WO2025201416A1 - ACIDE NUCLÉIQUE CIBLANT LA SOUS-UNITÉ βE DE L'INHIBINE ET UTILISATION ASSOCIÉE - Google Patents

ACIDE NUCLÉIQUE CIBLANT LA SOUS-UNITÉ βE DE L'INHIBINE ET UTILISATION ASSOCIÉE

Info

Publication number
WO2025201416A1
WO2025201416A1 PCT/CN2025/085076 CN2025085076W WO2025201416A1 WO 2025201416 A1 WO2025201416 A1 WO 2025201416A1 CN 2025085076 W CN2025085076 W CN 2025085076W WO 2025201416 A1 WO2025201416 A1 WO 2025201416A1
Authority
WO
WIPO (PCT)
Prior art keywords
nucleotides
nucleic acid
seq
nucleotide
sequence shown
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/CN2025/085076
Other languages
English (en)
Chinese (zh)
Inventor
郁东
蒋伟文
兰涛
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Synerk Biopharmaceuticals Ltd
Original Assignee
Synerk Biopharmaceuticals Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Synerk Biopharmaceuticals Ltd filed Critical Synerk Biopharmaceuticals Ltd
Publication of WO2025201416A1 publication Critical patent/WO2025201416A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1136—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against growth factors, growth regulators, cytokines, lymphokines or hormones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/549—Sugars, nucleosides, nucleotides or nucleic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0684—Cells of the urinary tract or kidneys
    • C12N5/0686—Kidney cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/14—Type of nucleic acid interfering nucleic acids [NA]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2510/00—Genetically modified cells

