Double-stranded oligonucleotides, conjugates, pharmaceutical compositions and their uses that inhibit URAT1 gene expression

Abstract

This invention relates to the field of RNAi technology, specifically providing a double-stranded oligonucleotide, conjugate, pharmaceutical composition, and their uses for inhibiting URAT1 gene expression. The provided double-stranded oligonucleotide, conjugate, and pharmaceutical composition can inhibit URAT1 gene expression, reduce URAT1 protein levels, and significantly lower blood uric acid, thus aiding in the treatment and / or prevention of pathological conditions or diseases associated with URAT1 expression, including hyperuricemia or gout.

Description

Double-stranded oligonucleotide conjugate for inhibiting expression of URAT1 gene, pharmaceutical composition and use thereof

[0001] Cross-reference to related applications

[0002] The present disclosure claims priority to the Chinese patent application No. 202410537237.7, filed on April 30, 2024, entitled “Oligonucleotide conjugate for inhibiting expression of URAT1 and use thereof”, and the Chinese patent application No. 202510436069.7, filed on April 8, 2025, entitled “Double-stranded oligonucleotide conjugate for inhibiting expression of URAT1 gene, pharmaceutical composition and use thereof”, the contents of which are incorporated herein by reference in their entirety. TECHNICAL FIELD

[0003] The present disclosure belongs to the field of RNAi technology, and specifically provides a double-stranded oligonucleotide conjugate for inhibiting expression of URAT1 gene, pharmaceutical composition and use thereof. BACKGROUND

[0004] Urate Transporter 1 (URAT1) is a major transporter that controls the reabsorption of urate in the kidney and is a key target for the pathogenesis of hyperuricemia. Existing drugs for treating hyperuricemia by inhibiting URAT1 usually reduce the reabsorption to reduce blood uric acid levels to treat hyperuricemia and gout. However, such inhibitors usually have strong hepatotoxicity and kidney damage.

[0005] RNAi is one of the more mature treatment methods. Reducing the expression of the gene encoding URAT1 protein can reduce urate reabsorption, promote increased urate excretion, and thus reduce blood uric acid. However, there is no RNAi drug targeting URAT1 on the market. SUMMARY

[0006] In view of this, the present disclosure provides a double-stranded oligonucleotide conjugate for inhibiting expression of URAT1, a pharmaceutical composition and use thereof.

[0007] In a first aspect of the present disclosure, a double-stranded oligonucleotide for inhibiting expression of URAT1 gene is provided, the double-stranded oligonucleotide comprising a sense strand and an antisense strand, the length of the sense strand and the antisense strand being 17-30 nucleotides, respectively, and the sense strand and the antisense strand being at least partially complementary; wherein, starting from the 5' end, the antisense strand comprises at least 17 consecutive nucleotides differing from no more than 3 nucleotides in the 1st-19th nucleotides of any one of the nucleotide sequences shown in SEQ ID NO. 70-138, 287-296.

[0008] In a second aspect of the present disclosure, the present disclosure provides a conjugate comprising the double-stranded oligonucleotide according to the first aspect of the present disclosure and one or more targeting ligand groups conjugated to the double-stranded oligonucleotide.

[0009] In a third aspect of the present disclosure, the present disclosure provides a pharmaceutical composition comprising the double-stranded oligonucleotide according to the first aspect of the present disclosure, and / or the conjugate according to the second aspect of the present disclosure.

[0010] In a fourth aspect of the present disclosure, the present disclosure provides use of the double-stranded oligonucleotide according to the first aspect of the present disclosure, and / or the conjugate according to the second aspect of the present disclosure, and / or the pharmaceutical composition according to the third aspect of the present disclosure in the manufacture of a medicament for inhibiting the expression of URAT1 gene.

[0011] In a fifth aspect of the present disclosure, the present disclosure provides use of the double-stranded oligonucleotide according to the first aspect of the present disclosure, and / or the conjugate according to the second aspect of the present disclosure, and / or the pharmaceutical composition according to the third aspect of the present disclosure in the manufacture of a medicament for treating and / or preventing a pathological condition or disease associated with the expression of URAT1.

[0012] In a sixth aspect of the present disclosure, the present disclosure provides a method for inhibiting the expression of URAT1 gene, comprising administering to a subject an effective amount of the double-stranded oligonucleotide according to the first aspect of the present disclosure, and / or the conjugate according to the second aspect of the present disclosure, and / or the pharmaceutical composition according to the third aspect of the present disclosure.

[0013] In a seventh aspect of the present disclosure, the present disclosure provides a method for preventing and / or treating hyperuricemia or gout, comprising administering to a subject an effective amount of the double-stranded oligonucleotide according to the first aspect of the present disclosure, and / or the conjugate according to the second aspect of the present disclosure, and / or the pharmaceutical composition according to the third aspect of the present disclosure.

[0014] The double-stranded oligonucleotide, the conjugate and the pharmaceutical composition provided by the present disclosure can inhibit the expression of URAT1 gene, reduce the content of URAT1 protein, and significantly reduce blood uric acid, which is helpful for treating and / or preventing a pathological condition or disease associated with the expression of URAT1, including hyperuricemia or gout. DETAILED DESCRIPTION

[0015] The technical solutions in the embodiments of the present disclosure will be described clearly and completely below, and those skilled in the art can implement the process parameters by referring to the content herein and making appropriate improvements.

[0016] General definitions and terms:

[0017] Unless otherwise defined, all technical and scientific terms used in the present disclosure have the same meaning as commonly understood by one of ordinary skill in the art. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below. The publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0018] In the present disclosure, the terms "comprising" or "including," are open-ended, in the present disclosure are used to mean "including but not limited to," and are synonymous with the phrase "consisting essentially of," as long as the phrase is not explicitly present. Thus, use of "including" or "comprising" indicates that various alternatives, modifications and equivalents can be used in the present disclosure.

[0019] In the present disclosure, the terms "optionally," "optional," or "optional" generally mean that the subsequently described event or circumstance can or can not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0020] In the present disclosure, the term "small interfering RNA (siRNA)" is a double-stranded RNA of 17 to 25 nucleotides in length, comprising a sense strand and an antisense strand. siRNA mediates the targeted cleavage of RNA transcripts of the RISC pathway by forming a silencing complex (RNA-induced silencing complex, RISC). Specifically, siRNA directs the specific degradation of mRNA sequences through a known RNA interference (RNAi) process, inhibiting the translation of mRNA into amino acids and conversion into protein. For example, siRNA can modulate (e.g., inhibit) the expression of URAT1 in a cell.

[0021] In the present disclosure, the terms "sequence" and "nucleotide sequence" refer to a series of nucleobases or nucleotides. As used in the present disclosure, a "base," "nucleotide base," or "nucleobase" is a pyrimidine or purine compound that is a component of a nucleotide and includes the purine bases adenine and guanine, and the pyrimidine bases cytosine, thymine, and uracil. Nucleobases can be further modified. The synthesis of modified nucleobases, including phosphoramidite compounds of modified nucleobases, is known in the art.

[0022] In the present disclosure, the term "double-stranded oligonucleotide" refers to a double-stranded structure formed by partial or complete base-pairing of two oligonucleotides, including a sense strand and an antisense strand, which can or can not be of the same length, as long as there is at least a region of partial base-pairing to form a duplex region. An oligonucleotide having a double-stranded structure is within the scope of the double-stranded oligonucleotide of the present disclosure. In the present disclosure, the nucleotides in a double-stranded oligonucleotide can be modified or unmodified, and when referring to modified nucleotides, the modifications are not intended to be specific to the position of the modification unless otherwise specified. In the present disclosure, a double-stranded oligonucleotide can further comprise modified linkages between the nucleotides, and a double-stranded oligonucleotide comprising modified linkages between the nucleotides is within the scope of the double-stranded oligonucleotide of the present disclosure. In the present disclosure, a double-stranded oligonucleotide can further comprise a molecule or a moiety acceptable in the art to improve the properties of the double-stranded oligonucleotide, such as a conjugate-forming ligand.

[0023] In the present disclosure, the term "antisense strand (or guide strand)" includes a region that is substantially complementary to a target sequence. The term "sense strand (or passenger strand)" refers to an iRNA strand that contains a sequence that is substantially complementary to the antisense strand. The term "substantially complementary" refers to complete complementarity or at least partial complementarity, for example, the antisense strand is completely complementary or at least partially complementary to the target sequence. In the case of partial complementarity, mismatches can exist within the interior of the molecule or in the terminal regions, and the most tolerated mismatches are in the terminal regions, for example, within 5, 4, 3, or 2 nucleotides of the 5'- and / or 3' end of the iRNA. It is noted that "at least partially substantially complementary" to an mRNA by an antisense strand means that the antisense strand has a polynucleotide that is substantially complementary to a contiguous portion of the mRNA of interest.

[0024] In the present disclosure, the term "target sequence" refers to a contiguous portion of the nucleotide sequence of an mRNA molecule formed during the transcription of the URAT1 gene, including the mRNA that is a product of processing of the primary transcript. The URAT1 can be in a cell, for example, a cell in a subject.

[0025] In the present disclosure, the term "complementary" when used to describe a first nucleobase or nucleotide sequence (e.g., an RNAi agent sense / antisense strand or a targeted mRNA) relative to a second nucleobase or nucleotide sequence refers to the ability of an oligonucleotide or polynucleotide comprising the first nucleotide sequence to hybridize (form base-pair hydrogen bonds under mammalian physiological conditions (or in vitro analogous conditions)) to an oligonucleotide or polynucleotide comprising the second nucleotide sequence and form a duplex or double helix structure under certain standard conditions.

[0026] In the present disclosure, the term "fully complementary" refers to a situation in which, in a pair of hybridized nucleobase or nucleotide sequence molecules, all (100%) of the bases in a contiguous sequence of a first oligonucleotide will hybridize with the same number of bases in a contiguous sequence of a second oligonucleotide. The contiguous sequence can comprise all or a portion of the first or second nucleotide sequence. "Partially complementary" refers to a situation in which, in a pair of hybridized nucleobase or nucleotide sequence molecules, a majority, e.g., at least 70% (but not all) of the bases in a contiguous sequence of a first oligonucleotide will hybridize with the same number of bases in a contiguous sequence of a second oligonucleotide. The contiguous sequence can comprise all or a portion of the first or second nucleotide sequence.

[0027] In the present disclosure, the term "substantially complementary" refers to a situation in which, in a pair of hybridized nucleobase or nucleotide sequence molecules, a substantial majority, e.g., at least 85% or 3 or fewer nucleotide differences (but not all) of the bases in a contiguous sequence of a first oligonucleotide will hybridize with the same number of bases in a contiguous sequence of a second oligonucleotide. The contiguous sequence can comprise all or a portion of the first or second nucleotide sequence.

[0028] In the present disclosure, the terms "complementary," "fully complementary," "partially complementary," and "substantially complementary" are used in reference to nucleobase or nucleotide matches between the sense and antisense strands of an RNAi agent, or between the antisense strand of an RNAi agent and the sequence of a target mRNA.

