JP3943082B2 - Substance having molecular structure of 8- (c-β-D-glucopyranosyl) -7,3 ', 4'-trihydroxyflavone, separation method thereof and drug composition using the same - Google Patents
Substance having molecular structure of 8- (c-β-D-glucopyranosyl) -7,3 ', 4'-trihydroxyflavone, separation method thereof and drug composition using the same Download PDFInfo
- Publication number
- JP3943082B2 JP3943082B2 JP2003580351A JP2003580351A JP3943082B2 JP 3943082 B2 JP3943082 B2 JP 3943082B2 JP 2003580351 A JP2003580351 A JP 2003580351A JP 2003580351 A JP2003580351 A JP 2003580351A JP 3943082 B2 JP3943082 B2 JP 3943082B2
- Authority
- JP
- Japan
- Prior art keywords
- glucopyranosyl
- trihydroxyflavone
- separation method
- extract
- trihydroxy
- 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.)
- Expired - Fee Related
Links
- 239000003814 drug Substances 0.000 title claims description 11
- 229940079593 drug Drugs 0.000 title claims description 11
- 238000000926 separation method Methods 0.000 title claims description 9
- 239000000203 mixture Substances 0.000 title claims description 8
- 239000000126 substance Substances 0.000 title claims description 8
- OCFBBQMGLIJSJY-UHFFFAOYSA-N vijayoside Natural products OC1C(O)C(O)C(CO)OC1C1=C(O)C=CC2=C1OC(C=1C=C(O)C(O)=CC=1)=CC2=O OCFBBQMGLIJSJY-UHFFFAOYSA-N 0.000 title description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 30
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 27
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 22
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical group CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 21
- 239000000284 extract Substances 0.000 claims description 21
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 11
- 238000003820 Medium-pressure liquid chromatography Methods 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 claims description 9
- 241000196324 Embryophyta Species 0.000 claims description 8
- 239000002904 solvent Substances 0.000 claims description 8
- 238000003818 flash chromatography Methods 0.000 claims description 7
- 239000003960 organic solvent Substances 0.000 claims description 7
- 235000001674 Agaricus brunnescens Nutrition 0.000 claims description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 6
- 240000008976 Pterocarpus marsupium Species 0.000 claims description 5
- 235000010453 Pterocarpus marsupium Nutrition 0.000 claims description 5
- 206010012601 diabetes mellitus Diseases 0.000 claims description 5
- 239000002798 polar solvent Substances 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 5
- 239000003586 protic polar solvent Substances 0.000 claims description 5
- 230000037396 body weight Effects 0.000 claims description 4
- 239000012141 concentrate Substances 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 3
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 3
- 239000003208 petroleum Substances 0.000 claims description 3
- 239000008194 pharmaceutical composition Substances 0.000 claims description 3
- 238000000227 grinding Methods 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims 1
- 239000011259 mixed solution Substances 0.000 claims 1
- 239000002994 raw material Substances 0.000 claims 1
- 150000001875 compounds Chemical class 0.000 description 12
- 241000218691 Cupressaceae Species 0.000 description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 7
- 239000000741 silica gel Substances 0.000 description 7
- 229910002027 silica gel Inorganic materials 0.000 description 7
- 239000006286 aqueous extract Substances 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000002024 ethyl acetate extract Substances 0.000 description 4
- 230000002218 hypoglycaemic effect Effects 0.000 description 4
- PFTAWBLQPZVEMU-DZGCQCFKSA-N (+)-3',4',5,7-Tetrahydroxy-2,3-trans-flavan-3-ol Natural products C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-DZGCQCFKSA-N 0.000 description 3
- PFTAWBLQPZVEMU-UKRRQHHQSA-N (-)-epicatechin Chemical group C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-UKRRQHHQSA-N 0.000 description 3
- 229930013783 (-)-epicatechin Natural products 0.000 description 3
- 235000007355 (-)-epicatechin Nutrition 0.000 description 3
- 239000003472 antidiabetic agent Substances 0.000 description 3
