WO2023227070A1

WO2023227070A1 - Double-stranded ribonucleic acid for inhibiting c3 gene expression and modifier, conjugate, and use thereof

Info

Publication number: WO2023227070A1
Application number: PCT/CN2023/096337
Authority: WO
Inventors: 王书成; 黄河; 刘薇; 王岩; 林国良; 汪小君; 荣梅
Original assignee: Beijing Winsunny Pharmaceutical Co Ltd
Current assignee: Beijing Foyou Pharmaceutical Co Ltd
Priority date: 2022-05-27
Filing date: 2023-05-25
Publication date: 2023-11-30
Anticipated expiration: 2024-11-27
Also published as: CN116622710A

Abstract

The present disclosure relates to a double-stranded ribonucleic acid for inhibiting C3 gene expression, a modifier, a conjugate, and a 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 a use for inhibiting C3 gene expression, as well as a method for inhibiting C3 gene expression in cells. The provided double-stranded ribonucleic acid can be combined in cells to form an RNA-induced silencing complex (RISC), and cleaves complement C3 gene-transcribed mRNA, efficiently and specifically inhibiting the expression of the complement C3 gene, same is used for treating an improper complement system activation-mediated disease, and has important application prospects in clinical treatment of disease.

Description

Double-stranded ribonucleic acid for inhibiting C3 gene expression and its modifications, conjugates and uses

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C3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C3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 C3 gene expression, and Methods for inhibiting intracellular C3 gene expression.

Background technique

补体(Complement，C)是存在于人和动物血清与组织液中的一组不耐热、经活化后具有酶活性、可介导免疫应答和炎症反应的蛋白质。补体系统(Complement system)被激活后，介导一系列的细胞反应，如细胞溶解、调理吞噬(抗原抗体结合)、炎症反应、清除免疫复合物等。补体系统主要由固有成分、调节蛋白及受体等30余种糖蛋白组成，这些蛋白质以血液中可溶性蛋白存在或以膜相关蛋白存在。补体的激活引起依序的酶促反应级联(称为补体激活途径)，从而导致有效的过敏毒素C3a和C5a的形成，这些过敏毒素诱发从化学吸引到细胞凋亡范围内的多种生理学反应。Complement (Complement, C) is a group of heat-labile proteins that exist in the serum and tissue fluid of humans and animals, have enzymatic activity after activation, and can mediate immune responses and inflammatory responses. After the complement system is activated, it mediates a series of cellular reactions, such as cell lysis, opsonophagocytosis (antigen-antibody binding), inflammatory response, clearance of immune complexes, etc. The complement system is mainly composed of more than 30 types of glycoproteins such as intrinsic components, regulatory proteins and receptors. These proteins exist as soluble proteins in the blood or as membrane-associated proteins. Activation of complement causes a sequential cascade of enzymatic reactions (called the complement activation pathway), leading to the formation of the potent anaphylatoxins C3a and C5a, which induce a variety of physiological responses ranging from chemical attraction to apoptosis .

已知补体激活通过三种不同途径发生：旁路途径、经典途径和凝集素途径，涉及主要作为非活性酶原存在的蛋白质，然后这些非活性酶原依序地切割并且激活。三条途径起始物各异，但相互交叉并具有共同的末端通路。经典途径的激活始于C1q与免疫复合物的Fc段结合，依赖特异性抗体的产生，故在感染后期或恢复期发挥作用。旁路途径则由细菌、真菌或病毒感染细胞等直接为自发产生的C3b提供反应表面启动激活，无需抗体存在，故在抗体产生前的感染早期或初次感染发挥作用。凝集素途径也无需抗体参与即可激活补体，其激活物质非常广泛，主要是MBL或纤维胶原素直接识别病原微生物表面的N氨基半乳糖或甘露糖，除识别机制不同于经典途径外，其后续活化途径基本相同，对经典途径和旁路途径具有交叉促进作用。三条补体激活途径在C3这一环节汇合进入共有的末端通路,形成C5转化酶，激活C5裂解成C5a和C5b。C5b结合于细胞表面，依次与C6、C7、C8和12～15个C9分子结合形成C5b6789n，即膜攻击复合物(MAC)，介导细胞溶解效应。Complement activation is known to occur through three different pathways: the alternative pathway, the classical pathway, and the lectin pathway, involving proteins that exist primarily as inactive zymogens, which are then sequentially cleaved and activated. The three pathways have different starting materials, but they intersect with each other and have a common terminal pathway. The activation of the classical pathway begins with the binding of C1q to the Fc segment of the immune complex and relies on the production of specific antibodies, so it plays a role in the late infection or recovery period. The alternative pathway directly provides a reaction surface for spontaneously produced C3b to initiate activation when infected cells by bacteria, fungi or viruses do not require the presence of antibodies, so it plays a role in the early stages of infection or the first infection before antibodies are produced. The lectin pathway can also activate complement without the involvement of antibodies. Its activation substances are very wide, mainly MBL or fibrillar collagen that directly recognize N-galactosamine or mannose on the surface of pathogenic microorganisms. In addition to the recognition mechanism being different from the classic pathway, its subsequent The activation pathways are basically the same, with cross-promoting effects on the classical and alternative pathways. The three complement activation pathways merge into a common terminal pathway at the C3 link to form C5 convertase, which activates C5 to cleave into C5a and C5b. C5b binds to the cell surface and sequentially combines with C6, C7, C8 and 12 to 15 C9 molecules to form C5b6789n, the membrane attack complex (MAC), which mediates the cell lysis effect.

补体系统的不适当的激活是在许多疾病中的增殖和/或起始病变的原因，这些疾病包括例如，阵发性睡眠性血红蛋白尿症、非典型溶血性尿毒症综合征、类风湿性关节炎、慢性肾病、缺血再灌注损伤以及神经退行性疾病。Inappropriate activation of the complement system is responsible for proliferation and/or initiating pathology in many diseases, including, for example, paroxysmal nocturnal hemoglobinuria, atypical hemolytic uremic syndrome, rheumatoid arthritis inflammation, chronic kidney disease, ischemia-reperfusion injury, and neurodegenerative diseases.

开发C3补体抑制剂是一项有前景的治疗方法，2021年5月，美国FDA批准Apellis Pharmaceuticals公司的Empaveli(为合成环状肽)上市，用于治疗初治阵发性睡眠性血红蛋白尿(PNH)患者，这是首个且唯一个用于治疗PNH的C3靶向疗法。因此，开发新型补体C3抑制剂拥有巨大的临床价值和市场前景。The development of C3 complement inhibitors is a promising treatment method. In May 2021, the US FDA approved Apellis Pharmaceuticals' Empaveli (a synthetic cyclic peptide) for the treatment of newly treated paroxysmal nocturnal hemoglobinuria (PNH). ) patients, this is the first and only C3-targeted therapy for the treatment of PNH. Therefore, the development of new complement C3 inhibitors has great clinical value and market prospects.

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C3补体抑制剂，用于治疗补体系统的不适当的激活介导的疾病。本公开旨在提供一系列用于抑制补体C3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C3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, there is a need to develop more C3 complement inhibitors for the treatment of diseases mediated by inappropriate activation of the complement system. The present disclosure aims to provide a series of double-stranded ribonucleic acid, double-stranded ribonucleic acid modifications, double-stranded ribonucleic acid conjugates and pharmaceutical compositions for inhibiting complement C3 gene expression, which can inhibit C3 gene expression and be used in clinical disease treatment. It has 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:8-34、SEQ ID NO:364任一项所示的核苷酸序列，所述正义链包含如SEQ ID NO:8-34、SEQ ID NO:364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A，所述反义链包含如SEQ ID NO:8-34、SEQ ID NO:364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反向互补和/或基本上反向互补的序列B。[2]. The double-stranded ribonucleic acid according to [1], wherein the target sequence is selected from the nucleotide sequence shown in any one of SEQ ID NO:8-34 and SEQ ID NO:364, and the The sense strand includes a sequence A consisting of at least 15 consecutive nucleotides in the nucleotide sequence shown in any one of SEQ ID NO: 8-34 and SEQ ID NO: 364, and the antisense strand includes a sequence A as shown in any one of SEQ ID NO: 8-34 and SEQ ID NO: 364 :8-34. A reverse complementary and/or substantially reverse complementary sequence B consisting of at least 15 consecutive nucleotides in the nucleotide sequence shown in any one of SEQ ID NO:364.

[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SEQ ID NO:8-34、SEQ ID NO:364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 the nucleotide sequence shown in any one of SEQ ID NO: 8-34 and SEQ ID NO: 364 Sequence A consisting of 15-28 consecutive nucleotides in the sequence 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:8-34、SEQ ID NO:364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反向互补和/或基本上反向互补的序列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 the same as that shown in any one of SEQ ID NO: 8-34 and SEQ ID NO: 364. Sequence B consisting of 15-28 consecutive nucleotides in the nucleotide sequence is reverse complementary and/or substantially reverse complementary, preferably 19-25 consecutive nucleotides, more preferably 19-23 consecutive nucleotides acid, more preferably 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:35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；1) 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:131;

2)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:132所示的核苷酸序列；2) 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:132;

3)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:133所示的核苷酸序列；3) 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:133;

4)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:134所示的核苷酸序列；4) 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:134;

5)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:135所示的核苷酸序列；5) 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:135;

6)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:136所示的核苷酸序列； 6) 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:136;

7)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:137所示的核苷酸序列；7) 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:137;

8)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:138所示的核苷酸序列；8) 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:138;

9)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:139所示的核苷酸序列；9) 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:139;

10)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:140所示的核苷酸序列；10) 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:140;

11)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:141所示的核苷酸序列；11) 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:141;

12)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:142所示的核苷酸序列；12) 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:142;

13)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:143所示的核苷酸序列；13) 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:143;

14)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:144所示的核苷酸序列；14) 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:144;

15)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:145所示的核苷酸序列；15) 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:145;

16)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:146所示的核苷酸序列；16) 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:146;

17)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:147所示的核苷酸序列；17) 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:147;

18)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:148所示的核苷酸序列；18) 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:148;

19)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:149所示的核苷酸序列；19) 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:149;

20)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:150所示的核苷酸序列；20) 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:150;

21)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:151所示的核苷酸序列；21) 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:151;

22)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:152所示的核苷酸序列；22) 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:152;

23)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:153所示的核苷酸序列；23) 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:153;

24)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:154所示的核苷酸序列；24) 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:154;

25)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:155所示的核苷酸序列；25) 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:155;

26)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:156所示的核苷酸序列；26) 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:156;

27)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:157所示的核苷酸序列；27) 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:157;

28)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:158所示的核苷酸序列；28) 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:158;

29)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:159所示的核苷酸序列；29) 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:63 The nucleotide sequence shown in NO:159;

30)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:160所示的核苷酸序列；30) 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:160;

31)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:161所示的核苷酸序列；31) 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:161;

32)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:162所示的核苷酸序列；32) 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:162;

33)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:163所示的核苷酸序列；33) 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: 163;

34)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:164所示的核苷酸序列；34) 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:164;

35)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:165所示的核苷酸序列；35) 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:165;

36)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:166所示的核苷酸序列；36) 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:166;

37)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:167所示的核苷酸序列；37) 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:167;

38)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:168所示的核苷酸序列；38) 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:168;

39)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:169所示的核苷酸序列；39) 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:169;

40)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:170所示的核苷酸序列；40) 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:170;

41)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:171所示的核苷酸序列；41) 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:171;

42)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:172所示的核苷酸序列；42) 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:172;

43)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:173所示的核苷酸序列；43) 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:173;

44)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:174所示的核苷酸序列；44) 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:174;

45)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:175所示的核苷酸序列；45) 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:175;

46)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:176所示的核苷酸序列；46) 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:176;

47)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:177所示的核苷酸序列；47) 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:177;

48)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:178所示的核苷酸序列；48) 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:178;

49)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:179所示的核苷酸序列；49) 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:179;

50)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:180所示的核苷酸序列；50) 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:180;

51)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:181所示的核苷酸序列； 51) 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:181;

52)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:182所示的核苷酸序列；52) 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:182;

53)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:183所示的核苷酸序列；53) 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:183;

54)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:184所示的核苷酸序列；54) 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:184;

55)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:185所示的核苷酸序列；55) 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:185;

56)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:186所示的核苷酸序列；56) 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:186;

57)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:187所示的核苷酸序列；57) 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:187;

58)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:188所示的核苷酸序列；58) 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:188;

59)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:189所示的核苷酸序列；59) 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:189;

60)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:190所示的核苷酸序列；60) 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:190;

61)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:191所示的核苷酸序列；61) 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:191;

62)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:192所示的核苷酸序列；62) 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:192;

63)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:193所示的核苷酸序列；63) 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:193;

64)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:194所示的核苷酸序列；64) 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:194;

65)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:195所示的核苷酸序列；65) 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:195;

66)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:196所示的核苷酸序列；66) 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:196;

67)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:197所示的核苷酸序列；67) 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:197;

68)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:198所示的核苷酸序列；68) 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:198;

69)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:199所示的核苷酸序列；69) 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:199;

70)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:200所示的核苷酸序列；70) 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:200;

71)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:201所示的核苷酸序列；71) 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:201;

72)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:202所示的核苷酸序列；72) 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:202;

73)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:203所示的核苷酸序列；73) 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:203;

74)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:204所示的核苷酸序列；74) 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. The nucleotide sequence shown in NO:204;