[0029] The term "complementary" refers to the ability of an oligonucleotide of a first sequence to hybridize and form a duplex structure with an oligonucleotide of a second sequence under certain conditions, unless otherwise specified. "At least partially complementary" means that the two sequences can be fully complementary, or have no more than 5, 4, 3, or 2 mismatched base pairs overall, while retaining the ability to hybridize under the relevant conditions. Additionally, where one or more single-stranded overhangs are designed into the two oligonucleotides when they are designed to hybridize, such overhangs should not be considered mismatches for purposes of determining complementarity. Correspondingly, in the present invention, "mismatch" refers to the situation in which the bases at the corresponding positions in an siRNA duplex molecule are not present in complementary form, unless otherwise specified.

[0030] In the present disclosure, the term "nucleotide difference" and the term "nucleotide base difference" and the term "difference in nucleotide sequence" can be used interchangeably. It means that the base species of a nucleotide at the same or corresponding position has been changed compared to the original nucleotide sequence. For example, when one nucleotide base in the original nucleotide sequence is A, a difference in nucleotide sequence at the same or corresponding position is considered to exist in the case where the nucleotide base is changed to U, C, G, or dT, dC, dG, etc. It should be noted here that in the case where the nucleotide at the same or corresponding position is only different in terms of the presence or absence of a modification or the type of modification compared to the original nucleotide sequence, a difference in nucleotide sequence at the position is not considered to exist.

[0031] In the present disclosure, the term "ligand" or "conjugate group" refers to an atom or group of atoms that binds to an oligonucleotide or other oligomer. Generally, a conjugate group modifies one or more properties of the compound to which it is attached, including, but not limited to, pharmacodynamics, pharmacokinetics, binding, absorption, cellular distribution, cellular uptake, charge, and / or clearance properties. The term "linked" as used herein when referring to the linkage between two molecules means that the two molecules are directly or indirectly linked by a covalent bond or that the two molecules are associated via a non-covalent bond (e.g., a hydrogen bond or an ionic bond).

[0032] In the present disclosure, the term "linked" or "conjugated" when referring to the linkage between two compounds or molecules means that the two molecules are linked by a covalent bond or are associated via a non-covalent bond (e.g., a hydrogen bond or an ionic bond). Unless otherwise specified, the terms "linked" and "conjugated" as used in the present disclosure can refer to the linkage between a first compound and a second compound, with or without any intervening atoms or groups of atoms.

[0033] In the present disclosure, a linking group is one or more atoms that link one molecule or part of a molecule to a second molecule or second part of a molecule. A linking group can comprise any number of atoms or functional groups. In some embodiments, a linking group is used only to link two biologically active molecules.

[0034] Unless otherwise specified, the symbols as used in the present disclosure mean that any group or groups can be attached thereto, which is consistent with the scope of the invention as described in the present disclosure.

[0035] According to the common knowledge in the art, a peptide is a compound which is produced by the linkage of two or more amino acids by amide bonds. Here, the individual amino acids are linked in a certain order (sequence) into a chain. An amino acid is a compound which carries at least one amino group and at least one carboxyl group. Both natural (in vivo protein- generating amino acids), unnatural amino acids or prepared amino acids which can exist in an organism are included.

[0036] In the present disclosure, unless otherwise specified, amino acid residues are in the L-form.

[0037] As used in the present disclosure, the term "standard amino acid" refers to the following twenty amino acids: alanine, arginine, asparagine, aspartic acid (aspartate), cysteine, glutamine, glutamic acid (glutamate), glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine.

[0038] As used in the present disclosure, the term "non-standard amino acid" refers to an amino acid other than a "standard amino acid" as defined in the present disclosure. "Non-standard amino acids" include, but are not limited to, N-formylmethionine, hydroxyproline, selenomethionine, isovaline, citrulline (Cit), ornithine, a-methyl-aspartate (aMeD), a-methyl-leucine (aMeL), N-methylalanine, N-methyl-glycine (NMe G), N-methyl-leucine (NMe L), O-cyclohexyl-alanine (Cha), N-ethylalanine, N,N-ε-dimethyllysine (K(Me)2), methylarginine (Arg(Me)), dimethylarginine (R(Me)2), n-alkylated L-a amino acids, and other amino acid analogs or mimetics that function in a similar manner to naturally occurring amino acids.

[0039] As used in the present disclosure and as understood by one of skill in the art, a polyethylene glycol (PEG) unit refers to a repeating unit of the formula (CH2CH2O). It will be understood that in the chemical structures disclosed in the present disclosure, a PEG unit can be depicted as (CH2CH2O), (OCH2CH2), or (CH2OCH2). It will further be understood that the number indicating the number of repeating PEG units can be placed on either side of the parentheses indicating the PEG unit. It will further be understood that a terminal PEG unit can be capped with an atom (e.g., a hydrogen atom) or some other moiety.

[0040] In the present disclosure, the term "cyclic peptide" indicates that two cysteines in a polypeptide chain are linked by a disulfide bond to form a cyclic polypeptide chain. In the present disclosure, the disulfide bond between the two cysteines in the "cyclic peptide" is indicated by .

[0041] In the present disclosure, in a "2',2'-[halogen, halogen] disubstituted modified nucleotide," both the 2'-OH and the 2'-H of the ribose ring are substituted with a halogen (e.g., F, Cl, Br, or I, preferably F); for example, the structure of a 2',2'-[F,F] disubstituted modified nucleotide is

[0042] In the present disclosure, in a "2',2'-[halogen, C1-6 a ribose ring 2'-OH is replaced by a halogen, a ribose ring 2'-H is replaced by a C 1-6 alkyl group; for example, a 2', 2'-[F, CH3] di-substituted modified nucleotide has the structure wherein Base represents a nucleobase selected from A, U, G, C, or T.

[0043] In the present disclosure, the structure of a "phosphorothioate internucleotide linkage" is the structure of a "phosphorodithioate internucleotide linkage" is

[0044] In the present disclosure, the term "pharmaceutical composition" or "composition" can refer to the use for the treatment of a disease, as well as for in vitro culture experiments of cells. When used for the treatment of a disease, the term "pharmaceutical composition" generally refers to a unit dosage form and can be prepared by any one of the methods well known in the pharmaceutical art. All methods include the step of bringing the active ingredient into association with the carrier that constitutes one or more accessory ingredients. Typically, a composition is prepared by uniformly and intimately bringing the active siRNA into association with a liquid carrier, a finely divided solid carrier, or both. The active ingredient is typically mixed with a solid pharmaceutical carrier, or in the case of liquid oral suspensions, with a liquid pharmaceutical carrier. In either case, the resulting compositions can contain one or more members of a group of excipients, which are pharmaceutically acceptable and compatible with the active ingredient.

[0045] In the present disclosure, the term "pharmaceutically acceptable" means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients of the formulation and / or the mammal being treated with it. Preferably, the "pharmaceutically acceptable" of the present disclosure means approved or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0046] In the present disclosure, the term "pharmaceutically acceptable carrier or excipient" can include any solvent, solid excipient, diluent, or other liquid excipient, and the like, suitable for use in the particular target dosage form. Except insofar as any conventional excipient is incompatible with the siRNA of the present disclosure, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutically acceptable composition, its use is contemplated to be within the scope of this disclosure.

[0047] In the present disclosure, the term "subject" refers to any animal that is the object of examination, research, or treatment, and is not intended to be limited to any particular type of subject. In some embodiments, a human is the preferred subject. In other embodiments, a non-human animal is the preferred subject, including, but not limited to, mice, monkeys, ferrets, cows, sheep, goats, pigs, chickens, turkeys, dogs, cats, horses, and reptiles. In other embodiments, a cell is the preferred subject.

[0048] In the present disclosure, the terms "treatment," "treat," or "treating" can be used interchangeably herein. These terms refer to methods of obtaining beneficial or desired results, including but not limited to therapeutic benefit. By "therapeutic benefit" is meant eradication or amelioration of the underlying disorder being treated. Herein, a therapeutic benefit is achieved with regard to the underlying disorder by observing an improvement in one or more physiological symptoms associated with the disorder in a subject, even though the subject can still be afflicted with the disorder.

[0049] In the present disclosure, the terms "prevention" and "prevent" are used interchangeably and refer to methods of obtaining beneficial or desired results, including but not limited to prophylactic benefit. To obtain "prophylactic benefit," a conjugate, RNAi agent, or composition can be administered to a subject at risk of developing a particular disease, or to a subject reporting one or more physiological symptoms of a disease, even though a diagnosis of this disease can not have been made.

[0050] In the present disclosure, the term "administration" generally refers to introducing or delivering a pharmaceutical preparation of the present disclosure into the body of a subject by any route of introduction or delivery. Any method known to those of skill in the art for contacting a cell, organ, or tissue with the pharmaceutical can be employed. The administration can include, without limitation, intravenous, intraarterial, intranasal, intraabdominal, intramuscular, subcutaneous, or oral. The daily dose can be divided into one, two, or more doses of the appropriate form for administration at one, two, or more times during a certain period of time.

[0051] As used herein, the term "modulating gene expression" means that the expression of a gene, or the level of an RNA molecule or equivalent RNA molecule encoding one or more proteins or protein subunits, is up- or down-regulated such that the expression, level, or activity is greater or less than that observed in the absence of the modulating agent. For example, the term "modulating" can mean "inhibiting," although the use of the term "modulating" is not limited to this definition.

[0052] In the present disclosure, the term "inhibiting" refers to a situation where the expression of a target gene is down-regulated due to siRNA-mediated degradation of the mRNA of the target gene, unless specifically stated otherwise. By "down-regulation" is meant a decrease in the expression level of the target gene by 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% or more relative to the expression level in the absence of siRNA treatment.

[0053] In the present context, the term "inhibiting expression of a URAT1 gene" includes inhibition of a URAT1 gene at any level, e.g., at least partial inhibition of URAT1 gene expression, such as inhibition of at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%. Wherein the expression of a URAT1 gene can be assessed based on the level of any variable associated with URAT1 gene expression, e.g., mRNA level or protein level of URAT1. Inhibition can be assessed by a decrease in the absolute or relative level of one or more of these variables compared to a control level. The control level can be any type of control level utilized in the art, e.g., a pre-dose baseline level, or a level determined from a similar subject, cell, or sample that has never been treated or has been treated with a control, such as a control with only a buffer or an inactive agent.

[0054] The use of any conventional excipients, in addition to any ranges incompatible with the siRNA of the present disclosure, e.g., any adverse biological effects produced or interactions with any other components of a pharmaceutically acceptable composition produced in a deleterious manner, are also contemplated by the present disclosure.

[0055] In the present disclosure, the ratio of the reagents used in the present disclosure is calculated by volume ratio (v / v) unless otherwise specified.

[0056] For the purposes of the present application, the technical solutions and advantages will be more apparent from the following further detailed description of the embodiments of the present application, combined with the examples.

[0057] Double-stranded oligonucleotide:

[0058] In a first aspect of the present disclosure, the present disclosure provides a double-stranded oligonucleotide for inhibiting expression of a URAT1 gene, the double-stranded oligonucleotide comprising a sense strand and an antisense strand, the sense strand and the antisense strand each having a length of 17-30 nucleotides, and the sense strand being at least partially complementary to the antisense strand.