- 238000004587 chromatography analysis Methods 0.000 description 3
- 235000011949 flavones Nutrition 0.000 description 3
- IQTGAKWQIFFPQX-UHFFFAOYSA-N marsupsin Chemical compound O=C1C=2C(OC)=CC(O)=CC=2OC1(O)CC1=CC=C(O)C=C1 IQTGAKWQIFFPQX-UHFFFAOYSA-N 0.000 description 3
- OBWHQJYOOCRPST-UHFFFAOYSA-N 3,4',7-trihydroxyflavone Chemical compound C1=CC(O)=CC=C1C1=C(O)C(=O)C2=CC=C(O)C=C2O1 OBWHQJYOOCRPST-UHFFFAOYSA-N 0.000 description 2
- ACZGCWSMSTYWDQ-UHFFFAOYSA-N 3h-1-benzofuran-2-one Chemical class C1=CC=C2OC(=O)CC2=C1 ACZGCWSMSTYWDQ-UHFFFAOYSA-N 0.000 description 2
- LCAWNFIFMLXZPQ-UHFFFAOYSA-N 4',7-dihydroxyflavone Chemical compound C1=CC(O)=CC=C1C1=CC(=O)C2=CC=C(O)C=C2O1 LCAWNFIFMLXZPQ-UHFFFAOYSA-N 0.000 description 2
- HBFNWMJHTGIQRB-UHFFFAOYSA-N 4-[2-hydroxy-3-(4-hydroxyphenyl)propyl]benzene-1,3-diol Chemical compound C=1C=C(O)C=C(O)C=1CC(O)CC1=CC=C(O)C=C1 HBFNWMJHTGIQRB-UHFFFAOYSA-N 0.000 description 2
- CJJQLHLEWQGHMJ-UHFFFAOYSA-N 4-[2-hydroxy-3-(4-hydroxyphenyl)propyl]phenol Chemical compound C=1C=C(O)C=CC=1CC(O)CC1=CC=C(O)C=C1 CJJQLHLEWQGHMJ-UHFFFAOYSA-N 0.000 description 2
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 2
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000003178 anti-diabetic effect Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 239000012259 ether extract Substances 0.000 description 2
- XHEFDIBZLJXQHF-UHFFFAOYSA-N fisetin Chemical compound C=1C(O)=CC=C(C(C=2O)=O)C=1OC=2C1=CC=C(O)C(O)=C1 XHEFDIBZLJXQHF-UHFFFAOYSA-N 0.000 description 2
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N flavone Chemical compound O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 2
- 229930182478 glucoside Natural products 0.000 description 2
- 150000008131 glucosides Chemical group 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- JBQATDIMBVLPRB-UHFFFAOYSA-N isoliquiritigenin Natural products OC1=CC(O)=CC=C1C1OC2=CC(O)=CC=C2C(=O)C1 JBQATDIMBVLPRB-UHFFFAOYSA-N 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 230000008018 melting Effects 0.000 description 2
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 2
- JCPGMXJLFWGRMZ-UHFFFAOYSA-N 1-(2-hydroxyphenyl)-3-phenylpropan-1-one Chemical compound OC1=CC=CC=C1C(=O)CCC1=CC=CC=C1 JCPGMXJLFWGRMZ-UHFFFAOYSA-N 0.000 description 1
- KPWDGTGXUYRARH-UHFFFAOYSA-N 2,2,2-trichloroethanol Chemical compound OCC(Cl)(Cl)Cl KPWDGTGXUYRARH-UHFFFAOYSA-N 0.000 description 1
- XILIYVSXLSWUAI-UHFFFAOYSA-N 2-(diethylamino)ethyl n'-phenylcarbamimidothioate;dihydrobromide Chemical compound Br.Br.CCN(CC)CCSC(N)=NC1=CC=CC=C1 XILIYVSXLSWUAI-UHFFFAOYSA-N 0.000 description 1
- QLHSZVGLSUHUOI-UHFFFAOYSA-N 3,5,6-trihydroxy-2-phenylchromen-4-one Chemical compound OC=1C(=O)C2=C(O)C(O)=CC=C2OC=1C1=CC=CC=C1 QLHSZVGLSUHUOI-UHFFFAOYSA-N 0.000 description 1
- FBUPXHQDNXCVLC-UHFFFAOYSA-N 4',5-Di-Me ether,7-O-alpha-L-rhamnopyranoside-4',5,7-Trihydroxy-8-methylisoflavone Natural products COc1ccc(cc1)C2=COc3c(C)c(OC4OC(C)C(O)C(O)C4O)cc(OC)c3C2=O FBUPXHQDNXCVLC-UHFFFAOYSA-N 0.000 description 1
- QHCYSTGSLGXKMF-UHFFFAOYSA-N 4,6,4'-trihydroxy-7-methylaurone 4-O-rhamnopyranoside Natural products OC1C(O)C(O)C(C)OC1OC1=CC(O)=C(C)C(O2)=C1C(=O)C2=CC1=CC=C(O)C=C1 QHCYSTGSLGXKMF-UHFFFAOYSA-N 0.000 description 1
- ZWKNCUQZWIAEDC-UHFFFAOYSA-N 4,6,4'-trihydroxyaurone 6-O-rhamnopyranoside Natural products OC1C(O)C(O)C(C)OC1OC(C=C1O)=CC(O2)=C1C(=O)C2=CC1=CC=C(O)C=C1 ZWKNCUQZWIAEDC-UHFFFAOYSA-N 0.000 description 1
- 210000002237 B-cell of pancreatic islet Anatomy 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- IQTGAKWQIFFPQX-INIZCTEOSA-N Carpusin Natural products O(C)c1c2C(=O)[C@](O)(Cc3ccc(O)cc3)Oc2cc(O)c1 IQTGAKWQIFFPQX-INIZCTEOSA-N 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- YJHVMPKSUPGGPZ-UHFFFAOYSA-N Dihydro-beta-eudesmol Natural products C1CC(C(C)(C)O)CC2C(C)CCCC21C YJHVMPKSUPGGPZ-UHFFFAOYSA-N 0.000 description 1
- 241000220485 Fabaceae Species 0.000 description 1
- FNUPUYFWZXZMIE-UHFFFAOYSA-N Fustin Natural products O1C2=CC(O)=CC=C2C(=O)C(O)C1C1=CC=C(O)C(O)=C1 FNUPUYFWZXZMIE-UHFFFAOYSA-N 0.000 description 1
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 1
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 1
