WO2024027518A1 - Double-stranded ribonucleic acid for inhibiting angptl3 gene expression, and modifier, conjugate, and use thereof - Google Patents

Double-stranded ribonucleic acid for inhibiting angptl3 gene expression, and modifier, conjugate, and use thereof Download PDF

Info

Publication number
WO2024027518A1
WO2024027518A1 PCT/CN2023/108634 CN2023108634W WO2024027518A1 WO 2024027518 A1 WO2024027518 A1 WO 2024027518A1 CN 2023108634 W CN2023108634 W CN 2023108634W WO 2024027518 A1 WO2024027518 A1 WO 2024027518A1
Authority
WO
WIPO (PCT)
Prior art keywords
nucleotide sequence
seq
sequence shown
strand includes
antisense strand
Prior art date
Application number
PCT/CN2023/108634
Other languages
French (fr)
Chinese (zh)
Inventor
王书成
黄河
王岩
林国良
产运霞
耿玉先
汪小君
荣梅
Original Assignee
北京福元医药股份有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 北京福元医药股份有限公司 filed Critical 北京福元医药股份有限公司
Publication of WO2024027518A1 publication Critical patent/WO2024027518A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1136—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against growth factors, growth regulators, cytokines, lymphokines or hormones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/7125—Nucleic acids or oligonucleotides having modified internucleoside linkage, i.e. other than 3'-5' phosphodiesters
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/543—Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
    • A61K47/544—Phospholipids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/02—Nutrients, e.g. vitamins, minerals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • C07H21/02—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with ribosyl as saccharide radical
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/14—Type of nucleic acid interfering N.A.
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/31—Chemical structure of the backbone
    • C12N2310/315—Phosphorothioates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/32—Chemical structure of the sugar
    • C12N2310/321—2'-O-R Modification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/32—Chemical structure of the sugar
    • C12N2310/322—2'-R Modification

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Diabetes (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Hematology (AREA)
  • Obesity (AREA)
  • Biochemistry (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Epidemiology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Rheumatology (AREA)
  • Endocrinology (AREA)
  • Biophysics (AREA)
  • Emergency Medicine (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Nutrition Science (AREA)
  • Immunology (AREA)
  • Physics & Mathematics (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Cardiology (AREA)
  • Plant Pathology (AREA)

Abstract

The present disclosure relates to a double-stranded ribonucleic acid for inhibiting ANGPTL3 gene expression, and a modifier, a conjugate, and use thereof. Specifically, the present disclosure relates to a double-stranded ribonucleic acid, a double-stranded ribonucleic acid modifier, a double-stranded ribonucleic acid conjugate, a pharmaceutical composition and use for inhibiting ANGPTL3 gene expression, and a method for inhibiting intracellular ANGPTL3 gene expression. The double-stranded ribonucleic acid provided in the present disclosure can bind in cells to form an RNA-induced silencing complex (RISC), which cleaves the mRNA transcribed by the ANGPTL3 gene, efficiently and specifically inhibiting the expression of the ANGPTL3 gene. The double-stranded ribonucleic acid is used for treating diseases associated with ANGPTL3 gene-mediated metabolic disorders, and has important application prospects in clinical disease treatment.

