JPH04316573A - New compound cf-24, its use and production - Google Patents
New compound cf-24, its use and productionInfo
- Publication number
- JPH04316573A JPH04316573A JP11090891A JP11090891A JPH04316573A JP H04316573 A JPH04316573 A JP H04316573A JP 11090891 A JP11090891 A JP 11090891A JP 11090891 A JP11090891 A JP 11090891A JP H04316573 A JPH04316573 A JP H04316573A
- Authority
- JP
- Japan
- Prior art keywords
- compound
- culture
- strain
- properties
- saccharothrix
- 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
- 150000001875 compounds Chemical class 0.000 title claims abstract description 30
- 238000004519 manufacturing process Methods 0.000 title claims description 12
- 239000000126 substance Substances 0.000 claims abstract description 18
- 241000204098 Saccharothrix Species 0.000 claims abstract description 8
- 238000000034 method Methods 0.000 claims description 18
- 239000002246 antineoplastic agent Substances 0.000 claims description 7
- 239000004480 active ingredient Substances 0.000 claims description 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 abstract description 21
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 abstract description 12
- 230000000259 anti-tumor effect Effects 0.000 abstract description 10
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 abstract description 8
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 abstract description 6
- 238000012258 culturing Methods 0.000 abstract description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 abstract description 6
- 244000005700 microbiome Species 0.000 abstract description 5
- 241000082882 Saccharothrix sp. Species 0.000 abstract description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 2
- 239000001963 growth medium Substances 0.000 abstract description 2
- 238000001819 mass spectrum Methods 0.000 abstract description 2
- 238000010265 fast atom bombardment Methods 0.000 abstract 2
- 230000001580 bacterial effect Effects 0.000 description 9
- IAKHMKGGTNLKSZ-INIZCTEOSA-N (S)-colchicine Chemical compound C1([C@@H](NC(C)=O)CC2)=CC(=O)C(OC)=CC=C1C1=C2C=C(OC)C(OC)=C1OC IAKHMKGGTNLKSZ-INIZCTEOSA-N 0.000 description 8
- 229920001817 Agar Polymers 0.000 description 8
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 8
- 239000008272 agar Substances 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 8
- 239000002609 medium Substances 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- 238000000605 extraction Methods 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 5
- 230000001766 physiological effect Effects 0.000 description 5
- 239000000741 silica gel Substances 0.000 description 5
- 229910002027 silica gel Inorganic materials 0.000 description 5
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 4
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 4
- 229920002472 Starch Polymers 0.000 description 4
- 238000000862 absorption spectrum Methods 0.000 description 4
- 230000003115 biocidal effect Effects 0.000 description 4
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 description 4
- 229960001338 colchicine Drugs 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 239000000049 pigment Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 235000019698 starch Nutrition 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 3
- 235000002639 sodium chloride Nutrition 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- HYPYXGZDOYTYDR-HAJWAVTHSA-N 2-methyl-3-[(2e,6e,10e,14e)-3,7,11,15,19-pentamethylicosa-2,6,10,14,18-pentaenyl]naphthalene-1,4-dione Chemical compound C1=CC=C2C(=O)C(C/C=C(C)/CC/C=C(C)/CC/C=C(C)/CC/C=C(C)/CCC=C(C)C)=C(C)C(=O)C2=C1 HYPYXGZDOYTYDR-HAJWAVTHSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- RFSUNEUAIZKAJO-VRPWFDPXSA-N D-Fructose Natural products OC[C@H]1OC(O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-VRPWFDPXSA-N 0.000 description 2