Definitions

  • the present invention relates to the field of biotechnology, in particular to a nucleic acid targeting inhibin subunit ⁇ E and uses thereof.
  • Obesity is closely linked to a variety of chronic diseases. Currently, over 500 million people worldwide are overweight or obese. Obesity is thought to increase the risk of cardiovascular disease, hypertension, diabetes, gout, and other diseases (Yusuf, S. et al. Obesity and the risk of myocardial infarction in 27,000 participants from 52 countries: a case-control study. Lancet 366, 1640–1649 (2005).). Although the mechanisms of fat distribution are not yet fully understood, abdominal fat accumulation is associated with cardiovascular and metabolic diseases. Increased waist-to-hip ratio, adjusted for body mass, has been associated with type 2 diabetes and cardiovascular disease, as well as other lipid and blood pressure abnormalities (Emdin, C.A. et al.
  • the purpose of the present invention is to overcome the problems existing in the prior art and provide a new nucleic acid targeting INH ⁇ E and its use.
  • the first aspect of the present invention provides a nucleic acid, which includes a sense chain and an antisense chain, wherein the sense chain contains at least 15 consecutive nucleotides, and the at least 15 consecutive nucleotides differ from any at least 15 consecutive nucleotides in the sequence shown in any one of SEQ ID No. 1 to 68 by no more than 3 nucleotides; or the antisense chain contains at least 15 consecutive nucleotides, and the at least 15 consecutive nucleotides differ from any at least 15 consecutive nucleotides in the nucleotide sequence shown in any one of SEQ ID No. 69 to 136 by no more than 3 nucleotides.
  • a second aspect of the present invention provides a targeted drug delivery system, which comprises a targeting group, a linking group, and the nucleic acid as described above connected to the targeting group via the linking group.
  • the third aspect of the present invention provides an in vitro cell containing the nucleic acid.
  • the fourth aspect of the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising the nucleic acid or targeted drug delivery system as described above and a pharmaceutically acceptable carrier.
  • the fifth aspect of the present invention provides a method for inhibiting the expression of inhibin subunit ⁇ E in a cell, the method comprising: contacting the cell with the nucleic acid, the targeted drug delivery system or the pharmaceutical composition to inhibit the expression of inhibin subunit ⁇ E in the cell.
  • FIG1 shows the results of the inhibition of INHBE gene expression by different targeted drug delivery systems in Example 2 of the present invention.
  • nucleic acid refers to a composition containing RNA or RNA-like (e.g., chemically modified RNA) oligonucleotide molecules that can degrade or inhibit (e.g., degrade or inhibit under appropriate conditions) the translation of messenger RNA (mRNA) transcripts of target mRNA in a sequence-specific manner.
  • the nucleic acid can act through an RNA interference mechanism (i.e., by inducing RNA interference through interaction with the RNA interference pathway mechanism of mammalian cells (RNA-induced silencing complex or RISC)), or through any alternative mechanism or pathway.
  • RNA interference mechanism i.e., by inducing RNA interference through interaction with the RNA interference pathway mechanism of mammalian cells (RNA-induced silencing complex or RISC)
  • nucleic acids disclosed herein includes, but is not limited to, short (or small) interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), short hairpin RNA (shRNA), and dicer substrates.
  • siRNA short (or small) interfering RNA
  • dsRNA double-stranded RNA
  • miRNA microRNA
  • shRNA short hairpin RNA
  • the antisense strand contains at least 15 consecutive nucleotides, and the at least n consecutive nucleotides differ by no more than 3 nucleotides compared with any at least 15 consecutive nucleotides in the reference sequence (such as the sequence shown in SEQ ID No. X or the nucleotide sequence at positions 1 to 21 of the shown sequence)
  • the comparison situation involved includes the comparison of at least 15 consecutive nucleotides starting from any position in the reference sequence (such as position 1, 2, 3, ..., 7, or 8).
  • the antisense strand contains 21 consecutive nucleotides, wherein the nucleotide sequence at positions 1 to 15 has 1, 2 or 3 differences with the nucleotide sequence at positions 2 to 16 of the reference sequence, and the nucleotides at positions 16 to 20 in the antisense strand (sense strand) are the same as or different from the nucleotide sequence at positions 17 to 21 of the reference sequence.
  • sequences are also within the scope of the present invention.
  • the antisense strand contains 21 consecutive nucleotides, wherein the nucleotide sequences at positions 1 to 15 are identical to the nucleotide sequences at positions 1 to 15 of the reference sequence, and the nucleotide sequences at positions 16 to 21 of the antisense strand (sense strand) are identical to or different from the nucleotide sequences at positions 16 to 21 of the reference sequence.
  • sequences are also within the scope of the present invention.
  • the “difference” or “difference” mentioned includes one or more of substitution, insertion, and deletion.
  • INH ⁇ E inhibin ⁇ E
  • TGF- ⁇ transforming growth factor ⁇