[0059] In some embodiments, the antisense strand has a length of 21-23 nucleotides; the sense strand has a length of 19-21 nucleotides. For example, the antisense strand has a length of 21 nucleotides, and the sense strand has a length of 19 nucleotides.

[0060] wherein the antisense strand comprises at least 17 consecutive nucleotides differing no more than 3 nucleotides from nucleotides 1-19 of any of the nucleotide sequences recited in any one of SEQ ID NOs. 70-138, 287-296, counting from the 5' end.

[0061] In some alternative embodiments, the antisense strand comprises at least 17 consecutive nucleotides differing no more than 3 nucleotides from any of the nucleotide sequences recited in any one of SEQ ID NOs. 70-138, 287-296, counting from the 5' end.

[0062] In some alternative embodiments, the antisense strand comprises at least 17 consecutive nucleotides differing no more than 2 nucleotides from any of the nucleotide sequences recited in any one of SEQ ID NOs. 70-138, 287-296, counting from the 5' end.

[0063] In some alternative embodiments, the antisense strand comprises at least 17 consecutive nucleotides differing no more than 1 nucleotide from any of the nucleotide sequences recited in any one of SEQ ID NOs. 70-138, 287-296, counting from the 5' end.

[0064] In some alternative embodiments, the antisense strand comprises at least 17 consecutive nucleotides selected from nucleotides 1-19 of any of the nucleotide sequences recited in any one of SEQ ID NOs. 70-138, 287-296, counting from the 5' end.

[0065] In some embodiments, the antisense strand is selected from any of the nucleotide sequences recited in any one of SEQ ID NOs. 70-138, 287-296.

[0066] In some alternative embodiments, the sense strand comprises at least 17 consecutive nucleotides differing no more than 3 nucleotides from any of the nucleotide sequences recited in any one of SEQ ID NOs. 1-69, 277-286.

[0067] In some alternative embodiments, the sense strand comprises at least 17 consecutive nucleotides differing no more than 3 nucleotides from any of the nucleotide sequences recited in any one of SEQ ID NOs. 1-69, 277-286.

[0068] In some alternative embodiments, the sense strand comprises at least 17 consecutive nucleotides differing no more than 2 nucleotides from any of the nucleotide sequences recited in any one of SEQ ID NOs. 1-69, 277-286.

[0069] In some alternative embodiments, the sense strand comprises at least 17 consecutive nucleotides differing no more than 1 nucleotide from any of the nucleotide sequences recited in any one of SEQ ID NOs. 1-69, 277-286.

[0070] In some specific embodiments, the positive strand is selected from any of the nucleotide sequences shown in SEQ ID NO. 1-69, 277-286.

[0071] In some alternative implementations, the sense strand and the antisense strand have a mismatch of no more than 3 nucleotides.

[0072] In some alternative implementations, the sense strand and antisense strand have a mismatch of no more than 2 nucleotides.

[0073] In some alternative implementations, the sense strand and the antisense strand have a mismatch of no more than one nucleotide.

[0074] In some specific implementations, the justice chain and the antisense chain are completely complementary.

[0075] In some alternative embodiments, the double-stranded oligonucleotide comprises the sense strand sequence shown in any one of groups RN591001-RN591079, and the 1st to 19th nucleotide sequences of the antisense strand sequence shown in any one of groups RN591001-RN591079, starting from the 5' end:

[0076] In some specific embodiments, the double-stranded oligonucleotide is selected from any one of the groups RN591070-RN591079.

[0077] In some other specific embodiments, the double-stranded oligonucleotide is selected from any one of RN591008, RN591017, RN591023, RN591047, RN591063, RN591066, RN591075, and RN591078.

[0078] In some alternative embodiments, the double-stranded oligonucleotide contains at least one modified nucleotide. In other alternative embodiments, each nucleotide in the double-stranded oligonucleotide is selected from modified nucleotides.

[0079] In some alternative embodiments, the modified nucleotide includes 2'-halogenated nucleotides (e.g., 2'-F-modified nucleotides), 2'-deoxy-modified nucleotides, and 2'-OC-modified nucleotides. 1-6 Alkyl-modified nucleotides (e.g., nucleotides modified with 2'-O-CH3), 2'-O-(CH2) n -O-Me modified nucleotides (e.g., nucleotides modified with 2'-O-CH2CH2-O-CH3), nucleotides modified with 2'-amino groups, 2',2'-[halogen, halogen] disubstituted or 2',2'-[halogen, C] disubstituted nucleotides. 1-6at least one of an alkyl di-substituted modified nucleotide, an abasic nucleotide, or a nucleotide-analogue; n is selected from 1 or 2; the nucleotide-analogue includes at least one of a peptide nucleic acid (PNA), a Morpholino (MNA), a bridged nucleic acid (BNA), a locked nucleic acid (LNA), a glycol nucleic acid (GNA), a threose nucleic acid (TNA), or an unlocked nucleic acid (UNA).

[0080] In some alternative embodiments, the modified nucleotide includes at least one of a 2'-F modified nucleotide, a 2'-deoxy modified nucleotide, a 2'-O-CH3 modified nucleotide, a 2'-O-CH2CH2-O-CH3 modified nucleotide, or a 2',2'-[F,CH3] di-substituted modified nucleotide. In other alternative embodiments, the modified nucleotide includes at least one of a 2'-F modified nucleotide, a 2'-O-CH3 modified nucleotide, or a 2'-O-CH2CH2-O-CH3 modified nucleotide.

[0081] In some alternative embodiments, the antisense strand of the double-stranded oligonucleotide is 21 nucleotides in length, the sense strand is 19 nucleotides in length, and each nucleotide in the double-stranded oligonucleotide is a modified nucleotide; wherein:

[0082] In some alternative embodiments, the antisense strand of the double-stranded oligonucleotide is 21 nucleotides in length, the sense strand is 19 nucleotides in length, and each nucleotide in the double-stranded oligonucleotide is a modified nucleotide; wherein:

[0083] In some alternative embodiments, the antisense strand of the double-stranded oligonucleotide is 21 nucleotides in length, the sense strand is 19 nucleotides in length, and each nucleotide in the double-stranded oligonucleotide is a modified nucleotide; wherein:

[0084] In some alternative embodiments, the antisense strand of the double stranded oligonucleotide is 21 nucleotides in length, the sense strand is 19 nucleotides in length, and each nucleotide in the double stranded oligonucleotide is a modified nucleotide; wherein:

[0085] In some alternative embodiments, the antisense strand of the double stranded oligonucleotide is 21 nucleotides in length, the sense strand is 19 nucleotides in length, and each nucleotide in the double stranded oligonucleotide is a modified nucleotide; wherein:

[0086] In some alternative embodiments, the antisense strand of the double stranded oligonucleotide is 21 nucleotides in length, the sense strand is 19 nucleotides in length, and each nucleotide in the double stranded oligonucleotide is a modified nucleotide; wherein:

[0087] In some alternative embodiments, the antisense strand of the double stranded oligonucleotide is 21 nucleotides in length, the sense strand is 19 nucleotides in length, and each nucleotide in the double stranded oligonucleotide is a modified nucleotide; wherein:

[0088] In some alternative embodiments, the antisense strand of the double stranded oligonucleotide is 21 nucleotides in length, the sense strand is 19 nucleotides in length, and each nucleotide in the double stranded oligonucleotide is a modified nucleotide; wherein:

[0089] In some embodiments, the antisense strand of the double stranded oligonucleotide is 21 nucleotides in length, the sense strand is 19 nucleotides in length, and each nucleotide in the double stranded oligonucleotide is a modified nucleotide; wherein:

[0090] In the 5' end to 3' end direction, the 2nd, 6th, 9th, 14th, and 16th nucleotides of the antisense strand are selected from 2'-F modified nucleotides, the 15th nucleotide is selected from a 2'-0-CH2CH2-0-CH3 modified nucleotide, and the remaining nucleotides are 2'-0-CH3 modified nucleotides; the 7th through 10th nucleotides of the sense strand are selected from 2'-F modified nucleotides, and the remaining nucleotides are selected from 2'-0-CH3 modified nucleotides.

[0091] In some alternative embodiments, the double stranded oligonucleotide has at least one modified internucleoside linkage selected from a phosphorothioate internucleoside linkage, and the phosphorothioate internucleoside linkage is present at at least one of positions 1) - 5):

[0092] 1) between the 1st and 2nd nucleotides, and / or between the 2nd and 3rd nucleotides of the sense strand, counting from the 5' terminus;

[0093] 2) between the 1st and 2nd nucleotides, and / or between the 2nd and 3rd nucleotides of the sense strand, counting from the 3' terminus;

[0094] 3) between the 1st and 2nd nucleotides, and / or between the 2nd and 3rd nucleotides of the antisense strand, counting from the 5' terminus;

[0095] 4) between the 1st and 2nd nucleotides, and / or between the 2nd and 3rd nucleotides, and / or between the 3rd and 4th nucleotides of the antisense strand, counting from the 3' terminus;

[0096] 5) between the 10th and 11th nucleotides of the antisense strand, counting from the 5' terminus.

[0097] In some alternative embodiments, each nucleotide in the double stranded oligonucleotide is a modified nucleotide.

[0098] In some alternative embodiments, in the 5' end to 3' end direction, at least four of the nucleotides at positions 2, 6, 9, 12, 14, 16 of the antisense strand are selected from 2'-F modified nucleotides, each of the remaining nucleotide positions other than the 2'-F modified nucleotides is independently selected from a 2'-0-CH3 modified nucleotide or a 2'-0-CH2CH2-0-CH3 modified nucleotide; at least three of the nucleotides at positions 7-10 of the sense strand are selected from 2'-F modified nucleotides, the remaining nucleotide position other than the 2'-F modified nucleotides is selected from a 2'-0-CH3 modified nucleotide.

[0099] In some alternative embodiments, in the 5' end to 3' end direction, the nucleotides at positions 2, 6, 14, 16 of the antisense strand are selected from 2'-F modified nucleotides, any one of the nucleotides at positions 9 and 12 is selected from a 2'-F modified nucleotide, the other nucleotide is selected from a 2'-0-CH3 modified nucleotide, the nucleotide at position 15 is selected from a 2'-0-CH2CH2-0-CH3 modified nucleotide or a 2'-0-CH3 modified nucleotide, and the remaining nucleotides are 2'-0-CH3 modified nucleotides; the nucleotides at positions 7-10 of the sense strand are selected from 2'-F modified nucleotides, and the remaining nucleotides are selected from 2'-0-CH3 modified nucleotides.

[0100] In some alternative embodiments, in the 5' end to 3' end direction, the nucleotides at positions 2, 6, 14, 16 of the antisense strand are selected from 2'-F modified nucleotides, any one of the nucleotides at positions 9 and 12 is selected from a 2'-F modified nucleotide, the other nucleotide is selected from a 2'-0-CH3 modified nucleotide, and the remaining nucleotides are 2'-0-CH3 modified nucleotides; the nucleotides at positions 7-10 of the sense strand are selected from 2'-F modified nucleotides, and the remaining nucleotides are selected from 2'-0-CH3 modified nucleotides.

