Nucleic acids, drug compositions and complexes, as well as methods for their preparation and use.

Abstract

To provide an siRNA that inhibits xanthine oxidase (XO) gene expression, so that diseases caused by abnormal uric acid metabolism, especially hyperuricemia and gout, can be prevented and treated at the cellular level.SOLUTION: The siRNA contains a sense strand and an antisense strand. Each nucleotide in the siRNA is independently a modified or unmodified nucleotide. Lengths of the sense strand and the antisense strand are the same or different, the length of the sense strand is 19-23 nucleotides, and the length of the antisense strand is 19-26 nucleotides. The sense strand contains a specific nucleotide sequence I. The antisense strand contains a specific nucleotide sequence II. The nucleotide sequence I and the nucleotide sequence II form a substantially reverse complementary or completely reverse complementary double-stranded region. The siRNA inhibits expression of an XO gene in hepatocytes.SELECTED DRAWING: Figure 1C

Description

[Technical Field]

[0001] This disclosure relates to nucleic acids that can suppress xanthine oxidase (XO) gene expression, including nucleic acids This disclosure relates to drug compositions and siRNA complexes. This disclosure further relates to these nucleic acids and drug compositions. This relates to the preparation methods and use of substances and siRNA complexes. [Background technology]

[0002] Gout is a disease directly related to hyperuricemia caused by purine metabolism abnormalities and / or decreased uric acid excretion. It is a disease. Gout has long been a common disease in developed countries such as Europe and the United States, and in World War II Following the World Wars, as countries developed economically, the prevalence of the disease has tended to increase year by year worldwide, and The incidence of gout is trending towards younger people. Currently, there are 12 million gout patients in China.

[0003] Xanthine oxidase (XO) is one of the key target points in treating gout. Yes. By suppressing XO expression, the production of hypoxanthine and guanine is effectively suppressed. Furthermore, the aim is to reduce uric acid production, alleviate the progression of gout, and improve the condition. This can be achieved. By suppressing the expression of the XO gene, diseases caused by uric acid metabolism abnormalities can be treated at the cellular level. In particular, it can prevent and treat hyperuricemia and gout. Small interfering RNA (small RNA interference (RNA interfering, siRNA) is a type of RNA interference. Based on a mechanism called reference (RNAi), any target gene of interest The expression of [the substance] can be suppressed or blocked in a sequence-specific manner, thereby achieving the objective of treating the disease.

[0004] To suppress XO gene expression and develop siRNA drugs to treat diseases caused by uric acid metabolism disorders In this context, appropriate siRNA and its modifications, as well as an effective delivery system, are key technologies. . [Overview of the project] [Problems that the invention aims to solve]

[0005] The inventors of this disclosure provide the following siRNAs and their modified sequences as provided by this disclosure. Having this, the expression of the XO gene can be specifically suppressed, and the drug composition or si The RNA complex can specifically target the liver and suppress the expression of the XO gene in the liver. And, surprisingly, it was discovered that it is possible to treat or prevent diseases caused by uric acid metabolism disorders. And thus, the present invention was completed. [Means for solving the problem]

[0006] In some embodiments, the present disclosure provides siRNA capable of repressing XO gene expression. The siRNA comprises a sense strand and an antisense strand, and each nucleotide of the siRNA is Each nucleotide is independently modified or unmodified, and the sense strand is a nucleo The nucleotide sequence I is included, and the antisense strand includes nucleotide sequence II, and the nucleotide sequence Column I and nucleotide sequence II form a double-stranded region in an inversely complementary manner, at least in part. The nucleotide sequence I and the nucleotide sequence II are as shown in i) to xii) below. It is one of the elements selected from the given array.

[0007] i) The nucleotide sequence I and the nucleotide sequence shown in Sequence ID No. 1 are of equal length. Furthermore, the nucleotide difference is three or less, as shown in the nucleotide sequence II and SEQ ID NO: 2 The nucleotide sequences being selected are those with equal lengths and three or fewer nucleotide differences. 5'-GAGAUGAAGUUCAAGAAUZ1-3'(Sequence ID 1), 5'-Z2AUUCUUGAACUUCAUCUC-3'(Sequence ID 2) However, Z1 is A, Z2 is U, and in the nucleotide sequence I, the position is Z1 The corresponding nucleotide Z3 is included, and the nucleotide sequence II has a position corresponding to Z2. The nucleotide Z4 is the first nucleotide at the 5' end of the antisense strand. It is creotide.

[0008] ii) The nucleotide sequence I and the nucleotide sequence shown in Sequence ID No. 61 are, in terms of length They are equal, and the nucleotide difference is 3 or less, and the nucleotide sequence II and SEQ ID NO: 6 The nucleotide sequences shown in 2 are of equal length and have three or fewer nucleotide differences. , 5'-CAUAACUGGAAUUUGUAAZ5-3' (Sequence ID 61), 5'-Z6UUACAAAUUCCAGUUAUG-3'(Sequence ID 62) However, Z5 is U and Z6 is A, and in the aforementioned nucleotide sequence I, the position is Z5 The corresponding nucleotide Z7 is included, and the nucleotide sequence II has a position corresponding to Z6. The nucleotide Z8 is the first nucleotide at the 5' end of the antisense strand. It is creotide.

[0009] iii) The nucleotide sequence I and the nucleotide sequence shown in Sequence ID No. 121 are, The lengths are equal, the nucleotide difference is 3 or less, and the nucleotide sequence is the same as sequence number II. The nucleotide sequences shown in No. 122 are those of equal length and with three or fewer nucleotide differences. And, 5'-CAUUAUCACAAUUGAGGAZ9-3'(Sequence ID 121), 5'-Z10 UCCUCAAUUGUGAUAAUG-3’(SEQ ID NO: 122) However, Z9 is U, and Z 10 is A. In the nucleotide sequence I, the nucleotide Z corresponding to the position Z9 is included. In the nucleotide sequence II, the nucleotide Z corresponding to the position Z 11 is included. The Z 10 is the first nucleotide at the 5'-end of the antisense strand. 12 is 12 the first nucleotide at the 5'-end of the antisense strand. is the first nucleotide at the 5'-end of the antisense strand.

[0010] iv) The nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 181 have the same length and three or fewer nucleotide differences. The nucleotide sequence II and the nucleotide sequence shown in SEQ ID NO: 182 have the same length and three or fewer nucleotide differences. 5'-GGAUCUCUCUCAGAGUAUZ -3’(SEQ ID NO: 181), 5'-Z 13 -3’(SEQ ID NO: 181), 5'-Z 14 AUACUCUGAGAGAGAUCC-3’(SEQ ID NO: 182) However, Z 13 is U, and Z 14 is A. In the nucleotide sequence I, the nucleotide Z corresponding to the position Z 13 is included. In the nucleotide sequence II, the nucleotide Z corresponding to the position Z1 15 is included. The Z corresponding to the position Z14 16 is included. The Z 16 is the first nucleotide at the 5'-end of the antisense strand.

[0011] v) The nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 241 have the same length and three or fewer nucleotide differences. The nucleotide sequence II and SEQ ID NO: 2 have the same length and three or fewer nucleotide differences. The nucleotide sequences shown in 42 are of equal length and have three or fewer nucleotide differences. the law of nature, 5'-ACAUGGACAACUGCUAUAZ 17 -3'(Sequence ID 241), 5'-Z 18 UAUAGCAGUUGUCCAUGU-3' (Sequence ID 242) However, Z 17 A is Z 18 It is U, and in the nucleotide sequence I, position Z 17 Nucleotide Z corresponding to 19 It includes, and the nucleotide sequence II has position Z1 Nucleotide Z corresponding to 8 20 The above Z 20 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0012] vi) The nucleotide sequence I and the nucleotide sequence shown in Sequence ID No. 301 are, The difference is equal, the nucleotide difference is 3 or less, and the nucleotide sequence II and the sequence number are equal. The nucleotide sequences shown in 302 are of equal length and have three or fewer nucleotide differences. can be, 5'-UAGCAAGCUCUCAGUAUCZ 21 -3'(Sequence ID 301), 5'-Z 22 GAUACUGAGAGCUUGCUA-3' (Sequence ID 302) However, Z 21 A is Z 22 It is U, and in the nucleotide sequence I, position Z 21 Nucleotide Z corresponding to 23 It includes, and the nucleotide sequence II has position Z2 Nucleotide Z corresponding to 2 24 The above Z 24 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0013] vii) The nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 361 are, The lengths are equal, the nucleotide difference is 3 or less, and the nucleotide sequence is the same as sequence number II. The nucleotide sequences shown in No. 362 are those of equal length and with three or fewer nucleotide differences. And, 5'-AUAAGGUUACUUGUGUUGZ 25 -3'(Sequence ID 361), 5'-Z 26 CAACACAAGUAACCUUAU-3' (Sequence ID 362) However, Z 25 A is Z 26 It is U, and in the nucleotide sequence I, position Z 25 Nucleotide Z corresponding to 27 It includes, and the nucleotide sequence II has position Z2 Nucleotide Z corresponding to 6 28 The above Z 28 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0014] viii) The nucleotide sequence I and the nucleotide sequence shown in Sequence ID No. 421 are , the lengths are equal, the nucleotide difference is 3 or less, and the nucleotide sequence II and the sequence The nucleotide sequence shown in number 422 is one of equal length and has no more than three nucleotide differences. It is below, 5'-GAAAAUCACCUAUGAAGAZ 29 -3'(Sequence ID 421), 5'-Z 30 UCUUCAUAGGUGAUUUUC-3'(Sequence ID 422) However, Z 29 A is Z 30 It is U, and in the nucleotide sequence I, position Z 29 Nucleotide Z corresponding to 31 It includes, and the nucleotide sequence II has position Z3 Nucleotide Z corresponds to 0 32 The above Z 32 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0015] ix) The nucleotide sequence I and the nucleotide sequence shown in Sequence ID No. 481 are, The difference is equal, the nucleotide difference is 3 or less, and the nucleotide sequence II and the sequence number are equal. The nucleotide sequences shown in 482 are of equal length and have three or fewer nucleotide differences. can be, 5'-GAUGCUAUAAAGAACAACZ 33 -3'(Sequence ID 481), 5'-Z 34 GUUGUUCUUUAUAGCAUC-3' (Sequence ID 482) However, Z 33 is U, Z 34 is A, and in the nucleotide sequence I, position Z 33 Nucleotide Z corresponding to 35 It includes, and the nucleotide sequence II has position Z3 nucleotide Z corresponding to 4 36 The above Z 36 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0016] x) The nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 541 are, by length They are equal, and the nucleotide difference is 3 or less, and the nucleotide sequence II and SEQ ID NO: 5 The nucleotide sequences shown in 42 are of equal length and have three or fewer nucleotide differences. the law of nature, 5'-GAACAACUCCUUUUAUGGZ 37 -3'(Sequence ID 541), 5'-Z 38 CCAUAAAAGGAGUUGUUC-3' (Sequence ID 542) However, Z 37 A is Z 38 It is U, and in the nucleotide sequence I, position Z 37 Nucleotide Z corresponding to 39 It includes, and the nucleotide sequence II has position Z3 Nucleotide Z corresponding to 8 40 The above Z 40 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0017] xi) The nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 601 are, The difference is equal, the nucleotide difference is 3 or less, and the nucleotide sequence II and the sequence number are equal. The nucleotide sequences shown in 602 are of equal length and have three or fewer nucleotide differences. can be, 5'-CUUGCUCUGAAGUAGAAAZ 41 -3'(Sequence ID 601), 5'-Z 42 AUUUCUACUUCAGAGCAAG-3' (Sequence ID 602) However, Z 41 is U, Z 42 is A, and in the nucleotide sequence I, position Z 41 Nucleotide Z corresponding to 43 It includes, and the nucleotide sequence II has a position Z4 Nucleotide Z corresponding to 2 44 The above Z 44 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0018] xii) The nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 661 are, The lengths are equal, the nucleotide difference is 3 or less, and the nucleotide sequence is the same as sequence number II. The nucleotide sequences shown in code 662 are those of equal length and with three or fewer nucleotide differences. And, 5'-CUUCUUUGCCAUCAAAGAZ 45 -3'(Sequence ID 661), 5'-Z 46 UCUUUGAUGGCAAAGAAG-3' (Sequence ID 662) However, Z 45 is U, Z 46 is A, and in the nucleotide sequence I, position Z 45 Nucleotide Z corresponding to 47 It includes, and the nucleotide sequence II has a position Z4 Nucleotide Z corresponding to 6 48 The above Z 48 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0019] In some embodiments, the Disclosure relates to the siRNA and pharmaceutically acceptable The present invention provides a drug composition containing a carrier.

[0020] In some embodiments, this disclosure relates to the siRNA and said The present invention provides an siRNA complex containing a complex group that binds to the siRNA.

[0021] In some embodiments, the Disclosure relates to the siRNA and / or drug compositions of the Disclosure. and / or siRNA complex, uric acid metabolism disorder or disease or physiological condition due to uric acid metabolism disorder It provides use in the preparation of drugs for the treatment and / or prevention of [condition].

[0022] In some embodiments, the Disclosure relates to the siRNA and / or drug compositions of the Disclosure. and / or administering an effective amount of the siRNA complex to a subject requiring it. , propose methods for treating and / or preventing uric acid metabolism disorders or diseases or physiological conditions caused by uric acid metabolism disorders. To provide.

[0023] In some embodiments, the Disclosure relates to the siRNA and / or drug compositions of the Disclosure. and / or bringing an effective amount of the siRNA complex into contact with hepatocytes, This invention provides a method for suppressing the expression of the XO gene.

[0024] In some embodiments, the Disclosure relates to the siRNA and / or drug compositions of the Disclosure. The kit provides the and / or siRNA complex. Incorporation into this site by citation

[0025] All publications, patents, and patent applications referred to herein are subject to the respective publications, patents or patents. To the same extent that a patent application is specifically and individually incorporated herein by reference, This specification is incorporated herein. [Effects of the Invention]

[0026] The siRNAs, drug compositions, and siRNA complexes provided herein are highly stable. Therefore, it exhibits high XO mRNA inhibitory activity, reduced off-target effects, and / or uric acid subsidence. Diseases or physiological conditions due to abnormal metabolism or uric acid metabolism, especially hyperuricemia and / or gout symptoms. It can be significantly treated or alleviated.

[0027] In some embodiments, the siRNA, drug composition or s provided herein The iRNA complex exhibits excellent target gene repression activity in in vitro cell experiments. Several implementations In this state, the siRNA, pharmaceutical composition or siRNA complex provided by the present disclosure shows at least 20%, 30%, 40%, 50%, 60%, 70%, 80% , 90% or 95% target gene expression inhibition rate in hepatocytes. In some embodiments, the present disclosure provides siRNA that shows high inhibitory activity against XO mRNA in an in vitro psiCHECK system and shows a certain inhibitory effect on any of the XO target sequences at different siRNA concentrations. Particularly, at a concentration of 0.1 nM, the inhibition rate against the target sequence is at least 61.39%, and thus reaches as high as 85.43%. In some embodiments, the siRNA provided by the present disclosure shows high inhibitory activity in CAL-27 cells, and the IC against XO mRNA is 0.037 - 0.3277 μM. In some 50 embodiments, the siRNA complex provided by the present disclosure shows high inhibitory activity in primary mouse hepatocytes, and at an siRNA concentration of 20 nM, the inhibition rate against XO mRNA is at least 78.95%, and thus reaches as high as 88.07%.

[0028] In some embodiments, the siRNA, pharmaceutical composition or s iRNA complex provided by the present disclosure can have higher stability and / or higher activity in vivo. In some embodiments, the siRNA, pharmaceutical composition or siRNA complex provided by the present disclosure shows at least 20%, 30%, 40%, 50%, 60% , 70%, 80%, 90% or 95% target gene expression inhibition rate in vivo. In some embodiments , the siRNA, pharmaceutical composition or siRNA complex provided by the present disclosure show at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 95% XO gene expression inhibition rate in vivo. In some embodiments, the siRNA, drug composition or siRNA complex provided by the present disclosure shows at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 95% hepatic XO gene expression inhibition rate in vivo. In some embodiments, the siRNA, drug composition or siRNA complex provided by the present disclosure shows at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 95% hepatic XO gene expression inhibition rate in an animal model in vivo. In some embodiments, the siRNA, drug composition or siRNA complex provided by the present disclosure shows at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 95% hepatic XO gene expression inhibition rate in a human subject in vivo. In some embodiments, at an siRNA concentration of 3 mg / kg, the siRNA complex provided by the present disclosure shows an inhibition rate of 70.9 - 76.2% against XO mRNA expression in mice

[0029] In some embodiments, the siRNA, drug composition or siRNA complex provided by the present disclosure does not show a significant off-target effect. The off- target effect may be, for example, inhibition of the normal expression of a non- target gene. When the binding / suppression of off-target gene expression is 50%, 40%, 30%, 20% or 10% lower than the on-target gene effect, the off-target

[0030] Thus, the siRNA, drug composition and siRNA complex provided by this disclosure are , suppressing the expression of the XO gene, and uric acid metabolism abnormalities or diseases or physiological conditions caused by uric acid metabolism abnormalities It can effectively treat and / or prevent disease, and has promising prospects for application.

[0031] Other features and advantages of this disclosure will be discussed in the embodiment for carrying out the invention, as described later. I will explain in detail. [Brief explanation of the drawing]

[0032] [Figure 1A-1F] This is a dose-effect curve fitted according to the relative expression levels of XO mRNA in in vitro CAL-27 cells after transfection with different siRNAs. [Figure 2] This is a histogram of the relative expression levels of XO mRNA in primary mouse hepatocytes after transfection with different siRNAs. [Figure 3] This is a scatter plot of the relative expression levels of XO mRNA in mice after administration of different siRNA complexes at a dose of 3 mg / kg. [Modes for carrying out the invention]

[0033] The embodiments for carrying out the invention of this disclosure will be described in detail below. The modes for carrying out the invention are merely for illustrative or interpretive purposes of this disclosure. It should be understood that this is not intended to be restrictive.

[0034] In this specification, XO mRNA refers to Genbank registration number NM_000379 This refers to mRNA having the sequence shown in .3. Furthermore, unless otherwise specified, this disclosure refers to The term "target gene" used in this context refers to the gene that transcribes the above-mentioned XO mRNA. The term "target mRNA" refers to the above-mentioned XO mRNA.

[0035] <Definition> In the context, unless otherwise specified, the capital letters C, G, U, and A represent the nucleotide base sequences, the lowercase letter m represents that one nucleotide adjacent to the left of the letter m is a methoxy-modified nucleotide, the lowercase letter f represents that one nucleotide adjacent to the left of the letter f is a fluoro-modified nucleotide, the lowercase letter s represents that the two nucleotides adjacent to the left and right of the letter s are linked by a phosphorothioate group, P1 represents that one nucleotide adjacent to the right of the P1 is a 5'-phosphate nucleotide or a 5'-phosphate analog-modified nucleotide, the combined letter VP represents that one nucleotide adjacent to the right of the combined letter VP is a vinyl phosphate ester-modified nucleotide, the combined letter Ps represents that one nucleotide adjacent to the right of the combined letter Ps is a phosphorothioate ester-modified nucleotide, and the capital letter P represents that one nucleotide adjacent to the right of the letter P is a 5'-phosphate nucleotide.

[0036] In the context, the above-mentioned "fluoro-modified nucleotide" refers to a nucleotide in which the 2'-hydroxy group of the ribose group of the nucleotide is replaced by fluorine, and the "non-fluoro-modified nucleotide" refers to a nucleotide in which the 2'-hydroxy group of the ribose group of the nucleotide is replaced by a non-fluorine group or a nucleotide analog. The "nucleotide analog" refers to a substance that can replace a nucleotide in a nucleic acid, such as an adenine ribonucleotide. Guanine ribonucleotide, cytosine ribonucleotide, uracil ribonucleotide or This refers to a group with a different structure from thymine deoxyribonucleotide, such as isonucleotides. Bridged nucleotides (BNA) or acyclic nucleotides There is a creotide. The aforementioned "methoxy-modified nucleotide" refers to the 2'-hydro ribose group. This refers to a nucleotide in which the xy group is replaced with a methoxy group.

[0037] In the context of this specification, the terms “complementary” and “reverse complementary” are used interchangeably. This is also well known to those skilled in the art, that is, in a double-stranded nucleic acid molecule, the bases of one strand This means that the base on one strand pairs complementaryly with the base on the other strand. In DNA, pre The RNA base adenine (A) is always replaced by the pyrimidine base thymine (T) (or, RNA In this case, uracil (U) pairs with guanine (C), which is a purine base, and always pi It pairs with the limidine base cytosine (G). Each base pair consists of one purine and one It contains two pyrimidines. Adenine on one chain is always present on the other chain (thymine or uracil). ) and when guanine always pairs with cytosine, the two chains are complementary. It is also thought that the sequence of the chain in question can be estimated from the sequence of its complementary chain. In this field, "mismatch" refers to a situation in double-stranded nucleic acids where the bases at corresponding positions are complementary. It means that it does not exist in opposition to the target.

[0038] In context, unless otherwise specified, "basically inversely complementary" means two related nuclei. This refers to the presence of three or fewer base mismatches between rheotide sequences, and is "effectively inversely complementary." "This refers to the presence of one or fewer base mismatches between two nucleotide sequences. "Perfectly reverse complementary" means that there is no base mismatch between the two nucleotide sequences. It refers to.

[0039] In this context, a "nucleotide difference" refers to a difference between one nucleotide sequence and the other nucleotide sequence. The existence of "difference" means that the type of base at the same position of the nucleotide has changed in the former compared to the latter. This refers to the fact that, for example, when one nucleotide base in the latter is A, the same as in the former If the corresponding nucleotide base at the same position is U, C, G, or T, then at that position 2 Nucleotide differences have been recognized between two nucleotide sequences. In the application form, a baseless nucleotide or its equivalent is used instead of the nucleotide at the original position. When using this method, it is also possible that a nucleotide difference occurred at that position.

[0040] In this context, in particular the siRNA, drug composition, or method for preparing the siRNA complex of the present disclosure. When explaining, unless otherwise specified, the nucleoside monomer (nucleosid e-monomer) refers to the nucleo in the siRNA or siRNA complex being prepared. Depending on the type and order of the nucleotides, modified or unmodified nucleotides are used in the solid-phase synthesis of phosphoramidites. Cleoside phosphoramidite monomer (unmodified or modified) d RNA phosphoramidites. RNA phosphoramid (tes is sometimes called Nucleoside phosphoramidites) This refers to phosphoramidite solid-phase synthesis, a method known to those skilled in the art for RNA synthesis. Yes. All nucleoside monomers used in this disclosure are commercially available. ru.

[0041] In the context of this specification, unless otherwise specified, “compound” means each having a specific function. This refers to the covalent bonding between two or more chemical parts, and accordingly, A "complex" is a compound formed by the covalent bonding between its various chemical parts. This refers to a "siRNA complex" which is a complex of one or more chemical parts that have a specific function. This refers to a compound formed by covalent binding to siRNA. In the following text, the present disclosure An siRNA complex is sometimes simply called a "complex." The term siRNA complex can be used in context. The general term for siRNA complexes, and the siRNA complexes represented by formulas (305) and (307). A general term for the organism, or an siRNA complex represented by formulas (305), (307), and (308). It is understood to be a body. In the context of this specification, "composite molecule" is understood to be a body formed by a reaction. It can be compounded with siRNA and ultimately form the siRNA complex of the present disclosure. It should be understood that it is a specific compound.

[0042] As used herein, “any” or “any” means the events or events described below. The situation may or may not occur, and the description may or may not be an event or situation. This refers to both cases where it occurs and cases where it does not. For example, "arbitrarily substituted" alkyl This includes "alkyl" and "substituted alkyl" as defined in the following text: 1 or more With respect to any group containing substituents, these groups are sterically impractical and synthetically unfeasible. It is not intended to introduce any substitutions or substitution patterns that are and / or inherently unstable. If not, it will be understood by those skilled in the art.

[0043] As used herein, "alkyl" means a linear chain having a specific number of carbon atoms and a fractional group. This refers to a branched chain, and the aforementioned specific number is usually 1 to 20 carbon atoms, for example, 1 to 10 carbon atoms. A child is a carbon atom consisting of 1 to 8 or 1 to 6 carbon atoms. For example, C1-C6 alkyl groups have 1 to 6 carbon atoms. This includes linear and branched alkyl groups of carbon atoms. It refers to alkyl residues having a specific number of carbon atoms. In this case, it is intended to include all branched and linear forms having that number of carbon atoms. Therefore, for example, "butyl" includes n-butyl, sec-butyl, isobutyl and tert -This means it contains butyl, and "propyl" includes n-propyl and isopropyl. Alkylenes are a subset of alkyls, similar to alkyls but with two bonding sites. It refers to the residue that does the following.

[0044] As used herein, "alkenyl" means at least one carbon-carbon double bond This refers to an unsaturated branched or linear alkyl chain having a carbon-carbon double bond, wherein the parent alkyl group It is obtained by removing one hydrogen molecule from an adjacent carbon atom of the group. The double bond may be in a cis or trans configuration. Typical alkenyls include vinyl and ,prop-1-en-1-il,prop-1-en-2-il,prop-2-en-1- Propenyls such as yl(allyl), propa-2-en-2-yl, and buta-1-en-1- Il, buta-1-en-2-yl, 2-methylpropa-1-en-1-yl, buta-2- En-1-yl, buta-2-en-2-yl, buta-1,3-dien-1-yl, buta- This includes, but is not limited to, but but also includes, but is not limited to, but but also includes, a certain implementation form. In this state, the alkenyl has 2 to 20 carbon atoms, but in other embodiments, It has 2 to 10, 2 to 8, or 2 to 6 carbon atoms. Alkenylenes are alkenyl This is a subset, referring to residues that are the same as alkenils but have two binding sites.

[0045] As used herein, "alkynyl" means at least one carbon-carbon triple bond This refers to an unsaturated branched or linear alkyl group having a carbon-carbon triple bond, where the carbon-carbon triple bond is a hermaphrodite. It is obtained by removing two hydrogen molecules from adjacent carbon atoms of the kill molecule. Typical A Lukinyl is composed of ethinyl, propane-1-in-1-yl, and propane-2-in-1- Propynnyl, including yl, and buta-1-in-1-yl, buta-1-in-3-yl, buta- This includes, but is not limited to, but but also includes, 3-in-1-yl and other butynyl compounds. In one embodiment... In other embodiments, alkynyl has 2 to 20 carbon atoms, but in other embodiments, 2 to 1 They have 0, 2-8, or 2-6 carbon atoms. Alkynylenes are a subset of alkynyls. It is the same as alkynyl, but refers to a residue that has two binding sites.

[0046] As used herein, "alkoxy" means a specific number of carbon atoms linked by oxygen bridges. This refers to alkyl groups of elementary atoms, such as methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, pentyloxy, 2-pentyl Oxy, isopentyloxy, neopentyloxy, hexyloxy, 2-hexyloxy Examples include 3-hexyloxy and 3-methylpentyloxy. Alkoxy compounds are usually 1- It has 10, 1-8, 1-6, or 1-4 carbon atoms bonded by oxygen bridges. .

[0047] As used herein, "aryl" refers to an aromatic monocyclic or polycyclic hydrocarbon ring system. This refers to a group formed by removing a hydrogen atom from a ring carbon atom, derived from the aforementioned aromatic compound. A cyclic or polycyclic hydrocarbon ring system contains only hydrogen and 6 to 18 carbon atoms. At least one of the rings in the ring system is perfectly unsaturated, that is, it follows Hückel's theory. It includes cyclic and delocalized (4n+2)π-electron systems. Examples of aryls include phenyl and fluorine. Arylenes include, but are not limited to, renyl and naphthyl groups. This is a subset, referring to residues that are the same as aryl residues but have two binding sites.

[0048] As used herein, "halogen substituent" or "halo" means fluoro, chloro The term "halogen" refers to bromine or iodine, and includes fluorine, chlorine, bromine, or iodine. .

[0049] As used herein, "alkyl halogenate" means a specific number of carbon atoms. This is an alkyl group defined above, which is substituted with multiple halogen atoms up to the maximum allowable number. It refers to the group. Examples of alkyl halides include trifluoromethyl and difluoromethyl. This includes, but is not limited to, 2-fluoroethyl or pentafluoroethyl.

[0050] A "heterocyclic group" is a group consisting of 2 to 12 carbon atoms and 1 atom selected from nitrogen, oxygen, or sulfur. This refers to a stable 3- to 18-membered non-aromatic cyclic group containing six heteroatoms. Unless otherwise specified, heterocyclic groups are monocyclic, dicyclic, tricyclic, or tetracyclic systems, and are fused or bridged rings. A ring system may be included. Heteroatoms in the heterocyclic group may be optionally oxidized. 1 or Multiple nitrogen atoms (if present) are optionally quaternized. Heterocyclic groups are partially saturated or complete. It is totally saturated. Heterocyclic groups can bond to the rest of the molecule via any atom in the ring. Yes, it is possible. Examples of such heterocyclic groups include dioxanyl and thiophenyl[1,3]di Sulfonyl (thienyl[1,3]dithianyl), decahydroisoquinolinyl Lu, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, Morpholinyl, octahydroindole, octahydroisoindole, 2-oxapipe Rajinil, 2-oxapiperidil, 2-oxapyrrolidinil, oxazolidinil, piperi Zill, piperazinil, 4-piperidonil, pyrrolidinil, pyrazolidil, quinuclidinil , thiazolidinyl, tetrahydrofuryl, trithianyl, te Trahydropyranil, thiomorpholinyl, thiamol folinyl (thiamorpholinyl), 1-oxothiomorpholinyl (1-ox o-thiomorpholinyl) and 1,1-dioxothiomorpholinyl (1,1 This includes, but is not limited to, -dioxo-thiomorpholinyl.

[0051] A "heteroaryl" is a compound consisting of 2 to 17 carbon atoms and selected nitrogen, oxygen, and sulfur. This refers to a group derived from a 3-18 membered aromatic ring radical containing 1-6 heteroatoms. As used herein, heteroaryl is a monocyclic, dicyclic, tricyclic, or tetracyclic system. Also, at least one of the rings in this ring system is completely unsaturated, that is, it is Hucke It contains a cyclic delocalized (4n+2)π-electron system that follows the theory of cyclic delocalization. Heteroaryls are fused rings or It contains a bridging ring system. The heteroatoms in the heteroaryl are optionally oxidized. One or more The nitrogen atom (if present) is optionally quaternized. The heteroaryl is any in the ring It is bonded to the rest of the molecule via atoms. An example of a heteroaryl is azepinine. Lu, acridinyl, benzimidazolyl, benzoindole, 1,3-benzodioxazol Lyl, benzofuryl, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazol Lil, benzo[b][1,4]dioxepinyl pinyl), benzo[b][1,4]oxazinyl (benzo[b][1,4]ox azinyl), 1,4-benzodioxanyl, Benzonaphthofuranil, benzoxazolyl, benzodioxolyl olyl), benzodioxinyl, benzopyranyl, Nzopyronil, benzofuryl, benzofuranol, benzothiophenyl, benzothieno[ 3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[1,2- a] Pyridyl, carbazolyl, cinnolinyl, cyclopenta[d ]pyrimidinyl, 6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d ] Pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl (5,6-dihydro benzo[h]quinazolinyl), 5,6-dihydrobenzo[h]cinnoly Nyl(5,6-dihydrobenzo[h]cinnolinyl), 6,7-dihydrobenzo[h]cinnolinyl Dro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofri Lu, dibenzothiophenyl, furyl, furanol, flo[3,2-c]pyridyl, 5,6 ,7,8,9,10-Hexahydrocycloocta[d]pyrimidinyl, 5,6,7,8, 9,10-Hexahydrocycloocta[d]pyridazinyl, 5,6,7,8,9,10- Hexahydrocycloocta[d]pyridyl, isothiazolyl, imidazolyl, indazoli Indazolyl, indole, isoindole, indolinyl, isoindri Nyl, Isoquinolyl, Indolizinyl, Isoxazolyl, 5,8-methano-5,6,7,8-tetrahydroquinazolinyl(5,8-methano (-5,6,7,8-tetrahydroquinazolinyl), naphthilidinyl (naphthyridinyl), 1,6-naphthyridinyl (1,6-naphth yridinonyl), oxadiazolyl, 2-oxoazepinyl pinyl), oxazolyl, oxiranyl, 5,6,6a,7, 8,9,10,10a-Octahydrobenzo[H]quinazolinyl,1-phenyl-1H- Pyrrolyl, phenazinil, phenothiazinil, phenoxadinil, phthalazinil (pht halazinyl, pteridinyl, prinyl, pyrrolyl, Pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridyl, pyrido[3,2-d]pyrimidinyl Limidinil, Pyrid[3,4-d]pyrimidinil, Pyrazinil, Pyrimidinil, Pyridadi Nyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinoli Lu, tetrahydroquinolyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7, 8-Tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6,7,8,9 -Tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl, 5 ,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiacia Zolyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidin Lu, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pridinyl (thien o[2,3-c]pridinyl) and thiophenyl (thiophenyl / thi This includes, but is not limited to, enyl.

[0052] Various hydroxy protecting groups can be used in this disclosure. Generally, protecting groups are The chemical functionality can be made insensitive to specific reaction conditions, and the rest of the molecule can be substantially It can be added to and removed from the functionality in the molecule without damaging it. A typical hydroxy protecting group is Beaucage et al., Tetrahedron 19 92,48,2223~2311, and Greene and Wuts, Protec tive Groups in Organic Synthesis,Chapter 2,2d ed., John Wiley & Sons, New York, 1991 Disclosed in and incorporated herein by reference as a whole. In some embodiments, the protecting group is stable under basic conditions but under acidic conditions. It can be removed. In some embodiments, hydroxyl can be used herein. Non-exclusive examples of protecting groups include dimethoxytrityl (DMT) and monomethoxytrityl. , 9-phenylxanthene-9-yl (Pixyl) or 9-(p-methoxyphenyl) Contains xanthene-9-yl (Mox). In some embodiments, used herein Non-exclusive examples of hydroxy protecting groups that can be formed include Tr (trityl) and MMTr (4-methyl). Toxytrityl), DMTr(4,4'-dimethoxytrityl) or TMTr(4,4' Contains ,4''-trimethoxytrityl).

[0053] As used herein, the term "subject" refers to any animal, for example, a mammal. or marsupials. Subjects of this disclosure include humans and non-human primates (e.g., rhesus). (Little macaques or other species of macaques), mice, pigs, horses, donkeys, cattle, sheep, rats or any type of poultry, including but not limited to these.

[0054] As used herein, “treatment” means a method for obtaining a beneficial or desired outcome. This includes, but is not limited to, therapeutic effects. "Therapeutic effect" means eradicating the underlying disorder being treated. This means improvement. Furthermore, the therapeutic effect indicates that the subject is still suffering from the underlying disability. Despite the possibility of experiencing it, one or more physiological symptoms associated with the potential disorder are rooted in By eliminating or improving the condition, improvement is observed in the subject.

[0055] As used herein, “prevention” means a method for obtaining a beneficial or desired outcome. This includes, but is not limited to, preventive effects. In order to obtain a "preventive effect," the diagnosis of the disease in question Although a diagnosis may not have been made, siRNA, siRNA complexes, or drug compositions are particularly Administration to subjects at risk of developing certain diseases, or in cases where one or more pathological symptoms of a disease are present. It can be administered to the reported subjects.

[0056] In certain contexts, this disclosure provides first to twelfth siRNAs capable of suppressing XO gene expression. We will provide them. The following will explain them in detail, one by one.

[0057] The siRNAs of this disclosure include a nucleotide group as a basic structural unit, and the nucleo It is known to those skilled in the art that the tide group includes a phosphate group, a ribose group, and a base, so here I will omit the explanation.

[0058] <First siRNA> According to this disclosure, the siRNA may be a first siRNA.

[0059] The first siRNA comprises a sense strand and an antisense strand, and each of the first siRNA Each nucleotide is independently modified or unmodified, and the sense strand is The nucleotide sequence I is included, the antisense strand includes the nucleotide sequence II, and the nucleotide sequence Cleotide sequence I and nucleotide sequence II are double-stranded in a reverse complementary manner, at least in part. The region is formed, and the nucleotide sequence I and the nucleotide sequence shown in Sequence ID No. 1 are The lengths are equal, the nucleotide difference is 3 or less, and the nucleotide sequence is the same as sequence number II. The nucleotide sequences shown in No. 2 are those with equal lengths and three or fewer nucleotide differences. ru. 5'-GAGAUGAAGUUCAAGAAUZ1-3'(Sequence ID 1), 5'-Z2AUUCUUGAACUUCAUCUC-3'(Sequence ID 2) However, Z1 is A, Z2 is U, and in the nucleotide sequence I, the position is Z1 The corresponding nucleotide Z3 is included, and the nucleotide sequence II has a position corresponding to Z2. The nucleotide Z4 is the first nucleotide at the 5' end of the antisense strand. It is creotide.

[0060] In this context, "corresponding positions" means that from the same end of a nucleotide sequence, the nucleotides correspond to each other. This refers to the same position in a sequence. For example, nucleotide 1 at the 3' end of nucleotide sequence I. The nucleotide in the eye corresponds to the 1st nucleotide at the 3' end of sequence number 1. It is leotide.

[0061] In some embodiments, the sense strand comprises only nucleotide sequence I, and The antisense strand contains only nucleotide sequence II.

[0062] In some embodiments, the nucleotide sequence I and the nucleotide shown in SEQ ID NO: 1 The nucleotide difference between the ocidal sequence and / or the nucleotide sequence is one or less, and / or the nucleotide sequence If there is one or fewer nucleotide differences between the nucleotide sequences shown in column II and sequence number 2 be.

[0063] In some embodiments, the nucleotide sequence II and the nucleotide shown in SEQ ID NO: 2 The nucleotide differences between the rheotide sequence and the rheotide sequence include differences at the position of Z4, where Z4 is A, C or It is selected from G. In some embodiments, the nucleotide difference is at position Z4. The difference lies in the position, where Z4 is selected from A, C, or G. In some embodiments, Z3 is a nucleotide complementary to Z4. siRNA with the above nucleotide differences. These siRNAs possess high target mRNA repression capabilities, and siRNAs containing these nucleotide differences also have high target mRNA repression capabilities. This information is within the scope of protection of this disclosure.

[0064] In some embodiments, the nucleotide sequence I and the nucleotide sequence II are , basically inversely complementary, substantially inversely complementary, or completely inversely complementary, the aforementioned basically inversely complementary A mismatch refers to the presence of three or fewer base mismatches between two nucleotide sequences. The term "substantially reverse complementary" means that there is one or fewer base mismatches between the two nucleotide sequences. This refers to the presence of a base mismatch between two nucleotide sequences, and being completely inversely complementary means that there is no base mismatch between the two nucleotide sequences. It refers to something that is not present.