Description

用于抑制ANGPTL3基因表达的双链核糖核酸及其修饰物、缀合物和用途Double-stranded ribonucleic acid for inhibiting ANGPTL3 gene expression and its modifications, conjugates and uses
优先权和相关申请Priority and related applications
本公开要求于2022年8月4日提交中国专利局、申请号为202210935615.8、发明名称为“用于抑制ANGPTL3基因表达的双链核糖核酸及其修饰物、缀合物和用途”的中国专利申请的优先权，其全部内容通过引用结合在本公开中。This disclosure requires a Chinese patent application submitted to the China Patent Office on August 4, 2022, with the application number 202210935615.8 and the invention title "Double-stranded ribonucleic acid for inhibiting ANGPTL3 gene expression and its modifications, conjugates and uses" priority, the entire contents of which are incorporated by reference into this disclosure.
技术领域Technical field
本公开属于生物医药领域，具体来说，本公开涉及一种用于抑制ANGTPL3基因表达的双链核糖核酸、双链核糖核酸修饰物、双链核糖核酸缀合物、药物组合物和用途，以及用于抑制细胞内ANGTPL3基因表达的方法。The present disclosure belongs to the field of biomedicine. Specifically, the present disclosure relates to a double-stranded ribonucleic acid, a double-stranded ribonucleic acid modification, a double-stranded ribonucleic acid conjugate, a pharmaceutical composition and uses for inhibiting ANGTPL3 gene expression, and Methods for inhibiting intracellular ANGTPL3 gene expression.
背景技术Background technique
血管生成素-样蛋白3(ANGPTL3)，亦称为ANGPL3或ANG3，是一种分泌型糖蛋白，属于分泌调控脂质代谢的因子的类血管生成素家族的成员。它主要在肝脏中合成，并且随后分泌到循环中，最终主要在肝脏中表达。ANGPTL3通过与脂肪组织结合，抑制脂蛋白脂肪酶的活性来调节脂质代谢，具体表现为抑制脂蛋白脂肪酶和内皮脂肪酶的催化活性。而这两类酶的抑制可导致甘油三酯、高密度脂蛋白(HDL)和磷脂的血浆水平增加，因此，ANGPTL3的低表达可以减缓血脂异常的各类疾病，包括乳糜微粒血症综合征、2型糖尿病、家族性部分脂肪营养不良、高胆固醇血症、家族性高胆固醇血症、高甘油三酯血症、非酒精性脂肪肝、动脉粥样硬化、高甘油三酯血症、膝盖损伤及骨关节炎、血脂异常及相关代谢疾病。Angiopoietin-like protein 3 (ANGPTL3), also known as ANGPL3 or ANG3, is a secreted glycoprotein and a member of the angiopoietin-like family that secretes factors that regulate lipid metabolism. It is primarily synthesized in the liver and subsequently secreted into the circulation, where it is ultimately expressed primarily in the liver. ANGPTL3 regulates lipid metabolism by binding to adipose tissue and inhibiting the activity of lipoprotein lipase, specifically by inhibiting the catalytic activity of lipoprotein lipase and endothelial lipase. Inhibition of these two enzymes can lead to increased plasma levels of triglycerides, high-density lipoprotein (HDL), and phospholipids. Therefore, low expression of ANGPTL3 can slow down various diseases with dyslipidemia, including chylomicronemia syndrome, Type 2 diabetes, familial partial lipodystrophy, hypercholesterolemia, familial hypercholesterolemia, hypertriglyceridemia, non-alcoholic fatty liver disease, atherosclerosis, hypertriglyceridemia, knee injury and osteoarthritis, dyslipidemia and related metabolic diseases.
血脂异常，又名高脂血症，是脂肪代谢或运转异常，使血浆脂质高于正常值的一种全身性疾病，脂质代谢障碍可导致血清脂质，诸如三酸甘油酯和/或胆固醇的水平升高。血清脂质升高与高血压、心血管疾病、糖尿病及其他病理状况强烈相关。高三酸甘油酯血症为脂质代谢障碍的实例，其特征为三酸甘油酯的高血液水平，这些已经不再罕见，正严重威胁着全球患者的健康。目前脂质代谢障碍的现有治疗包括节食、运动、或使用他汀类(statins)及其他药物治疗，但并不能满足大多数患者的健康需求。因此，开发新型的ANGPTL3基因的表达抑制剂，是目前亟需解决的重要问题。Dyslipidemia, also known as hyperlipidemia, is a systemic disease in which fat metabolism or operation is abnormal, causing plasma lipids to be higher than normal. Lipid metabolism disorders can cause serum lipids, such as triglycerides and/or Increased cholesterol levels. Elevated serum lipids are strongly associated with hypertension, cardiovascular disease, diabetes, and other pathological conditions. Hypertriglyceridemia is an example of a lipid metabolism disorder characterized by high blood levels of triglycerides. These are no longer uncommon and are seriously threatening the health of patients worldwide. Currently, existing treatments for lipid metabolism disorders include diet, exercise, or the use of statins and other drugs, but they cannot meet the health needs of most patients. Therefore, the development of new inhibitors of ANGPTL3 gene expression is an important issue that needs to be solved urgently.
发明内容Contents of the invention
发明要解决的问题Invent the problem to be solved
鉴于现有技术中存在的问题，例如，需要开发更多ANGPTL3抑制剂，用于治疗包括脂质代谢异常在内的ANGPTL3相关疾病。本公开旨在提供一系列用于抑制ANGPTL3基因表达的双链核糖核酸、双链核糖核酸修饰物、双链核糖核酸缀合物及药物组合物，能够抑制ANGPTL3基因表达，在临床疾病治疗中具有重要应用前景。In view of the problems existing in the existing technology, for example, more ANGPTL3 inhibitors need to be developed for the treatment of ANGPTL3-related diseases including abnormal lipid metabolism. The present disclosure aims to provide a series of double-stranded ribonucleic acids, double-stranded ribonucleic acid modifications, double-stranded ribonucleic acid conjugates and pharmaceutical compositions for inhibiting ANGPTL3 gene expression, which can inhibit ANGPTL3 gene expression and have utility in clinical disease treatment. important application prospects.
用于解决问题的方案solutions to problems
[1].一种双链核糖核酸，所述双链核糖核酸包括正义链和反义链，所述正义链与所述反义链互补和/或基本上反向互补形成所述双链核糖核酸的双链区；[1]. A double-stranded ribonucleic acid, the double-stranded ribonucleic acid includes a sense strand and an antisense strand, the sense strand is complementary and/or substantially reverse complementary to the antisense strand to form the double-stranded ribonucleic acid The double-stranded region of a nucleic acid;
其中，所述正义链包含与靶标序列中至少15个连续核苷酸的差异不超过3个核苷酸的序列A，所述反义链包含与靶标序列中至少15个连续核苷酸的反向互补序列的差异不超过3个核苷酸的序列B；Wherein, the sense strand includes a sequence A that differs from at least 15 consecutive nucleotides in the target sequence by no more than 3 nucleotides, and the antisense strand includes a sequence A that differs from at least 15 consecutive nucleotides in the target sequence. Sequence B that differs from the complementary sequence by no more than 3 nucleotides;
所述靶标序列选自如SEQ ID NO:1-7任一项所示的核苷酸序列。The target sequence is selected from the nucleotide sequence shown in any one of SEQ ID NO: 1-7.
[2].根据[1]所述的双链核糖核酸，其中，所述靶标序列选自如SEQ ID NO:8-25任一 项所示的核苷酸序列，所述正义链包含与SEQ ID NO:8-25任一项所示的核苷酸序列中至少15个连续核苷酸组成的序列相比差异不超过1个核苷酸的序列A，所述反义链包含与SEQ ID NO:8-25任一项所示的核苷酸序列中至少15个连续核苷酸组成的序列的反向互补序列相比差异不超过1个核苷酸的序列B；[2]. The double-stranded ribonucleic acid according to [1], wherein the target sequence is selected from any one of SEQ ID NO: 8-25 The nucleotide sequence shown in the item, the sense strand contains a sequence consisting of at least 15 consecutive nucleotides in the nucleotide sequence shown in any one of SEQ ID NO: 8-25, which differs by no more than 1 Sequence A of nucleotides, the antisense strand comprising a difference compared with the reverse complementary sequence of a sequence consisting of at least 15 consecutive nucleotides in the nucleotide sequence shown in any one of SEQ ID NO:8-25 Sequence B of not more than 1 nucleotide;
可选地，所述差异的核苷酸位于所述序列A的3’末端；和/或，所述差异的核苷酸位于所述序列B的3’末端。Optionally, the different nucleotides are located at the 3' end of the sequence A; and/or the different nucleotides are located at the 3' end of the sequence B.
[3].根据[1]或[2]所述的双链核糖核酸，其中，所述正义链由15-28个核苷酸组成，优选19-25个核苷酸，更优选19-23个核苷酸，更优选19、21或23个核苷酸。[3]. The double-stranded ribonucleic acid according to [1] or [2], wherein the sense strand consists of 15-28 nucleotides, preferably 19-25 nucleotides, more preferably 19-23 nucleotides nucleotides, more preferably 19, 21 or 23 nucleotides.
[4].根据[3]所述的双链核糖核酸，其中，所述正义链的核苷酸序列为与SEQ ID NO:8-25任一项所示的核苷酸序列中的15-28个连续核苷酸组成的序列相比差异不超过1个核苷酸的序列A，优选19-25个连续核苷酸，更优选19-23个连续核苷酸，更优选19、21或23个核苷酸。[4]. The double-stranded ribonucleic acid according to [3], wherein the nucleotide sequence of the sense strand is 15-15 of the nucleotide sequence shown in any one of SEQ ID NO:8-25. Compared with sequence A that differs by no more than 1 nucleotide, a sequence composed of 28 consecutive nucleotides is preferably 19-25 consecutive nucleotides, more preferably 19-23 consecutive nucleotides, and more preferably 19, 21 or 23 nucleotides.
[5].根据[1]-[4]任一项所述的双链核糖核酸，其中，所述反义链由15-28个核苷酸组成，优选19-25个核苷酸，更优选19-23个核苷酸，更优选19、21或23个核苷酸。[5]. The double-stranded ribonucleic acid according to any one of [1]-[4], wherein the antisense strand consists of 15-28 nucleotides, preferably 19-25 nucleotides, more Preferred are 19-23 nucleotides, more preferably 19, 21 or 23 nucleotides.
[6].根据[5]所述的双链核糖核酸，其中，所述反义链的核苷酸序列是与SEQ ID NO:8-25任一项所示的核苷酸序列中的15-28个连续核苷酸组成的序列的反向互补序列相比差异不超过1个核苷酸的序列B，优选19-25个连续核苷酸，更优选19-23个连续核苷酸，更优选19、21或23个核苷酸。[6]. The double-stranded ribonucleic acid according to [5], wherein the nucleotide sequence of the antisense strand is 15 nucleotides from the nucleotide sequence shown in any one of SEQ ID NO: 8-25 -The reverse complement of a sequence consisting of 28 consecutive nucleotides is preferably 19-25 consecutive nucleotides, more preferably 19-23 consecutive nucleotides, compared to sequence B which differs by no more than 1 nucleotide, More preferred are 19, 21 or 23 nucleotides.
[7].根据[1]-[6]任一项所述的双链核糖核酸，其中，所述双链区的长度为15-25个核苷酸，优选19-23个核苷酸，更优选19-21个核苷酸，更优选19、21或23个核苷酸。[7]. The double-stranded ribonucleic acid according to any one of [1]-[6], wherein the length of the double-stranded region is 15-25 nucleotides, preferably 19-23 nucleotides, More preferably 19-21 nucleotides, more preferably 19, 21 or 23 nucleotides.
[8].根据[1]-[7]任一项所述的双链核糖核酸，其中，[8]. The double-stranded ribonucleic acid according to any one of [1]-[7], wherein,
所述正义链与所述反义链互补形成所述双链区，且所述正义链的3’末端具有1-2个延伸出所述双链区的突出的核苷酸，所述反义链的3’末端形成平末端；或者，The sense strand and the antisense strand are complementary to form the double-stranded region, and the 3' end of the sense strand has 1-2 protruding nucleotides extending out of the double-stranded region, and the antisense strand is complementary to the double-stranded region. The 3' end of the chain forms a blunt end; or,
所述正义链与所述反义链互补形成所述双链区，且所述反义链的3’末端具有1-2个延伸出所述双链区的突出的核苷酸，所述正义链的3’末端形成平末端；或者，The sense strand and the antisense strand are complementary to form the double-stranded region, and the 3' end of the antisense strand has 1-2 protruding nucleotides extending out of the double-stranded region, and the sense strand The 3' end of the chain forms a blunt end; or,
所述正义链与所述反义链互补形成所述双链区，且所述正义链与所述反义链的3’末端均具有1-2个延伸出所述双链区的突出的核苷酸；或者，The sense strand and the antisense strand are complementary to form the double-stranded region, and the 3' ends of the sense strand and the antisense strand each have 1-2 protruding cores extending out of the double-stranded region. glycoside; or,
所述正义链与所述反义链互补形成所述双链区，且所述正义链与所述反义链的3’末端均形成平末端。The sense strand and the antisense strand are complementary to form the double-stranded region, and the 3' ends of the sense strand and the antisense strand both form blunt ends.
[9].根据[1]-[8]任一项所述的双链核糖核酸，其中，所述正义链与所述反义链选自如下组合：[9]. The double-stranded ribonucleic acid according to any one of [1]-[8], wherein the sense strand and the antisense strand are selected from the following combinations:
1)所述正义链包含如SEQ ID NO:26所示的核苷酸序列，所述反义链包含如SEQ ID NO:121所示的核苷酸序列；1) The sense strand includes the nucleotide sequence shown in SEQ ID NO:26, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:121;
2)所述正义链包含如SEQ ID NO:27所示的核苷酸序列，所述反义链包含如SEQ ID NO:122所示的核苷酸序列；2) The sense strand includes the nucleotide sequence shown in SEQ ID NO:27, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:122;
3)所述正义链包含如SEQ ID NO:28所示的核苷酸序列，所述反义链包含如SEQ ID NO:123所示的核苷酸序列；3) The sense strand includes the nucleotide sequence shown in SEQ ID NO:28, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:123;
4)所述正义链包含如SEQ ID NO:29所示的核苷酸序列，所述反义链包含如SEQ ID NO:124所示的核苷酸序列；4) The sense strand includes the nucleotide sequence shown in SEQ ID NO:29, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:124;
5)所述正义链包含如SEQ ID NO:30所示的核苷酸序列，所述反义链包含如SEQ ID NO:125所示的核苷酸序列；5) The sense strand includes the nucleotide sequence shown in SEQ ID NO:30, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:125;
6)所述正义链包含如SEQ ID NO:31所示的核苷酸序列，所述反义链包含如SEQ ID NO:126所示的核苷酸序列；6) The sense strand includes the nucleotide sequence shown in SEQ ID NO:31, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:126;
7)所述正义链包含如SEQ ID NO:32所示的核苷酸序列，所述反义链包含如SEQ ID NO:127所示的核苷酸序列； 7) The sense strand includes the nucleotide sequence shown in SEQ ID NO:32, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:127;
8)所述正义链包含如SEQ ID NO:33所示的核苷酸序列，所述反义链包含如SEQ ID NO:128所示的核苷酸序列；8) The sense strand includes the nucleotide sequence shown in SEQ ID NO:33, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:128;
9)所述正义链包含如SEQ ID NO:34所示的核苷酸序列，所述反义链包含如SEQ ID NO:129所示的核苷酸序列；9) The sense strand includes the nucleotide sequence shown in SEQ ID NO:34, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:129;
10)所述正义链包含如SEQ ID NO:35所示的核苷酸序列，所述反义链包含如SEQ ID NO:130所示的核苷酸序列；10) The sense strand includes the nucleotide sequence shown in SEQ ID NO:35, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:130;
11)所述正义链包含如SEQ ID NO:36所示的核苷酸序列，所述反义链包含如SEQ ID NO:131所示的核苷酸序列；11) The sense strand includes the nucleotide sequence shown in SEQ ID NO:36, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:131;
12)所述正义链包含如SEQ ID NO:37所示的核苷酸序列，所述反义链包含如SEQ ID NO:132所示的核苷酸序列；12) The sense strand includes the nucleotide sequence shown in SEQ ID NO:37, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:132;
13)所述正义链包含如SEQ ID NO:38所示的核苷酸序列，所述反义链包含如SEQ ID NO:133所示的核苷酸序列；13) The sense strand includes the nucleotide sequence shown in SEQ ID NO:38, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:133;
14)所述正义链包含如SEQ ID NO:39所示的核苷酸序列，所述反义链包含如SEQ ID NO:134所示的核苷酸序列；14) The sense strand includes the nucleotide sequence shown in SEQ ID NO:39, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:134;
15)所述正义链包含如SEQ ID NO:40所示的核苷酸序列，所述反义链包含如SEQ ID NO:135所示的核苷酸序列；15) The sense strand includes the nucleotide sequence shown in SEQ ID NO:40, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:135;
16)所述正义链包含如SEQ ID NO:41所示的核苷酸序列，所述反义链包含如SEQ ID NO:136所示的核苷酸序列；16) The sense strand includes the nucleotide sequence shown in SEQ ID NO:41, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:136;
17)所述正义链包含如SEQ ID NO:42所示的核苷酸序列，所述反义链包含如SEQ ID NO:137所示的核苷酸序列；17) The sense strand includes the nucleotide sequence shown in SEQ ID NO:42, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:137;
18)所述正义链包含如SEQ ID NO:43所示的核苷酸序列，所述反义链包含如SEQ ID NO:138所示的核苷酸序列；18) The sense strand includes the nucleotide sequence shown in SEQ ID NO:43, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:138;
19)所述正义链包含如SEQ ID NO:44所示的核苷酸序列，所述反义链包含如SEQ ID NO:139所示的核苷酸序列；19) The sense strand includes the nucleotide sequence shown in SEQ ID NO:44, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:139;
20)所述正义链包含如SEQ ID NO:45所示的核苷酸序列，所述反义链包含如SEQ ID NO:140所示的核苷酸序列；20) The sense strand includes the nucleotide sequence shown in SEQ ID NO:45, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:140;
21)所述正义链包含如SEQ ID NO:46所示的核苷酸序列，所述反义链包含如SEQ ID NO:141所示的核苷酸序列；21) The sense strand includes the nucleotide sequence shown in SEQ ID NO:46, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:141;
22)所述正义链包含如SEQ ID NO:47所示的核苷酸序列，所述反义链包含如SEQ ID NO:142所示的核苷酸序列；22) The sense strand includes the nucleotide sequence shown in SEQ ID NO:47, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:142;
23)所述正义链包含如SEQ ID NO:48所示的核苷酸序列，所述反义链包含如SEQ ID NO:143所示的核苷酸序列；23) The sense strand includes the nucleotide sequence shown in SEQ ID NO:48, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:143;
24)所述正义链包含如SEQ ID NO:49所示的核苷酸序列，所述反义链包含如SEQ ID NO:144所示的核苷酸序列；24) The sense strand includes the nucleotide sequence shown in SEQ ID NO:49, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:144;
25)所述正义链包含如SEQ ID NO:50所示的核苷酸序列，所述反义链包含如SEQ ID NO:145所示的核苷酸序列；25) The sense strand includes the nucleotide sequence shown in SEQ ID NO:50, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:145;
26)所述正义链包含如SEQ ID NO:51所示的核苷酸序列，所述反义链包含如SEQ ID NO:146所示的核苷酸序列；26) The sense strand includes the nucleotide sequence shown in SEQ ID NO:51, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:146;
27)所述正义链包含如SEQ ID NO:52所示的核苷酸序列，所述反义链包含如SEQ ID NO:147所示的核苷酸序列；27) The sense strand includes the nucleotide sequence shown in SEQ ID NO:52, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:147;
28)所述正义链包含如SEQ ID NO:53所示的核苷酸序列，所述反义链包含如SEQ ID NO:148所示的核苷酸序列；28) The sense strand includes the nucleotide sequence shown in SEQ ID NO:53, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:148;
29)所述正义链包含如SEQ ID NO:54所示的核苷酸序列，所述反义链包含如SEQ ID NO:149所示的核苷酸序列；29) The sense strand includes the nucleotide sequence shown in SEQ ID NO:54, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:149;
30)所述正义链包含如SEQ ID NO:55所示的核苷酸序列，所述反义链包含如SEQ ID  NO:150所示的核苷酸序列；30) The sense strand includes the nucleotide sequence shown in SEQ ID NO:55, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO. The nucleotide sequence shown in NO:150;
31)所述正义链包含如SEQ ID NO:56所示的核苷酸序列，所述反义链包含如SEQ ID NO:151所示的核苷酸序列；31) The sense strand includes the nucleotide sequence shown in SEQ ID NO:56, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:151;
32)所述正义链包含如SEQ ID NO:57所示的核苷酸序列，所述反义链包含如SEQ ID NO:152所示的核苷酸序列；32) The sense strand includes the nucleotide sequence shown in SEQ ID NO:57, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:152;
33)所述正义链包含如SEQ ID NO:58所示的核苷酸序列，所述反义链包含如SEQ ID NO:153所示的核苷酸序列；33) The sense strand includes the nucleotide sequence shown in SEQ ID NO:58, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:153;
34)所述正义链包含如SEQ ID NO:59所示的核苷酸序列，所述反义链包含如SEQ ID NO:154所示的核苷酸序列；34) The sense strand includes the nucleotide sequence shown in SEQ ID NO:59, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:154;
35)所述正义链包含如SEQ ID NO:60所示的核苷酸序列，所述反义链包含如SEQ ID NO:155所示的核苷酸序列；35) The sense strand includes the nucleotide sequence shown in SEQ ID NO:60, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:155;
36)所述正义链包含如SEQ ID NO:61所示的核苷酸序列，所述反义链包含如SEQ ID NO:156所示的核苷酸序列；36) The sense strand includes the nucleotide sequence shown in SEQ ID NO:61, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:156;
37)所述正义链包含如SEQ ID NO:62所示的核苷酸序列，所述反义链包含如SEQ ID NO:157所示的核苷酸序列；37) The sense strand includes the nucleotide sequence shown in SEQ ID NO:62, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:157;
38)所述正义链包含如SEQ ID NO:63所示的核苷酸序列，所述反义链包含如SEQ ID NO:158所示的核苷酸序列；38) The sense strand includes the nucleotide sequence shown in SEQ ID NO: 63, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 158;
39)所述正义链包含如SEQ ID NO:64所示的核苷酸序列，所述反义链包含如SEQ ID NO:159所示的核苷酸序列；39) The sense strand includes the nucleotide sequence shown in SEQ ID NO:64, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:159;
40)所述正义链包含如SEQ ID NO:65所示的核苷酸序列，所述反义链包含如SEQ ID NO:160所示的核苷酸序列；40) The sense strand includes the nucleotide sequence shown in SEQ ID NO:65, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:160;
41)所述正义链包含如SEQ ID NO:66所示的核苷酸序列，所述反义链包含如SEQ ID NO:161所示的核苷酸序列；41) The sense strand includes the nucleotide sequence shown in SEQ ID NO:66, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:161;
42)所述正义链包含如SEQ ID NO:67所示的核苷酸序列，所述反义链包含如SEQ ID NO:162所示的核苷酸序列；42) The sense strand includes the nucleotide sequence shown in SEQ ID NO:67, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:162;
43)所述正义链包含如SEQ ID NO:68所示的核苷酸序列，所述反义链包含如SEQ ID NO:163所示的核苷酸序列；43) The sense strand includes the nucleotide sequence shown in SEQ ID NO:68, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:163;
44)所述正义链包含如SEQ ID NO:69所示的核苷酸序列，所述反义链包含如SEQ ID NO:164所示的核苷酸序列；44) The sense strand includes the nucleotide sequence shown in SEQ ID NO:69, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:164;
45)所述正义链包含如SEQ ID NO:70所示的核苷酸序列，所述反义链包含如SEQ ID NO:165所示的核苷酸序列；45) The sense strand includes the nucleotide sequence shown in SEQ ID NO:70, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:165;
46)所述正义链包含如SEQ ID NO:71所示的核苷酸序列，所述反义链包含如SEQ ID NO:166所示的核苷酸序列；46) The sense strand includes the nucleotide sequence shown in SEQ ID NO:71, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:166;
47)所述正义链包含如SEQ ID NO:72所示的核苷酸序列，所述反义链包含如SEQ ID NO:167所示的核苷酸序列；47) The sense strand includes the nucleotide sequence shown in SEQ ID NO:72, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:167;
48)所述正义链包含如SEQ ID NO:73所示的核苷酸序列，所述反义链包含如SEQ ID NO:168所示的核苷酸序列；48) The sense strand includes the nucleotide sequence shown in SEQ ID NO:73, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:168;
49)所述正义链包含如SEQ ID NO:74所示的核苷酸序列，所述反义链包含如SEQ ID NO:169所示的核苷酸序列；49) The sense strand includes the nucleotide sequence shown in SEQ ID NO:74, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:169;
50)所述正义链包含如SEQ ID NO:75所示的核苷酸序列，所述反义链包含如SEQ ID NO:170所示的核苷酸序列；50) The sense strand includes the nucleotide sequence shown in SEQ ID NO:75, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:170;
51)所述正义链包含如SEQ ID NO:76所示的核苷酸序列，所述反义链包含如SEQ ID NO:171所示的核苷酸序列；51) The sense strand includes the nucleotide sequence shown in SEQ ID NO:76, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:171;
52)所述正义链包含如SEQ ID NO:77所示的核苷酸序列，所述反义链包含如SEQ ID NO:172所示的核苷酸序列； 52) The sense strand includes the nucleotide sequence shown in SEQ ID NO:77, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:172;
53)所述正义链包含如SEQ ID NO:78所示的核苷酸序列，所述反义链包含如SEQ ID NO:173所示的核苷酸序列；53) The sense strand includes the nucleotide sequence shown in SEQ ID NO:78, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:173;
54)所述正义链包含如SEQ ID NO:79所示的核苷酸序列，所述反义链包含如SEQ ID NO:174所示的核苷酸序列；54) The sense strand includes the nucleotide sequence shown in SEQ ID NO:79, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:174;
55)所述正义链包含如SEQ ID NO:80所示的核苷酸序列，所述反义链包含如SEQ ID NO:175所示的核苷酸序列；55) The sense strand includes the nucleotide sequence shown in SEQ ID NO:80, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:175;
56)所述正义链包含如SEQ ID NO:81所示的核苷酸序列，所述反义链包含如SEQ ID NO:176所示的核苷酸序列；56) The sense strand includes the nucleotide sequence shown in SEQ ID NO:81, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:176;
57)所述正义链包含如SEQ ID NO:82所示的核苷酸序列，所述反义链包含如SEQ ID NO:177所示的核苷酸序列；57) The sense strand includes the nucleotide sequence shown in SEQ ID NO:82, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:177;
58)所述正义链包含如SEQ ID NO:83所示的核苷酸序列，所述反义链包含如SEQ ID NO:178所示的核苷酸序列；58) The sense strand includes the nucleotide sequence shown in SEQ ID NO:83, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:178;
59)所述正义链包含如SEQ ID NO:84所示的核苷酸序列，所述反义链包含如SEQ ID NO:179所示的核苷酸序列；59) The sense strand includes the nucleotide sequence shown in SEQ ID NO:84, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:179;
60)所述正义链包含如SEQ ID NO:85所示的核苷酸序列，所述反义链包含如SEQ ID NO:180所示的核苷酸序列；60) The sense strand includes the nucleotide sequence shown in SEQ ID NO:85, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:180;
61)所述正义链包含如SEQ ID NO:86所示的核苷酸序列，所述反义链包含如SEQ ID NO:181所示的核苷酸序列；61) The sense strand includes the nucleotide sequence shown in SEQ ID NO:86, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:181;
62)所述正义链包含如SEQ ID NO:87所示的核苷酸序列，所述反义链包含如SEQ ID NO:182所示的核苷酸序列；62) The sense strand includes the nucleotide sequence shown in SEQ ID NO:87, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:182;
63)所述正义链包含如SEQ ID NO:88所示的核苷酸序列，所述反义链包含如SEQ ID NO:183所示的核苷酸序列；63) The sense strand includes the nucleotide sequence shown in SEQ ID NO:88, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:183;
64)所述正义链包含如SEQ ID NO:89所示的核苷酸序列，所述反义链包含如SEQ ID NO:184所示的核苷酸序列；64) The sense strand includes the nucleotide sequence shown in SEQ ID NO:89, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:184;
65)所述正义链包含如SEQ ID NO:90所示的核苷酸序列，所述反义链包含如SEQ ID NO:185所示的核苷酸序列；65) The sense strand includes the nucleotide sequence shown in SEQ ID NO:90, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:185;
66)所述正义链包含如SEQ ID NO:91所示的核苷酸序列，所述反义链包含如SEQ ID NO:186所示的核苷酸序列；66) The sense strand includes the nucleotide sequence shown in SEQ ID NO:91, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:186;
67)所述正义链包含如SEQ ID NO:92所示的核苷酸序列，所述反义链包含如SEQ ID NO:187所示的核苷酸序列；67) The sense strand includes the nucleotide sequence shown in SEQ ID NO:92, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:187;
68)所述正义链包含如SEQ ID NO:93所示的核苷酸序列，所述反义链包含如SEQ ID NO:188所示的核苷酸序列；68) The sense strand includes the nucleotide sequence shown in SEQ ID NO:93, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:188;
69)所述正义链包含如SEQ ID NO:94所示的核苷酸序列，所述反义链包含如SEQ ID NO:189所示的核苷酸序列；69) The sense strand includes the nucleotide sequence shown in SEQ ID NO:94, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:189;
70)所述正义链包含如SEQ ID NO:95所示的核苷酸序列，所述反义链包含如SEQ ID NO:190所示的核苷酸序列；70) The sense strand includes the nucleotide sequence shown in SEQ ID NO:95, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:190;
71)所述正义链包含如SEQ ID NO:96所示的核苷酸序列，所述反义链包含如SEQ ID NO:191所示的核苷酸序列；71) The sense strand includes the nucleotide sequence shown in SEQ ID NO:96, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:191;
72)所述正义链包含如SEQ ID NO:97所示的核苷酸序列，所述反义链包含如SEQ ID NO:192所示的核苷酸序列；72) The sense strand includes the nucleotide sequence shown in SEQ ID NO:97, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:192;
73)所述正义链包含如SEQ ID NO:98所示的核苷酸序列，所述反义链包含如SEQ ID NO:193所示的核苷酸序列；73) The sense strand includes the nucleotide sequence shown in SEQ ID NO:98, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:193;
74)所述正义链包含如SEQ ID NO:99所示的核苷酸序列，所述反义链包含如SEQ ID NO:194所示的核苷酸序列；74) The sense strand includes the nucleotide sequence shown in SEQ ID NO:99, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:194;
75)所述正义链包含如SEQ ID NO:100所示的核苷酸序列，所述反义链包含如SEQ ID  NO:195所示的核苷酸序列；75) The sense strand includes the nucleotide sequence shown in SEQ ID NO:100, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:100 The nucleotide sequence shown in NO:195;
76)所述正义链包含如SEQ ID NO:101所示的核苷酸序列，所述反义链包含如SEQ ID NO:196所示的核苷酸序列；76) The sense strand includes the nucleotide sequence shown in SEQ ID NO:101, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:196;
77)所述正义链包含如SEQ ID NO:102所示的核苷酸序列，所述反义链包含如SEQ ID NO:197所示的核苷酸序列；77) The sense strand includes the nucleotide sequence shown in SEQ ID NO:102, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:197;
78)所述正义链包含如SEQ ID NO:103所示的核苷酸序列，所述反义链包含如SEQ ID NO:198所示的核苷酸序列；78) The sense strand includes the nucleotide sequence shown in SEQ ID NO:103, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:198;
79)所述正义链包含如SEQ ID NO:104所示的核苷酸序列，所述反义链包含如SEQ ID NO:199所示的核苷酸序列；79) The sense strand includes the nucleotide sequence shown in SEQ ID NO:104, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:199;
80)所述正义链包含如SEQ ID NO:105所示的核苷酸序列，所述反义链包含如SEQ ID NO:200所示的核苷酸序列；80) The sense strand includes the nucleotide sequence shown in SEQ ID NO:105, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:200;
81)所述正义链包含如SEQ ID NO:106所示的核苷酸序列，所述反义链包含如SEQ ID NO:201所示的核苷酸序列；81) The sense strand includes the nucleotide sequence shown in SEQ ID NO:106, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:201;
82)所述正义链包含如SEQ ID NO:107所示的核苷酸序列，所述反义链包含如SEQ ID NO:202所示的核苷酸序列；82) The sense strand includes the nucleotide sequence shown in SEQ ID NO:107, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:202;
83)所述正义链包含如SEQ ID NO:108所示的核苷酸序列，所述反义链包含如SEQ ID NO:203所示的核苷酸序列；83) The sense strand includes the nucleotide sequence shown in SEQ ID NO:108, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:203;
84)所述正义链包含如SEQ ID NO:109所示的核苷酸序列，所述反义链包含如SEQ ID NO:204所示的核苷酸序列；84) The sense strand includes the nucleotide sequence shown in SEQ ID NO:109, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:204;
85)所述正义链包含如SEQ ID NO:110所示的核苷酸序列，所述反义链包含如SEQ ID NO:205所示的核苷酸序列；85) The sense strand includes the nucleotide sequence shown in SEQ ID NO:110, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:205;
86)所述正义链包含如SEQ ID NO:111所示的核苷酸序列，所述反义链包含如SEQ ID NO:206所示的核苷酸序列；86) The sense strand includes the nucleotide sequence shown in SEQ ID NO:111, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:206;
87)所述正义链包含如SEQ ID NO:112所示的核苷酸序列，所述反义链包含如SEQ ID NO:207所示的核苷酸序列；87) The sense strand includes the nucleotide sequence shown in SEQ ID NO:112, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:207;
88)所述正义链包含如SEQ ID NO:113所示的核苷酸序列，所述反义链包含如SEQ ID NO:208所示的核苷酸序列；88) The sense strand includes the nucleotide sequence shown in SEQ ID NO:113, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:208;
89)所述正义链包含如SEQ ID NO:114所示的核苷酸序列，所述反义链包含如SEQ ID NO:209所示的核苷酸序列；89) The sense strand includes the nucleotide sequence shown in SEQ ID NO:114, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:209;
90)所述正义链包含如SEQ ID NO:115所示的核苷酸序列，所述反义链包含如SEQ ID NO:210所示的核苷酸序列；90) The sense strand includes the nucleotide sequence shown in SEQ ID NO:115, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:210;
91)所述正义链包含如SEQ ID NO:116所示的核苷酸序列，所述反义链包含如SEQ ID NO:211所示的核苷酸序列；91) The sense strand includes the nucleotide sequence shown in SEQ ID NO:116, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:211;
92)所述正义链包含如SEQ ID NO:117所示的核苷酸序列，所述反义链包含如SEQ ID NO:212所示的核苷酸序列；92) The sense strand includes the nucleotide sequence shown in SEQ ID NO:117, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:212;
93)所述正义链包含如SEQ ID NO:118所示的核苷酸序列，所述反义链包含如SEQ ID NO:213所示的核苷酸序列；93) The sense strand includes the nucleotide sequence shown in SEQ ID NO:118, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:213;
94)所述正义链包含如SEQ ID NO:119所示的核苷酸序列，所述反义链包含如SEQ ID NO:214所示的核苷酸序列；94) The sense strand includes the nucleotide sequence shown in SEQ ID NO:119, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:214;
95)所述正义链包含如SEQ ID NO:120所示的核苷酸序列，所述反义链包含如SEQ ID NO:215所示的核苷酸序列；95) The sense strand includes the nucleotide sequence shown in SEQ ID NO:120, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:215;
96)所述正义链包含如SEQ ID NO:258所示的核苷酸序列，所述反义链包含如SEQ ID NO:268所示的核苷酸序列；96) The sense strand includes the nucleotide sequence shown in SEQ ID NO:258, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:268;
97)所述正义链包含如SEQ ID NO:259所示的核苷酸序列，所述反义链包含如SEQ ID NO:269所示的核苷酸序列； 97) The sense strand includes the nucleotide sequence shown in SEQ ID NO:259, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:269;
98)所述正义链包含如SEQ ID NO:260所示的核苷酸序列，所述反义链包含如SEQ ID NO:270所示的核苷酸序列；98) The sense strand includes the nucleotide sequence shown in SEQ ID NO:260, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:270;
99)所述正义链包含如SEQ ID NO:261所示的核苷酸序列，所述反义链包含如SEQ ID NO:271所示的核苷酸序列；99) The sense strand includes the nucleotide sequence shown in SEQ ID NO:261, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:271;
100)所述正义链包含如SEQ ID NO:262所示的核苷酸序列，所述反义链包含如SEQ ID NO:272所示的核苷酸序列；100) The sense strand includes the nucleotide sequence shown in SEQ ID NO:262, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:272;
101)所述正义链包含如SEQ ID NO:263所示的核苷酸序列，所述反义链包含如SEQ ID NO:273所示的核苷酸序列；101) The sense strand includes the nucleotide sequence shown in SEQ ID NO:263, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:273;
102)所述正义链包含如SEQ ID NO:264所示的核苷酸序列，所述反义链包含如SEQ ID NO:274所示的核苷酸序列；102) The sense strand includes the nucleotide sequence shown in SEQ ID NO:264, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:274;
103)所述正义链包含如SEQ ID NO:265所示的核苷酸序列，所述反义链包含如SEQ ID NO:275所示的核苷酸序列；103) The sense strand includes the nucleotide sequence shown in SEQ ID NO:265, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:275;
104)所述正义链包含如SEQ ID NO:266所示的核苷酸序列，所述反义链包含如SEQ ID NO:276所示的核苷酸序列；104) The sense strand includes the nucleotide sequence shown in SEQ ID NO:266, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:276;
105)所述正义链包含如SEQ ID NO:267所示的核苷酸序列，所述反义链包含如SEQ ID NO:277所示的核苷酸序列；105) The sense strand includes the nucleotide sequence shown in SEQ ID NO:267, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:277;
137)所述正义链包含如SEQ ID NO:298所示的核苷酸序列，所述反义链包含如SEQ ID NO:321所示的核苷酸序列；137) The sense strand includes the nucleotide sequence shown in SEQ ID NO:298, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:321;
138)所述正义链包含如SEQ ID NO:299所示的核苷酸序列，所述反义链包含如SEQ ID NO:322所示的核苷酸序列；138) The sense strand includes the nucleotide sequence shown in SEQ ID NO:299, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:322;
139)所述正义链包含如SEQ ID NO:300所示的核苷酸序列，所述反义链包含如SEQ ID NO:323所示的核苷酸序列；139) The sense strand includes the nucleotide sequence shown in SEQ ID NO:300, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:323;
140)所述正义链包含如SEQ ID NO:301所示的核苷酸序列，所述反义链包含如SEQ ID NO:324所示的核苷酸序列；140) The sense strand includes the nucleotide sequence shown in SEQ ID NO:301, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:324;
141)所述正义链包含如SEQ ID NO:302所示的核苷酸序列，所述反义链包含如SEQ ID NO:325所示的核苷酸序列；141) The sense strand includes the nucleotide sequence shown in SEQ ID NO:302, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:325;
142)所述正义链包含如SEQ ID NO:303所示的核苷酸序列，所述反义链包含如SEQ ID NO:326所示的核苷酸序列；142) The sense strand includes the nucleotide sequence shown in SEQ ID NO:303, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:326;
143)所述正义链包含如SEQ ID NO:304所示的核苷酸序列，所述反义链包含如SEQ ID NO:327所示的核苷酸序列；143) The sense strand includes the nucleotide sequence shown in SEQ ID NO:304, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:327;
144)所述正义链包含如SEQ ID NO:305所示的核苷酸序列，所述反义链包含如SEQ ID NO:328所示的核苷酸序列；144) The sense strand includes the nucleotide sequence shown in SEQ ID NO:305, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:328;
145)所述正义链包含如SEQ ID NO:306所示的核苷酸序列，所述反义链包含如SEQ ID NO:329所示的核苷酸序列；145) The sense strand includes the nucleotide sequence shown in SEQ ID NO:306, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:329;
146)所述正义链包含如SEQ ID NO:307所示的核苷酸序列，所述反义链包含如SEQ ID NO:330所示的核苷酸序列；146) The sense strand includes the nucleotide sequence shown in SEQ ID NO:307, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:330;
147)所述正义链包含如SEQ ID NO:308所示的核苷酸序列，所述反义链包含如SEQ ID NO:331所示的核苷酸序列；147) The sense strand includes the nucleotide sequence shown in SEQ ID NO:308, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:331;
148)所述正义链包含如SEQ ID NO:309所示的核苷酸序列，所述反义链包含如SEQ ID NO:332所示的核苷酸序列；148) The sense strand includes the nucleotide sequence shown in SEQ ID NO:309, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:332;
149)所述正义链包含如SEQ ID NO:310所示的核苷酸序列，所述反义链包含如SEQ ID NO:333所示的核苷酸序列；149) The sense strand includes the nucleotide sequence shown in SEQ ID NO:310, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:333;
150)所述正义链包含如SEQ ID NO:311所示的核苷酸序列，所述反义链包含如SEQ ID NO:334所示的核苷酸序列；150) The sense strand includes the nucleotide sequence shown in SEQ ID NO:311, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:334;
151)所述正义链包含如SEQ ID NO:312所示的核苷酸序列，所述反义链包含如SEQ ID  NO:335所示的核苷酸序列；151) The sense strand comprises the nucleotide sequence shown in SEQ ID NO:312, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:312 The nucleotide sequence shown in NO:335;
152)所述正义链包含如SEQ ID NO:313所示的核苷酸序列，所述反义链包含如SEQ ID NO:336所示的核苷酸序列；152) The sense strand includes the nucleotide sequence shown in SEQ ID NO:313, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:336;
153)所述正义链包含如SEQ ID NO:314所示的核苷酸序列，所述反义链包含如SEQ ID NO:337所示的核苷酸序列；153) The sense strand includes the nucleotide sequence shown in SEQ ID NO:314, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:337;
154)所述正义链包含如SEQ ID NO:315所示的核苷酸序列，所述反义链包含如SEQ ID NO:338所示的核苷酸序列；154) The sense strand includes the nucleotide sequence shown in SEQ ID NO:315, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:338;
155)所述正义链包含如SEQ ID NO:316所示的核苷酸序列，所述反义链包含如SEQ ID NO:339所示的核苷酸序列；155) The sense strand includes the nucleotide sequence shown in SEQ ID NO:316, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:339;
156)所述正义链包含如SEQ ID NO:317所示的核苷酸序列，所述反义链包含如SEQ ID NO:340所示的核苷酸序列；156) The sense strand includes the nucleotide sequence shown in SEQ ID NO:317, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:340;
157)所述正义链包含如SEQ ID NO:318所示的核苷酸序列，所述反义链包含如SEQ ID NO:341所示的核苷酸序列；157) The sense strand includes the nucleotide sequence shown in SEQ ID NO:318, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:341;
158)所述正义链包含如SEQ ID NO:319所示的核苷酸序列，所述反义链包含如SEQ ID NO:342所示的核苷酸序列；158) The sense strand includes the nucleotide sequence shown in SEQ ID NO:319, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:342;
159)所述正义链包含如SEQ ID NO:320所示的核苷酸序列，所述反义链包含如SEQ ID NO:343所示的核苷酸序列；159) The sense strand includes the nucleotide sequence shown in SEQ ID NO:320, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:343;
201)所述正义链包含如SEQ ID NO:385所示的核苷酸序列，所述反义链包含如SEQ ID NO:387所示的核苷酸序列；201) The sense strand includes the nucleotide sequence shown in SEQ ID NO:385, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:387;
202)所述正义链包含如SEQ ID NO:386所示的核苷酸序列，所述反义链包含如SEQ ID NO:388所示的核苷酸序列。202) The sense strand includes the nucleotide sequence shown in SEQ ID NO:386, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:388.
[10].根据[1]-[9]任一项所述的双链核糖核酸，其中，所述正义链中每个核苷酸彼此独立地为修饰的核苷酸或未修饰的核苷酸，和/或，所述反义链中每个核苷酸彼此独立地为修饰的核苷酸或未修饰的核苷酸。[10]. The double-stranded ribonucleic acid according to any one of [1] to [9], wherein each nucleotide in the sense strand is independently a modified nucleotide or an unmodified nucleoside. acid, and/or, each nucleotide in the antisense strand is independently a modified nucleotide or an unmodified nucleotide.
[11].根据[1]-[10]任一项所述的双链核糖核酸，其中，所述正义链中任意相连的两个核苷酸由磷酸二酯键或硫代磷酸二酯键连接，和/或，所述反义链中任意相连的两个核苷酸由磷酸二酯键或硫代磷酸二酯键连接。[11]. The double-stranded ribonucleic acid according to any one of [1] to [10], wherein any two nucleotides connected in the sense strand are connected by a phosphodiester bond or a phosphorothioate diester bond. Connection, and/or, any two nucleotides connected in the antisense strand are connected by a phosphodiester bond or a phosphorothioate diester bond.
[12].根据[1]-[11]任一项所述的双链核糖核酸，其中，所述正义链的5’末端核苷酸连接5’磷酸基团或5’磷酸衍生基团，和/或，所述反义链的5’末端核苷酸连接5’磷酸基团或5’磷酸衍生基团。[12]. The double-stranded ribonucleic acid according to any one of [1] to [11], wherein the 5' terminal nucleotide of the sense strand is connected to a 5' phosphate group or a 5' phosphate derivative group, And/or, the 5' terminal nucleotide of the antisense strand is connected to a 5' phosphate group or a 5' phosphate derivative group.
[13].根据[1]-[12]任一项所述的双链核糖核酸，其中，所述双链核糖核酸为siRNA。[13]. The double-stranded ribonucleic acid according to any one of [1] to [12], wherein the double-stranded ribonucleic acid is siRNA.
[14].根据[1]-[13]任一项所述的双链核糖核酸，其中，所述双链核糖核酸为用于抑制ANGPTL3基因表达的siRNA。[14]. The double-stranded ribonucleic acid according to any one of [1] to [13], wherein the double-stranded ribonucleic acid is siRNA for inhibiting expression of ANGPTL3 gene.
[15].一种双链核糖核酸修饰物，其为如[1]-[14]任一项所述的双链核糖核酸的修饰物，所述双链核糖核酸修饰物包含如下至少一种的化学修饰：[15]. A double-stranded ribonucleic acid modification, which is a double-stranded ribonucleic acid modification as described in any one of [1]-[14], and the double-stranded ribonucleic acid modification includes at least one of the following Chemical modification:
(1)正义链中至少一个核苷酸的修饰，(1) Modification of at least one nucleotide in the sense strand,
(2)正义链中至少一个位置处的磷酸二酯键的修饰，(2) Modification of the phosphodiester bond at at least one position in the sense strand,
(3)反义链中至少一个核苷酸的修饰，(3) Modification of at least one nucleotide in the antisense strand,
(4)反义链中至少一个位置处的磷酸二酯键的修饰；(4) Modification of the phosphodiester bond at at least one position in the antisense strand;
任选地，所述双链核糖核酸的正义链中序列A的3’末端连接由1-2个核苷酸组成的序列D，优选由1-2个胸腺嘧啶脱氧核糖核苷酸组成的序列D；和/或，所述双链核糖核酸的反义链中序列B的3’末端连接由1-2个核苷酸组成的序列E，优选由1-2个胸腺嘧啶脱氧核糖核苷酸组成的序列E；和/或，所述双链核糖核酸的正义链中序列A的3’末端排除1-2个核苷酸后形成序列A’；Optionally, the 3' end of sequence A in the sense strand of the double-stranded ribonucleic acid is connected to sequence D consisting of 1-2 nucleotides, preferably a sequence consisting of 1-2 thymidine deoxyribonucleotides D; and/or, the 3' end of sequence B in the antisense strand of the double-stranded ribonucleic acid is connected to sequence E consisting of 1-2 nucleotides, preferably 1-2 thymidine deoxyribonucleotides Sequence E consisting of; and/or sequence A' is formed after excluding 1-2 nucleotides from the 3' end of sequence A in the sense strand of the double-stranded ribonucleic acid;
可选地，所述双链核糖核酸修饰物的正义链和反义链选自如下的序列组合： Alternatively, the sense strand and antisense strand of the double-stranded ribonucleic acid modification are selected from the following sequence combinations:
所述正义链的核苷酸序列为序列A所示的序列，所述反义链的核苷酸序列为序列B所示的序列；The nucleotide sequence of the sense strand is the sequence shown in sequence A, and the nucleotide sequence of the antisense strand is the sequence shown in sequence B;
或者，所述正义链的核苷酸序列为序列A所示的序列，所述反义链的核苷酸序列为序列B连接序列E所示的序列；Alternatively, the nucleotide sequence of the sense strand is the sequence shown in sequence A, and the nucleotide sequence of the antisense strand is the sequence shown in sequence B connected to sequence E;
或者，所述正义链的核苷酸序列为序列A连接序列D所示的序列，所述反义链的核苷酸序列为序列B所示的序列；Alternatively, the nucleotide sequence of the sense strand is the sequence shown in sequence A connected to sequence D, and the nucleotide sequence of the antisense strand is the sequence shown in sequence B;
或者，所述正义链的核苷酸序列为序列A连接序列D所示的序列，所述反义链的核苷酸序列为序列B连接序列E所示的序列；Alternatively, the nucleotide sequence of the sense strand is the sequence shown in sequence A connected to sequence D, and the nucleotide sequence of the antisense strand is the sequence shown in sequence B connected to sequence E;
或者，所述正义链的核苷酸序列为序列A’所示的序列，所述反义链的核苷酸序列为序列B所示的序列；Alternatively, the nucleotide sequence of the sense strand is the sequence shown in sequence A', and the nucleotide sequence of the antisense strand is the sequence shown in sequence B;
或者，所述正义链的核苷酸序列为序列A’所示的序列，所述反义链的核苷酸序列为序列B连接序列E所示的序列。Alternatively, the nucleotide sequence of the sense strand is the sequence represented by sequence A', and the nucleotide sequence of the antisense strand is the sequence represented by sequence B connected to sequence E.
[16].根据[15]所述的双链核糖核酸修饰物，其中，所述核苷酸的修饰选自2’-氟代修饰，2’-烷氧基修饰，2’-取代的烷氧基修饰，2’-烷基修饰，2’-取代的烷基修饰，2’-脱氧修饰，核苷酸衍生物修饰或其中任意两种以上的组合。[16]. The double-stranded ribonucleic acid modification according to [15], wherein the modification of the nucleotide is selected from the group consisting of 2'-fluoro modification, 2'-alkoxy modification, and 2'-substituted alkyl. Oxygen modification, 2'-alkyl modification, 2'-substituted alkyl modification, 2'-deoxy modification, nucleotide derivative modification or a combination of any two or more thereof.
[17].根据[15]或[16]所述的双链核糖核酸修饰物，其中，所述核苷酸的修饰选自2’-F修饰、2’-O-CH3修饰、2’-O-CH2-CH2-O-CH3修饰、2’-O-CH2-CH＝CH2修饰、2’-CH2-CH2-CH＝CH2修饰、2’-脱氧修饰，核苷酸衍生物修饰或其中任意两种以上的组合。[17]. The double-stranded ribonucleic acid modification according to [15] or [16], wherein the modification of the nucleotide is selected from the group consisting of 2'-F modification, 2'-O-CH 3 modification, and 2' -O-CH 2 -CH 2 -O-CH 3 modification, 2'-O-CH 2 -CH＝CH 2 modification, 2'-CH 2 -CH 2 -CH＝CH 2 modification, 2'-deoxy modification, Nucleotide derivative modification or a combination of any two or more thereof.
[18].根据[16]或[17]所述的双链核糖核酸修饰物，其中，所述核苷酸衍生物修饰中的核苷酸衍生物选自异核苷酸、LNA、ENA、cET、UNA或GNA。[18]. The double-stranded ribonucleic acid modification according to [16] or [17], wherein the nucleotide derivative in the nucleotide derivative modification is selected from the group consisting of isonucleotides, LNA, ENA, cET, UNA or GNA.
[19].根据[15]-[18]任一项所述的双链核糖核酸修饰物，其中，沿5’末端向3’末端方向，所述正义链中第7位、第9位、第10位和第11位的核糖核苷酸为2’-F修饰的核糖核苷酸，所述正义链中其余位置的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸。[19]. The double-stranded ribonucleic acid modification according to any one of [15] to [18], wherein, along the 5' end to the 3' end, the 7th, 9th, and 9th positions in the sense strand are The ribonucleotides at positions 10 and 11 are 2'-F modified ribonucleotides, and the remaining ribonucleotides in the sense strand are 2'-O-CH 3 modified ribonucleotides. .
[20].根据[15]-[19]任一项所述的双链核糖核酸修饰物，其中，沿5’末端向3’末端方向，所述正义链包含位于如下所示位置处的硫代磷酸二酯键：[20]. The double-stranded ribonucleic acid modification according to any one of [15] to [19], wherein, along the 5' end to the 3' end, the sense strand contains a sulfur molecule located at the position shown below Phosphodiester bond:
所述正义链5’末端起始的第1个核苷酸与第2个核苷酸之间；Between the first nucleotide and the second nucleotide starting from the 5' end of the sense strand;
所述正义链5’末端起始的第2个核苷酸与第3个核苷酸之间；Between the 2nd and 3rd nucleotide starting from the 5' end of the sense strand;
所述正义链3’末端起始的第1个核苷酸与第2个核苷酸之间；Between the first nucleotide and the second nucleotide starting from the 3' end of the sense strand;
所述正义链3’末端起始的第2个核苷酸与第3个核苷酸之间；Between the second and third nucleotides starting from the 3’ end of the sense strand;
或者，or,
所述正义链包含位于如下所示位置处的硫代磷酸二酯键：The sense strand contains phosphorothioate diester bonds at the positions shown below:
所述正义链5’末端起始的第1个核苷酸与第2个核苷酸之间；Between the first nucleotide and the second nucleotide starting from the 5' end of the sense strand;
所述正义链5’末端起始的第2个核苷酸与第3个核苷酸之间。Between the 2nd and 3rd nucleotide starting from the 5' end of the sense strand.
[21].根据[15]-[20]任一项所述的双链核糖核酸修饰物，其中，沿5’末端向3’末端方向，所述反义链中任意奇数位置处的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸，所述反义链中任意偶数位置处的核糖核苷酸为2’-F修饰的核糖核苷酸；[21]. The double-stranded ribonucleic acid modification according to any one of [15] to [20], wherein, along the 5' end to the 3' end, the ribonucleic acid at any odd position in the antisense strand The nucleotide is a 2'-O-CH 3 modified ribonucleotide, and the ribonucleotide at any even-numbered position in the antisense chain is a 2'-F modified ribonucleotide;
或者，沿5’末端向3’末端方向，所述反义链中第2位、第6位、第14位和第16位的核糖核苷酸为2’-F修饰的核糖核苷酸，所述反义链中其余位置的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸；Alternatively, along the direction from the 5' end to the 3' end, the ribonucleotides at positions 2, 6, 14 and 16 in the antisense strand are 2'-F modified ribonucleotides, The remaining ribonucleotides in the antisense strand are 2'-O-CH 3 modified ribonucleotides;
或者，沿5’末端向3’末端方向，所述反义链中第2位、第6位、第8位、第9位、第14位和第16位的核糖核苷酸为2’-F修饰的核糖核苷酸，所述反义链中其余位置的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸；Alternatively, along the direction from the 5' end to the 3' end, the ribonucleotides at positions 2, 6, 8, 9, 14 and 16 in the antisense strand are 2'- F-modified ribonucleotides, the remaining ribonucleotides in the antisense strand are 2'-O-CH 3 modified ribonucleotides;
或者，沿5’末端向3’末端方向，所述反义链中第2位、第14位和第16位的核糖核苷酸为2’-F修饰的核糖核苷酸，所述反义链中第6位的核糖核苷酸为核苷酸衍生物GNA修饰的核糖核苷酸，所述反义链中其余位置的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸； Alternatively, along the direction from the 5' end to the 3' end, the ribonucleotides at positions 2, 14 and 16 in the antisense strand are 2'-F modified ribonucleotides, and the antisense strand The ribonucleotide at position 6 in the chain is a ribonucleotide modified by the nucleotide derivative GNA, and the ribonucleotides at the remaining positions in the antisense chain are ribonucleotides modified by 2'-O-CH 3 acid;
或者，沿5’末端向3’末端方向，所述反义链中第2位、第6位、第14位和第16位的核糖核苷酸为2’-F修饰的核糖核苷酸，所述反义链中第7位的核糖核苷酸为核苷酸衍生物GNA修饰的核糖核苷酸，所述反义链中其余位置的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸。Alternatively, along the direction from the 5' end to the 3' end, the ribonucleotides at positions 2, 6, 14 and 16 in the antisense strand are 2'-F modified ribonucleotides, The ribonucleotide at position 7 in the antisense strand is a ribonucleotide modified by the nucleotide derivative GNA, and the ribonucleotides at the remaining positions in the antisense strand are modified with 2'-O-CH 3 of ribonucleotides.
[22].根据[15]-[21]任一项所述的双链核糖核酸修饰物，其中，沿5’末端向3’末端方向，所述反义链的5’末端的核苷酸连接5’磷酸基团或5’磷酸衍生基团。[22]. The double-stranded ribonucleic acid modification according to any one of [15] to [21], wherein, in the direction from the 5' end to the 3' end, the nucleotides at the 5' end of the antisense strand Attached 5' phosphate group or 5' phosphate derivative group.
[23].根据[15]-[22]任一项所述的双链核糖核酸修饰物，其中，所述反义链包含位于如下所示位置处的硫代磷酸二酯键：[23]. The double-stranded ribonucleic acid modification according to any one of [15] to [22], wherein the antisense strand contains a phosphorothioate diester bond located at the position shown below:
所述反义链5’末端起始的第1个核苷酸与第2个核苷酸之间；Between the first nucleotide and the second nucleotide starting from the 5' end of the antisense strand;
所述反义链5’末端起始的第2个核苷酸与第3个核苷酸之间；Between the 2nd and 3rd nucleotide starting from the 5' end of the antisense strand;
所述反义链3’末端起始的第1个核苷酸与第2个核苷酸之间；Between the first nucleotide and the second nucleotide starting from the 3' end of the antisense strand;
所述反义链3’末端起始的第2个核苷酸与第3个核苷酸之间。Between the 2nd and 3rd nucleotide starting from the 3' end of the antisense strand.
[24].根据[15]-[23]任一项所述的双链核糖核酸修饰物，其中，所述双链核糖核酸修饰物的正义链具有如(a1)-(a5)任一项所示的结构：[24]. The double-stranded ribonucleic acid modification according to any one of [15] to [23], wherein the sense strand of the double-stranded ribonucleic acid modification has any of (a 1 ) to (a 5 ) The structure shown in one item:
(a1)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-(s)-T-(s)-T-3’，(a 1 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3'，
(a2)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(a 2 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
(a3)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(a 3 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3',
(a4)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-3’，(a 4 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -3',
(a5)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-mN20-mN21-3’；(a 5 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -3';
其中，N1-N23彼此独立地选自碱基为A、U、C或G的核糖核苷酸，Wherein, N 1 -N 23 are independently selected from ribonucleotides whose bases are A, U, C or G,
大写字母T表示碱基为胸腺嘧啶的脱氧核糖核苷酸，The capital letter T stands for deoxyribonucleotide whose base is thymine.
小写字母m表示该字母m右侧相邻的一个核糖核苷酸为2’-O-CH3修饰的核糖核苷酸，The lowercase letter m indicates that the ribonucleotide adjacent to the right side of the letter m is a 2'-O-CH 3 modified ribonucleotide.
小写字母f表示该字母f左侧相邻的一个核糖核苷酸为2’-F修饰的核糖核苷酸，The lowercase letter f indicates that the ribonucleotide adjacent to the left side of the letter f is a 2’-F modified ribonucleotide.
-(s)-表示前后相邻的两个核苷酸以硫代磷酸二酯键连接。-(s)- indicates that two adjacent nucleotides are connected by a phosphorothioate diester bond.
[25].根据[15]-[24]任一项所述的双链核糖核酸修饰物，其中，所述双链核糖核酸修饰物的反义链具有如(b1)-(b15)任一项所示的结构：[25]. The double-stranded ribonucleic acid modification according to any one of [15] to [24], wherein the antisense strand of the double-stranded ribonucleic acid modification has a structure such as (b 1 )-(b 15 ) The structure shown in any one:
(b1)5’-P1mN1-(s)-N2f-(s)-mN3-N4f-mN5-N6f-mN7-N8f-mN9-N10f-mN11-N12f-mN13-N14f-mN15-N16f-mN17-N18f-mN19-(s)-T-(s)-T-3’，(b 1 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -N 4 f-mN 5 -N 6 f-mN 7 -N 8 f-mN 9 -N 10 f- mN 11 -N 12 f-mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -N 18 f-mN 19 -(s)-T-(s)-T-3',
(b2)5’-P1mN1-(s)-N2f-(s)-mN3-N4f-mN5-N6f-mN7-N8f-mN9-N10f-mN11-N12f-mN13-N14f-mN15-N16f-mN17-N18f-mN19-(s)-N20f-(s)-mN21-3’，(b 2 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -N 4 f-mN 5 -N 6 f-mN 7 -N 8 f-mN 9 -N 10 f- mN 11 -N 12 f-mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -N 18 f-mN 19 -(s)-N 20 f-(s)-mN 21 -3',
(b3)5’-P1mN1-(s)-N2f-(s)-mN3-N4f-mN5-N6f-mN7-N8f-mN9-N10f-mN11-N12f-mN13-N14f-mN15-N16f-mN17-N18f-mN19-N20f-mN21-(s)-N22f-(s)-mN23-3’,(b 3 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -N 4 f-mN 5 -N 6 f-mN 7 -N 8 f-mN 9 -N 10 f- mN 11 -N 12 f-mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -N 18 f-mN 19 -N 20 f-mN 21 -(s)-N 22 f-(s)- mN 23 -3',
(b4)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N1 4f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 4 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -mN 8 -mN 9 -mN 10 -mN 11 - mN 12 -mN 13 -N 1 4 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3'，
(b5)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N1 4f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 5 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -mN 8 -mN 9 -mN 10 -mN 11 - mN 12 -mN 13 -N 1 4 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
(b6)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N1 4f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(b 6 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -mN 8 -mN 9 -mN 10 -mN 11 - mN 12 -mN 13 -N 1 4 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3',
(b7)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-N8f-N9f-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 7 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -N 8 fN 9 f-mN 10 -mN 11 - mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3'，
(b8)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-N8f-N9f-mN10-mN11-mN12-mN13-N14f -mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 8 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -N 8 fN 9 f-mN 10 -mN 11 - mN 12 -mN 13 -N 14 f -mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
(b9)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-N8f-N9f-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(b 9 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -N 8 fN 9 f-mN 10 -mN 11 - mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3',
(b10)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-[GNA]N6-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 10 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -[GNA]N 6 -mN 7 -mN 8 -mN 9 -mN 10 -mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3',
(b11)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-[GNA]N6-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 11 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -[GNA]N 6 -mN 7 -mN 8 -mN 9 -mN 10 -mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
(b12)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-[GNA]N6-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(b 12 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -[GNA]N 6 -mN 7 -mN 8 -mN 9 -mN 10 -mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3' ,
(b13)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-[GNA]N7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 13 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-[GNA]N 7 -mN 8 -mN 9 -mN 10 - mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3',
(b14)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-[GNA]N7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 14 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-[GNA]N 7 -mN 8 -mN 9 -mN 10 - mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
(b15)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-[GNA]N7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’；(b 15 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-[GNA]N 7 -mN 8 -mN 9 -mN 10 - mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3 ';
其中，N1-N23彼此独立地选自碱基为A、U、C或G的核糖核苷酸，Wherein, N 1 -N 23 are independently selected from ribonucleotides whose bases are A, U, C or G,
大写字母T表示碱基为胸腺嘧啶的脱氧核糖核苷酸，The capital letter T stands for deoxyribonucleotide whose base is thymine.
小写字母m表示该字母m右侧相邻的一个核糖核苷酸为2’-O-CH3修饰的核糖核苷酸，The lowercase letter m indicates that the ribonucleotide adjacent to the right side of the letter m is a 2'-O-CH 3 modified ribonucleotide.
小写字母f表示该字母f左侧相邻的一个核糖核苷酸为2’-F修饰的核糖核苷酸，The lowercase letter f indicates that the ribonucleotide adjacent to the left side of the letter f is a 2’-F modified ribonucleotide.
P1表示该字母右侧相邻的一个核苷酸为5’-磷酸核苷酸，P1 indicates that the nucleotide adjacent to the right side of the letter is a 5’-phosphate nucleotide,
-(s)-表示前后相邻的两个核苷酸以硫代磷酸二酯键连接，-(s)- means that two adjacent nucleotides are connected by a phosphorothioate diester bond,
[GNA]表示其右侧相邻的一个核糖核苷酸为存在GNA修饰的核糖核苷酸。[GNA] indicates that the ribonucleotide adjacent to the right side is a ribonucleotide modified by GNA.
[26].根据[15]-[25]任一项所述的双链核糖核酸修饰物，其中，所述双链核糖核酸修饰物为siRNA修饰物。[26]. The double-stranded ribonucleic acid modified product according to any one of [15] to [25], wherein the double-stranded ribonucleic acid modified product is an siRNA modified product.
[27].根据[15]-[26]任一项所述的双链核糖核酸修饰物，其中，所述双链核糖核酸修饰物为用于抑制ANGPTL3基因表达的siRNA修饰物。[27]. The double-stranded ribonucleic acid modification according to any one of [15] to [26], wherein the double-stranded ribonucleic acid modification is an siRNA modification for inhibiting ANGPTL3 gene expression.
[28].根据[15]-[27]任一项所述的双链核糖核酸修饰物，其中，所述正义链与所述反义链选自如下组合：[28]. The double-stranded ribonucleic acid modification according to any one of [15] to [27], wherein the sense strand and the antisense strand are selected from the following combinations:
106)所述正义链包含如SEQ ID NO:216所示的核苷酸序列，所述反义链包含如SEQ ID NO:237所示的核苷酸序列；106) The sense strand includes the nucleotide sequence shown in SEQ ID NO:216, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:237;
107)所述正义链包含如SEQ ID NO:217所示的核苷酸序列，所述反义链包含如SEQ ID NO:238所示的核苷酸序列；107) The sense strand includes the nucleotide sequence shown in SEQ ID NO:217, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:238;
108)所述正义链包含如SEQ ID NO:218所示的核苷酸序列，所述反义链包含如SEQ ID NO:239所示的核苷酸序列；108) The sense strand includes the nucleotide sequence shown in SEQ ID NO:218, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:239;
109)所述正义链包含如SEQ ID NO:219所示的核苷酸序列，所述反义链包含如SEQ ID NO:240所示的核苷酸序列；109) The sense strand includes the nucleotide sequence shown in SEQ ID NO:219, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:240;
110)所述正义链包含如SEQ ID NO:220所示的核苷酸序列，所述反义链包含如SEQ ID NO:241所示的核苷酸序列；110) The sense strand includes the nucleotide sequence shown in SEQ ID NO:220, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:241;
111)所述正义链包含如SEQ ID NO:221所示的核苷酸序列，所述反义链包含如SEQ ID NO:242所示的核苷酸序列；111) The sense strand includes the nucleotide sequence shown in SEQ ID NO:221, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:242;
112)所述正义链包含如SEQ ID NO:222所示的核苷酸序列，所述反义链包含如SEQ ID NO:243所示的核苷酸序列；112) The sense strand includes the nucleotide sequence shown in SEQ ID NO:222, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:243;
113)所述正义链包含如SEQ ID NO:223所示的核苷酸序列，所述反义链包含如SEQ ID NO:244所示的核苷酸序列；113) The sense strand includes the nucleotide sequence shown in SEQ ID NO:223, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:244;
114)所述正义链包含如SEQ ID NO:224所示的核苷酸序列，所述反义链包含如SEQ ID  NO:245所示的核苷酸序列；114) The sense strand includes the nucleotide sequence shown in SEQ ID NO:224, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:224 The nucleotide sequence shown in NO:245;
115)所述正义链包含如SEQ ID NO:225所示的核苷酸序列，所述反义链包含如SEQ ID NO:246所示的核苷酸序列；115) The sense strand includes the nucleotide sequence shown in SEQ ID NO:225, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:246;
116)所述正义链包含如SEQ ID NO:226所示的核苷酸序列，所述反义链包含如SEQ ID NO:247所示的核苷酸序列；116) The sense strand includes the nucleotide sequence shown in SEQ ID NO:226, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:247;
117)所述正义链包含如SEQ ID NO:227所示的核苷酸序列，所述反义链包含如SEQ ID NO:248所示的核苷酸序列；117) The sense strand includes the nucleotide sequence shown in SEQ ID NO:227, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:248;
118)所述正义链包含如SEQ ID NO:228所示的核苷酸序列，所述反义链包含如SEQ ID NO:249所示的核苷酸序列；118) The sense strand includes the nucleotide sequence shown in SEQ ID NO:228, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:249;
119)所述正义链包含如SEQ ID NO:229所示的核苷酸序列，所述反义链包含如SEQ ID NO:250所示的核苷酸序列；119) The sense strand includes the nucleotide sequence shown in SEQ ID NO:229, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:250;
120)所述正义链包含如SEQ ID NO:230所示的核苷酸序列，所述反义链包含如SEQ ID NO:251所示的核苷酸序列；120) The sense strand includes the nucleotide sequence shown in SEQ ID NO:230, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:251;
121)所述正义链包含如SEQ ID NO:231所示的核苷酸序列，所述反义链包含如SEQ ID NO:252所示的核苷酸序列；121) The sense strand includes the nucleotide sequence shown in SEQ ID NO:231, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:252;
122)所述正义链包含如SEQ ID NO:232所示的核苷酸序列，所述反义链包含如SEQ ID NO:253所示的核苷酸序列；122) The sense strand includes the nucleotide sequence shown in SEQ ID NO:232, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:253;
123)所述正义链包含如SEQ ID NO:233所示的核苷酸序列，所述反义链包含如SEQ ID NO:254所示的核苷酸序列；123) The sense strand includes the nucleotide sequence shown in SEQ ID NO:233, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:254;
124)所述正义链包含如SEQ ID NO:234所示的核苷酸序列，所述反义链包含如SEQ ID NO:241所示的核苷酸序列；124) The sense strand includes the nucleotide sequence shown in SEQ ID NO:234, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:241;
125)所述正义链包含如SEQ ID NO:235所示的核苷酸序列，所述反义链包含如SEQ ID NO:247所示的核苷酸序列；125) The sense strand includes the nucleotide sequence shown in SEQ ID NO:235, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:247;
126)所述正义链包含如SEQ ID NO:236所示的核苷酸序列，所述反义链包含如SEQ ID NO:253所示的核苷酸序列；126) The sense strand includes the nucleotide sequence shown in SEQ ID NO:236, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:253;
127)所述正义链包含如SEQ ID NO:278所示的核苷酸序列，所述反义链包含如SEQ ID NO:288所示的核苷酸序列；127) The sense strand includes the nucleotide sequence shown in SEQ ID NO:278, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:288;
128)所述正义链包含如SEQ ID NO:279所示的核苷酸序列，所述反义链包含如SEQ ID NO:289所示的核苷酸序列；128) The sense strand includes the nucleotide sequence shown in SEQ ID NO:279, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:289;
129)所述正义链包含如SEQ ID NO:280所示的核苷酸序列，所述反义链包含如SEQ ID NO:290所示的核苷酸序列；129) The sense strand includes the nucleotide sequence shown in SEQ ID NO:280, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:290;
130)所述正义链包含如SEQ ID NO:281所示的核苷酸序列，所述反义链包含如SEQ ID NO:291所示的核苷酸序列；130) The sense strand includes the nucleotide sequence shown in SEQ ID NO:281, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:291;
131)所述正义链包含如SEQ ID NO:282所示的核苷酸序列，所述反义链包含如SEQ ID NO:292所示的核苷酸序列；131) The sense strand includes the nucleotide sequence shown in SEQ ID NO:282, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:292;
132)所述正义链包含如SEQ ID NO:283所示的核苷酸序列，所述反义链包含如SEQ ID NO:293所示的核苷酸序列；132) The sense strand includes the nucleotide sequence shown in SEQ ID NO:283, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:293;
133)所述正义链包含如SEQ ID NO:284所示的核苷酸序列，所述反义链包含如SEQ ID NO:294所示的核苷酸序列；133) The sense strand includes the nucleotide sequence shown in SEQ ID NO:284, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:294;
134)所述正义链包含如SEQ ID NO:285所示的核苷酸序列，所述反义链包含如SEQ ID NO:295所示的核苷酸序列；134) The sense strand includes the nucleotide sequence shown in SEQ ID NO:285, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:295;
135)所述正义链包含如SEQ ID NO:286所示的核苷酸序列，所述反义链包含如SEQ ID NO:296所示的核苷酸序列；135) The sense strand includes the nucleotide sequence shown in SEQ ID NO:286, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:296;
136)所述正义链包含如SEQ ID NO:287所示的核苷酸序列，所述反义链包含如SEQ ID NO:297所示的核苷酸序列； 136) The sense strand includes the nucleotide sequence shown in SEQ ID NO:287, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:297;
160)所述正义链包含如SEQ ID NO:344所示的核苷酸序列，所述反义链包含如SEQ ID NO:354所示的核苷酸序列；160) The sense strand includes the nucleotide sequence shown in SEQ ID NO:344, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:354;
161)所述正义链包含如SEQ ID NO:345所示的核苷酸序列，所述反义链包含如SEQ ID NO:355所示的核苷酸序列；161) The sense strand includes the nucleotide sequence shown in SEQ ID NO:345, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:355;
162)所述正义链包含如SEQ ID NO:346所示的核苷酸序列，所述反义链包含如SEQ ID NO:356所示的核苷酸序列；162) The sense strand includes the nucleotide sequence shown in SEQ ID NO:346, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:356;
163)所述正义链包含如SEQ ID NO:347所示的核苷酸序列，所述反义链包含如SEQ ID NO:357所示的核苷酸序列；163) The sense strand includes the nucleotide sequence shown in SEQ ID NO:347, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:357;
164)所述正义链包含如SEQ ID NO:348所示的核苷酸序列，所述反义链包含如SEQ ID NO:358所示的核苷酸序列；164) The sense strand includes the nucleotide sequence shown in SEQ ID NO:348, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:358;
165)所述正义链包含如SEQ ID NO:349所示的核苷酸序列，所述反义链包含如SEQ ID NO:359所示的核苷酸序列；165) The sense strand includes the nucleotide sequence shown in SEQ ID NO:349, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:359;
166)所述正义链包含如SEQ ID NO:350所示的核苷酸序列，所述反义链包含如SEQ ID NO:360所示的核苷酸序列；166) The sense strand includes the nucleotide sequence shown in SEQ ID NO:350, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:360;
167)所述正义链包含如SEQ ID NO:351所示的核苷酸序列，所述反义链包含如SEQ ID NO:361所示的核苷酸序列；167) The sense strand includes the nucleotide sequence shown in SEQ ID NO:351, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:361;
168)所述正义链包含如SEQ ID NO:231所示的核苷酸序列，所述反义链包含如SEQ ID NO:362所示的核苷酸序列；168) The sense strand includes the nucleotide sequence shown in SEQ ID NO:231, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:362;
169)所述正义链包含如SEQ ID NO:352所示的核苷酸序列，所述反义链包含如SEQ ID NO:362所示的核苷酸序列；169) The sense strand includes the nucleotide sequence shown in SEQ ID NO:352, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:362;
170)所述正义链包含如SEQ ID NO:231所示的核苷酸序列，所述反义链包含如SEQ ID NO:363所示的核苷酸序列；170) The sense strand includes the nucleotide sequence shown in SEQ ID NO:231, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:363;
171)所述正义链包含如SEQ ID NO:231所示的核苷酸序列，所述反义链包含如SEQ ID NO:364所示的核苷酸序列；171) The sense strand includes the nucleotide sequence shown in SEQ ID NO:231, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:364;
172)所述正义链包含如SEQ ID NO:231所示的核苷酸序列，所述反义链包含如SEQ ID NO:365所示的核苷酸序列；172) The sense strand includes the nucleotide sequence shown in SEQ ID NO:231, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:365;
173)所述正义链包含如SEQ ID NO:232所示的核苷酸序列，所述反义链包含如SEQ ID NO:366所示的核苷酸序列；173) The sense strand includes the nucleotide sequence shown in SEQ ID NO:232, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:366;
174)所述正义链包含如SEQ ID NO:236所示的核苷酸序列，所述反义链包含如SEQ ID NO:366所示的核苷酸序列；174) The sense strand includes the nucleotide sequence shown in SEQ ID NO:236, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:366;
175)所述正义链包含如SEQ ID NO:232所示的核苷酸序列，所述反义链包含如SEQ ID NO:367所示的核苷酸序列；175) The sense strand includes the nucleotide sequence shown in SEQ ID NO:232, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:367;
176)所述正义链包含如SEQ ID NO:232所示的核苷酸序列，所述反义链包含如SEQ ID NO:368所示的核苷酸序列；176) The sense strand includes the nucleotide sequence shown in SEQ ID NO:232, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:368;
177)所述正义链包含如SEQ ID NO:232所示的核苷酸序列，所述反义链包含如SEQ ID NO:369所示的核苷酸序列；177) The sense strand includes the nucleotide sequence shown in SEQ ID NO:232, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:369;
178)所述正义链包含如SEQ ID NO:233所示的核苷酸序列，所述反义链包含如SEQ ID NO:370所示的核苷酸序列；178) The sense strand includes the nucleotide sequence shown in SEQ ID NO:233, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:370;
179)所述正义链包含如SEQ ID NO:353所示的核苷酸序列，所述反义链包含如SEQ ID NO:370所示的核苷酸序列；179) The sense strand includes the nucleotide sequence shown in SEQ ID NO:353, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:370;
180)所述正义链包含如SEQ ID NO:233所示的核苷酸序列，所述反义链包含如SEQ ID NO:371所示的核苷酸序列；180) The sense strand includes the nucleotide sequence shown in SEQ ID NO:233, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:371;
181)所述正义链包含如SEQ ID NO:233所示的核苷酸序列，所述反义链包含如SEQ ID NO:372所示的核苷酸序列；181) The sense strand includes the nucleotide sequence shown in SEQ ID NO:233, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:372;
182)所述正义链包含如SEQ ID NO:233所示的核苷酸序列，所述反义链包含如SEQ ID  NO:373所示的核苷酸序列；182) The sense strand comprises the nucleotide sequence shown in SEQ ID NO:233, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:233 The nucleotide sequence shown in NO:373;
183)所述正义链包含如SEQ ID NO:389所示的核苷酸序列，所述反义链包含如SEQ ID NO:381所示的核苷酸序列；183) The sense strand includes the nucleotide sequence shown in SEQ ID NO:389, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:381;
184)所述正义链包含如SEQ ID NO:389所示的核苷酸序列，所述反义链包含如SEQ ID NO:382所示的核苷酸序列；184) The sense strand includes the nucleotide sequence shown in SEQ ID NO:389, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:382;
185)所述正义链包含如SEQ ID NO:389所示的核苷酸序列，所述反义链包含如SEQ ID NO:383所示的核苷酸序列；185) The sense strand includes the nucleotide sequence shown in SEQ ID NO:389, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:383;
186)所述正义链包含如SEQ ID NO:389所示的核苷酸序列，所述反义链包含如SEQ ID NO:384所示的核苷酸序列；186) The sense strand includes the nucleotide sequence shown in SEQ ID NO:389, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:384;
187)所述正义链包含如SEQ ID NO:390所示的核苷酸序列，所述反义链包含如SEQ ID NO:394所示的核苷酸序列；187) The sense strand includes the nucleotide sequence shown in SEQ ID NO:390, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:394;
188)所述正义链包含如SEQ ID NO:390所示的核苷酸序列，所述反义链包含如SEQ ID NO:395所示的核苷酸序列；188) The sense strand includes the nucleotide sequence shown in SEQ ID NO:390, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:395;
189)所述正义链包含如SEQ ID NO:391所示的核苷酸序列，所述反义链包含如SEQ ID NO:395所示的核苷酸序列；189) The sense strand includes the nucleotide sequence shown in SEQ ID NO:391, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:395;
190)所述正义链包含如SEQ ID NO:390所示的核苷酸序列，所述反义链包含如SEQ ID NO:396所示的核苷酸序列；190) The sense strand includes the nucleotide sequence shown in SEQ ID NO:390, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:396;
191)所述正义链包含如SEQ ID NO:390所示的核苷酸序列，所述反义链包含如SEQ ID NO:397所示的核苷酸序列；191) The sense strand includes the nucleotide sequence shown in SEQ ID NO:390, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:397;
192)所述正义链包含如SEQ ID NO:392所示的核苷酸序列，所述反义链包含如SEQ ID NO:398所示的核苷酸序列；192) The sense strand includes the nucleotide sequence shown in SEQ ID NO:392, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:398;
