Rnai reagent targeting complement c3 and use thereof

Abstract

Provided are an RNAi reagent targeting complement C3 and a use thereof. The RNAi reagent can effectively inhibit the expression of complement C3 in cells, and thus can be used for treating diseases related to complement C3.

Description

RNAi agents targeting c3 complement and uses thereof

[0001] Cross-reference to Related Applications

[0002] This application claims priority to Chinese Patent Application No. 2024111277755, filed on August 16, 2024, the entire contents of which are incorporated herein by reference. TECHNICAL FIELD

[0003] The present application belongs to the field of biological medicine, and specifically relates to RNAi agents targeting C3 complement and uses thereof. BACKGROUND

[0004] The complement system is an immune defense system of the human body composed of more than 30 proteins (Janeway, CA Jr et al, 2001). There are mainly three complement activation pathways, the classical pathway, the alternative pathway, and the lectin pathway (Abbas AK et al, 2010). Complement activation leads to a series of linked enzymatic reactions, forming anaphylatoxins C3a and C5a and further triggering downstream physiological responses.

[0005] The classical pathway is activated through the binding of C1 to the classical pathway activator (mainly an antigen-antibody complex containing IgM, IgG1, IgG2, or IgG3). The reaction sequence of the classical complement activation pathway is: C1, 4, 2, 3, 5, 6, 7, 8, 9. This biochemical pathway forms a pore on the cell membrane, leading to swelling and rupture of the target cell due to osmotic pressure.

[0006] The activation of the alternative pathway depends on the natural hydrolysis of C3 into C3a and C3b, and C3b binds to factors B, D, and P to proceed similarly to the reactions of the classical pathway.

[0007] The full name of the lectin pathway is the mannose-binding lectin pathway. Mannose-binding lectin can recognize the mannose residues on the surface of pathogens, and then bind to the surface of pathogens and activate reactions similar to the classical pathway.

[0008] Aberrant complement activation can lead to a series of pathological changes in many diseases, such as paroxysmal nocturnal hemoglobinuria (PNH), generalized myasthenia gravis (GMG), rheumatoid arthritis (RA) and neuromyelitis optica (NMO) and the like.

[0009] C3 complement (Complement Component 3, also known as Complement C3 or Complement Component 3) plays an important role in the whole complement system, and the classical activation pathway and the alternative activation pathway both depend on the activation of C3 (Lambris 2001). C3 is also the most abundant complement in human blood, mainly produced in the liver (de Bruijn MH and Fey GH, 1985). Currently, there is only one monoclonal antibody Pegacetacoplan targeting C3 complement, which is expensive and needs frequent injection.

[0010] RNA interference (RNAi) refers to a highly conserved phenomenon in the evolution process, which is induced by double-stranded small interfering RNA (siRNA) to efficiently and specifically degrade homologous mRNA. At the same time, RNAi drugs also have the advantage of long drug effect time compared with antibodies. Therefore, it is of great significance to research and develop siRNA targeting C3 complement. SUMMARY

[0011] The first object of the present application is to provide an RNAi agent, which comprises: an antisense strand containing at least 15 consecutive nucleotides, and the difference between the at least 15 consecutive nucleotides and any of the at least 15 consecutive nucleotides in any of SEQ ID NOs: 54-106 is not more than 3 nucleotides; and a sense strand containing a nucleotide sequence complementary to at least part of the antisense strand.

[0012] The present application also provides an ex vivo cell containing the RNAi agent.

[0013] The present application also provides a pharmaceutical composition comprising the RNAi agent.

[0014] The second object of the present application is to provide a method for inhibiting the expression of C3 complement in a cell, which comprises: contacting the cell with the RNAi agent or the pharmaceutical composition to inhibit the expression of C3 complement in the cell.

[0015] The present application also provides the use of the RNAi agent or the pharmaceutical composition in treating and / or preventing a disease associated with C3 complement.

[0016] The present application also provides the use of the RNAi agent or the pharmaceutical composition in the preparation of a medicament for treating and / or preventing a disease associated with C3 complement.

[0017] The RNAi agent of the present application can effectively inhibit the expression of C3 complement in cells, and thus can be used for treating a disease associated with C3 complement. BRIEF DESCRIPTION OF DRAWINGS

[0018] In order to more clearly illustrate the specific embodiments of the present application or the technical solutions in the prior art, the drawings needed in the specific embodiments or prior art description will be briefly introduced as follows. Obviously, the drawings in the following description are some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative labor.

[0019] Figure 1 is the drug efficacy of different siRNAs in human C3 transgenic mice in Example 3 of the present application.

[0020] Figure 2 is the long-acting inhibition result of PAT-0038T in human C3 transgenic mice in Example 5 of the present application. DETAILED DESCRIPTION

[0021] The specific embodiments of the present application will be described in detail below. It should be understood that the specific embodiments described herein are only for the purpose of illustration and explanation of the present application, and are not intended to limit the present application, and those skilled in the art can make various modifications and changes to the present application without departing from the scope or spirit of the present application. For example, features described or illustrated as part of one embodiment can be used in another embodiment to produce further embodiments.

[0022] Unless otherwise defined, all terms (including technical and scientific terms) used in the disclosure of the present application have the same meaning as commonly understood by those skilled in the art to which the present application belongs. For further guidance, the following definitions are used to better understand the teachings of the present application. The terms used in the specification of the present application herein are only for the purpose of describing the specific embodiments and are not intended to limit the present application.

[0023] The selection scope of the terms "and / or", "or / and", "and / or" used herein includes any one of the two or more related listed items, and also includes any and all combinations of the related listed items, including any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least two conjunctions selected from "and / or", "or / and", "and / or" are combined to connect at least three items, it should be understood that in this application, the technical solution undoubtedly includes the technical solution connected by "logical and", and also undoubtedly includes the technical solution connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For another example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C and D (i.e. the technical solution connected by "logical or"), and also includes any and all combinations of A, B, C and D, i.e. includes the combination of any two or any three of A, B, C and D, and also includes the four-item combination of A, B, C and D (i.e. the technical solution connected by "logical and").

[0024] The terms "containing", "including" and "comprising" used in the present application are synonymous and are inclusive or open-ended, and do not exclude additional, unrecited members, elements or method steps.

[0025] The numerical ranges used in the present application include all the numerical values and fractions contained in the range, as well as the recited endpoints.

[0026] In the present application, the concentration values involved include fluctuations within a certain range. For example, it can fluctuate within a corresponding accuracy range. For example, 2% can be allowed to fluctuate within ±0.1%. For larger values or values that do not need to be controlled too finely, it is also allowed that the meaning includes larger fluctuations. For example, 80% can be allowed to fluctuate within ±1%, ±2%, ±5%, etc.

[0027] In the present application, the descriptions such as "multiple", "various" and the like refer to greater than or equal to 2 in quantity, unless otherwise specified.

[0028] In the present application, the technical features described in an open-ended manner include both the closed technical solution consisting of the listed features, and the open technical solution containing the listed features.

[0029] In the present application, "preferably", "better", "better", "suitable" only describe the implementation manner or embodiment with better effect, and it should be understood that it does not constitute a limitation on the protection scope of the present application.

[0030] In the present application, "optionally", "optional", "option", "alternatively", "alternative", "alternatively" means optional, that is, selected from "yes" or "no" two parallel schemes. If there are multiple "options" or "alternatives" in a technical solution, if there is no special description, and there is no contradiction or mutual restriction, each "option" or "alternative" is independent.

[0031] In the present application, the term "RNAi agent" (also referred to as "RNAi trigger") means a composition containing an RNA or RNA-like (e.g., chemically modified RNA) oligonucleotide molecule that is capable of degrading or inhibiting (e.g., under appropriate conditions) the translation of a messenger RNA (mRNA) transcript of a target mRNA in a sequence-specific manner. As used herein, an RNAi agent can act through an RNA interference mechanism (i.e., by interacting with the RNA interference pathway machinery of a mammalian cell (the RNA-induced silencing complex or RISC) to induce RNA interference), or through any alternative mechanism or pathway. While it is believed that, as the term is used herein, an RNAi agent acts primarily through an RNA interference mechanism, the disclosed RNAi agents are not bound or limited to any particular pathway or mechanism of action. The RNAi agents disclosed herein are composed of a sense strand and an antisense strand, and include, but are not limited to, short (or small) interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), short hairpin RNA (shRNA), and dicer substrates. The antisense strand of the RNAi agents described herein is at least partially complementary to the mRNA to be targeted (i.e., the C3 complement mRNA). The RNAi agents can include one or more modified nucleotides and / or one or more non-phosphodiester linkages.