- FURUXTVZLHCCNA-UHFFFAOYSA-N Liquiritigenin Natural products C1=CC(O)=CC=C1C1OC2=CC(O)=CC=C2C(=O)C1 FURUXTVZLHCCNA-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 241000700157 Rattus norvegicus Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000012675 alcoholic extract Substances 0.000 description 1
- IPZIYGAXCZTOMH-UHFFFAOYSA-N alpha-eudesmol Natural products CC1=CCCC2CCC(CC12)C(C)(C)O IPZIYGAXCZTOMH-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 229940127003 anti-diabetic drug Drugs 0.000 description 1
- 229940125708 antidiabetic agent Drugs 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- XFSVWZZZIUIYHP-UHFFFAOYSA-N beta-Eudesmol Natural products CC(C)(O)C1CCC2CCCC(=C)C2C1 XFSVWZZZIUIYHP-UHFFFAOYSA-N 0.000 description 1
- BOPIMTNSYWYZOC-VNHYZAJKSA-N beta-eudesmol Chemical compound C1CCC(=C)[C@@H]2C[C@H](C(C)(O)C)CC[C@]21C BOPIMTNSYWYZOC-VNHYZAJKSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- PXLWOFBAEVGBOA-UHFFFAOYSA-N dihydrochalcone Natural products OC1C(O)C(O)C(CO)OC1C1=C(O)C=CC(C(=O)CC(O)C=2C=CC(O)=CC=2)=C1O PXLWOFBAEVGBOA-UHFFFAOYSA-N 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 239000012156 elution solvent Substances 0.000 description 1
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 1
- 239000000469 ethanolic extract Substances 0.000 description 1
- UREBWPXBXRYXRJ-UHFFFAOYSA-N ethyl acetate;methanol Chemical compound OC.CCOC(C)=O UREBWPXBXRYXRJ-UHFFFAOYSA-N 0.000 description 1
- NBEMQPLNBYYUAZ-UHFFFAOYSA-N ethyl acetate;propan-2-one Chemical compound CC(C)=O.CCOC(C)=O NBEMQPLNBYYUAZ-UHFFFAOYSA-N 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002213 flavones Chemical group 0.000 description 1
- WWULHQLTPGKDAM-UHFFFAOYSA-N gamma-eudesmol Natural products CC(C)C1CC(O)C2(C)CCCC(=C2C1)C WWULHQLTPGKDAM-UHFFFAOYSA-N 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 125000003147 glycosyl group Chemical group 0.000 description 1
- 125000000623 heterocyclic group Chemical group 0.000 description 1
- 208000021760 high fever Diseases 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 229940126904 hypoglycaemic agent Drugs 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- 229930013032 isoflavonoid Natural products 0.000 description 1
- 150000003817 isoflavonoid derivatives Chemical class 0.000 description 1
- 235000012891 isoflavonoids Nutrition 0.000 description 1
- DXDRHHKMWQZJHT-FPYGCLRLSA-N isoliquiritigenin Chemical compound C1=CC(O)=CC=C1\C=C\C(=O)C1=CC=C(O)C=C1O DXDRHHKMWQZJHT-FPYGCLRLSA-N 0.000 description 1
- 235000008718 isoliquiritigenin Nutrition 0.000 description 1
- FURUXTVZLHCCNA-AWEZNQCLSA-N liquiritigenin Chemical compound C1=CC(O)=CC=C1[C@H]1OC2=CC(O)=CC=C2C(=O)C1 FURUXTVZLHCCNA-AWEZNQCLSA-N 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- DHRLEVQXOMLTIM-UHFFFAOYSA-N phosphoric acid;trioxomolybdenum Chemical compound O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.OP(O)(O)=O DHRLEVQXOMLTIM-UHFFFAOYSA-N 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- VLEUZFDZJKSGMX-ONEGZZNKSA-N pterostilbene Chemical compound COC1=CC(OC)=CC(\C=C\C=2C=CC(O)=CC=2)=C1 VLEUZFDZJKSGMX-ONEGZZNKSA-N 0.000 description 1
- VLEUZFDZJKSGMX-UHFFFAOYSA-N pterostilbene Natural products COC1=CC(OC)=CC(C=CC=2C=CC(O)=CC=2)=C1 VLEUZFDZJKSGMX-UHFFFAOYSA-N 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 206010040872 skin infection Diseases 0.000 description 1
- GSZUGBAEBARHAW-UHFFFAOYSA-N sophoraflavone B Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C(C=2OC3=CC(O)=CC=C3C(=O)C=2)C=C1 GSZUGBAEBARHAW-UHFFFAOYSA-N 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 208000004371 toothache Diseases 0.000 description 1
- 208000019206 urinary tract infection Diseases 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
- Saccharide Compounds (AREA)
Description
【0001】
【産業上の利用分野】
本発明は新規な化合物、8‐(c‐β‐D‐グルコピラノシル)‐7,3’,4’‐トリヒドロキシフラヴォンに関する。また、本発明はキノノキ(Pterocarpus marsupium)から新規化合物8‐(c‐β‐D‐グルコピラノシル)‐7,3’,4’‐トリヒドロキシフラヴォンを分離する方法に関する。
【0002】
【従来の技術】
インド・キノノキ(Kino Tree)またはビジャサール(Bijasar)として知られるPterocarpus marsupium Roxb (マメ科)はインドの中央部の丘陵地域やインド半島でよく知られている(資料1)。この木の葉や花及び樹脂の抽出物は、下痢、歯痛、高熱、尿路感染症や皮膚感染症の治療に使用されてきた(資料2)。樹皮の抽出物は長い間糖尿病の治療に有効であるとみなされてきた(資料3)。チャクラヴァルティの報告によれば低血糖活性要素は(‐)‐エピカテキンであり、その効果は膵臓のベータ細胞の再生によるとされている(資料4)。しかしながら、この主張はコルブ等(資料5)及びシーハン等(資料6)により疑問とされてきた。現在では、人間の臨床研究に見込みのある坑糖尿病剤として(‐)‐エピカテキンを考慮する以前に、更なる研究が必要であると考えられている。
