WO2022211095A1 - Ｆｇｆｒ３のアンチセンスオリゴヌクレオチド - Google Patents

Ｆｇｆｒ３のアンチセンスオリゴヌクレオチド Download PDF

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WO2022211095A1
WO2022211095A1 PCT/JP2022/016874 JP2022016874W WO2022211095A1 WO 2022211095 A1 WO2022211095 A1 WO 2022211095A1 JP 2022016874 W JP2022016874 W JP 2022016874W WO 2022211095 A1 WO2022211095 A1 WO 2022211095A1
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nucleosides
compound
group
wing segment
nucleoside
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French (fr)
Japanese (ja)
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高尾 鈴木
正輝 山上
アジャヤラム セレスタ
幸介 千葉
忠士 梅本
駿 熊谷
夕貴 日高
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ルクサナバイオテク株式会社
日産化学株式会社
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Publication of WO2022211095A1 publication Critical patent/WO2022211095A1/ja

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    • 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
    • A61K31/7115—Nucleic acids or oligonucleotides having modified bases, i.e. other than adenine, guanine, cytosine, uracil or thymine
    • 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
    • A61K31/712—Nucleic acids or oligonucleotides having modified sugars, i.e. other than ribose or 2'-deoxyribose
    • 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
    • A61K31/7125—Nucleic acids or oligonucleotides having modified internucleoside linkage, i.e. other than 3'-5' phosphodiesters
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • 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

Definitions

  • the present invention relates to FGFR3 antisense oligonucleotides.
  • Achondroplasia is an autosomal overt inherited skeletal system disease with short stature of the limbs, and its cause is reported to be the Gly380Arg mutation in the FGFR3 gene located on chromosome 4p26.3.
  • ACH causes foramen magnum stenosis, ventricular enlargement/hydrocephalus due to poor perfusion of cerebrospinal fluid, and spinal canal stenosis. About 250,000 people worldwide are said to be affected by ACH, and the incidence is reported to be 1 in 10,000 to 30,000 live births (see, for example, Non-Patent Document 2).
  • FGFR3 signals suppress hypertrophy and proliferation of chondrocytes.
  • FGFR3 gene mutations that cause ACH are gain-of-function mutations. The differentiation of chondrocytes and the production and proliferation of cartilage matrix are inhibited, resulting in abnormal endochondral ossification, growth failure of long bones, hypoplasia of the base of the skull, and the like.
  • As therapeutic agents for ACH CNP (C-type natriuretic peptide) derivatives, soluble FGFR3 receptors, FGFR3 signal low-molecular-weight inhibitors, and the like are under development (see, for example, Non-Patent Document 3).
  • An object of the present invention is to provide a novel antisense oligonucleotide that inhibits the expression of the FGFR3 gene.
  • the present inventors have made extensive studies to discover compounds having an antisense effect on FGFR3, and have found that the compounds of the present invention have an excellent FGFR3 gene expression inhibitory action, leading to the completion of the present invention. That is, the present invention is characterized by the following.
  • An antisense oligonucleotide compound that inhibits the expression of FGFR3, wherein the antisense oligonucleotide compound comprises nucleobase positions 388-410, 671-688, 726-745, 860-887, 886- 915, 924-957, 1106-1130, 1157-1174, 1195-1210, 1216-1231, 1238-1256, 1276-1294, 1574-1588, 1622-1647, 1648-1662, 1730-1750, 1750-1771, 1779-1795, 1822-1836, 1850-1870, 1991-2005, 2017-2058, 2083-2112, 2189-2203, 2250-2264, 2528-2542, 2711-2726, 2782-2804, 2968-2987, 3203- 3217, 3558-3572, 3656-3670, 3712-3733, 3823-3837, 3920-3954, 4007-4023, 4040-4062, 4094-4116, 4164-4181, 4203-4217, or 42
  • an antisense oligonucleotide compound that inhibits expression of FGFR3, wherein the antisense oligonucleotide compound comprises nucleobase positions 388-410, 671-686, 726-743, 860-887, 886- 915, 929-956, 1106-1130, 1157-1174, 1196-1210, 1216-1231, 1238-1256, 1276-1292, 1574-1588, 1622-1647, 1648-1662, 1734-1749, 1750-1771, 1779-1795, 1822-1836, 1850-1866, 1991-2005, 2017-2058, 2083-2112, 2189-2203, 2250-2264, 2528-2542, 2711-2726, 2782-2804, 2968-2987, 3203- 3217, 3558-3572, 3656-3670, 3712-3733, 3823-3837, 3920-3954, 4007-4023, 4040-4062, 4094-4116, 4164-4181, 4203-4217, or 4245
  • the antisense oligonucleotide compound comprises nucleobase position numbers 388-402, 389-403, 390-404, 391-405, 392-406, 393-407, 394-408, 395-409, 396 of the nucleobases of SEQ ID NO:1.
  • Said antisense oligonucleotide compound comprises SEQ. 23, 27, 28, 29, 30, 31, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 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, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 92, 94, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 115, 116, 117, 118, 119, 120, 121, 122, 123, 126
  • Said antisense oligonucleotide compound comprises SEQ. 23, 27, 28, 29, 30, 31, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 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, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 92, 94, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 115, 116, 117, 118, 119, 120, 121, 122, 123, 126, 127, 128, 129, 130, 131, 132,
  • the antisense oligonucleotide compound comprises nucleobase position numbers 673-687, 674-688, 731-745, 924-938, 925-939, 926-940, 927-941, 928-942, 943 of the nucleobases of SEQ ID NO:1.
  • Said antisense oligonucleotide compounds have the 1. having any one nucleic acid base sequence selected from the group consisting of 267, 268, 269 and 271; or 6.
  • the antisense oligonucleotide compound comprises at least one 2'-4'-bridged nucleoside; to 7.
  • an antisense oligonucleotide compound that inhibits expression of FGFR3, wherein the antisense oligonucleotide compound comprises nucleobase positions 874-899, 948-962, 1213-1229, 1299-1314, 1748- 1763, 2693-2707, 4048-4071, or 4251-4270 consisting of 12 to 30 nucleosides complementary to at least part of the nucleobase sequence shown in and at least 90% complementary to at least part of the nucleobase sequence of SEQ ID NO: 1, at least 1 8. contains two 2'-4'-bridged nucleosides; A compound or a pharmacologically acceptable salt thereof according to. 11.
  • the antisense oligonucleotide compound comprises nucleobase position numbers 874-888, 875-889, 876-890, 877-891, 882-896, 883-897, 884-898, 885-899, 948 of the nucleobases of SEQ ID NO:1.
  • Said antisense oligonucleotide compound comprises SEQ. 153, 154, 202, 203, 204, 205, 206, 207, 215, 216, 217, 218, 222, 223, 224 and 225 10. having any one nucleic acid base sequence selected from the group consisting of or 11.
  • a compound or a pharmacologically acceptable salt thereof according to. 13 comprises nucleobase position numbers 675-689, 676-690, 944-958, 945-959, 946-960, 947-961, 949-963, 950-964, 951 of the nucleobases of SEQ ID NO:1.
  • ⁇ 965, 952-966, 953-967, 1205-1219, 1211-1225, or 1212-1226 9. consists of 12 to 30 nucleosides complementary to at least part of the nucleic acid base sequence shown in; A compound or a pharmacologically acceptable salt thereof according to. 14. Said antisense oligonucleotide compounds have the 9 or 13, having any one nucleic acid base sequence selected from the group consisting of A compound or a pharmacologically acceptable salt thereof according to.
  • the 2'-4'-bridged nucleoside is at least one selected from the group consisting of LNA, ENA, cEt, AmNA, scpBNA and GuNA; to 14.
  • the antisense oligonucleotide compound comprises a gap segment, a 5' wing segment and a 3' wing segment;
  • the gap segment consists of 5 to 20 nucleosides independently selected from the group consisting of deoxyribonucleosides and sugar-modified nucleosides and comprises at least two deoxyribonucleosides, and the 5' and 3' ends of the gap segment are , a deoxyribonucleoside or a 5'-modified nucleoside
  • the 5' wing segment consists of 1 to 10 nucleosides independently selected from the group consisting of deoxyribonucleosides and sugar-modified nucleosides, wherein the nucleoside at the 3' end of the 5' wing segment is a sugar-modified nucleoside; and linked to the 5' end of the gap segment;
  • the 3' wing segment consists of 1 to 10 nucleosides independently selected from the group consisting of deoxyribonucleo
  • the 5' end of said gap segment is a deoxyribonucleoside or a 5' modified nucleoside, and the 3' end of said gap segment is a deoxyribonucleoside.
  • the antisense oligonucleotide compound comprises a gap segment, a 5' wing segment and a 3' wing segment;
  • the gap segment consists of 5 to 20 nucleosides independently selected from the group consisting of deoxyribonucleosides and sugar-modified nucleosides and comprises at least two deoxyribonucleosides, and the 5' and 3' ends of the gap segment are , is a deoxyribonucleoside
  • the 5' wing segment consists of 1 to 10 nucleosides independently selected from the group consisting of deoxyribonucleosides and sugar-modified nucleosides, wherein the nucleoside at the 3' end of the 5' wing segment is a sugar-modified nucleoside; and linked to the 5' end of the gap segment;
  • the 3' wing segment consists of 1 to 10 nucleosides independently selected from the group consisting of deoxyribonucleosides and sugar-modified nucleoside
  • said gap segment consists of 8-15 nucleosides independently selected from the group consisting of deoxyribonucleosides and sugar modified nucleosides; said 5′ wing segment and said 3′ wing segment each consist of 2 to 5 nucleosides independently selected from the group consisting of deoxyribonucleosides and sugar modified nucleosides; 15. At least one of said 5' wing segment and said 3' wing segment comprises at least one 2'-4'-bridged nucleoside; to 18. A compound according to any one of or a pharmacologically acceptable salt thereof. 20.
  • said gap segment consists of 9-13 nucleosides independently selected from the group consisting of deoxyribonucleosides and sugar modified nucleosides; said 5′ wing segment and said 3′ wing segment each consist of 3 nucleosides independently selected from the group consisting of deoxyribonucleosides and sugar modified nucleosides; 18. At least one of said 5' wing segment and said 3' wing segment comprises at least one 2'-4'-bridged nucleoside; A compound or a pharmacologically acceptable salt thereof according to.
  • the sugar-modified nucleoside is at least one selected from the group consisting of 2'-4'-bridged nucleosides, 2'-modified nucleosides, and 5'-modified nucleosides. to 20. A compound according to any one of or a pharmacologically acceptable salt thereof. 22. 16. The sugar-modified nucleoside is at least one selected from the group consisting of 2'-4'-bridged nucleosides and 2'-modified nucleosides. to 20. A compound according to any one of or a pharmacologically acceptable salt thereof. 23.
  • the 2' modified nucleoside is at least one selected from the group consisting of 2'-O-Me nucleoside, 2'-O-MCE nucleoside, 2'-O-MOE nucleoside, and 2'-O-NMA nucleoside. Yes, 22. A compound or a pharmacologically acceptable salt thereof according to. 24. 21.
  • the 5'-modified nucleoside is a 5'-CP nucleoside; A compound or a pharmacologically acceptable salt thereof according to.
  • the 5' wing segment and the 3' wing segment each independently comprise at least one 2'-4'-bridged nucleoside; to 24. A compound according to any one of or a pharmacologically acceptable salt thereof.
  • the 5' wing segment and the 3' wing segment each independently comprise at least one AmNA; to 25. A compound according to any one of or a pharmacologically acceptable salt thereof.
  • 27. the nucleosides of the 5' wing segment are independently selected from 2'-4'-bridged nucleosides; 16.
  • the nucleosides of said 3' wing segment are independently selected from the group consisting of deoxyribonucleosides and 2'-4'-bridged nucleosides; to 26.
  • nucleosides of the 5' wing segment are independently selected from the group consisting of LNA, AmNA and GuNA; 16.
  • nucleosides of said 3' wing segment are independently selected from the group consisting of deoxyribonucleosides, LNA, AmNA, scpBNA and GuNA; to 27.
  • the nucleosides of the 5' wing segment are independently selected from the group consisting of AmNA and GuNA; 16.