[0065] In some embodiments, nucleotide sequence I is the nucleo shown in SEQ ID NO: 3 The nucleotide sequence is the nucleotide sequence shown in Sequence ID No. 4. ru. 5'-GAGAUGAAGUUCAAGAAUZ3-3'(Sequence ID 3), 5'-Z4AUUCUUGAACUUCAUCUC-3'(Sequence ID 4) However, the aforementioned Z4 is the first nucleotide at the 5' end of the antisense strand, and Z4 Z3 is selected from A, U, G, or C, and Z3 is a nucleotide complementary to Z4, and In one embodiment, Z3 is A and Z4 is U.

[0066] The sense chain and the antisense chain may have the same or different lengths, and the length of the sense chain may be It consists of 19-23 nucleotides, and the antisense strand is 19-26 nucleotides long. Thus, the sense strand to antisense strand length ratio of the siRNA provided in this disclosure. 19 / 19, 19 / 20, 19 / 21, 19 / 22, 19 / 23, 19 / 24, 19 / 25, 19 / 26, 20 / 20, 20 / 21, 20 / 22, 20 / 23, 20 / 24, 2 0 / 25, 20 / 26, 21 / 20, 21 / 21, 21 / 22, 21 / 23, 21 / 24 , 21 / 25, 21 / 26, 22 / 20, 22 / 21, 22 / 22, 22 / 23, 22 / 24, 22 / 25, 22 / 26, 23 / 20, 23 / 21, 23 / 22, 23 / 23, 2 It may be 3 / 24, 23 / 25, or 23 / 26. In some embodiments, The length ratio of the sense strand to the antisense strand of the siRNA is 19 / 21, 21 / 23, or 23 / It is 25.

[0067] In some embodiments, the sense strand further comprises nucleotide sequence III. The antisense strand further comprises nucleotide sequence IV and nucleotide sequence III The nucleotide sequence IV has a length of 1 to 4 nucleotides, and the nucleotide sequence IV has a length of 1 to 4 nucleotides. Sequence III and nucleotide sequence IV are of equal length and substantially inversely complementary or complete. It is inversely complementary to the above, and the nucleotide sequence III is at the 5' end of the nucleotide sequence I. The nucleotide sequence IV is bound to the 3' end of the nucleotide sequence II. In some embodiments, the nucleotide sequence IV and the second nucleotide sequence The sequence is substantially inversely complementary or completely inversely complementary, and is related to the second nucleotide sequence. It is adjacent to the 5' end of the nucleotide sequence shown in SEQ ID NO: 1 in the target mRNA, This refers to a nucleotide sequence whose length is equal to that of nucleotide sequence IV.

[0068] In some embodiments, nucleotide sequence III and nucleotide sequence IV are The length of each is 1 nucleotide, and the base of nucleotide sequence III is U, The base of creotide sequence IV is A, and in this case, the length ratio of the sense strand to the antisense strand is It is 20 / 20, or nucleotide sequences III and IV are both 2 nucleotides in length. It is an Otid, and the base sequence of nucleotide sequence III is from the 5' end to the 3' end. It is UU, and the base sequence of nucleotide sequence IV is AA, and in this case, the sense strand and AN The length ratio of the cysnes chains is 21 / 21, or nucleotide sequences III and IV are long Each of them consists of 3 nucleotides, and the nucleotide arrangement is such that it moves from the 5' end to the 3' end. The base sequence of column III is AUU, and the base sequence of nucleotide sequence IV is AAU. At this time, the length ratio of the sense chain to the antisense chain is 22 / 22, or nucleoty Sequences III and IV are both 4 nucleotides long, and from the 5' end to the 3' end... Towards the next step, the base sequence of nucleotide sequence III is CAUU, and nucleotide sequence IV The base sequence is AAUG, and in this case, the length ratio of the sense strand to the antisense strand is 23 / 2. 3. In some embodiments, the nucleotide sequence III and the nucleotide sequence Column IV is 2 nucleotides long, and from the 5' end to the 3' end, it is nucleotide The base sequence of sequence III is UU, and the base sequence of nucleotide sequence IV is AA. In this case, the length ratio of the sense chain to the antisense chain is 21 / 21.

[0069] In some embodiments, nucleotide sequences III and nucleotide IV are complete Since they are inversely complementary, given the bases of nucleotide sequence III, nucleotide The bases of sequence IV are also determined.

[0070] <Second siRNA> According to this disclosure, the siRNA may be a second siRNA.

[0071] The second siRNA comprises a sense strand and an antisense strand, and each of the second siRNA Each nucleotide is independently modified or unmodified, and the sense strand is The nucleotide sequence I is included, the antisense strand includes the nucleotide sequence II, and the nucleotide sequence Cleotide sequence I and nucleotide sequence II are double-stranded in a reverse complementary manner, at least in part. The region is formed, and the nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 61 are different , the lengths are equal, the nucleotide difference is 3 or less, and the nucleotide sequence II and the sequence The nucleotide sequence shown in number 62 is one that is equal in length and has three or fewer nucleotide differences. That is the case. 5'-CAUAACUGGAAUUUGUAAZ5-3' (Sequence ID 61), 5'-Z6UUACAAAUUCCAGUUAUG-3'(Sequence ID 62) However, Z5 is U and Z6 is A, and in the aforementioned nucleotide sequence I, the position is Z5 The corresponding nucleotide Z7 is included, and the nucleotide sequence II has a position corresponding to Z6. The nucleotide Z8 is the first nucleotide at the 5' end of the antisense strand. It is creotide.

[0072] In some embodiments, the sense strand comprises only nucleotide sequence I, and The antisense strand contains only nucleotide sequence II.

[0073] In some embodiments, the nucleotide sequence I and the nucleotide shown in SEQ ID NO: 61 The nucleotide difference between the rheotide sequence and the rheotide sequence is one or less, and / or the nucleotide There is no difference of one nucleotide between the nucleotide sequence shown in sequence II and sequence number 62. It is below.

[0074] In some embodiments, the nucleotide sequence II and the nucleotide sequence shown in SEQ ID NO: 62 are used. Nucleotide differences between the sequence and the creotide sequence include differences at the Z8 position, where Z8 is U, C Or selected from G. In some embodiments, the nucleotide difference is Z8 This is a positional difference, where Z8 is selected from U, C, or G. In some embodiments, Z7 is a nucleotide complementary to Z8. SIRN with the above nucleotide differences. A has high target mRNA suppression ability, and siRNAs containing these nucleotide differences also , within the scope of protection of this disclosure.

[0075] In some embodiments, the nucleotide sequence I and the nucleotide sequence II are They are basically inversely complementary, substantially inversely complementary, or completely inversely complementary.

[0076] In some embodiments, nucleotide sequence I is the nucleotide shown in SEQ ID NO: 63 The ocidal sequence is the nucleotide sequence II shown in Sequence ID No. 64. That is the case. 5'-CAUAACUGGAAUUUGUAAZ7-3' (SEQ ID NO: 63), 5'-Z8UUACAAAUUCCAGUUAUG-3'(Sequence ID 64) However, the aforementioned Z8 is the first nucleotide at the 5' end of the antisense strand, and Z8 Z7 is selected from A, U, G, or C, and Z7 is a nucleotide complementary to Z8, and In one embodiment, Z7 is U and Z8 is A.

[0077] The sense chain and the antisense chain may have the same or different lengths, and the length of the sense chain may be It consists of 19-23 nucleotides, and the antisense strand is 19-26 nucleotides long. .

[0078] In some embodiments, the sense strand further comprises nucleotide sequence III. The antisense strand further comprises nucleotide sequence IV and nucleotide sequence III The nucleotide sequence IV has a length of 1 to 4 nucleotides, and the nucleotide sequence IV has a length of 1 to 4 nucleotides. Sequence III and nucleotide sequence IV are of equal length and substantially inversely complementary or complete. It is inversely complementary to the above, and the nucleotide sequence III is at the 5' end of the nucleotide sequence I. The nucleotide sequence IV is bound to the 3' end of the nucleotide sequence II. The nucleotide sequence IV and the second nucleotide sequence are substantially inversely complementary or complete. It is completely inversely complementary, and the second nucleotide sequence is the sequence number in the target mRNA. Adjacent to the 5' end of the nucleotide sequence shown in No. 61, the length is the same as the nucleotide sequence I This refers to a nucleotide sequence that is equal to the selected one.

[0079] In some embodiments, nucleotide sequence III and nucleotide sequence IV are The length of each is 1 nucleotide, and the base of nucleotide sequence III is A, The base of creotide sequence IV is U, and in this case, the length ratio of the sense strand to the antisense strand is It is 20 / 20, or nucleotide sequences III and IV are both 2 nucleotides in length. It is an Otid, and the base sequence of nucleotide sequence III is from the 5' end to the 3' end. It is AA, and the base sequence of nucleotide sequence IV is UU, and in this case the sense strand and AN The length ratio of the cysnes chains is 21 / 21, or nucleotide sequences III and IV are long Each of them consists of 3 nucleotides, and the nucleotide arrangement is such that it moves from the 5' end to the 3' end. The base sequence of column III is UAA, and the base sequence of nucleotide sequence IV is UUA. At this time, the length ratio of the sense chain to the antisense chain is 22 / 22, or nucleoty Sequences III and IV are both 4 nucleotides long, and from the 5' end to the 3' end... Towards the next step, the base sequence of nucleotide sequence III is GUAA, and nucleotide sequence IV The base sequence is UUAC, and in this case, the length ratio of the sense strand to the antisense strand is 23 / 2. 3. In some embodiments, the nucleotide sequence III and the nucleotide sequence Column IV is 2 nucleotides long, and from the 5' end to the 3' end, it is nucleotide The base sequence of sequence III is AA, and the base sequence of nucleotide sequence IV is UU. In this case, the length ratio of the sense chain to the antisense chain is 21 / 21.

[0080] In some embodiments, nucleotide sequences III and nucleotide IV are complete Since they are inversely complementary, given the bases of nucleotide sequence III, nucleotide The bases of sequence IV are also determined.

[0081] <Third siRNA> According to this disclosure, the siRNA may be a third siRNA.

[0082] The third siRNA comprises a sense strand and an antisense strand, and each of the third siRNA Each nucleotide is independently modified or unmodified, and the sense strand is The nucleotide sequence I is included, the antisense strand includes the nucleotide sequence II, and the nucleotide sequence Cleotide sequence I and nucleotide sequence II are double-stranded in a reverse complementary manner, at least in part. A region is formed, and the nucleotide sequence I and the nucleotide sequence shown in Sequence ID No. 121 It has the same length, a nucleotide difference of 3 or less, and is paired with the nucleotide sequence II. The nucleotide sequence shown in column number 122 is of equal length and has three nucleotide differences. The following applies: 5'-CAUUAUCACAAUUGAGGAZ9-3'(Sequence ID 121), 5'-Z 10 UCCUCAAUUGUGAUAAUG-3' (Sequence ID 122) However, Z9 is U, Z 10 It is A, and in the nucleotide sequence I, the position is Z9 Nucleotide Z corresponding to 11 It includes, and the nucleotide sequence II has position Z 10 to Corresponding nucleotide Z 12 The above Z 12 The 5' end of the antisense chain It is the first nucleotide.

[0083] In some embodiments, the sense strand comprises only nucleotide sequence I, and The antisense strand contains only nucleotide sequence II.

[0084] In some embodiments, the nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 121 The nucleotide difference between the cleotide sequence and / or the nucleotide sequence is one or less, and / or the nucleotide sequence There is a nucleotide difference of 1 between sequence II and the nucleotide sequence shown in sequence number 122. It is one or less.

[0085] In some embodiments, the nucleotide sequence II and sequence number 122 are shown The nucleotide difference between the nucleotide sequence and the nucleotide sequence is Z 12 Including the difference in position, Z 12 but Selected from U, C, or G. In some embodiments, the nucleotide difference is Z12 is the difference at the position of Z 12 is selected from U, C or G. In some embodiments Z 11 is a nucleotide complementary to Z 12 . The siRNA having the above nucleotide difference has a high target mRNA inhibitory ability, and siRNA containing these nucleotide differences is also within the protection scope of the present disclosure.

[0086] In some embodiments, the nucleotide sequence I and the nucleotide sequence II are basically reverse complementary, substantially reverse complementary or completely reverse complementary.

[0087] In some embodiments, the nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 123, and the nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO: 124 . 5'-CAUUAUCACAAUUGAGGAZ -3'(SEQ ID NO: 123), 11 5'-Z UCCUCAAUUGUGAUAAUG-3'(SEQ ID NO: 124) 12 However, the Z is the first nucleotide at the 5' end of the antisense strand, and Z 12 is selected from A, U, G or C, and Z 12 is a nucleotide complementary to Z 11 , and in some embodiments, Z 12 is U, and Z is A. 11 12 12

[0088] The sense strand and the antisense strand may have the same or different lengths, the length of the sense strand is 19-23 nucleotides, and the length of the antisense strand is 19-26 nucleotides .

[0089] ​ In some embodiments, the sense strand further comprises nucleotide sequence III. The antisense strand further comprises nucleotide sequence IV and nucleotide sequence III The nucleotide sequence IV has a length of 1 to 4 nucleotides, and the nucleotide sequence IV has a length of 1 to 4 nucleotides. Sequence III and nucleotide sequence IV are of equal length and substantially inversely complementary or complete. It is inversely complementary to the above, and the nucleotide sequence III is at the 5' end of the nucleotide sequence I. The nucleotide sequence IV is bound to the 3' end of the nucleotide sequence II. The nucleotide sequence IV and the second nucleotide sequence are substantially inversely complementary or complete. It is completely inversely complementary, and the second nucleotide sequence is the sequence number in the target mRNA. Adjacent to the 5' end of the nucleotide sequence shown in No. 121, and the length is the same as the nucleotide sequence. This refers to a nucleotide sequence equal to IV.

[0090] In some embodiments, the nucleotide sequence III follows the 5'-3' direction. Nucleotide sequence IV and nucleotide sequence I are both 1 nucleotide in length, and nucleotide sequence I The base of nucleotide II is C, and the base of nucleotide sequence IV is G. In this case, the sense strand and The antisense strand length ratio is 20 / 20, or nucleotide sequences III and IV are Each has a length of 2 nucleotides, and from the 5' end to the 3' end, the nucleoti The base sequence of sequence III is GC, and the base sequence of nucleotide sequence IV is GC. At this time, the length ratio of the sense chain to the antisense chain is 21 / 21, or nucleoty Sequences III and IV are both 3 nucleotides long, and from the 5' end to the 3' end... Towards the right, the base sequence of nucleotide sequence III is AGC, and the base sequence of nucleotide sequence IV is The base sequence is GCU, and in this case, the length ratio of the sense strand to the antisense strand is 22 / 22. Alternatively, nucleotide sequences III and IV are both 4 nucleotides long. From the 5' end to the 3' end, the base sequence of nucleotide sequence III is CAGC. Furthermore, the base sequence of nucleotide sequence IV is GCUG, and in this case, the sense strand and antisequence The nucleotide chain length ratio is 23 / 23. In some embodiments, the nucleotide arrangement Column III and nucleotide sequence IV are 2 nucleotides long, from the 5' end to the 3' end. Towards the end, the base sequence of nucleotide sequence III is GC, and nucleotide sequence IV The base sequence is GC, and in this case the length ratio of the sense strand to the antisense strand is 21 / 21. be.

[0091] In some embodiments, nucleotide sequences III and nucleotide IV are complete Since they are inversely complementary, given the bases of nucleotide sequence III, nucleotide The bases of sequence IV are also determined.

[0092] <The fourth siRNA> According to this disclosure, the siRNA may be a fourth siRNA.

[0093] The fourth siRNA comprises a sense strand and an antisense strand, and each of the fourth siRNA Each nucleotide is independently modified or unmodified, and the sense strand is The nucleotide sequence I is included, the antisense strand includes the nucleotide sequence II, and the nucleotide sequence Cleotide sequence I and nucleotide sequence II are double-stranded in a reverse complementary manner, at least in part. A region is formed, and the nucleotide sequence I and the nucleotide sequence shown in Sequence ID No. 181 It has the same length, a nucleotide difference of 3 or less, and is paired with the nucleotide sequence II. The nucleotide sequence shown in column number 182 is of equal length and has three nucleotide differences. The following applies: 5'-GGAUCUCUCUCAGAGUAUZ 13 -3'(Sequence ID 181), 5'-Z 14 AUACUCUGAGAGAGAUCC-3' (Sequence ID 182) However, Z 13 is U, Z 14 is A, and in the nucleotide sequence I, position Z 13 Nucleotide Z corresponding to 15 It includes, and the nucleotide sequence II has position Z1 nucleotide Z corresponding to 4 16 The above Z 16 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0094] In some embodiments, the sense strand comprises only nucleotide sequence I, and The antisense strand contains only nucleotide sequence II.

[0095] In some embodiments, the nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 181 The nucleotide difference between the cleotide sequence and / or the nucleotide sequence is one or less, and / or the nucleotide sequence There is a nucleotide difference of 1 between sequence II and the nucleotide sequence shown in sequence number 182. It is one or less.

[0096] In some embodiments, the nucleotide sequence II and sequence number 182 are shown The nucleotide difference between the nucleotide sequence and the nucleotide sequence is Z 16 Including the difference in position, Z16 but Selected from U, C, or G. In some embodiments, the nucleotide difference is Z 16 This is the difference at the position of Z 16 is selected from U, C, or G. Several implementation forms In state, Z 15 is, Z 16 It is a complementary nucleotide. The above nucleotide differences The siRNA possesses high target mRNA repression ability and contains these nucleotide differences. siRNA is also within the scope of protection of this disclosure.

[0097] In some embodiments, the nucleotide sequence I and the nucleotide sequence II are They are basically inversely complementary, substantially inversely complementary, or completely inversely complementary.

[0098] In some embodiments, nucleotide sequence I is the nucleotide shown in SEQ ID NO: 183 The rheotide sequence is the nucleotide sequence II shown in SEQ ID NO: 184. It is an array. 5'-GGAUCUCUCUCAGAGUAUZ 15 -3'(Sequence ID 183), 5'-Z 16 AUACUCUGAGAGAGAUCC-3' (Sequence ID 184) However, the aforementioned Z 16 Z is the first nucleotide at the 5' end of the antisense strand. 16 is selected from A, U, G, or C, and Z 15 is, Z 16 It is a complementary nucleotide. In some embodiments, Z 15 is U, Z 16 It is A.

[0099] The sense chain and the antisense chain may have the same or different lengths, and the length of the sense chain may be It consists of 19-23 nucleotides, and the antisense strand is 19-26 nucleotides long. .

[0100] In some embodiments, the sense strand further comprises nucleotide sequence III. The antisense strand further comprises nucleotide sequence IV and nucleotide sequence III The nucleotide sequence IV has a length of 1 to 4 nucleotides, and the nucleotide sequence IV has a length of 1 to 4 nucleotides. Sequence III and nucleotide sequence IV are of equal length and substantially inversely complementary or complete. It is inversely complementary to the above, and the nucleotide sequence III is at the 5' end of the nucleotide sequence I. The nucleotide sequence IV is bound to the 3' end of the nucleotide sequence II. The nucleotide sequence IV and the second nucleotide sequence are substantially inversely complementary or complete. It is completely inversely complementary, and the second nucleotide sequence is the sequence number in the target mRNA. Adjacent to the 5' end of the nucleotide sequence shown in No. 181, and the length is the same as the nucleotide sequence. This refers to a nucleotide sequence equal to IV.

[0101] In some embodiments, the nucleotide sequence III follows the 5'-3' direction. Nucleotide sequence IV and nucleotide sequence I are both 1 nucleotide in length, and nucleotide sequence I The base of nucleotide sequence II is A, and the base of nucleotide sequence IV is U. In this case, the sense strand and The antisense strand length ratio is 20 / 20, or nucleotide sequences III and IV are Each has a length of 2 nucleotides, and from the 5' end to the 3' end, the nucleoti The base sequence of sequence III is CA, and the base sequence of nucleotide sequence IV is UG. At this time, the length ratio of the sense chain to the antisense chain is 21 / 21, or nucleoty Sequences III and IV are both 3 nucleotides long, and from the 5' end to the 3' end... Towards the right, the base sequence of nucleotide sequence III is CCA, and the base sequence of nucleotide sequence IV is The base sequence is UGG, and in this case, the length ratio of the sense strand to the antisense strand is 22 / 22. Alternatively, nucleotide sequences III and IV are both 4 nucleotides long. From the 5' end to the 3' end, the base sequence of nucleotide sequence III is CCCA. Furthermore, the base sequence of nucleotide sequence IV is UGGG, and in this case, the sense strand and antisequence The nucleotide chain length ratio is 23 / 23. In some embodiments, the nucleotide arrangement Column III and nucleotide sequence IV are 2 nucleotides long, from the 5' end to the 3' end. Towards the end, the base sequence of nucleotide sequence III is CA, and nucleotide sequence IV The base sequence is UG, and in this case the length ratio of the sense strand to the antisense strand is 21 / 21. be.

[0102] In some embodiments, nucleotide sequences III and nucleotide IV are complete Since they are inversely complementary, given the bases of nucleotide sequence III, nucleotide The bases of sequence IV are also determined.

[0103] <The fifth siRNA> According to this disclosure, the siRNA may be a fifth siRNA.

[0104] The fifth siRNA comprises a sense strand and an antisense strand, and each of the fifth siRNA Each nucleotide is independently modified or unmodified, and the sense strand is The nucleotide sequence I is included, the antisense strand includes the nucleotide sequence II, and the nucleotide sequence Cleotide sequence I and nucleotide sequence II are double-stranded in a reverse complementary manner, at least in part. A region is formed, and the nucleotide sequence I and the nucleotide sequence shown in Sequence ID No. 241 It has the same length, a nucleotide difference of 3 or less, and is paired with the nucleotide sequence II. The nucleotide sequence shown in column number 242 is of equal length and has three nucleotide differences. The following applies: 5'-ACAUGGACAACUGCUAUAZ 17 -3'(Sequence ID 241), 5'-Z 18 UAUAGCAGUUGUCCAUGU-3' (Sequence ID 242) However, Z 17 A is Z 18 It is U, and in the nucleotide sequence I, position Z 17 Nucleotide Z corresponding to 19 It includes, and the nucleotide sequence II has position Z1 Nucleotide Z corresponding to 8 20 The above Z 20 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0105] In some embodiments, the sense strand comprises only nucleotide sequence I, and The antisense strand contains only nucleotide sequence II.

[0106] In some embodiments, the nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 241 are used. The nucleotide difference between the cleotide sequence and / or the nucleotide sequence is one or less, and / or the nucleotide sequence There is a nucleotide difference of 1 between sequence II and the nucleotide sequence shown in sequence number 242. It is one or less.

[0107] In some embodiments, the nucleotide sequence II and sequence number 242 are shown The nucleotide difference between the nucleotide sequence and the nucleotide sequence is Z 20 Including the difference in position, Z 20 but Selected from A, C, or G. In some embodiments, the nucleotide difference is Z 20 This is the difference at the position of Z 20 The selected form is A, C, or G. In state, Z 19 is, Z 20 It is a complementary nucleotide. The above nucleotide differences The siRNA possesses high target mRNA repression ability and contains these nucleotide differences. siRNA is also within the scope of protection of this disclosure.

[0108] In some embodiments, the nucleotide sequence I and the nucleotide sequence II are They are basically inversely complementary, substantially inversely complementary, or completely inversely complementary.

[0109] In some embodiments, nucleotide sequence I is the nucleotide shown in SEQ ID NO: 243 The rheotide sequence is the nucleotide sequence II shown in Sequence ID No. 244. It is an array. 5'-ACAUGGACAACUGCUAUAZ 19 -3'(Sequence ID 243), 5'-Z 20 UAUAGCAGUUGUCCAUGU-3' (Sequence ID 244) However, the aforementioned Z 20 Z is the first nucleotide at the 5' end of the antisense strand. 20 is selected from A, U, G, or C, and Z 19 is, Z 20 It is a complementary nucleotide. In some embodiments, Z 19 A is Z 20 It is U.

[0110] The sense chain and the antisense chain may have the same or different lengths, and the length of the sense chain may be It consists of 19-23 nucleotides, and the antisense strand is 19-26 nucleotides long. .

[0111] In some embodiments, the sense strand further comprises nucleotide sequence III. The antisense strand further comprises nucleotide sequence IV and nucleotide sequence III The nucleotide sequence IV has a length of 1 to 4 nucleotides, and the nucleotide sequence IV has a length of 1 to 4 nucleotides. Sequence III and nucleotide sequence IV are of equal length and substantially inversely complementary or complete. It is inversely complementary to the above, and the nucleotide sequence III is at the 5' end of the nucleotide sequence I. The nucleotide sequence IV is bound to the 3' end of the nucleotide sequence II. The nucleotide sequence IV and the second nucleotide sequence are substantially inversely complementary or complete. It is completely inversely complementary, and the second nucleotide sequence is the sequence number in the target mRNA. Adjacent to the 5' end of the nucleotide sequence shown in No. 241, and the length is the same as the nucleotide sequence. This refers to a nucleotide sequence equal to IV.

[0112] In some embodiments, the nucleotide sequence III follows the 5'-3' direction. Nucleotide sequence IV and nucleotide sequence I are both 1 nucleotide in length, and nucleotide sequence I The base of nucleotide II is C, and the base of nucleotide sequence IV is G. In this case, the sense strand and The antisense strand length ratio is 20 / 20, or nucleotide sequences III and IV are Each has a length of 2 nucleotides, and from the 5' end to the 3' end, the nucleoti The base sequence of nucleotide sequence III is CC, and the base sequence of nucleotide sequence IV is GG. At this time, the length ratio of the sense chain to the antisense chain is 21 / 21, or nucleoty Sequences III and IV are both 3 nucleotides long, and from the 5' end to the 3' end... Towards the end, the base sequence of nucleotide sequence III is UCC, and the base sequence of nucleotide sequence IV is The base sequence is GGA, and in this case, the length ratio of the sense strand to the antisense strand is 22 / 22. Alternatively, nucleotide sequences III and IV are both 4 nucleotides long. From the 5' end to the 3' end, the base sequence of nucleotide sequence III is UUCC. Furthermore, the base sequence of nucleotide sequence IV is GGAA, and in this case, the sense strand and antisequence The nucleotide chain length ratio is 23 / 23. In some embodiments, the nucleotide arrangement Column III and nucleotide sequence IV are 2 nucleotides long, from the 5' end to the 3' end. Towards the end, the base sequence of nucleotide sequence III is CC, and nucleotide sequence IV The base sequence is GG, and in this case, the length ratio of the sense strand to the antisense strand is 21 / 21. be.

[0113] In some embodiments, nucleotide sequences III and nucleotide IV are complete Since they are inversely complementary, given the bases of nucleotide sequence III, nucleotide The bases of sequence IV are also determined.

[0114] <The 6th siRNA> According to this disclosure, the siRNA may be a sixth siRNA.

[0115] The sixth siRNA comprises a sense strand and an antisense strand, and each of the sixth siRNA Each nucleotide is independently modified or unmodified, and the sense strand is The nucleotide sequence I is included, the antisense strand includes the nucleotide sequence II, and the nucleotide sequence Cleotide sequence I and nucleotide sequence II are double-stranded in a reverse complementary manner, at least in part. A region is formed, and the nucleotide sequence I and the nucleotide sequence shown in Sequence ID No. 301 It has the same length, a nucleotide difference of 3 or less, and is paired with the nucleotide sequence II. The nucleotide sequence shown in column number 302 is of equal length and has three nucleotide differences. The following applies: 5'-UAGCAAGCUCUCAGUAUCZ 21 -3'(Sequence ID 301), 5'-Z 22 GAUACUGAGAGCUUGCUA-3' (Sequence ID 302) However, Z 21 A is Z 22 It is U, and in the nucleotide sequence I, position Z 21 Nucleotide Z corresponding to 23 It includes, and the nucleotide sequence II has position Z2 Nucleotide Z corresponding to 2 24 The above Z 24 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0116] In some embodiments, the sense strand comprises only nucleotide sequence I, and The antisense strand contains only nucleotide sequence II.

[0117] In some embodiments, the nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 301 The nucleotide difference between the cleotide sequence and / or the nucleotide sequence is one or less, and / or the nucleotide sequence There is a nucleotide difference of 1 between sequence II and the nucleotide sequence shown in sequence number 302. It is one or less.

[0118] In some embodiments, the nucleotide sequence II and sequence number 302 are shown The nucleotide difference between the nucleotide sequence and the nucleotide sequence is Z 24 Including the difference in position, Z 24 but Selected from A, C, or G. In some embodiments, the nucleotide difference is Z 24 This is the difference at the position of Z 24 The selected form is A, C, or G. In state, Z 23 is, Z 24 It is a complementary nucleotide. The above nucleotide differences The siRNA possesses high target mRNA repression ability and contains these nucleotide differences. siRNA is also within the scope of protection of this disclosure.

[0119] In some embodiments, the nucleotide sequence I and the nucleotide sequence II are They are basically inversely complementary, substantially inversely complementary, or completely inversely complementary.

[0120] In some embodiments, nucleotide sequence I is the nucleotide shown in SEQ ID NO: 303 The rheotide sequence is the nucleotide sequence II shown in SEQ ID NO: 304. It is an array. 5'-UAGCAAGCUCUCAGUAUCZ 23 -3'(Sequence ID 303), 5'-Z 24 GAUACUGAGAGCUUGCUA-3' (Sequence ID 304) However, the aforementioned Z 24 Z is the first nucleotide at the 5' end of the antisense strand. 24 is selected from A, U, G, or C, and Z 23 is, Z 24 It is a complementary nucleotide. In some embodiments, Z 23A is Z 24 It is U.

[0121] The sense chain and the antisense chain may have the same or different lengths, and the length of the sense chain may be It consists of 19-23 nucleotides, and the antisense strand is 19-26 nucleotides long. .

[0122] In some embodiments, the sense strand further comprises nucleotide sequence III. The antisense strand further comprises nucleotide sequence IV and nucleotide sequence III The nucleotide sequence IV has a length of 1 to 4 nucleotides, and the nucleotide sequence IV has a length of 1 to 4 nucleotides. Sequence III and nucleotide sequence IV are of equal length and substantially inversely complementary or complete. It is inversely complementary to the above, and the nucleotide sequence III is at the 5' end of the nucleotide sequence I. The nucleotide sequence IV is bound to the 3' end of the nucleotide sequence II. The nucleotide sequence IV and the second nucleotide sequence are substantially inversely complementary or complete. It is completely inversely complementary, and the second nucleotide sequence is the sequence number in the target mRNA. Adjacent to the 5' end of the nucleotide sequence shown in No. 301, and the length is the same as the nucleotide sequence. This refers to a nucleotide sequence equal to IV.

[0123] In some embodiments, the nucleotide sequence III follows the 5'-3' direction. Nucleotide sequence IV and nucleotide sequence I are both 1 nucleotide in length, and nucleotide sequence I The base of nucleotide II is C, and the base of nucleotide sequence IV is G. In this case, the sense strand and The antisense strand length ratio is 20 / 20, or nucleotide sequences III and IV are Each has a length of 2 nucleotides, and from the 5' end to the 3' end, the nucleoti The base sequence of nucleotide sequence III is CC, and the base sequence of nucleotide sequence IV is GG. At this time, the length ratio of the sense chain to the antisense chain is 21 / 21, or nucleoty Sequences III and IV are both 3 nucleotides long, and from the 5' end to the 3' end... Towards the end, the base sequence of nucleotide sequence III is GCC, and the base sequence of nucleotide sequence IV is The base sequence is GGC, and in this case, the length ratio of the sense strand to the antisense strand is 22 / 22. Alternatively, nucleotide sequences III and IV are both 4 nucleotides long. From the 5' end to the 3' end, the base sequence of nucleotide sequence III is UGCC. Furthermore, the base sequence of nucleotide sequence IV is GGCA, and in this case, the sense strand and antisequence The nucleotide chain length ratio is 23 / 23. In some embodiments, the nucleotide arrangement Column III and nucleotide sequence IV are 2 nucleotides long, from the 5' end to the 3' end. Towards the end, the base sequence of nucleotide sequence III is CC, and nucleotide sequence IV The base sequence is GG, and in this case, the length ratio of the sense strand to the antisense strand is 21 / 21. be.

[0124] In some embodiments, nucleotide sequences III and nucleotide IV are complete Since they are inversely complementary, given the bases of nucleotide sequence III, nucleotide The bases of sequence IV are also determined.

[0125] <The 7th siRNA> According to this disclosure, the siRNA may be a seventh siRNA.

[0126] The seventh siRNA comprises a sense strand and an antisense strand, and each of the seventh siRNA Each nucleotide is independently modified or unmodified, and the sense strand is The nucleotide sequence I is included, the antisense strand includes the nucleotide sequence II, and the nucleotide sequence Cleotide sequence I and nucleotide sequence II are double-stranded in a reverse complementary manner, at least in part. A region is formed, and the nucleotide sequence I and the nucleotide sequence shown in Sequence ID No. 361 It has the same length, a nucleotide difference of 3 or less, and is paired with the nucleotide sequence II. The nucleotide sequence shown in column number 362 is of equal length and has three nucleotide differences. The following applies: 5'-AUAAGGUUACUUGUGUUGZ 25 -3'(Sequence ID 361), 5'-Z 26 CAACACAAGUAACCUUAU-3' (Sequence ID 362) However, Z 25 G is Z 26 is C, and in the nucleotide sequence I, position Z 25 Nucleotide Z corresponding to 27 It includes, and the nucleotide sequence II has position Z2 Nucleotide Z corresponding to 6 28 The above Z 28 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0127] In some embodiments, the sense strand comprises only nucleotide sequence I, and The antisense strand contains only nucleotide sequence II.

[0128] In some embodiments, the nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 361 The nucleotide difference between the cleotide sequence and / or the nucleotide sequence is one or less, and / or the nucleotide sequence There is a nucleotide difference of 1 between sequence II and the nucleotide sequence shown in sequence number 362. It is one or less.

[0129] In some embodiments, the nucleotide sequence II and sequence number 362 are shown The nucleotide difference between the nucleotide sequence and the nucleotide sequence is Z 28 Including the difference in position, Z 28 but Selected from A, U, or G. In some embodiments, the nucleotide difference is Z 28 This is the difference at the position of Z 28 The selected is A, U, or G. Several implementation forms In state, Z 27 is, Z 28 It is a complementary nucleotide. The above nucleotide differences The siRNA possesses high target mRNA repression ability and contains these nucleotide differences. siRNA is also within the scope of protection of this disclosure.

[0130] In some embodiments, the nucleotide sequence I and the nucleotide sequence II are They are basically inversely complementary, substantially inversely complementary, or completely inversely complementary.

[0131] In some embodiments, nucleotide sequence I is the nucleotide shown in SEQ ID NO: 363 The rheotide sequence is the nucleotide sequence II shown in SEQ ID NO: 364. It is an array. 5'-AUAAGGUUACUUGUGUUGZ 27 -3'(Sequence ID 363), 5'-Z 28 CAACACAAGUAACCUUAU-3' (Sequence ID 364) However, the aforementioned Z 28 Z is the first nucleotide at the 5' end of the antisense strand. 28 is selected from A, U, G, or C, and Z 27 is, Z 28It is a complementary nucleotide. In some embodiments, Z 27 G is Z 28 C is C.

[0132] The sense chain and the antisense chain may have the same or different lengths, and the length of the sense chain may be It consists of 19-23 nucleotides, and the antisense strand is 19-26 nucleotides long. .

[0133] In some embodiments, the sense strand further comprises nucleotide sequence III. The antisense strand further comprises nucleotide sequence IV and nucleotide sequence III The nucleotide sequence IV has a length of 1 to 4 nucleotides, and the nucleotide sequence IV has a length of 1 to 4 nucleotides. Sequence III and nucleotide sequence IV are of equal length and substantially inversely complementary or complete. It is inversely complementary to the above, and the nucleotide sequence III is at the 5' end of the nucleotide sequence I. The nucleotide sequence IV is bound to the 3' end of the nucleotide sequence II. In some embodiments, the nucleotide sequence IV and the second nucleotide sequence The sequence is substantially inversely complementary or completely inversely complementary, and is related to the second nucleotide sequence. This is adjacent to the 5' end of the nucleotide sequence shown in SEQ ID NO: 361 in the target mRNA. This refers to a nucleotide sequence whose length is equal to that of nucleotide sequence IV.

[0134] In some embodiments, the nucleotide sequence III follows the 5'-3' direction. Nucleotide sequence IV and nucleotide sequence I are both 1 nucleotide in length, and nucleotide sequence I The base of nucleotide II is G, and the base of nucleotide sequence IV is C. In this case, the sense strand and The antisense strand length ratio is 20 / 20, or nucleotide sequences III and IV are Each has a length of 2 nucleotides, and from the 5' end to the 3' end, the nucleoti The base sequence of nucleotide sequence III is GG, and the base sequence of nucleotide sequence IV is CC. At this time, the length ratio of the sense chain to the antisense chain is 21 / 21, or nucleoty Sequences III and IV are both 3 nucleotides long, and from the 5' end to the 3' end... Towards the right, the base sequence of nucleotide sequence III is AGG, and the base sequence of nucleotide sequence IV is The base sequence is CCU, and in this case, the length ratio of the sense strand to the antisense strand is 22 / 22. Alternatively, nucleotide sequences III and IV are both 4 nucleotides long. From the 5' end to the 3' end, the base sequence of nucleotide sequence III is AAGG. Furthermore, the base sequence of nucleotide sequence IV is CCUU, and in this case, the sense strand and antisequence The nucleotide chain length ratio is 23 / 23. In some embodiments, the nucleotide arrangement Column III and nucleotide sequence IV are 2 nucleotides long, from the 5' end to the 3' end. Towards the end, the base sequence of nucleotide sequence III is GG, and nucleotide sequence IV The base sequence is CC, and in this case, the length ratio of the sense strand to the antisense strand is 21 / 21. be.

[0135] In some embodiments, nucleotide sequences III and nucleotide IV are complete Since they are inversely complementary, given the bases of nucleotide sequence III, nucleotide The bases of sequence IV are also determined.

[0136] <8th siRNA> According to this disclosure, the siRNA may be an eighth siRNA.

[0137] The eighth siRNA comprises a sense strand and an antisense strand, and each of the eighth siRNA Each nucleotide is independently modified or unmodified, and the sense strand is The nucleotide sequence I is included, the antisense strand includes the nucleotide sequence II, and the nucleotide sequence Cleotide sequence I and nucleotide sequence II are double-stranded in a reverse complementary manner, at least in part. A region is formed, and the nucleotide sequence I and the nucleotide sequence shown in Sequence ID No. 421 It has the same length, a nucleotide difference of 3 or less, and is paired with the nucleotide sequence II. The nucleotide sequence shown in column number 422 is of equal length and has three nucleotide differences. The following applies: 5'-GAAAAUCACCUAUGAAGAZ 29 -3'(Sequence ID 421), 5'-Z 30 UCUUCAUAGGUGAUUUUC-3'(Sequence ID 422) However, Z 29 A is Z 30 It is U, and in the nucleotide sequence I, position Z 29 Nucleotide Z corresponding to 31 It includes, and the nucleotide sequence II has position Z3 Nucleotide Z corresponds to 0 32 The above Z 32 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0138] In some embodiments, the sense strand comprises only nucleotide sequence I, and The antisense strand contains only nucleotide sequence II.