【0003】
インド・システム・オブ・メディシンの医者達は、キノノキの樹皮よりもむしろ心材の方が糖尿病患者の治療に有用であるという見解を持っている。心材のみが明瞭に赤色で、水に浸すと水を青緑色の蛍光を有する赤色に着色する。これが坑糖尿病薬として使用するのに適していると言う主張でもある。キノノキ心材からの水またはアルコール抽出物の低血糖効果は実験的に証明されており(資料7、8)、臨床研究においても証明されている(資料9,10)。キノノキの心材は多くのフェノール類を含んでいる。キノノキ心材の化学的研究は1946年に遡るが、この薬品に関する初期の研究は断片的なものであった(資料11)。
【0004】
以前に報告されているこの植物に関する研究には以下の化学的成分が開示されている。
【0005】
1.キノノキ心材のエーテル抽出物からは、マルスポール(Marusupol)と名づけられているイソフラヴォノイド・グリコール4,4'‐ジヒドロキシ‐α‐メチルヒドロベンゾイン(資料12)、ベンゾフラノンの誘導体でカルプシン(Carpusin)と命名されている2,4',6‐トリヒドロキシ‐4‐メトキシベンゾ(b)フラン3(2H)‐1(資料13)、2‐プロパノール誘導体でプロプテロールと命名されている1,3‐ビス(4‐ヒドロキシフェニル)プロパン‐2‐オール(資料14)、プロプテロールBと命名されている1‐(2,4‐ジヒドロキシフェニル)‐3‐(4‐ヒドロキシフェニル)プロパン‐2‐オール(資料15)、6‐ヒドロキシ‐7‐O‐メチル‐3‐(3‐ヒドロキシ‐4‐O‐メチル ベンジル)クロマン‐4‐1(資料16)が得られている。
【0006】
2.心材のアルコール抽出物のエチルアセテート溶解成分からは、ペトロスピンβ、2',4,4',‐テトラヒドロキシ‐3'(c‐β‐D‐グルコピラノシド)ジヒドロ カルコン(資料17)、マルスピノール(Marsupinol)(資料18)、5,4'‐ジメトキシ‐8‐メチルイソフラヴォン‐7‐O‐α‐L‐ラムノピラノシド、retusi n‐O‐β‐D‐グルコピラノシド及びイリソリディン‐7‐O‐α‐L‐ラムノピ ラノシド(資料19)、5,7'‐ジヒドロキシ‐6‐メトキシ‐7‐O‐α‐L‐ ラムノピラノシド(資料20)を得ている。
【0007】
3.脱脂した心材のエチルアセテート抽出物からは、ベンゾフラノン誘導体、2,6‐ジヒドロキシ‐2‐(p‐ヒドロキシルベンジル)‐4‐メトキシ‐3(2H)‐ベンゾフラノン(マルスピンと命名)(資料21)、プテロスチルビン、(2S)‐ヒドロキシフラヴォン、イソリキリチゲニン、リキリチゲニン、7,4'‐ジヒドロキシフラヴォン、5‐デオキシカンフェロール 及び3,7,4'‐トリヒドロキシフラヴォン(資料22)、2つのグリコシド、8‐C‐β‐D‐グルコピラノシル‐3,7,4'‐トリヒドロキシと、3,7,3',4'‐テトラヒドロキシフラヴォンと、3'‐C‐β‐D‐グルコピラノシル‐α‐ヒドロキシ・ジヒドロカルコン(資料23)を得ている。
【0008】
4.キノノキの根のガソリン抽出物からは、セリン‐4(15)‐ene‐1β、11‐ジオール、β‐オイデスモール、エリスロジオール‐3‐モノアセテート及びプテロスチルベン(資料24)を得ることができる。キノノキの花のエタノール抽出物からは、4,6,4'‐トリヒドロキシアウロン6‐O‐ラムノピラノシド及び4,6,4'‐トリヒドロキシ‐7‐メチルアウロン‐4‐O‐ラムノピラノシド(資料25)をえている。キノノキの樹皮のエタノール抽出物からは、(‐)‐エピカテキン(資料26)を得ている。
【0009】
しかしながら、これら先行技術はかかる化学成分に関連する生物学的活性について詳細を提供していない。また、これら先行技術はエーテル抽出物、エチルアセテート抽出物及びアルコール抽出物のエチルアセテート溶解成分の調製についてのみを開示し、キノノキ心材の水による抽出物の調製法及び当該抽出物から如何なる化学成分の分離の試みも開示していない。
【0010】
【発明が解決しようとする課題】
したがって、本発明の主たる目的は、キノノキの心材の水抽出物を調製することであり、その抽出物から化学成分を得ることである。本発明の他の目的はキノノキの心材の水抽出物を研究して糖尿病の治療に有用な生物学的活性のある成分を得ることにある。
【0011】
【課題を解決するための手段】
本発明の上記及びその他の目的は、キノノキの心材の水抽出物のn‐ブタノール溶解成分を調製することによって、更に、それより新規な生物学的活性成分を分離することにより達成される。したがって、本発明は新規な化合物、8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォン{8-(C-β-D-glucopyranosyl)-7,3',4'-trihydroxyflavone}を提供する。
【0012】
また、本発明は、8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンを分離するための下記のステップからなる方法を提供する。(a)植物、キノノキ(Pterocarpus marsupium)の心材を粉末にする、(b)プロトン性溶媒を用いて前記粉末化植物から抽出する、(c)抽出物を最小容量に濃縮し、異なる有機溶媒により分離して非極成分を除去し、(d)残留物から8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンを分離する。
【0013】
本発明の一実施形態においては、ステップ(b)で抽出物の調製に使用するプロトン性溶媒は水、メタノール、エタノール,プロパノール,ブタノール及びそれらの混合物からなる群から選択される。更に,本発明の実施形態においては、水の層に抽出するのに使用する極性溶媒はエチルアセテート、プロパノール、ブタノールから選択される。本発明の他の実施形態においては、ステップ(c)において非極性成分の除去に使用する有機溶媒はへキサン、石油エーテル及びクロロフォルムからなる群から選択される。本発明の他の実施形態においては、8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンの分離に使用するクロマトグラフィー法は中圧液体クロマトグラフィー(MPLC)、高速液体クロマトグラフィー(HPLC)、フラッシュクロマトグラフィーから選択される。
【0014】
本発明はまた薬物として許容できるキャリア中に薬物として有効な量の8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンを含有する薬物組成物に関する。本発明の一実施形態においては、前記薬物組成物中の8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンの量は,患者の体重1kg当たり0.5から10mgの範囲である。
【0015】
本発明はまた糖尿病の治療のための薬物組成物の調製に8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンを用いることに関する。
【0016】
【発明の実施の形態】
本発明は、8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンを分離する方法を提供し、その方法は下記ステップよりなる。
(a)キノノキ(Pterocarpus marsupium)の心材を粉末にする、
(b)粉末化した植物原料をプロトン性溶媒を用いて抽出する、
(c)水性抽出物を最小容積まで濃縮し、無極性成分を除去するため極性を増加する有機溶媒で分離し、極性溶媒により水層に抽出する、溶媒を除去し残留物を得る、
(d)残留物から8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンを分離する。
【0017】
抽出物を調製するため使用する溶媒は、水、メタノール、エタノール、プロパノール、ブタノール、またはそれらの混合物である。無極性成分を除去するためにステップ(c)で使用する有機溶媒は,へキサン,石油エーテル及びクロロフォルムからなる群より選択される。水性層に抽出するために使用する極性溶媒は,エチルアセテート、プロパノールまたはブタノールから選択される。8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンの分離に使用するクロマトグラフィは中圧液体クロマトグラフィ、フラッシュクロマトグラフィ等であってよい。中圧液体クロマトグラフィにおいて必要な溶離溶媒はカラムを通じてポンプにより圧送され、フラッシュクロマトグラフィにおいては溶媒は空気圧により圧送される。前記化合物は分子式C21H20O10(FAB質量分析法により、m/z433[M+1])である。この結果は13C核磁気共鳴及びDEPTスペクトルによって支持された。
【0018】
化合物8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンはキノノキの心材からの水による浸出液のn‐ブタノール溶解成分から分離され、人間及び動物において抗糖尿病活性を示す。当該技術分野でなされてきた研究は、エチルアセテート抽出物やアルコール抽出物のエチルアセテート溶解成分についてであり、前記化合物に付いては先行技術に開示がない。
【0019】
キノノキから活性要素を分離する方法は、粉末化した心材の水による抽出物から分子中に炭素数1〜6を含む種々の有機溶媒を用いて分離することからなる。8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンは、極性成分から中圧液体クロマトグラフィ(MPLC)、高速液体クロマトグラフィ(HPLC)またはシリカゲル(230〜400メッシュ)を用いたフラッシュクロマトグラフィ等の近代的なクロマトグラフィ技術を適用して分離され、低血糖活性を示す。