75)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:205所示的核苷酸序列；75) 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:205;

76)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:206所示的核苷酸序列；76) 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:206;

77)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:207所示的核苷酸序列；77) 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:207;

78)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:208所示的核苷酸序列；78) 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:208;

79)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:209所示的核苷酸序列；79) 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:209;

80)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:210所示的核苷酸序列；80) 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:210;

81)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:211所示的核苷酸序列；81) 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:211;

82)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:212所示的核苷酸序列；82) 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:212;

83)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:213所示的核苷酸序列；83) 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:213;

84)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:214所示的核苷酸序列；84) 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:214;

85)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:215所示的核苷酸序列；85) 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:215;

86)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:216所示的核苷酸序列；86) 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:216;

87)所述正义链包含如SEQ ID NO:121所示的核苷酸序列，所述反义链包含如SEQ ID NO:217所示的核苷酸序列；87) The sense strand includes the nucleotide sequence shown in SEQ ID NO:121, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:217;

88)所述正义链包含如SEQ ID NO:122所示的核苷酸序列，所述反义链包含如SEQ ID NO:218所示的核苷酸序列；88) The sense strand includes the nucleotide sequence shown in SEQ ID NO:122, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:218;

89)所述正义链包含如SEQ ID NO:123所示的核苷酸序列，所述反义链包含如SEQ ID NO:219所示的核苷酸序列；89) The sense strand includes the nucleotide sequence shown in SEQ ID NO:123, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:219;

90)所述正义链包含如SEQ ID NO:124所示的核苷酸序列，所述反义链包含如SEQ ID NO:220所示的核苷酸序列；90) The sense strand includes the nucleotide sequence shown in SEQ ID NO:124, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:220;

91)所述正义链包含如SEQ ID NO:125所示的核苷酸序列，所述反义链包含如SEQ ID NO:221所示的核苷酸序列；91) The sense strand includes the nucleotide sequence shown in SEQ ID NO:125, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:221;

92)所述正义链包含如SEQ ID NO:126所示的核苷酸序列，所述反义链包含如SEQ ID NO:222所示的核苷酸序列；92) The sense strand includes the nucleotide sequence shown in SEQ ID NO:126, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:222;

93)所述正义链包含如SEQ ID NO:127所示的核苷酸序列，所述反义链包含如SEQ ID NO:223所示的核苷酸序列；93) The sense strand includes the nucleotide sequence shown in SEQ ID NO:127, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:223;

94)所述正义链包含如SEQ ID NO:128所示的核苷酸序列，所述反义链包含如SEQ ID NO:224所示的核苷酸序列；94) The sense strand includes the nucleotide sequence shown in SEQ ID NO:128, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:224;

95)所述正义链包含如SEQ ID NO:129所示的核苷酸序列，所述反义链包含如SEQ ID NO:225所示的核苷酸序列；95) The sense strand includes the nucleotide sequence shown in SEQ ID NO:129, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:225;

118)所述正义链包含如SEQ ID NO:328所示的核苷酸序列，所述反义链包含如SEQ ID NO:346所示的核苷酸序列； 118) The sense strand includes the nucleotide sequence shown in SEQ ID NO:328, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:346;

119)所述正义链包含如SEQ ID NO:329所示的核苷酸序列，所述反义链包含如SEQ ID NO:347所示的核苷酸序列；119) The sense strand includes the nucleotide sequence shown in SEQ ID NO:329, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:347;

120)所述正义链包含如SEQ ID NO:330所示的核苷酸序列，所述反义链包含如SEQ ID NO:348所示的核苷酸序列；120) The sense strand includes the nucleotide sequence shown in SEQ ID NO:330, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:348;

121)所述正义链包含如SEQ ID NO:331所示的核苷酸序列，所述反义链包含如SEQ ID NO:349所示的核苷酸序列；121) The sense strand includes the nucleotide sequence shown in SEQ ID NO:331, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:349;

122)所述正义链包含如SEQ ID NO:332所示的核苷酸序列，所述反义链包含如SEQ ID NO:350所示的核苷酸序列；122) The sense strand includes the nucleotide sequence shown in SEQ ID NO:332, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:350;

123)所述正义链包含如SEQ ID NO:333所示的核苷酸序列，所述反义链包含如SEQ ID NO:351所示的核苷酸序列；123) The sense strand includes the nucleotide sequence shown in SEQ ID NO:333, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:351;

124)所述正义链包含如SEQ ID NO:334所示的核苷酸序列，所述反义链包含如SEQ ID NO:352所示的核苷酸序列；124) The sense strand includes the nucleotide sequence shown in SEQ ID NO:334, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:352;

125)所述正义链包含如SEQ ID NO:335所示的核苷酸序列，所述反义链包含如SEQ ID NO:353所示的核苷酸序列；125) The sense strand includes the nucleotide sequence shown in SEQ ID NO:335, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:353;

126)所述正义链包含如SEQ ID NO:336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；126) The sense strand includes the nucleotide sequence shown in SEQ ID NO:336, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:354;

127)所述正义链包含如SEQ ID NO:337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；127) The sense strand includes the nucleotide sequence shown in SEQ ID NO:337, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:355;

128)所述正义链包含如SEQ ID NO:338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；128) The sense strand includes the nucleotide sequence shown in SEQ ID NO:338, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:356;

129)所述正义链包含如SEQ ID NO:339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；129) The sense strand includes the nucleotide sequence shown in SEQ ID NO:339, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:357;

130)所述正义链包含如SEQ ID NO:340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；130) The sense strand includes the nucleotide sequence shown in SEQ ID NO:340, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:358;

131)所述正义链包含如SEQ ID NO:341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；131) The sense strand includes the nucleotide sequence shown in SEQ ID NO:341, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:359;

132)所述正义链包含如SEQ ID NO:342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；132) The sense strand includes the nucleotide sequence shown in SEQ ID NO:342, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:360;

133)所述正义链包含如SEQ ID NO:343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；133) The sense strand includes the nucleotide sequence shown in SEQ ID NO:343, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:361;

134)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:362所示的核苷酸序列；134) 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:362;

135)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:363所示的核苷酸序列。135) 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:363.

[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 C3基因表达的siRNA。[14]. The double-stranded ribonucleic acid according to any one of [1]-[13], wherein the double-stranded ribonucleic acid is used to inhibit C3 gene expression siRNA.

[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 ';

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 with 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C3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 C3 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:

97)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:242所示的核苷酸序列；97) 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:242;

98)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:243所示的核苷酸序列； 98) 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:243;

99)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:244所示的核苷酸序列；99) 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:244;

100)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:245所示的核苷酸序列；100) 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:245;

101)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:246所示的核苷酸序列；101) 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:246;

102)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:247所示的核苷酸序列；102) 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:247;

103)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:248所示的核苷酸序列；103) 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:248;

104)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:249所示的核苷酸序列；104) 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:249;

105)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:250所示的核苷酸序列；105) 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:250;

106)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:251所示的核苷酸序列；106) 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:251;

107)所述正义链包含如SEQ ID NO:237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；107) The sense strand includes the nucleotide sequence shown in SEQ ID NO:237, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:252;

108)所述正义链包含如SEQ ID NO:238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；108) The sense strand includes the nucleotide sequence shown in SEQ ID NO:238, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:253;

109)所述正义链包含如SEQ ID NO:239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；109) The sense strand includes the nucleotide sequence shown in SEQ ID NO:239, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:254;

110)所述正义链包含如SEQ ID NO:240所示的核苷酸序列，所述反义链包含如SEQ ID NO:255所示的核苷酸序列；110) The sense strand includes the nucleotide sequence shown in SEQ ID NO:240, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:255;

111)所述正义链包含如SEQ ID NO:241所示的核苷酸序列，所述反义链包含如SEQ ID NO:256所示的核苷酸序列；111) The sense strand includes the nucleotide sequence shown in SEQ ID NO:241, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:256;

115)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:260所示的核苷酸序列；115) 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:260;

116)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:261所示的核苷酸序列；116) 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:261;

117)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:262所示的核苷酸序列；117) 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:262;

136)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:283所示的核苷酸序列；136) 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:283;

137)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:283所示的核苷酸序列；137) 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:283;

138)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:284所示的核苷酸序列；138) 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:284;

139)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:285所示的核苷酸序列；139) 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:285;

140)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:286所示的核苷酸序列；140) 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:286;

141)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:287所示的核苷酸序列；141) 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:287;

142)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:287所示的核苷酸序列；142) The sense strand comprises the nucleotide sequence shown in SEQ ID NO:267, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:267 The nucleotide sequence shown in NO:287;

143)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:288所示的核苷酸序列；143) 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:288;

144)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:289所示的核苷酸序列；144) 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:289;

145)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:290所示的核苷酸序列；145) 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:290;

146)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:291所示的核苷酸序列；146) 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:291;

147)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:291所示的核苷酸序列；147) 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:291;

148)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:292所示的核苷酸序列；148) 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:292;

149)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:293所示的核苷酸序列；149) 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:293;

150)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:294所示的核苷酸序列；150) 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:294;

151)所述正义链包含如SEQ ID NO:238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；151) The sense strand includes the nucleotide sequence shown in SEQ ID NO:238, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:295;

152)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:295所示的核苷酸序列；152) 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:295;

153)所述正义链包含如SEQ ID NO:238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；153) The sense strand includes the nucleotide sequence shown in SEQ ID NO:238, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:296;

154)所述正义链包含如SEQ ID NO:238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；154) The sense strand includes the nucleotide sequence shown in SEQ ID NO:238, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:297;

155)所述正义链包含如SEQ ID NO:238所示的核苷酸序列，所述反义链包含如SEQ ID NO:298所示的核苷酸序列；155) The sense strand includes the nucleotide sequence shown in SEQ ID NO:238, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:298;

156)所述正义链包含如SEQ ID NO:268所示的核苷酸序列，所述反义链包含如SEQ ID NO:299所示的核苷酸序列；156) The sense strand includes the nucleotide sequence shown in SEQ ID NO:268, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:299;

157)所述正义链包含如SEQ ID NO:269所示的核苷酸序列，所述反义链包含如SEQ ID NO:299所示的核苷酸序列；157) The sense strand includes the nucleotide sequence shown in SEQ ID NO:269, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:299;

158)所述正义链包含如SEQ ID NO:268所示的核苷酸序列，所述反义链包含如SEQ ID NO:300所示的核苷酸序列；158) The sense strand includes the nucleotide sequence shown in SEQ ID NO:268, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:300;

159)所述正义链包含如SEQ ID NO:268所示的核苷酸序列，所述反义链包含如SEQ ID NO:301所示的核苷酸序列；159) The sense strand includes the nucleotide sequence shown in SEQ ID NO:268, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:301;

160)所述正义链包含如SEQ ID NO:268所示的核苷酸序列，所述反义链包含如SEQ ID NO:302所示的核苷酸序列；160) The sense strand includes the nucleotide sequence shown in SEQ ID NO:268, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:302;

161)所述正义链包含如SEQ ID NO:270所示的核苷酸序列，所述反义链包含如SEQ ID NO:303所示的核苷酸序列；161) The sense strand includes the nucleotide sequence shown in SEQ ID NO:270, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:303;

162)所述正义链包含如SEQ ID NO:271所示的核苷酸序列，所述反义链包含如SEQ ID NO:303所示的核苷酸序列；162) The sense strand includes the nucleotide sequence shown in SEQ ID NO:271, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:303;

163)所述正义链包含如SEQ ID NO:270所示的核苷酸序列，所述反义链包含如SEQ ID NO:304所示的核苷酸序列；163) The sense strand includes the nucleotide sequence shown in SEQ ID NO:270, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:304;

164)所述正义链包含如SEQ ID NO:270所示的核苷酸序列，所述反义链包含如SEQ ID NO:305所示的核苷酸序列； 164) The sense strand includes the nucleotide sequence shown in SEQ ID NO:270, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:305;

165)所述正义链包含如SEQ ID NO:270所示的核苷酸序列，所述反义链包含如SEQ ID NO:306所示的核苷酸序列；165) The sense strand includes the nucleotide sequence shown in SEQ ID NO:270, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:306;

166)所述正义链包含如SEQ ID NO:272所示的核苷酸序列，所述反义链包含如SEQ ID NO:307所示的核苷酸序列；166) The sense strand includes the nucleotide sequence shown in SEQ ID NO:272, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:307;

167)所述正义链包含如SEQ ID NO:273所示的核苷酸序列，所述反义链包含如SEQ ID NO:307所示的核苷酸序列；167) The sense strand includes the nucleotide sequence shown in SEQ ID NO:273, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:307;

168)所述正义链包含如SEQ ID NO:272所示的核苷酸序列，所述反义链包含如SEQ ID NO:308所示的核苷酸序列；168) The sense strand includes the nucleotide sequence shown in SEQ ID NO:272, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:308;

169)所述正义链包含如SEQ ID NO:272所示的核苷酸序列，所述反义链包含如SEQ ID NO:309所示的核苷酸序列；169) The sense strand includes the nucleotide sequence shown in SEQ ID NO:272, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:309;

170)所述正义链包含如SEQ ID NO:272所示的核苷酸序列，所述反义链包含如SEQ ID NO:310所示的核苷酸序列；170) The sense strand includes the nucleotide sequence shown in SEQ ID NO:272, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:310;

171)所述正义链包含如SEQ ID NO:274所示的核苷酸序列，所述反义链包含如SEQ ID NO:311所示的核苷酸序列；171) The sense strand includes the nucleotide sequence shown in SEQ ID NO:274, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:311;

172)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:311所示的核苷酸序列；172) 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:311;

173)所述正义链包含如SEQ ID NO:274所示的核苷酸序列，所述反义链包含如SEQ ID NO:312所示的核苷酸序列；173) The sense strand includes the nucleotide sequence shown in SEQ ID NO:274, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:312;

174)所述正义链包含如SEQ ID NO:274所示的核苷酸序列，所述反义链包含如SEQ ID NO:313所示的核苷酸序列；174) The sense strand includes the nucleotide sequence shown in SEQ ID NO:274, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:313;

175)所述正义链包含如SEQ ID NO:274所示的核苷酸序列，所述反义链包含如SEQ ID NO:314所示的核苷酸序列；175) The sense strand includes the nucleotide sequence shown in SEQ ID NO:274, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:314;

176)所述正义链包含如SEQ ID NO:275所示的核苷酸序列，所述反义链包含如SEQ ID NO:315所示的核苷酸序列；176) The sense strand includes the nucleotide sequence shown in SEQ ID NO:275, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:315;

177)所述正义链包含如SEQ ID NO:276所示的核苷酸序列，所述反义链包含如SEQ ID NO:316所示的核苷酸序列；177) The sense strand includes the nucleotide sequence shown in SEQ ID NO:276, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:316;

178)所述正义链包含如SEQ ID NO:277所示的核苷酸序列，所述反义链包含如SEQ ID NO:317所示的核苷酸序列；178) The sense strand includes the nucleotide sequence shown in SEQ ID NO:277, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:317;

179)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:318所示的核苷酸序列；179) 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:318;

180)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:319所示的核苷酸序列；180) 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:319;

181)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:320所示的核苷酸序列；181) 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:320;

182)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:321所示的核苷酸序列；182) 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:321;

183)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:322所示的核苷酸序。183) 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:322.

