JP7656784B2 - Novel compounds and anti-coronavirus agents - Google Patents
Novel compounds and anti-coronavirus agents Download PDFInfo
- Publication number
- JP7656784B2 JP7656784B2 JP2020195723A JP2020195723A JP7656784B2 JP 7656784 B2 JP7656784 B2 JP 7656784B2 JP 2020195723 A JP2020195723 A JP 2020195723A JP 2020195723 A JP2020195723 A JP 2020195723A JP 7656784 B2 JP7656784 B2 JP 7656784B2
- Authority
- JP
- Japan
- Prior art keywords
- coronavirus
- agent
- active ingredient
- agent according
- concentration
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 150000001875 compounds Chemical class 0.000 title claims description 38
- 241000711573 Coronaviridae Species 0.000 claims description 52
- 239000004480 active ingredient Substances 0.000 claims description 20
- 230000000840 anti-viral effect Effects 0.000 claims description 16
- 241001678559 COVID-19 virus Species 0.000 claims description 15
- 241000004175 Coronavirinae Species 0.000 claims description 6
- 241000315672 SARS coronavirus Species 0.000 claims description 5
- 201000010099 disease Diseases 0.000 claims description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 5
- 241000711443 Bovine coronavirus Species 0.000 claims description 2
- 241000711506 Canine coronavirus Species 0.000 claims description 2
- 241000711475 Feline infectious peritonitis virus Species 0.000 claims description 2
- 241000711467 Human coronavirus 229E Species 0.000 claims description 2
- 241001109669 Human coronavirus HKU1 Species 0.000 claims description 2
- 241000482741 Human coronavirus NL63 Species 0.000 claims description 2
- 241001428935 Human coronavirus OC43 Species 0.000 claims description 2
- 241000711450 Infectious bronchitis virus Species 0.000 claims description 2
- 241000127282 Middle East respiratory syndrome-related coronavirus Species 0.000 claims description 2
- 241001361508 Porcine deltacoronavirus Species 0.000 claims description 2
- 241001135549 Porcine epidemic diarrhea virus Species 0.000 claims description 2
- 241000156302 Porcine hemagglutinating encephalomyelitis virus Species 0.000 claims description 2
- 241000711493 Porcine respiratory coronavirus Species 0.000 claims description 2
- 241000711484 Transmissible gastroenteritis virus Species 0.000 claims description 2
- 208000015181 infectious disease Diseases 0.000 claims description 2
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 39
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 36
- 239000003795 chemical substances by application Substances 0.000 description 35
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 27
- 239000000203 mixture Substances 0.000 description 25
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 21
- LUKZNWIVRBCLON-GXOBDPJESA-N Ciclesonide Chemical compound C1([C@H]2O[C@@]3([C@H](O2)C[C@@H]2[C@@]3(C[C@H](O)[C@@H]3[C@@]4(C)C=CC(=O)C=C4CC[C@H]32)C)C(=O)COC(=O)C(C)C)CCCCC1 LUKZNWIVRBCLON-GXOBDPJESA-N 0.000 description 16
- 239000000243 solution Substances 0.000 description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 14
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 14
- 230000002829 reductive effect Effects 0.000 description 13
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 12
- -1 fatty acid ester Chemical class 0.000 description 12
- 238000012360 testing method Methods 0.000 description 11
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 10
- 238000005160 1H NMR spectroscopy Methods 0.000 description 10
- 230000015572 biosynthetic process Effects 0.000 description 10
- 239000011541 reaction mixture Substances 0.000 description 10
- 238000003786 synthesis reaction Methods 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 9
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 9
- 239000002904 solvent Substances 0.000 description 9
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 238000010898 silica gel chromatography Methods 0.000 description 8
- 239000007787 solid Substances 0.000 description 8
- 229930182555 Penicillin Natural products 0.000 description 7
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 7
- 108020000999 Viral RNA Proteins 0.000 description 7
- 231100000135 cytotoxicity Toxicity 0.000 description 7
- 230000003013 cytotoxicity Effects 0.000 description 7
- 239000012091 fetal bovine serum Substances 0.000 description 7
- 230000002401 inhibitory effect Effects 0.000 description 7
- 229940049954 penicillin Drugs 0.000 description 7
- 229960005322 streptomycin Drugs 0.000 description 7
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 6
- 101800000578 Uridylate-specific endoribonuclease Proteins 0.000 description 6
- 101800001927 Uridylate-specific endoribonuclease nsp15 Proteins 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- 230000003612 virological effect Effects 0.000 description 6
- 101000638154 Homo sapiens Transmembrane protease serine 2 Proteins 0.000 description 5
- 101800001255 Putative 2'-O-methyl transferase Proteins 0.000 description 5
- 102100031989 Transmembrane protease serine 2 Human genes 0.000 description 5
- 241000700605 Viruses Species 0.000 description 5
- 229960003728 ciclesonide Drugs 0.000 description 5
- 239000002537 cosmetic Substances 0.000 description 5
- 239000006210 lotion Substances 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 239000004094 surface-active agent Substances 0.000 description 5
- 230000005727 virus proliferation Effects 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000003446 ligand Substances 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 238000003032 molecular docking Methods 0.000 description 4
- 239000000546 pharmaceutical excipient Substances 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 230000010076 replication Effects 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- RIOQSEWOXXDEQQ-UHFFFAOYSA-N triphenylphosphine Chemical compound C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 RIOQSEWOXXDEQQ-UHFFFAOYSA-N 0.000 description 4
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 3
- 102100031780 Endonuclease Human genes 0.000 description 3
- 108010042407 Endonucleases Proteins 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 229920000881 Modified starch Polymers 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 239000001768 carboxy methyl cellulose Substances 0.000 description 3
- 229920002678 cellulose Polymers 0.000 description 3
- 239000001913 cellulose Substances 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- 239000006071 cream Substances 0.000 description 3
- 230000000249 desinfective effect Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 235000019426 modified starch Nutrition 0.000 description 3
- 235000014593 oils and fats Nutrition 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 238000004088 simulation Methods 0.000 description 3
- FVAUCKIRQBBSSJ-UHFFFAOYSA-M sodium iodide Chemical compound [Na+].[I-] FVAUCKIRQBBSSJ-UHFFFAOYSA-M 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 3
- 238000011282 treatment Methods 0.000 description 3
- WRMNZCZEMHIOCP-UHFFFAOYSA-N 2-phenylethanol Chemical compound OCCC1=CC=CC=C1 WRMNZCZEMHIOCP-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 2
- 229920002785 Croscarmellose sodium Polymers 0.000 description 2
- OKKJLVBELUTLKV-MZCSYVLQSA-N Deuterated methanol Chemical compound [2H]OC([2H])([2H])[2H] OKKJLVBELUTLKV-MZCSYVLQSA-N 0.000 description 2
- ROSDSFDQCJNGOL-UHFFFAOYSA-N Dimethylamine Chemical compound CNC ROSDSFDQCJNGOL-UHFFFAOYSA-N 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 239000012359 Methanesulfonyl chloride Substances 0.000 description 2
- 239000004909 Moisturizer Substances 0.000 description 2
- 101710163270 Nuclease Proteins 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- WNLRTRBMVRJNCN-UHFFFAOYSA-N adipic acid Chemical compound OC(=O)CCCCC(O)=O WNLRTRBMVRJNCN-UHFFFAOYSA-N 0.000 description 2
- 239000003899 bactericide agent Substances 0.000 description 2
- CJZGTCYPCWQAJB-UHFFFAOYSA-L calcium stearate Chemical class [Ca+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O CJZGTCYPCWQAJB-UHFFFAOYSA-L 0.000 description 2
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Chemical compound [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 description 2
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 2
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 2
- OSASVXMJTNOKOY-UHFFFAOYSA-N chlorobutanol Chemical compound CC(C)(O)C(Cl)(Cl)Cl OSASVXMJTNOKOY-UHFFFAOYSA-N 0.000 description 2
- 238000010411 cooking Methods 0.000 description 2
- 235000010947 crosslinked sodium carboxy methyl cellulose Nutrition 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 239000012228 culture supernatant Substances 0.000 description 2
- 231100000263 cytotoxicity test Toxicity 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000003205 fragrance Substances 0.000 description 2
- 125000000524 functional group Chemical group 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- AMXOYNBUYSYVKV-UHFFFAOYSA-M lithium bromide Chemical compound [Li+].[Br-] AMXOYNBUYSYVKV-UHFFFAOYSA-M 0.000 description 2
- KWGKDLIKAYFUFQ-UHFFFAOYSA-M lithium chloride Chemical compound [Li+].[Cl-] KWGKDLIKAYFUFQ-UHFFFAOYSA-M 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- QARBMVPHQWIHKH-UHFFFAOYSA-N methanesulfonyl chloride Chemical compound CS(Cl)(=O)=O QARBMVPHQWIHKH-UHFFFAOYSA-N 0.000 description 2
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 2
- 230000001333 moisturizer Effects 0.000 description 2
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 2
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- RWWYLEGWBNMMLJ-YSOARWBDSA-N remdesivir Chemical compound NC1=NC=NN2C1=CC=C2[C@]1([C@@H]([C@@H]([C@H](O1)CO[P@](=O)(OC1=CC=CC=C1)N[C@H](C(=O)OCC(CC)CC)C)O)O)C#N RWWYLEGWBNMMLJ-YSOARWBDSA-N 0.000 description 2
- RWWYLEGWBNMMLJ-MEUHYHILSA-N remdesivir Drugs C([C@@H]1[C@H]([C@@H](O)[C@@](C#N)(O1)C=1N2N=CN=C(N)C2=CC=1)O)OP(=O)(N[C@@H](C)C(=O)OCC(CC)CC)OC1=CC=CC=C1 RWWYLEGWBNMMLJ-MEUHYHILSA-N 0.000 description 2
- 229920006395 saturated elastomer Polymers 0.000 description 2
- 235000012239 silicon dioxide Nutrition 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 150000003467 sulfuric acid derivatives Chemical class 0.000 description 2
- AYEKOFBPNLCAJY-UHFFFAOYSA-O thiamine pyrophosphate Chemical compound CC1=C(CCOP(O)(=O)OP(O)(O)=O)SC=[N+]1CC1=CN=C(C)N=C1N AYEKOFBPNLCAJY-UHFFFAOYSA-O 0.000 description 2
- 230000009385 viral infection Effects 0.000 description 2
- VBICKXHEKHSIBG-UHFFFAOYSA-N 1-monostearoylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(O)CO VBICKXHEKHSIBG-UHFFFAOYSA-N 0.000 description 1
- QTWJRLJHJPIABL-UHFFFAOYSA-N 2-methylphenol;3-methylphenol;4-methylphenol Chemical compound CC1=CC=C(O)C=C1.CC1=CC=CC(O)=C1.CC1=CC=CC=C1O QTWJRLJHJPIABL-UHFFFAOYSA-N 0.000 description 1
- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 description 1
- 244000215068 Acacia senegal Species 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 241000004176 Alphacoronavirus Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000008904 Betacoronavirus Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 208000001528 Coronaviridae Infections Diseases 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 101710091045 Envelope protein Proteins 0.000 description 1
- 241000008920 Gammacoronavirus Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- DCXXMTOCNZCJGO-UHFFFAOYSA-N Glycerol trioctadecanoate Natural products CCCCCCCCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCCCCCC DCXXMTOCNZCJGO-UHFFFAOYSA-N 0.000 description 1
- 108060003393 Granulin Proteins 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 102100034349 Integrase Human genes 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- 231100000416 LDH assay Toxicity 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 239000000232 Lipid Bilayer Substances 0.000 description 1
- 229920000161 Locust bean gum Polymers 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 108060004795 Methyltransferase Proteins 0.000 description 1
- 241001292005 Nidovirales Species 0.000 description 1
- 102000011931 Nucleoproteins Human genes 0.000 description 1
- 108010061100 Nucleoproteins Proteins 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 229940096437 Protein S Drugs 0.000 description 1
- 101710188315 Protein X Proteins 0.000 description 1
- 239000004373 Pullulan Substances 0.000 description 1
- 229920001218 Pullulan Polymers 0.000 description 1
- 238000013381 RNA quantification Methods 0.000 description 1
- 239000013614 RNA sample Substances 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
- 101710198474 Spike protein Proteins 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 241000008923 Torovirinae Species 0.000 description 1