【0020】
化合物8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンは、18時間断食させたウイスター鼠(Wisterrat)で低糖尿活性が評価されている。10mg/kg(体重)の処方量で、全ての鼠に低血糖効果が記録されている。投与前の平均92mg/血液100mlから平均68mg/血液100mlまで、平均降下量は24mg/血液100mlを記録している。これに比して、研究において対照テストに採用された低血糖剤の平均降下量は23mg/血液100mlであった。
【0021】
前記化合物は,リンモリブデン酸テスト(青色)や塩化第二鉄反応(緑色)によりフェノールであることが証明されている。Shinodaテストの反応からフラヴォンであることも認められている。ヒドロキシル基、カルボニル基及びフェニル核の存在が、3228、1615,1554、1448、1422cm‐1の赤外線吸収により示されている。この化合物の紫外線吸収スペクトルは、λmax MeOH219、238、260、320、358nmに最大吸収を示し、NaOAcの存在で219、238、267、320,367nmに深色移動する。この観察はC−7に位置するフリーのヒドロキシル基が存在することを示唆する。
【0022】
1H核磁気共鳴スペクトル(200MHz、ジメチルスルフォキシド‐d6)は、グルコシルとB環による立体的高密度の故と想定されるが、芳香族領域において信号が広がりを示す。スペクトル検査によりフラボンのC−3位置のプロトンの特徴であるδ6.98(1H)の一重信号が現れる。δ8.28(1H、d、J=2.1Hz)の二重プロトン信号は、C=Oの近傍効果により低磁場シフトしているが、δ6.95(1H、d、J=8.3Hz)の二重信号とオルソ結合している。このオルソ結合はC−5及びC−6のプロトンによるものであり、これら2つのプロトンのみがA環に帰属し、C−8はグリコシル基に含まれていることを示している。δ7.84(1H、br、J=2.1Hz)、7.97(1H,brdd、J=2.1,8.7Hz)、及び6.99(1H,d,J=8.7Hz)のプロトンの信号はB環のプロトンによる。更に、1H及び13C核磁気共鳴スペクトルはグルコース構成成分によるものである。C−C結合はδ5.16のアノマープロトンとグルコシド置換C−の特性領域におけるδ79.3の二重炭素との1H及び13C異核分子相関作用により実証された。更に、グルコピラノシドのアノマープロトンに帰結する信号の結合定数(J=9.5Hz)はグルコシドリンクがβ配置であることを示した。かくして,上記解析により構造が8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンである結論に達した。
【0023】
本発明の詳細について以下に実施例を参照して記載するが、実施例は本発明における技術範囲の限定を構成するものではない。
【0024】
実施例1
キノノキの心材を粉末にしたもの(1kg)を80%のエタノール水(3×3l)を用いて48時間抽出した。抽出後濃縮したものをヘキサン,クロロフォルム、プロパノール、ブタノールで順番に分離した。極性抽出物はヘキサン、クロロフォルム、メタノール,エタノールを順次使用し、シリカゲル(100〜200メッシュ)による中圧液体クロマトグラフィを行った。活性化合物は中圧液体クロマトグラフィと、トリクロロメチル‐メタノール(19:1)を溶媒としシリカゲル(230〜400メッシュ)によるフラッシュクロマトグラフィとを繰り返し行い精製して、8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォン(収率0.046%)、融点202〜204℃、[α]D 19+25.6°(メタノール,c,0.5)を得た。
【0025】
実施例2
キノノキの心材を温水で4×4時間抽出した。抽出濃縮物をヘキサン、クロロフォルム、プロパノール及びブタノールで順次溶解分離した。得られた極性抽出物からヘキサン、クロロフォルム、エチルアセテート,メタノールを溶媒システムとしてシリカゲル(100〜200メッシュ)によるフラッシュクロマトグラフィにより8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンに富む成分を得た。該成分をEtOAc‐MeOH(19.5:0.5)を溶媒としシリカゲル(230〜400メッシュ)によるクロマトグラフィを繰り返して、分子式8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォン(収率0.049%)、融点202〜204℃、[α]D 19+25.6°(メタノール,c,0.5)を得た。
【0026】
実施例3
キノノキ心材を水の容積が1/4になるまで水と共に煮沸した(16回)。濾過し濃縮してヘキサン、クロロフォルム、エチルアセテート,プロパノール及びn‐ブタノールを用いて順次分離した。得られた極性抽出物からヘキサン、クロロフォルム、エチルアセテート及びメタノールを溶媒システムに用いてシリカゲル(60〜120メッシュ)のカラムクロマトグラフィにより8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンに富む成分を得た。当該8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンに富む成分をエチルアセテート‐アセトン(8:2)の混合溶媒を用いてシリカゲル(100〜200メッシュ)のカラムクロマトグラフィを繰り返し8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォン(収率0.051%)、融点202〜204℃、[α]D 19+25.6°(メタノール,c,0.5)を得た。
【0027】
【発明の効果】
1.得られた化合物8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンは抗糖尿病活性を有する新しい分子構造の化合物である。
2.8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンの分離方法は比較的簡単である。
【0028】
【参考資料】
1.Jain, S.K.Medicinal Plants, National Bank trust, New Delhi, 1968, p. 116
2. Chopra,R.N.,Chopra, I.C., Handa, K.L. and Kapur, L.D. Indigenous Drugs of India, 2nd Ed., Dhar, U.N. and Sons Private Limited, Calcutta, 1958, p. 522
3.Kirtikar, K.R. and Basu, B.D.Indian Medicinal Plants, 2nd Ed. Edited by Blatter, E., Cailes, J.F. and Mhaskar, K.S. Sinbh and Singh, Delhi. India, 1975,p. 2135
4.Chakravarthy, B.K., Gupta, S. and Gode, K.D., Lancet, 1982, 272 (cited references)
5.Kolb, H., Kiesel, U., Grenlich, B. and Bosch, J.V.D. Lancrt, 1982, 1303
6.Sheehan, E.W., Zemaitis, M.A., Slatkin, D.J. and Schiff, Jr. P.L. Journal of Natural Products, 1983, 46, 232
7.Shah, D.S. Indian Journal of Medical Research, 1967,55, 166 (cited references)
8.Gupta, S.S. Indian Journal of Medical Research, 1963,51, 716
9.Sepha, G.C. and Bose, S.N. J. Ind. Med. Assoc. 1956, 27, 383
10.Kedar, P. and Chkrabarti, C.H., Maharastra Med. J., 1981, 28, 165
11.Bhargava, P.N., Proc. Ind. Acad. Sci., 1946, 24A, 496
12.Rao, A.V.S., Mathew, J., Phytochemistry, 1982, 21, 1837
13.Mathew, J. and Rao, A.V.S. Phytochemistry, 1983, 22, 794
14.Rao, A.V.S., Mathew, J. and Shankaran, A.V.B. Phytochemistry, 1984, 23, 897
15.Mathew, J., Rao, A.V.S. and Rambhav, S.Current Science, 1984, 53, 576
16.Jain, S.C., Sharma, S.K., Kumar, R., Rajwansh, V.K. and Babu, V.R. Phytochemistry, 1997, 44, 765
17.Adinarayana, D., Syamsundar, K.V., Seligmann, O. and Wagner, H.