[0139] In some embodiments, the nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 421 The nucleotide difference between the cleotide sequence and / or the nucleotide sequence is one or less, and / or the nucleotide sequence There is a nucleotide difference of 1 between sequence II and the nucleotide sequence shown in sequence number 422. It is one or less.

[0140] In some embodiments, the nucleotide sequence II and sequence number 422 are shown The nucleotide difference between the nucleotide sequence and the nucleotide sequence is Z 32 Including the difference in position, Z 32 but Selected from A, C, or G. In some embodiments, the nucleotide difference is Z 32 This is the difference at the position of Z 32 The selected form is A, C, or G. In state, Z 31 is, Z 32 It is a complementary nucleotide. The above nucleotide differences The siRNA possesses high target mRNA repression ability and contains these nucleotide differences. siRNA is also within the scope of protection of this disclosure.

[0141] In some embodiments, the nucleotide sequence I and the nucleotide sequence II are They are basically inversely complementary, substantially inversely complementary, or completely inversely complementary.

[0142] In some embodiments, nucleotide sequence I is the nucleotide shown in SEQ ID NO: 423 It is a rheotide sequence, and the nucleotide sequence II is the nucleo shown in Sequence ID No. 424. It is a CHIDO sequence. 5'-GAAAAUCACCUAUGAAGAZ 31 -3'(Sequence ID 423), 5'-Z 32 UCUUCAUAGGUGAUUUUC-3'(Sequence ID 424) However, the aforementioned Z 32Z is the first nucleotide at the 5' end of the antisense strand. 32 is selected from A, U, G, or C, and Z 31 is, Z 32 It is a complementary nucleotide. In some embodiments, Z 31 A is Z 32 It is U.

[0143] The sense chain and the antisense chain may have the same or different lengths, and the length of the sense chain may be It consists of 19-23 nucleotides, and the antisense strand is 19-26 nucleotides long. .

[0144] In some embodiments, the sense strand further comprises nucleotide sequence III. The antisense strand further comprises nucleotide sequence IV and nucleotide sequence III The nucleotide sequence IV has a length of 1 to 4 nucleotides, and the nucleotide sequence IV has a length of 1 to 4 nucleotides. Sequence III and nucleotide sequence IV are of equal length and substantially inversely complementary or complete. It is inversely complementary to the above, and the nucleotide sequence III is at the 5' end of the nucleotide sequence I. The nucleotide sequence IV is bound to the 3' end of the nucleotide sequence II. In some embodiments, the nucleotide sequence IV and the second nucleotide sequence The sequence is substantially inversely complementary or completely inversely complementary, and is related to the second nucleotide sequence. This is adjacent to the 5' end of the nucleotide sequence shown in SEQ ID NO: 421 in the target mRNA. This refers to a nucleotide sequence whose length is equal to that of nucleotide sequence IV.

[0145] In some embodiments, the nucleotide sequence III follows the 5'-3' direction. Nucleotide sequence IV and nucleotide sequence I are both 1 nucleotide in length, and nucleotide sequence I The base of nucleotide II is U, and the base of nucleotide sequence IV is A. In this case, the sense strand and The antisense strand length ratio is 20 / 20, or nucleotide sequences III and IV are Each has a length of 2 nucleotides, and from the 5' end to the 3' end, the nucleoti The base sequence of nucleotide sequence III is GU, and the base sequence of nucleotide sequence IV is AC. At this time, the length ratio of the sense chain to the antisense chain is 21 / 21, or nucleoty Sequences III and IV are both 3 nucleotides long, and from the 5' end to the 3' end... Towards the end, the base sequence of nucleotide sequence III is GGU, and the base sequence of nucleotide sequence IV is The base sequence is ACC, and in this case, the length ratio of the sense strand to the antisense strand is 22 / 22. Alternatively, nucleotide sequences III and IV are both 4 nucleotides long. From the 5' end towards the 3' end, the base sequence of nucleotide sequence III is GGGU. Furthermore, the base sequence of nucleotide sequence IV is ACCC, and in this case, the sense strand and antisequence The nucleotide chain length ratio is 23 / 23. In some embodiments, the nucleotide arrangement Column III and nucleotide sequence IV are 2 nucleotides long, from the 5' end to the 3' end. Towards the end, the base sequence of nucleotide sequence III is GU, and nucleotide sequence IV The base sequence is AC, and in this case the length ratio of the sense strand to the antisense strand is 21 / 21. be.

[0146] In some embodiments, nucleotide sequences III and nucleotide IV are complete Since they are inversely complementary, given the bases of nucleotide sequence III, nucleotide The bases of sequence IV are also determined.

[0147] <The 9th siRNA> According to this disclosure, the siRNA may be a ninth siRNA.

[0148] The 9th siRNA comprises a sense strand and an antisense strand, and each of the 9th siRNA Each nucleotide is independently modified or unmodified, and the sense strand is The nucleotide sequence I is included, the antisense strand includes the nucleotide sequence II, and the nucleotide sequence Cleotide sequence I and nucleotide sequence II are double-stranded in a reverse complementary manner, at least in part. A region is formed, and the nucleotide sequence I and the nucleotide sequence shown in Sequence ID No. 481 It has the same length, a nucleotide difference of 3 or less, and is paired with the nucleotide sequence II. The nucleotide sequence shown in column number 482 is of equal length and has three nucleotide differences. The following applies: 5'-GAUGCUAUAAAGAACAACZ 33 -3'(Sequence ID 481), 5'-Z 34 GUUGUUCUUUAUAGCAUC-3' (Sequence ID 482) However, Z 33 is U, Z 34 is A, and in the nucleotide sequence I, position Z 33 Nucleotide Z corresponding to 35 It includes, and the nucleotide sequence II has position Z3 nucleotide Z corresponding to 4 36 The above Z 36 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0149] In some embodiments, the sense strand comprises only nucleotide sequence I, and The antisense strand contains only nucleotide sequence II.

[0150] In some embodiments, the nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 481 The nucleotide difference between the cleotide sequence and / or the nucleotide sequence is one or less, and / or the nucleotide sequence There is a nucleotide difference of 1 between sequence II and the nucleotide sequence shown in sequence number 482. It is one or less.

[0151] In some embodiments, the nucleotide sequence II and sequence number 482 are shown The nucleotide difference between the nucleotide sequence and the nucleotide sequence is Z 36 Including the difference in position, Z 36 but Selected from U, C, or G. In some embodiments, the nucleotide difference is Z 36 This is the difference at the position of Z 36 is selected from U, C, or G. Several implementation forms In state, Z 35 is, Z 36 It is a complementary nucleotide. The above nucleotide differences The siRNA possesses high target mRNA repression ability and contains these nucleotide differences. siRNA is also within the scope of protection of this disclosure.

[0152] In some embodiments, the nucleotide sequence I and the nucleotide sequence II are They are basically inversely complementary, substantially inversely complementary, or completely inversely complementary.

[0153] In some embodiments, nucleotide sequence I is the nucleotide shown in SEQ ID NO: 483 The rheotide sequence is the nucleotide sequence II shown in SEQ ID NO: 484. It is an array. 5'-GAUGCUAUAAAGAACAACZ 35-3'(Sequence ID 483), 5'-Z 36 GUUGUUCUUUAUAGCAUC-3' (Sequence ID 484) However, the aforementioned Z 36 Z is the first nucleotide at the 5' end of the antisense strand. 36 is selected from A, U, G, or C, and Z 35 is, Z 36 It is a complementary nucleotide. In some embodiments, Z 35 is U, Z 36 It is A.

[0154] The sense chain and the antisense chain may have the same or different lengths, and the length of the sense chain may be It consists of 19-23 nucleotides, and the antisense strand is 19-26 nucleotides long. .

[0155] In some embodiments, the sense strand further comprises nucleotide sequence III. The antisense strand further comprises nucleotide sequence IV and nucleotide sequence III The nucleotide sequence IV has a length of 1 to 4 nucleotides, and the nucleotide sequence IV has a length of 1 to 4 nucleotides. Sequence III and nucleotide sequence IV are of equal length and substantially inversely complementary or complete. It is inversely complementary to the above, and the nucleotide sequence III is at the 5' end of the nucleotide sequence I. The nucleotide sequence IV is bound to the 3' end of the nucleotide sequence II. In some embodiments, the nucleotide sequence IV and the second nucleotide sequence The sequence is substantially inversely complementary or completely inversely complementary, and is related to the second nucleotide sequence. This is adjacent to the 5' end of the nucleotide sequence shown in SEQ ID NO: 481 in the target mRNA. This refers to a nucleotide sequence whose length is equal to that of nucleotide sequence IV.

[0156] In some embodiments, the nucleotide sequence III follows the 5'-3' direction. Nucleotide sequence IV and nucleotide sequence I are both 1 nucleotide in length, and nucleotide sequence I The base of nucleotide II is G, and the base of nucleotide sequence IV is C. In this case, the sense strand and The antisense strand length ratio is 20 / 20, or nucleotide sequences III and IV are Each has a length of 2 nucleotides, and from the 5' end to the 3' end, the nucleoti The base sequence of sequence III is AG, and the base sequence of nucleotide sequence IV is CU. At this time, the length ratio of the sense chain to the antisense chain is 21 / 21, or nucleoty Sequences III and IV are both 3 nucleotides long, and from the 5' end to the 3' end... Towards the end, the base sequence of nucleotide sequence III is GAG, and the base sequence of nucleotide sequence IV is The base sequence is CUC, and in this case, the length ratio of the sense strand to the antisense strand is 22 / 22. Alternatively, nucleotide sequences III and IV are both 4 nucleotides long. From the 5' end to the 3' end, the base sequence of nucleotide sequence III is UGAG. Furthermore, the base sequence of nucleotide sequence IV is CUCA, and in this case, the sense strand and antisequence The nucleotide chain length ratio is 23 / 23. In some embodiments, the nucleotide arrangement Column III and nucleotide sequence IV are 2 nucleotides long, from the 5' end to the 3' end. Towards the end, the base sequence of nucleotide sequence III is AG, and nucleotide sequence IV The base sequence is CU, and in this case, the length ratio of the sense strand to the antisense strand is 21 / 21. be.

[0157] In some embodiments, nucleotide sequences III and nucleotide IV are complete Since they are inversely complementary, given the bases of nucleotide sequence III, nucleotide The bases of sequence IV are also determined.

[0158] <The 10th siRNA> According to this disclosure, the siRNA may be a tenth siRNA.

[0159] The 10th siRNA comprises a sense strand and an antisense strand. Each nucleotide is independently modified or unmodified, and the sense The strand contains nucleotide sequence I, and the antisense strand contains nucleotide sequence II, The nucleotide sequence I and the nucleotide sequence II are, at least in part, inversely complementary. This chain region is formed, and the nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 541 A row is defined as a sequence of nucleotides of equal length with a nucleotide difference of 3 or less, as described in the nucleotide sequence II. The nucleotide sequence shown in sequence number 542 is of equal length, and the nucleotide difference is There are three or fewer. 5'-GAACAACUCCUUUUAUGGZ 37 -3'(Sequence ID 541), 5'-Z 38 CCAUAAAAGGAGUUGUUC-3' (Sequence ID 542) However, Z 37 A is Z 38 It is U, and in the nucleotide sequence I, position Z 37 Nucleotide Z corresponding to 39 It includes, and the nucleotide sequence II has position Z3 Nucleotide Z corresponding to 8 40 The above Z 40 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0160] In some embodiments, the sense strand comprises only nucleotide sequence I, and The antisense strand contains only nucleotide sequence II.

[0161] In some embodiments, the nucleotide sequence I and the sequence number 541 shown in the sequence number 541 are used. The nucleotide difference between the cleotide sequence and / or the nucleotide sequence is one or less, and / or the nucleotide sequence There is a nucleotide difference of 1 between sequence II and the nucleotide sequence shown in sequence number 542. It is one or less.

[0162] In some embodiments, the nucleotide sequence II and sequence number 542 are shown The nucleotide difference between the nucleotide sequence and the nucleotide sequence is Z 40 Including the difference in position, Z 40 but Selected from A, C, or G. In some embodiments, the nucleotide difference is Z 40 This is the difference at the position of Z 40 The selected form is A, C, or G. In state, Z 39 is, Z 40 It is a complementary nucleotide. The above nucleotide differences The siRNA possesses high target mRNA repression ability and contains these nucleotide differences. siRNA is also within the scope of protection of this disclosure.

[0163] In some embodiments, the nucleotide sequence I and the nucleotide sequence II are They are basically inversely complementary, substantially inversely complementary, or completely inversely complementary.

[0164] In some embodiments, nucleotide sequence I is the nucleotide shown in SEQ ID NO: 543 The rheotide sequence is the nucleotide sequence II shown in SEQ ID NO: 544. It is an array. 5'-GAACAACUCCUUUUAUGGZ 39 -3'(Sequence ID 543), 5'-Z 40 CCAUAAAAGGAGUUGUUC-3' (Sequence ID 544) However, the aforementioned Z 40 Z is the first nucleotide at the 5' end of the antisense strand. 40 is selected from A, U, G, or C, and Z 39 is, Z 40 It is a complementary nucleotide. In some embodiments, Z 39 A is Z 40 It is U.

[0165] The sense chain and the antisense chain may have the same or different lengths, and the length of the sense chain may be It consists of 19-23 nucleotides, and the antisense strand is 19-26 nucleotides long. .

[0166] In some embodiments, the sense strand further comprises nucleotide sequence III. The antisense strand further comprises nucleotide sequence IV and nucleotide sequence III The nucleotide sequence IV has a length of 1 to 4 nucleotides, and the nucleotide sequence IV has a length of 1 to 4 nucleotides. Sequence III and nucleotide sequence IV are of equal length and substantially inversely complementary or complete. It is inversely complementary to the above, and the nucleotide sequence III is at the 5' end of the nucleotide sequence I. The nucleotide sequence IV is bound to the 3' end of the nucleotide sequence II. In some embodiments, the nucleotide sequence IV and the second nucleotide sequence The sequence is substantially inversely complementary or completely inversely complementary, and is related to the second nucleotide sequence. This is adjacent to the 5' end of the nucleotide sequence shown in SEQ ID NO: 541 in the target mRNA. This refers to a nucleotide sequence whose length is equal to that of nucleotide sequence IV.

[0167] In some embodiments, the nucleotide sequence III follows the 5'-3' direction. Nucleotide sequence IV and nucleotide sequence I are both 1 nucleotide in length, and nucleotide sequence I The base of nucleotide sequence II is A, and the base of nucleotide sequence IV is U. In this case, the sense strand and The antisense strand length ratio is 20 / 20, or nucleotide sequences III and IV are Each has a length of 2 nucleotides, and from the 5' end to the 3' end, the nucleoti The base sequence of sequence III is AA, and the base sequence of nucleotide sequence IV is UU. At this time, the length ratio of the sense chain to the antisense chain is 21 / 21, or nucleoty Sequences III and IV are both 3 nucleotides long, and from the 5' end to the 3' end... Towards the end, the base sequence of nucleotide sequence III is AAA, and the base sequence of nucleotide sequence IV is The base sequence is UUU, and in this case, the length ratio of the sense strand to the antisense strand is 22 / 22. Alternatively, nucleotide sequences III and IV are both 4 nucleotides long. From the 5' end to the 3' end, the base sequence of nucleotide sequence III is UAAA. Furthermore, the base sequence of nucleotide sequence IV is UUUA, and in this case, the sense strand and antisequence The nucleotide chain length ratio is 23 / 23. In some embodiments, the nucleotide arrangement Column III and nucleotide sequence IV are 2 nucleotides long, from the 5' end to the 3' end. Towards the end, the base sequence of nucleotide sequence III is AA, and nucleotide sequence IV The base sequence is UU, and in this case, the length ratio of the sense strand to the antisense strand is 21 / 21. be.

[0168] In some embodiments, nucleotide sequences III and nucleotide IV are complete Since they are inversely complementary, given the bases of nucleotide sequence III, nucleotide The bases of sequence IV are also determined.

[0169] <11th siRNA> According to this disclosure, the siRNA may be an eleventh siRNA.

[0170] The 11th siRNA comprises a sense strand and an antisense strand. Each nucleotide is independently modified or unmodified, and the sense The strand contains nucleotide sequence I, and the antisense strand contains nucleotide sequence II, The nucleotide sequence I and the nucleotide sequence II are, at least in part, inversely complementary. This chain region is formed by the nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 601. A row is defined as a sequence of nucleotides of equal length with a nucleotide difference of 3 or less, as described in the nucleotide sequence II. The nucleotide sequence shown in sequence number 602 is of equal length, and the nucleotide difference is There are three or fewer. 5'-CUUGCUCUGAAGUAGAAAZ 41 -3'(Sequence ID 601), 5'-Z 42 AUUUCUACUUCAGAGCAAG-3' (Sequence ID 602) However, Z 41 is U, Z 42 is A, and in the nucleotide sequence I, position Z 41 Nucleotide Z corresponding to 43 It includes, and the nucleotide sequence II has a position Z4 Nucleotide Z corresponding to 2 44 The above Z 44 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0171] In some embodiments, the sense strand comprises only nucleotide sequence I, and The antisense strand contains only nucleotide sequence II.

[0172] In some embodiments, the nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 601 The nucleotide difference between the cleotide sequence and / or the nucleotide sequence is one or less, and / or the nucleotide sequence There is a nucleotide difference of 1 between sequence II and the nucleotide sequence shown in sequence number 602. It is one or less.

[0173] In some embodiments, the nucleotide sequence II and sequence number 602 are shown The nucleotide difference between the nucleotide sequence and the nucleotide sequence is Z 44 Including the difference in position, Z 44 but Selected from U, C, or G. In some embodiments, the nucleotide difference is Z 44 This is the difference at the position of Z 44 is selected from U, C, or G. Several implementation forms In state, Z 43 is, Z 44 It is a complementary nucleotide. The above nucleotide differences The siRNA possesses high target mRNA repression ability and contains these nucleotide differences. siRNA is also within the scope of protection of this disclosure.

[0174] In some embodiments, the nucleotide sequence I and the nucleotide sequence II are They are basically inversely complementary, substantially inversely complementary, or completely inversely complementary.

[0175] In some embodiments, nucleotide sequence I is the nucleotide shown in SEQ ID NO: 603 The rheotide sequence is the nucleotide sequence II shown in SEQ ID NO: 604. It is an array. 5'-CUUGCUCUGAAGUAGAAAZ 43 -3'(Sequence ID 603), 5'-Z 44 UUUCUACUUCAGAGCAAG-3' (Sequence ID 604) However, the aforementioned Z 44 Z is the first nucleotide at the 5' end of the antisense strand. 44 is selected from A, U, G, or C, and Z 43 is, Z 44 It is a complementary nucleotide. In some embodiments, Z 43 is U, Z 44 It is A.

[0176] The sense chain and the antisense chain may have the same or different lengths, and the length of the sense chain may be It consists of 19-23 nucleotides, and the antisense strand is 19-26 nucleotides long. .

[0177] In some embodiments, the sense strand further comprises nucleotide sequence III. The antisense strand further comprises nucleotide sequence IV and nucleotide sequence III The nucleotide sequence IV has a length of 1 to 4 nucleotides, and the nucleotide sequence IV has a length of 1 to 4 nucleotides. Sequence III and nucleotide sequence IV are of equal length and substantially inversely complementary or complete. It is inversely complementary to the above, and the nucleotide sequence III is at the 5' end of the nucleotide sequence I. The nucleotide sequence IV is bound to the 3' end of the nucleotide sequence II. In some embodiments, the nucleotide sequence IV and the second nucleotide sequence The sequence is substantially inversely complementary or completely inversely complementary, and is related to the second nucleotide sequence. This is adjacent to the 5' end of the nucleotide sequence shown in SEQ ID NO: 601 in the target mRNA. This refers to a nucleotide sequence whose length is equal to that of nucleotide sequence IV.

[0178] In some embodiments, the nucleotide sequence III follows the 5'-3' direction. Nucleotide sequence IV and nucleotide sequence I are both 1 nucleotide in length, and nucleotide sequence I The base of nucleotide II is G, and the base of nucleotide sequence IV is C. In this case, the sense strand and The antisense strand length ratio is 20 / 20, or nucleotide sequences III and IV are Each has a length of 2 nucleotides, and from the 5' end to the 3' end, the nucleoti The base sequence of nucleotide sequence III is GG, and the base sequence of nucleotide sequence IV is CC. At this time, the length ratio of the sense chain to the antisense chain is 21 / 21, or nucleoty Sequences III and IV are both 3 nucleotides long, and from the 5' end to the 3' end... Towards the end, the base sequence of nucleotide sequence III is UGG, and the base sequence of nucleotide sequence IV is The base sequence is CCA, and in this case, the length ratio of the sense strand to the antisense strand is 22 / 22. Alternatively, nucleotide sequences III and IV are both 4 nucleotides long. From the 5' end to the 3' end, the base sequence of nucleotide sequence III is GUGG. Furthermore, the base sequence of nucleotide sequence IV is CCAC, and in this case, the sense strand and antisequence The nucleotide chain length ratio is 23 / 23. In some embodiments, the nucleotide arrangement Column III and nucleotide sequence IV are 2 nucleotides long, from the 5' end to the 3' end. Towards the end, the base sequence of nucleotide sequence III is GG, and nucleotide sequence IV The base sequence is CC, and in this case, the length ratio of the sense strand to the antisense strand is 21 / 21. be.

[0179] In some embodiments, nucleotide sequences III and nucleotide IV are complete Since they are inversely complementary, given the bases of nucleotide sequence III, nucleotide The bases of sequence IV are also determined.

[0180] <The 12th siRNA> According to this disclosure, the siRNA may be a twelfth siRNA.

[0181] The 12th siRNA comprises a sense strand and an antisense strand. Each nucleotide is independently modified or unmodified, and the sense The strand contains nucleotide sequence I, and the antisense strand contains nucleotide sequence II, The nucleotide sequence I and the nucleotide sequence II are, at least in part, inversely complementary. This chain region is formed by the nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 661. A row is defined as a sequence of nucleotides of equal length with a nucleotide difference of 3 or less, as described in the nucleotide sequence II. The nucleotide sequence shown in sequence number 662 is of equal length, and the nucleotide difference is There are three or fewer. 5'-CUUCUUUGCCAUCAAAGAZ 45 -3'(Sequence ID 661), 5'-Z 46 UCUUUGAUGGCAAAGAAG-3' (Sequence ID 662) However, Z 45 is U, Z46 is A, and in the nucleotide sequence I, position Z 45 Nucleotide Z corresponding to 47 It includes, and the nucleotide sequence II has a position Z4 Nucleotide Z corresponding to 6 48 The above Z 48 This is the 5' end of the antisense chain. It is the first nucleotide at the end.

[0182] In some embodiments, the sense strand comprises only nucleotide sequence I, and The antisense strand contains only nucleotide sequence II.

[0183] In some embodiments, the nucleotide sequence I and the nucleotide sequence shown in SEQ ID NO: 661 The nucleotide difference between the cleotide sequence and / or the nucleotide sequence is one or less, and / or the nucleotide sequence There is a nucleotide difference of 1 between sequence II and the nucleotide sequence shown in sequence number 662. It is one or less.

[0184] In some embodiments, the nucleotide sequence II and sequence number 662 are shown The nucleotide difference between the nucleotide sequence and the nucleotide sequence is Z 48 Including the difference in position, Z 48 but Selected from U, C, or G. In some embodiments, the nucleotide difference is Z 48 This is the difference at the position of Z 48 is selected from U, C, or G. Several implementation forms In state, Z 47 is, Z 48 It is a complementary nucleotide. The above nucleotide differences The siRNA possesses high target mRNA repression ability and contains these nucleotide differences. siRNA is also within the scope of protection of this disclosure.

[0185] In some embodiments, the nucleotide sequence I and the nucleotide sequence II are They are basically inversely complementary, substantially inversely complementary, or completely inversely complementary.

[0186] In some embodiments, nucleotide sequence I is the nucleotide shown in SEQ ID NO: 663 The rheotide sequence is the nucleotide sequence II shown in SEQ ID NO: 664. It is an array. 5'-CUUCUUUGCCAUCAAAGAZ 47 -3'(Sequence ID 663), 5'-Z 48 UCUUUGAUGGCAAAGAAG-3' (Sequence ID 664) However, the aforementioned Z 48 Z is the first nucleotide at the 5' end of the antisense strand. 48 is selected from A, U, G, or C, and Z 47 is, Z 48 It is a complementary nucleotide. In some embodiments, Z 47 is U, Z 48 It is A.

[0187] The sense chain and the antisense chain may have the same or different lengths, and the length of the sense chain may be It consists of 19-23 nucleotides, and the antisense strand is 19-26 nucleotides long. .

[0188] In some embodiments, the sense strand further comprises nucleotide sequence III. The antisense strand further comprises nucleotide sequence IV and nucleotide sequence III The nucleotide sequence IV has a length of 1 to 4 nucleotides, and the nucleotide sequence IV has a length of 1 to 4 nucleotides. Sequence III and nucleotide sequence IV are of equal length and substantially inversely complementary or complete. It is inversely complementary to the above, and the nucleotide sequence III is at the 5' end of the nucleotide sequence I. The nucleotide sequence IV is bound to the 3' end of the nucleotide sequence II. In some embodiments, the nucleotide sequence IV and the second nucleotide sequence The sequence is substantially inversely complementary or completely inversely complementary, and is related to the second nucleotide sequence. This is adjacent to the 5' end of the nucleotide sequence shown in SEQ ID NO: 661 in the target mRNA. This refers to a nucleotide sequence whose length is equal to that of nucleotide sequence IV.

[0189] In some embodiments, the nucleotide sequence III follows the 5'-3' direction. Nucleotide sequence IV and nucleotide sequence I are both 1 nucleotide in length, and nucleotide sequence I The base of nucleotide II is U, and the base of nucleotide sequence IV is A. In this case, the sense strand and The antisense strand length ratio is 20 / 20, or nucleotide sequences III and IV are Each has a length of 2 nucleotides, and from the 5' end to the 3' end, the nucleoti The base sequence of sequence III is AU, and the base sequence of nucleotide sequence IV is AU. At this time, the length ratio of the sense chain to the antisense chain is 21 / 21, or nucleoty Sequences III and IV are both 3 nucleotides long, and from the 5' end to the 3' end... Towards the right, the base sequence of nucleotide sequence III is UAU, and the base sequence of nucleotide sequence IV is The base sequence is AUA, and in this case, the length ratio of the sense strand to the antisense strand is 22 / 22. Alternatively, nucleotide sequences III and IV are both 4 nucleotides long. From the 5' end to the 3' end, the base sequence of nucleotide sequence III is CUAU. Furthermore, the base sequence of nucleotide sequence IV is AUAG, and in this case, the sense strand and antisequence The nucleotide chain length ratio is 23 / 23. In some embodiments, the nucleotide arrangement Column III and nucleotide sequence IV are 2 nucleotides long, from the 5' end to the 3' end. Towards the end, the base sequence of nucleotide sequence III is AU, and nucleotide sequence IV The base sequence is AU, and in this case the length ratio of the sense strand to the antisense strand is 21 / 21. be.

[0190] In some embodiments, nucleotide sequences III and nucleotide IV are complete Since they are inversely complementary, given the bases of nucleotide sequence III, nucleotide The bases of sequence IV are also determined.

[0191] Below are nucleotide sequence V, nucleic acid sequence, nucleotide modification in siRNA, and The description of the modified sequence is applicable to any one of the 1st to 12th siRNAs listed above. In other words, unless otherwise specified, the following descriptions of siRNA refer to the first siRNA, the second siRNA, and so on. siRNA, 3rd siRNA, 4th siRNA, 5th siRNA, 6th siRNA NA, 7th siRNA, 8th siRNA, 9th siRNA, 10th siRNA, This should be considered a description of the 11th siRNA and the 12th siRNA, one by one. Yes. For example, if a specific siRNA is not specified, it says, "The siRNA is a nucleotide." "Further containing siRNA sequence V" means "the first siRNA, the second siRNA, and the third siRNA RNA, 4th siRNA, 5th siRNA, 6th siRNA, 7th siRNA, The 8th siRNA, the 9th siRNA, the 10th siRNA, the 11th siRNA or the This means that the 12 siRNAs further contain nucleotide sequence V.

[0192] In some embodiments, the antisense strand further comprises nucleotide sequence V Furthermore, the nucleotide sequence V has a length of 1 to 3 nucleotides, and the 3 antisense strands It is attached to the end and forms the 3' overhang end of the antisense chain. As a result, the sense strand-to-antisense strand length ratio of the siRNA provided in this disclosure is 19 / 20, 19 / 21, 19 / 22, 20 / 21, 20 / 22, 20 / 23, 21 / 22, 21 / 23, 21 / 24, 22 / 23, 22 / 24, 22 / 25, 23 / 24, 23 / 2 It may be 5 or 23 / 26. In some embodiments, the nucleotide sequence V has a length of 2 nucleotides, thereby the siRNA provided by this disclosure Even if the length ratio of the sense strand to the antisense strand is 19 / 21, 21 / 23, or 23 / 25 good.

[0193] Each nucleotide in the nucleotide sequence V may be any nucleotide. Furthermore, in order to facilitate synthesis and save on synthesis costs, the nucleotide sequence V is divided into two consecutive sequences. 1 thyminedeoxyribonucleotide (dTdT) or 2 consecutive uracilribonucleonucleotides It is a rheotide (UU), or the affinity between the antisense strand of siRNA and the target mRNA. To enhance its power, nucleotide sequence V is the nucleo at the corresponding position of the target mRNA. It is complementary to siRNA. Therefore, in some embodiments, the siRNA of this disclosure The ratio of the sense strand length to the antisense strand length is 19 / 21 or 21 / 23, and in this case, The disclosed siRNA has superior target mRNA silencing activity.

[0194] The nucleotide at the corresponding position of the target mRNA is the nucleo of the target mRNA at the 5' end. This refers to a nucleotide or nucleotide sequence adjacent to the nucleotide sequence of the target mRNA. The rheotide sequence is substantially or completely inversely complementary to nucleotide sequence II. It is either a creotide sequence or composed of nucleotide sequence II and nucleotide sequence IV. Nucleotide sequences that are substantially or completely inversely complementary to the nucleotide sequence being treated That is the case.

[0195] In some embodiments, the first siRNA is subjected to the siRNA's sensor The chain contains the nucleotide sequence shown in Sequence ID No. 5, and is the antisense of the siRNA. The chain contains the nucleotide sequence shown in Sequence ID No. 6. 5'-GAGAUGAAGUUCAAGAAUZ3-3'(Sequence ID 5), 5'-Z4AUUCUUGAACUUCAUCUCAA-3'(Sequence ID 6)

[0196] Alternatively, the sense strand of the siRNA may contain the nucleotide sequence shown in Sequence ID No. 7. The antisense strand includes the nucleotide sequence shown in Sequence ID No. 8. 5'-UUGAGAUGAAGUUCAAGAAUZ3-3'(Sequence ID 7), 5'-Z4AUUCUUGAACUUCAUCUCAAUG-3'(Sequence ID 8) However, the aforementioned Z4 is the first nucleotide at the 5' end of the antisense strand, and Z4 Z3 is selected from A, U, G, or C, and Z3 is a nucleotide complementary to Z4.

[0197] In some embodiments, the second siRNA is subjected to the siRNA's saturation. The chain contains the nucleotide sequence shown in Sequence ID No. 65, and the antisensor of the siRNA. The chain contains the nucleotide sequence shown in Sequence ID No. 66. 5'-CAUAACUGGAAUUUGUAAZ7-3'(SEQ ID NO: 65), 5'-Z8UUACAAAUUCCAGUUAUGUU-3'(Sequence ID 66)

[0198] Alternatively, the sense strand of the siRNA contains the nucleotide sequence shown in Sequence ID No. 67. Furthermore, the antisense strand of the siRNA contains the nucleotide sequence shown in SEQ ID NO: 68. nothing. 5'-AACAUAACUGGAAUUUGUAAZ7-3'(Sequence ID 67), 5'-Z8UUACAAAUUCCAGUUAUGUUAC-3'(Sequence ID 68) However, the aforementioned Z8 is the first nucleotide at the 5' end of the antisense strand, and Z8 Z7 is selected from A, U, G, or C, and Z7 is a nucleotide complementary to Z8.

[0199] In some embodiments, the third siRNA is used to obtain the siRNA. The siRNA chain comprises the nucleotide sequence shown in Sequence ID No. 125, and the antiseptic of the siRNA. The nucleotide chain contains the nucleotide sequence shown in SEQ ID NO: 126. 5'-CAUUAUCACAAUUGAGGAZ 11 -3'(Sequence ID 125), 5'-Z 12 UCCUCAAUUGUGAUAAUGGC-3' (Sequence ID 126)

[0200] Alternatively, the sense strand of the siRNA may have the nucleotide sequence shown in Sequence ID No. 127 The antisense strand of the siRNA includes the nucleotide sequence shown in SEQ ID NO: 128. Includes. 5'-GCCAUUAUCACAAUUGAGGAZ 11 -3'(Sequence ID 127), 5'-Z 12 UCCUCAAUUGUGAUAAUGGCUG-3'(Sequence ID 128) ) However, the aforementioned Z 12 Z is the first nucleotide at the 5' end of the antisense strand. 12 is selected from A, U, G, or C, and Z 11 is, Z 12 It is a complementary nucleotide. ru.

[0201] In some embodiments, the fourth siRNA is used to obtain the siRNA. The siRNA chain comprises the nucleotide sequence shown in Sequence ID No. 185, and the antiseptic of the siRNA. The nucleotide chain contains the nucleotide sequence shown in SEQ ID NO: 186. 5'-GGAUCUCUCUCAGAGUAUZ 15 -3'(Sequence ID 185), 5'-Z 16 AUACUCUGAGAGAGAUCCUG-3' (Sequence ID 186)

[0202] Alternatively, the sense strand of the siRNA may have the nucleotide sequence shown in Sequence ID No. 187 The antisense strand of the siRNA includes the nucleotide sequence shown in SEQ ID NO: 188. Includes. 5'-CAGGAUCUCUCUCAGAGUAUZ 15 -3'(Sequence ID 187), 5'-Z 16 AUACUCUGAGAGAGAUCCUGGG-3'(Sequence ID 188) ) However, the aforementioned Z 16 Z is the first nucleotide at the 5' end of the antisense strand. 16 is selected from A, U, G, or C, and Z 15 is, Z16 It is a complementary nucleotide. ru.

[0203] In some embodiments, the siRNA is used with respect to the fifth siRNA. The siRNA chain comprises the nucleotide sequence shown in Sequence ID No. 245, and the antiseptic of the siRNA. The nucleotide chain contains the nucleotide sequence shown in SEQ ID NO: 246. 5'-ACAUGGACAACUGCUAUAZ 19 -3'(Sequence ID 245), 5'-Z 20 UAUAGCAGUUGUCCAUGUGG-3' (Sequence ID 246)

[0204] Alternatively, the sense strand of the siRNA may have the nucleotide sequence shown in Sequence ID No. 247 The antisense strand of the siRNA includes the nucleotide sequence shown in SEQ ID NO: 248. Includes. 5'-CCACAUGGACAACUGCUAUAZ 19 -3'(Sequence ID 247), 5'-Z 20 UAUAGCAGUUGUCCAUGUGGAA-3'(Sequence No. 248) ) However, the aforementioned Z 20 Z is the first nucleotide at the 5' end of the antisense strand. 20 is selected from A, U, G, or C, and Z 19 is, Z 20 It is a complementary nucleotide. ru.

[0205] In some embodiments, the siRNA is used with respect to the sixth siRNA. The siRNA chain comprises the nucleotide sequence shown in Sequence ID No. 305, and the antiseptic of the siRNA. The nucleotide chain contains the nucleotide sequence shown in SEQ ID NO: 306. 5'-UAGCAAGCUCUCAGUAUCZ23 -3'(Sequence ID 305), 5'-Z 24 GAUACUGAGAGCUUGCUAGG-3' (SEQ ID NO: 306)

[0206] Alternatively, the sense strand of the siRNA may have the nucleotide sequence shown in SEQ ID NO: 307 The antisense strand of the siRNA includes the nucleotide sequence shown in SEQ ID NO: 308. Includes. 5'-CCUAGCAAGCUCUCAGUAUCZ 23 -3'(Sequence ID 307), 5'-Z 24 GAUACUGAGAGCUUGCUAGGCA-3'(Sequence No. 308) ) However, the aforementioned Z 24 Z is the first nucleotide at the 5' end of the antisense strand. 24 is selected from A, U, G, or C, and Z 23 is, Z 24 It is a complementary nucleotide. ru.

[0207] In some embodiments, the seventh siRNA is used to obtain the siRNA. The siRNA chain comprises the nucleotide sequence shown in Sequence ID No. 365, and the antiseptic of the siRNA. The nucleotide chain contains the nucleotide sequence shown in SEQ ID NO: 366. 5'-AUAAGGUUACUUGUGUUGZ 27 -3'(Sequence ID 365), 5'-Z 28 CAACACAAGUAACCUUAUCC-3' (Sequence ID 366)

[0208] Alternatively, the sense strand of the siRNA may have the nucleotide sequence shown in SEQ ID NO: 367 The antisense strand includes the nucleotide sequence shown in SEQ ID NO: 368. 5'-GGAUAAGGUUACUUGUGUUGZ27 -3'(Sequence ID 367), 5'-Z 28 CAACACAAGUAACCUUAUCCUU-3'(Sequence ID 368) ) However, the aforementioned Z 28 Z is the first nucleotide at the 5' end of the antisense strand. 28 is selected from A, U, G, or C, and Z 27 is, Z 28 It is a complementary nucleotide. ru.

[0209] In some embodiments, the siRNA of the eighth siRNA is used. The siRNA chain comprises the nucleotide sequence shown in Sequence ID No. 425, and the antiseptic of the siRNA. The nucleotide chain contains the nucleotide sequence shown in SEQ ID NO: 426. 5'-GAAAAUCACCUAUGAAGAZ 31 -3'(Sequence ID 425), 5'-Z 32 UCUUCAUAGGUGAUUUUCAC-3' (Sequence ID 426)

[0210] Alternatively, the sense strand of the siRNA may have the nucleotide sequence shown in Sequence ID No. 427 The antisense strand of the siRNA includes the nucleotide sequence shown in SEQ ID NO: 428. Includes. 5'-GUGAAAAUCACCUAUGAAGAZ 31 -3'(Sequence ID 427), 5'-Z 32 UCUUCAUAGGUGAUUUUCACCC-3'(Sequence ID 428) ) However, the aforementioned Z 32 Z is the first nucleotide at the 5' end of the antisense strand. 32 is selected from A, U, G, or C, and Z 31 is, Z 32 It is a complementary nucleotide. ru.