- 108010067390 Viral Proteins Proteins 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- YKTSYUJCYHOUJP-UHFFFAOYSA-N [O--].[Al+3].[Al+3].[O-][Si]([O-])([O-])[O-] Chemical compound [O--].[Al+3].[Al+3].[O-][Si]([O-])([O-])[O-] YKTSYUJCYHOUJP-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 239000001361 adipic acid Substances 0.000 description 1
- 235000011037 adipic acid Nutrition 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 150000001413 amino acids Chemical group 0.000 description 1
- 239000002260 anti-inflammatory agent Substances 0.000 description 1
- 229940121363 anti-inflammatory agent Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 235000013871 bee wax Nutrition 0.000 description 1
- 239000012166 beeswax Substances 0.000 description 1
- 229960000686 benzalkonium chloride Drugs 0.000 description 1
- CADWTSSKOVRVJC-UHFFFAOYSA-N benzyl(dimethyl)azanium;chloride Chemical compound [Cl-].C[NH+](C)CC1=CC=CC=C1 CADWTSSKOVRVJC-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 235000001465 calcium Nutrition 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 235000013539 calcium stearate Nutrition 0.000 description 1
- 239000008116 calcium stearate Substances 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 150000004649 carbonic acid derivatives Chemical class 0.000 description 1
- 150000001735 carboxylic acids Chemical class 0.000 description 1
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 229960004926 chlorobutanol Drugs 0.000 description 1
- 239000008119 colloidal silica Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 229930003836 cresol Natural products 0.000 description 1
- 229960001681 croscarmellose sodium Drugs 0.000 description 1
- 239000001767 crosslinked sodium carboxy methyl cellulose Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- JAUGGEIKQIHSMF-UHFFFAOYSA-N dialuminum;dimagnesium;dioxido(oxo)silane;oxygen(2-);hydrate Chemical compound O.[O-2].[O-2].[Mg+2].[Mg+2].[Al+3].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O.[O-][Si]([O-])=O JAUGGEIKQIHSMF-UHFFFAOYSA-N 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- MOTZDAYCYVMXPC-UHFFFAOYSA-N dodecyl hydrogen sulfate Chemical class CCCCCCCCCCCCOS(O)(=O)=O MOTZDAYCYVMXPC-UHFFFAOYSA-N 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- BEFDCLMNVWHSGT-UHFFFAOYSA-N ethenylcyclopentane Chemical compound C=CC1CCCC1 BEFDCLMNVWHSGT-UHFFFAOYSA-N 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- ZCGNOVWYSGBHAU-UHFFFAOYSA-N favipiravir Chemical compound NC(=O)C1=NC(F)=CNC1=O ZCGNOVWYSGBHAU-UHFFFAOYSA-N 0.000 description 1
- 229950008454 favipiravir Drugs 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 1
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 1
- UFVKGYZPFZQRLF-UHFFFAOYSA-N hydroxypropyl methyl cellulose Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 UFVKGYZPFZQRLF-UHFFFAOYSA-N 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- LSACYLWPPQLVSM-UHFFFAOYSA-N isobutyric acid anhydride Chemical compound CC(C)C(=O)OC(=O)C(C)C LSACYLWPPQLVSM-UHFFFAOYSA-N 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 238000002843 lactate dehydrogenase assay Methods 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 235000010420 locust bean gum Nutrition 0.000 description 1
- 239000000711 locust bean gum Substances 0.000 description 1
- 229940031703 low substituted hydroxypropyl cellulose Drugs 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 229960003511 macrogol Drugs 0.000 description 1
- 229940037627 magnesium lauryl sulfate Drugs 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- HBNDBUATLJAUQM-UHFFFAOYSA-L magnesium;dodecyl sulfate Chemical compound [Mg+2].CCCCCCCCCCCCOS([O-])(=O)=O.CCCCCCCCCCCCOS([O-])(=O)=O HBNDBUATLJAUQM-UHFFFAOYSA-L 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 239000004292 methyl p-hydroxybenzoate Substances 0.000 description 1
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 229960002216 methylparaben Drugs 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 238000001821 nucleic acid purification Methods 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- WVDDGKGOMKODPV-ZQBYOMGUSA-N phenyl(114C)methanol Chemical compound O[14CH2]C1=CC=CC=C1 WVDDGKGOMKODPV-ZQBYOMGUSA-N 0.000 description 1
- 229940067107 phenylethyl alcohol Drugs 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 229920001495 poly(sodium acrylate) polymer Polymers 0.000 description 1
- 239000000244 polyoxyethylene sorbitan monooleate Substances 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 229940068968 polysorbate 80 Drugs 0.000 description 1
- 229920001592 potato starch Polymers 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 239000004405 propyl p-hydroxybenzoate Substances 0.000 description 1
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 description 1
- 229960003415 propylparaben Drugs 0.000 description 1
- 235000019423 pullulan Nutrition 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 150000004760 silicates Chemical class 0.000 description 1
- RMAQACBXLXPBSY-UHFFFAOYSA-N silicic acid Chemical compound O[Si](O)(O)O RMAQACBXLXPBSY-UHFFFAOYSA-N 0.000 description 1
- 125000005624 silicic acid group Chemical class 0.000 description 1
- NGHMEZWZOZEZOH-UHFFFAOYSA-N silicic acid;hydrate Chemical compound O.O[Si](O)(O)O NGHMEZWZOZEZOH-UHFFFAOYSA-N 0.000 description 1
- 239000000344 soap Substances 0.000 description 1
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical compound [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 description 1
- 239000004299 sodium benzoate Substances 0.000 description 1
- 235000010234 sodium benzoate Nutrition 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 235000009518 sodium iodide Nutrition 0.000 description 1
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 1
- NNMHYFLPFNGQFZ-UHFFFAOYSA-M sodium polyacrylate Chemical compound [Na+].[O-]C(=O)C=C NNMHYFLPFNGQFZ-UHFFFAOYSA-M 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 239000013042 solid detergent Substances 0.000 description 1
- 239000004334 sorbic acid Substances 0.000 description 1
- 235000010199 sorbic acid Nutrition 0.000 description 1
- 229940075582 sorbic acid Drugs 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000013595 supernatant sample Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 description 1
- 229940033663 thimerosal Drugs 0.000 description 1
- 231100000041 toxicology testing Toxicity 0.000 description 1
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 230000029812 viral genome replication Effects 0.000 description 1
- 239000001993 wax Substances 0.000 description 1
- 239000000230 xanthan gum Substances 0.000 description 1
- 235000010493 xanthan gum Nutrition 0.000 description 1
- 229920001285 xanthan gum Polymers 0.000 description 1
- 229940082509 xanthan gum Drugs 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Steroid Compounds (AREA)
Description
本発明は、新規化合物、及び、抗コロナウイルス剤に関し、特にSARS-CoV-2に対する抗ウイルス作用を有する抗コロナウイルス剤に関する。 The present invention relates to a novel compound and an anti-coronavirus agent, in particular an anti-coronavirus agent having an antiviral effect against SARS-CoV-2.
SARS-CoV-2は、2019年12月に中国湖北省武漢市で発生した肺炎患者から検出された新型コロナウイルスであり(非特許文献１)、その後全世界に拡散し、2020年3月には世界保健機関よりパンデミック宣言がなされた。2020年10月現在，感染者は3740万人，死者は108万人を超えている。新型コロナウイルス感染症(COVID-19)は人類にとって蔓延を抑制すべき疾患である。 SARS-CoV-2 is a novel coronavirus that was detected in a pneumonia patient in Wuhan, Hubei Province, China in December 2019 (Non-Patent Document 1), and has since spread throughout the world, with the World Health Organization declaring it a pandemic in March 2020. As of October 2020, the number of infected people is 37.4 million, and the death toll exceeds 1.08 million. Novel coronavirus disease (COVID-19) is a disease that humanity must contain in its spread.
SARS-CoV-2は一本鎖プラス鎖RNAウイルスであり、ウイルス粒子表面のエンベロープ(脂質二重膜)に、花弁状のスパイク蛋白(S)の突起が存在する。SARS-CoV-2は更にエンベロープ蛋白(E)、マトリックス蛋白(M)、核蛋白(N)によって構成されている。 SARS-CoV-2 is a single-stranded positive-stranded RNA virus with petal-shaped spike protein (S) protrusions on the envelope (lipid bilayer) on the surface of the virus particle. SARS-CoV-2 is further composed of envelope protein (E), matrix protein (M), and nucleoprotein (N).
世界中の研究者により治療薬やワクチンの開発が行われており、いくつかの市販薬（レムデシビル，シクレソニドCicle，ファビピラビル等）がCOVID-19の治療に有効であることが示唆されている。一般的な抗ウイルス薬は、ウイルスのポリメラーゼあるいはヌクレアーゼを阻害することで薬効し、2020年5月にCOVID-19治療薬として国内承認されたレムデシビルは、ポリメラーゼを阻害することでウイルスの複製を抑制する。一方で現在のところ、COVID-19のヌクレアーゼ阻害薬は承認されていないが、最近、シクレソニド活性代謝物Cic2がSARS-CoV-2のエンドヌクレアーゼ（Nsp15）に結合してゲノムRNAの複製を抑制することが報告された(非特許文献２，３)。Nsp15は宿主の免疫システムからウイルス自身を守る酵素として働くこと、また複数のコロナウイルスにおいて構造的類似性があること（2003年に流行したSARS-CoVのNsp15とは95.7%の相同性）が知られている(非特許文献４)。 Researchers around the world are developing treatments and vaccines, and it has been suggested that some commercially available drugs (remdesivir, ciclesonide, favipiravir, etc.) are effective in treating COVID-19. General antiviral drugs work by inhibiting viral polymerase or nuclease, and remdesivir, which was approved in Japan in May 2020 as a treatment for COVID-19, inhibits viral replication by inhibiting polymerase. On the other hand, no nuclease inhibitors have been approved for COVID-19 at present, but it has recently been reported that the active metabolite of ciclesonide, Cic2, binds to the endonuclease (Nsp15) of SARS-CoV-2 and inhibits the replication of genomic RNA (Non-Patent Documents 2 and 3). It is known that Nsp15 acts as an enzyme that protects the virus itself from the host's immune system, and that it has structural similarities in multiple coronaviruses (95.7% homology with Nsp15 of SARS-CoV, which was prevalent in 2003) (Non-Patent Document 4).
しかしながらシクレソニドCicle及びシクレソニド活性代謝物Cic2の抗ウイルス作用はいずれも十分ではない。 However, the antiviral effects of both ciclesonide (Cicle) and its active metabolite (Cic2) are insufficient.
本発明はかかる問題点に鑑みてなされたものであって、十分な抗ウイルス作用を有する抗コロナウイルス剤を提供することを目的とする。 The present invention was made in consideration of these problems, and aims to provide an anti-coronavirus agent with sufficient antiviral activity.
本発明にかかる新規化合物は下記構造式からなる。 The novel compound of the present invention has the following structural formula:
本発明にかかる抗コロナウイルス剤は、下記化合物を有効成分とし、コロナウイルスに対する抗ウイルス作用を有することを特徴とする。 The anti-coronavirus agent of the present invention contains the following compound as an active ingredient and is characterized by having an antiviral effect against coronaviruses.
本発明にかかる抗コロナウイルス剤は、下記化合物を有効成分とし、コロナウイルスに対する抗ウイルス作用を有することを特徴とする。 The anti-coronavirus agent of the present invention contains the following compound as an active ingredient and is characterized by having an antiviral effect against coronaviruses.
本発明にかかる抗コロナウイルス剤は、下記化合物を有効成分とし、コロナウイルスに対する抗ウイルス作用を有することを特徴とする。 The anti-coronavirus agent of the present invention contains the following compound as an active ingredient and is characterized by having an antiviral effect against coronaviruses.
本発明にかかる抗コロナウイルス剤は、下記化合物を有効成分とし、コロナウイルスに対する抗ウイルス作用を有することを特徴とする。 The anti-coronavirus agent of the present invention contains the following compound as an active ingredient and is characterized by having an antiviral effect against coronaviruses.
本発明によれば、十分な抗ウイルス作用を有する抗コロナウイルス剤が得られる。 The present invention provides an anti-coronavirus agent with sufficient antiviral activity.
以下、添付の図面を参照して本発明の実施形態について具体的に説明するが、当該実施形態は本発明の原理の理解を容易にするためのものであり、本発明の範囲は、下記の実施形態に限られるものではなく、当業者が以下の実施形態の構成を適宜置換した他の実施形態も、本発明の範囲に含まれる。 The following describes in detail an embodiment of the present invention with reference to the attached drawings. However, this embodiment is intended to facilitate understanding of the principles of the present invention, and the scope of the present invention is not limited to the following embodiment. Other embodiments in which a person skilled in the art appropriately replaces the configuration of the following embodiment are also included in the scope of the present invention.
(1)新規化合物
本発明にかかる新規化合物は、下記構造式からなる。
(1) Novel Compound The novel compound according to the present invention has the following structural formula:
上記シクレソニド類縁体Cic4は下記ルートにて合成することが可能である。 The above ciclesonide analog Cic4 can be synthesized via the following route.
Cic2，トリエチルアミンのジクロロメタン溶液に、塩化メタンスルホニルのジクロロメタン溶液を加える。室温にて撹拌した後、反応液をジクロロメタンで希釈し、飽和炭酸水素ナトリウム水溶液、飽和食塩水で洗浄する。無水硫酸ナトリウム上で乾燥後、ろ過して溶媒を減圧留去した。得られた残渣を精製し、Cic3を得る。 Cic2, a solution of triethylamine in dichloromethane is mixed with a solution of methanesulfonyl chloride in dichloromethane. After stirring at room temperature, the reaction mixture is diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate and saturated saline. After drying over anhydrous sodium sulfate, it is filtered and the solvent is removed under reduced pressure. The resulting residue is purified to obtain Cic3.