[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C3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 C3 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 modification shown in Table 2 with a conjugating group;

优选地，所述双链核糖核酸缀合物中，所述双链核糖核酸的正义链与反义链选自如下组合：Preferably, in the double-stranded ribonucleic acid conjugate, the sense strand and antisense strand of the double-stranded ribonucleic acid are selected from the following combinations:

3)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:133所示的核苷酸序列；3) 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:37 The nucleotide sequence shown in NO:133;

6)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:136所示的核苷酸序列；6) 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:136;

25)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:155所示的核苷酸序列； 25) 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:155;

29)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:159所示的核苷酸序列；29) 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:159;

48)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:178所示的核苷酸序列；48) 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:82 The nucleotide sequence shown in NO:178;

51)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:181所示的核苷酸序列；51) 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:181;

70)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:200所示的核苷酸序列； 70) 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:200;

74)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:204所示的核苷酸序列；74) 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:204;

93)所述正义链包含如SEQ ID NO:127所示的核苷酸序列，所述反义链包含如SEQ ID NO:223所示的核苷酸序列；93) The sense strand comprises the nucleotide sequence shown in SEQ ID NO:127, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:127 The nucleotide sequence shown in NO:223;

118)所述正义链包含如SEQ ID NO:328所示的核苷酸序列，所述反义链包含如SEQ ID NO:346所示的核苷酸序列；118) The sense strand includes the nucleotide sequence shown in SEQ ID NO:328, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:346;

135)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:363所示的核苷酸序列；135) 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:363;

183)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:322所示的核苷酸序列；183) 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:322;

任选地，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:257所示的序列，所述反义链包含如SEQ ID NO:260所示的核苷酸序列；Optionally, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:257, and the antisense strand includes the sequence shown in SEQ ID NO:260 nucleotide sequence;

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:258所示的序列，所述反义链包含如SEQ ID NO:261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 sequence shown in SEQ ID NO:258, and the antisense strand includes the core shown in SEQ ID NO:261 nucleotide sequence;

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:259所示的序列，所述反义链包含如SEQ ID NO:262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 sequence shown in SEQ ID NO:259, and the antisense strand includes the core shown in SEQ ID NO:262 nucleotide sequence;

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 所示的序列，所述反义链包含如SEQ ID NO:283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: 257 The sequence shown, the antisense strand includes the nucleotide sequence shown in SEQ ID NO:283;

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所示的序列，所述反义链包含如SEQ ID NO:284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 sequence shown in SEQ ID NO:257, and the antisense strand includes the core shown in SEQ ID NO:284 nucleotide sequence;

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所示的序列，所述反义链包含如SEQ ID NO:285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 sequence shown in SEQ ID NO:257, and the antisense strand includes the core shown in SEQ ID NO:285 nucleotide sequence;

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所示的序列，所述反义链包含如SEQ ID NO:286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 sequence shown in SEQ ID NO:257, and the antisense strand includes the core shown in SEQ ID NO:286 nucleotide sequence;

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:323所示的序列，所述反义链包含如SEQ ID NO:251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 sequence shown in SEQ ID NO:323, and the antisense strand includes the core shown in SEQ ID NO:251 nucleotide sequence;

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:323所示的序列，所述反义链包含如SEQ ID NO:287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 sequence shown in SEQ ID NO:323, and the antisense strand includes the core shown in SEQ ID NO:287 nucleotide sequence;

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:323所示的序列，所述反义链包含如SEQ ID NO:288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 sequence shown in SEQ ID NO: 323, and the antisense strand includes the core shown in SEQ ID NO: 288 nucleotide sequence;

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:323所示的序列，所述反义链包含如SEQ ID NO:289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 sequence shown in SEQ ID NO:323, and the antisense strand includes the core shown in SEQ ID NO:289 nucleotide sequence;

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:323所示的序列，所述反义链包含如SEQ ID NO:290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 sequence shown in SEQ ID NO:323, and the antisense strand includes the core shown in SEQ ID NO:290 nucleotide sequence;

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:258所示的序列，所述反义链包含如SEQ ID NO:291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 sequence shown in SEQ ID NO:258, and the antisense strand includes the core shown in SEQ ID NO:291 nucleotide sequence;

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:258所示的序列，所述反义链包含如SEQ ID NO:292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 sequence shown in SEQ ID NO:258, and the antisense strand includes the core shown in SEQ ID NO:292 nucleotide sequence;

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:258所示的序列，所述反义链包含如SEQ ID NO:293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 sequence shown in SEQ ID NO:258, and the antisense strand includes the core shown in SEQ ID NO:293 nucleotide sequence;

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:258所示的序列，所述反义链包含如SEQ ID NO:294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 sequence shown in SEQ ID NO:258, and the antisense strand includes the core shown in SEQ ID NO:294 nucleotide sequence;

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:259所示的序列，所述反义链包含如SEQ ID NO:295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 sequence shown in SEQ ID NO:259, and the antisense strand includes the core shown in SEQ ID NO:295 nucleotide sequence;

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:259所示的序列，所述反义链包含如SEQ ID NO:296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 sequence shown in SEQ ID NO:259, and the antisense strand includes the core shown in SEQ ID NO:296 nucleotide sequence;

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:259所示的序列，所述反义链包含如SEQ ID NO:297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 sequence shown in SEQ ID NO:259, and the antisense strand includes the core shown in SEQ ID NO:297 nucleotide sequence;

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:259所示的序列，所述反义链包含如SEQ ID NO:298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 sequence shown in SEQ ID NO:259, and the antisense strand includes the core shown in SEQ ID NO:298 nucleotide sequence;

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:324所示的序列，所述反义链包含如SEQ ID NO:299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 sequence shown in SEQ ID NO:324, and the antisense strand includes the core shown in SEQ ID NO:299 nucleotide sequence;

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:324所示的序列，所述反义链包含如SEQ ID NO:300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 sequence shown in SEQ ID NO:324, and the antisense strand includes the core shown in SEQ ID NO:300 nucleotide sequence;

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:324所示的序列，所述反义链包含如SEQ ID NO:301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 sequence shown in SEQ ID NO:324, and the antisense strand includes the core shown in SEQ ID NO:301 nucleotide sequence;

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:324所示的序列，所述反义链包含如SEQ ID NO:302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 sequence shown in SEQ ID NO:324, and the antisense strand includes the core shown in SEQ ID NO:302 nucleotide sequence;

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:325所示的序列，所述反义链包含如SEQ ID NO:303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 sequence shown in SEQ ID NO:325, and the antisense strand includes the core shown in SEQ ID NO:303 nucleotide sequence;

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:325所示的序列，所述反义链包含如SEQ ID NO:304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 sequence shown in SEQ ID NO:325, and the antisense strand includes the core shown in SEQ ID NO:304. nucleotide sequence;

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:325所示的序列，所述反义链包含如SEQ ID NO:305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 sequence shown in SEQ ID NO:325, and the antisense strand includes the core shown in SEQ ID NO:305 nucleotide sequence;

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:325所示的序列，所述反义链包含如SEQ ID NO:306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 sequence shown in SEQ ID NO:325, and the antisense strand includes the core shown in SEQ ID NO:306 nucleotide sequence;

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:326所示的序列，所述反义链包含如SEQ ID NO:307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 sequence shown in SEQ ID NO:326, and the antisense strand includes the core shown in SEQ ID NO:307 nucleotide sequence;

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:326所示的序列，所述反义链包含如SEQ ID NO:308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 sequence shown in SEQ ID NO:326, and the antisense strand includes the core shown in SEQ ID NO:308 nucleotide sequence;

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:326所示的序列，所述反义链包含如SEQ ID NO:309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 sequence shown in SEQ ID NO:326, and the antisense strand includes the core shown in SEQ ID NO:309 nucleotide sequence;

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:326所示的序列，所述反义链包含如SEQ ID NO:310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 sequence shown in SEQ ID NO:326, and the antisense strand includes the core shown in SEQ ID NO:310 nucleotide sequence;

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:327所示的序列，所述反义链包含如SEQ ID NO:311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 sequence shown in SEQ ID NO:327, and the antisense strand includes the core shown in SEQ ID NO:311 nucleotide sequence;

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:327所示的序列，所述反义链包含如SEQ ID NO:312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 sequence shown in SEQ ID NO:327, and the antisense strand includes the core shown in SEQ ID NO:312 nucleotide sequence;

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:327所示的序列，所述反义链包含如SEQ ID NO:313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 sequence shown in SEQ ID NO:327, and the antisense strand includes the core shown in SEQ ID NO:313 nucleotide sequence;

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:327所示的序列，所述反义链包含如SEQ ID NO:314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 sequence shown in SEQ ID NO:327, and the antisense strand includes the core shown in SEQ ID NO:314. nucleotide sequence.

[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C3基因表达，或制备用于抑制C3基因表达的药物；(1) Inhibit C3 gene expression, or prepare drugs for inhibiting C3 gene expression;

(2)用于预防或治疗与C3基因异常表达相关的疾病，或制备用于预防或治疗与C3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 C3 gene, or preparing drugs for preventing or treating diseases related to abnormal expression of C3 gene;

(3)用于治疗患有将受益于补体C3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补体C3基因表达降低的疾病的受试者的药物。(3) For use in treating a subject suffering from a disease that would benefit from a reduction in complement C3 gene expression, or to prepare a medicament for treating a subject suffering from a disease that would benefit from a reduction in complement C3 gene expression.

[40].根据[39]所述的用途，其中，所述与C3基因异常表达相关的疾病选自如下疾病组成的组：[40]. The use according to [39], wherein the disease associated with abnormal expression of the C3 gene is selected from the group consisting of the following diseases:

阵发性睡眠性血红蛋白尿症、非典型溶血性尿毒症综合征、类风湿性关节炎、慢性肾病、缺血再灌注损伤、神经退行性疾病、哮喘、系统性红斑狼疮、肾小球肾炎、银屑病、皮肌炎大疱性类天疱疮、志贺毒素大肠杆菌相关的溶血性尿毒症综合征、重症肌无力、视神经脊髓炎、致密物沉积病、C3神经疾病、年龄相关性黄斑变性、冷凝集素病、抗中性粒细胞胞浆抗体相关性血管炎、体液和血管移植排斥、移植物功能障碍、心肌梗塞、移植物的敏化受体和脓毒症。Paroxysmal nocturnal hemoglobinuria, atypical hemolytic uremic syndrome, rheumatoid arthritis, chronic kidney disease, ischemia-reperfusion injury, neurodegenerative diseases, asthma, systemic lupus erythematosus, glomerulonephritis, Psoriasis, dermatomyositis bullous pemphigoid, Shiga toxin E. coli-related hemolytic uremic syndrome, myasthenia gravis, neuromyelitis optica, dense deposit disease, C3 neurological disease, age-related macular degeneration, cold agglutinin disease, antineutrophil cytoplasmic antibody-associated vasculitis, humoral and vascular graft rejection, graft dysfunction, myocardial infarction, sensitized recipients of grafts, and sepsis.

[41].一种用于抑制细胞内C3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 intracellular C3 gene expression, 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C3基因异常表达，更具体地为C3基因异常高表达；Abnormal expression of C3 gene in the body, more specifically abnormal high expression of C3 gene;

患有与C3基因异常表达相关的疾病；Suffering from diseases related to abnormal expression of C3 gene;

患有将受益于C3基因表达降低的疾病。People with diseases that would benefit from reduced C3 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)，切割补体C3基因转录的mRNA，高效、特异地抑制补体C3基因的表达，用于治疗补体系统的不适当的激活介导的疾病。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 complement C3 gene, and efficiently and specifically inhibit the expression of the complement C3 gene. For the treatment of diseases mediated by inappropriate activation of the complement system.