[0101] In some alternative embodiments, in the 5' end to 3' end direction, the nucleotides at positions 2, 6, 9, 14, 16 of the antisense strand are selected from 2'-F modified nucleotides, and the remaining nucleotides are 2'-0-CH3 modified nucleotides; the nucleotides at positions 7-10 of the sense strand are selected from 2'-F modified nucleotides, and the remaining nucleotides are selected from 2'-0-CH3 modified nucleotides.

[0102] In other optional embodiments, the 2nd, 6th, 12th, 14th, 16th nucleotide of the antisense strand, in the 5' end to 3' end direction, is selected from a 2'-F modified nucleotide, and the nucleotides at the remaining positions are 2'-0-CH3 modified nucleotides; and the 7th-10th nucleotides of the sense strand are selected from 2'-F modified nucleotides, and the nucleotides at the remaining positions are selected from 2'-0-CH3 modified nucleotides.

[0103] In some optional embodiments, the double stranded oligonucleotide further has at least one modified internucleoside linkage (e.g., 1, 2, 3, 4, 5, 6, 7, or 8, etc.), the modified internucleoside linkage comprising at least one of a phosphorothioate internucleoside linkage or a phosphorodithioate internucleoside linkage, and at least one of the modified internucleoside linkages is each independently selected from between the 1stand 2ndnucleotides of the sense strand starting from the 5' end, between the 2ndand 3rdnucleotides of the sense strand starting from the 5' end, between the 1stand 2ndnucleotides of the sense strand starting from the 3' end, between the 1stand 2ndnucleotides of the antisense strand starting from the 5' end, between the 2ndand 3rdnucleotides of the antisense strand starting from the 5' end, between the 10thand 11thnucleotides of the antisense strand starting from the 5' end, between the 1stand 2ndnucleotides of the antisense strand starting from the 3' end, or between the 2ndand 3rdnucleotides of the antisense strand starting from the 3' end.

[0104] In some optional embodiments, the modified internucleoside linkage is selected from a phosphorothioate internucleoside linkage.

[0105] In some optional embodiments, the double stranded oligonucleotide has at least 6 modified internucleoside linkages (e.g., phosphorothioate internucleoside linkages), wherein the 6 modified internucleoside linkages are selected from any 6 of between the 1stand 2ndnucleotides of the sense strand starting from the 5' end, or between the 2ndand 3rdnucleotides of the sense strand starting from the 5' end, or between the 1stand 2ndnucleotides of the sense strand starting from the 3' end, or between the 1stand 2ndnucleotides of the antisense strand starting from the 5' end, or between the 2ndand 3rdnucleotides of the antisense strand starting from the 5' end, or between the 10thand 11thnucleotides of the antisense strand starting from the 5' end, or between the 1stand 2ndnucleotides of the antisense strand starting from the 3' end, or between the 2ndand 3rdnucleotides of the antisense strand starting from the 3' end.

[0106] In some embodiments, the double stranded oligonucleotide has at least 6 modified internucleotide linkages (e.g., phosphorothioate internucleotide linkages), wherein the 6 modified internucleotide linkages are located between the 1stand 2ndnucleotides from the 5' terminus of the sense strand, between the 2ndand 3rdnucleotides from the 5' terminus of the sense strand, between the 1stand 2ndnucleotides from the 5' terminus of the antisense strand, between the 2ndand 3rdnucleotides from the 5' terminus of the antisense strand, between the 1stand 2ndnucleotides from the 3' terminus of the antisense strand, and between the 2ndand 3rdnucleotides from the 3' terminus of the antisense strand, respectively.

[0107] In other alternative embodiments, the internucleotide linkage between the 10thand 11thnucleotides from the 5' terminus of the antisense strand is selected from a modified internucleotide linkage (e.g., a phosphorothioate internucleotide linkage), and the internucleotide linkages between the nucleotide of the sense strand that base pairs with the 10thnucleotide of the antisense strand and the nucleotide of the sense strand that base pairs with the 11thnucleotide of the antisense strand are selected from phosphate internucleotide linkages.

[0108] In other alternative embodiments, the double stranded oligonucleotide has one phosphorodithioate internucleotide linkage, and the phosphorodithioate internucleotide linkage is located between the 1stand 2ndnucleotides from the 5' terminus of the sense strand, or between the 2ndand 3rdnucleotides from the 5' terminus of the sense strand, or between the 1stand 2ndnucleotides from the 3' terminus of the sense strand, or between the 1stand 2ndnucleotides from the 5' terminus of the antisense strand, or between the 2ndand 3rdnucleotides from the 5' terminus of the antisense strand, or between the 10thand 11thnucleotides from the 5' terminus of the antisense strand, or between the 1stand 2ndnucleotides from the 3' terminus of the antisense strand, or between the 2ndand 3rdnucleotides from the 3' terminus of the antisense strand.

[0109] In some alternative embodiments, the 1stnucleotide from the 5' terminus of the antisense strand is selected from or a 5'-phosphate analog modified nucleotide; the 5'-phosphate analog modified nucleotide is selected from any one of the nucleotides represented by the following structures:

[0110] wherein Base represents a nucleobase selected from A, U, G, C, or T; R represents -H, -OH, -CH3, -OCH3, -F.

[0111] In some alternative embodiments, the 5'-phosphate analog modified nucleotide is selected from a 5'-(E)-vinyl phosphonate (5'-(E)-VP) modified nucleotide.

[0112] In some alternative embodiments, the 1st nucleotide of the antisense strand, counting from the 5' terminus, is selected from or the 1st nucleotide of the antisense strand, counting from the 5' terminus, is selected from 5'-(E)-VP modified nucleotides. In some particular embodiments, the 1st nucleotide of the antisense strand, counting from the 5' terminus, is selected from In other particular embodiments, the 1st nucleotide of the antisense strand, counting from the 5' terminus, is selected from 5'-(E)-VP modified nucleotides.

[0113] In some alternative embodiments, the antisense strand comprises, counting from the 5' terminus, at least 17 consecutive nucleotides differing by no more than 3 nucleotides from the 1st-19th nucleotides of the nucleotide sequence set forth in any one of SEQ ID NO. 208-276, 307-316.

[0114] In some alternative embodiments, the antisense strand comprises, counting from the 5' terminus, at least 17 consecutive nucleotides differing by no more than 3 nucleotides from the 1st-19th nucleotides of the nucleotide sequence set forth in any one of SEQ ID NO. 208-276, 307-316.

[0115] In some alternative embodiments, the antisense strand comprises, counting from the 5' terminus, at least 17 consecutive nucleotides differing by no more than 2 nucleotides from the 1st-19th nucleotides of the nucleotide sequence set forth in any one of SEQ ID NO. 208-276, 307-316.

[0116] In some alternative embodiments, the antisense strand comprises, counting from the 5' terminus, at least 17 consecutive nucleotides differing by no more than 1 nucleotide from the 1st-19th nucleotides of the nucleotide sequence set forth in any one of SEQ ID NO. 208-276, 307-316.

[0117] In some alternative embodiments, the antisense strand comprises, counting from the 5' terminus, the 1st-19th nucleotides of the nucleotide sequence set forth in any one of SEQ ID NO. 208-276, 307-316;

[0118] In some particular embodiments, the antisense strand is selected from the nucleotide sequence set forth in any one of SEQ ID NO. 208-276, 307-316.

[0119] In some alternative embodiments, the sense strand comprises, at least 17 consecutive nucleotides differing by no more than 3 nucleotides from the nucleotide sequence set forth in any one of SEQ ID NO. 139-207, 297-306.

[0120] In some alternative embodiments, the sense strand comprises no more than 3 nucleotides different from the nucleotide sequence set forth in any one of SEQ ID NO. 139-207, 297-306.

[0121] In some alternative embodiments, the sense strand comprises no more than 2 nucleotides different from the nucleotide sequence set forth in any one of SEQ ID NO. 139-207, 297-306.

[0122] In some alternative embodiments, the sense strand comprises no more than 1 nucleotide different from the nucleotide sequence set forth in any one of SEQ ID NO. 139-207, 297-306.

[0123] In some specific embodiments, the sense strand is selected from the nucleotide sequence set forth in any one of SEQ ID NO. 139-207, 297-306.

[0124] In some alternative embodiments, the double-stranded oligonucleotide is selected from the sense strand sequence set forth in any one of the group RX591070-RX591079, and the 1st-19th nucleotide sequence, counting from the 5' end, of the antisense strand sequence set forth in any one of the group RX591070-RX591079:

[0125] In some specific embodiments, the double-stranded oligonucleotide is selected from any one of the group RX591001-RX591079.

[0126] In some specific embodiments, the double-stranded oligonucleotide is selected from any one of the group RX591008, RX591017, RX591023, RX591047, RX591063, RX591066, RX591075, RX591078.

[0127] Conjugate:

[0128] In a second aspect of the present disclosure, the present disclosure provides a conjugate comprising the double-stranded oligonucleotide of the first aspect of the present disclosure and one or more targeting ligand groups conjugated to the double-stranded oligonucleotide.

[0129] In some alternative embodiments, the targeting ligand group targets the kidney Megalin receptor or the kidney natriuretic peptide receptor.

[0130] In some alternative embodiments, the targeting ligand group is selected from a polypeptide fragment comprising at least 4 amino acids.

[0131] In some alternative embodiments, the targeting ligand group comprises any one of the polypeptide fragments shown in 1) - 6) below:

[0132] 1) - X1(KKEEE) n - K m - wherein n = 1 - 5, m = 0 or 1 or 2, X1is selected from any L-alpha amino acid or L-beta amino acid except K, E;

[0133] 2) - CLPVASC-, preferably in a cyclic peptide form

[0134] 3) - CYFQNC-, preferably in a cyclic peptide form

[0135] 4) - KIDRI-;

[0136] 5) - IDRI- (Ile-Asp-Arg-Ile-);

[0137] 6) - dXa-Ser-dXb-X2-dXc-Gly-Xd-Ile-Asp-Arg(Ak)-Ile-; wherein X2is selected from any unnatural amino acid; dXa, dXb, dXc are selected from any D-amino acid; Arg(Ak) is selected from arginine or alkylated modified arginine; Xdis selected from Hyp or Pro.

[0138] In some alternative embodiments, the targeting ligand group is selected from a polypeptide comprising any one of the amino acid sequence segments shown in a) - g):

[0139] a) - KKEEE-KKEEE-KKEEE-K-;

[0140] b) - CKKEEE-KKEEE-KKEEE-K;

[0141] c) - CLPVASC-, preferably in a cyclic peptide form

[0142] d) - KIDRI-;

[0143] e) - CYFQNCPRG-, preferably in a cyclic peptide form

[0144] f) - DPhe-Ser-DHyp-Cha-DAla-Gly-Hyp-Ile-Asp-Arg(Me)-Ile-;

[0145] g) - DPhe-Ser-DPro-Cha-DAla-Gly-Pro-Ile-Asp-Arg-Ile-.