次にCic3のアセトニトリル溶液にアジ化ナトリウムを加え、攪拌した。反応液を酢酸エチルで希釈し、水、飽和食塩水で洗浄、無水硫酸ナトリウム上で乾燥後、ろ過して溶媒を減圧留去する。得られた残渣を精製し、Cic4を得る。 Next, sodium azide was added to the acetonitrile solution of Cic3 and stirred. The reaction solution was diluted with ethyl acetate, washed with water and saturated saline, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The resulting residue was purified to obtain Cic4.
(2)抗コロナウイルス剤
SARS-CoVには非構造タンパク質15（nsp15）として知られる特定の遺伝子配列があり、その配列はSARS-CoV-2の遺伝子配列と95.7％の類似性があることが分かっている。nsp15の配列及び構造を検証したところ、nsp15は病気の発症に欠かせないエンドヌクレアーゼ酵素で、複数のコロナウイルスにおいて構造的類似性があることが判明している。このエンドヌクレアーゼは、高い確率で宿主の免疫システムからコロナウイルス自身を守るという共通の機能を果たす。nsp15阻害薬はSARS-CoVやSARS-CoV-2だけではなく、あらゆる新種のコロナウイルスに対して有効であると考えられる。
(2) Anti-coronavirus agents
SARS-CoV has a specific gene sequence known as nonstructural protein 15 (nsp15), which has been shown to have 95.7% similarity to the gene sequence of SARS-CoV-2. Examination of the sequence and structure of nsp15 has revealed that nsp15 is an endonuclease enzyme essential for disease development and has structural similarities in multiple coronaviruses. This endonuclease has a high probability of performing a common function in protecting coronaviruses from the host immune system. It is believed that nsp15 inhibitors will be effective not only against SARS-CoV and SARS-CoV-2, but also against any new coronaviruses.
新型コロナウイルスSARS-CoV-2のゲノム複製に関与するウイルスタンパク質とシクレソニドとの相互作用を先端バイオインフォマティクス技術ならびに高性能コンピューターを駆使したドッキングシミュレーションを用い、シクレソニド誘導体Cic2の結合解析を検討した。リガンドの結合解析からタンパク質の結合に重要であると考えられるSer294及びPro344に対応する一級水酸基を各官能基に置換したシクレソニド類縁体Cic3～Cic11を設計した(図１)。なおシクレソニドは、新型コロナウイルスゲノム複製酵素（RNA依存性RNAポリメラーゼ）には作用しなかったが、nsp15の活性中心と結合し、ウイルスゲノムの正確な複製を阻害することが判明している。 The binding analysis of the ciclesonide derivative Cic2 was carried out using docking simulations that make full use of advanced bioinformatics technology and high-performance computers to investigate the interaction between ciclesonide and viral proteins involved in the genome replication of the novel coronavirus SARS-CoV-2. Based on the ligand binding analysis, we designed ciclesonide analogs Cic3 to Cic11 in which the primary hydroxyl groups corresponding to Ser294 and Pro344, which are thought to be important for protein binding, were replaced with the respective functional groups (Figure 1). Although ciclesonide did not act on the novel coronavirus genome replication enzyme (RNA-dependent RNA polymerase), it was found to bind to the active center of nsp15 and inhibit the accurate replication of the viral genome.
本発明にかかる抗コロナウイルス剤は、下記化合物を有効成分とし、コロナウイルスに対する抗ウイルス作用を有する。 The anti-coronavirus agent of the present invention contains the following compound as an active ingredient and has an antiviral effect against coronaviruses.
本発明にかかる抗コロナウイルス剤は、下記化合物を有効成分とし、コロナウイルスに対する抗ウイルス作用を有する。 The anti-coronavirus agent of the present invention contains the following compound as an active ingredient and has an antiviral effect against coronaviruses.
本発明にかかる抗コロナウイルス剤は、下記化合物を有効成分とし、コロナウイルスに対する抗ウイルス作用を有する。 The anti-coronavirus agent of the present invention contains the following compound as an active ingredient and has an antiviral effect against coronaviruses.
本発明にかかる抗コロナウイルス剤は、下記化合物を有効成分とし、コロナウイルスに対する抗ウイルス作用を有する。 The anti-coronavirus agent of the present invention contains the following compound as an active ingredient and has an antiviral effect against coronaviruses.
本発明にかかる抗コロナウイルス剤が対象とするコロナウイルスは、特に限定されるわけではないが、例えば野生型コロナウイルスの変種である。 The coronaviruses targeted by the anti-coronavirus agent of the present invention are not particularly limited, but include, for example, variants of wild-type coronaviruses.
本発明にかかる抗コロナウイルス剤が対象とするコロナウイルスは、特に限定されるわけではないが、例えば野生型コロナウイルスである。 The coronavirus targeted by the anti-coronavirus agent of the present invention is not particularly limited, but is, for example, a wild-type coronavirus.
野生型コロナウイルスは、コロナウイルス亜科に属するコロナウイルスからなる。コロナウイルスはニドウイルス目に分類され、その中にコロナウイルス科、そして、コロナウイルス亜科とトロウイルス亜科に分類される。なおコロナウイルス亜科はさらに、アルファコロナウイルス属、ベータコロナウイルス属、ガンマコロナウイルス属の３つの属に分類される。 Wild-type coronaviruses are made up of coronaviruses that belong to the Coronavirinae subfamily. Coronaviruses are classified in the order Nidovirales, which are divided into the Coronaviridae family and the subfamily Coronavirinae and Torovirinae. The Coronavirinae subfamily is further divided into three genera: Alphacoronavirus, Betacoronavirus, and Gammacoronavirus.
コロナウイルス亜科に属するコロナウイルスは、特に限定されるわけではないが、例えばSARS-CoV-2である。 An example of a coronavirus belonging to the Coronavirinae subfamily is, but is not limited to, SARS-CoV-2.
コロナウイルス亜科に属するコロナウイルスは、特に限定されるわけではないが、例えばSARSコロナウイルス、MERSコロナウイルス、ヒトコロナウイルス229E、ヒトコロナウイルスOC43、ヒトコロナウイルスHKU1、ヒトコロナウイルスNL63、ネコ伝染性腹膜炎ウイルス、イヌコロナウイルス、鶏伝染性気管支炎ウイルス、ウシコロナウイルス、及び、伝染性胃腸炎ウイルス、ブタデルタコロナウイルス、ブタ流行性下痢ウイルス、ブタ呼吸器型コロナウイルス、ブタ血球凝集性脳脊髄炎コロナウイルスを含むブタコロナウイルスからなる群から選択される。 The coronavirus belonging to the Coronavirinae subfamily is, but is not limited to, selected from the group consisting of SARS coronavirus, MERS coronavirus, human coronavirus 229E, human coronavirus OC43, human coronavirus HKU1, human coronavirus NL63, feline infectious peritonitis virus, canine coronavirus, avian infectious bronchitis virus, bovine coronavirus, and porcine coronaviruses including transmissible gastroenteritis virus, porcine deltacoronavirus, porcine epidemic diarrhea virus, porcine respiratory coronavirus, and porcine hemagglutinating encephalomyelitis coronavirus.
本発明にかかる抗コロナウイルス剤は、抗コロナウイルス活性を有しており、抗コロナウイルス活性とは、コロナウイルスを死滅させる場合、又は、コロナウイルスは残存していてもその表面タンパク質に作用して感染能力（増殖能力）を喪失させる場合の少なくとも何れか一方を含むものとする。 The anti-coronavirus agent of the present invention has anti-coronavirus activity, which includes at least one of killing the coronavirus or acting on the surface protein of the coronavirus to cause it to lose its infectivity (ability to grow) even if the coronavirus remains.
本発明にかかる抗コロナウイルス剤と、アルコール、界面活性剤、抗菌剤、保湿剤及び化粧品用油脂類からなる群より選ばれた少なくとも１つの成分と、を含有することで抗コロナウイルス用組成物と規定することも可能である。 It is also possible to define it as an anti-coronavirus composition by containing the anti-coronavirus agent of the present invention and at least one component selected from the group consisting of alcohol, surfactant, antibacterial agent, moisturizer, and cosmetic oils and fats.
その態様は特に限定されるものではないが、代表的には以下のようなものが挙げられる。
・抗コロナウイルス剤とアルコールとを含有するアルコール製剤
・抗コロナウイルス剤と界面活性剤とを含有する洗浄用組成物
・抗コロナウイルス剤と抗菌剤とを含有する消毒用組成物
・抗コロナウイルス剤と保湿剤及び／又は化粧品用油脂類とを含有するローション、乳液又はクリーム
洗浄用組成物は、食品、食器、調理器具、作業者の手指や着衣等の汚れを落とすとともにコロナウイルスを消毒することができる態様の組成物であり、例えば液状又は固形状の洗剤として提供される。アルコール製剤及び消毒用組成物は、食品、食器、調理器具、作業者の手指、あるいはコロナウイルス患者の汚物を取り扱った器具等に付着した、コロナウイルス及び細菌類を不活性化するために使用される態様の組成物であり、例えば従来のエタノール製剤と同様の噴霧剤として提供される。ローション、クリーム及び乳液は、水仕事等で荒れやすい作業者の手指に塗ってスキンケアをするとともにコロナウイルスを消毒することができる態様の組成物（基礎化粧品）である。
The form is not particularly limited, but typical examples include the following:
・Alcohol preparations containing an anti-coronavirus agent and alcohol・Cleaning compositions containing an anti-coronavirus agent and a surfactant・Disinfecting compositions containing an anti-coronavirus agent and an antibacterial agent・Lotions, milky lotions, or creams containing an anti-coronavirus agent and a moisturizing agent and/or cosmetic oils and fats The cleaning compositions are compositions capable of removing dirt from food, tableware, cooking utensils, the hands and fingers of workers, clothing, etc., and disinfecting coronaviruses, and are provided, for example, as liquid or solid detergents. The alcohol preparations and disinfecting compositions are compositions used to inactivate coronaviruses and bacteria attached to food, tableware, cooking utensils, the hands and fingers of workers, or utensils that have handled coronavirus patient dirt, and are provided, for example, as sprays similar to conventional ethanol preparations. The lotions, creams, and milky lotions are compositions (basic cosmetics) that can be applied to the hands and fingers of workers that are easily roughened by water work, etc., to provide skin care and disinfect coronaviruses.
なお、上述の抗コロナウイルス用組成物において、アルコール、界面活性剤、殺菌剤、保湿剤、及び化粧品用油脂類は２種以上を組み合わせて用いてもよい。例えばアルコール製剤については、抗菌性をより高めるために脂肪酸エステル等の界面活性剤を更に配合することも好ましい。また、上記洗浄用組成物は、界面活性剤に加えて殺菌剤やアルコールを配合したハンドソープ等の態様をとることができ、上記クリームは、手肌を保護する成分と共に手肌を清浄に保つための抗菌剤やアルコールを配合した態様をとることができる。 In the above-mentioned anti-coronavirus composition, the alcohol, surfactant, bactericide, moisturizer, and cosmetic oils and fats may be used in combination of two or more kinds. For example, it is preferable to further blend a surfactant such as a fatty acid ester in an alcohol preparation to further enhance antibacterial properties. The above-mentioned cleaning composition can take the form of a hand soap containing a bactericide and alcohol in addition to a surfactant, and the above-mentioned cream can take the form of a compound containing an antibacterial agent and alcohol to keep the hands clean together with a component to protect the hands.
上述の抗コロナウイルス用組成物には、所望の性能を賦与し各組成物の品質を高めるための各種成分、例えば増粘剤（キサンタンガム、ローカストビーンガム、ポリアクリル酸ナトリウム等）、酸化防止剤、香料、色素等、またローション等の化粧品にあっては肌荒れ防止剤、消炎剤等を、適宜配合することができる。 The above-mentioned anti-coronavirus compositions can be appropriately blended with various ingredients to impart desired properties and improve the quality of each composition, such as thickeners (xanthan gum, locust bean gum, sodium polyacrylate, etc.), antioxidants, fragrances, colorants, etc., and in the case of cosmetics such as lotions, skin care agents, anti-inflammatory agents, etc.
上述の抗コロナウイルス用組成物における抗コロナウイルス剤の含有量は、抗コロナウイルス活性が発現される範囲において、組成物の成分構成や使用方法等の態様に応じて適宜調整することができる。 The content of the anti-coronavirus agent in the above-mentioned anti-coronavirus composition can be appropriately adjusted according to the composition's component composition, method of use, and other aspects, within the range in which anti-coronavirus activity is expressed.
本発明の抗コロナウイルス剤の有効成分濃度は、抗ノロウイルス活性が発現される範囲において適宜調整することができ、例えば１μM～80μMである。 The concentration of the active ingredient of the anti-coronavirus agent of the present invention can be appropriately adjusted within the range in which anti-norovirus activity is expressed, for example, 1 μM to 80 μM.