[0211] In some embodiments, the siRNA of the 9th siRNA is used. The siRNA chain comprises the nucleotide sequence shown in Sequence ID No. 485, and the antiseptic of the siRNA. The nucleotide chain contains the nucleotide sequence shown in SEQ ID NO: 486. 5'-GAUGCUAUAAAGAACAACZ 35 -3'(Sequence ID 485), 5'-Z 36 GUUGUUCUUUAUAGCAUCCU-3' (Sequence ID 486)

[0212] Alternatively, the sense strand of the siRNA may have the nucleotide sequence shown in Sequence ID No. 487 The antisense strand of the siRNA includes the nucleotide sequence shown in SEQ ID NO: 488. Includes. 5'-AGGAUGCUAUAAAGAACAACZ 35 -3'(Sequence ID 487), 5'-Z 36 GUUGUUCUUUAUAGCAUCCUCA-3'(Sequence ID 488) ) However, the aforementioned Z 36 Z is the first nucleotide at the 5' end of the antisense strand. 36 is selected from A, U, G, or C, and Z 35 is, Z 36 It is a complementary nucleotide. ru.

[0213] In some embodiments, the siRNA is used with respect to the 10th siRNA. The siRNA chain comprises the nucleotide sequence shown in SEQ ID NO: 545, and the antinucleotide of the siRNA. The sense strand contains the nucleotide sequence shown in SEQ ID NO: 546. 5'-GAACAACUCCUUUUAUGGZ 39 -3'(Sequence ID 545), 5'-Z 40 CCAUAAAAGGAGUUGUUCUU-3' (Sequence ID 546)

[0214] Alternatively, the sense strand of the siRNA may have the nucleotide sequence shown in SEQ ID NO: 547 The antisense strand of the siRNA includes the nucleotide sequence shown in SEQ ID NO: 548. Includes. 5'-AAGAACAACUCCUUUUAUGGZ 39 -3'(Sequence ID 547), 5'-Z 40 CCAUAAAAGGAGUUGUUCUUUA-3'(Sequence ID 548) ) However, the aforementioned Z 40 Z is the first nucleotide at the 5' end of the antisense strand. 40 is selected from A, U, G, or C, and Z 39 is, Z 40 It is a complementary nucleotide. ru.

[0215] In some embodiments, the siRNA is used with respect to the 11th siRNA. The siRNA chain comprises the nucleotide sequence shown in SEQ ID NO: 605, and the antinucleotide of the siRNA. The sense strand contains the nucleotide sequence shown in SEQ ID NO: 606. 5'-CUUGCUCUGAAGUAGAAAZ 43 -3'(Sequence ID 605), 5'-Z 44 UUUCUACUUCAGAGCAAGCC-3' (Sequence ID 606)

[0216] Alternatively, the sense strand of the siRNA may have the nucleotide sequence shown in SEQ ID NO: 607 The antisense strand of the siRNA includes the nucleotide sequence shown in SEQ ID NO: 608. Includes. 5'-GGCUUGCUCUGAAGUAGAAAZ43 -3'(Sequence ID 607), 5'-Z 44 UUUCUACUUCAGAGCAAGCCAC-3'(Sequence ID 608) ) However, the aforementioned Z 44 Z is the first nucleotide at the 5' end of the antisense strand. 44 is selected from A, U, G, or C, and Z 43 is, Z 44 It is a complementary nucleotide. ru.

[0217] In some embodiments, the siRNA is used with respect to the 12th siRNA. The siRNA chain comprises the nucleotide sequence shown in SEQ ID NO: 665, and the antinucleotide of the siRNA. The sense strand contains the nucleotide sequence shown in SEQ ID NO: 666. 5'-CUUCUUUGCCAUCAAAGAZ 47 -3'(Sequence ID 665), 5'-Z 48 UCUUUGAUGGCAAAGAAGAU-3' (Sequence ID 666)

[0218] Alternatively, the sense strand of the siRNA may have the nucleotide sequence shown in SEQ ID NO: 667 The antisense strand of the siRNA includes the nucleotide sequence shown in SEQ ID NO: 668. Includes. 5'-AUCUUCUUUGCCAUCAAAGAZ 47 -3'(Sequence ID 667), 5'-Z 48 UCUUUGAUGGCAAAGAAGAUAG-3'(Sequence ID 668) ) However, the aforementioned Z 48 Z is the first nucleotide at the 5' end of the antisense strand. 48 is selected from A, U, G, or C, and Z 47 is, Z 48 It is a complementary nucleotide. ru.

[0219] In some embodiments, the siRNAs described in this disclosure are as shown in Tables 1a to 1l. The listed siXOa1, siXOa2, siXOb1, siXOb2, siXOc1 , siXOc2, siXOd1, siXOd2, siXOe1, siXOe2, siXO f1, siXOf2, siXOg1, siXOg2, siXOh1, siXOh2, si XOi1, siXOi2, siXOj1, siXOj2, siXOk1, siXOk2, It is either siXOl1 or siXOl2.

[0220] As mentioned above, each nucleotide in the siRNA of this disclosure is independently modified or This is an unmodified nucleotide. In some embodiments, each of the siRNAs of this disclosure All nucleotides are unmodified nucleotides. In some embodiments, Some or all of the nucleotides in the disclosed siRNA are modified nucleotides, These modifications on the creotide group allow the siRNA complex of this disclosure to suppress XO gene expression. It does not significantly weaken or cause loss of the controlling function.

[0221] In some embodiments, the siRNAs of this disclosure have at least one modified nucleo Contains nucleotides. In the context of this specification, the term "modified nucleotide" means a nucleotide. A nucleotide or nucleotide in which the hydroxyl group at the 2' position of the ribose group of a rheotide is substituted with another group. This refers to a nucleotide having a cleotide analog or a modified base. Cleotides clearly weaken or eliminate the gene expression repressive function of siRNA. It's not like that. For example, JK Watts, GF Deleavey, and MJ Damha,Chemically modified siRNA: t ools and applications. Drug Discov Today Select the modified nucleotides disclosed in 2008, 13(19-20): 842-55. You may do so.

[0222] In some embodiments, the sense strand of the siRNA provided herein or the aforementioned The antisense strand has at least one modified nucleotide, and / or It is a phosphate ester group in which at least one phosphate ester group has a modifying group. Alternatively, at least one single-stranded phosphate group in the sense chain and the antisense chain. - At least a portion of the phosphate ester groups and / or ribose groups in the sugar skeleton have modifying groups. It is a ribose group having a phosphate ester group and / or a modifying group.

[0223] In some embodiments, the sense chain and / or the antisense chain Cleotides are all modified nucleotides. In some embodiments, as disclosed herein Each nucleotide in the sense strand and antisense strand of the provided siRNA is They are either fluoromodified nucleotides or non-fluoromodified nucleotides.

[0224] Surprisingly, the inventors of this disclosure have found that the siRNA described herein can be used in animal experiments. In this case, a high degree of balance is achieved between plasma stability and gene silencing efficiency. I found it.

[0225] In some embodiments, the fluoromodified nucleotide is nucleotide sequence I And located in nucleotide sequence II, from the 5' end toward the 3' end, the nucleotide At least the nucleotides at positions 7, 8, and 9 of sequence I are fluoromodified nucleotides. From the 5' end toward the 3' end, at least the 2nd, 6th, and 1st nucleotides of the nucleotide sequence II The nucleotides at positions 4 and 16 are fluoromodified nucleotides.

[0226] In some embodiments, the fluoromodified nucleotide is nucleotide sequence I It is located in nucleotide sequence II, and is a fluoromodified nucleus in nucleotide sequence I. There are 5 or fewer ocides, and from the 5' end to the 3' end, the nucleotide sequence I At least the nucleotides at positions 7, 8, and 9 are fluoromodified nucleotides, and the nucleotides The fluoromodified nucleotides in rheotide sequence II are 7 or fewer, and the nucleo At least the nucleotides at positions 2, 6, 14, and 16 of sequence II are fluoromodified nucleos It's Chido.

[0227] In some embodiments, the sense strand odor is located from the 5' end to the 3' end. Then, the nucleotides at positions 7, 8, and 9 or 5, 7, 8, and 9 of the nucleotide sequence I are It is a ruoro-modified nucleotide, and the remaining nucleotides in the sense strand are non-fu It is a ruoro-modified nucleotide, and from the 5' end to the 3' end, the antisense strand In the above nucleotide sequence II, positions 2, 6, 14, 16 or 2, 6, 8, 9, 1 The nucleotides at positions 4 and 16 are fluoromodified nucleotides, and the antisense strand is The remaining nucleotides are un-fluoromodified nucleotides.

[0228] In the context of this specification, "fluoromodified nucleotide" means a nucleotide with ribochemical modification. The structure shown in formula (7) below has a hydroxyl group at the 2' position of the hydroxyl group substituted with fluorine. This refers to nucleotides that undergo ribodynamic modification. "Non-fluoromodified nucleotides" are nucleotides that undergo ribodynamic modification. A nucleotide in which the hydroxyl group at the 2' position of the nucleotide is replaced with a nonfluorine group, or a nucleotide Refers to an analog. In some embodiments, each non-fluoromodified nucleotide is a nucleus A nucleotide in which the hydroxyl group at the 2' position of the ribose group of a rheotide is replaced with a nonfluorine group. This is one of the nucleotide analogs that is selected independently.

[0229] These nucleotides, in which the hydroxyl group at the 2' position of the ribose group is replaced with a nonfluorine group, are These nucleotides are known to those skilled in the art, and are 2'-alkoxy modified nucleotides. 2'-substituted alkoxy modified nucleotides, 2'-alkyl modified nucleotides, 2'-substituted Alkyl-modified nucleotides, 2'-amino-modified nucleotides, 2'-substituted amino-modified nucleotides The rheotide may be one selected from 2'-deoxynucleotides.

[0230] In some embodiments, the 2'-alkoxy-modified nucleotide is shown in formula (8) It is a methoxy-modified nucleotide (2'-OMe). In some embodiments, 2'-substituted alkoxy modified nucleotides include, for example, the 2'-O-methionine shown in formula (9). It may also be a xyethyl-modified nucleotide (2'-MOE). In some embodiments... The 2'-amino modified nucleotide (2'-NH2) is shown in formula (10). In that embodiment, the 2'-deoxynucleotide (DNA) is represented by formula (11) .

[0231] [ka]

[0232] Nucleotide analogs are substances that can substitute for nucleotides in nucleic acids. adenine ribonucleotide, guanine ribonucleotide, cytosine ribonucleotide, This refers to a group whose structure differs from that of uracil ribonucleotide or thymine deoxyribonucleotide. In some embodiments, the nucleotide analog is an isonucleotide, a cross-linked nucleotide. rheotide (bridged nucleic acid, BNA) or acyclic nucleotide It's also fine to use "do".

[0233] BNA refers to a nucleotide that is constrained or cannot be accessed. BNA is a five-membered ring, six-membered ring. Includes a crosslinked structure having a ring or seven-membered ring with "fixed" C3'-endosugar puckering. That's fine. Usually, the bridge is introduced at the 2'- and 4'- positions of the ribose, resulting in 2',4'-BN. Provides A nucleotide. In some embodiments, BNA is represented by formula (12). LNA, ENA shown in equation (13), cET BNA shown in equation (14), etc. That's fine.

[0234] [ka]

[0235] Acyclic nucleotides are nucleotides in which the sugar ring of the nucleotide has been opened. In that embodiment, the acyclic nucleotide is an unlocked nucleic acid represented by formula (15). (UNA), or glycerol nucleic acid (GNA) represented by formula (16).

[0236] [ka] In formulas (15) and (16) above, R is H, OH, or alkoxy (O-alkoxy Selected from (Lu)

[0237] Isonucleotides are nucleotides in which the position of the base in the ribose ring changes. This refers to a compound. In some embodiments, the isonucleotide is of formula (17) or ( 18) The chemicals in which the base is transposed from the 1'-position to the 2'-position or 3'-position of the ribose ring. Mixed ingredients are also acceptable.

[0238] [ka] In the compounds of formulas (17) to (18) above, the base is A, U, G, C, or T, etc. R represents a nucleic acid base, and R is selected from H, OH, F, or the non-fluorinated group mentioned above.

[0239] In some embodiments, the nucleotide analog is an isonucleotide, LNA, It is one selected from ENA, cET, UNA, and GNA. In some embodiments Each non-fluoromodified nucleotide is a methoxy-modified nucleotide, In the pulse, the methoxy-modified nucleotide is defined as having a 2'-hydroxyl group of ribose. This refers to a nucleotide that has been substituted with a toxic molecule.

[0240] In this context, "fluoromodified nucleotide" and "2'-fluoromodified nucleotide" are used. "A nucleotide in which the 2'-hydroxyl group of the ribose group is replaced with fluorine" and "2'- "Nucleotides containing a fluororibose group" have the same meaning, and both are nucleotides. Compounds having the structure shown in formula (7), in which the 2'-hydroxyl group of is replaced with fluorine. This refers to "methoxy-modified nucleotides," "2'-methoxy-modified nucleotides," and "ribo." "nucleotides in which the 2'-hydroxyl group of the - group is replaced with methoxy" and "2'-methyl "Nucleotides containing a siribose group" have the same meaning, and both refer to nucleotides. The 2'-hydroxyl group of the bose group is substituted with methoxy, and the structure is as shown in formula (8). It refers to compounds.

[0241] In some embodiments, the siRNAs of this disclosure are siRNAs having the following modifications. That is, in the sense chain, from the 5' end toward the 3' end, the nucleus Nucleotides at positions 7, 8, and 9 or 5, 7, 8, and 9 of rheotide sequence I are fluoromodified. It is a nucleotide, and the remaining nucleotides in the sense strand are methoxy-modified It is a creotide, and in the antisense strand, the 2nd and 6th nucleotides of nucleotide sequence II , nucleotides at positions 14, 16 or 2, 6, 8, 9, 14, 16 are fluoromodified nucleotides It is a rheotide, and the remaining nucleotides in the antisense chain are methoxy-modified. It is a nucleotide.

[0242] In some embodiments, the siRNAs of this disclosure are siRNAs having the following modifications. That is, from the 5' end toward the 3' end, in the sense strand of the siRNA Nucleotides at positions 5, 7, 8 and 9 of nucleotide sequence I are fluoromodified nucleotides. The nucleotides at the remaining positions of the sense strand of the siRNA are methoxy-modified nucleotides. Furthermore, from the 5' end to the 3' end, the antisense strand of the siRNA The nucleotides at positions 2, 6, 8, 9, 14 and 16 of nucleotide sequence II are full Oro-modified nucleotides are nucleotides in the remaining positions of the antisense strand of the siRNA. These are methoxy-modified nucleotides, Alternatively, from the 5' end to the 3' end, the nucleus in the sense strand of the siRNA The nucleotides at positions 5, 7, 8, and 9 of the ocidal sequence I are fluoromodified nucleotides. The remaining nucleotides in the sense strand of the siRNA are methoxy-modified nucleotides. Furthermore, from the 5' end toward the 3' end, the nucleotide in the antisense strand of the siRNA Nucleotides at positions 2, 6, 14, and 16 of the creotide sequence II are fluoromodified nucleos It is a nucleotide, and the nucleotides at the remaining positions of the siRNA antisense strand are methoxy-modified. It is a creotide, Alternatively, from the 5' end to the 3' end, the nucleus in the sense strand of the siRNA The nucleotides at positions 7, 8, and 9 of the ocidal sequence I are fluoromodified nucleotides, The remaining nucleotides in the sense strand of the iRNA are methoxy-modified nucleotides, Furthermore, from the 5' end toward the 3' end, the nuclei in the antisense strand of the siRNA The nucleotides at positions 2, 6, 14, and 16 of the Otid sequence II are fluoromodified nucleotides. Therefore, the nucleotides at the remaining positions of the siRNA antisense strand are methoxy-modified nuclei. It's Ochido.

[0243] In some embodiments, the siRNAs provided herein are as shown in Tables 1a to 1l. The following are listed: siXOa1-M1, siXOa1-M2, siXOa1-M3, si XOa2-M1, siXOa2-M2, siXOa2-M3, siXOb1-M1, si XOb1-M2, siXOb1-M3, siXOb2-M1, siXOb2-M2, si XOb2-M3, siXOc1-M1, siXOc1-M2, siXOc1-M3, si XOc2-M1, siXOc2-M2, siXOc2-M3, siXOd1-M1, si XOd1-M2, siXOd1-M3, siXOd2-M1, siXOd2-M2, si XOd2-M3, siXOe1-M1, siXOe1-M2, siXOe1-M3, si XOe2-M1, siXOe2-M2, siXOe2-M3, siXOf1-M1, si XOf1-M2, siXOf1-M3, siXOf2-M1, siXOf2-M2, si XOf2-M3, siXOg1-M1, siXOg1-M2, siXOg1-M3, si XOg2-M1, siXOg2-M2, siXOg2-M3, siXOh1-M1, si XOh1-M2, siXOh1-M3, siXOh2-M1, siXOh2-M2, si XOh2-M3, siXOi1-M1, siXOi1-M2, siXOi1-M3, si XOi2-M1, siXOi2-M2, siXOi2-M3, siXOj1-M1, si XOj1-M2, siXOj1-M3, siXOj2-M1, siXOj2-M2, si XOj2-M3, siXOk1-M1, siXOk1-M2, siXOk1-M3, si XOk2-M1, siXOk2-M2, siXOk2-M3, siXOl1-M1, si XOl1-M2, siXOl1-M3, siXOl2-M1, siXOl2-M2 and s iXOl2-M3, any one of them.

[0244] The siRNA having the above modification is not only low in cost but also resistant to ribonucleases in the blood This makes it more difficult to cleave nucleic acids, thereby improving the stability of nucleic acids. This further enhances the property of resistance to rease hydrolysis. In addition, the above modified siRNA targets It exhibits high mRNA inhibitory activity.

[0245] In some embodiments, the sense strand and antinucleotide provided herein are used. In the sense chain, the number of phosphate ester groups in at least one single-chain phosphate-sugar skeleton is small. At least some of them are phosphate ester groups having modifying groups. In some embodiments , the phosphate ester group having a modifying group, the phosphate diester bond in the phosphate ester group It is a thiophosphate ester group in which at least one oxygen atom is replaced by a sulfur atom. In several embodiments, the phosphate ester group having the modifying group is represented by formula (1) It is a thiophosphate ester group with a specific structure.

[0246] [ka]

[0247] These modifications stabilize the double-stranded structure of siRNA, resulting in high base pairing specificity. And it can maintain a high level of affinity.

[0248] In some embodiments, the siRNA provided in this disclosure contains thioline The acid ester group is connected to the first nucleotide at any one end of the sense strand or antisense strand and 2 Between the second nucleotide and the second nucleotide at any end of the sense strand or antisense strand Selected from the group consisting of the relationship between the ocide and the third nucleotide, or any combination thereof. It exists bound to at least one of them. In some embodiments, thiophosphate ester The tel group is present at all of the above positions except the 5' end of the sense chain. In the embodiment, the thiophosphate ester group is located at all of the above positions except the 3' end of the sense chain. It exists bonded to the following. In some embodiments, the thiophosphate ester group is as follows: It is attached to at least one of the locations. Between the first and second nucleotides from the 5' end of the sense strand, Between the second and third nucleotides from the 5' end of the sense strand, Between the first and second nucleotides from the 3' end of the sense strand, Between the second and third nucleotides from the 3' end of the sense strand, The first nucleotide and the second nucleotide from the 5' end of the aforementioned antisense strand Between, The second and third nucleotides from the 5' end of the aforementioned antisense strand Between, The first nucleotide and the second nucleotide from the 3' end of the aforementioned antisense strand between, and The second and third nucleotides from the 3' end of the aforementioned antisense strand Between.

[0249] In some embodiments, the siRNAs provided herein are as shown in Tables 1a to 1l. The following are listed: siXOa1-M1S, siXOa1-M2S, siXOa1-M3S , siXOa2-M1S, siXOa2-M2S, siXOa2-M3S, siXOb1 -M1S, siXOb1-M2S, siXOb1-M3S, siXOb2-M1S, si XOb2-M2S, siXOb2-M3S, siXOc1-M1S, siXOc1-M2 S, siXOc1-M3S, siXOc2-M1S, siXOc2-M2S, siXOc 2-M3S, siXOd1-M1S, siXOd1-M2S, siXOd1-M3S, s iXOd2-M1S, siXOd2-M2S, siXOd2-M3S, siXOe1-M 1S, siXOe1-M2S, siXOe1-M3S, siXOe2-M1S, siXO e2-M2S, siXOe2-M3S, siXOf1-M1S, siXOf1-M2S, siXOf1-M3S, siXOf2-M1S, siXOf2-M2S, siXOf2- M3S, siXOg1-M1S, siXOg1-M2S, siXOg1-M3S, siX Og2-M1S, siXOg2-M2S, siXOg2-M3S, siXOh1-M1S , siXOh1-M2S, siXOh1-M3S, siXOh2-M1S, siXOh2 -M2S, siXOh2-M3S, XOi1-M1S, siXOi1-M2S, siXO i1-M3S, siXOi2-M1S, siXOi2-M2S, siXOi2-M3S, siXOj1-M1S, siXOj1-M2S, siXOj1-M3S, siXOj2- M1S, siXOj2-M2S, siXOj2-M3S, siXOk1-M1S, siX Ok1-M2S, siXOk1-M3S, siXOk2-M1S, siXOk2-M2S , siXOk2-M3S, siXOl1-M1S, siXOl1-M2S, siXOl1 -M3S, siXOl2-M1S, siXOl2-M2 and siXOl2-M3S It is one of the following.

[0250] In some embodiments, the 5' terminal nucleoty of the antisense strand of the siRNA The nucleotide is a 5'-phosphate nucleotide or a 5'-phosphate analog modified nucleotide.

[0251] The conventional 5'-phosphate nucleotide or 5'-phosphate analog modified nucleotide is, It is known to those skilled in the art that, for example, a 5'-phosphate nucleotide may have the following structure. .

[0252] [ka]

[0253] Also, for example, Anastasia Khvorova and Jonathan K. Watts,The chemical evolution of oligo nucleotide therapies of clinical utility . Nature Biotechnology,2017,35(3): 238~4 Section 8 discloses the following four types of 5'-phosphate analog modified nucleotides.

[0254] [ka] In the formula, R is selected from H, OH, methoxy, and fluorine, and Base represents a nucleic acid base, A Selected from U, C, G, or T.

[0255] In some embodiments, the 5'-phosphate nucleotide is represented by formula (2), 5 nucleotides containing '-phosphate modification, and 5'-phosphate analog modified nucleotides are The vinyl phosphate ester (5'-(E)-vinylphospho shown in formula (3) A nucleotide containing a nate (E-VP) modification, or a thio shown in formula (5). It is a phosphate-modified nucleotide.

[0256] In some embodiments, the siRNAs provided herein are as shown in Tables 1a to 1l. The following are listed: siXOa1-M1P1, siXOa1-M2P1, siXOa1-M 3P1, siXOa2-M1P1, siXOa2-M2P1, siXOa2-M3P1, siXOa1-M1SP1, siXOa1-M2SP1, siXOa1-M3SP1, s iXOa2-M1SP1, siXOa2-M2SP1, siXOa2-M3SP1, si XOb1-M1P1, siXOb1-M2P1, siXOb1-M3P1, siXOb2 -M1P1, siXOb2-M2P1, siXOb2-M3P1, siXOb1-M1S P1, siXOb1-M2SP1, siXOb1-M3SP1, siXOb2-M1SP 1, siXOb2-M2SP1, siXOb2-M3SP1, siXOc1-M1P1, siXOc1-M2P1, siXOc1-M3P1, siXOc2-M1P1, siXO c2-M2P1, siXOc2-M3P1, siXOc1-M1SP1, siXOc1- M2SP1, siXOc1-M3SP1, siXOc2-M1SP1, siXOc2-M 2SP1, siXOc2-M3SP1, siXOd1-M1P1, siXOd1-M2P 1, siXOd1-M3P1, siXOd2-M1P1, siXOd2-M2P1, si XOd2-M3P1, siXOd1-M1SP1, siXOd1-M2SP1, siXO d1-M3SP1, siXOd2-M1SP1, siXOd2-M2SP1, siXOd 2-M3SP1, siXOe1-M1P1, siXOe1-M2P1, siXOe1-M 3P1, siXOe2-M1P1, siXOe2-M2P1, siXOe2-M3P1, siXOe1-M1SP1, siXOe1-M2SP1, siXOe1-M3SP1, s iXOe2-M1SP1、siXOe2-M2SP1、siXOe2-M3SP1、si XOf1-M1P1、siXOf1-M2P1、siXOf1-M3P1、siXOf2 -M1P1、siXOf2-M2P1、siXOf2-M3P1、siXOf1-M1S P1、siXOf1-M2SP1、siXOf1-M3SP1、siXOf2-M1SP 1、siXOf2-M2SP1、siXOf2-M3SP1、siXOg1-M1P1、 siXOg1-M2P1、siXOg1-M3P1、siXOg2-M1P1、siXO g2-M2P1、siXOg2-M3P1、siXOg1-M1SP1、siXOg1- M2SP1、siXOg1-M3SP1、siXOg2-M1SP1、siXOg2-M 2SP1、siXOg2-M3SP1、siXOh1-M1P1、siXOh1-M2P 1、siXOh1-M3P1、siXOh2-M1P1、siXOh2-M2P1、si XOh2-M3P1、siXOh1-M1SP1、siXOh1-M2SP1、siXO h1-M3SP1、siXOh2-M1SP1、siXOh2-M2SP1、siXOh 2-M3SP1、XOi1-M1P1、siXOi1-M2P1、siXOi1-M3P 1、siXOi2-M1P1、siXOi2-M2P1、siXOi2-M3P1、si XOi1-M1SP1、siXOi1-M2SP1、siXOi1-M3SP1、siX Oi2-M1SP1、siXOi2-M2SP1、siXOi2-M3SP1、siXO j1-M1P1、siXOj1-M2P1、siXOj1-M3P1、siXOj2-M 1P1、siXOj2-M2P1、siXOj2-M3P1、siXOk1-M1P1、 siXOk1-M2P1, siXOk1-M3P1, siXOk2-M1P1, siXO k2-M2P1, siXOk2-M3P1, siXOl1-M1P1, siXOl1-M 2P1, siXOl1-M3P1, siXOl2-M1P1, siXOl2-M2P1, siXOl2-M3P1, siXOj1-M1SP1, siXOj1-M2SP1, si XOj1-M3SP1, siXOj2-M1SP1, siXOj2-M2SP1, siX Oj2-M3SP1, siXOk1-M1SP1, siXOk1-M2SP1, siXO k1-M3SP1, siXOk2-M1SP1, siXOk2-M2SP1, siXOk 2-M3SP1, siXOl1-M1SP1, siXOl1-M2SP1, siXOl1 -M3SP1, siXOl2-M1SP1, siXOl2-M2SP1 and siXOl2 -M3SP1 is one of the following:

[0257] The inventors of this disclosure claim that the siRNA provided herein is plasma and lysosomal stable. Not only is the sex significantly improved, but they also unexpectedly discovered high target mRNA suppression activity. Ta.

[0258] The siRNAs provided in this disclosure are prepared using conventional siRNA preparation methods in the art (e.g., For example, it can be obtained by a solid-phase synthesis method and a liquid-phase synthesis method. Here, solid-phase synthesis is A commercial customization service is already available. Nucleosids with corresponding modifiers. By using a mer, modified nucleotide groups are introduced into the siRNA described herein. A method for preparing nucleoside monomers having the corresponding modifications, and modified nucleos Methods for introducing a rheotide group into siRNA are also well known to those skilled in the art.

[0259] <Drug composition> This disclosure relates to a drug comprising the above-mentioned siRNA as an active ingredient and a pharmaceutically acceptable carrier. A composition is provided.

[0260] Even if the pharmaceutically acceptable carrier is a carrier commonly used in the field of siRNA administration, Often, for example, magnetic nanoparticles, for example (nanoparticles based on Fe3O4 or Fe2O3), carbon nanotubes (carbon nanotubes, mesoporous silicon n), calcium phosphate nanoparticles Polyethylenes (PEI) , polyamidoamine dendrimer (PAMAM) d endrimer, polylysine (poly(L-lysine, PLL), chitosan) (chitosan), 1,2-dioleoyl-3-trimethylammonium propane ( 1,2-dioleoyl-3-trimethylammonium-propane (, DOTAP), poly-D- or L-lactic acid / glycolic acid copolymer (poly(D&L-l Actic / glycolic acid copolymer, PLGA, poly(a) (poly(2-aminoethyl ethylene phosphate ester)) lene phosphate), PPEEA), and poly(N,N-dimethylaminoethyl) (poly(2-dimethylaminoethyl methacrylate)) This includes acrylate, PDMAEMA, and one or more derivatives thereof. These are not the only options.

[0261] With respect to the content of siRNA and pharmaceutically acceptable carrier in the aforementioned drug composition, There is no requirement for this, but the usual content of each component may be acceptable. In some embodiments, The weight ratio of siRNA to a pharmaceutically acceptable carrier may be 1:(1-500). In some embodiments, the above weight ratio is 1:(1~50).

[0262] In some embodiments, the drug composition contains other pharmaceutically acceptable additives. It may be included, and the additive is one of the various formulations or compounds commonly used in this field. There may be multiple types. For example, the other pharmaceutically acceptable additives may be a pH buffer, It may contain at least one of a protective agent and an osmotic pressure regulator.

[0263] The aforementioned pH buffer is trishydroxymethylaminomethane hydrochloride with a pH of 7.5 to 8.5. Buffer solution (tris(hydroxymethyl) aminomethane hyd rochloride buffer and / or phosphate buffer with a pH of 5.5 to 8.5 It may also be a phosphate buffer solution with a pH of 5.5 to 8.5.

[0264] The aforementioned protective agents are inositol, sorbitol, sucrose, trehalose, and mannose. The drug combination may be at least one of maltose, lactose, and glucose. Based on the total weight of the product, the content of the protective agent may be 0.01 to 30% by weight.

[0265] The osmotic pressure regulator may be sodium chloride and / or potassium chloride. The content of the osmotic pressure regulator is such that the osmotic pressure of the drug composition is 200 to 700 milliosmoles / kilogram. It is determined to be ram (mOsm / kg). With the desired osmotic pressure, a person skilled in the art will know the above The content of the osmotic pressure regulator can be easily determined.

[0266] In some embodiments, the drug composition may be a liquid formulation such as an injection solution. Alternatively, it may be prepared as a lyophilized powder injection, mixed with a liquid additive at the time of administration, and used as a liquid formulation. The liquid formulation described above can be administered by subcutaneous, intramuscular, or intravenous injection, but is not limited to these methods. It is administered to the lungs by spraying, or to other organs and tissues (e.g., the liver) through the lungs by spraying. It can also be administered to, but is not limited to, the following embodiments. The drug composition is used for intravenous administration.

[0267] In some embodiments, the drug composition may be in the form of a liposomal formulation. i. In some embodiments, pharmaceutically acceptable substances used in the liposome formulation The carrier is an amine-containing transfection compound (hereinafter also called an organic amine), and an auxiliary lipid. Contains organic ammonium compounds and / or PEGylated (polyethylene glycolated) lipids. Here, the organic ammonium compounds The cereal, auxiliary lipids, and PEGylated lipids are described in their entirety in CN103380113A (quoted above). Amine-containing transfection compounds or their pharmaceutically acceptable properties as described in the specification (to be incorporated into the specification) One or more selected from acceptable salts or derivatives, auxiliary lipids, and PEGylated lipids Multiple species are acceptable.

[0268] In some embodiments, the organic amine is as described in CN103380113A. The compound shown in formula (201) or a pharmaceutically acceptable salt thereof may be used.

[0269] [ka] During the ceremony, each X 101 or X 102 Each of these is independently O, S, NA, or CA, where A is water. Prime or C1-C 20 It is a hydrocarbon chain, Each Y 101 or Z 101 Each of these is independently C=O, C=S, S=O, CH-OH or It is SO2, Each R 101 , R 102 , R 103 , R 104 , R 105 , R 106 or R 107 That Each independently consists of hydrogen, a cyclic or acyclic, substituted or unsubstituted, branched or linear aliphatic group, and a ring. Formula or acyclic, substituted or unsubstituted, branched or linear heteroaliphatic groups, substituted or unsubstituted , branched or linear acyl group, substituted or unsubstituted, branched or linear aryl group, substituted or unsubstituted A substituted, branched or linear heteroaryl group, x is an integer from 1 to 10, n is an integer between 1 and 3, m is an integer between 0 and 20, and p is either 0 or 1, where, If m=p=0, then R 102 It is hydrogen, If at least one of n or m is 2, then R 103 And nitrogen in equation (201) , forming a structure shown in formula (202) or formula (203).

[0270] [ka] In the formula, g, e, and f are each independent integers from 1 to 6, and "HCC" represents a hydrocarbon chain. This represents each * N represents the nitrogen atom in equation (201).

[0271] In some embodiments, R 103 is a polyamine. In other embodiments, R 103 is a ketal. In some embodiments, R in formula (201) 10 1 and R 102 Each of these can be independently and optionally substituted or unsubstituted, branched or linear aluminum chains. The alkyl or alkenyl is a alkyl or alkenyl having 3 to about 20 carbon atoms. For example, 8 to about 18 carbon atoms and 0 to 4 double bonds, for example, 0 to 2 double bonds They have a common characteristic.

[0272] In some embodiments, when n and m are independently a value of 1 or 3, R 103 This may be any one of the following equations (204) to (213).

[0273] [ka] In equations (204) to (213), g, e, and f are each independent integers from 1 to 6. Each "HCC" represents a hydrocarbon chain, and each * is R 103 and nitrogen in equation (201) It shows the bonding points with atoms, and any * Each H at position corresponds to the nitrogen atom in equation (201) and It may be substituted for joining purposes.

[0274] The compound shown in formula (201) is prepared according to the description in CN103380113A. That's good too.

[0275] In some embodiments, the organic amine is an organic amine represented by formula (214). and / or an organic amine represented by formula (215).

[0276] [ka]

[0277] The aforementioned supplementary lipids are cholesterol, cholesterol analogs and / or cholesterol It is a derivative of , The PEGylated lipid is 1,2-dipalmitamide-sn-glycero-3-phosphatidi Ethanolamine-N-[methoxy(polyethylene glycol)]-2000(1,2 -dipalmitoyl-sn-glycero-3-phosphatidylet hanolamine-N-[methoxy(polyethylene glyco (l)-2000)

[0278] In some embodiments, the drug composition includes the organic amine and the auxiliary lipid. The molar ratio of the PEGylated lipids is (19.7~80):(19.7~80):(0.3 It is ~50), and for example, it could be (50~70):(20~40):(3~20). stomach.

[0279] In some embodiments, the siRNA of the present disclosure and the amine-containing transfects The drug composition particles formed by the reagent have an average diameter of approximately 30 nm to approximately 200 nm. Generally, the wavelength is approximately 40 nm to 135 nm, and more generally, the average of the liposome particles. The diameter is approximately 50nm to 120nm, 50nm to 100nm, and 60nm to 90nm. The wavelength is approximately 70 nm to 90 nm, and for example, the average diameter of the liposome particles is approximately 30,4 0, 50, 60, 70, 75, 80, 85, 90, 100, 110, 120, 130, 1 The wavelength is 40, 150, or 160 nm.

[0280] In some embodiments, the siRNA of the present disclosure and the amine-containing transfects In a drug composition formed by a reagent, siRNA and total lipids (e.g., organic amines) The weight ratio (weight / weight ratio) of the supplementary lipids and / or PEGylated lipids is approximately 1:1 to 1:5 0, approximately 1:1~1:30, approximately 1:3~1:20, approximately 1:4~1:18, approximately 1:5~ Approximately 1:17, approximately 1:5 to approximately 1:15, approximately 1:5 to approximately 1:12, approximately 1:6 to approximately 1:12 or The ratio is within the range of approximately 1:6 to approximately 1:10, for example, the weight ratio of siRNA to total lipids in this disclosure is Approximately 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13 , 1:14, 1:15, 1:16, 1:17, or 1:18.

[0281] In some embodiments, the drug composition is such that each component exists independently at the time of market release. In some embodiments, this disclosure may exist as a liquid formulation at the time of use. Drug composition formed from siRNA provided by and the above-mentioned pharmaceutically acceptable carrier. These may be prepared according to various known methods, and instead of conventional siRNA, the present disclosure The provided siRNA can be used. In some embodiments, the following method is used. It may be prepared in a different way.

[0282] The organic amine, auxiliary lipid, and PEGylated lipid are suspended in alcohol in the above molar ratio and mixed uniformly. Combine to obtain a lipid solution. The amount of alcohol is such that the total mass concentration of the obtained lipid solution is 2-25 The concentration is determined to be mg / mL, for example, 8-18 mg / mL. The alcohol is Pharmaceutically acceptable alcohols, such as ethanol, propylene glycol, and benzyl Alcohol, glycerin, polyethylene glycol 200, polyethylene glycol 30 Selected from alcohols that are liquid at around room temperature, such as polyethylene glycol 400. It may be one or more types, for example, ethanol.

[0283] The siRNA provided in this disclosure is dissolved in a buffer salt solution to obtain an aqueous siRNA solution. The concentration of the buffer salt solution is 0.05 to 0.5 M, but it may also be, for example, 0.1 to 0.2 M. Adjust the pH of the buffer salt solution to 4.0-5.5, or for example, 5.0-5.2. The amount of buffer salt solution should be such that the siRNA concentration is 0.6 mg / mL or less, for example, 0.2~ The concentration is determined to be 0.4 mg / mL. The buffer salt is soluble acetate, soluble citrate One or more salts selected from salts, for example, sodium acetate and / or potassium acetate. It's fine if it's "um".

[0284] After mixing the lipid solution and the siRNA aqueous solution, the resulting product was heated at 40-60°C to a minimum temperature. Also, culture for 2 minutes, for example, 5 to 30 minutes to obtain the cultured liposome formulation. Lipid solution and s The volume ratio of the iRNA aqueous solutions is 1:(2~5), and may be, for example, 1:4.

[0285] The cultured liposome formulation is concentrated or diluted, impurities are removed, and sterilization is performed according to the present disclosure. The drug composition to be supplied is obtained. Its physicochemical parameters are such that the pH is 6.5 to 8. The encapsulation efficiency is 80% or higher, the particle size is 40-200 nm, and the polydispersity index is 0.30. The following conditions apply, and the osmotic pressure is 250-400 mOsm / kg. For example, physicochemical parameters The filter has a pH of 7.2-7.6, an encapsulation efficiency of 90% or more, and a particle size of 60-100 nm. The polydispersity index may be 0.20 or less, and the osmotic pressure may be 300 to 400 mOsm / kg.