  • INH ⁇ E plays a role in regulating various cellular processes, including cell proliferation, apoptosis, immune responses, and hormone secretion.
  • the terms “silencing,” “reducing,” “inhibiting,” “downregulating,” or “knockdown” when referring to the expression of a given gene mean that the expression of the gene is reduced when the cell, cell population, tissue, organ, or subject is treated with a nucleic acid described herein, as measured by the level of RNA transcribed from the gene or the level of a polypeptide, protein, or protein subunit translated from mRNA in the cell, cell population, tissue, organ, or subject in which the gene is transcribed, compared to a second cell, cell population, tissue, organ, or subject that has not been so treated.
  • nucleobase or nucleotide sequence molecules As used herein, “fully complementary” means that in a hybridization pair of nucleobase or nucleotide sequence molecules, all (100%) bases in the contiguous sequence of the first oligonucleotide hybridize to the same number of bases in the contiguous sequence of the second oligonucleotide.
  • the contiguous sequence may comprise all or part of the first nucleotide sequence or the second nucleotide sequence.
  • partially complementary means that in a hybridization pair of nucleobase or nucleotide sequence molecules, at least 70%, but not all, of the bases in the contiguous sequence of the first oligonucleotide hybridize to the same number of bases in the contiguous sequence of the second oligonucleotide.
  • the contiguous sequence may comprise all or part of the first nucleotide sequence or the second nucleotide sequence.
  • substantially complementary means that in a hybridization pair of nucleobase or nucleotide sequence molecules, at least 85%, but not all, of the bases in the contiguous sequence of the first oligonucleotide hybridize to the same number of bases in the contiguous sequence of the second oligonucleotide.
  • the contiguous sequence may comprise all or part of the first nucleotide sequence or the second nucleotide sequence.
  • the term "at least partially complementary” means that the first oligonucleotide and the second oligonucleotide are partially complementary, substantially complementary or completely complementary in a hybridization pair of nucleobase or nucleotide sequence molecules.
  • treating refers to methods or steps taken to provide relief or alleviation of the number, severity, and/or frequency of one or more disease symptoms in a subject.
  • Such treatment may include prevention, management, prophylactic treatment, and/or inhibition or reduction of the number, severity, and/or frequency of one or more disease symptoms in a subject.
  • connection means that two compounds or molecules are joined by a covalent bond. Unless otherwise indicated, as used herein, the term “connection” may refer to a connection between a first compound and a second compound with or without any intermediate atoms or groups of atoms.
  • the sense strand has 15 to 30 (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides (bases).
  • the antisense strand comprises at least 15, 16, 17, 18, 19, 20, 21, 22, or 23 consecutive nucleotides, and the at least 15, 16, 17, 18, 19, 20, 21, 22, or 23 consecutive nucleotides differ by no more than 0, 1, 2, or 3 nucleotides compared to at least 15, 16, 17, 18, 19, 20, 21, 22, or 23 consecutive nucleotides in any of the sequences set forth in SEQ ID No. 115 or 120; and the sense strand comprises a nucleotide sequence that is at least partially complementary (e.g., partially complementary, substantially complementary, or completely complementary) to the antisense strand.
  • the antisense strand has the above sequence, the double-stranded RNA has a significantly better inhibitory effect on INH ⁇ E.
  • the positive strand contains a nucleotide sequence that differs from the sequence shown in SEQ ID NO: 52 (5’-GCCUGGCUUAUACUUUCUUAA-3’) by 0, 1 or 2 nucleotides
  • the antisense strand contains a nucleotide sequence that differs from the sequence shown in SEQ ID NO: 120 (5’-UUAAGAAAGUAUAAGCCAGGCUU-3’) by 0, 1 or 2 nucleotides.
  • the positive strand contains a nucleotide sequence that differs from the sequence shown in SEQ ID NO: 139 (5’-CGAGACAAGCAUUUAUACUUG-3’) by 0, 1 or 2 nucleotides
  • the antisense strand contains a nucleotide sequence that differs from the sequence shown in SEQ ID NO: 140 (5’-CAAGUAUAAAUGCUUGUCUCGUU-3’) by 0, 1 or 2 nucleotides.
  • nucleotide groups in the above nucleic acid may be unmodified or may contain at least one modified nucleotide group, and the modification may be at any nucleotide position.
  • the sense and antisense strands can be partially complementary, substantially complementary, or fully complementary to each other.
  • the sequence identity of the sense strand or antisense strand in the nucleic acid to the corresponding sequence mentioned in the present invention is less than 100% or differs by more than 1 nucleotide, it still has an inhibitory effect on INH ⁇ E that is similar to the corresponding sequence (such as still having an efficacy equivalent to 80-120%, 85-115% or 90-110% of the corresponding sequence) or equivalent (such as still having an efficacy equivalent to 95-105% of the corresponding sequence).
  • the two bases at the 3' end of the antisense strand (such as the sequence shown in any one of SEQ ID No. 69 to 136) are replaced with AA, CU, UC, AG, CC, GG or UG, etc., or a combination of any two nucleic acids.
  • Such nucleic acid sequences also fall within the scope of protection of the present invention.