  • the nucleosides of said 3' wing segment are independently selected from the group consisting of deoxyribonucleosides, LNA, AmNA, scpBNA and GuNA; to 27. A compound according to any one of or a pharmacologically acceptable salt thereof.
  • the nucleosides of said gap segment are independently selected from the group consisting of deoxyribonucleosides and 5'-CP nucleosides; to 29. A compound according to any one of or a pharmacologically acceptable salt thereof.
  • the internucleoside linkages contained in said gap segment, 5' wing segment and 3' wing segment are each independently selected from phosphorothioate linkages and phosphodiester linkages and comprise at least one phosphorothioate linkage; to 30.
  • the internucleoside linkages contained in said gap segment are phosphorothioate linkages.
  • a compound or a pharmacologically acceptable salt thereof according to. 33. 16 each cytosine of the sugar-modified nucleosides contained in the 5' and 3' wing segments is replaced with 5-methylcytosine; to 32.
  • the pharmacologically acceptable salt is a sodium salt; ⁇ 33. The salt according to any one of 35. 1. ⁇ 33.
  • a medicament comprising as an active ingredient the compound or pharmacologically acceptable salt according to any one of 36. 1. ⁇ 33.
  • An expression inhibitor of the FGFR3 gene comprising the compound or pharmacologically acceptable salt according to any one of the above as an active ingredient. 38. 1. ⁇ 33.
  • a drug for the treatment, prevention and/or improvement of achondroplasia comprising as an active ingredient a compound or a pharmacologically acceptable salt according to any one of
  • novel antisense oligonucleotides that have excellent gene expression inhibitory activity and are particularly useful for the treatment, improvement and/or prevention of achondroplasia.
  • 1 is a graph showing the effect of antisense oligonucleotides according to this embodiment on the expression levels of human FGFR1 and FGFR3 in HepG2 cells.
  • 4 is a graph showing the effect on the expression level of mouse FGFR3 in cartilage tissue of FVB/NJcl mice to which antisense oligonucleotides according to this embodiment were administered.
  • 4 is a graph showing the effect on the expression level of mouse FGFR3 in cartilage tissue of FVB/NJcl mice to which antisense oligonucleotides according to this embodiment were administered.
  • 4 is a graph showing the effect on the expression level of mouse FGFR3 in cartilage tissue of FVB/NJcl mice to which antisense oligonucleotides according to this embodiment were administered.
  • 4 is a graph showing the effect on the expression level of mouse FGFR3 in cartilage tissue of FVB/NJcl mice to which antisense oligonucleotides according to this embodiment were administered.
  • antisense oligonucleotides or antisense oligonucleotide compounds are used interchangeably and are sometimes referred to as "ASO.”
  • Nucleic acids in nature are most basically composed of adenosine (A), thymidine (T) (or uridine (U)), cytidine (C) and guanosine (G).
  • Their basic nucleic acids are often referred to as AT(U)GC and the like. Therefore, in this specification, when a sequence is indicated by "nucleobase sequence” or “sequence number”, it is basically a sequence of A, T, G, C, and U with respect to, for example, the sequence of the FGFR3 gene.
  • the nucleic acids constituting the ASO of the present invention include basic nucleic acids (AT(U)CG) as well as structurally modified versions thereof.
  • Optionally substituted means unsubstituted or substituted.
  • nucleoside is a term well known to those skilled in the art, and is generally understood to be a molecule in which a sugar and a nucleobase are bound, and which can be a unit constituting a nucleic acid. As used herein, nucleoside is a concept in a broader sense and includes ribonucleosides, deoxyribonucleosides and sugar-modified nucleosides described below. The nucleobases may be modified.
  • “Ribonucleoside” means a molecule having a nucleobase at the carbon atom at position 1 of ribose.
  • a ribonucleoside in the present invention may be a naturally occurring ribonucleoside or a ribonucleoside in which the nucleobase portion of a naturally occurring ribonucleoside is modified.
  • a single ribonucleoside may be modified in combination of multiple types.
  • the modified ribonucleosides are, for example, Journal of Medicinal Chemistry, 2016, 59, pp 9645-9667, Medicinal Chemistry Communication, 2014, 5, pp 1454-1471, Future Medicinal Chemistry, 2011, 3, pp 339-365, It is described in International Publication No. 2018/155450 and the like.
  • Deoxyribonucleoside means a molecule in which the 2' hydroxy group of ribonucleoside is replaced with a hydrogen atom.
  • Deoxyribonucleosides in the present invention may be naturally occurring deoxyribonucleosides or deoxyribonucleosides obtained by modifying the nucleobase portion of naturally occurring deoxyribonucleosides.
  • a single deoxyribonucleoside may be modified in combination of multiple types.
  • the modified deoxyribonucleosides are, for example, Journal of Medicinal Chemistry, 2016, 59, pp 9645-9667, Medicinal Chemistry Communication, 2014, 5, pp 1454-1471, Future Medicinal Chemistry, 2011, 3, pp 339-365, It is described in International Publication No. 2018/155450 and the like.
  • a "modified sugar” is (Z1) molecules in which the ribose or 2-deoxyribose is partially substituted by one or more substituents, (Z2) pentasaccharides or hexoses different from ribose and 2-deoxyribose (e.g., hexitol, threose, etc.), (Z3) Whole ribose or 2-deoxyribose, or a tetrahydrofuran ring thereof, binds a 5- to 7-membered saturated or unsaturated ring (e.g., cyclohexane, cyclohexene, morpholine, etc.), or a 5- to 7-membered ring via hydrogen bonding molecules that have been replaced by substructures that can form (e.g., peptide structures), or (Z4) refers to a molecule in which ribose or 2-deoxyribose is replaced with alkylene glycol having 2 to 6 carbon
  • Modified sugars include “2-modified sugars”, “2-4-linked sugars” and “5-modified sugars” described below.
  • modified sugars and sugar-modified nucleosides described later include, for example, JP-A-10-304889, WO-2005/021570, JP-A-10-195098, JP-A-2002-521310, and WO-2007. / 143315, International Publication No. 2008/043753, International Publication No. 2008/029619, International Publication No. 2008/049085 and International Publication No.
  • modified sugars that are partially substituted with one substituent include one or more (preferably one or two) substitutions of (i) or (ii) below at any position of the sugar moiety.
  • Ribose or 2-deoxyribose substituted with groups may be mentioned.
  • halogen atoms C 1-6 alkoxy groups, haloC 1-6 alkoxy groups, mono- or di-C 1-6 alkylamino groups, 5-10 membered heterocyclic groups, carboxy groups, carbamoyl groups, and N - a C 1-6 alkyl group substituted with at least one selected from the group consisting of substituted carbamoyl groups;
  • the N-substituted carbamoyl group includes an N-methyl-carbamoyl group and an N-ethyl-carbamoyl group, where the methyl group and the ethyl group of the N-methyl-carbamoyl group and N-ethyl-carbamoyl group
  • the group may be optionally substituted with a 5-10 membered heterocyclic group or a mono- or di-C 1-6 alkylamino group.
  • N-substituted carbamoyl groups include N-methylcarbamoyl, N-ethylcarbamoyl, N-dimethylaminoethyl-carbamoyl, N-(morpholinoethyl)carbamoyl, and N-(2-pyridylethyl)carbamoyl. group, N-((benzimidazol-1-yl)ethyl)carbamoyl group, and the like.
  • “Sugar-modified nucleoside” means a molecule having the aforementioned "modified sugar” in place of the sugar moiety of a deoxyribonucleoside or ribonucleoside. For example, it includes “2'-modified nucleosides” and “2'-4' bridged nucleosides” described below, and also includes “5'-modified nucleosides.”
  • sugar-modified nucleosides also include molecules in which the modified sugar and a nucleobase are linked via a methylene chain or the like.
  • 2-modified sugar means a non-bridged sugar in which the oxygen atom or carbon atom at the 2-position of ribose is modified, such as "2-O-Me”, “2-O-MOE”, “2-O -MCE”, “2-O-NMA”, “2-O-AP”, “2-F”, “2-DMAECE”, “2-MorECE”, “2-PyECE” and “2-BimECE” contain. Positions 4 and 5 of the "2-modified sugar" are preferably unmodified.
  • “2'-modified nucleoside” means a molecule having a nucleobase at position 1 of the "2-modified sugar", for example, "2'-O-Me nucleoside”, “2'-O-MOE nucleoside”, “2'-O-MCEnucleoside”,”2'-O-NMAnucleoside”,”2'-O-APnucleoside”,”2'-Fnucleoside”,”2'-DMAECEnucleoside”,”2'-MorECE nucleosides”, “2′-PyECE nucleosides” and “2′-BimECE nucleosides”.
  • 2-O-Me also called 2-O-methyl
  • 2-O-Me nucleoside also called 2'-O-methyl nucleoside
  • 2-O-MOE also called 2-O-methoxyethyl
  • 2-O-methoxyethyl means a sugar in which the hydroxy group at position 2 of ribose has been replaced with a 2-methoxyethyloxy group.
  • 2-O-MOE nucleoside also referred to as 2'-O-methoxyethyl nucleoside
  • 2-O-MCE also called 2-O-methylcarbamoylethyl
  • 2-O-methylcarbamoylethyl means a sugar in which the hydroxy group at position 2 of ribose has been replaced with a methylcarbamoylethyloxy group.
  • 2'-O-MCE nucleoside also referred to as 2'-O-methylcarbamoylethyl nucleoside
  • 2-O-NMA means a sugar in which the hydroxy group at position 2 of ribose has been replaced with a [2-(methylamino)-2-oxoethyl]oxy group.
  • 2'-O-NMA nucleoside means a molecule having the nucleobase at position 1 of "2-O-NMA”.
  • 2-O-AP means a sugar in which the hydroxy group at position 2 of ribose has been replaced with a 3-aminopropyloxy group.
  • 2'-O-AP nucleoside means a molecule having the nucleobase at position 1 of "2-O-AP”.
  • 2-F means a sugar in which the hydroxy group at position 2 of ribose is replaced with a fluorine atom.
  • 2'-F nucleoside means a molecule with a nucleobase at position 1 of "2-F”.
  • 2-DMAECE 2-MorECE
  • 2-PyECE 2-PyECE
  • 2-BimECE are structures in which the hydroxy group at the 2-position of ribose is represented by the following DMAECE, MorECE, PyECE, and BimECE, respectively. is a sugar replaced by In the structure below, the wavy line indicates the bonding position with the carbon atom to which the 2-position hydroxy group of ribose is bonded.
  • a "2-4 bridged sugar” means a sugar in which the bridging unit is substituted by two substitutions at positions 2 and 4 on ribose.
  • the bridging unit include a C 2-6 alkylene group (the alkylene group is unsubstituted or substituted with one or more substituents selected from the group consisting of a halogen atom, an oxo group and a thioxo group.
  • alkylene group and one or two methylene groups of said alkylene group are either unsubstituted or independently —O—, —NR 1 — (R 1 is a hydrogen atom, a C 1-6 alkyl group or haloC 1 -6 alkyl group) and replaced with a group selected from the group consisting of -S-).
  • R 1 is a hydrogen atom, a C 1-6 alkyl group or haloC 1 -6 alkyl group
  • the 5-position of the "2-4 bridged sugar" is preferably unmodified.
  • a "2'-4' bridged nucleoside"(2',4'-BNA) means a molecule having a nucleobase at position 1 of said "2-4 bridged sugar".
  • ⁇ -D-methyleneoxy (4′-CH 2 -O-2′) BNA also called LNA (Locked Nucleic Acid®)
  • ⁇ -L-methyleneoxy (4′-CH 2 -O-2')BNA ethyleneoxy (4'-(CH 2 ) 2 -O-2')BNA, also called ENA
  • (4 ⁇ -D-thio'-CH 2 -S-2')BNA oxyamino (4′-CH 2 —ON(R 11 )-2′) BNA
  • R 11 is a hydrogen atom or methyl
  • 2′,4′-BNA NC 4′-CH 2 —N(R 12 )-O-2′)BNA R 12 is a hydrogen atom or methyl
  • 2′,4′-BNA COC 3′-amino-2
  • Base is a nucleobase.
