JP2008208039A - Coumarin derivatives and their uses - Google Patents
Coumarin derivatives and their uses Download PDFInfo
- Publication number
- JP2008208039A JP2008208039A JP2007043878A JP2007043878A JP2008208039A JP 2008208039 A JP2008208039 A JP 2008208039A JP 2007043878 A JP2007043878 A JP 2007043878A JP 2007043878 A JP2007043878 A JP 2007043878A JP 2008208039 A JP2008208039 A JP 2008208039A
- Authority
- JP
- Japan
- Prior art keywords
- compound
- solution
- molecular species
- fluorescence
- detection
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 150000001893 coumarin derivatives Chemical class 0.000 title 1
- 150000001875 compounds Chemical class 0.000 claims abstract description 101
- 238000001514 detection method Methods 0.000 claims abstract description 49
- 239000001257 hydrogen Substances 0.000 claims abstract description 20
- 229910052739 hydrogen Inorganic materials 0.000 claims abstract description 20
- 150000002431 hydrogen Chemical class 0.000 claims abstract description 10
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000003795 chemical substances by application Substances 0.000 claims description 22
- 238000000034 method Methods 0.000 claims description 21
- 125000000623 heterocyclic group Chemical group 0.000 claims description 8
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 5
- 239000000126 substance Substances 0.000 abstract description 23
- 150000002632 lipids Chemical class 0.000 abstract description 10
- CFNMUZCFSDMZPQ-GHXNOFRVSA-N 7-[(z)-3-methyl-4-(4-methyl-5-oxo-2h-furan-2-yl)but-2-enoxy]chromen-2-one Chemical compound C=1C=C2C=CC(=O)OC2=CC=1OC/C=C(/C)CC1OC(=O)C(C)=C1 CFNMUZCFSDMZPQ-GHXNOFRVSA-N 0.000 abstract description 8
- 125000000217 alkyl group Chemical group 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 40
- 230000015572 biosynthetic process Effects 0.000 description 30
- 238000003786 synthesis reaction Methods 0.000 description 30
- 241000894007 species Species 0.000 description 29
- 238000006243 chemical reaction Methods 0.000 description 20
- 239000011734 sodium Substances 0.000 description 20
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 15
- 238000002844 melting Methods 0.000 description 15
- 230000008018 melting Effects 0.000 description 15
- FANCTJAFZSYTIS-IQUVVAJASA-N (1r,3s,5z)-5-[(2e)-2-[(1r,3as,7ar)-7a-methyl-1-[(2r)-4-(phenylsulfonimidoyl)butan-2-yl]-2,3,3a,5,6,7-hexahydro-1h-inden-4-ylidene]ethylidene]-4-methylidenecyclohexane-1,3-diol Chemical compound C([C@@H](C)[C@@H]1[C@]2(CCCC(/[C@@H]2CC1)=C\C=C\1C([C@@H](O)C[C@H](O)C/1)=C)C)CS(=N)(=O)C1=CC=CC=C1 FANCTJAFZSYTIS-IQUVVAJASA-N 0.000 description 14
- 229940125904 compound 1 Drugs 0.000 description 14
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 12
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 8
- 239000000523 sample Substances 0.000 description 7
- 125000001424 substituent group Chemical group 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- 239000007864 aqueous solution Substances 0.000 description 5
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N coumarin Chemical compound C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 230000005284 excitation Effects 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- 239000002904 solvent Substances 0.000 description 5
- IZGDXVLRMHXOJV-SFHVURJKSA-N (3s)-4-[2-[2-(4-fluoro-3-methylphenyl)-4-methyl-6-propan-2-ylphenyl]ethyl-hydroxyphosphoryl]-3-hydroxybutanoic acid Chemical compound CC(C)C1=CC(C)=CC(C=2C=C(C)C(F)=CC=2)=C1CCP(O)(=O)C[C@@H](O)CC(O)=O IZGDXVLRMHXOJV-SFHVURJKSA-N 0.000 description 4
- MNIPVWXWSPXERA-IDNZQHFXSA-N (6r,7r)-1-[(4s,5r)-4-acetyloxy-5-methyl-3-methylidene-6-phenylhexyl]-4,7-dihydroxy-6-(11-phenoxyundecanoyloxy)-2,8-dioxabicyclo[3.2.1]octane-3,4,5-tricarboxylic acid Chemical compound C([C@@H](C)[C@H](OC(C)=O)C(=C)CCC12[C@@H]([C@@H](OC(=O)CCCCCCCCCCOC=3C=CC=CC=3)C(O1)(C(O)=O)C(O)(C(O2)C(O)=O)C(O)=O)O)C1=CC=CC=C1 MNIPVWXWSPXERA-IDNZQHFXSA-N 0.000 description 4
- 229940126650 Compound 3f Drugs 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 238000004440 column chromatography Methods 0.000 description 4
- MUTCAPXLKRYEPR-ITWZMISCSA-N methyl (e,3r,5s)-7-[4-bromo-2,3-bis(4-fluorophenyl)-5-propan-2-ylpyrrol-1-yl]-3,5-dihydroxyhept-6-enoate Chemical compound COC(=O)C[C@H](O)C[C@H](O)\C=C\N1C(C(C)C)=C(Br)C(C=2C=CC(F)=CC=2)=C1C1=CC=C(F)C=C1 MUTCAPXLKRYEPR-ITWZMISCSA-N 0.000 description 4
- KDLHZDBZIXYQEI-UHFFFAOYSA-N palladium Substances [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 125000001425 triazolyl group Chemical group 0.000 description 4
- BOOYHBPHFVNWNH-OAHLLOKOSA-N 1-tert-butyl-6-[[(1R)-1-(4-chlorophenyl)ethyl]amino]-5-[(4-fluorophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-one Chemical compound C[C@H](C1=CC=C(C=C1)Cl)NC2=NC3=C(C=NN3C(C)(C)C)C(=O)N2CC4=CC=C(C=C4)F BOOYHBPHFVNWNH-OAHLLOKOSA-N 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- KGPGFQWBCSZGEL-ZDUSSCGKSA-N GSK690693 Chemical compound C=12N(CC)C(C=3C(=NON=3)N)=NC2=C(C#CC(C)(C)O)N=CC=1OC[C@H]1CCCNC1 KGPGFQWBCSZGEL-ZDUSSCGKSA-N 0.000 description 3
- 229960000956 coumarin Drugs 0.000 description 3
- 235000001671 coumarin Nutrition 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- 230000010287 polarization Effects 0.000 description 3
- 150000003384 small molecules Chemical class 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- QMGHHBHPDDAGGO-IIWOMYBWSA-N (2S,4R)-1-[(2S)-2-[[2-[3-[4-[3-[4-[[5-bromo-4-[3-[cyclobutanecarbonyl(methyl)amino]propylamino]pyrimidin-2-yl]amino]phenoxy]propoxy]butoxy]propoxy]acetyl]amino]-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide Chemical compound CN(CCCNC1=NC(NC2=CC=C(OCCCOCCCCOCCCOCC(=O)N[C@H](C(=O)N3C[C@H](O)C[C@H]3C(=O)NCC3=CC=C(C=C3)C3=C(C)N=CS3)C(C)(C)C)C=C2)=NC=C1Br)C(=O)C1CCC1 QMGHHBHPDDAGGO-IIWOMYBWSA-N 0.000 description 2
- WHQUHTXULUACFD-KRWDZBQOSA-N (3s)-4-[[2-(4-fluoro-3-methylphenyl)-4-methyl-6-propan-2-ylphenyl]methoxy-hydroxyphosphoryl]-3-hydroxybutanoic acid Chemical compound CC(C)C1=CC(C)=CC(C=2C=C(C)C(F)=CC=2)=C1COP(O)(=O)C[C@@H](O)CC(O)=O WHQUHTXULUACFD-KRWDZBQOSA-N 0.000 description 2
- PSWDQTMAUUQILQ-UHFFFAOYSA-N 2-[(6-methoxy-4-methylquinazolin-2-yl)amino]-5,6-dimethyl-1h-pyrimidin-4-one Chemical compound N1=C(C)C2=CC(OC)=CC=C2N=C1NC1=NC(=O)C(C)=C(C)N1 PSWDQTMAUUQILQ-UHFFFAOYSA-N 0.000 description 2
- LTYUPYUWXRTNFQ-UHFFFAOYSA-N 5,6-diamino-3',6'-dihydroxyspiro[2-benzofuran-3,9'-xanthene]-1-one Chemical compound C12=CC=C(O)C=C2OC2=CC(O)=CC=C2C11OC(=O)C2=C1C=C(N)C(N)=C2 LTYUPYUWXRTNFQ-UHFFFAOYSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 2
- OZLGRUXZXMRXGP-UHFFFAOYSA-N Fluo-3 Chemical compound CC1=CC=C(N(CC(O)=O)CC(O)=O)C(OCCOC=2C(=CC=C(C=2)C2=C3C=C(Cl)C(=O)C=C3OC3=CC(O)=C(Cl)C=C32)N(CC(O)=O)CC(O)=O)=C1 OZLGRUXZXMRXGP-UHFFFAOYSA-N 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical group C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 2
- 125000003277 amino group Chemical group 0.000 description 2
- 239000012298 atmosphere Substances 0.000 description 2
- OSVHLUXLWQLPIY-KBAYOESNSA-N butyl 2-[(6aR,9R,10aR)-1-hydroxy-9-(hydroxymethyl)-6,6-dimethyl-6a,7,8,9,10,10a-hexahydrobenzo[c]chromen-3-yl]-2-methylpropanoate Chemical compound C(CCC)OC(C(C)(C)C1=CC(=C2[C@H]3[C@H](C(OC2=C1)(C)C)CC[C@H](C3)CO)O)=O OSVHLUXLWQLPIY-KBAYOESNSA-N 0.000 description 2
- XJHCXCQVJFPJIK-UHFFFAOYSA-M caesium fluoride Chemical compound [F-].[Cs+] XJHCXCQVJFPJIK-UHFFFAOYSA-M 0.000 description 2
- 229910001424 calcium ion Inorganic materials 0.000 description 2
- 229940125796 compound 3d Drugs 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000002189 fluorescence spectrum Methods 0.000 description 2
- 125000005843 halogen group Chemical group 0.000 description 2
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 2
- -1 hydrogen ions Chemical class 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 125000002883 imidazolyl group Chemical group 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229910052763 palladium Inorganic materials 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 235000019833 protease Nutrition 0.000 description 2
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 2
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 2
- QKLXBIHSGMPUQS-FGZHOGPDSA-M (3r,5r)-7-[4-(4-fluorophenyl)-2,5-dimethyl-1-phenylpyrrol-3-yl]-3,5-dihydroxyheptanoate Chemical compound CC1=C(CC[C@@H](O)C[C@@H](O)CC([O-])=O)C(C=2C=CC(F)=CC=2)=C(C)N1C1=CC=CC=C1 QKLXBIHSGMPUQS-FGZHOGPDSA-M 0.000 description 1
- VPMIAOSOTOODMY-KJAPKAAFSA-N (4r)-6-[(e)-2-[6-tert-butyl-4-(4-fluorophenyl)-2-propan-2-ylpyridin-3-yl]ethenyl]-4-hydroxyoxan-2-one Chemical compound C([C@H](O)C1)C(=O)OC1/C=C/C=1C(C(C)C)=NC(C(C)(C)C)=CC=1C1=CC=C(F)C=C1 VPMIAOSOTOODMY-KJAPKAAFSA-N 0.000 description 1
- QRDAPCMJAOQZSU-KQQUZDAGSA-N (e)-3-[4-[(e)-3-(3-fluorophenyl)-3-oxoprop-1-enyl]-1-methylpyrrol-2-yl]-n-hydroxyprop-2-enamide Chemical compound C1=C(\C=C\C(=O)NO)N(C)C=C1\C=C\C(=O)C1=CC=CC(F)=C1 QRDAPCMJAOQZSU-KQQUZDAGSA-N 0.000 description 1
- KZPYGQFFRCFCPP-UHFFFAOYSA-N 1,1'-bis(diphenylphosphino)ferrocene Chemical compound [Fe+2].C1=CC=C[C-]1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=C[C-]1P(C=1C=CC=CC=1)C1=CC=CC=C1 KZPYGQFFRCFCPP-UHFFFAOYSA-N 0.000 description 1
- KQZLRWGGWXJPOS-NLFPWZOASA-N 1-[(1R)-1-(2,4-dichlorophenyl)ethyl]-6-[(4S,5R)-4-[(2S)-2-(hydroxymethyl)pyrrolidin-1-yl]-5-methylcyclohexen-1-yl]pyrazolo[3,4-b]pyrazine-3-carbonitrile Chemical compound ClC1=C(C=CC(=C1)Cl)[C@@H](C)N1N=C(C=2C1=NC(=CN=2)C1=CC[C@@H]([C@@H](C1)C)N1[C@@H](CCC1)CO)C#N KQZLRWGGWXJPOS-NLFPWZOASA-N 0.000 description 1
- GOLORTLGFDVFDW-UHFFFAOYSA-N 3-(1h-benzimidazol-2-yl)-7-(diethylamino)chromen-2-one Chemical group C1=CC=C2NC(C3=CC4=CC=C(C=C4OC3=O)N(CC)CC)=NC2=C1 GOLORTLGFDVFDW-UHFFFAOYSA-N 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical group [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical group [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 1