  • column 1 indicates the position of the first base of the targeted gene in the INH ⁇ E coding sequence, and so on.
  • the numbers in columns 3 and 5 indicate the sequence number, for example, "1" indicates SEQ ID No. 1.
  • the reference sequence of the targeted gene is the human INH ⁇ E coding sequence NM_031479.
  • the nucleic acid containing the modifying group has an inhibition efficiency of INH ⁇ E of not less than 50% (e.g., 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%).
  • 50% e.g., 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%
  • modification of the phosphate group refers to modification of the oxygen in the phosphate group, including phosphorothioate and boranophosphate modifications. As shown in the following formulas, the oxygen in the phosphate group is replaced with sulfur, borane, amine, alkyl, or alkoxy groups. These modifications can stabilize the structure of nucleic acids and maintain high base pairing specificity and affinity.
  • BASE represents the base A, U, C, G, or T.
  • X can be oxygen (O) or sulfur (S).
  • R in the above structure can be the same or different, for example: hydrogen (H), fluorine (F), methoxy (OME) or methoxyethyl (MOE), hydroxyl, allyl, ethylamino, propargyl, amino, cyanoethyl, acetyl, etc.
  • R' and R" can each independently be hydrogen (H), methyl (CH3), ethyl (CH2CH3), propyl (CH2CH2CH3), isopropyl (CH(CH3)2), allyl, propargyl, acyloxybenzyl, or acyloxyethyl.
  • modification of the ribose group refers to modification of the 2′-hydroxyl group (2′-OH) in the ribose group.
  • Introducing certain substituents, such as methoxy or fluorine, at the 2′-hydroxyl position of the ribose group makes the nucleic acid less susceptible to cleavage by ribonucleases, thereby increasing the stability of the nucleic acid and making it more resistant to nuclease hydrolysis.
  • Modifications of the 2′-hydroxyl group in nucleotide pentoses include 2′-fluoro modification (such as 2′-arabino-fluoro modification), 2′-methoxy modification (2′-OME), 2′-methoxyethyl modification (2′-MOE), 2′-2,4-dinitrophenol modification (2′-DNP modification), 2′,4′-constrained ethyl modification, 2′-amino modification, 2′-deoxy modification, BNA, acyclic nucleic acid modification, misaligned nucleic acid modification, and L-type nucleic acid modification.
  • BNA endocyclic bridged nucleotide refers to a constrained or inaccessible nucleotide.
  • Acyclic nucleic acids are nucleotides formed by opening the sugar ring of the nucleotide, such as unlocked nucleic acid (UNA) nucleotides and glycerol nucleic acid (GNA) nucleotides.
  • Unlocked nucleic acid (UNA) nucleotides and glycerol nucleic acid (GNA) nucleotides.
  • Misplaced nucleic acid modification refers to the replacement of a 3', 5'-phosphate bond link by a 2', 5'-phosphate bond chain.
  • L-type nucleic acid modification refers to the replacement of a naturally occurring D-type nucleic acid with its mirror image stereo equivalent, an L-type nucleic acid.
  • BASE represents the base A, U, C, G or T.
  • R in the above structure can be the same or different, such as hydrogen (H), fluorine (F), methoxy (OME) or methoxyethyl (MOE), hydroxyl, allyl, ethylamino, propargyl, cyanoethyl, acetyl, etc.
  • the nucleotide group containing a uracil base or a cytosine base in the sense strand of the RNAi agent is a nucleotide group in which the ribose group is modified, that is, the 2'-OH group of the ribose group in the nucleotide group containing a uracil base or a cytosine base in the sense strand of the RNAi agent is replaced by a methoxy group or a fluorine group.
  • the 3' end of both the sense strand and the antisense strand of the RNAi agent can be connected to dTdT; or, the 3' end of the antisense strand of the RNAi agent can be connected to AA or UU or a combination of any two nucleic acids (which can be but is not limited to CC, GG or UG), so that the sequence has a specific inducement for mRNA degradation.
  • the RNAi agent with the above modifications exhibits a more excellent in vivo inhibitory effect, and the above modifications can further reduce the immunogenicity of the RNAi agent of the present invention in vivo.
  • the RNAi reagents of the present invention may also include a modification that includes a nucleoside monophosphate attached to the 5' end of the antisense strand.
  • the 5'-monophosphate at the terminal end of the siRNA guide strand is important for RISC recognition. Phosphorylation of the 5'-hydroxyl group plays a role in the effective loading of siRNA onto Ago2 within cells.
  • the monophosphate at the 5' end of the siRNA guide strand interacts with Argonaute-2 (Ago2) through H-bonding, ensuring accurate targeting and precise cleavage of the mRNA target.
  • Commonly used 5'-monophosphate nucleoside derivatives include the following.
  • phosphate nucleoside derivatives have been shown to exhibit a certain degree of stability in biological metabolic media and to be effective in promoting the loading of siRNA guide strands onto Ago2 within cells (Nucleic Acids Research, 2015, 43, 2993–3011).
  • trans-vinyl phosphate (VP) is preferably used as the primary choice, but monophosphate nucleoside derivatives other than those mentioned above may also be included.
  • BASE represents the base A, U, C, G, or T.
  • R in the above structures can be the same or different, such as hydrogen (H), fluorine (F), methoxy (OME) or methoxyethyl (MOE), hydroxyl, allyl, ethylamino, propargyl, cyanoethyl, amino, acetyl, etc.