  • AmNA[Me] is AmNA wherein said R13 is methyl.
  • GuNA[tBu] is GuNA wherein R 15 is t-butyl and R 16 is H.
  • "5-modified sugar” means a non-crosslinked sugar modified at the 5-position of 2-deoxyribose, and includes “5-CP", “5-methyl” and “5-aminopropyl”. Positions 2 and 4 of the "5-modified sugar” are preferably unmodified.
  • "5'-modified nucleoside” means a molecule having a nucleobase at position 1 of the "5-modified sugar”, such as "5'-CP nucleoside”, “5'-methyl nucleoside” and "5'-aminopropyl nucleoside”.
  • “5-CP” is a sugar in which the 5-position of 2-deoxyribose is substituted with two methyl groups, which together form cyclopropane.
  • a “5'-CP nucleoside” is a molecule represented by the following structural formula. In the formula, Base is a nucleobase.
  • “5-Methyl” is a sugar in which the 5-position of 2-deoxyribose is substituted with a methyl group.
  • “5-Aminopropyl” is a sugar in which the 5-position of 2-deoxyribose is substituted with a 3-aminopropyl group.
  • “5'-methyl nucleoside” and “5'-aminopropyl nucleoside” refer to molecules having the nucleobase at position 1 of "5-methyl” and "5-aminopropyl” respectively.
  • the bond between the carbon atom at the 1' position and the nucleobase is an ⁇ -glycosidic bond and a ⁇ - glycosidic bond, usually a ⁇ -glycosidic bond. Therefore, ⁇ -D-methyleneoxy BNA is usually used as LNA.
  • n- means normal, "s-” means secondary, and "t-" means tertiary.
  • Halogen atom means a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.
  • C 1-6 alkyl group means a linear or branched saturated hydrocarbon group having 1 to 6 carbon atoms, such as methyl group, ethyl group, n-propyl group, isopropyl group, n -butyl group, isobutyl group, s-butyl group, t-butyl group, n-pentyl group, isopentyl group, neopentyl group, n-hexyl group and isohexyl group.
  • halo C 1-6 alkyl group means a group in which a hydrogen atom at any position in the above “C 1-6 alkyl group” is substituted with one or more above-mentioned “halogen atoms”.
  • C 1-6 alkylene group means a divalent group obtained by removing one hydrogen atom at any position from a linear or branched saturated hydrocarbon group having 1 to 6 carbon atoms, for example Examples include methylene group, ethylene (ethanediyl) group, propane-1,3-diyl (trimethylene) group, propane-2,2-diyl group, 2,2-dimethyl-propane-1,3-diyl group, hexane- 1,6-diyl (hexamethylene) group, 3-methylbutane-1,2-diyl group and the like.
  • C 2-6 alkylene group means a linear or branched divalent group having 2 to 6 carbon atoms among the above “C 1-6 alkylene groups”, examples of which include methylene It is the same as the above “C 1-6 alkylene group” except for the group.
  • C 1-6 alkoxy group means a group in which the above “C 1-6 alkyl group” is bonded to an oxy group.
  • halo C 1-6 alkoxy group means a group in which a hydrogen atom at an arbitrary position in the above “C 1-6 alkoxy group” is substituted with one or more above-mentioned “halogen atoms”.
  • “Mono- or di-C 1-6 alkylamino group” means a group in which one hydrogen atom of an amino group is replaced with one "C 1-6 alkyl group", or two hydrogen atoms of an amino group are means a group replaced by two identical or different "C 1-6 alkyl groups", for example, methylamino, ethylamino, propylamino, isopropylamino, butylamino, dimethylamino, diethylamino, dipropylamino, dibutylamino , and N-ethyl-N-methylamino groups.
  • 3- to 6-membered ring means a monocyclic saturated or unsaturated hydrocarbon ring having 3 to 6 carbon atoms, such as cyclopropane, cyclobutane, and cyclohexane.
  • 5- to 10-membered heterocyclic group is a 5- to 10-membered monocyclic or condensed ring system containing 1 to 4 heteroatoms selected from nitrogen atoms, sulfur atoms and oxygen atoms in addition to carbon atoms as ring-constituting atoms It means a polycyclic aromatic or non-aromatic heterocyclic group.
  • Preferable examples of the "5- to 10-membered heterocyclic group" include thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazolyl, tetrazolyl, triazinyl, benzo Thiophenyl, benzofuranyl, benzoimidazolyl, benzoxazolyl, benzoisoxazolyl, benzothiazolyl, benzoisothiazolyl, benzotriazolyl, imidazopyridinyl, thienopyridinyl, pyrrolopyridinyl, pyrazolopyridinyl, oxazolopyridinyl , thiazolopyridinyl, imidazopyrazinyl,
  • nucleobase is a purine base or a pyrimidine base, and may be a naturally occurring nucleobase or a modified naturally occurring nucleobase.
  • Naturally occurring nucleobases herein include adenine (A), guanine (G), thymine (T), cytosine (C), uracil (U) and 5-methylcytosine (5).
  • the “nucleobase” includes naturally occurring nucleobases and "modified nucleobases" described later.
  • nucleobase modifications in the "modified nucleobase” herein include halogenation, methylation, ethylation, n-propylation, isopropylation, cyclopropylation, n-butylation, isobutylation, s- Butylation, t-butylation, cyclobutylation, hydroxylation, amination, thiolation, demethylation and the like.
  • the nucleobase in the nucleoside is more preferably at least one selected from the group consisting of adenine, guanine, thymine, cytosine, uracil, 5-methylcytosine, 5-hydroxycytosine and 8-aminoguanine, particularly preferably , adenine, guanine, thymine, cytosine, uracil and 5-methylcytosine.
  • 5-methylcytosine means cytosine with a methyl group at the 5-position.
  • 5-Hydroxycytosine means a cytosine with a hydroxy group at the 5-position.
  • 8-Aminoguanine means guanine with an amino group at the 8-position.
  • Nucleobase sequence means the sequence from the 5' side to the 3' side of the nucleobase of each nucleoside contained in the oligonucleotide.
  • Contiguous nucleobase means a sequence from the 5' side to the 3' side of a part of consecutive nucleobases in the above-mentioned “nucleobase sequence”.
  • Internucleoside linkage means a group or bond that forms a covalent bond between adjacent nucleosides in an oligonucleotide.
  • Internucleoside linkages include phosphodiester linkages and "modified internucleoside linkages” described below.
  • Modified internucleoside linkage means a modified phosphodiester linkage, e.g., phosphorothioate linkage, methylphosphonate linkage (including chiral-methylphosphonate linkage), methylthiophosphonate linkage, phosphorodithioate linkage, phosphoramidate bond, phosphorodiamidate bond, phosphoramidothioate bond, boranophosphate bond and the like. Also, Journal of Medicinal Chemistry, 2016, 59, pp 9645-9667, Medicinal Chemistry Communication, 2014, 5, pp 1454-1471, Future Medicinal Chemistry, 2011, 3, pp 339-365, etc. Examples are disclosed and can be used for modified phosphodiester linkages.
  • Oligonucleotide is a molecule having a structure in which two or more identical or different “nucleosides” are linked by the independently selected “internucleoside bond” (e.g., phosphodiester bond or modified phosphodiester bond).
  • internucleoside bond e.g., phosphodiester bond or modified phosphodiester bond.
  • DNA means a polynucleotide or oligonucleotide in which two or more of the same or different “deoxyribonucleosides” are linked by the independently selected “internucleoside linkages”.
  • RNA means a polynucleotide or oligonucleotide in which two or more of the same or different “ribonucleosides” are linked by the independently selected “internucleoside linkages”.
  • antisense effect means that the function of the target RNA is controlled by hybridizing the target RNA selected corresponding to the target gene and, for example, an oligonucleotide having a sequence complementary to its partial sequence.
  • hybridization inhibits the translation of the target RNA, a splicing function conversion effect such as exon skipping, and the target RNA is degraded by recognition of the hybridized portion. etc.
  • the target RNA is "FGFR3 mRNA".
  • ASO antisense oligonucleotide
  • ASO is an oligonucleotide that produces the antisense effect. Examples include, but are not limited to, DNA, gapmers, mixmers, and the like, and may be RNA or oligonucleotides designed to normally produce an antisense effect. ASOs of the invention are single-stranded antisense oligonucleotides.
  • Hybridize refers to the act of forming a duplex between oligonucleotides containing complementary sequences or portions thereof, and oligonucleotides containing complementary sequences or portions thereof forming duplexes means phenomenon.
  • “Complementary” means that two nucleobases can form Watson-Crick base pairs (natural base pairs) or non-Watson-Crick base pairs (Hoogsteen base pairs, etc.) through hydrogen bonding.
  • Two oligonucleotides or portions thereof can "hybridize” if their sequences are complementary. Although it is not necessary for two oligonucleotides or portions thereof to hybridize they must be perfectly complementary, the complementarity for two oligonucleotides or portions thereof to hybridize is preferably 70%. or more, more preferably 80% or more, and still more preferably 90% or more (for example, 95%, 96%, 97%, 98%, or 99% or more). Sequence complementarity is determined by utilizing computer programs that automatically identify subsequences of oligonucleotides. For example, GENETYX (manufactured by Genetics Inc.) is one such software.
  • Gapmer means an oligonucleotide containing a "gap segment", a "5' wing segment” and a “3' wing segment” described below.
  • a “gap segment” is a region containing "at least four consecutive nucleosides recognized by RNase H", and is not particularly limited as long as it contains four or more consecutive nucleosides and is recognized by RNase H, but said consecutive Nucleosides are preferably independently selected from deoxyribonucleosides and sugar-modified nucleosides.
  • the nucleosides at the 5' and 3' ends of the gap segment in the present invention are preferably 5' modified nucleosides or deoxyribonucleosides, more preferably deoxyribonucleosides or 5'-CP nucleosides.
  • 5' wing segment is a region linked to the 5' side of the gap segment and containing "at least one nucleoside” without the "at least four consecutive nucleosides recognized by RNase H";
  • the 3'-terminal nucleoside sugar moiety of the 5' wing segment is different from the 5'-terminal nucleoside sugar moiety of the gap segment.
  • Differences in sugar moieties confirm the boundaries of the 5' wing and gap segments.
  • the difference in sugar moieties is preferably determined by the presence or absence of modification at the 2-position of the corresponding sugar.
  • the "2-modified sugar” is a non-crosslinked sugar in which the oxygen atom or carbon atom at the 2-position of ribose is modified
  • the "2-4 bridged sugar” is defined as 2 Both are modified at the 2-position because they are sugars in which the bridging unit is substituted by two substitutions at the position and the 4-position.
  • the nucleosides at the 5' end of the gap segment are deoxyribonucleosides and the nucleosides at the 3' end of the 5' wing segment are sugar-modified nucleosides.
  • the sugar-modified nucleosides are 2'-4'-bridged nucleosides.
  • nucleoside at the 5′ end of the gap segment is a 5′-CP nucleoside and the nucleoside at the 3′ end of the 5′ wing segment is a 2′-4′-bridged nucleoside or a 2'-modified nucleoside.
  • the 3'-terminal nucleoside of the 5' wing segment is generally a sugar-modified nucleoside, preferably a 2'-4'-bridged nucleoside or a 2'-modified nucleoside, and more A 2'-4'-bridged nucleoside is preferred.
  • the 5' wing segment is not particularly limited as long as it satisfies the above definition, but the at least one nucleoside is preferably independently selected from deoxyribonucleosides and sugar-modified nucleosides, and includes at least one sugar-modified nucleoside, preferably contains at least one 2′-4′-bridged nucleoside or 2′-modified nucleoside, particularly preferably a 2′-4′-bridged nucleoside.
  • the "3' wing segment” is a region linked to the 3' side of the gap segment and containing "at least one nucleoside” without the "at least four consecutive nucleosides recognized by RNase H",
  • the 5'-terminal nucleoside sugar moiety of the 3' wing segment is different from the 3'-terminal nucleoside sugar moiety of the gap segment.
  • Differences in sugar moieties identify the boundary between the 3' wing segment and the gap segment.