化合物Cic4を有効成分とする抗コロナウイルス剤では、例えば5μM～80μMであり、好ましくは7μM～40μMであり、より好ましくは6μM～20μMである。 For anti-coronavirus agents containing the compound Cic4 as an active ingredient, the concentration is, for example, 5 μM to 80 μM, preferably 7 μM to 40 μM, and more preferably 6 μM to 20 μM.
化合物Cic6を有効成分とする抗コロナウイルス剤では、例えば1μM～80μMであり、好ましくは5μM～40μMであり、より好ましくは6μM～20μMである。 For anti-coronavirus agents containing the compound Cic6 as an active ingredient, the concentration is, for example, 1 μM to 80 μM, preferably 5 μM to 40 μM, and more preferably 6 μM to 20 μM.
本発明の抗コロナウイルス剤は、コロナウイルスの感染を予防及び／又は前記コロナウイルスによる疾患を治療するための医薬用の抗コロナウイルス剤として規定することも可能である。 The anti-coronavirus agent of the present invention can also be defined as a medicinal anti-coronavirus agent for preventing coronavirus infection and/or treating a disease caused by the coronavirus.
医薬用の抗コロナウイルス剤は、投与部位として、経口投与、口腔内投与、気道内投与、皮下投与、筋肉内投与、血管内（静脈内）投与等を挙げることができる。本発明の抗コロナウイルス剤を医薬として使用する場合、公知の方法により種々の態様に製剤化され、例えば、注射剤、カプセル剤、錠剤、シロップ剤、顆粒剤、貼布剤、点滴、軟膏等を挙げることができる。本発明の抗コロナウイルス剤は、単独で投与しても良いし、薬理学的に許容される単体と共に投与されてもよい。 The administration site of the anti-coronavirus agent for pharmaceutical use can include oral administration, oral administration, intratracheal administration, subcutaneous administration, intramuscular administration, and intravascular (intravenous) administration. When the anti-coronavirus agent of the present invention is used as a pharmaceutical, it is formulated into various forms by known methods, such as injections, capsules, tablets, syrups, granules, patches, drips, ointments, and the like. The anti-coronavirus agent of the present invention can be administered alone or together with a pharmacologically acceptable carrier.
これらの製剤は、賦形剤（例えば、乳糖、白糖、ブドウ糖、マンニット、ソルビットのような糖誘導体；トウモロコシデンプン、馬鈴薯デンプン、α－デンプン、デキストリン、カルボキシメチルデンプンのようなデンプン誘導体；結晶セルロース、低置換度ヒドロキシプロピルセルロース、ヒドロキシプロピルメチルセルロース、カルボキシメチルセルロース、カルボキシメチルセルロースカルシウム、内部架橋カルボキシメチルセルロースナトリウムのようなセルロース誘導体；アラビアゴム；デキストラン；プルラン；軽質無水珪酸、合成珪酸アルミニウム、メタ珪酸アルミン酸マグネシウムのような珪酸塩誘導体；リン酸カルシウムのようなリン酸塩誘導体；炭酸カルシウムのような炭酸塩誘導体；硫酸カルシウムのような硫酸塩誘導体等）、結合剤（例えば、前記の賦形剤；ゼラチン；ポリビニルピロリドン；マクロゴール等）、崩壊剤（例えば、前記の賦形剤；クロスカルメロースナトリウム、カルボキシメチルスターチナトリウム、架橋ポリビニルピロリドンのような化学修飾された、デンプン、セルロース誘導体等）、滑沢剤（例えば、タルク；ステアリン酸；ステアリン酸カルシウム、ステアリン酸マグネシウムのようなステアリン酸金属塩；コロイドシリカ；ビーガム；ビ－ズワックス、ゲイロウのようなワックス類；硼酸；グリコール；フマル酸；アジピン酸のようなカルボン酸類：安息香酸ナトリウムのようなカルボン酸ナトリウム塩；硫酸ナトリウムのような硫酸類塩；ロイシン；ラウリル硫酸ナトリウム、ラウリル硫酸マグネシウムのようなラウリル硫酸塩；無水珪酸、珪酸水和物のような珪酸類；前記の賦形剤におけるデンプン誘導体等）、安定剤（例えば、メチルパラペン、プロピルパラペンのようなパラオキシ安息香酸エステル類；クロロブタノール、ベンジルアルコール、フェニルエチルアルコールのようなアルコール類；塩化ベンザルコニウム；フェノール；クレゾールのようなフェノール類；チメロサール；無水酢酸；ソルビン酸等）、矯味矯臭剤（例えば、通常使用される、甘味料、酸味料、香料等）、懸濁化剤（例えば、ポリソルベート80、カルボキシメチルセルロースナトリウム等）、希釈剤、製剤用溶剤（例えば、水、エタノール、グリセリン等）等の添加物を用いて周知の方法で製造される。また、本発明の抗コロナウイルス剤を注射剤として使用する場合、保存容器としては、アンプル、バイアル、プレフィルドシリンジ、ペン型注射器用カートリッジ、及び、点滴用バッグ等を挙げることができる。 These preparations contain excipients (e.g., sugar derivatives such as lactose, sucrose, glucose, mannitol, and sorbitol; starch derivatives such as corn starch, potato starch, α-starch, dextrin, and carboxymethyl starch; cellulose derivatives such as crystalline cellulose, low-substituted hydroxypropylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, calcium carboxymethylcellulose, and internally crosslinked sodium carboxymethylcellulose; gum arabic; dextran; pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, and magnesium aluminometasilicate; phosphate derivatives such as calcium phosphate; carbonate derivatives such as calcium carbonate; sulfate derivatives such as calcium sulfate, etc.), binders (e.g., the above-mentioned excipients; gelatin; polyvinylpyrrolidone; macrogol, etc.), disintegrants (e.g., the above-mentioned excipients; chemically modified starches and cellulose derivatives such as croscarmellose sodium, sodium carboxymethyl starch, and crosslinked polyvinylpyrrolidone, etc.), lubricants (e.g., talc; stearin, Acids; metal stearates such as calcium stearate and magnesium stearate; colloidal silica; veegum; waxes such as beeswax and gazel; boric acid; glycol; fumaric acid; carboxylic acids such as adipic acid; sodium carboxylates such as sodium benzoate; sulfates such as sodium sulfate; leucine; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acids such as silicic anhydride and silicic acid hydrate; starch derivatives in the above-mentioned excipients, etc.), stabilizers (e.g., methylparaben, proline, etc.), The anti-coronavirus agent of the present invention is produced by a known method using additives such as paraoxybenzoic acid esters such as propylparaben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; phenol; phenols such as cresol; thimerosal; acetic anhydride; sorbic acid, etc.), flavoring agents (e.g., commonly used sweeteners, acidulants, fragrances, etc.), suspending agents (e.g., polysorbate 80, sodium carboxymethylcellulose, etc.), diluents, and formulation solvents (e.g., water, ethanol, glycerin, etc.). When the anti-coronavirus agent of the present invention is used as an injection, examples of the storage container include ampoules, vials, prefilled syringes, cartridges for pen-type syringes, and infusion bags.
本発明の抗コロナウイルス剤の医薬としての投与量は、所望の治療効果又は予防効果が得られる投与量であれば特に限定は無く、症状、性別、年齢等により適宜決定することができる。投与量として、好ましくは、1ng/kg～10mg/kgであり、より好ましくは、10ng/kg～1mg/kgであり、更に好ましくは、5～500μg/kgであり、より更に好ましくは、10～100μg/kgであり、最も好ましくは、10～30μg/kgである。 The dosage of the anti-coronavirus agent of the present invention as a medicine is not particularly limited as long as it is a dosage that provides the desired therapeutic or preventive effect, and can be appropriately determined depending on symptoms, sex, age, etc. The dosage is preferably 1 ng/kg to 10 mg/kg, more preferably 10 ng/kg to 1 mg/kg, even more preferably 5 to 500 μg/kg, even more preferably 10 to 100 μg/kg, and most preferably 10 to 30 μg/kg.
(1)シクレソニド類縁体の分子設計
X線結晶構造が明らかとなっているNsp15（PDB：6VWW）に対し、 MOE（Molecular Operating Environment；CCG社）を用いてシクレソニド代謝物Cic2のドッキングスタディを行った。
(1) Molecular design of ciclesonide analogues
A docking study of the ciclesonide metabolite Cic2 was performed using MOE (Molecular Operating Environment; CCG) with Nsp15 (PDB: 6VWW), whose X-ray crystal structure has been revealed.
始めに、結晶水の消去、タンパク質への水素原子の付加、アミノ酸側鎖構造の補正を行い、Site Finderを用いてリガンド結合領域を検出した。次いで、Nsp15とCic2とのドッキングシミュレーション（力場；AMBER10:EHT）により、リガンドの結合様式解析を行った（図1）。リガンドの結合様式からタンパク質の結合に重要であると考えられるSer294及びPro344に対応する一級水酸基を各官能基に置換したシクレソニド類縁体Cic3-Cic11を設計した。 First, the crystal water was removed, hydrogen atoms were added to the protein, and the amino acid side chain structure was corrected, and the ligand-binding region was detected using Site Finder. Next, the ligand-binding mode was analyzed by a docking simulation (force field: AMBER10:EHT) between Nsp15 and Cic2 (Figure 1). Based on the ligand-binding mode, we designed ciclesonide analogs Cic3-Cic11 in which the primary hydroxyl groups corresponding to Ser294 and Pro344, which are thought to be important for protein binding, were replaced with the respective functional groups.
(2)シクレソニド類縁体の合成
次にシクレソニド類縁体Cic3～Cic11を下記に示すスキームにてCic2を出発原料として合成した。
(2) Synthesis of Ciclesonide Analogues Next, ciclesonide analogues Cic3 to Cic11 were synthesized from Cic2 as a starting material according to the scheme shown below.
(Cic3の合成 )
Cic2 (250 mg, 0.5 mmol)，トリエチルアミン (154 μL, 1.1 mmol)のジクロロメタン溶液 (2 mL)に、0℃にて塩化メタンスルホニル (43 μL, 0.55 mmol)のジクロロメタン溶液 (0.5 mL)を加えた。室温にて30分間撹拌した後、反応液をジクロロメタンで希釈し、飽和炭酸水素ナトリウム水溶液、飽和食塩水で洗浄した。無水硫酸ナトリウム上で乾燥後、ろ過して溶媒を減圧留去した。得られた残渣をフラッシュシリカゲルカラムクロマトグラフィー （ヘキサン/酢酸エチル = 4 : 1 to 1 : 1）にて精製し、Cic3を無色泡状固体として得た (270 mg, 99%)。
1H NMR (600 MHz, CDCl3) : δ 7.25 (d, J = 10.2 Hz, 1H), 6.29 (dd, J = 10.2, 1.2 Hz, 1H), 6.03 (s, 1H), 5.00 (d, J = 1.8 Hz, 2H), 4.86 (d, J = 4.8 Hz, 1H), 4.51 (t, J = 3.0 Hz, 1H), 4.31 (d, J = 4.8 Hz, 1H), 3.25 (s, 3H), 2.57 (ddd, J = 13.2, 13.2, 4.8 Hz, 1H), 2.35 (dd, J = 13.2, 3.0 Hz, 1H), 2.20-2.14 (m, 1H), 2.08-2.04 (m, 1H), 1.79-1.70 (m, 3H), 1.66-1.56 (m, 9H), 1.45 (s, 3H), 1.26-1.04 (m, 7H), 0.96 (s, 3H).
13C NMR (151 MHz, CDCl3) : δ 202.8, 186.5, 169.5, 155.8, 128.0, 122.6, 107.7, 97.2, 81.9, 71.6, 69.9, 55.0, 49.7, 46.1, 43.9, 40.9, 40.6, 39.5, 33.9, 33.3, 31.8, 30.3, 27.1, 26.2, 25.5, 21.1, 17.1.
(Synthesis of Cic3)
To a dichloromethane solution (2 mL) of Cic2 (250 mg, 0.5 mmol) and triethylamine (154 μL, 1.1 mmol), a dichloromethane solution (0.5 mL) of methanesulfonyl chloride (43 μL, 0.55 mmol) was added at 0°C. After stirring at room temperature for 30 minutes, the reaction mixture was diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate and saturated saline. After drying over anhydrous sodium sulfate, the mixture was filtered and the solvent was removed under reduced pressure. The resulting residue was purified by flash silica gel column chromatography (hexane/ethyl acetate = 4 : 1 to 1 : 1) to give Cic3 as a colorless foamy solid (270 mg, 99%).
1H NMR (600 MHz, CDCl3): δ 7.25 (d, J = 10.2 Hz, 1H), 6.29 (dd, J = 10.2, 1.2 Hz, 1H), 6.03 (s, 1H), 5.00 (d, J = 1.8 Hz, 2H), 4.86 (d, J = 4.8 Hz, 1H), 4.51 (t, J = 3.0 Hz, 1H), 4.31 (d, J = 4.8 Hz, 1H), 3.25 (s, 3H), 2.57 (ddd, J = 13.2, 13.2, 4.8 Hz, 1H), 2.35 (dd, J = 13.2, 3.0Hz, 1H), 2.20-2.14 (m, 1H), 2.08-2.04 (m, 1H), 1.79-1.70 (m, 3H), 1.66-1.56 (m, 9H), 1.45 (s, 3H), 1.26-1.04 (m, 7H), 0.96 (s, 3H).