- REDUQXCPUSNJOL-UHFFFAOYSA-N C(C1=CC=CC=C1)NC(CN(C(C1=CC=C(C=C1)C(C)C)=O)CC1=CC=C(C=C1)C(NO)=O)=O Chemical compound C(C1=CC=CC=C1)NC(CN(C(C1=CC=C(C=C1)C(C)C)=O)CC1=CC=C(C=C1)C(NO)=O)=O REDUQXCPUSNJOL-UHFFFAOYSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- CYSWUSAYJNCAKA-FYJFLYSWSA-N ClC1=C(C=CC=2N=C(SC=21)OCC)OC1=CC=C(C=N1)/C=C/[C@H](C)NC(C)=O Chemical compound ClC1=C(C=CC=2N=C(SC=21)OCC)OC1=CC=C(C=N1)/C=C/[C@H](C)NC(C)=O CYSWUSAYJNCAKA-FYJFLYSWSA-N 0.000 description 1
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- 102000005744 Glycoside Hydrolases Human genes 0.000 description 1
- 108010031186 Glycoside Hydrolases Proteins 0.000 description 1
- OPFJDXRVMFKJJO-ZHHKINOHSA-N N-{[3-(2-benzamido-4-methyl-1,3-thiazol-5-yl)-pyrazol-5-yl]carbonyl}-G-dR-G-dD-dD-dD-NH2 Chemical compound S1C(C=2NN=C(C=2)C(=O)NCC(=O)N[C@H](CCCN=C(N)N)C(=O)NCC(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CC(O)=O)C(N)=O)=C(C)N=C1NC(=O)C1=CC=CC=C1 OPFJDXRVMFKJJO-ZHHKINOHSA-N 0.000 description 1
- 101150003085 Pdcl gene Proteins 0.000 description 1
- FZWLAAWBMGSTSO-UHFFFAOYSA-N Thiazole Chemical group C1=CSC=N1 FZWLAAWBMGSTSO-UHFFFAOYSA-N 0.000 description 1
- 206010047139 Vasoconstriction Diseases 0.000 description 1
- YLEIFZAVNWDOBM-ZTNXSLBXSA-N ac1l9hc7 Chemical compound C([C@H]12)C[C@@H](C([C@@H](O)CC3)(C)C)[C@@]43C[C@@]14CC[C@@]1(C)[C@@]2(C)C[C@@H]2O[C@]3(O)[C@H](O)C(C)(C)O[C@@H]3[C@@H](C)[C@H]12 YLEIFZAVNWDOBM-ZTNXSLBXSA-N 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 125000003545 alkoxy group Chemical group 0.000 description 1
- SRVFFFJZQVENJC-IHRRRGAJSA-N aloxistatin Chemical compound CCOC(=O)[C@H]1O[C@@H]1C(=O)N[C@@H](CC(C)C)C(=O)NCCC(C)C SRVFFFJZQVENJC-IHRRRGAJSA-N 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 239000012300 argon atmosphere Substances 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- KSCRVOKQPYZBHZ-IXPOFIJOSA-N benzyl n-[(2s)-1-[[(2s)-1-[[(2s)-1-(1,3-benzothiazol-2-yl)-1-oxo-3-[(3s)-2-oxopyrrolidin-3-yl]propan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]carbamate Chemical compound N([C@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C[C@H]1C(NCC1)=O)C(=O)C=1SC2=CC=CC=C2N=1)C(C)C)C(=O)OCC1=CC=CC=C1 KSCRVOKQPYZBHZ-IXPOFIJOSA-N 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 125000001309 chloro group Chemical group Cl* 0.000 description 1
- 229940126086 compound 21 Drugs 0.000 description 1
- 229940126214 compound 3 Drugs 0.000 description 1
- 229940125877 compound 31 Drugs 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 125000000332 coumarinyl group Chemical group O1C(=O)C(=CC2=CC=CC=C12)* 0.000 description 1
- KFGVRWGDTLZAAO-UHFFFAOYSA-N cyclopenta-1,3-diene dicyclohexyl(cyclopenta-1,3-dien-1-yl)phosphane iron(2+) Chemical compound [Fe++].c1cc[cH-]c1.C1CCC(CC1)P(C1CCCCC1)c1ccc[cH-]1 KFGVRWGDTLZAAO-UHFFFAOYSA-N 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 238000001506 fluorescence spectroscopy Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 229910052731 fluorine Inorganic materials 0.000 description 1
- 125000001153 fluoro group Chemical group F* 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- UKVIEHSSVKSQBA-UHFFFAOYSA-N methane;palladium Chemical compound C.[Pd] UKVIEHSSVKSQBA-UHFFFAOYSA-N 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- QAPTWHXHEYAIKG-RCOXNQKVSA-N n-[(1r,2s,5r)-5-(tert-butylamino)-2-[(3s)-2-oxo-3-[[6-(trifluoromethyl)quinazolin-4-yl]amino]pyrrolidin-1-yl]cyclohexyl]acetamide Chemical compound CC(=O)N[C@@H]1C[C@H](NC(C)(C)C)CC[C@@H]1N1C(=O)[C@@H](NC=2C3=CC(=CC=C3N=CN=2)C(F)(F)F)CC1 QAPTWHXHEYAIKG-RCOXNQKVSA-N 0.000 description 1
- 239000012044 organic layer Substances 0.000 description 1
- 125000002971 oxazolyl group Chemical group 0.000 description 1
- PIBWKRNGBLPSSY-UHFFFAOYSA-L palladium(II) chloride Chemical compound Cl[Pd]Cl PIBWKRNGBLPSSY-UHFFFAOYSA-L 0.000 description 1
- 125000003373 pyrazinyl group Chemical group 0.000 description 1
- 125000003226 pyrazolyl group Chemical group 0.000 description 1
- PBMFSQRYOILNGV-UHFFFAOYSA-N pyridazine Chemical group C1=CC=NN=C1 PBMFSQRYOILNGV-UHFFFAOYSA-N 0.000 description 1
- 125000000714 pyrimidinyl group Chemical group 0.000 description 1
- 125000000168 pyrrolyl group Chemical group 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 235000010288 sodium nitrite Nutrition 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 1
- 230000025033 vasoconstriction Effects 0.000 description 1
Landscapes
- Pyrane Compounds (AREA)
- Plural Heterocyclic Compounds (AREA)
Abstract
【課題】機能性蛍光物質として、特定の分子種の検出や溶液の脂溶性の高さの検出等に好適に利用可能な、新規なクマリン誘導体を提供する。
【解決手段】下記一般式（１）で表される化合物。
〔式中、Ｒ１、Ｒ２、Ｒ３、Ｒ４、Ｒ５は水素又はＣＯＯＲ６等、ＸはＮＲ６Ｒ７を示し、Ｒ６、Ｒ７は水素、アルキル基等を示す。〕
【選択図】なしThe present invention provides a novel coumarin derivative that can be suitably used as a functional fluorescent substance for detection of a specific molecular species, detection of lipid solubility of a solution, and the like.
A compound represented by the following general formula (1):
[Wherein, R 1 , R 2 , R 3 , R 4 , R 5 represent hydrogen or COOR 6 or the like, X represents NR 6 R 7 , R 6 or R 7 represents hydrogen, an alkyl group or the like. ]
[Selection figure] None
Description
本発明は、新規なクマリン誘導体及びその蛍光物質としての用途に関する。 The present invention relates to a novel coumarin derivative and its use as a fluorescent substance.
特定の機能を持った蛍光物質は、多くの分野の科学研究において非常に有用である（非特許文献１〜２）。例えば、特定の分子種（イオン、小分子、酵素等）と結合又は反応することによってその蛍光特性（励起波長、蛍光波長、蛍光強度等）が変化する機能を持った蛍光分子は、その蛍光特性の変化から、分子種の濃度や活性を見積もることができる。
例えば、従来の蛍光物質として、カルシウムイオンと結合することによって蛍光強度が大きく変化するＦｌｕｏ−３（非特許文献３）や、一酸化窒素と反応することによって蛍光強度が増大するＤＡＦ−２（非特許文献４）が知られており（下記反応式参照）、このようなＦｌｕｏ−３やＤＡＦ−２は、それぞれカルシウム蛍光センサーや一酸化窒素蛍光センサーとして有用である。
A fluorescent substance having a specific function is very useful in scientific research in many fields (Non-Patent Documents 1 and 2). For example, a fluorescent molecule having the function of changing its fluorescence characteristics (excitation wavelength, fluorescence wavelength, fluorescence intensity, etc.) by binding or reacting with a specific molecular species (ion, small molecule, enzyme, etc.) From these changes, the concentration and activity of molecular species can be estimated.
For example, as a conventional fluorescent substance, Fluo-3 (Non-patent Document 3) whose fluorescence intensity changes greatly by binding to calcium ions, or DAF-2 (Non-Patent Document 3) whose fluorescence intensity increases by reacting with nitric oxide. Patent Document 4) is known (see the following reaction formula), and such Fluo-3 and DAF-2 are useful as a calcium fluorescence sensor and a nitric oxide fluorescence sensor, respectively.
また、このような蛍光物質は、例えば生命科学の研究では、測定対象の分子種の濃度や活性の時空間的な変化を、生きた生体、組織、細胞を用いて解析することができるため、生体内における分子種の機能を解析する為にも有用である。 In addition, for example, in life science research, such fluorescent substances can be analyzed using living organisms, tissues, and cells for spatiotemporal changes in the concentration and activity of the molecular species to be measured. It is also useful for analyzing the function of molecular species in vivo.
このような有用性の高さから、現在も更に新たな機能性蛍光物質の開発が求められている。 Due to such high utility, development of new functional fluorescent materials is still required.
本発明は、前記諸問題を解決し、以下の目的を達成することを課題とする。即ち、本発明は、機能性蛍光物質として、特定の分子種の検出や溶液の脂溶性の高さの検出等に好適に利用可能な、新規なクマリン誘導体を提供することを目的とする。 An object of the present invention is to solve the above problems and achieve the following object. That is, an object of the present invention is to provide a novel coumarin derivative that can be suitably used as a functional fluorescent substance for detection of a specific molecular species, detection of lipid solubility in a solution, and the like.
本発明者らは、機能性蛍光物質を開発するための方法論として、多種類の候補蛍光物質からなるライブラリー構築が有用であると考えた。そこで本発明者らは、図１に示すように、単一の原料（蛍光団）から同様の反応条件にて多種類の化合物を効率よく得るというコンビナトリアル合成法を蛍光物質に適用し、クマリンを基本骨格とした蛍光物質ライブラリーの構築を行っている。 The present inventors considered that construction of a library composed of many kinds of candidate fluorescent substances is useful as a methodology for developing functional fluorescent substances. Therefore, as shown in FIG. 1, the present inventors applied a combinatorial synthesis method for efficiently obtaining many kinds of compounds under the same reaction conditions from a single raw material (fluorophore) to a fluorescent substance, We are constructing a fluorescent substance library with a basic skeleton.
クマリンの構造、並びに置換位置は以下の通りである。クマリンに蛍光性を持たせるために、通常、７位には水酸基、アミノ基、アルコキシル基等の電子供与性基が導入される。 The structure of coumarin and the substitution position are as follows. In order to give coumarin fluorescence, an electron donating group such as a hydroxyl group, an amino group or an alkoxyl group is usually introduced at the 7-position.
これまでに、クマリンの３位に様々な置換基を導入した蛍光物質ライブラリーが報告されていたが（Ｓｃｈｉｅｄｅｌ，Ｍ．−Ｓ．；Ｂｒｉｅｈｅｎ，Ｃ．Ａ．；Ｂａｕｅｒｌｅ，Ｐ．Ａｎｇｅｗ．Ｃｈｅｍ．Ｉｎｔ．Ｅｄ．２００１，４０，４６７７−４６８０．又はＳｉｌｖａｋｕｍａｒ，Ｋ．；Ｘｉｅ，Ｆ．；Ｃａｓｈ，Ｂ．Ｍ．；Ｌｏｎｇ，Ｓ．；Ｂａｒｎｈｉｌｌ，Ｈ．Ｎ．；Ｗａｎｇ，Ｑ．Ｏｒｇ．Ｌｅｔｔ．２００４，６，４６０３−４６０６．参照）、これに対し、本発明者らは今回、クマリンの５，６，８位に着目し、芳香環、トリアゾール環を含む置換基を効率的に導入する合成法の開発を行った。
その結果、本発明者らは、特定の分子種を検出し得るプローブ（蛍光センサー分子）や、環境応答型蛍光分子等として好適に利用可能な新規クマリン誘導体を見出し、本発明の完成に至った。
So far, a fluorescent substance library in which various substituents are introduced at the 3-position of coumarin has been reported (Schiedel, MS; Briehen, CA; Bauerle, P. Angew. Chem.). Int.Ed. 2001, 40, 4679-4680. Or Silvakumar, K.; Xie, F.; Cash, B.M.; Long, S.; Barnhill, H.N .; Wang, Q. Org.Lett. 2004, 6, 4603-4606.) On the other hand, the present inventors focused on the 5, 6 and 8 positions of coumarin, and this time, the synthesis which introduces the substituent which includes the aromatic ring and the triazole ring efficiently The law was developed.
As a result, the present inventors have found a novel coumarin derivative that can be suitably used as a probe (fluorescent sensor molecule) capable of detecting a specific molecular species, an environmentally responsive fluorescent molecule, and the like, and has completed the present invention. .