  • the difference in sugar moieties is preferably determined by the presence or absence of modification at the 2-position of the corresponding sugar.
  • nucleosides at the 3' end of the gap segment are deoxyribonucleosides and the nucleosides at the 5' end of the 3' wing segment are sugar-modified nucleosides.
  • the sugar-modified nucleosides are 2'-4'-bridged nucleosides.
  • the nucleoside at the 3′ end of the gap segment is a 5′-CP nucleoside and the nucleoside at the 5′ end of the 3′ wing segment is a 2′-4′-bridged nucleoside or a 2'-modified nucleoside.
  • the 5'-terminal nucleoside of the 3' wing segment is generally a sugar-modified nucleoside, preferably a 2'-4'-bridged nucleoside or a 2'-modified nucleoside, and more A 2'-4'-bridged nucleoside is preferred.
  • the 3' wing segment is not particularly limited as long as it satisfies the above definition, but the at least one nucleoside is preferably independently selected from deoxyribonucleosides and sugar-modified nucleosides, and includes at least one sugar-modified nucleoside, preferably contains at least one 2′-4′-bridged nucleoside or 2′-modified nucleoside, particularly preferably a 2′-4′-bridged nucleoside.
  • the portion where the 2'-modified nucleoside or the 2'-4'-bridged nucleoside continues from the 5'-end is the 5'-wing segment, and the nucleoside at the 3'-end of the 5'-wing segment is 2'-modified.
  • the boundary connecting nucleosides or other nucleosides (deoxyribonucleosides, 5' modified nucleosides, ribonucleosides, etc.) other than 2'-4'-bridged nucleosides is the boundary between the 5' wing segment and the gap segment.
  • the portion where the 2'-modified nucleoside or the 2'-4'-bridged nucleoside continues from the 3'-end is the 3'-wing segment, and the nucleoside at the 5'-end of the 3'-wing segment is 2'-modified.
  • the boundary connecting nucleosides or other nucleosides (deoxyribonucleosides, 5'-modified nucleosides, ribonucleosides, etc.) other than 2'-4'-bridged nucleosides is the boundary between the 3' wing segment and the gap segment.
  • the 3'-terminal deoxyribonucleoside and the portion where the deoxyribonucleoside is followed by a 2'-modified nucleoside or a 2'-4'-bridged nucleoside are the 3'-wing segment, and the 5'-terminal of the 3'-wing segment.
  • nucleosides are connected to other nucleosides (deoxyribonucleosides, 5'-modified nucleosides, ribonucleosides, etc.) other than 2'-modified nucleosides or 2'-4'-bridged nucleosides.
  • the boundary between the 3' wing segment and the gap segment is be.
  • RNase H is generally known as a ribonuclease that recognizes a double strand in which DNA and RNA hybridize in vivo and cleaves the RNA to generate single-stranded DNA.
  • RNase H recognizes not only double strands in which DNA and RNA hybridize, but also double strands in which at least one of the nucleobase moiety, phosphodiester linkage moiety and sugar moiety of at least one of DNA and RNA is modified. obtain. For example, it can also recognize duplexes in which DNA modified with phosphorothioate linkages and RNA are hybridized.
  • DNA can be recognized by RNaseH when hybridized with RNA.
  • RNA can also be cleaved by RNase H when hybridized with DNA.
  • typical examples include oligonucleotides in which the phosphodiester binding portion of DNA is modified with phosphorothioate.
  • modifications of DNA and/or RNA that can be recognized by RNaseH are, for example, Nucleic Acids Research, 2014, 42, pp 5378-5389, Bioorganic & Medicinal Chemistry Letters, 2008, 18, pp 2296-2300, Molecular BioSystems, 2009, 5, pp 838-843, Nucleic Acid Therapeutics, 2015, 25, pp 266-274, The Journal of Biological Chemistry, 2004, 279, pp 36317-36326.
  • the RNase H used in the present invention is preferably mammalian RNase H, more preferably human RNase H, and particularly preferably human RNase H1.
  • At least 4 consecutive nucleosides recognized by RNaseH includes 4 or more consecutive nucleosides, and is not particularly limited as long as it is recognized by RNaseH. For example, “at least 4 consecutive deoxyribonucleosides", etc. are mentioned.
  • the number of nucleosides constituting "at least four consecutive nucleosides recognized by RNaseH” is, for example, 5 to 20, preferably 5 to 15, more preferably 8 to 14, and even more preferably. is 9 to 13, particularly preferably 9. Whether a given at least four consecutive nucleosides are "at least four consecutive nucleosides recognized by RNase H" can be determined by those skilled in the art from the structure of the sugar portion of the consecutive nucleosides.
  • FGFR3 (fibroblast growth factor receptor 3) means any protein of FGFR3.
  • FGFR3 mRNA means mRNA encoding the FGFR3 protein.
  • FGFR3 nucleic acid means any nucleic acid that encodes FGFR3, including, for example, “FGFR3 mRNA”.
  • FGFR3 mRNA is represented by SEQ ID NO: 1, for example.
  • FGFR3 mRNA has nucleosides linked together by phosphodiester bonds, and thymine is usually replaced with uracil.
  • FGFR3 mRNA does not have any other modified sugars, modified nucleobases, or modified internucleoside linkages.
  • Gene expression refers to the conversion of gene code information into structures or functions in cells. Such structures include, but are not limited to, transcription and translation products (mRNA, pre-mRNA, proteins, etc.).
  • the ASO of the present invention targets FGFR3 mRNA.
  • the ASO of the present invention does not need to hybridize with the entire FGFR3 mRNA, and usually hybridizes with at least a portion of it. For example, by hybridizing a part of FGFR3 mRNA with an oligonucleotide having a sequence complementary to a partial sequence of FGFR3 mRNA (such as a gapmer or an oligonucleotide usually designed to produce an antisense effect) , which regulates the expression of the FGFR3 gene.
  • an oligonucleotide having a sequence complementary to a partial sequence of FGFR3 mRNA such as a gapmer or an oligonucleotide usually designed to produce an antisense effect
  • the complementarity between the ASO nucleobase sequence of the present invention and the partial sequence of FGFR3 mRNA is 90% or more (eg, 95%, 96%, 97%, 98% or 99% or more).
  • the sequences do not need to be completely complementary, but it is even more preferable that they are completely complementary. .
  • ASOs of the invention have a gap segment, a 5' wing segment and a 3' wing segment.
  • the gap segment consists of at least 5 nucleosides independently selected from the group consisting of deoxyribonucleosides, ribonucleosides and sugar-modified nucleosides and includes at least 2 deoxyribonucleosides, the 3' and 5' ends of which are Each independently is a deoxyribonucleoside.
  • a gap segment preferably comprises "at least 4 consecutive deoxyribonucleosides".
  • the number of nucleosides contained in the gap segment is preferably 5 to 20, more preferably 5 to 15, still more preferably 8 to 15, even more preferably 9 to 13, particularly preferably is 9.
  • the nucleosides contained in the gap segment are preferably independently selected from the group consisting of deoxyribonucleosides and sugar-modified nucleosides, more preferably deoxyribonucleosides.
  • Sugar-modified nucleosides contained in the gap segment may be any of 2'-modified nucleosides, 2'-4'-bridged nucleosides and 5'-modified nucleosides, but are preferably 2'-modified nucleosides, more preferably 2'- selected from O-Me nucleosides, 2'-O-MOE nucleosides, 2'-O-AP nucleosides, 2'-F nucleosides, 2'-O-NMA nucleosides and 2'-O-MCE nucleosides, more preferably 2 '-O-Me nucleoside.
  • the sugar-modified nucleoside contained in the gap segment is preferably a 5'-modified nucleoside, more preferably a 5'-CP nucleoside.
  • the positions of the 2' modified nucleosides and 2'-4' bridging nucleosides in the gap segment are not otherwise limited except at the 3' and 5' ends of the gap segment.
  • the position of the 5'-modified nucleoside in the gap segment is not particularly limited, but the 5'-modified nucleoside is preferably included on the 5' side of the gap segment.
  • the number of sugar-modified nucleosides contained in the gap segment is preferably 1-3.
  • the number of 2'-O-Me nucleosides contained in the gap segment is particularly preferably one.
  • the gap segment contains one or more 5'-CP nucleosides the number of 5'-CP nucleosides contained in the gap segment is particularly preferably 1-3.
  • the arrangement of the sugar modification in the gap segment is preferably selected from the group consisting of DDDDDDDD, ZDDDDDDDD, ZZDDDDDDDD, ZZZDDDDDD, DZDDDDDDDD, DDZDDDDDD, DDDZDDDDDD and DDDDZDDDDD, particularly preferably selected from the group consisting of DDDDDDDD, ZDDDDDDZDDDZ be done.
  • Z is a 5'-CP nucleoside
  • D is a deoxyribonucleoside
  • the left side indicates the 5' side and the right side indicates the 3' side.
  • the internucleoside linkages contained in the gap segment are preferably independently selected from phosphodiester linkages and modified phosphodiester linkages. and more preferably at least one independently selected from a phosphodiester bond and a phosphorothioate bond.
  • the internucleoside linkages contained in the gap segment preferably contain at least one phosphorothioate linkage, more preferably 50% or more are phosphorothioate linkages, more preferably 75% or more are phosphorothioate linkages, and 80% More preferably, more than 90% are phosphorothioate bonds, even more preferably 90% or more are phosphorothioate bonds, and most preferably all are phosphorothioate bonds.
  • the 5' internucleoside bond of the 5' modified nucleoside is preferably a phosphodiester bond or a phosphorothioate bond, more preferably a phosphodiester bond.
  • the internucleoside linkage is preferably a phosphodiester linkage.
  • the internucleoside linkage is preferably a phosphodiester linkage or a phosphorothioate linkage, more preferably a phosphorothioate linkage.
  • the 5' wing segment consists of at least one nucleoside independently selected from the group consisting of deoxyribonucleosides, ribonucleosides and sugar-modified nucleosides, provided that its 3' end that joins the gap segment is a sugar-modified nucleoside. Yes and does not include "at least 4 consecutive nucleosides recognized by RNase H".
  • the 3' wing segment consists of at least one nucleoside independently selected from the group consisting of deoxyribonucleosides, ribonucleosides and sugar-modified nucleosides, provided that its 5' end that joins the gap segment is a sugar-modified nucleoside.
  • At least one of the 5' and 3' wing segments preferably includes at least one 2'-4'-bridged nucleoside.
  • the 5' and 3' wing segments preferably do not contain "at least 4 consecutive deoxyribonucleosides".
  • each wing segment is preferably 1 to 10, more preferably 1 to 7, even more preferably 2 to 6, still more preferably 2 to 5. , still more preferably 2 to 4, and particularly preferably 3.
  • Each wing segment preferably contains a nucleoside with increased affinity for the partial sequence of FGFR3 mRNA or a nucleoside with increased resistance to nucleolytic enzymes due to substitution or the like.
  • the nucleosides contained in each wing segment are more preferably independently selected from sugar-modified nucleosides and deoxyribonucleosides, more preferably independently selected from 2'-4' bridged nucleosides, 2'-modified nucleosides and deoxyribonucleosides. be done.
  • Each wing segment even more preferably comprises at least one 2'-4' bridged nucleoside.
  • the 2′-4′ bridged nucleosides contained in each wing segment are preferably at least one independently selected from the group consisting of LNA, cEt, ENA, BNA NC , AmNA, GuNA and scpBNA, and more Preferably at least one is independently selected from the group consisting of LNA, AmNA, GuNA and scpBNA.
  • Each wing segment preferably comprises at least one AmNA, more preferably 1-3 AmNAs, and particularly preferably 2 or 3 AmNAs.
  • GuNA contained in each wing segment is particularly preferably GuNA[tBu].
  • AmNA contained in each wing segment is particularly preferably AmNA[Me].