193)所述正义链包含如SEQ ID NO:392所示的核苷酸序列，所述反义链包含如SEQ ID NO:399所示的核苷酸序列；193) The sense strand includes the nucleotide sequence shown in SEQ ID NO:392, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:399;
194)所述正义链包含如SEQ ID NO:393所示的核苷酸序列，所述反义链包含如SEQ ID NO:399所示的核苷酸序列；194) The sense strand includes the nucleotide sequence shown in SEQ ID NO:393, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:399;
195)所述正义链包含如SEQ ID NO:392所示的核苷酸序列，所述反义链包含如SEQ ID NO:400所示的核苷酸序列；195) The sense strand includes the nucleotide sequence shown in SEQ ID NO:392, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:400;
196)所述正义链包含如SEQ ID NO:392所示的核苷酸序列，所述反义链包含如SEQ ID NO:401所示的核苷酸序列；196) The sense strand includes the nucleotide sequence shown in SEQ ID NO:392, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:401;
197)所述正义链包含如SEQ ID NO:226所示的核苷酸序列，所述反义链包含如SEQ ID NO:377所示的核苷酸序列；197) The sense strand includes the nucleotide sequence shown in SEQ ID NO:226, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:377;
198)所述正义链包含如SEQ ID NO:226所示的核苷酸序列，所述反义链包含如SEQ ID NO:378所示的核苷酸序列；198) The sense strand includes the nucleotide sequence shown in SEQ ID NO:226, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:378;
199)所述正义链包含如SEQ ID NO:226所示的核苷酸序列，所述反义链包含如SEQ ID NO:402所示的核苷酸序列；199) The sense strand includes the nucleotide sequence shown in SEQ ID NO:226, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:402;
200)所述正义链包含如SEQ ID NO:226所示的核苷酸序列，所述反义链包含如SEQ ID NO:403所示的核苷酸序列。200) The sense strand includes the nucleotide sequence shown in SEQ ID NO:226, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:403.
[29].一种双链核糖核酸缀合物，其中，所述双链核糖核酸缀合物包括如[1]-[14]任一项所述的双链核糖核酸，或如[15]-[28]任一项所述的双链核糖核酸修饰物；以及，缀合连接于所述双链核糖核酸或所述双链核糖核酸修饰物的缀合基团。[29]. A double-stranded ribonucleic acid conjugate, wherein the double-stranded ribonucleic acid conjugate includes the double-stranded ribonucleic acid as described in any one of [1]-[14], or as [15] - The double-stranded ribonucleic acid modification according to any one of [28]; and, a conjugation group connected to the double-stranded ribonucleic acid or the double-stranded ribonucleic acid modification.
[30].根据[29]所述的双链核糖核酸缀合物，其中，所述缀合基团具有如下所示结构：
[30]. The double-stranded ribonucleic acid conjugate according to [29], wherein the conjugation group has the following structure:
[31].根据[29]或[30]所述的双链核糖核酸缀合物，其中，所述缀合基团连接于正义链的3’末端。[31]. The double-stranded ribonucleic acid conjugate according to [29] or [30], wherein the conjugation group is connected to the 3’ end of the sense strand.
[32].根据[31]所述的双链核糖核酸缀合物，其中，所述缀合基团通过磷酸二酯键与正义链的3’末端缀合连接；[32]. The double-stranded ribonucleic acid conjugate according to [31], wherein the conjugation group is conjugated to the 3’ end of the sense strand through a phosphodiester bond;
优选地，所述双链核糖核酸缀合物的正义链与反义链互补形成所述双链核糖核酸缀合物的双链区，且所述正义链的3’末端形成平末端，所述反义链的3’末端具有1-2个延伸出所述双链区的突出的核苷酸；Preferably, the sense strand and the antisense strand of the double-stranded ribonucleic acid conjugate are complementary to form a double-stranded region of the double-stranded ribonucleic acid conjugate, and the 3' end of the sense strand forms a blunt end, and the The 3' end of the antisense strand has 1-2 protruding nucleotides extending out of the double-stranded region;
或者，or,
所述双链核糖核酸缀合物的正义链与反义链互补形成所述双链核糖核酸缀合物的双链区，且所述正义链的3’末端形成平末端，所述反义链的3’末端形成平末端。The sense strand and the antisense strand of the double-stranded ribonucleic acid conjugate are complementary to form the double-stranded region of the double-stranded ribonucleic acid conjugate, and the 3' end of the sense strand forms a blunt end, and the antisense strand The 3' end forms a blunt end.
[33].根据[29]-[32]任一项所述的双链核糖核酸缀合物，其中，所述双链核糖核酸缀合物具有如下所示结构：
[33]. The double-stranded ribonucleic acid conjugate according to any one of [29]-[32], wherein the double-stranded ribonucleic acid conjugate has the following structure:
其中，双螺旋结构为双链核糖核酸或双链核糖核酸修饰物。Among them, the double helix structure is double-stranded ribonucleic acid or double-stranded ribonucleic acid modification.
[34].根据[29]-[33]任一项所述的双链核糖核酸缀合物，其中，所述双链核糖核酸缀合物为siRNA缀合物。[34]. The double-stranded ribonucleic acid conjugate according to any one of [29] to [33], wherein the double-stranded ribonucleic acid conjugate is an siRNA conjugate.
[35].根据[29]-[34]任一项所述的双链核糖核酸缀合物，其中，所述双链核糖核酸缀合物是用于抑制ANGPTL3基因表达的siRNA缀合物。[35]. The double-stranded ribonucleic acid conjugate according to any one of [29] to [34], wherein the double-stranded ribonucleic acid conjugate is an siRNA conjugate for inhibiting ANGPTL3 gene expression.
[36].根据[29]-[35]任一项所述的双链核糖核酸缀合物，其中，所述双链核糖核酸缀合物由表1所示的任意一种siRNA与缀合基团连接形成，或者，所述双链核糖核酸缀合物由表1-1所示的任意一种siRNA与缀合基团连接形成，或者，所述双链核糖核酸缀合物由表2所示的任意一种siRNA修饰物与缀合基团连接形成；[36]. The double-stranded ribonucleic acid conjugate according to any one of [29] to [35], wherein the double-stranded ribonucleic acid conjugate is composed of any siRNA shown in Table 1 conjugated to The double-stranded ribonucleic acid conjugate is formed by connecting any siRNA shown in Table 1-1 and the conjugating group, or the double-stranded ribonucleic acid conjugate is formed by connecting the conjugation group with the double-stranded ribonucleic acid conjugate. Any of the siRNA modifications shown is connected to a conjugating group;
优选地，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:255所示的核苷酸序列，所述反义链包含如SEQ ID NO:241所示的核苷酸序列；Preferably, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:255, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:241 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:256所示的核苷酸序列，所述反义链包含如SEQ ID NO:247所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:256, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:247 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:257所示的核苷酸序列，所述反义链包含如SEQ ID NO:253所示的核苷酸序列； Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:257, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:253. The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:256所示的核苷酸序列，所述反义链包含如SEQ ID NO:377所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 256, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 377 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:256所示的核苷酸序列，所述反义链包含如SEQ ID NO:378所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 256, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 378 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:256所示的核苷酸序列，所述反义链包含如SEQ ID NO:379所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 256, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 379 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:256所示的核苷酸序列，所述反义链包含如SEQ ID NO:380所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 256, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 380 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:257所示的核苷酸序列，所述反义链包含如SEQ ID NO:366所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 257, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 366 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:257所示的核苷酸序列，所述反义链包含如SEQ ID NO:367所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 257, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 367 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:257所示的核苷酸序列，所述反义链包含如SEQ ID NO:368所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 257, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 368 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:257所示的核苷酸序列，所述反义链包含如SEQ ID NO:369所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 257, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 369 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:374所示的核苷酸序列，所述反义链包含如SEQ ID NO:362所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:374, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:362 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:374所示的核苷酸序列，所述反义链包含如SEQ ID NO:363所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:374, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:363 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:374所示的核苷酸序列，所述反义链包含如SEQ ID NO:364所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:374, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:364 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:374所示的核苷酸序列，所述反义链包含如SEQ ID NO:365所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:374, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:365 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:375所示的核苷酸序列，所述反义链包含如SEQ ID NO:370所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:375, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:370 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:375所示的核苷酸序列，所述反义链包含如SEQ ID NO:371所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:375, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:371 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:375所示的核苷酸序列，所述反义链包含如SEQ ID NO:372所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:375, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:372 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:375所示的核苷酸序列，所述反义链包含如SEQ ID NO:373所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:375, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:373 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:376所示的核苷酸序列，所述反义链包含如SEQ ID NO:381所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:376, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:381 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:376所示的核苷酸序列，所述反义链包含如SEQ ID NO:382所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:376, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:382 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:376所示的核苷酸序列，所述反义链包含如SEQ ID NO:383所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:376, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:383 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:376所示的核苷酸序列，所述反义链包含如SEQ ID NO:384所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:376, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:384 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:374所示的核苷酸序列，所述反义链包含如SEQ ID NO:252所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 374, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 252 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:375所示的核苷酸序列，所述反义链包含如SEQ ID NO:254所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 375, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 254 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:404 所示的核苷酸序列，所述反义链包含如SEQ ID NO:394所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes SEQ ID NO: 404 The nucleotide sequence shown, the antisense strand includes the nucleotide sequence shown in SEQ ID NO:394;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:404所示的核苷酸序列，所述反义链包含如SEQ ID NO:395所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 404, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 395 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:404所示的核苷酸序列，所述反义链包含如SEQ ID NO:396所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 404, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 396 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:404所示的核苷酸序列，所述反义链包含如SEQ ID NO:397所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 404, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 397 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:405所示的核苷酸序列，所述反义链包含如SEQ ID NO:398所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 405, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 398 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:405所示的核苷酸序列，所述反义链包含如SEQ ID NO:399所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 405, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 399 The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:405所示的核苷酸序列，所述反义链包含如SEQ ID NO:400所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:405, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:400. The nucleotide sequence shown;
或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:405所示的核苷酸序列，所述反义链包含如SEQ ID NO:401所示的核苷酸序列。Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:405, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:401 The nucleotide sequence shown.
[37].一种药物组合物，其中，所述药物组合物包括如下至少一项：如[1]-[14]任一项所述的双链核糖核酸，如[15]-[28]任一项所述的双链核糖核酸修饰物，如[29]-[36]任一项所述的双链核糖核酸缀合物。[37]. A pharmaceutical composition, wherein the pharmaceutical composition includes at least one of the following: double-stranded ribonucleic acid as described in any one of [1]-[14], such as [15]-[28] The double-stranded ribonucleic acid modification described in any one of [29] to [36] is the double-stranded ribonucleic acid conjugate described in any one of [29] to [36].
[38].根据[37]所述的药物组合物，其中，所述药物组合物还包括一种或多种药学上可接收的载体。[38]. The pharmaceutical composition according to [37], wherein the pharmaceutical composition further includes one or more pharmaceutically acceptable carriers.
[39].根据[1]-[14]任一项所述的双链核糖核酸，根据[15]-[28]任一项所述的双链核糖核酸修饰物，根据[29]-[36]任一项所述的双链核糖核酸缀合物，或根据[37]-[38]任一项所述的药物组合物在如下至少一项中的用途：[39]. The double-stranded ribonucleic acid according to any one of [1]-[14], the double-stranded ribonucleic acid modification according to any one of [15]-[28], according to [29]-[ 36] The double-stranded ribonucleic acid conjugate according to any one of them, or the use of the pharmaceutical composition according to any one of [37]-[38] in at least one of the following:
(1)抑制ANGPTL3基因表达，或制备用于抑制ANGPTL3基因表达的药物；(1) Inhibit ANGPTL3 gene expression, or prepare drugs for inhibiting ANGPTL3 gene expression;
(2)用于预防或治疗与ANGPTL3基因异常表达相关的疾病，或制备用于预防或治疗与ANGPTL3基因异常表达相关的疾病的药物；(2) For preventing or treating diseases related to abnormal expression of ANGPTL3 gene, or preparing drugs for preventing or treating diseases related to abnormal expression of ANGPTL3 gene;
(3)用于治疗患有将受益于ANGPTL3基因表达降低的疾病的受试者，或制备用于治疗患有将受益于ANGPTL3基因表达降低的疾病的受试者的药物。(3) For use in treating subjects suffering from a disease that would benefit from a reduction in ANGPTL3 gene expression, or to prepare a medicament for treating subjects suffering from a disease that would benefit from a reduction in ANGPTL3 gene expression.
[40].根据[39]所述的用途，其中，所述与ANGPTL3基因异常表达相关的疾病选自与脂代谢相关的疾病；[40]. The use according to [39], wherein the diseases related to abnormal expression of ANGPTL3 gene are selected from diseases related to lipid metabolism;
可选地，所述与脂代谢相关的疾病选自如下疾病组成的组：Optionally, the disease related to lipid metabolism is selected from the group consisting of the following diseases:
乳糜微粒血症综合征、2型糖尿病、家族性部分脂肪营养不良、高胆固醇血症、家族性高胆固醇血症、高甘油三酯血症、非酒精性脂肪肝、动脉粥样硬化、高甘油三酯血症、膝盖损伤及骨关节炎、血脂异常。Chylomicronemia syndrome, type 2 diabetes, familial partial lipodystrophy, hypercholesterolemia, familial hypercholesterolemia, hypertriglyceridemia, non-alcoholic fatty liver disease, atherosclerosis, hyperglycemia Triesteremia, knee injury and osteoarthritis, dyslipidemia.
[41].一种用于抑制细胞内ANGPTL3基因表达的方法，其中，所述方法包括将所述细胞与根据[1]-[14]任一项所述的双链核糖核酸，根据[15]-[28]任一项所述的双链核糖核酸修饰物，根据[29]-[36]任一项所述的双链核糖核酸缀合物，或根据[37]-[38]任一项所述的药物组合物接触。[41]. A method for inhibiting ANGPTL3 gene expression in cells, wherein the method includes combining the cell with the double-stranded ribonucleic acid according to any one of [1]-[14], according to [15 ]-[28] The double-stranded ribonucleic acid modification according to any one of [29]-[36], or the double-stranded ribonucleic acid conjugate according to any one of [29]-[36], or according to any one of [37]-[38] Contact with the pharmaceutical composition described in one item.
[42].根据[41]所述的方法，其中，所述细胞为体内细胞或体外细胞。[42]. The method according to [41], wherein the cells are in vivo cells or in vitro cells.
[43].根据[41]或[42]所述的方法，其中，所述细胞在受试者体内。[43]. The method according to [41] or [42], wherein the cells are in the subject.
[44].根据[43]所述的方法，其中，所述受试者为哺乳动物，优选为人。[44]. The method according to [43], wherein the subject is a mammal, preferably a human.
[45].根据[43]或[44]所述的方法，其中，所述受试者具有如下至少一种特性：[45]. The method according to [43] or [44], wherein the subject has at least one of the following characteristics:
体内ANGPTL3基因异常表达，更具体地为ANGPTL3基因异常高表达；Abnormal expression of ANGPTL3 gene in vivo, more specifically abnormal high expression of ANGPTL3 gene;
患有与ANGPTL3基因异常表达相关的疾病；Suffering from diseases related to abnormal expression of ANGPTL3 gene;
患有将受益于ANGPTL3基因表达降低的疾病。 People with diseases that would benefit from reduced ANGPTL3 gene expression.
发明的效果Effect of the invention
在一些实施方案中，本公开提供的双链核糖核酸，能够在细胞内结合形成RNA诱导沉默复合物(RISC)，切割ANGPTL3基因转录的mRNA，高效、特异地抑制ANGPTL3基因的表达，用于治疗包括脂质代谢异常在内的ANGPTL3相关疾病。In some embodiments, the double-stranded ribonucleic acid provided by the present disclosure can combine in cells to form an RNA-induced silencing complex (RISC), cleave the mRNA transcribed by the ANGPTL3 gene, efficiently and specifically inhibit the expression of the ANGPTL3 gene, and is used for treatment ANGPTL3-related diseases including abnormalities in lipid metabolism.
进一步的，本公开中双链核糖核酸为siRNA，siRNA靶向结合并降解ANGPTL3基因的转录产物mRNA，发挥RNA干扰的作用，抑制ANGPTL3基因的蛋白表达，是一种抑制率高且特异性好的ANGPTL3抑制剂。Furthermore, in the present disclosure, the double-stranded ribonucleic acid is siRNA. siRNA targets and degrades the transcript product of the ANGPTL3 gene, mRNA, exerts the effect of RNA interference, inhibits the protein expression of the ANGPTL3 gene, and is a method with high inhibition rate and good specificity. ANGPTL3 inhibitors.
在一些实施方案中，本公开对双链核糖核酸进行修饰，得到双链核糖核酸修饰物，双链核糖核酸修饰物的稳定性高，适合体内疾病治疗中的应用。In some embodiments, the present disclosure modifies double-stranded ribonucleic acid to obtain a double-stranded ribonucleic acid modified product. The double-stranded ribonucleic acid modified product has high stability and is suitable for use in in vivo disease treatment.
进一步的，双链核糖核酸修饰物为siRNA修饰物，具有高的稳定性和较好的抑制活性。Furthermore, the double-stranded ribonucleic acid modification is a siRNA modification, which has high stability and good inhibitory activity.
在一些实施方案中，本公开在双链核糖核酸、双链核糖核酸修饰物上连接缀合基团得到双链核糖核酸或双链核糖核酸修饰物的缀合物，能够用于向组织、细胞中高效靶向递送，降低双链核糖核酸或双链核糖核酸修饰物对非靶向的正常组织、细胞的影响，提高其在临床疾病治疗中的安全性。In some embodiments, the present disclosure connects a conjugation group to double-stranded ribonucleic acid or double-stranded ribonucleic acid modifications to obtain a conjugate of double-stranded ribonucleic acid or double-stranded ribonucleic acid modifications, which can be used to deliver tissues and cells. Medium and high-efficiency targeted delivery reduces the impact of double-stranded ribonucleic acid or double-stranded ribonucleic acid modifications on non-targeted normal tissues and cells, and improves its safety in clinical disease treatment.
进一步的，双链核糖核酸缀合物为siRNA缀合物，在保持siRNA抑制活性、稳定性的同时，兼具器官或组织靶向性，可降低对其他组织或器官的影响以及减少siRNA分子使用量，可达到减轻毒性和降低成本的目的。Furthermore, the double-stranded ribonucleic acid conjugate is an siRNA conjugate. While maintaining the inhibitory activity and stability of siRNA, it also has organ or tissue targeting, which can reduce the impact on other tissues or organs and reduce the use of siRNA molecules. quantity, can achieve the purpose of reducing toxicity and reducing costs.
进一步的，本公开中的缀合基团为式I所示结构的基团(GalNAc)，GalNAc可用于向肝脏细胞、组织内的靶向递送，用于高效抑制肝脏内ANGPTL3基因的表达。Furthermore, the conjugation group in the present disclosure is a group (GalNAc) with the structure shown in Formula I. GalNAc can be used for targeted delivery to liver cells and tissues, and can be used to effectively inhibit the expression of ANGPTL3 gene in the liver.
具体实施方式Detailed ways
定义definition
除非有相反陈述，否则在本发明中所使用的术语具有下述含义。Unless stated to the contrary, the terms used in the present invention have the following meanings.
在本发明的权利要求和/或说明书中，词语“一(a)”或“一(an)”或“一(the)”可以指“一个”，但也可以指“一个或多个”、“至少一个”以及“一个或多于一个”。In the claims and/or description of the present invention, the word "a" or "an" or "the" may refer to "one", but may also refer to "one or more", "At least one" and "One or more than one".
如在权利要求和说明书中所使用的，词语“包含”、“具有”、“包括”或“含有”是指包括在内的或开放式的，并不排除额外的、未引述的元件或方法步骤。As used in the claims and description, the words "comprises," "having," "including," or "containing" mean inclusive or open-ended and do not exclude additional, unrecited elements or methods. step.
在整个申请文件中，术语“约”表示：一个值包括测定该值所使用的装置或方法的误差的标准偏差。用以界定本发明的数值范围与参数皆是约略的数值，此处已尽可能精确地呈现具体实施例中的相关数值。然而，任何数值本质上不可避免地含有因前述测试方法或装置所致的标准偏差。因此，除非另有明确的说明，应当理解本公开所用的所有范围、数量、数值与百分比均经过“约”的修饰。在此处，“约”通常是指实际数值在一特定数值或范围的正负10％、5％、1％或0.5％之内。Throughout this application, the term "about" means that a value includes the standard deviation of the error of the device or method used to determine the value. The numerical ranges and parameters used to define the present invention are approximate values, and the relevant values in the specific embodiments are presented as accurately as possible. Any numerical value, however, inherently contains standard deviations resulting from any such testing method or apparatus. Therefore, unless otherwise expressly stated, it should be understood that all ranges, quantities, values and percentages used in this disclosure are modified by "about". As used herein, "about" generally means that the actual value is within plus or minus 10%, 5%, 1% or 0.5% of a particular value or range.
本公开上下文中使用的术语“ANGPTL3”、“ANL3”、“angiopoietin like 3”，是指熟知的基因和多肽，也在本领域已知为：ANG-5、FHBL2、ANGPT5。ANGPTL3基因、ANGPTL3mRNA序列是例如使用以下容易获得的：基因库(GenBank)、数据库(UniProt)、人类孟德尔遗传在线(OMIM)等。The terms "ANGPTL3", "ANL3", "angiopoietin like 3", as used in the context of this disclosure, refer to well-known genes and polypeptides also known in the art: ANG-5, FHBL2, ANGPT5. ANGPTL3 gene and ANGPTL3 mRNA sequences are easily obtained using, for example, GenBank (GenBank), database (UniProt), Online Mendelian Inheritance in Man (OMIM), etc.
术语“ANGPTL3基因”，可以是野生型ANGPTL3基因，或存在序列变异的ANGPTL3基因突变体。在ANGPTL3基因中的许多序列变异已经被鉴别并且可以发现在例如NCBIdbSNP和UniProt(参见，例如，ncbi.nlm.nih.gov/snp)中。The term "ANGPTL3 gene" can be a wild-type ANGPTL3 gene or an ANGPTL3 gene mutant with sequence variation. Many sequence variations in the ANGPTL3 gene have been identified and can be found, for example, in NCBIdbSNP and UniProt (see, eg, ncbi.nlm.nih.gov/snp).
术语“多肽”、“蛋白”可互换地指通过共价键(例如肽键)相互连接的一串至少两个氨基酸残基，可以是重组多肽、天然多肽或合成多肽。多肽可以是线形或分支的，它可以包含修饰的氨基酸，并且它可以由非氨基酸隔断。该术语也包括已经被修饰(例如， 二硫键形成、糖基化、脂质化、乙酰化、磷酸化或任何其他操作，如以标记组分缀合)的氨基酸聚合物。The terms "polypeptide" and "protein" interchangeably refer to a sequence of at least two amino acid residues linked to each other by covalent bonds (eg, peptide bonds), and may be recombinant polypeptides, natural polypeptides, or synthetic polypeptides. A polypeptide can be linear or branched, it can contain modified amino acids, and it can be interrupted by non-amino acids. The term also includes being modified (e.g., disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation or any other manipulation, such as conjugation with a labeling component) of the amino acid polymer.
本公开上下文中使用的术语“靶标序列”是指在靶基因转录期间形成的mRNA分子的核苷酸序列的连续部分，包括作为对初级转录产物进行RNA加工的产物的mRNA。The term "target sequence" as used in the context of this disclosure refers to the contiguous portion of the nucleotide sequence of the mRNA molecule formed during the transcription of the target gene, including the mRNA as a product of RNA processing of the primary transcript.
在一些实施方案中，靶标序列是不少于16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、50、80、100、150、200、300、400、500、600或700个连续连接核苷组成的核苷酸序列。示例性的，靶标序列是19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、38、53、54、65、125、69、97、80、128、164或190个连续连接核苷组成的核苷酸序列。在一些可选的实施方案中，靶标序列中可以包含另一段较短的靶标序列。在一些实施方案中，靶标序列中可以包含一个或多个较短的靶标序列。应当认为，被包含于同一段靶标序列中的两个以上的较短靶标序列之间具有相同的特征。In some embodiments, the target sequence is no less than 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 50, 80, 100, 150, A nucleotide sequence consisting of 200, 300, 400, 500, 600 or 700 consecutively linked nucleosides. Exemplarily, the target sequences are 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 38, 53, 54, 65, 125 , 69, 97, 80, 128, 164 or 190 nucleotide sequences composed of consecutively connected nucleosides. In some alternative embodiments, another shorter target sequence may be included in the target sequence. In some embodiments, one or more shorter target sequences may be included in the target sequence. It should be considered that two or more shorter target sequences included in the same target sequence have the same characteristics.
在一些实施方案中，靶基因为ANGPTL3基因。在一些实施方案中，序列的靶部分将会是至少足够地长，以在ANGPTL3基因的转录期间形成的mRNA分子的核苷酸序列部分处或其附近充当iRNA指导的切割的底物。In some embodiments, the target gene is the ANGPTL3 gene. In some embodiments, the target portion of the sequence will be at least long enough to serve as a substrate for iRNA-directed cleavage at or near the nucleotide sequence portion of the mRNA molecule formed during transcription of the ANGPTL3 gene.
在本技术领域中，“G”、“C”、“A”、“T”和“U”通常分别代表鸟嘌呤、胞嘧啶、腺嘌呤、胸腺嘧啶、尿嘧啶的碱基，但本领域中也通常知晓，“G”、“C”、“A”、“T”和“U”每个通常也代表分别含有鸟嘌呤、胞嘧啶、腺嘌呤、胸腺嘧啶和尿嘧啶作为碱基的核苷酸，这在表示脱氧核糖核酸序列和/或核糖核酸序列中是常见的方式，因此在本公开的上下文中，“G”、“C”、“A”、“T”、“U”表示的含义包括上述各种可能的情形。然而，应理解术语“核糖核苷酸”或“核苷酸”还可以指一种经修饰的核苷酸(如以下进一步详述)或一种替代性的置换部分。本领域人员可以意识到，鸟嘌呤、胞嘧啶、腺嘌呤以及尿嘧啶可以被其他部分置换而基本上不改变一种寡核苷酸(包括一种具有这种置换部分的核苷酸)的碱基配对特性。例如非限制性地，包括肌苷作为其碱基的核苷酸可以与包括腺嘌呤、胞嘧啶或尿嘧啶的核苷酸进行碱基配对。因此，含有尿嘧啶、鸟嘌呤或腺嘌呤的核苷酸可以在本发明表征的dsRNA的核苷酸序列中由含有例如肌苷的核苷酸替换。在另一个实例中，寡核苷酸中任何地方的腺嘌呤和胞嘧啶可以分别地替换为鸟嘌呤和尿嘧啶，以形成与靶mRNA的G-U摇摆碱基配对。含有这类替换部分的序列适用于本发明表征的组合物和方法。In this technical field, "G", "C", "A", "T" and "U" usually represent the bases of guanine, cytosine, adenine, thymine and uracil respectively, but in this field It is also commonly known that "G", "C", "A", "T" and "U" each also generally represent nucleosides containing guanine, cytosine, adenine, thymine and uracil as bases respectively acid, which is a common way of expressing deoxyribonucleic acid sequences and/or ribonucleic acid sequences, so in the context of this disclosure, "G", "C", "A", "T", "U" The meaning includes various possible situations mentioned above. However, it is understood that the term "ribonucleotide" or "nucleotide" may also refer to a modified nucleotide (as described in further detail below) or an alternative replacement moiety. One skilled in the art will appreciate that guanine, cytosine, adenine, and uracil can be substituted by other moieties without substantially changing the base of an oligonucleotide, including a nucleotide having such a substituted moiety. Base pairing properties. For example, without limitation, a nucleotide including inosine as its base may be base-paired with a nucleotide including adenine, cytosine, or uracil. Thus, uracil, guanine or adenine containing nucleotides may be replaced by eg inosine containing nucleotides in the nucleotide sequence of the dsRNA characterized in the present invention. In another example, adenine and cytosine anywhere in the oligonucleotide can be replaced with guanine and uracil, respectively, to form G-U wobble base pairing with the target mRNA. Sequences containing such substituted moieties are suitable for use in the compositions and methods characterized herein.
本公开上下文使用的术语“iRNA”、“RNAi试剂”、“iRNA试剂”、“RNA干扰剂”在此可互换使用，是指在此所定义的术语包含siRNA，并且介导通过RNA诱导沉默复合物(RISC)途径的RNA转录物靶向切割。iRNA通过已知为RNA干扰(RNAi)的过程指导mRNA的序列特异性降解。iRNA调节，例如抑制，靶基因在细胞(如受试者(如哺乳动物受试者)的细胞)中的表达。The terms "iRNA," "RNAi agent," "iRNA agent," and "RNA interference agent" as used in the context of this disclosure are used interchangeably herein to mean that the terms as defined herein include siRNA and mediate silencing induced by RNA. Targeted cleavage of RNA transcripts by the RISC complex (RISC) pathway. iRNA directs the sequence-specific degradation of mRNA through a process known as RNA interference (RNAi). An iRNA modulates, eg, inhibits, the expression of a target gene in a cell, such as a cell of a subject (eg, a mammalian subject).
本公开上下文使用的术语“双链核糖核酸”、“双链RNA(dsRNA)分子”、“dsRNA”可以互换地使用。术语“dsRNA”，是指核糖核酸分子的复合体，其具有双链结构，包含两条反向平行的和基本上互补的核酸链，被称为相对于靶基因，例如ANGPTL3基因，具有“正义”和“反义”定向。在一些实施例中，双链核糖核酸(dsRNA)通过转录后基因沉默机制(在此称为RNA干扰或RNAi)触发靶RNA例如mRNA的降解。The terms "double-stranded ribonucleic acid," "double-stranded RNA (dsRNA) molecule," and "dsRNA" are used interchangeably in the context of this disclosure. The term "dsRNA" refers to a complex of ribonucleic acid molecules that has a double-stranded structure, containing two antiparallel and substantially complementary nucleic acid strands, and is said to have "sense" relative to a target gene, such as the ANGPTL3 gene. ” and “antonymy” orientation. In some embodiments, double-stranded ribonucleic acid (dsRNA) triggers the degradation of target RNA, such as mRNA, through a post-transcriptional gene silencing mechanism (referred to herein as RNA interference or RNAi).
通常，dsRNA分子的每条链的大部分的核苷酸是核糖核苷酸，但是如在此详述的，两条链的每一者或两者还可以包括一个或多个非核糖核苷酸，例如，脱氧核糖核苷酸和/或修饰的核苷酸。另外，如本公开中所用，“双链核糖核酸”可以包括具有化学修饰的核糖核苷酸、磷酸骨架等等。这些修饰可以包括在此披露的或在本领域中已知的所有类型的修饰。Typically, the majority of the nucleotides of each strand of a dsRNA molecule are ribonucleotides, but as detailed herein, each or both strands may also include one or more non-ribonucleotides Acids, for example, deoxyribonucleotides and/or modified nucleotides. Additionally, as used in this disclosure, "double-stranded ribonucleic acid" may include ribonucleotides with chemical modifications, phosphate backbones, and the like. These modifications may include all types of modifications disclosed herein or known in the art.
本公开上下文使用的术语“异核苷酸”是指核苷酸中碱基在核糖环上的位置发生改 变而形成的化合物，例如，碱基不与核糖环的1’-位相连，而是与核糖环的2’-位或3’-位相连而形成的化合物。The term "isonucleotide" as used in the context of this disclosure refers to a nucleotide in which the position of the bases on the ribose ring has been modified. For example, the base is not connected to the 1'-position of the ribose ring, but is connected to the 2'-position or 3'-position of the ribose ring.
在一些实施方案中，本公开的双链核糖核酸是siRNA，其与靶基因转录的mRNA序列(例如C基因转录的mRNA序列)相互作用以指导靶RNA的切割。不希望受理论约束，引入细胞中的长双链RNA被称作Dicer的III型核酸内切酶分解成siRNA(夏普(Sharp)等人，《基因与发育》(Genes Dev.)2001，15:485)。Dicer(核糖核酸酶III样酶)将dsRNA加工成至具有特征性双碱基3'突出端的19-23碱基对短干扰性RNA(Bernstein等人，(2001)自然(Nature)409:363)。这些siRNA随后掺入RNA诱导性沉默复合物(RISC)中，在其中一种或多种解旋酶解开siRNA双链体，这使得互补性反义链指导靶识别成为可能(Nykanen等人，(2001)细胞(Cell)107:309)。一旦与适宜的靶mRNA结合，RISC内部的一种或多种核酸内切酶切割靶以诱导沉默(巴希尔(Elbashir)等人，(2001)《基因与发育》(Genes Dev.)15:188)。In some embodiments, the double-stranded ribonucleic acid of the present disclosure is an siRNA that interacts with the mRNA sequence transcribed by the target gene (eg, the mRNA sequence transcribed by the C gene) to direct cleavage of the target RNA. Without wishing to be bound by theory, long double-stranded RNA introduced into cells is broken down into siRNA by a type III endonuclease called Dicer (Sharp et al., Genes Dev. 2001, 15: 485). Dicer (ribonuclease III-like enzyme) processes dsRNA into 19-23 base pair short interfering RNA with a characteristic two-base 3' overhang (Bernstein et al. (2001) Nature 409:363) . These siRNAs are subsequently incorporated into the RNA-induced silencing complex (RISC), where one or more helicases unwind the siRNA duplex, making it possible for the complementary antisense strand to guide target recognition (Nykanen et al., (2001) Cell 107:309). Once bound to the appropriate target mRNA, one or more endonucleases within the RISC cleave the target to induce silencing (Elbashir et al., (2001) Genes Dev. 15: 188).
本公开上下文使用的术语“突出的核苷酸”是指当双链核糖核酸的一条链的一个3’端延伸超出另一条链的5’端时从该dsRNA的双链体结构突出的一个或多个不成对的核苷酸，或反之亦然。“平端”或“平末端”意指在该双链核糖核酸的那端处不存在不成对的核苷酸，即无核苷酸突出端。一种“平末端的”双链核糖核酸是一种在其整个长度上都是双链、即在该分子的任一端处都无核苷酸突出端的dsRNA。The term "overhanging nucleotide" as used in the context of this disclosure refers to a protruding or Multiple unpaired nucleotides, or vice versa. "Blunt end" or "blunt end" means that there are no unpaired nucleotides, ie, no nucleotide overhangs, at that end of the double-stranded ribonucleic acid. A "blunt-ended" double-stranded RNA is a dsRNA that is double-stranded throughout its length, ie, it has no nucleotide overhangs at either end of the molecule.
术语“反义链”是指双链核糖核酸中与靶标序列(例如，来源于人类ANGPTL3mRNA)基本上互补的一个区域的链。在该互补性区域不与该靶标序列完全互补的情况下，错配在末端区域是最为可容忍的，并且如果出现错配，它们通常在末端的一个或多个区域，例如5’和/或3’末端的5、4、3、2或1个核苷酸之内。The term "antisense strand" refers to the strand of a region of double-stranded ribonucleic acid that is substantially complementary to a target sequence (eg, derived from human ANGPTL3 mRNA). In the case where the region of complementarity is not completely complementary to the target sequence, mismatches are most tolerated in the terminal region, and if mismatches occur, they are usually in one or more regions at the terminal end, such as the 5' and/or Within 5, 4, 3, 2 or 1 nucleotide of the 3' end.
术语“正义链”指的双链核糖核酸中含有与反义链区域基本上互补的区域的核酸链。The term "sense strand" refers to a double-stranded nucleic acid strand containing a region that is substantially complementary to a region of the antisense strand.
术语“互补”或“反向互补”一词可互相替代使用，并具有本领域技术人员周知的含义，即，在双链核酸分子中，一条链的碱基与另一条链上的碱基以互补的方式相配对。在DNA中，嘌呤碱基腺嘌呤(A)始终与嘧啶碱基胸腺嘧啶(T)(或者在RNA中为尿嘧啶(U))相配对；嘌呤碱基鸟嘌呤(C)始终与嘧啶碱基胞嘧啶(G)相配对。每个碱基对都包括一个嘌呤和一个嘧啶。当一条链上的腺嘌呤始终与另一条链上的胸腺嘧啶(或尿嘧啶)配对，以及鸟嘌呤始终与胞嘧啶配对时，两条链被认为是彼此相互补的，以及从其互补链的序列中可以推断出该链的序列。与此相应地，“错配”在本领域中意指在双链核酸中，对应位置上的碱基并未以互补的形式配对存在。The terms "complementary" or "reverse complement" are used interchangeably and have the meaning well known to those skilled in the art, that is, in a double-stranded nucleic acid molecule, the bases on one strand are connected to the bases on the other strand. Paired in complementary ways. In DNA, the purine base adenine (A) always pairs with the pyrimidine base thymine (T) (or uracil (U) in RNA); the purine base guanine (C) always pairs with the pyrimidine base Pairs with cytosine (G). Each base pair consists of a purine and a pyrimidine. When adenine on one strand always pairs with thymine (or uracil) on the other strand, and guanine always pairs with cytosine, the two strands are said to be complementary to each other, and from their complementary strands The sequence of the chain can be inferred from the sequence. Accordingly, "mismatch" in the art means that in double-stranded nucleic acids, the bases at corresponding positions do not pair in a complementary manner.
术语“基本上反向互补”是指所涉及的两段核苷酸序列之间存在不多于3个的碱基错配，即所涉及的两段核苷酸序列之间存在1个、2个或3个的碱基错配；“完全互补”是指两段核苷酸序列之间不存在碱基错配。The term "substantially reverse complementary" means that there are no more than 3 base mismatches between the two nucleotide sequences involved, that is, there are 1, 2, and 2 base mismatches between the two nucleotide sequences involved. or 3 base mismatches; "complete complementarity" means that there are no base mismatches between the two nucleotide sequences.
术语“互补”、“完全互补”和“基本上互补”可相对于在dsRNA的正义链与反义链之间，或dsRNA的反义链与靶标序列之间的碱基配对使用，如将从其使用的上下文理解。The terms "complementary", "completely complementary" and "substantially complementary" may be used with respect to base pairing between the sense and antisense strands of a dsRNA, or between the antisense strand of a dsRNA and a target sequence, as will be used from Contextual understanding of its use.
术语“抑制”，可以与“减少”、“沉默”、“下调”、“压制”和其他类似术语交替使用，并且包括任何水平的抑制。The term "suppression," may be used interchangeably with "reduction," "silencing," "downregulation," "suppression" and other similar terms and includes any level of suppression.
术语“抑制ANGPTL3基因的表达”包括抑制任何ANGPTL3基因(如例如小鼠ANGPTL3基因、大鼠ANGPTL3基因、猴ANGPTL3基因、或人类ANGPTL3基因)以及ANGPTL3基因的变体(例如天然存在的变体)或突变体的表达。因此，该ANGPTL3基因可以是野生型ANGPTL3基因、突变ANGPTL3基因、或在遗传操作的细胞、细胞群组或生物体的情形下的转基因ANGPTL3基因。The term "inhibiting expression of an ANGPTL3 gene" includes inhibiting any ANGPTL3 gene (eg, mouse ANGPTL3 gene, rat ANGPTL3 gene, monkey ANGPTL3 gene, or human ANGPTL3 gene) as well as variants (eg, naturally occurring variants) of the ANGPTL3 gene, or Expression of mutants. Thus, the ANGPTL3 gene may be a wild-type ANGPTL3 gene, a mutant ANGPTL3 gene, or in the case of a genetically manipulated cell, cell population, or organism, a transgenic ANGPTL3 gene.
“抑制ANGPTL3基因表达”包括任何水平的ANGPTL3基因的抑制，例如至少部分抑制ANGPTL3基因的表达，如抑制至少约5％、至少约10％、至少约15％、至少约 20％、至少约25％、至少约30％、至少约35％、至少约40％、至少约45％、至少约50％、至少约55％、至少约60％、至少约65％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％、至少约91％、至少约92％、至少约93％、至少约94％、至少约95％、至少约96％、至少约97％、至少约98％、或至少约99％。"Inhibition of ANGPTL3 gene expression" includes any level of inhibition of the ANGPTL3 gene, such as at least partial inhibition of the expression of the ANGPTL3 gene, such as inhibition of at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%.
术语“各自独立地”是指结构中存在的取值范围相同或相近的至少两个基团(或环系)可以在特定情形下具有相同或不同的含义。例如，取代基X和取代基Y各自独立地为氢、羟基、烷基或芳基，则当取代基X为氢时，取代基Y既可以为氢，也可以为羟基、烷基或芳基；同理，当取代基Y为氢时，取代基X既可以为氢，也可以为羟基、烷基或芳基。The term "each independently" means that at least two groups (or ring systems) present in the structure with the same or similar value ranges can have the same or different meanings under specific circumstances. For example, substituent X and substituent Y are each independently hydrogen, hydroxyl, alkyl or aryl, then when substituent X is hydrogen, substituent Y can be either hydrogen, hydroxyl, alkyl or aryl. ;Similarly, when the substituent Y is hydrogen, the substituent X can be either hydrogen, hydroxyl, alkyl or aryl.
术语“烷基”包括直链、支链或环状的饱和烷基。例如，烷基包括但不限于甲基、乙基、丙基、环丙基、正丁基、异丁基、仲丁基、叔丁基、环丁基、正戊基、环已基等类似基团。示例性的，“C1-6烷基”中的“C1-6”是指包含有1、2、3、4、5或6个碳原子的直链、支链或环状形式排列的基团。The term "alkyl" includes straight-chain, branched or cyclic saturated alkyl groups. For example, alkyl groups include, but are not limited to, methyl, ethyl, propyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl, n-pentyl, cyclohexyl, and the like. group. For example, "C1-6" in "C1-6 alkyl" refers to a group containing 1, 2, 3, 4, 5 or 6 carbon atoms arranged in a linear, branched or cyclic form. .
术语“烷氧基”在本文中是指烷基基团通过氧原子与分子其余部分相连(-O-烷基)，其中所述烷基如本文中所定义。烷氧基的非限制性实例包括甲氧基、乙氧基、三氟甲氧基、二氟甲氧基、正丙氧基、异丙氧基、正丁氧基、叔丁氧基、正戊氧基等。The term "alkoxy" as used herein refers to an alkyl group attached to the remainder of the molecule through an oxygen atom (-O-alkyl), wherein said alkyl group is as defined herein. Non-limiting examples of alkoxy include methoxy, ethoxy, trifluoromethoxy, difluoromethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n- Pentyloxy etc.
术语“治疗”是指：在罹患疾病之后，使受试者接触(例如给药)双链核糖核酸、双链核糖核酸修饰物、双链核糖核酸缀合物、药物组合物，从而与不接触时相比使该疾病的症状减轻，并不意味着必需完全抑制疾病的症状。罹患疾病是指：身体出现了疾病症状。The term "treatment" refers to exposing a subject to (e.g., administering) a double-stranded ribonucleic acid, a double-stranded ribonucleic acid modification, a double-stranded ribonucleic acid conjugate, or a pharmaceutical composition after suffering from a disease, so that the subject is not exposed to the disease. Reducing the symptoms of the disease does not necessarily mean that they must be completely suppressed. Suffering from a disease means that the body has symptoms of the disease.
术语“预防”是指：在罹患疾病之前，通过使受试者接触(例如给药)本公开的双链核糖核酸、双链核糖核酸修饰物、双链核糖核酸缀合物、药物组合物，从而与不接触时相比减轻罹患疾病后的症状，并不意味着必需完全抑制患病。The term "prevention" refers to: by exposing a subject to (e.g., administering) the double-stranded ribonucleic acid, double-stranded ribonucleic acid modification, double-stranded ribonucleic acid conjugate, pharmaceutical composition of the present disclosure before suffering from the disease, Thus, reducing the symptoms of a disease compared with no exposure does not necessarily mean that the disease must be completely suppressed.
术语“有效量”指本发明的双链核糖核酸、双链核糖核酸修饰物、双链核糖核酸缀合物或药物组合物的这样的量或剂量，其以单一或多次剂量施用患者后，在需要治疗或预防的患者中产生预期效果。有效量可以由作为本领域技术人员的主治医师通过考虑以下多种因素来容易地确定：诸如哺乳动物的物种；它的大小、年龄和一般健康；涉及的具体疾病；疾病的程度或严重性；个体患者的应答；施用的具体抗体；施用模式；施用制剂的生物利用率特征；选择的给药方案；和任何伴随疗法的使用。The term "effective amount" refers to an amount or dosage of the double-stranded ribonucleic acid, double-stranded ribonucleic acid modification, double-stranded ribonucleic acid conjugate or pharmaceutical composition of the present invention, which after administration to a patient in single or multiple doses, Produce the desired effect in patients in need of treatment or prophylaxis. The effective amount can be readily determined by the attending physician, who is one of skill in the art, by considering various factors such as: the species of the mammal; its size, age and general health; the specific disease involved; the extent or severity of the disease; The individual patient's response; the specific antibody administered; the mode of administration; the bioavailability characteristics of the administered formulation; the selected dosing regimen; and the use of any concomitant therapy.
术语“与ANGPTL3基因异常表达相关的疾病”是与代谢紊乱相关联的疾病或障碍。术语“与ANGPTL3基因异常表达相关的疾病”包括将从减少ANGPTL3(即“ANGPTL3-相关性疾病”)表达受益的疾病、障碍或病症。此类疾病典型地与血浆甘油三酯(triglyceride,TG)、高密度脂蛋白(high-density lipoprotein,HDL)、低密度脂蛋白(low-density lipoprotein,LDL)以及总胆固醇水平(total cholesterol,TC)变化密切相关。ANGPTL3基因异常表达相关的疾病的非限制性实例包括乳糜微粒血症综合征、2型糖尿病、家族性部分脂肪营养不良、高胆固醇血症、家族性高胆固醇血症、高甘油三酯血症、非酒精性脂肪肝、动脉粥样硬化、高甘油三酯血症、膝盖损伤及骨关节炎、血脂异常及相关代谢疾病、家族性低β脂蛋白血症、高脂血症、肥胖症、原发性肾病综合征、冠心病、恶性肿瘤等(参见，例如，Minicocci等，Mutations in the ANGPTL3gene and familial combined hypolipidemia:a clinical and biochemical characterization.J Clin Endocrinol Metab,2012,97(7):E1266-1275；Kaplan等，Regulation of the angiopoietin-like protein 3gene by LXR.J Lipid Res,2003,44(1):136-143；成海鹏等，脂蛋白脂酶降解与代谢异常性疾病，中国动脉硬化杂志,2017,25(05):513-518；饶佳等，儿童原发性肾病综合征中血管生成素样蛋白3的表达，中华肾脏病杂志,2006,22(05):286-290；Stitziel等，ANGPTL3deficiency and protection against coronary artery disease.J Am Coll Cardiol,2017,69(16):2054-2063；Wang  PF等，Clinical significance of angiopoietin-like protein 3expression in patients with glioblastoma.Neoplasma,2016,63(1):93-98)。The term "disease associated with abnormal expression of ANGPTL3 gene" is a disease or disorder associated with a metabolic disorder. The term "disease associated with abnormal expression of the ANGPTL3 gene" includes diseases, disorders or conditions that would benefit from reduced expression of ANGPTL3 (i.e., "ANGPTL3-associated disease"). Such diseases are typically associated with plasma triglyceride (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), and total cholesterol (TC) levels. ) changes are closely related. Non-limiting examples of diseases associated with abnormal expression of the ANGPTL3 gene include chylomicronemia syndrome, type 2 diabetes, familial partial lipodystrophy, hypercholesterolemia, familial hypercholesterolemia, hypertriglyceridemia, Non-alcoholic fatty liver disease, atherosclerosis, hypertriglyceridemia, knee injury and osteoarthritis, dyslipidemia and related metabolic diseases, familial hypobeta lipoproteinemia, hyperlipidemia, obesity, protozoa Nephrotic syndrome, coronary heart disease, malignant tumors, etc. (See, for example, Minicocci et al., Mutations in the ANGPTL3gene and familial combined hypolipidemia: a clinical and biochemical characterization. J Clin Endocrinol Metab, 2012, 97(7): E1266-1275 ; Kaplan et al., Regulation of the angiopoietin-like protein 3gene by LXR.J Lipid Res, 2003, 44(1):136-143; Cheng Haipeng et al., Lipoprotein lipase degradation and abnormal metabolic diseases, Chinese Journal of Arteriosclerosis, 2017,25(05):513-518; Rao Jia et al., Expression of angiopoietin-like protein 3 in children with primary nephrotic syndrome, Chinese Journal of Nephrology, 2006, 22(05):286-290; Stitziel et al. , ANGPTL3deficiency and protection against coronary artery disease.J Am Coll Cardiol, 2017,69(16):2054-2063; Wang PF et al., Clinical significance of angiopoietin-like protein 3expression in patients with glioblastoma.Neoplasma, 2016, 63(1):93-98).
术语“药学上可接受的辅料”或“药学上可接受的载体”是指在药物生产领域中广泛采用的辅助物料。使用辅料的主要目的在于提供一种使用安全、性质稳定和/或具有特定功能性的药物组合物，还在于提供一种方法，以便在为受试者施用药物之后，活性成分能够以所期望的速率溶出，或者促进活性成分在接受给药的受试者体内得到有效吸收。药学上可接受的辅料可以是具有惰性的填充剂，也可以是为药用组合物提供某种功能(例如稳定组合物的整体pH值或防止组合物中活性成分的降解)的功效成分。药学上可接受的辅料的非限制性实例包括但不限于粘合剂、助悬剂、乳化剂、稀释剂(或填充剂)、成粒剂、胶粘剂、崩解剂、润滑剂、抗粘着剂、助流剂、润湿剂、胶凝剂、吸收延迟剂、溶解抑制剂、增强剂、吸附剂、缓冲剂、螯合剂、防腐剂、着色剂、矫味剂、甜味剂等。The term "pharmaceutically acceptable excipients" or "pharmaceutically acceptable carriers" refers to auxiliary materials widely used in the field of pharmaceutical production. The main purpose of using excipients is to provide a pharmaceutical composition that is safe to use, stable in nature and/or has specific functionality, and also to provide a method so that after the drug is administered to the subject, the active ingredient can be used in the desired manner. rate dissolution, or promote effective absorption of the active ingredient in the subject to whom it is administered. Pharmaceutically acceptable excipients may be inert fillers or functional ingredients that provide a certain function for the pharmaceutical composition (such as stabilizing the overall pH value of the composition or preventing the degradation of the active ingredients in the composition). Non-limiting examples of pharmaceutically acceptable excipients include, but are not limited to, binders, suspending agents, emulsifiers, diluents (or fillers), granulating agents, adhesives, disintegrants, lubricants, and anti-adhesive agents. , glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents, sweeteners, etc.
本公开中的药物组合物可以使用本领域技术人员已知的任何方法来制备。例如，常规混合、溶解、造粒、乳化、磨细、包封、包埋和/或冻干工艺。Pharmaceutical compositions of the present disclosure may be prepared using any method known to those skilled in the art. For example, conventional mixing, dissolving, granulating, emulsifying, grinding, encapsulating, embedding and/or lyophilizing processes.
在本公开中，施用途经能够以任何适用的方式进行变化或调整，以满足药物的性质、患者和医务人员的便利以及其它相关因素的需求。In the present disclosure, the methods of administration can be varied or adapted in any applicable manner to meet the needs of the nature of the drug, convenience of the patient and medical staff, and other relevant factors.
本公开上下文中使用的术语“个体”、“患者”或“受试者”包括哺乳动物。哺乳动物包括但不限于，家养动物(例如，牛，羊，猫，狗和马)，灵长类动物(例如，人和非人灵长类动物如猴)，兔，以及啮齿类动物(例如，小鼠和大鼠)。The terms "individual," "patient," or "subject" as used in the context of this disclosure include mammals. Mammals include, but are not limited to, domestic animals (e.g., cattle, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., , mice and rats).
除非另外定义或由背景清楚指示，否则在本公开中的全部技术与科学术语具有如本公开所属领域的普通技术人员通常理解的相同含义。Unless defined otherwise or clearly indicated by context, all technical and scientific terms in this disclosure have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.
双链核糖核酸double stranded ribonucleic acid
本公开的第一方面提供一种双链核糖核酸(dsRNA)，用于抑制ANGPTL3基因的表达。双链核糖核酸的一条链为反义链，反义链与靶基因(也即，ANGPTL3基因)在表达过程中形成的mRNA序列互补配对，用于指导靶mRNA(也即，ANGPTL3基因的转录产物)的切割。双链核糖核酸中另一条正义链包括与反义链部分互补和完全互补形成双链核糖核酸的双链区。A first aspect of the present disclosure provides a double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of ANGPTL3 gene. One strand of the double-stranded ribonucleic acid is the antisense strand. The antisense strand is complementary to the mRNA sequence formed during the expression of the target gene (i.e., ANGPTL3 gene) and is used to guide the transcription product of the target mRNA (i.e., ANGPTL3 gene). ) cutting. The other sense strand in the double-stranded RNA includes a double-stranded region that is partially complementary and fully complementary to the antisense strand to form a double-stranded RNA.
在一些实施方案中，双链核糖核酸作为核酸内切酶(Dicer)的底物，被切割为小片段的dsRNA，也即siRNA。在一些实施方案中，双链核糖核酸为siRNA。siRNA通过装配形成RNA诱导的沉默复合物(RNA-induced silencing complex，RISC)RISC复合体，切割靶mRNA，抑制ANGPTL3基因的表达。In some embodiments, double-stranded ribonucleic acid serves as a substrate for endonuclease (Dicer) and is cleaved into small fragments of dsRNA, that is, siRNA. In some embodiments, the double-stranded ribonucleic acid is siRNA. siRNA assembles to form the RNA-induced silencing complex (RISC) RISC complex, which cleaves the target mRNA and inhibits the expression of the ANGPTL3 gene.
根据来源于人ANGPTL3mRNA(NM_014495.4)中的靶标序列，设计与靶mRNA结合的siRNA。在一些实施方案中，靶标序列选自如SEQ ID NO:1-7任一项所示的核苷酸序列。在一些更为具体的实施方案中，靶标序列选自如SEQ ID NO:8-25任一项所示的核苷酸序列。Based on the target sequence derived from human ANGPTL3 mRNA (NM_014495.4), siRNA that binds to the target mRNA is designed. In some embodiments, the target sequence is selected from the nucleotide sequences set forth in any one of SEQ ID NOs: 1-7. In some more specific embodiments, the target sequence is selected from the nucleotide sequence set forth in any one of SEQ ID NOs: 8-25.
在一些具体的实施方案中，SEQ ID NO:1所示的核苷酸序列包含SEQ ID NO:8～10所示的核苷酸序列。In some specific embodiments, the nucleotide sequence shown in SEQ ID NO: 1 includes the nucleotide sequence shown in SEQ ID NO: 8-10.
在一些具体的实施方案中，SEQ ID NO:2所示的核苷酸序列包含SEQ ID NO:11～12所示的核苷酸序列。In some specific embodiments, the nucleotide sequence shown in SEQ ID NO:2 includes the nucleotide sequence shown in SEQ ID NO:11-12.
在一些具体的实施方案中，SEQ ID NO:3所示的核苷酸序列包含SEQ ID NO:13～14所示的核苷酸序列。In some specific embodiments, the nucleotide sequence shown in SEQ ID NO:3 includes the nucleotide sequence shown in SEQ ID NO:13-14.
在一些具体的实施方案中，SEQ ID NO:4所示的核苷酸序列包含SEQ ID NO:15～16所示的核苷酸序列。In some specific embodiments, the nucleotide sequence shown in SEQ ID NO:4 includes the nucleotide sequence shown in SEQ ID NO:15-16.
在一些具体的实施方案中，SEQ ID NO:5所示的核苷酸序列包含SEQ ID NO:17～19所示的核苷酸序列。In some specific embodiments, the nucleotide sequence shown in SEQ ID NO:5 includes the nucleotide sequence shown in SEQ ID NO:17～19.
在一些具体的实施方案中，SEQ ID NO:6所示的核苷酸序列包含SEQ ID NO:20～22 所示的核苷酸序列。In some specific embodiments, the nucleotide sequence shown in SEQ ID NO: 6 includes SEQ ID NO: 20-22 The nucleotide sequence shown.
在一些具体的实施方案中，SEQ ID NO:7所示的核苷酸序列包含SEQ ID NO:23～25所示的核苷酸序列。In some specific embodiments, the nucleotide sequence shown in SEQ ID NO:7 includes the nucleotide sequence shown in SEQ ID NO:23～25.
在一些实施方案中，反义链包含与靶标序列中至少15个连续核苷酸的反向互补序列的差异不超过3个核苷酸的序列B。具体地，沿5’末端向3’末端的方向，在靶标序列中选择起始核苷酸，以包含起始核苷酸在内的向3’方向延伸的至少15个核苷酸作为siRNA的结合区域。反义链包含结合区域对应的核苷酸序列的反向互补序列。需要说明的是，起始核苷酸可以是靶标序列任意位置处的核苷酸，只要基于该起始核苷酸向靶标序列3’方向延伸，可以得到至少15个连续核苷酸(包含起始位置处的核苷酸)即可。In some embodiments, the antisense strand comprises sequence B that differs by no more than 3 nucleotides from the reverse complement of at least 15 contiguous nucleotides in the target sequence. Specifically, the starting nucleotide is selected in the target sequence in the direction from the 5' end to the 3' end, and at least 15 nucleotides extending in the 3' direction including the starting nucleotide are used as the siRNA. Combined area. The antisense strand contains the reverse complement of the nucleotide sequence corresponding to the binding region. It should be noted that the starting nucleotide can be a nucleotide at any position of the target sequence, as long as at least 15 consecutive nucleotides (including the starting nucleotide) can be obtained based on the starting nucleotide extending in the 3' direction of the target sequence. nucleotide at the starting position).
在本公开中，反义链的核苷酸序列与靶标序列可以是完全互补或基本上互补。当反义链的核苷酸序列与靶标序列基本上互补时，反义链的核苷酸序列中存在与靶标序列存在不超过3个的错配碱基。例如，错配碱基为1个、2个或3个。当反义链的核苷酸序列与靶标序列完全互补时，反义链的核苷酸序列与靶标序列不存在错配碱基。In the present disclosure, the nucleotide sequence of the antisense strand may be completely complementary or substantially complementary to the target sequence. When the nucleotide sequence of the antisense strand is substantially complementary to the target sequence, the nucleotide sequence of the antisense strand contains no more than 3 mismatched bases with the target sequence. For example, there are 1, 2, or 3 mismatched bases. When the nucleotide sequence of the antisense strand is completely complementary to the target sequence, there are no mismatched bases between the nucleotide sequence of the antisense strand and the target sequence.
进一步地，反义链由至少15个核苷酸组成。在一些实施方案中，反义链由15-28个核苷酸组成。例如，反义链的长度为15、16、17、18、19、20、21、22、23、24、25、26、27或28个核苷酸。Further, the antisense strand consists of at least 15 nucleotides. In some embodiments, the antisense strand consists of 15-28 nucleotides. For example, the antisense strand is 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 nucleotides in length.
作为优选，反义链由19-25个核苷酸组成，更优选19-23个核苷酸，最优选19、21或23个核苷酸。Preferably, the antisense strand consists of 19-25 nucleotides, more preferably 19-23 nucleotides, and most preferably 19, 21 or 23 nucleotides.
在一些可选的实施方案中，反义链包含的序列B与靶标序列上至少15个连续核苷酸组成的序列的反向互补序列完全相同。In some alternative embodiments, the antisense strand contains sequence B that is identical to the reverse complement of a sequence consisting of at least 15 consecutive nucleotides on the target sequence.
在一些具体的实施方案中，反义链包含的序列B与靶标序列上15-28个连续核苷酸组成的序列的反向互补序列完全相同。优选靶标序列上19-25个连续核苷酸，更优选靶标序列上19-23个连续核苷酸，最优选19、21或23个连续核苷酸。In some specific embodiments, the antisense strand contains sequence B that is identical to the reverse complement of a sequence consisting of 15-28 consecutive nucleotides on the target sequence. Preferably, there are 19-25 contiguous nucleotides on the target sequence, more preferably 19-23 contiguous nucleotides on the target sequence, and most preferably 19, 21 or 23 contiguous nucleotides.
在一些可选的实施方案中，反义链包含的序列B与靶标序列上至少15个连续核苷酸组成的序列的反向互补序列存在1个核苷酸的差异。In some alternative embodiments, the antisense strand contains sequence B that differs by 1 nucleotide from the reverse complement of a sequence consisting of at least 15 consecutive nucleotides on the target sequence.
在一些具体的实施方案中，反义链包含的序列B与靶标序列上15-28个核苷酸组成的序列的反向互补序列存在1个核苷酸的差异。优选靶标序列上19-25个连续核苷酸，更优选靶标序列上19-23个连续核苷酸，最优选19、21或23个连续核苷酸。In some specific embodiments, the antisense strand contains sequence B that differs by 1 nucleotide from the reverse complement of a sequence consisting of 15-28 nucleotides on the target sequence. Preferably, there are 19-25 contiguous nucleotides on the target sequence, more preferably 19-23 contiguous nucleotides on the target sequence, and most preferably 19, 21 or 23 contiguous nucleotides.
在一些具体的实施方案中，差异的核苷酸位于序列B的3’末端。In some specific embodiments, the differential nucleotides are located at the 3' end of sequence B.
在一些实施方案中，所述正义链包含与靶标序列中至少15个连续核苷酸的差异不超过3个核苷酸的序列A。正义链中包括与反义链互补的区域，正义链的核苷酸序列与反义链在靶标序列上结合区域的序列完全相同或基本完全相同。因此，正义链的核苷酸序列为靶标序列中结合反义链的至少15个连续核苷酸；或者，正义链的核苷酸序列与靶标序列中结合反义链的至少15个连续核苷酸相比，存在1个、2个或3个碱基不同的差异核苷酸。In some embodiments, the sense strand comprises sequence A that differs by no more than 3 nucleotides from at least 15 consecutive nucleotides in the target sequence. The sense strand includes a region complementary to the antisense strand, and the nucleotide sequence of the sense strand is identical or substantially identical to the sequence of the binding region of the antisense strand on the target sequence. Therefore, the nucleotide sequence of the sense strand is at least 15 consecutive nucleotides in the target sequence that binds the antisense strand; alternatively, the nucleotide sequence of the sense strand is the same as at least 15 consecutive nucleotides in the target sequence that binds the antisense strand. Compared to acids, there are differential nucleotides that differ by 1, 2 or 3 bases.
进一步地，正义链由至少15个核苷酸组成。在一些实施方案中，正义链由15-28个核苷酸组成。例如，正义链的长度为15、16、17、18、19、20、21、22、23、24、25、26、27或28个核苷酸。Further, the sense strand consists of at least 15 nucleotides. In some embodiments, the sense strand consists of 15-28 nucleotides. For example, the sense strand is 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 nucleotides in length.
作为优选，正义链由19-25个核苷酸组成，更优选19-23个核苷酸，最优选19、21或23个核苷酸。Preferably, the sense strand consists of 19-25 nucleotides, more preferably 19-23 nucleotides, and most preferably 19, 21 or 23 nucleotides.
在一些可选的实施方案中，正义链包含的序列A与靶标序列上至少15个连续核苷酸组成的序列完全相同。In some alternative embodiments, the sense strand includes sequence A that is identical to the sequence consisting of at least 15 consecutive nucleotides on the target sequence.
在一些具体的实施方案中，正义链包含的序列A与靶标序列上15-28个连续核苷酸组成的序列完全相同。优选靶标序列上19-25个连续核苷酸，更优选靶标序列上19-23个连续核苷酸，最优选靶标序列上19、21或23个连续核苷酸。 In some specific embodiments, the sense strand contains sequence A that is identical to the sequence consisting of 15-28 consecutive nucleotides on the target sequence. Preferably, there are 19-25 contiguous nucleotides on the target sequence, more preferably 19-23 contiguous nucleotides on the target sequence, and most preferably 19, 21 or 23 contiguous nucleotides on the target sequence.
在一些可选的实施方案中，正义链包含的序列A与靶标序列上至少15个连续核苷酸组成的序列存在1个核苷酸的差异。In some alternative embodiments, the sense strand contains sequence A that differs by 1 nucleotide from a sequence consisting of at least 15 consecutive nucleotides on the target sequence.
在一些具体的实施方案中，正义链包含的序列A与靶标序列上15-28个连续核苷酸组成的序列存在1个核苷酸的差异。优选靶标序列上19-25个连续核苷酸，更优选靶标序列上19-23个连续核苷酸，最优选靶标序列上19、21或23个连续核苷酸。In some specific embodiments, the sense strand contains sequence A that differs by 1 nucleotide from the sequence consisting of 15-28 consecutive nucleotides on the target sequence. Preferably, there are 19-25 contiguous nucleotides on the target sequence, more preferably 19-23 contiguous nucleotides on the target sequence, and most preferably 19, 21 or 23 contiguous nucleotides on the target sequence.
在一些具体的实施方案中，差异的核苷酸位于序列A的3’末端。In some specific embodiments, the differential nucleotides are located at the 3' end of sequence A.
在本公开中，正义链的长度与反义链长度可以相同或不同。In this disclosure, the length of the sense strand and the length of the antisense strand may be the same or different.
在一些实施方式中，正义链与反义链的长度相同，具体地，正义链/反义链的长度比为15/15、16/16、17/17、18/18、19/19、20/20、21/21、22/22、23/23、24/24、25/25、26/26、27/27或28/28。作为优选，正义链/反义链的长度比为19/19、20/20、21/21、22/22、23/23、24/24或25/25，更优选19/19、20/20、21/21、22/22或23/23，最优选19/19、21/21或23/23；在一些优选的实施方式中，正义链/反义链的长度比为19/21或21/23。In some embodiments, the sense strand and the antisense strand have the same length. Specifically, the length ratio of the sense strand/antisense strand is 15/15, 16/16, 17/17, 18/18, 19/19, 20 /20, 21/21, 22/22, 23/23, 24/24, 25/25, 26/26, 27/27 or 28/28. Preferably, the length ratio of sense strand/antisense strand is 19/19, 20/20, 21/21, 22/22, 23/23, 24/24 or 25/25, more preferably 19/19, 20/20 , 21/21, 22/22 or 23/23, most preferably 19/19, 21/21 or 23/23; in some preferred embodiments, the length ratio of the sense strand/antisense strand is 19/21 or 21 /twenty three.
在一些实施方式中，正义链与反义链的长度不同。例如，正义链/反义链的长度比为19/20、19/21、19/22、19/23、19/24、19/25、19/26、20/19、20/21、20/22、20/23、20/24、20/25、20/26、21/19、21/20、21/22、21/23、21/24、21/25、21/26、22/19、22/20、22/21、22/23、22/24、22/25、22/26、23/19、23/20、23/21、23/22、23/24、23/25或23/26等等。In some embodiments, the sense and antisense strands are different lengths. For example, the length ratio of sense strand/antisense strand is 19/20, 19/21, 19/22, 19/23, 19/24, 19/25, 19/26, 20/19, 20/21, 20/ 22, 20/23, 20/24, 20/25, 20/26, 21/19, 21/20, 21/22, 21/23, 21/24, 21/25, 21/26, 22/19, 22/20, 22/21, 22/23, 22/24, 22/25, 22/26, 23/19, 23/20, 23/21, 23/22, 23/24, 23/25 or 23/ 26 and so on.
在本公开中，正义链与反义链可以是完全互补或基本上互补，当两者基本上互补时，正义链与反义链形成的双链区内存在不超过3个错配碱基。In the present disclosure, the sense strand and the antisense strand may be completely complementary or substantially complementary. When the two are substantially complementary, there will be no more than 3 mismatched bases in the double-stranded region formed by the sense strand and the antisense strand.
在一些实施方案中，正义链与反义链互补形成双链区后，正义链、反义链或其组合具有延伸出所述双链区的突出的核苷酸。突出的核苷酸的数量可以是1个或多个，例如，1个或2个。另外，突出1-2个核苷酸可以位于任意反义链或正义链的5’末端、3’末端或两端，并且，每一个突出的核苷酸可以是任意类型的核苷酸。In some embodiments, after the sense strand and the antisense strand are complementary to form a double-stranded region, the sense strand, the antisense strand, or a combination thereof has overhanging nucleotides extending out of the double-stranded region. The number of overhanging nucleotides may be 1 or more, for example, 1 or 2. In addition, the protruding 1-2 nucleotides can be located at the 5' end, 3' end or both ends of any antisense strand or sense strand, and each protruding nucleotide can be any type of nucleotide.
在一些实施方案中，所述正义链与所述反义链互补形成所述双链区，且所述正义链的3’末端具有1-2个延伸出所述双链区的突出的核苷酸，所述反义链的3’末端形成平末端。In some embodiments, the sense strand is complementary to the antisense strand to form the double-stranded region, and the 3' end of the sense strand has 1-2 protruding nucleosides extending out of the double-stranded region. acid, the 3' end of the antisense strand forms a blunt end.
在一些实施方案中，所述正义链与所述反义链互补形成所述双链区，且所述反义链的3’末端具有1-2个延伸出所述双链区的突出的核苷酸，所述正义链的3’末端形成平末端。In some embodiments, the sense strand and the antisense strand are complementary to form the double-stranded region, and the 3' end of the antisense strand has 1-2 protruding cores extending out of the double-stranded region. The 3' end of the sense strand forms a blunt end.
在一些实施方案中，所述正义链与所述反义链互补形成所述双链区，且所述正义链与所述反义链的3’末端均具有1-2个延伸出所述双链区的突出的核苷酸。In some embodiments, the sense strand and the antisense strand are complementary to form the double-stranded region, and the 3' ends of the sense strand and the antisense strand each have 1-2 strands extending out of the double-stranded region. Overhanging nucleotides in the chain region.
在一些实施方案中，所述正义链与所述反义链互补形成所述双链区，且所述正义链与所述反义链的3’末端均形成平末端。In some embodiments, the sense strand and the antisense strand are complementary to form the double-stranded region, and the 3' ends of the sense strand and the antisense strand both form blunt ends.
在本公开中，如上所述，反义链的核苷酸序列与靶标序列可以是完全互补或基本上互补；正义链与反义链可以是完全互补或基本上互补。因此，以下对于靶标序列SEQ ID NO:8～SEQ ID NO:25以及能够与这些靶标序列互补的siRNA的描述，对于每个siRNA的反义链，均包括与其互补的靶标序列(例如，SEQ ID NO:8～SEQ ID NO:25中的任一者)基本上互补的情况，也即，每个siRNA的反义链的核苷酸序列中可以存在与相应的靶标序列存在碱基错配的情况；对于每个siRNA的正义链，均包括与其互补的靶标序列(例如，SEQ ID NO:8～SEQ ID NO:25中的任一者)基本上互补的情况，也即，每个siRNA的正义链的核苷酸序列中可以存在与相应的靶标序列存在碱基错配的情况。在一些实施方案中，所述碱基错配可以是与靶标序列的差异不超过3个碱基的错配，例如，错配碱基为1个、2个或3个。需要说明的是，对于靶标序列SEQ ID NO:1～SEQ ID NO:7中的任一者，其也适用上述对于靶标序列SEQ ID NO:8～SEQ ID NO:25和与之互补的siRNA的碱基错配的情况的描述；也即，每个siRNA的反义链的核苷酸序列中可以存在与相应的靶标序列存在碱基错配的情况，每个siRNA的正义链的核苷酸序列中可以存在与相应的靶标序列存在碱基错配的情况。在一些实施方案中，所述碱基错配可以是与靶标序列的差异不超过3个碱基的错配，例如，错配碱基为1个、2个或3个。 In the present disclosure, as mentioned above, the nucleotide sequence of the antisense strand and the target sequence may be completely complementary or substantially complementary; the sense strand and the antisense strand may be completely complementary or substantially complementary. Therefore, the following description of the target sequences SEQ ID NO: 8 to SEQ ID NO: 25 and siRNA that can be complementary to these target sequences, for the antisense strand of each siRNA, includes the target sequence that is complementary to it (for example, SEQ ID Any one of NO:8 to SEQ ID NO:25) is substantially complementary, that is, the nucleotide sequence of the antisense strand of each siRNA may have a base mismatch with the corresponding target sequence. Situation; For the sense strand of each siRNA, it includes a situation that is substantially complementary to its complementary target sequence (for example, any one of SEQ ID NO: 8 ~ SEQ ID NO: 25), that is, the sense strand of each siRNA The nucleotide sequence of the sense strand may have base mismatches with the corresponding target sequence. In some embodiments, the base mismatch may be a mismatch that differs from the target sequence by no more than 3 bases, for example, the mismatched base is 1, 2, or 3 bases. It should be noted that for any one of the target sequences SEQ ID NO: 1 to SEQ ID NO: 7, the above-mentioned method for the target sequences SEQ ID NO: 8 to SEQ ID NO: 25 and siRNA complementary thereto is also applicable. Description of base mismatch situations; that is, the nucleotide sequence of the antisense strand of each siRNA may have base mismatches with the corresponding target sequence, and the nucleotide sequence of the sense strand of each siRNA There may be base mismatches in the sequence with the corresponding target sequence. In some embodiments, the base mismatch may be a mismatch that differs from the target sequence by no more than 3 bases, for example, the mismatched base is 1, 2, or 3 bases.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:8所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:8所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:8, and the nucleotide sequence of the antisense strand is SEQ ID NO:8 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:8所示序列中23个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:8所示序列中23个连续核苷酸组成的序列B组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 23 consecutive nucleotides in the sequence shown in SEQ ID NO:8, and the nucleotide sequence of the antisense strand is SEQ ID NO:8 The sequence B consisting of 23 consecutive nucleotides in the sequence shown is the reverse complementary sequence B. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
示例性的，双链核糖核酸为表1或表1-1中示出的siRNA1～siRNA3、siRNA87～siRNA88中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA1 to siRNA3 and siRNA87 to siRNA88 shown in Table 1 or Table 1-1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:9所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:9所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:9, and the nucleotide sequence of the antisense strand is SEQ ID NO:9 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
示例性的，双链核糖核酸为表1或表1-1中示出的siRNA4、siRNA89中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any one of siRNA4 and siRNA89 shown in Table 1 or Table 1-1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:10所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:10所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:10, and the nucleotide sequence of the antisense strand is SEQ ID NO:10 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
示例性的，双链核糖核酸为表1中示出的siRNA5。Exemplarily, the double-stranded ribonucleic acid is siRNA5 shown in Table 1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:11所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:11所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:11, and the nucleotide sequence of the antisense strand is SEQ ID NO:11 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:11所示序列中23个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:11所示序列中23个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 23 consecutive nucleotides in the sequence shown in SEQ ID NO:11, and the nucleotide sequence of the antisense strand is SEQ ID NO:11 The reverse complementary sequence B consisting of 23 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
示例性的，双链核糖核酸为表1中示出的siRNA6～siRNA7中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any one of siRNA6 to siRNA7 shown in Table 1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:12所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:12所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:12, and the nucleotide sequence of the antisense strand is SEQ ID NO:12 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
示例性的，双链核糖核酸为表1或表1-1中示出的siRNA8、siRNA120～siRNA121中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any one of siRNA8, siRNA120 to siRNA121 shown in Table 1 or Table 1-1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:13所示序列中23个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:13所示序列中23个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 23 consecutive nucleotides in the sequence shown in SEQ ID NO:13, and the nucleotide sequence of the antisense strand is SEQ ID NO:13 The reverse complementary sequence B consisting of 23 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:13所示序列中21个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:13所示序列中21个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 21 consecutive nucleotides in the sequence shown in SEQ ID NO:13, and the nucleotide sequence of the antisense strand is SEQ ID NO:13 Sequence B which is the reverse complement of the sequence consisting of 21 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:13所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:13所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:13, and the nucleotide sequence of the antisense strand is SEQ ID NO:13 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
示例性的，双链核糖核酸为表1或表1-1中示出的siRNA9～siRNA15、siRNA90～siRNA91中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA9 to siRNA15 and siRNA90 to siRNA91 shown in Table 1 or Table 1-1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:14所示序列中23个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:14所示序列中23个连续核苷 酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 23 consecutive nucleotides in the sequence shown in SEQ ID NO:14, and the nucleotide sequence of the antisense strand is SEQ ID NO:14 23 consecutive nucleotides in the sequence shown The acid sequence consists of the reverse complement of sequence B. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:14所示序列中21个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:14所示序列中21个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 21 consecutive nucleotides in the sequence shown in SEQ ID NO:14, and the nucleotide sequence of the antisense strand is SEQ ID NO:14 The reverse complementary sequence B consisting of 21 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:14所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:14所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:14, and the nucleotide sequence of the antisense strand is SEQ ID NO:14 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
示例性的，双链核糖核酸为表1或表1-1中示出的siRNA16～siRNA25、siRNA122～siRNA 124中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA16 to siRNA25 and siRNA122 to siRNA 124 shown in Table 1 or Table 1-1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:15所示序列中21个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:15所示序列中21个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 21 consecutive nucleotides in the sequence shown in SEQ ID NO:15, and the nucleotide sequence of the antisense strand is SEQ ID NO:15 The reverse complementary sequence B consisting of 21 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:15所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:15所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:15, and the nucleotide sequence of the antisense strand is SEQ ID NO:15 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
示例性的，双链核糖核酸为表1中示出的siRNA26～siRNA30中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA26 to siRNA30 shown in Table 1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:16所示序列中23个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:16所示序列中23个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 23 consecutive nucleotides in the sequence shown in SEQ ID NO:16, and the nucleotide sequence of the antisense strand is SEQ ID NO:16 The reverse complementary sequence B consisting of 23 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:16所示序列中21个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:16所示序列中21个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 21 consecutive nucleotides in the sequence shown in SEQ ID NO:16, and the nucleotide sequence of the antisense strand is SEQ ID NO:16 The reverse complementary sequence B consisting of 21 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:16所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:16所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:16, and the nucleotide sequence of the antisense strand is SEQ ID NO:16 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
示例性的，双链核糖核酸为表1中示出的siRNA31～siRNA40中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA31 to siRNA40 shown in Table 1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:17所示序列中23个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:17所示序列中23个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 23 consecutive nucleotides in the sequence shown in SEQ ID NO:17, and the nucleotide sequence of the antisense strand is SEQ ID NO:17 The reverse complementary sequence B consisting of 23 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:17所示序列中21个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:17所示序列中21个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 21 consecutive nucleotides in the sequence shown in SEQ ID NO:17, and the nucleotide sequence of the antisense strand is SEQ ID NO:17 The reverse complementary sequence B consisting of 21 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:17所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:17所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:17, and the nucleotide sequence of the antisense strand is SEQ ID NO:17 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
示例性的，双链核糖核酸为表1或表1-1中示出的siRNA41～siRNA55、siRNA92、siRNA125中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA41 to siRNA55, siRNA92, and siRNA125 shown in Table 1 or Table 1-1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:18所示序列中19个连 续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:18所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is the 19 consecutive nucleotides in the sequence shown in SEQ ID NO:18. The nucleotide sequence of the antisense strand is Sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO: 18, which is the reverse complement of Sequence B. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
示例性的，双链核糖核酸为表1中示出的siRNA56。Exemplarily, the double-stranded ribonucleic acid is siRNA56 shown in Table 1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:19所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:19所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:19, and the nucleotide sequence of the antisense strand is SEQ ID NO:19 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
在一些具体的实施方式中，正义链的核苷酸序列为与SEQ ID NO:19所示序列中19个连续核苷酸组成的序列相比存在一个差异的核苷酸的序列A，反义链的核苷酸序列为SEQ ID NO:19所示序列中19个连续核苷酸组成的序列的反向互补序列相比存在一个差异的核苷酸的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is a sequence A of nucleotides with one difference compared to the sequence consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:19, antisense The nucleotide sequence of the chain is the reverse complementary sequence of the sequence consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:19. Compared with the sequence B, there is one different nucleotide. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
示例性的，双链核糖核酸为表1中示出的siRNA57～siRNA58中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA57 to siRNA58 shown in Table 1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:20所示序列中23个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:20所示序列中23个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 23 consecutive nucleotides in the sequence shown in SEQ ID NO:20, and the nucleotide sequence of the antisense strand is SEQ ID NO:20 The reverse complementary sequence B consisting of 23 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:20所示序列中21个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:20所示序列中21个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 21 consecutive nucleotides in the sequence shown in SEQ ID NO:20, and the nucleotide sequence of the antisense strand is SEQ ID NO:20 Sequence B which is the reverse complement of the sequence consisting of 21 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:20所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:20所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:20, and the nucleotide sequence of the antisense strand is SEQ ID NO:20 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
示例性的，双链核糖核酸为表1中示出的siRNA59～siRNA63中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA59 to siRNA63 shown in Table 1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:21所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:21所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:21, and the nucleotide sequence of the antisense strand is SEQ ID NO:21 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
示例性的，双链核糖核酸为表1或表1-1中示出的siRNA64、siRNA126中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any one of siRNA64 and siRNA126 shown in Table 1 or Table 1-1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:22所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:22所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:22, and the nucleotide sequence of the antisense strand is SEQ ID NO:22 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
在一些具体的实施方式中，正义链的核苷酸序列为与SEQ ID NO:22所示序列中19个连续核苷酸组成的序列相比存在一个差异的核苷酸的序列A，反义链的核苷酸序列为SEQ ID NO:22所示序列中19个连续核苷酸组成的序列的反向互补序列相比存在一个差异的核苷酸的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is a sequence A of nucleotides with one difference compared to the sequence consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:22, antisense The nucleotide sequence of the chain is the reverse complementary sequence of the sequence consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:22. Compared with the sequence B, there is one different nucleotide. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:22所示序列中21个连续核苷酸组成的序列A或与序列A存在一个核苷酸差异的序列，反义链的核苷酸序列为SEQ ID NO:22所示序列中21个连续核苷酸组成的序列反向互补的序列B或与序列B存在一个核苷酸差异的序列。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 21 consecutive nucleotides in the sequence shown in SEQ ID NO: 22 or a sequence that is one nucleotide different from sequence A, antisense The nucleotide sequence of the chain is the reverse complementary sequence B consisting of 21 consecutive nucleotides in the sequence shown in SEQ ID NO:22 or a sequence that differs by one nucleotide from sequence B. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:22所示序列中23个连续核苷酸组成的序列A或与序列A存在一个核苷酸差异的序列，反义链的核苷酸序列为SEQ ID NO:22所示序列中23个连续核苷酸组成的序列反向互补的序列B或与序列B存在一个核苷酸差异的序列。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 23 consecutive nucleotides in the sequence shown in SEQ ID NO: 22 or a sequence that is one nucleotide different from sequence A, antisense The nucleotide sequence of the chain is the reverse complementary sequence B consisting of 23 consecutive nucleotides in the sequence shown in SEQ ID NO:22 or a sequence that differs by one nucleotide from sequence B. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
示例性的，双链核糖核酸为表1或表1-1中示出的siRNA65～siRNA73、siRNA93、 siRNA127～siRNA128中的任一siRNA。Exemplarily, the double-stranded ribonucleic acids are siRNA65 to siRNA73, siRNA93, and siRNA shown in Table 1 or Table 1-1. Any siRNA among siRNA127 to siRNA128.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:23所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:23所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:23, and the nucleotide sequence of the antisense strand is SEQ ID NO:23 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
示例性的，双链核糖核酸为表1中示出的siRNA74～siRNA76中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA74 to siRNA76 shown in Table 1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:24所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:24所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:24, and the nucleotide sequence of the antisense strand is SEQ ID NO:24 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
在一些具体的实施方式中，正义链的核苷酸序列为与SEQ ID NO:24所示序列中19个连续核苷酸组成的序列相比存在一个差异的核苷酸的序列A，反义链的核苷酸序列为与SEQ ID NO:24所示序列中19个连续核苷酸组成的序列的反向互补序列相比存在一个差异的核苷酸的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is a sequence A of nucleotides with one difference compared to the sequence consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:24, antisense The nucleotide sequence of the chain is sequence B with one nucleotide difference compared to the reverse complement sequence of the sequence consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:24. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:24所示序列中21个连续核苷酸组成的序列A或与序列A存在一个核苷酸差异的序列，反义链的核苷酸序列为SEQ ID NO:24所示序列中21个连续核苷酸组成的序列反向互补的序列B或与序列B存在一个核苷酸差异的序列。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 21 consecutive nucleotides in the sequence shown in SEQ ID NO: 24 or a sequence that is one nucleotide different from sequence A, antisense The nucleotide sequence of the chain is the reverse complementary sequence B consisting of 21 consecutive nucleotides in the sequence shown in SEQ ID NO:24 or a sequence that differs by one nucleotide from sequence B. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:24所示序列中23个连续核苷酸组成的序列A或与序列A存在一个核苷酸差异的序列，反义链的核苷酸序列为SEQ ID NO:24所示序列中23个连续核苷酸组成的序列反向互补的序列B或与序列B存在一个核苷酸差异的序列。并且，所述正义链与所述反义链的3’末端均具有2个延伸出所述双链区的突出的核苷酸。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 23 consecutive nucleotides in the sequence shown in SEQ ID NO: 24 or a sequence that is one nucleotide different from sequence A, antisense The nucleotide sequence of the chain is the reverse complementary sequence B consisting of 23 consecutive nucleotides in the sequence shown in SEQ ID NO:24 or a sequence that differs by one nucleotide from sequence B. Furthermore, the 3' ends of both the sense strand and the antisense strand have two protruding nucleotides extending out of the double-stranded region.
示例性的，双链核糖核酸为表1或表1-1中示出的siRNA77～siRNA86、siRNA94～siRNA95、siRNA140～siRNA162、siRNA206～siRNA207中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA77 to siRNA86, siRNA94 to siRNA95, siRNA140 to siRNA162, and siRNA206 to siRNA207 shown in Table 1 or Table 1-1.
在一些具体的实施方式中，正义链的核苷酸序列为SEQ ID NO:25所示序列中19个连续核苷酸组成的序列A，反义链的核苷酸序列为SEQ ID NO:25所示序列中19个连续核苷酸组成的序列反向互补的序列B。并且，所述正义链与所述反义链的3’末端均形成平末端。In some specific embodiments, the nucleotide sequence of the sense strand is sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:25, and the nucleotide sequence of the antisense strand is SEQ ID NO:25 The reverse complementary sequence B consisting of 19 consecutive nucleotides in the sequence shown. Furthermore, the 3' ends of the sense strand and the antisense strand both form blunt ends.
示例性的，双链核糖核酸为表1-1中示出的siRNA129。Exemplarily, the double-stranded ribonucleic acid is siRNA129 shown in Table 1-1.
在一些具体的实施方式中，所述正义链包含如SEQ ID NO:26～120、SEQ ID NO:258～267、SEQ ID NO:298～320、SEQ ID NO:385～386任一项所示的核苷酸序列，所述反义链包含如SEQ ID NO:121～215、SEQ ID NO:268～277、SEQ ID NO:321～343、SEQ ID NO:387～388任一项所示的核苷酸序列。In some specific embodiments, the sense strand includes any one of SEQ ID NO: 26 to 120, SEQ ID NO: 258 to 267, SEQ ID NO: 298 to 320, and SEQ ID NO: 385 to 386. The nucleotide sequence, the antisense strand includes as shown in any one of SEQ ID NO:121～215, SEQ ID NO:268～277, SEQ ID NO:321～343, SEQ ID NO:387～388 Nucleotide sequence.
在一些具体的实施方案中，双链核糖核酸选自如表1或表1-1中所示的任一siRNA。本公开提供的siRNA，其与靶mRNA(ANGPTL3mRNA)结合的特异性高，具有较好的靶mRNA的沉默活性，可以显著抑制ANGPTL3基因表达，用于治疗包括脂质代谢异常在内的ANGPTL3相关的疾病。In some specific embodiments, the double-stranded ribonucleic acid is selected from any siRNA as shown in Table 1 or Table 1-1. The siRNA provided by the present disclosure has high binding specificity to the target mRNA (ANGPTL3 mRNA), has good silencing activity of the target mRNA, can significantly inhibit ANGPTL3 gene expression, and is used to treat ANGPTL3-related diseases including abnormal lipid metabolism. disease.
在一些实施方案中，本公开提供了一种siRNA组合物，其包含表1或表1-1中所示siRNA中的任意一种或两种以上的组合。In some embodiments, the present disclosure provides an siRNA composition comprising any one or a combination of two or more of the siRNAs shown in Table 1 or Table 1-1.
在一些实施方案中，正义链的每个核苷酸彼此独立地为修饰的核苷酸或未修饰的核苷酸。在一些实施方案中，反义链的每个核苷酸彼此独立地为修饰的核苷酸或未修饰的核苷酸。In some embodiments, each nucleotide of the sense strand is independently a modified nucleotide or an unmodified nucleotide. In some embodiments, each nucleotide of the antisense strand is independently a modified nucleotide or an unmodified nucleotide.
在一些实施方案中，正义链中任意相连的两个核苷酸由磷酸二酯键或硫代磷酸二酯键连接。在一些实施方案中，反义链中任意相连的两个核苷酸由磷酸二酯键或硫代磷酸二酯键连接。In some embodiments, any two adjacent nucleotides in the sense strand are connected by a phosphodiester bond or a phosphorothioate diester bond. In some embodiments, any two adjacent nucleotides in the antisense strand are connected by a phosphodiester bond or a phosphorothioate diester bond.
在一些实施方式中，正义链的5’末端核苷酸连接5’磷酸基团或5’磷酸衍生基团。在 一些实施方式中，所述反义链的5’末端核苷酸连接5’磷酸基团或5’磷酸衍生基团。In some embodiments, the 5' terminal nucleotide of the sense strand is linked to a 5' phosphate group or a 5' phosphate derivative group. exist In some embodiments, the 5' terminal nucleotide of the antisense strand is linked to a 5' phosphate group or a 5' phosphate derivative group.
示例性的，5’磷酸基团的结构为：5’磷酸衍生基团的结构包括但不限于：等。As an example, the structure of the 5' phosphate group is: The structures of the 5' phosphate derivative group include but are not limited to: wait.
位于正义链或反义链的5’末端核苷酸连接5’磷酸基团或5’磷酸衍生基团后，形成如下所示结构：After the 5’ terminal nucleotide located in the sense strand or antisense strand is connected to the 5’ phosphate group or 5’ phosphate derivative group, the following structure is formed:
其中，Base表示碱基，例如A、U、G、C或T。R’为羟基或被本领域技术人员所知晓的各类基团所取代，例如，2’-氟代(2’-F)修饰的核苷酸，2’-烷氧基修饰的核苷酸，2’-取代的烷氧基修饰的核苷酸，2’-烷基修饰的核苷酸，2’-取代的烷基修饰的核苷酸，2’-脱氧核糖核苷酸。 Among them, Base represents a base, such as A, U, G, C or T. R' is hydroxyl or substituted by various groups known to those skilled in the art, for example, 2'-fluoro (2'-F) modified nucleotides, 2'-alkoxy modified nucleotides , 2'-substituted alkoxy modified nucleotides, 2'-alkyl modified nucleotides, 2'-substituted alkyl modified nucleotides, 2'-deoxyribonucleotides.
双链核糖核酸修饰物double-stranded ribonucleic acid modifications
本公开的第二方面提供一种双链核糖核酸修饰物。进一步地，双链核糖核酸修饰物为siRNA修饰物。siRNA修饰物在保持较高ANGPTL3mRNA抑制活性的同时，可提高siRNA的稳定性。A second aspect of the present disclosure provides a double-stranded ribonucleic acid modification. Further, the double-stranded ribonucleic acid modification is an siRNA modification. siRNA modifications can improve the stability of siRNA while maintaining high ANGPTL3 mRNA inhibitory activity.
在一些实施方案中，双链核糖核酸修饰物包含至少一个核苷酸的修饰。核苷酸的修饰选自核糖基团的修饰和碱基的修饰中的至少一种。在一些实施方案中，“核苷酸的修饰”是指核苷酸的核糖基团2’位羟基被其他基团取代形成的核苷酸或核苷酸衍生物，或者核苷酸上的碱基是经修饰的碱基的核苷酸。所述核苷酸的修饰不会导致siRNA抑制基因表达的功能明显削弱或丧失。例如，可以选择J.K.Watts,G.F.Deleavey,and M.J.Damha,Chemically modified siRNA:tools and applications.Drug Discov Today,2008,13(19-20):842-55中公开的修饰的核苷酸。通过核苷酸的修饰可以提高siRNA的稳定性，并保持其对ANGPTL3基因的高抑制效率。In some embodiments, double-stranded ribonucleic acid modifications comprise modification of at least one nucleotide. The modification of the nucleotide is at least one selected from the group consisting of modification of the ribose group and modification of the base. In some embodiments, "modification of nucleotides" refers to nucleotides or nucleotide derivatives formed by replacing the 2' hydroxyl group of the ribose group of the nucleotide with other groups, or a base on the nucleotide. A nucleotide whose base is a modified base. The modification of the nucleotide will not cause the siRNA's function of inhibiting gene expression to be significantly weakened or lost. For example, modified nucleotides disclosed in J.K. Watts, G.F. Deleavey, and M.J. Damha, Chemically modified siRNA: tools and applications. Drug Discov Today, 2008, 13(19-20): 842-55, can be selected. The stability of siRNA can be improved through nucleotide modification and its high inhibition efficiency of ANGPTL3 gene can be maintained.
示例性的，修饰的核苷酸具有如下所示结构：Exemplarily, the modified nucleotide has the following structure:
其中，Base表示碱基，例如A、U、G、C或T。核糖基团2’位的羟基被R取代。这些核糖基2’位的羟基可以为本领域技术人员所知晓的各类基团所取代，例如，2’-氟代(2’-F)修饰的核苷酸，2’-烷氧基修饰的核苷酸，2’-取代的烷氧基修饰的核苷酸，2’-烷基修饰的核苷酸，2’-取代的烷基修饰的核苷酸，2’-脱氧核糖核苷酸。 Among them, Base represents a base, such as A, U, G, C or T. The hydroxyl group at the 2' position of the ribose group is substituted with R. The hydroxyl groups at the 2' position of these ribosyl groups can be substituted with various groups known to those skilled in the art, for example, 2'-fluoro (2'-F) modified nucleotides, 2'-alkoxy modified nucleotides nucleotides, 2'-substituted alkoxy modified nucleotides, 2'-alkyl modified nucleotides, 2'-substituted alkyl modified nucleotides, 2'-deoxyribonucleosides acid.
在一些实施方案中，2’-烷氧基修饰的核苷酸为2’-甲氧基(2’-OMe，2’-O-CH3)修饰的核苷酸等等。In some embodiments, the 2'-alkoxy modified nucleotide is a 2'-methoxy (2'-OMe, 2'-O- CH3 ) modified nucleotide, and the like.
在一些实施方案中，2’-取代的烷氧基修饰的核苷酸为2’-甲氧基乙氧基(2’-O-CH2-CH2-O-CH3)修饰的核苷酸，2’-O-CH2-CH＝CH2修饰的核苷酸等。In some embodiments, the 2'-substituted alkoxy modified nucleotide is a 2'-methoxyethoxy (2'-O-CH 2 -CH 2 -O-CH 3 ) modified nucleoside Acid, 2'-O-CH 2 -CH=CH 2 modified nucleotide, etc.
在一些实施方案中，2’-取代的烷基修饰的核苷酸为2’-CH2-CH2-CH＝CH2修饰的核苷酸等等。 In some embodiments, the 2'-substituted alkyl modified nucleotide is a 2'-CH2 - CH2 - CH= CH2 modified nucleotide, and the like.
在一些实施方案中，核苷酸的修饰是碱基的修饰。碱基的修饰可以是本领域技术人员所知晓的各类型修饰。示例性的，碱基的修饰包括但不限于m6A、Ψ、m1A、m5A、ms2i6A、i6A、m3C、m5C、ac4C、m7G、m2 , 2G、m2G、m1G、Q、m5U、mcm5U、ncm5U、ncm5Um、D、mcm5s2U、Inosine(I)、hm5C、s4U、s2U、偶氮苯、Cm、Um、Gm、t6A、yW、ms2t6A或其衍生物。In some embodiments, the modification of the nucleotide is a modification of a base. The modification of the base may be various types of modifications known to those skilled in the art. Exemplary base modifications include but are not limited to m 6 A, Ψ, m 1 A, m 5 A, ms 2 i 6 A, i 6 A, m 3 C, m 5 C, ac 4 C, m 7 G, m 2 , 2 G, m 2 G, m 1 G, Q, m 5 U, mcm 5 U, ncm 5 U, ncm 5 Um, D, mcm 5 s 2 U, Inosine(I), hm 5 C , s 4 U, s 2 U, azobenzene, Cm, Um, Gm, t 6 A, yW, ms 2 t 6 A or its derivatives.
在一些实施方案中，核苷酸衍生物是指能够在核酸中代替核苷酸，但结构不同于腺嘌呤核糖核苷酸、鸟嘌呤核糖核苷酸、胞嘧啶核糖核苷酸、尿嘧啶核糖核苷酸或胸腺嘧啶脱氧核糖核苷酸的化合物。在一些实施方案中，核苷酸衍生物可以是异核苷酸、桥联的核苷酸(bridged nucleic acid，简称BNA)或无环核苷酸。BNA是指受约束的或不能接近的核苷酸。BNA可以含有五元环、六元环、或七元环的具有“固定的”C3’-内切糖缩拢的桥联结构。通常将该桥掺入到该核糖的2’-、4’-位处以提供一个2’,4’-BNA核苷酸，如LNA、ENA、cET等。In some embodiments, a nucleotide derivative refers to a nucleotide that can replace a nucleotide in a nucleic acid, but is structurally different from adenine ribonucleotide, guanine ribonucleotide, cytosine ribonucleotide, uracil ribose Nucleotides or thymine deoxyribonucleotide compounds. In some embodiments, the nucleotide derivative may be an isonucleotide, a bridged nucleic acid (BNA), or acyclic nucleotide. BNA refers to constrained or inaccessible nucleotides. BNA may contain a five-membered ring, a six-membered ring, or a seven-membered ring bridged structure with a "fixed" C3'-endoglycocondensation. The bridge is usually incorporated into the 2'-, 4'-position of the ribose to provide a 2', 4'-BNA nucleotide, such as LNA, ENA, cET, etc.
LNA如式(1)所示，ENA如式(2)所示，cET如式(3)所示：
LNA is shown in formula (1), ENA is shown in formula (2), and cET is shown in formula (3):
无环核苷酸是核苷酸的糖环被打开形成的一类核苷酸，如解锁核酸(UNA)或甘油核酸(GNA)，其中，UNA如式(4)所示，GNA如式(5)所示：
Acyclic nucleotides are a type of nucleotide formed by opening the sugar ring of a nucleotide, such as unlocked nucleic acid (UNA) or glycerol nucleic acid (GNA). UNA is represented by formula (4), and GNA is represented by formula (4). 5) shown:
上述式(4)和式(5)中，R选自H、OH或烷氧基(O-烷基)。In the above formula (4) and formula (5), R is selected from H, OH or alkoxy (O-alkyl).
在一些实施方案中，核苷酸衍生物修饰是指核酸中的核苷酸被替代为核苷酸衍生物。示例性的，核苷酸衍生物选自异核苷酸、LNA、ENA、cET、UNA或GNA。In some embodiments, nucleotide derivative modification refers to a nucleotide in a nucleic acid being replaced with a nucleotide derivative. Exemplarily, the nucleotide derivative is selected from isonucleotides, LNA, ENA, cET, UNA or GNA.
在一些实施方案中，核酸中的核苷酸被替代为异核苷酸，在本公开的上下文中，也称之为异核苷修饰。在一些实施方案中，异核苷修饰包括在欲修饰的siRNA的正义链和/或反义链的一个或多个位点掺入异核苷，以代替天然核苷在相应位置进行偶联。In some embodiments, nucleotides in the nucleic acid are replaced with isonucleotides, also referred to as isonucleoside modifications in the context of this disclosure. In some embodiments, isonucleoside modification includes incorporation of isonucleoside at one or more sites of the sense strand and/or antisense strand of the siRNA to be modified, to replace the natural nucleoside for coupling at the corresponding position.
在一些实施方案中，异核苷修饰采用D-异核苷修饰。在另一些实施方案中，异核苷修饰采用L-异核苷修饰。在又一些实施方案中，异核苷修饰采用D-异核苷修饰和L-异核苷修饰。In some embodiments, the isonucleoside modification employs D-isonucleoside modification. In other embodiments, the isonucleoside modification employs L-isonucleoside modification. In yet other embodiments, the isonucleoside modification employs D-isonucleoside modification and L-isonucleoside modification.
在一些实施方案中，双链核糖核酸修饰物包含至少一个位置处的磷酸二酯键的修饰。在一些实施方式中，磷酸二酯键的修饰是指磷酸二酯键中至少一个氧原子被硫原子取代形成硫代磷酸二酯键。硫代磷酸二酯键可以稳定siRNA的双链结构，保持碱基配对的特异性。示例性的，硫代磷酸二酯键结构如下所示：
In some embodiments, the double-stranded ribonucleic acid modification comprises modification of a phosphodiester bond at at least one position. In some embodiments, the modification of the phosphodiester bond means that at least one oxygen atom in the phosphodiester bond is replaced by a sulfur atom to form a phosphorothioate diester bond. The phosphorothioate diester bond can stabilize the double-stranded structure of siRNA and maintain the specificity of base pairing. An exemplary phosphorothioate diester bond structure is as follows:
在一些实施方案中，双链核糖核酸修饰物包含如下至少一种的化学修饰：In some embodiments, the double-stranded ribonucleic acid modification comprises a chemical modification of at least one of the following:
(1)正义链中至少一个核苷酸的修饰，(1) Modification of at least one nucleotide in the sense strand,
(2)正义链中至少一个位置处的磷酸二酯键的修饰，(2) Modification of the phosphodiester bond at at least one position in the sense strand,
(3)反义链中至少一个核苷酸的修饰，(3) Modification of at least one nucleotide in the antisense strand,
(4)反义链中至少一个位置处的磷酸二酯键的修饰。(4) Modification of the phosphodiester bond at at least one position in the antisense strand.
进一步地，双链核糖核酸修饰物是包含(1)-(4)中至少一种化学修饰的siRNA修饰物。Further, the double-stranded ribonucleic acid modification is an siRNA modification including at least one chemical modification among (1)-(4).
在本公开中，正义链中的序列A与反义链中的序列B互补形成双链区后，序列A与序列B的3’末端可以为如下任意一种所示：In the present disclosure, after sequence A in the sense strand and sequence B in the antisense strand are complementary to form a double-stranded region, the 3’ ends of sequence A and sequence B can be any of the following:
(1)序列A与序列B的3’末端均形成平末端；(1) The 3’ ends of sequence A and sequence B both form blunt ends;
(2)序列A的3’末端具有1-2个延伸出所述双链区的突出的核苷酸，且序列B的3’末端形成平末端；(2) The 3’ end of sequence A has 1-2 protruding nucleotides extending out of the double-stranded region, and the 3’ end of sequence B forms a blunt end;
(3)序列B的3’末端具有1-2个延伸出所述双链区的突出的核苷酸，且序列A的3’末端形成平末端；(3) The 3’ end of sequence B has 1-2 protruding nucleotides extending out of the double-stranded region, and the 3’ end of sequence A forms a blunt end;
(4)序列A的3’末端具有1-2个延伸出所述双链区的突出的核苷酸，且序列B的3’末端具有1-2个延伸出所述双链区的突出的核苷酸。(4) The 3' end of sequence A has 1-2 protruding nucleotides extending out of the double-stranded region, and the 3' end of sequence B has 1-2 protruding nucleotides extending out of the double-stranded region. Nucleotides.
在一些实施方案中，正义链的核苷酸序列为序列A所示的序列，反义链的核苷酸序列为序列B所示的序列。In some embodiments, the nucleotide sequence of the sense strand is the sequence shown in Sequence A, and the nucleotide sequence of the antisense strand is the sequence shown in Sequence B.
在一些实施方案中，当正义链与反义链的核苷酸序列互补形成双链区后，正义链与反义链的3’末端不存在突出的核苷酸时，在正义链和反义链中至少一条链的3’末端添加1-2个核苷酸，作为突出的核苷酸。其中，连接于正义链的3’末端的1-2个核苷酸组成序列D，连接于反义链的3’末端的1-2个核苷酸组成序列E。相应地，正义链的核苷酸序列为序列A连接序列D所示的序列，反义链的核苷酸序列为序列B连接序列E所示的序列。或者，正义链的核苷酸序列为序列A所示的序列，反义链的核苷酸序列为序列B连接序列E所示的序列。或者，正义链的核苷酸序列为序列A连接序列D所示的序列，反义链的核苷酸序列为序列B所示的序列。In some embodiments, when the nucleotide sequences of the sense strand and the antisense strand are complementary to form a double-stranded region, and there are no overhanging nucleotides at the 3' ends of the sense strand and the antisense strand, the sense strand and the antisense strand are Add 1-2 nucleotides to the 3' end of at least one of the strands as overhanging nucleotides. Among them, 1-2 nucleotides connected to the 3' end of the sense strand constitute sequence D, and 1-2 nucleotides connected to the 3' end of the antisense strand constitute sequence E. Correspondingly, the nucleotide sequence of the sense strand is the sequence shown in sequence A connected to sequence D, and the nucleotide sequence of the antisense strand is the sequence shown in sequence B connected to sequence E. Alternatively, the nucleotide sequence of the sense strand is the sequence represented by sequence A, and the nucleotide sequence of the antisense strand is the sequence represented by sequence B connected to sequence E. Alternatively, the nucleotide sequence of the sense strand is the sequence represented by sequence A connected to sequence D, and the nucleotide sequence of the antisense strand is the sequence represented by sequence B.
示例性的，在正义链的3’末端添加2个脱氧核糖核苷酸(TT)作为序列D，在反义链的3’末端添加2个脱氧核糖核苷酸(TT)作为序列E。或者，仅在反义链的3’末端添加2个脱氧核糖核苷酸(TT)作为序列E。或者，仅在正义链的3’末端添加2个脱氧核糖核苷酸(TT)作为序列D。For example, two deoxyribonucleotides (TT) are added to the 3' end of the sense strand as sequence D, and two deoxyribonucleotides (TT) are added to the 3' end of the antisense strand as sequence E. Alternatively, add 2 deoxyribonucleotides (TT) as sequence E only to the 3’ end of the antisense strand. Alternatively, add 2 deoxyribonucleotides (TT) only to the 3’ end of the sense strand as sequence D.
在一些实施方案中，当正义链与反义链的核苷酸序列互补形成双链区后，正义链的3’末端不存在突出的核苷酸时，在正义链的3’末端添加由1-2个核苷酸组成的序列D，作为突出的核苷酸。然后，当序列A连接序列D形成的核苷酸序列在完成化学修饰后，排除由1-2个核苷酸组成的序列D。相应地，在双链核糖核酸修饰物中，正义链的核苷酸序列为序列A所示的序列，反义链的核苷酸序列为序列B所示的序列。或者，在双链核糖核酸修饰物中，正义链的核苷酸序列为序列A所示的序列，反义链的核苷酸序列为序列B连接序列E所示的序列。In some embodiments, when the nucleotide sequences of the sense strand and the antisense strand are complementary to form a double-stranded region, and there is no overhanging nucleotide at the 3' end of the sense strand, 1 is added at the 3' end of the sense strand. -Sequence D consisting of 2 nucleotides as the overhanging nucleotide. Then, when the nucleotide sequence formed by connecting sequence A to sequence D is chemically modified, sequence D consisting of 1-2 nucleotides is excluded. Correspondingly, in the double-stranded ribonucleic acid modification, the nucleotide sequence of the sense strand is the sequence shown in sequence A, and the nucleotide sequence of the antisense strand is the sequence shown in sequence B. Alternatively, in the double-stranded ribonucleic acid modification, the nucleotide sequence of the sense strand is the sequence represented by sequence A, and the nucleotide sequence of the antisense strand is the sequence represented by sequence B connected to sequence E.
在一些实施方案中，当序列A在与序列B互补形成双链区后，序列A的3’末端具有延伸出双链区的突出的1-2个核苷酸时，将序列A中位于3’末端的突出的核苷酸排除后作为正义链的核苷酸序列。排除掉3’末端的突出的核苷酸的序列称为序列A’。相应地，双链核糖核酸修饰物的正义链的核苷酸序列为序列A’所示的序列，双链核糖核酸修饰物的反义链的核苷酸序列为序列B所示的序列。或者，双链核糖核酸修饰物的正义链的核 苷酸序列为序列A’所示的序列，双链核糖核酸修饰物的反义链的核苷酸序列为序列B连接序列E所示的序列。In some embodiments, when sequence A is complementary to sequence B to form a double-stranded region, and the 3' end of sequence A has 1-2 protruding nucleotides extending out of the double-stranded region, the sequence A is located at 3 'The overhanging nucleotide at the end is excluded as the nucleotide sequence of the sense strand. The sequence excluding the overhanging nucleotide at the 3' end is called sequence A'. Correspondingly, the nucleotide sequence of the sense strand of the double-stranded ribonucleic acid modified product is the sequence shown in sequence A', and the nucleotide sequence of the antisense strand of the double-stranded ribonucleic acid modified product is the sequence shown in sequence B. Alternatively, the core of the sense strand of the double-stranded ribonucleic acid modification The nucleotide sequence is the sequence shown in sequence A', and the nucleotide sequence of the antisense strand of the double-stranded ribonucleic acid modification is the sequence shown in sequence B connecting sequence E.
在一些实施方案中，沿5’末端向3’末端方向，siRNA修饰物的正义链包括如下修饰：正义链中第7位、第9位、第10位和第11位的核糖核苷酸为2’-氟代修饰的核糖核苷酸；正义链中其他位置的核糖核苷酸为2’-甲氧基修饰的核糖核苷酸。In some embodiments, along the 5' end to the 3' end direction, the sense strand of the siRNA modification includes the following modifications: the ribonucleotides at positions 7, 9, 10 and 11 in the sense strand are 2'-fluoro modified ribonucleotide; ribonucleotides at other positions in the sense strand are 2'-methoxy modified ribonucleotides.
在一些实施方案中，沿5’末端向3’末端方向，siRNA修饰物的正义链包括如下所示位置处的硫代磷酸二酯键：5’末端起始的第1个核苷酸与第2个核苷酸之间，5’末端起始的第2个核苷酸与第3个核苷酸之间，3’末端起始的第1个核苷酸与第2个核苷酸之间，以及3’末端起始的第2个核苷酸与第3个核苷酸之间。In some embodiments, along the 5' end to the 3' end, the sense strand of the siRNA modification includes a phosphorothioate diester bond at the following position: the 1st nucleotide starting from the 5' end and the 1st nucleotide starting from the 5' end. Between 2 nucleotides, between the 2nd nucleotide starting from the 5' end and the 3rd nucleotide, between the 1st nucleotide starting from the 3' end and the 2nd nucleotide starting from the 3' end between the 2nd and 3rd nucleotides starting from the 3' end.
在一些实施方案中，沿5’末端向3’末端方向，siRNA修饰物的正义链包括如下所示位置处的硫代磷酸二酯键：5’末端起始的第1个核苷酸与第2个核苷酸之间、5’末端起始的第2个核苷酸与第3个核苷酸之间。In some embodiments, along the 5' end to the 3' end, the sense strand of the siRNA modification includes a phosphorothioate diester bond at the following position: the 1st nucleotide starting from the 5' end and the 1st nucleotide starting from the 5' end. Between 2 nucleotides, between the 2nd nucleotide and the 3rd nucleotide starting from the 5' end.
在一些具体的实施方案中，siRNA修饰物正义链具有如(a1)-(a3)任一项所示的结构：In some specific embodiments, the sense strand of the siRNA modification has a structure shown in any one of (a 1 )-(a 3 ):
(a1)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-(s)-T-(s)-T-3’，(a 1 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3'，
(a2)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(a 2 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
(a3)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’；(a 3 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3';
其中，N1-N23彼此独立地选自碱基为A、U、C或G的核糖核苷酸，大写字母T表示碱基为胸腺嘧啶的脱氧核糖核苷酸，小写字母m表示该字母m右侧相邻的一个核糖核苷酸为2’-O-CH3修饰的核糖核苷酸，小写字母f表示该字母f左侧相邻的一个核糖核苷酸为2’-F修饰的核糖核苷酸，-(s)-表示前后相邻的两个核苷酸以硫代磷酸二酯键连接。Among them, N 1 to N 23 are independently selected from ribonucleotides whose bases are A, U, C or G. The capital letter T represents deoxyribonucleotides whose base is thymine, and the lowercase letter m represents this letter. The ribonucleotide adjacent to the right side of m is a 2'-O-CH 3 modified ribonucleotide. The lowercase letter f indicates that the ribonucleotide adjacent to the left side of the letter f is 2'-F modified. Ribonucleotide, -(s)- means that two adjacent nucleotides are connected by a phosphorothioate diester bond.
在另外一些具体的实施方案中，siRNA修饰物正义链具有如(a4)-(a5)任一项所示的结构：In other specific embodiments, the sense strand of the siRNA modification has a structure shown in any one of (a 4 ) to (a 5 ):
(a4)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-3’，(a 4 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -3',
(a5)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-mN20-mN21-3’；(a 5 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -3';
其中，N1-N23彼此独立地选自碱基为A、U、C或G的核糖核苷酸，大写字母T表示碱基为胸腺嘧啶的脱氧核糖核苷酸，小写字母m表示该字母m右侧相邻的一个核糖核苷酸为2’-O-CH3修饰的核糖核苷酸，小写字母f表示该字母f左侧相邻的一个核糖核苷酸为2’-F修饰的核糖核苷酸，-(s)-表示前后相邻的两个核苷酸以硫代磷酸二酯键连接。Among them, N 1 to N 23 are independently selected from ribonucleotides whose bases are A, U, C or G. The capital letter T represents deoxyribonucleotides whose base is thymine, and the lowercase letter m represents this letter. The ribonucleotide adjacent to the right side of m is a 2'-O-CH 3 modified ribonucleotide. The lowercase letter f indicates that the ribonucleotide adjacent to the left side of the letter f is 2'-F modified. Ribonucleotide, -(s)- means that two adjacent nucleotides are connected by a phosphorothioate diester bond.
在一些实施方案中，沿5’末端向3’末端方向，siRNA修饰物的反义链包括如下修饰：所述反义链中任意奇数位置处的核糖核苷酸为2’-甲氧基修饰的核糖核苷酸，所述反义链中任意偶数位置处的核糖核苷酸为2’-氟代修饰的核糖核苷酸。In some embodiments, along the 5' end to the 3' end, the antisense strand of the siRNA modification includes the following modifications: ribonucleotides at any odd-numbered positions in the antisense strand are 2'-methoxy modified The ribonucleotide at any even-numbered position in the antisense strand is a 2'-fluorinated modified ribonucleotide.
在一些实施方案中，沿5’末端向3’末端方向，siRNA修饰物的反义链包括如下修饰：所述反义链中第2位、第6位、第14位和第16位的核糖核苷酸为2’-F修饰的核糖核苷酸，所述反义链中其余位置的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸。In some embodiments, along the 5' end to the 3' end direction, the antisense strand of the siRNA modification includes the following modifications: ribose at positions 2, 6, 14 and 16 in the antisense strand The nucleotides are 2'-F modified ribonucleotides, and the remaining ribonucleotides in the antisense strand are 2'-O- CH modified ribonucleotides.
在一些实施方案中，沿5’末端向3’末端方向，siRNA修饰物的反义链包括如下修饰：所述反义链中第2位、第6位、第8位、第9位、第14位和第16位的核糖核苷酸为2’-F修饰的核糖核苷酸，所述反义链中其余位置的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸。In some embodiments, along the direction from the 5' end to the 3' end, the antisense strand of the siRNA modification includes the following modifications: the 2nd, 6th, 8th, 9th, and 9th positions in the antisense strand. The ribonucleotides at positions 14 and 16 are 2'-F modified ribonucleotides, and the remaining ribonucleotides in the antisense strand are 2'-O-CH 3 modified ribonucleotides. .
在一些实施方案中，沿5’末端向3’末端方向，所述反义链中第2位、第14位和第16位的核糖核苷酸为2’-F修饰的核糖核苷酸，所述反义链中第6位的核糖核苷酸为核苷酸衍生物GNA修饰的核糖核苷酸，所述反义链中其余位置的核糖核苷酸为2’-O-CH3修饰的核糖 核苷酸。In some embodiments, along the 5' end to the 3' end, the ribonucleotides at positions 2, 14 and 16 in the antisense strand are 2'-F modified ribonucleotides, The ribonucleotide at position 6 in the antisense strand is a ribonucleotide modified by the nucleotide derivative GNA, and the ribonucleotides at the remaining positions in the antisense strand are modified with 2'-O-CH 3 of ribose Nucleotides.
在一些实施方案中，沿5’末端向3’末端方向，siRNA修饰物的反义链包括如下修饰：所述反义链中第2位、第6位、第14位和第16位的核糖核苷酸为2’-F修饰的核糖核苷酸，所述反义链中第7位的核糖核苷酸为核苷酸衍生物GNA修饰的核糖核苷酸，所述反义链中其余位置的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸。In some embodiments, along the 5' end to the 3' end direction, the antisense strand of the siRNA modification includes the following modifications: ribose at positions 2, 6, 14 and 16 in the antisense strand The nucleotide is a 2'-F modified ribonucleotide, the ribonucleotide at position 7 in the antisense strand is a ribonucleotide modified by the nucleotide derivative GNA, and the remaining ribonucleotides in the antisense strand The ribonucleotide at position is a 2'-O-CH 3 modified ribonucleotide.
在一些实施方案中，沿5’末端向3’末端方向，siRNA修饰物的反义链包括如下所示位置处的硫代磷酸二酯键：5’末端起始的第1个核苷酸与第2个核苷酸之间，5’末端起始的第2个核苷酸与第3个核苷酸之间，3’末端起始的第1个核苷酸与第2个核苷酸之间，以及3’末端起始的第2个核苷酸与第3个核苷酸之间。In some embodiments, along the 5' end to the 3' end, the antisense strand of the siRNA modification includes a phosphorothioate diester bond at the following position: the 1st nucleotide starting from the 5' end and Between the 2nd nucleotide, between the 2nd nucleotide and the 3rd nucleotide starting from the 5' end, between the 1st nucleotide and the 2nd nucleotide starting from the 3' end between the 2nd and 3rd nucleotides starting from the 3' end.
在一些实施方案中，沿5’末端向3’末端方向，反义链的5’末端的核苷酸连接5’磷酸基团或5’磷酸衍生基团。示例性的，5’磷酸基团的结构为：5’磷酸衍生基团的结构包括但不限于：等。In some embodiments, a 5' phosphate group or a 5' phosphate derivative group is attached to the nucleotide at the 5' end of the antisense strand in the direction from the 5' end to the 3' end. As an example, the structure of the 5' phosphate group is: The structures of the 5' phosphate derivative group include but are not limited to: wait.
在一些具体的实施方案中，siRNA修饰物反义链具有如下(b1)-(b15)任一项所示的结构：In some specific embodiments, the siRNA modification antisense strand has the structure shown in any one of the following (b 1 )-(b 15 ):
(b1)5’-P1mN1-(s)-N2f-(s)-mN3-N4f-mN5-N6f-mN7-N8f-mN9-N10f-mN11-N12f-mN13-N14f-mN15-N16f-mN17-N18f-mN19-(s)-T-(s)-T-3’，(b 1 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -N 4 f-mN 5 -N 6 f-mN 7 -N 8 f-mN 9 -N 10 f- mN 11 -N 12 f-mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -N 18 f-mN 19 -(s)-T-(s)-T-3',
(b2)5’-P1mN1-(s)-N2f-(s)-mN3-N4f-mN5-N6f-mN7-N8f-mN9-N10f-mN11-N12f-mN13-N14f-mN15-N16f-mN17-N18f-mN19-(s)-N20f-(s)-mN21-3’，(b 2 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -N 4 f-mN 5 -N 6 f-mN 7 -N 8 f-mN 9 -N 10 f- mN 11 -N 12 f-mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -N 18 f-mN 19 -(s)-N 20 f-(s)-mN 21 -3',
(b3)5’-P1mN1-(s)-N2f-(s)-mN3-N4f-mN5-N6f-mN7-N8f-mN9-N10f-mN11-N12f-mN13-N14f-mN15-N16f-mN17-N18f-mN19-N20f-mN21-(s)-N22f-(s)-mN23-3’；(b 3 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -N 4 f-mN 5 -N 6 f-mN 7 -N 8 f-mN 9 -N 10 f- mN 11 -N 12 f-mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -N 18 f-mN 19 -N 20 f-mN 21 -(s)-N 22 f-(s)- mN 23 -3';
(b4)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N1 4f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 4 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -mN 8 -mN 9 -mN 10 -mN 11 - mN 12 -mN 13 -N 1 4 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3'，
(b5)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N1 4f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 5 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -mN 8 -mN 9 -mN 10 -mN 11 - mN 12 -mN 13 -N 1 4 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
(b6)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N1 4f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(b 6 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -mN 8 -mN 9 -mN 10 -mN 11 - mN 12 -mN 13 -N 1 4 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3',
(b7)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-N8f-N9f-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 7 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -N 8 fN 9 f-mN 10 -mN 11 - mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3'，
(b8)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-N8f-N9f-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 8 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -N 8 fN 9 f-mN 10 -mN 11 - mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
(b9)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-N8f-N9f-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(b 9 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -N 8 fN 9 f-mN 10 -mN 11 - mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3',
(b10)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-[GNA]N6-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 10 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -[GNA]N 6 -mN 7 -mN 8 -mN 9 -mN 10 -mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3',
(b11)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-[GNA]N6-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 11 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -[GNA]N 6 -mN 7 -mN 8 -mN 9 -mN 10 -mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
(b12)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-[GNA]N6-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(b 12 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -[GNA]N 6 -mN 7 -mN 8 -mN 9 -mN 10 -mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3' ,
(b13)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-[GNA]N7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 13 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-[GNA]N 7 -mN 8 -mN 9 -mN 10 - mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3',
(b14)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-[GNA]N7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’， (b 14 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-[GNA]N 7 -mN 8 -mN 9 -mN 10 - mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
(b15)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-[GNA]N7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’；(b 15 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-[GNA]N 7 -mN 8 -mN 9 -mN 10 - mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3 ';
其中，N1-N23彼此独立地选自碱基为A、U、C或G的核糖核苷酸，大写字母T表示碱基为胸腺嘧啶的脱氧核糖核苷酸，小写字母m表示该字母m右侧相邻的一个核糖核苷酸为2’-O-CH3修饰的核糖核苷酸，小写字母f表示该字母f左侧相邻的一个核糖核苷酸为2’-F修饰的核糖核苷酸，P1表示该字母右侧相邻的一个核苷酸为5’-磷酸核苷酸，-(s)-表示前后相邻的两个核苷酸以硫代磷酸二酯键连接，[GNA]表示其右侧相邻的一个核糖核苷酸为存在GNA修饰的核糖核苷酸。Among them, N 1 to N 23 are independently selected from ribonucleotides whose bases are A, U, C or G. The capital letter T represents deoxyribonucleotides whose base is thymine, and the lowercase letter m represents this letter. The ribonucleotide adjacent to the right side of m is a 2'-O-CH 3 modified ribonucleotide. The lowercase letter f indicates that the ribonucleotide adjacent to the left side of the letter f is 2'-F modified. Ribonucleotide, P1 indicates that the adjacent nucleotide on the right side of the letter is a 5'-phosphate nucleotide, -(s)- indicates that the two adjacent nucleotides are connected by a phosphorothioate diester bond , [GNA] indicates that the ribonucleotide adjacent to the right side is a ribonucleotide modified by GNA.
在一些可选的实施方式中，所述正义链包含如SEQ ID NO:216～236、SEQ ID NO:278～287、SEQ ID NO:344～353、SEQ ID NO:389～393任一项所示的核苷酸序列，并且所述反义链包含如SEQ ID NO:237～254、SEQ ID NO:288～297、SEQ ID NO:354～373、SEQ ID NO:377～378、SEQ ID NO:381～384、SEQ ID NO:394～403任一项所示的核苷酸序列。In some optional embodiments, the sense strand includes any one of SEQ ID NO:216～236, SEQ ID NO:278～287, SEQ ID NO:344～353, SEQ ID NO:389～393. The nucleotide sequence shown, and the antisense strand includes such as SEQ ID NO:237～254, SEQ ID NO:288～297, SEQ ID NO:354～373, SEQ ID NO:377～378, SEQ ID NO :381～384, the nucleotide sequence shown in any one of SEQ ID NO:394～403.
在一些实施方案中，双链核糖核酸修饰物包括但不限于如表2中所示的siRNA修饰物。In some embodiments, double-stranded ribonucleic acid modifications include, but are not limited to, siRNA modifications as shown in Table 2.
双链核糖核酸缀合物Double-stranded ribonucleic acid conjugates
本公开的第三方面提供一种双链核糖核酸缀合物，是本公开第一方面提供的双链核糖核酸或第二方面提供的双链核糖核酸修饰物与缀合基团缀合连接得到。The third aspect of the present disclosure provides a double-stranded ribonucleic acid conjugate, which is obtained by conjugating the double-stranded ribonucleic acid provided by the first aspect of the present disclosure or the double-stranded ribonucleic acid modification provided by the second aspect and a conjugation group. .
在本公开中，双链核糖核酸缀合物的正义链与反义链形成双链核糖核酸缀合物的双链区，并且，在双链核糖核酸缀合物的正义链的3’末端形成平末端。在一些实施方案中，双链核糖核酸缀合物的正义链的3’末端形成平末端，双链核糖核酸缀合物的反义链的3’末端具有1-2个延伸出所述双链区的突出的核苷酸。在另外一些实施方案中，双链核糖核酸缀合物的正义链的3’末端形成平末端，双链核糖核酸缀合物的反义链的3’末端形成平末端。In the present disclosure, the sense strand and the antisense strand of the double-stranded ribonucleic acid conjugate form a double-stranded region of the double-stranded ribonucleic acid conjugate, and are formed at the 3' end of the sense strand of the double-stranded ribonucleic acid conjugate. Flat ends. In some embodiments, the 3' end of the sense strand of the double-stranded ribonucleic acid conjugate forms a blunt end and the 3' end of the antisense strand of the double-stranded ribonucleic acid conjugate has 1-2 extensions out of the duplex. overhanging nucleotides in the region. In other embodiments, the 3' end of the sense strand of the double-stranded ribonucleic acid conjugate forms a blunt end, and the 3' end of the antisense strand of the double-stranded ribonucleic acid conjugate forms a blunt end.
在一些优选地实施方案中，双链核糖核酸缀合物由双链核糖核酸修饰物与缀合基团缀合连接得到。其中，双链核糖核酸修饰物的正义链与反义链互补形成双链核糖核酸修饰物的双链区，并且，双链核糖核酸修饰物的正义链的3’末端形成平末端，缀合基团与具有平末端的正义链的3’末端缀合连接，形成双链核糖核酸缀合物。In some preferred embodiments, the double-stranded ribonucleic acid conjugate is obtained by conjugating a double-stranded ribonucleic acid modification with a conjugating group. Wherein, the sense strand and the antisense strand of the double-stranded ribonucleic acid modification are complementary to form the double-stranded region of the double-stranded ribonucleic acid modification, and the 3' end of the sense strand of the double-stranded ribonucleic acid modification forms a blunt end, and the conjugation group The group is conjugated to the 3' end of the sense strand with a blunt end to form a double-stranded ribonucleic acid conjugate.
示例性地，双链核糖核酸修饰物的正义链为序列A所示的序列，反义链为序列B连接序列E所示的序列。并且，双链核糖核酸修饰物的正义链的3’末端形成平末端，双链核糖核酸修饰物的正义链的3’末端连接缀合基团，形成双链核糖核酸缀合物。For example, the sense strand of the double-stranded ribonucleic acid modification is the sequence shown in sequence A, and the antisense strand is the sequence shown in sequence B connected to sequence E. Furthermore, the 3' end of the sense strand of the double-stranded ribonucleic acid modified product is blunt-ended, and the 3' end of the sense strand of the double-stranded ribonucleic acid modified product is connected to a conjugation group to form a double-stranded ribonucleic acid conjugate.
示例性地，双链核糖核酸修饰物的正义链为序列A所示的序列，反义链为序列B所示的序列。并且，双链核糖核酸修饰物的正义链的3’末端形成平末端，双链核糖核酸修饰物的正义链3’末端连接缀合基团，形成双链核糖核酸缀合物。For example, the sense strand of the double-stranded ribonucleic acid modification is the sequence shown in sequence A, and the antisense strand is the sequence shown in sequence B. Furthermore, the 3' end of the sense strand of the double-stranded ribonucleic acid modified product forms a blunt end, and a conjugation group is connected to the 3' end of the sense strand of the double-stranded ribonucleic acid modified product to form a double-stranded ribonucleic acid conjugate.
示例性地，双链核糖核酸修饰物的正义链为序列A连接序列D所示的序列，反义链为序列B连接序列E所示的序列。并且，双链核糖核酸修饰物的正义链的3’末端具有突出的1-2个核苷酸组成的序列D，将双链核糖核酸修饰物中正义链的3’末端的序列D排除后，在序列A的3’末端连接缀合基团，形成双链核糖核酸缀合物。For example, the sense strand of the double-stranded ribonucleic acid modification is the sequence shown in sequence A connected to sequence D, and the antisense strand is the sequence shown in sequence B connected to sequence E. Moreover, the 3' end of the sense strand of the double-stranded ribonucleic acid modified product has a protruding sequence D consisting of 1-2 nucleotides. After excluding the sequence D at the 3' end of the sense strand of the double-stranded ribonucleic acid modified product, A conjugation group is connected to the 3' end of sequence A to form a double-stranded ribonucleic acid conjugate.
示例性地，双链核糖核酸修饰物的正义链为序列A连接序列D所示的序列，反义链为序列B所示的序列。并且，双链核糖核酸修饰物的正义链的3’末端具有突出的1-2个核苷酸组成的序列D，将双链核糖核酸修饰物中正义链的3’末端的序列D排除后，在序列A的3’末端连接缀合基团，形成双链核糖核酸缀合物。For example, the sense strand of the double-stranded ribonucleic acid modification is the sequence shown in sequence A connected to sequence D, and the antisense strand is the sequence shown in sequence B. Moreover, the 3' end of the sense strand of the double-stranded ribonucleic acid modified product has a protruding sequence D consisting of 1-2 nucleotides. After excluding the sequence D at the 3' end of the sense strand of the double-stranded ribonucleic acid modified product, A conjugation group is connected to the 3' end of sequence A to form a double-stranded ribonucleic acid conjugate.
示例性地，双链核糖核酸修饰物的正义链为序列A所示的序列，反义链为序列B连接序列E所示的序列。其中，序列A的3’末端具有延伸出双链区的突出的核苷酸，将位于序列A中3’末端的突出的核苷酸排除后的序列(又称，序列A’)作为用于连接缀合基团的核苷酸序列。因此，双链核糖核酸缀合物的正义链的核苷酸序列为序列A’所示的序 列，反义链的核苷酸序列为序列B连接序列E所示的序列。For example, the sense strand of the double-stranded ribonucleic acid modification is the sequence shown in sequence A, and the antisense strand is the sequence shown in sequence B connected to sequence E. Among them, the 3' end of sequence A has a protruding nucleotide extending out of the double-stranded region, and the sequence after excluding the protruding nucleotide located at the 3' end of sequence A (also called sequence A') is used. The nucleotide sequence to which the conjugation group is attached. Therefore, the nucleotide sequence of the sense strand of the double-stranded ribonucleic acid conjugate is the sequence shown in sequence A' column, the nucleotide sequence of the antisense strand is the sequence shown by sequence B connected to sequence E.
示例性地，双链核糖核酸修饰物的正义链为序列A所示的序列，反义链为序列B所示的序列。其中，序列A的3’末端具有延伸出双链区的突出的核苷酸，将位于序列A中3’末端的突出的核苷酸排除后的序列(又称，序列A’)作为用于连接缀合基团的核苷酸序列。因此，双链核糖核酸缀合物的正义链的核苷酸序列为序列A’所示的序列，反义链的核苷酸序列为序列B所示的序列。For example, the sense strand of the double-stranded ribonucleic acid modification is the sequence shown in sequence A, and the antisense strand is the sequence shown in sequence B. Among them, the 3' end of sequence A has a protruding nucleotide extending out of the double-stranded region, and the sequence after excluding the protruding nucleotide located at the 3' end of sequence A (also called sequence A') is used. The nucleotide sequence to which the conjugation group is attached. Therefore, the nucleotide sequence of the sense strand of the double-stranded ribonucleic acid conjugate is the sequence shown in sequence A', and the nucleotide sequence of the antisense strand is the sequence shown in sequence B.
在一些可选地实施方案中，双链核糖核酸缀合物的正义链具有如(d1)-(d2)任一项所示的结构：In some alternative embodiments, the sense strand of the double-stranded ribonucleic acid conjugate has a structure shown in any one of (d 1 )-(d 2 ):
(d1)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-L96-3’，(d 1 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -L96-3'，
(d2)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-mN20-mN21-L96-3’；(d 2 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -L96-3';
其中，N1-N23彼此独立地选自碱基为A、U、C或G的核糖核苷酸，大写字母T表示碱基为胸腺嘧啶的脱氧核糖核苷酸，小写字母m表示该字母m右侧相邻的一个核糖核苷酸为2’-O-CH3修饰的核糖核苷酸，小写字母f表示该字母f左侧相邻的一个核糖核苷酸为2’-F修饰的核糖核苷酸，-(s)-表示前后相邻的两个核苷酸以硫代磷酸二酯键连接。L96也即式I所示的缀合物基团GalNAc。Among them, N 1 to N 23 are independently selected from ribonucleotides whose bases are A, U, C or G. The capital letter T represents deoxyribonucleotides whose base is thymine, and the lowercase letter m represents this letter. The ribonucleotide adjacent to the right side of m is a 2'-O-CH 3 modified ribonucleotide. The lowercase letter f indicates that the ribonucleotide adjacent to the left side of the letter f is 2'-F modified. Ribonucleotide, -(s)- means that two adjacent nucleotides are connected by a phosphorothioate diester bond. L96 is also the conjugate group GalNAc shown in formula I.
在一些可选地实施方案中，双链核糖核酸缀合物的反义链具有如(b1)-(b15)任一项所示的结构：In some alternative embodiments, the antisense strand of the double-stranded ribonucleic acid conjugate has a structure shown in any one of (b 1 )-(b 15 ):
(b1)5’-P1mN1-(s)-N2f-(s)-mN3-N4f-mN5-N6f-mN7-N8f-mN9-N10f-mN11-N12f-mN13-N14f-mN15-N16f-mN17-N18f-mN19-(s)-T-(s)-T-3’，(b 1 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -N 4 f-mN 5 -N 6 f-mN 7 -N 8 f-mN 9 -N 10 f- mN 11 -N 12 f-mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -N 18 f-mN 19 -(s)-T-(s)-T-3',
(b2)5’-P1mN1-(s)-N2f-(s)-mN3-N4f-mN5-N6f-mN7-N8f-mN9-N10f-mN11-N12f-mN13-N14f-mN15-N16f-mN17-N18f-mN19-(s)-N20f-(s)-mN21-3’，(b 2 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -N 4 f-mN 5 -N 6 f-mN 7 -N 8 f-mN 9 -N 10 f- mN 11 -N 12 f-mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -N 18 f-mN 19 -(s)-N 20 f-(s)-mN 21 -3',
(b3)5’-P1mN1-(s)-N2f-(s)-mN3-N4f-mN5-N6f-mN7-N8f-mN9-N10f-mN11-N12f-mN13-N14f-mN15-N16f-mN17-N18f-mN19-N20f-mN21-(s)-N22f-(s)-mN23-3’，(b 3 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -N 4 f-mN 5 -N 6 f-mN 7 -N 8 f-mN 9 -N 10 f- mN 11 -N 12 f-mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -N 18 f-mN 19 -N 20 f-mN 21 -(s)-N 22 f-(s)- mN 23 -3',
(b4)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N1 4f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 4 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -mN 8 -mN 9 -mN 10 -mN 11 - mN 12 -mN 13 -N 1 4 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3'，
(b5)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N1 4f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 5 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -mN 8 -mN 9 -mN 10 -mN 11 - mN 12 -mN 13 -N 1 4 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
(b6)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N1 4f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(b 6 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -mN 8 -mN 9 -mN 10 -mN 11 - mN 12 -mN 13 -N 1 4 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3',
(b7)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-N8f-N9f-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 7 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -N 8 fN 9 f-mN 10 -mN 11 - mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3'，
(b8)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-N8f-N9f-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 8 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -N 8 fN 9 f-mN 10 -mN 11 - mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
(b9)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-N8f-N9f-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(b 9 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -N 8 fN 9 f-mN 10 -mN 11 - mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3',
(b10)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-[GNA]N6-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 10 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -[GNA]N 6 -mN 7 -mN 8 -mN 9 -mN 10 -mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3',
(b11)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-[GNA]N6-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 11 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -[GNA]N 6 -mN 7 -mN 8 -mN 9 -mN 10 -mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
(b12)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-[GNA]N6-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(b 12 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -[GNA]N 6 -mN 7 -mN 8 -mN 9 -mN 10 -mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3' ,
(b13)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-[GNA]N7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’， (b 13 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-[GNA]N 7 -mN 8 -mN 9 -mN 10 - mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3',
(b14)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-[GNA]N7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 14 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-[GNA]N 7 -mN 8 -mN 9 -mN 10 - mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
(b15)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-[GNA]N7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’；(b 15 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-[GNA]N 7 -mN 8 -mN 9 -mN 10 - mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3 ';
其中，N1-N23彼此独立地选自碱基为A、U、C或G的核糖核苷酸，大写字母T表示碱基为胸腺嘧啶的脱氧核糖核苷酸，小写字母m表示该字母m右侧相邻的一个核糖核苷酸为2’-O-CH3修饰的核糖核苷酸，小写字母f表示该字母f左侧相邻的一个核糖核苷酸为2’-F修饰的核糖核苷酸，P1表示该字母右侧相邻的一个核苷酸为5’-磷酸核苷酸，-(s)-表示前后相邻的两个核苷酸以硫代磷酸二酯键连接，[GNA]表示其右侧相邻的一个核糖核苷酸为存在GNA修饰的核糖核苷酸。Among them, N 1 to N 23 are independently selected from ribonucleotides whose bases are A, U, C or G. The capital letter T represents deoxyribonucleotides whose base is thymine, and the lowercase letter m represents this letter. The ribonucleotide adjacent to the right side of m is a 2'-O-CH 3 modified ribonucleotide. The lowercase letter f indicates that the ribonucleotide adjacent to the left side of the letter f is 2'-F modified. Ribonucleotide, P1 indicates that the adjacent nucleotide on the right side of the letter is a 5'-phosphate nucleotide, -(s)- indicates that the two adjacent nucleotides are connected by a phosphorothioate diester bond , [GNA] indicates that the ribonucleotide adjacent to the right side is a ribonucleotide modified by GNA.
进一步地，双链核糖核酸缀合物为siRNA缀合物，其中siRNA缀合物中与缀合基团连接的siRNA分子可以是未修饰的siRNA，或siRNA修饰物。缀合基团修饰的siRNA分子在保持了较高的抑制活性和稳定性的同时，还具有较好的组织、器官靶向性和促进细胞内吞的能力，可降低对其他组织或器官的影响以及减少siRNA分子使用量，可达到减轻毒性和降低成本的目的。可选地，选择表1或表1-1或表2示出的任意一种siRNA分子与缀合基团连接，得到双链核糖核酸缀合物。Further, the double-stranded ribonucleic acid conjugate is an siRNA conjugate, wherein the siRNA molecule connected to the conjugation group in the siRNA conjugate can be unmodified siRNA or siRNA modification. While maintaining high inhibitory activity and stability, siRNA molecules modified with conjugation groups also have better tissue and organ targeting and the ability to promote cell endocytosis, which can reduce the impact on other tissues or organs. As well as reducing the amount of siRNA molecules used, the purpose of reducing toxicity and reducing costs can be achieved. Alternatively, select any siRNA molecule shown in Table 1 or Table 1-1 or Table 2 and connect it to a conjugation group to obtain a double-stranded ribonucleic acid conjugate.
siRNA与缀合基团的缀合位点可以在siRNA正义链的3’末端或5’末端，也可在反义链的5’端，还可以在siRNA的内部序列中。在一些实施方案中，所述siRNA与缀合基团的缀合位点在siRNA正义链的3’末端。The conjugation site of siRNA and the conjugation group can be at the 3’ end or 5’ end of the sense strand of siRNA, or at the 5’ end of the antisense strand, or in the internal sequence of siRNA. In some embodiments, the conjugation site of the siRNA and the conjugation group is at the 3' end of the sense strand of the siRNA.
在一些实施方式中，所述缀合基团可以连接在核苷酸的磷酸基团、2’-位羟基或者碱基上。在一些实施方式中，所述缀合基团还可以连接在3’-位羟基上，此时核苷酸之间采用2’,5’-磷酸二酯键连接。当缀合基团连接在siRNA链的末端时，所述缀合基团通常连接在核苷酸的磷酸基团上；当缀合基团连接在siRNA的内部序列时，所述缀合基团通常连接在核糖糖环或者碱基上。各种连接方式可以参考文献：Muthiah Manoharan et.al.siRNA conjugates carrying sequentially assembled trivalent N-acetylgalactosamine linked through nucleosides elicit robust gene silencing in vivo in hepatocytes.ACS Chemical biology,2015,10(5):1181-7。In some embodiments, the conjugation group can be attached to the phosphate group, the 2'-hydroxyl group, or the base of the nucleotide. In some embodiments, the conjugation group can also be connected to the 3'-position hydroxyl group, in which case the nucleotides are connected via a 2',5'-phosphodiester bond. When the conjugation group is attached to the end of the siRNA chain, the conjugation group is usually attached to the phosphate group of the nucleotide; when the conjugation group is attached to the internal sequence of the siRNA, the conjugation group Usually attached to the ribose sugar ring or base. For various connection methods, please refer to the literature: Muthiah Manoharan et.al.siRNA conjugates carrying sequentially assembled trivalent N-acetylgalactosamine linked through nucleosides elicit robust gene silencing in vivo in hepatocytes.ACS Chemical Biology, 2015,10(5):1181-7.
在本公开中，缀合基团可以是siRNA给药领域常规使用的配体。在一些实施方式中，所述缀合基团可以选自以下靶向分子或其衍生物形成的配体中的一种或多种：亲脂分子，例如胆固醇、胆汁酸、维生素(例如维生素E)、不同链长的脂质分子；聚合物，例如聚乙二醇；多肽，例如透膜肽；适配体；抗体；量子点；糖类，例如乳糖、聚乳糖、甘露糖、半乳糖、N-乙酰半乳糖胺(GalNAc)；叶酸(folate)；肝实质细胞表达的受体配体，例如去唾液酸糖蛋白、去唾液酸糖残基、脂蛋白(如高密度脂蛋白、低密度脂蛋白等)、胰高血糖素、神经递质(如肾上腺素)、生长因子、转铁蛋白等。In the present disclosure, the conjugating group may be a ligand commonly used in the field of siRNA delivery. In some embodiments, the conjugation group can be selected from one or more ligands formed by the following targeting molecules or derivatives thereof: lipophilic molecules, such as cholesterol, bile acids, vitamins (such as vitamin E ), lipid molecules of different chain lengths; polymers, such as polyethylene glycol; peptides, such as membrane-penetrating peptides; aptamers; antibodies; quantum dots; sugars, such as lactose, polylactose, mannose, galactose, N-acetylgalactosamine (GalNAc); folate; receptor ligands expressed by liver parenchymal cells, such as asialoglycoprotein, asialoglycoside residues, lipoproteins (such as high-density lipoprotein, low-density lipoprotein) Lipoproteins, etc.), glucagon, neurotransmitters (such as epinephrine), growth factors, transferrin, etc.
在一些具体的实施方案中，所述缀合基团具有如下所示结构：
In some specific embodiments, the conjugation group has the structure shown below:
式I所示的缀合基团为GalNAc，GalNAc具有肝脏靶向性，可以将siRNA分子高 特异性地递送于肝脏组织中，特异性抑制肝脏内ANGPTL3基因的高表达。The conjugation group shown in Formula I is GalNAc. GalNAc has liver targeting properties and can bind siRNA molecules to high levels. It is specifically delivered to liver tissue and specifically inhibits the high expression of ANGPTL3 gene in the liver.
在一些具体的实施方案中，GalNAc通过磷酸二酯键与正义链的3’末端缀合连接，得到如下所示结构的siRNA缀合物：
In some specific embodiments, GalNAc is conjugated to the 3' end of the sense strand through a phosphodiester bond, resulting in an siRNA conjugate with the following structure:
其中，双螺旋结构为未修饰的siRNA或siRNA修饰物。Among them, the double helix structure is unmodified siRNA or siRNA modification.
在一些实施方案中，双链核糖核酸缀合物包括但不限于如表3中所示的siRNA缀合物。In some embodiments, double-stranded ribonucleic acid conjugates include, but are not limited to, siRNA conjugates as shown in Table 3.
药物组合物pharmaceutical composition
本公开的第四方面提供一种药物组合物，包括第一方面所述的双链核糖核酸、第二方面所述的双链核糖核酸修饰物，以及第三方面所述的双链核糖核酸缀合物中的一种或多种。The fourth aspect of the present disclosure provides a pharmaceutical composition, including the double-stranded ribonucleic acid described in the first aspect, the double-stranded ribonucleic acid modification described in the second aspect, and the double-stranded ribonucleic acid conjugate described in the third aspect. one or more of the compounds.
在一些实施方案中，所述药物组合物含有如上所述的siRNA作为活性成分和药学上可接受的载体。在本公开中，使用药物组合物的目的在于促进针对生物体的给药，有利于活性成分的吸收，进而发挥生物活性。本公开的药物组合物可以通过任何形式给药，包括注射(动脉内、静脉内、肌肉内、腹膜内、皮下)、粘膜、口服(口服固体制剂、口服液体制剂)、直肠、吸入、植入、局部(例如眼部)给药等。口服固体制剂的非限制性实例包括但不限于散剂、胶囊剂、锭剂、颗粒剂、片剂等。口服或粘膜给药的液体制剂的非限制性实例包括但不限于混悬剂、酊剂、酏剂、溶液剂等。局部给药制剂的非限制性实例包括但不限于乳剂、凝胶剂、软膏剂、乳膏剂、贴剂、糊剂、泡沫剂、洗剂、滴剂或血清制剂。胃肠外给药制剂的非限制性实例包括但不限于注射用溶液剂、注射用干粉剂、注射用悬浮液、注射用乳剂等。本公开的药物组合物还可以制成控制释放或延迟释放剂型(例如脂质体或微球)。In some embodiments, the pharmaceutical composition contains siRNA as described above as an active ingredient and a pharmaceutically acceptable carrier. In the present disclosure, the purpose of using pharmaceutical compositions is to promote administration to living organisms, facilitate the absorption of active ingredients, and thereby exert biological activity. The pharmaceutical compositions of the present disclosure can be administered in any form, including injection (intra-arterial, intravenous, intramuscular, intraperitoneal, subcutaneous), mucosal, oral (oral solid formulation, oral liquid formulation), rectal, inhalation, implantation , local (such as eye) administration, etc. Non-limiting examples of oral solid preparations include, but are not limited to, powders, capsules, lozenges, granules, tablets, and the like. Non-limiting examples of liquid formulations for oral or mucosal administration include, but are not limited to, suspensions, tinctures, elixirs, solutions, and the like. Non-limiting examples of topical formulations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops, or serum formulations. Non-limiting examples of parenteral formulations include, but are not limited to, solutions for injection, dry powders for injection, suspensions for injection, emulsions for injection, and the like. The pharmaceutical compositions of the present disclosure may also be formulated into controlled release or delayed release dosage forms (eg, liposomes or microspheres).
在本公开中，施用途经能够以任何适用的方式进行变化或调整，以满足药物的性质、患者和医务人员的便利以及其它相关因素的需求。In the present disclosure, the methods of administration can be varied or adapted in any applicable manner to meet the needs of the nature of the drug, convenience of the patient and medical staff, and other relevant factors.
医药用途medicinal purposes
本公开的第五方面提供双链核糖核酸，双链核糖核酸修饰物或双链核糖核酸缀合物的如下至少一种用途：A fifth aspect of the present disclosure provides at least one of the following uses of double-stranded ribonucleic acid, double-stranded ribonucleic acid modifications or double-stranded ribonucleic acid conjugates:
(1)抑制ANGPTL3基因表达，或制备用于抑制ANGPTL3基因表达的药物；(1) Inhibit ANGPTL3 gene expression, or prepare drugs for inhibiting ANGPTL3 gene expression;
(2)用于预防或治疗与ANGPTL3基因异常表达相关的疾病，或制备用于预防或治疗与ANGPTL3基因异常表达相关的疾病的药物；(2) For preventing or treating diseases related to abnormal expression of ANGPTL3 gene, or preparing drugs for preventing or treating diseases related to abnormal expression of ANGPTL3 gene;
(3)用于治疗患有将受益于ANGPTL3基因表达降低的疾病的受试者，或制备用于治疗患有将受益于ANGPTL3基因表达降低的疾病的受试者的药物。(3) For use in treating subjects suffering from a disease that would benefit from reduced expression of ANGPTL3 gene, or to prepare a medicament for treating subjects suffering from a disease that would benefit from reduced expression of ANGPTL3 gene.
本公开进一步提供了siRNA分子(包括未修饰的siRNA、siRNA修饰物、siRNA缀合物)或药物组合物在上述(1)-(3)至少一种中的用途。The present disclosure further provides the use of siRNA molecules (including unmodified siRNA, siRNA modifications, siRNA conjugates) or pharmaceutical compositions in at least one of the above (1)-(3).
在本公开中，ANGPTL3基因异常表达，引发如下一种或多种ANGPTL3基因异常表达相关疾病：乳糜微粒血症综合征、2型糖尿病、家族性部分脂肪营养不良、高胆固醇血 症、家族性高胆固醇血症、高甘油三酯血症、非酒精性脂肪肝、动脉粥样硬化、高甘油三酯血症、膝盖损伤及骨关节炎、血脂异常及相关代谢疾病。In the present disclosure, abnormal expression of ANGPTL3 gene causes one or more of the following diseases related to abnormal expression of ANGPTL3 gene: chylomicronemia syndrome, type 2 diabetes, familial partial lipodystrophy, and hypercholesterolemia. disease, familial hypercholesterolemia, hypertriglyceridemia, non-alcoholic fatty liver disease, atherosclerosis, hypertriglyceridemia, knee injuries and osteoarthritis, dyslipidemia and related metabolic diseases.
siRNA分子致使ANGPTL3基因的表达被抑制至少约5％、至少约10％、至少约15％、至少约20％、至少约25％、至少约30％、至少约35％、至少约40％、至少约45％、至少约50％、至少约55％、至少约60％、至少约65％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％、至少约91％、至少约92％、至少约93％、至少约94％、至少约95％、至少约96％、至少约97％、至少约98％或至少约99％，实现对ANGPTL3基因异常表达相关疾病的治疗。The siRNA molecule causes the expression of the ANGPTL3 gene to be inhibited by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least About 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least About 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%, achieve abnormal expression of the ANGPTL3 gene Treatment of related diseases.
在一些实施方案中，本公开提供一种抑制细胞内ANGPTL3基因表达的方法，包括将双链核糖核酸、双链核糖核酸修饰物、双链核糖核酸缀合物或药物组合物与细胞接触。In some embodiments, the present disclosure provides a method of inhibiting ANGPTL3 gene expression in a cell, comprising contacting a double-stranded ribonucleic acid, a double-stranded ribonucleic acid modification, a double-stranded ribonucleic acid conjugate, or a pharmaceutical composition with a cell.
进一步地，抑制细胞内ANGPTL3基因表达的方法，是将siRNA分子(包括未修饰的siRNA、siRNA修饰物、siRNA缀合物)或药物组合物引入细胞内。Further, a method of inhibiting the expression of ANGPTL3 gene in cells is to introduce siRNA molecules (including unmodified siRNA, siRNA modifications, siRNA conjugates) or pharmaceutical compositions into cells.
在一些实施方案中，所述细胞为体内细胞或体外细胞。在一些具体的实施方案中，所述细胞在受试者体内。In some embodiments, the cells are in vivo cells or in vitro cells. In some specific embodiments, the cells are in a subject.
在一些实施方案中，本公开提供预防或治疗疾病的方法，包括向受试者施用双链核糖核酸、双链核糖核酸修饰物、双链核糖核酸缀合物或药物组合物。In some embodiments, the present disclosure provides methods of preventing or treating disease, comprising administering a double-stranded ribonucleic acid, a double-stranded ribonucleic acid modification, a double-stranded ribonucleic acid conjugate, or a pharmaceutical composition to a subject.
进一步地，预防或治疗疾病的方法是向受试者施用siRNA分子(包括未修饰的siRNA、siRNA修饰物、siRNA缀合物)或药物组合物。Further, a method of preventing or treating disease is to administer siRNA molecules (including unmodified siRNA, siRNA modifications, siRNA conjugates) or pharmaceutical compositions to the subject.
在本公开中，“受试者”包括或者人或者非人类动物，优选脊椎动物，并且更优选哺乳动物。受试者可以包括转基因生物体。最优选地，受试者是人。进一步地，受试者具有如下至少一种特性：In this disclosure, "subject" includes either a human or a non-human animal, preferably a vertebrate, and more preferably a mammal. Subjects may include genetically modified organisms. Most preferably, the subject is human. Further, the subject has at least one of the following characteristics:
(1)体内ANGPTL3基因异常表达，更具体地为ANGPTL3基因异常高表达；(1) Abnormal expression of ANGPTL3 gene in vivo, more specifically abnormal high expression of ANGPTL3 gene;
(2)患有与ANGPTL3基因异常表达相关的疾病；(2) Suffering from diseases related to abnormal expression of ANGPTL3 gene;
(3)患有将受益于ANGPTL3基因表达降低的疾病。如罹患或倾向于患上与ANGPTL3基因异常表达相关的疾病的人。(3) Suffering from diseases that would benefit from reduced ANGPTL3 gene expression. Such as people who suffer from or are prone to suffer from diseases related to abnormal expression of ANGPTL3 gene.
表1 siRNA序列信息