本発明は、本発明者らの前記知見に基づくものであり、前記課題を解決するための手段としては、以下の通りである。即ち、
＜１＞ 下記一般式（１）で表されることを特徴とする化合物である。
＜２＞ 下記構造式（３ａ）〜（３ｏ）の少なくともいずれかで表される前記＜１＞に記載の化合物である。
＜３＞ 前記＜１＞から＜２＞のいずれかに記載の化合物を１種以上含むことを特徴とする特定の分子種の検出剤である。
＜４＞ 前記＜１＞から＜２＞のいずれかに記載の化合物を１種以上用いることを特徴とする特定の分子種の検出方法である。
＜５＞ 前記＜１＞から＜２＞のいずれかに記載の化合物を１種以上含むことを特徴とする溶液の脂溶性の高さの検出剤である。
＜６＞ 前記＜１＞から＜２＞のいずれかに記載の化合物を１種以上用いることを特徴とする溶液の脂溶性の高さの検出方法である。
The present invention is based on the above findings of the present inventors, and means for solving the above problems are as follows. That is,
<1> A compound represented by the following general formula (1).
<2> The compound according to <1>, which is represented by at least one of the following structural formulas (3a) to (3o).
<3> A detection agent for a specific molecular species comprising one or more compounds according to any one of <1> to <2>.
<4> A method for detecting a specific molecular species, wherein one or more compounds according to any one of <1> to <2> are used.
<5> A lipid-soluble detection agent for a solution, comprising one or more compounds according to any one of <1> to <2>.
<6> A method for detecting the high fat solubility of a solution, comprising using one or more compounds according to any one of <1> to <2>.
本発明によれば、前記従来における諸問題を解決し、前記目的を達成することができ、機能性蛍光物質として、特定の分子種の検出や溶液の脂溶性の高さの検出等に好適に利用可能な、新規なクマリン誘導体を提供することができる。 According to the present invention, the conventional problems can be solved and the object can be achieved, and as a functional fluorescent substance, it is suitable for detection of specific molecular species, detection of high lipid solubility of a solution, and the like. A novel coumarin derivative that can be used can be provided.
（化合物）
本発明の化合物は、下記一般式（１）で表されることを特徴とする新規クマリン誘導体である。
(Compound)
The compound of the present invention is a novel coumarin derivative represented by the following general formula (1).
＜一般式（１）＞
前記一般式（１）中、Ｒ１は水素又はＣＯＯＲ６を示し；Ｒ２は水素、ＣＨ３、ＣＦ３又はＣＨ２ＣＯＯＲ６を示し；ＸはＯＲ６又はＮＲ６Ｒ７を示し；Ｒ３、Ｒ４及びＲ５はそれぞれ独立に水素、又は下記一般式（２）：
<General formula (1)>
In the general formula (1), R 1 represents hydrogen or COOR 6 ; R 2 represents hydrogen, CH 3 , CF 3 or CH 2 COOR 6 ; X represents OR 6 or NR 6 R 7 ; R 3 , R 4 and R 5 are each independently hydrogen or the following general formula (2):
前記Ｙが、それ（Ｙ）が結合するベンゼン環の隣接する２つの炭素と共に複素環を形成する場合の前記複素環としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、ピロール環、イミダゾール環、ピラゾール環、トリアゾール環、チアゾール環、オキサゾール環、ピリジン環、ピリダジン環、ピリミジン環、ピラジン環などが挙げられる。
これらの中でも、前記複素環としてはトリアゾール環が好ましい。
The heterocyclic ring in the case where Y forms a heterocyclic ring with two adjacent carbons of the benzene ring to which it is bonded is not particularly limited and can be appropriately selected according to the purpose. Pyrrole ring, imidazole ring, pyrazole ring, triazole ring, thiazole ring, oxazole ring, pyridine ring, pyridazine ring, pyrimidine ring, pyrazine ring and the like.
Among these, the heterocyclic ring is preferably a triazole ring.
前記Ｒ６及び前記Ｒ７において、前記ＣＨ３、Ｃ２Ｈ５又はＣ（＝Ｏ）ＣＨ３が置換基を有する場合の前記置換基としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、ハロゲン原子、フェニル基、イミダゾール環、ピリジン環等の複素環基などが挙げられる。前記置換基は、前記ＣＨ３、Ｃ２Ｈ５又はＣ（＝Ｏ）ＣＨ３の１つ乃至複数の水素と置換される。複数の水素と置換される場合には、同一の置換基で置換してもよく、異なる置換基で置換してもよい。前記ハロゲン原子としては、例えば、フッ素原子、塩素原子、臭素原子、ヨウ素原子などが挙げられる。
なお、前記Ｒ６、前記Ｒ７としては、メチル（ＣＨ３）基、エチル（Ｃ２Ｈ５）基が好ましい。
In R 6 and R 7 , the substituent when CH 3 , C 2 H 5 or C (═O) CH 3 has a substituent is not particularly limited and is appropriately selected depending on the purpose. Examples thereof include a heterocyclic group such as a halogen atom, a phenyl group, an imidazole ring, and a pyridine ring. The substituent is substituted with one or more hydrogens of the CH 3 , C 2 H 5 or C (═O) CH 3 . When a plurality of hydrogen atoms are substituted, they may be substituted with the same substituent or may be substituted with different substituents. Examples of the halogen atom include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.
Incidentally, the R 6, as the R 7 is methyl (CH 3) group, ethyl (C 2 H 5) group.
＜具体例＞
前記一般式（１）で表される化合物の具体例としては、例えば下記構造式（３ａ）〜（３ｏ）の少なくともいずれかで表される化合物などが好適に挙げられる。
<Specific example>
Specific examples of the compound represented by the general formula (1) include, for example, compounds represented by at least any one of the following structural formulas (3a) to (3o).
前記構造式（３ａ）〜（３ｏ）中、Ａｒはそれぞれ下記の構造を示す。 In the structural formulas (3a) to (3o), Ar represents the following structure.
＜製造＞
前記化合物（３ａ）〜（３ｏ）の製造方法については、後述する実施例１に詳細かつ具体的に示した。実施例１に記載した方法によれば、一種類の原料化合物（化合物１：実施例１参照）から、同様の反応条件により、多種類の化合物（化合物３ａ〜３ｏ）を効率的に得ることができる。
当業者であれば、実施例１の具体的説明を参照しつつ、原料化合物、反応条件、試薬等を適宜選択し、必要に応じてこれらの方法に適宜の修飾乃至改変を加えることにより、前記一般式（１）に含まれる化合物をいずれも効率的に製造することができる。
<Manufacturing>
About the manufacturing method of the said compounds (3a)-(3o), it showed in detail and concretely in Example 1 mentioned later. According to the method described in Example 1, multiple types of compounds (compounds 3a to 3o) can be efficiently obtained from one kind of raw material compound (compound 1: see Example 1) under the same reaction conditions. it can.
A person skilled in the art can appropriately select raw material compounds, reaction conditions, reagents and the like while referring to the specific description of Example 1, and appropriately modify or modify these methods as necessary. Any compound contained in the general formula (1) can be produced efficiently.
＜用途＞
前記一般式（１）で表される化合物は、蛍光性を有する新規クマリン誘導体であり、その蛍光性を利用して種々の用途に好適に利用可能である。例えば、前記化合物は、後述するような本発明の特定の分子種の検出方法／検出剤や溶液の脂溶性の高さの検出方法／検出剤に好適に利用可能であり、また、これら以外にも様々な用途に好適に利用可能である。
また、例えば前記したような製造方法によれば、一種類の原料化合物から、同様の反応条件により、多種類の一般式（１）で表される化合物を効率的に得ることができる。得られた多種類の化合物は、それぞれが単独で有用であることは勿論であるが、更には得られた多種類の化合物群全体からなる蛍光物質ライブラリーとしても有用である。多種類の化合物を含む蛍光物質ライブラリーを構築しておくことにより、例えば、前記蛍光物質ライブラリーから、所望の機能性を有する蛍光物質（化合物）をより効率的に探索することが可能となる。
<Application>
The compound represented by the general formula (1) is a novel coumarin derivative having fluorescence, and can be suitably used for various applications using the fluorescence. For example, the compound can be suitably used for the detection method / detection agent for the specific molecular species of the present invention and the detection method / detection agent for the lipophilicity of the solution as described later. Can be suitably used for various applications.
For example, according to the production method as described above, various types of compounds represented by the general formula (1) can be efficiently obtained from one kind of raw material compound under the same reaction conditions. Of course, each of the obtained many kinds of compounds is useful alone, but is also useful as a fluorescent substance library composed of the whole obtained many kinds of compounds. By constructing a fluorescent substance library including many kinds of compounds, for example, it becomes possible to more efficiently search for fluorescent substances (compounds) having desired functionality from the fluorescent substance library. .
（特定の分子種の検出方法／特定の分子種の検出剤）
本発明の特定の分子種の検出方法は、前記した本発明の化合物の１種以上を用いることを特徴とする。以下、本発明の特定の分子種の検出方法の説明を通じ、本発明の特定の分子種の検出剤についても説明するものとする。
(Detection method for specific molecular species / detection agent for specific molecular species)
The method for detecting a specific molecular species of the present invention is characterized by using one or more of the aforementioned compounds of the present invention. Hereinafter, the detection agent for the specific molecular species of the present invention will also be described through the description of the method for detecting the specific molecular species of the present invention.
前記したように、本発明の化合物は蛍光性を有する化合物（本明細書中「蛍光物質」と称することがある）であるが、本発明者らは更に、前記化合物が、それらの構造の違いに応じてそれぞれ異なる蛍光特性を有していることを見出した（実施例２（１）参照）。したがって、これらの化合物によれば、前記化合物に結合又は反応することにより前記化合物の構造を変化させる特定の分子種の濃度や活性の強さを、前記化合物の蛍光特性の変化を指標として容易に検出することが可能となる。 As described above, the compound of the present invention is a compound having fluorescence (sometimes referred to as “fluorescent substance” in the present specification). It was found that each had a different fluorescence characteristic (see Example 2 (1)). Therefore, according to these compounds, the concentration and intensity of activity of a specific molecular species that changes the structure of the compound by binding to or reacting with the compound can be easily determined using the change in the fluorescence characteristics of the compound as an index. It becomes possible to detect.
＜特定の分子種＞
前記「特定の分子種」の種類としては、前記化合物に結合又は反応することにより、前記化合物の蛍光特性を変化させる性質を有する分子種であれば、特に制限はなく、目的に応じて適宜選択することができ、例えば、イオン（水素イオン、カルシウムイオンなど）、小分子（一酸化窒素、一重項酸素など）、酵素（ペプチダーゼ、脱アルキル化酵素など）等から適宜選択することができる。
<Specific molecular species>
The type of the “specific molecular species” is not particularly limited as long as it is a molecular species that has the property of changing the fluorescence characteristics of the compound by binding or reacting with the compound, and is appropriately selected according to the purpose. For example, it can be appropriately selected from ions (hydrogen ions, calcium ions, etc.), small molecules (nitrogen monoxide, singlet oxygen, etc.), enzymes (peptidases, dealkylases, etc.) and the like.
＜検出＞
前記特定の分子種の検出方法の実施方法としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、検出対象の試料（例えば、前記特定の分子種を含む試料、前記特定の分子種を含むか否かを評価したい試料等）と、前記化合物とを接触させ、接触前後での前記化合物の蛍光特性の変化を、公知の蛍光測定装置等を用いて測定する方法などが挙げられる。
なお、前記蛍光特性の変化としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、蛍光強度の変化、蛍光波長の変化、励起波長の変化などが挙げられる。
接触前後での前記化合物の蛍光特性の変化の有無や、その変化の程度等から、前記試料中の特定の分子種の濃度や活性の強さ等を、適宜判断することができる（具体例としては、後述する実施例２（１）参照）。
<Detection>
The method for carrying out the method for detecting the specific molecular species is not particularly limited and can be appropriately selected according to the purpose. For example, a sample to be detected (for example, a sample containing the specific molecular species, the specific A sample that is to be evaluated whether or not it contains a molecular species) and the compound, and a method of measuring a change in the fluorescence characteristics of the compound before and after contact using a known fluorescence measuring apparatus or the like. Can be mentioned.
The change in the fluorescence characteristic is not particularly limited and can be appropriately selected according to the purpose. Examples thereof include a change in fluorescence intensity, a change in fluorescence wavelength, and a change in excitation wavelength.
From the presence / absence of the change in the fluorescence characteristics of the compound before and after contact, the degree of the change, etc., the concentration of the specific molecular species in the sample and the strength of activity can be appropriately determined (as a specific example) (See Example 2 (1) described later).
なお、前記検出方法によれば、生きた生体、組織、細胞等を前記試料として用い、これらにおける特定の分子種の濃度や活性の時空間的な変化等を解析することもできる。したがって、前記検出方法は、例えば生命科学の分野において非常に好適である。 According to the detection method, a living organism, tissue, cell, or the like can be used as the sample, and the concentration or activity of a specific molecular species in these can be analyzed in terms of time and space. Therefore, the detection method is very suitable, for example, in the field of life science.
＜検出剤＞
前記特定の分子種の検出方法において、検出対象の試料と接触させる前記化合物としては、前記した本発明の化合物を１種以上含む検出剤（本発明の特定の分子種の検出剤）を使用することが好ましい。前記検出剤に含まれる前記化合物の種類は、検出したい前記分子種の種類に応じて適宜選択することができ、また、前記化合物は１種単独で使用してもよいし、２種以上を併用してもよい。
前記検出剤は、前記化合物そのものであってもよいし、或いは更に適宜その他の成分を含んでいてもよい。前記その他の成分の種類としては、特に制限はなく、目的に応じて適宜選択することができる。また、前記検出剤中の前記化合物と前記その他の成分との含有量比としても、特に制限はなく、目的に応じて適宜選択することができる。
<Detection agent>
In the detection method of the specific molecular species, as the compound to be brought into contact with the sample to be detected, a detection agent (detection agent of the specific molecular species of the present invention) containing one or more of the compounds of the present invention described above is used. It is preferable. The type of the compound contained in the detection agent can be appropriately selected according to the type of the molecular species to be detected, and the compounds may be used alone or in combination of two or more. May be.