13C NMR (151 MHz, CDCl3): δ 202.8, 186.5, 169.5, 155.8, 128.0, 122.6, 107.7, 97.2, 81.9, 71.6, 69.9, 55.0, 49.7, 46.1, 43.9, 40.9, 40.6, 39.5, 33.9, 33.3, 31.8, 30.3, 27.1, 26.2, 25.5, 21.1, 17.1.
(Cic4の合成 )
Cic3 (265 mg, 0.48 mmol)のアセトニトリル溶液 (2.4 mL)にアジ化ナトリウム (156 mg, 2.4 mmol)を加え、60℃にて48時間攪拌した。反応液を室温に戻した後、酢酸エチルで希釈し、水、飽和食塩水で洗浄、無水硫酸ナトリウム上で乾燥後、ろ過して溶媒を減圧留去した。得られた残渣をフラッシュシリカゲルカラムクロマトグラフィー （ヘキサン/酢酸エチル = 4 : 1 to 1 : 1）にて精製し、Cic4を白色泡状固体として得た (115 mg, 48%)。
1H NMR (600 MHz, CDCl3) : δ 7.25 (d, J = 10.8 Hz, 1H), 6.29 (dd, J = 10.8, 1.8 Hz, 1H), 6.04 (s, 1H), 4.89 (d, J = 4.8 Hz, 1H), 4.52 (t, J = 3.0 Hz, 1H), 4.29 (d, J = 4.8 Hz, 1H), 4.17 (d, J = 18.6 Hz, 1H), 3.92 (d, J = 18.6 Hz, 1H), 2.57 (ddd, J = 12.6, 12.6, 4.8 Hz, 1H), 2.35 (dd, J = 13.2, 3.0 Hz, 1H), 2.18-2.12 (m, 1H), 2.11-2.06 (m, 1H), 1.79-1.54 (m, 12H), 1.46 (s, 3H), 1.26-1.04 (m, 7H), 0.94 (s, 3H).
13C NMR (151 MHz, CDCl3) : δ 204.6, 186.5, 169.6, 155.8, 128.0, 122.6, 107.5, 97.3, 81.7, 70.0, 55.9, 55.1, 49.8, 45.8, 43.9, 41.2, 40.6, 33.9, 33.2, 31.8, 30.3, 27.1, 26.2, 25.5, 21.1, 17.5.
HRMS (ESI) m/z calculated for C28H38N3O5+ 496.2806 [M + H]+ found 496.2802.
(Synthesis of Cic4)
Sodium azide (156 mg, 2.4 mmol) was added to a solution of Cic3 (265 mg, 0.48 mmol) in acetonitrile (2.4 mL) and the mixture was stirred at 60°C for 48 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate, washed with water and saturated saline, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The resulting residue was purified by flash silica gel column chromatography (hexane/ethyl acetate = 4:1 to 1:1) to give Cic4 as a white foamy solid (115 mg, 48%).
1H NMR (600 MHz, CDCl3): δ 7.25 (d, J = 10.8 Hz, 1H), 6.29 (dd, J = 10.8, 1.8 Hz, 1H), 6.04 (s, 1H), 4.89 (d, J = 4.8 Hz, 1H), 4.52 (t, J = 3.0 Hz, 1H), 4.29 (d, J = 4.8 Hz, 1H), 4.17 (d, J = 18.6 Hz, 1H), 3.92 (d, J = 18.6 Hz, 1H), 2.57 (ddd, J = 12.6, 12.6, 4.8 Hz, 1H), 2.35 (dd, J = 13.2, 3.0 Hz, 1H), 2.18-2.12 (m, 1H), 2.11-2.06 (m, 1H), 1.79-1.54 (m, 12H), 1.46 (s, 3H), 1.26-1.04 (m, 7H), 0.94 (s, 3H).
13C NMR (151 MHz, CDCl3): δ 204.6, 186.5, 169.6, 155.8, 128.0, 122.6, 107.5, 97.3, 81.7, 70.0, 55.9, 55.1, 49.8, 45.8, 43.9, 41.2, 40.6, 33.9, 33.2, 31.8, 30.3, 27.1, 26.2, 25.5, 21.1, 17.5.
HRMS (ESI) m/z calculated for C28H38N3O5+ 496.2806 [M + H]+ found 496.2802.
(Cic5の合成 )
Cic4 (12 mg, 0.02 mmol)のテトラヒドロフラン溶液 (267 μL)に1M塩酸 (133 μL)を加えた後、トリフェニルホスフィン (9 mg, 0.03 mmol)を加え、室温にて12時間攪拌した。反応液を減圧乾固した後、残渣をアセトニトリルにて2回共沸した。得られた残渣をジクロロメタン (460 μL)に溶解させ、0℃にてイソ酪酸無水物 (8 μL, 0.046 mol)、次いでトリエチルアミン (13 μL, 0.092 mmol)を加えた。室温で2時間撹拌した後、反応液を減圧濃縮し、得られた残渣をフラッシュシリカゲルカラムクロマトグラフィー （ヘキサン/酢酸エチル = 4 : 1 to 1 : 2）にて精製し、Cic5を白色泡状固体として得た (9.5 mg, 77%)。
1H NMR (600 MHz, CDCl3) : δ 7.27 (d, J = 9.6 Hz, 1H), 6.28 (dd, J = 9.6, 1.8 Hz, 1H), 6.17 (br.t, J = 4.2 Hz, 1H), 6.03 (br.s, 1H), 4.87 (d, J = 6.0 Hz, 1H), 4.50 (d, J = 3.0 Hz, 1H), 4.30 (dd, J = 14.4, 4.2 Hz, 1H), 4.27 (t, J = 4.2 Hz, 1H), 4.12 (dd, J = 14.4, 4.2 Hz, 1H), 2.56 (ddd, J = 13.2, 13.2, 4.8 Hz, 1H), 2.46 (quint., J =7.2 Hz, 1H), 2.34 (dd, J = 13.8, 3.0 Hz, 1H), 2.15 (ddd, J = 10.8, 10.8, 2.4 Hz, 1H), 2.09 (dd, J = 13.8, 2.4 Hz, 1H), 1.93 (dd, J = 13.8, 3.0 Hz, 1H), 1.85 (dd, J = 14.4, 2.4 Hz, 2H), 1.80-1.70 (m, 5H), 1.67-1.53 (m, 4H), 1.45 (s, 3H), 1.26-1.01 (m, 7H), 1.20 (d, J = 7.2 Hz, 3H), 1.18 (d, J = 6.6 Hz, 3H), 0.91 (s, 3H).
13C NMR (151 MHz, CDCl3) : δ 206.2, 186.5, 177.4, 169.6, 155.9, 128.0, 122.6, 107.4, 97.4, 81.6, 69.9, 55.1, 49.8, 47.7, 45.9, 44.0, 40.8, 40.6, 35.4, 34.0, 33.3, 31.9, 30.4, 27.1, 26.2, 25.5, 21.1, 19.5, 19.5, 17.1.
(Synthesis of Cic5)
To a solution (267 μL) of Cic4 (12 mg, 0.02 mmol) in tetrahydrofuran, 1M hydrochloric acid (133 μL) was added, followed by triphenylphosphine (9 mg, 0.03 mmol), and the mixture was stirred at room temperature for 12 hours. The reaction mixture was evaporated to dryness under reduced pressure, and the residue was azeotroped twice with acetonitrile. The resulting residue was dissolved in dichloromethane (460 μL), and isobutyric anhydride (8 μL, 0.046 mol) and triethylamine (13 μL, 0.092 mmol) were added at 0°C. After stirring at room temperature for 2 hours, the reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by flash silica gel column chromatography (hexane/ethyl acetate = 4:1 to 1:2) to give Cic5 as a white foamy solid (9.5 mg, 77%).
1H NMR (600 MHz, CDCl3): δ 7.27 (d, J = 9.6 Hz, 1H), 6.28 (dd, J = 9.6, 1.8 Hz, 1H), 6.17 (br.t, J = 4.2 Hz, 1H), 6.03 (br.s, 1H), 4.87 (d, J = 6.0 Hz, 1H), 4.50 (d, J = 3.0 Hz, 1H), 4.30 (dd, J = 14.4, 4.2 Hz, 1H), 4.27 (t, J = 4.2 Hz, 1H), 4.12 (dd, J = 14.4, 4.2 Hz, 1H), 2.56 (ddd, J = 13.2, 13.2, 4.8 Hz, 1H), 2.46 (quint., J =7.2 Hz, 1H), 2.34 (dd, J = 13.8, 3.0 Hz, 1H), 2.15 (ddd, J = 10.8, 10.8, 2.4 Hz, 1H), 2.09 (dd, J = 13.8, 2.4 Hz, 1H), 1.93 (dd, J = 13.8, 3.0 Hz, 1H), 1.85 (dd, J = 14.4, 2.4 Hz, 2H), 1.80-1.70 (m, 5H), 1.67-1.53 (m, 4H), 1.45 (s, 3H), 1.26-1.01 (m, 7H), 1.20 (d, J = 7.2 Hz, 3H), 1.18 (d, J = 6.6 Hz, 3H), 0.91 (s, 3H).
13C NMR (151 MHz, CDCl3): δ 206.2, 186.5, 177.4, 169.6, 155.9, 128.0, 122.6, 107.4, 97.4, 81.6, 69.9, 55.1, 49.8, 47.7, 45.9, 44.0, 40.8, 40.6, 35.4, 34.0, 33.3, 31.9, 30.4, 27.1, 26.2, 25.5, 21.1, 19.5, 19.5, 17.1.
(Cic6の合成 )
Cic3 (25.0 mg, 0.046 mmol)のN,N-ジメチルホルムアミド溶液 (1 mL)に塩化リチウム (9.7 mg, 0.23 mmol)を加え、60℃にて3時間撹拌した。反応液を室温に戻した後、酢酸エチルで希釈し、水、飽和食塩水で洗浄、無水硫酸ナトリウム上で乾燥後、ろ過して溶媒を減圧留去した。得られた残渣をフラッシュシリカゲルカラムクロマトグラフィー （ヘキサン/酢酸エチル = 3 : 1）にて精製し、Cic6を白色固体として得た (19.2 mg, 86 %)。
1H NMR (600 MHz, CDCl3) : δ 7.26 (d, J = 4.2 Hz, 1H), 6.28 (dd, J = 9.6, 1.8 Hz, 1H), 6.02 (br.s, 1H), 4.89 (d, J = 4.8 Hz, 1H), 4.51 (br.s, 1H), 4.40 (d, J = 16.2 Hz, 1H), 4.28 (d, J = 4.2 Hz, 1H), 4.22 (d, J = 17.4 Hz, 1H), 2.56 (td, J = 8.4, 6.0 Hz, 1H), 2.34 (dd, J =13.8, 3.0 Hz, 1H), 2.16-2.07 (m, 3H), 1.77-1.59 (m, 10H), 1.45 (s, 3H), 1.21-1.08 (m, 8H), 0.92 (s, 3H).
13C NMR (151 MHz, CDCl3) : δ 201.4, 186.5, 169.8, 155.9, 128.0, 122.5, 107.5, 97.8, 81.9, 69.9, 55.1, 49.7, 47.6, 45.6, 44.0, 41.1, 40.6, 33.9, 33.2, 31.8, 30.3, (27.1, 27.1), 26.2, 25.5 (1C overlapped), 21.1, 17.6.
(Synthesis of Cic6)
Lithium chloride (9.7 mg, 0.23 mmol) was added to a solution of Cic3 (25.0 mg, 0.046 mmol) in N,N-dimethylformamide (1 mL) and the mixture was stirred at 60°C for 3 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate, washed with water and saturated saline, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The resulting residue was purified by flash silica gel column chromatography (hexane/ethyl acetate = 3:1) to give Cic6 as a white solid (19.2 mg, 86%).
1H NMR (600 MHz, CDCl3): δ 7.26 (d, J = 4.2 Hz, 1H), 6.28 (dd, J = 9.6, 1.8 Hz, 1H), 6.02 (br.s, 1H), 4.89 (d, J = 4.8 Hz, 1H), 4.51 (br.s, 1H), 4.40 (d, J = 16.2 Hz, 1H), 4.28 (d, J = 4.2 Hz, 1H), 4.22 (d, J = 17.4 Hz, 1H), 2.56 (td, J = 8.4, 6.0 Hz, 1H), 2.34 (dd, J =13.8, 3.0Hz, 1H), 2.16-2.07 (m, 3H), 1.77-1.59 (m, 10H), 1.45 (s, 3H), 1.21-1.08 (m, 8H), 0.92 (s, 3H).
13C NMR (151 MHz, CDCl3): δ 201.4, 186.5, 169.8, 155.9, 128.0, 122.5, 107.5, 97.8, 81.9, 69.9, 55.1, 49.7, 47.6, 45.6, 44.0, 41.1, 40.6, 33.9, 33.2, 31.8, 30.3, (27.1, 27.1), 26.2, 25.5 (1C overlapped), 21.1, 17.6.