[0032] In the present application, when referring to an "antisense (sense) strand comprising at least 15 contiguous nucleotides, and said at least n contiguous nucleotides differ from at least 15 contiguous nucleotides of any of the reference sequences (such as the sequence set forth in SEQ ID No. X or the nucleotide sequence set forth in positions 1-21 of the sequence) by no more than 3 nucleotides", the alignment context involved includes alignment with at least 15 contiguous nucleotides starting at any position (such as position 1, 2, 3,..., 7, or 8) of the reference sequence. As an example, in some embodiments, the antisense (sense) strand comprises 21 contiguous nucleotides, wherein the nucleotide sequence in positions 1-15 differs from the nucleotide sequence in positions 2-16 of the reference sequence by 1, 2, or 3 differences, and the nucleotide sequence in positions 16-20 of the antisense (sense) strand is identical to or different from the nucleotide sequence in positions 17-21 of the reference sequence, such a sequence is also within the scope of the present application. In some embodiments, the antisense (sense) strand comprises 21 contiguous nucleotides, wherein the nucleotide sequence in positions 1-15 is identical to the nucleotide sequence in positions 1-15 of the reference sequence, and the nucleotide sequence in positions 16-21 of the antisense (sense) strand is identical to or different from the nucleotide sequence in positions 16-21 of the reference sequence, such a sequence is also within the scope of the present application.

[0033] In the present application, when referring to descriptions related to sequence alignment, the term "differ" or "differences" includes one or several of the following situations: substitution, insertion, deletion.

[0034] In the present application, when referring to expression of a given gene, the terms "silencing", "reducing", "inhibiting", "down-regulating" or "knocking down" mean that the expression of the gene is reduced when a cell, a population of cells, a tissue, an organ or a subject is treated with an RNAi agent described herein, as measured by the level of RNA transcribed from the gene or the level of polypeptide, protein or protein subunit translated from mRNA in the cell, population of cells, tissue, organ or subject in which the gene is transcribed, compared to a second cell, population of cells, tissue, organ or subject not so treated.

[0035] In the present application, "fully complementary" means that in a hybridizing pair of nucleobase or nucleotide sequence molecules, all (100%) of the bases in the contiguous sequence of a first oligonucleotide hybridize with the same number of bases in the contiguous sequence of a second oligonucleotide. The contiguous sequence can comprise all or a portion of the first or second nucleotide sequence.

[0036] In the present invention, "partially complementary" means that, in a hybridizing pair of nucleobase or nucleotide sequence molecules, at least 70% but not all of the bases in the contiguous sequence of a first oligonucleotide hybridize with the same number of bases in the contiguous sequence of a second oligonucleotide. The contiguous sequence can comprise all or a portion of the first or second nucleotide sequence.

[0037] In the present invention, "substantially complementary" means that, in a hybridizing pair of nucleobase or nucleotide sequence molecules, at least 85% but not all of the bases in the contiguous sequence of a first oligonucleotide hybridize with the same number of bases in the contiguous sequence of a second oligonucleotide. The contiguous sequence can comprise all or a portion of the first or second nucleotide sequence.

[0038] In the present invention, when referring to "at least partially complementary" means that, in a hybridizing pair of nucleobase or nucleotide sequence molecules, a first oligonucleotide is partially complementary, substantially complementary, or fully complementary to a second oligonucleotide.

[0039] In the present invention, the term "treatment" means a method or procedure taken to provide relief or reduction in the number, severity, and / or frequency of one or more symptoms of a disease in a subject. The treatment can include prevention, management, prophylactic treatment, and / or inhibition or reduction in the number, severity, and / or frequency of one or more symptoms of a disease in a subject.

[0040] In the present invention, the term "linked" means that two compounds or molecules are joined by a covalent bond. Unless otherwise specified, as used herein, the term "linked" can refer to a linkage between a first compound and a second compound with or without any intervening atoms or groups of atoms.

[0041] RNAi agent

[0042] The present invention first provides an RNAi agent comprising: an antisense strand containing at least 15, 16, 17, 18, 19, 20, 21, 22, or 23 contiguous nucleotides, and the at least 15, 16, 17, 18, 19, 20, 21, 22, or 23 contiguous nucleotides differ from any of at least 15, 16, 17, 18, 19, 20, 21, 22, or 23 contiguous nucleotides in any of SEQ ID NOs: 54-106 by no more than 0, 1, 2, or 3 nucleotides; and a sense strand comprising a nucleotide sequence at least partially complementary (e.g., partially complementary, substantially complementary, or fully complementary) to the antisense strand.

[0043] In some embodiments, the sense strand contains at least 15, 16, 17, 18, 19, 20, or 21 contiguous nucleotides, and the at least 15, 16, 17, 18, 19, 20, or 21 contiguous nucleotides differ by no more than 0, 1, 2, or 3 nucleotides when compared to any 15, 16, 17, 18, 19, 20, or 21 contiguous nucleotides of any of the sequences set forth in SEQ ID Nos. 1-53.

[0044] In some embodiments, the antisense strand has 15-30 (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides (bases).

[0045] In some embodiments, the sense strand has 15-30 (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides (bases).

[0046] In the present application, the sense strand and the antisense strand can have the same length, or can have different lengths.

[0047] In practice, one skilled in the art can combine the sequences provided in the present application in consideration of the complementarity of the sense strand and the antisense strand, to obtain the combined RNAi agent (e.g., siRNA).