进一步的，本公开中双链核糖核酸为siRNA，siRNA靶向结合并降解C3基因的转录产物mRNA，发挥RNA干扰的作用，抑制C3基因的蛋白表达，是一种抑制率高且特异性好的C3补体抑制剂。Furthermore, in the present disclosure, the double-stranded ribonucleic acid is siRNA. siRNA targets and degrades the transcript product of the C3 gene, mRNA, and exerts the effect of RNA interference to inhibit the protein expression of the C3 gene. It is a method with high inhibition rate and good specificity. C3 complement inhibitor.

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C3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 the C3 gene in the liver.

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“C3”、“补体C3”、“C3补体”“complement C3(C3)”，是指熟知的基因和多肽，也在本领域已知为：ARMD9、C3a过敏毒素、ASP、补体成分C3a、C3a、补体成分C3b、C3b、前体原-C3、酰化刺激蛋白切割产物、CPAMD1、补体 C3、C3以及含蛋白质1的PZP样α-2-巨球蛋白结构域、补体C3以及AHUS5。C3基因、C3mRNA序列的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 "C3", "complement C3", "complement C3", "complement C3(C3)", as used in the context of this disclosure, refer to well-known genes and polypeptides also known in the art as: ARMD9, C3a anaphylatoxin, ASP, complement component C3a, C3a, complement component C3b, C3b, prepro-C3, acylation stimulating protein cleavage product, CPAMD1, complement C3, C3 and PZP-like α-2-macroglobulin domain containing protein 1, complement C3 and AHUS5. The C3 gene and C3 mRNA sequences are easily obtained using, for example, the following: GenBank (GenBank), database (UniProt), Online Mendelian Inheritance in Man (OMIM), etc.

术语“C3基因”，可以是野生型C3基因，或存在序列变异的C3基因突变体。在C3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 "C3 gene" can be a wild-type C3 gene or a C3 gene mutant with sequence variation. Many sequence variations in the C3 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 amino acid polymers that have been modified (eg, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation, such as conjugation with a labeling component).

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、22、23、24、25、27、28、34、35、38、50、57、58、61、106、340、385、441、548、584或633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 sequence is 19, 20, 22, 23, 24, 25, 27, 28, 34, 35, 38, 50, 57, 58, 61, 106, 340, 385, 441, 548, 584 or 633 A sequence of nucleotides consisting of consecutively linked 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C3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C3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 C3 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 C3 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C3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 a "normal" relative to a target gene, such as the C3 gene. "sense" and "antisense" orientations. 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 compounds in which the position of the base in the ribose ring is changed in the nucleotide, for example, the base is not attached to the 1'-position of the ribose ring, but is A compound formed by linking 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C3mRNA)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 RNA that is substantially complementary to a target sequence (eg, derived from human C3 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 complementary" 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. Correspondingly, "mismatch" in this field 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C3基因的表达”包括抑制任何C3基因(如例如小鼠C3基因、大鼠C3基因、猴C3基因、或人类C3基因)以及C3基因的变体(例如天然存在的变体)或突变体的表达。因此，该C3基因可以是野生型C3基因、突变C3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C3基因。The term "inhibiting the expression of a C3 gene" includes inhibiting any C3 gene (such as, for example, a mouse C3 gene, a rat C3 gene, a monkey C3 gene, or a human C3 gene) as well as variants (eg, naturally occurring variants) of the C3 gene, or Expression of mutants. Thus, the C3 gene may be a wild-type C3 gene, a mutant C3 gene, or in the case of a genetically manipulated cell, cell population or organism, a transgenic C3 gene.

“抑制C3基因表达”包括任何水平的C3基因的抑制，例如至少部分抑制C3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 C3 gene expression" includes any level of inhibition of the C3 gene, such as at least partial inhibition of the expression of the C3 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，“C_1-6烷基”中的“C_1-6”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, the "C _1-6 " in "C _1-6 alkyl" refers to a linear, branched or cyclic arrangement containing 1, 2, 3, 4, 5 or 6 carbon atoms. group.

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 dosage regimen selected; and the use of any concomitant therapy.

术语“与C3基因异常表达相关的疾病”是由补体激活引起或与补体激活相关联的疾病或障碍。术语“与C3基因异常表达相关的疾病”包括将从减少C3(即“C3-相关性疾病”)表达受益的疾病、障碍或病症。此类疾病典型地与炎症和/或免疫系统激活，例如膜攻击复合体介导的溶解、过敏反应和/或溶血相关联。补体成分相关性疾病的非限制性实例包括阵发性睡眠性血红蛋白尿症(PNH)、非典型溶血性尿毒症综合征(aHUS)、哮喘、类风湿性关节炎(RA)；抗磷脂抗体综合征；狼疮性肾炎；缺血再灌注损伤；典型的或侵染性的溶血性尿毒症综合征(tHUS)；致密物沉积病(DDD)；视神经脊髓炎(NMO)；多灶运动神经病变(MMN)；多发性硬化症(MS)；黄斑变性(例如，年龄相关性黄斑变性(AMD))；溶血，肝酶升高以及低血小板(HELLP)综合征；血栓性血小板减少性紫癜(TTP)；自发流产；寡免疫性血管炎；大疱性表皮松解症；习惯性流产；先兆子痫、创伤性脑损伤、重症肌无力、冷凝集素病、皮肌炎大疱性类天疱疮、志贺毒素大肠杆菌相关的溶血性尿毒症综合征、C3神经病、抗嗜中性粒细胞细胞质抗体相关联的血管炎、体液和血管移植排斥、移植物功能异常、心肌梗塞、异源移植、脓毒症、冠状动脉病、皮肌炎、格雷夫斯病、动脉粥样硬化、阿耳茨海默氏病、全身性炎症反应脓毒症、脓毒性休克、脊髓损伤、肾小球性肾炎、桥本氏甲状腺炎、I型糖尿病、牛皮癣、天疱疮、自身免疫性溶血性贫血(AIHA)、ITP、肺出血肾炎综合征、德戈斯病、抗磷脂综合征(APS)、灾难性APS(CAPS)、心血管障碍、心肌炎、脑血管障碍、外周(例如，肌肉骨骼)血管障碍、肾血管障碍、肠系膜/肠血管障碍、血管炎、亨诺赫-舍恩莱因紫癜肾炎、全身性红斑狼疮相关联的血管炎、与类风湿性关节炎相关联的血管炎、免疫复合物血管炎、高安氏病、扩张性心肌病、糖尿病性血管病、川畸氏病(动脉炎)、静脉气体栓子(VGE)以及支架放置、旋转式动脉粥样斑块去除术、和经皮穿刺冠状动脉成形术(PTCA)后的再狭窄(参见，例如，郝勒斯(Holers)(2008)免疫学评论(ImmunologicalReviews)223:300-316；郝勒斯(Holers)和帕德兰(Thurman)(2004)分子免疫学(MolecularImmunology)41:147-152；美国专利公开号20070172483)。The term "disease associated with abnormal expression of the C3 gene" is a disease or disorder caused by or associated with complement activation. The term "disease associated with abnormal expression of the C3 gene" includes diseases, disorders or conditions that would benefit from reduced expression of C3 (i.e., "C3-related diseases"). Such diseases are typically associated with inflammation and/or immune system activation, such as membrane attack complex-mediated lysis, allergic reactions, and/or hemolysis. Non-limiting examples of complement component-related diseases include paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), asthma, rheumatoid arthritis (RA); antiphospholipid antibody syndrome Symptoms; lupus nephritis; ischemia-reperfusion injury; classic or invasive hemolytic uremic syndrome (tHUS); dense deposit disease (DDD); neuromyelitis optica (NMO); multifocal motor neuropathy ( MMN); multiple sclerosis (MS); macular degeneration (e.g., age-related macular degeneration (AMD)); hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome; thrombotic thrombocytopenic purpura (TTP) ; Spontaneous abortion; Pauciimmune vasculitis; Epidermolysis bullosa; Recurrent abortion; Preeclampsia, traumatic brain injury, myasthenia gravis, cold agglutinin disease, dermatomyositis bullous pemphigoid , Shiga toxin E. coli related hemolytic uremia syndrome, C3 neuropathy, antineutrophil cytoplasmic antibody-associated vasculitis, humoral and vascular graft rejection, graft dysfunction, myocardial infarction, allogeneic transplantation, sepsis, coronary artery disease, dermatomyositis , Graves' disease, atherosclerosis, Alzheimer's disease, systemic inflammatory response, sepsis, septic shock, spinal cord injury, glomerulonephritis, Hashimoto's thyroiditis, type I diabetes, Psoriasis, pemphigus, autoimmune hemolytic anemia (AIHA), ITP, pulmonary hemorrhage nephritis syndrome, Degos disease, antiphospholipid syndrome (APS), catastrophic APS (CAPS), cardiovascular disorders, myocarditis, Cerebrovascular disorders, peripheral (e.g., musculoskeletal) vascular disorders, renovascular disorders, mesenteric/intestinal vascular disorders, vasculitis, Henoch-Schoenlein purpura nephritis, vasculitis associated with systemic lupus erythematosus, and similar Rheumatoid arthritis-associated vasculitis, immune complex vasculitis, Takayasu's disease, dilated cardiomyopathy, diabetic vasculopathy, Kawasaki disease (arteritis), venous gas emboli (VGE), and stent placement, Restenosis after rotational atherectomy, and percutaneous coronary angioplasty (PTCA) (see, eg, Holers (2008) Immunological Reviews 223:300-316 ; Holers and Thurman (2004) Molecular Immunology 41:147-152; U.S. Patent Publication No. 20070172483).

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)，用于抑制补体C3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，C3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，C3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 the complement C3 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 process of the target gene (i.e., the C3 gene) and is used to guide the transcript of the target mRNA (i.e., the C3 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，抑制补体C3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 complement C3 gene.

根据来源于人C3mRNA(NM_000064.4)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:8-34、SEQ ID NO:364任一项所示的核苷酸序列。Based on the target sequence derived from human C3 mRNA (NM_000064.4), 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 shown in any one of SEQ ID NO:8-34 and SEQ ID NO:364.

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:8～9、 SEQ ID NO:364所示的核苷酸序列。In some specific embodiments, the nucleotide sequence shown in SEQ ID NO:1 includes SEQ ID NO:8～9, The nucleotide sequence shown in SEQ ID NO:364.

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:10～13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:10-13.

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:14～16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:14-16.

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:17～20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:17～20.

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:21～24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:21～24.

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:25～30所示的核苷酸序列。In some specific embodiments, the nucleotide sequence shown in SEQ ID NO: 6 includes the nucleotide sequence shown in SEQ ID NO: 25-30.

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:31～34所示的核苷酸序列。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 specific embodiments, the nucleotide sequence shown in SEQ ID NO:7 includes the nucleotide sequence shown in SEQ ID NO:31～34. 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。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.

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；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 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, etc.; in some preferred embodiments In , the length ratio of sense strand/antisense strand is 19/21 or 21/23.

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:34、SEQ ID NO:364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:34、SEQ ID NO:364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:34、SEQ ID NO:364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。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 ~ SEQ ID NO:34, SEQ ID NO:364 and siRNA that can be complementary to these target sequences, for each siRNA's antisense strand, includes its complementary target The sequence (for example, any one of SEQ ID NO: 8 ~ SEQ ID NO: 34, SEQ ID NO: 364) is basically complementary, that is, the nucleotide sequence of the antisense strand of each siRNA can There is a base mismatch with the corresponding target sequence; for each sense strand of siRNA, it includes the target sequence that is complementary to it (for example, SEQ ID NO:8 ~ SEQ ID NO:34, SEQ ID NO:364 (any of) are substantially complementary, that is, the nucleotide sequence of the sense strand of each siRNA may have a base mismatch 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.

示例性的，双链核糖核酸为表1中示出的siRNA1。Exemplarily, the double-stranded ribonucleic acid is siRNA1 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: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.

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:9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:9, and the nucleotide sequence of the antisense strand is SEQ ID NO:9 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:9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:9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。示例性的，双链核糖核酸为表1中示出的siRNA2～siRNA10、siRNA118～siRNA128、siRNA130～siRNA132、siRNA134、siRNA135中的任一siRNA。 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:9, and the nucleotide sequence of the antisense strand is SEQ ID NO:9 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. Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA2 to siRNA10, siRNA118 to siRNA128, siRNA130 to siRNA132, siRNA134, and siRNA135 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:10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:10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:10, and the nucleotide sequence of the antisense strand is SEQ ID NO:10 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: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siRNA11～siRNA13中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any one of siRNA11 to siRNA13 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.