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 239000003463 adsorbent Substances 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- 210000002421 cell wall Anatomy 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000007385 chemical modification Methods 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 239000012531 culture fluid Substances 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 235000013312 flour Nutrition 0.000 description 2
- 238000002523 gelfiltration Methods 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- FUKUFMFMCZIRNT-UHFFFAOYSA-N hydron;methanol;chloride Chemical compound Cl.OC FUKUFMFMCZIRNT-UHFFFAOYSA-N 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 150000002823 nitrates Chemical class 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 235000019198 oils Nutrition 0.000 description 2
- 150000003904 phospholipids Chemical class 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 235000020183 skimmed milk Nutrition 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 2
- 235000019143 vitamin K2 Nutrition 0.000 description 2
- 239000011728 vitamin K2 Substances 0.000 description 2
- 229940041603 vitamin k 3 Drugs 0.000 description 2
- GMKMEZVLHJARHF-UHFFFAOYSA-N (2R,6R)-form-2.6-Diaminoheptanedioic acid Natural products OC(=O)C(N)CCCC(N)C(O)=O GMKMEZVLHJARHF-UHFFFAOYSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- RZVAJINKPMORJF-UHFFFAOYSA-N Acetaminophen Chemical compound CC(=O)NC1=CC=C(O)C=C1 RZVAJINKPMORJF-UHFFFAOYSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- SRBFZHDQGSBBOR-HWQSCIPKSA-N L-arabinopyranose Chemical compound O[C@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-HWQSCIPKSA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- GXCLVBGFBYZDAG-UHFFFAOYSA-N N-[2-(1H-indol-3-yl)ethyl]-N-methylprop-2-en-1-amine Chemical compound CN(CCC1=CNC2=C1C=CC=C2)CC=C GXCLVBGFBYZDAG-UHFFFAOYSA-N 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- MUPFEKGTMRGPLJ-RMMQSMQOSA-N Raffinose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 MUPFEKGTMRGPLJ-RMMQSMQOSA-N 0.000 description 1
- 241000576755 Sclerotia Species 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- MUPFEKGTMRGPLJ-UHFFFAOYSA-N UNPD196149 Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC2C(C(O)C(O)C(CO)O2)O)O1 MUPFEKGTMRGPLJ-UHFFFAOYSA-N 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- PNNNRSAQSRJVSB-BXKVDMCESA-N aldehydo-L-rhamnose Chemical compound C[C@H](O)[C@H](O)[C@@H](O)[C@@H](O)C=O PNNNRSAQSRJVSB-BXKVDMCESA-N 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 239000002518 antifoaming agent Substances 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 235000012343 cottonseed oil Nutrition 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000001085 cytostatic effect Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000000921 elemental analysis Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- OAYLNYINCPYISS-UHFFFAOYSA-N ethyl acetate;hexane Chemical compound CCCCCC.CCOC(C)=O OAYLNYINCPYISS-UHFFFAOYSA-N 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000002143 fast-atom bombardment mass spectrum Methods 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000005187 foaming Methods 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 239000003120 macrolide antibiotic agent Substances 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- GMKMEZVLHJARHF-SYDPRGILSA-N meso-2,6-diaminopimelic acid Chemical compound [O-]C(=O)[C@@H]([NH3+])CCC[C@@H]([NH3+])C([O-])=O GMKMEZVLHJARHF-SYDPRGILSA-N 0.000 description 1
