JP4716751B2 - Novel compound and method for detecting behavior of bisphenol A using the same - Google Patents
Novel compound and method for detecting behavior of bisphenol A using the same Download PDFInfo
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- JP4716751B2 JP4716751B2 JP2005043187A JP2005043187A JP4716751B2 JP 4716751 B2 JP4716751 B2 JP 4716751B2 JP 2005043187 A JP2005043187 A JP 2005043187A JP 2005043187 A JP2005043187 A JP 2005043187A JP 4716751 B2 JP4716751 B2 JP 4716751B2
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- bisphenol
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- 150000001875 compounds Chemical class 0.000 title claims description 71
- IISBACLAFKSPIT-UHFFFAOYSA-N bisphenol A Chemical compound C=1C=C(O)C=CC=1C(C)(C)C1=CC=C(O)C=C1 IISBACLAFKSPIT-UHFFFAOYSA-N 0.000 title claims description 65
- 238000000034 method Methods 0.000 title claims description 13
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 12
- 239000003153 chemical reaction reagent Substances 0.000 claims description 9
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 6
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 6
- 125000004432 carbon atom Chemical group C* 0.000 claims description 5
- 102000004169 proteins and genes Human genes 0.000 claims description 4
- 108090000623 proteins and genes Proteins 0.000 claims description 4
- 125000000217 alkyl group Chemical group 0.000 claims description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 57
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 46
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 41
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 36
- 239000000243 solution Substances 0.000 description 28
- 239000011541 reaction mixture Substances 0.000 description 27
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 26
- 239000000203 mixture Substances 0.000 description 25
- 238000002360 preparation method Methods 0.000 description 23
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 22
- 239000012300 argon atmosphere Substances 0.000 description 21
- 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 20
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 18
- 235000019341 magnesium sulphate Nutrition 0.000 description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 18
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 17
- 239000012044 organic layer Substances 0.000 description 16
- 239000012153 distilled water Substances 0.000 description 14
- 238000010898 silica gel chromatography Methods 0.000 description 14
- 102000003951 Erythropoietin Human genes 0.000 description 13
- 108090000394 Erythropoietin Proteins 0.000 description 13
- ATUOYWHBWRKTHZ-UHFFFAOYSA-N Propane Chemical compound CCC ATUOYWHBWRKTHZ-UHFFFAOYSA-N 0.000 description 13
- 229940105423 erythropoietin Drugs 0.000 description 13
- OXCMYAYHXIHQOA-UHFFFAOYSA-N potassium;[2-butyl-5-chloro-3-[[4-[2-(1,2,4-triaza-3-azanidacyclopenta-1,4-dien-5-yl)phenyl]phenyl]methyl]imidazol-4-yl]methanol Chemical compound [K+].CCCCC1=NC(Cl)=C(CO)N1CC1=CC=C(C=2C(=CC=CC=2)C2=N[N-]N=N2)C=C1 OXCMYAYHXIHQOA-UHFFFAOYSA-N 0.000 description 13
- 239000011780 sodium chloride Substances 0.000 description 13
- DCXIMRSHMXAVHX-UHFFFAOYSA-N 3-phenyl-3h-diazirine Chemical compound C1=CC=CC=C1C1N=N1 DCXIMRSHMXAVHX-UHFFFAOYSA-N 0.000 description 12
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 12
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 12
- 238000000746 purification Methods 0.000 description 12
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 11
- 210000004027 cell Anatomy 0.000 description 11
- 238000006243 chemical reaction Methods 0.000 description 11
- IJDNQMDRQITEOD-UHFFFAOYSA-N n-butane Chemical compound CCCC IJDNQMDRQITEOD-UHFFFAOYSA-N 0.000 description 10
- 239000011734 sodium Substances 0.000 description 10
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 9
- 238000005222 photoaffinity labeling Methods 0.000 description 9
- ZFFMLCVRJBZUDZ-UHFFFAOYSA-N 2,3-dimethylbutane Chemical group CC(C)C(C)C ZFFMLCVRJBZUDZ-UHFFFAOYSA-N 0.000 description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 8
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 8
- 230000007613 environmental effect Effects 0.000 description 8
- FPGGTKZVZWFYPV-UHFFFAOYSA-M tetrabutylammonium fluoride Chemical compound [F-].CCCC[N+](CCCC)(CCCC)CCCC FPGGTKZVZWFYPV-UHFFFAOYSA-M 0.000 description 8
- 206010021143 Hypoxia Diseases 0.000 description 7
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 7
- 239000007850 fluorescent dye Substances 0.000 description 7
- 238000001215 fluorescent labelling Methods 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- 238000001816 cooling Methods 0.000 description 6
- 230000001146 hypoxic effect Effects 0.000 description 6
- -1 p-hydroxyphenyl Chemical group 0.000 description 6
- 125000000951 phenoxy group Chemical group [H]C1=C([H])C([H])=C(O*)C([H])=C1[H] 0.000 description 6
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 6
- ODLMAHJVESYWTB-UHFFFAOYSA-N propylbenzene Chemical compound CCCC1=CC=CC=C1 ODLMAHJVESYWTB-UHFFFAOYSA-N 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- VPWNQTHUCYMVMZ-UHFFFAOYSA-N 4,4'-sulfonyldiphenol Chemical class C1=CC(O)=CC=C1S(=O)(=O)C1=CC=C(O)C=C1 VPWNQTHUCYMVMZ-UHFFFAOYSA-N 0.000 description 5
- VYSKOVLQUUGJOD-UHFFFAOYSA-N 4-tri(propan-2-yl)silyloxybutan-1-ol Chemical compound CC(C)[Si](C(C)C)(C(C)C)OCCCCO VYSKOVLQUUGJOD-UHFFFAOYSA-N 0.000 description 5
- 229930185605 Bisphenol Natural products 0.000 description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 5
- 239000001273 butane Substances 0.000 description 5
- 238000004587 chromatography analysis Methods 0.000 description 5
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 5
- OFBQJSOFQDEBGM-UHFFFAOYSA-N n-pentane Natural products CCCCC OFBQJSOFQDEBGM-UHFFFAOYSA-N 0.000 description 5
- 239000001301 oxygen Substances 0.000 description 5
- 229910052760 oxygen Inorganic materials 0.000 description 5
- 229920006395 saturated elastomer Polymers 0.000 description 5
- 102000007469 Actins Human genes 0.000 description 4
- 108010085238 Actins Proteins 0.000 description 4
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 4
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 108091081021 Sense strand Proteins 0.000 description 4
- 230000000692 anti-sense effect Effects 0.000 description 4
- KQIADDMXRMTWHZ-UHFFFAOYSA-N chloro-tri(propan-2-yl)silane Chemical compound CC(C)[Si](Cl)(C(C)C)C(C)C KQIADDMXRMTWHZ-UHFFFAOYSA-N 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 4
- 239000001294 propane Substances 0.000 description 4
- WWTBZEKOSBFBEM-SPWPXUSOSA-N (2s)-2-[[2-benzyl-3-[hydroxy-[(1r)-2-phenyl-1-(phenylmethoxycarbonylamino)ethyl]phosphoryl]propanoyl]amino]-3-(1h-indol-3-yl)propanoic acid Chemical compound N([C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)O)C(=O)C(CP(O)(=O)[C@H](CC=1C=CC=CC=1)NC(=O)OCC=1C=CC=CC=1)CC1=CC=CC=C1 WWTBZEKOSBFBEM-SPWPXUSOSA-N 0.000 description 3
- UNILWMWFPHPYOR-KXEYIPSPSA-M 1-[6-[2-[3-[3-[3-[2-[2-[3-[[2-[2-[[(2r)-1-[[2-[[(2r)-1-[3-[2-[2-[3-[[2-(2-amino-2-oxoethoxy)acetyl]amino]propoxy]ethoxy]ethoxy]propylamino]-3-hydroxy-1-oxopropan-2-yl]amino]-2-oxoethyl]amino]-3-[(2r)-2,3-di(hexadecanoyloxy)propyl]sulfanyl-1-oxopropan-2-yl Chemical compound O=C1C(SCCC(=O)NCCCOCCOCCOCCCNC(=O)COCC(=O)N[C@@H](CSC[C@@H](COC(=O)CCCCCCCCCCCCCCC)OC(=O)CCCCCCCCCCCCCCC)C(=O)NCC(=O)N[C@H](CO)C(=O)NCCCOCCOCCOCCCNC(=O)COCC(N)=O)CC(=O)N1CCNC(=O)CCCCCN\1C2=CC=C(S([O-])(=O)=O)C=C2CC/1=C/C=C/C=C/C1=[N+](CC)C2=CC=C(S([O-])(=O)=O)C=C2C1 UNILWMWFPHPYOR-KXEYIPSPSA-M 0.000 description 3
- YYROPELSRYBVMQ-UHFFFAOYSA-N 4-toluenesulfonyl chloride Chemical compound CC1=CC=C(S(Cl)(=O)=O)C=C1 YYROPELSRYBVMQ-UHFFFAOYSA-N 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- KEAYESYHFKHZAL-UHFFFAOYSA-N Sodium Chemical compound [Na] KEAYESYHFKHZAL-UHFFFAOYSA-N 0.000 description 3
- 238000007792 addition Methods 0.000 description 3
- KGNDCEVUMONOKF-UGPLYTSKSA-N benzyl n-[(2r)-1-[(2s,4r)-2-[[(2s)-6-amino-1-(1,3-benzoxazol-2-yl)-1,1-dihydroxyhexan-2-yl]carbamoyl]-4-[(4-methylphenyl)methoxy]pyrrolidin-1-yl]-1-oxo-4-phenylbutan-2-yl]carbamate Chemical compound C1=CC(C)=CC=C1CO[C@H]1CN(C(=O)[C@@H](CCC=2C=CC=CC=2)NC(=O)OCC=2C=CC=CC=2)[C@H](C(=O)N[C@@H](CCCCN)C(O)(O)C=2OC3=CC=CC=C3N=2)C1 KGNDCEVUMONOKF-UGPLYTSKSA-N 0.000 description 3
- WERYXYBDKMZEQL-UHFFFAOYSA-N butane-1,4-diol Chemical compound OCCCCO WERYXYBDKMZEQL-UHFFFAOYSA-N 0.000 description 3
- 125000004106 butoxy group Chemical group [*]OC([H])([H])C([H])([H])C(C([H])([H])[H])([H])[H] 0.000 description 3
- 229940126208 compound 22 Drugs 0.000 description 3
- 229940125833 compound 23 Drugs 0.000 description 3
- 229940125898 compound 5 Drugs 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 125000001301 ethoxy group Chemical group [H]C([H])([H])C([H])([H])O* 0.000 description 3
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- 235000017557 sodium bicarbonate Nutrition 0.000 description 3
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 3
- 239000012312 sodium hydride Substances 0.000 description 3
- 229910000104 sodium hydride Inorganic materials 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- UAOUIVVJBYDFKD-XKCDOFEDSA-N (1R,9R,10S,11R,12R,15S,18S,21R)-10,11,21-trihydroxy-8,8-dimethyl-14-methylidene-4-(prop-2-enylamino)-20-oxa-5-thia-3-azahexacyclo[9.7.2.112,15.01,9.02,6.012,18]henicosa-2(6),3-dien-13-one Chemical compound C([C@@H]1[C@@H](O)[C@@]23C(C1=C)=O)C[C@H]2[C@]12C(N=C(NCC=C)S4)=C4CC(C)(C)[C@H]1[C@H](O)[C@]3(O)OC2 UAOUIVVJBYDFKD-XKCDOFEDSA-N 0.000 description 2
- AOSZTAHDEDLTLQ-AZKQZHLXSA-N (1S,2S,4R,8S,9S,11S,12R,13S,19S)-6-[(3-chlorophenyl)methyl]-12,19-difluoro-11-hydroxy-8-(2-hydroxyacetyl)-9,13-dimethyl-6-azapentacyclo[10.8.0.02,9.04,8.013,18]icosa-14,17-dien-16-one Chemical compound C([C@@H]1C[C@H]2[C@H]3[C@]([C@]4(C=CC(=O)C=C4[C@@H](F)C3)C)(F)[C@@H](O)C[C@@]2([C@@]1(C1)C(=O)CO)C)N1CC1=CC=CC(Cl)=C1 AOSZTAHDEDLTLQ-AZKQZHLXSA-N 0.000 description 2
- GLGNXYJARSMNGJ-VKTIVEEGSA-N (1s,2s,3r,4r)-3-[[5-chloro-2-[(1-ethyl-6-methoxy-2-oxo-4,5-dihydro-3h-1-benzazepin-7-yl)amino]pyrimidin-4-yl]amino]bicyclo[2.2.1]hept-5-ene-2-carboxamide Chemical compound CCN1C(=O)CCCC2=C(OC)C(NC=3N=C(C(=CN=3)Cl)N[C@H]3[C@H]([C@@]4([H])C[C@@]3(C=C4)[H])C(N)=O)=CC=C21 GLGNXYJARSMNGJ-VKTIVEEGSA-N 0.000 description 2
- SZUVGFMDDVSKSI-WIFOCOSTSA-N (1s,2s,3s,5r)-1-(carboxymethyl)-3,5-bis[(4-phenoxyphenyl)methyl-propylcarbamoyl]cyclopentane-1,2-dicarboxylic acid Chemical compound O=C([C@@H]1[C@@H]([C@](CC(O)=O)([C@H](C(=O)N(CCC)CC=2C=CC(OC=3C=CC=CC=3)=CC=2)C1)C(O)=O)C(O)=O)N(CCC)CC(C=C1)=CC=C1OC1=CC=CC=C1 SZUVGFMDDVSKSI-WIFOCOSTSA-N 0.000 description 2
- QFLWZFQWSBQYPS-AWRAUJHKSA-N (3S)-3-[[(2S)-2-[[(2S)-2-[5-[(3aS,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]-3-methylbutanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-4-[1-bis(4-chlorophenoxy)phosphorylbutylamino]-4-oxobutanoic acid Chemical compound CCCC(NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)[C@@H](NC(=O)CCCCC1SC[C@@H]2NC(=O)N[C@H]12)C(C)C)P(=O)(Oc1ccc(Cl)cc1)Oc1ccc(Cl)cc1 QFLWZFQWSBQYPS-AWRAUJHKSA-N 0.000 description 2