Z. Naturforsch. 1982, 37C, 145
18.Trivedi, J.J. Indian J. Phys. Pharmacol. 1997, 15, 51
19.Mitra, J. and Joshi, T. Phytochemistry, 1982, 21, 2429
20.Mitra, J. and Joshi, T. Phytochemistry, 1983, 22, 2326
21.Maurya, R., Ray, A.B., Duah, F.K., Slatkin, D.J. and Schiff, P.L.Jr. Heterocycles 1982, 19, 2103
22.Maurya, R., Ray, A.B., Duah, F.K., Slatkin, D.J. and Schiff, P.L.Jr.J. Nat. Prod. 1984, 47, 179
23. Bezuidenhoudt, B.C.B., Brandt, E.V. and Ferreira, E.V.
Phytochemistry, 1987, 26, 531
24.Adinarayana, D. and Syamasundar, K.V. Phytochemistry, 1982, 22, 1082
25. Mohan, P. and Joshi, T. Phytochemistry, 1989, 28, 1287
26. Chakravarthy, B.K. and Gode, K.D. Planta Medica 1985, 56[0001]
[Industrial application fields]
The present invention relates to a novel compound, 8- (c-β-D-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone. The present invention also relates to a method for separating the novel compound 8- (c-β-D-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone from Pterocarpus marsupium .
[0002]
[Prior art]
Pterocarpus marsupium Roxb (Leguminosae), known as the Indian Kinoki or Bijasar, is well known in the central hills of India and the Indian Peninsula (Appendix 1). The leaves, flowers and resin extracts have been used to treat diarrhea, toothache, high fever, urinary tract infections and skin infections (Document 2). Bark extracts have long been considered effective in the treatment of diabetes (Appendix 3). According to the report of Chakravarty, the hypoglycemic activity factor is (−)-epicatechin, and its effect is due to regeneration of pancreatic beta cells (Reference 4). However, this allegation has been questioned by Kolb et al. (Document 5) and Sheehan et al. (Document 6). At present, further studies are considered necessary before considering (-)-epicatechin as a potential anti-diabetic agent for human clinical research.
[0003]
Indian System of Medicine doctors have the view that heartwood is more useful in the treatment of diabetics than cypress bark. Only the heartwood is clearly red, and when immersed in water, the water is colored red with blue-green fluorescence. It is also a claim that this is suitable for use as an antidiabetic drug. The hypoglycemic effect of water or alcoholic extracts from cypress heartwood has been experimentally proven (Documents 7 and 8) and has also been demonstrated in clinical studies (Documents 9 and 10). Mushroom heartwood contains many phenols. Although chemical research on quinoki heartwood dates back to 1946, early research on this drug was fragmented (Appendix 11).
[0004]
Previously reported studies on this plant disclosed the following chemical components:
[0005]
1. From the ether extract of cypress heartwood, isoflavonoid glycol 4,4'-dihydroxy-α-methylhydrobenzoin (Mar. 12) named Marusupol, a derivative of benzofuranone and Carpusin 2,4 ′, 6-Trihydroxy-4-methoxybenzo (b) furan 3 (2H) -1 (Appendix 13), named 1,3-bis (Propterol, a 2-propanol derivative 4-Hydroxyphenyl) propan-2-ol (Document 14), 1- (2,4-dihydroxyphenyl) -3- (4-hydroxyphenyl) propan-2-ol, named Propterol B (Document 15) 6-Hydroxy-7-O-methyl-3- (3-hydroxy-4-O-methyl benzyl) chroman-4-1 (Document 1) ) Has been obtained.
[0006]
2. From the ethyl acetate-dissolved component of the heartwood alcohol extract, petrospin β, 2 ′, 4,4 ′,-tetrahydroxy-3 ′ (c-β-D-glucopyranoside) dihydrochalcone (Document 17), Malspinol ( Marsupinol (Document 18), 5,4'-dimethoxy-8-methylisoflavone-7-O-α-L-rhamnopyranoside, retusin-O-β-D-glucopyranoside and irisolidin-7-O-α-L -Rhamnopyranoside (Reference 19), 5,7'-dihydroxy-6-methoxy-7-O-α-L-rhamnopyranoside (Reference 20).
[0007]
3. From the ethyl acetate extract of defatted heartwood, a benzofuranone derivative, 2,6-dihydroxy-2- (p-hydroxylbenzyl) -4-methoxy-3 (2H) -benzofuranone (named Marspin) (Document 21), Ptero Stilbin, (2S) -hydroxyflavone, isoliquiritigenin, liquiritigenin, 7,4'-dihydroxyflavone, 5-deoxycamphorol and 3,7,4'-trihydroxyflavone (Document 22), 2 Two glycosides, 8-C-β-D-glucopyranosyl-3,7,4'-trihydroxy, 3,7,3 ', 4'-tetrahydroxyflavone and 3'-C-β-D-glucopyranosyl -Α-Hydroxy dihydrochalcone (Document 23) is obtained.
[0008]
4). Serine-4 (15) -ene-1β, 11-diol, β-eudesmol, erythrodiol-3-monoacetate and pterostilbene (Document 24) can be obtained from gasoline extract of cypress root. 4,6,4'-Trihydroxyaurone 6-O-rhamnopyranoside and 4,6,4'-trihydroxy-7-methylaurone-4-O-rhamnopyranoside (Reference 25) Have (-)-Epicatechin (Document 26) is obtained from the ethanol extract of the bark of the mushroom.