  • the modified nucleotides are preferably selected from one or more of 2'-O-methyl nucleotides, 2'-fluoro nucleotides, 2'-deoxy nucleotides, 2',3'-open ring nucleotide mimics, locked nucleotides, 2'-F-arabino nucleotides, 2'-methoxyethyl nucleotides, abasic nucleotides, ribitols, inverted nucleotides, inverted 2'-O-methyl nucleotides, inverted 2'-deoxy nucleotides, 2'-amino modified nucleotides, 2'-alkyl modified nucleotides, morpholino nucleotides (Morpholino), peptide nucleic acids (PNA), glycerol nucleic acids (GNA), triazacyclic DNA (tcDNA), nucleotides containing non-natural bases, nucleot
  • the modified inter-linkage is preferably selected from one or more of phosphorothioate inter-nucleotide linkages and methylphosphonate inter-nucleotide linkages. In some embodiments, the modified inter-linkage is further preferably selected from one or more of phosphorothioate monoester inter-nucleotide linkages and phosphorothioate diester inter-nucleotide linkages.
  • the antisense strand comprises a 2'-fluoro nucleotide at nucleotide position 14 and at least one of nucleotide positions 1, 2, 3, 4, 6, 7, 8, 12, or 16 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, or 9) of the nucleotide sequence of any one of SEQ ID Nos. 69 to 136.
  • the above antisense strand modification scheme further enhances the inhibitory effect of the nucleic acid on INH ⁇ E.
  • the antisense chain has 2'-fluoro nucleotides at the 2nd, 6th, 14th and 16th nucleotide positions and at least one of the 1st, 3rd, 4th, 7th, 8th and 12th nucleotide positions (such as 1st, 2nd, 3rd, 4th, 5th or 6th nucleotide positions) of the nucleotide sequence shown in any one of SEQ ID No. 69 to 136, and the other positions have 2'-O-methyl nucleotides.
  • the sense strand comprises a 2'-fluoro nucleotide at nucleotide position 9 and at least one of nucleotide positions 1, 7, 10, 11, and 12 (e.g., 1, 2, 3, 4, or 5) of the nucleotide sequence of any one of SEQ ID Nos. 1-68.
  • the above sense strand modification scheme further enhances the inhibitory effect of the nucleic acid on INH ⁇ E.
  • the positive chain has 2'-fluoro nucleotides at the 7th, 9th and 11th nucleotide positions and at least one of the 1st, 10th and 12th nucleotide positions (such as 1st, 2nd or 3rd) of the nucleotide sequence shown in any one of SEQ ID No. 1 to 68, and the other positions have 2'-O-methyl nucleotides.
  • the last 2 to 4 (e.g., 2, 3, or 4) nucleotides at the 5' end and/or 3' end of the antisense strand contain a phosphorothioate internucleotide bond
  • the last 2 to 4 (e.g., 2, 3, or 4) nucleotides at the 5' end and/or 3' end of the sense strand contain a phosphorothioate internucleotide bond
  • a nucleotide represented by a lowercase letter indicates that the nucleotide is a 2'-O-methyl nucleotide; f indicates that the nucleotide adjacent to the left is a 2'-fluoro nucleotide; s indicates that the two adjacent nucleotides on the left and right are connected by a phosphorothioate diester internucleotide bond.
  • nucleic acids described herein can be obtained by conventional methods in the art, such as solid-phase synthesis and liquid-phase synthesis. Solid-phase synthesis is commercially available on a custom-built basis and can therefore be purchased commercially.
  • the modified nucleotide groups can be introduced by correspondingly modified nucleotide monomers.
  • nucleic acid (siRNA) of the present invention when applied to different targeted drug delivery systems, all has a better inhibitory effect.
  • the effect advantage of the naked sequence and the modified sequence in the present invention does not depend on the selection of the targeting vector.
  • the present invention has also optimized the targeted drug delivery system and obtained the following technical scheme.
  • the targeting group can further improve the targeting of small nucleic acids and can be provided by monosaccharides (such as glucose, mannose, allose, altrose, galactose, galactosamine, N-acetylgalactosamine, talose, fructose, idose, etc.) and/or polypeptides (such as proteins, monoclonal antibodies, nanobodies).
  • monosaccharides such as glucose, mannose, allose, altrose, galactose, galactosamine, N-acetylgalactosamine, talose, fructose, idose, etc.
  • polypeptides such as proteins, monoclonal antibodies, nanobodies.
  • the linking group may be selected from -O-[CH2CH2O]n-, -[CH2]m-CONH-[CH2]nO-, -O-[CH2CH2O]m-CONH-[CH2]nO-, and -O-[CH2]m-CONH-[CH2H2O]nO-, wherein m and n may each independently be an integer from 1 to 10.
  • the targeted drug delivery system comprises a ligand and the nucleic acid linked to the ligand, wherein the ligand is linked to one or more of the 5' end of the antisense strand, the 3' end of the antisense strand, the 5' end of the sense strand, and the 3' end of the sense strand.
  • the ligand is a GalNAc derivative.
  • the ligand is one or more GalNAc derivatives connected by single-stranded, double-stranded or triple-stranded branched linkers.
  • the RNAi agent comprises a compound having a structure shown in the following formula I:
  • Nu represents the duplex. This targeted drug delivery system utilizes the structural characteristics on the left side to improve the cell penetration ability of the nucleic acid drug (Nu) and enhance its intracellular stability. It also has a simple preparation process and strong practicality.
  • the present invention also provides an in vitro cell containing the nucleic acid.