(Cic7の合成 )
Cic3 (37.0 mg, 0.067 mmol)のN,N-ジメチルホルムアミド溶液 (1 mL)に臭化リチウム (23.4 mg, 0.27 mmol)を加え、60℃にて1時間撹拌した。反応液を室温に戻した後、酢酸エチルで希釈し、水、飽和食塩水で洗浄、無水硫酸ナトリウム上で乾燥後、ろ過して溶媒を減圧留去した。得られた残渣をフラッシュシリカゲルカラムクロマトグラフィー （ヘキサン/酢酸エチル = 3 : 1）にて精製し、Cic7を白色固体として得た (30 mg, 85 %)。
1H NMR (600 MHz, CDCl3) : δ 7.24 (d, J = 10.2 Hz, 1H), 6.30 (d, J = 1.8 Hz, 1H), 6.03 (br.s, 1H), 4.90 (d, J = 4.8 Hz, 1H), 4.52 (br.s, 1H), 4.32 (d, J = 4.2 Hz, 1H), 4.22 (d, J = 14.4 Hz, 1H), 4.06 (d, J = 14.4 Hz, 1H), 2.56 (td, J = 9.6, 8.4 Hz, 1H), 2.35 (dd, J =10.8, 3.0 Hz, 1H), 2.14-2.04 (m, 3H), 1.77-1.58 (m, 10H), 1.45 (s, 3H), 1.34 (br.s, 1H), 1.20-1.09 (m, 7H), 0.93 (s, 3H).
13C NMR (151 MHz, CDCl3) : δ 201.4, 186.5, 169.6, 155.8, 128.0, 122.6, 107.5, 98.0, 81.9, 70.0, 55.1, 49.6, 45.7, 43.9, 41.3, 40.6, 33.9, 33.3, 33.2, 31.8, 30.3, 27.2, 27.1, 26.2, 25.5 (1C overlapped), 21.1, 17.7.
(Synthesis of Cic7)
Lithium bromide (23.4 mg, 0.27 mmol) was added to a solution of Cic3 (37.0 mg, 0.067 mmol) in N,N-dimethylformamide (1 mL) and stirred at 60°C for 1 hour. The reaction mixture was cooled to room temperature, diluted with ethyl acetate, washed with water and saturated saline, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The resulting residue was purified by flash silica gel column chromatography (hexane/ethyl acetate = 3:1) to give Cic7 as a white solid (30 mg, 85%).
1H NMR (600 MHz, CDCl3): δ 7.24 (d, J = 10.2 Hz, 1H), 6.30 (d, J = 1.8 Hz, 1H), 6.03 (br.s, 1H), 4.90 (d, J = 4.8 Hz, 1H), 4.52 (br.s, 1H), 4.32 (d, J = 4.2 Hz, 1H), 4.22 (d, J = 14.4 Hz, 1H), 4.06 (d, J = 14.4 Hz, 1H), 2.56 (td, J = 9.6, 8.4 Hz, 1H), 2.35 (dd, J =10.8, 3.0 Hz, 1H), 2.14-2.04 (m, 3H), 1.77-1.58 (m, 10H), 1.45 (s, 3H), 1.34 (br.s, 1H), 1.20-1.09 (m, 7H), 0.93 (s, 3H).
13C NMR (151 MHz, CDCl3): δ 201.4, 186.5, 169.6, 155.8, 128.0, 122.6, 107.5, 98.0, 81.9, 70.0, 55.1, 49.6, 45.7, 43.9, 41.3, 40.6, 33.9, 33.3, 33.2, 31.8, 30.3, 27.2, 27.1, 26.2, 25.5 (1C overlapped), 21.1, 17.7.
(Cic8の合成 )
Cic3 (27.4 mg, 0.05 mmol)のN,N-ジメチルホルムアミド溶液 (0.3 mL)にヨウ化ナトリウム (15 mg, 0.10 mmol)を加え、60℃にて1時間撹拌した。反応液を室温に戻した後、酢酸エチルで希釈し、水、飽和食塩水で洗浄、無水硫酸ナトリウム上で乾燥後、ろ過して溶媒を減圧留去した。得られた残渣をフラッシュシリカゲルカラムクロマトグラフィー （ヘキサン/酢酸エチル = 4 : 1 to 1 : 1）にて精製し、Cic8を白色泡状固体として得た (14 mg, 48%)。
1H NMR (600 MHz, CDCl3) : δ 7.26 (d, J = 10.2 Hz, 1H), 6.29 (dd, J = 10.2, 1.8 Hz, 1H), 6.04 (s, 1H), 4.89 (d, J = 5.4 Hz, 1H), 4.53 (br. s, 1H), 4.41 (d, J = 4.8 Hz, 1H), 4.09 (d, J = 11.4 Hz, 1H), 3.92 (d, J = 11.4 Hz, 1H), 2.57 (ddd, J = 13.8, 13.8, 5.4 Hz, 1H), 2.35 (dd, J = 13.2, 3.0 Hz, 1H), 2.18-2.05 (m, 3H), 1.77-1.56 (m, 10H), 1.46 (s, 3H), 1.23-1.05 (m, 7H), 0.93 (s, 3H).
13C NMR (151 MHz, CDCl3) : δ 203.2, 186.5, 169.7, 155.9, 128.0, 122.6, 107.3, 98.1, 81.7, 70.0, 55.1, 49.4, 45.9, 45.6, 43.9, 41.3, 40.7, 33.9, 33.3, 31.8, 30.4, 27.2, 26.2, 25.5, 21.1, 17.9.
(Synthesis of Cic8)
Sodium iodide (15 mg, 0.10 mmol) was added to a solution of Cic3 (27.4 mg, 0.05 mmol) in N,N-dimethylformamide (0.3 mL) and stirred at 60°C for 1 hour. The reaction mixture was cooled to room temperature, diluted with ethyl acetate, washed with water and saturated saline, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The resulting residue was purified by flash silica gel column chromatography (hexane/ethyl acetate = 4:1 to 1:1) to give Cic8 as a white foamy solid (14 mg, 48%).
1H NMR (600 MHz, CDCl3): δ 7.26 (d, J = 10.2 Hz, 1H), 6.29 (dd, J = 10.2, 1.8 Hz, 1H), 6.04 (s, 1H), 4.89 (d, J = 5.4 Hz, 1H), 4.53 (br. s, 1H), 4.41 (d, J = 4.8 Hz, 1H), 4.09 (d, J = 11.4 Hz, 1H), 3.92 (d, J = 11.4 Hz, 1H), 2.57 (ddd, J = 13.8, 13.8, 5.4 Hz, 1H), 2.35 (dd, J = 13.2, 3.0Hz, 1H), 2.18-2.05 (m, 3H), 1.77-1.56 (m, 10H), 1.46 (s, 3H), 1.23-1.05 (m, 7H), 0.93 (s, 3H).
13C NMR (151 MHz, CDCl3): δ 203.2, 186.5, 169.7, 155.9, 128.0, 122.6, 107.3, 98.1, 81.7, 70.0, 55.1, 49.4, 45.9, 45.6, 43.9, 41.3, 40.7, 33.9, 33.3, 31.8, 30.4, 27.2, 26.2, 25.5, 21.1, 17.9.
(Cic9の合成 )
Cic7 (27.4 mg, 0.05 mmol)をジメチルアミン溶液 (2Mテトラヒドロフラン)(400 μL)に溶解させ、60℃にて12時間撹拌した。反応液を減圧留去した後、得られた残渣をフラッシュシリカゲルカラムクロマトグラフィー （ジクロロメタン/メタノール = 100 : 0 to 90 : 10）にて精製し、Cic9を淡黄色油状物質として得た (4.5 mg, 18%)。
1H NMR (600 MHz, CDCl3) : δ 7.23 (d, J = 9.6 Hz, 1H), 6.28 (dd, J = 9.6, 1.8 Hz, 1H), 6.01 (dd, J = 6.0, 1.8 Hz, 1H), 4.86 (d, J = 6.3 Hz, 1H), 4.49 (d, J = 3.0 Hz, 1H), 4.23 (d, J = 4.2 Hz, 1H), 3.67 (t, J = 6.3 Hz, 1H), 3.51 (d, J = 19.2 Hz, 1H), 3.17 (d, J = 19.2 Hz, 1H), 2.54 (ddd, J = 13.8, 13.8, 5.4 Hz, 1H), 2.30 (s, 6H), 2.15-2.04 (m, 2H), 1.75-1.51 (m, 12H), 1.44 (s, 3H), 1.27-1.02 (m, 7H), 0.90 (s, 3H).
13C NMR (151 MHz, CDCl3) : δ 207.2, 186.6, 169.7, 155.8, 128.1, 122.7, 107.2, 97.6, 81.5, 70.3, 63.0, 55.3, 50.0, 45.9, 45.6, 44.1, 41.6, 40.8, 34.1, 33.4, 32.0, 30.5, 27.3, 26.4, 25.7(1C overlapped), 21.3, 17.7.
(Synthesis of Cic9)
Cic7 (27.4 mg, 0.05 mmol) was dissolved in dimethylamine solution (2 M tetrahydrofuran) (400 μL) and stirred for 12 hours at 60° C. The reaction mixture was evaporated under reduced pressure, and the resulting residue was purified by flash silica gel column chromatography (dichloromethane/methanol = 100 : 0 to 90 : 10) to give Cic9 as a pale yellow oil (4.5 mg, 18%).
1H NMR (600 MHz, CDCl3): δ 7.23 (d, J = 9.6 Hz, 1H), 6.28 (dd, J = 9.6, 1.8 Hz, 1H), 6.01 (dd, J = 6.0, 1.8 Hz, 1H), 4.86 (d, J = 6.3 Hz, 1H), 4.49 (d, J = 3.0 Hz, 1H), 4.23 (d, J = 4.2 Hz, 1H), 3.67 (t, J = 6.3 Hz, 1H), 3.51 (d, J = 19.2 Hz, 1H), 3.17 (d, J = 19.2 Hz, 1H), 2.54 (ddd, J = 13.8, 13.8, 5.4 Hz, 1H), 2.30 (s, 6H), 2.15-2.04 (m, 2H), 1.75-1.51 (m, 12H), 1.44 (s, 3H), 1.27-1.02 (m, 7H), 0.90 (s, 3H).
13C NMR (151 MHz, CDCl3): δ 207.2, 186.6, 169.7, 155.8, 128.1, 122.7, 107.2, 97.6, 81.5, 70.3, 63.0, 55.3, 50.0, 45.9, 45.6, 44.1, 41.6, 40.8, 34.1, 33.4, 32.0, 30.5, 27.3, 26.4, 25.7(1C overlapped), 21.3, 17.7.
(Cic10の合成 )
Cic4 (24.8 mg, 0.05 mmol)のテトラヒドロフラン溶液 (580 μL)に1M塩酸 (290 μL)を加えた後、トリフェニルホスフィン (20 mg, 0.075 mmol)を加え、室温にて12時間撹拌した。反応液を減圧乾固した後、残渣を水 (2 mL)に懸濁させ、ジクロロメタン (1 mL x 2)で洗浄した。水層を真空下にて濃縮し、白色固体を得た (22 mg, 85%)。
1H NMR (600 MHz, MeOH-d4) : δ 7.44 (d, J = 10.2 Hz, 1H), 6.25 (dd, J = 10.2, 1.8 Hz, 1H), 6.01 (s, 1H), 4.44 (dd, J = 6.6, 3.0 Hz, 1H), 4.38 (d, J = 4.8 Hz, 1H), 4.17 (d, J = 18.6 Hz, 1H), 3.85 (d, J = 18.6 Hz, 1H), 2.64 (ddd, J = 13.2, 13.2, 1.5 Hz, 1H), 2.37 (dd, J = 13.2, 3.0 Hz, 1H), 2.26-2.19 (m, 1H), 2.15-2.11 (m, 1H), 1.98 (dd, J = 13.2, 3.6 Hz, 1H), 1.80-1.72 (m, 6H), 1.68-1.56 (m, 4H), 1.48 (s, 3H), 1.27-0.99 (m, 8H), 0.97 (s, 3H).
13C NMR (151 MHz, MeOH-d4) : δ 204.5, 188.8, 174.1, 159.5, 128.0, 122.7, 108.6, 98.6, 83.2, 70.3, 57.2, 51.4, 47.5, 47.5, 45.9, 42.1, 41.4, 35.5, 34.3, 33.0, 31.7, 28.3, 27.4, 26.7, 21.5, 17.6.
(Synthesis of Cic10)
To a solution of Cic4 (24.8 mg, 0.05 mmol) in tetrahydrofuran (580 μL), 1M hydrochloric acid (290 μL) was added, followed by triphenylphosphine (20 mg, 0.075 mmol) and stirring at room temperature for 12 hours. The reaction mixture was evaporated to dryness under reduced pressure, and the residue was suspended in water (2 mL) and washed with dichloromethane (1 mL x 2). The aqueous layer was concentrated in vacuum to give a white solid (22 mg, 85%).