[0146] In some alternative embodiments, the targeting ligand group is linked to the double-stranded oligonucleotide via a linker;

[0147] Optionally, the structural unit of the targeting ligand group-linker in the conjugate is selected from any of the structures shown in 1) - 7) below:

[0148] 1) La-KKEEE-KKEEE-KKEEE-K-Laa;

[0149] 2) La-CKKEEE-KKEEE-KKEEE-K-Laa;

[0150] 3) La-CLPVASC-Laa, preferably in cyclic peptide form

[0151] 4) La-DPhe-Ser-DHyp-Cha-DAla-Gly-Hyp-Ile-Asp-Arg(Me)-Ile-Laa;

[0152] 5) La-CYFQNCPRG-Laa; preferably in cyclic peptide form

[0153] 6) La-DPhe-Ser-DPro-Cha-DAla-Gly-Pro-Ile-Asp-Arg-Ile-Laa;

[0154] 7) Lb-KIDRI-Laa;

[0155] In formulas 1) - 6), either La or Laa is a linker and the other is a terminal blocking group;

[0156] In formula 7), either Lb or Laa is a linker and the other is a terminal blocking group;

[0157] The linker is a linking group comprising at least one of a triazole group or a PEG unit or an acyl group.

[0158] In some alternative embodiments, in formulas 1) - 6), La is a linker and Laa is a terminal blocking group, La is selected from a linking group comprising -PEG2-CH2CH2CO- or -triazole-PEG2-; and Laa is selected from an amino group, or an alkyl-substituted amine group such as -NHCH3.

[0159] In some alternative embodiments, in formula 7), Lb is a linker and Laa is a terminal blocking group, Lb is selected from a linking group comprising a 6-10 membered aromatic or heteroaromatic ring; and Laa is selected from an amino group, or an alkyl-substituted amine group such as -NHCH3.

[0160] In some optional embodiments, the targeting ligand group is attached to the end of the sense strand and or the antisense strand of the double stranded oligonucleotide (e.g. the 3’ end of the sense strand, and / or the 5’ end of the sense strand, and / or the 3’ end of the sense strand) via a linker.

[0161] In some optional embodiments, the structure of the targeting ligand group-linker in the conjugate is selected from any of the structures shown in a) - j), or a pharmaceutically acceptable salt thereof:

[0162] a) -La-KKEEE-KKEEE-KKEEE-K-Lc;

[0163] b) -La-CKKEEE-KKEEE-KKEEE-K-Lc;

[0164] c) -La-CLPVASC-Lc; preferably in the form of a cyclic peptide

[0165] d) -La-DPhe-Ser-DHyp-Cha-DAla-Gly-Hyp-Ile-Asp-Arg(Me)-Ile-Lc;

[0166] e) -La-CYFQNCPRG-Lc; preferably in the form of a cyclic peptide

[0167] f) -La-DPhe-Ser-DPro-Cha-DAla-Gly-Pro-Ile-Asp-Arg-Ile-Lc;

[0168] g)

[0169] h)

[0170] i)

[0171] j)

[0172] wherein La is a linker independently selected from any bond, -NH-, an amido group or a linking group comprising at least one PEG unit; and Lc is a terminal blocking group.

[0173] In some optional embodiments, La is a linking group comprising at least 2 PEG units.

[0174] In some optional embodiments, La is selected from an amido group, or n is 1-3 (e.g. 1, 2, 3), m is 0 or 1, and Z is -CO- (carbonyl) or -NH-. represents a site covalently linked to a double-stranded oligonucleotide, and * represents a site covalently linked to a polypeptide fragment.

[0175] In some alternative embodiments, Lcis a blocking group at the C-terminus of the polypeptide fragment. In other alternative embodiments, Lcis selected from an amino or alkyl substituted amine group (e.g., -NHCH3).

[0176] Pharmaceutical composition:

[0177] In a third aspect of the present disclosure, the present disclosure provides a pharmaceutical composition comprising the double-stranded oligonucleotide of the first aspect of the present disclosure, and / or the conjugate of the second aspect of the present disclosure.

[0178] In some alternative embodiments, the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers or excipients.

[0179] Use in the manufacture of a medicament for inhibiting the expression of a URAT1 gene:

[0180] In a fourth aspect of the present disclosure, the present disclosure provides use of the double-stranded oligonucleotide of the first aspect of the present disclosure, and / or the conjugate of the second aspect of the present disclosure, and / or the pharmaceutical composition of the third aspect of the present disclosure in the manufacture of a medicament for inhibiting the expression of a URAT1 gene.

[0181] Use in the manufacture of a medicament for treating and / or preventing a pathological condition or disease associated with the expression of URAT1:

[0182] In a fifth aspect of the present disclosure, the present disclosure provides use of the double-stranded oligonucleotide of the first aspect of the present disclosure, and / or the conjugate of the second aspect of the present disclosure, and / or the pharmaceutical composition of the third aspect of the present disclosure in the manufacture of a medicament for treating and / or preventing a pathological condition or disease associated with the expression of URAT1.

[0183] In some alternative embodiments, the pathological condition or disease is hyperuricemia or gout.

[0184] Method for inhibiting the expression of a URAT1 gene:

[0185] In a sixth aspect of the present disclosure, the present disclosure provides a method for inhibiting the expression of a URAT1 gene, comprising administering to a subject an effective amount of the double-stranded oligonucleotide of the first aspect of the present disclosure, and / or the conjugate of the second aspect of the present disclosure, and / or the pharmaceutical composition of the third aspect of the present disclosure.

[0186] In some alternative embodiments, the subject is a human. In other alternative embodiments, the subject is a cell, such as a Vero cell or a HuH1 cell.

[0187] A method for preventing and / or treating hyperuricemia or gout:

[0188] In a seventh aspect of the present disclosure, the present disclosure provides a method for preventing and / or treating hyperuricemia or gout, comprising administering to a subject an effective dose of the double-stranded oligonucleotide according to the first aspect of the present disclosure, and / or the conjugate according to the second aspect of the present disclosure, and / or the pharmaceutical composition according to the third aspect of the present disclosure.

[0189] In some alternative embodiments, the subject is a human.

[0190] The effective amount of the double-stranded oligonucleotide, double-stranded oligonucleotide conjugate or pharmaceutical composition according to the present disclosure can vary depending on the mode of administration and the severity of the disease to be treated, etc. Specifically, the selection of the effective amount can be determined by a person of ordinary skill in the art (e.g. through clinical trials) according to various factors. The factors include, but are not limited to, the pharmacokinetic parameters of the active ingredient, such as bioavailability, metabolism, half-life, etc.; the severity of the disease to be treated in the patient, the weight of the patient, the immune status of the patient, the route of administration, etc.

[0191] The subject can be administered by any suitable route known in the art, including but not limited to oral or parenteral routes, including intravenous administration, intramuscular administration, subcutaneous administration, transdermal administration, airway administration (aerosol), pulmonary administration, nasal administration, rectal administration and topical administration (including buccal administration and sublingual administration), such as intravenous injection.

[0192] The double-stranded oligonucleotide, double-stranded oligonucleotide conjugate and pharmaceutical composition provided by the present disclosure can effectively inhibit the expression of the URAT1 gene, which is helpful for the treatment and / or prevention of pathological conditions or diseases associated with the expression of URAT1.

[0193] In order to make the purposes, technical solutions and advantages of the present disclosure clearer, the embodiments of the present disclosure will be further described in detail below with reference to the examples.

[0194] Unless otherwise specified, the reagents, reagent consumables and instruments and equipment used in the present disclosure are commercially available. Among them, the main reagent consumables are shown in Table 1, and the main instruments and equipment are shown in Table 2.

[0195] Table 1 Main reagent consumables

[0196] Table 2 Main instruments and equipment

[0197] Preparation Example 1, synthesis of siRNA:

[0198] siRNA sequence design:

[0199] The siRNA sequence of the present disclosure targets all transcripts and predicted transcripts (NM_144585.4, NM_153378.3, NM_001276326.2, NM_001276327.2, XM_006718430.5, XM_006718431.5, XM_054367604.1, XM_054367605.1) of the URAT1 gene, and the sequence information of the URAT1 gene is derived from the NCBI Gene Database (https: / / www.ncbi.nlm.nih.gov / gene / ).

[0200] Synthesis of siRNA:

[0201] (1-1) Synthesis of sense strand (SS) and antisense strand (AS):

[0202] The nucleotide sequence is connected one by one from the 3' end to the 5' end by the method of phosphoramidite nucleic acid solid-phase synthesis, starting with the cycle of the compound linked to the solid-phase carrier (such as CPG carrier, PS carrier) described above.

[0203] Each connection of a nucleotide monomer includes four-step reactions of deprotection, coupling, capping, oxidation or sulfuration. The synthesis conditions are given as follows:

[0204] The nucleotide monomers are prepared into an acetonitrile solution of nucleotide monomers with a concentration of 0.1 M.

[0205] The conditions of deprotection reaction of each step are the same. The conditions of deprotection reaction are: temperature is 25°C, reaction time is 70 seconds, deprotection reagent is dichloroacetic acid in dichloromethane with a concentration of 3% (v / v), and the molar ratio of dichloroacetic acid to 4,4'-dimethoxytrityl protecting group on the solid-phase carrier is 5:1.

[0206] The conditions of coupling reaction of each step are the same. The conditions of coupling reaction are: temperature is 25°C, the molar ratio of the nucleic acid sequence connected on the solid-phase carrier to the nucleotide monomer is 1:10, the molar ratio of the nucleic acid sequence connected on the solid-phase carrier to the coupling reagent is 1:65, reaction time is 600 seconds, the coupling reagent is 5-ethylthio-1H-tetrazole in acetonitrile with a concentration of 0.5 M, and the thio reagent is a mixture of acetonitrile / pyridine with a concentration of 0.2 mol / L (the volume ratio of acetonitrile and pyridine is 1:1).

[0207] The conditions of each step of the capping reaction are the same. The capping reaction conditions are: temperature is 25°C; reaction time is 2 minutes; capping reagent solution is a mixed solution of Cap1 and Cap2. Cap1 is a pyridine / acetonitrile mixed solution of N-methylimidazole with a concentration of 20% (v / v), the volume ratio of pyridine to acetonitrile is 3:5, and Cap2 is an acetic anhydride acetonitrile solution with a concentration of 20% (v / v); the molar ratio of N-methylimidazole in Cap1 capping reagent, acetic anhydride in Cap2 capping reagent, and the nucleic acid sequence connected on the solid support is 1:1:1.

[0208] The conditions of each step of the oxidation reaction are the same. The oxidation reaction conditions are: temperature is 25°C; reaction time is 3 seconds; the concentration of the oxidation reagent is 0.05M iodine water, the molar ratio of iodine to the nucleic acid sequence connected on the solid support in the coupling reaction is 30:1; the oxidation reaction is carried out in a water / pyridine mixed solvent (the volume ratio of water to pyridine is 1:9). The conditions of the sulfurization reaction are: temperature is 25°C; reaction time is 360 seconds; the concentration of the sulfur reagent is 0.2M pyridine solution of hydrogenated xanthan, the molar ratio of the sulfur reagent to the nucleic acid sequence connected on the solid support in the coupling reaction is 4:1; the sulfurization reaction is carried out in a water / pyridine mixed solvent (the volume ratio of water to pyridine is 1:9).