The detection agent may be the compound itself or may further contain other components as appropriate. There is no restriction | limiting in particular as a kind of said other component, According to the objective, it can select suitably. Moreover, there is no restriction | limiting in particular as content ratio of the said compound in the said detection agent, and the said other component, According to the objective, it can select suitably.
（溶液の脂溶性の高さの検出方法／溶液の脂溶性の高さの検出剤）
本発明の溶液の脂溶性の高さの検出方法は、前記した本発明の化合物の１種以上を用いることを特徴とする。以下、本発明の溶液の脂溶性の高さの検出方法の説明を通じ、本発明の溶液の脂溶性の高さの検出剤についても説明するものとする。
(Detection method for high fat solubility of solution / detection agent for high fat solubility of solution)
The method for detecting the lipophilicity of the solution of the present invention is characterized by using one or more of the aforementioned compounds of the present invention. Hereinafter, the detection agent for the high fat solubility of the solution of the present invention will also be described through the description of the method for detecting the high fat solubility of the solution of the present invention.
前記したように、本発明の化合物は蛍光性を有する化合物（蛍光物質）であるが、本発明者らは更に、前記化合物が、溶液の脂溶性の高さの違いに応じて蛍光特性が変化する性質を有していることを見出した（実施例２（２）参照）。したがって、これらの化合物によれば、溶液の脂溶性の高さを、前記化合物の蛍光特性の変化を指標として、容易に検出することが可能となる。 As described above, the compound of the present invention is a fluorescent compound (fluorescent substance). However, the present inventors further change the fluorescence characteristics of the compound according to the difference in the lipid solubility of the solution. (See Example 2 (2)). Therefore, according to these compounds, it becomes possible to easily detect the high lipophilicity of the solution using the change in the fluorescence characteristics of the compound as an index.
＜溶液＞
前記「溶液」の種類としては、特に制限はなく、脂溶性の高さを検出したい任意の溶液を、目的に応じて適宜選択することができる。なお、前記「脂溶性」とは、溶媒分子の分極状態を表す指標であり、分極が強い分子は「脂溶性が低い」、分極が弱い分子は「脂溶性が高い」と表される。
<Solution>
There is no restriction | limiting in particular as a kind of said "solution", The arbitrary solutions which want to detect the fat-soluble height can be suitably selected according to the objective. The “fat-soluble” is an index representing the polarization state of solvent molecules. A molecule with strong polarization is expressed as “low in fat solubility” and a molecule with low polarization as “high in fat solubility”.
＜検出＞
前記溶液の脂溶性の高さの検出方法の実施方法としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、検出対象の溶液中での前記化合物の蛍光特性を、公知の蛍光測定装置等を用いて測定し、その蛍光特性を、同様に測定した対照溶液（既に脂溶性の高さがわかっている溶液）中での前記化合物の蛍光特性と比較する方法などが挙げられる。
なお、前記蛍光特性としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、蛍光強度、蛍光波長、励起波長などが挙げられる。
前記検出対象溶液中での蛍光特性と、前記対照溶液中での蛍光特性とを比較することにより、前記検出対象溶液の脂溶性の高さを、前記対照溶液の脂溶性の高さを基準として相対的に判断することができる。
より具体的には、例えば、前記検出対象溶液中での蛍光強度が、前記対照溶液中での蛍光強度よりも高かった場合に、前記検出対象溶液の脂溶性は、前記対照溶液よりも相対的に高いと判断することができ、逆に、前記検出対象溶液中での蛍光強度が、前記対照溶液中での蛍光強度よりも低かった場合に、前記検出対象溶液の脂溶性は、前記対照溶液よりも相対的に低いと判断することができる（実施例２（２）参照）。
<Detection>
The method for detecting the lipid solubility of the solution is not particularly limited and can be appropriately selected depending on the purpose. For example, the fluorescence characteristics of the compound in the solution to be detected are publicly known. And a method of comparing the fluorescence characteristics with the fluorescence characteristics of the compound in a control solution (a solution already known to have high lipid solubility) measured in the same manner.
In addition, there is no restriction | limiting in particular as said fluorescence characteristic, According to the objective, it can select suitably, For example, fluorescence intensity, a fluorescence wavelength, an excitation wavelength, etc. are mentioned.
By comparing the fluorescence characteristics in the detection target solution with the fluorescence characteristics in the control solution, the level of fat solubility of the detection target solution is determined based on the level of fat solubility of the control solution. It can be judged relatively.
More specifically, for example, when the fluorescence intensity in the detection target solution is higher than the fluorescence intensity in the control solution, the fat solubility of the detection target solution is relative to that of the control solution. Conversely, when the fluorescence intensity in the detection target solution is lower than the fluorescence intensity in the control solution, the fat solubility of the detection target solution is the control solution. Therefore, it can be determined that it is relatively lower (see Example 2 (2)).
＜検出剤＞
前記溶液の脂溶性の高さの検出に使用する前記化合物としては、前記した本発明の化合物を１種以上含む検出剤（本発明の溶液の脂溶性の高さの検出剤）を使用することが好ましい。前記検出剤に含まれる前記化合物の種類は、目的に応じて適宜選択することができ、また、前記化合物は１種単独で使用してもよいし、２種以上を併用してもよい。中でも、前記化合物としては、脂溶性の高さの違いによる蛍光強度の違いが大きく、検出がし易い点で、例えば化合物３ｆや化合物３ｈが好ましいと考えられる（実施例２（２）参照）。
前記検出剤は、前記化合物そのものであってもよいし、或いは更に適宜その他の成分を含んでいてもよい。前記その他の成分の種類としては、特に制限はなく、目的に応じて適宜選択することができる。また、前記検出剤中の前記化合物と前記その他の成分との含有量比としても、特に制限はなく、目的に応じて適宜選択することができる。
<Detection agent>
As the compound used for detecting the high fat solubility of the solution, a detection agent containing at least one compound of the present invention described above (detection agent for the high fat solubility of the solution of the present invention) is used. Is preferred. The type of the compound contained in the detection agent can be appropriately selected depending on the purpose, and the compounds may be used alone or in combination of two or more. Among them, as the compound, for example, the compound 3f and the compound 3h are considered to be preferable in that the difference in fluorescence intensity due to the difference in lipid solubility is large and the detection is easy (see Example 2 (2)).
The detection agent may be the compound itself or may further contain other components as appropriate. There is no restriction | limiting in particular as a kind of said other component, According to the objective, it can select suitably. Moreover, there is no restriction | limiting in particular also as content ratio of the said compound in the said detection agent, and the said other component, According to the objective, it can select suitably.
以下に本発明の実施例について説明するが、本発明はこれらの実施例に何ら限定されるものではない。 Examples of the present invention will be described below, but the present invention is not limited to these examples.
（実施例１：化合物３ａ〜３ｏの合成）
実施例１では、下記の合成スキームに従い、本発明の化合物３ａ〜３ｏの合成を行った。
(Example 1: Synthesis of compounds 3a to 3o)
In Example 1, the compounds 3a to 3o of the present invention were synthesized according to the following synthesis scheme.
以下、化合物３ａ〜３ｏの合成手順を詳細に説明する。
なお、上記合成スキームの化合物１は既報の文献（Ｃｏｒｒｉｅ，Ｊ．Ｅ．Ｔ．；Ｍｕｎａｓｉｎｇｈｅ，Ｖ．Ｒ．Ｎ．；Ｒｅｔｔｉｇ，Ｗ．Ｊ．Ｈｅｔｅｒｏｃｙｃｌｉｃ．Ｃｈｅｍ．２０００，３７，１４４７−１４５５．参照）に記された方法により合成した。化合物２ａ〜２ｌは市販品をそのまま用いた。前記市販品はそれぞれ、東京化成工業株式会社、Ａｌｄｒｉｃｈ社、和光純薬工業株式会社のいずれかから適宜入手した。化合物３ｋは化合物３ｍに、化合物３ｌは化合物３ｎに、化合物３ｎは化合物３ｏへと変換し、本発明の化合物１５種類（３ａ〜３ｏ）から成る蛍光物質ライブラリーを構築した。
Hereinafter, the synthesis procedure of compounds 3a to 3o will be described in detail.
In addition, the compound 1 of the said synthetic scheme is the literature (Corrie, JET ;; Munasinghe, VRN; Retig, WJ Heterocyclic. Chem.2000, 37, 1447-1455. ). As compounds 2a to 2l, commercially available products were used as they were. Each of the commercially available products was appropriately obtained from any of Tokyo Chemical Industry Co., Ltd., Aldrich, and Wako Pure Chemical Industries, Ltd. Compound 3k was converted to compound 3m, compound 3l was converted to compound 3n, and compound 3n was converted to compound 3o to construct a fluorescent substance library consisting of 15 compounds (3a to 3o) of the present invention.
＜化合物３ａの合成＞
化合物２ａ（３４．３ｍｇ，０．１６８ｍｍｏｌ）、フッ化セシウム（４３．５ｍｇ，０．２８６ｍｍｏｌ）、１，１’−ビス（ジフェニルホスフィノフェロセン）ジクロロパラジウム（ＩＩ）・塩化メチレン錯体（ＰｄＣｌ２（ｄｐｐｆ）・ＣＨ２Ｃｌ２，９．４ｍｇ，０．０１２ｍｍｏｌ）をＤＭＦ（１ｍｌ）に溶かした溶液に、化合物１（２０．３ｍｇ，０．０５５ｍｍｏｌ）をアルゴン雰囲気下、室温で加え、６０℃に加熱して２時間撹拌した。室温まで冷却した後、反応液を酢酸エチルで希釈し、飽和塩化アンモニウム水溶液、水、飽和食塩水で洗浄した。減圧下で溶媒を留去し、カラムクロマトグラフィーで精製することによって、黄色固体の化合物３ａ（１４．５ｍｇ，７２％）を得た。
融点：１２３−１２４℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・８．４４（ｓ，１Ｈ），７．４０（ｍ，４Ｈ），７．３１（ｍ，１Ｈ），７．２８（ｓ，１Ｈ），６．８５（ｓ，１Ｈ），４．３７（ｑ，Ｊ＝７．１Ｈｚ，２Ｈ），３．０３（ｑ，Ｊ＝７．１Ｈｚ，４Ｈ），１．３７（ｔ，Ｊ＝７．１Ｈｚ，３Ｈ），０．９５（ｔ，Ｊ＝７．０Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１６３．８，１５７．６，１５６．４，１５５．２，１４８．８，１４０．５，１３２．７，１３１．６，１２８．７（２Ｃ），１２８．２（２Ｃ），１２７．３，１１２．８，１１１．１，１０５．７，６１．４，４５．８（２Ｃ），１４．３，１２．１（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ２２Ｈ２３ＮＯ４Ｎａ（Ｍ＋Ｎａ）＋ ３８８．１５２５．Ｆｏｕｎｄ ３８８．１５０２；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ２２Ｈ２３ＮＯ４：Ｃ，７２．３１；Ｈ，６．３４；Ｎ，３．８３．Ｆｏｕｎｄ：Ｃ，７２．３３；Ｈ，６．４５；Ｎ，３．５６．
<Synthesis of Compound 3a>
Compound 2a (34.3 mg, 0.168 mmol), cesium fluoride (43.5 mg, 0.286 mmol), 1,1′-bis (diphenylphosphinoferrocene) dichloropalladium (II) · methylene chloride complex (PdCl 2 ( dppf) · CH 2 Cl 2 , 9.4 mg, 0.012 mmol) in DMF (1 ml) was added Compound 1 (20.3 mg, 0.055 mmol) at room temperature under an argon atmosphere at 60 ° C. Heated and stirred for 2 hours. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate and washed with saturated aqueous ammonium chloride solution, water and saturated brine. The solvent was distilled off under reduced pressure, and the residue was purified by column chromatography to obtain a yellow solid compound 3a (14.5 mg, 72%).
Melting point: 123-124 ° C; 1 H NMR (500 MHz, CDCl 3 ) ... 8.44 (s, 1 H), 7.40 (m, 4 H), 7.31 (m, 1 H), 7.28 ( s, 1H), 6.85 (s, 1H), 4.37 (q, J = 7.1 Hz, 2H), 3.03 (q, J = 7.1 Hz, 4H), 1.37 (t, J = 7.1 Hz, 3H), 0.95 (t, J = 7.0 Hz, 6H); 13 C NMR (125 MHz, CDCl 3 )... 163.8, 157.6, 156.4, 155. 2, 148.8, 140.5, 132.7, 131.6, 128.7 (2C), 128.2 (2C), 127.3, 112.8, 111.1, 105.7, 61. 4,45.8 (2C), 14.3,12.1 (2C ); HRMS (ESI) calcd for C 22 23 NO 4 Na (M + Na ) + 388.1525. Found 388.1502; Anal. Calcd for C 22 H 23 NO 4 : C, 72.31; H, 6.34; N, 3.83. Found: C, 72.33; H, 6.45; N, 3.56.