示例性的，双链核糖核酸为表1中示出的siRNA14。Exemplarily, the double-stranded ribonucleic acid is siRNA14 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:12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:12, and the nucleotide sequence of the antisense strand is SEQ ID NO:12 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:12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:12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:12, and the nucleotide sequence of the antisense strand is SEQ ID NO:12 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: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中示出的siRNA15～siRNA21中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA15 to siRNA21 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: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中示出的siRNA22～siRNA23中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any one of siRNA22 to siRNA23 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: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中示出的siRNA24～siRNA26中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA24 to siRNA26 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: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siRNA27～siRNA29中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA27 to siRNA29 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siRNA30。Exemplarily, the double-stranded ribonucleic acid is 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: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中示出的siRNA31～siRNA32中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA31 to siRNA32 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: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 sequence A consisting of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:18, and the nucleotide sequence of the antisense strand is SEQ ID NO:18 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:18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:18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:18, and the nucleotide sequence of the antisense strand is SEQ ID NO:18 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:18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:18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:18, and the nucleotide sequence of the antisense strand is SEQ ID NO:18 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siRNA33～siRNA41中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any one of siRNA33 to siRNA41 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.

示例性的，双链核糖核酸为表1中示出的siRNA42。Exemplarily, the double-stranded ribonucleic acid is siRNA42 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siRNA43～siRNA44中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any one of siRNA43 to siRNA44 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中示出的siRNA45～siRNA46中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA45 to siRNA46 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: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SEQ ID NO:22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: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。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, and the nucleotide sequence of the antisense strand is SEQ ID NO:22 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.

示例性的，双链核糖核酸为表1中示出的siRNA47～siRNA50中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA47 to siRNA50 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: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 the 19 consecutive nucleotides in the sequence shown in SEQ ID NO:23. The nucleotide sequence of the antisense strand is sequence A, which is composed of continuation nucleotides. The nucleotide sequence of the antisense strand is sequence B, which is the reverse complement of the sequence B composed of 19 consecutive nucleotides in the sequence shown in SEQ ID NO:23. 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siRNA51～siRNA53中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA51 to siRNA53 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.

示例性的，双链核糖核酸为表1中示出的siRNA54～siRNA55中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA54 to siRNA55 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: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.

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:25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:25, and the nucleotide sequence of the antisense strand is SEQ ID NO:25 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.

示例性的，双链核糖核酸为表1中示出的siRNA56～siRNA58中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA56 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:26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:26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:26, and the nucleotide sequence of the antisense strand is SEQ ID NO:26 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～siRNA60中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA59 to siRNA60 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:27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:27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:27, and the nucleotide sequence of the antisense strand is SEQ ID NO:27 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siRNA61。Exemplarily, the double-stranded ribonucleic acid is siRNA61 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:28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:28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:28, and the nucleotide sequence of the antisense strand is SEQ ID NO:28 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siRNA62～siRNA63中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA62 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:29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:29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:29, and the nucleotide sequence of the antisense strand is SEQ ID NO:29 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siRNA64～siRNA65中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA64 to siRNA65 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:30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:30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:30, and the nucleotide sequence of the antisense strand is SEQ ID NO:30 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:30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:30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:30, and the nucleotide sequence of the antisense strand is SEQ ID NO:30 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:30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:30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:30, and the nucleotide sequence of the antisense strand is SEQ ID NO:30 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siRNA66～siRNA86中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA66 to siRNA86 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:31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:31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:31, and the nucleotide sequence of the antisense strand is SEQ ID NO:31 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siRNA87。Exemplarily, the double-stranded ribonucleic acid is siRNA87 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:32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:32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:32, and the nucleotide sequence of the antisense strand is SEQ ID NO:32 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siRNA88～siRNA89中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any siRNA among siRNA88 to siRNA89 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:33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:33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:33, and the nucleotide sequence of the antisense strand is SEQ ID NO:33 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siRNA90～siRNA94中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any one of siRNA90 to siRNA94 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:34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:34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:34, and the nucleotide sequence of the antisense strand is SEQ ID NO:34 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siRNA95。Exemplarily, the double-stranded ribonucleic acid is siRNA95 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:364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:364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:364, and the nucleotide sequence of the antisense strand is SEQ ID NO:364 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:364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:364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:364, and the nucleotide sequence of the antisense strand is SEQ ID NO:364 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siRNA129、siRNA133中的任一siRNA。Illustratively, the double-stranded ribonucleic acid is any one of siRNA129 and siRNA133 shown in Table 1.

在一些具体的实施方式中，所述正义链包含如SEQ ID NO:35-129、SEQ ID NO:328～345任一项所示的核苷酸序列，所述反义链包含如SEQ ID NO:131-225、SEQ ID NO:346～363任一项所示的核苷酸序列。In some specific embodiments, the sense strand includes the nucleotide sequence shown in any one of SEQ ID NO:35-129 and SEQ ID NO:328-345, and the antisense strand includes the nucleotide sequence shown in any one of SEQ ID NO:35-129 and SEQ ID NO:328-345. :131-225, the nucleotide sequence shown in any one of SEQ ID NO:346~363.

在一些具体的实施方案中，双链核糖核酸选自如表1中所示的任一siRNA。本公开提供的siRNA，其与靶mRNA(C3mRNA)结合的特异性高，具有较好的靶mRNA的沉默活性，可以显著抑制补体C3基因表达，用于治疗补体系统的不适当的激活介导的疾病。In some specific embodiments, the double-stranded ribonucleic acid is selected from any of the siRNAs shown in Table 1. The siRNA provided by the present disclosure has high binding specificity to the target mRNA (C3 mRNA), has good silencing activity of the target mRNA, can significantly inhibit complement C3 gene expression, and is used to treat inappropriate activation of the complement system. 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.

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 core. glycosides. 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. 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C3 mRNA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 C3 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.Dr ug 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C3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. Dr ug 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 C3 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、m^2,2G、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。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, 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. 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:

(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 comprising 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, the 3' end of sequence A has When there are 1-2 protruding nucleotides extending out of the double-stranded region, the protruding nucleotide at the 3' end of sequence A is excluded and the nucleotide sequence of the sense strand is used. 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 nucleotide sequence of the sense strand of the double-stranded ribonucleic acid modification 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 and 3rd nucleotides 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',

(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 ):

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, 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 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',

(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' ,

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:227-241、263-265、266-282任一项所示的核苷酸序列，并且所述反义链包含如SEQ ID NO:242-256、260-262、283-322任一项所示的核苷酸序列。In some optional embodiments, the sense strand comprises a nucleotide sequence as shown in any one of SEQ ID NOs: 227-241, 263-265, 266-282, and the antisense strand comprises a nucleotide sequence as shown in any one of SEQ ID NOs: 227-241, 263-265, 266-282 ID NO: 242-256, 260-262, 283-322 any one of the nucleotide sequences.

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。Exemplarily, 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 chain 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 represented by sequence A', and the nucleotide sequence of the antisense strand is the sequence represented 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 ):

(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'，

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 2 and connect it to the 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：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-acetyl galactosamine linked through nucleosides elicit robust gene silencing in vivo in hepatocytes.A CS Chemical biology,2015,10(5):1181-7。Muthiah Manoharan et.al.siRNA conjugates carrying sequentially assembled trivalent N-acetyl galactosamine linked through nucleosides elicit robust gene silencing in vivo in hepatocytes.A CS 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C3基因的高表达。The conjugation group shown in Formula I is GalNAc. GalNAc has liver-targeting properties and can deliver siRNA molecules to liver tissue with high specificity and specifically inhibit the high expression of the C3 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).

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 double-stranded ribonucleic acid, double-stranded ribonucleic acid modifications or double-stranded ribonucleic acid conjugates at least one of the following uses:

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，C3基因异常表达，导致补体系统的不适当激活，引发如下一种或多种C3基因异常表达相关疾病：阵发性睡眠性血红蛋白尿症、非典型溶血性尿毒症综合征、类风湿性关节炎、慢性肾病、缺血再灌注损伤、神经退行性疾病、哮喘、系统性红斑狼疮、肾小球肾炎、银屑病、皮肌炎大疱性类天疱疮、志贺毒素大肠杆菌相关的溶血性尿毒症综合征、重症肌无力、视神经脊髓炎、致密物沉积病、C3神经疾病、年龄相关性黄斑变性、冷凝集素病、抗中性粒细胞胞浆抗体相关性血管炎、体液和血管移植排斥、移植物功能障碍、心肌梗塞、移植物的敏化受体和脓毒症。In the present disclosure, abnormal expression of the C3 gene leads to inappropriate activation of the complement system, causing one or more of the following diseases related to abnormal expression of the C3 gene: paroxysmal nocturnal hemoglobinuria, atypical hemolytic uremic syndrome, Rheumatoid arthritis, chronic kidney disease, ischemia-reperfusion injury, neurodegenerative diseases, asthma, systemic lupus erythematosus, glomerulonephritis, psoriasis, dermatomyositis bullous pemphigoid, Shiga toxin Escherichia coli-related hemolytic uremic syndrome, myasthenia gravis, neuromyelitis optica, dense deposit disease, C3 neurological disease, age-related macular degeneration, cold agglutinin disease, antineutrophil cytoplasmic antibody-related vascular disease inflammation, humoral and vascular graft rejection, graft dysfunction, myocardial infarction, sensitized graft recipients, and sepsis.

siRNA分子致使C3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C3基因异常表达相关疾病的治疗。The siRNA molecule causes the expression of the C3 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 C3 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C3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 C3 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C3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 C3 gene in cells is to introduce siRNA molecules (including unmodified siRNA, siRNA modifications, and 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C3基因异常表达，更具体地为C3基因异常高表达；(1) Abnormal expression of C3 gene in the body, more specifically abnormal high expression of C3 gene;

(2)患有与C3基因异常表达相关的疾病；(2) Suffering from diseases related to abnormal expression of C3 gene;

(3)患有将受益于C3基因表达降低的疾病。如罹患或倾向于患上与C3基因异常表达相关的疾病的人。(3) Suffering from diseases that would benefit from reduced C3 gene expression. Such as people who suffer from or are prone to suffer from diseases related to abnormal expression of C3 gene.

表1 siRNA序列信息

Table 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 as the letter s Two adjacent nucleotides are connected by a phosphorothioate group; P1: indicates that the nucleotide adjacent to the right of P1 is a 5'-phosphate nucleotide, (GNA) indicates that the adjacent nucleotide on the right A 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 as the letter s The two adjacent nucleotides on the left and right are connected by a phosphorothioate group; P1: indicates that the nucleotide adjacent to the right of P1 is a 5'-phosphate nucleotide, (GNA) indicates the phase on the right The adjacent ribonucleotide is a ribonucleotide modified with GNA; 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、试剂及仪器耗材如表4、表5所示：The siRNA, siRNA modifications and siRNA conjugates involved in the following examples were synthesized by Tianlin Biotechnology (Shanghai) Co., Ltd. The cells, reagents and instrument consumables used in the examples are as shown in Table 4 and Table 5:

表4
Table 4

表5

table 5

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C3基因mRNA序列，选择不同位点设计多对C3siRNAs，设计的所有单个siRNA均能靶向靶基因的所有转录本(如表6)，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 C3 gene mRNA sequence, multiple pairs of C3 siRNAs were designed at different sites. All single siRNAs designed can target all transcripts of the target gene (as shown in Table 6). These pairs of siRNAs were compared with other pairs by sequence similarity software. All non-target gene sequences have minimal homology.

表6
Table 6

用于设计siRNA的靶标序列如下所示，靶标序列来源于C3的基因mRNA序列(参见NM_000064.4)。The target sequence used to design siRNA is shown below. The target sequence is derived from the gene mRNA sequence of C3 (see NM_000064.4).

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II-3:
Target sequence II-3:

靶标序列II-4:
Target sequence II-4:

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III-3：
Target sequence III-3:

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IV-3：
Target sequence IV-3:

靶标序列IV-4：
Target sequence IV-4:

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V-4：
Target sequence V-4:

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VI-4：
Target sequence VI-4:

靶标序列VI-5：
Target sequence VI-5:

靶标序列VI-6：
Target sequence VI-6:

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:

靶标序列VII-4：
Target sequence VII-4:

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 vulcanization reagents. It 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C3基因表达Example 3: siRNA and siRNA modifications inhibit C3 gene expression

3.1实验材料： 3.1 Experimental materials:

Hep3B细胞，购自ATCC，货号HB-8064；Hep3B cells were purchased from ATCC, catalog number HB-8064;

EMEM培养基，购自ATCC，货号30-2003；EMEM medium, purchased from ATCC, product number 30-2003;

Lipofectamine RNAiMax转染试剂，购自Invitrogen，货号13778-150；Lipofectamine RNAiMax 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;

SiNTC(No target control)：与靶标不相关的RNA。SiNTC (No target control): RNA not related to the target.

3.2实验方法：3.2 Experimental methods:

3.2.1将Hep3B细胞铺于96孔板的新鲜EMEM培养基中培养24小时。将所培养的细胞用无PS(青霉素链霉素混合液)的EMEM培养基重悬，制成密度为5.55×10⁴/ml的细胞悬液，铺到96孔板中，每孔加90μL细胞悬液，即5000个细胞/孔。3.2.1 Plate Hep3B cells in fresh EMEM medium in a 96-well plate and culture them for 24 hours. Resuspend the cultured cells in EMEM medium without PS (penicillin and streptomycin mixture) to make a cell suspension with a density of 5.55×10 ⁴ /ml, spread it into a 96-well plate, and add 90 μL of cells to each well. suspension, i.e. 5000 cells/well.

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 siRNA母液。3.2.2 Centrifuge the dry powder of the 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 with ultrapure distilled water (ULtraPure Distilled Water) to prepare 100 μM siRNA. mother liquor.