  • the 2′ modified nucleosides contained in each wing segment are preferably 2′-O-Me nucleosides, 2′-O-MOE nucleosides, 2′-O-AP nucleosides, 2′-F nucleosides, 2′-O - at least one independently selected from the group consisting of NMA nucleosides, 2'-O-MCE nucleosides, 2'-DMAECE nucleosides, 2'-MorECE nucleosides, 2'-PyECE nucleosides and 2'-BimECE nucleosides , more preferably at least one independently selected from the group consisting of 2′-O-Me nucleosides, 2′-O-MOE nucleosides, 2′-O-NMA nucleosides and 2′-O-MCE nucleosides , and even more preferably at least one independently selected from 2′-O-Me nucleosides and 2′-O-MCE
  • Each wing segment preferably consists of 1-10 nucleosides independently selected from the group consisting of sugar-modified nucleosides and deoxyribonucleosides and includes at least one 2'-4'-bridged nucleoside. More preferably, it consists of 2 to 6 nucleosides independently selected from the group consisting of 2'-4' bridged nucleosides and deoxyribonucleosides, including at least two 2'-4' bridged nucleosides. More preferably, it consists of 2 to 5 nucleosides independently selected from the group consisting of 2',4'-BNA and deoxyribonucleosides, including at least two 2'-4' bridged nucleosides.
  • each wing segment is more specifically 1 to 10 (preferably 2 to 6, more preferably 2 to 5) independently selected from the group consisting of LNA, AmNA, GuNA, scpBNA and deoxyribonucleosides , more preferably 3) nucleosides, including at least one selected from the group consisting of AmNA, GuNA and scpBNA, more preferably at least one AmNA.
  • each wing segment more preferably consists of 2 to 6 (preferably 2 to 5, more preferably 3) nucleosides independently selected from the group consisting of LNA, AmNA, GuNA and scpBNA, It contains at least one selected from AmNA, GuNA and scpBNA, more preferably at least one AmNA.
  • the 5′ wing segment particularly preferably consists of 2-6 (preferably 2-5, more preferably 3) nucleosides independently selected from the group consisting of AmNA and GuNA, and at least one AmNA including.
  • the 3' wing segment in another embodiment, preferably consists of 2-5 (preferably 3) nucleosides independently selected from the group consisting of AmNA and deoxyribonucleosides and comprises at least one AmNA. More preferably, it contains one deoxyribonucleoside at the 3' end.
  • a preferred LNA contained in each wing segment is ⁇ -D-methyleneoxy BNA.
  • the 5' and 3' wing segments each independently comprise 2, 3, 4, 5, or 6 sugar modified nucleosides and the gap segment comprises 8, 9 , 10, 11, 12, 13, or 14 deoxyribonucleosides.
  • the number of nucleosides of such gapmers can be expressed as (the number of nucleosides in the 5' wing segment - the number of nucleosides in the gap segment - the number of nucleosides in the 3' wing segment), e.g., 5-10-5, 5- 11-4, 4-11-5, 4-12-4, 3-14-3, 6-8-6, 3-12-3, 2-12-2, 2-11-3, 3-11- 2, 3-10-3, 4-10-4, 3-10-4, 4-10-3, 3-9-3, 4-9-4, 3-9-4, 4-9-3, 3-8-3, 3-8-4, 4-8-3 and 4-8-4.
  • the ASO has at least 12, 13, 14, or 15 contiguous nucleosides and the gap segment comprises at least 5, 6, 7, 8, or 9 contiguous nucleosides;
  • the 5′ wing segment and the 3′ wing segment are any of (i) to (iv) below.
  • the 5' and 3' wing segments each independently contain 2, 3, 4, 5 or 6 AmNAs; Particularly preferably, the 5' wing segment and the 3' wing segment each independently contain 3 AmNAs.
  • the 5' wing segment contains 2, 3, 4, 5 or 6 AmNAs and the 3' wing segment contains 2, 3 or 4 AmNAs and 1 or 2 deoxyribonucleosides; Particularly preferably, the 5' wing segment comprises 3 AmNAs and the 3' wing segment comprises 2 AmNAs and 1 deoxyribonucleoside.
  • the 5′ wing segment contains 2, 3, 4, 5 or 6 AmNAs and the 3′ wing segment contains 2, 3 or 4 AmNAs and 1 or 2 scpBNAs.
  • the 5' wing segment comprises 3 AmNAs and the 3' wing segment comprises 2 AmNAs and 1 scpBNA.
  • the 5′ wing segment contains 2, 3, 4, 5 or 6 AmNAs and the 3′ wing segment contains 2, 3 or 4 AmNAs and 1 or 2 LNAs.
  • the 5' wing segment comprises 3 AmNAs and the 3' wing segment comprises 2 AmNAs and 1 LNA.
  • the 5' wing segment comprises 2, 3, 4, 5 or 6 AmNA and the 3' wing segment comprises 1 or 2 AmNA, 1 or 2 scpBNA, 1 or 2 GuNA including.
  • the 5' wing segment comprises 3 AmNAs and the 3' wing segment comprises 1 AmNA, 1 scpBNA and 1 GuNA.
  • the 5' wing segment comprises 2, 3 or 4 AmNA and 1 or 2 Gu and the 3' wing segment comprises 1 or 2 AmNA and 2, 3 or 4 GuNA including. Particularly preferably, the 5' wing segment comprises 2 AmNA and 1 GuNA and the 3' wing segment comprises 1 AmNA and 2 GuNA.
  • the 5′ wing segment and the 3′ wing segment each independently comprise 2, 3 or 4 AmNAs, and 1 or 2 selected from the group consisting of deoxyribonucleosides, scpBNA, GuNA and LNA; include.
  • the arrangement of the sugar modification of the nucleosides constituting the 5′ wing segment in the ASO of the present invention is preferably the group consisting of YYY, YYG, YGY, YGG, GYG, GGY, GGG, GGL, GLG, GLL, LGL and LLL. is more preferably selected from the group consisting of YYY, YGY, and GYG, and particularly preferably YYY.
  • the arrangement of the sugar modification of the nucleosides constituting the 3' wing segment in the ASO of the present invention is preferably YYD, YYS, YYG, YYL, YGG, YGS, YGL, GGS, GYS, GGG, LLL, GLG, LGG and selected from the group consisting of LLG, more preferably selected from the group consisting of YYD, YYS, YYG, YYL, YGG and YGS, more preferably selected from the group consisting of YYD, YYS, YYG and YYL, especially YYS or YYL is preferred.
  • Y is AmNA
  • G is GuNA
  • S is scpBNA
  • L is an LNA
  • D is a deoxyribonucleoside
  • the left side indicates the 5' side and the right side indicates the 3' side.
  • G is GuNA[tBu]
  • Y is AmNA[Me].
  • the internucleoside linkages contained in the 5' wing segment is preferably at least one independently selected from a phosphodiester bond and a modified phosphodiester bond, more preferably at least one independently selected from a phosphodiester bond and a phosphorothioate bond. Seeds.
  • the internucleoside linkages contained in the 5' wing segment preferably comprise at least one phosphorothioate linkage, preferably 50% or more are phosphorothioate linkages, more preferably 60% or more are phosphorothioate linkages, 75 % or more are phosphorothioate linkages, even more preferably 80% or more are phosphorothioate linkages, even more preferably 90% or more are phosphorothioate linkages, and particularly preferably all are phosphorothioate linkages. .
  • the 3' wing segment is similar to the 5' wing segment.
  • the internucleoside linkages contained in each wing segment may be all phosphodiester linkages from the viewpoint of reducing toxicity, but are preferably partially phosphodiester linkages.
  • the 5' terminal internucleoside linkages contained in the 5' wing segment are preferably phosphorothioate linkages.
  • the 3' terminal internucleoside linkages contained in the 3' wing segment are preferably phosphorothioate linkages.
  • the internucleoside bonds other than the 5′ terminal internucleoside bond contained in the 5′ wing segment may be phosphodiester bonds or phosphorothioate bonds, but 50% or more are preferably phosphodiester bonds, More preferably, 60% or more are phosphodiester bonds, more preferably 75% or more are phosphodiester bonds, even more preferably 80% or more are phosphodiester bonds, and 90% or more are phosphodiester bonds. is even more preferred, and all phosphodiester bonds are particularly preferred.
  • the internucleoside bonds other than the 3′ terminal internucleoside bond contained in the 3′ wing segment may be phosphodiester bonds or phosphorothioate bonds, but 50% or more are preferably phosphodiester bonds, More preferably, 60% or more are phosphodiester bonds, more preferably 75% or more are phosphodiester bonds, even more preferably 80% or more are phosphodiester bonds, and 90% or more are phosphodiester bonds. is even more preferred, and all phosphodiester bonds are particularly preferred.
  • the internucleoside bonds located closest to the gap segment is preferably a phosphodiester bond.
  • the 5' terminal internucleoside bond is preferably a phosphorothioate bond
  • the gap segment side internucleoside bond is preferably a phosphodiester bond.
  • the 5' terminal internucleoside linkage is preferably a phosphorothioate linkage
  • the other two internucleoside linkages contained in the 5' wing segment are independent.
  • the 5′ terminal internucleoside bond is preferably a phosphorothioate bond, and the other three internucleoside bonds contained in the 5′ wing segment are independent.
  • a phosphodiester bond or a phosphorothioate bond is preferred, and a phosphodiester bond is particularly preferred.
  • the 3′ terminal internucleoside bond is preferably a phosphorothioate bond
  • the gap segment side internucleoside bond is preferably a phosphodiester bond.
  • the internucleoside bond at the 3′ end is preferably a phosphorothioate bond
  • the other two internucleoside bonds contained in the 3′ wing segment are independent. is preferably a phosphodiester bond or a phosphorothioate bond, and particularly preferably a phosphodiester bond.
  • the internucleoside linkage at the 3' end is preferably a phosphorothioate bond, and the other three internucleoside linkages contained in the 3' wing segment are independent.
  • a phosphodiester bond or a phosphorothioate bond is preferred, and a phosphodiester bond is particularly preferred.
  • the 5' wing segment contains one nucleoside
  • the 5' wing segment contains 0 internucleoside bonds.
  • the 3' wing segment contains one nucleoside
  • the 3' The wing segment contains 0 internucleoside linkages.
  • the 3' end of the 5' wing segment and the 5' end of the gap segment are preferably linked by forming a phosphodiester bond or a modified phosphodiester bond, and the 5' end of the 3' wing segment
  • the 'end and the 3' end of the gap segment are linked by forming a phosphodiester bond or a modified phosphodiester bond.
  • the 3' end of the 5' wing segment and the 5' end of the gap segment are linked by forming a modified phosphodiester bond, and the 5' end of the 3' wing segment and the 3' end of the gap segment are linked.
  • the ends are ligated forming a modified phosphodiester bond.
  • the 3' end of the 5' wing segment and the 5' end of the gap segment are linked by forming a phosphorothioate bond, and the 5' end of the 3' wing segment and the 3' end of the gap segment are linked by phosphorothioate linked by forming a bond.
  • the 3' end of the 5' wing segment and the 5' end of the gap segment are linked by forming a phosphodiester bond, and the 5' end of the 3' wing segment and The 3' ends of the gap segments are linked by forming phosphorothioate bonds.
  • the 3′ end of the 5′ wing segment and the 5′ end of the gap segment are linked by forming a phosphorothioate bond, and the 5′ end of the 3′ wing segment and the 3′ end of the gap segment are linked by phosphorothioate.
  • ends and the 3' end of the gap segment contain an ASO linked by forming a phosphodiester bond.
  • the ASO of the present invention has position numbers 388 to 410, 671 to 688, 726 to 745, 860 to 887, 886 to 957, 1106 to 1130, 1157 of the nucleobase of the FGFR3 mRNA represented by SEQ ID NO: 1.
  • the ASO of the present invention has position numbers 388 to 410, 671 to 686, 726 to 743, 860 to 887, 886 to 956, 1106 to 1130, 1157 of the nucleobase of the FGFR3 mRNA represented by SEQ ID NO: 1.
  • nucleobases represented by Complementary to at least 12 contiguous nucleobases within the nucleobase sequence selected from the group consisting of sequences.