＜化合物３ｂの合成＞
化合物３ａの合成と同様の方法で、化合物１と化合物２ｂから、収率８５％で黄色粉状物の化合物３ｂを得た。
融点：１７０−１７１℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・８．４２（ｓ，１Ｈ），７．２３（ｓ，１Ｈ），７．２１（ｄ，Ｊ＝８．４Ｈｚ，２Ｈ），６．８０（ｓ，１Ｈ），６．７３（ｄ，Ｊ＝８．３Ｈｚ，２Ｈ），４．３６（ｑ，Ｊ＝７．１Ｈｚ，２Ｈ），３．７６（ｂｒ ｓ，２Ｈ），３．０５（ｑ，Ｊ＝７．１Ｈｚ，４Ｈ），１．３６（ｔ，Ｊ＝７．１Ｈｚ，３Ｈ），０．９５（ｔ，Ｊ＝７．１Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１６３．８，１５７．８，１５６．１，１５５．４，１４８．９，１４５．２，１３２．３，１３１．７，１３０．８，１２９．１（２Ｃ），１１５．５（２Ｃ），１１２．４，１１１．１，１０５．６，６１．４，４５．５（２Ｃ），１４．３，１２．１（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ２２Ｈ２４Ｎ２Ｏ４Ｎａ（Ｍ＋Ｎａ）＋ ４０３．１６３４．Ｆｏｕｎｄ ４０３．１６０８；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ２２Ｈ２４Ｎ２Ｏ４・１／８Ｈ２Ｏ：Ｃ，６９．０５；Ｈ，６．３９；Ｎ，７．３２．Ｆｏｕｎｄ：Ｃ，６９．０５；Ｈ，６．３６；Ｎ，７．３４．
<Synthesis of Compound 3b>
In the same manner as in the synthesis of Compound 3a, Compound 3b as a yellow powder was obtained from Compound 1 and Compound 2b in a yield of 85%.
Melting point: 170-171 ° C .; 1 H NMR (500 MHz, CDCl 3 ) ... 8.42 (s, 1H), 7.23 (s, 1H), 7.21 (d, J = 8.4 Hz, 2H ), 6.80 (s, 1H), 6.73 (d, J = 8.3 Hz, 2H), 4.36 (q, J = 7.1 Hz, 2H), 3.76 (brs, 2H) , 3.05 (q, J = 7.1 Hz, 4H), 1.36 (t, J = 7.1 Hz, 3H), 0.95 (t, J = 7.1 Hz, 6H); 13 C NMR ( 125 MHz, CDCl 3 ) ... 163.8, 157.8, 156.1, 155.4, 148.9, 145.2, 132.3, 131.7, 130.8, 129.1 (2C) 115.5 (2C), 112.4, 111.1, 105.6, 61.4, 45.5 (2C), 14.3, 1 .1 (2C); HRMS (ESI ) calcd for C 22 H 24 N 2 O 4 Na (M + Na) + 403.1634. Found 403.1608; Anal. Calcd for C 22 H 24 N 2 O 4 · 1 / 8H 2 O: C, 69.05; H, 6.39; N, 7.32. Found: C, 69.05; H, 6.36; N, 7.34.
＜化合物３ｃの合成＞
化合物３ａの合成と同様の方法で、化合物１と化合物２ｃから、収率６３％で黄色針状物の化合物３ｃを得た。
融点：１２５−１２６℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・８．４３（ｓ，１Ｈ），７．５７（ｄ，Ｊ＝８．１Ｈｚ，２Ｈ），７．４８（ｓ，１Ｈ），７．３８（ｄ，Ｊ＝８．２Ｈｚ，２Ｈ），７．２５（ｓ，１Ｈ），６．８２（ｓ，１Ｈ），４．３６（ｑ，Ｊ＝７．１Ｈｚ，２Ｈ），３．０３（ｑ，Ｊ＝７．０Ｈｚ，４Ｈ），２．１９（ｓ，３Ｈ），１．３６（ｔ，Ｊ＝７．１Ｈｚ，３Ｈ），０．９５（ｔ，Ｊ＝７．０Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１６８．４，１６３．７，１５７．７，１５６．３，１５５．３，１４８．８，１３７．３，１３６．１，１３２．６，１３１．１，１２８．７（２Ｃ），１２０．０（２Ｃ），１１２．８，１１１．２，１０５．９，６１．５，４５．７（２Ｃ），２４．６，１４．３，１２．１（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ２４Ｈ２６Ｎ２Ｏ５Ｎａ（Ｍ＋Ｎａ）＋ ４４５．１７３９．Ｆｏｕｎｄ ４４５．１７１３；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ２４Ｈ２６Ｎ２Ｏ５：Ｃ，６８．２３；Ｈ，６．２０；Ｎ，６．６３．Ｆｏｕｎｄ：Ｃ，６８．０３；Ｈ，６．２３；Ｎ，６．５７．
<Synthesis of Compound 3c>
In the same manner as the synthesis of compound 3a, yellow needle-like compound 3c was obtained from compound 1 and compound 2c in a yield of 63%.
Melting point: 125-126 ° C .; 1 H NMR (500 MHz, CDCl 3 )... 8.43 (s, 1 H), 7.57 (d, J = 8.1 Hz, 2 H), 7.48 (s, 1 H) ), 7.38 (d, J = 8.2 Hz, 2H), 7.25 (s, 1H), 6.82 (s, 1H), 4.36 (q, J = 7.1 Hz, 2H), 3.03 (q, J = 7.0 Hz, 4H), 2.19 (s, 3H), 1.36 (t, J = 7.1 Hz, 3H), 0.95 (t, J = 7.0 Hz) , 6H); 13 C NMR (125 MHz, CDCl 3 ) 168.4, 163.7, 157.7, 156.3, 155.3, 148.8, 137.3, 136.1, 132. 6, 131.1, 128.7 (2C), 120.0 (2C), 112.8, 111.2, 105.9, 61.5, 4 5.7 (2C), 24.6,14.3,12.1 (2C ); HRMS (ESI) calcd for C 24 H 26 N 2 O 5 Na (M + Na) + 445.1739. Found 445.7133; Anal. Calcd for C 24 H 26 N 2 O 5: C, 68.23; H, 6.20; N, 6.63. Found: C, 68.03; H, 6.23; N, 6.57.
＜化合物３ｄの合成＞
化合物３ａの合成と同様の方法で、化合物１と化合物２ｄから、収率６６％で黄色固体の化合物３ｄを得た。
融点：１６６−１６８℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・８．４２（ｓ，１Ｈ），７．２９（ｄ，Ｊ＝８．７Ｈｚ，２Ｈ），７．２４（ｓ，１Ｈ），６．８０（ｓ，１Ｈ），６．７３（ｄ，Ｊ＝８．４Ｈｚ，２Ｈ），４．３６（ｑ，Ｊ＝７．１Ｈｚ，２Ｈ），３．０６（ｑ，Ｊ＝７．１Ｈｚ，４Ｈ），２．９７（ｓ，６Ｈ），１．３６（ｔ，Ｊ＝７．１Ｈｚ，３Ｈ），０．９６（ｔ，Ｊ＝７．１Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１６３．９，１５７．８，１５５．９，１５５．５，１４９．６，１４８．９，１３２．２，１３１．９，１２８．７（２Ｃ），１２８．２，１１２．５（２Ｃ），１１２．３，１１１．１，１０５．６，６１．３，４５．５（２Ｃ），４０．４（２Ｃ），１４．３，１２．１（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ２４Ｈ２８Ｎ２Ｏ４Ｎａ（Ｍ＋Ｎａ）＋ ４３１．１９４７．Ｆｏｕｎｄ ４３１．１９３０；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ２４Ｈ２８Ｎ２Ｏ４：Ｃ，７０．５７；Ｈ，６．９１；Ｎ，６．８６．Ｆｏｕｎｄ：Ｃ，７０．４１；Ｈ，６．８６；Ｎ，６．８６．
<Synthesis of Compound 3d>
In the same manner as in the synthesis of Compound 3a, Compound 3d as a yellow solid was obtained from Compound 1 and Compound 2d in a yield of 66%.
Melting point: 166-168 ° C .; 1 H NMR (500 MHz, CDCl 3 )... 8.42 (s, 1H), 7.29 (d, J = 8.7 Hz, 2H), 7.24 (s, 1H ), 6.80 (s, 1H), 6.73 (d, J = 8.4 Hz, 2H), 4.36 (q, J = 7.1 Hz, 2H), 3.06 (q, J = 7) .1 Hz, 4H), 2.97 (s, 6H), 1.36 (t, J = 7.1 Hz, 3H), 0.96 (t, J = 7.1 Hz, 6H); 13 C NMR (125 MHz , CDCl 3 ) ... 163.9, 157.8, 155.9, 155.5, 149.6, 148.9, 132.2, 131.9, 128.7 (2C), 128.2 112.5 (2C), 112.3, 111.1, 105.6, 61.3, 45.5 (2C), 40.4 (2C), 14.3,12.1 (2C); HRMS (ESI ) calcd for C 24 H 28 N 2 O 4 Na (M + Na) + 431.1947. Found 431.1930; Anal. Calcd for C 24 H 28 N 2 O 4: C, 70.57; H, 6.91; N, 6.86. Found: C, 70.41; H, 6.86; N, 6.86.
＜化合物３ｅの合成＞
化合物３ａの合成と同様の方法で、化合物１と化合物２ｅから、収率２８％で黄色粉状物の化合物３ｅを得た。
融点：１２３−１２４℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・８．４５（ｓ，１Ｈ），８．２８（ｄ，Ｊ＝８．８Ｈｚ，２Ｈ），７．６４（ｄ，Ｊ＝８．８Ｈｚ，２Ｈ），７．３３（ｓ，１Ｈ），６．９１（ｓ，１Ｈ），４．３８（ｑ，Ｊ＝７．１Ｈｚ，２Ｈ），３．０３（ｑ，Ｊ＝７．１Ｈｚ，４Ｈ），１．３８（ｔ，Ｊ＝７．２Ｈｚ，３Ｈ），０．９８（ｔ，Ｊ＝７．１Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１６３．５，１５７．１，１５６．９，１５４．９，１４８．４，１４７．１，１４７．０，１３２．６，１２９．４，１２９．０（２Ｃ），１２４．２（２Ｃ），１１４．０，１１１．６，１０６．７，６１．７，４６．１（２Ｃ），１４．３，１１．９（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ２２Ｈ２２Ｎ２Ｏ６Ｎａ（Ｍ＋Ｎａ）＋ ４３３．１３７６．Ｆｏｕｎｄ ４３３．１３８１；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ２２Ｈ２２Ｎ２Ｏ６・１／４Ｈ２Ｏ：Ｃ，６３．６８；Ｈ，５．４７；Ｎ，６．７５．Ｆｏｕｎｄ：Ｃ，６３．９０；Ｈ，５．４５；Ｎ，６．４９．
<Synthesis of Compound 3e>
In the same manner as the synthesis of Compound 3a, Compound 3e as a yellow powder was obtained from Compound 1 and Compound 2e in a yield of 28%.
Melting point: 123-124 ° C .; 1 H NMR (500 MHz, CDCl 3 )... 8.45 (s, 1 H), 8.28 (d, J = 8.8 Hz, 2 H), 7.64 (d, J = 8.8 Hz, 2H), 7.33 (s, 1H), 6.91 (s, 1H), 4.38 (q, J = 7.1 Hz, 2H), 3.03 (q, J = 7) .1 Hz, 4H), 1.38 (t, J = 7.2 Hz, 3H), 0.98 (t, J = 7.1 Hz, 6H); 13 C NMR (125 MHz, CDCl 3 ). 5, 157.1, 156.9, 154.9, 148.4, 147.1, 147.0, 132.6, 129.4, 129.0 (2C), 124.2 (2C), 114. 0, 111.6, 106.7, 61.7, 46.1 (2C), 14.3, 11.9 (2C); HRMS (E SI) calcd for C 22 H 22 N 2 O 6 Na (M + Na) + 433.1376. Found 433.1381; Anal. Calcd for C 22 H 22 N 2 O 6 · 1 / 4H 2 O: C, 63.68; H, 5.47; N, 6.75. Found: C, 63.90; H, 5.45; N, 6.49.
＜化合物３ｆの合成＞
化合物３ａの合成と同様の方法で、化合物１と化合物２ｆから、収率７５％で黄色針状物の化合物３ｆを得た。
融点：１１２−１１３℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・８．４４（ｓ，１Ｈ），７．２９（ｄ，Ｊ＝８．６Ｈｚ，２Ｈ），７．２５（ｓ，１Ｈ），６．８８（ｄ，Ｊ＝８．６Ｈｚ，２Ｈ），６．８４（ｓ，１Ｈ），５，１５（ｂｒ ｓ，１Ｈ），４．３７（ｑ，Ｊ＝７．１Ｈｚ，２Ｈ），３．０４（ｑ，Ｊ＝７．１Ｈｚ，４Ｈ），１．３７（ｔ，Ｊ＝７．１Ｈｚ，３Ｈ），０．９６（ｔ，Ｊ＝７．１Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１６３．９，１５８．１，１５６．２，１５５．５，１５５．４，１４９．１，１３２．６，１３２．５，１３１．５，１２９．４（２Ｃ），１１５．８（２Ｃ），１１２．３，１１１．１，１０５．６，６１．５，４５．６（２Ｃ），１４．３，１２．１（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ２２Ｈ２２ＮＯ５（Ｍ−Ｈ）− ３８０．１４９８．Ｆｏｕｎｄ ３８０．１４９１；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ２２Ｈ２３ＮＯ５：Ｃ，６９．２８；Ｈ，６．０８；Ｎ，３．６７．Ｆｏｕｎｄ：Ｃ，６９．０２；Ｈ，６．２０；Ｎ，３．６５．
<Synthesis of Compound 3f>
By a method similar to the synthesis of Compound 3a, Compound 3f as a yellow needle was obtained from Compound 1 and Compound 2f in a yield of 75%.