[0286] Here, concentration or dilution may be performed before impurity removal, after impurity removal, or simultaneously. Any of various conventional methods may be employed as the method for removing impurities. For example, using a tangential flow system and a hollow fiber column, ultrafiltration may be performed under conditions of 100KDa, and the ultrafiltration exchange solution may be a phosphate buffer (PBS) at pH 7.4. As the sterilization method, any of various conventional methods may be employed. For example, filtration through a 0.22 μm filter may be performed for sterilization.

[0287] <siRNA complex> The present disclosure provides an siRNA complex including the above siRNA and a complexing group complexed and bound to the siRNA. [[ID=at]]

[0288] Generally, the complexing group includes at least one targeting group and an optional linker that are pharmaceutically acceptable, and the siRNA, the linker, and the targeting group are sequentially bound. In some embodiments, the number of the targeting groups is 1 to 6. In some embodiments, the number of the targeting groups is 2 to 4. The siRNA molecule may be complexed with the complexing group non-covalently or covalently. For example, it may be covalently complexed with the complexing group. The complexing site between the siRNA and the complexing group may be at the 3'-end or 5'-end of the sense strand of the siRNA, at the 5'-end of the antisense strand, or in the internal sequence of the siRNA. In some embodiments, the complexing site between the siRNA and the complexing group is at the 3'-terminal of the sense strand of the siRNA.

[0289] ​​​​​​In some embodiments, the complex group is the phosphate group of the nucleotide, the 2'-position nucleotide It may be bonded to a droxy group or a base. In some embodiments, the composite group is It may also be bonded to the hydroxyl group at the 3' position, in which case the nucleotide-nucleotide bond is 2'-5'. They are linked by nitrate diester bonds. The complex group is attached to the end of the siRNA chain. In this case, it is usually bound to the phosphate group of a nucleotide and to the internal sequence of siRNA. They are usually bonded to a ribose sugar ring or a base. For various bonding methods, see Muthia. h Manoharan et.al. siRNA conjugates carr ying sequentially assembled trivalent N- acetylgalactosamine linked through nucleus osides elicit robust gene silencing in v ivo in hepatocytes. ACS Chemical biology See 2015, 10 (5): 1181~7.

[0290] In some embodiments, the relationship between the siRNA and the complex group is acid-unstable or reducible. They may be bound by chemical bonds, and in the acidic environment of cell endosomes, these chemical bonds The compound is broken down, and the siRNA can be released. The complex group is bound to the sense strand of the siRNA, and the complex has an effect on siRNA activity. It can be reduced as much as possible.

[0291] In some embodiments, the pharmaceutically acceptable target group is the siRNA administration field. Ligands commonly used in this, for example, the various types described in WO2009082607A2 It may also be a ligand, and its disclosure as a whole is incorporated herein by reference. .

[0292] In some embodiments, the pharmaceutically acceptable target group is cholesterol, bile duct. Lipophilic molecules such as juice acids, vitamins (e.g., tocopherol), and lipid molecules with different chain lengths, Polymers such as polyethylene glycol, polypeptides such as membrane-permeable peptides, and apters Mer, antibodies, quantum dots, lactose, polylactose, mannose, galactose Sugars such as N-acetylgalactosamine (GalNAc) and folic acid (folate) , asialoglycoprotein, asialoglycoprotein residue, lipoprotein (e.g., high-density lipoprotein) (e.g., low-density lipoproteins), glucagon, neurotransmitters (e.g., adrenaline), growth Factors, target molecules such as receptor ligands expressed in hepatocytes such as transferrin, or their It may be one or more ligands selected from those formed by the derivatives.

[0293] In some embodiments, each ligand is a ligand that can bind to a receptor on the cell surface. Selected independently from the ligand. In some embodiments, at least one ligand It is a ligand that can bind to receptors on the surface of hepatocytes. In some embodiments, At least one ligand is a ligand that can bind to receptors on the surface of mammalian liver cells. In some embodiments, at least one ligand is a receptor on the surface of human hepatocytes. It is a ligand that can bind to. In some embodiments, at least one ligand It is a ligand that can bind to the asialoglycoprotein receptor (ASGPR) on the surface of the liver. The types of these ligands are known to those skilled in the art, and their action is generally to target the surface of cells. It binds to specific receptors on the surface and mediates the delivery of ligand-bound siRNA to target cells. do.

[0294] In some embodiments, the pharmaceutically acceptable target group is the surface of mammalian hepatocytes. One of the ligans that binds to the asialocryprotein receptor (ASGPR) on the surface It may also be a compound. In some embodiments, each ligand is independently an asialocactose compound. Proteins, for example, asialoorosomucoid (A) It is either SOR or asialofetuin (ASF). In several embodiments, the ligand is a sugar or a derivative of a sugar.

[0295] In some embodiments, at least one ligand is a sugar. Morphologically, each ligand is a sugar. In some embodiments, at least Another ligand is a monosaccharide, polysaccharide, modified monosaccharide, modified polysaccharide, or sugar derivative. In the embodiment, at least one of the ligands may be a monosaccharide, disaccharide, or trisaccharide. In some embodiments, at least one ligand is a modified sugar. In this embodiment, each ligand is a modified sugar. Each ligand is independently a polysaccharide, modified polysaccharide, monosaccharide, modified monosaccharide, polysaccharide derivative or Selected from monosaccharide derivatives. In some embodiments, each or a few of the ligands At least one of them is glucose and its derivatives, mannan and its derivatives, galactose and Its derivatives, xylose and its derivatives, ribose and its derivatives, fucose and its derivatives Conductors, lactose and its derivatives, maltose and its derivatives, arabinose and its derivatives Selected from the group consisting of conductors, fructose and its derivatives, and sialic acid.

[0296] In some embodiments, each ligand is D-mannopyrano, L-mannopyrano Pyranose, D-arabinose, D-xylofuranose, L-xylofuranose, D- Glucose, L-glucose, D-galactose, L-galactose, α-D-mannofura North, β-D-mannofuranose, α-D-mannopyranose, β-D-mannopyrano -α-D-glucopyranose, β-D-glucopyranose, α-D-glucofurano S, β-D-glucofuranose, α-D-fructofuranose, α-D-fructopyrano - α-D-galactopyranose, β-D-galactopyranose, α-D-galactof Lanose, β-D-galactofuranose, glucosamine, sialic acid, galactosamine, N - Acetylgalactosamine, N-trifluoroacetylgalactosamine, N-propionine Lugalactosamine, Nn-butyrylgalactosamine, N-isobutyrylgalactosamine , 2-amino-3-O-[(R)-1-carboxyethyl]-2-deoxy-β-D-g Lucopyranose, 2-deoxy-2-methylamino-L-glucopyranose, 4,6-di Deoxy-4-formamido-2,3-di-O-methyl-D-mannopyrano, 2-de Oxy-2-sulfamino-D-glucopyranose, N-glycolyl-α-neuraminic acid , 5-thio-β-D-glucopyranose, methyl 2,3,4-tris-O-acetyl-1 -Thio-6-O-trityl-α-D-glucopyranoside, 4-Thio-β-D-galactopyranoside Lanose, ethyl 3,4,6,7-tetra-O-acetyl-2-deoxy-1,5-diethyl O-α-D-glucoheptopyranoside, 2,5-anhydro-D-alonitrile, ribo From D-ribose, D-4-thioribose, L-ribose, or L-4-thioribose They may be selected independently. Other alternative ligands include, for example, CN1053780 You may also refer to the description in 82A, which, by reference, incorporates the entirety of its disclosure into this Specified. Born.

[0297] In some embodiments, the pharmaceutically acceptable label in the siRNA complex The target group may be galactose or N-acetylgalactosamine, or galactose or The N-acetylgalactosamine molecule may be monovalent, divalent, trivalent, or tetravalent. The terms monovalent, divalent, trivalent, and tetravalent refer to the siRNA molecule and the galactose used as the target group, respectively. An siRNA complex formed from a complex group containing a molecule of or N-acetylgalactosamine The molar ratio of siRNA molecules to galactose or N-acetylgalactosamine molecules is It should be understood that this refers to a ratio of 1:1, 1:2, 1:3, or 1:4. In the application form, the pharmaceutically acceptable target group is N-acetylgalactosamine. In some embodiments, the siRNA described herein is N-acetylgalactosa When compounded with a complex group containing mine, the N-acetylgalactosamine molecule is trivalent or tetravalent. In some embodiments, the siRNA described herein is N-acetylgalactam When combined with a complex group containing tosamine, the N-acetylgalactosamine molecule is trivalent.

[0298] The target group may be bound to the siRNA molecule via a suitable linker, and those skilled in the art will know The appropriate linker can be selected depending on the specific type of target substrate. These linkers, For information on the types of target groups and methods of binding to siRNA, please refer to WO2015006740A2. You may refer to the disclosed content, which will be incorporated into this specification in whole by citation.

[0299] In some embodiments, when the target group is N-acetylgalactosamine, A suitable linker may have the structure shown in formula (301).

[0300] [ka] During the ceremony, k is an integer between 1 and 3. L A This is a chain-like portion containing an amide bond having the structure shown in formula (302), and each of the above L A It has one target group at each end and the L C Bonded to the part by ether bond It will be done.

[0301] [ka] L B This is a chain portion containing N-acylpyrrolidine having the structure shown in formula (303). Furthermore, the chain portion has a carbonyl group at one end, and the L C The parts are bonded by amide bonds. It has an oxygen group at the other end and is bonded to the siRNA by a phosphate ester bond.

[0302] [ka] L CThis refers to hydroxymethylaminomethane, dihydroxymethylaminomethane, or trihydroxymethylaminomethane. A divalent to tetravalent linker group based on droxymethylaminomethane, and the L C is an oxygen source Each of the Ls is bonded via ether through the child. A It is bonded to the part, and via nitrogen atoms, The L B It is joined to the part.

[0303] In some embodiments, n=3, L C is trihydroxymethylaminometh If it is a tetravalent linker group based on , then the linker is -(L A )3-trihydroxy Methylaminomethane-L B - This allows N-acetylgalactosamine molecules and siRNA molecules to The structure of the siRNA complex formed by binding is shown in formula (304) below.

[0304] [ka] In the formula, the double helix structure represents siRNA.

[0305] Similarly, the complex site between siRNA and the complex group is the 3' or 5' end of the sense strand of siRNA. It may be at the end, or at the 5' end of the antisense strand, or within the internal sequence of the siRNA. That's fine.

[0306] In some embodiments, the 3' end of the sense strand of the siRNA described herein is Linker-(L A )3-trihydroxymethylaminomethane-L B - three N-A It is covalently bound to the cetylgalactosamine (GalNAc) molecule and to the siRNA molecule. The structure of the si molecule with a molar ratio of 1:3 to the GalNAc molecule is shown in the following formula (305). An RNA complex (hereinafter also referred to as (GalNAc)3-siRNA) is obtained.

[0307] [ka] In the formula, the double helix structure represents the siRNA, and the linker represents the sense of the siRNA. It is attached to the 3' end of the chain.

[0308] In some embodiments, when the target group is N-acetylgalactosamine, A suitable linker may have the structure shown in formula (306).

[0309] [ka] During the ceremony, l is an integer between 0 and 3. * This represents the site in the linker that is bonded to the target group by an ether bond. # This represents the site in the linker that binds to siRNA via a phosphate ester bond. .

[0310] In some embodiments, when l=2, the siRNA complex is given by formula (30 It has the structure shown in 7).

[0311] [ka] In the formula, the double helix structure represents the siRNA, and the linker represents the sense of the siRNA. It is attached to the 3' end of the chain.

[0312] The above siRNA complex was synthesized by a method already described in detail in the prior art. This may also be the case. For example, WO2015006740A2 contains a complex of multiple types of siRNAs. The manufacturing method is described in detail. The siR of this disclosure is manufactured by a method well known to those skilled in the art. A NA complex is obtained. For example, in WO2014025805A1, as shown in formula (305) The method for preparing the structure is described, and Rajeev et al. published ChemBioChem 20 Sections 15, 16, and 903-908 describe the method for preparing the structure shown in formula (307).

[0313] In some embodiments, the siRNA complex has a structure shown in formula (308). It holds.

[0314] [ka] During the ceremony, n1 is an integer selected from 1 to 3, and n3 is an integer selected from 0 to 4. , m1, m2, or m3 are independent integers selected from 2 to 10. R 10 , R 11 , R 12 , R 13 , R 14 or R 15 Each of them is independently H. , or C1-C 10 Alkyl alkyl group, C1-C 10 Alkyl halogens and C1-C 10 a Selected from the group consisting of lucoxy groups, R3 is the base of the structure shown in formula A59.

[0315] [ka] In the formula, E1 is OH, SH, or BH2, and Nu is the siRNA of the present disclosure.

[0316] R2 is a linear alkylene group of carbon atoms with a length of 1 to 20, where one or more carbon atoms Atoms are C(O), NH, O, S, CH=N, S(O)2, C2-C 10 Alkenylene group , C2-C 10 Alkynylene group, C6-C 10 Arylene group, C3-C 18 Heterocycline Lennox group and C5-C 10 One or more arbitrary groups selected from the group consisting of heteroarylene groups. It is replaced with, and R2 is C1-C 10 Alkyl alkyl group, C6-C 10 Aryl group, C5-C 10 Heteroaryl group, C1-C 10 Alkyl halogens, -OC1-C 10 alkyl group, -OC1-C 10 Alkylphenyl group, -C1-C 10 alkyl-OH, -OC1-C1 0. Alkyl halogenated group, -SC1-C 10 Alkyl, -SC1-C 10 Alkyl fer Nyl group, -C1-C 10 Alkyl-SH, -SC1-C 10 Alkyl halogens, halo Gen substituent, -OH, -SH, -NH2, -C1-C 10 Alkyl-NH2,-N(C1 -C 10 (Alkyl group)(C1-C 10 Alkyl(alkyl group), -NH(C1-C 10 (Alkyl group) , N(C1-C 10 (Alkyl group)(C1-C 10 Alkylphenyl group), NH(C1-C 10 Alkylphenyl group), cyano group, nitro group, -CO2H, -C(O)O(C1-C 10 Alkyl(C1-C) 10 (Alkyl group)(C1-C10 (Alkyl alkyl group), -CONH(C1-C 10 Alkyl(alkyl group), -CONH2, -NHC(O)(C1-C 10 Alkyl group), -NHC(O)(phenyl group), -N(C1-C 10 Alkyl(C)(O )(C1-C 10 Alkyl(C1-C) 10 Alkyl group)C(O)(phenyl group) ), -C(O)C1-C 10 Alkyl group, -C(O)C1-C 10 Alkylphenyl group, -C(O)C1-C 10 Haloalkyl group, -OC(O)C1-C 10 alkyl groups, -SO 2(C1-C 10 Alkyl group), -SO2 (phenyl group), -SO2 (C1-C 10 Hello (Alkyl ionized), -SO2NH2, -SO2NH(C1-C 10 (alkyl group), -S O2NH (phenyl group), -NHSO2(C1-C 10 Alkyl(alkyl group), -NHSO2(F (enyl group) and -NHSO2(C1-C 10 Any of the group consisting of halogenated alkyl groups It may optionally have one or more substituents. Each L1 is a linear alkylene group of carbon atoms with a length of 1 to 70, where one or more carbon atoms Elementary atoms are C(O), NH, O, S, CH=N, S(O)2, C2-C 10 Alkenirene group, C2-C 10 Alkynylene group, C6-C 10 Arylene group, C3-C 18 Heterocycline Lylene group and C5-C 10 One or more selected from the group consisting of heteroarylene groups It is replaced with C1-C 10 Alkyl alkyl group, C6-C10 Aryl group, C5-C1 0 heteroaryl group, C1-C 10 Alkyl halogens, -OC1-C 10 alkyl group -OC1-C 10 Alkylphenyl group, -C1-C 10 alkyl-OH, -OC1-C 10 Alkyl halogens, -SC1-C 10 Alkyl, -SC1-C 10 Alkylph henyl group, -C1-C 10 Alkyl-SH, -SC1-C 10 Halide alkyl groups, rogene substituent, -OH, -SH, -NH2, -C1-C 10 Alkyl-NH2,-N(C 1-C 10 (Alkyl group)(C1-C 10 Alkyl(alkyl group), -NH(C1-C 10 alkyl group ), N(C1-C 10 (Alkyl group)(C1-C 10 Alkylphenyl group), NH(C1- C 10 Alkylphenyl group), cyano group, nitro group, -CO2H, -C(O)O(C1- C 10 Alkyl(C1-C) 10 (Alkyl group)(C1-C 10 (Alkyl group) -CONH(C1-C 10 Alkyl(alkyl group), -CONH2, -NHC(O)(C1-C1 0 alkyl group), -NHC(O) (phenyl group), -N(C1-C 10 (Alkyl group)C( O)(C1-C 10 Alkyl(C1-C) 10 (alkyl group)C(O)(phenyl Base), -C(O)C1-C 10 Alkyl group, -C(O)C1-C 10 alkylphenyl group , -C(O)C1-C 10 Haloalkyl group, -OC(O)C1-C 10 alkyl group, -S O2(C1-C 10 Alkyl group), -SO2 (phenyl group), -SO2 (C1-C 10 Ha (logenogenic alkyl group), -SO2NH2, -SO2NH(C1-C 10 (Alkyl alkyl group), - SO2NH (phenyl group), -NHSO2 (C1-C 10 Alkyl(alkyl group), -NHSO2( Phenyl group) and -NHSO2(C1-C 10 The group consisting of halogenated alkyl groups It may optionally have one or more substituents.

[0317] In some embodiments, L1 is made up of a base of A1 to A26 or any combination thereof. The groups may be selected from the following, and the structures and definitions of A1 to A26 are as follows.

[0318] [ka] In the formula, j1 is an integer between 1 and 20, and j2 is an integer between 1 and 20. R' is C1-C 10 It is an alkyl group, Ra is selected from the group consisting of the bases of formulas A27 to A45 or any combination thereof.

[0319] [ka] Rb is C1-C 10 It is an alkyl group, JPEG0007887201000023.jpg12124 represents the site where groups are covalently bonded.

[0320] For convenience, L1 is defined as a linear alkylene group, but for example, the above substitution and / or The amine or alkenyl groups resulting from substitution may not be linear groups, or they may have different names. It will be understood by those skilled in the art that there are two lengths of L1. This is the number of atoms in the chain that connects the bond points. For this purpose, the carbon of the straight-chain alkylene A ring obtained by substituting elementary atoms (for example, heterocyclylene or heteroarylene) is 1 Let's consider them as individual atoms.

[0321] M1 represents the target group, and its definition and selectable range are the same as those for the target group described above. How many? In that embodiment, each M1 is an asial glycoprotein on the surface of mammalian hepatocytes. It is one of the ligands that is independently selected from those that have affinity for the receptor.

[0322] M1 has affinity for asialoglycoprotein receptors on the surface of mammalian hepatocytes. If it is a ligand, in some embodiments n1 is an integer from 1 to 3. n3 may also be an integer from 0 to 4, and the M1 target in the siRNA complex Ensure that the number of bases is at least 2. In some embodiments, n1+n 3 ≥ 2, which means that the number of M1 target groups is at least 3, and the M1 target groups and liver surface The siRNA complex more easily binds to the asialoglycoprotein receptor, and furthermore, the siRNA complex Promoting the uptake of substances into cells through endocytosis (intracellular uptake). This can be done. As can be seen from the experiment, if the number of M1 target groups is 3 or more, the M1 target groups Since the improvement in the ease of binding to asialoglycoprotein receptors on the liver surface is not clear, Considering various factors such as ease of synthesis, structure / process cost, and delivery efficiency, In some embodiments, n1 is an integer between 1 and 2, n3 is an integer between 0 and 1, and Therefore, n1 + n3 = 2 to 3.

[0323] In some embodiments, m1, m2, or m3 are independently selected from integers between 2 and 10. If selected, the spatial position between multiple M1 target groups is determined by the relationship between the M1 target group and the asialoglycoprotein on the liver surface. The siRNA complex provided by this disclosure can be adapted to bind to nucleotide receptors. To simplify the body, make it easier to synthesize, and / or reduce the cost, several In this embodiment, m1, m2, and m3 are each an integer between 2 and 5, and In some embodiments, m1 = m2 = m3.

[0324] R 10 , R 11 , R 12 , R 13 , R 14 or R 15 However, H, C1-C 10 alkyl group , C1-C 10 Alkyl halogens and C1-C 10 Each alkoxy group is independent If one is selected, neither alters the properties of the siRNA complex of this disclosure. Those skilled in the art will understand that the objectives of this disclosure can be achieved in any way. In this embodiment, R 10 , R 11 , R 12 , R 13 , R 14 or R 15 These are, Independently selected from H, a methyl group, or an ethyl group. In some embodiments, R1 0, R 11 , R 12 , R 13 , R 14 and R 15All of these are H.

[0325] R3 is the base of the structure shown in formula A59, where E1 is OH, SH, or BH2. In some embodiments, considering the availability of the raw materials for preparation, E1 is OH or It is SH.

[0326] R2 is selected to enable bonding between the N atom on the nitrogen-containing skeleton and A59. In the context of the detailed explanation, "nitrogen-containing skeleton" means R 10 , R 11 , R 12 , R 13 , R 14 and R 15 This refers to a chain-like structure in which carbon atoms and N atoms are bonded to each other. Therefore, R2 can bond the A59 group to the N atom on the nitrogen-containing skeleton in an appropriate manner. Any linker group may be used. In some embodiments, by a solid-phase synthesis process When preparing the siRNA complex shown in formula (308), the R2 group contains nitrogen-containing bone It contains both a site that bonds to the higher-ranking N atom and a site that bonds to the P atom in R3. It is necessary to do so. In some embodiments, the N atoms on the nitrogen-containing skeleton in R2 The bonded portion forms an amide bond with the N atom and the bonded portion is to the P atom on R3. The position forms a phosphate ester bond with the P atom, and in some embodiments, R2 is B 5, B6, B5', or B6' may also be used.

[0327] [ka] During the ceremony, JPEG0007887201000025.jpg12136 represents the site where groups are covalently bonded.

[0328] The range of the value of q2 may be an integer from 1 to 10, and in some embodiments, q 2 is an integer between 1 and 5.

[0329] L1 is bonded to the M1 target group and N on the nitrogen-containing skeleton, forming the siRN shown in formula (308). It plays a role in providing liver targeting function to the A complex. In some embodiments, L 1 is a combination of one or more bonds selected from the bases of formulas A1 to A26. In this embodiment, L1 is A1, A4, A5, A6, A8, A10, A11 and A13 It is a combination of one or more combinations selected from. In some embodiments, L1 is , in a combination of at least two joins selected from A1, A4, A8, A10 and A11 Yes. In some embodiments, L1 is selected from A1, A8, A10. Both are combinations of two bonds.

[0330] In some embodiments, the length of L1 is 3 to 25 atoms, 3 to 20 atoms, The number of atoms may be 4 to 15 or 5 to 12. In some embodiments, The length of L1 is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 pieces. 13 pieces, 14 pieces, 15 pieces, 16 pieces, 17 pieces, 18 pieces, 19 pieces, 20 pieces, 21 pieces, 22 pieces, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60 It is an atom.

[0331] In some embodiments, j1 is an integer from 2 to 10, and in some embodiments... In some embodiments, j1 is an integer between 3 and 5. In some embodiments, j2 is an integer between 2 and 10. In some embodiments, j2 is an integer between 3 and 5. R' is C1-C4. It is a methyl group, and in some embodiments, R' is a methyl group, an ethyl group, and an isopyl group. It is one of the ropyl groups. Ra is one of A27, A28, A29, A30 and A31. In some embodiments, Ra is A27 or A28. Rb is C1-C It is a 5-alkyl group, and in some embodiments, Rb is a methyl group, an ethyl group, or an iso It is one of the propyl group and the butyl group. In some embodiments, formulas A1 to A26 By selecting j1, j2, R', Ra, and Rb respectively, the M1 target group and nitrogen This achieves bonding with the N atom on the contained skeleton, and the spatial position between the M1 target groups is determined between the M1 target group and the liver surface. Further improves the binding to the asialoclycoprotein receptor.

[0332] In some embodiments, the siRNA complex is of formula (403), (404), (405), (406), (407), (408), (409), (410), (411 ), (412), (413), (414), (415), (416), (417), (4 It has the structure shown in 18), (419), (420), (421), or (422).

[0333] [ka] JPEG0007887201000027.jpg205132JPEG0007887201000028.jpg206131JPEG0007887201000029.jpg194120 JPEG0007887201000030.jpg233118JPEG0007887201000031.jpg214115JPEG0007887201000032.jpg162121

[0334] In some embodiments, the P atom in formula A59 is a different atom in the siRNA sequence. It may be bonded to any possible position; for example, the P atom in formula A59 is of the siRNA. It may be bound to one nucleotide of either the sense strand or the antisense strand, and any number In that embodiment, the P atom in formula A59 is any one of the sense strands of the siRNA. In some embodiments, the P atom in formula A59 It is bound to the end of the sense or antisense strand of the siRNA, and in some embodiments In formula A59, the P atom is bound to the end of the sense strand of the siRNA. The part refers to the four nucleos in the sense chain or antisense chain from one end to the front. It refers to siRNA. In some embodiments, the P atom in formula A59 is the siRNA. Bonded to the end of an antisense chain or antisense chain, in some embodiments, formula A59 The P atom is attached to the 3' end of the sense strand of the siRNA. When bound to the above position, the siRNA complex represented by formula (308) enters the cell. After being inserted, when it is unwound, it releases the antisense strand of a single siRNA, and XO It is possible to block the process by which mRNA translates into protein, thereby suppressing XO gene expression. .

[0335] In some embodiments, the P atom in formula A59 is a nucleus in siRNA. Any possible position on the ocide, for example, the 5' position of the nucleotide, the 2' position of the nucleotide, It may be attached to the 3' position of the nucleotide or to a base of the nucleotide. Several embodiments In this, the P atom in formula A59 forms a phosphate diester bond, thereby It may be bound to the 2', 3', or 5' position of the nucleotide in siRNA. In several embodiments, the P atom in formula A59 is the 3' end of the sense strand of the siRNA. The 3' hydroxyl group of the nucleotide is bonded to the oxygen atom from which it was dehydrogenated (at this time, A59 The P atom in this can also be considered as the P atom of the phosphate group contained in siRNA), or In formula A59, the P atom is one nucleotide in the sense strand of siRNA. '-By substituting a hydrogen in the hydroxyl group, it is bound to the nucleotide, or, formula A The P atom at 59 is the 5' hydroxyl group of the 5' terminal nucleotide of the sense strand of siRNA. It is bonded to a nucleotide by substituting a hydrogen atom in the group.

[0336] The inventors of this disclosure claim that the siRNA complex of this disclosure exhibits significantly improved stability in plasma and is off The target effect is reduced, and high XO mRNA silencing activity is also present. We unexpectedly found that the following was observed. In some embodiments, the siRNA of this disclosure is It may also be one of the siRNAs shown in Tables 1a to 1l. The siRNA complex exhibits higher XO mRNA silencing activity.

[0337] [Table 1a]

[0338] [Table 1b]

[0339] [Table 1c]

[0340] [Table 1d]

[0341] [Table 1e]

[0342] [Table 1f]

[0343] [Table 1g]

[0344] [Table 1h]

[0345] [Table 1i]

[0346] [Table 1j]

[0347] [Table 1k]

[0348] [Table 1l] However, uppercase C, G, U, and A represent the base sequence of a nucleotide, and lowercase m represents the same This indicates that the nucleotide adjacent to the left of the letter 'm' is a methoxy-modified nucleotide. Furthermore, the lowercase letter f is a fluoromodified nucleus, where one nucleotide adjacent to the left of the letter f is fluoromodified. The lowercase 's' indicates that there is a thiol between the two nucleotides to the left and right of the letter. This indicates that the molecules are bonded by an ester group, and P1 is the molecule adjacent to the right of P1. One nucleotide is a 5'-phosphate nucleotide or a 5'-phosphate analog modified nucleotide. It indicates that it is a do. In some embodiments, P1 is a specific modifier of VP, Ps or represents P, and the combination character VP is one adjacent to the right of the combination character VP. The nucleotide is a vinyl phosphate ester (5'-(E)-vinylphosphonat e, E-VP) indicates a modified nucleotide, and the combination character Ps is the same combination character The nucleotide adjacent to the right of Ps is a thiophosphate-modified nucleotide. This indicates that the capital letter P is a nucleotide adjacent to the right of the letter P, specifically a 5'-phosphorus nucleotide. This indicates that it is an acid nucleotide.

[0349] In the siRNA or siRNA complex described in this disclosure, each adjacent nucleoti The bonds between the phosphates are connected by phosphate diester bonds or thiophosphate diester bonds, In a sterling bond or thiophosphate diester bond, the non-crosslinked oxygen or sulfur atom is negatively charged. It carries a charge and has a hydroxyl group or sulfhydryl group. They may exist as such, and the hydrogen ions in the hydroxyl group or sulfhydryl group may be partially or The entire molecule may be substituted with a cation. The cation is any cation, for example, a metal. Cation, ammonium ion NH4 + Even if it is one of the organic ammonium cations i. Considering the improvement of solubility, in one embodiment, the cation is an alkali metal i On, ammonium cations formed by tertiary amines and quaternary ammonium catios It is one or more types selected from the following. Alkali metal ions are K+ and / or Na + It may also be the case that the cation formed by the tertiary amine is formed by triethylamine. Formed by ammonium ions and / or N,N-diisopropylethylamine The ammonium ion may be formed. Therefore, the siRN described in this disclosure A or the siRNA complex may exist as a salt in at least a portion of it. In one embodiment... Furthermore, non-crosslinked oxygen atoms in the phosphate diester bond or thiophosphate diester bond or At least a portion of the sulfur atom is bonded to a sodium ion, as described in this disclosure. NA or siRNA complexes exist as sodium salts or partial sodium salts.

[0350] As will be obvious to those skilled in the art, nucleoside monomas having the corresponding modifications By using -, modified nucleotide groups can be introduced into the siRNA described in this disclosure. This can be done. A method for preparing nucleoside monomers having the corresponding modifications, and modified nucleos Methods for introducing rheotide groups into siRNA are also well known to those skilled in the art. Rheoside monomers may be purchased commercially or prepared by known methods.

[0351] <Preparation of the siRNA complex shown in formula (308)> Even if the siRNA complex shown in formula (308) is prepared by any reasonable synthetic route, good.

[0352] In some embodiments, the siRNA complex represented by formula (308) is as follows: It can be prepared by the method. This method is performed under the conditions of phosphoramidite solid-phase synthesis. Depending on the nucleotide types and order of the sense and antisense strands of each siRNA, 3' Nucleoside monomers are sequentially bonded from the 5' end, and the bonding of each nucleoside monomer... This involves four reactions: deprotection, coupling, capping, oxidation, or sulfidation, and siRNA The process includes isolating the sense strand and antisense strand of the siRNA and annealing them, wherein the siRNA is , the siRNA of the present disclosure described above.

[0353] Furthermore, this method, under coupling reaction conditions and in the presence of a coupling reagent, is performed using formula (32 1) The compound shown in 1) is bound to a nucleoside monomer or a nucleotide on a solid support. The sequence is brought into contact with the compound shown in formula (321) and coupled to form a nucleotide. This further includes binding to the sequence. Below, the compound shown in formula (321) is a complex molecule. It is also called [another name].

[0354] [ka] During the ceremony, R4 can bind to the siRNA represented by Nu in the compound shown in formula (308). It is a group. In some embodiments, R4 is a si represented by covalent bonding to Nu. It is a group that can bind to RNA. In some embodiments, R4 reacts to form a phosphate group. It is a group that can be compounded with any functional group of siRNA represented by Nu via a diester bond. In each S1, all active hydroxyl groups in M1 are independently replaced with YCOO- groups. The group is a methyl group, trifluoromethyl group, difluoromethyl group, fluoromethyl group, and fluoromethyl group. Tyl group, trichloromethyl group, dichloromethyl group, chloromethyl group, ethyl group, n-pro Independently of pyr group, isopropyl group, phenyl group, halophenyl group and alkylphenyl group One of the selected elements is a methyl group, and in some embodiments, Y is a methyl group.

[0355] n1, n3, m1, m2, m3, R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , The definitions and selectable ranges for L1 and M1 are as described above.

[0356] R4 forms a bond with the N atom on the nitrogen-containing skeleton, resulting in the siRN shown in formula (308). Selected to provide a suitable reaction site for the synthesis of complex A. And R4 is either an R2 linker group or a protected R2 linker group, and reacts It contains functional groups that can form the structure shown in siRNA and A59.

[0357] In some embodiments, R4 is represented by Nu, which is an siRNA or nucleoside. A first functional group that can form a phosphite ester with a group on the nomer, and a hydroxyl group Alternatively, a second functional group that can react with an amino group to form a covalent bond or the covalent bond It includes a solid support bonded by bonding. In some embodiments, the first functional group These are phosphoramidites, hydroxyl groups, or protected hydroxyl groups. In embodiments, the second functional group is a phosphoramidite, a carboxyl group, or a carbo It is a phosphate salt. In some embodiments, the second functional group is separated by a covalent bond. A solid support bonded to the rest of the child, wherein the covalent bond is a hydroxyl group or an amino group It is formed by phosphate ester. In some embodiments, the solid phase support is phosphate ester They are bonded via carboxylic acid ester bonds or amide bonds. Several implementations In terms of form, the solid-phase carrier is a resin.

[0358] In some embodiments, the first functional group is a hydroxyl group, -OR k or formula ( The group is represented by C3), and the second functional group is represented by formula (C1), (C2), (C3), ( Includes the structure shown in C1' or (C3').

[0359] [ka] In the formula, q1 is an integer from 1 to 4, X is O or NH, and M + R is a cation, k is a hydroxy protecting group, and SPS represents a solid support. JPEG0007887201000047.jpg11126 represents the site where the group is bonded to the covalent bond.

[0360] In some embodiments, the first functional group is as shown in formula (C3), It contains a foramidite group, and the phosphoramidite group is at any position on the nucleotide Coupling with a hydroxyl group, for example, a hydroxyl group at the 2' position or a hydroxyl group at the 3' position. The reaction forms a phosphite ester, which is then oxidized or sulfurized to form a phosphate diphosphate represented by formula A59. By forming an ester bond or thiophosphate ester bond, the complex molecule can be combined with siRNA. This is possible. In this case, even if the second functional group is not present, the compound of formula (321) is It can be compounded with nucleotides, and is used to obtain the siRNA complex shown in formula (308). It will not have any effect. In this case, siRN can be synthesized by methods such as phosphoramidite solid-phase synthesis. After obtaining the sense strand or antisense strand of A, the compound of formula (321) and the nucleotide sequence In the subsequent oxidation or sulfurization process, the hydroxyl group on the terminal nucleotide is reacted, and in the subsequent oxidation or sulfurization process... A bond is formed by a phosphate diester bond or a thiophosphate ester bond, and the formula (321) The compound is combined with the siRNA.

[0361] In some embodiments, the first functional group comprises a protected hydroxyl group. In some embodiments, the second functional group includes a group that can react with a solid support, and The reaction provides a composite molecule containing a solid support. In some embodiments, the The functional group of 2 is a carboxyl group, as shown in formula (C1), (C2), or (C3). The second functional group is a carboxyl group or a carboxyl group. When containing hydroxyl salts, the compound of formula (321) and the solid support, for example, hydroxyl salts in resins. The group or amino group is subjected to an esterification or amidation reaction, and a carboxylic acid ester bond is formed The bonded, solid-phase support forms a composite molecule. The second functional group is phosphoramide When it contains a functional group, the compound of formula (321) and a general-purpose solid support, for example, a resin The hydroxy group is subjected to a coupling reaction, oxidized, and bonded by a phosphate diester bond. Then, a composite molecule containing a solid support is formed. Subsequently, the product to which the solid support is bound is used as a starting point. Then, following the phosphoramidite solid-phase synthesis method, nucleoside monomers are sequentially bonded, forming a complex Obtain the sense or antisense strand of siRNA to which the group is attached. During the phase synthesis process, the first functional group is deprotected, and then the coupling reaction conditions are met. Below, it is coupled with the phosphoramidite group in the nucleoside monomer.

[0362] In some embodiments, the first functional group is a hydroxyl group or a protected hydroxyl group The roxy group is included, and the second functional group is as shown in formula (C1') or (C3'), Solid support bonded by carboxylic acid ester bonds or solid phase bonded by amide bonds It includes a carrier, or a solid-phase carrier bonded by phosphate ester bonds. In this case, as Using the compound of formula (321) instead of a solid support, according to the phosphoramidite solid-phase synthesis method The sense strand or a complex group of siRNA is formed by sequentially linking nucleoside monomers. Obtain an inthense chain.

[0363] In some embodiments, the carboxylate salt is -COO - M + It may also be expressed as this, Here, M + These are cations, such as metal cations and ammonium cations (NH4). + , It is one selected from organic ammonium cations. In one embodiment, the gold The group ion is one selected from alkali metal ions, for example, K + or Na + in Yes. In some embodiments, taking into consideration the need to improve solubility and facilitate the reaction, In this case, the organic ammonium ion is formed by tertiary amines, ammonium catio Ammonia formed by a quaternary ammonium cation, such as triethylamine. Ammonium ions formed by ammonium ions or N,N-diisopropylethylamine In some embodiments, the carboxylate salt is triethylaminecarboxylic acid. It is a salt or an N,N-diisopropylethylamine carboxylate salt.

[0364] In some embodiments, R4 is derived from formulas (B9), (B10), (B9'), and (B1 Includes the structure shown in (0'), (B11), (B12), (B11'), or (B12'). .

[0365] [ka] In the formula, q1 is an integer between 1 and 4, q2 is an integer between 1 and 10, and X is either O or NH. Ri, M + R is a cation, k is a hydroxy protecting group, and SPS represents a solid support. JPEG0007887201000049.jpg10140 represents the site where the group is bonded to the covalent bond. In some embodiments, q1 is 1 or 2. In some embodiments, q2 is an integer from 1 to 5. In the application form, R4 includes the structure shown in formula (B9) or (B10). In the embodiment, R4 includes the structure shown in formula (B11) or (B12).

[0366] In some embodiments, R k This is Tr (trityl group), MMTr (4-methoxy Trityl group), DMTr (4,4'-bismethoxytrityl group), TMTr (4,4', It is one or more 4''-trimethoxytrityl groups. In some embodiments, R k This is DMTr, i.e., 4,4'-bismethoxytrityl(4,4'-dimetho (xytrityl) is also acceptable.

[0367] The definition of L1 is as described above.

[0368] In some embodiments, L1 bonds the M1 target group to the N atom on the nitrogen-containing skeleton. It is used to provide liver targeting function to the siRNA complex shown in formula (308). In some embodiments, L1 includes one of A1 to A26 or a combination thereof. nothing.