[0209] After the last nucleotide monomer is connected, the nucleic acid sequence connected on the solid support is sequentially subjected to cleavage, deprotection, purification, desalting, and then freeze-drying to obtain the sense strand, wherein:

[0210] The cleavage and deprotection conditions are as follows: the synthesized nucleotide sequence connected with the solid support is added to 25wt% ammonia water, the amount of ammonia water is 0.5ml / μmol, the reaction is carried out at 55°C for 16 hours, the solvent is removed, and vacuum concentration is carried out to dryness. After ammonia water treatment, the product is dissolved with 0.4ml / μmol N-methylpyrrolidine, followed by adding 0.3ml / μmol triethylamine and 0.6ml / μmol triethylamine trifluoride, and the 2'-O-TBDMS protection on the ribose is removed.

[0211] Purification and desalination: the purification of nucleic acids was completed by gradient elution of NaCl using a preparative ion chromatography purification column (Source 15Q). Specifically, eluent 1 was 20 mM sodium phosphate (pH = 8.1) and solvent was a water / acetonitrile mixture (9:1 by volume); eluent 2 was 1.5 M sodium chloride, 20 mM sodium phosphate (pH = 8.1) and solvent was a water / acetonitrile mixture (9:1 by volume); the elution gradient was eluent 1:eluent 2 = (100:0)-(50:50). The product eluate was collected and desalted using a reverse phase chromatography purification column, which included desalting using a dextran gel column with dextran gel G25 and elution with deionized water.

[0212] Detection: purity was detected using ion exchange chromatography (IEX-HPLC); molecular weight was detected using liquid chromatography-mass spectrometry (LC-MS) and the measured value was compared with the theoretical value; if the measured value and the theoretical value were consistent, it was indicated that the target sense strand and antisense strand were obtained.

[0213] (1-2) Synthesis of siRNA:

[0214] The sense strand and antisense strand synthesized in step (1-1) were mixed in an equimolar ratio, dissolved in water for injection and heated to 95°C, slowly cooled to room temperature and kept at room temperature for 10 minutes, so that the sense strand and antisense strand formed a double-stranded structure through hydrogen bonding, thereby obtaining the siRNA shown in Tables 4a and 4b.

[0215] Table 3a, unmodified siRNA nucleotide sequence information:

[0216] Table 4a, modified siRNA sequence information:

[0217] Table 3b, unmodified siRNA sequence information:

[0218] Table 4b, modified siRNA sequence information:

[0219] Unless otherwise specified, the base composition and modification in the disclosure are as follows: capital letters A, U, G, C, T represent the base composition of nucleotides, and lowercase letter m represents that the nucleotide represented by the capital letter adjacent to the left of the letter m is a 2'-O-CH3 modified nucleotide; lowercase letter f represents that the nucleotide represented by the capital letter adjacent to the left of the letter f is a 2'-F modified nucleotide; (moe) represents that the nucleotide represented by the capital letter adjacent to the left of (moe) is a 2'-O-CH2CH2-O-CH3 modified nucleotide; and lowercase letter s represents that the internucleotide bond between the two nucleotides adjacent to the left and right of the letter s is a phosphorothioate bond internucleotide bond.

[0220] The structural formula of the 2'-O-CH3 modified nucleotide is

[0221] The structural formula of the 2'-F modified nucleotide is

[0222] The structural formula of the 2'-O-CH2CH2-O-CH3 modified nucleotide is

[0223] In the formula, Base represents a nucleoside base, and the nucleoside base is selected from A, U, G, C or T. Q represents -OH, -O - , -SH or -S - .

[0224] Unless otherwise specified, the siRNA sequence (modification) used in the disclosure is synthesized by Suzhou Xuanjing Biotechnology Co., Ltd.

[0225] Biological detection experiment

[0226] Unless otherwise specified, the sequence used in the biological detection experiment of the disclosure is a modified siRNA.

[0227] Unless otherwise specified, the PCR primers used in the disclosure are entrusted to Beijing Qikang Biotechnology Co., Ltd. for synthesis.

[0228] Unless otherwise specified, in the real-time fluorescent quantitative PCR method, the expression level and inhibition rate of the target gene mRNA in each test group are relatively quantitatively calculated by using the ΔΔCt method, and the calculation method is as follows: ΔCt(test group) = Ct(target gene in test group) - Ct(internal reference gene in test group) ΔCt(blank control group) = Ct(target gene in blank control group) - Ct(internal reference gene in blank control group) ΔΔCt(test group) = ΔCt(test group) - ΔCt(blank control group average) ΔΔCt(blank control group) = ΔCt(blank control group) - ΔCt(blank control group average)

[0229] Wherein, in the cell experiment, ACt(blank control group average) is the arithmetic mean of several culture hole ACt(blank control group) in the blank control group. Therefore, each sample of the test group and the blank control group corresponds to a ΔΔCt value. Test group target gene mRNA relative expression level = 2-ΔΔCt(test group) x 100%

[0230] The test group target gene mRNA relative expression level is normalized based on the blank control group, and the blank control group target gene mRNA relative expression level is defined as 100%.

[0231] Test group target gene mRNA expression inhibition rate = 100%-test group target gene mRNA relative expression level

[0232] Unless otherwise specified, the experimental data of the inhibitory activity are expressed as X ± STDEV. (X ± STDEV). The experimental data are plotted and analyzed using GraphPad prism 8.0 software.

[0233] Example 1, evaluation of the inhibitory activity of siRNA sequences on the target gene urate transporter 1 (URAT1) in Vero cells:

[0234] This example uses an in vitro cell transfection method to evaluate the inhibitory activity of siRNA sequences on the target gene URAT1 in Vero cells.

[0235] Vero cells (donated by Beijing University of Technology) were cultured in complete medium (DMEM medium containing 10% fetal bovine serum and 1% double antibody) at 5% CO2 and 37°C. After the cell growth density reached 80%, the cells were trypsinized and resuspended. The resuspended cells were adjusted to the density with complete medium and inoculated into a 24-well plate at 0.6 x 10 5 / well. After 24 h of culture, transfection was performed.

[0236] Preparation of siRNA transfection mixture: siRNA was prepared into a 20 μM concentration stock solution with PBS, and then 1.5 μl of siRNA stock solution was taken per well to dilute with 48.5 μl of Opti-MEM medium as a 50 μl siRNA working solution; 3 μl of RNAiMAX transfection reagent was diluted with 47 μl of Opti-MEM medium, incubated at room temperature for 5 minutes, and then mixed with 50 μl of siRNA working solution to prepare a siRNA transfection mixture. The MOCK group was prepared by mixing 50 μl of Opti-MEM medium with 50 μl of transfection reagent working solution.

[0237] The 24-well plate was replaced with 500 μl / well Opti-MEM medium on the day of transfection, and 100 μl of siRNA transfection mixture or MOCK transfection mixture was added to each well, so that the final concentration of siRNA was 50 nM / well.

[0238] After 24 hours of continuous culture, total RNA was extracted according to the chemical extraction method. That is, after removing the culture medium from each well, 1 mL of Trizol solution was added, the cells were lysed by blowing, the lysate was transferred to a 1.5 mL centrifuge tube, 200 μL of chloroform was added, and the mixture was shaken well and then placed at room temperature for 3 min. Centrifugation was performed at 4°C and 12000 rpm for 10 min. 400 μL of supernatant was transferred to a centrifuge tube containing 400 μL of isopropanol, mixed well, and placed at room temperature for 10 min. Centrifugation was performed at 4°C and 12000 rpm for 10 min, and the supernatant was discarded. 1 mL of 75% ethanol was added, and the centrifuge tube was inverted to wash the precipitate. Centrifugation was performed at 4°C and 12000 rpm for 5 min, the supernatant was removed, and the total RNA was obtained after drying at room temperature.

[0239] 1 μg of the total RNA was taken and a reverse transcription kit (Promega, Reverse Transcription System, A3500) was used with an Oligo(dT)15 reverse transcription primer. A 20 μL reverse transcription system was configured according to the method described in the reverse transcription kit manual, and the reverse transcription reaction was completed. After the reaction, 80 μL of RNase-Free water was added to the reverse transcription system to obtain a cDNA solution. Then, a real-time fluorescent quantitative PCR kit (ABI, SYBR Select Master Mix, Catalog number: 4472908) was used to detect the expression of the target gene mRNA in the vero cells. In this real-time fluorescent quantitative PCR method, primers for the target gene and primers for the internal reference gene were used to detect the target gene and the internal reference gene, respectively. According to the method described in the real-time fluorescent quantitative PCR kit manual, a 20 μL Real-time PCR reaction system was configured for each PCR detection well, and each reaction system contained 5 μL of the cDNA solution obtained from the reverse transcription reaction, 10 μL of SYBR Select Master Mix, 0.5 μL of 10 μM upstream primer, 0.5 μL of 10 μM downstream primer, and 4 μL of RNase-Free H2O. TM Select Master Mix, Catalog number: 4472908) was used to detect the expression of the target gene mRNA in the vero cells. In this real-time fluorescent quantitative PCR method, primers for the target gene and primers for the internal reference gene were used to detect the target gene and the internal reference gene, respectively. According to the method described in the real-time fluorescent quantitative PCR kit manual, a 20 μL Real-time PCR reaction system was configured for each PCR detection well, and each reaction system contained 5 μL of the cDNA solution obtained from the reverse transcription reaction, 10 μL of SYBR TM Select Master Mix, Catalog number: 4472908) was used to detect the expression of the target gene mRNA in the vero cells. In this real-time fluorescent quantitative PCR method, primers for the target gene and primers for the internal reference gene were used to detect the target gene and the internal reference gene, respectively. According to the method described in the real-time fluorescent quantitative PCR kit manual, a 20 μL Real-time PCR reaction system was configured for each PCR detection well, and each reaction system contained 5 μL of the cDNA solution obtained from the reverse transcription reaction, 10 μL of SYBR TM) Real-time PCR amplification was performed using a three-step method, and the amplification procedure was 10 min of pre-denaturation at 95°C, followed by 40 cycles of denaturation at 95°C for 30 s, annealing at 60°C for 30 s, and extension at 72°C for 30 s.

[0240] Table 5 Sequence list of primers used in Example 1

[0241] The results of Example 1 show that the designed sequences can effectively reduce the expression of URAT1 mRNA, and the sequences RX591008, RX591011, RX591013, RX591014, RX591017, RX591018, RX591019, RX591022, and RX591024 have relatively better gene inhibition levels (Table 6).

[0242] Table 6 Inhibition activity of Vero cells on target genes after administration of siRNA conjugates described in this example

[0243] Example 2, Evaluation of siRNA sequences on the inhibition activity of target gene URAT1 in HuH1 cells:

[0244] This example uses an in vitro cell transfection method to evaluate the inhibition activity of siRNA sequences on the target gene URAT1 in HuH1 cells.