1H NMR (600 MHz, MeOH-d4): δ 7.44 (d, J = 10.2 Hz, 1H), 6.25 (dd, J = 10.2, 1.8 Hz, 1H), 6.01 (s, 1H), 4.44 (dd, J = 6.6, 3.0 Hz, 1H), 4.38 (d, J = 4.8 Hz, 1H), 4.17 (d, J = 18.6 Hz, 1H), 3.85 (d, J = 18.6 Hz, 1H), 2.64 (ddd, J = 13.2, 13.2, 1.5 Hz, 1H), 2.37 (dd, J = 13.2, 3.0 Hz, 1H), 2.26-2.19 (m, 1H), 2.15-2.11 (m, 1H), 1.98 (dd, J = 13.2, 3.6 Hz, 1H), 1.80-1.72 (m, 6H), 1.68-1.56 (m, 4H), 1.48 (s, 3H), 1.27-0.99 (m, 8H), 0.97 (s, 3H).
13C NMR (151 MHz, MeOH-d4): δ 204.5, 188.8, 174.1, 159.5, 128.0, 122.7, 108.6, 98.6, 83.2, 70.3, 57.2, 51.4, 47.5, 47.5, 45.9, 42.1, 41.4, 35.5, 34.3, 33.0, 31.7, 28.3, 27.4, 26.7, 21.5, 17.6.
(Cic11-1,11-2の合成 )
Cic3 (89.9 mg, 0.164 mmol)をN,N-ジメチルホルムアミド(1 mL)に溶解させ、室温にて24時間撹拌した。酢酸エチルで希釈し、水、飽和食塩水で洗浄、無水硫酸ナトリウム上で乾燥後、ろ過して溶媒を減圧留去した。得られた残渣をフラッシュシリカゲルカラムクロマトグラフィー （ヘキサン/酢酸エチル = 3 : 1 to 1 : 1）にて精製し、Cic11-1，Cic11-2を白色固体として得た (Cic11-1, 19.9 mg, 25%)(Cic11-2, 8.4 mg, 11%)。
Cic11-1
1H NMR (600 MHz, CDCl3) : δ 7.29 (d, J = 10.2 Hz, 1H), 6.30 (dd, J = 10.2, 1.8 Hz, 1H), 6.04 (s, 1H), 4.68 (d, J = 4.2 Hz, 1H), 4.51 (br.s, 1H), 4.17 (d, J = 5.4 Hz, 1H), 3.21 (d, J = 4.8 Hz, 1H), 3.16 (d, J = 4.8 Hz, 1H), 2.57 (td, J = 8.4, 6.0 Hz, 1H), 2.34 (dd, J = 12.9, 2.4 Hz, 1H), 2.18-2.15 (m, 2H), 2.04-2.00 (m, 1H), 1.75-1.61 (m, 9H), 1.52-1.47 (s, 5H), 1.27-1.01 (m, 10H).
13C NMR (151 MHz, CDCl3) : δ 186.6, 169.6, 156.0, 128.0, 122.6, 108.4, 91.0, 79.3, 70.0, 55.1, 49.2, 49.0, 46.9, 46.5, 44.0, 41.4, 40.9, 33.9, 33.2, 31.8, 31.5, 30.1, 27.3, 27.1, 26.2, 25.6 (1C overlapped), 21.1, 16.6.
Cic11-2
1H NMR (600 MHz, CDCl3) : δ 7.27 (d, J = 10.2 Hz, 1H), 6.29 (dd, J = 9.9, 1.8 Hz, 1H), 6.03 (s, 1H), 4.70 (d, J = 3.6 Hz, 1H), 4.48 (d, J = 3.0 Hz, 1H), 4.25 (d, J = 6.0 Hz, 1H), 3.27 (d, J = 4.8 Hz, 1H), 3.01 (d, J = 4.8 Hz, 1H), 2.56 (td, J = 13.2, 8.4 Hz, 1H), 2.34 (dd, J = 16.2, 3.0 Hz, 1H), 2.15-2.00 (m, 3H), 1.81-1.41 (m, 14H), 1.27-1.05 (m, 10H).
13C NMR (151 MHz, CDCl3) : δ 186.5, 169.5, 155.8, 128.0, 122.6, 107.9, 91.4, 79.8, 70.0, 55.1, 50.2, 50.0, 48.9, 46.7, 44.0, 41.0, 40.8, 33.9, 33.4, 31.8, 30.1, 27.2, 27.0, 26.2, 25.6 (1C overlapped), 21.1, 17.3.
(Synthesis of Cic11-1 and Cic11-2)
Cic3 (89.9 mg, 0.164 mmol) was dissolved in N,N-dimethylformamide (1 mL) and stirred at room temperature for 24 h. The mixture was diluted with ethyl acetate, washed with water and saturated saline, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The residue was purified by flash silica gel column chromatography (hexane/ethyl acetate = 3:1 to 1:1) to give Cic11-1 and Cic11-2 as white solids (Cic11-1, 19.9 mg, 25%) (Cic11-2, 8.4 mg, 11%).
Cic11-1
1H NMR (600 MHz, CDCl3): δ 7.29 (d, J = 10.2 Hz, 1H), 6.30 (dd, J = 10.2, 1.8 Hz, 1H), 6.04 (s, 1H), 4.68 (d, J = 4.2 Hz, 1H), 4.51 (br.s, 1H), 4.17 (d, J = 5.4 Hz, 1H), 3.21 (d, J = 4.8 Hz, 1H), 3.16 (d, J = 4.8 Hz, 1H), 2.57 (td, J = 8.4, 6.0 Hz, 1H), 2.34 (dd, J = 12.9, 2.4 Hz, 1H), 2.18-2.15 (m, 2H), 2.04-2.00 (m, 1H), 1.75-1.61 (m, 9H), 1.52-1.47 (s, 5H), 1.27-1.01 (m, 10H).
13C NMR (151 MHz, CDCl3): δ 186.6, 169.6, 156.0, 128.0, 122.6, 108.4, 91.0, 79.3, 70.0, 55.1, 49.2, 49.0, 46.9, 46.5, 44.0, 41.4, 40.9, 33.9, 33.2, 31.8, 31.5, 30.1, 27.3, 27.1, 26.2, 25.6 (1C overlapped), 21.1, 16.6.
Cic11-2
1H NMR (600 MHz, CDCl3): δ 7.27 (d, J = 10.2 Hz, 1H), 6.29 (dd, J = 9.9, 1.8 Hz, 1H), 6.03 (s, 1H), 4.70 (d, J = 3.6 Hz, 1H), 4.48 (d, J = 3.0 Hz, 1H), 4.25 (d, J = 6.0 Hz, 1H), 3.27 (d, J = 4.8 Hz, 1H), 3.01 (d, J = 4.8 Hz, 1H), 2.56 (td, J = 13.2, 8.4 Hz, 1H), 2.34 (dd, J = 16.2, 3.0 Hz, 1H), 2.15-2.00 (m, 3H), 1.81-1.41 (m, 14H), 1.27-1.05 (m, 10H).
13C NMR (151 MHz, CDCl3): δ 186.5, 169.5, 155.8, 128.0, 122.6, 107.9, 91.4, 79.8, 70.0, 55.1, 50.2, 50.0, 48.9, 46.7, 44.0, 41.0, 40.8, 33.9, 33.4, 31.8, 30.1, 27.2, 27.0, 26.2, 25.6 (1C overlapped), 21.1, 17.3.
(3)毒性試験
Vero E6/TMPRSS2（JCRB 1819）細胞を、異なる終濃度（2.5μM、5μM、10μM、20μM、40μM）の被験化合物を含む培地（2%FCS、ペニシリン（100 units/mL）、ストレプトマイシン（100μg/mL）加DMEM）を用いて18時間培養した。Cytotoxicity LDH Assay Kit-WST（同仁化学研究所）を用いて各被験化合物存在下における細胞傷害性を評価した。数値化にあたっては、Nivoマルチモードマイクロプレートリーダー（PerkinElmer）を用いて480nmの吸光度を測定した。各被験化合物、各濃度につき、N=3とした。対照群にはDMSOを用いた。
(3) Toxicity testing
Vero E6/TMPRSS2 (JCRB 1819) cells were cultured for 18 hours in medium (DMEM supplemented with 2% FCS, penicillin (100 units/mL), and streptomycin (100 μg/mL)) containing test compounds at different final concentrations (2.5 μM, 5 μM, 10 μM, 20 μM, and 40 μM). Cytotoxicity in the presence of each test compound was evaluated using the Cytotoxicity LDH Assay Kit-WST (Dojindo Laboratories). For quantification, absorbance at 480 nm was measured using a Nivo multimode microplate reader (PerkinElmer). N=3 was used for each test compound and each concentration. DMSO was used as a control.
異なる濃度の被験化合物をVero E6/TMPRSS2 細胞に18時間共在させた際の細胞毒性について、LDHを指標とした細胞毒性試験により評価した。結果として、シクレソニド（Cicle）を含む計10化合物のうち、シクレソニド 代謝物であるCic2、Cic3、Cic7、Cic8、Cic9、Cic10、Cic11-2の7化合物は40μM濃度で細胞毒性を示した一方で、4化合物（Cicle、Cic4、Cic6、Cic11-1）は40μM濃度においてもLDH放出は10%未満となり、Cic4、Cic6、Cic11-1はCicleと同等の細胞毒性に留まることが示された（図２）。 The cytotoxicity of the test compounds was evaluated by a cytotoxicity test using LDH as an index when they were coexisted with Vero E6/TMPRSS2 cells at different concentrations for 18 hours. As a result, out of a total of 10 compounds including ciclesonide (Cicle), seven compounds, which are metabolites of ciclesonide, Cic2, Cic3, Cic7, Cic8, Cic9, Cic10, and Cic11-2, showed cytotoxicity at a concentration of 40 μM, while four compounds (Cicle, Cic4, Cic6, and Cic11-1) showed less than 10% LDH release even at a concentration of 40 μM, indicating that Cic4, Cic6, and Cic11-1 have the same cytotoxicity as Cicle (Figure 2).
(4)ウイルス感染実験
SARS-CoV-2 JPN/TY/WK-521株の培養には、上述のVero E6/TMPRSS2細胞を用いた。同細胞株は、5%熱不活化ウシ胎児血清（FBS；SAFC Biosciences, Lenexa, KS, USA）、ペニシリン（100 units/mL）、ストレプトマイシン（100μg/mL）を含むDulbecco’s modified Eagle’s medium（DMEM, low glucose；富士フィルム和光純薬工業）を用い、37℃、5％CO2下で培養した。
(4) Viral infection experiments
The SARS-CoV-2 JPN/TY/WK-521 strain was cultured in Vero E6/TMPRSS2 cells (described above) at 37°C in 5% CO2 in Dulbecco's modified Eagle's medium (DMEM, low glucose; Fujifilm Wako Pure Chemical Industries, Ltd.) containing 5% heat-inactivated fetal bovine serum (FBS; SAFC Biosciences, Lenexa, KS, USA), penicillin (100 units/mL), and streptomycin (100 μg/mL).
ウイルス感染実験に際しては、Vero E6/TMPRSS2細胞を96ウェル平底組織培養用プレート（TPP）に1ウェルあたり1.5 x 104細胞となるように接種し、20時間培養後、上清を吸引除去し、速やかに被験化合物を含む培地（2%FBS、ペニシリン（100units/mL）、ストレプトマイシン（100μg/mL）加DMEM）を1ウェルにつき100μL添加した（各被験化合物、各濃度につき、3ウェルずつ用いた）。その後、2%FBS、ペニシリン（100units/mL）、ストレプトマイシン（100μg/mL）加DMEM を用いて、MOI（Multiplicity of Infection）が0.1となるよう濃度調整した、SARS-CoV-2懸濁液100μLを各ウェルに添加した（被験物質の終濃度は、0.625μM、1.25μM、2.5μM、5μM、10μM、20μM、40μM、80μM）。37℃、5％CO2下で18時間培養後、培養上清全量を回収し、遠心分離後、以下のウイルス力価試験に用いた。同時に上清除去後の細胞からもRNA抽出を行った。対照群にはDMSOを用いた。 For the virus infection experiment, Vero E6/TMPRSS2 cells were seeded into a 96-well flat-bottom tissue culture plate (TPP) at 1.5 x 104 cells per well. After 20 hours of culture, the supernatant was removed by aspiration, and 100 μL of medium containing the test compound (DMEM supplemented with 2% FBS, penicillin (100 units/mL), and streptomycin (100 μg/mL)) was immediately added per well (three wells were used for each test compound and each concentration). Then, 100 μL of SARS-CoV-2 suspension was added to each well, with the concentration adjusted to a multiplicity of infection (MOI) of 0.1 using DMEM supplemented with 2% FBS, penicillin (100 units/mL), and streptomycin (100 μg/mL) (final concentrations of test substances: 0.625 μM, 1.25 μM, 2.5 μM, 5 μM, 10 μM, 20 μM, 40 μM, and 80 μM). After culturing for 18 hours at 37 °C and 5% CO 2 , the entire culture supernatant was collected, centrifuged, and used in the following virus titer test. At the same time, RNA was extracted from the cells after the supernatant was removed. DMSO was used as a control.