  • the cells will not develop into an animal individual.
  • the cells can be microbial cells or animal cells, but the animal cells are not animal embryonic stem cells and cells in various formation and development stages (e.g., germ cells, fertilized egg cells, etc.).
  • inhibiting expression of inhibin subunit ⁇ E reduces the level of inhibin subunit ⁇ E protein in a serum sample of the subject by at least 50%, 60%, 70%, 80%, 90%, or 95%, e.g., compared to the level of inhibin subunit ⁇ E expression before the cell is first contacted with the RNAi agent or the pharmaceutical composition.
  • the present invention also provides the use of the nucleic acid, the targeted drug delivery system, or the pharmaceutical composition in treating and/or preventing diseases associated with the inhibin subunit ⁇ E.
  • a method for treating and/or preventing diseases associated with the inhibin subunit ⁇ E comprises administering the nucleic acid, the targeted drug delivery system, or the pharmaceutical composition to a subject.
  • the disease is selected from diseases related to abnormal fat distribution.
  • the disease is selected from at least one of cardiovascular disease, diabetes, lipid metabolism disorder, hypertension, obesity, and metabolic syndrome.
  • the subject can be a mammal, including a primate (such as a human, a non-human primate, such as a monkey and a chimpanzee), a non-primate (such as a cow, a pig, a horse, a goat, a rabbit, a sheep, a hamster, a guinea pig, a cat, a dog, a rat, or a mouse), or a bird.
  • a primate such as a human, a non-human primate, such as a monkey and a chimpanzee
  • a non-primate such as a cow, a pig, a horse, a goat, a rabbit, a sheep, a hamster, a guinea pig, a cat, a dog, a rat, or a mouse
  • a bird preferably a primate, more preferably a human.
  • the drug can be administered via a variety of routes, depending on whether local or systemic treatment is desired.
  • the dosage can be referred to above and will not be repeated here.
  • administration can be topical (e.g., transdermal patch), pulmonary, e.g., via inhalation or insufflation of a powder or spray, including via a nebulizer; intratracheal, nasal, epidermal, and transdermal, oral, or parenteral.
  • Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion; subcutaneous, e.g., via an implant device; or intracranial, e.g., via intraparenchymal, intrathecal, or intraventricular administration.
  • the nucleic acid, the targeted drug delivery system, or the pharmaceutical composition is administered to the subject by subcutaneous administration, intravenous administration, and/or intramuscular administration.
  • the measured parameters of raw material components may have slight deviations within the range of weighing accuracy unless otherwise specified.
  • acceptable deviations caused by instrument testing accuracy or operational accuracy are allowed.
  • HEK293T cells were passaged for 2-3 generations in DMEM (Gibco, 11995065) containing 10% FBS (Hyclone, SH30406.05), and then plated on 0.1% gelatin-coated 96-well plates (Greiner #655098) to a cell density of 30K per well and cultured overnight. The next day, siRNA samples were dissolved and diluted in ddH2O .
  • siRNA containing the duplex sequence of the present invention has different degrees of inhibitory effect on the expression of INH ⁇ E.
  • Each of the targeted drug delivery systems conjugated with TriGalNAc (see Formula I for its structure) listed in Table 5 was dissolved in 100 ⁇ L of enzyme-free sterile water to a 1000 ⁇ M solution, corresponding to a 1000 nM working solution.
  • Monkey primary hepatocytes were removed from liquid nitrogen, thawed and revived at 37°C, rinsed, counted, and centrifuged using PMonH plating medium. After removing the supernatant, the cells were diluted to 250 kb/mL using fresh PMonH plating medium. 100 ⁇ L of this diluted cell solution was plated onto a 96-well cell culture plate, with 25 kb cells per well.
  • siRNAs listed in Table 6 were serially diluted with enzyme-free sterile water and assayed in HEK293T cells using the Reneilla luciferase assay, as described in Example 1. IC50 values were calculated using Prism GraphPad, as shown in Table 7.
  • siRNAs listed in Table 8 were serially diluted with enzyme-free sterile water and assayed in HEK293T cells using the Reneilla luciferase assay method described in Example 1. The results are shown in Table 9.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Biomedical Technology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Diabetes (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Hematology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Obesity (AREA)
  • General Engineering & Computer Science (AREA)
  • Epidemiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Endocrinology (AREA)
  • Cardiology (AREA)
  • Urology & Nephrology (AREA)
  • Microbiology (AREA)
  • Emergency Medicine (AREA)
  • Cell Biology (AREA)
  • Child & Adolescent Psychology (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne un acide nucléique ciblant la sous-unité βE de l'inhibine et une utilisation associée. L'acide nucléique peut inhiber efficacement l'expression INHβE et peut, ainsi, traiter et/ou prévenir des maladies associées à une expression accrue d'INHβE.
PCT/CN2025/085076 2024-03-27 2025-03-26 ACIDE NUCLÉIQUE CIBLANT LA SOUS-UNITÉ βE DE L'INHIBINE ET UTILISATION ASSOCIÉE Pending WO2025201416A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202410359219.4 2024-03-27
CN202410359219 2024-03-27