- ONBQEOIKXPHGMB-VBSBHUPXSA-N 1-[2-[(2s,3r,4s,5r)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]oxy-4,6-dihydroxyphenyl]-3-(4-hydroxyphenyl)propan-1-one Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=CC(O)=C1C(=O)CCC1=CC=C(O)C=C1 ONBQEOIKXPHGMB-VBSBHUPXSA-N 0.000 description 2
- YSUIQYOGTINQIN-UZFYAQMZSA-N 2-amino-9-[(1S,6R,8R,9S,10R,15R,17R,18R)-8-(6-aminopurin-9-yl)-9,18-difluoro-3,12-dihydroxy-3,12-bis(sulfanylidene)-2,4,7,11,13,16-hexaoxa-3lambda5,12lambda5-diphosphatricyclo[13.2.1.06,10]octadecan-17-yl]-1H-purin-6-one Chemical compound NC1=NC2=C(N=CN2[C@@H]2O[C@@H]3COP(S)(=O)O[C@@H]4[C@@H](COP(S)(=O)O[C@@H]2[C@@H]3F)O[C@H]([C@H]4F)N2C=NC3=C2N=CN=C3N)C(=O)N1 YSUIQYOGTINQIN-UZFYAQMZSA-N 0.000 description 2
- TVTJUIAKQFIXCE-HUKYDQBMSA-N 2-amino-9-[(2R,3S,4S,5R)-4-fluoro-3-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-7-prop-2-ynyl-1H-purine-6,8-dione Chemical compound NC=1NC(C=2N(C(N(C=2N=1)[C@@H]1O[C@@H]([C@H]([C@H]1O)F)CO)=O)CC#C)=O TVTJUIAKQFIXCE-HUKYDQBMSA-N 0.000 description 2
- BPCQABSCRRCHFM-UHFFFAOYSA-N 2-tri(propan-2-yl)silyloxyethanol Chemical compound CC(C)[Si](C(C)C)(C(C)C)OCCO BPCQABSCRRCHFM-UHFFFAOYSA-N 0.000 description 2
- QBWKPGNFQQJGFY-QLFBSQMISA-N 3-[(1r)-1-[(2r,6s)-2,6-dimethylmorpholin-4-yl]ethyl]-n-[6-methyl-3-(1h-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-yl]-1,2-thiazol-5-amine Chemical compound N1([C@H](C)C2=NSC(NC=3C4=NC=C(N4C=C(C)N=3)C3=CNN=C3)=C2)C[C@H](C)O[C@H](C)C1 QBWKPGNFQQJGFY-QLFBSQMISA-N 0.000 description 2
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 2
- IGHBXJSNZCFXNK-UHFFFAOYSA-N 4-chloro-7-nitrobenzofurazan Chemical compound [O-][N+](=O)C1=CC=C(Cl)C2=NON=C12 IGHBXJSNZCFXNK-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 229940126657 Compound 17 Drugs 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 2
- LJOOWESTVASNOG-UFJKPHDISA-N [(1s,3r,4ar,7s,8s,8as)-3-hydroxy-8-[2-[(4r)-4-hydroxy-6-oxooxan-2-yl]ethyl]-7-methyl-1,2,3,4,4a,7,8,8a-octahydronaphthalen-1-yl] (2s)-2-methylbutanoate Chemical compound C([C@H]1[C@@H](C)C=C[C@H]2C[C@@H](O)C[C@@H]([C@H]12)OC(=O)[C@@H](C)CC)CC1C[C@@H](O)CC(=O)O1 LJOOWESTVASNOG-UFJKPHDISA-N 0.000 description 2
- LNUFLCYMSVYYNW-ZPJMAFJPSA-N [(2r,3r,4s,5r,6r)-2-[(2r,3r,4s,5r,6r)-6-[(2r,3r,4s,5r,6r)-6-[(2r,3r,4s,5r,6r)-6-[[(3s,5s,8r,9s,10s,13r,14s,17r)-10,13-dimethyl-17-[(2r)-6-methylheptan-2-yl]-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-3-yl]oxy]-4,5-disulfo Chemical compound O([C@@H]1[C@@H](COS(O)(=O)=O)O[C@@H]([C@@H]([C@H]1OS(O)(=O)=O)OS(O)(=O)=O)O[C@@H]1[C@@H](COS(O)(=O)=O)O[C@@H]([C@@H]([C@H]1OS(O)(=O)=O)OS(O)(=O)=O)O[C@@H]1[C@@H](COS(O)(=O)=O)O[C@H]([C@@H]([C@H]1OS(O)(=O)=O)OS(O)(=O)=O)O[C@@H]1C[C@@H]2CC[C@H]3[C@@H]4CC[C@@H]([C@]4(CC[C@@H]3[C@@]2(C)CC1)C)[C@H](C)CCCC(C)C)[C@H]1O[C@H](COS(O)(=O)=O)[C@@H](OS(O)(=O)=O)[C@H](OS(O)(=O)=O)[C@H]1OS(O)(=O)=O LNUFLCYMSVYYNW-ZPJMAFJPSA-N 0.000 description 2
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Images
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- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Nitrogen And Oxygen As The Only Ring Hetero Atoms (AREA)
- Plural Heterocyclic Compounds (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Description
本発明は、生体外、生体内におけるビスフェノールＡの挙動を検出可能な新規化合物及び該化合物を用いたビスフェノールＡの挙動の検出方法に関する。 The present invention relates to a novel compound capable of detecting the behavior of bisphenol A in vitro and in vivo, and a method for detecting the behavior of bisphenol A using the compound.
PCBやDDTなど環境ホルモンの多くは、当初、化学的な安定性やヒトへの急性毒性の低さから非常に優れた化合物と考えられ広く使用されていたが現在は使用禁止となっている。使用禁止に至るまでには、これらの化合物の定量法の確立や受容体の同定など、世界中で膨大な研究の蓄積が必要であった。ビスフェノールＡ（BpA）は実験的に環境ホルモン様の作用があると最初に証明された最も有名な物質で、歯科治療剤からプラスチックに至るまで広く利用されていた。現在のところ、ビスフェノールＡの代替品が開発されていないことと顕著な毒性が報告されていないことから、そのまま使用されているが、ビスフェノール類の環境中濃度の測定やその生理作用の検索は今後も盛んに行われると考えられる。前述のように、ビスフェノール類は、環境ホルモン様物質であることから、組織や細胞へ移行したり、受容体と結合したりする。したがって、ビスフェノール類の作用を特定するためには、環境中、細胞内、組織内、生体内でのビスフェノール類の挙動を追跡することが重要となる。 Many of the environmental hormones such as PCB and DDT were initially considered to be very excellent compounds due to their chemical stability and low acute toxicity to humans, but are now banned. Before the use was banned, it was necessary to accumulate enormous research all over the world, such as establishment of quantitative methods for these compounds and identification of receptors. Bisphenol A (BpA) is the most famous substance that was first experimentally proven to have an environmental hormone-like action, and was widely used from dental treatments to plastics. Currently, no substitute for bisphenol A has been developed and no significant toxicity has been reported, so it has been used as is. However, measurement of the environmental concentration of bisphenols and the search for their physiological effects will continue in the future. It is thought that it is also actively performed. As described above, since bisphenols are environmental hormone-like substances, they migrate to tissues and cells or bind to receptors. Therefore, in order to specify the action of bisphenols, it is important to track the behavior of bisphenols in the environment, in cells, in tissues, and in vivo.
したがって、ビスフェノール類を追跡、測定する方法は広く研究され、炭素同位体により標識されたビスフェノールＡを利用する方法（特許文献１参照）、ビスフェノールＡ に特異的に反応するモノクローナル抗体を利用する方法（特許文献２参照）などが報告されている。しかし、これまでの方法ではビスフェノールＡの検出が容易ではなかったため、より簡便な方法でビスフェノールＡを検出、測定する方法が望まれていた。
したがって、本発明は、ビスフェノールＡを簡便に検出する方法、そのための試薬を提供するものである。 Therefore, the present invention provides a method for easily detecting bisphenol A and a reagent therefor.
本発明者は、上記従来技術の問題点に鑑み鋭意検討を重ねた結果、ビスフェノールＡの水酸基に特定の蛍光標識基を結合した化合物を合成し、その挙動を確認したところ、この化合物がビスフェノールＡ様の挙動を示すことを見出し、本発明を完成させた。 As a result of intensive studies in view of the above-mentioned problems of the prior art, the present inventors synthesized a compound in which a specific fluorescent labeling group is bonded to the hydroxyl group of bisphenol A, and confirmed its behavior. As a result, the present invention has been completed.
すなわち、本発明は、下記の化合物、試薬及び方法を提供するものである。
項１．
下記式(1)
That is, the present invention provides the following compounds, reagents and methods.
The following formula (1)
又は下記式(3)で表される基
Or a group represented by the following formula (3)
を表す。Ｒ4は−ＯＨ基、上記式(2)で表される基、上記式(3)で表される基又は下記式(4)で表される基
Represents. R 4 represents an —OH group, a group represented by the above formula (2), a group represented by the above formula (3), or a group represented by the following formula (4).
を表す。〕
で表される化合物。
項２．
Ｒ1及びＲ2がともにメチル基を表す、項１に記載の化合物。
項３．
Ｒ4が−ＯＨ基、上記式(2)で表される基、上記式(3)で表される基又は−OCH2OCH3で表される基である項１又は２に記載の化合物。
項４．
項１〜３のいずれかに記載の化合物を含有するビスフェノールＡの挙動を検出するための試薬。
項５．
項１〜３のいずれかに記載の化合物の蛍光を検出する工程、これら化合物と結合したタンパク質の蛍光を検出する工程及びこれら化合物と結合したタンパク質の分子量を測定する工程からなる群から選択される少なくとも１種の工程を包含する、ビスフェノールＡの挙動を検出する方法。
Represents. ]
A compound represented by
Item 3. The compound according to
Item 4.
A reagent for detecting the behavior of bisphenol A containing the compound according to any one of
Item 4 is selected from the group consisting of a step of detecting fluorescence of the compound according to any one of 1 to 3, a step of detecting fluorescence of a protein bound to these compounds, and a step of measuring the molecular weight of the protein bound to these compounds A method for detecting the behavior of bisphenol A comprising at least one step.
本発明の化合物は新規化合物であり、上記式(1)に表されるように、ビスフェノール構造の一方の水酸基に、アルキレン基を介して、蛍光標識基（ニトロベンゾ-2-オキサ-1,3-ジアゾール基（NBD基））又は光親和性標識基（4-トリフルオロメチルジアジリニルフェノキシ基が結合し、もう一方が水酸基であるか、又は水酸基にアルキレン基を介して蛍光標識基（ニトロベンゾ-2-オキサ-1,3-ジアゾール基（NBD基））、光親和性標識基（4-トリフルオロメチルジアジリニルフェノキシ基）もしくはアルコキシ基が結合した構造を有する。 The compound of the present invention is a novel compound, and, as represented by the above formula (1), a fluorescent labeling group (nitrobenzo-2-oxa-1,3- Diazole group (NBD group)) or photoaffinity labeling group (4-trifluoromethyldiazilinylphenoxy group is bonded, and the other is a hydroxyl group, or a fluorescent labeling group (nitrobenzo- It has a structure in which a 2-oxa-1,3-diazole group (NBD group)), a photoaffinity labeling group (4-trifluoromethyldiazilinylphenoxy group) or an alkoxy group is bonded.
蛍光標識基はある特定の光を吸収することで活性化状態へ励起され、これが基底状態へ戻る際に蛍光を発するため、この蛍光を検出することによって本発明の化合物の挙動を確認することが可能である。 A fluorescent labeling group is excited to an activated state by absorbing certain light, and emits fluorescence when it returns to the ground state. Therefore, the behavior of the compound of the present invention can be confirmed by detecting this fluorescence. Is possible.
また、光親和性標識基は光親和性を有する。光親和性標識基は、光照射（360nm）によってジアジリン環が開環してカルベンが生成する。これが容易に、受容体や酵素などのビスフェノールＡを捕捉する物質に存在する官能基と結合し、受容体や酵素などとの付加物を生成する。このようにして生成した光親和性標識基に基づく受容体や酵素などとの付加物は、クロマトグラフィーや電気泳動で分離し、質量分析装置等で解析することが可能であり、本発明の化合物の挙動を確認することが可能である。したがって、上記式(3)で表される基を有する本発明の化合物は、光照射によって受容体や酵素を捕捉することができる。 The photoaffinity labeling group has photoaffinity. In the photoaffinity labeling group, the diazirine ring is opened by light irradiation (360 nm) to generate carbene. This easily binds to a functional group present in a substance that captures bisphenol A, such as a receptor or an enzyme, and generates an adduct with the receptor or the enzyme. The adduct with a receptor or enzyme or the like based on the photoaffinity labeling group thus generated can be separated by chromatography or electrophoresis and analyzed by a mass spectrometer or the like, and the compound of the present invention It is possible to confirm the behavior of Therefore, the compound of the present invention having a group represented by the above formula (3) can capture a receptor and an enzyme by light irradiation.
さらに、本発明の化合物はビスフェノールＡと同様な挙動を示す特性を備えている。このため、本発明の化合物の挙動を検出することによって、ビスフェノールＡの挙動、例えば、ビスフェノールＡの結合部位、相互作用などを確認することが可能である。したがって、本発明の化合物は、ビスフェノールＡの挙動を検出する試薬として非常に有用である。 Furthermore, the compound of this invention is equipped with the characteristic which shows the same behavior as bisphenol A. Therefore, by detecting the behavior of the compound of the present invention, it is possible to confirm the behavior of bisphenol A, for example, the binding site of bisphenol A, the interaction, and the like. Therefore, the compound of the present invention is very useful as a reagent for detecting the behavior of bisphenol A.
本発明の化合物において、Ｒ1は水素原子、メチル基又はエチル基を表し、好ましくはメチル基である。さらに、本発明の化合物において、Ｒ2は水素原子、メチル基又はエチル基を表し、好ましくはメチル基である。 In the compound of the present invention, R 1 represents a hydrogen atom, a methyl group or an ethyl group, preferably a methyl group. Furthermore, in the compound of the present invention, R 2 represents a hydrogen atom, a methyl group or an ethyl group, preferably a methyl group.
本発明の化合物においてＲ3は上記式(2)又は(3)で表される基である。また、本発明の化合物において、ｎは２〜４の整数である。すなわち、ビスフェノール構造と蛍光標識基との間に介在するアルキレン基は炭素数２〜４である。また、本発明の化合物において、ｍは２〜４の整数である。すなわち、ビスフェノール構造と光親和性標識基との間に介在するアルキレン基は炭素数２〜４である。好ましくは、ｎ及びｍがともに２である。 In the compound of the present invention, R 3 is a group represented by the above formula (2) or (3). Moreover, in the compound of this invention, n is an integer of 2-4. That is, the alkylene group interposed between the bisphenol structure and the fluorescent labeling group has 2 to 4 carbon atoms. Moreover, in the compound of this invention, m is an integer of 2-4. That is, the alkylene group interposed between the bisphenol structure and the photoaffinity labeling group has 2 to 4 carbon atoms. Preferably, n and m are both 2.