[0048] In a preferred embodiment of the present application, the RNAi agent is selected from the group consisting of siRNA-1 with sense sequence of SEQ ID NO: 1 and antisense sequence of SEQ ID NO: 54, siRNA-2 with sense sequence of SEQ ID NO: 2 and antisense sequence of SEQ ID NO: 55, siRNA-3 with sense sequence of SEQ ID NO: 3 and antisense sequence of SEQ ID NO: 56, siRNA-4 with sense sequence of SEQ ID NO: 4 and antisense sequence of SEQ ID NO: 57, siRNA-5 with sense sequence of SEQ ID NO: 5 and antisense sequence of SEQ ID NO: 58, siRNA-6 with sense sequence of SEQ ID NO: 6 and antisense sequence of SEQ ID NO: 59, siRNA-7 with sense sequence of SEQ ID NO: 7 and antisense sequence of SEQ ID NO: 60, siRNA-8 with sense sequence of SEQ ID NO: 8 and antisense sequence of SEQ ID NO: 61, siRNA-9 with sense sequence of SEQ ID NO: 9 and antisense sequence of SEQ ID NO: 62, siRNA-10 with sense sequence of SEQ ID NO: 10 and antisense sequence of SEQ ID NO: 63, siRNA-11 with sense sequence of SEQ ID NO: 11 and antisense sequence of SEQ ID NO: 64, siRNA-12 with sense sequence of SEQ ID NO: 12 and antisense sequence of SEQ ID NO: 65, siRNA-13 with sense sequence of SEQ ID NO: 13 and antisense sequence of SEQ ID NO: 66, siRNA-14 with sense sequence of SEQ ID NO: 14 and antisense sequence of SEQ ID NO: 67, siRNA-15 with sense sequence of SEQ ID NO: 15 and antisense sequence of SEQ ID NO: 68, siRNA-16 with sense sequence of SEQ ID NO: 16 and antisense sequence of SEQ ID NO: 69, siRNA-17 with sense sequence of SEQ ID NO: 17 and antisense sequence of SEQ ID NO: 70, siRNA-18 with sense sequence of SEQ ID NO: 18 and antisense sequence of SEQ ID NO: 71, siRNA-19 with sense sequence of SEQ ID NO: 19 and antisense sequence of SEQ ID NO: 72, siRNA-20 with sense sequence of SEQ ID NO: 20 and antisense sequence of SEQ ID NO: 73, siRNA-21 with sense sequence of SEQ ID NO: 21 and antisense sequence of SEQ ID NO: 74, siRNA-22 with sense sequence of SEQ ID NO: 22 and antisense sequence of SEQ ID NO: 75, siRNA-23 with sense sequence of SEQ ID NO: 23 and antisense sequence of SEQ ID NO: 76, siRNA-24 with sense sequence of SEQ ID NO: 24 and antisense sequence of SEQ ID NO: 77, siRNA-25 with sense sequence of SEQ ID NO: 25 and antisense sequence of SEQ ID NO: 78, siRNA-26 with sense sequence of SEQ ID NO: 26 and antisense sequence of SEQ ID NO: 79, siRNA-27 with sense sequence of SEQ ID NO: 27 and antisense sequence of SEQ ID NO: 80, siRNA-28 with sense sequence of SEQ ID NO: 28 and antisense sequence of SEQ ID NO: 81, siRNA-29 with sense sequence of SEQ ID NO: 29 and antisense sequence of SEQ ID NO: 82, siRNA-30 with sense sequence of SEQ ID NO: 30 and antisense sequence of SEQ ID NO: 83, siRNA-31 with sense sequence of SEQ ID NO: 31 and antisense sequence of SEQ ID NO: 84, siRNA-32 with sense sequence of SEQ ID NO: 32 and antisense sequence of SEQ ID NO: 85, siRNA-33 with sense sequence of SEQ ID NO: 33 and antisense sequence of SEQ ID NO: 86, siRNA-34 with sense sequence of SEQ ID NO: 34 and antisense sequence of SEQ ID NO: 87, siRNA-35 with sense sequence of SEQ ID NO: 35 and antisense sequence of SEQ ID NO: 88, siRNA-36 with sense sequence of SEQ ID NO: 36 and antisense sequence of SEQ ID NO: 89, siRNA-37 with sense sequence of SEQ ID NO: 37 and antisense sequence of SEQ ID NO: 90, siRNA-38 with sense sequence of SEQ ID NO: 38 and antisense sequence of SEQ ID NO: 91, siRNA-39 with sense sequence of SEQ ID NO: 39 and antisense sequence of SEQ ID NO: 92, siRNA-40 with sense sequence of SEQ ID NO: 40 and antisense sequence of SEQ ID NO: 93, siRNA-41 with sense sequence of SEQ ID NO: 41 and antisense sequence of SEQ ID NO: 94, siRNA-42 with sense sequence of SEQ ID NO: 42 and antisense sequence of SEQ ID NO: 95, siRNA-43 with sense sequence of SEQ ID NO: 43 and antisense sequence of SEQ ID NO: 96, siRNA-44 with sense sequence of SEQ ID NO: 44 and antisense sequence of SEQ ID NO: 97, siRNA-45 with sense sequence of SEQ ID NO: 45 and antisense sequence of SEQ ID NO: 98, siRNA-46 with sense sequence of SEQ ID NO: 46 and antisense sequence of SEQ ID NO: 99, siRNA-47 with sense sequence of SEQ ID NO: 47 and antisense sequence of SEQ ID NO: 100, siRNA-48 with sense sequence of SEQ ID NO: 48 and antisense sequence of SEQ ID NO: 101, siRNA-49 with sense sequence of SEQ ID NO: 49 and antisense sequence of SEQ ID NO: 102, siRNA-50 with sense sequence of SEQ ID NO: 50 and antisense sequence of SEQ ID NO: 103, siRNA-51 with sense sequence of SEQ ID NO: 51 and antisense sequence of SEQ ID NO: 104, siRNA-52 with sense sequence of SEQ ID NO: 52 and antisense sequence of SEQ ID NO: 105, and siRNA-53 with sense sequence of SEQ ID NO: 53 and antisense sequence of SEQ ID NO: 106, as shown in Table 1.siRNA-21 of NO: 74, siRNA-22 of which sense strand sequence is SEQ ID NO: 22 and antisense strand sequence is SEQ ID NO: 75, siRNA-23 of which sense strand sequence is SEQ ID NO: 23 and antisense strand sequence is SEQ ID NO: 76, siRNA-24 of which sense strand sequence is SEQ ID NO: 24 and antisense strand sequence is SEQ ID NO: 77, siRNA-25 of which sense strand sequence is SEQ ID NO: 25 and antisense strand sequence is SEQ ID NO: 78, siRNA-26 of which sense strand sequence is SEQ ID NO: 26 and antisense strand sequence is SEQ ID NO: 79, siRNA-27 of which sense strand sequence is SEQ ID NO: 27 and antisense strand sequence is SEQ ID NO: 80, siRNA-28 of which sense strand sequence is SEQ ID NO: 28 and antisense strand sequence is SEQ ID NO: 81, siRNA-29 of which sense strand sequence is SEQ ID NO: 29 and antisense strand sequence is SEQ ID NO: 82, siRNA-30 of which sense strand sequence is SEQ ID NO: 30 and antisense strand sequence is SEQ ID NO: 83, siRNA-31 of which sense strand sequence is SEQ ID NO: 31 and antisense strand sequence is SEQ ID NO: 84, siRNA-32 of which sense strand sequence is SEQ ID NO: 32 and antisense strand sequence is SEQ ID NO: 85, siRNA-33 of which sense strand sequence is SEQ ID NO: 33 and antisense strand sequence is SEQ ID NO: 86, siRNA-34 of which sense strand sequence is SEQ ID NO: 34 and antisense strand sequence is SEQ ID NO: 87, siRNA-35 of which sense strand sequence is SEQ ID NO: 35 and antisense strand sequence is SEQ ID NO: 88, siRNA-36 of which sense strand sequence is SEQ ID NO: 36 and antisense strand sequence is SEQ ID NO: 89, siRNA-37 of which sense strand sequence is SEQ ID NO: 37 and antisense strand sequence is SEQ ID NO: 90, siRNA-38 of which sense strand sequence is SEQ ID NO: 38 and antisense strand sequence is SEQ ID NO: 91, siRNA-39 of which sense strand sequence is SEQ ID NO: 39 and antisense strand sequence is SEQ ID NO: 92, siRNA-40 of which sense strand sequence is SEQ ID NO: 40 and antisense strand sequence is SEQ ID NO: 93, siRNA-41 of which sense strand sequence is SEQ ID NO: 41 and antisense strand sequence is SEQ ID NO: 94, siRNA-42 of which sense strand sequence is SEQ ID NO: 42 and antisense strand sequence is SEQ IDat least one of siRNA-42 of SEQ ID NO: 95, siRNA-43 having a sense strand sequence of SEQ ID NO: 43 and an antisense strand sequence of SEQ ID NO: 96, siRNA-44 having a sense strand sequence of SEQ ID NO: 44 and an antisense strand sequence of SEQ ID NO: 97, siRNA-45 having a sense strand sequence of SEQ ID NO: 45 and an antisense strand sequence of SEQ ID NO: 98, siRNA-46 having a sense strand sequence of SEQ ID NO: 46 and an antisense strand sequence of SEQ ID NO: 99, siRNA-47 having a sense strand sequence of SEQ ID NO: 47 and an antisense strand sequence of SEQ ID NO: 100, siRNA-48 having a sense strand sequence of SEQ ID NO: 48 and an antisense strand sequence of SEQ ID NO: 101, siRNA-49 having a sense strand sequence of SEQ ID NO: 49 and an antisense strand sequence of SEQ ID NO: 102, siRNA-50 having a sense strand sequence of SEQ ID NO: 50 and an antisense strand sequence of SEQ ID NO: 103, siRNA-51 having a sense strand sequence of SEQ ID NO: 51 and an antisense strand sequence of SEQ ID NO: 104, siRNA-52 having a sense strand sequence of SEQ ID NO: 52 and an antisense strand sequence of SEQ ID NO: 105, siRNA-53 having a sense strand sequence of SEQ ID NO: 53 and an antisense strand sequence of SEQ ID NO: 106.