Publications (1)

Publication Number Publication Date
WO2025201416A1 true WO2025201416A1 (fr) 2025-10-02

Family

ID=96102166

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2025/085076 Pending WO2025201416A1 (fr) 2024-03-27 2025-03-26 ACIDE NUCLÉIQUE CIBLANT LA SOUS-UNITÉ βE DE L'INHIBINE ET UTILISATION ASSOCIÉE

Country Status (2)

Country Link
CN (1) CN120210203A (fr)
WO (1) WO2025201416A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN120210203A (zh) * 2024-03-27 2025-06-27 施能康医药科技(苏州)有限公司 靶向抑制素亚单位βE的核酸及其用途

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116583291A (zh) * 2020-12-14 2023-08-11 雷杰纳荣制药公司 用抑制素亚基βE(INHBE)抑制剂治疗代谢病症和心血管疾病的方法
CN117716032A (zh) * 2021-07-21 2024-03-15 阿尔尼拉姆医药品有限公司 代谢病症相关靶基因iRNA组合物和其使用方法
WO2025040002A1 (fr) * 2023-08-21 2025-02-27 苏州炫景生物科技有限公司 Oligonucléotide double brin ciblant un gène inhbe, conjugué, composition et utilisation associée
US20250075214A1 (en) * 2023-08-30 2025-03-06 Arrowhead Pharmaceuticals, Inc. RNAi Agents for Inhibiting Expression of Inhibin Subunit Beta E (INHBE), Pharmaceutical Compositions Thereof, and Methods of Use
CN120210203A (zh) * 2024-03-27 2025-06-27 施能康医药科技(苏州)有限公司 靶向抑制素亚单位βE的核酸及其用途