  • the ASO of the present invention preferably has nucleobase positions 388-410, 671-688, 726-745, 860-887, 886-915, 924-957, 1106- 1130,1157-1174,1195-1210,1216-1231,1238-1256,1276-1294,1574-1588,1622-1647,1648-1662,1730-1750,1750-1771,1779-1795,1822-1836, 1850-1870, 1991-2005, 2017-2058, 2083-2112, 2189-2203, 2250-2264, 2528-2542, 2711-2726, 2782-2804, 2968-2987, 3203-3217, 3558-3572, 3656- selected from the group consisting of nucleic acid base sequences represented by is complementary to at least 12 contiguous nucleobases within the nucleobase sequence of
  • the ASO of the present invention is at position numbers 388-410, 671-686, 726-743, 860-887, 886-915, 929-956, 1106 of the nucleobase of the FGFR3 mRNA represented by SEQ ID NO: 1.
  • the ASO of the present invention has position numbers 388-402, 389-403, 390-404, 391-405, 392-406, 393-407, 394 of the nucleobase of the FGFR3 mRNA represented by SEQ ID NO: 1.
  • the ASO of the present invention is more preferably nucleobase positions 673-687, 674-688, 731-745, 924-938, 925-939, 926- 940, 927-941, 928-942, 943-957, 1195-1209, 1279-1293, 1280-1294, 1730-1744, 1731-1745, 1732-1746, 1733-1747, 1736-1750, 1856-1870, complementary to at least 12 contiguous nucleobases within the nucleobase sequence shown in 2020-2034, 2021-2035, 2022-2036, 2023-2037, 2029-2043 or 2972-2986.
  • the ASO of the present invention has the nucleobase position numbers 4040-4054, 4041-4055, 4042-4056, 4043-4057 of the FGFR3 mRNA represented by SEQ ID NO: 1, is complementary to at least 12 contiguous nucleobases within the nucleobase sequence designated 4044-4058, 4045-4059, 4046-4060, 4047-4061 or 4048-4062.
  • the ASO of the present invention has the nucleobase position numbers 1734-1748, 1735-1749, 1750-1764, 1751-1765, 1752-1766, 1752-1768, 1752-1770 of the FGFR3 mRNA represented by SEQ ID NO: 1. , 1753-1767, 1753-1769, 1754-1768, 1755-1769, 1756-1770, or 1757-1771.
  • the ASO of the present invention has the nucleobase position numbers 390-404, 391-405, 394-408, 396-410, 869-883, 1110-1124 of the FGFR3 mRNA represented by SEQ ID NO: 1, 1111-1125, 1115-1129, 1116-1130, 1238-1252, 1242-1256, 1622-1636, 1623-1637, 1627-1641, 1630-1644, 1753-1767, 1754-1768, 1755-1769, 2042- 2056, 2785-2799, 2789-2803, 3823-3837, 3924-3938, 3925-3939, 4007-4021, 4008-4022, 4009-4023, 4043-4057, 4044-4058, 4048-4062, 4098-4112, 4099-4113, 4100-4114, 4101-4115, 4102-4116, or 4250-4264 is complementary to at least 12 contiguous nucleobases within the nucleobase sequence shown in
  • the ASOs of the present invention are particularly preferably complementary to at least 12 contiguous nucleobases within the nucleobase sequence shown in nucleobase positions 1753-1767 of the FGFR3 mRNA represented by SEQ ID NO:1.
  • the ASOs of the present invention are particularly preferably complementary to at least 12 contiguous nucleobases within the nucleobase sequence shown in nucleobase positions 1754-1768 of the FGFR3 mRNA represented by SEQ ID NO:1.
  • the ASOs of the present invention are particularly preferably complementary to at least 12 contiguous nucleobases within the nucleobase sequence shown in nucleobase positions 1755-1769 of the FGFR3 mRNA represented by SEQ ID NO:1.
  • the ASOs of the present invention are particularly preferably complementary to at least 12 contiguous nucleobases within the nucleobase sequence represented by nucleobase positions 4043-4057 of the FGFR3 mRNA represented by SEQ ID NO:1.
  • the ASOs of the present invention are particularly preferably complementary to at least 12 contiguous nucleobases within the nucleobase sequence shown in nucleobase positions 4044-4058 of the FGFR3 mRNA represented by SEQ ID NO:1.
  • the ASOs of the present invention are particularly preferably complementary to at least 12 contiguous nucleobases within the nucleobase sequence represented by nucleobase positions 4048-4062 of the FGFR3 mRNA represented by SEQ ID NO:1.
  • An ASO complementary to at least 12 contiguous nucleobases in the nucleobase sequence is not described in WO03/023004 and WO2014/080004.
  • An ASO complementary to at least 12 contiguous nucleobases within said nucleobase sequence is preferably complementary to at least 13, 14, 15 contiguous nucleobases within said nucleobase sequence.
  • the ASO of the present invention preferably has FGFR3 mRNA nucleobase position numbers 675-690, 874-899, 944-967, 1205-1229, 1299-1314, 1748-1763, 2693- It is complementary to at least a portion within the nucleobase sequence shown at 2707, 4048-4071, or 4251-4270.
  • the ASOs of the present invention are preferably nucleobase position numbers 874-899, 948-962, 1213-1229, 1299-1314, 1748-1763, 2693-2707, 4048-4071 of the FGFR3 mRNA represented by SEQ ID NO: 1. , or at least a portion within the nucleobase sequence shown in 4251-4270.
  • the ASOs of the present invention are more preferably ⁇ 898, 885 ⁇ 899, 948 ⁇ 962, 1213 ⁇ 1227, 1214 ⁇ 1228, 1215 ⁇ 1229, 1299 ⁇ 1313, 1300 ⁇ 1314, 1748 ⁇ 1762, 1749 ⁇ 1763, 2693 ⁇ 2707, 4048 ⁇ 4064, 4048 ⁇ 4066 ,4049-4063,4050-4064,4051-4065,4052-4066,4052-4067,4052-4068,4053-4067,4054-4068,4055-4069,4056-4070,4057-4071,4251-4265,4252 -4266, 4253-4267, 4254-4268, 4255-4269 or 4256-4270.
  • the ASO of the present invention is more preferably nucleobase positions 675-689, 676-690, 944-958, 945-959, 946-960, 947- 961, 949-963, 950-964, 951-965, 952-966, 953-967, 1205-1219, 1211-1225, or 1212-1226.
  • the ASOs of the present invention are more preferably -4067, 4052-4068, 4053-4067, 4054-4068, 4055-4069, 4056-4070, or 4057-4071.
  • the ASO of the present invention is complementary to at least a portion within the nucleobase sequence represented by nucleobase positions 1748-1762 or 1749-1763 of the FGFR3 mRNA represented by SEQ ID NO:1.
  • the ASO of the present invention has nucleobase position numbers 875-889, 1299-1313, 4051-4065, 4053-4067, 4054-4068, 4055-4069, 4056 of the nucleobase of FGFR3 mRNA represented by SEQ ID NO: 1. It is complementary to at least a portion within the nucleobase sequence shown by ⁇ 4070 or 4255-4269.
  • the ASO of the present invention is more preferably at positions 675 to 689, 947 to 961, 951 to 965, 952 to 966, or 953 to 967 of the nucleobase of the FGFR3 mRNA represented by SEQ ID NO: 1. It is complementary to at least a portion within the indicated nucleobase sequence.
  • the ASO of the present invention is particularly preferably complementary to at least part of the nucleobase sequence represented by position numbers 4051-4065 of the FGFR3 mRNA represented by SEQ ID NO:1.
  • the ASO of the present invention is particularly preferably complementary to at least part of the nucleobase sequence represented by position numbers 4053-4067 of the FGFR3 mRNA represented by SEQ ID NO:1.
  • the ASO of the present invention is particularly preferably complementary to at least part of the nucleobase sequence represented by position numbers 4054-4068 of the FGFR3 mRNA represented by SEQ ID NO:1.
  • the ASO of the present invention is particularly preferably complementary to at least part of the nucleobase sequence represented by position numbers 4055-4069 of the FGFR3 mRNA represented by SEQ ID NO:1.
  • the ASO of the present invention is particularly preferably complementary to at least part of the nucleobase sequence represented by position numbers 4056-4070 of the FGFR3 mRNA represented by SEQ ID NO:1.
  • the ASO of the present invention is particularly preferably complementary to at least part of the nucleobase sequence represented by position numbers 947-961 of the FGFR3 mRNA represented by SEQ ID NO:1.
  • ASOs complementary to at least a portion within said nucleobase sequence and comprising at least one 2'-4'-bridged nucleoside are disclosed in WO 03/023004 and WO 2014/080004 not listed in the number.
  • An ASO complementary to at least a portion within said nucleobase sequence is preferably complementary to at least 12, 13, 14, 15 contiguous nucleobases within said nucleobase sequence.
  • Complementarity between the ASO of the present invention and the relevant portion of FGFR3 mRNA represented by SEQ ID NO: 1 is at least 90% or more, still more preferably 95%, 96%, 97% , 98%, 99% or more.
  • the ASO of the present invention preferably consists of 12-30 nucleosides complementary to at least a portion of each of the target regions of FGFR3 mRNA represented by SEQ ID NO:1. More preferably 12 to 20, more preferably 14 to 20, still more preferably 15 to 19, particularly preferably 15 nucleosides.
  • the nucleobase sequence possessed by the ASO of the present invention preferably comprises at least 12, more preferably 13, 14 or 15 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOS:2-225.
  • the nucleobase sequence possessed by the ASO of the present invention more preferably includes any of the nucleobase sequences of SEQ ID NOS:2-225.
  • the nucleobase sequence of the ASO of the present invention is preferably at least 12, more preferably 13, 14 or 15 contiguous nucleic acids of any one of nucleobase sequence groups A (A1 and A2). It has a nucleobase sequence containing bases, more preferably contains any one nucleobase sequence of nucleobase sequence group A (A1 and A2), still more preferably any one of nucleobase sequence group A (A1 and A2) or one nucleobase sequence.
  • ASOs comprising at least 12 contiguous nucleobases of the nucleobase sequence of any one of nucleobase sequence group A (A1 and A2) are described in WO 03/023004 and WO 2014/080004. do not have.
  • the nucleobase sequence of the ASO of the present invention is more preferably at least 12, more preferably 13, 14 or 15 contiguous nucleobases of any one of the nucleobase sequence group B (B1). More preferably any one nucleobase sequence of nucleobase sequence group B (B1), still more preferably any one nucleobase of nucleobase sequence group B (B1) has an array.
  • the nucleobase sequence possessed by the ASO of the present invention is, as another aspect, more preferably at least 12, 13, Having a nucleobase sequence containing 14 or 15 contiguous nucleobases, more preferably containing any one nucleobase sequence of nucleobase sequence group C (C1 and C2), still more preferably nucleobase sequence group C It has any one nucleic acid base sequence of (C1 and C2).
  • An ASO comprising at least 12 contiguous nucleobases of any one nucleobase sequence of nucleobase sequence group C (C1 and C2) and comprising at least one 2'-4'-bridged nucleoside is WO 03 /023004 and WO2014/080004.
  • the nucleobase sequence of the ASO of the present invention is more preferably at least 12, more preferably 13, 14 or 15 contiguous nucleobase sequences of any one of the nucleobase sequence groups D (D1 and D2). It has a nucleobase sequence containing a nucleobase, more preferably contains any one nucleobase sequence of nucleobase sequence group D (D1 and D2), still more preferably of nucleobase sequence group D (D1 and D2) It has any one nucleic acid base sequence.
  • the ASO of the present invention consists of 12 to 30, more preferably 12 to 20, still more preferably 14 to 20, still more preferably 15 to 19, particularly preferably 15 nucleosides.
  • Nucleic acid base sequence group A1 SEQ ID NO: 2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,21,22,23,27,28,29, 30, 31, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 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, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 92, 94, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 115, 116, 117, 118,
  • nucleic acid base sequence group A1 preferred SEQ ID NOs: 35, 36, 37, 108, 109, 110, 111, 155, 156, 157, 158, 159, 197, 198, 199, 200, 201, 219, 220 , and 221.
  • Nucleic acid base sequence group A2 from SEQ. the group.
  • Nucleic acid base sequence group B1 SEQ. The group consisting of 182, 183, 184, 185, 190, 200, 201, 208, 209, 210, 211, 212 and 214. Of the nucleic acid base sequence group B1, particularly preferred sequence numbers are any of 111, 157, 158, 159, 200 and 201.