Table 1 siRNA sequence information





表1-1 siRNA序列信息

Table 1-1 siRNA sequence information

表2 siRNA修饰物







Table 2 siRNA modifications







上述表格中大写字母“G”、“C”、“A”、“T”和“U”每个通常代表分别含有鸟嘌呤、胞嘧啶、腺嘌呤、胸腺嘧啶和尿嘧啶作为碱基的核苷酸；mA、mU、mC、mG：表示2’-甲氧基修饰的核苷酸；Af、Gf、Cf、Uf：表示2’-氟代修饰的核苷酸；小写字母s表示与该字母s左右相邻的两个核苷酸之间为硫代磷酸酯基连接；P1：表示该P1右侧相邻的一个核苷酸为5’-磷酸核苷酸，[GNA]表示其右侧相邻的一个核糖核苷酸为存在GNA修饰的核糖核苷酸。The capital letters "G", "C", "A", "T" and "U" in the above table each generally represent nucleosides containing guanine, cytosine, adenine, thymine and uracil as bases respectively. Acid; mA, mU, mC, mG: represents 2'-methoxy modified nucleotides; Af, Gf, Cf, Uf: represents 2'-fluoro modified nucleotides; the lowercase letter s represents the same letter The two adjacent nucleotides on the left and right of s are connected by phosphorothioate groups; P1: indicates that the adjacent nucleotide on the right side of P1 is a 5'-phosphate nucleotide, [GNA] indicates its right side The adjacent ribonucleotide is a ribonucleotide modified with GNA.
表3 siRNA缀合物