[0245] HuH1 cells (Wuhan Ponsay Biotechnology Co., Ltd.) were cultured in complete medium (DMEM medium containing 10% fetal bovine serum and 1% double-antibiotic) at 5% CO2 and 37°C. When the cell growth density reached 80%, the cells were trypsinized and resuspended. The resuspended cells were adjusted to the density with complete medium and inoculated into a 24-well plate at 0.6 x 105 per well. After 24 h of culture, transfection was performed.

[0246] Preparation of siRNA transfection mixture: siRNA was prepared into 20 μM, 0.4 μM, and 0.2 μM stock solutions using PBS buffer, and then 1.5 μl of siRNA stock solution was taken per well to dilute with 48.5 μl of Opti-MEM medium as 50 μl of siRNA working solution; 3 μl of Lipofectamine 2000 was taken per well, and 47 μl of Opti-MEM medium was added to dilute as 50 μl of Lipofectamine 2000 working solution. RNAiMAX transfection reagent was diluted in 47 μl Opti-MEM medium, incubated at room temperature for 5 minutes, and then 50 μl of the transfection reagent working solution was prepared; then 50 μl of siRNA working solution was mixed with 50 μl of transfection reagent working solution to prepare siRNA transfection mixture. The MOCK group was prepared by mixing 50 μl of Opti-MEM medium with 50 μl of transfection reagent working solution.

[0247] On the day of transfection, 500 μl of Opti-MEM medium was added to each well of a 24-well plate, and 100 μl of siRNA transfection mixture or MOCK transfection mixture was added to each well, so that the final concentration of siRNA was 50 nM / well, 1 nM / well, and 0.5 nM / well, respectively.

[0248] After 24 hours of continuous culture, total RNA was extracted according to the chemical extraction method. After removing the culture medium from each well, 1 mL of Trizol solution was added, and the cells were lysed by blowing. The lysate was transferred to a 1.5 mL centrifuge tube, 200 μL of chloroform was added, and the mixture was shaken well and incubated at room temperature for 3 min. Centrifugation was performed at 4°C and 12000 rpm for 10 min. 400 μL of supernatant was transferred to a centrifuge tube containing 400 μL of isopropanol, mixed well, and incubated at room temperature for 10 min. Centrifugation was performed at 4°C and 12000 rpm for 10 min, and the supernatant was discarded. 1 mL of 75% ethanol was added, and the centrifuge tube was inverted to wash the precipitate. Centrifugation was performed at 4°C and 12000 rpm for 5 min, the supernatant was removed, and the total RNA was obtained after air-drying at room temperature.

[0249] Detection of target gene mRNA expression level: 1 μg of the above total RNA was used for real-time fluorescent quantitative PCR according to the method described in the TaqMan Fast Advanced Master Mix (ABI Company, Catalog number: 4444557) instruction book. The reaction system was configured as follows: 20 μL of Real-time PCR reaction system, 5 μL of cDNA solution obtained by reverse transcription, 10 μL of TaqMan Fast Advanced Master Mix, 0.4 μL of 10 μM upstream primer, 0.4 μL of 10 μM downstream primer, 0.2 μL of 10 μM probe, and 3 μL of RNase-Free H2O. The prepared reaction system was placed in a real-time fluorescent quantitative PCR instrument (ABI Company, StepOnePlus TM / 7500), and two-step Real-time PCR amplification was performed. The amplification program was as follows: 50°C pre-denaturation for 2 min, 95°C pre-denaturation for 20 s, then 95°C denaturation for 3 s, 60°C annealing for 30 s, and repeating the denaturation and annealing process for 40 cycles.

[0250] Table 7: Primer sequence information used in Example 2:

[0251] The results of Example 2 (Tables 8-9) show that all of the designed sequences effectively reduced the expression of URAT1 mRNA, with RX591008, RX591017, RX591023, RX591047, RX591063, RX591066, RX591075, and RX591078 sequences having relatively better gene inhibition levels.

[0252] Table 8: Inhibition activity of siRNA sequences on target gene URAT1 in HuH1 cells in Example 2 at a final transfection concentration of 50 nM / well:

[0253] Table 9: Inhibition activity of siRNA sequences on target gene URAT1 in HuH1 cells in Example 2 at a final transfection concentration of 1 nM / well and 0.5 nM / well:

[0254] The above detailed description merely illustrates the content of the present application and does not represent a limitation thereof. Various modifications and improvements can be made by those of ordinary skill in the art without departing from the spirit and essential characteristics of the present disclosure, and these modifications and improvements are also considered to be within the scope of the present disclosure.

Claims

1. A double-stranded oligonucleotide that inhibits the expression of a URAT1 gene, characterized in that: The double-stranded oligonucleotide comprises a sense strand and an antisense strand, the length of the sense strand and the antisense strand is 17-30 nucleotides respectively, and the sense strand and the antisense strand are at least partially complementary; wherein, starting from the 5' end, the antisense strand comprises at least 17 consecutive nucleotides differing from no more than 3 nucleotides in the 1st-19th nucleotides of any one of the nucleotide sequences shown in SEQ ID NO. 70-138, 287-296.

2. The double-stranded oligonucleotide of claim 1, wherein: The sense strand comprises at least 17 consecutive nucleotides differing from no more than 3 nucleotides in any one of the nucleotide sequences shown in SEQ ID NO. 1-69, 277-286; Optionally, the length of the antisense strand is 21-23 nucleotides; the length of the sense strand is 19-21 nucleotides.

3. The double-stranded oligonucleotide of claim 1, wherein: Starting from the 5' end, the antisense strand comprises at least 17 consecutive nucleotides differing from no more than 3 nucleotides in the 1st-19th nucleotides of any one of the nucleotide sequences shown in SEQ ID NO. 70-138, 287-296; Optionally, starting from the 5' end, the antisense strand comprises at least 17 consecutive nucleotides differing from no more than 2 nucleotides in the 1st-19th nucleotides of any one of the nucleotide sequences shown in SEQ ID NO. 70-138, 287-296; Optionally, starting from the 5' end, the antisense strand comprises at least 17 consecutive nucleotides differing from no more than 1 nucleotide in the 1st-19th nucleotides of any one of the nucleotide sequences shown in SEQ ID NO. 70-138, 287-296; Optionally, starting from the 5' end, the antisense strand comprises at least 17 consecutive nucleotides selected from the 1st-19th nucleotides of any one of the nucleotide sequences shown in SEQ ID NO. 70-138, 287-296; Optionally, the antisense strand is selected from any one of the nucleotide sequences shown in SEQ ID NO. 70-138, 287-296.

4. The double stranded oligonucleotide according to any one of claims 1 to 3, characterized in that: The sense strand and the antisense strand have no more than 3 nucleotide mismatches; Optionally, the sense strand and the antisense strand have no more than 2 or 1 nucleotide mismatches; Optionally, the sense strand and the antisense strand are completely complementary.

5. The double-stranded oligonucleotide according to any one of claims 1 to 4, characterized in that: The double-stranded oligonucleotide comprises a sense strand sequence as set forth in any one of Set RN591001-RN591079, and a sequence of nucleotides 1-19, counting from the 5' terminus, of an antisense strand sequence as set forth in any one of Set RN591001-RN591079: Optionally, the double-stranded oligonucleotide is selected from any one of the groups RN591001-RN591079.

6. The double-stranded oligonucleotide of any one of claims 1-5, wherein: The double-stranded oligonucleotide comprises at least one modified nucleotide; Optionally, each nucleotide in the double-stranded oligonucleotide is selected from a modified nucleotide; Optionally, the modified nucleotide includes 2'-halogenated nucleotides, 2'-deoxygenated nucleotides, and 2'-OC nucleotides. 1-6 Alkyl-modified nucleotides, 2'-O-(CH2) n -O-Me modified nucleotides, 2'-amino modified nucleotides, 2',2'-[halogen, halogen] disubstituted or 2',2'-[halogen, C] disubstituted nucleotides 1-6 At least one of the following: alkyl] disubstituted nucleotides, debased nucleotides, or nucleotide-like molecules; n is selected from 1 or 2; the nucleotide-like molecules include at least one of peptide nucleic acid (PNA), morpholine nucleic acid (MNA), bridged nucleic acid (BNA), locked nucleic acid (LNA), glycol nucleic acid (GNA), threonucleotide (TNA), or non-locked nucleic acid (UNA); Optionally, the modified nucleotide comprises at least one of a 2'-F modified nucleotide, a 2'-deoxy modified nucleotide, a 2'-O-CH3 modified nucleotide, a 2'-O-CH2CH2-O-CH3 modified nucleotide, or a 2',2'-[F,CH3] double-substituted modified nucleotide.

7. The double stranded oligonucleotide of claim 6, wherein: The antisense strand of the double-stranded oligonucleotide is 21 nucleotides in length, the sense strand is 19 nucleotides in length, and each nucleotide in the double-stranded oligonucleotide is a modified nucleotide; wherein: at least four of positions 2, 6, 9, 14, 16 of the antisense strand are selected from 2'-F modified nucleotides, and the remaining positions are selected from 2'-0-CH3 modified nucleotides or 2'-0-CH2CH2-0-CH3 modified nucleotides; at least three of positions 7-10 of the sense strand are selected from 2'-F modified nucleotides, and the remaining positions are 2'-0-CH3 modified nucleotides; optionally, in the 5' to 3' direction, the antisense strand has 2'-F modified nucleotides at positions 2, 6, 9, 14, 16, and a 2'-0-CH2CH2-0-CH3 modified nucleotide at position 15, and 2'-0-CH3 modified nucleotides at the remaining positions; the sense strand has 2'-F modified nucleotides at positions 7-10, and 2'-0-CH3 modified nucleotides at the remaining positions; optionally, the double stranded oligonucleotide has at least one modified internucleotide linkage selected from a phosphorothioate internucleotide linkage, and the phosphorothioate internucleotide linkage is present at at least one of positions 1) - 5): 1) between the 1st and 2nd nucleotides, and / or between the 2nd and 3rd nucleotides, of the sense strand counting from the 5' terminus; 2) between the 1st and 2nd nucleotides, and / or between the 2nd and 3rd nucleotides, of the sense strand counting from the 3' terminus; 3) between the 1st and 2nd nucleotides, and / or between the 2nd and 3rd nucleotides, of the antisense strand counting from the 5' terminus; 4) between the 1st and 2nd nucleotides, and / or between the 2nd and 3rd nucleotides, and / or between the 3rd and 4th nucleotides, of the antisense strand counting from the 3' terminus; 5) between the 10th and 11th nucleotides of the antisense strand counting from the 5' terminus.