3.2.3配制0.1nM的siRNA转染稀释液3.2.3 Prepare 0.1nM siRNA transfection dilution solution

(1)0.1μM siRNA贮备液制备：(1) Preparation of 0.1μM siRNA stock solution:

a)、取上述步骤3.2.2中制得的100μM的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 siRNA dilution with a final concentration of 1 μM;

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；c). Take 2 μl of the 1 μM siRNA dilution prepared in step b) and add 18 μl of ultrapure distilled water to obtain a siRNA stock solution with a final concentration of 0.1 μM;

(2)取步骤c)中制得的0.1μM的siRNA贮备液2μl，加入98μl Opti-medium，得到2nM的siRNA稀释液；(2) Take 2 μl of the 0.1 μM siRNA stock solution prepared in step c), and add 98 μl Opti-medium to obtain a 2 nM siRNA dilution;

(3)将转染试剂3ul，加入97ul Opti-medium，得到转染试剂稀释液；将Lipofectamine RNAiMax转染试剂稀释液与siRNA稀释液以1:1体积比混合，静置5分钟，将10μl的转染混合物加入到96孔板中转染所培养的Hep3B细胞(终体积100μl，该体系中siRNA的浓度为0.1nM)。(3)will 3ul of transfection reagent, add 97ul Opti-medium, get Transfection reagent diluent; mix Lipofectamine RNAiMax transfection reagent diluent and siRNA diluent at a volume ratio of 1:1, let stand for 5 minutes, add 10 μl of the transfection mixture to the 96-well plate to transfect the cultured Hep3B cells (The final volume is 100 μl, and the concentration of siRNA in this system is 0.1 nM).

3.2.4转染后培养48小时，每个浓度设置3个重复。3.2.4 Cultivate for 48 hours after transfection, and set 3 replicates for each concentration.

3.2.5利用TaqMan^TM Fast Advanced Cells-to-CT^TM试剂盒(Invitrogen#A35377)提取RNA：3.2.5 Extract RNA using TaqMan ^TM Fast Advanced Cells-to-CT ^TM Kit (Invitrogen#A35377):

a)吸掉每孔中的旧培养基，加入100μl PBS缓冲液洗涤两次；a) Aspirate off the old culture medium in each well, add 100μl PBS buffer and wash twice;

b)吸掉PBS缓冲液，每孔中加入50μl裂解液；b) Aspirate off the PBS buffer and add 50 μl of lysis solution to each well;

c)室温下以500rpm震荡孵育5分钟；c) Incubate at room temperature with shaking at 500rpm for 5 minutes;

d)每孔加入5μl终止液，室温下以500rpm震荡2分钟。d) Add 5 μl of stop solution to each well and shake at 500 rpm for 2 minutes at room temperature.

3.2.6对RNA进行逆转录至cDNA，按照以下步骤进行：3.2.6 To reverse transcribe RNA into cDNA, follow the following steps:

a)如下表所述制备逆转录反应混合物并充分混匀，在整个操作过程中，所有试剂都放置在冰上；a) Prepare the reverse transcription reaction mixture as described in the table below and mix thoroughly. During the entire operation, all reagents are placed on ice;

表7
Table 7

b)如下所述进行逆转录程序b) Perform the reverse transcription procedure as follows

37℃，30min；95℃，5min；4℃，静置。37℃, 30min; 95℃, 5min; 4℃, let stand.

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进行实时PCR分析3.2.7 Perform real-time PCR analysis

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

50℃，2分钟；50℃, 2 minutes;

95℃，20秒；95℃, 20 seconds;

95℃，1秒；60℃ 20秒(40个循环)。95℃, 1 second; 60℃, 20 seconds (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：
ΔCt＝Ct(目的基因)–Ct(ACTB)
ΔΔCt＝ΔCt(检测样品组)-ΔCt(Mock组)b) Use the following formula to calculate the relative expression of genes:
ΔCt=Ct(target gene)–Ct(ACTB)
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：和实验组相比，未加入siRNA的组。Among them, the Mock group indicates: compared with the experimental group, the group without siRNA was added.

3.3沉默实验结果3.3 Silence experiment results

选取浓度0.1nM进行测试Select a concentration of 0.1nM 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C3基因优异的抑制效果。在0.1nM的浓度下，抑制率高达93.22％。As can be seen from Table 9, the siRNA and siRNA modifications provided by the present disclosure show excellent inhibitory effects on the C3 gene. At a concentration of 0.1nM, the inhibition rate was as high as 93.22%.

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)：2、0.5、0.125、0.0313、0.00781、 0.00195、0.000488、0.000122，再按照与3.2相似的方法进行IC₅₀测定。The following siRNA measurement concentration range settings to be tested (nM): 2, 0.5, 0.125, 0.0313, 0.00781, 0.00195, 0.000488, 0.000122, and then measure IC ₅₀ according to a method similar to 3.2.

结果分析：Result analysis:

b)使用以下公式计算基因的相对表达量：
ΔCt＝Ct(C3基因)–Ct(ACTB)
ΔΔCt＝ΔCt(检测样品组)–ΔCt(Mock组)；b) Use the following formula to calculate the relative expression of genes:
ΔCt=Ct(C3 gene)–Ct(ACTB)
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–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 sample group)/relative expression of mRNA in the Mock group × 100%;

其中，Mock组表示和实验组相比，未加入siRNA的组。Among them, the Mock group represents the group in which siRNA was not added compared with the experimental 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、siRNA修饰物具有优异的C3基因抑制活性，可以在低至0.01778nM的IC₅₀时依然具有良好的C3基因抑制活性。As can be seen from Table 10, the siRNA and siRNA modifications provided by the present disclosure have excellent C3 gene inhibitory activity, and can still have good C3 gene inhibitory activity at an IC ₅₀ as low as 0.01778nM.

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

Lipofectamine RNAiMax转染试剂，购自Invitrogen，货号：13778-150；Lipofectamine RNAiMax transfection reagent, purchased from Invitrogen, product number: 13778-150;

RNA提取试剂盒96Kit(12)，货号：QIAGEN-74182；RNA extraction kit 96Kit(12), item number: QIAGEN-74182;

逆转录试剂盒FastKing RT Kit(With gDNase)，货号：天根-KR116-02；Reverse transcription kit FastKing RT Kit (With gDNase), product number: Tiangen-KR116-02;

FastStart Universal Probe Mast(Roche-04914058001)；C3及GAPDH引物由药明康德提供。FastStart Universal Probe Mast (Roche-04914058001); C3 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Lipofectamine RNAiMax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RNA提取。根据说明书使用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 use Lipofectamine RNAiMax to transfer the siRNA conjugate into the cells, and seed them into a 96-well plate at a density of 54,000 cells per well. Add 100 μL of PPH medium 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 RNA extraction. Use according to 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RNA提取。根据说明书使用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 freeze The stored 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 RNA extraction. Use according to instructions 96Kit extracted total RNA.

通过逆转录反应将提取的总RNA逆转录为cDNA。C3cDNA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。The extracted total RNA was reverse transcribed into cDNA through reverse transcription reaction. C3cDNA 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.

4.3结果分析4.3 Result analysis

b)使用以下公式计算基因的相对表达量：
ΔCt＝Ct(目的基因)–Ct(GAPDH)
ΔΔCt＝ΔCt(检测样品组)-ΔCt(Mock组)b) Use the following formula to calculate the relative expression of genes:
ΔCt＝Ct(target gene)–Ct(GAPDH)
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–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 sample group)/relative expression of mRNA in the Mock group × 100%.

其中Mock组表示和实验组相比，未加入siRNA缀合物的组。The Mock group represents the group without adding siRNA conjugate compared with the experimental 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.

表11
Table 11

表11中的L96也即式I所示的缀合物基团GalNAc。L96 in Table 11 is also the conjugate group GalNAc shown in Formula I.

从表11中可以看出，本公开提供的siRNA缀合物具有优异的C3基因抑制活性。当通过自由摄取进入PHH细胞时，本公开的siRNA缀合物可在IC₅₀低至0.122nM时仍然具有良好的C3基因抑制活性；当通过转染进入PHH细胞时，可在IC₅₀低至0.013nM时仍然具有良好的C3基因抑制活性。As can be seen from Table 11, the siRNA conjugates provided by the present disclosure have excellent C3 gene inhibition activity. When entering PHH cells through free uptake, the siRNA conjugate of the present disclosure can still have good C3 gene inhibitory activity at an IC ₅₀ as low as 0.122 nM; when entering PHH cells through transfection, the siRNA conjugate can have an IC ₅₀ as low as 0.013 It still has good C3 gene inhibitory activity at nM.

实施例5：siRNA的合成Example 5: Synthesis of siRNA

5.1siRNA序列设计5.1siRNA sequence design

本实施例中的siRNA序列设计思想和方法相同，且用于设计siRNA的靶标序列如下所示，靶标序列来源于C3的基因mRNA序列(参见NM_000064.4)。The siRNA sequence design ideas and methods in this example are the same, and the target sequence used to design siRNA is as follows. The target sequence is derived from the C3 gene mRNA sequence (see NM_000064.4).

靶标序列I:
Target sequence I:

靶标序列I-3:
Target sequence I-3:

5.2合成方法：5.2 Synthesis method:

与实施例1中的“1.2”中的合成方法相同。The same synthesis method as in "1.2" in Example 1.

5.3合成条件：5.3 Synthetic conditions:

与实施例1中的“1.3”中的合成条件相同。The synthesis conditions are the same as in "1.3" in Example 1.

实施例6：siRNA及siRNA修饰物抑制C3基因表达Example 6: siRNA and siRNA modifications inhibit C3 gene expression

6.1实验材料：6.1 Experimental materials:

与实施例3中的“3.1”的实验材料相同。The same experimental materials as “3.1” in Example 3.

6.2实验方法：6.2 Experimental methods:

6.2.1与实施例3中的“3.2.1”中的实验步骤和方法相同。6.2.1 The experimental steps and methods in "3.2.1" in Example 3 are the same.

6.2.2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 siRNA母液。6.2.2 Centrifuge the dry powder of the 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 with ultrapure distilled water (ULtraPure Distilled Water) to prepare 100 μM siRNA. mother liquor.

6.2.3配制2nM的siRNA转染稀释液Z和0.2nM的siRNA转染稀释液W6.2.3 Prepare 2nM siRNA transfection dilution Z and 0.2nM siRNA transfection dilution W

(1)0.1μM siRNA贮备液Q和0.01μM siRNA贮备液E制备：(1) Preparation of 0.1μM siRNA stock solution Q and 0.01μM siRNA stock solution E:

a)、取上述步骤6.2.2中制得的100μM的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 6.2.2 above, and add 18 μl of ultrapure distilled water to obtain a siRNA dilution with a final concentration of 10 μM;

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Q；c). Take 2 μl of the 1 μM siRNA dilution prepared in step b) and add 18 μl of ultrapure distilled water to obtain a siRNA stock solution Q with a final concentration of 0.1 μM;

d)、取步骤c)中制得的0.1μM的siRNA贮备液Q 2μl，加入18μl超纯蒸馏水，得到终浓度为0.01μM的siRNA贮备液E；d), take 2 μl of 0.1 μM siRNA stock solution Q prepared in step c), add 18 μl of ultrapure distilled water, and obtain siRNA stock solution E with a final concentration of 0.01 μM;

(2)取上述配制好的0.1μM siRNA贮备液Q和0.01μM的siRNA贮备液E各2μl，分别加入98μl Opti-medium，得到2nM的siRNA转染稀释液Z和0.2nM的siRNA转染稀释液W。(2) Take 2 μl each of the 0.1 μM siRNA stock solution Q and 0.01 μM siRNA stock solution E prepared above, and add 98 μl Opti-medium respectively to obtain 2 nM siRNA transfection dilution Z and 0.2 nM siRNA transfection dilution. W.

6.2.4转染Hep3B细胞6.2.4 Transfection of Hep3B cells

(1)取Lipofectamine RNAiMax转染试剂3μl，加入97μl Opti-medium，得到Lipofectamine RNAiMax转染试剂稀释液；将Lipofectamine RNAiMax转染试剂稀释液与步骤6.2.3中制得的2nM siRNA稀释液Z以1:1体积比混合，静置5分钟，将10μl的转染混合物加入到96孔板中转染步骤6.2.1中所培养的Hep3B细胞(终体积100μl，该体系中siRNA的浓度为0.1nM)。(1) Take 3 μl of Lipofectamine RNAiMax transfection reagent, add 97 μl of Opti-medium to obtain the Lipofectamine RNAiMax transfection reagent diluent; mix the Lipofectamine RNAiMax transfection reagent diluent and the 2nM siRNA diluent prepared in step 6.2.3 at a ratio of 1 :1 volume ratio, let stand for 5 minutes, add 10 μl of the transfection mixture to the 96-well plate to transfect the Hep3B cells cultured in step 6.2.1 (the final volume is 100 μl, the concentration of siRNA in this system is 0.1 nM) .

(2)取Lipofectamine RNAiMax转染试剂3μl，加入97μl Opti-medium，得到Lipofectamine RNAiMax转染试剂稀释液；将Lipofectamine RNAiMax转染试剂稀释液与步骤6.2.3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6.2.1中培养的Hep3B细胞(终体积100μl，该体系中siRNA的浓度为0.01nM)。(2) Take 3 μl of Lipofectamine RNAiMax transfection reagent and add 97 μl of Opti-medium to obtain Lipofectamine RNAiMax transfection reagent diluent; mix the Lipofectamine RNAiMax transfection reagent diluent with the 0.2nM siRNA diluent prepared in step 6.2.3. Mix 1:1 volume ratio to prepare a transfection mixture, let it stand for 5 minutes, add 10 μl of the transfection mixture to the Hep3B cells cultured in step 6.2.1 in the 96-well plate (the final volume is 100 μl, the concentration of siRNA in this system is 0.01nM).