[0369] As can be easily understood by those skilled in the art from the above description, phosphorus known in the art Compared to the midite solid-phase synthesis method, the above first functional group and an arbitrary second functional group allow for multiple synthesis. The compound molecule can be placed at any possible position in the nucleotide sequence, for example, at the ends of the nucleotide sequence. Obtain an siRNA complex represented by formula (308) bound to the end of a rheotide sequence. This is possible. Accordingly, unless otherwise specified, the following refers to siRNA complexes and / or compound complexes. In descriptions concerning the preparation of molecules, the terms "deprotection," "coupling," and "capping" are used. When referring to reactions such as "oxidation" and "sulfidation," the phosphoramidite nucleic acids known in this field... The reaction conditions and reagents related to solid-phase synthesis methods should also be understood to apply to these reactions. The exemplary reaction conditions and reagents are described in detail below.

[0370] In some embodiments, each S1 is independently M1. In each S1, at least one active hydroxyl group is present in M1. It is a group protected by a protective group. In some embodiments, each S1 is independently M1 All of the active hydroxyl groups present are protected by hydroxyl protecting groups. In embodiments, any hydroxy protecting group known to those skilled in the art is used to activate the hydroxyl group at M1. It can be used to protect the roxy group. In some embodiments, it is protected. The hydroxyl group may also be represented by the formula YCOO-, where each Y is independently C1-C 10 Al Kill group and C6-C 10 Selected from the group consisting of aryl groups, the C1-C 10 Alkyl Base and C6-C 10 The aryl group is optionally substituted with one or more substituents, and the substituents are Selected from the group consisting of halogens and C1-C6 alkyl groups. In some embodiments In this structure, each Y independently comprises a methyl group, a trifluoromethyl group, a difluoromethyl group, and a mo Nofluoromethyl group, trichloromethyl group, dichloromethyl group, chloromethyl group, ethyl Groups, n-propyl groups, isopropyl groups, phenyl groups, halophenyl groups and C1-C6 groups Selected from the group consisting of chylphenyl groups.

[0371] In some embodiments, each S1 independently consists of formulas A46 to A54. Selected from the group.

[0372] [ka]

[0373] In some embodiments, S1 is formula A49 or A50.

[0374] In some embodiments, each Y is a methyl group, a trifluoromethyl group, a difluoromethyl group. Methyl group, fluoromethyl group, trichloromethyl group, dichloromethyl group, chloromethyl group ethyl group, n-propyl group, isopropyl group, phenyl group, halophenyl group and alkyl One independently selected from the phenyl group, and in some embodiments, Y is me It is a chill group.

[0375] As mentioned above, the method for preparing the siRNA complex shown in formula (308) is as follows: Synthesize the other chain (for example, the sense chain of the siRNA to which the complex molecule was attached in the above step). When synthesizing, further synthesize the siRNA antisense strand according to the solid-phase synthesis method. (Included in, and vice versa), isolate the sense strand and antisense strand, and anneal them. The process further includes the following steps: Specifically, in the isolation step, the nucleotide sequence and / or The solid support bonded to the composite molecule is cleaved, and the necessary protecting groups are removed (at this point In combination, each S1 group in the compound of formula (321) is converted to the corresponding M1 target group. The sense strand (or antisense strand) of the siRNA to which the synthesizer is bound and the corresponding antisense strand A sense strand (or antisense strand) is obtained, and the sense strand and antisense strand are annealed to form a double-stranded RN. Structure A is formed, and an siRNA complex shown in formula (308) is obtained.

[0376] In some embodiments, the method for preparing the siRNA complex shown in formula (308) Under coupling reaction conditions and in the presence of a coupling reagent, the compound represented by formula (321) The substance is brought into contact with the first nucleoside monomer at the 3' end of the sense chain or antisense chain. The compound shown in formula (321) is attached to the first nucleotide in the sequence, and a phosphorus is formed. Under the conditions of ruamidite solid-phase synthesis, seed nucleotides of the desired sense or antisense strand are obtained. Depending on the class and order, nucleoside monomers are bonded sequentially from 3' to 5', forming siRN A step to synthesize a sense chain or antisense chain of A, wherein the compound of formula (321) is R 4. A first functional group containing a protected hydroxyl group, and formula (C1') or (C3') A compound containing a second functional group having the structure shown, the first nucleoside mono Before bonding with Mer, the compound of formula (321) is deprotected, and each nucleoside monomer is bonded. This involves four reactions: deprotection, coupling, capping, oxidation, or sulfidation, and the complex group is bonded. A step to obtain a sense or antisense strand of a combined nucleic acid; solid-phase synthesis of phosphoramidites Depending on the type and order of nucleotides in the antisense or sense strand, the 3' to 5' Nucleoside monomers are sequentially bonded toward the antisense or sense strand of the nucleic acid. The bond between each nucleoside monomer undergoes deprotection, coupling, capping, oxidation, or The process involves four sulfidation reactions, removal of protecting groups, cleavage from the solid support, isolation and purification to obtain the sense chain. The process also includes obtaining an antisense chain and performing an annealing step.

[0377] In some embodiments, the method for preparing the siRNA complex shown in formula (308) , the type and order of nucleotides in the sense strand or antisense strand of the double-stranded siRNA. In order, nucleoside monomers are bonded sequentially from 3' to 5', and the sense chain and A The nucleosine chain is synthesized, and the bonds of each nucleoside monomer undergo deprotection, coupling, and capping. The process involves four reactions: ping, oxidation, or sulfurization, and includes a sense chain bonded to a solid support, and the solid support Steps to obtain an antisense chain bound to a body; coupling reaction conditions and coupling reagents In the presence of the compound shown in formula (321), the sense chain or solid phase bonded to the solid support The antisense chain bound to the support is brought into contact with the first phosphoramidite group at R4. A compound of formula (321) containing a functional group is attached to a sense chain or antisense chain for protection. The groups were removed, the solid support was cleaved, and each was isolated and purified to obtain siRNA with the complex group attached. The process includes obtaining a sense chain or antisense chain and performing an annealing step.

[0378] In some embodiments, the P atom in formula A59 is a sense atom in siRNA. The method for preparing the siRNA complex shown in formula (308), which is bound to the 3' end of the chain, is as follows: (1) Compound of formula (321) (The compound of formula (321) has a protected hydroxygen at R4. C base OR k A first functional group containing, and having a structure represented by formula (C1') or (C3') (A compound containing a second functional group) with a hydroxy protecting group R k Remove, cut Under pulling reaction conditions and in the presence of a coupling reagent, the deprotected product is converted to a nucleoside. By contacting the monomer, a nucleoside monomer is obtained that is bonded to a solid support via a composite molecule. thing, (2) Starting with a nucleoside monomer that is bonded to a solid support by the composite molecule, 3 The sense strand of siRNA was synthesized in the '-5' direction using a phosphoramidite solid-phase synthesis method. to do, (3) Synthesize the antisense chain of siRNA by a phosphoramidite solid-phase synthesis method. to (4) The sense strand and antisense strand of the siRNA are isolated and annealed, and formula (308) This includes obtaining the siRNA complex shown in [reference].

[0379] In step (1), the protecting group R in the compound of formula (321) above k How to remove This includes contacting the compound of formula (321) with a deprotection reagent under deprotection conditions. The temperature is 0-50°C, and in some embodiments, 15-35°C, and the reaction time is 30 ~300 seconds, 50~150 seconds in some embodiments, and the deprotection reagent is trifluor. One or more selected from chloroacetic acid, trichloroacetic acid, dichloroacetic acid, and chloroacetic acid. It may also be dichloroacetic acid in some embodiments. Deprotection agent and formula (321) The molar ratio with the compound is 10:1 to 1000:1, and in some embodiments it is 50:1 to The ratio is 500:1.

[0380] The coupling reaction conditions and coupling reagents are suitable for the above coupling reaction. Any conditions and reagents may be used. In some embodiments, solid-phase synthesis is employed. The same conditions and reagents as those used in the coupling reaction may be employed.

[0381] In some embodiments, the conditions for the coupling reaction are such that the reaction temperature is 0 to 50°C. In some embodiments, the temperature is 15-35°C. Compounds of formula (321) and nucleosides The molar ratio with nomer is 1:1 to 1:50, and in some embodiments it is 1:2 to 1:5. Yes, the molar ratio of the compound in formula (321) to the coupling reagent is 1:1 to 1:50. In some embodiments, the ratio is 1:3 to 1:10, and the reaction time is 200 to 3000. The time is 500 to 1500 seconds, in some embodiments. The coupling reagent is 1H-T Trazol, 5-ethylthio-1H-tetrazol, 5-benzylthio-1H-tetrazol One or more selected from, and in some embodiments, 5-ethylthio-1H-the It is trazole. The coupling reaction may be carried out with an organic solvent, and the organic solvent This is one or more selected from anhydrous acetonitrile, anhydrous DMF, and anhydrous dichloromethane. The species is anhydrous acetonitrile in some embodiments. In contrast, the amount of the organic solvent is 3 to 50 L / mol, and in some embodiments, 5 to 20 L. It is / mol.

[0382] In step (2), the phosphoramidite nucleic acid prepared in the above step is synthesized by a solid-phase synthesis method. Starting with a nucleoside monomer bonded to a solid support by a composite molecule, 3'-5' In this direction, the sense strand SS of the second siRNA complex is synthesized. In this case, the complex group is obtained It is attached to the 3' end of the sense strand.

[0383] Other conditions for the solid-phase synthesis in steps (2) and (3) include the removal of nucleoside monomers. Protection conditions, type and dosage of deprotection reagent, coupling reaction conditions, type of coupling reagent and Dosage, conditions for the capping reaction, type and dosage of the capping reagent, conditions for the oxidation reaction, oxidation reagent This includes the type and dosage of the drug, the sulfurization reaction conditions, and the type and dosage of the sulfurization reagent, which are commonly used in this field. Various reagents, dosages, and conditions will be adopted.

[0384] For example, in some embodiments, in steps (2) and (3), the solid-phase synthesis Then, the following conditions may be used.

[0385] The deprotection conditions for nucleoside monomers are a temperature of 0-50°C, and in some embodiments, 1 The temperature is 5-35°C, and the reaction time is 30-300 seconds, or 50-150 seconds in some embodiments. The deprotection reagents are trifluoroacetic acid, trichloroacetic acid, dichloroacetic acid, and chloroacetic acid. It may be one or more selected from, and in some embodiments, dichloroacetic acid Yes. The molar ratio of the deprotection reagent to the 4,4'-dimethoxytrityl protecting group in the solid support is The ratio may be 2:1 to 100:1, and in some embodiments it is 3:1 to 50:1.

[0386] The coupling reaction conditions are a temperature of 0-50°C, and in some embodiments, 15-35°C. Yes, the molar ratio of the nucleic acid sequence bound to the solid support to the nucleoside monomer is 1:1 to 1: It may be 50, and in some embodiments it is 1:5 to 1:15, and is bonded to a solid support. The molar ratio of the nucleic acid sequence to the coupling reagent is 1:1 to 1:100, according to several embodiments. Then it is 1:50 to 1:80, and the reaction time and selection of coupling reagent are the same as above. ru.

[0387] The capping reaction conditions are a temperature of 0-50°C, and in some embodiments, 15-35°C. Yes, the reaction time is 5 to 500 seconds, and in some embodiments it is 10 to 100 seconds, and the catch The selection of the capping reagent is the same as described above. The total amount of capping reagent and the amount bound to the solid support. The molar ratio of the nucleic acid sequence is 1:100 to 100:1, and in some embodiments, 1:10 to 1 The ratio is 0:1. Equimolar amounts of anhydride acetate and N-methylimidazole are used as the capping reagent. When using acetic anhydride, N-methylimidazole, and nucleic acid sequence bound to the solid support The molar ratio may be 1:1:10 to 10:10:1, and in some embodiments, 1: The ratio is 1:2 to 2:2:1.

[0388] The oxidation reaction conditions are such that the temperature is 0-50°C, and in some embodiments, 15-35°C. The reaction time is 1 to 100 seconds, and in some embodiments, 5 to 50 seconds, and the oxidizing reagent is several In one embodiment, it is iodine (provided as iodine water in some embodiments). The molar ratio of the chemical reagent to the nucleic acid sequence bound to the solid support in the coupling step is 1: The ratio may be 1 to 100:1, and in some embodiments it is 5 to 50:1. In one embodiment, the oxidation reaction is carried out in a ratio of tetrahydrofuran:water:pyridine = 3:1:1 The reaction is carried out using a mixed solvent of approximately 1:1:3. The sulfurization reaction conditions are a temperature of 0-50°C, with several practical requirements. In the application method, the temperature is 15-35°C, the reaction time is 50-2000 seconds, and in some embodiments, The reaction time is 100 to 1000 seconds, and the sulfiding reagent is xanthan hydride in some embodiments. Yes. The molar ratio of the sulfidation reagent to the nucleic acid sequence bonded to the solid support in the coupling process. The ratio is 10:1 to 1000:1, and in some embodiments it is 10:1 to 500:1. In some embodiments, the sulfurization reaction is carried out with acetonitrile:pyridine = 1:3~3: This is done with a mixed solvent of 1.

[0389] After all nucleoside monomers have been bonded, and before annealing, the method involves siRN The method further includes isolating the sense and antisense strands of A. The isolation method is publicly known to those skilled in the art. It is a known process, and generally, it involves cleaving a synthesized nucleotide sequence from a solid support, and then, on the base, phosphorus... This includes removing protecting groups from acid groups and ligands, purifying the material, and desalting it.

[0390] The synthesized nucleotide sequence is cleaved from the solid support, and then cleaved at the base, phosphate group, and ligand. The protecting group above is removed by the usual cleavage and deprotection methods used in siRNA synthesis. For example, the nucleotide sequence to which the obtained solid support is bound can be measured in concentrated ammonia water. In contact with the hydroxyl group, the protecting group YCOO- of the A46-A54 group is removed during the deprotection process. The S1 group is converted to the corresponding M1 group, and the siRNA compound shown in formula (308) is obtained. A compound is formed. Here, the concentrated ammonia water is 25-30% by weight ammonia water. However, the dosage of concentrated ammonia water is 0.2 ml / of the target siRNA sequence. The concentration may be μmol to 0.8 ml / μmol.

[0391] If the synthesized nucleotide sequence has at least one 2'-TBDMS protection, then The notation method involves recording the nucleotide sequence from which the solid support has been removed using triethylamine hydrofluoric acid. This further includes removing the 2'-TBDMS protection by bringing it into contact with the In this case, the corresponding nucleotide in the obtained target siRNA sequence contains two free nucleotides. It has a hydroxyl group. The dose of pure triethylamine hydrofluoric acid is intended and For the siRNA sequence, a concentration of 0.4 ml / μmol to 1.0 ml / μmol is acceptable. i. Thus, an siRNA complex represented by formula (308) can be obtained.

[0392] Methods of purification and desalting are well known to those skilled in the art. For example, preparative ion chromatography Nucleic acid purification is performed by gradient elution of NaBr or NaCl using a Graphy purification column. After completing the process and collecting and combining the products, desalting is performed using a reverse-phase chromatography purification column. It is possible.

[0393] In the siRNA complex shown in formula (308) obtained in this way, nucleotides Non-crosslinked oxygen atoms or sulfur in the phosphate diester bond or thiophosphate diester bond between them. The yellow atoms are basically bonded to sodium ions, as shown in formula (308) siR The NA complex basically exists as a sodium salt. By well known ion exchange methods... or, the sodium ion is replaced with a hydrogen ion and / or other cation, and other forms of formula An siRNA complex shown in (308) can be obtained. The cation is as described above. That is correct.

[0394] During the synthesis process, the purity and molecular weight of the nucleic acid sequence are constantly detected to better control the synthesis quality. Such detection methods are known to those skilled in the art. For example, ion exchange chromatography Nucleic acid purity is detected by matrixing, and liquid chromatography-tandem mass spectrometry (L) is performed. Molecular weight can be measured by C-MS.

[0395] The annealing method is also well known to those skilled in the art. For example, a easily synthesized annealing Mix the S chain and antisense chain (AS chain) in equimolar ratios with sterile water for injection and use 70-95 By heating to °C and then cooling to room temperature, a double-chain structure can be formed through hydrogen bonding. Thus, an siRNA complex represented by formula (308) can be obtained.

[0396] After obtaining the siRNA complex, in some embodiments, for example, liquid chromatography Formula (30) was synthesized by molecular weight detection using methods such as graphi-tandem mass spectrometry. The characteristics of the siRNA complex shown in 8) were clarified, and the synthesized siRNA complex was The siRNA complex shown in the target design formula (308), as well as the synthesized siR Confirm that the NA sequence is one of the desired siRNA sequences, for example, one of the sequences shown in Table 1. It is also possible.

[0397] The compound shown in formula (321) reacts in organic solvents under esterification conditions and in bases. And in the presence of an esterification catalyst, the compound shown in formula (313) is brought into contact with a cyclic acid anhydride. A preparation method comprising ion exchange and isolation to obtain a compound shown in formula (321). It can be obtained by doing so.

[0398] [ka] In the formula, n1, n3, m1, m2, m3, R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , L 1、 The definitions and selectable ranges for each S1 are as described above. R6 is the group that provides R4 in formula (321). In some embodiments, R6 has the structure shown in formula (A61).

[0399] [ka] In the formula, R i This enables bonding with N atoms on the nitrogen-containing skeleton, R k It combines with O to form one Any group to which the free hydroxyl group is attached, R k This is a hydroxy protecting group. In this case, R4 includes a first functional group and a second functional group as hydroxy protecting groups, and the above A compound of formula (321) is obtained in which the second functional group contains the structure shown in formula (C1) or (C2). It is possible.

[0400] The esterification reaction conditions are a reaction temperature of 0 to 100°C and a reaction time of 8 to 48 hours. In some embodiments, the esterification reaction conditions are such that the reaction temperature is 10 to 40°C. The reaction time is 20 to 30 hours.

[0401] In some embodiments, the organic solvent is an epoxy solvent, an ether solvent, etc. Alkyl chloroformides solvents, dimethyl sulfoxide, N,N-dimethylformamide and N , comprising one or more types of N-diisopropylethylamine. In some embodiments, The epoxy solvent is dioxane and / or tetrahydrofuran, and the The tert solvent is ethyl ether and / or methyl tert-butyl ether, and the above Alkyl halogenated solvents include dichloromethane, trichloromethane, and 1,2-dichloro It is one or more types of ethane. In some embodiments, the organic solvent is dichloro It is lomethane. For the compound shown in formula (313), the amount of the organic solvent is 3 The concentration is approximately 50 L / mol, and in some embodiments, it is 5 to 20 L / mol.

[0402] In some embodiments, the cyclic acid anhydride is succinic acid anhydride, glutaric acid anhydride. It is one of the substances, adipic anhydride, or pimeric acid anhydride, and in some embodiments , succinic anhydride. The cyclic acid anhydride and the compound represented by formula (313) The ratio is 1:1 to 10:1, and in some embodiments, it is 2:1 to 5:1.

[0403] The esterification catalyst may be any catalyst that catalyzes the esterification reaction, for example. For example, the catalyst may be 4-dimethylaminopyridine. The catalyst and formula (313 The molar ratio with the compound shown in ) is 1:1 to 10:1, and in some embodiments The ratio is 2:1 to 5:1.

[0404] In some embodiments, the base is any inorganic base, organic base, or combination thereof. They may be combined. Considering solubility and product stability, the base may be, for example, the It may be a tertiary amine. In some embodiments, the tertiary amine is a tri- It is ethylamine or N,N-diisopropylethylamine. The above tertiary amine and formula (3 The molar ratio with the compound shown in 13) is 1:1 to 20:1, and in some embodiments... And the ratio is 3:1 to 10:1.

[0405] The aforementioned ion exchange action involves transferring the compound of formula (321) to a desired carboxylic acid or carboxylate. The process involves converting to a specific format, and the ion exchange method is known to those skilled in the art, and appropriate ion exchange Using the solution and exchange conditions, M + A complex molecule containing a cation can be obtained, and here we will explain in detail. The explanation is omitted. In some embodiments, the ion exchange reaction is performed by triethyl The procedure is performed using a triethylamine phosphate solution, with a concentration of 0.2- The concentration is 0.8M, and in some embodiments, the triethylamine phosphate solution The concentration is 0.4 to 0.6 M, and the triethylamine is used with respect to the compound of formula (313). The dosage of the phosphate solution is 3-6 L / mol, and in further embodiments, 4-5 L / m². I am an office lady.

[0406] To isolate the compound of formula (321) from the reaction mixture by any suitable isolation method. This can be done. In some embodiments, after the solvent is evaporated and removed, chromatography is performed. The compound of formula (321) can be isolated by the method, for example, (1) normal-phase purified silicone : 200-300 mesh silica gel packing material containing 1 wt‰ triethylamine Chloromethane:methanol = 100:18~100:20 gradient elution, or ( 2) Reverse-phase purification: C18, C8 reverse-phase packing material, methanol:acetonitrile = 0.1:1 Isolation can be achieved using two chromatography conditions: gradient elution at a ratio of 1:0.1. In some embodiments, the solvent is directly removed to obtain the crude product of the compound of formula (321). This can be obtained, and the crude product can be used directly in subsequent reactions.

[0407] In some embodiments, the method for preparing the compound of formula (321) is under condensation reaction conditions. In an organic solvent, in the presence of a condensing agent, a condensation catalyst, and a tertiary amine, the above ion exchange reaction The product obtained by the response is further brought into contact with a solid support containing an amino group or a hydroxyl group. This further includes the following: In this case, R4 includes a first functional group and a second functional group, and the first A functional group containing a hydroxy protecting group, and a second functional group containing the structure shown in formula (C1'). Compound (321) is obtained.

[0408] The aforementioned solid-phase support is one of the supports used in the solid-phase synthesis of siRNA, and among them The details are known to those skilled in the art. For example, the solid support is an activated hydroxyl group or an amino A solid support containing functional groups may be selected, and in some embodiments, the solid support The body is an amino resin or a hydroxy resin. In some embodiments, the amino The resin or hydroxy resin has a particle size of 100 to 400 mesh, and on the surface The parameter for the amount of amino or hydroxyl group supported is 0.2 to 0.5 mmol / g. The dose ratio of the compound shown in formula (321) to the solid support is 10 to 400 μmol. The solid phase support is a compound per gram (μmol / g). In some embodiments, the above The dose ratio of the compound shown in formula (321) to the solid support is 50 to 200 μmol / g. .

[0409] The aforementioned organic solvent may be any suitable solvent or mixture known to those skilled in the art. In several embodiments, the organic solvent is acetonitrile, epoxy solvents, ether Solvents, alkyl halides, dimethyl sulfoxide, N,N-dimethylformaldehyde It is one or more of mido and N,N-diisopropylethylamine. Several implementations In form, the epoxy solvent is dioxane and / or tetrahydrofuran. The ether solvent is ethyl ether and / or methyl tert-butyl ether. Yes, the alkyl halogenated solvents are dichloromethane, trichloromethane and 1,2 - One or more types of dichloroethane. In some embodiments, the organic solvent This is acetonitrile. For the compound of formula (321), the amount of the organic solvent is 20 It is ~200 L / mol, and in some embodiments, it is 50~100 L / mol. ru.

[0410] In some embodiments, the condensing agent is benzotriazole-1-yloxy Tripyrrolidino-phosphonium hexafluorophosphate / ester (benzotr iazol-1-yl-oxytripyrrolidino phosphonium hexafluorophosphate (PyBop), 3-diethoxyphosphoryl -1,2,3-Benzoxazole 4(3H)-one (3-(Diethoxyphos phoryloxy)-1,2,3-benzotriazin-4(3H)-one, DEPBT) and / or O-benzotriazole-tetramethyluronium hexafluor Lophosphate / ester (O-benzotriazol-1-yl-tetrame It may also be thyluronium hexafluorophosphate. In some embodiments, the condensing agent is O-benzotriazole-tetramethyl It is rhonium hexafluorophosphate / ester. The condensing agent is shown in formula (321). The molar ratio with the compound being used is 1:1 to 20:1, and in other embodiments it is 1:1 to 5: It is 1.

[0411] In some embodiments, the tertiary amine is triethylamine and / or N, It is N-diisopropylethylamine, and in some embodiments, N,N-diiso It is a propylethylamine, and is a compound of the tertiary amine and the compound shown in formula (321). The ratio is 1:1 to 20:1, and in some embodiments, it is 1:1 to 5:1.

[0412] In some embodiments, the method for preparing the compound of formula (321) involves a capping reaction. Under these conditions, the obtained condensation product is treated with a capping reagent and an acylation catalyst in an organic solvent. This may further include contacting the compound with another compound and isolating it to obtain the compound shown in formula (321). The capping reaction prevents the formation of unwanted by-products in subsequent reactions. To do this, the first step is to remove any active functional groups that have not yet fully reacted. The conditions for the capping reaction are a reaction temperature of 0-50°C, and in some embodiments, 15-3°C. The temperature is 5°C, and the reaction time is 1 to 10 hours, or 3 to 6 hours in some embodiments. The capping reagent used is a capping reagent known to those skilled in the art and used for siRNA solid-phase synthesis. That's fine.

[0413] In some embodiments, the capping reagent is capping reagent 1 (cap 1) consists of capping reagent 2 (cap2), and capping reagent 1 is N-methyl It is an imidazole, and in some embodiments, a pyridine of N-methylimidazole. / Provided as an acetonitrile mixed solution, with a volume ratio of pyridine to acetonitrile of 1: The ratio is 10 to 1:1, and in some embodiments, 1:3 to 1:1, with pyridine and The volume ratio of acetonitrile to N-methylimidazole is 1:1 to 10:1. In some embodiments, the ratio is 3:1 to 7:1. The capping reagent 2 is vinegar. It is an acid anhydride. In some embodiments, the capping reagent 2 is an acetic anhydride. It is provided as an acetonitrile solution, with a volume ratio of acetic anhydride to acetonitrile of 1:1 The ratio is approximately 1:10, and in further embodiments, it is 1:2 to 1:6.

[0414] In some embodiments, the pyridine / acetonite of the N-methylimidazole The ratio of the volume of the mixed solution to the mass of the compound in formula (321) is between 5 ml / g and 50 ml / g. In some embodiments, the concentration is 15 ml / g to 30 ml / g. The acetate anhydride of the acetic acid. The ratio of the volume of the tonitrile solution to the mass of the compound in formula (321) is 0.5 ml / g to 10 ml. The concentration is / g, and in some embodiments it is 1ml / g to 5ml / g.

[0415] In some embodiments, equimolar amounts of anhydride acetate and N2 are used as the capping reagent. -Methylimidazole is used. In some embodiments, the organic solvent is acetone. Nitriles, epoxy solvents, ether solvents, alkyl halogens, dimethyl sulfur One of the following: hooxide, N,N-dimethylformamide and N,N-diisopropylethylamine It is one or more species. In some embodiments, the organic solvent is acetonitrile. Yes. For the compound of formula (321), the amount of the organic solvent is 10 to 50 L / mol. In some embodiments, the concentration is 5 to 30 L / mol.

[0416] In some embodiments, the acylation catalyst undergoes esterification condensation or amidation condensation. Any catalyst that can be used may be selected, for example, from alkali heterocyclic compounds. In this embodiment, the acylation catalyst is 4-dimethylaminopyridine. The mass ratio of the catalyst to the compound shown in formula (321) is 0.001:1 to 1:1, and how many In that embodiment, the ratio is 0.01:1 to 0.1:1.

[0417] In some embodiments, the reaction mixture is isolated from formula (321) by any suitable isolation method. The compound can be isolated. In some embodiments, it is thoroughly washed with an organic solvent. Purify, filter, and remove unreacted reactants, excess capping reagents, and other impurities. By this, a compound of formula (321) can be obtained. The organic solvent is acetonitrile, Selected from dichloromethane and methanol, in some embodiments, acetonitrile It is.

[0418] In some embodiments, the method for preparing the complex molecule represented by formula (321) is an organic solvent In the agent, under coupling reaction conditions and in the presence of a coupling reagent, formula (313) The compound shown was contacted with phosphordiamidite, isolated, and chemically converted as shown in formula (321). This includes obtaining a compound. In this case, R4 contains a first functional group and a second functional group, and the first The functional group of the first is a hydroxy protecting group, and the second functional group is a formula containing the structure shown in formula (C3). Compound (321) is obtained.

[0419] In some embodiments, the coupling reaction conditions are such that the temperature is 0 to 50°C. Often, for example, the temperature range is 15-35°C. The compound of formula (313) and phosphordiamidite The ratio can be anywhere from 1:1 to 1:50, for example, 1:5 to 1:15, as shown in equation (313). The molar ratio of the compound to the coupling reagent may be 1:1 to 1:100, for example 1 The reaction time is 50 to 1:80. The reaction time may also be 200 to 3000 seconds, for example, 500 The duration is approximately 1500 seconds. The phosphordiamidite is, for example, bis(diisopropylamino (2-cyanoethoxy)phosphine may be used, or a commercially available product may be purchased, or this It may be synthesized by methods known in the field. The coupling reagent is 1H-tetrazole. Selected from 5-ethylthio-1H-tetrazole and 5-benzylthio-1H-tetrazole It is one or more types, for example, 5-ethylthio-1H-tetrazole. The pulling reaction may be carried out in an organic solvent, the organic solvent being anhydrous acetonitrile, One or more substances selected from water DMF and anhydrous dichloromethane, for example, anhydrous acetone. It is a nitrile. In some embodiments, the compound of formula (313) is given the aforementioned The solvent concentration is 3 to 50 L / mol, or for example, 5 to 20 L / mol. By performing this coupling reaction, the hydroxyl group in the compound of formula (313) The phosphoramidite is reacted with a phosphoramidite to form a phosphoramidite group. Several implementations In terms of form, the solvent can be directly removed to obtain a crude product of the compound of formula (321), The crude product can be used directly in subsequent reactions.

[0420] In some embodiments, the method for preparing the compound of formula (321) involves a coupling reaction. Under these conditions, in an organic solvent, in the presence of a coupling reagent, the isolated product was obtained The process further includes contacting the hydroxyl group-containing solid support. Subsequently, the capping The compound of formula (321) is obtained by performing a oxidative reaction and then isolating it. In this case, R4 is the first It contains a functional group and a second functional group, the first functional group contains a hydroxy protecting group and the second functional group A compound of formula (321) is obtained in which the active group has the structure shown in formula (C3').

[0421] In some embodiments, the solid support is used in solid-phase nucleic acid synthesis known in the art. It is a solid phase support that can be used, for example, a commercially available general-purpose solid phase support that has undergone a deprotection reaction (NittoPh ase(registered trademark)HL UnyLinker TM 300 Oligonucleot ide Synthesis Support, Kinovate Life Science The structure may also be as shown in formula B80 (nces Inc.).

[0422] [ka]

[0423] The deprotection reaction is known to those skilled in the art. In some embodiments, the deprotection conditions are: The temperature range is 0-50°C, for example, 15-35°C, and the reaction time is 30-300 seconds, for example, 50- The time is 150 seconds. The deprotection reagents are trifluoroacetic acid, trichloroacetic acid, dichloroacetic acid, and It may be one or more selected from loroacetic acid, and in some embodiments, The deprotection agent is dichloroacetic acid. The deprotection agent and the stationary phase are -DMTr(4,4' The molar ratio with the (dimethoxytrityl) protecting group is 2:1 to 100:1, for example, 3:1 to The ratio is 50:1. By performing the deprotection, the surface of the solid support has reactive activity. A free hydroxyl group is obtained, facilitating the subsequent coupling reaction.

[0424] The coupling reaction conditions and the selection of coupling reagents are as described above. By performing the pulling reaction, the free hydroxyl group formed in the deprotection reaction and the phosphorus The midite group reacts to form a phosphite ester bond.

[0425] In some embodiments, the capping reaction conditions are such that the temperature is 0 to 50°C, for example 1 The temperature is 5-35°C, the reaction time is 5-500 seconds, for example, 10-100 seconds, and the aforementioned cap The Ping reaction is performed in the presence of a capping reagent. The selection and dosage of the capping reagent are as follows: That is correct.

[0426] The oxidation reaction conditions may be a temperature of 0 to 50°C, for example, 15 to 35°C, and the reaction time The incubation period can be 1 to 100 seconds, or even 5 to 50 seconds, and the oxidizing agent can be, for example, iodine. Good (provided as iodine water in some embodiments). In some embodiments, acid The molar ratio of the chemical reagent to the nucleic acid sequence bound to the solid support is 1:1 to 100:1, for example. The ratio may be 5:1 to 50:1. In some embodiments, the oxidation reaction is tetra This is carried out using a mixed solvent of hydrofuran:water:pyridine = 3:1:1 to 1:1:3.

[0427] In some embodiments, R6 is one of the bases of formula B7 or B8.

[0428] [ka] In the formula, the definition of q2 is as described above.

[0429] In this case, the compound shown in formula (313) reacts in an organic solvent under amidation reaction conditions. And in the presence of an amidation reaction condensing agent and a tertiary amine, the compound shown in formula (314) is converted to formula The compound shown in (A-1) or the compound of formula (A-2) is brought into contact with the compound, and then isolated. It can be obtained by the following preparation method.

[0430] [ka] In the formula, n1, n3, m1, m2, m3, R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , L1, S1, q2 and R k The definitions and selectable ranges for each are as described above. be.

[0431] The amidation reaction conditions are a reaction temperature of 0 to 100°C and a reaction time of 1 to 48 hours. In some embodiments, the amidation reaction conditions are such that the reaction temperature is 10-40°C. The reaction time is 2 to 16 hours.

[0432] In some embodiments, the organic solvent is an alcohol solvent, an epoxy solvent, Ether solvents, alkyl halogen solvents, dimethyl sulfoxide, N,N-dimethyl It is one or more of formamide and N,N-diisopropylethylamine. In some embodiments, the ethanol solvent is methanol, ethanol, propano It is one or more types of , and in some embodiments, it is ethanol. In some embodiments, the xylic solvent is dioxane and / or tetrahydrofuran. The ether solvent is, in some embodiments, ethyl ether and / or It is methyl tert-butyl ether. The alkyl halogen solvents are several In the embodiment, dichloromethane, trichloromethane and 1,2-dichloroethane It is one or more types. In some embodiments, the organic solvent is dichloromethane. Therefore, for the compound of formula (314), the amount of organic solvent is 3 to 50 L / mol. In further embodiments, the concentration is 3 to 20 L / mol.

[0433] In some embodiments, the amidation reaction condensation agent is benzotriazole-1- Il-oxytripyrrolidino-phosphonium hexafluorophosphate / ester, 3 -diethoxyphosphoryl-1,2,3-benzoxazole 4(3H)-one, 4-(4 ,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (4-(4,6 -dimethoxytriazin-2-yl)-4-methylmorpholi (1,2 hydrochloride), 2-ethoxy-1-ethoxycarbonyl-1,2 -Dihydroquinoline (EEDQ) or O-benzotriazole-tetramethyluronium Hexafluorophosphate / ester, and in further embodiments, 3-diethoxy It is cyphosphoryl-1,2,3-benzoxazole 4(3H)-one. The amidation The molar ratio of the reaction condenser to the compound shown in formula (314) may be 1:1 to 10:1. In some embodiments, the ratio is 2.5:1 to 5:1.

[0434] In some embodiments, the tertiary amine is triethylamine or N,N-di It is isopropylethylamine, and in further embodiments, N,N-diisopropyl ethylamine It is a thylamine. The molar ratio of the tertiary amine to the compound shown in formula (314) is 3: The ratio is 1 to 20:1, and in some embodiments, it is 5 to 10:1.

[0435] In some embodiments, compounds of formulas (A-1) and (A-2) are used as appropriate. It may be prepared by any method. For example, R k If it is a DMTr group, glyceryl phosphate The compound of formula (A-1) can be prepared by reacting Calcium with DMTrCl. As shown, 3-amino-1,2-propanediol and a cyclic acid anhydride are brought into contact, and then DMTr The compound of formula (A-2) can be prepared by reacting it with Cl, and the cyclic acid anhydride is carbon The cyclic acid anhydride may have 4 to 13 primary atoms, or in some embodiments, 4 to 8. As can be easily understood by those skilled in the art, the selection of the cyclic acid anhydride is the combination of (A-2) It corresponds to different values ​​of q2 in the substance, for example, the cyclic acid anhydride is succinic acid anhydride If so, q2=1, and if the cyclic acid anhydride is glutaric acid anhydride, q2= The answer is 2, and by analogy, this is also correct.

[0436] In some variations, the compound shown in formula (314) is the cyclic acid anhydride, 3- By reacting amino-1,2-propanediol and DMTrCl in sequence, the formula ( Compound 313) can also be prepared. As will be easily understood by those skilled in the art, These modifications do not affect the structure and function of the compound of formula (313), and This can be easily achieved by the contractor using the method described above.

[0437] Similarly to the above, the compound of formula (313) can be isolated from the reaction mixture by any suitable isolation method. It can be separated. In some embodiments, after the solvent is evaporated and removed, chromatography The compound of formula (313) can be isolated by the Raffie method, for example, (1) normal phase pulverization. Silica gel: 200-300 mesh silica gel packing material, petroleum ether: ethyl acetate L:Dichloromethane:N,N-dimethylformamide = 1:1:1:0.5~1:1:1 (2) Gradient elution at 0.6, and (2) Reverse phase purification: C18, C8 reverse phase packing material, Two characteristics of the compound are that it undergoes gradient elution at a ratio of 0.1:1 to 1:0.1 for acetonenitrile. It can be isolated under matrixing conditions. In some embodiments, the solvent can be directly By removing it, a crude product of the compound of formula (313) can be obtained, and this crude product can be used as is. It can be used in reactions.

[0438] In some embodiments, the compound represented by formula (314) is used in organic solvents. Under condensation reaction conditions in the presence of an amidation reaction coupling agent and a tertiary amine, the reaction shown in formula (320) A preparation method comprising contacting a compound with a compound represented by formula (316) and then isolating it. It can be obtained by law.

[0439] [ka] In the formula, n1, n3, m1, m2, m3, R 10 , R 11 , R 12 , R 13 , R 14 , R 15 The definitions and selectable ranges for each are as described above.

[0440] The compound of formula (316) is, for example, described in J. Am. Chem. Soc. 2014. Compounds disclosed in 136, 16958-16961 may be used, or formula (31 6) The compound can be prepared by various methods by those skilled in the art, for example, U.S. Patent Refer to the method disclosed in Example 1 of B2, and several formulas (316 The compound can be prepared, and by citation, all the contents of the above literature are included in this specification. It will be incorporated into the book.

[0441] In some embodiments, the condensation reaction conditions are such that the reaction temperature is 0 to 100°C, and the reaction time The interval is 0.1 to 24 hours, and in some embodiments, the reaction temperature is 10 to 40°C. The duration is between 0.5 and 16 hours.