Melting point: 112-113 ° C .; 1 H NMR (500 MHz, CDCl 3 ) ... 8.44 (s, 1H), 7.29 (d, J = 8.6 Hz, 2H), 7.25 (s, 1H ), 6.88 (d, J = 8.6 Hz, 2H), 6.84 (s, 1H), 5, 15 (brs, 1H), 4.37 (q, J = 7.1 Hz, 2H) , 3.04 (q, J = 7.1 Hz, 4H), 1.37 (t, J = 7.1 Hz, 3H), 0.96 (t, J = 7.1 Hz, 6H); 13 C NMR ( 125 MHz, CDCl 3 ) ... 163.9, 158.1, 156.2, 155.5, 155.4, 149.1, 132.6, 132.5, 131.5, 129.4 (2C) 115.8 (2C), 112.3, 111.1, 105.6, 61.5, 45.6 (2C), 14.3, 1 .1 (2C); HRMS (ESI ) calcd for C 22 H 22 NO 5 (M-H) - 380.1498. Found 380.1491; Anal. Calcd for C 22 H 23 NO 5 : C, 69.28; H, 6.08; N, 3.67. Found: C, 69.02; H, 6.20; N, 3.65.
＜化合物３ｇの合成＞
化合物３ａの合成と同様の方法で、化合物１と化合物２ｇから、収率６３％で黄色針状物の化合物３ｇを得た。
融点：７０−７１℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・８．４３（ｓ，１Ｈ），７．３５（ｄ，Ｊ＝８．８Ｈｚ，２Ｈ），７．２６（ｓ，１Ｈ），６．９３（ｄ，Ｊ＝８．８Ｈｚ，２Ｈ），６．８５（ｓ，１Ｈ），４．３７（ｑ，Ｊ＝７．１Ｈｚ，２Ｈ），３．８４（ｓ，３Ｈ），３．０４（ｑ，Ｊ＝７．１Ｈｚ，４Ｈ），１．３７（ｔ，Ｊ＝７．１Ｈｚ，３Ｈ），０．９６（ｔ，Ｊ＝７．１Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１６３．８，１５９．０，１５７．６，１５６．２，１５５．１，１４８．８，１３２．７，１３２．５，１３１．４，１２９．３（２Ｃ），１１４．２（２Ｃ），１１２．９，１１１．３，１０６．０，６１．５，５５．３，４５．８（２Ｃ），１４．３，１２．１（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ２３Ｈ２５ＮＯ５Ｎａ（Ｍ＋Ｎａ）＋ ４１８．１６３０．Ｆｏｕｎｄ ４１８．１６０８ｒｔ；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ２３Ｈ２５ＮＯ５：Ｃ，６９．８６；Ｈ，６．３７；Ｎ，３．５４．Ｆｏｕｎｄ：Ｃ， ６９．６２；Ｈ，６．４０；Ｎ，３．５４．
<Synthesis of Compound 3g>
By a method similar to the synthesis of Compound 3a, Compound 3g as a yellow needle was obtained from Compound 1 and Compound 2g in a yield of 63%.
Melting point: 70-71 ° C .; 1 H NMR (500 MHz, CDCl 3 )... 8.43 (s, 1H), 7.35 (d, J = 8.8 Hz, 2H), 7.26 (s, 1H) ), 6.93 (d, J = 8.8 Hz, 2H), 6.85 (s, 1H), 4.37 (q, J = 7.1 Hz, 2H), 3.84 (s, 3H), 3.04 (q, J = 7.1 Hz, 4H), 1.37 (t, J = 7.1 Hz, 3H), 0.96 (t, J = 7.1 Hz, 6H); 13 C NMR (125 MHz , CDCl 3 ) ... 163.8, 159.0, 157.6, 156.2, 155.1, 148.8, 132.7, 132.5, 131.4, 129.3 (2C), 114.2 (2C), 112.9, 111.3, 106.0, 61.5, 55.3, 45.8 (2C), 14.3, 1 2.1 (2C); HRMS (ESI ) calcd for C 23 H 25 NO 5 Na (M + Na) + 418.1630. Found 418.1608 rt; Anal. Calcd for C 23 H 25 NO 5 : C, 69.86; H, 6.37; N, 3.54. Found: C, 69.62; H, 6.40; N, 3.54.
＜化合物３ｈの合成＞
化合物３ａの合成と同様の方法で、化合物１と化合物２ｈから、収率５８％で黄色固体の化合物３ｈを得た。
融点：７９−８０℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・８．４４（ｓ，１Ｈ），８．００（ｄ，Ｊ＝８．４Ｈｚ，２Ｈ），７．５５（ｄ，Ｊ＝８．４Ｈｚ，２Ｈ），７．３０（ｓ，１Ｈ），６．８７（ｓ，１Ｈ），４．３７（ｑ，Ｊ＝７．１Ｈｚ，２Ｈ），３．０２（ｑ，Ｊ＝７．１Ｈｚ，４Ｈ），２．６２（ｓ，３Ｈ），１．３７（ｔ，Ｊ＝７．１Ｈｚ，３Ｈ），０．９６（ｔ，Ｊ＝７．１Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１９７．４，１６３．６，１５７．３，１５６．７，１５５．０，１４８．６，１４５．４，１３６．０，１３２．６，１３０．４，１２８．９（２Ｃ），１２８．４（２Ｃ），１１３．４，１１１．３，１０６．２，６１．６，４５．９（２Ｃ），２６．６，１４．３，１２．０（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ２４Ｈ２５ＮＯ５Ｎａ（Ｍ＋Ｎａ）＋ ４３０．１６３０．Ｆｏｕｎｄ ４３０．１６０６；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ２４Ｈ２５ＮＯ５：Ｃ，７０．７４；Ｈ，６．１８；Ｎ，３．４４．Ｆｏｕｎｄ：Ｃ，７０．５５；Ｈ，６．２９；Ｎ，３．４２．
<Synthesis of Compound 3h>
In the same manner as the synthesis of Compound 3a, Compound 3h as a yellow solid was obtained from Compound 1 and Compound 2h in a yield of 58%.
Melting point: 79-80 ° C .; 1 H NMR (500 MHz, CDCl 3 ) ... 8.44 (s, 1H), 8.00 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 8.4 Hz, 2H), 7.30 (s, 1H), 6.87 (s, 1H), 4.37 (q, J = 7.1 Hz, 2H), 3.02 (q, J = 7) .1 Hz, 4H), 2.62 (s, 3H), 1.37 (t, J = 7.1 Hz, 3H), 0.96 (t, J = 7.1 Hz, 6H); 13 C NMR (125 MHz , CDCl 3 ) ... 197.4, 163.6, 157.3, 156.7, 155.0, 148.6, 145.4, 136.0, 132.6, 130.4, 128.9 (2C), 128.4 (2C), 113.4, 111.3, 106.2, 61.6, 45.9 (2C), 26.6 14.3,12.0 (2C); HRMS (ESI ) calcd for C 24 H 25 NO 5 Na (M + Na) + 430.1630. Found 430.1606; Anal. Calcd for C 24 H 25 NO 5 : C, 70.74; H, 6.18; N, 3.44. Found: C, 70.55; H, 6.29; N, 3.42.
＜化合物３ｉの合成＞
化合物３ａの合成と同様の方法で、化合物１と化合物２ｉから、収率４５％で黄色針状物の化合物３ｉを得た。
融点：１７４−１７６℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・８．４４（ｓ，１Ｈ），７．７０（ｄ，Ｊ＝８．３Ｈｚ，２Ｈ），７．５８（ｄ，Ｊ＝８．３Ｈｚ，２Ｈ），７．２９（ｓ，１Ｈ），６．８９（ｓ，１Ｈ），４．３７（ｑ，Ｊ＝７．１Ｈｚ，２Ｈ），３．０１（ｑ，Ｊ＝７．１Ｈｚ，４Ｈ），１．３７（ｔ，Ｊ＝７．１Ｈｚ，３Ｈ），０．９７（ｔ，Ｊ＝７．０Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１６３．５，１５７．２，１５６．８，１５４．９，１４８．５，１４５．２，１３２．６（２Ｃ），１３２．６，１２９．７，１２８．９（２Ｃ），１１８．６，１１３．７，１１１．５，１１１．２，１０６．５，６１．６，４５．９（２Ｃ），１４．３，１１．９（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ２３Ｈ２２Ｎ２Ｏ４Ｎａ（Ｍ＋Ｎａ）＋ ４１３．１４７７．Ｆｏｕｎｄ ４１３．１４６２；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ２３Ｈ２２Ｎ２Ｏ４：Ｃ，７０．７５；Ｈ，５．６８；Ｎ，７．１７．Ｆｏｕｎｄ：Ｃ，７０．６１；Ｈ，５．７５；Ｎ，６．９９．
<Synthesis of Compound 3i>
In the same manner as the synthesis of compound 3a, yellow needle-like compound 3i was obtained from compound 1 and compound 2i in a yield of 45%.
Melting point: 174 to 176 ° C; 1 H NMR (500 MHz, CDCl 3 ) ... 8.44 (s, 1 H), 7.70 (d, J = 8.3 Hz, 2 H), 7.58 (d, J = 8.3 Hz, 2H), 7.29 (s, 1H), 6.89 (s, 1H), 4.37 (q, J = 7.1 Hz, 2H), 3.01 (q, J = 7) .1 Hz, 4H), 1.37 (t, J = 7.1 Hz, 3H), 0.97 (t, J = 7.0 Hz, 6H); 13 C NMR (125 MHz, CDCl 3 ). 5, 157.2, 156.8, 154.9, 148.5, 145.2, 132.6 (2C), 132.6, 129.7, 128.9 (2C), 118.6, 113. 7, 111.5, 111.2, 106.5, 61.6, 45.9 (2C), 14.3, 11.9 (2C); HRMS (ESI) calcd for C 23 H 22 N 2 O 4 Na (M + Na) + 413.1477. Found 413.1462; Anal. Calcd for C 23 H 22 N 2 O 4: C, 70.75; H, 5.68; N, 7.17. Found: C, 70.61; H, 5.75; N, 6.99.
＜化合物３ｊの合成＞
化合物３ａの合成と同様の方法で、化合物１と化合物２ｊから、収率４６％で黄色固体の化合物３ｊを得た。
融点：１１５−１１７℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・８．４４（ｓ，１Ｈ），８．０７（ｄ，Ｊ＝８．２Ｈｚ，２Ｈ），７．５２（ｄ，Ｊ＝８．３Ｈｚ，２Ｈ），７．３１（ｓ，１Ｈ），６．８８（ｓ，１Ｈ），４．３７（ｑ，Ｊ＝７．１Ｈｚ，２Ｈ），３．９３（ｓ，３Ｈ），３．０２（ｑ，Ｊ＝７．１Ｈｚ，４Ｈ），１．３７（ｔ，Ｊ＝７．１Ｈｚ，３Ｈ），０．９６（ｔ，Ｊ＝７．０Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１６６．７，１６３．６，１５７．３，１５６．６，１５５．０，１４８．６，１４５．２，１３２．６，１３０．４，１３０．１（２Ｃ），１２９．１，１２８．２（２Ｃ），１１３．２，１１１．２，１０６．０，６１．５，５２．１，４５．８（２Ｃ），１４．２，１２．０（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ２４Ｈ２５ＮＯ６Ｎａ（Ｍ＋Ｎａ）＋ ４４６．１５８０．Ｆｏｕｎｄ ４４６．１５８３；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ２４Ｈ２５ＮＯ６：Ｃ，６８．０７；Ｈ，５．９５；Ｎ，３．３１．Ｆｏｕｎｄ：Ｃ，６７．７８；Ｈ，５．９４；Ｎ，３．３０．
<Synthesis of Compound 3j>
In the same manner as in the synthesis of Compound 3a, Compound 3j as a yellow solid was obtained from Compound 1 and Compound 2j in a yield of 46%.
Melting point: 115-117 ° C .; 1 H NMR (500 MHz, CDCl 3 ) ... 8.44 (s, 1H), 8.07 (d, J = 8.2 Hz, 2H), 7.52 (d, J = 8.3 Hz, 2H), 7.31 (s, 1H), 6.88 (s, 1H), 4.37 (q, J = 7.1 Hz, 2H), 3.93 (s, 3H), 3.02 (q, J = 7.1 Hz, 4H), 1.37 (t, J = 7.1 Hz, 3H), 0.96 (t, J = 7.0 Hz, 6H); 13 C NMR (125 MHz , CDCl 3 ) ... 166.7, 163.6, 157.3, 156.6, 155.0, 148.6, 145.2, 132.6, 130.4, 130.1 (2C), 129.1, 128.2 (2C), 113.2, 111.2, 106.0, 61.5, 52.1, 45.8 (2C ), 14.2,12.0 (2C); HRMS (ESI) calcd for C 24 H 25 NO 6 Na (M + Na) + 446.1580. Found 446.1583; Anal. Calcd for C 24 H 25 NO 6 : C, 68.07; H, 5.95; N, 3.31. Found: C, 67.78; H, 5.94; N, 3.30.