本発明の化合物においてＲ4は−ＯＨ基、上記式(2)で表される基、上記式(3)で表される基又は上記式(4)で表される基である。上記式(4)において、ｌは１〜３の整数であり、Ｒ5は炭素数１〜３のアルキル基、好ましくはメチル基である。上記式(4)においてはｌが１、Ｒ5がメチル基であること、すなわち、−ＯＣＨ2−ＯＣＨ3が好ましい。好ましい本発明の化合物の具体的な例としては、下記式(5)〜(10)で表される化合物が挙げられる。 In the compound of the present invention, R 4 is an —OH group, a group represented by the above formula (2), a group represented by the above formula (3), or a group represented by the above formula (4). In the above formula (4), l is an integer of 1 to 3, and R 5 is an alkyl group having 1 to 3 carbon atoms, preferably a methyl group. In the above formula (4), l is 1 and R 5 is a methyl group, that is, —OCH 2 —OCH 3 is preferable. Specific examples of preferable compounds of the present invention include compounds represented by the following formulas (5) to (10).
ビスフェノールＡに存在する２つのフェノール性水酸基のうちの一方を保護した保護体において、残るフェノール性の水酸基にエーテル結合を構築しながら光親和性標識基を導入する。また、同様にしてアルカンジオールを用いてエーテル結合を構築した後、残るアルコールをアミノ基に変換し、その後、蛍光標識基を導入する。さらに、フェノール性水酸基の保護基を除去した後、光親和性標識基を導入する。 In the protected body in which one of the two phenolic hydroxyl groups present in bisphenol A is protected, a photoaffinity labeling group is introduced while an ether bond is constructed on the remaining phenolic hydroxyl group. Similarly, after an ether bond is constructed using alkanediol, the remaining alcohol is converted into an amino group, and then a fluorescent labeling group is introduced. Furthermore, after removing the protecting group for the phenolic hydroxyl group, a photoaffinity labeling group is introduced.
本発明の試薬は、本発明の化合物を必須成分として含有するものである。本発明の試薬は、本発明の化合物の検出を妨げない範囲で使用目的に応じた他の成分を適当な量で適宜含むことができる。他の成分の一例としては、ジメチルスルホキシド、ジメチルホルムアミド、ジオキサンなどの有機溶媒が挙げられる。本発明の試薬は必要に応じて上記有機溶媒やその他の担体を含有し、細胞、組織、器官、生体、環境水などの検出対象物に適当な方法で添加され、その後、本発明化合物の発する蛍光又は光照射によって生ずる本発明化合物とこれに隣接する物質とが結合した物質の分子量を測定することによって、ビスフェノールＡの挙動を予測することが可能である。 The reagent of the present invention contains the compound of the present invention as an essential component. The reagent of the present invention can appropriately contain other components in an appropriate amount depending on the purpose of use within the range that does not interfere with the detection of the compound of the present invention. Examples of other components include organic solvents such as dimethyl sulfoxide, dimethylformamide, and dioxane. The reagent of the present invention contains the above organic solvent and other carriers as necessary, and is added to a detection target such as cells, tissues, organs, living bodies, and environmental water by an appropriate method, and then the compound of the present invention is emitted. The behavior of bisphenol A can be predicted by measuring the molecular weight of a substance in which the compound of the present invention produced by fluorescence or light irradiation and a substance adjacent thereto are bound.
本発明のビスフェノールＡの挙動を検出する方法は、本発明の化合物に必要に応じて上記有機溶媒やその他の担体を組み合わせ、細胞、組織、器官、生体、環境水などの検出対象物に適当な方法で添加され、その後、本発明化合物の発する蛍光、又は光照射によって活性化された本発明化合物と結合した物質の分子量を測定することによって、本発明化合物の挙動を検出し、この挙動をビスフェノールＡの挙動として特定する。 The method for detecting the behavior of bisphenol A of the present invention is suitable for a detection target such as a cell, tissue, organ, living body, and environmental water by combining the compound of the present invention with the above organic solvent and other carriers as necessary. The behavior of the compound of the present invention is detected by measuring the molecular weight of the substance added to the compound of the present invention and subsequently bound by the fluorescence emitted by the compound of the present invention or activated by light irradiation. It is specified as the behavior of A.
本発明の化合物は、蛍光標識基を有するため、細胞等の生体内又は環境水等の生体外でも蛍光を検出することによって、その挙動を検出可能であり、また、光親和性標識基を有するため、細胞等の生体内又は環境水等の生体外でも光照射によって、その挙動を検出可能である。また、本発明の化合物はビスフェノールＡと同じ挙動を示すため、本発明の化合物の挙動を把握することによって、ビスフェノールＡの挙動も把握することが可能となる。 Since the compound of the present invention has a fluorescent labeling group, its behavior can be detected by detecting fluorescence in vivo such as cells or in vitro such as environmental water, and also has a photoaffinity labeling group. Therefore, the behavior can be detected by irradiation with light in a living body such as a cell or outside a living body such as environmental water. Moreover, since the compound of this invention shows the same behavior as bisphenol A, it becomes possible to grasp | ascertain the behavior of bisphenol A by grasping | ascertaining the behavior of the compound of this invention.
以下、実施例により本発明をより詳細に説明するが、本発明はこれらに限定されるものではない。 EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, this invention is not limited to these.
合成には下記の装置及び試薬を使用した。
＜分析装置＞
NMR：α−400（日本電子社製）
IR：FT/IR-5300（日本分光社製）
MASS：JMS-T100LC（日本電子社製）
＜クロマトグラフィー＞
TLC：Art5717Merck Kieselgel 60F254
分取用：Art5717Merck Kieselgel 60F254
シリカゲルクロマトグラフィー：BW-200（富士デビィソン社製）
The following equipment and reagents were used for the synthesis.
<Analyzer>
NMR: α-400 (manufactured by JEOL Ltd.)
IR: FT / IR-5300 (manufactured by JASCO)
MASS: JMS-T100LC (manufactured by JEOL Ltd.)
<Chromatography>
TLC: Art5717Merck Kieselgel 60F254
For fractionation: Art5717Merck Kieselgel 60F254
Silica gel chromatography: BW-200 (Fuji Devison)
実施例１
本発明の化合物（11）、（18）、（20）、（22）を図１に示す合成スキームにしたがい製造した。
Example 1
Compounds (11), (18), (20) and (22) of the present invention were produced according to the synthetic scheme shown in FIG.
2-(p-hydroxyphenyl)-2-(p-(methylenedioxymethyl)phenyl)propane (2)の製造
アルゴン雰囲気下、ビスフェノールＡ(1)(100 g, 438 mmol) のDMF溶液(2.0 l)を0 ℃に冷却した後、水素化ナトリウム（油性）(21.0 g, 526 mmol)を加え、室温で１時間攪拌した後、メトキシメチルクロリド(40 ml, 526 mmol)を滴下し、室温で１５分間攪拌した。反応混合物を0 ℃に冷却した後、蒸留水を滴下し、ジエチルエーテルで抽出した。有機層を飽和食塩水で洗浄後、硫酸マグネシウムで乾燥させ、減圧濃縮した。残留物をシリカゲルカラムクロマトグラフィー(ヘキサン中3%から50%の酢酸エチル)で精製し、化合物2 (50.7 g, 収率43%)を得た。
1H NMR (CDCl3, 400 MHz) d 7.13 (AB-q, J = 8.5 Hz, 2H), 7.09 (AB-q, J = 8.3 Hz, 2H), 7.03 (AB-q, J = 8.5 Hz, 2H), 6.72 (AB-q, J = 8.3 Hz, 2H), 5.15 (s, 2H) , 3.50 (s, 3H), 1.63 (s, 6H);
13C NMR (CDCl3, 100 MHz) d 162.8, 153.8, 127.8, 127.7, 115.6, 114.7, 94.5, 55.9, 41.7, 31.0;
IR (NaCl neat, cm-1) 3387, 2967, 1611, 1510, 1235, 1179, 1152, 1013, 833;
ESI HRMS m/z C17H20O3 [M-H]- 271.1334(計算値), 271.1326(測定値)。
Preparation of 2- (p-hydroxyphenyl) -2- (p- (methylenedioxymethyl) phenyl) propane (2) DMF solution (2.0 l) of bisphenol A (1) (100 g, 438 mmol) at 0 ℃ under argon atmosphere After cooling to 0, sodium hydride (oil) (21.0 g, 526 mmol) was added and stirred at room temperature for 1 hour, methoxymethyl chloride (40 ml, 526 mmol) was added dropwise, and the mixture was stirred at room temperature for 15 minutes. After the reaction mixture was cooled to 0 ° C., distilled water was added dropwise and extracted with diethyl ether. The organic layer was washed with saturated brine, dried over magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (3% to 50% ethyl acetate in hexane) to give compound 2 (50.7 g, 43% yield).
1 H NMR (CDCl 3 , 400 MHz) d 7.13 (AB-q, J = 8.5 Hz, 2H), 7.09 (AB-q, J = 8.3 Hz, 2H), 7.03 (AB-q, J = 8.5 Hz, 2H), 6.72 (AB-q, J = 8.3 Hz, 2H), 5.15 (s, 2H), 3.50 (s, 3H), 1.63 (s, 6H);
13 C NMR (CDCl 3 , 100 MHz) d 162.8, 153.8, 127.8, 127.7, 115.6, 114.7, 94.5, 55.9, 41.7, 31.0;
IR (NaCl neat, cm -1 ) 3387, 2967, 1611, 1510, 1235, 1179, 1152, 1013, 833;
ESI HRMS m / z C 17 H 20 O 3 [MH] - 271.1334 ( calcd), 271.1326 (measured value).
2-(triisopropylsilyloxy)ethanol (5)の製造
アルゴン雰囲気下、エチレングリコール(3) (5.00 g, 80.5 mmol)のDMF(400 ml)溶液を0 ℃に冷却した後、イミダゾール (6.60 g, 96.7 mmol)を加えた。0 ℃で15分間攪拌し、トリイソプロピルシリルクロリド(20.5 ml, 96.7 mmol)を滴下した。反応混合物を室温で2分間攪拌した後、0 ℃に冷却し、飽和炭酸水素ナトリウム水溶液を加え、ジエチルエーテルで抽出した。有機層を飽和食塩水で洗浄後、硫酸マグネシウムで乾燥させ、減圧濃縮した。残留物をシリカゲルクロマトグラフィー(ヘキサン中1%から33%の酢酸エチル)により精製し、化合物5(16.4 g, 収率88%)を得た。
1H NMR (CDCl3, 400 MHz) d 3.80 (t, J = 4.4 Hz, 2H), 3.64-3.68 (m, 2H), 1.05-1.67 (m, 21H);
13C NMR (CDCl3, 100 MHz) d 64.2, 63.8, 17.9, 11.9;
IR (NaCl neat, cm-1) 3374, 2944, 2868, 1464, 1385, 789;
ESI HRMS m/z C11H26O2Si [M + Na]+ 241.1600(計算値), 24.1599(測定値)
Preparation of 2- (triisopropylsilyloxy) ethanol (5) Under a argon atmosphere, a solution of ethylene glycol (3) (5.00 g, 80.5 mmol) in DMF (400 ml) was cooled to 0 ° C, and then imidazole (6.60 g, 96.7 mmol) Was added. The mixture was stirred at 0 ° C. for 15 minutes, and triisopropylsilyl chloride (20.5 ml, 96.7 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 2 minutes, cooled to 0 ° C., saturated aqueous sodium hydrogen carbonate solution was added, and the mixture was extracted with diethyl ether. The organic layer was washed with saturated brine, dried over magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (1% to 33% ethyl acetate in hexanes) to give compound 5 (16.4 g, 88% yield).
1 H NMR (CDCl 3 , 400 MHz) d 3.80 (t, J = 4.4 Hz, 2H), 3.64-3.68 (m, 2H), 1.05-1.67 (m, 21H);
13 C NMR (CDCl 3 , 100 MHz) d 64.2, 63.8, 17.9, 11.9;
IR (NaCl neat, cm -1 ) 3374, 2944, 2868, 1464, 1385, 789;
ESI HRMS m / z C 11 H 26 O 2 Si [M + Na] + 241.1600 (calculated value), 24.1599 (measured value)
2-(ethylenedioxyisopropylsilyoxy-p-phenyl)-2-(p-(methylenedioxymethyl)phenyl)propane (7)の製造
アルゴン雰囲気下、化合物5（100 mg, 0.431 mmol）のTHF(2.0 ml)溶液を0 ℃に冷却した後、トリフェニルホスフィン(170 mg, 0.647 mmol)を加え、ジイソプロピルアザジカルボキシレート(0.14 ml, 0.686 mmol)を滴下し、0 ℃で40分攪拌した後、フェノールを滴下した。反応混合物を室温で3時間攪拌した後、減圧濃縮し、化合物7を得た。精製は行わず次の反応に用いた。
Preparation of 2- (ethylenedioxyisopropylsilyoxy-p-phenyl) -2- (p- (methylenedioxymethyl) phenyl) propane (7) In an argon atmosphere, a THF (2.0 ml) solution of compound 5 (100 mg, 0.431 mmol) was brought to 0 ° C. After cooling, triphenylphosphine (170 mg, 0.647 mmol) was added, diisopropyl azadicarboxylate (0.14 ml, 0.686 mmol) was added dropwise, and the mixture was stirred at 0 ° C. for 40 minutes, and then phenol was added dropwise. The reaction mixture was stirred at room temperature for 3 hours and then concentrated under reduced pressure to give
2-(p-(2'-(p-(methylenedioxymethyl)phenyl)propane-2'-yl)phenoxy)ethanol (9)の製造
アルゴン雰囲気下、化合物7のTHF(5.0 ml)溶液にテトラブチルアンモニウムフロオリド(360 mg, 1.38 mmol)を加えた。反応混合物を室温で３時間攪拌した後、蒸留水を加え、ジエチルエーテルで抽出した。有機層を飽和食塩水で洗浄後、硫酸マグネシウムで乾燥させ、減圧濃縮した。残留物をシリカゲルカラムクロマトグラフィー(ヘキサン中13%から17%の酢酸エチル)で精製し、化合物9 (97 mg, 収率70%)を得た。
1H NMR (CDCl3, 400 MHz) d 7.15 (AB-q, J = 8.8 Hz, 2H), 7.13 (AB-q, J = 9.0 Hz, 2H), 6.93 (AB-q, J = 9.0 Hz, 2H), 6.82 (AB-q, J = 9.0 Hz, 2H), 5.15 (s, 2H) , 4.06 (t, J = 4.1 Hz, 2H), 3.92-3.96 (m, 2H), 3.47 (s, 3H), 1.62 (s, 6H);
13C NMR (CDCl3, 100 MHz) d 155.4, 155.1, 144.3, 143.5, 127.8, 127.7,115.7, 113.9, 94.5, 69.1, 61.5, 56.0, 41.8, 31.0;
IR (NaCl neat, cm-1) 3426, 2965, 1609, 1510, 1238, 1179, 1154, 1011, 833;
ESI HRMS m/z C19H24O4 [M + Na]+ 339.1572(計算値), 339.1564(測定値)
2- (p- (2 '-(p- (methylenedioxymethyl) phenyl) propane-2'-yl) phenoxy) ethanol (9) Preparation of tetrabutylammonium fluoride in THF (5.0 ml) solution under argon atmosphere Orido (360 mg, 1.38 mmol) was added. The reaction mixture was stirred at room temperature for 3 hours, distilled water was added, and the mixture was extracted with diethyl ether. The organic layer was washed with saturated brine, dried over magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (13% to 17% ethyl acetate in hexane) to give compound 9 (97 mg, yield 70%).