[0009]
However, these prior arts do not provide details on the biological activity associated with such chemical components. Also, these prior arts only disclose the preparation of ether extract, ethyl acetate extract and ethyl acetate soluble components of alcohol extract, and the method of preparing the extract of Kinoki heartwood with water and any chemical components from the extract. No attempt to separate is disclosed.
[0010]
[Problems to be solved by the invention]
Therefore, the main object of the present invention is to prepare an aqueous extract of cypress heartwood and to obtain chemical components from the extract. Another object of the present invention is to study an aqueous extract of cypress heartwood to obtain biologically active ingredients useful in the treatment of diabetes.
[0011]
[Means for Solving the Problems]
The above and other objects of the present invention are achieved by preparing an n-butanol soluble component of an aqueous extract of cypress heartwood and further isolating a new biologically active component therefrom. Accordingly, the present invention provides a novel compound, 8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone {8- (C-β-D-glucopyranosyl) -7,3 ′, 4 '-trihydroxyflavone} is provided.
[0012]
The present invention also provides a method comprising the following steps for separating 8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone. (A) plant, Pterocarpus marsupium heartwood is powdered, (b) extracted from the powdered plant using a protic solvent, (c) the extract is concentrated to a minimum volume, and with different organic solvents Separate to remove non-polar components and (d) separate 8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone from the residue.
[0013]
In one embodiment of the invention, the protic solvent used in the preparation of the extract in step (b) is selected from the group consisting of water, methanol, ethanol, propanol, butanol and mixtures thereof. Furthermore, in an embodiment of the invention, the polar solvent used to extract into the water layer is selected from ethyl acetate, propanol, butanol. In another embodiment of the present invention, the organic solvent used to remove non-polar components in step (c) is selected from the group consisting of hexane, petroleum ether and chloroform. In other embodiments of the present invention, the chromatographic method used to separate 8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone is medium pressure liquid chromatography (MPLC), Selected from high performance liquid chromatography (HPLC), flash chromatography.
[0014]
The invention also effective amounts of drug in a carrier acceptable as a drug 8- (C-beta-glucopyranosyl) -7,3 ', it relates to a drug composition containing 4'-trihydroxy hula Von. In one embodiment of the invention, the amount of 8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone in the drug composition is from 0.5 to 1 kg of patient body weight. The range is 10 mg.
[0015]
The present invention is also 8 for the preparation of pharmaceutical composition for the treatment of diabetes (C-beta-glucopyranosyl) -7,3 ', relates to the use of 4'-trihydroxy hula Von.
[0016]
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides a method for separating 8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone, which method comprises the following steps.
(A) Powdering the heartwood of Pterocarpus marsupium,
(B) extracting the powdered plant material using a protic solvent;
(C) Concentrate the aqueous extract to a minimum volume, separate with an organic solvent of increasing polarity to remove non-polar components, extract into an aqueous layer with a polar solvent, remove the solvent to obtain a residue,
(D) Separating 8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone from the residue.
[0017]
The solvent used to prepare the extract is water, methanol, ethanol, propanol, butanol, or mixtures thereof. The organic solvent used in step (c) to remove non-polar components is selected from the group consisting of hexane, petroleum ether and chloroform. The polar solvent used for extraction into the aqueous layer is selected from ethyl acetate, propanol or butanol. The chromatography used for the separation of 8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone may be medium pressure liquid chromatography, flash chromatography or the like. The elution solvent required in medium pressure liquid chromatography is pumped through a column by a pump, and in flash chromatography, the solvent is pumped by air pressure. The compound has the molecular formula C 21 H 20 O 10 (m / z 433 [M + 1] by FAB mass spectrometry). This result was supported by 13 C nuclear magnetic resonance and DEPT spectra.
[0018]
Compound 8- (C-β-glucopyranosyl) -7,3 ', 4'-trihydroxyflavone is isolated from n-butanol soluble component of water leachate from cypress heartwood and has anti- diabetic activity in humans and animals Show. The research that has been conducted in this technical field is about ethyl acetate extract components of ethyl acetate extract and alcohol extract, and there is no disclosure in the prior art about the compounds.
[0019]
The method for separating the active element from the mushroom consists of separating the powdered heartwood water extract using various organic solvents containing 1 to 6 carbon atoms in the molecule. 8- (C-β-Glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone can be converted from polar components to medium pressure liquid chromatography (MPLC), high performance liquid chromatography (HPLC) or silica gel (230-400 mesh). It is isolated by applying modern chromatography techniques such as flash chromatography used and exhibits hypoglycemic activity.
[0020]
The compound 8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone has been evaluated for low diabetic activity in Wistarrat fasted for 18 hours. A hypoglycemic effect has been recorded in all wrinkles at a prescription of 10 mg / kg (body weight). From an average of 92 mg / 100 ml of blood before administration to an average of 68 mg / 100 ml of blood, the average drop is recorded as 24 mg / 100 ml of blood. In comparison, the average drop in hypoglycemic agent employed in the control test in the study was 23 mg / 100 ml blood.
[0021]
The compound has been proven to be phenol by the phosphomolybdic acid test (blue) and ferric chloride reaction (green). It is also recognized as a flavon from the reaction of the Shinoda test. The presence of hydroxyl, carbonyl and phenyl nuclei is indicated by infrared absorption at 3228, 1615, 1554, 1448 and 1422 cm- 1 . The ultraviolet absorption spectrum of this compound shows maximum absorption at λ max MeOH 219, 238, 260, 320, 358 nm, and moves deeply to 219, 238, 267, 320, 367 nm in the presence of NaOAc. This observation suggests that there is a free hydroxyl group located at C-7.
[0022]
The 1 H nuclear magnetic resonance spectrum (200 MHz, dimethyl sulfoxide-d 6 ) is assumed to be due to steric high density due to glucosyl and the B ring, but the signal shows broadening in the aromatic region. Spectral examination reveals a single signal of δ6.98 (1H), which is characteristic of protons at the C-3 position of flavones. The double proton signal of δ8.28 (1H, d, J = 2.1 Hz) is shifted by a low magnetic field due to the proximity effect of C = O, but δ6.95 (1H, d, J = 8.3 Hz). Ortho-coupled to the dual signal. This ortho bond is due to the C-5 and C-6 protons, indicating that only these two protons belong to the A ring and C-8 is contained in the glycosyl group. δ 7.84 (1H, br, J = 2.1 Hz), 7.97 (1H, brdd, J = 2.1, 8.7 Hz), and 6.99 (1H, d, J = 8.7 Hz) The proton signal is derived from the B ring proton. Furthermore, 1 H and 13 C nuclear magnetic resonance spectra are due to glucose constituents. C—C bonds were demonstrated by 1 H and 13 C heteronuclear correlations between the anomeric proton of δ 5.16 and the double carbon of δ 79.3 in the characteristic region of glucoside substituted C-. Furthermore, the binding constant of the signal resulting from the anomeric proton of glucopyranoside (J = 9.5 Hz) indicated that the glucoside link is in the β configuration. Thus, the above analysis led to the conclusion that the structure is 8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone.