[0049] In preferred embodiments of the application, the RNAi agent is siRNA-1 having a sense strand sequence of SEQ ID NO: 1 and an antisense strand sequence of SEQ ID NO: 54, siRNA-2 having a sense strand sequence of SEQ ID NO: 2 and an antisense strand sequence of SEQ ID NO: 55, siRNA-3 having a sense strand sequence of SEQ ID NO: 3 and an antisense strand sequence of SEQ ID NO: 56, siRNA-4 having a sense strand sequence of SEQ ID NO: 4 and an antisense strand sequence of SEQ ID NO: 57, siRNA-5 having a sense strand sequence of SEQ ID NO: 5 and an antisense strand sequence of SEQ ID NO: 58, siRNA-6 having a sense strand sequence of SEQ ID NO: 6 and an antisense strand sequence of SEQ ID NO: 59, siRNA-7 having a sense strand sequence of SEQ ID NO: 7 and an antisense strand sequence of SEQ ID NO: 60, siRNA-8 having a sense strand sequence of SEQ ID NO: 8 and an antisense strand sequence of SEQ ID NO: 61, siRNA-9 having a sense strand sequence of SEQ ID NO: 9 and an antisense strand sequence of SEQ ID NO: 62, siRNA-10 having a sense strand sequence of SEQ ID NO: 10 and an antisense strand sequence of SEQ ID NO: 63, siRNA-11 having a sense strand sequence of SEQ ID NO: 11 and an antisense strand sequence of SEQ ID NO: 64, siRNA-12 having a sense strand sequence of SEQ ID NO: 12 and an antisense strand sequence of SEQ ID NO: 65, siRNA-13 having a sense strand sequence of SEQ ID NO: 13 and an antisense strand sequence of SEQ ID NO: 66, siRNA-14 having a sense strand sequence of SEQ ID NO: 14 and an antisense strand sequence of SEQ ID NO: 67, siRNA-15 having a sense strand sequence of SEQ ID NO: 15 and an antisense strand sequence of SEQ ID NO: 68, siRNA-16 having a sense strand sequence of SEQ ID NO: 16 and an antisense strand sequence of SEQ ID NO: 69, siRNA-17 having a sense strand sequence of SEQ ID NO: 17 and an antisense strand sequence of SEQ ID NO: 70, siRNA-18 having a sense strand sequence of SEQ ID NO: 18 and an antisense strand sequence of SEQ ID NO: 71, siRNA-19 having a sense strand sequence of SEQ ID NO: 19 and an antisense strand sequence of SEQ ID NO: 72, siRNA-20 having a sense strand sequence of SEQ ID NO: 20 and an antisense strand sequence of SEQ ID NO: 73, siRNA-21 having a sense strand sequence of SEQ ID NO: 21 and an antisense strand sequence of SEQ IDsiRNA-21 of NO: 74, siRNA-22 of which sense strand sequence is SEQ ID NO: 22 and antisense strand sequence is SEQ ID NO: 75, siRNA-23 of which sense strand sequence is SEQ ID NO: 23 and antisense strand sequence is SEQ ID NO: 76, siRNA-24 of which sense strand sequence is SEQ ID NO: 24 and antisense strand sequence is SEQ ID NO: 77, siRNA-25 of which sense strand sequence is SEQ ID NO: 25 and antisense strand sequence is SEQ ID NO: 78, siRNA-26 of which sense strand sequence is SEQ ID NO: 26 and antisense strand sequence is SEQ ID NO: 79, siRNA-27 of which sense strand sequence is SEQ ID NO: 27 and antisense strand sequence is SEQ ID NO: 80, siRNA-28 of which sense strand sequence is SEQ ID NO: 28 and antisense strand sequence is SEQ ID NO: 81, siRNA-29 of which sense strand sequence is SEQ ID NO: 29 and antisense strand sequence is SEQ ID NO: 82, siRNA-30 of which sense strand sequence is SEQ ID NO: 30 and antisense strand sequence is SEQ ID NO: 83, siRNA-31 of which sense strand sequence is SEQ ID NO: 31 and antisense strand sequence is SEQ ID NO: 84, siRNA-32 of which sense strand sequence is SEQ ID NO: 32 and antisense strand sequence is SEQ ID NO: 85, siRNA-33 of which sense strand sequence is SEQ ID NO: 33 and antisense strand sequence is SEQ ID NO: 86, siRNA-34 of which sense strand sequence is SEQ ID NO: 34 and antisense strand sequence is SEQ ID NO: 87, siRNA-35 of which sense strand sequence is SEQ ID NO: 35 and antisense strand sequence is SEQ ID NO: 88, siRNA-36 of which sense strand sequence is SEQ ID NO: 36 and antisense strand sequence is SEQ ID NO: 89, siRNA-37 of which sense strand sequence is SEQ ID NO: 37 and antisense strand sequence is SEQ ID NO: 90, siRNA-38 of which sense strand sequence is SEQ ID NO: 38 and antisense strand sequence is SEQ ID NO: 91, siRNA-39 of which sense strand sequence is SEQ ID NO: 39 and antisense strand sequence is SEQ ID NO: 92, siRNA-40 of which sense strand sequence is SEQ ID NO: 40 and antisense strand sequence is SEQ ID NO: 93, siRNA-41 of which sense strand sequence is SEQ ID NO: 41 and antisense strand sequence is SEQ ID NO: 94, siRNA-42 of which sense strand sequence is SEQ ID NO: 42 and antisense strand sequence is SEQ IDone of siRNA-42 of SEQ ID NO: 95, siRNA-43 of which the sense strand sequence is SEQ ID NO: 43 and the antisense strand sequence is SEQ ID NO: 96, siRNA-44 of which the sense strand sequence is SEQ ID NO: 44 and the antisense strand sequence is SEQ ID NO: 97, siRNA-45 of which the sense strand sequence is SEQ ID NO: 45 and the antisense strand sequence is SEQ ID NO: 98, siRNA-46 of which the sense strand sequence is SEQ ID NO: 46 and the antisense strand sequence is SEQ ID NO: 99, siRNA-47 of which the sense strand sequence is SEQ ID NO: 47 and the antisense strand sequence is SEQ ID NO: 100, siRNA-48 of which the sense strand sequence is SEQ ID NO: 48 and the antisense strand sequence is SEQ ID NO: 101, siRNA-49 of which the sense strand sequence is SEQ ID NO: 49 and the antisense strand sequence is SEQ ID NO: 102, siRNA-50 of which the sense strand sequence is SEQ ID NO: 50 and the antisense strand sequence is SEQ ID NO: 103, siRNA-51 of which the sense strand sequence is SEQ ID NO: 51 and the antisense strand sequence is SEQ ID NO: 104, siRNA-52 of which the sense strand sequence is SEQ ID NO: 52 and the antisense strand sequence is SEQ ID NO: 105, siRNA-53 of which the sense strand sequence is SEQ ID NO: 53 and the antisense strand sequence is SEQ ID NO: 106.

[0050] In some preferred embodiments, the antisense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from the sequence set forth in any one of SEQ ID NOs: 54, 55, 57, 59, 62, 63, 67-69, 76, 80-82, 88-95, 98, 99, 101, 105, and the sense strand comprises a nucleotide sequence that is at least partially complementary (e.g., partially complementary, substantially complementary, or completely complementary) to the antisense strand. In some preferred embodiments, the sense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from the sequence set forth in any one of SEQ ID NOs: 1, 2, 4, 6, 9, 10, 14-16, 23, 27-29, 35-42, 45, 46, 48, 52. When comprising the sequences as above, the RNAi agent has a more superior inhibitory effect.

[0051] In some further preferred embodiments, the sense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from the sequence set forth in SEQ ID NO: 9, and the antisense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from the sequence set forth in SEQ ID NO: 62. In some preferred embodiments, the antisense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from the sequence set forth in any one of SEQ ID NOs: 54, 55, 57, 59, 62, 63, 67-69, 76, 80-82, 88-95, 98, 99, 101, 105, and the sense strand comprises a nucleotide sequence that is at least partially complementary (e.g., partially complementary, substantially complementary, or completely complementary) to the antisense strand. In some preferred embodiments, the sense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from the sequence set forth in any one of SEQ ID NOs: 1, 2, 4, 6, 9, 10, 14-16, 23, 27-29, 35-42, 45, 46, 48, 52. When comprising the sequences as above, the RNAi agent has a more superior inhibitory effect.

[0052] In some further preferred embodiments, the sense strand contains a nucleotide sequence that differs from the sequence set forth in SEQ ID NO: 10 by 0, 1 or 2 nucleotides, and the antisense strand contains a nucleotide sequence that differs from the sequence set forth in SEQ ID NO: 63 by 0, 1 or 2 nucleotides.

[0053] In some further preferred embodiments, the sense strand contains a nucleotide sequence that differs from the sequence set forth in SEQ ID NO: 38 by 0, 1 or 2 nucleotides, and the antisense strand contains a nucleotide sequence that differs from the sequence set forth in SEQ ID NO: 91 by 0, 1 or 2 nucleotides.

[0054] The inhibitory effect of the RNAi agent is further more preferably optimal when the sequences are as set forth in SEQ ID NO: 38 for the sense strand and SEQ ID NO: 91 for the antisense strand.

[0055] All of the nucleotide groups in the above RNAi agent can be unmodified or contain at least one modified nucleotide group, and the modification can be on any of the nucleotides.

[0056] In some embodiments, the sense strand and the antisense strand can be partially complementary, substantially complementary or fully complementary to each other.

[0057] In some embodiments, the sense strand or the antisense strand in the RNAi agent has a sequence identity of less than 100% or differs from the corresponding sequence mentioned in the present application by more than 1 nucleotide, and still has an inhibitory effect on C3 complement similar to (e.g., still has an efficacy equivalent to 80-120%, 85-115% or 90-110% of the corresponding sequence) or equivalent to (e.g., still has an efficacy equivalent to 95-105% of the corresponding sequence) the corresponding sequence. For example, the two bases at the 3' end of the antisense strand (e.g., the sequence set forth in any one of SEQ ID NO: 54-106) are replaced by UU, AA, CU, UC, AG, CC, GG or UG, etc., or any combination of two nucleic acids. Such nucleic acid sequences also fall within the scope of the present application.

[0058] In some preferred embodiments, the RNAi agent inhibits C3 complement with an efficiency of no less than 50% (e.g., 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%).

[0059] The present application also provides a target gene site of the nucleic acid as described above. In some embodiments, the target gene site is as labeled in any one of the first column of Table 1.

[0060] Table 1

[0061] Note: The first column refers to the position of the first base of the target gene in the human C3 complement mRNA sequence, and so on; the numbers in the third and fifth columns represent sequence numbers, for example, “1” represents SEQ ID NO: 1.

[0062] In some embodiments, the reference sequence of the target gene is the coding sequence of human C3 complement mRNA, NM_000064.4.

[0063] In some embodiments, the sense strand of the RNAi agent has at least 80% (e.g., 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) sequence identity to an equal-length sequence in the nucleotides at positions 200-400, 400-600, 600-900, 1200-1400, 1300-1500, 2500-2600, 2800-3000, 3700-3900, or 4000-4700 of NM_000064.4.

[0064] In some embodiments, the RNAi agent targets any position within the region of nucleotides at positions 200-400, 400-600, 600-900, 1200-1400, 1300-1500, 2500-2600, 2800-3000, 3700-3900, or 4000-4700 of NM_000064.4.

[0065] In some embodiments, the RNAi agent targets any position within the region of nucleotides 200-300, 400-500, 650-750, 700-850, 1250-1350, 1350-1450, 2500-2600, 2800-2950, 3750-3850, 4100-4300, 4400-4550, or 4550-4650 of NM_000064.4.

[0066] In some embodiments, the RNAi agent targets any position within the region of nucleotides 760-810 of NM_000064.4.

[0067] In some embodiments, the RNAi agent targets any position within the region of nucleotides 2840-2900 of NM_000064.4.

[0068] In some embodiments, the RNAi agent targets any position within the region of nucleotides 3770-3820 of NM_000064.4.

[0069] In some embodiments, the RNAi agent targets any position within the region of nucleotides 4110-4160 of NM_000064.4.