8. The double stranded oligonucleotide of claim 6, wherein: each nucleotide in the double stranded oligonucleotide is a modified nucleotide; wherein: in the 5' to 3' direction, at least four of positions 2, 6, 9, 12, 14, 16 of the antisense strand are selected from 2'-F modified nucleotides, and the remaining nucleotide positions other than the 2'-F modified nucleotides are each independently selected from 2'-0-CH3 modified nucleotides or 2'-0-CH2CH2-0-CH3 modified nucleotides; at least three of positions 7-10 of the sense strand are selected from 2'-F modified nucleotides, and the remaining nucleotide positions other than the 2'-F modified nucleotides are selected from 2'-0-CH3 modified nucleotides; Optionally, in the 5' end to 3' end direction, the 2nd, 6th, 14th, 16th nucleotides of the antisense strand are selected from 2'-F modified nucleotides, any one of the 9th and 12th nucleotides is selected from 2'-F modified nucleotides, another nucleotide is selected from 2'-O-CH3 modified nucleotides, the 15th nucleotide is selected from 2'-O-CH2CH2-O-CH3 modified nucleotides or 2'-O-CH3 modified nucleotides, and the remaining nucleotides are 2'-O-CH3 modified nucleotides; the 7th-10th nucleotides of the sense strand are selected from 2'-F modified nucleotides, and the remaining nucleotides are selected from 2'-O-CH3 modified nucleotides; Optionally, in the 5' end to 3' end direction, the 2nd, 6th, 12th, 14th, 16th nucleotides of the antisense strand are selected from 2'-F modified nucleotides, and the remaining nucleotides are 2'-O-CH3 modified nucleotides; the 7th-10th nucleotides of the sense strand are selected from 2'-F modified nucleotides, and the remaining nucleotides are selected from 2'-O-CH3 modified nucleotides; Optionally, the double-stranded oligonucleotide further has at least one modified internucleoside linkage, the modified internucleoside linkage comprises at least one of a phosphorothioate internucleoside linkage or a phosphorodithioate internucleoside linkage, and each of the at least one modified internucleoside linkage is independently selected from between the 1st and 2nd nucleotides of the sense strand starting from the 5' end, between the 2nd and 3rd nucleotides of the sense strand starting from the 5' end, between the 1st and 2nd nucleotides of the sense strand starting from the 3' end, between the 1st and 2nd nucleotides of the antisense strand starting from the 5' end, between the 2nd and 3rd nucleotides of the antisense strand starting from the 5' end, between the 10th and 11th nucleotides of the antisense strand starting from the 5' end, between the 1st and 2nd nucleotides of the antisense strand starting from the 3' end, or between the 2nd and 3rd nucleotides of the antisense strand starting from the 3' end. Optionally, the modified internucleoside linkage is selected from a phosphorothioate internucleoside linkage; Optionally, the 1stnucleotide of the antisense strand, counting from the 5' terminus, is selected from or a 5'-phosphate analog modified nucleotide; the 5'-phosphate analog modified nucleotide is selected from any one of the nucleotides shown in the following structure: wherein Base represents a nucleobase selected from A, U, G, C or T; R represents -H, -OH, -CH3, -OCH3, -F; Optionally, the 5'-phosphate analog modified nucleotide is selected from a 5'-(E)-vinyl phosphonate (5'-(E)-VP) modified nucleotide; Optionally, the 1stnucleotide of the antisense strand, counting from the 5' terminus, is selected from or the 1st nucleotide of the antisense strand starting from the 5' end is selected from a 5'-(E)-VP modified nucleotide.

9. The double-stranded oligonucleotide according to any one of claims 6-7, wherein: the antisense strand comprises at least 17 contiguous nucleotides differing by no more than 3 nucleotides from nucleotides 1-19 of the nucleotide sequence recited in any one of SEQ ID NOs. 208-276, 307-316; and the sense strand comprises at least 17 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence recited in any one of SEQ ID NOs. 139-207, 297-306; optionally, the antisense strand comprises at least 17 contiguous nucleotides differing by no more than 3 nucleotides from nucleotides 1-19 of the nucleotide sequence recited in any one of SEQ ID NOs. 208-276, 307-316; optionally, the antisense strand comprises at least 17 contiguous nucleotides differing by no more than 2 nucleotides from nucleotides 1-19 of the nucleotide sequence recited in any one of SEQ ID NOs. 208-276, 307-316; optionally, the antisense strand comprises at least 17 contiguous nucleotides differing by no more than 1 nucleotide from nucleotides 1-19 of the nucleotide sequence recited in any one of SEQ ID NOs. 208-276, 307-316; optionally, the antisense strand comprises at least 17 contiguous nucleotides differing by no more than 1 nucleotide from nucleotides 1-19 of the nucleotide sequence recited in any one of SEQ ID NOs. 208-276, 307-316; optionally, the antisense strand comprises at least 17 contiguous nucleotides differing by no more than 1 nucleotide from nucleotides 1-19 of the nucleotide sequence recited in any one of SEQ ID NOs. 208-276, 307-316; Optionally, the double-stranded oligonucleotide is selected from the sense strand sequences set forth in any one of Groups RX591001-RX591079, and the 1st-19th nucleotide sequences, counting from the 5' terminus, of the antisense strand sequences set forth in any one of Groups RX591001-RX591079; optionally, the antisense strand is selected from the nucleotide sequence recited in any one of SEQ ID NOs. 208-276, 307-316; 10. Conjugate, characterized in that: optionally, the double stranded oligonucleotide is selected from any one of the groups RX591001-RX591079. comprising the double stranded oligonucleotide of any one of claims 1-9 and one or more targeting ligand groups conjugated to the double stranded oligonucleotide; optionally, the targeting ligand group targets the kidney Megalin receptor or the kidney natriuretic peptide receptor; 1) -X1(KKEEE) n - K m - wherein n = 1-5, m = 0 or 1 or 2, X1is selected from any L-alpha amino acid or L-beta amino acid except K, E); 2) - CLPVASC, preferably in the form of a cyclic peptide 3) -CYFQNC-, preferably in the form of a cyclic peptide optionally, the targeting ligand group is selected from a polypeptide fragment comprising at least 4 amino acids, and the targeting ligand group comprises any one of the following 1)-6) polypeptide fragments: 4) -KIDRI-; 5) -IDRI- (Ile-Asp-Arg-Ile-); 6) -dXa-Ser-dXb-X2-dXc-Gly-Xd-Ile-Asp-Arg(Ak)-Ile-; wherein X2 is selected from any unnatural amino acid; dXa, dXb, dXc are selected from any D-amino acid; Arg(Ak) is selected from arginine or alkylated modified arginine; Xd is selected from Hyp or Pro; preferably, the targeting ligand group is selected from a polypeptide comprising any one of the following a)-g) amino acid sequence segments: a) -KKEEE-KKEEE-KKEEE-K-; b) -CKKEEE-KKEEE-KKEEE-K; c) -KKEEE-KKEEE-K-; d) -KIDRI-; e) -DPhe-Ser-DHyp-Cha-DAla-Gly-Hyp-Ile-Asp-Arg(Me)-Ile-; f) -DPhe-Ser-DHyp-Cha-DAla-Gly-Hyp-Ile-Asp-Arg(Ile)-Ile-; g) -DPhe-Ser-DHyp-Cha-DAla-Gly-Hyp-Ile-Asp-Arg-Val-; wherein Cha is selected from D-Cha or L-Cha; and wherein Me is selected from methyl or hydrogen. c) -CLPVASC-, preferably in the form of a cyclic peptide ​ e) -CYFQNCPRG-, preferably in the form of a cyclic peptide ​ g) -DPhe-Ser-DPro-Cha-DAla-Gly-Pro-Ile-Asp-Arg-Ile-.

11. The conjugate of claim 10, wherein, the targeting ligand group is linked to the double stranded oligonucleotide via a linker; Optionally, the structural unit of the targeting ligand group-linker in the conjugate is selected from any of the structures shown in 1) - 7) below: 1) La-KKEEE-KKEEE-KKEEE-K-Laa; 2) La-CKKEEE-KKEEE-KKEEE-K-Laa; 3) La-CLPVASC-Laa, preferably a cyclic peptide 4) La-DPhe-Ser-DHyp-Cha-DAla-Gly-Hyp-Ile-Asp-Arg(Me)-Ile-Laa; 5) La-CYFQNCPRG-Laa; preferably in cyclic peptide form 6) La-DPhe-Ser-DPro-Cha-DAla-Gly-Pro-Ile-Asp-Arg-Ile-Laa; 7) Lb-KIDRI-Laa; In formulae 1) - 6), either La or Laais a linker, and the other is a terminal blocking group; the linker is a linking group comprising at least one of a triazole group or a PEG unit or an acyl group; Optionally, La is a linker, and the linker structure comprises -PEG2-CH2CH2CO- or -triazole-PEG2-; In formula 7), Lb is a linking group comprising a 6-10 membered aromatic or heteroaromatic ring, and Laais independently selected from an amino group, or an alkyl-substituted amine group, such as -NHCH3.

12. The conjugate of claim 11, characterized in that: the targeting ligand group is linked to the double stranded oligonucleotide via a linker; 13. The conjugate according to claim 11 or 12, characterized in that: the structural unit of the targeting ligand group-linker in the conjugate is selected from any of the structures shown in a) - j), or a pharmaceutically acceptable salt thereof: a) -La-KKEEE-KKEEE-KKEEE-K-Lc; b) -La-CKKEEE-KKEEE-KKEEE-K-Lc; c) -La-CLPVASC-Lc; preferably in the form of a cyclic peptide d) -La-DPhe-Ser-DHyp-Cha-DAla-Gly-Hyp-Ile-Asp-Arg(Me)-Ile-Lc; e) -La-CYFQNCPRG-Lc; preferably a cyclic peptide f) -La-DPhe-Ser-DPro-Cha-DAla-Gly-Pro-Ile-Asp-Arg-Ile-Lc; g) h) i) j) wherein La is a linker, which is independently selected from any bond, -NH-, an amide group, or a linking group comprising at least one PEG unit; Optionally, La is a linking group comprising at least 2 PEG units; Optionally, La is selected from an amido group, or n is 1-3, m is 0 or 1, and Z is -CO- (carbonyl) or -NH-; wherein Lcis a terminal blocking group; optionally, it is a C-terminal blocking group, and Lcis selected from an amino group, or an alkyl-substituted amine group, preferably -NHCH3.

14. A pharmaceutical composition, characterized by: The pharmaceutical composition comprises the double stranded oligonucleotide of any one of claims 1-9, and / or the conjugate of any one of claims 10-13.

15. Use of the double-stranded oligonucleotide of any one of claims 1-9, and / or of the conjugate of any one of claims 10-13, and / or of the pharmaceutical composition of claim 14, for the manufacture of a medicament for inhibiting the expression of the URAT1 gene.

16. Use of the double-stranded oligonucleotide of any one of claims 1-9, and / or of the conjugate of any one of claims 10-13, and / or of the pharmaceutical composition of claim 14, for the manufacture of a medicament for the treatment and / or prevention of a pathological condition or disease associated with the expression of URAT1.

17. Use according to claim 16, characterized in that: The pathological condition or disease is hyperuricemia or gout.

18. A method of inhibiting the expression of a URAT1 gene, characterized in that, It comprises administering to a subject an effective dose of the double-stranded oligonucleotide of any one of claims 1-9, and / or of the conjugate of any one of claims 10-13, and / or of the pharmaceutical composition of claim 14; Optionally, the subject is a human.

19. A method for the prophylaxis and / or treatment of hyperuricemia or gout, characterized by: The method comprises administering to a subject an effective dose of the double-stranded oligonucleotide of any one of claims 1-9, and / or of the conjugate of any one of claims 10-13, and / or of the pharmaceutical composition of claim 14.