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116583291A (zh) * 2020-12-14 2023-08-11 雷杰纳荣制药公司 用抑制素亚基βE(INHBE)抑制剂治疗代谢病症和心血管疾病的方法
CN117716032A (zh) * 2021-07-21 2024-03-15 阿尔尼拉姆医药品有限公司 代谢病症相关靶基因iRNA组合物和其使用方法
WO2025040002A1 (fr) * 2023-08-21 2025-02-27 苏州炫景生物科技有限公司 Oligonucléotide double brin ciblant un gène inhbe, conjugué, composition et utilisation associée
US20250075214A1 (en) * 2023-08-30 2025-03-06 Arrowhead Pharmaceuticals, Inc. RNAi Agents for Inhibiting Expression of Inhibin Subunit Beta E (INHBE), Pharmaceutical Compositions Thereof, and Methods of Use
CN120210203A (zh) * 2024-03-27 2025-06-27 施能康医药科技(苏州)有限公司 靶向抑制素亚单位βE的核酸及其用途

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
SUGIYAMA MASAKAZU, KIKUCHI AKIHIRO, MISU HIROFUMI, IGAWA HIROBUMI, ASHIHARA MOTOOKI, KUSHIMA YOUICHI, HONDA KIYOFUMI, SUZUKI YOSHI: "Inhibin βE (INHBE) is a possible insulin resistance-associated hepatokine identified by comprehensive gene expression analysis in human liver biopsy samples", PLOS ONE, vol. 13, no. 3, 29 March 2018 (2018-03-29), pages e0194798, XP055898904, DOI: 10.1371/journal.pone.0194798 *

Also Published As

Publication number Publication date
CN120210203A (zh) 2025-06-27

Similar Documents

Publication Publication Date Title
CN113164509B (zh) 用于抑制17β-HSD 13型(HSD17B13)表达的RNAi试剂、其组合物和使用方法
TW202421169A (zh) 代謝疾患相關的標靶基因ｉＲＮＡ組成物及其使用方法
JP2026510791A (ja) インヒビンサブユニットベータｅ（ｉｎｈｂｅ）遺伝子の発現を阻害するための組成物および方法
US20200291406A1 (en) Preventive or therapeutic agent for fibrosis
TW202400193A (zh) 抑制跨膜絲胺酸蛋白酶６（ｔｍｐｒｓｓ６）表現的組成物及方法
EP4665852A1 (fr) Agents d'arn double brin modifiés
CN120210203A (zh) 靶向抑制素亚单位βE的核酸及其用途
WO2025193754A2 (fr) Compositions et méthodes d'inhibition de l'expression de gènes de la sous-unité bêta de l'inhibine (inhbe)
WO2025223537A1 (fr) Arndb, utilisation associée et procédé de préparation s'y rapportant
WO2025002247A1 (fr) Acide nucléique ciblant la protéine 3 de type angiopoïétine humaine et son utilisation
WO2026037221A1 (fr) Réactif d'arni ciblant le complément c3 et son utilisation
WO2025201156A1 (fr) Acide nucléique ciblant le facteur xi de coagulation et utilisation associée
WO2011074652A1 (fr) Acide nucléique capable d'inhiber l'expression de hif-2a
WO2024260380A1 (fr) Acide nucléique ciblant la lipoprotéine a et son utilisation
WO2024255749A1 (fr) Acide nucléique ciblant l'angiotensinogène et son utilisation
WO2024260379A1 (fr) Acide nucléique ciblant la cétohexokinase et son utilisation
WO2024255747A1 (fr) Acide nucléique ciblant le composant 5 du complément et son utilisation
WO2024255748A1 (fr) ACIDE NUCLÉIQUE CIBLANT L'HYDROXYSTÉROÏDE 17-β DÉSHYDROGÉNASE 13 ET SON UTILISATION
WO2026067590A1 (fr) Molécule d'arn double brin modifiée
WO2026026983A1 (fr) Oligonucléotide ciblant le gène inhbe et son utilisation
HK40116803A (zh) 靶向血管紧张素原的核酸及其用途
WO2026026712A1 (fr) Arnsi pour inhiber l'expression du gene masp2 et son modificateur et son utilisation
WO2026092544A1 (fr) Oligonucléotide ciblant le gène mtarc1 et utilisation de celui-ci
WO2025168134A1 (fr) Oligonucléotide ciblant le gène du composant 3 du complément (c3) et son utilisation
US20260009037A1 (en) RNAi Agents for Inhibiting Expression of Mitochondrial Amidoxime Reducing Component 1 (MARC1), Pharmaceutical Compositions Thereof, and Methods of Use

Legal Events

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

Ref document number: 25778570

Country of ref document: EP

Kind code of ref document: A1