  • Nucleic acid base sequence group C1 SEQ. The group consisting of 204, 205, 206, 207, 215, 216, 217, 218, 222, 223, 224 and 225. Of the nucleic acid base sequence group C1, preferred sequence numbers are any of 112, 113, 114, 153, 154, 202, 203, 204, 205, 206, 207, 222, 223, 224 and 225.
  • Nucleic acid base sequence group C2 The group consisting of SEQ.
  • Nucleic acid base sequence group D1 The group consisting of SEQ ID NOS: 32, 139, 202, 203, 204, 205, 206, and 218. Of the nucleic acid base sequence group D1, particularly preferred sequence numbers are any of 202, 203, 204, 205 and 206.
  • Nucleic acid base sequence group D2 The group consisting of SEQ ID NOS: 228, 240, 243, 244 and 245.
  • each wing segment preferably comprises three 2' -4' bridged nucleosides, more preferably at least 1 AmNA, more preferably 2 or 3 AmNAs. More preferably, the 3' wing segment has 3 AmNAs and the 5' wing segment has 2 AmNAs and one selected from the group consisting of scpBNA, LNA, GuNA and deoxyribonucleosides. Even more preferably, the 3' wing segment has 3 AmNAs and the 5' wing segment has 2 AmNAs and 1 scpBNA or 1 LNA. Particularly preferably, the 3' wing segment has 3 AmNAs and the 5' wing segment has 2 AmNAs and 1 scpBNA. AmNA is particularly preferably AmNA[Me] and GuNA is particularly preferably GuNA[tBu].
  • nucleobase sequence groups A, B, C and D particularly preferred SEQ ID NOs are sequences 111, 204 and 240.
  • the arrangement of sugar modification in the above sequence is selected from the following combinations.
  • the 5' wing segment is YYY, the 3' wing segment is YYS, and the gap segment is DDDDDDDD.
  • the 5' wing segment is YYY, the 3' wing segment is YYL and the gap segment is DDDDDDDD.
  • the 5' wing segment is YYY, the 3' wing segment is YYS, and the gap segment is ZZDDDDDD or ZZZDDDDDD.
  • the 5' wing segment is YYY
  • the 3' wing segment is YYL
  • the gap segment is ZZZDDDDDD.
  • the arrangement of the sugar modification in the sequence is more preferably (i) or (iii) for SEQ ID NO: 111, (ii) or (iv) for SEQ ID NO: 204, and SEQ ID NO: 240.
  • the left side indicates the 5' side and the right side indicates the 3' side
  • Y is AmNA
  • S is scpBNA
  • L is an LNA
  • D is a deoxyribonucleoside
  • Z is a 5'-CP nucleoside.
  • Y is AmNA[Me].
  • a functional molecule may be bound directly or indirectly to the ASO of the present invention.
  • the binding between the functional molecule and the ASO may be direct or indirect via another substance, but it is preferable that the oligonucleotide and the functional molecule are bound by a covalent bond, an ionic bond or a hydrogen bond. . From the viewpoint of high stability of the bond, it is more preferable to bond directly by covalent bond or to bond by covalent bond via a linker (linking group).
  • said functional molecule When said functional molecule is covalently bound to ASO, it is preferred that said functional molecule is bound directly or indirectly to the 3' or 5' end of the ASO molecule.
  • the linkage between the linker or functional molecule and the terminal nucleoside of the ASO molecule is selected according to the functional molecule.
  • the linker or functional molecule and the terminal nucleoside of the ASO molecule are preferably linked by a phosphodiester bond or a modified phosphodiester bond.
  • the linker or functional molecule may be directly linked to the 3′-position oxygen atom of the 3′-terminal nucleoside or the 5′-position oxygen atom of the 5′-terminal nucleoside of the ASO molecule.
  • the structure of the "functional molecule” is not particularly limited, and the desired function is imparted to the ASO by its binding. Desired functions include labeling functions, purification functions and delivery functions to target sites. Examples of molecules that impart a labeling function include compounds such as fluorescent proteins and luciferase. Examples of molecules that confer a purification function include compounds such as biotin, avidin, His-tag peptides, GST-tag peptides, FLAG-tag peptides.
  • FGFR3 gene is preferably bound to a molecule that has the function of delivering the ASO to the target site.
  • a sugar derivative that interacts with the asialoglycoprotein receptor may be used as a functional molecule capable of delivering the ASO of the present invention with high specificity to the liver.
  • the "asialoglycoprotein receptor" exists on the surface of liver cells, and has the action of recognizing the galactose residue of asialoglycoprotein and taking that molecule into the cell and degrading it.
  • the "sugar derivative that interacts with the asialoglycoprotein receptor” is preferably a compound that has a structure similar to that of a galactose residue and is taken up into cells by interacting with the asialoglycoprotein receptor. acetylgalactosamine) derivatives, galactose derivatives and lactose derivatives.
  • Functional molecules that can efficiently deliver ASO to the organ with high specificity by interacting with various proteins on the cell surface of each organ include receptor ligands, antibodies, and peptides or proteins that are fragments thereof. mentioned.
  • the linker through which the functional molecule and the ASO are bound can exhibit the function of the functional molecule as the ASO molecule, it is particularly preferable that the linker stably binds the functional molecule and the oligonucleotide.
  • the linker include a group derived from an oligonucleotide having 1 to 20 nucleosides, a group derived from a polypeptide having 1 to 20 amino acids, an alkylene having 1 to 20 carbons and an alkylene having 2 to 20 carbons. and the like.
  • the group derived from an oligonucleotide having 1 to 20 nucleosides is a hydrogen atom or the like from the 3' end and 5' end of an oligonucleotide having 1 to 20 nucleosides (a nucleoside when the number of nucleosides is 1). is a divalent group from which is removed.
  • groups derived from oligonucleotides having 1 to 20 nucleosides see, for example, International Publication No. 2017/053995.
  • International Publication No. 2017/053995 describes, for example, a 3-base linker having a TCA motif, a 1-5 base linker having no TCA motif, and the like.
  • the group derived from a polypeptide having 1 to 20 amino acids is selected from polypeptides having 1 to 20 amino acids (amino acids when the number of amino acids is 1), hydroxy, hydrogen atoms, amino, etc. 2 It is a divalent group with one group removed.
  • the ASO-containing compounds of the present invention may contain prodrug moieties.
  • a prodrug is a derivative of a pharmaceutical compound having groups that are chemically or metabolically degradable, and which, upon solvolysis or in vivo degradation under physiological conditions, render the pharmacologically active pharmaceutical compound is a compound that induces Methods for selecting and manufacturing suitable prodrug derivatives are described, for example, in Design of Prodrugs (Elsevier, Amsterdam, 1985).
  • a prodrug moiety such as an acyloxy derivative prepared by reacting the hydroxy group of ASO (or a compound containing ASO) with a suitable acyl halide, a suitable acid anhydride or a suitable halogenated alkyloxycarbonyl compound.
  • the prodrugs of the ASO-containing compounds of the present invention also include complexes and the like formed by hybridizing two or more oligonucleotides.
  • Such preferred structures (prodrugs) include oligonucleotides containing ribonucleosides (e.g., RNA), oligonucleotides containing peptide nucleic acids (PNA), or deoxyribonucleosides complementary to ASO (or compounds containing ASO).
  • a double-stranded oligonucleotide comprising an oligonucleotide (e.g., DNA) comprising , oligonucleotides in which RNA complementary to ASO is bound by a linker (eg, International Publication No. 2017/131124, International Publication No. 2018/143475), and the like.
  • the linkers may be oligonucleotide linkers or linkers comprising non-nucleoside structures. Oligonucleotides in which RNA complementary to ASO is directly linked are also included (WO2019/022196).
  • the compounds of the present invention include compounds in which two identical or different ASOs are linked.
  • structures that connect two ASOs see, for example, WO2017/131124 and WO2018/143475.
  • Compounds containing ASO include those that exist via their tautomerism and geometric isomerism, as well as those that exist as mixtures thereof or mixtures of their respective isomers.
  • an asymmetric center exists, or when an asymmetric center is generated as a result of isomerization, it also includes those existing as respective optical isomers and mixtures of arbitrary ratios.
  • diastereomers due to each optical isomerism also exist.
  • the present invention also includes all these types in any proportion. Optically active forms can also be obtained by well-known methods for this purpose.
  • oligonucleotides and phosphorus atom sterically controlled oligonucleotides Any form of conformationally uncontrolled oligonucleotide of is included within the scope of the present invention.
  • the ASO-containing compounds of the present invention, their prodrugs, or pharmacologically acceptable salts thereof can exist as any crystal form and as any hydrate depending on the production conditions, These crystalline forms and hydrates and mixtures thereof are also included within the scope of the present invention. It may also exist as solvates with organic solvents such as acetone, ethanol, 1-propanol, 2-propanol, etc., and all of these forms are included within the scope of the present invention.
  • the ASO-containing compounds of the present invention can be converted into pharmacologically acceptable salts or liberated from the salts formed, if desired.
  • pharmacologically acceptable salts of compounds containing ASO include alkali metals (lithium, sodium, potassium, etc.), alkaline earth metals (calcium, etc.), magnesium, ammonium, organic bases (triethylamine, trimethylamine, etc.), Amino acids (glycine, lysine, glutamic acid, etc.), inorganic acids (hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, etc.) or organic acids (acetic acid, citric acid, maleic acid, fumaric acid, tartaric acid, benzenesulfonic acid, methanesulfonic acid) , p-toluenesulfonic acid, etc.).
  • a pharmacologically acceptable salt is particularly preferably the sodium salt.
  • a compound containing ASO of the present invention can be prepared by a person skilled in the art by appropriately selecting a known method.
  • a person skilled in the art can design the nucleoside sequence of ASO based on the nucleoside sequence information of the target RNA, and use a commercially available automatic nucleic acid synthesizer (manufactured by Applied Biosystems, Beckman, Genedesign, etc.). can be synthesized by Alternatively, it can be synthesized by a reaction using an enzyme.
  • the enzymes include, but are not limited to, polymerases, ligases, restriction enzymes, and the like. That is, the method for producing an ASO-containing compound according to this embodiment can include the step of extending the nucleoside chain at the 3' end or the 5' end.
  • a compound containing ASO can be prepared by purifying the resulting oligonucleotide by reverse-phase column chromatography or the like.
  • the ASO-containing compound of the present invention or a pharmacologically acceptable salt thereof can effectively inhibit the expression of the FGFR3 gene.
  • Diseases that can be treated, prevented, or ameliorated by a nucleic acid drug that utilizes a compound containing an ASO of the present invention or a pharmacologically acceptable salt thereof are not particularly limited as long as the FGFR3 gene expression inhibitory action is effective.
  • achondroplasia including severe achondroplasia with developmental delay and acanthosis nigricans
  • achondroplasia tanatophoric osteodysplasia (types I and II)
  • Crouzon's disease accelerated fracture healing Among them, achondroplasia is exemplified.
  • FGFR3 gene is associated with cancer, and can be applied to cancer diseases.
  • Applicable cancer diseases include colon cancer, bladder cancer, bone cancer, lung cancer, cervical cancer, breast cancer, skin cancer, testicular cancer, thyroid cancer, hepatocellular carcinoma, melanoma, prostate cancer, squamous cell carcinoma, and the like.
  • the present invention can provide, for example, a composition for inhibiting the expression of the FGFR3 gene by antisense effect, which contains the compound containing the ASO as an active ingredient.
  • compounds comprising the ASOs of the present invention are effective in reducing the expression of the FGFR3 gene, such as achondroplasia, achondroplasia, tanatophoric osteodysplasia (types I and II), Crouzon's disease, accelerated fracture healing, and cancer.
  • Pharmaceutical compositions can also be provided for treating, preventing, and/or ameliorating diseases for which inhibition is beneficial.
  • a pharmaceutical composition containing the ASO-containing compound of the present invention or a pharmacologically acceptable salt thereof can be formulated by a known pharmaceutical method.
  • Turbidities, elixirs, emulsions, liniments, ointments, plasters, poultices, transdermal preparations, lotions, inhalants, aerosols, suppositories, etc. can be administered enterally (orally, etc.) or non- It can be used enterally.