[0442] Considering the structure of the compound of formula (314), which is the desired product, as shown in formula (316) The molar ratio of the compound to be used and the compound shown in formula (320) is given by formula (320) It should be determined based on the sum of n1 and n3. In some embodiments, for example, In the case where n1 + n3 = 3, in order to ensure that the reaction is complete and not in excess, the equation ( The molar ratio of the compound shown in formula (316) to the compound shown in formula (320) is 3:1 to 3 It may be 0.5:1, and in some embodiments, 3.01:1 to 3.15:1 be.

[0443] In some embodiments, the organic solvent is acetonitrile, epoxy solvents, etc. methyl solvents, alkyl halogens, dimethyl sulfoxide, N,N-dimethyl hydroxypropyl alcohol The epoxy is one or more of the following: lumamide and N,N-diisopropylethylamine. In some embodiments, the xylic solvent is dioxane and / or tetrahydrofuran. The ether solvent is, in some embodiments, ethyl ether and / or It is methyl tert-butyl ether, and the alkyl halogen solvent is several In the embodiment, dichloromethane, trichloromethane and 1,2-dichloroethane There may be one or more types, and in some embodiments, the organic solvent is dichloromethane. Therefore, for the compound of formula (320), the dose of the organic solvent is 3 to 50 L / mol. In some embodiments, the concentration is 5 to 20 L / mol.

[0444] In some embodiments, the amidation reaction condensation agent is benzotriazole-1- Il-oxytripyrrolidino-phosphonium hexafluorophosphate / ester, 3 -Diethoxyphosphoryl-1,2,3-benzoxazole 4(3H)-one (DEPB T), O-benzotriazole-tetramethyluronium hexafluorophosphate / Ester, 4-(4,6-dimethoxytriazin-2-yl)-4-methylmorpholine salt The salt is one or more of the salt or 1-hydroxybenzotriazole, and in further embodiments... In this case, benzotriazole-1-yl-oxytripyrrolidino-phosphonium hexaf It is a mixture of ruolophosphate / ester and 1-hydroxybenzotriazole, Nzotriazole-1-yl-oxytripyrrolidino-phosphonium hexafluorophosphonium Equimolar amounts of phosphate / ester and 1-hydroxybenzotriazole are used. The molar ratio of the total amidation reaction condenser to the compound shown in formula (316) is 1:1 to 3:1. It is also possible that the ratio is 1.05:1 to 1.5:1 in some embodiments.

[0445] The tertiary amine is N-methylmorpholine, triethylamine, or N,N-diisopropylamine. It may also be propylethylamine, and in some embodiments, N-methylmorphol The tertiary amine and the compound represented by formula (316) are in a molar ratio of 2:1 to 10. The ratio may be 1, and in some embodiments, it is 2:1 to 5:1.

[0446] Similarly to the above, the compound of formula (314) can be isolated from the reaction mixture by any suitable isolation method. It can be separated. In some embodiments, after the solvent is evaporated and removed, chromatography The compound of formula (314) can be isolated by the Raffie method, for example, (1) normal phase pulverization. Silica gel: 200-300 mesh silica gel packing material, dichloromethane: methyl (2) Reverse phase purification: C18, C 8. Elute the reversed-phase packing material with a gradient elution of methanol:acetonitrile = 0.1:1 to 1:0.1. It can be isolated under two chromatographic conditions. Several embodiments In this process, the solvent can be directly removed to obtain a crude product of the compound of formula (314), and the crude The product can be used directly in subsequent reactions.

[0447] Compound (320) can be obtained by purchasing a commercially available product or by methods known to those skilled in the art. This is possible. For example, m1=m2=m3=3, n1=1, n3=2, and each R 10 , R 11 , R 12 , R 13 , R 14 , R 15 If all of them are H, then the compound of formula (320) It can be purchased commercially from Alpha Acer Corporation.

[0448] The siRNA complex of this disclosure may be used in combination with other pharmaceutically acceptable additives. The additive may be one or more of the various formulations or compounds commonly used in this field. For further details, please refer to the description of the drug composition in this disclosure above.

[0449] <Use of the siRNA, drug composition and siRNA complex of this disclosure> In some embodiments, the Disclosure relates to the siRNA and / or drug compositions of the Disclosure. and / or siRNA complex, uric acid metabolism disorder or disease or physiological condition due to uric acid metabolism disorder Provides use in the preparation of drugs for the treatment and / or prevention of [condition]. Several embodiments In this context, the disease or physiological condition resulting from the uric acid metabolism disorder is hyperuricemia or gout.

[0450] In some embodiments, the Disclosure relates to the siRNA and / or drug compositions of the Disclosure. and / or administering an effective amount of the siRNA complex to a subject requiring it. , propose methods for the prevention and / or treatment of uric acid metabolism disorders or diseases or physiological conditions caused by uric acid metabolism disorders. Provided. In some embodiments, the disease or physiological condition due to the uric acid metabolism disorder is high The patient has uricemia or gout.

[0451] By administering the siRNA active component of this disclosure to a subject requiring it, RN A interference mechanism prevents uric acid metabolism disorders or diseases or physiological conditions caused by uric acid metabolism disorders and / or Therefore, the siRNAs and / or siRNAs of this disclosure can achieve the therapeutic purpose. The drug composition and / or siRNA complex is for uric acid metabolism disorders or diseases caused by uric acid metabolism disorders. It is used for the prevention and / or treatment of physiological conditions, or for uric acid metabolism disorders or conditions caused by uric acid metabolism disorders. It can be used in the preparation of drugs for the prevention and / or treatment of diseases or physiological conditions. In several embodiments, the uric acid metabolic disorder or the disease or physiological condition caused by the uric acid metabolic disorder is , hyperuricemia or gout.

[0452] As used herein, the term "drug administration / administration" refers to the siRNA, drug composition of this disclosure. and / or localize at least a portion of the siRNA complex to a desired site to produce the desired effect. By a method or route that causes the siRNA, drug composition and / or siRNA polymorphism of the Disclosure, This refers to introducing the combined substance into the body of the subject. Suitable administration routes for the method disclosed herein include local administration and This includes systemic administration. Generally, local administration allows for a greater amount of siRNA to be administered than is available in the subject's systemic circulation. The complex is delivered to a specific site, but systemic administration allows the siRNA, drug composition of this disclosure to be used. and / or the siRNA complex is delivered to the subject's basic systemic circulation. This disclosure relates to uric acid metabolism. To provide means for the prevention and / or treatment of diseases or physiological conditions caused by abnormalities or uric acid metabolism disorders. Considering the intent, in some embodiments, the drug is delivered to the liver. We will adopt an administration method that allows for this.

[0453] The subject can be administered via any appropriate route known in this field, such as: Oral administration or extra-gastrointestinal routes, such as intravenous, intramuscular, subcutaneous, transdermal, or tracheal administration. Intravenous administration (aerosol), pulmonary administration, nasal administration, rectal administration, and local administration (oral administration and sublingual administration). This includes, but is not limited to, administration of, daily, weekly, or bi-weekly doses. This may be done once or multiple times every three weeks, once a month, or once a year.

[0454] The doses of siRNA, drug compositions, or siRNA complexes described herein are in the Art of the Disclosure. The usual dose may be used, and the dose may be determined by various parameters, particularly the age of the subject. It may also be determined by weight and sex. Toxin can be obtained in cell cultures or experimental animals using standard pharmaceutical procedures. Sexuality and therapeutic effect are measured, for example, LD 50 (Dose that causes death in 50% of the population) and ED5 0 (In quantitative reactions, this refers to the dose that can produce 50% of the maximum response intensity.) (In qualitative reactions, this refers to the dose at which a positive reaction occurs in 50% of the experimental subjects.) This may be done. Based on data obtained from cell culture analysis and animal studies, the range of human doses may be determined. You can obtain an enclosure.

[0455] Administering the siRNA, drug composition, and / or siRNA complex described herein. For example, a male or female, 6-12 weeks old, weighing 18-25g C57BL / 6J or For 30-45g of ob / ob mice, the amount of siRNA is (i) multiple siRNAs. Regarding the combination, the siRNA dose may be 0.001 to 100 mg / kg body weight. In some embodiments, the amount is 0.01 to 50 mg / kg body weight, and in some embodiments... The amount is 0.05 to 20 mg / kg body weight, and in some embodiments, it is 0.1 to The dosage is 15 mg / kg body weight, and in some embodiments, it is 0.1 to 10 mg / kg (ii) a drug combination formed by (ii) siRNA and a pharmaceutically acceptable carrier, with a weight of g. Regarding the product, the siRNA dose may be 0.001 to 50 mg / kg body weight. In some embodiments, it is 0.01 to 10 mg / kg body weight, and in some embodiments The concentration is 0.05 to 5 mg / kg body weight, and in some embodiments, it is 0.1 to 3 mg / kg body weight. That is the case.

[0456] In some embodiments, the Disclosure relates to the siRNA and / or drug compositions of the Disclosure. and / or bring an effective amount of the siRNA complex into contact with the cells, and the siRNA of the Disclosure, and / or The drug composition and / or siRNA complex is introduced into the cells, and the RNA interference mechanism is used to further refine the cells. This includes achieving the objective of suppressing the expression of the XO gene in the cell, The present invention provides a method for suppressing the expression of the O gene. The cells are SMMC-7721 and CAL-27. They may be selected from cancer cell lines such as Huh7 or isolated primary hepatocytes. In the application morphology, the cells are CAL-27 cells.

[0457] The method provided herein suppresses the expression of the XO gene in cells. The siRNA dose in the modified siRNA, drug composition and / or siRNA complex is Generally, it can reduce the expression of target genes, reaching levels of 1 pM to 1 μM, or 0.0, on the surface of target cells. Fine particles ranging from 0.1nM to 100nM, or 0.05nM to 50nM, or 0.05nM to approximately 5nM. This is the amount that results in the extracellular concentration. The amount required to achieve the local concentration depends on the delivery method and the delivery site. This includes the number of cell layers between the delivery site and the target cells or tissue, and the delivery route (local or systemic), etc. It changes due to various factors. The concentration at the delivery site is on the surface of the target cells or tissue. The concentration may be significantly higher than the specified concentration.

[0458] <Kit> This disclosure relates to at least one of the modified siRNAs, drug compositions, and siRNA complexes of this disclosure. We provide a kit containing an effective amount of seeds.

[0459] In some embodiments, the kit described herein includes a container for modified siRNA. It is possible to provide. In some embodiments, the kit described herein is The container may include a container that provides pharmaceutically acceptable excipients. Furthermore, the aforementioned kit may contain other ingredients, such as stabilizers or preservatives. In several embodiments, the kit described herein is modified according to the specifications described herein. The container providing the iRNA may contain at least one other therapeutic agent in a separate container. In several embodiments, the kit comprises modified siRNA and a pharmaceutically acceptable carrier. It may also include instructions for mixing in and / or additives or other ingredients (if any).

[0460] In the kit of this disclosure, the modified siRNA and a pharmaceutically acceptable carrier and / or Additives, and the modified siRNA, drug composition and / or siRNA complex, and / Alternatively, pharmaceutically acceptable additives may be in any form, e.g., liquid form, dry form, or freeze-dried form. It may be provided in a dry form. In some embodiments, the modified siRNA and A pharmaceutically acceptable carrier and / or additive, and the drug composition and / or siR NA complexes and any pharmaceutically acceptable additives are essentially clean and / or sterile. Yes. In some embodiments, sterile water can be provided in the kit of this disclosure.

[0461] The present disclosure will be further illustrated below by examples, but the present disclosure is not limited in any way. do not have. [Examples]

[0462] Unless otherwise specified, the reagents and culture media used in the following examples are all commercially available products. The nucleic acid electrophoresis and real-time PCR procedures used are all performed by Mole. cular Cloning(Cold Spring Harbor Laborat This is done by referring to the method described in Ory Press (1989).

[0463] C57BL / 6N mice: 6-8 weeks old, purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd. Hereafter, this will simply be referred to as the C57 mouse.

[0464] Unless otherwise specified, all reagent ratios provided below are expressed as volume ratios (v / v). It will be calculated.

[0465] Unless otherwise specified, the experimental data on in-vitro / ex-vitro effects below are all

number

[0466] (Preparation Example 1) Preparation of siRNA complex L10-siXOi1M1S

[0467] In this preparation example, the siRNA complex L10-siXOi1M1S was synthesized. The NA complex is formed after the L-9 complex molecule is combined with the siRNA no. siXOi1M1S. This is the formed siRNA complex. The sequence of the siRNA complexed into this siRNA complex. This is shown in Table 3.

[0468] (1-1) Synthesis of L-10 compounds The L-10 compound was synthesized according to the following method. [ka]

[0469] (1-1-1) Synthesis of the composite terminal segment GAL-5 [ka]

[0470] (1-1-1a) Synthesis of GAL-2 100.0g of GAL-1 (N-acetyl-D-galactosamine hydrochloride, CAS number: Purchased on March 8, 1772, from Ningbo Hongxiang Biochemical Company, 1000 ml of 463.8 mmol. Dissolve in anhydrous pyridine and dissolve in 540 ml of anhydrous acetic acid (purchased from Enox) in an ice bath. Add 5565.6 mmol) and stir at room temperature for 1.5 hours. The reaction solution was then placed in 10 L of ice water. Inject into, filter under reduced pressure and suction, wash the cake with 2L of ice water, and then ace until completely dissolved. Add a tonitrile / toluene mixed solvent (volume ratio of acetonitrile:toluene = 1:1), The solvent was evaporated to dryness, yielding 130.0 g of a white solid product, GAL-2.

[0471] (1-1-1b) Synthesis of GAL-3 The GAL-2 (35.1g, 90.0 mmol) obtained in step (1-1-1a) was divided into 21 Dissolve in 3 ml of anhydrous 1,2-dichloroethane and steep in an ice bath under nitrogen protection conditions for 24 hours. 0g TMSOTf (CAS number: 27607-77-8, purchased from Maclin, 10 8.0 mmol was added and the mixture was reacted overnight at room temperature.

[0472] Dilute the reaction solution by adding 400 ml of dichloromethane, filter it through diatomaceous earth, and add 1 L of saturated carbon Add an aqueous solution of sodium oxyhydrogen, stir uniformly, separate the organic phase, and collect the aqueous phase from dichloroethane. The extraction was performed twice with 300ml each time, combining the organic phases, and then 300ml of saturated solution was added for each extraction. Wash with sodium bicarbonate aqueous solution and 300 ml of saturated saline solution to separate the organic phase, and then anhydrous The mixture was dried with sodium sulfate, and the solvent was evaporated under reduced pressure to dryness, yielding 26.9 g of a pale yellow, viscous, syrup-like substance. Product GAL-3 was obtained.

[0473] (1-1-1c) Synthesis of GAL-4 The GAL-3 (26.9g, 81.7 mmol) obtained in step (1-1-1b) was divided into 13 Dissolve in 6 ml of anhydrous 1,2-dichloroethane, add 30 g of dried 4 Å molecular sieve powder, 9.0g of 5-hexen-1-ol (CAS number: 821-41-0, Adamas- Purchased from Beta, add 89.9 mmol), stir at room temperature for 30 minutes, then ice bath and nitrogen Under protective conditions, add 9.08 g of TMSOTf (40.9 mmol) and stir overnight at room temperature. The 4Å molecular sieve powder was filtered out, and the filtrate was diluted with 300 ml of dichloromethane. Filter with diatomaceous earth, add 500 ml of saturated sodium bicarbonate solution, and stir for 10 minutes to wash. The solution is purified, the organic phase is separated, the aqueous phase is extracted once with 300 ml of dichloroethane, and the organic phase is foamed. Then, using 300 ml of saturated sodium bicarbonate solution and 300 ml of saturated saline solution, Wash, separate the organic phase, dry with anhydrous sodium sulfate, evaporate the solvent under reduced pressure to dryness, 41 0.3g of a yellow, syrup-like product, GAL-4, was obtained, and without purification, the following oxidation reaction was carried out. .

[0474] (1-1-1d) Synthesis of GAL-5 GAL-4 (14.9g, 34) obtained by the method described in step (1-1-1c) Dissolve 0.7 mmol) in a mixed solvent of 77 ml of dichloromethane and 77 ml of acetonitrile. Dissolve the mixture in 103 ml of deionized water and 29.7 g of sodium periodate (CA). S number: 7790-28-5, purchased from Aladdin, with 138.8 mmol added. Stir in an ice bath for 10 minutes to remove ruthenium(III) chloride (CAS number: 14898-6 Add 7-0 (purchased from Energy, 238 mg, 1.145 mmol) and leave at room temperature. The reaction was carried out in the evening. 300 ml of water was added to the reaction solution to dilute and stir, and saturated sodium bicarbonate was added. In addition, the pH was adjusted to approximately 7.5, the organic phase was separated and discarded, and the aqueous phase was treated with dichloromethane once. The mixture was extracted three times using 200ml of liquid, and the organic phase was discarded. The pH of the aqueous phase was adjusted to approximately 3 using solid citric acid. Adjust the concentration, extract with dichloromethane three times at 200 ml each time, combine the organic phase, and remove the excess. Dry with sodium hydroxide sulfate, evaporate the solvent under reduced pressure to dryness, and obtain 6.85 g of white foamy solid product G. I obtained AL-5. 1H NMR (400 MHz,DMSO) δ 12.01 (b r,1H),7.83 (d,J = 9.2 Hz,1H),5.21 (d,J = 3.2 Hz,1H),4.96 (dd,J = 11.2,3.2 Hz,1H) ,4.49 (d,J = 8.4 Hz,1H),4.07-3.95 (m,3H) ,3.92-3.85 (m,1H),3.74-3.67 (m,1H),3.48- 3.39 (m,1H),2.20 (t,J = 6.8 Hz,2H),2.11 (s,3H),2.00 (s,3H),1.90 (s,3H),1.77 (s,3 H), 1.55-1.45 (m, 4H).

[0475] (1-1-2) Synthesis of L-8 [ka]

[0476] J-0 (9.886g, 52.5mmol, purchased from Alpha Acer) and process ( 1-1-1) GAL-5 obtained (72.819 g, 162.75 mmol, multiple lo Dissolve the combined product of the set in 525 ml of dichloromethane, and diisopropyl Ethylamine (DIEA, 44.782g, 346.50mmol), benzotriazole Lu-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate / es Tel (PyBOP, 90.158g, 173.25mmol) and hydroxybenzotri Add azole (HOBt, 23.410 g, 173.25 mmol) and react at room temperature for 4 hours. Then, wash by adding 20 ml of saturated sodium bicarbonate and 200 ml of saturated saline solution. The aqueous phase was extracted twice with dichloromethane at a rate of 100 ml each time, and the organic phase was combined with anhydrous sulfur. The crude product was obtained by drying with sodium acid, filtering, and then evaporating the solvent under reduced pressure to dryness. Using 200-300 mesh normal-phase silica gel, and 10 wt% triethylamine, Neutralize the acidity of Kagel, equilibrate the column with 1 wt‰ triethylamine, and dichloromethane Gradient elution was performed using methanol at a ratio of 100:25 to 100:40, and the product eluate was collected. The mixture was evaporated under reduced pressure to dryness to obtain 38.8 g of pure L-8. 1 1H NMR (400 MHz) ,DMSO) δ 7.84 (d,J = 9.0 Hz,3H),7.27-7.2 3 (m,1H),7.13-7.18 (m,1H),5.22 (d,J = 3. 1 Hz,3H),4.97 (dd,J = 11.3,3.1 Hz,3H),4. 48 (d,J = 8.4 Hz,3H),4.09-3.98 (m,9H),3. 88 (dd,J = 19.3,9.3 Hz,3H),3.75-3.66 (m, 3H),3.44-3.38 (m,3H),3.17-3.30 (m,4H),3. 10-2.97 (m,4H),2.35-2.20 (m,6H),2.15-2.0 8 (m,9H),2.07-1.98 (m,13H),1.94-1.87 (m, 9H),1.81-1.74 (m,9H),1.65-1.42 (m,18H). MS m / z:C 85 H 119 N7O 30 [M+H] + Theoretical value: 1477.59, Actual Measured value: 1477.23.

[0477] (1-1-3a) Synthesis of A-1 [ka]

[0478] DMTrCl(4,4'-bismethoxytrityl chloride, 101.65g, 300ml) Dissolve (mol) in 1000 ml of anhydrous pyridine and hydrate DL-glycerate calcium Add the substance (28.63g, 100 mmol), react at 45°C for 20 hours, and filter the reaction solution. Rinse the cake with 200 ml of DCM, concentrate the filtrate under reduced pressure until dry, and use 500 ml of the remainder. Dissolve again in 1 liter of dichloromethane and 0.5 M triethylamine phosphate (pH=7~ 8) Wash twice with 200 ml each time, and rinse the aqueous phase with dichloromethane, 20 ml each time. Extract twice with 0 ml, combine the organic phases, dry with anhydrous sodium sulfate, filter, and remove the solvent. The solution is evaporated to dryness under reduced pressure, purified using a normal-phase silica gel column with a 200-300 mesh size, and then processed into petroleum ether. L:ethyl acetate:dichloromethane:methanol = 1:1:1:0.35~1:1:1:0 Gradient elution was performed at 0.55, the product eluate was collected, the solvent was evaporated under reduced pressure to dryness, and 600 ml of the solution was added. Dissolve again in lolomethane and wash once with 200 ml of 0.5 M triethylamine phosphate. The solution is purified, the aqueous phase is extracted once with 200 ml of dichloromethane, and the organic phase is combined with anhydrous sodium sulfate. Dry with alum, filter, evaporate the solvent under reduced pressure to dryness, and then reduce pressure overnight with a vacuum oil pump for 50°C. 7 g of white solid product A-1 was obtained. 1 1H NMR (400 MHz, DMSO-d6) δ 7.46 (ddd,J = 6.5,2.3,1.1 Hz,1H),7.40 -7.28 (m,7H),6.89-6.81 (m,4H),4.84 (d,J = 5.0 Hz,1H),4.36-4.24 (m,1H),4.29 (s,6H ),3.92 (dd,J = 12.4,7.0 Hz,1H),3.67 (dd, J = 12.3,7.0 Hz,1H),2.52 (q,J = 6.3 Hz,6 H),1.03 (t,J = 6.3 Hz,9H).MS m / z:C 24 H 23 O6, [MH] - Theoretical value: 407.15, measured value: 406.92.

[0479] (1-1-3b) Synthesis of L-7 [ka]

[0480] L-8 obtained in process (1-1-2) (40g, 27.09 mmol, multiple lots) (combining the products) and A-1 obtained in process (1-1-3a) (41.418g, 81 Mix with (0.27 mmol) and dissolve in 271 ml of dichloromethane, then add 3-diethoxy Phosphoryl-1,2,3-benzoxazole 4(3H)-one (DEPBT) (24. Add 318g (81.37 mmol), and further add diisopropylethylamine (21.0 Add 0.7g (162.54 mmol) and stir at 25°C for 1.5 hours, then add 800ml The organic phase is washed with saturated sodium bicarbonate, and the aqueous phase is washed with dichloromethane, 50 ml per cycle. Extract three times with 1 ml, wash the organic phase with 150 ml of saturated saline solution, and remove the aqueous phase with 50 ml of dichloroethylene solution. Extract once with methane, combine with the organic phase, dry with anhydrous sodium sulfate, filter, and then dissolve. The agent was evaporated to dryness under reduced pressure, and then foam-dried overnight using a vacuum oil pump to obtain the crude product. For column purification... Using 2 kg of 200-300 mesh normal-phase silica gel, add 200 ml of triethyl alcohol. The acidity of the silica gel was neutralized with amine, and then colored with petroleum ether containing 1 wt% triethylamine. Equilibrate the mixture: petroleum ether: ethyl acetate: dichloromethane: N,N-dimethylformaldehyde Gradient elution was performed with a mid ratio of 1:1:1:0.5 to 1:1:1:0.6, and the product eluate was collected. The solvent was evaporated under reduced pressure to dryness to obtain 40.4 g of pure L-7. 1 1H NMR (400 M) Hz,DMSO) δ7.90-7.78 (m,4H),7.75-7.64 (m, 1H),7.38-7.18 (m,9H),6.91-6.83 (m,4H),5. 25-5.10 (m,4H),4.97 (dd,J = 11.2,3.2 Hz, 3H),4.48-4.30 (m,4H),4.02 (s,9H),3.93-3. 84 (m,3H),3.76-3.66 (m,9H),3.45-3.35 (m, 3H),3.24-2.98 (m,10H),2.30-2.20 (m,2H),2 .11-1.88 (m,31H),1.80-1.40 (m,28H). MS m / z:C 90 H 128 N7O 35 [M-DMTr] + Theoretical value: 1564.65, actual measurement Value: 1564.88.

[0481] (1-1-4) Synthesis of L-9 [ka]

[0482] L-7 (40g, 21.4247 mmol) obtained in process (1-1-3b), succinate Acid anhydride (4.288 g, 42.8494 mmol) and 4-dimethylaminopyridine ( Mix DMAP (5.235g, 42.8494mmol) in 215ml of dichloromethyl Dissolve in tan, and further diisopropylethylamine (DIEA, 13.845g, 10 Add 7.1235 mmol) and stir at 25°C for 24 hours, then add 800 ml of 0.5 M triethyl alcohol. Wash the reaction mixture with luminamine phosphate, and extract the aqueous phase with dichloromethane three times at a rate of 5 ml per extraction. The organic phase was added, and the crude product was obtained by evaporation under reduced pressure to dryness. For column purification, 20 oz (1 kg) was used. Using normal-phase silica gel of 0-300 mesh, 1 wt% triethylamine is used to... The acidity was neutralized, the column was equilibrated with dichloromethane, and a 1 wt‰ dichloromethane containing triethylamine was added. Gradient elution was performed using chloromethane:methanol = 100:18 to 100:20, and the product eluate was obtained. The mixture was recovered, and the solvent was evaporated to dryness under reduced pressure to obtain 31.0 g of pure L-9 composite molecules. 1 H NM R (400 MHz,DMSO) δ 8.58 (d,J = 4.2 Hz,1H ),7.94-7.82 (m,3H),7.41-7.29 (m,5H),7.22 (d,J = 8.1 Hz,5H),6.89 (d,J = 8.3 Hz,4H ),5.49-5.37 (m,1H),5.21 (d,J = 3.0 Hz,3H ),4.97 (d,J = 11.1 Hz,3H),4.49 (d,J = 8. 2 Hz,3H),4.02 (s,9H),3.88 (dd,J = 19.4,9 .4 Hz,3H),3.77-3.65 (m,9H),3.50-3.39 (m, 6H),3.11-2.90 (m,5H),2.61-2.54 (m,4H),2. 47-2.41 (m,2H),2.26-2.17 (m,2H),2.15-1.9 5 (m,22H),1.92-1.84 (m,9H),1.80-1.70 (m, 10H),1.65-1.35 (m,17H),1.31-1.19 (m,4H), 0.96 (t,J = 7.1 Hz,9H).MS m / z:C 94 H 132 N7 O 38 [M-DMTr] + Theoretical value: 1664.72, measured value: 1665.03.

[0483] (1-1-5) Synthesis of L-10 compounds [ka]

[0484] In this process, the L-9 composite molecule is bonded to the solid support, thereby forming the L-10 compound We prepared it.

[0485] L-9 composite molecule obtained in step (1-1-4) (22.751 g, 11 mmol), O -Benzotriazole-tetramethyluronium hexafluorophosphate / ester (HBTU, 6.257g, 16.5mmol) and diisopropylethylamine (DI Mix EA (2.843g, 22 mmol) and dissolve in 900ml of acetonitrile. Stir at room temperature for 5 minutes, and add aminomethyl resin (88g, 100-200 mesh) to the reaction solution. Add the amino group-supported material (400 μmol / g, purchased from Minamikai Kazunari Co., Ltd.) and shake at 25°C. The reaction was carried out using a DC filter, and after 18 hours of reaction at 150 revolutions per minute, the cake was filtered and the cake was DC filtered. Rinse twice with 300ml each time, then rinse with acetonitrile, 300ml each time. Rinse three times, dry for 18 hours using a vacuum oil pump, and then, according to the mixing ratio shown in Table 2, Ingredients (CapA, CapB, 4-dimethylaminopyridine (DMAP), and acetonitrile) The capping reaction was carried out by adding ( ). The mixture was left to stand in a shaker at 25°C and rotated at 150 rpm. React for 5 hours at a rate of / min, filter the reaction mixture, and rinse the cake with acetonitrile at a rate of 300 per batch. Rinse three times with ml, evaporate the solvent under reduced pressure until dry, and dry under reduced pressure overnight using a vacuum oil pump. 102 g of L-10 compound with a supported weight of 90.8 μmol / g (i.e., bonded to a solid support) We obtained an L-9 composite molecule.

[0486] [Table 2] Here, CapA and CapB are capping reagent solutions, and CapA is 20 volumes. This is a pyridine / acetonitrile mixture solution of % N-methylimidazole, and contains pyridine and a The volume ratio with cetonitrile is 3:5, and CapB is 20 vol% acetonitrile anhydride. This is Toll's solution.

[0487] (1-2) Synthesis of the sense strand of the siRNA complex L10-siXOi1M1S Starting with the L-10 compound prepared in the above step by the solid-phase phosphoramidite method The nucleotides are rotated and, following the sequence of nucleotides in the sense strand, one nucleocytose is added in each direction from 3' to 5'. A nucleoside monomer was attached. Each time a nucleoside monomer was attached, deprotection and coupling occurred. Four reactions were performed: capping, oxidation, or sulfidation. A phosphate group formed between two nucleotides. When bonded by sterol, when the next nucleoside monomer is bonded, deprotection and cut occur. Four reactions were performed: pulling, capping, and oxidation. Thiolin was used between two nucleotides. When bonded by an acid ester, when the following nucleoside monomers are bonded, protection, Four reactions were carried out: plucking, capping, and sulfidation. The synthesis conditions were defined as follows: did.

[0488] Nucleoside monomers are provided in a 0.1 M acetonitrile solution, and each deprotection reaction is performed. The conditions were kept the same, namely the temperature was 25°C and the reaction time was 70 seconds. The deprotection reagent was dichlorofluorocarbon. This is a 3% v / v solution of dichloroacetic acid in dichloromethane, and the 4 in dichloroacetic acid and solid support The molar ratio with the ,4'-dimethoxytrityl protecting group is 5:1.

[0489] The coupling reaction conditions were the same for all of them: the temperature was 25°C, and the coupling was performed on a solid support. The molar ratio of the nucleic acid sequence to the nucleoside monomer is 1:10, and the nucleic acid sequence is bound to the solid support. The molar ratio of the row to the coupling reagent was 1:65, and the reaction time was 600 seconds. Coupling The reagent is 5-(Ethylthio)-1H-tetrazole (5-(Ethylthio)-1H-t This is a 0.5 M acetonitrile solution of etrazole (ETT).

[0490] The capping conditions were the same for all cappings: a temperature of 25°C and a reaction time of 15 seconds. The capping reagent solution is a mixed solution of CapA and CapB in a molar ratio of 1:1. The molar ratio of the capping reagent to the nucleic acid sequence bound to the solid support is: anhydride acetate:N-methyl acetate Chilimidazole: The nucleic acid sequence bound to the solid support is 1:1:1.

[0491] The oxidation reaction conditions were the same for each reaction, with a temperature of 25°C and a reaction time of 15 seconds, and the oxidizing reagent was concentrated to the same concentration. This is 0.05 M iodine water. Iodine is bonded to a solid support during the coupling process. The molar ratio of the nucleic acid sequence is 30:1. The reaction is tetrahydrofuran:water:pyridine = 3 The procedure was performed using a 1:1 mixed solvent.

[0492] The conditions for each sulfidation reaction were the same: the temperature was 25°C, the reaction time was 300 seconds, and the sulfidation reagent was... It is xanthan hydride. It is bonded to a solid support in the coupling process with a sulfidation reagent. The molar ratio of the nucleic acid sequence is 120:1. The reaction is performed using acetonitrile:pyridine in a 1:1 ratio. It was done using a mixed solvent.

[0493] After the binding of the last nucleoside monomer is complete, the nucleic acid sequence is sequentially bound to the solid support. The sample was then cleaved, deprotected, purified, and desalted, and subsequently freeze-dried to obtain the sense chain.

[0494] The cleavage and deprotection conditions are as follows: The synthesized support is bound to the nucleotide. The column was added to 25 wt% ammonia solution, with a volume of ammonia solution of 0.5 ml / μmol. The mixture was reacted at 55°C for 16 hours, the remaining support was filtered off, and the supernatant was vacuum concentrated until dry. did.

[0495] The purification and desalting conditions are as follows: Preparative ion chromatography purification column. Nucleic acid purification was completed by gradient elution with NaCl using (Source 15Q). Specifically, the eluent A is 20 mM sodium phosphate (pH 8.1), and the solvent is water / acetone. The nitrile ratio is 9:1 (by volume), and the eluent B consists of 1.5M sodium chloride and 20mM phosphorus. The solution is sodium acetate (pH 8.1), and the solvent is water / acetonitrile in a volume ratio of 9:1. Elution gradient: Eluting agent A:Eluting agent B = 100:0 to 50:50. Product eluate was obtained. After recovery and combination, desalting was performed using a reverse-phase chromatography purification column, and specific conditions were determined. Then, desalting is performed using a dextran gel column, and the packing material is dextran gel G25 (Se It was identified as phadex G25 and eluted with deionized water.

[0496] The detection method is as follows: Ion exchange chromatography (IEX-HPLC) The purity of the sense chain is detected using the above method, and liquid chromatography-mass spectrometry (LC-MS) is performed. The molecular weight was analyzed using [method / method]. The theoretical value was 7584.5, and the measured value was 7584.0. The measured value matches the theoretical value, and the sense chain SS has an L-9 complex molecule compounded at its 3' end. This demonstrated that the synthesis was achieved.

[0497] (1-3) Synthesis of the antisense strand of the siRNA complex L10-siXOi1M1S The solid-phase phosphoramidite method allows for the production of general-purpose solid-phase supports (UnyLinker TM loa ded NittoPhase(registered trademark)HL Solid Supports, Ki Starting from Novate Life Sciences, the siRNA complex is recycled. Antisense chains of L10-siXOf1M1S were synthesized. Desorption was performed using a solid-phase synthesis method. Protection, coupling, capping, oxidation or sulfurization reaction conditions, cleavage and deprotection, purification and desalting The conditions were the same as for the synthesis of the sense chain. The antisense chain AS was obtained by freeze-drying.

[0498] The purity of the antisense chain was examined by ion exchange chromatography (IEX-HPLC). The molecular weight of the antisense chain was then determined by liquid chromatography-mass spectrometry (LC-MS). The analysis revealed that the measured values ​​matched the theoretical values, and the antisense strand possessed the target sequence. This demonstrated that AS was synthesized.

[0499] (1-4) Synthesis of siRNA complex L10-siXOi1M1S For the siRNA complex L10-siXOi1M1S, the sense strand and antisense strand are Each is dissolved in sterile water for injection to obtain a 40 mg / mL solution, which is then mixed in equimolar ratios and incubated at 50°C. After heating for 15 minutes and cooling to room temperature, the product is annealed and freeze-dried. Freeze-dried powder was obtained. Ultrapure water (Milli-Q ultrapure water apparatus, resistivity 18.2 MΩ*cm) After diluting the siRNA complex to a concentration of 0.2 mg / mL using 25°C, Liquid chromatography-mass spectrometry (LC-MS) aphy-Mass Spectrometry, purchased from Waters, model number: LC The molecular weight was detected using T Premier. The measured value was in agreement with the theoretical value, indicating that the synthesis was successful. The siRNA complex was determined to contain the target-designed double-stranded nucleic acid sequence, which includes the L-9 complex molecule. The structure is shown in formula (403). The siRNA is shown in Table 3. This sequence corresponds to the iRNA complex L10-siXOi1M1S.

[0500] [Table 3] However, uppercase C, G, U, and A represent the base sequence of a nucleotide, and lowercase m represents the same This indicates that the nucleotide adjacent to the left of the letter 'm' is a methoxy-modified nucleotide. Furthermore, the lowercase letter f is a fluoromodified nucleus, where one nucleotide adjacent to the left of the letter f is fluoromodified. The lowercase letter 's' indicates that there is a thio between the two nucleotides to the left and right of the letter 's'. This indicates that the groups are bonded by phosphate ester groups.

[0501] (Preparation Example 2 and Comparative Preparation Example 3) The sequence of the sense strand and antisense strand of the siRNA complex L10-siXOi1M1S In contrast, the sense chain and amplifier corresponding to L10-siXOk1M1S and NC in Table 3. Except for using a cysene chain, the preparation followed the method in Preparation Example 1, and further included the siRNA complex L10. -siXOk1M1S and comparative siRNA complex NC were synthesized. The siRNAs included in the composite are L10-siXOk1M1S and N in Table 3, respectively. It has a sense chain and an antisense chain corresponding to C.

[0502] After preparation is complete, the prepared siRNA complex L10-s is prepared according to the method of Preparation Example 1. When the molecular weights of iXOk1M1S and NC were detected, the measured values ​​were in agreement with the theoretical values. The synthesized siRNA complex is a target-design double-stranded nucleic acid sequence containing an L-9 complex molecule. This was shown. Their structures are shown in formula (403). These siRNA complexes The siRNAs included are the siRNA complexes L10-siXOk shown in Table 3. The sequence corresponded to 1M1S and NC.

[0503] (Preparation Examples 4-18 and Comparative Preparation Example 19) <Synthesis of siRNA provided in this disclosure> By solid-phase synthesis, the sense strand or antisense strand of the siRNA sequence shown in Table 4 is obtained. Each was synthesized, and using DEPC-treated water, equimolar complementary senses in Table 4 were obtained. After dissolving the chain and antisense chain separately, annealing is performed as shown in Table 4 of this disclosure. siXOa1M1S, siXOb1M1S, siXOc1M1S, si XOd1M1S, siXOe1M1S, siXOf1M1S, siXOg1M1S, si XOh1M1S, siXOi1M1S, siXOj1M1S, siXOk1M1S, si XOl1M1S, siXOa0, siXOe0, and siXOf0, as well as comparative siRNAs. CON-siXOf was obtained.

[0504] [Table 4] However, uppercase C, G, U, and A represent the base sequence of a nucleotide, and lowercase m represents the same This indicates that the nucleotide adjacent to the left of the letter 'm' is a methoxy-modified nucleotide. Furthermore, the lowercase letter f is a fluoromodified nucleus, where one nucleotide adjacent to the left of the letter f is fluoromodified. The lowercase letter 's' indicates that there is a thio between the two nucleotides to the left and right of the letter 's'. This indicates that the groups are bonded by phosphate ester groups.