1 H NMR (CDCl 3 , 400 MHz) d 7.15 (AB-q, J = 8.8 Hz, 2H), 7.13 (AB-q, J = 9.0 Hz, 2H), 6.93 (AB-q, J = 9.0 Hz, 2H), 6.82 (AB-q, J = 9.0 Hz, 2H), 5.15 (s, 2H), 4.06 (t, J = 4.1 Hz, 2H), 3.92-3.96 (m, 2H), 3.47 (s, 3H ), 1.62 (s, 6H);
13 C NMR (CDCl 3 , 100 MHz) d 155.4, 155.1, 144.3, 143.5, 127.8, 127.7, 115.7, 113.9, 94.5, 69.1, 61.5, 56.0, 41.8, 31.0;
IR (NaCl neat, cm -1 ) 3426, 2965, 1609, 1510, 1238, 1179, 1154, 1011, 833;
ESI HRMS m / z C 19 H 24 O 4 [M + Na] + 339.1572 (calculated value), 339.1564 (measured value)
3-(ethylenedioxy-(p-(2'-(p-(methylenedioxymethyl)phenyl)propane-2'-yl)phenyl)-3-trifluoromethyl-3H-diazirine (11)の製造
アルゴン雰囲気下、化合物9(230 mg, 0.730 mmol) のTHF溶液(4.0 ml)を0 ℃に冷却した後、トリフェニルホスフィン(286 mg, 1.09 mmol)を加え、ジイソプロピルアザジカルボキシレート(0.21 ml, 1.09 mmol)を滴下した。0 ℃で30分間攪拌した後、化合物13(220 mg, 1.09 mmol)を滴下した。反応混合物を室温で12時間攪拌した後、減圧濃縮した。残留物をシリカゲルクロマトグラフィー(ヘキサン中1%から5%の酢酸エチル)で精製し、化合物11(354 mg, 収率97%)を得た。
1H NMR (CDCl3, 400 MHz) d 7.15 (AB-q, J = 8.8 Hz, 2H), 7.14 (AB-q, J = 9.2 Hz, 2H), 7.13 (AB-q, J = 9.0 Hz, 2H),6.93 (AB-q, J = 9.0 Hz, 2H), 6.92 (AB-q, J = 9.0 Hz, 2H), 6.83 (AB-q, J = 9.0 Hz, 2H), 5.14 (s, 2H) , 4.28 (m, 4H), 3.46 (s, 3H), 1.63 (s, 6H);
13C NMR (CDCl3, 100 MHz) d 159.7, 156.3, 155.1, 144.3, 143.6, 130.5, 128.1, 127.8, 127.7, 122.2 (q, J C-F = 274.2 Hz), 121.3, 115.0, 114.6, 114.0, 94.5, 66.7, 66.2, 55.9, 41.7, 31.0;
IR (NaCl neat, cm-1) 2965, 1736, 1622, 1582, 1508, 1443, 1238, 1182, 1152, 833;
ESI HRMS m/z C27H27F3 N 2O4 [M + Na]+ 523.1821(計算値), 532.1835(測定値)
3- (ethylenedioxy- (p- (2 '-(p- (methylenedioxymethyl) phenyl) propane-2'-yl) phenyl) -3-trifluoromethyl-3H-diazirine (11) Preparation of Compound 9 (230 mg, 0.730 mmol) in THF (4.0 ml) was cooled to 0 ° C., triphenylphosphine (286 mg, 1.09 mmol) was added, and diisopropyl azadicarboxylate (0.21 ml, 1.09 mmol) was added dropwise. After stirring at 30 ° C. for 30 minutes, Compound 13 (220 mg, 1.09 mmol) was added dropwise, the reaction mixture was stirred at room temperature for 12 hours and then concentrated under reduced pressure, and the residue was chromatographed on silica gel (1% to 5% in hexane). Of ethyl acetate) to obtain Compound 11 (354 mg, 97% yield).
1 H NMR (CDCl 3 , 400 MHz) d 7.15 (AB-q, J = 8.8 Hz, 2H), 7.14 (AB-q, J = 9.2 Hz, 2H), 7.13 (AB-q, J = 9.0 Hz, 2H), 6.93 (AB-q, J = 9.0 Hz, 2H), 6.92 (AB-q, J = 9.0 Hz, 2H), 6.83 (AB-q, J = 9.0 Hz, 2H), 5.14 (s, 2H ), 4.28 (m, 4H), 3.46 (s, 3H), 1.63 (s, 6H);
13 C NMR (CDCl 3 , 100 MHz) d 159.7, 156.3, 155.1, 144.3, 143.6, 130.5, 128.1, 127.8, 127.7, 122.2 (q, J CF = 274.2 Hz), 121.3, 115.0, 114.6, 114.0, 94.5, 66.7, 66.2, 55.9, 41.7, 31.0;
IR (NaCl neat, cm -1 ) 2965, 1736, 1622, 1582, 1508, 1443, 1238, 1182, 1152, 833;
ESI HRMS m / z C 27 H 27 F 3 N 2 O 4 [M + Na] + 523.1821 (calculated value), 532.1835 (measured value)
2-(p-(2'-azidoethoxy)phenyl)-2-(p-(methylenedioxymethyl)phenyl)propane (14)の製造
アルゴン雰囲気下、化合物9(0.559 g, 1.77 mmol) のTHF (7.0 ml) 溶液を0 ℃に冷却した後、ここへトリフェニルホスフィン(1.02g, 3.89 mmol)を加え、ジイソプロピルアザジカルボキシレート(0.77 ml, 3.89 mmol)を滴下し、0 ℃で5分攪拌した後、ジフェニルホスホリルアジド(0.84 ml, 3.89 mmol)を滴下した。反応混合物を室温で3時間攪拌した後、減圧濃縮し化合物14を得た。精製は行わず次の反応に用いた。
Preparation of 2- (p- (2'-azidoethoxy) phenyl) -2- (p- (methylenedioxymethyl) phenyl) propane (14) Compound 9 (0.559 g, 1.77 mmol) in THF (7.0 ml) under an argon atmosphere Was cooled to 0 ° C., triphenylphosphine (1.02 g, 3.89 mmol) was added thereto, diisopropyl azadicarboxylate (0.77 ml, 3.89 mmol) was added dropwise thereto, and the mixture was stirred at 0 ° C. for 5 minutes. Azide (0.84 ml, 3.89 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 3 hours and then concentrated under reduced pressure to give
2-(p-(2'-aminoethoxy)phenyl)-2-(p-(methylenedioxymethyl)phenyl)propane (16)の製造
アルゴン雰囲気下、化合物14のTHF(16.0 ml)溶液にトリフェニルホスフィン(0.927 g, 3.53 mmol)を加えた。60 ℃に昇温した後、蒸留水(1.77 ml)を加え、60 ℃で3時間攪拌した。反応混合物を室温に冷却した後、蒸留水を加え、酢酸エチルで抽出した。有機層を飽和食塩水で洗浄後、硫酸マグネウシムで乾燥させ、減圧濃縮し化合物16を得た。精製は行わず次の反応に用いた。
Preparation of 2- (p- (2'-aminoethoxy) phenyl) -2- (p- (methylenedioxymethyl) phenyl) propane (16) Triphenylphosphine (0.927 g) in THF (16.0 ml) solution of
2-(p-(methylenedioxymethyl)phenyl)-2-(p-(2'-(N-(7''-nitrobenzo-2''-oxa-1'',3''-diazol-4''-yl)amino)ethoxy)phenylpropane (18)の製造
アルゴン雰囲気下、化合物16のTHF(5.0 ml)溶液に、トリエチルアミン(0.22 ml, 1.57 mmol)を滴下した。0 ℃に冷却した後、7-ニトロ-4-ベンゾフラザニルクロリド(0.25 g, 1.26 mmol)を加えた。反応混合物を室温で30分攪拌した後、飽和塩化アンモニウム水溶液を加え、ジエチルエーテルで抽出した。有機層を飽和食塩水で洗浄し、硫酸マグネシウムで乾燥した後、減圧濃縮した。残留物をシリカゲルクロマトグラフィー(ヘキサン中9%から25%の酢酸エチル)で精製し、化合物18(0.45 g, 収率52%)を得た。
1H NMR (CDCl3, 400 MHz) d 8.49 (d, J = 8.5 Hz, 1H), 7.17 (AB-q, J = 8.8 Hz, 2H), 7.12 (AB-q, J = 8.8 Hz, 2H), 6.93 (AB-q, J = 8.8 Hz, 2H), 6.83 (AB-q, J = 8.8 Hz, 2H), 6.61 (br-s, 1H), 6.29 (d, J = 8.5 Hz, 1H), 5.14 (s, 2H) , 4.30 (t, J = 4.8 Hz, 2H), 3.92 (dt, J = 4.9, 5.0 Hz, 2H), 3.47 (s, 3H), 1.63 (s, 6H);
13C NMR (CDCl3, 100 MHz) d 155.7, 155.1, 144.4, 144.3, 144.1, 143.8, 136.2, 128.0, 127.7, 115.7, 113.9, 99.0, 94.5, 65.2, 56.0, 43.3, 41.8, 31.9, 25.5;
IR (NaCl neat, cm-1) 2928, 1580, 1510, 1424, 1217, 1154, 1013;
ESI HRMS m/z C25H26N 4O6 [M − H]- 477.1774(計算値), 477.1768(測定値)
2- (p- (methylenedioxymethyl) phenyl) -2- (p- (2 '-(N- (7``-nitrobenzo-2''-oxa-1'',3''-diazol-4''- Preparation of (yl) amino) ethoxy) phenylpropane (18) Triethylamine (0.22 ml, 1.57 mmol) was added dropwise to a THF (5.0 ml) solution of
1 H NMR (CDCl 3 , 400 MHz) d 8.49 (d, J = 8.5 Hz, 1H), 7.17 (AB-q, J = 8.8 Hz, 2H), 7.12 (AB-q, J = 8.8 Hz, 2H) , 6.93 (AB-q, J = 8.8 Hz, 2H), 6.83 (AB-q, J = 8.8 Hz, 2H), 6.61 (br-s, 1H), 6.29 (d, J = 8.5 Hz, 1H), 5.14 (s, 2H), 4.30 (t, J = 4.8 Hz, 2H), 3.92 (dt, J = 4.9, 5.0 Hz, 2H), 3.47 (s, 3H), 1.63 (s, 6H);
13 C NMR (CDCl 3 , 100 MHz) d 155.7, 155.1, 144.4, 144.3, 144.1, 143.8, 136.2, 128.0, 127.7, 115.7, 113.9, 99.0, 94.5, 65.2, 56.0, 43.3, 41.8, 31.9, 25.5;
IR (NaCl neat, cm -1 ) 2928, 1580, 1510, 1424, 1217, 1154, 1013;
ESI HRMS m / z C 25 H 26 N 4 O 6 [M - H] - 477.1774 ( calcd), 477.1768 (measured value)
2-(p-hydroxyphenyl)-2-(p-(2'-(N-(7''-nitrobenzo-2''-oxa-1'',3''-diazol-4''-yl)amino)ethoxy)phenylpropane (20)の製造
化合物18(170 mg, 0.355 mmol) の酢酸 (0.70 ml) 溶液に、2規定塩酸水溶液(0.36 ml, 0.720 mmol)を加えた。反応混合物を室温で3日間攪拌し、蒸留水を加え、ジエチルエーテルで抽出した。有機層を飽和炭酸水素ナトリウム溶液で洗浄した後、飽和食塩水で洗浄し、硫酸マグネシウムで乾燥した後、減圧濃縮した。残留物をシリカゲルカラムクロマトグラフィー(ヘキサン中0%から9%の酢酸エチル)で精製し、化合物20(126 mg, 収率82%)を得た。
1H NMR (CDCl3, 400 MHz) d 8.51 (d, J = 8.5 Hz, 1H), 7.27 (AB-q, J =8.8 Hz, 2H), 7.07 (AB-q, J = 8.8 Hz, 2H), 6.83 (AB-q, J = 8.8 Hz, 2H), 6.73 (AB-q, J = 8.8 Hz, 2H), 6.54 (br-s, 1H), 6.28 (d, J = 8.5 Hz, 1H), 4.30 (t, J = 4.9 Hz, 2H), 3.90 (dt, J = 4.9, 5.0 Hz, 2H), 1.63 (s, 6H);
13C NMR (CDCl3, 100 MHz) d 155.7, 153.4, 144.6, 144.3, 143.8, 143.6, 143.0, 136.1, 128.0, 127.9, 114.7, 113.9, 99.9, 65.2, 43.3, 42.0, 41.8, 31.0;
IR (KBr disk, cm-1) 3337, 2965, 1620, 1584, 1508, 1443, 1306, 1184, 1136, 999, 833; ESI HRMS m/z C23H22 N 4O5 [M − H]- 433.1512(計算値), 433.1506(測定値)
2- (p-hydroxyphenyl) -2- (p- (2 '-(N- (7``-nitrobenzo-2''-oxa-1'',3''-diazol-4''-yl) amino ) ethoxy) phenylpropane (20) Preparation Compound 18 (170 mg, 0.355 mmol) in acetic acid (0.70 ml) was added with 2N aqueous hydrochloric acid (0.36 ml, 0.720 mmol), and the reaction mixture was stirred at room temperature for 3 days. Distilled water was added, and the mixture was extracted with diethyl ether.The organic layer was washed with saturated sodium hydrogen carbonate solution, washed with saturated brine, dried over magnesium sulfate, and concentrated under reduced pressure. Purification by chromatography (0% to 9% ethyl acetate in hexanes) provided compound 20 (126 mg, 82% yield).