細胞培養上清100μLより、Maxwell RSC Viral Total Nucleic Acid Purification Kit（Promega）を用いてRNA抽出を行った。上清除去後の細胞からのRNA抽出には、CellAmpTM Direct RNA Prep Kit for RT-PCR (Real Time)（タカラバイオ）を用いた。 RNA was extracted from 100 μL of cell culture supernatant using Maxwell RSC Viral Total Nucleic Acid Purification Kit (Promega). After removing the supernatant, RNA was extracted from the cells using CellAmp TM Direct RNA Prep Kit for RT-PCR (Real Time) (Takara Bio).
RNA抽出液2.5μLを鋳型として、マスターミックスにはTaqMan(登録商標) Fast Virus 1-Step Master Mix（Thermo Fisher Scientific）を、サーマルサイクラーにはABI7500（Thermo Fisher Scientific）を用いてリアルタイムPCRを行った。プライマー・プローブ、反応液組成及び反応条件は「国立感染症研究所病原体検出マニュアル2019-nCoV Ver.2.6」のN2セットに従った。ウイルスRNA定量にあたっては、2019-nCoV_N_Positive Control（IDT）を用いて予め検量線を作成し、これを用いた絶対定量により各RNA試料中のウイルスRNAコピー数を求めた。反応時の陽性対照には上述の2019-nCoV_N_Positive Controlを、陰性対照にはDMSO投与群より抽出したRNAを用いた。 Real-time PCR was performed using 2.5 μL of RNA extract as a template, TaqMan® Fast Virus 1-Step Master Mix (Thermo Fisher Scientific) as the master mix, and ABI7500 (Thermo Fisher Scientific) as the thermal cycler. The primers and probes, reaction solution composition, and reaction conditions were in accordance with the N2 set of the National Institute of Infectious Diseases Pathogen Detection Manual 2019-nCoV Ver. 2.6. For viral RNA quantification, a calibration curve was created in advance using 2019-nCoV_N_Positive Control (IDT), and the number of viral RNA copies in each RNA sample was determined by absolute quantification using this. The above-mentioned 2019-nCoV_N_Positive Control was used as the positive control during the reaction, and RNA extracted from the DMSO-administered group was used as the negative control.
12ウェル組織培養用プレート（TPP）に2%FBS、ペニシリン（100 units/mL）、ストレプトマイシン（100μg/mL）加DMEM を用いてVeroE6/TMPRSS2細胞を播種し、37℃、5％CO2下で18時間培養した。感染細胞上清検体を2%FBS、ペニシリン（100 units/mL）、ストレプトマイシン（100μg/mL）加DMEMで10倍階段希釈した後、1ウェルあたり50μLの各希釈液を加え、37℃、5%CO2下で1時間保温した。その後、上清を吸引除去し、1%メチルセルロース、2%FBS、ペニシリン（100 units/mL）、ストレプトマイシン（100μg/mL）を含むDMEMを加え、37℃、5%CO2下で72～96時間培養した。培養後は10%中性緩衝ホルマリン液で固定を行い、0.05%メチレンブルー溶液0.5mlを加え、1時間染色後、1ウェルあたりのプラーク数を計数し、試料溶液1ｍLあたりのプラーク数を求めた。各被験物質・濃度につき、N=3を供試し、得られた数値を基に、XLfitプログラム（IDBS）を用いてシグモイド曲線を作成し、初発ウイルス力価を1/100、1/1000に低減する上で必要となる濃度を算出した。 VeroE6/TMPRSS2 cells were seeded in 12-well tissue culture plates (TPP) using DMEM supplemented with 2% FBS, penicillin (100 units/mL), and streptomycin (100 μg/mL) and cultured at 37°C under 5% CO2 for 18 hours. The infected cell supernatant samples were serially diluted 10-fold with DMEM supplemented with 2% FBS, penicillin (100 units/mL), and streptomycin (100 μg/mL), and 50 μL of each dilution was added per well and incubated at 37°C under 5% CO2 for 1 hour. The supernatant was then aspirated and DMEM containing 1% methylcellulose, 2% FBS, penicillin (100 units/mL), and streptomycin (100 μg/mL) was added and cultured at 37°C under 5% CO2 for 72 to 96 hours. After incubation, the wells were fixed with 10% neutral buffered formalin, 0.5 ml of 0.05% methylene blue solution was added, and stained for 1 hour. The number of plaques per well was counted, and the number of plaques per mL of sample solution was calculated. N=3 was used for each test substance and concentration, and a sigmoid curve was created using the XLfit program (IDBS) based on the obtained values, and the concentrations required to reduce the initial virus titer to 1/100 and 1/1000 were calculated.
10μM濃度の被験化合物を含む培地中でSARS-CoV-2 JPN/TY/WK-521株を18時間感染させた際の、上清中のウイルスRNA量をリアルタイムPCRにより評価した。結果として、対照群（No treatment；化合物非存在下）のウイルスRNA量は、4.8x1010copy/wellであったのに対し、Cicle投与群は5.2x108copy/wellと有意な低減を示した。Cicleと同等の細胞毒性を示した化合物のうち、Cic4投与群におけるウイルスRNA量は2.1x105copy/well、Cic6投与群では4.6x105copy/wellと共に、Cicle投与群に比べて顕著なウイルス量の低減を示した（図３）。一方、Cic11-1投与群のウイルスRNA量は1.3x109copy/wellとなり、Cic投与群に比べ、有意な低減は認められなかった（図３）。このほか、Cicle投与群との間で有意なウイルスRNA量の低減を認めた化合物としては、Cic2、Cic3、Cic7が挙げられたが、図３の細胞毒性がCicle投与群よりも有意に高い結果を得たため、以降の評価対象からは除外した。 The viral RNA amount in the supernatant was evaluated by real-time PCR when SARS-CoV-2 JPN/TY/WK-521 strain was infected for 18 hours in a medium containing a test compound at a concentration of 10 μM. As a result, the viral RNA amount in the control group (no treatment; no compound) was 4.8x10 10 copies/well, while the Cicle-treated group showed a significant reduction of 5.2x10 8 copies/well. Among the compounds that showed the same cytotoxicity as Cicle, the viral RNA amount in the Cic4-treated group was 2.1x10 5 copies/well, and the Cic6-treated group was 4.6x10 5 copies/well, showing a significant reduction in the viral amount compared to the Cicle-treated group (Figure 3). On the other hand, the viral RNA amount in the Cic11-1-treated group was 1.3x10 9 copies/well, which was not significantly reduced compared to the Cic-treated group (Figure 3). Other compounds that showed a significant reduction in viral RNA levels compared to the Cicle-administered group were Cic2, Cic3, and Cic7. However, since their cytotoxicity in Figure 3 was significantly higher than that of the Cicle-administered group, they were excluded from further evaluation.
Cic4、Cic6投与によるウイルス増殖阻害効果を更に評価するため、異なる濃度の被験化合物を含む培地中でSARS-CoV-2 JPN/TY/WK-521株を18時間感染後、細胞上清中におけるウイルス力価をプラークアッセイにより評価した。図４の通り、Cicle、Cic2、Cic4、Cic6は何れも濃度依存的にウイルス増殖阻害効果を示し、対照群（DMSO）において認められたウイルス力価（約4.6x105PFU/mL）を1/100または1/1000に低減するために必要となる濃度は、Cicleが5.86μM、＞80.00μM、Cic2が24.55μM、60.95μMであったのに対し、Cic4では6.51μM、7.36μM、Cic6では5.95μM、6.50μMとなる等、Cic4及びCic6はCicle及びCic2に比べ、より高いウイルス増殖阻害効果を有することが示された。 To further evaluate the inhibitory effect of Cic4 and Cic6 on viral proliferation, cells were infected with SARS-CoV-2 JPN/TY/WK-521 strain in medium containing different concentrations of the test compounds for 18 hours, and the viral titer in the cell supernatant was evaluated by plaque assay. As shown in Figure 4, Cicle, Cic2, Cic4, and Cic6 all showed a concentration-dependent inhibitory effect on viral proliferation, and the concentrations required to reduce the viral titer (approximately 4.6x105 PFU/mL) observed in the control group (DMSO) to 1/100 or 1/1000 were 5.86 μM and >80.00 μM for Cicle, 24.55 μM and 60.95 μM for Cic2, while the concentrations were 6.51 μM and 7.36 μM for Cic4, and 5.95 μM and 6.50 μM for Cic6, indicating that Cic4 and Cic6 have a greater inhibitory effect on viral proliferation than Cicle and Cic2.
新型コロナウイルスの予防及び／又は治療に利用できる。 It can be used to prevent and/or treat the new coronavirus.
Claims (16)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2020195723A JP7656784B2 (en) | 2020-11-26 | 2020-11-26 | Novel compounds and anti-coronavirus agents |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2020195723A JP7656784B2 (en) | 2020-11-26 | 2020-11-26 | Novel compounds and anti-coronavirus agents |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2022084098A JP2022084098A (en) | 2022-06-07 |
| JP7656784B2 true JP7656784B2 (en) | 2025-04-04 |
Family
ID=81868263
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2020195723A Active JP7656784B2 (en) | 2020-11-26 | 2020-11-26 | Novel compounds and anti-coronavirus agents |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP7656784B2 (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008546694A (en) | 2005-06-14 | 2008-12-25 | コラス・フアーマ・インコーポレイテツド | Substituted phenyl phosphates as mutual prodrugs of steroids and beta-agonists for the treatment of pneumonia and bronchi |
| JP2015509493A (en) | 2012-02-23 | 2015-03-30 | ベーリンガー インゲルハイム インターナショナル ゲゼルシャフト ミット ベシュレンクテル ハフツング | New method for producing ciclesonide |
-
2020
- 2020-11-26 JP JP2020195723A patent/JP7656784B2/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008546694A (en) | 2005-06-14 | 2008-12-25 | コラス・フアーマ・インコーポレイテツド | Substituted phenyl phosphates as mutual prodrugs of steroids and beta-agonists for the treatment of pneumonia and bronchi |
| JP2015509493A (en) | 2012-02-23 | 2015-03-30 | ベーリンガー インゲルハイム インターナショナル ゲゼルシャフト ミット ベシュレンクテル ハフツング | New method for producing ciclesonide |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2022084098A (en) | 2022-06-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| TWI789695B (en) | Methods for treating sars cov-2 infections | |
| CN116370479B (en) | Use of compound ATV014 in the preparation of products for the treatment of new coronavirus infection | |
| Petit et al. | Docking and in silico toxicity assessment of Arthrospira compounds as potential antiviral agents against SARS-CoV-2 | |
| Lin et al. | Inhibitory effects of some derivatives of glycyrrhizic acid against Epstein-Barr virus infection: Structure–activity relationships | |
| JP2023526179A (en) | Antiviral use of nucleoside analogues or combination preparations containing nucleoside analogues | |
| JP2020090536A (en) | Methods for treating filoviridae virus infections | |
| KR20190112051A (en) | How to Treat Influenza | |
| Wu et al. | Structure properties and mechanisms of action of naturally originated phenolic acids and their derivatives against human viral infections | |
| JP2023516628A (en) | Use of Nucleotide-Based Compounds to Treat Coronavirus Infections | |
| US12358897B2 (en) | Compounds and method of treating COVID-19 | |
| CN113289018A (en) | Application of old medicine such as auranofin and composition thereof in resisting single positive strand RNA virus | |
| JP2023535204A (en) | Application of cannabidiol in treating coronavirus infections | |
| CN113350330B (en) | Application of myricetin compounds in the preparation of drugs for the prevention and treatment of new coronary pneumonia | |
| CN114181258A (en) | Nucleoside compounds for antiviral therapy and uses | |
| Singh et al. | Pongamia pinnata L. seed-derived karanjin as prominent antiviral agent against Newcastle disease virus | |
| JP7656784B2 (en) | Novel compounds and anti-coronavirus agents | |
| Ju et al. | Synthesis and in vitro anti-HSV-1 activity of a novel Hsp90 inhibitor BJ-B11 | |
| CA3219273A1 (en) | Use of 5-nitro-8-hydroxyquinoline | |
| AU2022231012A1 (en) | Suppression of covid-19 replication by covid-19 entry inhibitors | |
| EP4190333A1 (en) | Azelastine as antiviral treatment | |
| CN108619123A (en) | Tenovin-1 is preparing the application in preventing nerpes vinrus hominis's infection medicine | |
| JP2023514794A (en) | Anticoronavirus application of poly ADP ribose polymerase inhibitors | |
| US8044098B2 (en) | Use of hydroxybenzoic acids and their esters and analogues for preventing or treating virus infection | |
| CN116617210A (en) | Application of a small molecular compound in antiviral infection | |
| CN101829103B (en) | Application of flavonoid quercetin dimmer as medicament for treating viral hepatitis B |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| RD01 | Notification of change of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7426 Effective date: 20201210 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20201210 |
|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20230908 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20241112 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20241114 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20250212 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 7656784 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |