shown in SEQ ID NO.83, and the antisense strand comprises the sequence shown in SEQ ID NO.181; (9) The sense strand comprises the sequence shown in SEQ ID NO.90, and the antisense strand comprises the sequence shown in SEQ ID NO.188; 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 1-6, wherein, The oligonucleotide comprises any one of the following combinations of sense and antisense strands: (1) The sense strand comprises the sequence shown in SEQ ID NO.198, and the antisense strand comprises the sequence shown in SEQ ID NO.261; (2) The sense strand comprises the sequence shown in SEQ ID NO.199, and the antisense strand comprises the sequence shown in SEQ ID NO.262; (3) The sense strand comprises the sequence shown in SEQ ID NO.200, and the antisense strand comprises the sequence shown in SEQ ID NO.263; (4) The sense strand comprises the sequence shown in SEQ ID NO.201, and the antisense strand comprises the sequence shown in SEQ ID NO.264; (5) The sense strand comprises the sequence shown in SEQ ID NO.202, and the antisense strand comprises the sequence shown in SEQ ID NO.265; (6) The sense strand comprises the sequence shown in SEQ ID NO.203, and the antisense strand comprises the sequence shown in SEQ ID NO.266; (7) The sense strand comprises the sequence shown in SEQ ID NO.204, and the antisense strand comprises the sequence shown in SEQ ID NO.267; (8) The sense strand comprises the sequence shown in SEQ ID NO.205, and the antisense strand comprises the sequence shown in SEQ ID NO.268; (9) The sense strand comprises the sequence shown in SEQ ID NO.206, and the antisense strand comprises the sequence shown in SEQ ID NO.269; (10) The sense strand comprises the sequence shown in SEQ ID NO.207, and the antisense strand comprises the sequence shown in SEQ ID NO.270; (11) The sense strand comprises the sequence shown in SEQ ID NO.208, and the antisense strand comprises the sequence shown in SEQ ID NO.271; (12) The sense strand comprises the sequence shown in SEQ ID NO.209, and the antisense strand comprises the sequence shown in SEQ ID NO.272; (13) The sense strand contains the sequence shown in SEQ ID NO. 210, and the antisense strand contains the sequence shown in SEQ ID NO. 273; (14) The sense strand contains the sequence shown in SEQ ID NO. 211, and the antisense strand contains the sequence shown in SEQ ID NO. 274; (15) The sense strand contains the sequence shown in SEQ ID NO. 212, and the antisense strand contains the sequence shown in SEQ ID NO. 275; (16) The sense strand contains the sequence shown in SEQ ID NO. 214, and the antisense strand contains the sequence shown in SEQ ID NO. 277; (17) The sense strand contains the sequence shown in SEQ ID NO. 215, and the antisense strand contains the sequence shown in SEQ ID NO. 278; (18) The sense strand contains the sequence shown in SEQ ID NO. 216, and the antisense strand contains the sequence shown in SEQ ID NO. 279; (19) The sense strand contains the sequence shown in SEQ ID NO. 217, and the antisense strand contains the sequence shown in SEQ ID NO. 280; (20) The sense strand contains the sequence shown in SEQ ID NO. 218, and the antisense strand contains the sequence shown in SEQ ID NO. 281; (21) The sense strand contains the sequence shown in SEQ ID NO. 219, and the antisense strand contains the sequence shown in SEQ ID NO. 282; (22) The sense strand contains the sequence shown in SEQ ID NO. 220, and the antisense strand contains the sequence shown in SEQ ID NO. 283; (23) The sense strand contains the sequence shown in SEQ ID NO. 221, and the antisense strand contains the sequence shown in SEQ ID NO. 284; (24) The sense strand contains the sequence shown in SEQ ID NO. 223, and the antisense strand contains the sequence shown in SEQ ID NO. 286; (25) The sense strand contains the sequence shown in SEQ ID NO. 224, and the antisense strand contains the sequence shown in SEQ ID NO. 287; (26) The sense strand contains the sequence shown in SEQ ID NO. 225, and the antisense strand contains the sequence shown in SEQ ID NO. 288; (27) The sense strand contains the sequence shown in SEQ ID NO. 226, and the antisense strand contains the sequence shown in SEQ ID NO. 289; (28) The sense strand contains the sequence shown in SEQ ID NO. 227, and the antisense strand contains the sequence shown in SEQ ID NO. 290; (29) The sense strand contains the sequence shown in SEQ ID NO. 228, and the antisense strand contains the sequence shown in SEQ ID NO. 291; (30) The sense strand contains the sequence shown in SEQ ID NO.229, and the antisense strand contains the sequence shown in SEQ ID NO.292; (31) The sense strand contains the sequence shown in SEQ ID NO.230, and the antisense strand contains the sequence shown in SEQ ID NO.293; (32) The sense strand contains the sequence shown in SEQ ID NO.231, and the antisense strand contains the sequence shown in SEQ ID NO.294; (33) The sense strand contains the sequence shown in SEQ ID NO.232, and the antisense strand contains the sequence shown in SEQ ID NO.295; (34) The sense strand contains the sequence shown in SEQ ID NO.233, and the antisense strand contains the sequence shown in SEQ ID NO.296; (35) The sense strand contains the sequence shown in SEQ ID NO.234, and the antisense strand contains the sequence shown in SEQ ID NO.297; (36) The sense strand contains the sequence shown in SEQ ID NO.235, and the antisense strand contains the sequence shown in SEQ ID NO.298; (37) The sense strand contains the sequence shown in SEQ ID NO.236, and the antisense strand contains the sequence shown in SEQ ID NO.299; (38) The sense strand contains the sequence shown in SEQ ID NO.237, and the antisense strand contains the sequence shown in SEQ ID NO.300; (39) The sense strand contains the sequence shown in SEQ ID NO.238, and the antisense strand contains the sequence shown in SEQ ID NO.301; (40) The sense strand contains the sequence shown in SEQ ID NO.239, and the antisense strand contains the sequence shown in SEQ ID NO.302; (41) The sense strand contains the sequence shown in SEQ ID NO.240, and the antisense strand contains the sequence shown in SEQ ID NO.303; (42) The sense strand contains the sequence shown in SEQ ID NO.241, and the antisense strand contains the sequence shown in SEQ ID NO.304; (43) The sense strand contains the sequence shown in SEQ ID NO.242, and the antisense strand contains the sequence shown in SEQ ID NO.305; (44) The sense strand contains the sequence shown in SEQ ID NO.243, and the antisense strand contains the sequence shown in SEQ ID NO.306; (45) The sense strand contains the sequence shown in SEQ ID NO.244, and the antisense strand contains the sequence shown in SEQ ID NO.307; (46) The sense strand contains the sequence shown in SEQ ID NO.245, and the antisense strand contains the sequence shown in SEQ ID NO.308; (47) The sense strand contains the sequence shown in SEQ ID NO.246, and the antisense strand contains the sequence shown in SEQ ID NO.309; (48) The sense strand contains the sequence shown in SEQ ID NO.247, and the antisense strand contains the sequence shown in SEQ ID NO.310; (49) The sense strand contains the sequence shown in SEQ ID NO.248, and the antisense strand contains the sequence shown in SEQ ID NO.311; (50) The sense strand contains the sequence shown in SEQ ID NO.249, and the antisense strand contains the sequence shown in SEQ ID NO.312; (51) The sense strand contains the sequence shown in SEQ ID NO.250, and the antisense strand contains the sequence shown in SEQ ID NO.313; (52) The sense strand contains the sequence shown in SEQ ID NO.251, and the antisense strand contains the sequence shown in SEQ ID NO.314; (53) The sense strand contains the sequence shown in SEQ ID NO.252, and the antisense strand contains the sequence shown in SEQ ID NO.315; (54) The sense strand contains the sequence shown in SEQ ID NO.253, and the antisense strand contains the sequence shown in SEQ ID NO.316; (55) The sense strand contains the sequence shown in SEQ ID NO.254, and the antisense strand contains the sequence shown in SEQ ID NO.317; (56) The sense strand contains the sequence shown in SEQ ID NO.255, and the antisense strand contains the sequence shown in SEQ ID NO.318; (57) The sense strand contains the sequence shown in SEQ ID NO.256, and the antisense strand contains the sequence shown in SEQ ID NO.319; (58) The sense strand contains the sequence shown in SEQ ID NO.257, and the antisense strand contains the sequence shown in SEQ ID NO.320; (59) The sense strand contains the sequence shown in SEQ ID NO.258, and the antisense strand contains the sequence shown in SEQ ID NO.321; Preferably, the oligonucleotide is selected from any of the following combinations of sense and antisense strands: (1) The sense strand contains the sequence shown in SEQ ID NO.202, and the antisense strand contains the sequence shown in SEQ ID NO.265; (2) The sense strand contains the sequence shown in SEQ ID NO.215, and the antisense strand contains the sequence shown in SEQ ID NO.278; (3) The sense strand contains the sequence shown in SEQ ID NO.216, and the antisense strand contains the sequence shown in SEQ ID NO.279; (4) The sense strand contains the sequence shown in SEQ ID NO.219, and the antisense strand contains the sequence shown in SEQ ID NO.282; (5) The sense strand contains the sequence shown in SEQ ID NO. 224, and the antisense strand contains the sequence shown in SEQ ID NO. 287; (6) The sense strand contains the sequence shown in SEQ ID NO. 228, and the antisense strand contains the sequence shown in SEQ ID NO. 291; (7) The sense strand contains the sequence shown in SEQ ID NO. 229, and the antisense strand contains the sequence shown in SEQ ID NO. 292; (8) The sense strand contains the sequence shown in SEQ ID NO. 234, and the antisense strand contains the sequence shown in SEQ ID NO. 297; (9) The sense strand contains the sequence shown in SEQ ID NO. 235, and the antisense strand contains the sequence shown in SEQ ID NO. 298; (10) The sense strand contains the sequence shown in SEQ ID NO. 236, and the antisense strand contains the sequence shown in SEQ ID NO. 299; (11) The sense strand contains the sequence shown in SEQ ID NO. 237, and the antisense strand contains the sequence shown in SEQ ID NO. 300; (12) The