Table 3 siRNA conjugates


上述表格中大写字母“G”、“C”、“A”、“T”和“U”每个通常代表分别含有鸟嘌呤、胞嘧啶、腺嘌呤、胸腺嘧啶和尿嘧啶作为碱基的核苷酸”；mA、mU、mC、mG：表示2’-甲氧基修饰的核苷酸；Af、Gf、Cf、Uf：表示2’-氟代修饰的核苷酸；小写字母s表示与该字母s左右相邻的两个核苷酸之间为硫代磷酸酯基连接；P1：表示该P1右侧相邻的一个核苷酸为5’-磷酸核苷酸，[GNA]表示其右侧相邻的一个核糖核苷酸为存在GNA 修饰的核糖核苷酸；L96也即式I所示的缀合物基团GalNAc。The capital letters "G", "C", "A", "T" and "U" in the above table each generally represent nucleosides containing guanine, cytosine, adenine, thymine and uracil as bases respectively. "acid"; mA, mU, mC, mG: represent 2'-methoxy modified nucleotides; Af, Gf, Cf, Uf: represent 2'-fluoro modified nucleotides; the lowercase letter s represents the same The two adjacent nucleotides on the left and right of the letter s are connected by phosphorothioate groups; P1: indicates that the adjacent nucleotide on the right side of P1 is a 5'-phosphate nucleotide, and [GNA] indicates its right One ribonucleotide adjacent to the side is GNA Modified ribonucleotide; L96 is also the conjugate group GalNAc shown in formula I.
实施例Example
本公开的其他目的、特征和优点将从以下详细描述中变得明显。但是，应当理解的是，详细描述和具体实施例(虽然表示本公开的具体实施方式)仅为解释性目的而给出，因为在阅读该详细说明后，在本公开的精神和范围内所作出的各种改变和修饰，对于本领域技术人员来说将变得显而易见。Other objects, features and advantages of the present disclosure will become apparent from the following detailed description. It is to be understood, however, that the detailed description and specific examples, while indicating specific embodiments of the disclosure, are given for explanatory purposes only because, after reading the detailed description, reasonable explanations will be made within the spirit and scope of the disclosure. Various changes and modifications will become apparent to those skilled in the art.
本实施例中所用到的实验技术与实验方法，如无特殊说明均为常规技术方法，例如下列实施例中未注明具体条件的实验方法，通常按照常规条件如Sambrook等人，分子克隆：实验室手册(New York:Cold Spring Harbor Laboratory Press,1989)中所述的条件，或按照制造厂商所建议的条件。实施例中所使用的材料、试剂等，如无特殊说明，均可通过正规商业渠道获得。The experimental techniques and experimental methods used in this example are all conventional technical methods unless otherwise specified. For example, the experimental methods without specifying specific conditions in the following examples usually follow conventional conditions, such as Sambrook et al., Molecular Cloning: Experiment The conditions described in the Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or the conditions recommended by the manufacturer. The materials, reagents, etc. used in the examples can be obtained through regular commercial channels unless otherwise specified.
下述实施例涉及的siRNA、siRNA修饰物、siRNA缀合物由天霖生物科技(上海)有限公司合成，实施例中使用的细胞、试剂如表4所示：The siRNA, siRNA modifications, and siRNA conjugates involved in the following examples were synthesized by Tianlin Biotechnology (Shanghai) Co., Ltd. The cells and reagents used in the examples are shown in Table 4:
表4
Table 4
实施例1：siRNA的合成Example 1: Synthesis of siRNA
1.1 siRNA序列设计1.1 siRNA sequence design
根据人ANGPTL3基因mRNA序列，选择不同位点设计多对ANGPTL3siRNAs，设计的所有单个siRNA均能靶向靶基因的所有转录本(如表5)，这些多对siRNA经序列相似性软件比对与其他所有非靶标基因序列有最低同源性。Based on the human ANGPTL3 gene mRNA sequence, multiple pairs of ANGPTL3 siRNAs were designed at different sites. All single siRNAs designed can target all transcripts of the target gene (as shown in Table 5). These multiple pairs of siRNAs were compared with other pairs by sequence similarity software. All non-target gene sequences have minimal homology.
表5
table 5
用于设计siRNA的靶标序列如下所示，靶标序列来源于ANGPTL3的基因mRNA序列(参见NM_014495.4)。The target sequence used to design siRNA is as follows. The target sequence is derived from the gene mRNA sequence of ANGPTL3 (see NM_014495.4).
靶标序列I:
Target sequence I:
靶标序列I-1:
Target sequence I-1:
靶标序列I-2:
Target sequence I-2:
靶标序列I-3:
Target sequence I-3:
靶标序列II:
Target sequence II:
靶标序列II-1:
Target sequence II-1:
靶标序列II-2:
Target sequence II-2:
靶标序列III：
Target sequence III:
靶标序列III-1：
Target sequence III-1:
靶标序列III-2：
Target sequence III-2:
靶标序列IV：
Target sequence IV:
靶标序列IV-1：
Target sequence IV-1:
靶标序列IV-2：
Target sequence IV-2:
靶标序列V：
Target sequence V:
靶标序列V-1：
Target sequence V-1:
靶标序列V-2：
Target sequence V-2:
靶标序列V-3：
Target sequence V-3:
靶标序列VI：
Target sequence VI:
靶标序列VI-1：
Target sequence VI-1:
靶标序列VI-2：
Target sequence VI-2:
靶标序列VI-3：
Target sequence VI-3:
靶标序列VII：
Target sequence VII:
靶标序列VII-1：
Target sequence VII-1:
靶标序列VII-2：
Target sequence VII-2:
靶标序列VII-3：
Target sequence VII-3:
1.2合成方法描述：1.2 Description of synthesis method:
通过固相亚磷酰胺法，按照核苷酸排布顺序自3'-5'方向逐一连接核苷单体。每连接一个核苷单体都包括脱保护、偶联、盖帽、氧化或硫化四步反应。其中，两个核苷酸之间采用磷酸酯连接时，连接后一个核苷单体时，包括脱保护、偶联、盖帽、氧化四步反应。两个核苷酸之间采用硫代磷酸酯连接时，连接后一个核苷单体时，包括保护、偶联、盖帽、硫化四步反应。Through the solid-phase phosphoramidite method, the nucleoside monomers are connected one by one from the 3'-5' direction in the order of nucleotide arrangement. Each connection of a nucleoside monomer involves four steps of deprotection, coupling, capping, oxidation or sulfation. Among them, when two nucleotides are connected using a phosphate ester, when the next nucleoside monomer is connected, it includes four steps of deprotection, coupling, capping, and oxidation. When two nucleotides are connected using phosphorothioate, when the next nucleoside monomer is connected, it includes four steps of protection, coupling, capping and sulfation.
1.3合成条件给定如下：1.3 The synthesis conditions are given as follows:
核苷单体以0.1M浓度的乙腈溶液提供，每一步的脱保护反应的条件相同，即温度为25℃，反应时间为70秒，脱保护试剂为二氯乙酸的二氯甲烷溶液(3％V/V),二氯乙酸与固相载体上4,4’-二甲氧基三苯甲基保护基的摩尔比为5:1。The nucleoside monomer is provided in an acetonitrile solution with a concentration of 0.1M. The conditions for the deprotection reaction in each step are the same, that is, the temperature is 25°C, the reaction time is 70 seconds, and the deprotection reagent is dichloroacetic acid in dichloromethane (3% V/V), the molar ratio of dichloroacetic acid to the 4,4'-dimethoxytrityl protecting group on the solid support is 5:1.
每一步偶联反应条件均相同，包括温度为25℃，固相载体上连接的核酸序列与核苷单体的摩尔比为1:10，固相载体上连接的核酸序列和偶联试剂的摩尔比为1:65，反应时间为600秒，偶联试剂为5-乙硫基-1H-四氮唑的0.5M乙腈溶液。The coupling reaction conditions for each step are the same, including a temperature of 25°C, a molar ratio of the nucleic acid sequence connected to the solid phase carrier to the nucleoside monomer of 1:10, and a molar ratio of the nucleic acid sequence connected to the solid phase carrier to the coupling reagent. The ratio is 1:65, the reaction time is 600 seconds, and the coupling reagent is a 0.5M acetonitrile solution of 5-ethylthio-1H-tetrazole.
每一步盖帽条件均相同，包括温度为25℃，反应时间为15秒。盖帽试剂溶液为摩尔比为1:1的CapA和CapB的混合溶液，盖帽试剂与固相载体上连接的核酸序列的摩尔比为乙酸酐:N-甲基咪唑:固相载体上连接的核酸序列＝1:1:1。The capping conditions in each step are the same, including a temperature of 25°C and a reaction time of 15 seconds. The capping reagent solution is a mixed solution of CapA and CapB with a molar ratio of 1:1. The molar ratio of the capping reagent to the nucleic acid sequence connected to the solid phase carrier is acetic anhydride:N-methylimidazole:the nucleic acid sequence connected to the solid phase carrier. =1:1:1.
每一步氧化反应条件相同，包括温度为25℃，反应时间为15秒，氧化试剂为浓度为0.05M的碘水。碘与偶联步骤中固相载体上连接的核酸序列的摩尔比为30:1。反应在四氢呋喃:水:吡啶＝3:1:1的混合溶剂中进行。The oxidation reaction conditions for each step are the same, including a temperature of 25°C, a reaction time of 15 seconds, and the oxidizing reagent is iodine water with a concentration of 0.05M. The molar ratio of iodine to the nucleic acid sequence attached to the solid support during the coupling step is 30:1. The reaction was carried out in a mixed solvent of tetrahydrofuran:water:pyridine=3:1:1.
每一步硫化反应的条件相同，包括温度为2 5℃，反应时间为300秒，硫化试剂为氢化黄原素。硫化试剂与偶联步骤中固相载体上连接的核酸序列的摩尔比为120:1。反应在乙腈:吡啶＝1:1的混合溶剂中进行。The conditions for each step of the vulcanization reaction are the same, including a temperature of 25°C, a reaction time of 300 seconds, and the vulcanization reagent is hydrogenated xanthogen. The molar ratio of the sulfide reagent to the nucleic acid sequence connected to the solid support in the coupling step is 120:1. The reaction was carried out in a mixed solvent of acetonitrile:pyridine=1:1.
待最后一个核苷单体连接完成后，依次对固相载体上连接的核酸序列进行切割、脱保护、纯化、脱盐，随后冻干得到正义链和反义链；最后将两条链进行加热退火得到产品，冻干，得到冻干粉。After the connection of the last nucleoside monomer is completed, the nucleic acid sequence connected to the solid phase carrier is sequentially cut, deprotected, purified, and desalted, and then freeze-dried to obtain the sense strand and antisense strand; finally, the two strands are heated and annealed. The product is obtained, freeze-dried, and freeze-dried powder is obtained.
实施例2：siRNA缀合物(GalNAc-siRNA)的合成Example 2: Synthesis of siRNA conjugate (GalNAc-siRNA)
2.1 siRNA缀合物具有如下式II所示的结构：
2.1 The siRNA conjugate has the structure shown in Formula II below:
2.2 siRNA缀合物的合成过程2.2 Synthesis process of siRNA conjugates
第一步，通过将DMTr-L96和丁二酸酐反应，得到化合物L96-A：
In the first step, compound L96-A is obtained by reacting DMTr-L96 and succinic anhydride:
制备过程：将DMTr-L96、丁二酸酐、4-二甲基氨基吡啶和二异丙基乙胺加入二氯甲烷中，25℃下搅拌反应24小时，然后用0.5M三乙胺磷酸盐洗涤反应液，水相以二氯甲烷洗涤三次，合并有机相减压蒸干得粗品。然后柱层析纯化得到得到纯品L96-A。Preparation process: Add DMTr-L96, succinic anhydride, 4-dimethylaminopyridine and diisopropylethylamine into dichloromethane, stir and react at 25°C for 24 hours, and then wash with 0.5M triethylamine phosphate. The reaction solution and aqueous phase were washed three times with methylene chloride, and the organic phases were combined and evaporated to dryness under reduced pressure to obtain a crude product. Then column chromatography purified to obtain pure product L96-A.
第二步，将L96-A与NH2-SPS反应得到L96-B：
In the second step, react L96-A with NH 2 -SPS to obtain L96-B:
制备过程：将L96-A、O-苯并三氮唑-四甲基脲六氟磷酸酯(HBTU)和二异丙基乙胺混合溶于乙腈中，室温搅拌5分钟得到均一溶液，加入氨甲基树脂(NH2-SPS，100-200目)至反应液体中，25℃下开始摇床反应，反应18小时后过滤，滤饼依次用二氯甲烷和乙腈洗涤，得滤饼。所得滤饼用CapA/CapB混合溶液进行盖帽反应得到L96-B，即为含有缀合分子的固相载体，然后在偶联反应下将核苷单体连接至缀合分子，随后按照前文所述的siRNA分子合成方法合成连接至缀合物分子的siRNA正义链，采用前文所述的siRNA分子合成方法合成siRNA反义链，退火生成本申请的siRNA缀合物。Preparation process: Mix L96-A, O-benzotriazole-tetramethylurea hexafluorophosphate (HBTU) and diisopropylethylamine in acetonitrile, stir at room temperature for 5 minutes to obtain a uniform solution, add ammonia Add methyl resin (NH 2 -SPS, 100-200 mesh) to the reaction liquid, start shaking reaction at 25°C, filter after 18 hours of reaction, and wash the filter cake with dichloromethane and acetonitrile in sequence to obtain a filter cake. The obtained filter cake is capped with a CapA/CapB mixed solution to obtain L96-B, which is a solid-phase carrier containing the conjugated molecule. Then, the nucleoside monomer is connected to the conjugated molecule under the coupling reaction, and then the nucleoside monomer is connected to the conjugated molecule as described above. The siRNA molecule synthesis method is used to synthesize the siRNA sense strand connected to the conjugate molecule, and the siRNA molecule synthesis method described above is used to synthesize the siRNA antisense strand, and annealed to generate the siRNA conjugate of the present application.
实施例3：siRNA、siRNA修饰物抑制ANGPTL3基因表达Example 3: siRNA and siRNA modifications inhibit ANGPTL3 gene expression
3.1实验材料：3.1 Experimental materials:
Huh7细胞，JCRB cell bank，货号JCRB0403；Huh7 cells, JCRB cell bank, catalog number JCRB0403;
RNA提取试剂盒，96Kit，货号QIAGEN-74182；RNA extraction kit, 96Kit, item number QIAGEN-74182;
转染试剂，购自Invitrogen，货号13778-150； Transfection reagent, purchased from Invitrogen, product number 13778-150;
Opti-medium：减血清培养基，购自Gibco，货号31985-070；Opti-medium: reduced serum medium, purchased from Gibco, Cat. No. 31985-070;
Fastking RT Kit(with gDNase)，购自TianGen，货号KR116-02；Fastking RT Kit (with gDNase), purchased from TianGen, item number KR116-02;
3.2实验方法： 3.2 Experimental methods:
3.2.1取Huh7细胞，先用PBS洗涤后，加入胰蛋白酶进行消化，调整细胞密度至5.5×105细胞/ml，然后以每孔20,000个细胞的密度接种到96孔板中，每孔培养液为100μl。将Huh7细胞置于5％CO2、37℃孵箱中培养过夜。3.2.1 Take Huh7 cells, first wash them with PBS, add trypsin for digestion, adjust the cell density to 5.5×10 5 cells/ml, then seed them into a 96-well plate at a density of 20,000 cells per well, and culture each well The solution is 100μl. Huh7 cells were cultured in 5% CO 2 and 37°C incubator overnight.
3.2.2将待测siRNA及siRNA修饰物(为便于描述，本实施例实验过程描述中统称为siRNA)的干粉以低温高速离心，然后用超纯蒸馏水(ULtraPure Distilled Water)溶解，配制成100μM的siRNA母液。3.2.2 Centrifuge the dry powder of siRNA to be tested and siRNA modifications (for convenience of description, collectively referred to as siRNA in the experimental process description of this example) at low temperature and high speed, then dissolve it in ultrapure distilled water (ULtraPure Distilled Water), and prepare it to 100 μM. siRNA stock solution.
3.2.3配制2nM的siRNA稀释液Z和0.2nM的siRNA稀释液W3.2.3 Prepare 2nM siRNA dilution Z and 0.2nM siRNA dilution W
(1)0.1μM siRNA贮备液X和0.01μM siRNA贮备液Y的制备：(1) Preparation of 0.1μM siRNA stock solution X and 0.01μM siRNA stock solution Y:
a)取上述步骤3.2.2中制得的100μM siRNA母液2μl，加入18μl超纯蒸馏水，得到终浓度为10μM的siRNA稀释液；a) Take 2 μl of the 100 μM siRNA stock solution prepared in step 3.2.2 above, and add 18 μl of ultrapure distilled water to obtain a siRNA dilution with a final concentration of 10 μM;
b)取步骤a)中制得的10μM的siRNA稀释液2μl，加入18μl超纯蒸馏水，得到终浓度为1μM的siRNA稀释液；b) Take 2 μl of the 10 μM siRNA dilution prepared in step a) and add 18 μl of ultrapure distilled water to obtain a final concentration of 1 μM siRNA dilution;
c)取步骤b)中制得的1μM的siRNA稀释液2μl，加入18μl超纯蒸馏水，得到终浓度为0.1μM的siRNA贮备液X；c) Take 2 μl of the 1 μM siRNA dilution prepared in step b) and add 18 μl of ultrapure distilled water to obtain siRNA stock solution X with a final concentration of 0.1 μM;
d)取步骤c)中制得的0.1μM的siRNA贮备液X 2μl，加入18μl超纯蒸馏水，得到终浓度为0.01μM的siRNA贮备液Y；d) Take 2 μl of the 0.1 μM siRNA stock solution X prepared in step c), add 18 μl of ultrapure distilled water, and obtain a siRNA stock solution Y with a final concentration of 0.01 μM;
(2)取上述配置好的siRNA贮备液X和siRNA贮备液Y各2μl，分别加入98μl Opti-medium，分别得到2nM的siRNA稀释液Z和0.2nM的siRNA稀释液W。(2) Take 2 μl each of the above configured siRNA stock solution X and siRNA stock solution Y, and add 98 μl Opti-medium respectively to obtain 2 nM siRNA dilution Z and 0.2 nM siRNA dilution W respectively.
3.2.4转染Huh7细胞3.2.4 Transfection of Huh7 cells
(1)取转染试剂3μl，加入97μl Opti-medium，得到转染试剂稀释液；将转染试剂稀释液与步骤3.2.3中制得的2nM siRNA稀释液Z以1:1体积比混合制备成转染混合物，静置5分钟，取10μl转染混合物加入到96孔板中转染步骤3.2.1中培养的Huh7细胞(终体积100μl，该体系中siRNA的浓度为0.1nM)；(1) take 3μl transfection reagent, add 97μl Opti-medium, get transfection reagent diluent; Mix the transfection reagent dilution and the 2nM siRNA dilution Z prepared in step 3.2.3 at a volume ratio of 1:1 to prepare a transfection mixture. Let it stand for 5 minutes. Add 10 μl of the transfection mixture to a 96-well plate for transfection. Huh7 cells cultured in step 3.2.1 (final volume 100 μl, siRNA concentration in this system is 0.1 nM);
(2)取转染试剂3μl，加入97μl Opti-medium，得到转染试剂稀释液；将转染试剂稀释液与步骤3.2.3中制得的0.2nM siRNA稀释液W以1:1体积比混合制备成转染混合物，静置5分钟，取10μl转染混合物加入到96孔板中转染步骤3.2.1中培养的Huh7细胞(终体积100μl，该体系中siRNA的浓度为0.01nM)。(2) Take 3μl transfection reagent, add 97μl Opti-medium, get transfection reagent diluent; Mix the transfection reagent dilution and the 0.2nM siRNA dilution W prepared in step 3.2.3 at a volume ratio of 1:1 to prepare a transfection mixture. Let it stand for 5 minutes. Add 10 μl of the transfection mixture to a 96-well plate for transfection. Stain the Huh7 cells cultured in step 3.2.1 (the final volume is 100 μl, the concentration of siRNA in this system is 0.01 nM).
上述转染后培养24小时；每个浓度(0.1nM和0.01nM)设置2个重复。Culture for 24 hours after the above transfection; 2 replicates were set for each concentration (0.1 nM and 0.01 nM).
3.2.5根据96Kit试剂盒产品说明书，提取步骤3.2.4中获得的Huh7细胞的总RNA。3.2.5Based on 96Kit kit product instructions, extract the total RNA from Huh7 cells obtained in step 3.2.4.
3.2.6对提取的总RNA使用Fastking RT Kit(with gDNase)进行逆转录至cDNA，按照以下步骤进行：3.2.6 Use Fastking RT Kit (with gDNase) to reverse-transcribe the extracted total RNA to cDNA. Follow the following steps:
a)按照下表用gDNA酶除去gDNA；a) Use gDNAase to remove gDNA according to the table below;
表6
Table 6
42℃，3min；4℃，静置。42℃, 3min; 4℃, let stand.
b)如下所述进行逆转录程序b) Perform the reverse transcription procedure as follows
表7

Table 7

42℃，15min；95℃，3min。42℃, 15min; 95℃, 3min.
c)将逆转录产物储存在-20℃以进行实时PCR分析。c) Store the reverse transcription product at -20°C for real-time PCR analysis.
3.2.7使用FastStart universal probe master(ROX)进行实时PCR分析3.2.7 Real-time PCR analysis using FastStart universal probe master (ROX)
a)如下表所示制备qPCR反应混合物，在整个操作过程中，所有试剂都放置在冰上；a) Prepare the qPCR reaction mixture as shown in the table below, and keep all reagents on ice during the entire operation;
表8
Table 8
b)如下所述进行qPCR程序b) Perform the qPCR procedure as follows
95℃，10分钟；95℃, 10 minutes;
95℃，15秒；60℃，1分钟(40个循环)。95°C, 15 seconds; 60°C, 1 minute (40 cycles).
3.2.8结果分析3.2.8 Result analysis
a)使用Quant Studio 7软件采用默认设置，自动计算Ct值；a) Use Quant Studio 7 software with default settings to automatically calculate the Ct value;
b)使用以下公式计算基因的相对表达量：b) Use the following formula to calculate the relative expression of genes:
ΔCt＝Ct(ANGPTL3基因)–Ct(GAPDH)ΔCt＝Ct(ANGPTL3 gene)–Ct(GAPDH)
ΔΔCt＝ΔCt(检测样品组)-ΔCt(Mock组)ΔΔCt=ΔCt (test sample group)-ΔCt (Mock group)
相对于Mock组的mRNA表达＝2-ΔΔCt，Relative to the mRNA expression of the Mock group=2 -ΔΔCt ,
其中Mock组表示和检测样品组相比，未加入siRNA的组。The Mock group represents the group in which siRNA is not added compared with the test sample group.
抑制率(％)＝(Mock组mRNA相对表达量–检测样品组mRNA相对表达量)/Mock组mRNA相对表达量×100％。Inhibition rate (%) = (relative expression of mRNA in the Mock group – relative expression of mRNA in the test sample group)/relative expression of mRNA in the Mock group × 100%.
3.3沉默实验结果3.3 Silence experiment results
选取浓度0.1nM和0.01nM进行测试。Select concentrations of 0.1nM and 0.01nM for testing.
3.3.1设计序列结果3.3.1 Design sequence results
表9





Table 9





从表9中可以看出，本公开提供的siRNA、siRNA修饰物显示出对ANGPTL3基因的优异的抑制效果。在0.01nM的浓度下，抑制率高达70.49％；在0.1nM的浓度下，抑制率高达86.25％。As can be seen from Table 9, the siRNA and siRNA modifications provided by the present disclosure show excellent inhibitory effects on the ANGPTL3 gene. At a concentration of 0.01nM, the inhibition rate is as high as 70.49%; at a concentration of 0.1nM, the inhibition rate is as high as 86.25%.
3.3.2 IC50实验结果3.3.2 IC 50 experimental results
下述待测siRNA测定浓度范围设置(nM)：10、2.5、0.625、0.156、0.039、0.0098、0.0024、0.0006，再按照与3.2相似的方法进行IC50测定。Set the following siRNA concentration range to be tested (nM): 10, 2.5, 0.625, 0.156, 0.039, 0.0098, 0.0024, 0.0006, and then measure IC 50 according to a method similar to 3.2.
结果分析：Result analysis:
a)使用Quant Studio 7软件采用默认设置，自动计算Ct值；a) Use Quant Studio 7 software with default settings to automatically calculate the Ct value;
b)使用以下公式计算基因的相对表达量：b) Use the following formula to calculate the relative expression of genes:
ΔCt＝Ct(ANGPTL3基因)–Ct(GAPDH)ΔCt＝Ct(ANGPTL3 gene)–Ct(GAPDH)
ΔΔCt＝ΔCt(检测样品组)–ΔCt(Mock组)；ΔΔCt=ΔCt (test sample group) – ΔCt (Mock group);
相对于Mock组的mRNA表达＝2-ΔΔCt Relative to the mRNA expression of the Mock group = 2 -ΔΔCt
抑制率(％)＝(Mock组mRNA相对表达量–检测样品组mRNA相对表达量)/Mock组mRNA相对表达量×100％；Inhibition rate (%) = (relative expression of mRNA in the Mock group – relative expression of mRNA in the test sample group)/relative expression of mRNA in the Mock group × 100%;
其中，Mock组表示和检测样品组相比，未加入siRNA的组。Among them, the Mock group represents the group in which siRNA is not added compared with the test sample group.
以siRNA浓度的log值作为X轴，百分比抑制率为Y轴，采用分析软件GraphPad Prism 8的“log(抑制剂)vs.响应–变量斜率”功能模块，来拟合量效曲线，从而得出各个siRNA的IC50值。Taking the log value of siRNA concentration as the X-axis and the percentage inhibition rate as the Y-axis, use the "log (inhibitor) vs. response-variable slope" function module of the analysis software GraphPad Prism 8 to fit the dose-effect curve, thus obtaining IC50 value of each siRNA.
拟合公式为：Y＝Bottom+(Top–Bottom)/(1+10^((LogIC50–X)×HillSlope))The fitting formula is: Y=Bottom+(Top–Bottom)/(1+10^((LogIC50–X)×HillSlope))
其中：Top表示顶部平台处的百分比抑制率，曲线的Top标准一般在80％至120％；Bottom表示底部平台处的百分比抑制率，曲线的Bottom一般在–20％至20％之间；HillSlope表示百分比抑制率曲线的斜率。Among them: Top represents the percentage inhibition rate at the top platform, and the Top standard of the curve is generally between 80% and 120%; Bottom represents the percentage inhibition rate at the bottom platform, and the Bottom of the curve is generally between –20% and 20%; HillSlope represents The slope of the percent inhibition curve.
表10
Table 10
从表10中可以看出，本公开提供的siRNA修饰物具有优异的ANGPTL3基因抑制 活性，可以在低至0.031nM的IC50时依然具有良好的ANGPTL3基因抑制活性。As can be seen from Table 10, the siRNA modification provided by the present disclosure has excellent ANGPTL3 gene inhibition Activity, it can still have good ANGPTL3 gene inhibitory activity at an IC 50 as low as 0.031nM.
实施例4：递送系统验证Example 4: Delivery System Validation
4.1实验材料：4.1 Experimental materials:
人原代肝细胞PHH细胞，由药明康德提供；Human primary hepatocyte PHH cells were provided by WuXi AppTec;
PHH培养基：invitroGRO CP Meduim serum free BIOVIT，货号：S03316PHH medium: invitroGRO CP Meduim serum free BIOVIT, Cat. No.: S03316
转染试剂，购自Invitrogen，货号：13778-150； Transfection reagent, purchased from Invitrogen, product number: 13778-150;
RNA提取试剂盒96Kit，购自QIAGEN，货号：QIAGEN-74182；RNA extraction kit 96Kit, purchased from QIAGEN, item number: QIAGEN-74182;
逆转录试剂盒FastKing RT Kit(With gDNase)，购自TianGen，货号：KR116-02；Reverse transcription kit FastKing RT Kit (With gDNase), purchased from TianGen, product number: KR116-02;
FastStart Universal Probe master(ROX)，购自Roche，货号：04914058001；FastStart Universal Probe master (ROX), purchased from Roche, item number: 04914058001;
ANGPTL3及GAPDH引物由药明康德提供。ANGPTL3 and GAPDH primers were provided by WuXi AppTec.
4.2实验方法：4.2 Experimental methods:
siRNA缀合物(siRNA缀合物终浓度为10nM、2.5nM、0.63nM、0.16nM、0.04nM、0.01nM、0.0024nM和0.0006nM，复孔)通过转染进入PHH细胞，过程如下所述：取冻存的PHH细胞，复苏，计数，调整细胞到6×105细胞/ml，同时应用RNAiMax转染试剂将siRNA缀合物转入细胞，以每孔54,000个细胞的密度接种到96孔板中，每孔加入PPH培养基100μL。细胞置于5％CO2、37℃孵箱中培养。48小时后，去除培养基并收集细胞用于总RNA提取。根据试剂盒产品说明书使用96Kit提取总RNA。siRNA conjugates (siRNA conjugate final concentrations are 10nM, 2.5nM, 0.63nM, 0.16nM, 0.04nM, 0.01nM, 0.0024nM and 0.0006nM, duplicate wells) into PHH cells through transfection, the process is as follows: Take the frozen PHH cells, resuscitate, count, adjust the cells to 6×10 5 cells/ml, and apply at the same time RNAiMax transfection reagent transfers siRNA conjugates into cells, which are seeded into a 96-well plate at a density of 54,000 cells per well, and 100 μL of PPH medium is added to each well. Cells were cultured in 5% CO 2 and 37°C incubator. After 48 hours, the medium was removed and cells were collected for total RNA extraction. Use according to kit product instructions 96Kit extracted total RNA.
siRNA缀合物(siRNA缀合物终浓度为500nM、125nM、31.25nM、7.81nM、1.95nM、0.49nM、0.12nM和0.03nM，复孔)通过自由摄取进入PHH细胞，过程如下所述：取冻存的PHH细胞，复苏，计数，调整细胞到6×105细胞/ml，同时加入siRNA缀合物，以每孔54,000个细胞的密度接种到96孔板中，每孔培养液为100μl。细胞置于5％CO2、37℃孵箱中培养。48小时后，去除培养基并收集细胞用于总RNA提取。根据试剂盒产品说明书使用96Kit提取总RNA。siRNA conjugates (siRNA conjugate final concentrations are 500nM, 125nM, 31.25nM, 7.81nM, 1.95nM, 0.49nM, 0.12nM and 0.03nM, duplicate wells) enter PHH cells through free uptake. The process is as follows: take Frozen PHH cells were resuscitated, counted, and the cells were adjusted to 6 × 10 5 cells/ml. At the same time, siRNA conjugate was added and seeded into a 96-well plate at a density of 54,000 cells per well, with 100 μl of culture medium per well. Cells were cultured in 5% CO 2 and 37°C incubator. After 48 hours, the medium was removed and cells were collected for total RNA extraction. Use according to kit product instructions 96Kit extracted total RNA.
采用与实施例3中相似的方法，通过逆转录反应将提取的总RNA逆转录为cDNA，以及通过qPCR对逆转录得到的ANGPTL3cDNA进行定量扩增。GAPDH cDNA将作为内部对照进行平行扩增。PCR反应程序为：95℃10分钟，然后进入循环模式，95℃15秒，随后60℃，60秒，共40个循环。Using a method similar to that in Example 3, the extracted total RNA was reverse transcribed into cDNA through reverse transcription reaction, and the ANGPTL3 cDNA obtained by reverse transcription was quantitatively amplified through qPCR. GAPDH cDNA will be amplified in parallel as an internal control. The PCR reaction program is: 95°C for 10 minutes, then enter the cycle mode, 95°C for 15 seconds, then 60°C for 60 seconds, a total of 40 cycles.
4.3结果分析4.3 Result analysis
a)使用Quant Studio 7软件采用默认设置，自动计算Ct值；a) Use Quant Studio 7 software with default settings to automatically calculate the Ct value;
b)使用以下公式计算基因的相对表达量：b) Use the following formula to calculate the relative expression of genes:
ΔCt＝Ct(ANGPTL3基因)–Ct(GAPDH)ΔCt＝Ct(ANGPTL3 gene)–Ct(GAPDH)
ΔΔCt＝ΔCt(检测样品组)-ΔCt(Mock组)ΔΔCt=ΔCt (test sample group)-ΔCt (Mock group)
相对于Mock的mRNA表达＝2-ΔΔCt，Relative to Mock’s mRNA expression=2 -ΔΔCt ,
抑制率(％)＝(Mock组mRNA相对表达量–检测样品组mRNA相对表达量)/Mock组mRNA相对表达量×100％，Inhibition rate (%) = (relative expression of mRNA in the Mock group – relative expression of mRNA in the test sample group)/relative expression of mRNA in the Mock group × 100%,
其中Mock组表示和检测样品组相比，未加入siRNA缀合物的组。The Mock group represents the group in which siRNA conjugate is not added compared with the test sample group.
以siRNA缀合物浓度的log值作为X轴，百分比抑制率为Y轴，采用分析软件GraphPad Prism 8的“log(抑制剂)vs.响应–变量斜率”功能模块，来拟合量效曲线，从而得出各个siRNA缀合物的IC50值。Taking the log value of the siRNA conjugate concentration as the X-axis and the percentage inhibition rate as the Y-axis, use the "log (inhibitor) vs. response-variable slope" function module of the analysis software GraphPad Prism 8 to fit the dose-effect curve. This resulted in the IC50 value for each siRNA conjugate.
拟合公式为：Y＝Bottom+(Top–Bottom)/(1+10^((LogIC50–X)×HillSlope))The fitting formula is: Y=Bottom+(Top–Bottom)/(1+10^((LogIC50–X)×HillSlope))
其中：Top表示顶部平台处的百分比抑制率，曲线的Top标准一般在80％至120％；Bottom表示底部平台处的百分比抑制率，曲线的Bottom一般在–20％至20％之间；HillSlope表示百分比抑制率曲线的斜率。Among them: Top represents the percentage inhibition rate at the top platform, and the Top standard of the curve is generally between 80% and 120%; Bottom represents the percentage inhibition rate at the bottom platform, and the Bottom of the curve is generally between –20% and 20%; HillSlope represents The slope of the percent inhibition curve.
表11
Table 11
上表中的L96也即式I所示的缀合物基团GalNAc。L96 in the above table is also the conjugate group GalNAc shown in formula I.
实施例5：siRNA、siRNA修饰物抑制ANGPTL3基因表达Example 5: siRNA and siRNA modifications inhibit ANGPTL3 gene expression
本实施例中使用的实验材料、实验方法与实施例3中的实验材料、实验方法相同。基因表达沉默实验结果如下表12所示。The experimental materials and experimental methods used in this example are the same as those in Example 3. The results of gene expression silencing experiments are shown in Table 12 below.
表12



Table 12



实施例6：递送系统验证Example 6: Delivery System Validation
本实施例中使用的实验材料、实验方法与实施例4中的实验材料、实验方法相同。IC50实验结果如下表14所示。The experimental materials and experimental methods used in this example are the same as those in Example 4. The IC 50 experimental results are shown in Table 14 below.
表14
Table 14
上表中的L96也即式I所示的缀合物基团GalNAc。L96 in the above table is also the conjugate group GalNAc shown in formula I.
实施例7：siRNA缀合物抑制ANGPTL3基因表达的抑制率测定Example 7: Determination of inhibition rate of siRNA conjugate inhibiting ANGPTL3 gene expression
7.1实验材料：7.1 Experimental materials:
人原代肝细胞PHH细胞，由药明康德提供；Human primary hepatocyte PHH cells were provided by WuXi AppTec;
PHH培养基：invitroGRO CP Meduim serum free BIOVIT，货号：S03316PHH medium: invitroGRO CP Meduim serum free BIOVIT, Cat. No.: S03316
转染试剂，购自Invitrogen，货号：13778-150； Transfection reagent, purchased from Invitrogen, product number: 13778-150;
RNA提取试剂盒96Kit，购自QIAGEN，货号：QIAGEN-74182；RNA extraction kit 96Kit, purchased from QIAGEN, item number: QIAGEN-74182;
逆转录试剂盒FastKing RT Kit(With gDNase)，购自TianGen，货号：KR116-02；Reverse transcription kit FastKing RT Kit (With gDNase), purchased from TianGen, product number: KR116-02;
FastStart Universal Probe master(ROX)，购自Roche，货号：04914058001；FastStart Universal Probe master (ROX), purchased from Roche, item number: 04914058001;
ANGPTL3及GAPDH引物由药明康德提供。ANGPTL3 and GAPDH primers were provided by WuXi AppTec.
7.2实验方法：7.2 Experimental methods:
siRNA缀合物(siRNA缀合物终浓度分别为5nM和0.5nM，复孔)通过转染进入PHH细胞，过程如下所述：取冻存的PHH细胞，复苏，计数，调整细胞到6×105细胞/ml，同时应用转染试剂将siRNA缀合物转入细胞，以每孔54,000个细胞的密度接种到96孔板中，每孔培养液100μL。细胞置于5％CO2、37℃孵箱中培养。48小时后，去除培养基并收集细胞用于总RNA提取。根据试剂盒产品说明书使用96Kit提取总RNA。siRNA conjugates (the final concentrations of siRNA conjugates are 5nM and 0.5nM, respectively, in duplicate wells) are transfected into PHH cells. The process is as follows: take the frozen PHH cells, resuscitate, count, and adjust the cells to 6×10 5 cells/ml, applied simultaneously The transfection reagent transfers the siRNA conjugate into the cells, which are seeded into a 96-well plate at a density of 54,000 cells per well, with 100 μL of culture medium per well. Cells were cultured in 5% CO 2 and 37°C incubator. After 48 hours, the medium was removed and cells were collected for total RNA extraction. Use according to kit product instructions 96Kit extracted total RNA.
siRNA缀合物(siRNA缀合物终浓度分别为100nM和10nM，复孔)通过自由摄取进入PHH细胞，过程如下所述：取冻存的PHH细胞，复苏，计数，调整细胞到6×105 细胞/ml，同时加入siRNA缀合物，以每孔54,000个细胞的密度接种到96孔板中，每孔培养液为100μl。细胞置于5％CO2、37℃孵箱中培养。48小时后，去除培养基并收集细胞用于总RNA提取。根据试剂盒产品说明书使用96Kit提取总RNA。siRNA conjugates (the final concentrations of siRNA conjugates are 100nM and 10nM, respectively, in duplicate wells) enter PHH cells through free uptake. The process is as follows: take the frozen PHH cells, resuscitate, count, and adjust the cells to 6×10 5 cells/ml, and siRNA conjugate was added at the same time, and seeded into a 96-well plate at a density of 54,000 cells per well, with 100 μl of culture medium per well. Cells were cultured in 5% CO 2 and 37°C incubator. After 48 hours, the medium was removed and cells were collected for total RNA extraction. Use according to kit product instructions 96Kit extracted total RNA.
采用与实施例3中相似的方法，通过逆转录反应将提取的总RNA逆转录为cDNA。ANGPTL3cDNA将通过qPCR进行检测。GAPDH cDNA将作为内部对照进行平行检测。PCR反应程序为：95℃10分钟，然后进入循环模式，95℃15秒，随后60℃，60秒，共40个循环。Using a method similar to that in Example 3, the extracted total RNA was reverse transcribed into cDNA through a reverse transcription reaction. ANGPTL3cDNA will be detected by qPCR. GAPDH cDNA will be used as an internal control for parallel testing. The PCR reaction program is: 95°C for 10 minutes, then enter the cycle mode, 95°C for 15 seconds, then 60°C for 60 seconds, a total of 40 cycles.
结果分析：Result analysis:
a)使用Quant Studio 7软件采用默认设置，自动计算Ct值；a) Use Quant Studio 7 software with default settings to automatically calculate the Ct value;
b)使用以下公式计算基因的相对表达量：b) Use the following formula to calculate the relative expression of genes:
ΔCt＝Ct(ANGPTL3基因)–Ct(GAPDH)ΔCt＝Ct(ANGPTL3 gene)–Ct(GAPDH)
ΔΔCt＝ΔCt(检测样品组)–ΔCt(Mock组)，其中Mock组表示和检测样品组相比，未加入siRNA缀合物的组；ΔΔCt=ΔCt (detection sample group) – ΔCt (Mock group), where the Mock group represents the group in which siRNA conjugate is not added compared with the detection sample group;
相对于Mock组的mRNA表达＝2-ΔΔCt Relative to the mRNA expression of the Mock group = 2 -ΔΔCt
抑制率(％)＝(Mock组mRNA相对表达量–检测样品组mRNA相对表达量)/Mock组mRNA相对表达量×100％Inhibition rate (%) = (relative expression of mRNA in the Mock group – relative expression of mRNA in the test sample group)/relative expression of mRNA in the Mock group × 100%
表15


Table 15


从表15中可以看出，本公开提供的siRNA修饰物显示出对ANGPTL3基因的优异的抑制效果。在自由摄取条件下，在100nM浓度的抑制率高达89.37％，在10nM浓度的抑制率高达87.84％；在转染条件下，在5nM浓度的抑制率高达90.94％，在0.5nM浓度的抑制率高达84.78％。As can be seen from Table 15, the siRNA modifications provided by the present disclosure show excellent inhibitory effects on the ANGPTL3 gene. Under free uptake conditions, the inhibition rate at 100nM concentration is as high as 89.37%, and at 10nM concentration, the inhibition rate is as high as 87.84%; under transfection conditions, the inhibition rate at 5nM concentration is as high as 90.94%, and at 0.5nM concentration, the inhibition rate is as high as 90.94%. 84.78%.
实施例8：siRNA缀合物在人源化小鼠中对人ANGPTL3基因表达的抑制作用Example 8: Inhibitory effect of siRNA conjugates on human ANGPTL3 gene expression in humanized mice
6-8周龄的C57BL/6-hANGPTL3小鼠(由上海南方模式生物科技股份有限公司提供)进入饲养设施，适应性喂养7天后，以3mg/kg的单一剂量对小鼠分别皮下给药N-ER-FY013058M2L96、N-ER-FY013060M2L96、N-ER-FY013062M2L96L96和N-ER-FY013109M2L96(每组6只小鼠)。给药后第8日、第15日、第22日、第29日、第36日、第43日、第50日、第57日和第64日检测血清hANGPTL3蛋白表达量，由此得出siRNA缀合物对hANGPTL3蛋白表达的抑制率。C57BL/6-hANGPTL3 mice (provided by Shanghai Southern Model Biotechnology Co., Ltd.) aged 6-8 weeks entered the breeding facility. After adaptive feeding for 7 days, the mice were subcutaneously administered N at a single dose of 3 mg/kg. -ER-FY013058M2L96, N-ER-FY013060M2L96, N-ER-FY013062M2L96L96 and N-ER-FY013109M2L96 (6 mice per group). Serum hANGPTL3 protein expression was detected on the 8th, 15th, 22nd, 29th, 36th, 43rd, 50th, 57th and 64th days after administration, and siRNA was obtained. Inhibition rate of hANGPTL3 protein expression by the conjugate.
表16 siRNA缀合物对hANGPTL3蛋白抑制率
Table 16 Inhibition rate of hANGPTL3 protein by siRNA conjugates
从表16中可以看出，对于ANGPTL3基因的抑制率，本公开提供的siRNA缀合 物中，N-ER-FY013058M2L96、N-ER-FY013060M2L96和N-ER-FY013062M2L96在第8天至第43天的试验测定期间显示出抑制率高于88％的优异抑制效果，其中在第15天至第36天的抑制率均在90％以上，在第50天至第64天的试验测定期间，抑制率仍然保持在70％以上；对于N-ER-FY013109M2L96，在第8天至第43天中的抑制率均在75％以上，在第50天至第64天中的抑制率均高于50％。As can be seen from Table 16, for the inhibition rate of ANGPTL3 gene, the siRNA conjugated provided by the present disclosure Among the compounds, N-ER-FY013058M2L96, N-ER-FY013060M2L96 and N-ER-FY013062M2L96 showed excellent inhibitory effects with an inhibition rate higher than 88% during the test measurement period from day 8 to day 43, among which on day 15 The inhibition rate was above 90% to the 36th day, and the inhibition rate remained above 70% during the test measurement period from the 50th to the 64th day; for N-ER-FY013109M2L96, from the 8th to the 43rd day The inhibition rates were all above 75%, and the inhibition rates from days 50 to 64 were all above 50%.
实施例9 siRNA缀合物在CD-1小鼠血浆动力学研究Example 9 Kinetic study of siRNA conjugates in CD-1 mouse plasma
动物：CD-1小鼠，SPF级，雄性，30g左右，购买于斯贝福(北京)生物技术有限公司。Animal: CD-1 mouse, SPF grade, male, about 30g, purchased from Spefford (Beijing) Biotechnology Co., Ltd.
给药剂量和方式：3mg/kg(10mL/kg)，随机分组后单次皮下注射给药，6只小鼠每组。Dosage and method: 3mg/kg (10mL/kg), single subcutaneous injection after random grouping, 6 mice per group.
样品采集：采集给药后0.0833、0.25、0.5、1、2、4、8、24、36、48h全血样品，共10个点。每组前3只采集0.0833、0.5、2、8、36h，后3只采集0.25、1、4、24、48h，采集全血后离心分离血浆进行检测分析。Sample collection: Collect whole blood samples at 0.0833, 0.25, 0.5, 1, 2, 4, 8, 24, 36, and 48 hours after administration, a total of 10 points. The first three animals in each group were collected at 0.0833, 0.5, 2, 8, and 36 hours, and the last three animals were collected at 0.25, 1, 4, 24, and 48 hours. Whole blood was collected and plasma was centrifuged for detection and analysis.
样品检测与分析：采用LC-MS/MS方法检测各时间点血浆样品中原形药物的浓度，使用WinNonlin软件计算PK参数：Cmax、Tmax、AUC、MRT、t1/2。Sample detection and analysis: LC-MS/MS method was used to detect the concentration of prototype drug in plasma samples at each time point, and WinNonlin software was used to calculate PK parameters: C max , T max , AUC, MRT, t 1/2 .
从该实验中可以得出，本公开的siRNA缀合物在血浆中半衰期较短，清除较快。It can be concluded from this experiment that the siRNA conjugate of the present disclosure has a shorter half-life in plasma and is cleared faster.
实施例10 siRNA缀合物在CD-1小鼠组织分布试验Example 10 siRNA conjugate tissue distribution test in CD-1 mice
动物：CD-1小鼠，SPF级，雄性，30g左右，购买于斯贝福(北京)生物技术有限公司。Animal: CD-1 mouse, SPF grade, male, about 30g, purchased from Spefford (Beijing) Biotechnology Co., Ltd.
给药剂量和方式：3mg/kg(10mL/kg)，随机分组后单次皮下注射给药，每个时间点3只动物，共24只小鼠。Dosage and method: 3mg/kg (10mL/kg), single subcutaneous injection after randomization, 3 animals at each time point, 24 mice in total.
样品采集：Sample Collection:
给药后24h：采集血浆、肝、肾、脾；给药后72h：采集血浆、肝、肾、脾；24 hours after administration: collect plasma, liver, kidney, and spleen; 72 hours after administration: collect plasma, liver, kidney, and spleen;
给药后168h(1周)：采集血浆、肝、肾、脾、脑、心、肺、胃、小肠、肌肉、睾丸；给药后336h(2周)：采集血浆、肝、肾、脾；168h after administration (1 week): collect plasma, liver, kidney, spleen, brain, heart, lung, stomach, small intestine, muscle, and testes; 336h (2 weeks) after administration: collect plasma, liver, kidney, and spleen;
给药后672h(4周)：采集血浆、肝、肾、脾、脑、心、肺、胃、小肠、肌肉、睾丸；给药后1008h(6周)：采集血浆、肝、肾、脾；672h after administration (4 weeks): Collect plasma, liver, kidney, spleen, brain, heart, lung, stomach, small intestine, muscle, and testis; 1008h (6 weeks) after administration: Collect plasma, liver, kidney, and spleen;
给药后1344h(8周)：采集血浆、肝、肾、脾；1344h (8 weeks) after administration: plasma, liver, kidney, and spleen were collected;
给药后1680h(10周)：采集血浆、肝、肾、脾、脑、心、肺、胃、小肠、肌肉、睾丸。1680h (10 weeks) after administration: plasma, liver, kidney, spleen, brain, heart, lung, stomach, small intestine, muscle, and testis were collected.
样品检测与分析：采用LC-MS/MS方法检测各时间点血浆和组织样品中原形药物的浓度，采用梯形面积法计算血浆及组织中的AUC。Sample detection and analysis: The LC-MS/MS method was used to detect the concentration of the prototype drug in plasma and tissue samples at each time point, and the trapezoidal area method was used to calculate the AUC in plasma and tissue.
从该实验中可以得出，本公开的siRNA缀合物主要富集于肝脏，在组织中保留时间较长，具有很好的稳定性。It can be concluded from this experiment that the siRNA conjugate of the present disclosure is mainly enriched in the liver, has a long retention time in the tissue, and has good stability.
实施例11 siRNA缀合物单次皮下注射C57B/L小鼠给予MTD试验Example 11 Single subcutaneous injection of siRNA conjugate into C57B/L mice for MTD test
C57B/L小鼠，SPF级，雄性，25g左右，购买于斯贝福(北京)生物技术有限公司。动物根据适应期最后1天的体重，采用体重随机区组的方法，具体剂量设计和分组具体如下表17所示： C57B/L mice, SPF grade, male, about 25 g, were purchased from Spefford (Beijing) Biotechnology Co., Ltd. The animals were randomly grouped according to their body weight on the last day of the adaptation period. The specific dose design and grouping are shown in Table 17 below:
表17
Table 17
检测指标：Detection Indicator:
(1)临床观察：给药日连续观察4小时，恢复期每天至少进行一次临床观察(1) Clinical observation: Continuous observation for 4 hours on the dosing day, and at least one clinical observation per day during the recovery period
(2)体重：对所有存活动物每周进行2次体重称量。(2) Body weight: All surviving animals were weighed twice a week.
(3)免疫毒性：MTD剂量组动物于D1给药后1h±2min，4h±5min，8h±10min，24h±20min交替采血，每个时间点采集3只/性别/组动物，检测细胞因子(IFN-γ、TNF-α、IL-2/6/8)。(3) Immunotoxicity: Blood samples were collected from animals in the MTD dose group alternately at 1h±2min, 4h±5min, 8h±10min, and 24h±20min after D1 administration. 3 animals/sex/group were collected at each time point to detect cytokines ( IFN-γ, TNF-α, IL-2/6/8).
(4)毒代动力学：MTD剂量组动物于D1给药前、给药后30min±2min，1h±2min，4h±5min，8h±10min，24h±20min交替采血，每个时间点采集3只/性别/组动物，检测血药浓度。(4) Toxicokinetics: Blood samples were collected from animals in the MTD dose group alternately before D1 administration and at 30min±2min, 1h±2min, 4h±5min, 8h±10min, and 24h±20min after D1 administration. Three animals were collected at each time point. /Gender/ group of animals to detect blood drug concentration.
(5)血液生化学：主试验组动物于R28剖检，卫星组动物于R7、R14、R21、R28分批次剖检，检测血液生化学。(5) Blood biochemistry: The animals in the main experimental group were necropsied at R28, and the animals in the satellite group were necropsied in batches at R7, R14, R21, and R28 to detect blood biochemistry.
(6)组织分布：主试验组动物于R28剖检，卫星组动物于R7、R14、R21、R28分批次剖检，采集血、肝，检测组织药物浓度。(6) Tissue distribution: The animals in the main test group were necropsied at R28, and the animals in the satellite group were necropsied in batches at R7, R14, R21, and R28. Blood and liver were collected to detect tissue drug concentrations.
(7)组织病理学检查：主试验组动物于R28剖检，采集主要脏器(心、肝、脾、肺、肾、脑、肾上腺、胸腺、胃、子宫/睾丸、卵巢/附睾)以及发现异常的组织或脏器，取材并固定，进行组织病理学检查。(7) Histopathological examination: The animals in the main test group were necropsied at R28, and the main organs (heart, liver, spleen, lung, kidney, brain, adrenal gland, thymus, stomach, uterus/testis, ovary/epididymis) and findings were collected Abnormal tissues or organs are collected and fixed for histopathological examination.
从该实验中可以得出，本公开的siRNA缀合物毒性较低，具有优异的用药安全窗口。From this experiment, it can be concluded that the siRNA conjugate of the present disclosure has low toxicity and has an excellent drug safety window.
本公开的上述实施例仅是为清楚地说明本公开所作的举例，而并非是对本公开的实施方式的限定。对于所属领域的普通技术人员来说，在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。凡在本公开的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本公开权利要求的保护范围之内。 The above-described embodiments of the present disclosure are only examples to clearly illustrate the present disclosure, but are not intended to limit the implementation of the present disclosure. For those of ordinary skill in the art, other different forms of changes or modifications can be made based on the above description. An exhaustive list of all implementations is neither necessary nor possible. Any modifications, equivalent substitutions and improvements made within the spirit and principles of this disclosure shall be included in the protection scope of the claims of this disclosure.

Claims (45)

  1. 一种双链核糖核酸，所述双链核糖核酸包括正义链和反义链，所述正义链与所述反义链互补和/或基本上反向互补形成所述双链核糖核酸的双链区；A double-stranded ribonucleic acid, the double-stranded ribonucleic acid includes a sense strand and an antisense strand, the sense strand is complementary and/or substantially reverse complementary to the antisense strand to form the double strands of the double-stranded ribonucleic acid district;
    其中，所述正义链包含与靶标序列中至少15个连续核苷酸的差异不超过3个核苷酸的序列A，所述反义链包含与靶标序列中至少15个连续核苷酸的反向互补序列的差异不超过3个核苷酸的序列B；Wherein, the sense strand includes a sequence A that differs from at least 15 consecutive nucleotides in the target sequence by no more than 3 nucleotides, and the antisense strand includes a sequence A that differs from at least 15 consecutive nucleotides in the target sequence. Sequence B that differs from the complementary sequence by no more than 3 nucleotides;
    所述靶标序列选自如SEQ ID NO:1-7任一项所示的核苷酸序列。The target sequence is selected from the nucleotide sequence shown in any one of SEQ ID NO: 1-7.
  2. 根据权利要求1所述的双链核糖核酸，其中，所述靶标序列选自如SEQ ID NO:8-25任一项所示的核苷酸序列，所述正义链包含与SEQ ID NO:8-25任一项所示的核苷酸序列中至少15个连续核苷酸组成的序列相比差异不超过1个核苷酸的序列A，所述反义链包含与SEQ ID NO:8-25任一项所示的核苷酸序列中至少15个连续核苷酸组成的序列的反向互补序列相比差异不超过1个核苷酸的序列B；The double-stranded ribonucleic acid according to claim 1, wherein the target sequence is selected from the nucleotide sequence shown in any one of SEQ ID NO:8-25, and the sense strand includes the same sequence as SEQ ID NO:8-25. A sequence consisting of at least 15 consecutive nucleotides in any of the nucleotide sequences shown in 25, which differs by no more than 1 nucleotide from sequence A, and the antisense strand contains the same sequence as SEQ ID NO: 8-25 The reverse complement sequence of the sequence consisting of at least 15 consecutive nucleotides in the nucleotide sequence shown in any item is different from sequence B by no more than 1 nucleotide;
    可选地，所述差异的核苷酸位于所述序列A的3’末端；和/或，所述差异的核苷酸位于所述序列B的3’末端。Optionally, the different nucleotides are located at the 3' end of the sequence A; and/or the different nucleotides are located at the 3' end of the sequence B.
  3. 根据权利要求1或2所述的双链核糖核酸，其中，所述正义链由15-28个核苷酸组成，优选19-25个核苷酸，更优选19-23个核苷酸，更优选19、21或23个核苷酸。The double-stranded ribonucleic acid according to claim 1 or 2, wherein the sense strand consists of 15-28 nucleotides, preferably 19-25 nucleotides, more preferably 19-23 nucleotides, and more 19, 21 or 23 nucleotides are preferred.
  4. 根据权利要求3所述的双链核糖核酸，其中，所述正义链的核苷酸序列为与SEQ ID NO:8-25任一项所示的核苷酸序列中的15-28个连续核苷酸组成的序列相比差异不超过1个核苷酸的序列A，优选19-25个连续核苷酸，更优选19-23个连续核苷酸，更优选19、21或23个核苷酸。The double-stranded ribonucleic acid according to claim 3, wherein the nucleotide sequence of the sense strand is 15-28 consecutive nuclei in the nucleotide sequence shown in any one of SEQ ID NO:8-25 Compared with sequence A that differs by no more than 1 nucleotide, the sequence composed of nucleotides is preferably 19-25 consecutive nucleotides, more preferably 19-23 consecutive nucleotides, and more preferably 19, 21 or 23 nucleosides. acid.
  5. 根据权利要求1-4任一项所述的双链核糖核酸，其中，所述反义链由15-28个核苷酸组成，优选19-25个核苷酸，更优选19-23个核苷酸，更优选19、21或23个核苷酸。The double-stranded ribonucleic acid according to any one of claims 1-4, wherein the antisense strand consists of 15-28 nucleotides, preferably 19-25 nucleotides, more preferably 19-23 nucleotides nucleotides, more preferably 19, 21 or 23 nucleotides.
  6. 根据权利要求5所述的双链核糖核酸，其中，所述反义链的核苷酸序列是与SEQ ID NO:8-25任一项所示的核苷酸序列中的15-28个连续核苷酸组成的序列的反向互补序列相比差异不超过1个核苷酸的序列B，优选19-25个连续核苷酸，更优选19-23个连续核苷酸，更优选19、21或23个核苷酸。The double-stranded ribonucleic acid according to claim 5, wherein the nucleotide sequence of the antisense strand is 15-28 consecutive with the nucleotide sequence shown in any one of SEQ ID NO:8-25 The reverse complementary sequence of the sequence composed of nucleotides is preferably 19-25 consecutive nucleotides, more preferably 19-23 consecutive nucleotides, and more preferably 19, 21 or 23 nucleotides.
  7. 根据权利要求1-6任一项所述的双链核糖核酸，其中，所述双链区的长度为15-25个核苷酸，优选19-23个核苷酸，更优选19-21个核苷酸，更优选19、21或23个核苷酸。The double-stranded ribonucleic acid according to any one of claims 1-6, wherein the length of the double-stranded region is 15-25 nucleotides, preferably 19-23 nucleotides, more preferably 19-21 nucleotides nucleotides, more preferably 19, 21 or 23 nucleotides.
  8. 根据权利要求1-7任一项所述的双链核糖核酸，其中，The double-stranded ribonucleic acid according to any one of claims 1-7, wherein,
    所述正义链与所述反义链互补形成所述双链区，且所述正义链的3’末端具有1-2个延伸出所述双链区的突出的核苷酸，所述反义链的3’末端形成平末端；或者，The sense strand and the antisense strand are complementary to form the double-stranded region, and the 3' end of the sense strand has 1-2 protruding nucleotides extending out of the double-stranded region, and the antisense strand is complementary to the double-stranded region. The 3' end of the chain forms a blunt end; or,
    所述正义链与所述反义链互补形成所述双链区，且所述反义链的3’末端具有1-2个延伸出所述双链区的突出的核苷酸，所述正义链的3’末端形成平末端；或者，The sense strand and the antisense strand are complementary to form the double-stranded region, and the 3' end of the antisense strand has 1-2 protruding nucleotides extending out of the double-stranded region, and the sense strand The 3' end of the chain forms a blunt end; or,
    所述正义链与所述反义链互补形成所述双链区，且所述正义链与所述反义链的3’末端均具有1-2个延伸出所述双链区的突出的核苷酸；或者，The sense strand and the antisense strand are complementary to form the double-stranded region, and the 3' ends of the sense strand and the antisense strand each have 1-2 protruding cores extending out of the double-stranded region. glycoside; or,
    所述正义链与所述反义链互补形成所述双链区，且所述正义链与所述反义链的3’末端均形成平末端。The sense strand and the antisense strand are complementary to form the double-stranded region, and the 3' ends of the sense strand and the antisense strand both form blunt ends.
  9. 根据权利要求1-8任一项所述的双链核糖核酸，其中，所述正义链与所述反义链选自如下组合：The double-stranded ribonucleic acid according to any one of claims 1-8, wherein the sense strand and the antisense strand are selected from the following combinations:
    82)所述正义链包含如SEQ ID NO:107所示的核苷酸序列，所述反义链包含如SEQ ID NO:202所示的核苷酸序列；82) The sense strand includes the nucleotide sequence shown in SEQ ID NO:107, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:202;
    1)所述正义链包含如SEQ ID NO:26所示的核苷酸序列，所述反义链包含如SEQ ID NO:121所示的核苷酸序列；1) The sense strand includes the nucleotide sequence shown in SEQ ID NO:26, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:121;
    2)所述正义链包含如SEQ ID NO:27所示的核苷酸序列，所述反义链包含如SEQ ID NO:122所示的核苷酸序列；2) The sense strand includes the nucleotide sequence shown in SEQ ID NO:27, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:122;
    3)所述正义链包含如SEQ ID NO:28所示的核苷酸序列，所述反义链包含如SEQ ID  NO:123所示的核苷酸序列；3) The sense strand includes the nucleotide sequence shown in SEQ ID NO:28, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:28 The nucleotide sequence shown in NO:123;
    4)所述正义链包含如SEQ ID NO:29所示的核苷酸序列，所述反义链包含如SEQ ID NO:124所示的核苷酸序列；4) The sense strand includes the nucleotide sequence shown in SEQ ID NO:29, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:124;
    5)所述正义链包含如SEQ ID NO:30所示的核苷酸序列，所述反义链包含如SEQ ID NO:125所示的核苷酸序列；5) The sense strand includes the nucleotide sequence shown in SEQ ID NO:30, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:125;
    6)所述正义链包含如SEQ ID NO:31所示的核苷酸序列，所述反义链包含如SEQ ID NO:126所示的核苷酸序列；6) The sense strand includes the nucleotide sequence shown in SEQ ID NO:31, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:126;
    7)所述正义链包含如SEQ ID NO:32所示的核苷酸序列，所述反义链包含如SEQ ID NO:127所示的核苷酸序列；7) The sense strand includes the nucleotide sequence shown in SEQ ID NO:32, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:127;
    8)所述正义链包含如SEQ ID NO:33所示的核苷酸序列，所述反义链包含如SEQ ID NO:128所示的核苷酸序列；8) The sense strand includes the nucleotide sequence shown in SEQ ID NO:33, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:128;
    9)所述正义链包含如SEQ ID NO:34所示的核苷酸序列，所述反义链包含如SEQ ID NO:129所示的核苷酸序列；9) The sense strand includes the nucleotide sequence shown in SEQ ID NO:34, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:129;
    10)所述正义链包含如SEQ ID NO:35所示的核苷酸序列，所述反义链包含如SEQ ID NO:130所示的核苷酸序列；10) The sense strand includes the nucleotide sequence shown in SEQ ID NO:35, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:130;
    11)所述正义链包含如SEQ ID NO:36所示的核苷酸序列，所述反义链包含如SEQ ID NO:131所示的核苷酸序列；11) The sense strand includes the nucleotide sequence shown in SEQ ID NO:36, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:131;
    12)所述正义链包含如SEQ ID NO:37所示的核苷酸序列，所述反义链包含如SEQ ID NO:132所示的核苷酸序列；12) The sense strand includes the nucleotide sequence shown in SEQ ID NO:37, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:132;
    13)所述正义链包含如SEQ ID NO:38所示的核苷酸序列，所述反义链包含如SEQ ID NO:133所示的核苷酸序列；13) The sense strand includes the nucleotide sequence shown in SEQ ID NO:38, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:133;
    14)所述正义链包含如SEQ ID NO:39所示的核苷酸序列，所述反义链包含如SEQ ID NO:134所示的核苷酸序列；14) The sense strand includes the nucleotide sequence shown in SEQ ID NO:39, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:134;
    15)所述正义链包含如SEQ ID NO:40所示的核苷酸序列，所述反义链包含如SEQ ID NO:135所示的核苷酸序列；15) The sense strand includes the nucleotide sequence shown in SEQ ID NO:40, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:135;
    16)所述正义链包含如SEQ ID NO:41所示的核苷酸序列，所述反义链包含如SEQ ID NO:136所示的核苷酸序列；16) The sense strand includes the nucleotide sequence shown in SEQ ID NO:41, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:136;
    17)所述正义链包含如SEQ ID NO:42所示的核苷酸序列，所述反义链包含如SEQ ID NO:137所示的核苷酸序列；17) The sense strand includes the nucleotide sequence shown in SEQ ID NO:42, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:137;
    18)所述正义链包含如SEQ ID NO:43所示的核苷酸序列，所述反义链包含如SEQ ID NO:138所示的核苷酸序列；18) The sense strand includes the nucleotide sequence shown in SEQ ID NO:43, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:138;
    19)所述正义链包含如SEQ ID NO:44所示的核苷酸序列，所述反义链包含如SEQ ID NO:139所示的核苷酸序列；19) The sense strand includes the nucleotide sequence shown in SEQ ID NO:44, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:139;
    20)所述正义链包含如SEQ ID NO:45所示的核苷酸序列，所述反义链包含如SEQ ID NO:140所示的核苷酸序列；20) The sense strand includes the nucleotide sequence shown in SEQ ID NO:45, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:140;
    21)所述正义链包含如SEQ ID NO:46所示的核苷酸序列，所述反义链包含如SEQ ID NO:141所示的核苷酸序列；21) The sense strand includes the nucleotide sequence shown in SEQ ID NO:46, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:141;
    22)所述正义链包含如SEQ ID NO:47所示的核苷酸序列，所述反义链包含如SEQ ID NO:142所示的核苷酸序列；22) The sense strand includes the nucleotide sequence shown in SEQ ID NO:47, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:142;
    23)所述正义链包含如SEQ ID NO:48所示的核苷酸序列，所述反义链包含如SEQ ID NO:143所示的核苷酸序列；23) The sense strand includes the nucleotide sequence shown in SEQ ID NO:48, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:143;
    24)所述正义链包含如SEQ ID NO:49所示的核苷酸序列，所述反义链包含如SEQ ID NO:144所示的核苷酸序列；24) The sense strand includes the nucleotide sequence shown in SEQ ID NO:49, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:144;
    25)所述正义链包含如SEQ ID NO:50所示的核苷酸序列，所述反义链包含如SEQ ID NO:145所示的核苷酸序列； 25) The sense strand includes the nucleotide sequence shown in SEQ ID NO:50, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:145;
    26)所述正义链包含如SEQ ID NO:51所示的核苷酸序列，所述反义链包含如SEQ ID NO:146所示的核苷酸序列；26) The sense strand includes the nucleotide sequence shown in SEQ ID NO:51, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:146;
    27)所述正义链包含如SEQ ID NO:52所示的核苷酸序列，所述反义链包含如SEQ ID NO:147所示的核苷酸序列；27) The sense strand includes the nucleotide sequence shown in SEQ ID NO:52, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:147;
    28)所述正义链包含如SEQ ID NO:53所示的核苷酸序列，所述反义链包含如SEQ ID NO:148所示的核苷酸序列；28) The sense strand includes the nucleotide sequence shown in SEQ ID NO:53, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:148;
    29)所述正义链包含如SEQ ID NO:54所示的核苷酸序列，所述反义链包含如SEQ ID NO:149所示的核苷酸序列；29) The sense strand includes the nucleotide sequence shown in SEQ ID NO:54, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:149;
    30)所述正义链包含如SEQ ID NO:55所示的核苷酸序列，所述反义链包含如SEQ ID NO:150所示的核苷酸序列；30) The sense strand includes the nucleotide sequence shown in SEQ ID NO:55, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:150;
    31)所述正义链包含如SEQ ID NO:56所示的核苷酸序列，所述反义链包含如SEQ ID NO:151所示的核苷酸序列；31) The sense strand includes the nucleotide sequence shown in SEQ ID NO:56, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:151;
    32)所述正义链包含如SEQ ID NO:57所示的核苷酸序列，所述反义链包含如SEQ ID NO:152所示的核苷酸序列；32) The sense strand includes the nucleotide sequence shown in SEQ ID NO:57, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:152;
    33)所述正义链包含如SEQ ID NO:58所示的核苷酸序列，所述反义链包含如SEQ ID NO:153所示的核苷酸序列；33) The sense strand includes the nucleotide sequence shown in SEQ ID NO:58, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:153;
    34)所述正义链包含如SEQ ID NO:59所示的核苷酸序列，所述反义链包含如SEQ ID NO:154所示的核苷酸序列；34) The sense strand includes the nucleotide sequence shown in SEQ ID NO:59, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:154;
    35)所述正义链包含如SEQ ID NO:60所示的核苷酸序列，所述反义链包含如SEQ ID NO:155所示的核苷酸序列；35) The sense strand includes the nucleotide sequence shown in SEQ ID NO:60, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:155;
    36)所述正义链包含如SEQ ID NO:61所示的核苷酸序列，所述反义链包含如SEQ ID NO:156所示的核苷酸序列；36) The sense strand includes the nucleotide sequence shown in SEQ ID NO:61, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:156;
    37)所述正义链包含如SEQ ID NO:62所示的核苷酸序列，所述反义链包含如SEQ ID NO:157所示的核苷酸序列；37) The sense strand includes the nucleotide sequence shown in SEQ ID NO:62, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:157;
    38)所述正义链包含如SEQ ID NO:63所示的核苷酸序列，所述反义链包含如SEQ ID NO:158所示的核苷酸序列；38) The sense strand includes the nucleotide sequence shown in SEQ ID NO: 63, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 158;
    39)所述正义链包含如SEQ ID NO:64所示的核苷酸序列，所述反义链包含如SEQ ID NO:159所示的核苷酸序列；39) The sense strand includes the nucleotide sequence shown in SEQ ID NO:64, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:159;
    40)所述正义链包含如SEQ ID NO:65所示的核苷酸序列，所述反义链包含如SEQ ID NO:160所示的核苷酸序列；40) The sense strand includes the nucleotide sequence shown in SEQ ID NO:65, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:160;
    41)所述正义链包含如SEQ ID NO:66所示的核苷酸序列，所述反义链包含如SEQ ID NO:161所示的核苷酸序列；41) The sense strand includes the nucleotide sequence shown in SEQ ID NO:66, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:161;
    42)所述正义链包含如SEQ ID NO:67所示的核苷酸序列，所述反义链包含如SEQ ID NO:162所示的核苷酸序列；42) The sense strand includes the nucleotide sequence shown in SEQ ID NO:67, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:162;
    43)所述正义链包含如SEQ ID NO:68所示的核苷酸序列，所述反义链包含如SEQ ID NO:163所示的核苷酸序列；43) The sense strand includes the nucleotide sequence shown in SEQ ID NO:68, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:163;
    44)所述正义链包含如SEQ ID NO:69所示的核苷酸序列，所述反义链包含如SEQ ID NO:164所示的核苷酸序列；44) The sense strand includes the nucleotide sequence shown in SEQ ID NO:69, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:164;
    45)所述正义链包含如SEQ ID NO:70所示的核苷酸序列，所述反义链包含如SEQ ID NO:165所示的核苷酸序列；45) The sense strand includes the nucleotide sequence shown in SEQ ID NO:70, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:165;
    46)所述正义链包含如SEQ ID NO:71所示的核苷酸序列，所述反义链包含如SEQ ID NO:166所示的核苷酸序列；46) The sense strand includes the nucleotide sequence shown in SEQ ID NO:71, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:166;
    47)所述正义链包含如SEQ ID NO:72所示的核苷酸序列，所述反义链包含如SEQ ID NO:167所示的核苷酸序列；47) The sense strand includes the nucleotide sequence shown in SEQ ID NO:72, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:167;
    48)所述正义链包含如SEQ ID NO:73所示的核苷酸序列，所述反义链包含如SEQ ID  NO:168所示的核苷酸序列；48) The sense strand includes the nucleotide sequence shown in SEQ ID NO:73, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:73 The nucleotide sequence shown in NO:168;
    49)所述正义链包含如SEQ ID NO:74所示的核苷酸序列，所述反义链包含如SEQ ID NO:169所示的核苷酸序列；49) The sense strand includes the nucleotide sequence shown in SEQ ID NO:74, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:169;
    50)所述正义链包含如SEQ ID NO:75所示的核苷酸序列，所述反义链包含如SEQ ID NO:170所示的核苷酸序列；50) The sense strand includes the nucleotide sequence shown in SEQ ID NO:75, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:170;
    51)所述正义链包含如SEQ ID NO:76所示的核苷酸序列，所述反义链包含如SEQ ID NO:171所示的核苷酸序列；51) The sense strand includes the nucleotide sequence shown in SEQ ID NO:76, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:171;
    52)所述正义链包含如SEQ ID NO:77所示的核苷酸序列，所述反义链包含如SEQ ID NO:172所示的核苷酸序列；52) The sense strand includes the nucleotide sequence shown in SEQ ID NO:77, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:172;
    53)所述正义链包含如SEQ ID NO:78所示的核苷酸序列，所述反义链包含如SEQ ID NO:173所示的核苷酸序列；53) The sense strand includes the nucleotide sequence shown in SEQ ID NO:78, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:173;
    54)所述正义链包含如SEQ ID NO:79所示的核苷酸序列，所述反义链包含如SEQ ID NO:174所示的核苷酸序列；54) The sense strand includes the nucleotide sequence shown in SEQ ID NO:79, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:174;
    55)所述正义链包含如SEQ ID NO:80所示的核苷酸序列，所述反义链包含如SEQ ID NO:175所示的核苷酸序列；55) The sense strand includes the nucleotide sequence shown in SEQ ID NO:80, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:175;
    56)所述正义链包含如SEQ ID NO:81所示的核苷酸序列，所述反义链包含如SEQ ID NO:176所示的核苷酸序列；56) The sense strand includes the nucleotide sequence shown in SEQ ID NO:81, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:176;
    57)所述正义链包含如SEQ ID NO:82所示的核苷酸序列，所述反义链包含如SEQ ID NO:177所示的核苷酸序列；57) The sense strand includes the nucleotide sequence shown in SEQ ID NO:82, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:177;
    58)所述正义链包含如SEQ ID NO:83所示的核苷酸序列，所述反义链包含如SEQ ID NO:178所示的核苷酸序列；58) The sense strand includes the nucleotide sequence shown in SEQ ID NO:83, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:178;
    59)所述正义链包含如SEQ ID NO:84所示的核苷酸序列，所述反义链包含如SEQ ID NO:179所示的核苷酸序列；59) The sense strand includes the nucleotide sequence shown in SEQ ID NO:84, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:179;
    60)所述正义链包含如SEQ ID NO:85所示的核苷酸序列，所述反义链包含如SEQ ID NO:180所示的核苷酸序列；60) The sense strand includes the nucleotide sequence shown in SEQ ID NO:85, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:180;
    61)所述正义链包含如SEQ ID NO:86所示的核苷酸序列，所述反义链包含如SEQ ID NO:181所示的核苷酸序列；61) The sense strand includes the nucleotide sequence shown in SEQ ID NO:86, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:181;
    62)所述正义链包含如SEQ ID NO:87所示的核苷酸序列，所述反义链包含如SEQ ID NO:182所示的核苷酸序列；62) The sense strand includes the nucleotide sequence shown in SEQ ID NO:87, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:182;
    63)所述正义链包含如SEQ ID NO:88所示的核苷酸序列，所述反义链包含如SEQ ID NO:183所示的核苷酸序列；63) The sense strand includes the nucleotide sequence shown in SEQ ID NO:88, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:183;
    64)所述正义链包含如SEQ ID NO:89所示的核苷酸序列，所述反义链包含如SEQ ID NO:184所示的核苷酸序列；64) The sense strand includes the nucleotide sequence shown in SEQ ID NO:89, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:184;
    65)所述正义链包含如SEQ ID NO:90所示的核苷酸序列，所述反义链包含如SEQ ID NO:185所示的核苷酸序列；65) The sense strand includes the nucleotide sequence shown in SEQ ID NO:90, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:185;
    66)所述正义链包含如SEQ ID NO:91所示的核苷酸序列，所述反义链包含如SEQ ID NO:186所示的核苷酸序列；66) The sense strand includes the nucleotide sequence shown in SEQ ID NO:91, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:186;
    67)所述正义链包含如SEQ ID NO:92所示的核苷酸序列，所述反义链包含如SEQ ID NO:187所示的核苷酸序列；67) The sense strand includes the nucleotide sequence shown in SEQ ID NO:92, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:187;
    68)所述正义链包含如SEQ ID NO:93所示的核苷酸序列，所述反义链包含如SEQ ID NO:188所示的核苷酸序列；68) The sense strand includes the nucleotide sequence shown in SEQ ID NO:93, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:188;
    69)所述正义链包含如SEQ ID NO:94所示的核苷酸序列，所述反义链包含如SEQ ID NO:189所示的核苷酸序列；69) The sense strand includes the nucleotide sequence shown in SEQ ID NO:94, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:189;
    70)所述正义链包含如SEQ ID NO:95所示的核苷酸序列，所述反义链包含如SEQ ID NO:190所示的核苷酸序列； 70) The sense strand includes the nucleotide sequence shown in SEQ ID NO:95, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:190;
    71)所述正义链包含如SEQ ID NO:96所示的核苷酸序列，所述反义链包含如SEQ ID NO:191所示的核苷酸序列；71) The sense strand includes the nucleotide sequence shown in SEQ ID NO:96, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:191;
    72)所述正义链包含如SEQ ID NO:97所示的核苷酸序列，所述反义链包含如SEQ ID NO:192所示的核苷酸序列；72) The sense strand includes the nucleotide sequence shown in SEQ ID NO:97, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:192;
    73)所述正义链包含如SEQ ID NO:98所示的核苷酸序列，所述反义链包含如SEQ ID NO:193所示的核苷酸序列；73) The sense strand includes the nucleotide sequence shown in SEQ ID NO:98, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:193;
    74)所述正义链包含如SEQ ID NO:99所示的核苷酸序列，所述反义链包含如SEQ ID NO:194所示的核苷酸序列；74) The sense strand includes the nucleotide sequence shown in SEQ ID NO:99, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:194;
    75)所述正义链包含如SEQ ID NO:100所示的核苷酸序列，所述反义链包含如SEQ ID NO:195所示的核苷酸序列；75) The sense strand includes the nucleotide sequence shown in SEQ ID NO:100, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:195;
    76)所述正义链包含如SEQ ID NO:101所示的核苷酸序列，所述反义链包含如SEQ ID NO:196所示的核苷酸序列；76) The sense strand includes the nucleotide sequence shown in SEQ ID NO:101, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:196;
    77)所述正义链包含如SEQ ID NO:102所示的核苷酸序列，所述反义链包含如SEQ ID NO:197所示的核苷酸序列；77) The sense strand includes the nucleotide sequence shown in SEQ ID NO:102, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:197;
    78)所述正义链包含如SEQ ID NO:103所示的核苷酸序列，所述反义链包含如SEQ ID NO:198所示的核苷酸序列；78) The sense strand includes the nucleotide sequence shown in SEQ ID NO:103, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:198;
    79)所述正义链包含如SEQ ID NO:104所示的核苷酸序列，所述反义链包含如SEQ ID NO:199所示的核苷酸序列；79) The sense strand includes the nucleotide sequence shown in SEQ ID NO:104, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:199;
    80)所述正义链包含如SEQ ID NO:105所示的核苷酸序列，所述反义链包含如SEQ ID NO:200所示的核苷酸序列；80) The sense strand includes the nucleotide sequence shown in SEQ ID NO:105, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:200;
    81)所述正义链包含如SEQ ID NO:106所示的核苷酸序列，所述反义链包含如SEQ ID NO:201所示的核苷酸序列；81) The sense strand includes the nucleotide sequence shown in SEQ ID NO:106, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:201;
    83)所述正义链包含如SEQ ID NO:108所示的核苷酸序列，所述反义链包含如SEQ ID NO:203所示的核苷酸序列；83) The sense strand includes the nucleotide sequence shown in SEQ ID NO:108, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:203;
    84)所述正义链包含如SEQ ID NO:109所示的核苷酸序列，所述反义链包含如SEQ ID NO:204所示的核苷酸序列；84) The sense strand includes the nucleotide sequence shown in SEQ ID NO:109, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:204;
    85)所述正义链包含如SEQ ID NO:110所示的核苷酸序列，所述反义链包含如SEQ ID NO:205所示的核苷酸序列；85) The sense strand includes the nucleotide sequence shown in SEQ ID NO:110, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:205;
    86)所述正义链包含如SEQ ID NO:111所示的核苷酸序列，所述反义链包含如SEQ ID NO:206所示的核苷酸序列；86) The sense strand includes the nucleotide sequence shown in SEQ ID NO:111, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:206;
    87)所述正义链包含如SEQ ID NO:112所示的核苷酸序列，所述反义链包含如SEQ ID NO:207所示的核苷酸序列；87) The sense strand includes the nucleotide sequence shown in SEQ ID NO:112, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:207;
    88)所述正义链包含如SEQ ID NO:113所示的核苷酸序列，所述反义链包含如SEQ ID NO:208所示的核苷酸序列；88) The sense strand includes the nucleotide sequence shown in SEQ ID NO:113, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:208;
    89)所述正义链包含如SEQ ID NO:114所示的核苷酸序列，所述反义链包含如SEQ ID NO:209所示的核苷酸序列；89) The sense strand includes the nucleotide sequence shown in SEQ ID NO:114, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:209;
    90)所述正义链包含如SEQ ID NO:115所示的核苷酸序列，所述反义链包含如SEQ ID NO:210所示的核苷酸序列；90) The sense strand includes the nucleotide sequence shown in SEQ ID NO:115, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:210;
    91)所述正义链包含如SEQ ID NO:116所示的核苷酸序列，所述反义链包含如SEQ ID NO:211所示的核苷酸序列；91) The sense strand includes the nucleotide sequence shown in SEQ ID NO:116, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:211;
    92)所述正义链包含如SEQ ID NO:117所示的核苷酸序列，所述反义链包含如SEQ ID NO:212所示的核苷酸序列；92) The sense strand includes the nucleotide sequence shown in SEQ ID NO:117, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:212;
    93)所述正义链包含如SEQ ID NO:118所示的核苷酸序列，所述反义链包含如SEQ ID NO:213所示的核苷酸序列；93) The sense strand includes the nucleotide sequence shown in SEQ ID NO:118, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:213;
    94)所述正义链包含如SEQ ID NO:119所示的核苷酸序列，所述反义链包含如SEQ ID  NO:214所示的核苷酸序列；94) The sense strand includes the nucleotide sequence shown in SEQ ID NO:119, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:119 The nucleotide sequence shown in NO:214;
    95)所述正义链包含如SEQ ID NO:120所示的核苷酸序列，所述反义链包含如SEQ ID NO:215所示的核苷酸序列；95) The sense strand includes the nucleotide sequence shown in SEQ ID NO:120, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:215;
    96)所述正义链包含如SEQ ID NO:258所示的核苷酸序列，所述反义链包含如SEQ ID NO:268所示的核苷酸序列；96) The sense strand includes the nucleotide sequence shown in SEQ ID NO:258, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:268;
    97)所述正义链包含如SEQ ID NO:259所示的核苷酸序列，所述反义链包含如SEQ ID NO:269所示的核苷酸序列；97) The sense strand includes the nucleotide sequence shown in SEQ ID NO:259, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:269;
    98)所述正义链包含如SEQ ID NO:260所示的核苷酸序列，所述反义链包含如SEQ ID NO:270所示的核苷酸序列；98) The sense strand includes the nucleotide sequence shown in SEQ ID NO:260, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:270;
    99)所述正义链包含如SEQ ID NO:261所示的核苷酸序列，所述反义链包含如SEQ ID NO:271所示的核苷酸序列；99) The sense strand includes the nucleotide sequence shown in SEQ ID NO:261, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:271;
    100)所述正义链包含如SEQ ID NO:262所示的核苷酸序列，所述反义链包含如SEQ ID NO:272所示的核苷酸序列；100) The sense strand includes the nucleotide sequence shown in SEQ ID NO:262, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:272;
    101)所述正义链包含如SEQ ID NO:263所示的核苷酸序列，所述反义链包含如SEQ ID NO:273所示的核苷酸序列；101) The sense strand includes the nucleotide sequence shown in SEQ ID NO:263, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:273;
    102)所述正义链包含如SEQ ID NO:264所示的核苷酸序列，所述反义链包含如SEQ ID NO:274所示的核苷酸序列；102) The sense strand includes the nucleotide sequence shown in SEQ ID NO:264, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:274;
    103)所述正义链包含如SEQ ID NO:265所示的核苷酸序列，所述反义链包含如SEQ ID NO:275所示的核苷酸序列；103) The sense strand includes the nucleotide sequence shown in SEQ ID NO:265, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:275;
    104)所述正义链包含如SEQ ID NO:266所示的核苷酸序列，所述反义链包含如SEQ ID NO:276所示的核苷酸序列；104) The sense strand includes the nucleotide sequence shown in SEQ ID NO:266, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:276;
    105)所述正义链包含如SEQ ID NO:267所示的核苷酸序列，所述反义链包含如SEQ ID NO:277所示的核苷酸序列；105) The sense strand includes the nucleotide sequence shown in SEQ ID NO:267, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:277;
    137)所述正义链包含如SEQ ID NO:298所示的核苷酸序列，所述反义链包含如SEQ ID NO:321所示的核苷酸序列；137) The sense strand includes the nucleotide sequence shown in SEQ ID NO:298, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:321;
    138)所述正义链包含如SEQ ID NO:299所示的核苷酸序列，所述反义链包含如SEQ ID NO:322所示的核苷酸序列；138) The sense strand includes the nucleotide sequence shown in SEQ ID NO:299, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:322;
    139)所述正义链包含如SEQ ID NO:300所示的核苷酸序列，所述反义链包含如SEQ ID NO:323所示的核苷酸序列；139) The sense strand includes the nucleotide sequence shown in SEQ ID NO:300, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:323;
    140)所述正义链包含如SEQ ID NO:301所示的核苷酸序列，所述反义链包含如SEQ ID NO:324所示的核苷酸序列；140) The sense strand includes the nucleotide sequence shown in SEQ ID NO:301, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:324;
    141)所述正义链包含如SEQ ID NO:302所示的核苷酸序列，所述反义链包含如SEQ ID NO:325所示的核苷酸序列；141) The sense strand includes the nucleotide sequence shown in SEQ ID NO:302, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:325;
    142)所述正义链包含如SEQ ID NO:303所示的核苷酸序列，所述反义链包含如SEQ ID NO:326所示的核苷酸序列；142) The sense strand includes the nucleotide sequence shown in SEQ ID NO:303, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:326;
    143)所述正义链包含如SEQ ID NO:304所示的核苷酸序列，所述反义链包含如SEQ ID NO:327所示的核苷酸序列；143) The sense strand includes the nucleotide sequence shown in SEQ ID NO:304, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:327;
    144)所述正义链包含如SEQ ID NO:305所示的核苷酸序列，所述反义链包含如SEQ ID NO:328所示的核苷酸序列；144) The sense strand includes the nucleotide sequence shown in SEQ ID NO:305, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:328;
    145)所述正义链包含如SEQ ID NO:306所示的核苷酸序列，所述反义链包含如SEQ ID NO:329所示的核苷酸序列；145) The sense strand includes the nucleotide sequence shown in SEQ ID NO:306, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:329;
    146)所述正义链包含如SEQ ID NO:307所示的核苷酸序列，所述反义链包含如SEQ ID NO:330所示的核苷酸序列；146) The sense strand includes the nucleotide sequence shown in SEQ ID NO:307, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:330;
    147)所述正义链包含如SEQ ID NO:308所示的核苷酸序列，所述反义链包含如SEQ ID NO:331所示的核苷酸序列； 147) The sense strand includes the nucleotide sequence shown in SEQ ID NO:308, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:331;
    148)所述正义链包含如SEQ ID NO:309所示的核苷酸序列，所述反义链包含如SEQ ID NO:332所示的核苷酸序列；148) The sense strand includes the nucleotide sequence shown in SEQ ID NO:309, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:332;
    149)所述正义链包含如SEQ ID NO:310所示的核苷酸序列，所述反义链包含如SEQ ID NO:333所示的核苷酸序列；149) The sense strand includes the nucleotide sequence shown in SEQ ID NO:310, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:333;
    150)所述正义链包含如SEQ ID NO:311所示的核苷酸序列，所述反义链包含如SEQ ID NO:334所示的核苷酸序列；150) The sense strand includes the nucleotide sequence shown in SEQ ID NO:311, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:334;
    151)所述正义链包含如SEQ ID NO:312所示的核苷酸序列，所述反义链包含如SEQ ID NO:335所示的核苷酸序列；151) The sense strand includes the nucleotide sequence shown in SEQ ID NO:312, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:335;
    152)所述正义链包含如SEQ ID NO:313所示的核苷酸序列，所述反义链包含如SEQ ID NO:336所示的核苷酸序列；152) The sense strand includes the nucleotide sequence shown in SEQ ID NO:313, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:336;
    153)所述正义链包含如SEQ ID NO:314所示的核苷酸序列，所述反义链包含如SEQ ID NO:337所示的核苷酸序列；153) The sense strand includes the nucleotide sequence shown in SEQ ID NO:314, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:337;
    154)所述正义链包含如SEQ ID NO:315所示的核苷酸序列，所述反义链包含如SEQ ID NO:338所示的核苷酸序列；154) The sense strand includes the nucleotide sequence shown in SEQ ID NO:315, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:338;
    155)所述正义链包含如SEQ ID NO:316所示的核苷酸序列，所述反义链包含如SEQ ID NO:339所示的核苷酸序列；155) The sense strand includes the nucleotide sequence shown in SEQ ID NO:316, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:339;
    156)所述正义链包含如SEQ ID NO:317所示的核苷酸序列，所述反义链包含如SEQ ID NO:340所示的核苷酸序列；156) The sense strand includes the nucleotide sequence shown in SEQ ID NO:317, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:340;
    157)所述正义链包含如SEQ ID NO:318所示的核苷酸序列，所述反义链包含如SEQ ID NO:341所示的核苷酸序列；157) The sense strand includes the nucleotide sequence shown in SEQ ID NO:318, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:341;
    158)所述正义链包含如SEQ ID NO:319所示的核苷酸序列，所述反义链包含如SEQ ID NO:342所示的核苷酸序列；158) The sense strand includes the nucleotide sequence shown in SEQ ID NO:319, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:342;
    159)所述正义链包含如SEQ ID NO:320所示的核苷酸序列，所述反义链包含如SEQ ID NO:343所示的核苷酸序列；159) The sense strand includes the nucleotide sequence shown in SEQ ID NO:320, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:343;
    201)所述正义链包含如SEQ ID NO:385所示的核苷酸序列，所述反义链包含如SEQ ID NO:387所示的核苷酸序列；201) The sense strand includes the nucleotide sequence shown in SEQ ID NO:385, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:387;
    202)所述正义链包含如SEQ ID NO:386所示的核苷酸序列，所述反义链包含如SEQ ID NO:388所示的核苷酸序列。202) The sense strand includes the nucleotide sequence shown in SEQ ID NO:386, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:388.
  10. 根据权利要求1-9任一项所述的双链核糖核酸，其中，所述正义链中每个核苷酸彼此独立地为修饰的核苷酸或未修饰的核苷酸，和/或，所述反义链中每个核苷酸彼此独立地为修饰的核苷酸或未修饰的核苷酸。The double-stranded ribonucleic acid according to any one of claims 1 to 9, wherein each nucleotide in the sense strand is independently a modified nucleotide or an unmodified nucleotide, and/or, Each nucleotide in the antisense strand is independently a modified nucleotide or an unmodified nucleotide.
  11. 根据权利要求1-10任一项所述的双链核糖核酸，其中，所述正义链中任意相连的两个核苷酸由磷酸二酯键或硫代磷酸二酯键连接，和/或，所述反义链中任意相连的两个核苷酸由磷酸二酯键或硫代磷酸二酯键连接。The double-stranded ribonucleic acid according to any one of claims 1 to 10, wherein any two connected nucleotides in the sense strand are connected by a phosphodiester bond or a phosphorothioate diester bond, and/or, Any two connected nucleotides in the antisense strand are connected by a phosphodiester bond or a phosphorothioate diester bond.
  12. 根据权利要求1-11任一项所述的双链核糖核酸，其中，所述正义链的5’末端核苷酸连接5’磷酸基团或5’磷酸衍生基团，和/或，所述反义链的5’末端核苷酸连接5’磷酸基团或5’磷酸衍生基团。The double-stranded ribonucleic acid according to any one of claims 1-11, wherein the 5' terminal nucleotide of the sense strand is connected to a 5' phosphate group or a 5' phosphate derivative group, and/or, the The 5' terminal nucleotide of the antisense strand is linked to a 5' phosphate group or a 5' phosphate derivative group.
  13. 根据权利要求1-12任一项所述的双链核糖核酸，其中，所述双链核糖核酸为siRNA。The double-stranded ribonucleic acid according to any one of claims 1-12, wherein the double-stranded ribonucleic acid is siRNA.
  14. 根据权利要求1-13任一项所述的双链核糖核酸，其中，所述双链核糖核酸为用于抑制ANGPTL3基因表达的siRNA。The double-stranded ribonucleic acid according to any one of claims 1 to 13, wherein the double-stranded ribonucleic acid is siRNA for inhibiting ANGPTL3 gene expression.
  15. 一种双链核糖核酸修饰物，其为如权利要求1-14任一项所述的双链核糖核酸的修饰物，所述双链核糖核酸修饰物包含如下至少一种的化学修饰：A double-stranded ribonucleic acid modification, which is a double-stranded ribonucleic acid modification as described in any one of claims 1-14, wherein the double-stranded ribonucleic acid modification includes at least one of the following chemical modifications:
    (1)正义链中至少一个核苷酸的修饰，(1) Modification of at least one nucleotide in the sense strand,
    (2)正义链中至少一个位置处的磷酸二酯键的修饰， (2) Modification of the phosphodiester bond at at least one position in the sense strand,
    (3)反义链中至少一个核苷酸的修饰，(3) Modification of at least one nucleotide in the antisense strand,
    (4)反义链中至少一个位置处的磷酸二酯键的修饰；(4) Modification of the phosphodiester bond at at least one position in the antisense strand;
    任选地，所述双链核糖核酸的正义链中序列A的3’末端连接由1-2个核苷酸组成的序列D，优选由1-2个胸腺嘧啶脱氧核糖核苷酸组成的序列D；和/或，所述双链核糖核酸的反义链中序列B的3’末端连接由1-2个核苷酸组成的序列E，优选由1-2个胸腺嘧啶脱氧核糖核苷酸组成的序列E；和/或，所述双链核糖核酸的正义链中序列A的3’末端排除1-2个核苷酸后形成序列A’；Optionally, the 3' end of sequence A in the sense strand of the double-stranded ribonucleic acid is connected to sequence D consisting of 1-2 nucleotides, preferably a sequence consisting of 1-2 thymidine deoxyribonucleotides D; and/or, the 3' end of sequence B in the antisense strand of the double-stranded ribonucleic acid is connected to sequence E consisting of 1-2 nucleotides, preferably 1-2 thymidine deoxyribonucleotides Sequence E consisting of; and/or sequence A' is formed after excluding 1-2 nucleotides from the 3' end of sequence A in the sense strand of the double-stranded ribonucleic acid;
    可选地，所述双链核糖核酸修饰物的正义链和反义链选自如下的序列组合：Alternatively, the sense strand and antisense strand of the double-stranded ribonucleic acid modification are selected from the following sequence combinations:
    所述正义链的核苷酸序列为序列A所示的序列，所述反义链的核苷酸序列为序列B所示的序列；The nucleotide sequence of the sense strand is the sequence shown in sequence A, and the nucleotide sequence of the antisense strand is the sequence shown in sequence B;
    或者，所述正义链的核苷酸序列为序列A所示的序列，所述反义链的核苷酸序列为序列B连接序列E所示的序列；Alternatively, the nucleotide sequence of the sense strand is the sequence shown in sequence A, and the nucleotide sequence of the antisense strand is the sequence shown in sequence B connected to sequence E;
    或者，所述正义链的核苷酸序列为序列A连接序列D所示的序列，所述反义链的核苷酸序列为序列B所示的序列；Alternatively, the nucleotide sequence of the sense strand is the sequence shown in sequence A connected to sequence D, and the nucleotide sequence of the antisense strand is the sequence shown in sequence B;
    或者，所述正义链的核苷酸序列为序列A连接序列D所示的序列，所述反义链的核苷酸序列为序列B连接序列E所示的序列；Alternatively, the nucleotide sequence of the sense strand is the sequence shown in sequence A connected to sequence D, and the nucleotide sequence of the antisense strand is the sequence shown in sequence B connected to sequence E;
    或者，所述正义链的核苷酸序列为序列A’所示的序列，所述反义链的核苷酸序列为序列B所示的序列；Alternatively, the nucleotide sequence of the sense strand is the sequence shown in sequence A', and the nucleotide sequence of the antisense strand is the sequence shown in sequence B;
    或者，所述正义链的核苷酸序列为序列A’所示的序列，所述反义链的核苷酸序列为序列B连接序列E所示的序列。Alternatively, the nucleotide sequence of the sense strand is the sequence represented by sequence A', and the nucleotide sequence of the antisense strand is the sequence represented by sequence B connected to sequence E.
  16. 根据权利要求15所述的双链核糖核酸修饰物，其中，所述核苷酸的修饰选自2’-氟代修饰，2’-烷氧基修饰，2’-取代的烷氧基修饰，2’-烷基修饰，2’-取代的烷基修饰，2’-脱氧修饰，核苷酸衍生物修饰或其中任意两种以上的组合。The double-stranded ribonucleic acid modification according to claim 15, wherein the modification of the nucleotide is selected from the group consisting of 2'-fluoro modification, 2'-alkoxy modification, and 2'-substituted alkoxy modification, 2'-alkyl modification, 2'-substituted alkyl modification, 2'-deoxy modification, nucleotide derivative modification or a combination of any two or more thereof.
  17. 根据权利要求15或16所述的双链核糖核酸修饰物，其中，所述核苷酸的修饰选自2’-F修饰、2’-O-CH3修饰、2’-O-CH2-CH2-O-CH3修饰、2’-O-CH2-CH＝CH2修饰、2’-CH2-CH2-CH＝CH2修饰、2’-脱氧修饰，核苷酸衍生物修饰或其中任意两种以上的组合。The double-stranded ribonucleic acid modification according to claim 15 or 16, wherein the modification of the nucleotide is selected from the group consisting of 2'-F modification, 2'-O-CH 3 modification, and 2'-O-CH 2 - CH 2 -O-CH 3 modification, 2'-O-CH 2 -CH＝CH 2 modification, 2'-CH 2 -CH 2 -CH＝CH 2 modification, 2'-deoxy modification, nucleotide derivative modification Or a combination of any two or more of them.
  18. 根据权利要求16或17所述的双链核糖核酸修饰物，其中，所述核苷酸衍生物修饰中的核苷酸衍生物选自异核苷酸、LNA、ENA、cET、UNA或GNA。The double-stranded ribonucleic acid modification according to claim 16 or 17, wherein the nucleotide derivative in the nucleotide derivative modification is selected from isonucleotides, LNA, ENA, cET, UNA or GNA.
  19. 根据权利要求15-18任一项所述的双链核糖核酸修饰物，其中，沿5’末端向3’末端方向，所述正义链中第7位、第9位、第10位和第11位的核糖核苷酸为2’-F修饰的核糖核苷酸，所述正义链中其余位置的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸。The double-stranded ribonucleic acid modification according to any one of claims 15 to 18, wherein, along the 5' end to the 3' end direction, the 7th, 9th, 10th and 11th positions in the sense strand The ribonucleotides at the position are 2'-F modified ribonucleotides, and the ribonucleotides at the remaining positions in the sense strand are 2'-O- CH3 modified ribonucleotides.
  20. 根据权利要求15-19任一项所述的双链核糖核酸修饰物，其中，沿5’末端向3’末端方向，所述正义链包含位于如下所示位置处的硫代磷酸二酯键：The double-stranded ribonucleic acid modification according to any one of claims 15-19, wherein, along the 5' end to the 3' end direction, the sense strand includes a phosphorothioate diester bond located at the position shown below:
    所述正义链5’末端起始的第1个核苷酸与第2个核苷酸之间；Between the first nucleotide and the second nucleotide starting from the 5' end of the sense strand;
    所述正义链5’末端起始的第2个核苷酸与第3个核苷酸之间；Between the 2nd and 3rd nucleotide starting from the 5' end of the sense strand;
    所述正义链3’末端起始的第1个核苷酸与第2个核苷酸之间；Between the first nucleotide and the second nucleotide starting from the 3' end of the sense strand;
    所述正义链3’末端起始的第2个核苷酸与第3个核苷酸之间；Between the second and third nucleotides starting from the 3’ end of the sense strand;
    或者，or,
    所述正义链包含位于如下所示位置处的硫代磷酸二酯键：The sense strand contains phosphorothioate diester bonds at the positions shown below:
    所述正义链5’末端起始的第1个核苷酸与第2个核苷酸之间；Between the first nucleotide and the second nucleotide starting from the 5' end of the sense strand;
    所述正义链5’末端起始的第2个核苷酸与第3个核苷酸之间。Between the 2nd and 3rd nucleotide starting from the 5' end of the sense strand.
  21. 根据权利要求15-20任一项所述的双链核糖核酸修饰物，其中，沿5’末端向3’末端方向，所述反义链中任意奇数位置处的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸，所述反义链中任意偶数位置处的核糖核苷酸为2’-F修饰的核糖核苷酸；The double-stranded ribonucleic acid modification according to any one of claims 15 to 20, wherein, along the direction from the 5' end to the 3' end, the ribonucleotide at any odd-numbered position in the antisense strand is 2'- O- CH3 modified ribonucleotide, the ribonucleotide at any even-numbered position in the antisense strand is a 2'-F modified ribonucleotide;
    或者，沿5’末端向3’末端方向，所述反义链中第2位、第6位、第14位和第16位的核糖 核苷酸为2’-F修饰的核糖核苷酸，所述反义链中其余位置的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸；Alternatively, along the 5' end to the 3' end, the ribose sugars at positions 2, 6, 14 and 16 in the antisense strand The nucleotides are 2'-F modified ribonucleotides, and the remaining ribonucleotides in the antisense strand are 2'-O- CH3 modified ribonucleotides;
    或者，沿5’末端向3’末端方向，所述反义链中第2位、第6位、第8位、第9位、第14位和第16位的核糖核苷酸为2’-F修饰的核糖核苷酸，所述反义链中其余位置的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸；Alternatively, along the direction from the 5' end to the 3' end, the ribonucleotides at positions 2, 6, 8, 9, 14 and 16 in the antisense strand are 2'- F-modified ribonucleotides, the remaining ribonucleotides in the antisense strand are 2'-O-CH 3 modified ribonucleotides;
    或者，沿5’末端向3’末端方向，所述反义链中第2位、第14位和第16位的核糖核苷酸为2’-F修饰的核糖核苷酸，所述反义链中第6位的核糖核苷酸为核苷酸衍生物GNA修饰的核糖核苷酸，所述反义链中其余位置的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸；Alternatively, along the direction from the 5' end to the 3' end, the ribonucleotides at positions 2, 14 and 16 in the antisense strand are 2'-F modified ribonucleotides, and the antisense strand The ribonucleotide at position 6 in the chain is a ribonucleotide modified by the nucleotide derivative GNA, and the ribonucleotides at the remaining positions in the antisense chain are ribonucleotides modified by 2'-O-CH 3 acid;
    或者，沿5’末端向3’末端方向，所述反义链中第2位、第6位、第14位和第16位的核糖核苷酸为2’-F修饰的核糖核苷酸，所述反义链中第7位的核糖核苷酸为核苷酸衍生物GNA修饰的核糖核苷酸，所述反义链中其余位置的核糖核苷酸为2’-O-CH3修饰的核糖核苷酸。Alternatively, along the direction from the 5' end to the 3' end, the ribonucleotides at positions 2, 6, 14 and 16 in the antisense strand are 2'-F modified ribonucleotides, The ribonucleotide at position 7 in the antisense strand is a ribonucleotide modified by the nucleotide derivative GNA, and the ribonucleotides at the remaining positions in the antisense strand are modified with 2'-O-CH 3 of ribonucleotides.
  22. 根据权利要求15-21任一项所述的双链核糖核酸修饰物，其中，沿5’末端向3’末端方向，所述反义链的5’末端的核苷酸连接5’磷酸基团或5’磷酸衍生基团。The double-stranded ribonucleic acid modification according to any one of claims 15 to 21, wherein the nucleotide at the 5' end of the antisense strand is connected to a 5' phosphate group along the 5' end to the 3' end. or 5' phosphate derivatized group.
  23. 根据权利要求15-22任一项所述的双链核糖核酸修饰物，其中，所述反义链包含位于如下所示位置处的硫代磷酸二酯键：The double-stranded ribonucleic acid modification according to any one of claims 15-22, wherein the antisense strand includes a phosphorothioate diester bond located at the position shown below:
    所述反义链5’末端起始的第1个核苷酸与第2个核苷酸之间；Between the first nucleotide and the second nucleotide starting from the 5' end of the antisense strand;
    所述反义链5’末端起始的第2个核苷酸与第3个核苷酸之间；Between the 2nd and 3rd nucleotide starting from the 5' end of the antisense strand;
    所述反义链3’末端起始的第1个核苷酸与第2个核苷酸之间；Between the first nucleotide and the second nucleotide starting from the 3' end of the antisense strand;
    所述反义链3’末端起始的第2个核苷酸与第3个核苷酸之间。Between the 2nd and 3rd nucleotide starting from the 3' end of the antisense strand.
  24. 根据权利要求15-23任一项所述的双链核糖核酸修饰物，其中，所述双链核糖核酸修饰物的正义链具有如(a1)-(a5)任一项所示的结构：The double-stranded ribonucleic acid modification according to any one of claims 15-23, wherein the sense strand of the double-stranded ribonucleic acid modification has a structure shown in any one of (a 1 ) to (a 5 ) :
    (a1)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-(s)-T-(s)-T-3’，(a 1 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3',
    (a2)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(a 2 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
    (a3)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(a 3 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3',
    (a4)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-3’，(a 4 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -3',
    (a5)5’-mN1-(s)-mN2-(s)-mN3-mN4-mN5-mN6-N7f-mN8-N9f-N10f-N11f-mN12-mN13-mN14-mN15-mN16-mN17-mN18-mN19-mN20-mN21-3’；(a 5 )5'-mN 1 -(s)-mN 2 -(s)-mN 3 -mN 4 -mN 5 -mN 6 -N 7 f-mN 8 -N 9 fN 10 fN 11 f-mN 12 -mN 13 -mN 14 -mN 15 -mN 16 -mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -3';
    其中，N1-N23彼此独立地选自碱基为A、U、C或G的核糖核苷酸，Wherein, N 1 -N 23 are independently selected from ribonucleotides whose bases are A, U, C or G,
    大写字母T表示碱基为胸腺嘧啶的脱氧核糖核苷酸，The capital letter T stands for deoxyribonucleotide whose base is thymine.
    小写字母m表示该字母m右侧相邻的一个核糖核苷酸为2’-O-CH3修饰的核糖核苷酸，The lowercase letter m indicates that the ribonucleotide adjacent to the right side of the letter m is a 2'-O-CH 3 modified ribonucleotide.
    小写字母f表示该字母f左侧相邻的一个核糖核苷酸为2’-F修饰的核糖核苷酸，The lowercase letter f indicates that the ribonucleotide adjacent to the left side of the letter f is a 2’-F modified ribonucleotide.
    -(s)-表示前后相邻的两个核苷酸以硫代磷酸二酯键连接。-(s)- indicates that two adjacent nucleotides are connected by a phosphorothioate diester bond.
  25. 根据权利要求15-24任一项所述的双链核糖核酸修饰物，其中，所述双链核糖核酸修饰物的反义链具有如(b1)-(b15)任一项所示的结构：The double-stranded ribonucleic acid modification according to any one of claims 15 to 24, wherein the antisense strand of the double-stranded ribonucleic acid modification has a structure as shown in any one of (b 1 ) to (b 15 ) structure:
    (b1)5’-P1mN1-(s)-N2f-(s)-mN3-N4f-mN5-N6f-mN7-N8f-mN9-N10f-mN11-N12f-mN13-N14f-mN15-N16f-mN17-N18f-mN19-(s)-T-(s)-T-3’，(b 1 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -N 4 f-mN 5 -N 6 f-mN 7 -N 8 f-mN 9 -N 10 f- mN 11 -N 12 f-mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -N 18 f-mN 19 -(s)-T-(s)-T-3',
    (b2)5’-P1mN1-(s)-N2f-(s)-mN3-N4f-mN5-N6f-mN7-N8f-mN9-N10f-mN11-N12f-mN13-N14f-mN15-N16f-mN17-N18f-mN19-(s)-N20f-(s)-mN21-3’，(b 2 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -N 4 f-mN 5 -N 6 f-mN 7 -N 8 f-mN 9 -N 10 f- mN 11 -N 12 f-mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -N 18 f-mN 19 -(s)-N 20 f-(s)-mN 21 -3',
    (b3)5’-P1mN1-(s)-N2f-(s)-mN3-N4f-mN5-N6f-mN7-N8f-mN9-N10f-mN11-N12f-mN13-N14f-mN15-N16f-mN17-N18f-mN19-N20f-mN21-(s)-N22f-(s)-mN23-3’,(b 3 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -N 4 f-mN 5 -N 6 f-mN 7 -N 8 f-mN 9 -N 10 f- mN 11 -N 12 f-mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -N 18 f-mN 19 -N 20 f-mN 21 -(s)-N 22 f-(s)- mN 23 -3',
    (b4)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N1 4f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 4 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -mN 8 -mN 9 -mN 10 -mN 11 - mN 12 -mN 13 -N 1 4 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3',
    (b5)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N1 4f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 5 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -mN 8 -mN 9 -mN 10 -mN 11 - mN 12 -mN 13 -N 1 4 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
    (b6)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N1 4f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(b 6 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -mN 8 -mN 9 -mN 10 -mN 11 - mN 12 -mN 13 -N 1 4 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3',
    (b7)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-N8f-N9f-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 7 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -N 8 fN 9 f-mN 10 -mN 11 - mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3'，
    (b8)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-N8f-N9f-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 8 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -N 8 fN 9 f-mN 10 -mN 11 - mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
    (b9)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-mN7-N8f-N9f-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(b 9 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-mN 7 -N 8 fN 9 f-mN 10 -mN 11 - mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3',
    (b10)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-[GNA]N6-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 10 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -[GNA]N 6 -mN 7 -mN 8 -mN 9 -mN 10 -mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3',
    (b11)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-[GNA]N6-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 11 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -[GNA]N 6 -mN 7 -mN 8 -mN 9 -mN 10 -mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
    (b12)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-[GNA]N6-mN7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(b 12 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -[GNA]N 6 -mN 7 -mN 8 -mN 9 -mN 10 -mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3' ,
    (b13)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-[GNA]N7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-T-(s)-T-3’，(b 13 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-[GNA]N 7 -mN 8 -mN 9 -mN 10 - mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-T-(s)-T-3',
    (b14)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-[GNA]N7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-(s)-mN20-(s)-mN21-3’，(b 14 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-[GNA]N 7 -mN 8 -mN 9 -mN 10 - mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -(s)-mN 20 -(s)-mN 21 -3',
    (b15)5’-P1mN1-(s)-N2f-(s)-mN3-mN4-mN5-N6f-[GNA]N7-mN8-mN9-mN10-mN11-mN12-mN13-N14f-mN15-N16f-mN17-mN18-mN19-mN20-mN21-(s)-mN22-(s)-mN23-3’，(b 15 )5'-P1mN 1 -(s)-N 2 f-(s)-mN 3 -mN 4 -mN 5 -N 6 f-[GNA]N 7 -mN 8 -mN 9 -mN 10 - mN 11 -mN 12 -mN 13 -N 14 f-mN 15 -N 16 f-mN 17 -mN 18 -mN 19 -mN 20 -mN 21 -(s)-mN 22 -(s)-mN 23 -3 ',
    其中，N1-N23彼此独立地选自碱基为A、U、C或G的核糖核苷酸，Wherein, N 1 -N 23 are independently selected from ribonucleotides whose bases are A, U, C or G,
    大写字母T表示碱基为胸腺嘧啶的脱氧核糖核苷酸，The capital letter T stands for deoxyribonucleotide whose base is thymine.
    小写字母m表示该字母m右侧相邻的一个核糖核苷酸为2’-O-CH3修饰的核糖核苷酸，The lowercase letter m indicates that the ribonucleotide adjacent to the right side of the letter m is a 2'-O-CH 3 modified ribonucleotide.
    小写字母f表示该字母f左侧相邻的一个核糖核苷酸为2’-F修饰的核糖核苷酸，The lowercase letter f indicates that the ribonucleotide adjacent to the left side of the letter f is a 2’-F modified ribonucleotide.
    P1表示该字母右侧相邻的一个核苷酸为5’-磷酸核苷酸，P1 indicates that the nucleotide adjacent to the right side of the letter is a 5’-phosphate nucleotide,
    -(s)-表示前后相邻的两个核苷酸以硫代磷酸二酯键连接，-(s)- means that two adjacent nucleotides are connected by a phosphorothioate diester bond,
    [GNA]表示其右侧相邻的一个核糖核苷酸为存在GNA修饰的核糖核苷酸。[GNA] indicates that the ribonucleotide adjacent to the right side is a ribonucleotide modified by GNA.
  26. 根据权利要求15-25任一项所述的双链核糖核酸修饰物，其中，所述双链核糖核酸修饰物为siRNA修饰物。The double-stranded ribonucleic acid modified product according to any one of claims 15 to 25, wherein the double-stranded ribonucleic acid modified product is an siRNA modified product.
  27. 根据权利要求15-26任一项所述的双链核糖核酸修饰物，其中，所述双链核糖核酸修饰物为用于抑制ANGPTL3基因表达的siRNA修饰物。The double-stranded ribonucleic acid modification according to any one of claims 15 to 26, wherein the double-stranded ribonucleic acid modification is an siRNA modification for inhibiting ANGPTL3 gene expression.
  28. 根据权利要求15-27任一项所述的双链核糖核酸修饰物，其中，所述正义链与所述反义链选自如下组合：The double-stranded ribonucleic acid modification according to any one of claims 15-27, wherein the sense strand and the antisense strand are selected from the following combinations:
    106)所述正义链包含如SEQ ID NO:216所示的核苷酸序列，所述反义链包含如SEQ ID NO:237所示的核苷酸序列；106) The sense strand includes the nucleotide sequence shown in SEQ ID NO:216, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:237;
    107)所述正义链包含如SEQ ID NO:217所示的核苷酸序列，所述反义链包含如SEQ ID NO:238所示的核苷酸序列；107) The sense strand includes the nucleotide sequence shown in SEQ ID NO:217, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:238;
    108)所述正义链包含如SEQ ID NO:218所示的核苷酸序列，所述反义链包含如SEQ ID NO:239所示的核苷酸序列；108) The sense strand includes the nucleotide sequence shown in SEQ ID NO:218, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:239;
    109)所述正义链包含如SEQ ID NO:219所示的核苷酸序列，所述反义链包含如SEQ ID NO:240所示的核苷酸序列；109) The sense strand includes the nucleotide sequence shown in SEQ ID NO:219, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:240;
    110)所述正义链包含如SEQ ID NO:220所示的核苷酸序列，所述反义链包含如SEQ ID  NO:241所示的核苷酸序列；110) The sense strand includes the nucleotide sequence shown in SEQ ID NO:220, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:220 The nucleotide sequence shown in NO:241;
    111)所述正义链包含如SEQ ID NO:221所示的核苷酸序列，所述反义链包含如SEQ ID NO:242所示的核苷酸序列；111) The sense strand includes the nucleotide sequence shown in SEQ ID NO:221, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:242;
    112)所述正义链包含如SEQ ID NO:222所示的核苷酸序列，所述反义链包含如SEQ ID NO:243所示的核苷酸序列；112) The sense strand includes the nucleotide sequence shown in SEQ ID NO:222, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:243;
    113)所述正义链包含如SEQ ID NO:223所示的核苷酸序列，所述反义链包含如SEQ ID NO:244所示的核苷酸序列；113) The sense strand includes the nucleotide sequence shown in SEQ ID NO:223, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:244;
    114)所述正义链包含如SEQ ID NO:224所示的核苷酸序列，所述反义链包含如SEQ ID NO:245所示的核苷酸序列；114) The sense strand includes the nucleotide sequence shown in SEQ ID NO:224, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:245;
    115)所述正义链包含如SEQ ID NO:225所示的核苷酸序列，所述反义链包含如SEQ ID NO:246所示的核苷酸序列；115) The sense strand includes the nucleotide sequence shown in SEQ ID NO:225, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:246;
    116)所述正义链包含如SEQ ID NO:226所示的核苷酸序列，所述反义链包含如SEQ ID NO:247所示的核苷酸序列；116) The sense strand includes the nucleotide sequence shown in SEQ ID NO:226, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:247;
    117)所述正义链包含如SEQ ID NO:227所示的核苷酸序列，所述反义链包含如SEQ ID NO:248所示的核苷酸序列；117) The sense strand includes the nucleotide sequence shown in SEQ ID NO:227, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:248;
    118)所述正义链包含如SEQ ID NO:228所示的核苷酸序列，所述反义链包含如SEQ ID NO:249所示的核苷酸序列；118) The sense strand includes the nucleotide sequence shown in SEQ ID NO:228, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:249;
    119)所述正义链包含如SEQ ID NO:229所示的核苷酸序列，所述反义链包含如SEQ ID NO:250所示的核苷酸序列；119) The sense strand includes the nucleotide sequence shown in SEQ ID NO:229, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:250;
    120)所述正义链包含如SEQ ID NO:230所示的核苷酸序列，所述反义链包含如SEQ ID NO:251所示的核苷酸序列；120) The sense strand includes the nucleotide sequence shown in SEQ ID NO:230, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:251;
    121)所述正义链包含如SEQ ID NO:231所示的核苷酸序列，所述反义链包含如SEQ ID NO:252所示的核苷酸序列；121) The sense strand includes the nucleotide sequence shown in SEQ ID NO:231, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:252;
    122)所述正义链包含如SEQ ID NO:232所示的核苷酸序列，所述反义链包含如SEQ ID NO:253所示的核苷酸序列；122) The sense strand includes the nucleotide sequence shown in SEQ ID NO:232, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:253;
    123)所述正义链包含如SEQ ID NO:233所示的核苷酸序列，所述反义链包含如SEQ ID NO:254所示的核苷酸序列；123) The sense strand includes the nucleotide sequence shown in SEQ ID NO:233, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:254;
    124)所述正义链包含如SEQ ID NO:234所示的核苷酸序列，所述反义链包含如SEQ ID NO:241所示的核苷酸序列；124) The sense strand includes the nucleotide sequence shown in SEQ ID NO:234, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:241;
    125)所述正义链包含如SEQ ID NO:235所示的核苷酸序列，所述反义链包含如SEQ ID NO:247所示的核苷酸序列；125) The sense strand includes the nucleotide sequence shown in SEQ ID NO:235, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:247;
    126)所述正义链包含如SEQ ID NO:236所示的核苷酸序列，所述反义链包含如SEQ ID NO:253所示的核苷酸序列；126) The sense strand includes the nucleotide sequence shown in SEQ ID NO:236, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:253;
    127)所述正义链包含如SEQ ID NO:278所示的核苷酸序列，所述反义链包含如SEQ ID NO:288所示的核苷酸序列；127) The sense strand includes the nucleotide sequence shown in SEQ ID NO:278, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:288;
    128)所述正义链包含如SEQ ID NO:279所示的核苷酸序列，所述反义链包含如SEQ ID NO:289所示的核苷酸序列；128) The sense strand includes the nucleotide sequence shown in SEQ ID NO:279, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:289;
    129)所述正义链包含如SEQ ID NO:280所示的核苷酸序列，所述反义链包含如SEQ ID NO:290所示的核苷酸序列；129) The sense strand includes the nucleotide sequence shown in SEQ ID NO:280, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:290;
    130)所述正义链包含如SEQ ID NO:281所示的核苷酸序列，所述反义链包含如SEQ ID NO:291所示的核苷酸序列；130) The sense strand includes the nucleotide sequence shown in SEQ ID NO:281, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:291;
    131)所述正义链包含如SEQ ID NO:282所示的核苷酸序列，所述反义链包含如SEQ ID NO:292所示的核苷酸序列；131) The sense strand includes the nucleotide sequence shown in SEQ ID NO:282, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:292;
    132)所述正义链包含如SEQ ID NO:283所示的核苷酸序列，所述反义链包含如SEQ ID NO:293所示的核苷酸序列； 132) The sense strand includes the nucleotide sequence shown in SEQ ID NO:283, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:293;
    133)所述正义链包含如SEQ ID NO:284所示的核苷酸序列，所述反义链包含如SEQ ID NO:294所示的核苷酸序列；133) The sense strand includes the nucleotide sequence shown in SEQ ID NO:284, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:294;
    134)所述正义链包含如SEQ ID NO:285所示的核苷酸序列，所述反义链包含如SEQ ID NO:295所示的核苷酸序列；134) The sense strand includes the nucleotide sequence shown in SEQ ID NO:285, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:295;
    135)所述正义链包含如SEQ ID NO:286所示的核苷酸序列，所述反义链包含如SEQ ID NO:296所示的核苷酸序列；135) The sense strand includes the nucleotide sequence shown in SEQ ID NO:286, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:296;
    136)所述正义链包含如SEQ ID NO:287所示的核苷酸序列，所述反义链包含如SEQ ID NO:297所示的核苷酸序列；136) The sense strand includes the nucleotide sequence shown in SEQ ID NO:287, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:297;
    160)所述正义链包含如SEQ ID NO:344所示的核苷酸序列，所述反义链包含如SEQ ID NO:354所示的核苷酸序列；160) The sense strand includes the nucleotide sequence shown in SEQ ID NO:344, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:354;
    161)所述正义链包含如SEQ ID NO:345所示的核苷酸序列，所述反义链包含如SEQ ID NO:355所示的核苷酸序列；161) The sense strand includes the nucleotide sequence shown in SEQ ID NO:345, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:355;
    162)所述正义链包含如SEQ ID NO:346所示的核苷酸序列，所述反义链包含如SEQ ID NO:356所示的核苷酸序列；162) The sense strand includes the nucleotide sequence shown in SEQ ID NO:346, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:356;
    163)所述正义链包含如SEQ ID NO:347所示的核苷酸序列，所述反义链包含如SEQ ID NO:357所示的核苷酸序列；163) The sense strand includes the nucleotide sequence shown in SEQ ID NO:347, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:357;
    164)所述正义链包含如SEQ ID NO:348所示的核苷酸序列，所述反义链包含如SEQ ID NO:358所示的核苷酸序列；164) The sense strand includes the nucleotide sequence shown in SEQ ID NO:348, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:358;
    165)所述正义链包含如SEQ ID NO:349所示的核苷酸序列，所述反义链包含如SEQ ID NO:359所示的核苷酸序列；165) The sense strand includes the nucleotide sequence shown in SEQ ID NO:349, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:359;
    166)所述正义链包含如SEQ ID NO:350所示的核苷酸序列，所述反义链包含如SEQ ID NO:360所示的核苷酸序列；166) The sense strand includes the nucleotide sequence shown in SEQ ID NO:350, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:360;
    167)所述正义链包含如SEQ ID NO:351所示的核苷酸序列，所述反义链包含如SEQ ID NO:361所示的核苷酸序列；167) The sense strand includes the nucleotide sequence shown in SEQ ID NO:351, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:361;
    168)所述正义链包含如SEQ ID NO:231所示的核苷酸序列，所述反义链包含如SEQ ID NO:362所示的核苷酸序列；168) The sense strand includes the nucleotide sequence shown in SEQ ID NO:231, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:362;
    169)所述正义链包含如SEQ ID NO:352所示的核苷酸序列，所述反义链包含如SEQ ID NO:362所示的核苷酸序列；169) The sense strand includes the nucleotide sequence shown in SEQ ID NO:352, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:362;
    170)所述正义链包含如SEQ ID NO:231所示的核苷酸序列，所述反义链包含如SEQ ID NO:363所示的核苷酸序列；170) The sense strand includes the nucleotide sequence shown in SEQ ID NO:231, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:363;
    171)所述正义链包含如SEQ ID NO:231所示的核苷酸序列，所述反义链包含如SEQ ID NO:364所示的核苷酸序列；171) The sense strand includes the nucleotide sequence shown in SEQ ID NO:231, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:364;
    172)所述正义链包含如SEQ ID NO:231所示的核苷酸序列，所述反义链包含如SEQ ID NO:365所示的核苷酸序列；172) The sense strand includes the nucleotide sequence shown in SEQ ID NO:231, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:365;
    173)所述正义链包含如SEQ ID NO:232所示的核苷酸序列，所述反义链包含如SEQ ID NO:366所示的核苷酸序列；173) The sense strand includes the nucleotide sequence shown in SEQ ID NO:232, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:366;
    174)所述正义链包含如SEQ ID NO:236所示的核苷酸序列，所述反义链包含如SEQ ID NO:366所示的核苷酸序列；174) The sense strand includes the nucleotide sequence shown in SEQ ID NO:236, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:366;
    175)所述正义链包含如SEQ ID NO:232所示的核苷酸序列，所述反义链包含如SEQ ID NO:367所示的核苷酸序列；175) The sense strand includes the nucleotide sequence shown in SEQ ID NO:232, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:367;
    176)所述正义链包含如SEQ ID NO:232所示的核苷酸序列，所述反义链包含如SEQ ID NO:368所示的核苷酸序列；176) The sense strand includes the nucleotide sequence shown in SEQ ID NO:232, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:368;
    177)所述正义链包含如SEQ ID NO:232所示的核苷酸序列，所述反义链包含如SEQ ID NO:369所示的核苷酸序列；177) The sense strand includes the nucleotide sequence shown in SEQ ID NO:232, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:369;
    178)所述正义链包含如SEQ ID NO:233所示的核苷酸序列，所述反义链包含如SEQ ID  NO:370所示的核苷酸序列；178) The sense strand comprises the nucleotide sequence shown in SEQ ID NO:233, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:233 The nucleotide sequence shown in NO:370;
    179)所述正义链包含如SEQ ID NO:353所示的核苷酸序列，所述反义链包含如SEQ ID NO:370所示的核苷酸序列；179) The sense strand includes the nucleotide sequence shown in SEQ ID NO:353, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:370;
    180)所述正义链包含如SEQ ID NO:233所示的核苷酸序列，所述反义链包含如SEQ ID NO:371所示的核苷酸序列；180) The sense strand includes the nucleotide sequence shown in SEQ ID NO:233, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:371;
    181)所述正义链包含如SEQ ID NO:233所示的核苷酸序列，所述反义链包含如SEQ ID NO:372所示的核苷酸序列；181) The sense strand includes the nucleotide sequence shown in SEQ ID NO:233, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:372;
    182)所述正义链包含如SEQ ID NO:233所示的核苷酸序列，所述反义链包含如SEQ ID NO:373所示的核苷酸序列；182) The sense strand includes the nucleotide sequence shown in SEQ ID NO:233, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:373;
    183)所述正义链包含如SEQ ID NO:389所示的核苷酸序列，所述反义链包含如SEQ ID NO:381所示的核苷酸序列；183) The sense strand includes the nucleotide sequence shown in SEQ ID NO:389, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:381;
    184)所述正义链包含如SEQ ID NO:389所示的核苷酸序列，所述反义链包含如SEQ ID NO:382所示的核苷酸序列；184) The sense strand includes the nucleotide sequence shown in SEQ ID NO:389, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:382;
    185)所述正义链包含如SEQ ID NO:389所示的核苷酸序列，所述反义链包含如SEQ ID NO:383所示的核苷酸序列；185) The sense strand includes the nucleotide sequence shown in SEQ ID NO:389, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:383;
    186)所述正义链包含如SEQ ID NO:389所示的核苷酸序列，所述反义链包含如SEQ ID NO:384所示的核苷酸序列；186) The sense strand includes the nucleotide sequence shown in SEQ ID NO:389, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:384;
    187)所述正义链包含如SEQ ID NO:390所示的核苷酸序列，所述反义链包含如SEQ ID NO:394所示的核苷酸序列；187) The sense strand includes the nucleotide sequence shown in SEQ ID NO:390, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:394;
    188)所述正义链包含如SEQ ID NO:390所示的核苷酸序列，所述反义链包含如SEQ ID NO:395所示的核苷酸序列；188) The sense strand includes the nucleotide sequence shown in SEQ ID NO:390, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:395;
    189)所述正义链包含如SEQ ID NO:391所示的核苷酸序列，所述反义链包含如SEQ ID NO:395所示的核苷酸序列；189) The sense strand includes the nucleotide sequence shown in SEQ ID NO:391, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:395;
    190)所述正义链包含如SEQ ID NO:390所示的核苷酸序列，所述反义链包含如SEQ ID NO:396所示的核苷酸序列；190) The sense strand includes the nucleotide sequence shown in SEQ ID NO:390, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:396;
    191)所述正义链包含如SEQ ID NO:390所示的核苷酸序列，所述反义链包含如SEQ ID NO:397所示的核苷酸序列；191) The sense strand includes the nucleotide sequence shown in SEQ ID NO:390, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:397;
    192)所述正义链包含如SEQ ID NO:392所示的核苷酸序列，所述反义链包含如SEQ ID NO:398所示的核苷酸序列；192) The sense strand includes the nucleotide sequence shown in SEQ ID NO:392, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:398;
    193)所述正义链包含如SEQ ID NO:392所示的核苷酸序列，所述反义链包含如SEQ ID NO:399所示的核苷酸序列；193) The sense strand includes the nucleotide sequence shown in SEQ ID NO:392, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:399;
    194)所述正义链包含如SEQ ID NO:393所示的核苷酸序列，所述反义链包含如SEQ ID NO:399所示的核苷酸序列；194) The sense strand includes the nucleotide sequence shown in SEQ ID NO:393, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:399;
    195)所述正义链包含如SEQ ID NO:392所示的核苷酸序列，所述反义链包含如SEQ ID NO:400所示的核苷酸序列；195) The sense strand includes the nucleotide sequence shown in SEQ ID NO:392, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:400;
    196)所述正义链包含如SEQ ID NO:392所示的核苷酸序列，所述反义链包含如SEQ ID NO:401所示的核苷酸序列；196) The sense strand includes the nucleotide sequence shown in SEQ ID NO:392, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:401;
    197)所述正义链包含如SEQ ID NO:226所示的核苷酸序列，所述反义链包含如SEQ ID NO:377所示的核苷酸序列；197) The sense strand includes the nucleotide sequence shown in SEQ ID NO:226, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:377;
    198)所述正义链包含如SEQ ID NO:226所示的核苷酸序列，所述反义链包含如SEQ ID NO:378所示的核苷酸序列；198) The sense strand includes the nucleotide sequence shown in SEQ ID NO:226, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:378;
    199)所述正义链包含如SEQ ID NO:226所示的核苷酸序列，所述反义链包含如SEQ ID NO:402所示的核苷酸序列；199) The sense strand includes the nucleotide sequence shown in SEQ ID NO:226, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:402;
    200)所述正义链包含如SEQ ID NO:226所示的核苷酸序列，所述反义链包含如SEQ ID NO:403所示的核苷酸序列。 200) The sense strand includes the nucleotide sequence shown in SEQ ID NO:226, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:403.
  29. 一种双链核糖核酸缀合物，其中，所述双链核糖核酸缀合物包括如权利要求1-14任一项所述的双链核糖核酸，或如权利要求15-28任一项所述的双链核糖核酸修饰物；以及，缀合连接于所述双链核糖核酸或所述双链核糖核酸修饰物的缀合基团。A double-stranded ribonucleic acid conjugate, wherein the double-stranded ribonucleic acid conjugate includes the double-stranded ribonucleic acid as described in any one of claims 1-14, or as described in any one of claims 15-28 The double-stranded ribonucleic acid modification; and, a conjugation group connected to the double-stranded ribonucleic acid or the double-stranded ribonucleic acid modification.
  30. 根据权利要求29所述的双链核糖核酸缀合物，其中，所述缀合基团具有如下所示结构：
    The double-stranded ribonucleic acid conjugate according to claim 29, wherein the conjugation group has the structure shown below:
  31. 根据权利要求29或30所述的双链核糖核酸缀合物，其中，所述缀合基团连接于正义链的3’末端。The double-stranded ribonucleic acid conjugate according to claim 29 or 30, wherein the conjugation group is connected to the 3' end of the sense strand.
  32. 根据权利要求31所述的双链核糖核酸缀合物，其中，所述缀合基团通过磷酸二酯键与正义链的3’末端缀合连接；The double-stranded ribonucleic acid conjugate according to claim 31, wherein the conjugation group is conjugated to the 3′ end of the sense strand through a phosphodiester bond;
    优选地，所述双链核糖核酸缀合物的正义链与反义链互补形成所述双链核糖核酸缀合物的双链区，且所述正义链的3’末端形成平末端，所述反义链的3’末端具有1-2个延伸出所述双链区的突出的核苷酸；Preferably, the sense strand and the antisense strand of the double-stranded ribonucleic acid conjugate are complementary to form a double-stranded region of the double-stranded ribonucleic acid conjugate, and the 3' end of the sense strand forms a blunt end, and the The 3' end of the antisense strand has 1-2 protruding nucleotides extending out of the double-stranded region;
    或者，or,
    所述双链核糖核酸缀合物的正义链与反义链互补形成所述双链核糖核酸缀合物的双链区，且所述正义链的3’末端形成平末端，所述反义链的3’末端形成平末端。The sense strand and the antisense strand of the double-stranded ribonucleic acid conjugate are complementary to form the double-stranded region of the double-stranded ribonucleic acid conjugate, and the 3' end of the sense strand forms a blunt end, and the antisense strand The 3' end forms a blunt end.
  33. 根据权利要求29-32任一项所述的双链核糖核酸缀合物，其中，所述双链核糖核酸缀合物具有如下所示结构：
    The double-stranded ribonucleic acid conjugate according to any one of claims 29-32, wherein the double-stranded ribonucleic acid conjugate has the structure shown below:
    其中，双螺旋结构为双链核糖核酸或双链核糖核酸修饰物。Among them, the double helix structure is double-stranded ribonucleic acid or double-stranded ribonucleic acid modification.
  34. 根据权利要求29-33任一项所述的双链核糖核酸缀合物，其中，所述双链核糖核酸缀合物为siRNA缀合物。The double-stranded ribonucleic acid conjugate according to any one of claims 29-33, wherein the double-stranded ribonucleic acid conjugate is an siRNA conjugate.
  35. 根据权利要求29-34任一项所述的双链核糖核酸缀合物，其中，所述双链核糖核酸缀合物是用于抑制ANGPTL3基因表达的siRNA缀合物。The double-stranded ribonucleic acid conjugate according to any one of claims 29 to 34, wherein the double-stranded ribonucleic acid conjugate is an siRNA conjugate for inhibiting ANGPTL3 gene expression.
  36. 根据权利要求29-35任一项所述的双链核糖核酸缀合物，其中，所述双链核糖核酸缀合物由表1所示的任意一种siRNA与缀合基团连接形成，或者，所述双链核糖核酸缀合物由表1-1所示的任意一种siRNA与缀合基团连接形成，或者，所述双链核糖核酸缀合物由表2所示的任意一种siRNA修饰物与缀合基团连接形成；The double-stranded ribonucleic acid conjugate according to any one of claims 29-35, wherein the double-stranded ribonucleic acid conjugate is formed by connecting any siRNA shown in Table 1 with a conjugating group, or , the double-stranded ribonucleic acid conjugate is formed by connecting any siRNA shown in Table 1-1 with a conjugation group, or the double-stranded ribonucleic acid conjugate is formed by any one of the siRNA shown in Table 2 The siRNA modification is formed by connecting with the conjugation group;
    优选地，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID  NO:255所示的核苷酸序列，所述反义链包含如SEQ ID NO:241所示的核苷酸序列；Preferably, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes SEQ ID The nucleotide sequence shown in NO:255, the antisense strand includes the nucleotide sequence shown in SEQ ID NO:241;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:256所示的核苷酸序列，所述反义链包含如SEQ ID NO:247所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:256, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:247 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:257所示的核苷酸序列，所述反义链包含如SEQ ID NO:253所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:257, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:253 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:256所示的核苷酸序列，所述反义链包含如SEQ ID NO:377所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 256, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 377 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:256所示的核苷酸序列，所述反义链包含如SEQ ID NO:378所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 256, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 378 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:256所示的核苷酸序列，所述反义链包含如SEQ ID NO:379所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 256, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 379 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:256所示的核苷酸序列，所述反义链包含如SEQ ID NO:380所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 256, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 380 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:257所示的核苷酸序列，所述反义链包含如SEQ ID NO:366所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 257, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 366 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:257所示的核苷酸序列，所述反义链包含如SEQ ID NO:367所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 257, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 367 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:257所示的核苷酸序列，所述反义链包含如SEQ ID NO:368所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 257, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 368 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:257所示的核苷酸序列，所述反义链包含如SEQ ID NO:369所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 257, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 369 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:374所示的核苷酸序列，所述反义链包含如SEQ ID NO:362所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:374, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:362 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:374所示的核苷酸序列，所述反义链包含如SEQ ID NO:363所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:374, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:363 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:374所示的核苷酸序列，所述反义链包含如SEQ ID NO:364所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:374, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:364 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:374所示的核苷酸序列，所述反义链包含如SEQ ID NO:365所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:374, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:365 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:375所示的核苷酸序列，所述反义链包含如SEQ ID NO:370所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:375, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:370 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:375所示的核苷酸序列，所述反义链包含如SEQ ID NO:371所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:375, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:371 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:375所示的核苷酸序列，所述反义链包含如SEQ ID NO:372所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:375, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:372 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:375所示的核苷酸序列，所述反义链包含如SEQ ID NO:373所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:375, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:373 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:376所示的核苷酸序列，所述反义链包含如SEQ ID NO:381所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:376, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:381 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:376所示的核苷酸序列，所述反义链包含如SEQ ID NO:382所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:376, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:382 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:376所示的核苷酸序列，所述反义链包含如SEQ ID NO:383所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:376, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:383 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:376所示的核苷酸序列，所述反义链包含如SEQ ID NO:384所示的核苷酸序列； Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:376, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:384. The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:374所示的核苷酸序列，所述反义链包含如SEQ ID NO:252所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 374, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 252 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:375所示的核苷酸序列，所述反义链包含如SEQ ID NO:254所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 375, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 254 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:404所示的核苷酸序列，所述反义链包含如SEQ ID NO:394所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 404, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 394 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:404所示的核苷酸序列，所述反义链包含如SEQ ID NO:395所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 404, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 395 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:404所示的核苷酸序列，所述反义链包含如SEQ ID NO:396所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 404, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 396 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:404所示的核苷酸序列，所述反义链包含如SEQ ID NO:397所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 404, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 397 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:405所示的核苷酸序列，所述反义链包含如SEQ ID NO:398所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 405, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 398 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:405所示的核苷酸序列，所述反义链包含如SEQ ID NO:399所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO: 405, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO: 399 The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:405所示的核苷酸序列，所述反义链包含如SEQ ID NO:400所示的核苷酸序列；Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:405, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:400. The nucleotide sequence shown;
    或者，所述双链核糖核酸缀合物中，连接缀合物基团的正义链包含如SEQ ID NO:405所示的核苷酸序列，所述反义链包含如SEQ ID NO:401所示的核苷酸序列。Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the nucleotide sequence shown in SEQ ID NO:405, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:401 The nucleotide sequence shown.
  37. 一种药物组合物，其中，所述药物组合物包括如下至少一项：如权利要求1-14任一项所述的双链核糖核酸，如权利要求15-28任一项所述的双链核糖核酸修饰物，如权利要求29-36任一项所述的双链核糖核酸缀合物。A pharmaceutical composition, wherein the pharmaceutical composition includes at least one of the following: the double-stranded ribonucleic acid according to any one of claims 1-14, the double-stranded ribonucleic acid according to any one of claims 15-28 Ribonucleic acid modification, such as the double-stranded ribonucleic acid conjugate according to any one of claims 29-36.
  38. 根据权利要求37所述的药物组合物，其中，所述药物组合物还包括一种或多种药学上可接收的载体。The pharmaceutical composition according to claim 37, wherein the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers.
  39. 根据权利要求1-14任一项所述的双链核糖核酸，根据权利要求15-28任一项所述的双链核糖核酸修饰物，根据权利要求29-36任一项所述的双链核糖核酸缀合物，或根据权利要求37-38任一项所述的药物组合物在如下至少一项中的用途：The double-stranded ribonucleic acid according to any one of claims 1-14, the double-stranded ribonucleic acid modification according to any one of claims 15-28, the double-stranded ribonucleic acid according to any one of claims 29-36 Ribonucleic acid conjugate, or the use of the pharmaceutical composition according to any one of claims 37-38 in at least one of the following:
    (1)抑制ANGPTL3基因表达，或制备用于抑制ANGPTL3基因表达的药物；(1) Inhibit ANGPTL3 gene expression, or prepare drugs for inhibiting ANGPTL3 gene expression;
    (2)用于预防或治疗与ANGPTL3基因异常表达相关的疾病，或制备用于预防或治疗与ANGPTL3基因异常表达相关的疾病的药物；(2) For preventing or treating diseases related to abnormal expression of ANGPTL3 gene, or preparing drugs for preventing or treating diseases related to abnormal expression of ANGPTL3 gene;
    (3)用于治疗患有将受益于ANGPTL3基因表达降低的疾病的受试者，或制备用于治疗患有将受益于ANGPTL3基因表达降低的疾病的受试者的药物。(3) For use in treating subjects suffering from a disease that would benefit from a reduction in ANGPTL3 gene expression, or to prepare a medicament for treating subjects suffering from a disease that would benefit from a reduction in ANGPTL3 gene expression.
  40. 根据权利要求39所述的用途，其中，所述与ANGPTL3基因异常表达相关的疾病选自与脂代谢相关的疾病；The use according to claim 39, wherein the disease related to abnormal expression of ANGPTL3 gene is selected from diseases related to lipid metabolism;
    可选地，所述与脂代谢相关的疾病选自如下疾病组成的组：Optionally, the disease related to lipid metabolism is selected from the group consisting of the following diseases:
    乳糜微粒血症综合征、2型糖尿病、家族性部分脂肪营养不良、高胆固醇血症、家族性高胆固醇血症、高甘油三酯血症、非酒精性脂肪肝、动脉粥样硬化、高甘油三酯血症、膝盖损伤及骨关节炎、血脂异常。Chylomicronemia syndrome, type 2 diabetes, familial partial lipodystrophy, hypercholesterolemia, familial hypercholesterolemia, hypertriglyceridemia, non-alcoholic fatty liver disease, atherosclerosis, hyperglycemia Triesteremia, knee injury and osteoarthritis, dyslipidemia.
  41. 一种用于抑制细胞内ANGPTL3基因表达的方法，其中，所述方法包括将所述细胞与根据权利要求1-14任一项所述的双链核糖核酸，根据权利要求15-28任一项所述的双链核糖核酸修饰物，根据权利要求29-36任一项所述的双链核糖核酸缀合物，或根据权利要求37-38任一项所述的药物组合物接触。A method for inhibiting intracellular ANGPTL3 gene expression, wherein the method includes combining the cell with the double-stranded ribonucleic acid according to any one of claims 1-14, according to any one of claims 15-28 The double-stranded ribonucleic acid modification is a double-stranded ribonucleic acid conjugate according to any one of claims 29-36, or a pharmaceutical composition according to any one of claims 37-38.
  42. 根据权利要求41所述的方法，其中，所述细胞为体内细胞或体外细胞。The method of claim 41, wherein the cells are in vivo cells or in vitro cells.
  43. 根据权利要求41或42所述的方法，其中，所述细胞在受试者体内。 The method of claim 41 or 42, wherein the cells are in the subject.
  44. 根据权利要求43所述的方法，其中，所述受试者为哺乳动物，优选为人。The method of claim 43, wherein the subject is a mammal, preferably a human.
  45. 根据权利要求43或44所述的方法，其中，所述受试者具有如下至少一种特性：The method of claim 43 or 44, wherein the subject has at least one of the following characteristics:
    体内ANGPTL3基因异常表达，更具体地为ANGPTL3基因异常高表达；Abnormal expression of ANGPTL3 gene in vivo, more specifically abnormal high expression of ANGPTL3 gene;
    患有与ANGPTL3基因异常表达相关的疾病；Suffering from diseases related to abnormal expression of ANGPTL3 gene;
    患有将受益于ANGPTL3基因表达降低的疾病。 People with diseases that would benefit from reduced ANGPTL3 gene expression.
PCT/CN2023/108634 2022-08-04 2023-07-21 Double-stranded ribonucleic acid for inhibiting angptl3 gene expression, and modifier, conjugate, and use thereof WO2024027518A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202210935615 2022-08-04
CN202210935615.8 2022-08-04

Publications (1)

Publication Number Publication Date
WO2024027518A1 true WO2024027518A1 (en) 2024-02-08

Family

ID=88529420

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2023/108634 WO2024027518A1 (en) 2022-08-04 2023-07-21 Double-stranded ribonucleic acid for inhibiting angptl3 gene expression, and modifier, conjugate, and use thereof

Country Status (2)

Country Link
CN (1) CN116987696A (en)
WO (1) WO2024027518A1 (en)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107743522A (en) * 2015-04-13 2018-02-27 阿尔尼拉姆医药品有限公司 Class angiogenin 3（ANGPTL3）IRNA compositions and its application method
CN110945130A (en) * 2017-12-01 2020-03-31 苏州瑞博生物技术有限公司 Nucleic acid, composition containing nucleic acid, conjugate, preparation method and application
CN111343994A (en) * 2017-09-14 2020-06-26 箭头药业股份有限公司 RNAi agents and compositions for inhibiting expression of angiopoietin-like 3(ANGPTL3) and methods of use
CN111973617A (en) * 2019-05-23 2020-11-24 苏州瑞博生物技术股份有限公司 Nucleic acid, pharmaceutical composition and conjugate, preparation method and application
WO2022068923A1 (en) * 2020-09-30 2022-04-07 纳肽得(青岛)生物医药有限公司 Sirna of angiopoietin-like 3 (angptl3) and use thereof
WO2022079222A1 (en) * 2020-10-16 2022-04-21 Sanofi Novel rna compositions and methods for inhibiting angptl3

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107743522A (en) * 2015-04-13 2018-02-27 阿尔尼拉姆医药品有限公司 Class angiogenin 3（ANGPTL3）IRNA compositions and its application method
CN111343994A (en) * 2017-09-14 2020-06-26 箭头药业股份有限公司 RNAi agents and compositions for inhibiting expression of angiopoietin-like 3(ANGPTL3) and methods of use
CN110945130A (en) * 2017-12-01 2020-03-31 苏州瑞博生物技术有限公司 Nucleic acid, composition containing nucleic acid, conjugate, preparation method and application
CN111973617A (en) * 2019-05-23 2020-11-24 苏州瑞博生物技术股份有限公司 Nucleic acid, pharmaceutical composition and conjugate, preparation method and application
WO2022068923A1 (en) * 2020-09-30 2022-04-07 纳肽得(青岛)生物医药有限公司 Sirna of angiopoietin-like 3 (angptl3) and use thereof
WO2022079222A1 (en) * 2020-10-16 2022-04-21 Sanofi Novel rna compositions and methods for inhibiting angptl3

Also Published As

Publication number Publication date
CN116987696A (en) 2023-11-03

Similar Documents

Publication Publication Date Title
EP3315608B1 (en) Sirna, pharmaceutical composition and conjugate which contain sirna, and uses thereof
JP2022532169A (en) Oligonucleotide composition and its usage
TW202126810A (en) Oligonucleotide compositions and methods of use thereof
JP2020534253A (en) Oligonucleotide composition and its method
CN106983768A (en) Composition and method for adjusting SMN2 montages in object
WO2020233655A1 (en) Nucleic acid, pharmaceutical composition, conjugate, preparation method, and use
IL194419A (en) Dsrna for inhibiting the expression of human eg5 gene in a cell, a pharmaceutical composition comprising same, method and vector
BR112015024764B1 (en) MODIFIED TGF-BETA OLIGONUCLEOTIDES, THEIR USE AND PHARMACEUTICAL COMPOSITIONS INCLUDING THEM
KR20210110310A (en) Oligomeric Nucleic Acid Molecules and Their Uses
CN117070517A (en) siRNA for inhibiting expression of angiotensinogen, conjugate, pharmaceutical composition and application thereof
WO2024008114A1 (en) Sirna for inhibiting complement factor b expression, pharmaceutical composition and conjugate thereof, and use thereof
WO2024027518A1 (en) Double-stranded ribonucleic acid for inhibiting angptl3 gene expression, and modifier, conjugate, and use thereof
WO2022143531A1 (en) Nucleic acid, pharmaceutical composition and sirna conjugate containing the nucleic acid, preparation method therefor, and use thereof
CN115141844A (en) RNA plasmid delivery system and application thereof
WO2020233651A1 (en) Nucleic acid, pharmaceutical composition, conjugate, preparation method, and use
WO2024061202A1 (en) Double-stranded ribonucleic acid for inhibiting hmgb1 gene expression, and modifier, conjugate and use thereof
WO2023227070A1 (en) Double-stranded ribonucleic acid for inhibiting c3 gene expression and modifier, conjugate, and use thereof
WO2024008158A1 (en) Double-stranded ribonucleic acid for inhibiting c5 gene expression, and a modifier, a conjugate and use thereof
WO2023213284A1 (en) Nucleic acid, composition and conjugate comprising same, preparation method therefor, and use thereof
CN117384902A (en) Double-stranded ribonucleic acid for inhibiting CYP27A1 gene expression, and modification, conjugate and application thereof
WO2023208109A9 (en) Sirna for inhibiting hsd17b13 expression, conjugate and pharmaceutical composition thereof, and use thereof
WO2024046297A1 (en) Sirna for inhibiting expression of asialoglycoprotein receptor gene and conjugate, pharmaceutical composition, and use thereof
CN116497024A (en) Nucleic acid and composition, preparation method and application thereof
CN116497027A (en) Nucleic acid, pharmaceutical composition and conjugate and use
CN117660449A (en) Nucleic acid, pharmaceutical composition and conjugate and use

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23849226

Country of ref document: EP

Kind code of ref document: A1