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000006916 nutrient agar Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000006877 oatmeal agar Substances 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 235000011009 potassium phosphates Nutrition 0.000 description 1
- 238000012746 preparative thin layer chromatography Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 229920002545 silicone oil Polymers 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000004317 sodium nitrate Substances 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- GRONZTPUWOOUFQ-UHFFFAOYSA-M sodium;methanol;hydroxide Chemical compound [OH-].[Na+].OC GRONZTPUWOOUFQ-UHFFFAOYSA-M 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 150000003445 sucroses Chemical class 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 230000004565 tumor cell growth Effects 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Pyrane Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
【発明の詳細な説明】
【0001】〔発明の背景〕
【産業上の利用分野】本発明は、新規物質に関し、更に
詳しくは抗腫瘍性を有する新規化合物CF−24に関す
る。
【0002】
【従来の技術】抗腫瘍物質に関しては、既に多数のもの
が医薬として実用化されている。一般に、化学物質の生
理活性はその化学構造に依存するところが大きいため、
抗腫瘍性を有する新規な化合物に対しては不断の希求が
あると言えよう。
【0003】〔発明の概要〕
【発明が解決しようとする課題】本発明は、上記の希求
に応えるものである。すなわち、本発明は、抗腫瘍性を
有する新規化合物を提供することを目的とするものであ
る。
【0004】
【課題を解決するための手段】本発明は、次式(I)で
示される化合物が抗腫瘍性を有することを見出すことに
より上記目的を達成した。すなわち、本発明による新規
化合物CF−24は次式(I)で示されるものである。
【化2】
本発明は、また、この物質の使用に関する。すなわち、
本発明による抗腫瘍剤は、上記式(I)で示される化合
物CF−24を有効成分として含むものである。本発明
は、更にまた、この物質の製造法に関する。すなわち、
本発明による上記式(I)で示されるCF−24の製造
法は、サッカロスリックス属に属しCF−24の生産能
を有する菌株を適当な培地で培養し、その培養物より化
合物CF−24を得ること、を特徴とするものである。
【0005】〔発明の具体的説明〕新規化合物CF−2
4
1)化学構造
本発明による新規化合物CF−24は、前記の式(I)
で示される化学構造を有する。この化学構造は、次のよ
うにして決定されたものである。CF−24のプロトン
核磁気共鳴スペクトル(図1)および炭素13核磁気共
鳴スペクトル(図2)の解析により、CF−24は、1
個の二重結合を有する14員環β−ケトマクロライド骨
格および3個のメチル基と3個の水酸基を有するアルキ
ル側鎖からなることが明らかとなった。スペクトルの更
に詳細な検討により、部分構造のつながりが、明らかと
なり、更に、マススペクトルより判明した分子量より、
CF−24の構造を式(I)のように決定した。
2)物理化学的性状
(1)外観:黄色油状
(2)溶解性:ベンゼン、クロロホルム、メタノール、
酢酸エチルに可溶。ヘキサン、水に不溶。
(3)Rf値(メルク社製「シリカゲル60F254
」使用):
クロロホルム−メタノール(19:1) 0.75ヘ
キサン−酢酸エチル(12:3) 0.71
(4)FAB−マススペクトル(m/z):(M+H)
+ 483
(5)元素分析:C 69.65%、H 10.4
5%、O 19.90%(計算値:C 69.67
%、H 10.44%,O 19.89%;C28
H50O6)(6)紫外吸収スペクトル λmax
nm(ε):図3に示す。
メタノール中:207(6370)
0.01規定塩酸−メタノール中:207(6370)
0.01規定水酸化ナトリウム−メタノール中:207
(20000 ),276(5450)(7)赤外吸収
スペクトル(クロロホルム中溶液法):図4に示す。
(8)プロトン核磁気共鳴スペクトル(500 メガヘ
ルツ、重クロロホルム中):図1に示す。
(9)炭素13核磁気共鳴スペクトル(125 メガヘ
ルツ、重クロロホルム中):図2に示す。
【0006】CF−24の製造
1)概要
化合物CF−24は現在のところ微生物の培養によって
のみ得られているが、類縁化合物の合成化学的修飾によ
って製造することも、あるいは全合成化学的に製造する
こともできよう。また、遺伝子工学的手法によることも
できよう。すなわち、化合物CF−24の産生に関与す
る遺伝子を適当な微生物に導入し、この様な形質転換微
生物を培養し、この培養物から得ることも可能であろう
。微生物の培養による場合の菌株としては、例えば、サ
ッカロスリックス属に属するCF−24生産能を有する
ものが使用される。具体的には、本発明者らの分離した
サッカロスリックスsp.2271−S3株がCF−2
4を生産することが本発明者らによって明らかにされて
いるが、その他の菌株については、抗生物質生産菌単離
の常法によって適当なものを自然界より分離することが
可能である。また、サッカロスリックスsp.2271
−S3株を含めてCF−24の生産菌を放射線照射その
他の変異処理に付して、CF−24の生産能を高める余
地も残されている。遺伝子工学的手法もまた可能である
ことは前記したところである
。2)2271−S3株
CF−24生産能を有するサッカロスリックス属の菌株
として本発明者らの見出している2271−S3株は、
下記の内容のものである。
(1)由来および寄託番号
2271−S3株は、埼玉県秩父郡横川町で採取した土
壌から分離されたものであり、平成3年4月10日に工
業技術院微生物工業技術研究所に寄託されて、「微工研
条寄第3350号」(FERM BP−3350)の
番号を得ている。
(2)菌学的性状
2271−S3株の菌学的性状は以下の通りである。な
お、本菌株の“特徴づけ”は特許庁の「産業別審査基準
」記載の方法によった。
■形態性状: 本菌株の基生菌糸は巾0.3〜0.8
μmで、分枝しながら長く伸長し、時には特徴的なジグ
ザグ状を呈し、短い桿状に分断する。気菌糸は主軸を長
く伸ばし、少くない分枝がレイス状に絡まり、全菌糸が
分節・分断して胞子鎖を形成する。胞子鎖は直状または
曲状で10から50以上の胞子から成り、成熟するとや
やジグザグ状を呈する。分節胞子は円筒形、巾0.3〜
0.5μm、長さ1.2〜1.7μm、平滑表面、非運
動性である。胞子のう、菌核、その他の特殊形態は観察
されなかった。
■化学的性状: 本菌株は全菌体アミノ酸としてメゾ
・ジアミノピメリン酸、全菌体糖パターンとしてガラク
トース、マンノース、ラムノースを含み、細胞壁化学型
はIII C+ラムノース型である。主要メナキノンは
MK−9(H4 )、リン脂質パターンはホスホチジル
エタノールアミンを含むPII型である。
■培養性状: 本菌株の培養性状は表1に示す。集落
の菌叢色は白色系列または灰色系列、裏面色は特徴のな
い不鮮明色または暗褐色、メラニン様色素・その他の拡
散性色素の産生は認められなかった。
■生理的性状: 本菌株の生理的性状は表2に示す。
本菌株は中温性で、利用診断糖はD−フラクトースを除
く全部を利用する。
3)分類的位置
以上の“特徴づけ”の結果に基づき、Bergey’s
Manual ofSystematic Bact
eriology, Vol. 4. により本菌株の
分類的位置を検索した。本菌株の属レベルの特徴を要約
すると、ノカルディオフォームの形態を示し、細胞壁化
学型はIII C+ラムノース型、主要メナキノンはM
K−9(H4 )、リン脂質パターンはPII型である
。これらの特徴をもつ属はSaccharothrix
のみである。よって、本菌株2271−S3はSac
charothrix 属に位置される。従って、本菌
株をSaccharothrix sp. 2271−
S3株と呼称する。
【0007】
表 1 培養性状
培 地 集落表面の
菌叢色 集落の裏面色
シュクロース・ 白色系列(a)
淡黄色(2ca〜2ea )
硝酸塩寒天
グルコース・アス
灰色系列(b〜c) 茶味灰色(2
ec〜2ig) パラギン寒天
グリセリ
ン・アス 灰色系列(b〜c)
茶味灰色(2ec〜2ig) パラギン寒天
後に暗褐灰色(2nl) 無機
塩・スターチ寒天 白色系列(a)
淡黄色(2ca) から
茶味白色(3ca)
チロシン寒天 白色系列
(a) 淡黄色(2ca〜3ea
)後に
淡
黄味褐色(2ic) 栄養寒天
気菌糸なし
淡黄色(2ca〜2db) イー
スト・麦芽寒天 灰色系列(c〜f 〜10fe
) 淡黄色(2ca)から淡黄味
橙色(3ea) ,後に暗褐色
オートミール寒天 灰色系列(c〜b)
淡黄色(2ca〜2ea)
・培地中への拡散性色素の産生は認められなかっ
た。
・( )内はカラー・ハーモニー・マニュアル(
コンテナー・コーポレーショ ン・オブ・アメリ
カ、1950)の色標コード。
【0008】
表 2 生理的性状
生育温度範囲
25〜37℃
最適温度
27〜30℃ メラニン
様色素
チロシン寒天
−
ペプトン・イースト鉄寒天
− トリプトン・イースト・ブ
ロス − スターチ
の加水分解
+ ゼラチンの液化
+
脱脂粉乳のペプトン化
+ 脱脂粉乳の凝固
− 硝酸塩の還元
+
炭素源の同化
D−グルコース
+
L−アラビノース
+ D−キシロース
+
D−フラクトース
− シ
ュクロース
+ L−ラムノース
+ ラフィノース
+
i−イノシトール
+ D−マンニット
+
【0009】培養/CF−24の生産
化合物CF−24は、サッカロスリックス属に属するC
F−24生産菌を適当な培地で好気的に培養し、その培
養物から目的物を採取することにより製造することがで
きる。培地は、CF−24生産菌が利用し得る任意の栄
養源を含有するもので有り得る。具体的には、例えば炭
素源としてグルコース、シュークロース、マルトース、
スターチおよび油脂類などが使用でき、窒素源として大
豆粉、綿実粕、乾燥酵母、酵母エキスおよびコーンステ
ィープリカーなどの有機物並びにアンモニウム塩または
硝酸塩、例えば、硫酸アンモニウム、硝酸ナトリウムお
よび塩化アンモニウムなどの無機物が利用できる。また
、必要に応じて、塩化ナトリウム、塩化カリウム、燐酸
塩、重金属塩など無機塩類を添加することができる。
発酵中の発泡を抑制するために、常法に従って適当な消
泡剤、例えばシリコーン油を添加することもできる。培
養方法としては、一般に行なわれている抗生物質の生産
方法と同じく、好気的液体培地培養法が最も適している
。培養温度は、20〜37℃が適当であるが、25〜3
0℃が好ましい。この方法でCF−24の生産量は、振
盪培養、通気攪拌培養ともに通常、培養3〜4日間で最
高に達する。このようにして、CF−24の蓄積された
培養物が得られる。培養物中では、CF−24はその一
部は培養槇液中に存在するが、その大部分は菌体中に存
在する。このような培養物からCF−24を採取するに
は、合目的的な任意の方法が利用可能である。その一つ
の方法は、抽出の原理に基づくものであって、具体的に
は、培養槇液中のCF−24についてはこれを水不混和
性のCF−24用溶媒(前記CF−24の物理化学的性
状の項参照)、例えば酢酸エチルなどで抽出する方法、
あるいは菌体内のCF−24については槇過、遠心分離
などで得た菌体集体をメタノール、エタノール、アセト
ンなどで処理して回収する方法などがある。菌体を分離
せずに培養物そのままを上記の抽出操作に付すこともで
きる。適当な溶媒を用いた向流分配法も抽出の範疇に入
れることができる。培養物からCF−24を採取する他
の一つの方法は、吸着の原理に基づくものであって、既
に液状となっているCF−24含有物、例えば培養槇液
あるいは上記のようにして抽出操作を行なうことにより
得られる抽出液を対象として、適当な吸着剤、例えばシ
リカゲル、活性炭、「ダイヤイオンHP−20」(三菱
化成社製)などを用いて目的のCF−24を吸着させ、
その後、適当な溶媒にて溶離させることによってCF−
24を得ることができる。このようにして得られたCF
−24溶液を減圧濃縮乾固すれば、CF−24粗標品が
得られる。このようにして得られたCF−24の粗標品
を更に精製するためには、上記の抽出法および吸着法に
ゲル槇過法、高速液体クロマトグラフィーなどを必要に
応じて組合せて必要回数行なえばよい。例えば、シリカ
ゲルなどの吸着剤、「セファデックスLH−20」(フ
ァルマシア社製)などのゲル槇過剤を用いたカラムクロ
マトグラフィー、「YMCパック」(山村科学社製)な
どを用いた高速液体クロマトグラフィーおよび向流分配
法を適宜組合せて実施することができる。具体的には、
例えば、CF−24粗標品を「セファデックスLH−2
0」カラムに付し、クロロホルム−メタノール(1:1
)混合液で活性画分を溶出させ、濃縮乾固するとCF−
24の純品が得られる。
【0010】CF−24の用途
本発明による化合物CF−24は、抗腫瘍活性を有する
という点で有用である。
(1)生物活性
CF−24は腫瘍細胞に対して細胞増殖抑制活性を示し
た。例えば、ヒトの腫瘍細胞であるKB細胞またはコル
ヒチン耐性KB細胞を1×105 /mlとなるように
10%熱非働化牛胎児血清を含むMEM培地に調製し、
種々の濃度のCF−24とともに37℃で、24時間培
養した後のIC50値は、それぞれ6.0μg/mlで
あった。
また、コルヒチン耐性KB細胞に対するコルヒチン存在
下でのCF−24の細胞増殖抑制活性を上記と同様に測
定した時のIC50値は1.0μg/ml以下であった
。このことからCF−24は、単独においても、また、
コルヒチンと相乗的に作用してより強い腫瘍細胞増殖抑
制を示すことが明らかとなった。上記のように、本発明
のCF−24は抗腫瘍活性を示すことが明らかにされた
。従って、本発明のCF−24は抗腫瘍剤として使用す
ることができる。
(2)抗腫瘍剤
このように、本発明によるCF−24はヒトの腫瘍、特
に悪性腫瘍に対して抗腫瘍性を示すことが明らかにされ
た。従って、本発明化合物は、抗腫瘍剤もしくは腫瘍治
療剤として使用することができる。抗腫瘍剤としての本
発明化合物は、合目的的な任意の投与経路で、また、採
用投与経路によって決る剤型で投与することができる。
薬剤としては、製薬上許容される担体あるいは希釈剤で
希釈された形態が普通である。抗腫瘍剤としての本発明
化合物を実際に投与する場合には、これらを製薬上許容
される溶剤に溶解して注射する方法が代表的なものの一
つとして挙げられる。具体的には、腹腔内注射、皮下注
射、静脈または動脈への血管内注射および注射による局
所投与などの方法がある。本発明化合物の投与量は、動
物試験の結果および種々の状況を勘案して、連続的また
は間欠的に投与した時に総投与量が一定量を越えないよ
うに定められる。具体的な投与量は、投与方法、患者ま
たは被処理動物の状況、例えば年齢、体重、性別、感受
性、食餌、投与時間、併用する薬剤、患者またはその病
気の程度に応じて変化することは言うまでも無く、また
、一定の条件のもとにおける適量と投与回数は、上記指
針を基にして専門医の適量決定試験によって決定されな
ければならない。具体的には、成人1日当り0.01〜
1g程度である。
【0011】
【発明の効果】本発明による新規化合物CF−24は、
優れた抗腫瘍作用を有している。本発明化合物がこのよ
うな生理的作用を有しているという特性は当業者にとっ
て思いがけなかったことと解される。
【0012】〔実験例〕
【実施例】以下の実験例は、本発明を更に詳しく説明す
るためのものであり、これによって本発明は限定される
ものではない。
(1)培養
使用した培地は、下記の組成の成分を1リットルの水に
溶解して、pH6.2に調整したものである。
スターチ 25g
大豆粉 15g乾燥酵母
2g炭酸カルシウム 4
g
50リットル容ジャーファーメンターに上記培地30リ
ットルを調製、殺菌したものへ、サッカロスリックスs
p.2271−S3株を接種し、27℃にて3日間、通
気攪拌培養を行なった。
(2)CF−24の採取
上記の条件で培養後、培養液(30リットル)を遠心分
離し、菌体を5リットルのアセトンで抽出した。抽出液
を濃縮後、500mlの酢酸エチルで3回抽出し、濃縮
乾固した。これをシリカゲル(和光純薬社製「ワコーゲ
ルC200」)のカラムに吸着させ、クロロホルム−メ
タノール(150:1)混合液で溶出した。活性フラク
ションを濃縮乾固した後、これを調製用薄層クロマトグ
ラフィー(メルク社製「シリカゲル60F254 TL
Cプレート」使用)に付した。クロロホルム−メタノー
ル(19:1)混合液で展開した後、活性画分をかき取
り、これより、メタノールでCF−24を抽出した。抽
出液を逆相SEP−PAK(ウォーターズ・アソシエー
ツ社製「C18カートリッジ」)に付し、メタノール−
水(87:13)混合液で溶出した後、濃縮乾固してC
F−24の純品5mgを得た。Description: BACKGROUND OF THE INVENTION [Field of Industrial Application] The present invention relates to a novel substance, and more particularly to a novel compound CF-24 having antitumor properties. [0002] Regarding antitumor substances, a large number of them have already been put into practical use as medicines. Generally, the physiological activity of a chemical substance largely depends on its chemical structure;
It can be said that there is a constant desire for new compounds with antitumor properties. [Summary of the Invention] [Problem to be Solved by the Invention] The present invention satisfies the above-mentioned desire. That is, an object of the present invention is to provide a novel compound having antitumor properties. Means for Solving the Problems The present invention has achieved the above object by discovering that a compound represented by the following formula (I) has antitumor properties. That is, the novel compound CF-24 according to the present invention is represented by the following formula (I). embedded image The invention also relates to the use of this substance. That is,
The antitumor agent according to the present invention contains the compound CF-24 represented by the above formula (I) as an active ingredient. The invention furthermore relates to a method for producing this material. That is,
The method for producing CF-24 represented by the above formula (I) according to the present invention involves culturing a strain belonging to the genus Saccharothrix and having the ability to produce CF-24 in an appropriate medium, and producing the compound CF-24 from the culture. It is characterized by obtaining. [Specific description of the invention] New compound CF-2
4 1) Chemical structure The novel compound CF-24 according to the present invention has the above formula (I)
It has the chemical structure shown below. This chemical structure was determined as follows. Analysis of the proton nuclear magnetic resonance spectrum (Figure 1) and carbon-13 nuclear magnetic resonance spectrum (Figure 2) of CF-24 revealed that CF-24 has 1
It was revealed that it consists of a 14-membered β-keto macrolide skeleton having 3 double bonds and an alkyl side chain having 3 methyl groups and 3 hydroxyl groups. A more detailed examination of the spectra revealed the relationship between the substructures, and from the molecular weight determined from the mass spectrum,
The structure of CF-24 was determined as shown in formula (I). 2) Physicochemical properties (1) Appearance: yellow oil (2) Solubility: benzene, chloroform, methanol,
Soluble in ethyl acetate. Insoluble in hexane and water. (3) Rf value (Silica gel 60F254 manufactured by Merck & Co., Ltd.
” used): Chloroform-methanol (19:1) 0.75 Hexane-ethyl acetate (12:3) 0.71
(4) FAB-mass spectrum (m/z): (M+H)
+ 483 (5) Elemental analysis: C 69.65%, H 10.4
5%, O 19.90% (calculated value: C 69.67
%, H 10.44%, O 19.89%; C28
H50O6) (6) Ultraviolet absorption spectrum λmax
nm (ε): Shown in FIG. In methanol: 207 (6370) In 0.01N hydrochloric acid-methanol: 207 (6370)
0.01N sodium hydroxide in methanol: 207
(20000), 276 (5450) (7) Infrared absorption spectrum (solution method in chloroform): Shown in FIG. (8) Proton nuclear magnetic resonance spectrum (500 MHz, in deuterated chloroform): Shown in FIG. (9) Carbon-13 nuclear magnetic resonance spectrum (125 MHz, in deuterated chloroform): Shown in FIG. Production of CF-24 1) Overview Compound CF-24 is currently obtained only by culturing microorganisms, but it can also be produced by synthetic chemical modification of related compounds or by total synthetic chemical modification. You could also do that. It may also be possible to use genetic engineering techniques. That is, it would be possible to introduce a gene involved in the production of compound CF-24 into a suitable microorganism, culture such a transformed microorganism, and obtain the compound from this culture. When culturing a microorganism, for example, a strain belonging to the genus Saccharothrix and capable of producing CF-24 is used. Specifically, the present inventors' isolated Saccharothrix sp. 2271-S3 strain is CF-2
Although the present inventors have shown that antibiotic-producing strains produce antibiotic-producing bacteria, it is possible to isolate suitable strains from nature using conventional methods for isolating antibiotic-producing bacteria. In addition, Saccharothrix sp. 2271
There is also room to increase CF-24 production ability by subjecting CF-24-producing bacteria, including the -S3 strain, to irradiation or other mutation treatments. As mentioned above, genetic engineering techniques are also possible. 2) Strain 2271-S3 The strain 2271-S3, which the present inventors have discovered as a strain of the genus Saccharothrix that has the ability to produce CF-24, is
The contents are as follows. (1) Origin and deposit number Strain 2271-S3 was isolated from soil collected in Yokokawa-cho, Chichibu-gun, Saitama Prefecture, and was deposited at the Institute of Microbial Technology, Agency of Industrial Science and Technology on April 10, 1991. As a result, we obtained the number ``FERM BP-3350'' (FERM BP-3350). (2) Mycological properties The mycological properties of strain 2271-S3 are as follows. The "characterization" of this strain was carried out in accordance with the method described in the "Industrial Examination Standards" of the Japan Patent Office. ■Morphological characteristics: The basal hyphae of this strain have a width of 0.3 to 0.8
μm long, branching and elongating, sometimes exhibiting a characteristic zigzag shape, and dividing into short rod shapes. Aerial hyphae elongate their main axis, and a few branches intertwine in a lacy pattern, and all hyphae segment and divide to form spore chains. The spore chain is straight or curved and consists of 10 to 50 or more spores, and when mature, it takes on a slightly zigzag shape. Segmental spores are cylindrical, width 0.3~
0.5 μm, length 1.2-1.7 μm, smooth surface, non-motile. No sporangia, sclerotia, or other special forms were observed. ■Chemical properties: This strain contains meso-diaminopimelic acid as a total bacterial amino acid, galactose, mannose, and rhamnose as a total bacterial sugar pattern, and the cell wall chemical type is III C+rhamnose type. The major menaquinone is MK-9 (H4), and the phospholipid pattern is type PII, including phosphotidylethanolamine. ■Culture properties: The culture properties of this strain are shown in Table 1. The bacterial flora color of the colony was white or gray, the color of the underside was indistinct or dark brown, and no production of melanin-like pigments or other diffusible pigments was observed. ■Physiological properties: The physiological properties of this strain are shown in Table 2. This strain is mesophilic and uses all diagnostic sugars except D-fructose. 3) Based on the results of "characterization" beyond categorical position, Bergey's
Manual of Systematic Bact
eriology, Vol. 4. We searched for the taxonomic position of this strain. To summarize the genus-level characteristics of this strain, it shows a nocardioform morphology, the cell wall chemotype is III C + rhamnose type, and the main menaquinone is M
K-9 (H4), phospholipid pattern is PII type. The genus with these characteristics is Saccharothrix
Only. Therefore, this strain 2271-S3 is Sac
It is placed in the genus Charothrix. Therefore, this strain was classified as Saccharothrix sp. 2271-
It is called the S3 strain. Table 1 Culture properties Medium Bacterial flora color on the surface of the colony Color on the back side of the colony
Sucrose/white series (a)
Pale yellow (2ca~2ea)
nitrate agar
glucose as
Gray series (b to c) Brownish gray (2
ec~2ig) paragine agar
Glycerin As Gray series (b-c)
Brown gray (2ec~2ig) Paragin agar
Later dark brownish gray (2nl) Inorganic salt/starch agar White series (a)
From pale yellow (2ca)
Brownish white (3ca)
Tyrosine agar White series (a) Pale yellow (2ca~3ea
)later
Light yellowish brown (2ic) Nutrient agar
No aerial mycelium
Pale yellow (2ca~2db) Yeast/malt agar Gray series (c~f~10fe
) Pale yellow (2ca) to light yellowish
Orange (3ea), later dark brown Oatmeal agar Gray series (c-b)
Pale yellow (2ca-2ea)
・Production of diffusible pigment into the medium was not observed.・The information in parentheses is the Color Harmony Manual (
Container Corporation of America, 1950) color standard code. Table 2 Physiological properties Growth temperature range
25-37℃
optimal temperature
27-30℃ Melanin-like pigment Tyrosine agar
−
Peptone yeast iron agar
− Tryptone yeast broth − Hydrolysis of starch
+ Liquefaction of gelatin
+
Peptonization of skim milk powder
+ Coagulation of skim milk powder
- reduction of nitrates;
+
Carbon source assimilation D-glucose
+
L-arabinose
+ D-xylose
+
D-fructose
- sucrose
+ L-rhamnose
+ Raffinose
+
i-inositol
+ D-Mannit
+Culture/Production of CF-24 The compound CF-24 is derived from C belonging to the genus Saccharothrix.
It can be produced by culturing F-24 producing bacteria aerobically in an appropriate medium and collecting the target product from the culture. The medium can contain any nutrient source that can be utilized by the CF-24 producing bacteria. Specifically, for example, glucose, sucrose, maltose,
Starches and fats and oils can be used as nitrogen sources, as well as organic substances such as soy flour, cottonseed meal, dried yeast, yeast extract and corn steep liquor, and inorganic substances such as ammonium salts or nitrates, such as ammonium sulfate, sodium nitrate and ammonium chloride. Available. Moreover, inorganic salts such as sodium chloride, potassium chloride, phosphates, and heavy metal salts can be added as necessary. In order to suppress foaming during fermentation, suitable antifoaming agents such as silicone oil can also be added according to conventional methods. The most suitable culture method is the aerobic liquid culture method, which is the same as the commonly used antibiotic production method. The appropriate culture temperature is 20-37°C, but 25-37°C is suitable.
0°C is preferred. In this method, the production amount of CF-24 usually reaches its maximum within 3 to 4 days of culture in both shaking culture and aerated agitation culture. In this way, a culture enriched with CF-24 is obtained. In the culture, a part of CF-24 exists in the culture fluid, but most of it exists in the bacterial cells. Any convenient method can be used to harvest CF-24 from such cultures. One method is based on the principle of extraction, and specifically, for CF-24 in the culture fluid, it is extracted with a water-immiscible CF-24 solvent (the physical (See the section on chemical properties), for example, extraction with ethyl acetate, etc.
Alternatively, for CF-24 in bacterial cells, there is a method in which a bacterial mass obtained by filtration, centrifugation, etc. is treated with methanol, ethanol, acetone, etc. and recovered. The culture itself can also be subjected to the above extraction operation without separating the bacterial cells. Countercurrent distribution methods using suitable solvents can also be included in the category of extraction. Another method for collecting CF-24 from a culture is based on the principle of adsorption, in which the CF-24-containing material is already in liquid form, such as the culture broth, or the extraction operation is performed as described above. The target CF-24 is adsorbed using an appropriate adsorbent, such as silica gel, activated carbon, "Diaion HP-20" (manufactured by Mitsubishi Chemical Corporation), etc., on the extract obtained by performing
Thereafter, CF-
24 can be obtained. CF obtained in this way
-24 solution is concentrated to dryness under reduced pressure to obtain a crude sample of CF-24. In order to further purify the crude sample of CF-24 obtained in this way, the extraction method and adsorption method described above may be combined with gel filtration method, high performance liquid chromatography, etc. as necessary, and the procedure may be performed as many times as necessary. Bye. For example, column chromatography using adsorbents such as silica gel, gel filtration agents such as "Sephadex LH-20" (manufactured by Pharmacia), and high-performance liquid chromatography using "YMC Pack" (manufactured by Yamamura Kagaku), etc. It is possible to carry out appropriate combinations of graphography and countercurrent distribution methods. in particular,
For example, a crude sample of CF-24 is
0” column and chloroform-methanol (1:1
) The active fraction was eluted with a mixture and concentrated to dryness, resulting in CF-
24 pure products are obtained. Uses of CF-24 Compound CF-24 according to the present invention is useful in that it has antitumor activity. (1) Biologically active CF-24 showed cytostatic activity against tumor cells. For example, human tumor cells such as KB cells or colchicine-resistant KB cells are prepared in a MEM medium containing 10% heat-inactivated fetal bovine serum to a concentration of 1 x 10 cells/ml,
The IC50 values after 24 hours of incubation at 37°C with various concentrations of CF-24 were 6.0 μg/ml, respectively. Furthermore, when the cell proliferation inhibitory activity of CF-24 in the presence of colchicine against colchicine-resistant KB cells was measured in the same manner as above, the IC50 value was 1.0 μg/ml or less. From this, CF-24 can be used alone as well.
It was revealed that it acts synergistically with colchicine to exhibit stronger inhibition of tumor cell growth. As mentioned above, it was revealed that the CF-24 of the present invention exhibits antitumor activity. Therefore, CF-24 of the present invention can be used as an antitumor agent. (2) Antitumor agent It has thus been revealed that CF-24 according to the present invention exhibits antitumor properties against human tumors, particularly malignant tumors. Therefore, the compounds of the present invention can be used as antitumor agents or tumor therapeutic agents. The compounds of the present invention as antitumor agents can be administered by any convenient route of administration and in a dosage form determined by the route of administration employed. The drug is usually in a diluted form with a pharmaceutically acceptable carrier or diluent. When actually administering the compounds of the present invention as antitumor agents, one typical method is to dissolve them in a pharmaceutically acceptable solvent and inject them. Specifically, methods include intraperitoneal injection, subcutaneous injection, intravascular injection into a vein or artery, and local administration by injection. The dosage of the compound of the present invention is determined in consideration of the results of animal tests and various situations so that the total dosage does not exceed a certain amount when administered continuously or intermittently. The specific dosage will vary depending on the method of administration, the circumstances of the patient or treated animal, such as age, weight, sex, sensitivity, diet, time of administration, concomitant drugs, and the severity of the patient or his or her illness. Needless to say, the appropriate dose and frequency of administration under certain conditions must be determined by a specialist's appropriate dose determination test based on the above guidelines. Specifically, 0.01~ per day for adults
It is about 1g. Effects of the Invention The novel compound CF-24 according to the present invention has the following properties:
It has excellent antitumor effects. It is understood that the property that the compounds of the present invention have such physiological effects was unexpected for those skilled in the art. [Experimental Examples] [Example] The following experimental examples are intended to explain the present invention in more detail, and the present invention is not limited thereby. (1) The culture medium used was one in which the following components were dissolved in 1 liter of water and the pH was adjusted to 6.2. Starch 25g Soy flour 15g Dry yeast
2g calcium carbonate 4
g Prepare 30 liters of the above medium in a 50 liter jar fermenter, sterilize it, and add Saccharothrix s.
p. 2271-S3 strain was inoculated and cultured with aeration at 27°C for 3 days. (2) Collection of CF-24 After culturing under the above conditions, the culture solution (30 liters) was centrifuged, and the bacterial cells were extracted with 5 liters of acetone. The extract was concentrated, extracted three times with 500 ml of ethyl acetate, and concentrated to dryness. This was adsorbed onto a column of silica gel (Wako Gel C200, manufactured by Wako Pure Chemical Industries, Ltd.), and eluted with a chloroform-methanol (150:1) mixture. After concentrating the active fraction to dryness, it was subjected to preparative thin layer chromatography (Silica Gel 60F254 TL manufactured by Merck & Co., Ltd.).
C plate). After developing with a chloroform-methanol (19:1) mixture, the active fraction was scraped off, from which CF-24 was extracted with methanol. The extract was subjected to reverse-phase SEP-PAK (“C18 cartridge” manufactured by Waters Associates), and methanol-
After elution with a mixture of water (87:13), it was concentrated to dryness and the C
5 mg of pure F-24 was obtained.
【図1】CF−24の重クロロホルム中における500
メガヘルツプロトン核磁気共鳴スペクトルを模写したも
のである。Figure 1: CF-24 at 500% in deuterated chloroform
This is a copy of the megahertz proton nuclear magnetic resonance spectrum.
【図2】CF−24の重クロロホルム中における125
メガヘルツ炭素13核磁気共鳴スペクトルを模写したも
のである。Figure 2: 125 of CF-24 in deuterated chloroform
This is a reproduction of a megahertz carbon-13 nuclear magnetic resonance spectrum.
【図3】CF−24のメタノール中(実線)、0.01
規定塩酸−メタノール中(実線)及び0.01規定水酸
化ナトリウム−メタノール中(点線)での紫外吸収スペ
クトルを模写したものである。[Figure 3] CF-24 in methanol (solid line), 0.01
This is a reproduction of the ultraviolet absorption spectra in normal hydrochloric acid-methanol (solid line) and in 0.01N sodium hydroxide-methanol (dotted line).
【図4】CF−24のクロロホルム中溶液法による赤外
吸収スペクトルを模写したものである。FIG. 4 is a reproduction of the infrared absorption spectrum of CF-24 obtained by a solution method in chloroform.
Claims (3)
成分として含む抗腫瘍剤。2. An antitumor agent comprising the compound CF-24 according to claim 1 as an active ingredient.
生産能を有する菌株を適当な培地で培養し、その培養物
より化合物CF−24を得ることを特徴とする、請求項
1に記載の化合物CF−24の製造法。3. The method according to claim 1, wherein a strain belonging to the genus Saccharothrix and capable of producing CF-24 is cultured in an appropriate medium, and the compound CF-24 is obtained from the culture. Method for producing compound CF-24.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP11090891A JPH04316573A (en) | 1991-04-16 | 1991-04-16 | New compound cf-24, its use and production |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP11090891A JPH04316573A (en) | 1991-04-16 | 1991-04-16 | New compound cf-24, its use and production |
Publications (1)
Publication Number | Publication Date |
---|---|
JPH04316573A true JPH04316573A (en) | 1992-11-06 |
Family
ID=14547703
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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JP11090891A Pending JPH04316573A (en) | 1991-04-16 | 1991-04-16 | New compound cf-24, its use and production |
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Country | Link |
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JP (1) | JPH04316573A (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2008500035A (en) * | 2004-05-26 | 2008-01-10 | アンスティテュ ナシオナル ポリテクニク ドゥ トゥールーズ | Novel saccharolytic strains and antibiotics derived from them, namely mutactomycin and aldogomycin |
-
1991
- 1991-04-16 JP JP11090891A patent/JPH04316573A/en active Pending
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2008500035A (en) * | 2004-05-26 | 2008-01-10 | アンスティテュ ナシオナル ポリテクニク ドゥ トゥールーズ | Novel saccharolytic strains and antibiotics derived from them, namely mutactomycin and aldogomycin |
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