6.2.5转染后培养48小时，每个浓度设置2个重复。6.2.5 Cultivate for 48 hours after transfection, and set 2 replicates for each concentration.

6.2.6与实施例3中的“3.2.5”的实验步骤和方法相同。6.2.6 The experimental steps and methods of "3.2.5" in Example 3 are the same.

6.2.7对RNA进行逆转录至cDNA，按照以下步骤进行：6.2.7 To reverse transcribe RNA into cDNA, follow the following steps:

a)如下表所述制备逆转录反应混合物并充分混匀，在整个操作过程中，所有试剂都放置在冰上；a) Prepare the reverse transcription reaction mixture as described in the table below and mix thoroughly. During the entire operation, all reagents Place on ice;

表12
Table 12

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.

6.2.8实时PCR分析6.2.8 Real-time PCR analysis

与实施例3中的“3.2.7”的实验步骤和方法相同。The experimental steps and methods are the same as "3.2.7" in Example 3.

6.2.9结果分析6.2.9 Result analysis

与实施例3中的“3.2.8”的分析步骤和方法相同。The analysis steps and methods are the same as "3.2.8" in Example 3.

6.3沉默实验结果6.3 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.

6.3.1设计序列结果6.3.1 Design sequence results

表13抑制率实验结果

Table 13 Inhibition rate experimental results

从表13中可以看出，本公开提供的siRNA、siRNA修饰物显示出对C3基因优异的抑制效果。在0.1nM的浓度下抑制率高达94.07％，在0.01nM的浓度下抑制率高达85.65％。As can be seen from Table 13, the siRNA and siRNA modifications provided by the present disclosure show excellent inhibitory effects on the C3 gene. The inhibition rate is as high as 94.07% at the concentration of 0.1nM, and the inhibition rate is as high as 85.65% at the concentration of 0.01nM.

实施例7：siRNA修饰物及siRNA缀合物抑制C3基因表达Example 7: siRNA modifications and siRNA conjugates inhibit C3 gene expression

7.1实验材料：7.1 Experimental materials:

7.2实验方法：7.2 Experimental methods:

siRNA修饰物(siRNA修饰物的终浓度包括2nM和0.2nM两组，每个抑制实验均设置复孔)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Lipofectamine RNAiMax将siRNA修饰物转入细胞，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RNA提取。根据说明书使用96Kit提取总RNA。siRNA modifications (the final concentrations of siRNA modifications include two groups of 2nM and 0.2nM, and duplicate wells are set for each inhibition experiment) are entered into PHH cells through transfection. The process is as follows: take the frozen PHH cells, recover, and count. Adjust the cells to 6×10 ⁵ cells/ml, and use Lipofectamine RNAiMax to transfer the siRNA modification into the cells. Seed into a 96-well plate at a density of 54,000 cells per well, and add 100 μL of PPH medium 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 RNA extraction. Use according to instructions 96Kit extracted total RNA.

siRNA缀合物(siRNA缀合物终浓度包括200nM和10nM两组，每个抑制实验均设置复孔)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，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RNA提取。根据说明书使用96Kit提取总RNA。siRNA conjugates (the final concentrations of siRNA conjugates include two groups of 200nM and 10nM, and multiple wells are set for each inhibition experiment) enter PHH cells through free uptake. The process is as follows: take the frozen PHH cells, recover, and count. Adjust the cells to 6 × 10 ⁵ cells/ml, add siRNA conjugate, and seed 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 RNA extraction. Use according to instructions 96Kit extracted total RNA.

7.3结果分析7.3 Result analysis

其中Mock组表示和实验组相比，未加入siRNA缀合物和siRNA修饰物的组。The Mock group represents the group in which siRNA conjugates and siRNA modifications were not added compared with the experimental group.

表14

Table 14

表15

Table 15

从表14和15中可以看出，本公开的siRNA修饰物、siRNA缀合物具有很高的C3基因抑制活性。As can be seen from Tables 14 and 15, the siRNA modifications and siRNA conjugates of the present disclosure have high C3 gene inhibitory activity.

实施例8 siRNA缀合物在人源化小鼠中对人C3基因表达的抑制作用Example 8 Inhibitory effect of siRNA conjugates on human C3 gene expression in humanized mice

6-8周龄的C57BL/6-hC3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siRNA193、siRNA194和siRNA197(每组6只小鼠)。给药后第7日、第14日、第21日、第28日和第35日检测血清hC3蛋白表达量，由此得出siRNA缀合物对hC3蛋白表达的抑制率。C57BL/6-hC3 mice (provided by Shanghai Southern Model Biotechnology Co., Ltd.) aged 6-8 weeks entered the breeding facility. After adaptive feeding for 7 days, siRNA193 was administered subcutaneously to the mice at a single dose of 3 mg/kg. , siRNA194 and siRNA197 (6 mice per group). The serum hC3 protein expression was detected on the 7th, 14th, 21st, 28th and 35th days after administration, and the inhibition rate of hC3 protein expression by the siRNA conjugate was obtained.

表16 siRNA缀合物对hC3蛋白抑制率
Table 16 Inhibition rate of hC3 protein by siRNA conjugates

从表16可以看出，本公开的siRNA缀合物在体内对hC3基因具有较高的抑制活性，能够长时间降低hC3蛋白水平，剂量效应明显。具体而言，在单次皮下给药3mg/kg后，在第7天时，siRNA193对hC3基因显示了高达67.2％的抑制；在第35天时，siRNA193对hC3基因仍然显示了48.3％的抑制率；siRNA194在第7天时对hC3基因显示了高达 73.3％的抑制；在第35天时，siRNA194对hC3基因显示了55.7％的抑制；siRNA197在第7天时对hC3基因显示了高达64.4％的抑制；在第35天时，siRNA197对hC3基因显示了49.1％的抑制。As can be seen from Table 16, the siRNA conjugate of the present disclosure has high inhibitory activity on the hC3 gene in vivo, can reduce hC3 protein levels for a long time, and has an obvious dose effect. Specifically, after a single subcutaneous administration of 3 mg/kg, siRNA193 showed up to 67.2% inhibition of the hC3 gene on day 7; on day 35, siRNA193 still showed 48.3% inhibition of the hC3 gene; siRNA194 showed up to 73.3% inhibition; on day 35, siRNA194 showed 55.7% inhibition of hC3 gene; siRNA197 showed up to 64.4% inhibition of hC3 gene on day 7; on day 35, siRNA197 showed 49.1% inhibition of hC3 gene of inhibition.

实施例9 siRNA缀合物在CD-1小鼠中的血浆动力学研究Example 9 Plasma kinetics study of siRNA conjugates 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：siRNA缀合物在3mg/kg(10mL/kg)的剂量下给药，随机分组后单次皮下注射给药，每组6只小鼠。Dosage and mode of administration: siRNA conjugate was administered at a dose of 3 mg/kg (10 mL/kg), and administered as a single subcutaneous injection after random grouping, with 6 mice in each 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：C_max、T_max、AUC、MRT、t_1/2。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

给药剂量和方式：siRNA缀合物在3mg/kg(10mL/kg)的剂量下给药，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 mode of administration: siRNA conjugate was administered at a dose of 3 mg/kg (10 mL/kg), and administered as a single subcutaneous injection after randomization, with 3 animals at each time point, for a total of 24 mice.

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；168h (1 week) after administration: collect plasma, liver, kidney, spleen, brain, heart, lung, stomach, small intestine, muscle, and testis;

给药后336h(2周)：采集血浆、肝、肾、脾；336h (2 weeks) after administration: plasma, liver, kidney, spleen are collected;

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C57小鼠给予MTD试验Example 11 Single subcutaneous injection of siRNA conjugate into C57 mice for MTD test

C57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：

C57 mice, SPF grade, male, about 25 g, 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 as follows:

检测指标：Detection Indicator:

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Clinical observation: Observe continuously for 4 hours on the dosing day, and perform clinical observation at least once a day during the recovery period.

体重：对所有存活动物每周进行2次体重称量。Body weight: All surviving animals were weighed twice a week.

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)。Immunotoxicity: Animals in the MTD dose group were blood collected 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).

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。Toxicokinetics: Animals in the MTD dose group were blood collected alternately before D1 administration and 30min±2min, 1h±2min, 4h±5min, 8h±10min, and 24h±20min after D1 administration. 3 animals/sex/ were collected at each time point. Group animals and detect blood drug concentrations.

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。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.

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。Tissue distribution: 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. Blood and liver were collected to detect tissue drug concentration.

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。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 abnormal tissues were collected Or organs were harvested 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

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;

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;

The target sequence is selected from the nucleotide sequence shown in any one of SEQ ID NO: 1-7.

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:8-34 and SEQ ID NO:364, and the sense strand includes as Sequence A consisting of at least 15 consecutive nucleotides in the nucleotide sequence shown in any one of SEQ ID NO:8-34 and SEQ ID NO:364, and the antisense strand includes SEQ ID NO:8-34 , a reverse complementary and/or substantially reverse complementary sequence B consisting of at least 15 consecutive nucleotides in the nucleotide sequence shown in any one of SEQ ID NO: 364.

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.

The double-stranded ribonucleic acid according to claim 3, wherein the nucleotide sequence of the sense strand is 15 of the nucleotide sequences shown in any one of SEQ ID NO:8-34 and SEQ ID NO:364 -Sequence A consisting of 28 consecutive nucleotides, preferably 19-25 consecutive nucleotides, more preferably 19-23 consecutive nucleotides, more preferably 19, 21 or 23 nucleotides.

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.

The double-stranded ribonucleic acid according to claim 5, wherein the nucleotide sequence of the antisense strand is the same as the nucleotide sequence shown in any one of SEQ ID NO:8-34 and SEQ ID NO:364. The sequence B consisting of 15-28 consecutive nucleotides is reverse complementary and/or substantially reverse complementary, preferably 19-25 consecutive nucleotides, more preferably 19-23 consecutive nucleotides, more preferably 19, 21 or 23 nucleotides.

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.

The double-stranded ribonucleic acid according to any one of claims 1-7, wherein,

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,

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,

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,

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.

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:

1) 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:131;

2) 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:132;

3) 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:133;

4) 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:134;

5) 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:135;

6) 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:136;

7) 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:137;

8) 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:138;

9) 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:139;

10) 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:140;

11) 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:141;

12) 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:142;

13) 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:143;

14) 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:144;

15) 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:145;

16) 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:146;

17) 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:147;

18) 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:148;

19) 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:149;

20) 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:150;

21) 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:151;

22) 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:152;

23) 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:153;

24) 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:154;

25) 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:155;

26) 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:156;

27) 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:157;

28) 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. The nucleotide sequence shown in NO:158;

29) 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:159;

30) 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:160;

31) 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:161;

32) 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:162;

33) 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: 163;

34) 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:164;

35) 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:165;

36) 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:166;

37) 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:167;

38) 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:168;

39) 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:169;

40) 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:170;

41) 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:171;

42) 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:172;

43) 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:173;

44) 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:174;

45) 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:175;

46) 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:176;

47) 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:177;

48) 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:178;

49) 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:179;

50) 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:180;

51) 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:181;

52) 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:182;

53) 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:183;

54) 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:184;

55) 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:185;

56) 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:186;

57) 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:187;

58) 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:188;

59) 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:189;

60) 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:190;

61) 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:191;

62) 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:192;

63) 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:193;

64) 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:194;

65) 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:195;

66) 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:196;

67) 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:197;

68) 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:198;

69) 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:199;

70) 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:200;

71) 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:201;

72) 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:202;

73) 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:107 The nucleotide sequence shown in NO:203;

74) 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:204;

75) 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:205;

76) 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:206;

77) 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:207;

78) 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:208;

79) 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:209;

80) 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:210;

81) 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:211;

82) 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:212;

83) 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:213;

84) 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:214;

85) 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:215;

86) 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:216;

87) The sense strand includes the nucleotide sequence shown in SEQ ID NO:121, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:217;

88) The sense strand includes the nucleotide sequence shown in SEQ ID NO:122, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:218;

89) The sense strand includes the nucleotide sequence shown in SEQ ID NO:123, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:219;

90) The sense strand includes the nucleotide sequence shown in SEQ ID NO:124, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:220;

91) The sense strand includes the nucleotide sequence shown in SEQ ID NO:125, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:221;

92) The sense strand includes the nucleotide sequence shown in SEQ ID NO:126, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:222;

93) The sense strand includes the nucleotide sequence shown in SEQ ID NO:127, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:223;

94) The sense strand includes the nucleotide sequence shown in SEQ ID NO:128, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:224;

95) The sense strand includes the nucleotide sequence shown in SEQ ID NO:129, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:225;

118) The sense strand includes the nucleotide sequence shown in SEQ ID NO:328, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:346;

119) The sense strand includes the nucleotide sequence shown in SEQ ID NO:329, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:347;

120) The sense strand includes the nucleotide sequence shown in SEQ ID NO:330, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:348;

121) The sense strand includes the nucleotide sequence shown in SEQ ID NO:331, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:349;

122) The sense strand includes the nucleotide sequence shown in SEQ ID NO:332, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:350;

123) The sense strand includes the nucleotide sequence shown in SEQ ID NO:333, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:351;

124) The sense strand includes the nucleotide sequence shown in SEQ ID NO:334, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:352;

125) The sense strand includes the nucleotide sequence shown in SEQ ID NO:335, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:353;

126) The sense strand includes the nucleotide sequence shown in SEQ ID NO:336, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:354;

127) The sense strand includes the nucleotide sequence shown in SEQ ID NO:337, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:355;

128) The sense strand includes the nucleotide sequence shown in SEQ ID NO:338, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:356;

129) The sense strand includes the nucleotide sequence shown in SEQ ID NO:339, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:357;

130) The sense strand includes the nucleotide sequence shown in SEQ ID NO:340, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:358;

131) The sense strand includes the nucleotide sequence shown in SEQ ID NO:341, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:359;

132) The sense strand includes the nucleotide sequence shown in SEQ ID NO:342, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:360;

133) The sense strand includes the nucleotide sequence shown in SEQ ID NO:343, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:361;

134) 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:362;

135) 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:363.