[0505] In the above sequence preparation process, if the target sequence contains unmodified nucleotides, cleavage and Under deprotection conditions, after treatment with aqueous ammonia, the amount of single-stranded nucleic acid was 0.4 ml / μm. After dissolving the product in 1 / 10 N-methylpyrrolidone, add 0.3 ml / μmol triethyl Add the amine and 0.6 ml / μmol of triethylamine hydrofluoride, then add ribose. The 2'-TBDMS protection was removed.

[0506] After the preparation of the above siRNA or siRNA complex is complete, freeze-dry it as a solid powder. Stored in this manner. When using, for example, with sterile water for injection, physiological saline (NS), phosphate buffer (P B) Alternatively, dissolve the solution again in a solution of the desired concentration using a phosphate buffer (PBS), etc., and use. I was able to do it.

[0507] Experimental Example 1 In vitro inhibitory activity of the siRNAs disclosed herein. 10% fetal bovine serum (FBS, Hyclone) and 0.2% by volume of penicillin - Streptomycin (Penicillin-Streptomycin, Gibco) H-DMEM complete medium (Hyclone) supplemented with Invitrogen (H) K293A cells (purchased from Nanjing Kefei Biotechnology Co., Ltd.) were incubated in a 5% CO2 / 95% air-containing solution. The cells were cultured in an incubator at 37°C.

[0508] Kumico Ui-Tei et.al., Functional dissec tion of siRNA sequence by systematic DNA substitution: modified siRNA with a DNA seed arm is a powerful tool for mammali an gene silencing with significantly red uced off-target effect. Nucleic Acids Re In accordance with the method described in search, 2008.36(7), 2136-2151, the test subject Construct rasmids and evaluate each siRNA (siXOa0, siXOe0, siX Of0 and CON-siXOf) were transfected into HEK293A cells, and The expression level of the allulusciferase reporter gene reflects the repressive activity of siRNA. The specific steps are as follows:

[0509] [1] Construction of the test plasmid psiCHECK TM -2(Promega TM ) Using plasmids, the target sequence is s A test plasmid containing an iRNA target sequence was constructed. Regarding the siRNA to be evaluated, the objective was... The arrays are shown below. Target sequence of siXOa0: GAGATGAAGTTCAAGAATA (Sequence ID 731) Target sequence of siXOe0: ACATGGACAACTGCTATAA(Sequence ID 732) The target array of siXOf0: TAGCAAGCTCTCAGTATCA (Sequence ID 733) CON-siXOf target array: CTAGCAAGCTCTCAGTATC (Sequence ID 734)

[0510] Check the target sequence using psiCHECK TM -2 plasmid Xho I / Not I site I did it.

[0511] [2] transfection 8 × 10⁶ HEK293A cells 3 Inoculate cells / well into a 96-well plate and inoculate for 16 hours. Later, when the cell growth density reaches 70-80%, the H-DMEM complete medium in the culture wells is used. Remove all aspirated material, then add 80 μl of Opti-MEM medium (GIBCO) to each well. The culture was then continued for 1.5 hours.

[0512] For each siRNA, the corresponding test plasmid was diluted with DEPC-treated water and incubated for 200°F. g / μl was used as the standard solution for the test plasmid dilution. For each siRNA, siRNA and D The EPC-treated water yielded concentrations (as siRNA) of 10 nM, 3 nM, and 1 nM, respectively. A standard solution was prepared using a certain siRNA dilution.

[0513] For each siRNA, 1 μl of siRNA diluted standard solution with a concentration of 10 nM was used, and the test product was placed in the appropriate container. 0.05 μl of Rasmid diluted standard solution (containing 10 ng of test plasmid) and Opti-M A solution 1A1 containing 10 μl of EM medium was prepared.

[0514] For each siRNA, 1 μl of siRNA diluted standard solution with a concentration of 3 nM was used, and the test plastic was placed in a sample plastic. 0.05 μl of diluted Sumido standard solution (containing 10 ng of test plasmid) and Opti-ME A 1A2 solution containing 10 μl of M medium was prepared.

[0515] For each siRNA, 1 μl of siRNA diluted standard solution with a concentration of 1 nM was used, and the test plastic was placed in a plastic container. 0.05 μl of diluted Sum...

Claims

1. An siRNA that suppresses the expression of the xanthine oxidase (XO) gene in hepatocytes, The siRNA is a siRNA comprising a sense strand and an antisense strand, wherein each nucleotide of the siRNA is independently modified or unmodified. The sense chain and the antisense chain have the same or different lengths. The sense strand has a length of 19 to 23 nucleotides, and the antisense strand has a length of 19 to 26 nucleotides. The sense strand comprises nucleotide sequence I, and the antisense strand comprises nucleotide sequence II, wherein nucleotide sequence I and nucleotide sequence II form a double-stranded region that is substantially or completely inversely complementary, and nucleotide sequence I and nucleotide sequence II are selected from the sequences shown in i) to xi) below. i) The nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 3, and the nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO:

4. 5'-GAGAUGAAGUUCAAGAAUZ 3-3' (Sequence ID 3), 5'-Z 4 AUUCUUGAACUUCAUCUC-3' (Sequence ID 4) However, Z4 is selected from A, U, G, or C, and Z4 is the first nucleotide from the 5' end of the antisense strand, and Z3 is the first nucleotide from the 3' end of nucleotide sequence I. ii) The nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 63, and the nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO:

64. 5'-CAAUAACUGGAAUUUGUAAZ 7-3' (Sequence ID 63), 5'-Z 8 UUACAAAUUCCAGUUAUG-3' (Sequence ID 64) However, Z8 is selected from A, U, G, or C, and Z8 is the first nucleotide from the 5' end of the antisense strand, and Z7 is the first nucleotide from the 3' end of nucleotide sequence I. iii) The nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 123, and the nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO:

124. 5'-CAUUAUCACAAUUGAGAGAZ 11-3' (Sequence ID 123), 5'-Z 12 UCCUCAAUUGUGAUAUUG-3' (Sequence ID 124) However, Z 12 is selected from A, U, G, or C, and Z 12 is the first nucleotide from the 5' end of the antisense strand, and Z 11 is the first nucleotide from the 3' end of nucleotide sequence I. iv) The nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 183, and the nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO:

184. 5'-GGAUCUCUCUCAGAGUAUZ 15-3' (Sequence ID 183), 5'-Z 16 AUACUCUGAGAGAGAUCC-3' (Sequence ID 184) However, Z 16 is selected from A, U, G, or C, and Z 16 is the first nucleotide from the 5' end of the antisense strand, and Z 15 is the first nucleotide from the 3' end of nucleotide sequence I. v) The nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 243, and the nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO:

244. 5'-ACAUGGACACUGCUAUAZ 19-3' (Sequence ID 243), 5'-Z 20 UAUAGCAGUUGUCCAAUGU-3' (Sequence ID 244) However, Z 20 is selected from A, U, G, or C, and Z 20 is the first nucleotide from the 5' end of the antisense strand, and Z 19 is the first nucleotide from the 3' end of nucleotide sequence I. vi) The nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 303, and the nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO:

304. 5'-UAGCAAGCUCUCAGUAUCZ 23-3' (Sequence ID 303), 5'-Z 24 GAUACUGAGAGCUUGCUA-3' (Sequence ID 304) However, Z 24 is selected from A, U, G, or C, and Z 24 is the first nucleotide from the 5' end of the antisense strand, and Z 23 is the first nucleotide from the 3' end of nucleotide sequence I. vii) The nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 363, and the nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO:

364. 5'-AUAAGGUUACUUGUGUUGZ 27-3' (Sequence ID 363), 5'-Z 28 CAACACAAGUAACCUUAU-3' (Sequence ID 364) However, Z28 is selected from A, U, G, or C, and Z28 is the first nucleotide from the 5' end of the antisense strand, and Z27 is the first nucleotide from the 3' end of nucleotide sequence I. viiii) The nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 423, and the nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO:

424. 5'-GAAAAAUCACCUAUGAAGAZ 31-3' (Sequence ID 423), 5'-Z 32 UCUUCAUAGGUGAUUUUC-3' (Sequence ID 424) However, Z 32 is selected from A, U, G, or C, and Z 32 is the first nucleotide from the 5' end of the antisense strand, and Z 31 is the first nucleotide from the 3' end of nucleotide sequence I. ix) The nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 483, and the nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO:

484. 5'-GAUGCUAUAAAGAACACZ 35-3' (Sequence ID 483), 5'-Z 36 GUUGUUCUUAUAGCAUC-3' (Sequence ID 484) However, Z 36 is selected from A, U, G, or C, and Z 36 is the first nucleotide from the 5' end of the antisense strand, and Z 35 is the first nucleotide from the 3' end of nucleotide sequence I. x) The nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 543, and the nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO:

544. 5'-GAACACUCCUUUUAUGGZ 39-3' (Sequence ID 543), 5'-Z 40 CCAUAAAAGGAGUUGUUC-3' (Sequence ID 544) However, Z 40 is selected from A, U, G, or C, and Z 40 is the first nucleotide from the 5' end of the antisense strand, and Z 39 is the first nucleotide from the 3' end of nucleotide sequence I. xi) The nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 603, and the nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO:

604. 5'-CUUGCUCUGAAGUAGAAAAZ 43-3' (Sequence ID 603), 5'-Z 44 UUUCUACUUCAGCAAG-3' (Sequence ID 604) However, Z 44 is selected from A, U, G, or C, and Z 44 is the first nucleotide from the 5' end of the antisense strand, and Z 43 is the first nucleotide from the 3' end of nucleotide sequence I. xi) The nucleotide sequence I is the nucleotide sequence shown in SEQ ID NO: 663, and the nucleotide sequence II is the nucleotide sequence shown in SEQ ID NO:

664. 5'-CUUCUUUGCCCAUCAAAGAZ 47-3' (Sequence ID 663), 5'-Z 48 UCUUUGAUGGCAAAAGAAG-3' (Sequence ID 664) However, Z48 is selected from A, U, G, or C, and Z48 is the first nucleotide from the 5' end of the antisense strand, and Z47 is the first nucleotide from the 3' end of nucleotide sequence I, wherein the siRNA is an siRNA.

2. Z ３ is, Z ４ It is a complementary nucleotide, Or, Z ７ is, Z ８ It is a complementary nucleotide, Or, Z １１ is, Z １２ It is a complementary nucleotide, Or, Z １５ is, Z １６ It is a complementary nucleotide, Alternatively, Z １９ is Z ２０ and is a nucleotide complementary to Z, Or, Z ２３ is, Z ２４ It is a complementary nucleotide, Or, Z ２７ is, Z ２８ It is a complementary nucleotide, Or, Z ３１ is, Z ３２ It is a complementary nucleotide, Or, Z ３５ is, Z ３６ It is a complementary nucleotide, Or, Z ３９ is, Z ４０ It is a complementary nucleotide, Or, Z ４３ is, Z ４４ It is a complementary nucleotide, Or, Z ４７ is, Z ４８ The siRNA according to claim 1, which is a nucleotide complementary to the siRNA.

3. The siRNA according to claim 1, wherein the sense strand further comprises nucleotide sequence III, and the antisense strand further comprises nucleotide sequence IV, the lengths of nucleotide sequence III and nucleotide sequence IV are independently 1 to 4 nucleotides, nucleotide sequence III is attached to the 5' end of nucleotide sequence I, and nucleotide sequence IV is attached to the 3' end of nucleotide sequence II, and nucleotide sequence III and nucleotide sequence IV are of equal length and substantially inversely complementary or completely inversely complementary, wherein substantially inversely complementary means that there is one or fewer base mismatches between the two nucleotide sequences, and completely inversely complementary means that there are no mismatches between the two nucleotide sequences.

4. The nucleotide sequence I is the nucleotide sequence shown in Sequence ID No. 3, and the nucleotide sequences III and IV are both 1 nucleotide in length and the base of nucleotide sequence III is U, or the nucleotide sequences III and IV are both 2 nucleotides in length and the base sequence of nucleotide sequence III is UU from the 5' end to the 3' end, or the nucleotide sequences III and IV are both 3 nucleotides in length and the base sequence of nucleotide sequence III is AUU from the 5' end to the 3' end, or the nucleotide sequences III and IV are both 4 nucleotides in length and the base sequence of nucleotide sequence III is CAUU from the 5' end to the 3' end. Alternatively, nucleotide sequence I is the nucleotide sequence shown in Sequence ID No. 63, and nucleotide sequences III and IV are both 1 nucleotide in length, with the base of nucleotide sequence III being A, or nucleotide sequences III and IV are both 2 nucleotides in length, with the base sequence of nucleotide sequence III being AA from the 5' end to the 3' end, or nucleotide sequences III and IV are both 3 nucleotides in length, with the base sequence of nucleotide sequence III being UAA from the 5' end to the 3' end, or nucleotide sequences III and IV are both 4 nucleotides in length, with the base sequence of nucleotide sequence III being GUAA from the 5' end to the 3' end. Alternatively, nucleotide sequence I is the nucleotide sequence shown in Sequence ID No. 123, and nucleotide sequences III and IV are both 1 nucleotide in length, with the base of nucleotide sequence III being C, or nucleotide sequences III and IV are both 2 nucleotides in length, with the base sequence of nucleotide sequence III being GC from the 5' end to the 3' end, or nucleotide sequences III and IV are both 3 nucleotides in length, with the base sequence of nucleotide sequence III being AGC from the 5' end to the 3' end, or nucleotide sequences III and IV are both 4 nucleotides in length, with the base sequence of nucleotide sequence III being CAGC from the 5' end to the 3' end. Alternatively, nucleotide sequence I is the nucleotide sequence shown in Sequence ID No. 183, and nucleotide sequences III and IV are both 1 nucleotide in length, with the base of nucleotide sequence III being A, or nucleotide sequences III and IV are both 2 nucleotides in length, with the base sequence of nucleotide sequence III being CA from the 5' end to the 3' end, or nucleotide sequences III and IV are both 3 nucleotides in length, with the base sequence of nucleotide sequence III being CCA from the 5' end to the 3' end, or nucleotide sequences III and IV are both 4 nucleotides in length, with the base sequence of nucleotide sequence III being CCCA from the 5' end to the 3' end. Alternatively, nucleotide sequence I is the nucleotide sequence shown in Sequence ID No. 243, and nucleotide sequences III and IV are both 1 nucleotide in length, with the base of nucleotide sequence III being C, or nucleotide sequences III and IV are both 2 nucleotides in length, with the base sequence of nucleotide sequence III being CC from the 5' end to the 3' end, or nucleotide sequences III and IV are both 3 nucleotides in length, with the base sequence of nucleotide sequence III being UCC from the 5' end to the 3' end, or nucleotide sequences III and IV are both 4 nucleotides in length, with the base sequence of nucleotide sequence III being UUCC from the 5' end to the 3' end. Alternatively, nucleotide sequence I is the nucleotide sequence shown in Sequence ID No. 303, and nucleotide sequences III and IV are both 1 nucleotide in length, with the base of nucleotide sequence III being C, or nucleotide sequences III and IV are both 2 nucleotides in length, with the base sequence of nucleotide sequence III being CC from the 5' end to the 3' end, or nucleotide sequences III and IV are both 3 nucleotides in length, with the base sequence of nucleotide sequence III being GCC from the 5' end to the 3' end, or nucleotide sequences III and IV are both 4 nucleotides in length, with the base sequence of nucleotide sequence III being UGCC from the 5' end to the 3' end. Alternatively, nucleotide sequence I is the nucleotide sequence shown in Sequence ID No. 363, and nucleotide sequences III and IV are both 1 nucleotide in length, with the base of nucleotide sequence III being G, or nucleotide sequences III and IV are both 2 nucleotides in length, with the base sequence of nucleotide sequence III being GG from the 5' end to the 3' end, or nucleotide sequences III and IV are both 3 nucleotides in length, with the base sequence of nucleotide sequence III being AGG from the 5' end to the 3' end, or nucleotide sequences III and IV are both 4 nucleotides in length, with the base sequence of nucleotide sequence III being AAGG from the 5' end to the 3' end. Alternatively, nucleotide sequence I is the nucleotide sequence shown in Sequence ID No. 423, and nucleotide sequences III and IV are both 1 nucleotide in length, with the base of nucleotide sequence III being U, or nucleotide sequences III and IV are both 2 nucleotides in length, with the base sequence of nucleotide sequence III being GU from the 5' end to the 3' end, or nucleotide sequences III and IV are both 3 nucleotides in length, with the base sequence of nucleotide sequence III being GGU from the 5' end to the 3' end, or nucleotide sequences III and IV are both 4 nucleotides in length, with the base sequence of nucleotide sequence III being GGGU from the 5' end to the 3' end. Alternatively, nucleotide sequence I is the nucleotide sequence shown in Sequence ID No. 483, and nucleotide sequences III and IV are both 1 nucleotide in length, with the base of nucleotide sequence III being G, or nucleotide sequences III and IV are both 2 nucleotides in length, with the base sequence of nucleotide sequence III being AG from the 5' end to the 3' end, or nucleotide sequences III and IV are both 3 nucleotides in length, with the base sequence of nucleotide sequence III being GAG from the 5' end to the 3' end, or nucleotide sequences III and IV are both 4 nucleotides in length, with the base sequence of nucleotide sequence III being UGAG from the 5' end to the 3' end. Alternatively, nucleotide sequence I is the nucleotide sequence shown in Sequence ID No. 543, and nucleotide sequences III and IV are both 1 nucleotide in length, with the base of nucleotide sequence III being A, or nucleotide sequences III and IV are both 2 nucleotides in length, with the base sequence of nucleotide sequence III being AA from the 5' end to the 3' end, or nucleotide sequences III and IV are both 3 nucleotides in length, with the base sequence of nucleotide sequence III being AAA from the 5' end to the 3' end, or nucleotide sequences III and IV are both 4 nucleotides in length, with the base sequence of nucleotide sequence III being UAAA from the 5' end to the 3' end. Alternatively, nucleotide sequence I is the nucleotide sequence shown in Sequence ID No. 603, and nucleotide sequences III and IV are both 1 nucleotide in length, with the base of nucleotide sequence III being G, or nucleotide sequences III and IV are both 2 nucleotides in length, with the base sequence of nucleotide sequence III being GG from the 5' end to the 3' end, or nucleotide sequences III and IV are both 3 nucleotides in length, with the base sequence of nucleotide sequence III being UGG from the 5' end to the 3' end, or nucleotide sequences III and IV are both 4 nucleotides in length, with the base sequence of nucleotide sequence III being GUGG from the 5' end to the 3' end. Alternatively, the siRNA according to claim 3, wherein nucleotide sequence I is the nucleotide sequence shown in Sequence ID No. 663, nucleotide sequences III and IV are both 1 nucleotide in length and the base of nucleotide sequence III is U, or nucleotide sequences III and IV are both 2 nucleotides in length and the base sequence of nucleotide sequence III is AU from the 5' end to the 3' end, or nucleotide sequences III and IV are both 3 nucleotides in length and the base sequence of nucleotide sequence III is UAU from the 5' end to the 3' end, or nucleotide sequences III and IV are both 4 nucleotides in length and the base sequence of nucleotide sequence III is CUAU from the 5' end to the 3' end.

5. The siRNA according to claim 1, wherein the antisense strand further comprises a nucleotide sequence V, the nucleotide sequence V having a length of 1 to 3 nucleotides, and is bound to the 3' end of the antisense strand, constituting the 3' protruding end of the antisense strand.

6. The sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 5, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

6. 5'-GAGAUGAAGUUCAAGAAUZ ３ -3' (Sequence ID 5), 5'-Z ４ AUUCUUGAACUUCAUCUCAA-3' (Sequence ID 6) Alternatively, the sense strand of the siRNA includes the nucleotide sequence shown in SEQ ID NO: 7, and the antisense strand includes the nucleotide sequence shown in SEQ ID NO:

8. 5'-UUGAGAUGAAGUUCAAGAAUZ ３ -3' (Sequence ID 7), 5'-Z ４ AUUCUUGAACUUCAUCUCAAUG-3' (Sequence ID 8) However, the above Z ４ Z is the first nucleotide at the 5' end of the antisense strand. ４ is selected from A, U, G, or C, and Z ３ is, Z ４ It is a complementary nucleotide, Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 65, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

66. 5'-CAUAACUGGAAUUUGUAAZ ７ -3' (Sequence ID 65), 5'-Z ８ UUACAAAAUUCCAGUUAUGUU-3' (Sequence ID 66) Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 67, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

68. 5'-AACAUAACUGGAAUUUGUAAZ ７ -3' (Sequence ID 67), 5'-Z ８ UUACAAAAUUCCAGUUAUGUUAC-3' (Sequence ID 68) However, the above Z ８ Z is the first nucleotide at the 5' end of the antisense strand. ８ is selected from A, U, G, or C, and Z ７ is, Z ８ It is a complementary nucleotide, Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 125, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

126. 5'-CAUUAUCACAAUUGAGAZ １１ -3' (Sequence ID 125), 5'-Z １２ UCCUCAAUUGUGAUAAUGGC-3' (Sequence ID 126) Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 127, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

128. 5'-GCCAUUAUCACAAUUGAGGAZ １１ -3' (Sequence ID 127), 5'-Z １２ UCCUCAAUUGUGAUAAUGGCUG-3' (Sequence ID 128) However, the above Z １２ Z is the first nucleotide at the 5' end of the antisense strand. １２ is selected from A, U, G, or C, and Z １１ is, Z １２ It is a complementary nucleotide, Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 185, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

186. 5'-GGAUCUCUCUCAGAGUAUZ １５ -3' (Sequence ID 185), 5'-Z １６ AUACUCUGAGAGAGAGAUCCUG-3' (Sequence ID 186) Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 187, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

188. 5'-CAGGAUCUCUCUCAGAGUAUZ １５ -3' (Sequence ID 187), 5'-Z １６ AUACUCUGAGAGAGAUCCUGGG-3' (Sequence ID 188) However, the above Z １６ Z is the first nucleotide at the 5' end of the antisense strand. １６ is selected from A, U, G, or C, and Z １５ is, Z １６ It is a complementary nucleotide, Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 245, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

246. 5'-ACAUGGACAACUGCUAUAZ １９ -3' (Sequence ID 245), 5'-Z ２０ UAUAGCAGUUGUCCAAUGUGG-3' (Sequence ID 246) Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 247, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

248. 5'-CCACAUGGACAACUGCUAUAZ １９ -3' (Sequence ID 247), 5'-Z ２０ UAUAGCAGUUGUCCAAUGUGGAA-3' (Sequence ID 248) However, the above Z ２０ Z is the first nucleotide at the 5' end of the antisense strand. ２０ is selected from A, U, G, or C, and Z １９ is, Z ２０ It is a complementary nucleotide, Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 305, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

306. 5'-UAGCAAGCUCUCAGUAUCZ ２３ -3' (Sequence ID 305), 5'-Z ２４ GAUACUGAGAGCUUGCUAGG-3' (Sequence ID 306) Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 307, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

308. 5'-CCUAGCAAGCUCUCAGUAUCZ ２３ -3' (Sequence ID 307), 5'-Z ２４ GAUACUGAGAGCUUGCUAGCA-3' (Sequence ID 308) However, the above Z ２４ Z is the first nucleotide at the 5' end of the antisense strand. ２４ is selected from A, U, G, or C, and Z ２３ is, Z ２４ It is a complementary nucleotide, Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 365, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

366. 5'-AUAAGGUUACUUGUGUUGZ ２７ -3' (Sequence ID 365), 5'-Z ２８ CAACACAAGUAACCUUAUCC-3' (Sequence ID 366) Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 367, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

368. 5'-GGAUAAGGUUACUUGUGUUGZ ２７ -3' (Sequence ID 367), 5'-Z ２８ CAACACAAGUAACCUUAUCCUU-3' (Sequence ID 368) However, the above Z ２８ Z is the first nucleotide at the 5' end of the antisense strand. ２８ is selected from A, U, G, or C, and Z ２７ is, Z ２８ It is a complementary nucleotide, Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 425, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

426. 5'-GAAAAUCACCUAUGAAGAZ ３１ -3' (Sequence ID 425), 5'-Z ３２ UCUUCAUAGGUGAUUUUCAC-3' (Sequence ID 426) Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 427, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

428. 5'-GUGAAAAUCACCUAUGAAGAZ ３１ -3' (Sequence ID 427), 5'-Z ３２ UCUUCAUAGGUGAAUUUUCACCC-3' (Sequence ID 428) However, the above Z ３２ Z is the first nucleotide at the 5' end of the antisense strand. ３２ is selected from A, U, G, or C, and Z ３１ is, Z ３２ It is a complementary nucleotide, Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 485, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

486. 5'-GAUGCUAUAAAGAACAACZ ３５ -3' (Sequence ID 485), 5'-Z ３６ GUUGUUCUUAUAGCAUCCU-3' (SEQ ID NO: 486) Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 487, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

488. 5'-AGGAUGCUAUAAAGAACAACZ ３５ -3' (Sequence ID 487), 5'-Z ３６ GUUGUUCUUAUAGCAUCCUCA-3' (Sequence ID 488) However, the above Z ３６ Z is the first nucleotide at the 5' end of the antisense strand. ３６ is selected from A, U, G, or C, and Z ３５ is, Z ３６ It is a complementary nucleotide, Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 545, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

546. 5'-GAA CAA CUC CUU UUA UGG Z ３９ -3' (SEQ ID NO: 545), 5'-Z ４０ CCAUAAAAGGAGUUGUUCU-3' (Sequence ID 546) Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 547, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

548. 5'-AAGAACAACUCCUUUUAUGGZ ３９ -3' (Sequence ID 547), 5'-Z ４０ CCAUAAAAGGAGUUGUUCUUA-3' (Sequence ID 548) However, the above Z ４０ Z is the first nucleotide at the 5' end of the antisense strand. ４０ is selected from A, U, G, or C, and Z ３９ is, Z ４０ It is a complementary nucleotide, Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 605, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

606. 5'-CUUGCUCUGAAGUAGAAAZ ４３ -3' (Sequence ID 605), 5'-Z ４４ UUUCUACUUCAGAGCAAGCC-3' (Sequence ID 606) Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 607, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

608. 5'-GGCUUGCUCUGAAGUAGAAAZ ４３ -3' (Sequence ID 607), 5'-Z ４４ UUUCUACUUCAGAGCAAGCCAC-3' (Sequence ID 608) However, the above Z ４４ Z is the first nucleotide at the 5' end of the antisense strand. ４４ is selected from A, U, G, or C, and Z ４３ is, Z ４４ It is a complementary nucleotide, Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 665, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

666. 5'-CUUCUUUGCCAUCAAAGAZ ４７ -3' (Sequence ID 665), 5'-Z ４８ UCUUUGAUGGCAAAAGAAGAU-3' (Sequence ID 666) Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 667, and the antisense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO:

668. 5'-AUCUUCUUUGCCAUCAAAGAZ ４７ -3' (Sequence ID 667), 5'-Z ４８ UCUUUGAUGGCAAAAGAAGAUAG-3' (Sequence ID 668) However, the above Z ４８ Z is the first nucleotide at the 5' end of the antisense strand. ４８ is selected from A, U, G, or C, and Z ４７ is, Z ４８ The siRNA according to claim 5, which is a nucleotide complementary to the siRNA.

7. Each nucleotide in the sense strand and the antisense strand is independently either a fluoromodified nucleotide or an unfluoromodified nucleotide. The siRNA according to claim 1, wherein the fluoromodified nucleotides are located in nucleotide sequence I and nucleotide sequence II, and from the 5' end toward the 3' end, at least the nucleotides at positions 7, 8, and 9 of nucleotide sequence I are fluoromodified nucleotides, and from the 5' end toward the 3' end, at least the nucleotides at positions 2, 6, 14, and 16 of nucleotide sequence II are fluoromodified nucleotides.

8. Each non-fluoromodified nucleotide is independently selected from nucleotides or nucleotide analogs in which the hydroxyl group at the 2' position of the ribose group of the nucleotide is substituted with a non-fluorinated group, or A nucleotide in which the hydroxyl group at the 2' position of the ribose group of the nucleotide is substituted with a nonfluorine group is one selected from 2'-alkoxy-modified nucleotides, 2'-substituted alkoxy-modified nucleotides, 2'-alkyl-modified nucleotides, 2'-substituted alkyl-modified nucleotides, 2'-amino-modified nucleotides, 2'-substituted amino-modified nucleotides, and 2'-deoxynucleotides, and the nucleotide analog is one selected from isonucleotides, LNA, ENA, cET, UNA, and GNA, or The siRNA according to claim 7, wherein each non-fluoromodified nucleotide is a methoxy-modified nucleotide, and the methoxy-modified nucleotide refers to a nucleotide in which the 2'-hydroxyl group of the ribose group is replaced with methoxy.

9. From the 5' end toward the 3' end, the nucleotides at positions 5, 7, 8 and 9 of nucleotide sequence I in the sense strand of the siRNA are fluoromodified nucleotides, and the nucleotides at the remaining positions of the siRNA sense strand are methoxymodified nucleotides; From the 5' end toward the 3' end, the nucleotides at positions 2, 6, 8, 9, 14 and 16 of nucleotide sequence II in the antisense strand of the siRNA are fluoromodified nucleotides, and the nucleotides at the remaining positions of the siRNA antisense strand are methoxymodified nucleotides. Alternatively, from the 5' end to the 3' end, the nucleotides at positions 5, 7, 8 and 9 of nucleotide sequence I in the sense strand of the siRNA are fluoromodified nucleotides, and the remaining nucleotides in the sense strand of the siRNA are methoxymodified nucleotides; and from the 5' end to the 3' end, the nucleotides at positions 2, 6, 14 and 16 of nucleotide sequence II in the antisense strand of the siRNA are fluoromodified nucleotides, and the remaining nucleotides in the antisense strand of the siRNA are methoxymodified nucleotides. Alternatively, the siRNA according to claim 8, wherein, from the 5' end to the 3' end, the nucleotides at positions 7, 8 and 9 of nucleotide sequence I in the sense strand of the siRNA are fluoromodified nucleotides, and the nucleotides at the remaining positions of the sense strand of the siRNA are methoxymodified nucleotides, and from the 5' end to the 3' end, the nucleotides at positions 2, 6, 14 and 16 of nucleotide sequence II in the antisense strand of the siRNA are fluoromodified nucleotides, and the nucleotides at the remaining positions of the antisense strand of the siRNA are methoxymodified nucleotides.

10. In the siRNA, at least one phosphate ester group is a thiophosphate ester group, and the thiophosphate ester group is Between the first and second nucleotides from the 5' end of the sense strand, Between the second and third nucleotides from the 5' end of the sense strand, Between the first and second nucleotides from the 3' end of the sense strand, Between the second and third nucleotides from the 3' end of the sense strand, Between the first and second nucleotides from the 5' end of the aforementioned antisense strand, Between the second and third nucleotides from the 5' end of the aforementioned antisense strand, Between the first and second nucleotides from the 3' end of the antisense strand, and The siRNA according to claim 1, wherein the siRNA is bound to at least one of the group consisting of the space between the second and third nucleotides from the 3' end of the antisense strand.

11. The 5' terminal nucleotide of the antisense strand of the siRNA is a 5'-phosphate nucleotide or a 5'-phosphate analog modified nucleotide. Alternatively, the 5'-phosphate nucleotide is a nucleotide having the structure shown in formula (2), and the 5'-phosphate analog modified nucleotide is selected from nucleotides having the structure shown in any one of formulas (3) to (6). 【Chemistry 1】 The siRNA according to claim 1, wherein R is selected from H, OH, a methoxy group or fluorine, and Base represents a base and is selected from A, U, C, G or T.

12. The sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 9, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

10. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 11, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

12. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 69, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

70. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 71, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

72. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 129, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

130. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 131, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

132. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 189, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

190. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 191, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

192. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 249, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

250. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 251, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

252. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 309, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

310. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 311, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

312. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 369, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

370. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 371, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

372. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 429, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

430. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 431, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

432. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 489, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

490. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 491, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

492. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 549, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

550. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 551, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

552. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 609, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

610. Alternatively, the sense strand of the siRNA may contain the nucleotide sequence shown in SEQ ID NO: 611, and the antisense strand may contain the nucleotide sequence shown in SEQ ID NO: 612, or The sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 13, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

14. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 15, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

16. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 17, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

18. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 19, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

20. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 21, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

22. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 23, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

24. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 73, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

74. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 75, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

76. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 77, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

78. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 79, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

80. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 81, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

82. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 83, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

84. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 133, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

134. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 135, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

136. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 137, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

138. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 139, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

140. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 141, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

142. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 143, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

144. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 193, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

194. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 195, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

196. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 197, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

198. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 199, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

200. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 201, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

202. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 203, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

204. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 253, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

254. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 255, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

256. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 257, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

258. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 259, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

260. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 261, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

262. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 263, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

264. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 313, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

314. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 315, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

316. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 317, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

318. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 319, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

320. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 321, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

322. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 323, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

324. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 373, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

374. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 375, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

376. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 377, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

378. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 379, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

380. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 381, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

382. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 383, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

384. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 433, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

434. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 435, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

436. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 437, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

438. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 439, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

440. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 441, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

442. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 443, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

444. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 493, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

494. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 495, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

496. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 497, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

498. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 499, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

500. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 501, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

502. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 503, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

504. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 553, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

554. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 555, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

556. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 557, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

558. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 559, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

560. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 561, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

562. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 563, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

564. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 613, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

614. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 615, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

616. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 617, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

618. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 619, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

620. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 621, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

622. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 623, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

624. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 673, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

674. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 675, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

676. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 677, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

678. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 679, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

680. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 681, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

682. Alternatively, the sense strand of the siRNA may contain the nucleotide sequence shown in SEQ ID NO: 683, and the antisense strand may contain the nucleotide sequence shown in SEQ ID NO: 684, or The sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 25, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

26. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 27, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

28. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 29, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

30. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 31, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

32. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 33, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

34. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 35, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

36. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 85, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

86. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 87, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

88. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 89, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

90. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 91, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

92. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 93, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

94. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 95, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

96. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 145, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

146. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 147, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

148. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 149, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

150. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 151, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

152. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 153, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

154. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 155, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

156. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 205, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

206. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 207, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

208. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 209, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

210. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 211, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

212. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 213, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

214. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 215, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

216. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 265, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

266. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 267, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

268. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 269, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

270. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 271, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

272. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 273, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

274. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 275, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

276. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 325, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

326. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 327, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

328. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 329, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

330. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 331, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

332. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 333, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

334. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 335, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

336. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 385, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

386. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 387, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

388. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 389, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

390. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 391, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

392. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 393, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

394. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 395, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

396. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 445, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

446. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 447, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

448. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 449, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

450. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 451, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

452. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 453, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

454. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 455, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

456. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 505, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

506. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 507, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

508. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 509, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

510. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 511, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

512. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 513, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

514. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 515, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

516. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 565, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

566. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 567, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

568. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 569, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

570. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 571, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

572. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 573, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

574. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 575, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

576. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 625, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

626. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 627, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

628. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 629, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

630. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 631, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

632. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 633, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

634. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 635, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

636. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 685, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

686. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 687, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

688. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 689, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

690. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 691, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

692. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 693, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

694. Alternatively, the sense strand of the siRNA may contain the nucleotide sequence shown in SEQ ID NO: 695, and the antisense strand may contain the nucleotide sequence shown in SEQ ID NO: 696, or The sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 37, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

38. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 39, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

40. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 41, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

42. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 43, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

44. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 45, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

46. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 47, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

48. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 49, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

50. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 51, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

52. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 53, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

54. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 55, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

56. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 57, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

58. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 59, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

60. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 97, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

98. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 99, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

100. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 101, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

102. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 103, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

104. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 105, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

106. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 107, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

108. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 109, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

110. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 111, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

112. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 113, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

114. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 115, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

116. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 117, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

118. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 119, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

120. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 157, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

158. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 159, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

160. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 161, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

162. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 163, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

164. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 165, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

166. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 167, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

168. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 169, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

170. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 171, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

172. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 173, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

174. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 175, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

176. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 177, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

178. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 179, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

180. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 217, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

218. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 219, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

220. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 221, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

222. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 223, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

224. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 225, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

226. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 227, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

228. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 229, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

230. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 231, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

232. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 233, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

234. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 235, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

236. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 237, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

238. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 239, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

240. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 277, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

278. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 279, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

280. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 281, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

282. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 283, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

284. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 285, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

286. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 287, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

288. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 289, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

290. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 291, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

292. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 293, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

294. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 295, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

296. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 297, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

298. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 299, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

300. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 337, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

338. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 339, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

340. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 341, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

342. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 343, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

344. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 345, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

346. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 347, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

348. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 349, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

350. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 351, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

352. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 353, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

354. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 355, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

356. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 357, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

358. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 359, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

360. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 397, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

398. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 399, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

400. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 401, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

402. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 403, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

404. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 405, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

406. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 407, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

408. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 409, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

410. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 411, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

412. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 413, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

414. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 415, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

416. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 417, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

418. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 419, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

420. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 457, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

458. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 459, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

460. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 461, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

462. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 463, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

464. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 465, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

466. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 467, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

468. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 469, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

470. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 471, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

472. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 473, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

474. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 475, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

476. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 477, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

478. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 479, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

480. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 517, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

518. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 519, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

520. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 521, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

522. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 523, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

524. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 525, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

526. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 527, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

528. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 529, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

530. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 531, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

532. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 533, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

534. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 535, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

536. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 537, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

538. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 539, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

540. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 577, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

578. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 579, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

580. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 581, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

582. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 583, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

584. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 585, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

586. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 587, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

588. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 589, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

590. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 591, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

592. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 593, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

594. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 595, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

596. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 597, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

598. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 599, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

600. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 637, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

638. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 639, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

640. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 641, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

642. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 643, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

644. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 645, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

646. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 647, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

648. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 649, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

650. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 651, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

652. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 653, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

654. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 655, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

656. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 657, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

658. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 659, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

660. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 697, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

698. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 699, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

700. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 701, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

702. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 703, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

704. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 705, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

706. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 707, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

708. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 709, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

710. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 711, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

712. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 713, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

714. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 715, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

716. Alternatively, the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 717, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

718. Alternatively, the siRNA according to claim 1, wherein the sense strand of the siRNA comprises the nucleotide sequence shown in SEQ ID NO: 719, and the antisense strand comprises the nucleotide sequence shown in SEQ ID NO:

720.

13. The siRNA described in claim 1 and a pharmaceutically acceptable carrier are included. A drug composition in which the weight ratio of the siRNA to a pharmaceutically acceptable carrier is 1:(1 to 500).

14. A siRNA complex comprising the siRNA described in claim 1 and a complex group that is bound to the siRNA.

15. A composition for the treatment and / or prevention of uric acid metabolic disorders or diseases or physiological conditions resulting therefrom, comprising the siRNA described in any one of claims 1 to 12, the drug composition described in claim 13, and / or the siRNA complex described in claim 14.

16. The composition according to claim 15, wherein the disease or physiological condition due to the uric acid metabolic disorder is hyperuricemia or gout.

17. A composition for suppressing the expression of the XO gene in hepatocytes, comprising the siRNA described in any one of claims 1 to 12, the drug composition described in claim 13, and / or the siRNA complex described in claim 14.

Images