＜化合物３ｋの合成＞
化合物３ａの合成と同様の方法で、化合物１と化合物２ｋから、収率６３％で黄色固体の化合物３ｋを得た。
融点：１２１−１２３℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・８．４４（ｓ，１Ｈ），８．１１（ｄ，Ｊ＝８．４Ｈｚ，２Ｈ），７．５２（ｄ，Ｊ＝８．５Ｈｚ，２Ｈ），７．４６（ｄ，Ｊ＝７．０Ｈｚ，２Ｈ），７．３９（ｍ，２Ｈ），７．３４（ｍ，１Ｈ），７．３０（ｓ，１Ｈ），６．８６（ｓ，１Ｈ），５．３７（ｓ，２Ｈ），４．３７（ｑ，Ｊ＝７．１Ｈｚ，２Ｈ），３．０１（ｑ，Ｊ＝７．１Ｈｚ，４Ｈ），１．３７（ｔ，Ｊ＝７．１Ｈｚ，３Ｈ），０．９５（ｔ，Ｊ＝７．１Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１６６．１，１６３．６，１５７．４，１５６．７，１５５．０，１４８．６，１４５．３，１３６．０，１３２．６，１３０．４，１３０．２（２Ｃ），１２９．１，１２８．６（２Ｃ），１２８．３３，１２８．２７（２Ｃ），１２８．２（２Ｃ），１１３．３，１１１．３，１０６．１，６６．８，６１．５，４５．９（２Ｃ），１４．３，１２．０（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ３０Ｈ２９ＮＯ６Ｎａ（Ｍ＋Ｎａ）＋ ５２２．１８９３．Ｆｏｕｎｄ ５２２．１８８８；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ３０Ｈ２９ＮＯ６・１／８Ｈ２Ｏ：Ｃ，７１．８０；Ｈ，５．８８；Ｎ，２．７９．Ｆｏｕｎｄ：Ｃ，７１．７０；Ｈ，６．０１；Ｎ，２．８１．
<Synthesis of Compound 3k>
In the same manner as the synthesis of Compound 3a, Compound 3k as a yellow solid was obtained from Compound 1 and Compound 2k in a yield of 63%.
Melting point: 121-123 ° C .; 1 H NMR (500 MHz, CDCl 3 ) ... 8.44 (s, 1H), 8.11 (d, J = 8.4 Hz, 2H), 7.52 (d, J = 8.5 Hz, 2H), 7.46 (d, J = 7.0 Hz, 2H), 7.39 (m, 2H), 7.34 (m, 1H), 7.30 (s, 1H), 6.86 (s, 1H), 5.37 (s, 2H), 4.37 (q, J = 7.1 Hz, 2H), 3.01 (q, J = 7.1 Hz, 4H), 1. 37 (t, J = 7.1 Hz, 3H), 0.95 (t, J = 7.1 Hz, 6H); 13 C NMR (125 MHz, CDCl 3 ) 166.1, 163.6, 157. 4, 156.7, 155.0, 148.6, 145.3, 136.0, 132.6, 130.4, 130.2 (2C), 129. 1, 128.6 (2C), 128.33, 128.27 (2C), 128.2 (2C), 113.3, 111.3, 106.1, 66.8, 61.5, 45.9 (2C), 14.3,12.0 (2C) ; HRMS (ESI) calcd for C 30 H 29 NO 6 Na (M + Na) + 522.1893. Found 522.1888; Anal. Calcd for C 30 H 29 NO 6 · 1 / 8H 2 O: C, 71.80; H, 5.88; N, 2.79. Found: C, 71.70; H, 6.01; N, 2.81.
＜化合物３ｌの合成＞
化合物３ａの合成と同様の方法で、化合物１と化合物２ｌから、収率６２％で黄色粉状物の化合物３ｌを得た。
融点：１８７−１８８℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・８．４４（ｓ，１Ｈ），８．２３（ｄ，Ｊ＝２．１Ｈｚ，１Ｈ），７．５２（ｄｄ，Ｊ＝８．６，２．１Ｈｚ，１Ｈ），７．３１（ｓ，１Ｈ），６．８９（ｓ，１Ｈ），６．８６（ｄ，Ｊ＝８．６Ｈｚ，１Ｈ），６．２１（ｂｒ ｓ，２Ｈ），４．３７（ｑ，Ｊ＝７．２Ｈｚ，２Ｈ），３．０６（ｑ，Ｊ＝７．１Ｈｚ，４Ｈ），１．３７（ｔ，Ｊ＝７．１Ｈｚ，３Ｈ），０．９９（ｔ，Ｊ＝７．１Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１６３．５，１５７．４，１５６．２，１５５．２，１４８．６，１４４．０，１３５．８，１３２．１，１３２．１，１２９．８，１２８．９，１２５．０，１１９．２，１１３．３，１１１．７，１０６．７，６１．５，４５．６（２Ｃ），１４．２，１１．９（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ２２Ｈ２２Ｎ３Ｏ６（Ｍ−Ｈ）− ４２４．１５０９．Ｆｏｕｎｄ ４２４．１５１３；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ２２Ｈ２３Ｎ３Ｏ６：Ｃ，６２．１１；Ｈ，５．４５；Ｎ，９．８８．Ｆｏｕｎｄ：Ｃ，６２．０２；Ｈ，５．６５；Ｎ，９．７６．
<Synthesis of Compound 3l>
By a method similar to the synthesis of compound 3a, yellow powdery compound 3l was obtained from compound 1 and compound 21 in a yield of 62%.
Melting point: 187-188 ° C; 1 H NMR (500 MHz, CDCl 3 ) ... 8.44 (s, 1 H), 8.23 (d, J = 2.1 Hz, 1 H), 7.52 (dd, J = 8.6, 2.1 Hz, 1H), 7.31 (s, 1H), 6.89 (s, 1H), 6.86 (d, J = 8.6 Hz, 1H), 6.21 (br s, 2H), 4.37 (q, J = 7.2 Hz, 2H), 3.06 (q, J = 7.1 Hz, 4H), 1.37 (t, J = 7.1 Hz, 3H), 0.99 (t, J = 7.1 Hz, 6H); 13 C NMR (125 MHz, CDCl 3 ) 163.5, 157.4, 156.2, 155.2, 148.6, 144.0 135.8, 132.1, 132.1, 129.8, 128.9, 125.0, 119.2, 113.3, 111. , 106.7,61.5,45.6 (2C), 14.2,11.9 ( 2C); HRMS (ESI) calcd for C 22 H 22 N 3 O 6 (M-H) - 424.1509 . Found 424.1513; Anal. Calcd for C 22 H 23 N 3 O 6: C, 62.11; H, 5.45; N, 9.88. Found: C, 62.02; H, 5.65; N, 9.76.
＜化合物３ｍの合成＞
化合物３ｋ（１１０．４ｍｇ，０．２２１ｍｍｏｌ）をメタノール：テトラヒドロフラン（２：１）３ｍＬに溶かし、１０％Ｐｄ／Ｃ（パラジウム炭素、１７．０ｍｇ）を室温下で加え、水素雰囲気下で４０分間撹拌した。反応液をろ過後、カラムクロマトグラフィーにて精製し、黄色粉状物の化合物３ｍ（６６．０ｍｇ，７３％）を得た。
融点：２４９−２５０℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・１１．０６（ｂｒ ｓ，１Ｈ），８．４６（ｓ，１Ｈ），８．１５（ｄ，Ｊ＝８．３Ｈｚ，２Ｈ），７．５６（ｄ，Ｊ＝８．３Ｈｚ，２Ｈ），７．３３（ｓ，１Ｈ），６．８６（ｓ，１Ｈ），４．３６（ｑ，Ｊ＝７．１Ｈｚ，２Ｈ），３．０３（ｑ，Ｊ＝７．１Ｈｚ，４Ｈ），１．３６（ｔ，Ｊ＝７．１Ｈｚ，３Ｈ），０．９６（ｔ，Ｊ＝７．１Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１７０．９，１６３．６，１５７．４，１５６．７，１５４．９，１４８．６，１４６．１，１３２．７，１３０．８（２Ｃ），１３０．３，１２８．４（２Ｃ），１２８．１，１１３．４，１１１．４，１０６．２，６１．６，４６．０（２Ｃ），１４．３，１２．０（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ２３Ｈ２２ＮＯ６（Ｍ−Ｈ）− ４０８．１４４７．Ｆｏｕｎｄ ４０８．１４５０；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ２３Ｈ２３ＮＯ６・１／４Ｈ２Ｏ：Ｃ，６６．７４；Ｈ，５．７２；Ｎ，３．３８．Ｆｏｕｎｄ：Ｃ，６６．９８；Ｈ，５．７６；Ｎ，３．３５．
<Synthesis of Compound 3m>
Compound 3k (110.4 mg, 0.221 mmol) was dissolved in 3 mL of methanol: tetrahydrofuran (2: 1), 10% Pd / C (palladium carbon, 17.0 mg) was added at room temperature, and the mixture was stirred under a hydrogen atmosphere for 40 minutes. did. The reaction solution was filtered and then purified by column chromatography to obtain a yellow powdery compound 3m (66.0 mg, 73%).
Melting point: 249-250 ° C .; 1 H NMR (500 MHz, CDCl 3 )... 11.06 (br s, 1 H), 8.46 (s, 1 H), 8.15 (d, J = 8.3 Hz, 2H), 7.56 (d, J = 8.3 Hz, 2H), 7.33 (s, 1H), 6.86 (s, 1H), 4.36 (q, J = 7.1 Hz, 2H) , 3.03 (q, J = 7.1 Hz, 4H), 1.36 (t, J = 7.1 Hz, 3H), 0.96 (t, J = 7.1 Hz, 6H); 13 C NMR ( 125 MHz, CDCl 3 )... 170.9, 163.6, 157.4, 156.7, 154.9, 148.6, 146.1, 132.7, 130.8 (2C), 130.3 , 128.4 (2C), 128.1, 113.4, 111.4, 106.2, 61.6, 46.0 (2C). 14.3,12.0 (2C); HRMS (ESI ) calcd for C 23 H 22 NO 6 (M-H) - 408.1447. Found 408.1450; Anal. Calcd for C 23 H 23 NO 6 · 1 / 4H 2 O: C, 66.74; H, 5.72; N, 3.38. Found: C, 66.98; H, 5.76; N, 3.35.
＜化合物３ｎの合成＞
化合物３ｌ（１２０．９ｍｇ，０．２８４ｍｍｏｌ）を２．５ｍＬのメタノールに溶かし、１０％Ｐｄ／Ｃ（パラジウム炭素、３９．１ｍｇ）を室温下で加え、水素雰囲気下で２時間撹拌した。反応液をろ過後、カラムクロマトグラフィーにて精製し、黄色粉状物の化合物３ｎ（７２．５ｍｇ，６５％）を得た。
融点：１８５−１８６℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・８．４１（ｓ，１Ｈ），７．２３（ｓ，１Ｈ），６．８２−６．７５（ｍ，４Ｈ），４．３６（ｑ，Ｊ＝７．１Ｈｚ，２Ｈ），３．６８（ｂｒ ｓ，４Ｈ），３．０７（ｑ，Ｊ＝７．１Ｈｚ，４Ｈ），１．３７（ｔ，Ｊ＝７．１Ｈｚ，３Ｈ），０．９６（ｔ，Ｊ＝７．１Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１６３．９，１５７．９，１５６．１，１５５．３，１４９．０，１３４．７，１３４．０，１３２．５，１３２．４，１３１．５，１２０．０，１１６．９，１１６．４，１１２．０，１１０．８，１０５．２，６１．３，４５．５（２Ｃ），１４．２，１２．１（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ２２Ｈ２４Ｎ３Ｏ４（Ｍ−Ｈ）− ３９４．１７６７．Ｆｏｕｎｄ ３９４．１７６０；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ２２Ｈ２５Ｎ３Ｏ４・１／４Ｈ２Ｏ：Ｃ，６６．０７；Ｈ，６．４３；Ｎ，１０．５１．Ｆｏｕｎｄ：Ｃ，６６．０１；Ｈ，６．２７；Ｎ，１０．４４．
<Synthesis of Compound 3n>
Compound 31 (120.9 mg, 0.284 mmol) was dissolved in 2.5 mL of methanol, 10% Pd / C (palladium on carbon, 39.1 mg) was added at room temperature, and the mixture was stirred under a hydrogen atmosphere for 2 hours. The reaction solution was filtered and purified by column chromatography to obtain yellow powdery compound 3n (72.5 mg, 65%).
Melting point: 185-186 ° C; 1 H NMR (500 MHz, CDCl 3 ) ... 8.41 (s, 1 H), 7.23 (s, 1 H), 6.82-6.75 (m, 4 H), 4.36 (q, J = 7.1 Hz, 2H), 3.68 (br s, 4H), 3.07 (q, J = 7.1 Hz, 4H), 1.37 (t, J = 7. 1 Hz, 3H), 0.96 (t, J = 7.1 Hz, 6H); 13 C NMR (125 MHz, CDCl 3 )... 163.9, 157.9, 156.1, 155.3, 149. 0, 134.7, 134.0, 132.5, 132.4, 131.5, 120.0, 116.9, 116.4, 112.0, 110.8, 105.2, 61.3, 45.5 (2C), 14.2,12.1 (2C ); HRMS (ESI) calcd for C 22 24 N 3 O 4 (M- H) - 394.1767. Found 394. 1760; Anal. Calcd for C 22 H 25 N 3 O 4 · 1 / 4H 2 O: C, 66.07; H, 6.43; N, 10.51. Found: C, 66.01; H, 6.27; N, 10.44.