1 H NMR (CDCl 3 , 400 MHz) d 8.51 (d, J = 8.5 Hz, 1H), 7.27 (AB-q, J = 8.8 Hz, 2H), 7.07 (AB-q, J = 8.8 Hz, 2H) , 6.83 (AB-q, J = 8.8 Hz, 2H), 6.73 (AB-q, J = 8.8 Hz, 2H), 6.54 (br-s, 1H), 6.28 (d, J = 8.5 Hz, 1H), 4.30 (t, J = 4.9 Hz, 2H), 3.90 (dt, J = 4.9, 5.0 Hz, 2H), 1.63 (s, 6H);
13 C NMR (CDCl 3 , 100 MHz) d 155.7, 153.4, 144.6, 144.3, 143.8, 143.6, 143.0, 136.1, 128.0, 127.9, 114.7, 113.9, 99.9, 65.2, 43.3, 42.0, 41.8, 31.0;
IR (KBr disk, cm -1 ) 3337, 2965, 1620, 1584, 1508, 1443, 1306, 1184, 1136, 999, 833; ESI HRMS m / z C 23 H 22 N 4 O 5 [M − H] - 433.1512 (calculated value), 433.1506 (measured value)
3-trifluoromethyl-3-(ethylenedioxytriisopropylsiloxy-p-phenyl)-3H-diazirine (24)の製造
アルゴン雰囲気下、化合物5(300 mg, 1.37 mmol) のTHF溶液(15 ml)を0 ℃に冷却した後、トリフェニルホスフィン(540 mg, 2.06 mmol)を加え、ジイソプロピルアザジカルボキシレート(0.41 ml, 2.06 mmol)を滴下した。反応混合物を0 ℃で10分間攪拌した後、化合物13(333 mg, 1.65 mmol)を滴下した。室温で1.5時間攪拌した後、減圧濃縮し、化合物24を得た。精製は行わず次の反応に用いた。
Preparation of 3-trifluoromethyl-3- (ethylenedioxytriisopropylsiloxy-p-phenyl) -3H-diazirine (24 ) A THF solution (15 ml) of compound 5 (300 mg, 1.37 mmol) was cooled to 0 ° C. under an argon atmosphere. Triphenylphosphine (540 mg, 2.06 mmol) was added, and diisopropyl azadicarboxylate (0.41 ml, 2.06 mmol) was added dropwise. The reaction mixture was stirred at 0 ° C. for 10 min, and then compound 13 (333 mg, 1.65 mmol) was added dropwise. After stirring at room temperature for 1.5 hours, the mixture was concentrated under reduced pressure to obtain
3-trifluoromethyl-3-(p-(2'-hydroxyethyl)phenyl)-3H-diazirine (26)の製造
アルゴン雰囲気下、化合物24のTHF(6.9 ml)溶液に、テトラブチルアンモニウムフルオリド(1.08 g, 4.12 mmol)を加えた。反応混合物を室温で3時間攪拌した後、蒸留水を加え、ジエチルエーテルで抽出した。有機層を飽和食塩水で洗浄した後、硫酸マグネシウムで乾燥、減圧濃縮し、化合物26を得た。精製は行わず次の反応に用いた。
Preparation of 3-trifluoromethyl-3- (p- (2'-hydroxyethyl) phenyl) -3H-diazirine (26) Under an argon atmosphere, a solution of
3-trifluoromethyl-3-(p-(2'-(p-toluenesulfonyloxy)ethoxy)phenyl)-3H-diazirine (28)の製造
アルゴン雰囲気下、化合物26のジクロロメタン溶液(0.83 ml)に4-ジメチルアミノピリジン (75 mg, 0.610 mmol)とトリエチルアミン(0.51 ml, 3.68 mmol)を加え、0 ℃に冷却した後、p-トルエンスルホン酸クロリド (345 mg, 2.82 mmol)を加えた。反応混合物を室温で2時間攪拌し、蒸留水を加え、クロロホルムで抽出した。有機層を飽和食塩水で洗浄した後、硫酸マグネシウムで乾燥し、減圧濃縮を行った。残留物をシリカゲルカラムクロマトグラフィー(ヘキサン中9%から25%の酢酸エチル)で精製し化合物28 (0.167 g, 収率69%)を得た。
1H NMR (CDCl3, 400 MHz) d 7.80 (AB-q, J = 8.3 Hz, 1H), 7.31 (AB-q, J = 8.5 Hz, 2H), 7.09 (AB-q, J = 9.0 Hz, 2H), 6.77 (AB-q, J = 8.8 Hz, 2H), 4.36 (t, J = 4.4 Hz, 2H), 4.13 (t, J = 4.4 Hz, 2H), 2.42 (s, 3H), 1.83-1.84 (m, 4H);
13C NMR (CDCl3, 100 MHz) d 158.9, 145.0, 132.8, 129.8, 129.5, 128.4, 128.2, 122.1 (q, J = 274.6 Hz), 116.3, 114.9, 67.8, 65.5, 21.6;
IR (NaCl neat, cm-1) 2959, 2880, 1612, 1520, 1455, 1234, 1192, 1015, 821;
ESI HRMS m/z C19H19F3 N 2O4S [M + Na]+ 403.0602(計算値), found 403.0615(測定値)
3-trifluoromethyl-3- (p- (2 '-(p-toluenesulfonyloxy) ethoxy) phenyl) -3H-diazirine (28) Preparation of 4-dimethylaminopyridine in a dichloromethane solution (0.83 ml) of
1 H NMR (CDCl 3 , 400 MHz) d 7.80 (AB-q, J = 8.3 Hz, 1H), 7.31 (AB-q, J = 8.5 Hz, 2H), 7.09 (AB-q, J = 9.0 Hz, 2H), 6.77 (AB-q, J = 8.8 Hz, 2H), 4.36 (t, J = 4.4 Hz, 2H), 4.13 (t, J = 4.4 Hz, 2H), 2.42 (s, 3H), 1.83- 1.84 (m, 4H);
13 C NMR (CDCl 3 , 100 MHz) d 158.9, 145.0, 132.8, 129.8, 129.5, 128.4, 128.2, 122.1 (q, J = 274.6 Hz), 116.3, 114.9, 67.8, 65.5, 21.6;
IR (NaCl neat, cm -1 ) 2959, 2880, 1612, 1520, 1455, 1234, 1192, 1015, 821;
ESI HRMS m / z C 19 H 19 F 3 N 2 O 4 S [M + Na] + 403.0602 (calculated value), found 403.0615 (measured value)
3-(ethylenedioxy-(p-(p-(2'-(N-(7''-nitrobenzo-2''-oxa-1'',3''-diazol-4''-yl)amino)ethoxy)phenyl)propane-2'-yl)phenyl)-3-trifluoromethyl-3H-diazirine (22)の製造
アルゴン雰囲気下、化合物20(15 mg, 0.035 mmol) のDMF(1 ml)溶液を0 ℃に冷却した後、水素化ナトリウム（油性）(2 mg, 0.053 mmol)を加えた。反応混合物を0 ℃で30分間攪拌した後、化合物28(17 mg, 0.041 mmol)を加え５分間攪拌し、蒸留水を加え、ジエチルエーテルで抽出した。有機層を飽和食塩水で洗浄し、硫酸マグネシウムで乾燥した後、減圧濃縮を行った。残留物を薄層シリカクロマトグラフィー(ヘキサン中50%の酢酸エチル)で精製し、化合物22 (13 mg, 57%)を得た。
1H NMR (CDCl3, 400 MHz) d 8.51 (d, J = 8.8 Hz, 1H), 7.16 (AB-q, J = 8.5 Hz, 2H), 7.15 (AB-q, J = 9.0 Hz, 2H), , 7.13 (AB-q, J = 8.5 Hz, 2H), 6.83 (AB-q, J = 8.8 Hz, 2H), 6.83 (AB-q, J = 8.5 Hz, 2H), 6.82 (AB-q, J = 8.5 Hz, 2H), 6.55 (br-s, 1H), 6.27 (d, J = 8.8 Hz, 1H), 4.28-4.31 (m, 6H), 3.89 (dt, J = 4.9, 5.0 Hz, 2H), 1.63 (s, 6H);
13C NMR (CDCl3, 100 MHz) d 159.7, 156.4, 155.7, 153.0, 144.5, 144.3, 143.4, 136.1, 129.9, 129.3, 128.0, 127.9, 127.8, 122.2 (q, 1JC-F = 274.2 Hz), 116.4, 113.9, 114.9, 66.7, 66.3, 65.2, 43.3, 41.8, 31.0, 29.3(q, 2JC-F = 40.2 Hz);
IR (NaCl neat, cm-1) 2926, 1582, 1510, 1458, 1304, 1182, 1069, 829;
ESI HRMS m/z C33H29F3 N 6O6 [M + Na]+ 685.1998(計算値), 685.1977(測定値)
3- (ethylenedioxy- (p- (p- (2 '-(N- (7``-nitrobenzo-2''-oxa-1'',3''-diazol-4''-yl) amino) ethoxy ) phenyl) propane-2'-yl) phenyl) -3-trifluoromethyl-3H-diazirine (22) Preparation Under argon atmosphere, a solution of compound 20 (15 mg, 0.035 mmol) in DMF (1 ml) was cooled to 0 ° C. After that, sodium hydride (oil) (2 mg, 0.053 mmol) was added. The reaction mixture was stirred at 0 ° C. for 30 min, compound 28 (17 mg, 0.041 mmol) was added and stirred for 5 min, distilled water was added, and the mixture was extracted with diethyl ether. The organic layer was washed with saturated brine, dried over magnesium sulfate, and concentrated under reduced pressure. The residue was purified by thin layer silica chromatography (50% ethyl acetate in hexane) to give compound 22 (13 mg, 57%).
1 H NMR (CDCl 3 , 400 MHz) d 8.51 (d, J = 8.8 Hz, 1H), 7.16 (AB-q, J = 8.5 Hz, 2H), 7.15 (AB-q, J = 9.0 Hz, 2H) ,, 7.13 (AB-q, J = 8.5 Hz, 2H), 6.83 (AB-q, J = 8.8 Hz, 2H), 6.83 (AB-q, J = 8.5 Hz, 2H), 6.82 (AB-q, J = 8.5 Hz, 2H), 6.55 (br-s, 1H), 6.27 (d, J = 8.8 Hz, 1H), 4.28-4.31 (m, 6H), 3.89 (dt, J = 4.9, 5.0 Hz, 2H ), 1.63 (s, 6H);
13 C NMR (CDCl 3 , 100 MHz) d 159.7, 156.4, 155.7, 153.0, 144.5, 144.3, 143.4, 136.1, 129.9, 129.3, 128.0, 127.9, 127.8, 122.2 (q, 1 J CF = 274.2 Hz), 116.4 , 113.9, 114.9, 66.7, 66.3, 65.2, 43.3, 41.8, 31.0, 29.3 (q, 2 J CF = 40.2 Hz);
IR (NaCl neat, cm -1 ) 2926, 1582, 1510, 1458, 1304, 1182, 1069, 829;
ESI HRMS m / z C 33 H 29 F 3 N 6 O 6 [M + Na] + 685.1998 (calculated value), 685.1977 (measured value)
実施例２
本発明の化合物（12）、（19）、（21）、（23）を図１に示す合成スキームにしたがい製造した。
Example 2
Compounds (12), (19), (21) and (23) of the present invention were produced according to the synthetic scheme shown in FIG.
4-(triisopropylsilyloxy)-1-butanol (6)の製造
アルゴン雰囲気下、1, 4−ブタンジオール(4) (2.00 g, 21.8 mmol)のジクロロメタン(4.0 ml)溶液にジイソプロピルエチルアミン(4.0 ml, 23.0 mmol)を滴下した。15分間室温で攪拌したのち、トリイソプロピルシリルクロリド(1.54 ml, 7.18 mmol)を滴下した。反応混合物を2時間室温で攪拌の後、減圧濃縮した。残留物をシリカゲルクロマトグラフィー(ヘキサン中1%から33%の酢酸エチル)により精製し、化合物6 (2.14 g, 収率40%)を得た。
1H NMR (CDCl3, 400 MHz) d 3.75 (t, J = 5.6 Hz, 2H), 3.66 (t, J = 5.6 Hz, 2H), 2.52 (br-s, 1H), 1.62-1.71 (m, 4H), 1.00-1.69 (m, 21H);
13C NMR (CDCl3, 100 MHz) d 63.6, 62.8, 30.3, 30.0, 17.9, 11.9;
IR (NaCl neat, cm-1) 3337, 2946, 2893, 1464, 1385, 789;
ESI HRMS m/z C13H30O2Si [M + Na]+ 269.1913(計算値), found 269.1916(測定値)
Preparation of 4- (triisopropylsilyloxy) -1-butanol (6) Under a argon atmosphere, diisopropylethylamine (4.0 ml, 23.0 mmol) was added to a solution of 1,4-butanediol (4) (2.00 g, 21.8 mmol) in dichloromethane (4.0 ml). ) Was added dropwise. After stirring at room temperature for 15 minutes, triisopropylsilyl chloride (1.54 ml, 7.18 mmol) was added dropwise. The reaction mixture was stirred for 2 hours at room temperature and then concentrated under reduced pressure. The residue was purified by silica gel chromatography (1% to 33% ethyl acetate in hexanes) to give compound 6 (2.14 g, 40% yield).
1 H NMR (CDCl 3 , 400 MHz) d 3.75 (t, J = 5.6 Hz, 2H), 3.66 (t, J = 5.6 Hz, 2H), 2.52 (br-s, 1H), 1.62-1.71 (m, 4H), 1.00-1.69 (m, 21H);
13 C NMR (CDCl 3 , 100 MHz) d 63.6, 62.8, 30.3, 30.0, 17.9, 11.9;
IR (NaCl neat, cm -1 ) 3337, 2946, 2893, 1464, 1385, 789;
ESI HRMS m / z C 13 H 30 O 2 Si [M + Na] + 269.1913 (calculated value), found 269.1916 (measured value)
1-(triisopropylsilyloxy)-4-(p-(2'-(p-(methylenedioxymethyl)phenyl)propane-2'-yl)phenyl)butane (8)の製造
アルゴン雰囲気下、化合物6（100 mg, 0.431 mmol）のTHF(2.0 ml)溶液を0 ℃に冷却した後、トリフェニルホスフィン(170 mg, 0.647 mmol)を加え、ジイソプロピルアザジカルボキシレート(0.12 ml, 0.647 mmol)を滴下した。反応混合物を0 ℃で40分攪拌した後、化合物2を滴下した。室温で3時間攪拌した後、減圧濃縮し化合物8を得た。精製は行わず次の反応に用いた。
1- (triisopropylsilyloxy) -4- (p- (2 '-(p- (methylenedioxymethyl) phenyl) propane-2'-yl) phenyl) butane (8) Preparation of Compound 6 (100 mg, 0.431 mmol ) under argon atmosphere ) In THF (2.0 ml) was cooled to 0 ° C., triphenylphosphine (170 mg, 0.647 mmol) was added, and diisopropyl azadicarboxylate (0.12 ml, 0.647 mmol) was added dropwise. The reaction mixture was stirred at 0 ° C. for 40 minutes, and then compound 2 was added dropwise. The mixture was stirred at room temperature for 3 hours and then concentrated under reduced pressure to obtain
4-(p-(2'-(p-(methylenedioxymethyl)phenyl)propane-2'-yl)phenyl)butanol (10)の製造
アルゴン雰囲気下、化合物8のTHF(4.0 ml)溶液に、テトラブチルアンモニウムフルオリド(338 mg, 1.29 mmol)を加えた。反応混合物を室温で３時間攪拌の後、蒸留水を加え、ジエチルエーテルで抽出した。有機層を飽和食塩水で洗浄後、硫酸マグネシウムで乾燥させ、減圧濃縮した。残留物をシリカゲルカラムクロマトグラフィー(ヘキサン中9%から17%の酢酸エチル)で精製し、化合物10 (88 mg, 収率63%)を得た。
1H NMR (CDCl3, 400 MHz) d 7.14 (AB-q, J = 8.8 Hz, 2H), 7.13 (AB-q, J = 9.0 Hz, 2H), 6.92 (AB-q, J = 9.0 Hz, 2H), 6.85 (AB-q, J = 8.8 Hz, 2H), 5.15 (s, 2H) , 3.98 (t, J = 2.4 Hz, 2H), 3.71 (t, J = 2.4 Hz, 2H), 3.47 (s, 3H), 1.73-1.89 (m, 4H), 1.63 (s, 6H);
13C NMR (CDCl3, 100 MHz) d 155.7, 155.0, 144.4, 143.0, 127.7, 115.6, 113.8, 94.5, 67.7, 62.6, 55.9, 41.7, 31.0, 29.6, 25.9;
IR (NaCl neat, cm-1) 3407, 2963, 2874, 1609, 1510, 1474, 1238, 1182, 1154, 1011, 833;
ESI HRMS m/z C21H28O4 [M + Na]+ 367.1885(計算値), 367.1893(測定値)
4- (p- (2 '-(p- (methylenedioxymethyl) phenyl) propane-2'-yl) phenyl) butanol (10) Preparation of
1 H NMR (CDCl 3 , 400 MHz) d 7.14 (AB-q, J = 8.8 Hz, 2H), 7.13 (AB-q, J = 9.0 Hz, 2H), 6.92 (AB-q, J = 9.0 Hz, 2H), 6.85 (AB-q, J = 8.8 Hz, 2H), 5.15 (s, 2H), 3.98 (t, J = 2.4 Hz, 2H), 3.71 (t, J = 2.4 Hz, 2H), 3.47 ( s, 3H), 1.73-1.89 (m, 4H), 1.63 (s, 6H);
13 C NMR (CDCl 3 , 100 MHz) d 155.7, 155.0, 144.4, 143.0, 127.7, 115.6, 113.8, 94.5, 67.7, 62.6, 55.9, 41.7, 31.0, 29.6, 25.9;
IR (NaCl neat, cm -1 ) 3407, 2963, 2874, 1609, 1510, 1474, 1238, 1182, 1154, 1011, 833;
ESI HRMS m / z C 21 H 28 O 4 [M + Na] + 367.1885 (calculated value), 367.1893 (measured value)
3-trifluoromethyl-3-(1'-(p-(2''-(p-(methylenedioxymethyl)phenyl)propane-2''-yl)phenoxy)butoxy)phenyl-3H-diazirine (12)の製造
アルゴン雰囲気下、化合物10(108 mg, 0.314 mmol) のTHF溶液(1.5 ml)を0 ℃に冷却した後、トリフェニルホスフィン(124 mg, 0.471 mmol)を加え、ジイソプロピルジアザカルボキシレート(0.09 ml, 0.471 mmol)を滴下し、0 ℃で30分間攪拌した後、化合物13(95 mg, 0.471 mmol)を滴下した。反応混合物を室温で12時間攪拌した後、減圧濃縮した。残留物をシリカゲルクロマトグラフィー(ヘキサン中1%から5%の酢酸エチル)で精製し、化合物12(166 mg, 100%)を得た。
1H NMR (CDCl3, 400 MHz) d 7.13 (m, 6H), 6.95 (AB-q, J = 8.8 Hz, 2H), 6.90 (AB-q, J = 9.0 Hz, 2H), 6.85 (AB-q, J = 9.0 Hz, 2H), 5.14 (s, 2H) , 3.98-4.04 (m, 4H), 3.47 (s, 3H), 1.92-1.98 (m, 4H), 1.63 (s, 6H);
13C NMR (CDCl3, 100 MHz) d 160.1, 156.8, 155.1, 144.4, 143.0, 128.1, 127.7, 122.3 (q, 1J C-F = 274.2 Hz), 115.6, 114.9, 113.8, 94.5, 67.7, 67.2, 56.0, 41.7, 31.0, 28.0 (q, 2J C-F = 40.3 Hz), 26.0, 25.9;
IR (NaCl neat, cm-1) 2963, 2081, 1736, 1611, 1582, 1512, 1474, 1236, 1182, 1154, 1011, 829;
ESI HRMS m/z C29H31F3 N 2O4 [M + Na]+ 551.2134(計算値), 551.2149(測定値)
3-trifluoromethyl-3- (1 '-(p- (2''-(p- (methylenedioxymethyl) phenyl) propane-2''-yl) phenoxy) butoxy) phenyl-3H-diazirine (12) Argon atmosphere Then, a THF solution (1.5 ml) of compound 10 (108 mg, 0.314 mmol) was cooled to 0 ° C., triphenylphosphine (124 mg, 0.471 mmol) was added, and diisopropyl diazacarboxylate (0.09 ml, 0.471 mmol). ) Was added dropwise and stirred at 0 ° C. for 30 minutes, and then Compound 13 (95 mg, 0.471 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 12 hours and then concentrated under reduced pressure. The residue was purified by silica gel chromatography (1% to 5% ethyl acetate in hexanes) to give compound 12 (166 mg, 100%).
1 H NMR (CDCl 3 , 400 MHz) d 7.13 (m, 6H), 6.95 (AB-q, J = 8.8 Hz, 2H), 6.90 (AB-q, J = 9.0 Hz, 2H), 6.85 (AB- q, J = 9.0 Hz, 2H), 5.14 (s, 2H), 3.98-4.04 (m, 4H), 3.47 (s, 3H), 1.92-1.98 (m, 4H), 1.63 (s, 6H);
13 C NMR (CDCl 3 , 100 MHz) d 160.1, 156.8, 155.1, 144.4, 143.0, 128.1, 127.7, 122.3 (q, 1 J CF = 274.2 Hz), 115.6, 114.9, 113.8, 94.5, 67.7, 67.2, 56.0 , 41.7, 31.0, 28.0 (q, 2 J CF = 40.3 Hz), 26.0, 25.9;
IR (NaCl neat, cm -1 ) 2963, 2081, 1736, 1611, 1582, 1512, 1474, 1236, 1182, 1154, 1011, 829;
ESI HRMS m / z C 29 H 31 F 3 N 2 O 4 [M + Na] + 551.2134 (calculated value), 551.2149 (measured value)
1-azido-4-(p-(2'-(p-(methylenedioxymethyl)phenyl)propane-2'-yl)phenyl)butane (15)の製造
アルゴン雰囲気下、化合物9(1.55 g, 4.50 mmol) のTHF (20.0 ml) 溶液を0 ℃に冷却した後、トリフェニルホスフィン(2.60 g, 9.90 mmol)を加え、ジイソプロピルアザジカルボキシレート(1.95 ml, 9.90 mmol)を滴下し、0 ℃で5分攪拌した後、ジフェニルホスホリルアジド(2.13 ml, 9.90 mmol)を滴下した。反応混合物を室温で3時間攪拌した後、減圧濃縮し化合物15を得た。精製は行わず次の反応に用いた。
1-azido-4- (p- (2 '-(p- (methylenedioxymethyl) phenyl) propane-2'-yl) phenyl) butane (15) Preparation of Compound 9 (1.55 g, 4.50 mmol) under argon atmosphere After the THF (20.0 ml) solution was cooled to 0 ° C, triphenylphosphine (2.60 g, 9.90 mmol) was added, diisopropyl azadicarboxylate (1.95 ml, 9.90 mmol) was added dropwise, and the mixture was stirred at 0 ° C for 5 minutes. Thereafter, diphenylphosphoryl azide (2.13 ml, 9.90 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 3 hours and then concentrated under reduced pressure to give
1-amino-4-(p-(2'-(p-(methylenedioxymethyl)phenyl)propane-2'-yl)phenyl)butane (17)の製造
アルゴン雰囲気下、化合物15のTHF(40.5 ml)溶液にトリフェニルホスフィン(1.77 g, 6.75 mmol)を加えた。60 ℃に昇温した後、蒸留水(4.5 ml)を加え、60 ℃で3時間攪拌した。反応混合物を室温に冷却した後、蒸留水を加え、酢酸エチルで抽出した。有機層を飽和食塩水で洗浄後、硫酸マグネウシムで乾燥させ、減圧濃縮し化合物17を得た。精製は行わず次の反応に用いた。
Preparation of 1-amino-4- (p- (2 '-(p- (methylenedioxymethyl) phenyl) propane-2'-yl) phenyl) butane (17) In a THF (40.5 ml) solution of
1-(p-(2'-(p-(methylenedioxymethyl)phenyl)propane-2'-yl)phenoxy)-4-(N-(7''-nitrobenzo-2''-oxa-1'',3''-diazol-4''-yl)amino)butane (19)の製造
アルゴン雰囲気下、化合物17のTHF(21.5 ml)溶液に、トリエチルアミン(0.90 ml, 6.75 mmol)を滴下した。0 ℃に冷却した後、7-ニトロ-4-ベンゾフラザニルクロリド(1.03 g, 5.40 mmol)を加えた。反応混合物を室温で30分攪拌した後、飽和塩化アンモニウム水溶液を加え、ジエチルエーテルで抽出した。有機層を飽和食塩水で洗浄し、硫酸マグネシウムで乾燥した後、減圧濃縮した。残留物をシリカゲルクロマトグラフィー(ヘキサン中9%から25%の酢酸エチル)で精製し、化合物19(1.91 g, 収率57%)を得た。
1H NMR (CDCl3, 400 MHz) d 8.48 (d, J = 8.4 Hz, 1H), 7.16 (AB-q, J = 7.3 Hz, 2H), 7.13 (AB-q, J = 7.1 Hz, 2H), 6.93 (AB-q, J = 8.8 Hz, 2H), 6.85 (AB-q, J = 8.8 Hz, 2H), 6.44 (br-s, 1H), 6.19 (d, J = 8.4 Hz, 1H), 5.15 (s, 2H), 4.64 (t, J = 5.6 Hz, 2H), 3.60 (t, J = 6.3 Hz, 2H), 3.47 (s, 3H), 1.95-2.05 (m, 2H), 1.64 (s, 6H);
13C NMR (CDCl3, 100 MHz) d 156.4, 156.1, 144.3, 143.9, 143.8, 143.6, 143.1, 136.4, 127.9, 127.7, 115.9, 113.8, 98.5, 94.5, 67.1, 55.0, 43.7, 41.8, 31.0, 29.7, 26.5, 25.5;
IR (KBr disk, cm-1) 2928, 1626, 1562, 1510, 1439, 1277, 1182, 1152, 1001, 839;
ESI HRMS m/z C27H30 N 4O6 [M − H]- 505.2087(計算値), 505.2092(測定値)
1- (p- (2 '-(p- (methylenedioxymethyl) phenyl) propane-2'-yl) phenoxy) -4- (N- (7''-nitrobenzo-2''-oxa-1'', 3 Preparation of '' -diazol-4 ''-yl) amino) butane (19) Triethylamine (0.90 ml, 6.75 mmol) was added dropwise to a THF (21.5 ml) solution of
1 H NMR (CDCl 3 , 400 MHz) d 8.48 (d, J = 8.4 Hz, 1H), 7.16 (AB-q, J = 7.3 Hz, 2H), 7.13 (AB-q, J = 7.1 Hz, 2H) , 6.93 (AB-q, J = 8.8 Hz, 2H), 6.85 (AB-q, J = 8.8 Hz, 2H), 6.44 (br-s, 1H), 6.19 (d, J = 8.4 Hz, 1H), 5.15 (s, 2H), 4.64 (t, J = 5.6 Hz, 2H), 3.60 (t, J = 6.3 Hz, 2H), 3.47 (s, 3H), 1.95-2.05 (m, 2H), 1.64 (s , 6H);
13 C NMR (CDCl 3 , 100 MHz) d 156.4, 156.1, 144.3, 143.9, 143.8, 143.6, 143.1, 136.4, 127.9, 127.7, 115.9, 113.8, 98.5, 94.5, 67.1, 55.0, 43.7, 41.8, 31.0, 29.7 , 26.5, 25.5;
IR (KBr disk, cm -1 ) 2928, 1626, 1562, 1510, 1439, 1277, 1182, 1152, 1001, 839;
ESI HRMS m / z C 27 H 30 N 4 O 6 [M - H] - 505.2087 ( calcd), 505.2092 (measured value)
1-(p-(2'-(p-hydroxyphenyl)propane-2'-yl)phenoxy)-4-(N-(7''-nitrobenzo-2''-oxa-1'',3''-diazol-4''-yl)amino)butane (21)の製造
化合物19(114 mg, 0.232 mmol) の酢酸 (0.25 ml) 溶液に、2規定塩酸水溶液(0.23 ml, 0.464 mmol)を加えた。反応混合物を室温で3日間攪拌し、蒸留水を加え、ジエチルエーテルで抽出した。有機層を飽和炭酸水素ナトリウム溶液で分液をした後、飽和食塩水で洗浄し、硫酸マグネシウムで乾燥した後、減圧濃縮した。残留物をシリカゲルカラムクロマトグラフィー(ヘキサン中0%から9%の酢酸エチル)で精製し、化合物21(104 mg, 収率77%)を得た。
1H NMR (CDCl3, 400 MHz) d 8.48 (d, J = 8.4 Hz, 1H), 7.15 (AB-q, J = 7.38 Hz, 2H), 7.09 (AB-q, J = 8.8 Hz, 2H), 6.82 (AB-q, J = 8.8 Hz, 2H), 6.73 (AB-q, J = 8.5 Hz, 2H), 6.47 (br-s, 1H), 6.18 (d, J = 8.4 Hz, 1H), 4.04 (t, J = 5.6 Hz, 2H), 3.51-3.60 (m, 2H), 3.47 (s, 3H), 1.94-2.07 (m, 2H), 1.63 (s, 6H);
13C NMR (CDCl3, 100 MHz) d 56.3, 153.3, 144.3, 143.8, 143.7, 143.1, 136.4, 127.9, 127.8, 98.5, 67.0, 43.7, 41.7, 31.0, 29.5, 26.5, 25.5;
IR (KBr disk, cm-1) 3380, 2963, 1620, 1586, 1510, 1445, 1300, 1252, 1184, 1136, 1001, 831;
ESI HRMS m/z C25H26N4O5 [M − H]- 461.1825(計算値), 461.1811(測定値)
1- (p- (2 '-(p-hydroxyphenyl) propane-2'-yl) phenoxy) -4- (N- (7``-nitrobenzo-2''-oxa-1'',3''- To a solution of diazol-4 ″ -yl) amino) butane (21) 19 (114 mg, 0.232 mmol) in acetic acid (0.25 ml), 2N hydrochloric acid aqueous solution (0.23 ml, 0.464 mmol) was added. The reaction mixture was stirred at room temperature for 3 days, distilled water was added, and the mixture was extracted with diethyl ether. The organic layer was separated with a saturated sodium hydrogen carbonate solution, washed with saturated brine, dried over magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0% to 9% ethyl acetate in hexane) to give compound 21 (104 mg, yield 77%).
1 H NMR (CDCl 3 , 400 MHz) d 8.48 (d, J = 8.4 Hz, 1H), 7.15 (AB-q, J = 7.38 Hz, 2H), 7.09 (AB-q, J = 8.8 Hz, 2H) , 6.82 (AB-q, J = 8.8 Hz, 2H), 6.73 (AB-q, J = 8.5 Hz, 2H), 6.47 (br-s, 1H), 6.18 (d, J = 8.4 Hz, 1H), 4.04 (t, J = 5.6 Hz, 2H), 3.51-3.60 (m, 2H), 3.47 (s, 3H), 1.94-2.07 (m, 2H), 1.63 (s, 6H);
13 C NMR (CDCl 3 , 100 MHz) d 56.3, 153.3, 144.3, 143.8, 143.7, 143.1, 136.4, 127.9, 127.8, 98.5, 67.0, 43.7, 41.7, 31.0, 29.5, 26.5, 25.5;
IR (KBr disk, cm -1 ) 3380, 2963, 1620, 1586, 1510, 1445, 1300, 1252, 1184, 1136, 1001, 831;
ESI HRMS m / z C 25 H 26 N 4 O 5 [M - H] - 461.1825 ( calcd), 461.1811 (measured value)
3-trifluoromethyl-3-(p-(4'-(triisopropylsilyloxy)-1'-butoxy)phenyl-3H-diazirine (25)の製造
アルゴン雰囲気下、化合物6(1.00 g, 3.04 mmol) のTHF溶液(15 ml)を0 ℃に冷却した後、トリフェニルホスフィン(1.20 g, 4.56 mmol)を加え、ジイソプロピルアザジカルボキシレート(0.9 ml, 4.56 mmol)を滴下した。0 ℃で10分間攪拌した後、化合物13(0.740 g, 3.65 mmol)を滴下した。反応混合物を室温で1.5時間攪拌した後、減圧濃縮し、化合物25を得た。精製は行わず次の反応に用いた。
Preparation of 3-trifluoromethyl-3- (p- (4 '-(triisopropylsilyloxy) -1'-butoxy) phenyl-3H-diazirine (25) Under argon atmosphere, Compound 6 (1.00 g, 3.04 mmol) in THF (15 ml) was cooled to 0 ° C., triphenylphosphine (1.20 g, 4.56 mmol) was added, and diisopropyl azadicarboxylate (0.9 ml, 4.56 mmol) was added dropwise, and the mixture was stirred at 0 ° C. for 10 min. (0.740 g, 3.65 mmol) was added dropwise, and the reaction mixture was stirred at room temperature for 1.5 hours and then concentrated under reduced pressure to obtain
3-trifluoromethyl-3-(p-(4'-hydroxy-1'-butoxy)phenyl-3H-diazirine (27)の製造
アルゴン雰囲気下、化合物25のTHF(15ml)溶液に、テトラブチルアンモニウムフルオリド(2.40 g, 9.12 mmol)を加えた。反応混合物を室温で3時間攪拌した後、蒸留水を加え、ジエチルエーテルで抽出した。有機層を飽和食塩水で洗浄した後、硫酸マグネシウムで乾燥し減圧濃縮し、化合物27を得た。精製は行わず次の反応に用いた。
Preparation of 3-trifluoromethyl-3- (p- (4'-hydroxy-1'-butoxy) phenyl-3H-diazirine (27) In a THF (15 ml) solution of
3-trifluoromethyl-3-(p-(4'-(p-toluenesulfonyloxy)-1'-butoxy)phenyl-3H-diazirine (29)の製造
アルゴン雰囲気下、化合物27のジクロロメタン溶液(15.2 ml)に、4-ジメチルアミノピリジン(100 mg, 0.808 mmol)とトリエチルアミン(0.68 ml, 4.85 mmol)を加え、0 ℃に冷却した後、p-トルエンスルホニルクロリド(463 mg, 2.42 mmol)を加えた。反応混合物を室温で2時間攪拌し、蒸留水を加え、クロロホルムで抽出した。有機層を飽和食塩水で洗浄した後、硫酸マグネシウムで乾燥し、減圧濃縮を行った。残留物をシリカゲルカラムクロマトグラフィー(9%から25%のヘキサン中、酢酸エチル)で精製し、化合物29 (583 mg, 収率79%)を得た。
1H NMR (CDCl3, 400 MHz) d 7.78 (AB-q, J = 8.3 Hz, 1H), 7.33 (AB-q, J = 8.1 Hz, 2H), 7.12 (AB-q, J = 8.8 Hz, 2H), 6.83 (AB-q, J = 8.8 Hz, 2H), 3.91-4.11 (m, 4H), 2.44 (s, 3H), 1.83-1.84 (m, 4H);
13C NMR (CDCl3, 100 MHz) d 159.8, 144.8, 133.1, 129.8, 129.1, 128.1, 127.9, 122.2 (q, 1JC-F = 274.4 Hz), 116.4, 114.8, 69.9, 67.0, 25.7, 25.2, 21.6;
IR (NaCl neat, cm-1) 2957, 2880, 1613, 1520, 1454, 1236, 1179, 1020, 826;
ESI HRMS m/z C19H19F3 N 2O4S [M + Na]+ 451.0915(計算値), 451.0908(測定値)
Preparation of 3-trifluoromethyl-3- (p- (4 '-(p-toluenesulfonyloxy) -1'-butoxy) phenyl-3H-diazirine (29) Under argon atmosphere, compound 27 in dichloromethane solution (15.2 ml) -Dimethylaminopyridine (100 mg, 0.808 mmol) and triethylamine (0.68 ml, 4.85 mmol) were added, and after cooling to 0 ° C., p-toluenesulfonyl chloride (463 mg, 2.42 mmol) was added. The organic layer was washed with saturated brine, dried over magnesium sulfate, and concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (from 9% to 9%). Purification with 25% ethyl acetate in hexane gave compound 29 (583 mg, 79% yield).
1 H NMR (CDCl 3 , 400 MHz) d 7.78 (AB-q, J = 8.3 Hz, 1H), 7.33 (AB-q, J = 8.1 Hz, 2H), 7.12 (AB-q, J = 8.8 Hz, 2H), 6.83 (AB-q, J = 8.8 Hz, 2H), 3.91-4.11 (m, 4H), 2.44 (s, 3H), 1.83-1.84 (m, 4H);
13 C NMR (CDCl 3 , 100 MHz) d 159.8, 144.8, 133.1, 129.8, 129.1, 128.1, 127.9, 122.2 (q, 1 J CF = 274.4 Hz), 116.4, 114.8, 69.9, 67.0, 25.7, 25.2, 21.6 ;
IR (NaCl neat, cm -1 ) 2957, 2880, 1613, 1520, 1454, 1236, 1179, 1020, 826;
ESI HRMS m / z C 19 H 19 F 3 N 2 O 4 S [M + Na] + 451.0915 (calculated value), 451.0908 (measured value)
3-trifluoromethyl-3-(4'-(p-(2'-(p-(4''-(N-(7'''-nitrobenzo-2'''-oxa-1''',3'''-diazol-4'''-yl)amino)-1'-butoxy)phenyl)propane-2'-yl)phenoxy)butoxy)phenyl-3H-diazirine (23)の製造
アルゴン雰囲気下、化合物21(100 mg, 0.216 mmol) のDMF(3ml)溶液を0 ℃に冷却した後、水素化ナトリウム（油性）(8 mg, 0.324 mmol)を加えた。反応混合物を0 ℃で30分間攪拌し、化合物29(111 mg, 0.259 mmol)を加え５分間攪拌し、蒸留水を加え、ジエチルエーテルで抽出した。飽和食塩水で洗浄し、硫酸マグネシウムで乾燥した後、減圧濃縮を行った。残留物を薄層シリカクロマトグラフィー(ヘキサン中50%の酢酸エチル)で精製し、化合物23(155 mg, 64%)を得た。
1H NMR (CDCl3, 400 MHz) d 8.48 (d, J = 8.5 Hz, 1H), 7.16 (AB-q, J = 9.0 Hz, 2H), 7.12 (AB-q, J = 9.0 Hz, 4H), 6.88 (AB-q, J = 8.5 Hz, 2H), 6.82 (AB-q, J = 8.5 Hz, 2H), 6.78 (AB-q, J = 8.5 Hz, 2H), 6.19 (br-s, 1H), 6.18 (d, J = 8.5 Hz, 1H), 3.98-4.06 (m, 6H), 3.57-3.62 (m, 2H), 1.93-2.04 (m, 8H), 1.64 (s, 6H);
13C NMR (CDCl3, 100 MHz) d 160.3, 156.8, 156.3, 144.3, 143.9, 143.8, 143.7, 143.0, 136.4, 127.8, 127.7, 122.2 (q, 1JC-F = 274.3 Hz), 120.8, 116.4, 114.8, 114.5, 113.8, 98.5, 67.6, 67.3, 67.1, 43.7, 41.7, 31.0, 30.4, 29.7, 29.2 (q, 2JC-F = 43.0 Hz), 26.5, 26.0, 25.9, 25.5;
IR (KBr disk, cm-1)2928, 1613, 1578, 1510, 1470, 1449, 1296, 1182, 1154, 1011, 828;
ESI HRMS m/z C37H37F3 N 6O6 [M + Na]+ 741.2624(計算値), found 741.2641(測定値)
3-trifluoromethyl-3- (4 '-(p- (2'-(p- (4 ''-(N- (7 '''-nitrobenzo-2'''-oxa-1''',3''' -diazol-4 '''-yl) amino) -1'-butoxy) phenyl) propane-2'-yl) phenoxy) butoxy) phenyl-3H-diazirine (23) A solution of 100 mg, 0.216 mmol) in DMF (3 ml) was cooled to 0 ° C., and then sodium hydride (oil) (8 mg, 0.324 mmol) was added. The reaction mixture was stirred at 0 ° C. for 30 min, compound 29 (111 mg, 0.259 mmol) was added and stirred for 5 min, distilled water was added, and the mixture was extracted with diethyl ether. The extract was washed with saturated brine, dried over magnesium sulfate, and concentrated under reduced pressure. The residue was purified by thin layer silica chromatography (50% ethyl acetate in hexane) to give compound 23 (155 mg, 64%).
1 H NMR (CDCl 3 , 400 MHz) d 8.48 (d, J = 8.5 Hz, 1H), 7.16 (AB-q, J = 9.0 Hz, 2H), 7.12 (AB-q, J = 9.0 Hz, 4H) , 6.88 (AB-q, J = 8.5 Hz, 2H), 6.82 (AB-q, J = 8.5 Hz, 2H), 6.78 (AB-q, J = 8.5 Hz, 2H), 6.19 (br-s, 1H ), 6.18 (d, J = 8.5 Hz, 1H), 3.98-4.06 (m, 6H), 3.57-3.62 (m, 2H), 1.93-2.04 (m, 8H), 1.64 (s, 6H);
13 C NMR (CDCl 3 , 100 MHz) d 160.3, 156.8, 156.3, 144.3, 143.9, 143.8, 143.7, 143.0, 136.4, 127.8, 127.7, 122.2 (q, 1 J CF = 274.3 Hz), 120.8, 116.4, 114.8 , 114.5, 113.8, 98.5, 67.6, 67.3, 67.1, 43.7, 41.7, 31.0, 30.4, 29.7, 29.2 (q, 2 J CF = 43.0 Hz), 26.5, 26.0, 25.9, 25.5;
IR (KBr disk, cm -1 ) 2928, 1613, 1578, 1510, 1470, 1449, 1296, 1182, 1154, 1011, 828;
ESI HRMS m / z C 37 H 37 F 3 N 6 O 6 [M + Na] + 741.2624 (calculated value), found 741.2641 (measured value)
実施例３
化合物（11）及び（12）から及び下記スキームにしたがい本発明の化合物（30）及び（31）を製造した。
Example 3
Compounds (30) and (31) of the present invention were produced from compounds (11) and (12) and according to the following scheme.
3-(ethylendioxy-(p-(2'-(p-hydroxyphenyl)propan-2'-yl)phenyl)-3-trifluoromethyl-3H-diazirine (30)の製造
化合物11(80 mg, 0.160 mmol) のアセトニトリル溶液(0.8 ml)に、酢酸 (0.8 ml)、1規定塩酸水溶液(0.80 ml, 0.800 mmol)を加えた。反応混合物を40℃で3時間撹拌し、飽和炭酸水素ナトリウム水溶液を加え、ジエチルエーテルで抽出した。有機層を飽和食塩水で洗浄し、硫酸マグネシウムで乾燥した後、減圧濃縮した。残留物をシリカゲルカラムクロマトグラフィー(ヘキサン中、4%から33%の酢酸エチル)で精製し、化合物30(60 mg, 82%)を得た。
IR (NaCl neat, cm-1) 3405, 1613, 1512, 1458, 1238, 1171, 1071, 829; 1H NMR (CDCl3, 400 MHz) δ 7.14 (m, 4H), 7.07 (AB-q, J = 8.3 Hz, 2H), 6.93 (AB-q, J = 8.6 Hz, 2H), 6.83 (AB-q, J = 8.3 Hz, 2H), 6.71 (AB-q, J = 8.3 Hz, 2H), 5.26 (br-s, 1H), 4.29 (m, 4H), 1.63 (s, 6H); 13C NMR (CDCl3, 100 MHz) δ 159.7, 156.2, 153.4, 143.8, 143.0, 128.1, 127.9, 127.8, 122.2 (q, 1J C-F = 273.8 Hz), 121.3, 115.0, 114.7, 114.0, 66.7, 66.3, 41.7, 31.0, 28.2 (q, 2J C-F = 40.5 Hz); ESI HRMS m/z C25H23F3N2O3 [M − H]- 455.1583(計算値), found 455.1570(測定値)
Preparation of 3- (ethylendioxy- (p- (2 '-(p-hydroxyphenyl) propan-2'-yl) phenyl) -3-trifluoromethyl-3H-diazirine (30) Compound 11 (80 mg, 0.160 Acetic acid (0.8 ml) and 1N aqueous hydrochloric acid (0.80 ml, 0.800 mmol) were added to an acetonitrile solution (0.8 ml) of mmol), and the reaction mixture was stirred at 40 ° C. for 3 hours, and saturated aqueous sodium bicarbonate was added. The organic layer was washed with saturated brine, dried over magnesium sulfate, and concentrated under reduced pressure.The residue was purified by silica gel column chromatography (4% to 33% ethyl acetate in hexane). Compound 30 (60 mg, 82%) was obtained.
IR (NaCl neat, cm -1 ) 3405, 1613, 1512, 1458, 1238, 1171, 1071, 829; 1 H NMR (CDCl 3 , 400 MHz) δ 7.14 (m, 4H), 7.07 (AB-q, J = 8.3 Hz, 2H), 6.93 (AB-q, J = 8.6 Hz, 2H), 6.83 (AB-q, J = 8.3 Hz, 2H), 6.71 (AB-q, J = 8.3 Hz, 2H), 5.26 (br-s, 1H), 4.29 (m, 4H), 1.63 (s, 6H); 13 C NMR (CDCl3, 100 MHz) δ 159.7, 156.2, 153.4, 143.8, 143.0, 128.1, 127.9, 127.8, 122.2 ( q, 1 J CF = 273.8 Hz), 121.3, 115.0, 114.7, 114.0, 66.7, 66.3, 41.7, 31.0, 28.2 (q, 2 J CF = 40.5 Hz); ESI HRMS m / z C 25 H 23 F 3 N 2 O 3 [M - H] - 455.1583 ( calcd), found The 455.1570 (measured value)
3-trifluoromethyl-3-(1'-(p-(2''-(p-hydroxyphenyl)propan-2''-yl)phenoxy)butoxy)phenyl-3H-diazirine (31)の製造
化合物12(53 mg, 0.100 mmol) のアセトニトリル溶液(0.5 ml)に、酢酸 (0.5 ml)、1規定塩酸水溶液(0.50 ml, 0.500 mmol)を加えた。反応混合物を40℃で3時間攪拌し、飽和炭酸水素ナトリウム水溶液を加え、ジエチルエーテルで抽出した。有機層を飽和食塩水で洗浄し、硫酸マグネシウムで乾燥した後、減圧濃縮した。残留物をシリカゲルカラムクロマトグラフィー(ヘキサン中、4%から20%の酢酸エチル)で精製し、化合物31(26 mg, 54%)を得た。
IR (NaCl neat, cm-1) 3364, 1612, 1514, 1474, 1236, 1182, 1053, 829; 1H NMR (CDCl3, 400 MHz) δ 7.13 (m, 4H), 7.09 (AB-q, J = 8.8 Hz, 2H), 6.88 (AB-q, J = 9.0 Hz, 2H), 6.88 (AB-q, J = 8.8 Hz, 2H), 6.72 (AB-q, J = 8.8 Hz, 2H), 4.77 (br-s, 1H), 3.95-4.04 (m, 4H), 1.92-1.98 (m, 4H), 1.62 (s, 6H); 13C NMR (CDCl3, 100 MHz) δ 160.0, 156.8, 153.2, 143.3, 143.2, 128.1, 127.9, 127.7, 122.3 (q, 1J C-F = 269.6 Hz), 120.8, 114.9, 114.7, 113.8, 67.7, 67.3, 41.7, 31.0, 28.1 (q, 2J C-F = 44.7 Hz), 26.0, 25.9;
ESI HRMS m/z C27H27F3N2O3 [M − H]- 483.1896(計算値), found 483.1906(測定値)
3-trifluoromethyl-3- (1 '-(p- (2''-(p-hydroxyphenyl)propan-2''-yl) phenoxy) butoxy) phenyl-3H-diazirine (31) Compound Acetic acid (0.5 ml) and 1N aqueous hydrochloric acid solution (0.50 ml, 0.500 mmol) were added to an acetonitrile solution (0.5 ml) of 12 (53 mg, 0.100 mmol). The reaction mixture was stirred at 40 ° C. for 3 hours, saturated aqueous sodium hydrogen carbonate solution was added, and the mixture was extracted with diethyl ether. The organic layer was washed with saturated brine, dried over magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (4% to 20% ethyl acetate in hexane) to give compound 31 (26 mg, 54%).
IR (NaCl neat, cm -1 ) 3364, 1612, 1514, 1474, 1236, 1182, 1053, 829; 1 H NMR (CDCl 3 , 400 MHz) δ 7.13 (m, 4H), 7.09 (AB-q, J = 8.8 Hz, 2H), 6.88 (AB-q, J = 9.0 Hz, 2H), 6.88 (AB-q, J = 8.8 Hz, 2H), 6.72 (AB-q, J = 8.8 Hz, 2H), 4.77 (br-s, 1H), 3.95-4.04 (m, 4H), 1.92-1.98 (m, 4H), 1.62 (s, 6H); 13 C NMR (CDCl 3 , 100 MHz) δ 160.0, 156.8, 153.2, 143.3, 143.2, 128.1, 127.9, 127.7, 122.3 (q, 1 J CF = 269.6 Hz), 120.8, 114.9, 114.7, 113.8, 67.7, 67.3, 41.7, 31.0, 28.1 (q, 2 J CF = 44.7 Hz), 26.0, 25.9;
ESI HRMS m / z C 27 H 27 F 3 N 2 O 3 [M - H] - 483.1896 ( calcd), found The 483.1906 (measured value)
試験例１
低酸素条件下における赤血球産生調節ホルモン（エリスロポエチン（EPO））の発現に対する、ビスフェノールＡ及び本発明化合物の影響
低酸素感受性因子（HIF-1）には赤血球増殖因子であるエリスロポエチン（EPO）の発現調節因子としての作用が知られている。また、ヒト肝癌細胞（HeP3B）を低酸素状態で培養するとEPOが誘導されてくる現象が知られている。本試験例では、低酸素条件下におけるHeP3B細胞によるEPOの発現に対する、ビスフェノールＡ（BpA）、実施例１で得られた化合物22（BpA-C2；式(4)の化合物）、実施例２で得られた化合物23（BpA-C4；式(5)の化合物）の影響を評価した。
Test example 1
Effect of bisphenol A and the compound of the present invention on the expression of erythropoiesis-regulating hormone (erythropoietin (EPO)) under hypoxic conditions Hypoxia-sensitive factor (HIF-1) regulates the expression of erythropoietin (EPO), a red blood cell growth factor Its action as a factor is known. In addition, it is known that EPO is induced when human hepatoma cells (HeP3B) are cultured under hypoxic conditions. In this test example, bisphenol A (BpA),
HeP3B細胞を10％FCS（仔牛血清）存在下DMEM（Dulbecco's modified Eagle's medium）で培養した。この培養培地に、試験化合物として、200μMのビスフェノールＡ（BpA）、200μMの実施例２で得られた化合物22（BpA-C2）又は200μMの実施例３で得られた化合物23（BpA-C4）を添加した。なお、これらの添加の24時間前にFCS濃度10％の培地をFCS濃度0.1％の培地に変更した。5％の二酸化炭素及び5％の酸素の存在下（低酸素条件下）で6時間培養した。なお、低酸素条件は酸素吸着剤（Anero Pack）を利用して作り出した。また、比較対象として、試験化合物の添加なしでの低酸素条件下での培養、並びに、試験化合物の添加なしで5％の二酸化炭素及び20％の酸素の存在下（通常酸素条件下）での培養も行った。培養後、細胞を回収し、イソゲンを用いてtotal RNAを回収した。このRNAをDNase処理した後、逆転写酵素でcDNAに変換し、下記に示す条件でPCRを行った。
反応混合物：
10 pmolの各プライマー、1.5ユニットのAmpli Taq及び100 ngのcDNA
プライマー：
β−アクチン
5'-CAAGAGATGGCCACGGCTGCT-3'（センス鎖）（配列番号１）
5'-TCCTTCTGCATCCTGTCGGCA-3'（アンチセンス鎖）（配列番号２）
EPO
5'-GCCAGAGGAACTGTCCAGAG-3'（センス鎖）（配列番号３）
5'-TTCTCCAGGTCATCCTGTCC-3'（アンチセンス鎖）（配列番号４）
反応条件：
96℃10分の後、96℃30秒、56℃30秒、72℃1分を１サイクルとし35サイクル
PCR終了後、増幅されたDNAをアガロース電気泳動で分析した。EtBr（エチジウムブロマイド）でDNAを発色させ、その画像をimage analyzer（Atto社 AE6911CX）で取り込み、DNAバンドの濃さを解析ソフトで定量化した。EPOバンドの濃さをβ−アクチンバンドの濃さで割って得られる補正値によって、試験化合物のEPO発現に与える影響を評価した。結果を表１に示す。
HeP3B cells were cultured in DMEM (Dulbecco's modified Eagle's medium) in the presence of 10% FCS (calf serum). In this culture medium, 200 μM of bisphenol A (BpA), 200 μM of compound 22 (BpA-C2) obtained in Example 2 or 200 μM of compound 23 (BpA-C4) obtained in Example 3 were used as test compounds. Was added. In addition, 24 hours before these additions, the medium with an FCS concentration of 10% was changed to a medium with an FCS concentration of 0.1%. The cells were cultured for 6 hours in the presence of 5% carbon dioxide and 5% oxygen (hypoxic conditions). The low oxygen condition was created using an oxygen adsorbent (Anero Pack). In addition, for comparison, the culture was performed under hypoxic conditions without addition of a test compound, and in the presence of 5% carbon dioxide and 20% oxygen without addition of the test compound (normally under oxygen conditions). Culture was also performed. After culturing, the cells were collected, and total RNA was collected using isogen. This RNA was treated with DNase, converted to cDNA with reverse transcriptase, and PCR was performed under the conditions shown below.
Reaction mixture:
10 pmol of each primer, 1.5 units of Ampli Taq and 100 ng cDNA
Primer:
β-actin
5'-CAAGAGATGGCCACGGCTGCT-3 '(sense strand) (SEQ ID NO: 1)
5'-TCCTTCTGCATCCTGTCGGCA-3 '(antisense strand) (SEQ ID NO: 2)
EPO
5'-GCCAGAGGAACTGTCCAGAG-3 '(sense strand) (SEQ ID NO: 3)
5'-TTCTCCAGGTCATCCTGTCC-3 '(antisense strand) (SEQ ID NO: 4)
Reaction conditions:
After 10 minutes at 96 ° C, 35 cycles with 96 ° C for 30 seconds, 56 ° C for 30 seconds and 72 ° C for 1 minute
After completion of PCR, the amplified DNA was analyzed by agarose electrophoresis. DNA was developed with EtBr (ethidium bromide), the image was captured with an image analyzer (Atto AE6911CX), and the intensity of the DNA band was quantified with analysis software. The effect of the test compound on EPO expression was evaluated by a correction value obtained by dividing the density of the EPO band by the density of the β-actin band. The results are shown in Table 1.
BpAの値、BpA-C2の値及びBpA-C4の値は、Hep3Bの低酸素条件の値と比較して有意に減少していた。したがって、BpA-C2、BpA-C4はBpAと同じ生物化学的挙動を示す。特に、BpA-C4はBpAと同程度にEPO発現を抑制していた。 The values of BpA, BpA-C2 and BpA-C4 were significantly reduced compared to the values of Hep3B under hypoxic conditions. Therefore, BpA-C2 and BpA-C4 exhibit the same biochemical behavior as BpA. In particular, BpA-C4 suppressed EPO expression to the same extent as BpA.
配列番号１はβ−アクチンのセンス鎖を示し、配列番号２はβ−アクチンのアンチセンス鎖を示し、配列番号３はエリスロポエチン（EPO）のセンス鎖を示し、配列番号４はEPOのアンチセンス鎖を示す。 SEQ ID NO: 1 shows the sense strand of β-actin, SEQ ID NO: 2 shows the antisense strand of β-actin, SEQ ID NO: 3 shows the sense strand of erythropoietin (EPO), and SEQ ID NO: 4 shows the antisense strand of EPO Indicates.
Claims (5)
又は下記式(3)で表される基
を表す。Ｒ4は−ＯＨ基、上記式(2)で表される基、上記式(3)で表される基又は下記式(4)で表される基
を表す。〕
で表される化合物。 The following formula (1)
Or a group represented by the following formula (3)
Represents. R 4 represents an —OH group, a group represented by the above formula (2), a group represented by the above formula (3), or a group represented by the following formula (4).
Represents. ]
A compound represented by
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