[0070] In some embodiments, the RNAi agent targets any position within the region of nucleotides 4590-4640 of NM_000064.4.

[0071] The above technical solutions mentioned in the present application regarding naked sequence (i.e. unmodified sequence) have effect advantages that do not depend on the selection of modification method or targeting vector. The following introduces the modification schemes that can be applied and further preferred modification schemes:

[0072] In some embodiments, the RNAi agent contains nucleotide groups as the basic structural units, which contain a phosphate group, a ribose group and a base, preferably, the RNAi agent contains at least one modified nucleotide group. The RNAi agent containing the modified group has an inhibition efficiency of no less than 50% (such as 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%) on C3 complement.

[0073] In some embodiments, the modified nucleotide group is a nucleotide group with a modified phosphate group and / or ribose group. The modified site can be at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 nucleotides in the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th nucleotide of the sense strand and / or the antisense strand.

[0074] In some embodiments, the modification of the phosphate group refers to the modification of the oxygen in the phosphate group, including phosphorthioate and boranophosphate modification, etc. The oxygen in the phosphate group is replaced by sulfur, borane, amine group, alkyl or alkoxy group, respectively, as shown in the following formula. These modifications can stabilize the structure of the nucleic acid, maintain high specificity and high affinity of base pairing.

[0075] In the above structural formula, BASE represents a base A, U, C, G, or T. X can be oxygen (O) or sulfur (S). R can be the same or different in the above structure, such as hydrogen (H), fluorine (F), methoxy (OME), or methoxyethyl (MOE), hydroxyl, allyl, ethylamine, propargyl, amino, cyanoethyl, acetyl, etc., and R' and R" can each independently be hydrogen (H), methyl (CH3), ethyl (CH2CH3), propyl (CH2CH2CH3), isopropyl (CH(CH3)2), allyl, propargyl, acyloxybenzyl, acyloxyethyl.

[0076] In some embodiments, the modification of the ribose group refers to the modification of the 2'-hydroxyl group (2'-OH) in the ribose group. After introducing certain substituents such as methoxy or fluorine at the 2'-hydroxyl position of the ribose group, the nucleic acid is less susceptible to cleavage by ribonuclease, thereby increasing the stability of the nucleic acid and making the nucleic acid have stronger resistance to nucleic acid enzyme hydrolysis. The modification of the 2'-hydroxyl group in the pentose of the nucleotide includes 2'-fluoromodification (such as 2'-arabino-fluoro modification), 2'-methoxy modification (2'-OME), 2'-methoxyethyl modification (2'-MOE), 2'-2,4-dinitrophenol modification (2'-DNP modification), 2',4'-constrained ethyl modification, 2'-amino modification, 2'-deoxy modification, BNA, acyclic nucleic acid modification, mispaired nucleic acid modification, L-type nucleic acid modification, etc. BNA (internally bridged nucleic acid) refers to a constrained or inaccessible nucleotide. BNA can contain a five-membered ring, a six-membered ring, or a seven-membered ring with a "fixed" C 3'-endo sugar contraction bridge structure. The bridge is usually incorporated into the 2'-, 4'-position of the ribose ring to provide a 2',4'-BNA nucleotide, such as a locked ethyl modification (LNA), a constrained ethyl modification (ENA), and an ethyl locked nucleic acid modification (cET BNA). Acyclic nucleic acid is a nucleotide whose sugar ring is opened to form a nucleotide, such as an unlocked nucleic acid (UNA) nucleotide and a glycerol nucleic acid (GNA) nucleotide. Mispaired nucleic acid modification refers to the replacement of 3',5'-phosphodiester bonds with 2',5'-phosphodiester bonds. L-type nucleic acid modification refers to the replacement of naturally occurring D-type nucleic acid with its mirror image stereoisomer L-type nucleic acid.

[0077] wherein BASE represents a base A, U, C, G or T. R in the above structure can be the same or different, such as: hydrogen (H), fluorine (F), methoxy (OME) or methoxyethyl (MOE), hydroxyl, allyl, ethylamine, propargyl, cyanoethyl, acetyl, etc.

[0078] In some embodiments, the nucleotide group containing a uracil base or a cytosine base in the sense strand of the RNAi agent is a nucleotide group with a modified ribose group, i.e., the 2’-OH of the ribose group in the nucleotide group containing a uracil base or a cytosine base in the sense strand of the RNAi agent is substituted with a methoxy group or a fluorine. More preferably, the 3’ end of the sense strand and the antisense strand of the RNAi agent can both be linked with dTdT; or the 3’ end of the antisense strand of the RNAi agent can be linked with AA or UU or any combination of two nucleic acids (which can be but is not limited to CC, GG or UG) to make the sequence have a specific inducement for mRNA degradation. The RNAi agent with the above modification exhibits more excellent inhibitory effect in vivo, and the above modification can further reduce the immunogenicity of the RNAi agent of the present application in vivo.

[0079] The RNAi agent of the present application can also include a modification of linking a monophosphate nucleoside at the 5’ end of the antisense strand. Since the 5’-monophosphate at the terminal end of the guide strand of siRNA is important for the recognition of RISC. The phosphorylation of the 5’-hydroxyl group plays a certain role in whether the siRNA can be effectively loaded onto Ago2 inside the cell. The 5’-monophosphate at the terminal end of the guide strand of siRNA has H-bond interaction with Argonaute-2 (Ago2), thereby ensuring accurate targeting and precise cleavage of the mRNA target. Commonly used derivatives of 5’-monophosphate nucleosides include the following several kinds. Such derivatives of monophosphate nucleosides have been shown to have certain stability in biological metabolic media and have a certain effect on promoting the loading of siRNA guide strand to Ago2 inside the cell (Nucleic Acids Research, 2015, 43, 2993-3011). According to the RNAi agent of the present application, preferably, trans-vinyl phosphate (VP) is used as the first choice, and derivatives of monophosphate nucleosides other than those described above can also be included.

[0080] In the above structure, BASE represents a base A, U, C, G or T. R in the above structure can be the same or different, such as: hydrogen (H), fluorine (F), methoxy (OME) or methoxyethyl (MOE), hydroxyl, allyl, ethylamine, propargyl, cyanoethyl, acetyl, etc.

[0081] In the present application, The term "linked to" has the same meaning as "attached to" and means that a chemical element X is attached to any one or more groups.

[0082] In some embodiments, at least one nucleotide in the RNAi agent is a modified nucleotide or comprises a modified internucleotide linkage.

[0083] In some embodiments, the modified nucleotide is preferably selected from one or more of a 2'-0-methyl nucleotide, a 2'-fluoro nucleotide, a 2'-deoxy nucleotide, a 2',3'-seco nucleotide mimic, a locked nucleotide, a 2'-F-arabino nucleotide, a 2'-methoxyethyl nucleotide, an abasic nucleotide, a ribo-ol, an inverted nucleotide, an inverted 2'-0-methyl nucleotide, an inverted 2'-deoxy nucleotide, a 2'-amino modified nucleotide, a 2'-alkyl modified nucleotide, a morpholino, a peptide nucleic acid (PNA), a glycerol nucleic acid (GNA), a triazolyl-cyclo DNA (tcDNA), a nucleotide comprising a non-natural base, a nucleotide comprising a vinyl phosphonate, a nucleotide comprising a cyclopropyl phosphonate, and a 3'-0-methyl nucleotide. In some embodiments, the modified nucleotide is further preferably selected from one or both of a 2'-0-methyl nucleotide and a 2'-fluoro nucleotide.

[0084] In some embodiments, the modified internucleotide linkage is preferably selected from one or more of a phosphorothioate internucleotide linkage and a methylphosphonate internucleotide linkage. In some embodiments, the modified internucleotide linkage is further preferably selected from one or more of a phosphorothioate monoester internucleotide linkage and a phosphorothioate diester internucleotide linkage.

[0085] In some embodiments, the antisense strand contains a phosphorothioate internucleotide linkage between the last 2, 3, or 4 nucleotides of the 5' end and / or the 3' end of the antisense strand, and contains 4, 5, 6, 7, or 8 2'-fluoro nucleotides in the antisense strand, with the remaining nucleotides being 2'-0-methyl nucleotides.

[0086] In some alternative embodiments, the antisense strand contains a phosphorothioate internucleotide linkage between the last 3 nucleotides of the 5' end and the 3' end of the antisense strand, and contains at least 1 (preferably 2, 3, 4, 5, or 6) 2'-fluoro nucleotides at nucleotides numbered 2, 4, 6, 8, 10, 12, 14, 16, 18 from the 5' end, with the remaining nucleotides being 2'-0-methyl nucleotides.

[0087] In some embodiments, the antisense strand comprises 5'-nNfnNfnNfnnnnnnnNfnNfnnnnnnn-3', 5'-nNfnnnNfnnnnnnnNfnNfnnnnnnn-3', 5'-nNfnNfnnnnnnnnnNfnNfnnnnnnn-3', 5'-nNfnNfnNfnNfnnnnnNfnNfnnnnnnn-3', 5'-nNfnNfnNfnnnNfnnnNfnNfnnnnnnn-3', 5'-nNfnNfnNfnnnnnNfnNfnNfnnnnnnn-3', or 5'-nNfnNfnNfnnnnnnnNfnNfnNfnnnnn-3'; wherein Nf represents a 2'-fluoro nucleotide, and n represents a 2'-O-methyl nucleotide. Compared to known modification, the above antisense strand modification further facilitates the inhibition of C3 complement by the RNAi agent.

[0088] In some preferred embodiments, the antisense strand comprises 5'-nNfnNfnNfnnnnnnnNfnNfnnnnnnn-3'. Compared to other modification, the above antisense strand modification significantly facilitates the inhibition of C3 complement.

[0089] In some embodiments, the last 2, 3, or 4 nucleotides at the 5' end and / or the 3' end of the sense strand comprise phosphorothioate internucleotide linkages, and the sense strand comprises 2, 3, 4, 5, or 6 2'-fluoro nucleotides, and the remaining nucleotides are 2'-O-methyl nucleotides.

[0090] In some embodiments, the sense strand comprises at least 1 (preferably 2, 3, or 4) of the 2'-fluoro nucleotides at the 7th, 9th, 10th, and 11th nucleotides counting from the 5' end, and the remaining nucleotides are 2'-O-methyl nucleotides.

[0091] In some embodiments, the sense strand comprises 5'-nnnnnnNfnNfNfNfnnnnnnnnnn-3'; wherein Nf represents a 2'-fluoro nucleotide, and n represents a 2'-O-methyl nucleotide. Compared to known modification, the above sense strand modification further facilitates the inhibition of C3 complement by the RNAi agent.

[0092] In some embodiments, the last 2-4 (e.g., 2, 3, or 4) nucleotides at the 5' end and / or the 3' end of the antisense strand comprise phosphorothioate internucleotide linkages, and the last 2-4 (e.g., 2, 3, or 4) nucleotides at the 5' end and / or the 3' end of the sense strand comprise phosphorothioate internucleotide linkages.

[0093] In some embodiments, the 5' terminal and 3' terminal last 3 nucleotides of the antisense strand contain phosphorothioate internucleotide linkages, and the 5' terminal last 3 nucleotides of the sense strand contain phosphorothioate internucleotide linkages.

[0094] In some embodiments, the antisense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from any of the antisense strands set forth in Table 2 or Table 3.

[0095] In some embodiments, the sense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from any of the sense strands set forth in Table 2 or Table 3.

[0096] In some embodiments, the RNAi agent comprises a duplex set forth in any of Table 2 or Table 3.

[0097] In some preferred embodiments, the RNAi agent comprises a duplex selected from any of PAT-0001, PAT-0002, PAT-0004, PAT-0006, PAT-0009, PAT-0009.1, PAT-0009.2, PAT-0010, PAT-0010.1, PAT-0010.2, PAT-0014, PAT-0015, PAT-0016, PAT-0023, PAT-0027, PAT-0028, PAT-0029, PAT-0035, PAT-0036, PAT-0037, PAT-0038, PAT-0038.1, PAT-0038.2, PAT-0038.3, PAT-0038.4, PAT-0038.5, PAT-0039, PAT-0040, PAT-0041, PAT-0042, PAT-0045, PAT-0046, PAT-0048, PAT-0052. RNAi agents comprising the above-mentioned duplexes are able to achieve better inhibitory effect on C3 complement in cell experiments.

[0098] In some further preferred embodiments, the RNAi agent comprises a duplex selected from any of PAT-0009, PAT-0010, PAT-0038. RNAi agents comprising the above-mentioned duplexes are able to achieve better inhibitory effect on C3 complement in vitro and in vivo.

[0099] In some further preferred embodiments, the RNAi agent comprises a duplex set forth in PAT-0038. RNAi agents comprising the above-mentioned duplex are able to achieve the best inhibitory effect on C3 complement in vitro and in vivo.

[0100] Table 2

[0101] Table 3

[0102] In the modified sequences of the present application, the nucleotides represented by lower case letters represent 2'-O-methyl nucleotides; f represents that the nucleotide adjacent to the left is a 2'-fluoro nucleotide; s represents that the two nucleotides adjacent to the left and right are connected by a phosphorothioate internucleotide linkage.

[0103] The RNAi agent according to the present application can be obtained by methods conventional in the art, for example by solid phase synthesis and liquid phase synthesis, the former of which is already commercially available and thus can be obtained by purchase. The modified nucleotide groups can be introduced by using nucleotide monomers having the corresponding modifications.

[0104] Based on the RNAi agent (siRNA) synthesized as above, the present application can further construct an shRNA expression plasmid having the same or similar function as the above-mentioned RNAi agent, and the method for constructing the expression plasmid is well known to those skilled in the art and thus will not be described here.

[0105] In some embodiments, the RNAi agent further comprises at least one ligand attached to one or more of the 5' end of the antisense strand, the 3' end of the antisense strand, the 5' end of the sense strand, and the 3' end of the sense strand.

[0106] In combination with the common knowledge in the art, the sense strand and the antisense strand of the present application have a better inhibitory effect when applied to different targeted drug delivery systems. In other words, the effect advantage of the naked sequence and the modified sequence in the present application does not depend on the selection of the targeting carrier. In order to further improve the bioavailability and therapeutic effect of siRNA, the targeting drug delivery system is also optimized in the present application, and the following technical solutions are obtained.

[0107] In some embodiments, the ligand is attached to the 5' end or the 3' end of the sense strand of the nucleic acid. In some embodiments, the ligand is attached to the 5' end of the sense strand of the nucleic acid. In some embodiments, the ligand is attached to the 3' end of the sense strand of the nucleic acid.

[0108] In some embodiments, the ligand is attached to the 5' end or the 3' end of the antisense strand of the nucleic acid. In some embodiments, the ligand is attached to the 5' end of the antisense strand of the nucleic acid. In some embodiments, the ligand is attached to the 3' end of the antisense strand of the nucleic acid.

[0109] In some embodiments, the ligand is linked to the 3' end of the sense strand in the nucleic acid.

[0110] In some embodiments, the ligand is a GalNAc derivative; more preferably, the ligand is one or more GalNAc derivatives linked by a single-stranded, double-stranded or triple-stranded branched linker.

[0111] In some embodiments, the RNAi agent comprises a compound having the following structure I:

[0112] In structure I, Nu represents a nucleic acid consisting of the sense strand and the antisense strand. The targeted drug delivery system utilizes the structural feature on its left side to improve the cell penetration ability of the nucleic acid drug (Nu), enhance its stability in cells, and has a simple preparation process and strong practicability.

[0113] Cell

[0114] The present application also provides an ex vivo cell comprising the RNAi agent.

[0115] In some embodiments, the cell can be used for gene function research, disease model research, drug screening, etc.

[0116] In some embodiments, the cell does not develop into an animal individual. In some specific embodiments, the cell can be a microbial cell or an animal cell, but the animal cell is not an embryonic stem cell of an animal and cells at various stages of formation and development thereof (e.g., germ cells, zygote cells, etc.).

[0117] Pharmaceutical composition

[0118] The present application also provides a pharmaceutical composition comprising the RNAi agent.

[0119] The pharmaceutical composition can be prepared by a conventional method from the RNAi agent and the pharmaceutically acceptable carrier. For example, the pharmaceutical composition can be an injection solution. The injection solution can be used for subcutaneous, intramuscular or intravenous injection.

[0120] According to the pharmaceutical composition of the present application, the amount of the RNAi agent and the pharmaceutically acceptable carrier is not particularly limited, and generally, the content of the pharmaceutically acceptable carrier can be 1-100000 parts by weight (e.g., 1 part by weight, 5 parts by weight, 10 parts by weight, 50 parts by weight, 100 parts by weight, 500 parts by weight, 1000 parts by weight, 5000 parts by weight, 10000 parts by weight, 50000 parts by weight, 100000 parts by weight or any value between any two of the above values) relative to 1 part by weight of the RNAi agent.

[0121] According to the pharmaceutical composition of the present application, the pharmaceutically acceptable carrier can be various carriers conventionally used in the art, for example, can include at least one of a pH buffer, a protective agent, and an osmotic pressure adjusting agent. The pH buffer can be a tris-hydroxymethyl aminomethane hydrochloride buffer having a pH of 7.5-8.5 and / or a phosphate buffer having a pH of 5.5-8.5, preferably a phosphate buffer having a pH of 5.5-8.5. The protective agent can be at least one of myo-inositol, sorbitol, and sucrose. The content of the protective agent can be 0.01-30% by weight (e.g., 0.01%, 0.05%, 0.1%, 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, or any value between any two of the above values) based on the total weight of the pharmaceutical composition. The osmotic pressure adjusting agent can be sodium chloride and / or potassium chloride. The content of the osmotic pressure adjusting agent can be such that the osmotic pressure of the pharmaceutical composition is 200-700 milliosmoles / kilogram. The content of the osmotic pressure adjusting agent can be determined by one skilled in the art according to the desired osmotic pressure.

[0122] According to a preferred embodiment of the present application, the pharmaceutically acceptable carrier is a liposome. The liposome can be any liposome capable of encapsulating a nucleic acid, and can have a diameter of 25-1000 nm, and can include, but is not limited to, cholesterol and analogs or derivatives thereof.

[0123] The dosage of the pharmaceutical composition of the present application can be a conventional dosage in the art, and can be determined according to various parameters, in particular according to the age, weight, and sex of the subject. For example, for a female mouse of 3-4 months of age and weighing 25-30 g, the dosage of the pharmaceutical composition can be 0.01-100 mg / kg of body weight, preferably 1-10 mg / kg of body weight, in terms of the amount of the RNAi agent in the pharmaceutical composition.

[0124] Methods and uses

[0125] The present application also provides a method of inhibiting the expression of C3 complement in a cell, the method comprising: contacting the cell with the RNAi agent or the pharmaceutical composition to inhibit the expression of C3 complement in the cell.

[0126] In some embodiments, the cell is in a subject, e.g., a human subject, e.g., a subject having a C3 complement-related disease, or a subject at risk of developing a C3 complement-related disease.

[0127] In some embodiments, the cell is in vitro. The method is based on research purposes, or for constructing an animal model.

[0128] In some embodiments, contacting the cell with the nucleic acid inhibits expression of C3 complement by at least 50%, 60%, 70%, 80%, 90%, 95% (e.g., as compared to the level of expression of C3 complement prior to the cell first contacting the nucleic acid; e.g., prior to administering a first dose of the nucleic acid to the subject). In certain embodiments, inhibiting expression of C3 complement reduces the level of C3 complement protein in a serum sample of the subject by at least 50%, 60%, 70%, 80%, 90%, or 95%, e.g., as compared to the level of expression of C3 complement prior to the cell first contacting the nucleic acid.

[0129] The present application also provides use of the RNAi agent or the pharmaceutical composition for treating and / or preventing a disease associated with C3 complement.

[0130] The present application also provides use of the RNAi agent or the pharmaceutical composition for the manufacture of a medicament for treating and / or preventing a disease associated with C3 complement.

[0131] In some embodiments, the disease is: (i) a disease associated with C3 complement enhancement or elevation; or (ii) a disease that would benefit from a reduction in C3 complement expression.

[0132] In some embodiments, the disease is selected from one or more of the following diseases: paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis, hereditary angioedema (HAE), multiple sclerosis, systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), Sjogren’s syndrome, membranoproliferative glomerulonephritis (MPGN), IgA nephropathy (lgAN), primary membranous nephropathy (primary MN), C3 glomerulopathy (C3G), acute tubular necrosis (ATN), focal segmental glomerulosclerosis (FSGS), diabetic nephropathy (DN), neuromyelitis optica (NMO), age-related macular degeneration (AMD), Guillain-Barre syndrome, multifocal motor neuropathy, and generalized myasthenia gravis (GMG).

[0133] In the present application, the subject can be a mammal, including a primate (e.g., a human, a non-human primate such as a monkey and a chimpanzee), a non-primate (e.g., a cow, a pig, a horse, a goat, a rabbit, a sheep, a hamster, a guinea pig, a cat, a dog, a rat, or a mouse), or a bird. In some embodiments, the subject is preferably a primate, more preferably a human.

[0134] In some embodiments, administration can be by a variety of routes, depending on whether local or systemic treatment is desired. Administration can be in amounts as described above, and need not be repeated more than once per day, or even once per week, depending on the severity of the disease and the potency and half-life of the RNAi agent.

[0135] In some embodiments, administration can be local (e.g., via a transdermal patch), pulmonary, e.g., by inhalation or insufflation of powders or aerosols, including by nebulizers; intratracheal, intranasal, epidermal and transdermal, oral or parenteral. Parenteral administration includes subcutaneous, intravenous, intramuscular, intraarterial, intraperitoneal or intramuscular injection or infusion; subepidermal, e.g., by means of a transdermal patch; or intracranial, e.g., by means of intracerei or, intrathecal or intraventricular administration.

[0136] In some embodiments, the RNAi agent or the pharmaceutical composition is administered to the subject by subcutaneous administration, intravenous administration, and / or intramuscular administration.

[0137] Examples

[0138] Embodiments of the application will now be described in detail with reference to the following examples. It is understood that these examples are for illustrative purposes only and are not meant to limit the scope of the application. Unless otherwise indicated, the experimental procedures in the following examples follow the guidelines set forth in the present application, and can also follow the procedures described in standard laboratory manuals or under conventional conditions, or can follow other procedures known in the art, or as suggested by the manufacturer.

[0139] In the following specific examples, the measurement parameters of the raw materials components, such as the amount, can have slight deviations within the range of the weighing accuracy, unless otherwise specified. The parameters of temperature and time are allowed to have acceptable deviations caused by the instrument testing accuracy or the operation accuracy.

[0140] Example 1

[0141] In a 96-well cell culture dish, 0.5 ml of cell culture medium (DMEM, 10% calf serum, 1% penicillin + streptomycin solution) containing 10 4Hep3B (Procell, Cat# CL-0102) cells were cultured overnight at 37 °C in a 5% CO2 incubator. RNAiMAX (1.5 μL / well) and small interfering nucleic acids (siRNAs) in Table 2 were added to the cells in the wells of a 96-well plate to a final concentration of 1 nM or 10 nM and incubated for 48 hours at 37 °C in a 5% CO2 incubator. To extract RNA, the cell culture supernatant was aspirated, washed with PBS, and 50 μL of prepared lysis solution (as recommended in the Cells-to-CT kit (Thermo Fisher Scientific, Cat# 4391851c)) was added and mixed, and after 10 minutes of standing, 2.5 μL of Stop solution was added and incubated for 2 minutes. RT-PCR was performed according to the recommendations of the High Capacity cDNA Reverse Transcription Kits (Thermo Fisher, Cat# 4368814), and 10 μL of the lysed solution was included in each reaction. Gene expression was quantified by real-time fluorescent PCR, and the TaqMan probe for human C3 was Hs00163811_m1, and the probe for the reference gene (human HPRT1) was Hs02800695_m1 (Thermo Fisher Scientific, Waltham, MA, USA). The PCR conditions were 1 cycle of 95 °C for 20 seconds, 40 cycles of 95 °C for 1 second and 60 °C for 20 seconds, and the real-time fluorescent PCR instrument was a QuantStudio 5 Real-Time PCR System (Thermo Fisher). C3 gene expression was calculated as 2^-AACt, and human HPRT1 gene expression was used as a reference. C3 gene expression was expressed as a percentage of the relative value of the control group with only RNAiMAX. The results are shown in Table 4 below. TM 6 Pro real-time fluorescent quantitative PCR system (Thermo Fisher). C3 gene expression was calculated as 2^-AACt, and human HPRT1 gene expression was used as a reference. C3 gene expression was expressed as a percentage of the relative value of the control group with only RNAiMAX. The results are shown in Table 4 below.

[0142] Table 4 Knockdown effect of siRNAs on C3 mRNA expression in Hep3B cells

[0143] Example 2

[0144] The siRNAs preferred in Example 1 were further diluted and further verified in Hep3B cells according to the method described above. The inhibitory effect of the siRNAs on the expression of the C3 gene was expressed as a percentage of the relative value of the control group with only RNAiMAX. The results are shown in Table 5 below.

[0145] Table 5

[0146] Example 3

[0147] As shown in Table 6, the siRNA preferred in Example 2 was coupled with Tri-GalNAc (the structure of the compound is shown in Formula I) at the 3' end of the sense strand, and was injected subcutaneously into human C3 transgenic mice (Shanghai Southern Model Organism Science and Technology Co., Ltd.) at a dose of 3 mg / kg. Blood was collected on day 14, and the human C3 protein level in the plasma was measured using an ELISA kit (Abeam, ab108823). PBS was used as a control group in this experiment. The results are shown in Figure 1.

[0148] Table 6

[0149] Example 4

[0150] In a 96-well cell culture dish, 0.5 mL of cell culture medium (DMEM, 10% calf serum, 1% penicillin + streptomycin solution) containing 10 4 Hep3B (Procell, Cat#CL-0102) cells were cultured overnight in a cell incubator at 37°C, 5% CO2. RNAiMAX (1.5 μL / well) and small interfering nucleic acid (siRNA) in Table 7 were added to the cell culture medium to a final concentration of 0.000038 nM, 0.000305 nM, 0.002441 nM, 0.019531 nM, 0.156250 nM, 1.25 nM or 10 nM per well, and the cells were further cultured for 48 hours in a cell incubator at 37°C, 5% CO2. To extract RNA, the cell culture supernatant was aspirated, washed with PBS, and 50 μL of prepared lysis solution (as recommended by the Cells-to-CT kit (Thermo Fisher Scientific, Cat#4391851c)) was added and mixed well. After standing for 10 min, 2.5 μL of Stop solution was added and terminated for 2 min. RT-PCR was performed according to the recommendations of the High Capacity cDNA Reverse Transcription Kits (Thermo Fisher, Cat#4368814), and 10 μL of the lysed solution was included in each reaction. Real-time fluorescent PCR was used to quantify gene expression, and the TaqMan probe for human C3 was Hs00163811_m1, and the probe for the internal reference gene (human HPRT1) was Hs02800695_m1 (Thermo Fisher Scientific, Waltham, MA, USA). The PCR conditions were 95°C for 20 seconds for 1 cycle, 95°C for 1 second and 60°C for 20 seconds for 40 cycles, and the real-time fluorescent PCR instrument was a QuantStudio 7 Flex Real-Time PCR System (Thermo Fisher Scientific, Waltham, MA, USA). TM6 Pro Real-Time PCR System (Thermo Fisher). C3 gene expression was calculated as 2^-AACt, with human HPRT1 gene expression as the internal control. C3 gene expression was expressed as a percentage of the relative value of the control group with only RNAiMAX, and the IC50 value was calculated by Prism GraphPad. The results are shown in Table 8.

[0151] Table 7

[0152] Table 8

[0153] Example 5

[0154] PAT-0038 also showed long-lasting inhibitory effect in human C3 transgenic mice. As shown in Figure 2, a single 3 mg / kg PAT-0038 T subcutaneous injection was able to sustain C3 expression inhibition by about 90% for at least one month.

[0155] The above described embodiments only express some preferred embodiments of the present application, which are described in more detail and in more detail, but cannot be understood as limiting the scope of the patent. It should be noted that for those skilled in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which are within the scope of the present application.

Claims

1. An RNAi agent, comprising: an antisense strand comprising at least 15 contiguous nucleotides that differ by no more than 3 nucleotides from any of the at least 15 contiguous nucleotides of any of the sequences set forth in SEQ ID NOs: 54-106; and a sense strand comprising a nucleotide sequence that is at least partially complementary to the antisense strand.

2. The RNAi agent of claim 1, wherein, the sense strand comprises at least 15 contiguous nucleotides that differ by no more than 3 nucleotides from any of the at least 15 contiguous nucleotides of any of the sequences set forth in SEQ ID NOs: 1-53.

3. The RNAi agent of claim 1, wherein, the antisense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from the sequence set forth in any of SEQ ID NOs: 54, 55, 57, 59, 62, 63, 67-69, 76, 80-82, 88-95, 98, 99, 101, 105, and the sense strand comprises a nucleotide sequence that is at least partially complementary to the antisense strand; optionally, the sense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from the sequence set forth in any of SEQ ID NOs: 1, 2, 4, 6, 9, 10, 14-16, 23, 27-29, 35-42, 45, 46, 48, 52.

4. The RNAi agent of claim 1, wherein, the sense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from the sequence set forth in SEQ ID NO: 9, and the antisense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from the sequence set forth in SEQ ID NO: 62; or, the sense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from the sequence set forth in SEQ ID NO: 10, and the antisense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from the sequence set forth in SEQ ID NO: 63; or, the sense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from the sequence set forth in SEQ ID NO: 38, and the antisense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from the sequence set forth in SEQ ID NO:

91.

5. The RNAi agent according to any one of claims 1-4, wherein, at least one nucleotide in the RNAi agent is a modified nucleotide or comprises a modified intemucleotide linkage; the modified nucleotides are preferably one or more of 2'-O-methyl nucleotides, 2'-fluoro nucleotides, 2'-deoxy nucleotides, 2',3'-seco nucleotide mimics, locked nucleotides, 2'-F-arabinose nucleotides, 2'-methoxyethyl nucleotides, abasic nucleotides, ribothymidines, inverted nucleotides, inverted 2'-O-methyl nucleotides, inverted 2'-deoxy nucleotides, 2'-amino modified nucleotides, 2'-alkyl modified nucleotides, morpholino nucleotides, peptide nucleic acids, glycerol nucleic acids, triazolyl DNA, nucleotides comprising non-natural bases, nucleotides comprising vinyl phosphonates, nucleotides comprising cyclopropyl phosphonates, and 3'-O-methyl nucleotides; the modified nucleotides are further preferably one or more of 2'-O-methyl nucleotides and 2'-fluoro nucleotides; the modified internucleotide linkages are preferably one or more of phosphorothioate internucleotide linkages and methylphosphonate internucleotide linkages; the modified internucleotide linkages are further preferably one or more of phosphorothioate monoester internucleotide linkages, phosphorothioate diester internucleotide linkages.

6. The RNAi agent of claim 5, wherein, the antisense strand comprises 5'-nNfnNfnNfnnnnnnnNfnNfnnnnnnn-3', 5'-nNfnnnNfnnnnnnnNfnNfnnnnnnn-3', 5'-nNfnNfnnnnnnnnnNfnNfnnnnnnn-3', 5'-nNfnNfnNfnNfnnnnnNfnNfnnnnnnn-3', 5'-nNfnNfnNfnnnNfnnnNfnNfnnnnnnn-3', 5'-nNfnNfnNfnnnnnNfnNfnNfnnnnnnn-3', or 5'-nNfnNfnNfnnnnnnnNfnNfnNfnnnnn-3'; wherein Nf represents a 2'-fluoro nucleotide and n represents a 2'-O-methyl nucleotide.

7. The RNAi agent of claim 5 or 6, wherein, the sense strand comprises 5'-nnnnnnNfnNfNfNfnnnnnnnnnn-3'; wherein Nf represents a 2'-fluoro nucleotide and n represents a 2'-O-methyl nucleotide.

8. The RNAi agent according to any one of claims 5-7, wherein, the antisense strand comprises a phosphorothioate internucleotide linkage between the last 2 to 4 nucleotides of the 5' end and / or the 3' end.

9. The RNAi agent according to any one of claims 5-8, wherein, the sense strand comprises a phosphorothioate internucleotide linkage between the last 2 to 4 nucleotides of the 5' end and / or the 3' end.

10. The RNAi agent of claim 1, wherein, the antisense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from any one of the antisense strands set forth in Table 2 or Table 3; or the sense strand comprises a nucleotide sequence that differs by 0, 1, or 2 nucleotides from any one of the sense strands set forth in Table 2 or Table 3; or the RNAi agent comprises a duplex set forth in any one of Table 2 or Table 3.

11. The RNAi agent of claim 10, wherein, The RNAi agent comprises a duplex selected from any one of PAT-0001, PAT-0002, PAT-0004, PAT-0006, PAT-0009, PAT-0009.1, PAT-0009.2, PAT-0010, PAT-0010.1, PAT-0010.2, PAT-0014, PAT-0015, PAT-0016, PAT-0023, PAT-0027, PAT-0028, PAT-0029, PAT-0035, PAT-0036, PAT-0037, PAT-0038, PAT-0038.1, PAT-0038.2, PAT-0038.3, PAT-0038.4, PAT-0038.5, PAT-0039, PAT-0040, PAT-0041, PAT-0042, PAT-0045, PAT-0046, PAT-0048, PAT-0052.

12. The RNAi agent according to any one of claims 1-11, wherein, The RNAi agent further comprises at least one ligand attached to one or more of the 5' end of the antisense strand, the 3' end of the antisense strand, the 5' end of the sense strand, and the 3' end of the sense strand; the ligand is a GalNAc derivative; more preferably the ligand is one or more GalNAc derivatives attached via a single-stranded, double-stranded, or triple-stranded branched linker.

13. The RNAi agent of claim 12, wherein, The RNAi agent comprises a compound having the structure shown below: wherein Nu represents a nucleic acid consisting of the sense strand and the antisense strand.

14. An isolated cell comprising the RNAi agent of any one of claims 1-13.

15. A pharmaceutical composition comprising the RNAi agent of any one of claims 1-13.

16. A method of inhibiting expression of C3 complement in a cell, the method comprising: contacting the cell with the RNAi agent of any one of claims 1-13 or the pharmaceutical composition of claim 15 to inhibit expression of C3 complement in the cell.

17. Use of the RNAi agent of any one of claims 1-13 or the pharmaceutical composition of claim 15 for any one of: 1) treating and / or preventing a disease associated with C3 complement; 2) manufacturing a medicament for treating and / or preventing a disease associated with C3 complement.

18. The use according to claim 17, wherein, The disease is: (i) a disease associated with enhanced or elevated C3 complement; or (ii) a disease that would benefit from a reduction in expression of C3 complement.

19. The use of claim 17, wherein, The disease is selected from one or more of paroxysmal nocturnal hemoglobinuria, atypical hemolytic uremic syndrome, anti-neutrophil cytoplasmic antibody-associated vasculitis, hereditary angioedema, multiple sclerosis, systemic lupus erythematosus, rheumatoid arthritis, Sjogren's syndrome, membranoproliferative glomerulonephritis, IgA nephropathy, primary membranous nephropathy, C3 glomerulopathy, acute tubular necrosis, focal segmental glomerulosclerosis, diabetic nephropathy, neuromyelitis optica, age-related macular degeneration, Guillain-Barre syndrome, multifocal motor neuropathy, and myasthenia gravis.

20. The use according to any one of claims 17 to 19, wherein, The RNAi agent or the pharmaceutical composition is administered to the subject by subcutaneous administration, intravenous administration, and / or intramuscular administration. The RNAi agent or the pharmaceutical composition is administered to the subject by subcutaneous administration, intravenous administration, and / or intramuscular administration. The RNAi agent or the pharmaceutical composition is administered to the subject by subcutaneous administration, intravenous administration, and / or intr

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