＜化合物３ｏの合成＞
化合物３ｎ（４７．８ｍｇ，０．１３１ｍｍｏｌ）をメタノール：水（１：４）２ｍＬに溶かし、酢酸（０．２ｍｌ）、亜硝酸ナトリウム（２１．４ｍｇ，０．３１０ｍｍｏｌ）を室温下で加えた。５分間撹拌後、反応液を酢酸エチルで希釈し、有機層を水、飽和食塩水で洗浄した。溶媒を留去後、カラムクロマトグラフィーにて精製し、黄色粉状物の化合物３ｏ（５０．２ｍｇ，９４％）を得た。
融点：２３２−２３３℃；１Ｈ ＮＭＲ（５００ＭＨｚ，ＣＤＣｌ３）・・・８．５１（ｓ，１Ｈ），８．０２（ｓ，１Ｈ），７．９８（ｄ，Ｊ＝８．６Ｈｚ，１Ｈ），７．５５（ｄｄ，Ｊ＝８．６，１．０Ｈｚ，１Ｈ），７．４１（ｓ，１Ｈ），６．９１（ｓ，１Ｈ），４．３８（ｑ，Ｊ＝７．１Ｈｚ，２Ｈ），３．０５（ｑ，Ｊ＝７．１Ｈｚ，４Ｈ），１．３６（ｔ，Ｊ＝７．１Ｈｚ，３Ｈ），０．９６（ｔ，Ｊ＝７．０Ｈｚ，６Ｈ）；１３Ｃ ＮＭＲ（１２５ＭＨｚ，ＣＤＣｌ３）・・・１６３．７，１５８．１，１５６．５，１５５．２，１４９．０，１３９．３，１３８．４，１３８．３，１３３．４，１３１．１，１２７．０，１１５．５，１１４．１，１１２．５，１１１．４，１０６．０，６１．７，５２．２，４６．０（２Ｃ），１４．３，１２．１（２Ｃ）；ＨＲＭＳ（ＥＳＩ）ｃａｌｃｄ ｆｏｒ Ｃ２２Ｈ２１Ｎ４Ｏ４（Ｍ−Ｈ）− ４０５．１５６３．Ｆｏｕｎｄ ４０５．１５６４；Ａｎａｌ．Ｃａｌｃｄ ｆｏｒ Ｃ２２Ｈ２２Ｎ４Ｏ４：Ｃ，６５．０１；Ｈ，５．４６；Ｎ，１３．７８．Ｆｏｕｎｄ：Ｃ，６４．７２；Ｈ，５．６０；Ｎ，１３．６８．
<Synthesis of Compound 3o>
Compound 3n (47.8 mg, 0.131 mmol) was dissolved in 2 mL of methanol: water (1: 4), and acetic acid (0.2 ml) and sodium nitrite (21.4 mg, 0.310 mmol) were added at room temperature. After stirring for 5 minutes, the reaction solution was diluted with ethyl acetate, and the organic layer was washed with water and saturated brine. After the solvent was distilled off, the residue was purified by column chromatography to obtain a yellow powdery compound 3o (50.2 mg, 94%).
Melting point: 232-233 ° C .; 1 H NMR (500 MHz, CDCl 3 )... 8.51 (s, 1 H), 8.02 (s, 1 H), 7.98 (d, J = 8.6 Hz, 1 H) ), 7.55 (dd, J = 8.6, 1.0 Hz, 1H), 7.41 (s, 1H), 6.91 (s, 1H), 4.38 (q, J = 7.1 Hz) , 2H), 3.05 (q, J = 7.1Hz, 4H), 1.36 (t, J = 7.1Hz, 3H), 0.96 (t, J = 7.0Hz, 6H); 13 C NMR (125 MHz, CDCl 3 ) ... 163.7, 158.1, 156.5, 155.2, 149.0, 139.3, 138.4, 138.3, 133.4, 131.1 , 127.0, 115.5, 114.1, 112.5, 111.4, 106.0, 61.7, 52.2, 46.0 (2C), 14.3,12.1 (2C) ; HRMS (ESI) calcd for C 22 H 21 N 4 O 4 (M-H) - 405.1563. Found 405.5644; Anal. Calcd for C 22 H 22 N 4 O 4: C, 65.01; H, 5.46; N, 13.78. Found: C, 64.72; H, 5.60; N, 13.68.
（実施例２：各化合物の機能性確認）
実施例２では、前記実施例１で合成した本発明の化合物（化合物３ａ〜３ｏ）の蛍光特性を検討し、各化合物が種々の機能性を有した蛍光物質であることを確認した。
(Example 2: Functional confirmation of each compound)
In Example 2, the fluorescence characteristics of the compounds of the present invention (compounds 3a to 3o) synthesized in Example 1 were examined, and it was confirmed that each compound was a fluorescent substance having various functions.
（１）蛍光センサー分子としての機能（特定の分子種を検出する機能）
水溶液（１ｍＭ リン酸バッファー（ｐＨ７．４））中での、各化合物３μＭの励起波長４２０ｎｍにおける蛍光スペクトルを、分光蛍光光度計（日本分光株式会社製、ＦＰ−６６００）を用いて測定した。
(1) Function as a fluorescent sensor molecule (function to detect specific molecular species)
The fluorescence spectrum of each compound 3 μM at an excitation wavelength of 420 nm in an aqueous solution (1 mM phosphate buffer (pH 7.4)) was measured using a spectrofluorometer (manufactured by JASCO Corporation, FP-6600).
結果の一部を図２に示す。化合物３ａ〜３ｏは、それぞれの構造の違いに応じて蛍光特性が異なり、したがって、本発明の化合物は例えば以下のような用途に好適であることが示された。
化合物３ｃから３ｂへの変換は、アミド基からアミノ基への変換であり、変換により全く無蛍光の状態になる（図２（ｉ））。こうした反応は、例えば生体内に存在するペプチダーゼの検出に適用することができる。
化合物３ｎから３ｏへの変換は、ジアミノ基からトリアゾールへの変換であり、変換により全くの無蛍光の状態から、蛍光を持つようになる（図２（ｉｉ））。こうした反応は、例えば生体内において血管収縮などを担う生理活性物質である一酸化窒素の検出に適用することができる。
化合物３ｇから３ｆへの変換は、メトキシ基から水酸基への変換であり、変換により蛍光強度が約５分の１に減少する（図２（ｉｉｉ））。こうした反応は、例えば生体内の脱アルキル化酵素やグリコシダーゼの検出に適用できると考えられる。
A part of the results is shown in FIG. The compounds 3a to 3o have different fluorescence characteristics depending on the difference in structure, and thus the compounds of the present invention have been shown to be suitable for the following uses, for example.
The conversion from compound 3c to 3b is a conversion from an amide group to an amino group, and the conversion results in a completely non-fluorescent state (FIG. 2 (i)). Such a reaction can be applied to, for example, detection of peptidases present in a living body.
The conversion from the compound 3n to 3o is conversion from a diamino group to triazole, and the fluorescence is obtained from the completely non-fluorescent state by the conversion (FIG. 2 (ii)). Such a reaction can be applied to, for example, detection of nitric oxide which is a physiologically active substance responsible for vasoconstriction in a living body.
The conversion from compound 3g to 3f is a conversion from a methoxy group to a hydroxyl group, and the fluorescence intensity is reduced to about 1/5 by the conversion (FIG. 2 (iii)). Such a reaction is considered to be applicable to, for example, detection of dealkylating enzymes and glycosidases in vivo.
（２）環境応答型蛍光分子としての機能（溶液の脂溶性の高さを検出する機能）
水溶液（１ｍＭ リン酸バッファー、ｐＨ７．４）中、メタノール中、アセトニトリル中、又は塩化メチレン中での各化合物３ａ〜３ｏの蛍光強度（ＦＩ）は、個々の化合物が最も強い蛍光強度を持つ励起波長、蛍光波長における蛍光強度を、分光蛍光光度計（日本分光株式会社製、ＦＰ−６６００）を用いて測定した。（なお、用いた溶媒の脂溶性の高さは、「塩化メチレン」＞「アセトニトリル」＞「メタノール」＞「水溶液」の順である。）
(2) Function as an environmentally responsive fluorescent molecule (function to detect the high lipid solubility of a solution)
The fluorescence intensity (FI) of each compound 3a to 3o in aqueous solution (1 mM phosphate buffer, pH 7.4), methanol, acetonitrile, or methylene chloride is the excitation wavelength at which each compound has the strongest fluorescence intensity. The fluorescence intensity at the fluorescence wavelength was measured using a spectrofluorometer (manufactured by JASCO Corporation, FP-6600). (In addition, the high lipophilicity of the solvent used is in the order of “methylene chloride”>“acetonitrile”>“methanol”> “aqueous solution”.)
結果を表１に示す。ほとんどの化合物が脂溶性の高い溶媒になるのに伴い蛍光強度が増大した。また、その変化量は個々の化合物で異なっていた。特に、化合物３ｆ及び化合物３ｈでは、水溶液中と塩化メチレン中で比較すると、２００倍以上の蛍光強度の増大が起こっていた。
これらの結果から、本発明の化合物が、溶液の脂溶性の高さを検出する用途に好適であることが示された。
The results are shown in Table 1. As most of the compounds became highly lipid-soluble solvents, the fluorescence intensity increased. The amount of change was different for each compound. In particular, in Compound 3f and Compound 3h, the fluorescence intensity increased by a factor of 200 or more when compared in aqueous solution and methylene chloride.
From these results, it was shown that the compound of this invention is suitable for the use which detects the high lipid solubility of a solution.
本発明の化合物は蛍光性を有する新規クマリン誘導体であり、その蛍光性を利用して種々の用途に好適に利用可能である。例えば、前記化合物は、イオン、小分子、酵素等の特定の分子種の検出や、溶液の脂溶性の高さの検出等に好適に利用可能である。 The compound of the present invention is a novel coumarin derivative having fluorescence, and can be suitably used for various applications by utilizing the fluorescence. For example, the compound can be suitably used for the detection of specific molecular species such as ions, small molecules, enzymes, etc., the detection of the lipid solubility of a solution, and the like.
Claims (6)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2007043878A JP2008208039A (en) | 2007-02-23 | 2007-02-23 | Coumarin derivatives and their uses |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2007043878A JP2008208039A (en) | 2007-02-23 | 2007-02-23 | Coumarin derivatives and their uses |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2008208039A true JP2008208039A (en) | 2008-09-11 |
Family
ID=39784670
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2007043878A Pending JP2008208039A (en) | 2007-02-23 | 2007-02-23 | Coumarin derivatives and their uses |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2008208039A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103435603A (en) * | 2013-08-22 | 2013-12-11 | 贵州大学 | Coumarin-oxacalix[3]arene fluorescent reagents as well as preparation method and application thereof |
| WO2020039708A1 (en) | 2018-08-23 | 2020-02-27 | 国立大学法人九州大学 | Organic electroluminescence element |
| CN117343201A (en) * | 2023-08-21 | 2024-01-05 | 中国科学院宁波材料技术与工程研究所 | A multifunctional cellulose-based fluorescent material and its preparation method and application |
-
2007
- 2007-02-23 JP JP2007043878A patent/JP2008208039A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103435603A (en) * | 2013-08-22 | 2013-12-11 | 贵州大学 | Coumarin-oxacalix[3]arene fluorescent reagents as well as preparation method and application thereof |
| WO2020039708A1 (en) | 2018-08-23 | 2020-02-27 | 国立大学法人九州大学 | Organic electroluminescence element |
| CN117343201A (en) * | 2023-08-21 | 2024-01-05 | 中国科学院宁波材料技术与工程研究所 | A multifunctional cellulose-based fluorescent material and its preparation method and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP3636653A1 (en) | Asymmetrical si rhodamine and rhodol synthesis | |
| Guo et al. | Rapid assembly of quinazolinone scaffold via copper-catalyzed tandem reaction of 2-bromobenzamides with aldehydes and aqueous ammonia: application to the synthesis of the alkaloid tryptanthrin | |
| CN104357044B (en) | A kind of fluorescent probe and its preparation method and application | |
| JP4899046B2 (en) | New luciferin derivatives | |
| US11319331B2 (en) | Probe for selective detection of hypochlorous acid (HOCl) under physiological condition, and related methods | |
| US8394850B2 (en) | Fluorescent probe specific to hydrogen peroxide | |
| Gu et al. | A novel self-calibrating strategy for real time monitoring of formaldehyde both in solution and solid phase | |
| WO1999051586A1 (en) | Reagent for singlet oxygen determination | |
| Wang et al. | Design and synthesis of an AIEgen with multiple functions: Solvatochromism, chromism, lipid droplet imaging | |
| Sun et al. | A sensitive and selective fluorescent probe for the detection of endogenous peroxynitrite (ONOO−) in living cells | |
| Kim et al. | Novel intramolecular π–π-interaction in a BODIPY system by oxidation of a single selenium center: Geometrical stamping and spectroscopic and spectrometric distinctions | |
| Tigreros et al. | Expeditious ethanol quantification present in hydrocarbons and distilled spirits: Extending photophysical usages of the pyrazolo [1, 5-a] pyrimidines | |
| Şenkuytu et al. | Bodipy decorated triazine chemosensors for Ag+ ions with high selectivity and sensitivity | |
| Ma et al. | Dihydropyridine-based fluorescence probe for nitric oxide | |
| JP2010215795A5 (en) | ||
| JP2008208039A (en) | Coumarin derivatives and their uses | |
| WO2021187531A1 (en) | Luminescent substrate compound | |
| CN106008510B (en) | Aggregation-induced luminescent fluorescent sensor for detecting Hg2+ and its preparation method and application | |
| CN111793070B (en) | [1,2,4] -triazole fused ring compounds with fluorescence characteristics and preparation method and application thereof | |
| Liu et al. | Viscosity probes towards different organelles with red emission based on an identical hemicyanine structure | |
| JP2012154693A (en) | Potassium fluorescent probe | |
| Ossowski et al. | Synthesis of lysine derivatives containing aza-crown ethers and a chromophore unit | |
| JP6360345B2 (en) | Xanthene compounds and uses thereof | |
| WO2017090631A1 (en) | Fluorescent probe for detecting extracellular metabolite and screening method employing said fluorescent probe | |
| JP4453528B2 (en) | New chemiluminescent reagent |