sense strand contains the sequence shown in SEQ ID NO. 238, and the antisense strand contains the sequence shown in SEQ ID NO. 301; (13) The sense strand contains the sequence shown in SEQ ID NO. 239, and the antisense strand contains the sequence shown in SEQ ID NO. 302; (14) The sense strand contains the sequence shown in SEQ ID NO. 240, and the antisense strand contains the sequence shown in SEQ ID NO. 303; (15) The sense strand contains the sequence shown in SEQ ID NO. 241, and the antisense strand contains the sequence shown in SEQ ID NO. 304; (16) The sense strand contains the sequence shown in SEQ ID NO. 242, and the antisense strand contains the sequence shown in SEQ ID NO. 305; (17) The sense strand contains the sequence shown in SEQ ID NO. 243, and the antisense strand contains the sequence shown in SEQ ID NO. 306; (18) The sense strand contains the sequence shown in SEQ ID NO. 244, and the antisense strand contains the sequence shown in SEQ ID NO. 307; (19) The sense strand contains the sequence shown in SEQ ID NO. 246, and the antisense strand contains the sequence shown in SEQ ID NO. 309; (20) The sense strand contains the sequence shown in SEQ ID NO. 247, and the antisense strand contains the sequence shown in SEQ ID NO. 310; (21) The sense strand contains the sequence shown in SEQ ID NO. 248, and the antisense strand contains the sequence shown in SEQ ID NO. 311; (22) The sense strand comprises the sequence shown in SEQ ID NO.250, and the antisense strand comprises the sequence shown in SEQ ID NO.313; (23) The sense strand comprises the sequence shown in SEQ ID NO.254, and the antisense strand comprises the sequence shown in SEQ ID NO.317; (24) The sense strand comprises the sequence shown in SEQ ID NO.255, and the antisense strand comprises the sequence shown in SEQ ID NO.318; (25) The sense strand comprises the sequence shown in SEQ ID NO.257, and the antisense strand comprises the sequence shown in SEQ ID NO.320; More preferably, the oligonucleotide is selected from any of the following combinations of sense and antisense strands: (1) The sense strand comprises the sequence shown in SEQ ID NO.215, and the antisense strand comprises the sequence shown in SEQ ID NO.278; (2) The sense strand comprises the sequence shown in SEQ ID NO.216, and the antisense strand comprises the sequence shown in SEQ ID NO.279; (3) The sense strand comprises the sequence shown in SEQ ID NO.219, and the antisense strand comprises the sequence shown in SEQ ID NO.282; (4) The sense strand comprises the sequence shown in SEQ ID NO.224, and the antisense strand comprises the sequence shown in SEQ ID NO.287; (5) The sense strand comprises the sequence shown in SEQ ID NO.229, and the antisense strand comprises the sequence shown in SEQ ID NO.292; (6) The sense strand comprises the sequence shown in SEQ ID NO.235, and the antisense strand comprises the sequence shown in SEQ ID NO.298; (7) The sense strand comprises the sequence shown in SEQ ID NO.236, and the antisense strand comprises the sequence shown in SEQ ID NO.299; (8) The sense strand comprises the sequence shown in SEQ ID NO.250, and the antisense strand comprises the sequence shown in SEQ ID NO.313; (9) The sense strand comprises the sequence shown in SEQ ID NO.255, and the antisense strand comprises the sequence shown in SEQ ID NO.318; wherein each strand is independently 19 to 25 nucleotides in length.

8. A conjugate or a pharmaceutically acceptable salt thereof for reducing the expression of the HSD17B13 gene, comprising: (i) an oligonucleotide as described in any one of claims 1-7 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; 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 and (NAG37)s; 9. A composition comprising the oligonucleotide according to any one of claims 1-7 or a pharmaceutically acceptable salt thereof, or the conjugate according to claim 8 or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier; Preferably, the dosage form of the composition is an oral preparation, an intravenous injection, a subcutaneous injection or an intramuscular injection; Preferably, the combination further comprises other drugs for treating and / or preventing HSD17B13-related diseases; Preferably, the HSD17B13-related diseases are selected from non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, cirrhosis, etc., and complications related to these diseases such as type 2 diabetes, cardiovascular diseases, chronic kidney diseases, etc.

10. Use of the oligonucleotide according to any one of claims 1-7 or a pharmaceutically acceptable salt thereof, the conjugate according to claim 8 or the composition according to claim 9 in the preparation of a drug for treating and / or preventing HSD17B13-related diseases; Preferably, the HSD17B13-related diseases are selected from non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, cirrhosis, etc., and complications related to these diseases such as type 2 diabetes, cardiovascular diseases, chronic kidney diseases, etc.; More preferably, the HSD17B13-related diseases are selected from non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, cirrhosis, etc.