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.

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.

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.

The double-stranded ribonucleic acid according to any one of claims 1-12, wherein the double-stranded ribonucleic acid is 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 C3 gene expression.

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) Modification of at least one nucleotide in the sense strand,

(2) Modification of the phosphodiester bond at at least one position in the sense strand,

(3) Modification of at least one nucleotide in the antisense strand,

(4) Modification of the phosphodiester bond at at least one position in the antisense strand;

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;

Alternatively, the sense strand and antisense strand of the double-stranded ribonucleic acid modification are selected from the following sequence combinations:

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, 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;

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;

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;

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;

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.

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.

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.

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.

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.

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:

Between the first nucleotide and the second nucleotide starting from the 5' end of the sense strand;

Between the 2nd and 3rd nucleotide starting from the 5' end of the sense strand;

Between the first nucleotide and the second nucleotide starting from the 3' end of the sense strand;

Between the second and third nucleotides starting from the 3' end of the sense strand;

or,

The sense strand contains phosphorothioate diester bonds at the positions shown below:

Between the 2nd and 3rd nucleotide starting from the 5' end of the sense strand.

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;

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;

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;

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;

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.

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.

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:

Between the first nucleotide and the second nucleotide starting from the 5' end of the antisense strand;

Between the 2nd and 3rd nucleotide starting from the 5' end of the antisense strand;

Between the first nucleotide and the second nucleotide starting from the 3' end of the antisense strand;

Between the 2nd and 3rd nucleotide starting from the 3' end of the antisense strand.

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 ) :

(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',

(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',

(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',

(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',

(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';

Wherein, N ₁ -N ₂₃ are independently selected from ribonucleotides whose bases are A, U, C or G,

The capital letter T stands for deoxyribonucleotide whose base is thymine.

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.

The lowercase letter f indicates that the ribonucleotide adjacent to the left side of the letter f is a 2’-F modified ribonucleotide.

-(s)- indicates that two adjacent nucleotides are connected by a phosphorothioate diester bond.

The double-stranded ribonucleic acid modification according to any one of claims 15-24, wherein the double-stranded ribonucleic acid The antisense strand of the acid modification has a structure shown in any one of (b ₁ ) to (b ₁₅ ):

(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',

(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',

(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',

(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'，

(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',

(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',

(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'，

(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',

(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',

(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',

(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',

(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' ,

(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',

(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',

(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 ';

The capital letter T stands for deoxyribonucleotide whose base is thymine.

P1 indicates that the nucleotide adjacent to the right side of the letter is a 5’-phosphate nucleotide,

-(s)- means that two adjacent nucleotides are connected by a phosphorothioate diester bond,

(GNA) indicates that the ribonucleotide adjacent to the right side is a ribonucleotide modified with GNA.

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.

The double-stranded ribonucleic acid modification product according to any one of claims 15 to 26, wherein the double-stranded ribonucleic acid modification product is an siRNA modification product used to inhibit C3 gene expression.

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:

97) 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:227 The nucleotide sequence shown in NO:242;

98) 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:243;

99) 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:244;

100) 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:245;

101) 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:246;

102) 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:247;

103) 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:248;

104) 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:249;

105) 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:250;

106) 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:251;

107) The sense strand includes the nucleotide sequence shown in SEQ ID NO:237, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:252;

108) The sense strand includes the nucleotide sequence shown in SEQ ID NO:238, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:253;

109) The sense strand includes the nucleotide sequence shown in SEQ ID NO:239, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:254;

110) The sense strand includes the nucleotide sequence shown in SEQ ID NO:240, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:255;

111) The sense strand includes the nucleotide sequence shown in SEQ ID NO:241, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:256;

115) 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:260;

116) 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:261;

117) 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:262;

136) 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:283;

137) 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:283;

138) 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:284;

139) 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:285;

140) 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:286;

141) 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:287;

142) 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:287;

143) 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:288;

144) 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:289;

145) 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:290;

146) 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:291;

147) 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:291;

148) 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:292;

149) 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:293;

150) 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:294;

151) The sense strand includes the nucleotide sequence shown in SEQ ID NO:238, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:295;

152) 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:295;

153) The sense strand includes the nucleotide sequence shown in SEQ ID NO:238, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:296;

154) The sense strand includes the nucleotide sequence shown in SEQ ID NO:238, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:297;

155) The sense strand includes the nucleotide sequence shown in SEQ ID NO:238, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:298;

156) The sense strand includes the nucleotide sequence shown in SEQ ID NO:268, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:299;

157) The sense strand includes the nucleotide sequence shown in SEQ ID NO:269, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:299;

158) The sense strand includes the nucleotide sequence shown in SEQ ID NO:268, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:300;

159) The sense strand includes the nucleotide sequence shown in SEQ ID NO:268, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:301;

160) The sense strand includes the nucleotide sequence shown in SEQ ID NO:268, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:302;

161) The sense strand includes the nucleotide sequence shown in SEQ ID NO:270, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:303;

162) The sense strand includes the nucleotide sequence shown in SEQ ID NO:271, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:303;

163) The sense strand comprises the nucleotide sequence shown in SEQ ID NO:270, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:270 The nucleotide sequence shown in NO:304;

164) The sense strand includes the nucleotide sequence shown in SEQ ID NO:270, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:305;

165) The sense strand includes the nucleotide sequence shown in SEQ ID NO:270, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:306;

166) The sense strand includes the nucleotide sequence shown in SEQ ID NO:272, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:307;

167) The sense strand includes the nucleotide sequence shown in SEQ ID NO:273, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:307;

168) The sense strand includes the nucleotide sequence shown in SEQ ID NO:272, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:308;

169) The sense strand includes the nucleotide sequence shown in SEQ ID NO:272, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:309;

170) The sense strand includes the nucleotide sequence shown in SEQ ID NO:272, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:310;

171) The sense strand includes the nucleotide sequence shown in SEQ ID NO:274, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:311;

172) 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:311;

173) The sense strand includes the nucleotide sequence shown in SEQ ID NO:274, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:312;

174) The sense strand includes the nucleotide sequence shown in SEQ ID NO:274, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:313;

175) The sense strand includes the nucleotide sequence shown in SEQ ID NO:274, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:314;

176) The sense strand includes the nucleotide sequence shown in SEQ ID NO:275, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:315;

177) The sense strand includes the nucleotide sequence shown in SEQ ID NO:276, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:316;

178) The sense strand includes the nucleotide sequence shown in SEQ ID NO:277, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:317;

179) 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:318;

180) 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:319;

181) 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:320;

182) 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:321;

183) 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:322.

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.

The double-stranded ribonucleic acid conjugate according to claim 29, wherein the conjugation group has the following structure:

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.

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;

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;

or,

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.

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:

Among them, the double helix structure is double-stranded ribonucleic acid or double-stranded ribonucleic acid modification.

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.

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 C3 gene expression.

The double-stranded ribonucleic acid conjugate according to any one of claims 29 to 35, wherein in the double-stranded ribonucleic acid conjugate, the sense strand and antisense strand of the double-stranded ribonucleic acid are selected from the following combinations :

4) 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:38 The nucleotide sequence shown in NO:134;

28) 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:158;

49) 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:83 The nucleotide sequence shown in NO:179;

73) 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:203;

94) The sense strand includes the nucleotide sequence shown in SEQ ID NO:128, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO. The nucleotide sequence shown in NO:224;

135) 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:363;

97) 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:242;

143) The sense strand comprises the nucleotide sequence shown in SEQ ID NO:236, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:236 The nucleotide sequence shown in NO:288;

163) The sense strand includes the nucleotide sequence shown in SEQ ID NO:270, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:304;

183) 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:322;

Optionally, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:257, and the antisense strand includes the sequence shown in SEQ ID NO:260 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:258, and the antisense strand includes the core shown in SEQ ID NO:261 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:259, and the antisense strand includes the core shown in SEQ ID NO:262 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:257, and the antisense strand includes the core shown in SEQ ID NO:283 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes SEQ ID NO: 257 The sequence shown, the antisense strand includes the nucleotide sequence shown in SEQ ID NO:284;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:257, and the antisense strand includes the core shown in SEQ ID NO:285 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:257, and the antisense strand includes the core shown in SEQ ID NO:286 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:323, and the antisense strand includes the core shown in SEQ ID NO:251 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:323, and the antisense strand includes the core shown in SEQ ID NO:287 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO: 323, and the antisense strand includes the core shown in SEQ ID NO: 288 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:323, and the antisense strand includes the core shown in SEQ ID NO:289 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:323, and the antisense strand includes the core shown in SEQ ID NO:290 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:258, and the antisense strand includes the core shown in SEQ ID NO:291 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:258, and the antisense strand includes the core shown in SEQ ID NO:292 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:258, and the antisense strand includes the core shown in SEQ ID NO:293 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:258, and the antisense strand includes the core shown in SEQ ID NO:294 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:259, and the antisense strand includes the core shown in SEQ ID NO:295 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:259, and the antisense strand includes the core shown in SEQ ID NO:296 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:259, and the antisense strand includes the core shown in SEQ ID NO:297 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:259, and the antisense strand includes the core shown in SEQ ID NO:298 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:324, and the antisense strand includes the core shown in SEQ ID NO:299 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:324, and the antisense strand includes the core shown in SEQ ID NO:300 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:324, and the antisense strand includes the core shown in SEQ ID NO:301 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:324, and the antisense strand includes the core shown in SEQ ID NO:302 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:325, and the antisense strand includes the core shown in SEQ ID NO:303 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:325, and the antisense strand includes the core shown in SEQ ID NO:304 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:325, and the antisense strand includes the core shown in SEQ ID NO:305. nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:325, and the antisense strand includes the core shown in SEQ ID NO:306 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:326, and the antisense strand includes the core shown in SEQ ID NO:307 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:326, and the antisense strand includes the core shown in SEQ ID NO:308 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:326, and the antisense strand includes the core shown in SEQ ID NO:309 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:326, and the antisense strand includes the core shown in SEQ ID NO:310 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:327, and the antisense strand includes the core shown in SEQ ID NO:311 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:327, and the antisense strand includes the core shown in SEQ ID NO:312 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:327, and the antisense strand includes the core shown in SEQ ID NO:313 nucleotide sequence;

Alternatively, in the double-stranded ribonucleic acid conjugate, the sense strand connecting the conjugate group includes the sequence shown in SEQ ID NO:327, and the antisense strand includes the core shown in SEQ ID NO:314. nucleotide sequence.

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.

The pharmaceutical composition according to claim 37, wherein the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers.

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) Inhibit C3 gene expression, or prepare drugs for inhibiting C3 gene expression;

(2) For preventing or treating diseases related to abnormal expression of C3 gene, or preparing drugs for preventing or treating diseases related to abnormal expression of C3 gene;

(3) For use in treating subjects suffering from a disease that would benefit from reduced expression of the complement C3 gene, or to prepare a medicament for treating subjects suffering from a disease that would benefit from reduced expression of the complement C3 gene.

The use according to claim 39, wherein the disease associated with abnormal expression of the C3 gene is selected from the group consisting of the following diseases:

Paroxysmal nocturnal hemoglobinuria, atypical hemolytic uremic syndrome, rheumatoid arthritis, chronic kidney disease, ischemia-reperfusion injury, neurodegenerative diseases, asthma, systemic lupus erythematosus, glomerulonephritis, Psoriasis, dermatomyositis bullous pemphigoid, Shiga toxin E. coli-related hemolytic uremic syndrome, myasthenia gravis, neuromyelitis optica, dense deposit disease, C3 neurological disease, age-related macular degeneration, cold agglutinin disease, antineutrophil cytoplasmic antibody-associated vasculitis, humoral and vascular graft rejection, graft dysfunction, myocardial infarction, sensitized recipients of grafts, and sepsis.

A method for inhibiting intracellular C3 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.

The method of claim 41, wherein the cells are in vivo cells or in vitro cells.

The method of claim 41 or 42, wherein the cells are in the subject.

The method of claim 43, wherein the subject is a mammal, preferably a human.

The method of claim 43 or 44, wherein the subject has at least one of the following characteristics:

Abnormal expression of C3 gene in the body, more specifically abnormal high expression of C3 gene;

Suffering from diseases related to abnormal expression of C3 gene;

People with diseases that would benefit from reduced C3 gene expression.