  • pharmacologically acceptable carriers specifically sterile water, physiological saline, vegetable oils, solvents, bases, emulsifiers, suspending agents, surfactants, pH adjusters , stabilizers, flavoring agents, fragrances, excipients, vehicles, preservatives, binders, diluents, tonicity agents, soothing agents, bulking agents, disintegrants, buffers, coating agents, lubricants, Coloring agents, sweetening agents, thickening agents, flavoring agents, solubilizing agents, other additives, and the like can be appropriately combined.
  • the mode of administration of the composition containing the ASO-containing compound of the present invention or a pharmacologically acceptable salt thereof is not particularly limited, and includes enteral (oral, etc.) or parenteral administration. More preferably, intravenous administration, intraarterial administration, intraarticular administration, intraperitoneal administration, subcutaneous administration, intradermal administration, intratracheal administration, rectal administration, intramuscular administration, intrathecal administration, intracerebroventricular administration, intranasal administration and intravitreal administration and administration by infusion, more preferably intravenous administration, intraarticular administration and subcutaneous administration, particularly preferably subcutaneous administration.
  • Preparations for subcutaneous administration, intra-articular administration, intravenous administration, etc., containing the ASO-containing compound of the present invention or a pharmacologically acceptable salt thereof can be manufactured by known conventional methods.
  • a compound containing ASO of the present invention or a pharmacologically acceptable salt thereof and additives such as a buffer and a tonicity agent are completely dissolved in water for injection, and then filtered and sterilized.
  • a pre-filled syringe formulation is obtained by filling a sterilized syringe with the preparation solution, a preparation for injection is obtained by filling a sterilized vial, and a ready-to-use preparation is obtained by filling a sterilized vial and freeze-drying it.
  • Final sterilization such as autoclaving or gamma irradiation may be performed instead of filtration sterilization.
  • a variety of mammalian diseases can be treated, prevented and/or ameliorated by a composition containing the ASO-containing compound of the present invention or a pharmacologically acceptable salt thereof.
  • a composition containing the ASO-containing compound of the present invention or a pharmacologically acceptable salt thereof For example, but not limited to, humans, cows, sheep, goats, horses, dogs, cats, guinea pigs, rats, rabbits, chimpanzees or other bovines. ), ovine, equine, canine, feline, mouse and other rodent species.
  • the mammal is particularly preferably human.
  • the dosage or intake is preferably an effective amount.
  • Effective amount means an amount of a compound sufficient to produce the desired pharmacological effect in an individual in need of the drug, and to treat, prevent and/or ameliorate the disease, age, weight, symptoms, health status of the individual. , varies depending on the formulation of the composition, etc., and is selected as appropriate.
  • the dosage or intake is preferably 0.0001 mg/kg/day to 100 mg/kg/day in terms of ASO.
  • a method for regulating FGFR3 gene expression comprising the step of contacting a cell with a compound containing an ASO of the present invention or a pharmacologically acceptable salt thereof.
  • a method for regulating FGFR3 gene expression in a mammal comprising the step of administering a pharmaceutical composition comprising a compound comprising an ASO of the present invention or a pharmacologically acceptable salt thereof to said mammal.
  • the internucleoside bond between the two nucleosides is a phospho It is a diester bond.
  • the internucleoside linkage between A and G is a phosphodiester bond
  • the internucleoside linkages between are phosphodiester linkages.
  • Antisense oligonucleotides (compounds represented by chemical structures corresponding to compound numbers) listed in Tables 1 to 8 were prepared using an automatic nucleic acid synthesizer (nS-8 type, manufactured by Gene Design Co., Ltd.). . Note that the title lines of Tables 2 to 8 are the same as those of Table 1, so they are omitted.
  • SEQ1 START means "SEQ ID NO: 1 start site”
  • SEQ ID NO: 1 start site the most 5' site targeted by the antisense oligonucleotide in the human FGFR3 mRNA sequence (SEQ ID NO: 1).
  • the position numbers of the side nucleosides are indicated.
  • SEQ1 END means "SEQ ID NO: 1 end site” and indicates the position number of the 3'-most nucleoside targeted by the antisense oligonucleotide in the human FGFR3 mRNA sequence (SEQ ID NO: 1).
  • SEQ No indicates the sequence number
  • BASE SEQUENCE indicates the nucleic acid base sequence of the antisense oligonucleotide. Note that the title lines of Tables 10 to 16 are the same as those of Table 9, so they are omitted.
  • ISIS #125189 of US2004/0048824 was prepared using an automatic nucleic acid synthesizer (nS-8 type, manufactured by Genedesign).
  • the compound is herein LX-A4921.
  • the nucleobase sequence of LX-A4921 is represented by ATGCTGCCAAAACTTGTTCTC (SEQ ID NO:276).
  • the FGFR3 gene expression level was measured by quantitative real-time PCR using TaqMan (registered trademark) Gene Expression Assays (Thermo Fisher Scientific).
  • the mRNA level of the housekeeping gene peptidylprolyl isomerase A [PPIA] was also quantified at the same time, and the FGFR3 mRNA level relative to the PPIA mRNA level was evaluated as the FGFR3 expression level.
  • Results are expressed in Tables 17-19 as percent expression of FGFR3 relative to untreated control cells.
  • Table 17 shows the data under the condition that the overnight starvation culture was performed, and Tables 18 and 19 show the data under the condition that the overnight starvation culture was not performed.
  • PrimeScript registered trademark
  • RT Master Mix Perfect Real Time
  • FGFR1, FGFR3, and FGFR4 gene expression levels were measured by quantitative real-time PCR using TaqMan (registered trademark) Gene Expression Assays (Thermo Fisher Scientific).
  • the amount of PPIA mRNA was also quantified at the same time, and the amount of FGFR1, FGFR3, and FGFR4 mRNA relative to the amount of PPIA mRNA was evaluated as the expression level of FGFR1, FGFR3, and FGFR4.
  • antisense oligonucleotides with high FGFR3 inhibitory activity and low FGFR1 and FGFR4 expression inhibitory activity are desirable.
  • LX-A3531, 4393, 4989, 5534, 5536, 6798, 6799, 6908 and LX-A4921 were evaluated by the method of Evaluation Example 3. These compounds inhibited FGFR3 in a concentration-dependent manner. LX-A3531, 4393, 5534, 5536, 6798, 6799 and 6908 showed higher activity than LX-A4921. The FGFR4 mRNA expression level of these compounds showed a survival rate of 75% or more relative to untreated control cells at any concentration, indicating no concentration-dependent inhibition. FGFR3 was shown to be highly selective over FGFR4.
  • the FGFR1 mRNA expression levels of LX-A3531, 4393, 4989, 6798, 6799, and 6908 showed a residual rate of 75% or more relative to untreated control cells at any concentration, indicating concentration-dependent inhibition. I didn't.
  • the expression levels of FGFR1 and FGFR3 relative to untreated control cells when LX-A4393, 5534, 5536, 6798, 6799 and LX-A4921 were added are presented in FIG.
  • the selectivity of FGFR3 over FGFR1 was shown to be higher for LX-A3531, 4393, 4989, 5534, 5536, 6798, 6799, 6908 than for LX-A4921.
  • FGFR2 and FGFR3 gene expression levels were measured by quantitative real-time PCR using TaqMan (registered trademark) Gene Expression Assays (Thermo Fisher Scientific).
  • TaqMan registered trademark
  • Gene Expression Assays Thermo Fisher Scientific
  • the amount of PPIA mRNA which is a housekeeping gene, was also quantified at the same time, and the amount of FGFR2 and FGFR3 mRNA relative to the amount of PPIA mRNA was evaluated as the expression level of FGFR2 and FGFR3.
  • antisense oligonucleotides with high FGFR3 inhibitory activity and low FGFR2 expression inhibitory activity are desirable.
  • LX-A3531, 4393, 4989, 5534, 5536, 6798, 6799, 6908 and LX-A4921 were evaluated by the method of Evaluation Example 4. These compounds inhibited FGFR3 in a concentration-dependent manner. When these compounds were added, the FGFR2 mRNA expression level remained at 75% or more of the untreated control cells at any concentration, showing no concentration-dependent inhibition. FGFR3 was shown to be highly selective over FGFR2.
  • RNA from cartilage tissue containing growth plates was extracted using ISOGEN II (NIPPON GENE CO., LTD.) according to the enclosed protocol, RNA was isolated using RNeasy Mini Kit (QIAGEN), and PrimeScript cDNA was prepared by reverse transcription using ® RT Master Mix (Perfect Real Time) (Takara Bio). Using the prepared cDNA, the FGFR3 gene expression level was measured by quantitative real-time PCR using TaqMan (registered trademark) Gene Expression Assays (Thermo Fisher Scientific).
  • RNA from cartilage tissue containing growth plates was extracted using ISOGEN II (NIPPON GENE CO., LTD.) according to the enclosed protocol, RNA was isolated using RNeasy Mini Kit (QIAGEN), and PrimeScript cDNA was prepared by reverse transcription using ® RT Master Mix (Perfect Real Time) (Takara Bio). Using the prepared cDNA, the FGFR3 gene expression level was measured by quantitative real-time PCR using TaqMan (registered trademark) Gene Expression Assays (Thermo Fisher Scientific).
  • IL-6 concentration contained in the culture supernatant was measured by ELISA (Enzyme-Linked Immuno Sorbent Assay) method, and the amount of IL-6 mRNA in cultured cells was measured by TaqMan. (registered trademark) Gene Expression Assays (Thermo Fisher Scientific).
  • ELISA Enzyme-Linked Immuno Sorbent Assay
  • TaqMan registered trademark
  • Gene Expression Assays Thermo Fisher Scientific
  • the antisense oligonucleotides of the present invention inhibit the expression of the FGFR3 gene, they are useful for the treatment, prevention, and/or amelioration of diseases for which inhibition of FGFR3 gene expression is effective, particularly achondroplasia.

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US20040048824A1 (en) * 2001-09-10 2004-03-11 Monia Brett P. Antisense modulation of fibroblast growth factor receptor 3 expression
JP2016501520A (ja) * 2012-11-26 2016-01-21 ロシュ・イノベーション・センター・コペンハーゲン・アクティーゼルスカブＲｏｃｈｅ Ｉｎｎｏｖａｔｉｏｎ Ｃｅｎｔｅｒ Ｃｏｐｅｎｈａｇｅｎ Ａ／Ｓ Ｆｇｆｒ３の発現の調節のための組成物及び方法
WO2019182037A1 (ja) * 2018-03-20 2019-09-26 国立大学法人東京工業大学 毒性が低減されたアンチセンスオリゴヌクレオチド
WO2020152869A1 (ja) * 2019-01-25 2020-07-30 杏林製薬株式会社 線維症治療剤
WO2020158910A1 (ja) * 2019-02-01 2020-08-06 国立大学法人大阪大学 ５'位修飾ヌクレオシドおよびそれを用いたヌクレオチド

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US20040048824A1 (en) * 2001-09-10 2004-03-11 Monia Brett P. Antisense modulation of fibroblast growth factor receptor 3 expression
JP2016501520A (ja) * 2012-11-26 2016-01-21 ロシュ・イノベーション・センター・コペンハーゲン・アクティーゼルスカブＲｏｃｈｅ Ｉｎｎｏｖａｔｉｏｎ Ｃｅｎｔｅｒ Ｃｏｐｅｎｈａｇｅｎ Ａ／Ｓ Ｆｇｆｒ３の発現の調節のための組成物及び方法
WO2019182037A1 (ja) * 2018-03-20 2019-09-26 国立大学法人東京工業大学 毒性が低減されたアンチセンスオリゴヌクレオチド
WO2020152869A1 (ja) * 2019-01-25 2020-07-30 杏林製薬株式会社 線維症治療剤
WO2020158910A1 (ja) * 2019-02-01 2020-08-06 国立大学法人大阪大学 ５'位修飾ヌクレオシドおよびそれを用いたヌクレオチド

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024071099A1 (ja) * 2022-09-29 2024-04-04 国立大学法人東京医科歯科大学 ５'－シクロプロピレン修飾を含む核酸分子

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