[0023]
Details of the present invention will be described below with reference to examples, but the examples do not constitute limitations on the technical scope of the present invention.
[0024]
Example 1
A powder (1 kg) of mushroom heartwood was extracted with 80% ethanol water (3 × 3 l) for 48 hours. The concentrated product after extraction was separated with hexane, chloroform, propanol and butanol in order. For the polar extract, hexane, chloroform, methanol, and ethanol were sequentially used, and medium pressure liquid chromatography using silica gel (100 to 200 mesh) was performed. The active compound was purified by repeated medium pressure liquid chromatography and flash chromatography on silica gel (230-400 mesh) using trichloromethyl-methanol (19: 1) as a solvent, and 8- (C-β-glucopyranosyl) -7 , 3 ′, 4′-trihydroxyflavone (yield 0.046%), melting point 202-204 ° C., [α] D 19 + 25.6 ° (methanol, c, 0.5).
[0025]
Example 2
Mushroom heartwood was extracted with warm water for 4 × 4 hours. The extract concentrate was dissolved and separated sequentially with hexane, chloroform, propanol and butanol. 8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxy was obtained by flash chromatography on silica gel (100 to 200 mesh) using hexane, chloroform, ethyl acetate and methanol as solvent systems from the obtained polar extract. Obtained flavon-rich ingredients. The component was repeatedly chromatographed on silica gel (230-400 mesh) using EtOAc-MeOH (19.5: 0.5) as a solvent to obtain the molecular formula 8- (C-β-glucopyranosyl) -7,3 ′, 4′- Trihydroxyflavone (yield 0.049%), melting point 202-204 ° C., [α] D 19 + 25.6 ° (methanol, c, 0.5) was obtained.
[0026]
Example 3
Butterberries were boiled with water until the water volume was reduced to 1/4 (16 times). Filtration, concentration and sequential separation using hexane, chloroform, ethyl acetate, propanol and n-butanol. The obtained polar extract was subjected to column chromatography on silica gel (60 to 120 mesh) using hexane, chloroform, ethyl acetate and methanol as a solvent system to obtain 8- (C-β-glucopyranosyl) -7,3 ′, 4′- A component rich in trihydroxyflavone was obtained. The 8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone-rich component was mixed with silica gel (100-200 mesh) using a mixed solvent of ethyl acetate-acetone (8: 2). Repeated column chromatography 8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone (0.051% yield), mp 202-204 ° C., [α] D 19 + 25.6 ° (Methanol, c, 0.5) was obtained.
[0027]
【The invention's effect】
1. The resulting compound 8- (C-beta-glucopyranosyl) -7,3 ', 4'-trihydroxy hula Von is a compound of new molecular structures having anti-diabetic activity.
The method for separating 2.8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone is relatively simple.
[0028]
【Reference document】
1.Jain, SKMedicinal Plants, National Bank trust, New Delhi, 1968, p. 116
2. Chopra, RN, Chopra, IC , Handa, KL and Kapur, LD Indigenous Drugs of India, 2 nd Ed., Dhar, UN and Sons Private Limited, Calcutta, 1958, p. 522
3.Kirtikar, KR and Basu, BDIndian Medicinal Plants, 2 nd Ed. Edited by Blatter, E., Cailes, JF and Mhaskar, KS Sinbh and Singh, Delhi. India, 1975, p. 2135
4.Chakravarthy, BK, Gupta, S. and Gode, KD, Lancet, 1982, 272 (cited references)
5.Kolb, H., Kiesel, U., Grenlich, B. and Bosch, JVD Lancrt, 1982, 1303
6.Sheehan, EW, Zemaitis, MA, Slatkin, DJ and Schiff, Jr. PL Journal of Natural Products, 1983, 46, 232
7.Shah, DS Indian Journal of Medical Research, 1967,55, 166 (cited references)
8.Gupta, SS Indian Journal of Medical Research, 1963,51, 716
9.Sepha, GC and Bose, SNJ Ind. Med.Assoc. 1956, 27, 383
10.Kedar, P. and Chkrabarti, CH, Maharastra Med. J., 1981, 28, 165
11.Bhargava, PN, Proc. Ind. Acad. Sci., 1946, 24A, 496
12.Rao, AVS, Mathew, J., Phytochemistry, 1982, 21, 1837
13.Mathew, J. and Rao, AVS Phytochemistry, 1983, 22, 794
14.Rao, AVS, Mathew, J. and Shankaran, AVB Phytochemistry, 1984, 23, 897
15.Mathew, J., Rao, AVS and Rambhav, S. Current Science, 1984, 53, 576
16.Jain, SC, Sharma, SK, Kumar, R., Rajwansh, VK and Babu, VR Phytochemistry, 1997, 44, 765
17.Adinarayana, D., Syamsundar, KV, Seligmann, O. and Wagner, H.
Z. Naturforsch. 1982, 37C, 145
18.Trivedi, JJ Indian J. Phys. Pharmacol. 1997, 15, 51
19.Mitra, J. and Joshi, T. Phytochemistry, 1982, 21, 2429
20.Mitra, J. and Joshi, T. Phytochemistry, 1983, 22, 2326
21.Maurya, R., Ray, AB, Duah, FK, Slatkin, DJ and Schiff, PLJr. Heterocycles 1982, 19, 2103
22.Maurya, R., Ray, AB, Duah, FK, Slatkin, DJ and Schiff, PLJr.J.Nat.Prod.1984, 47, 179
23. Bezuidenhoudt, BCB, Brandt, EV and Ferreira, EV
Phytochemistry, 1987, 26, 531
24.Adinarayana, D. and Syamasundar, KV Phytochemistry, 1982, 22, 1082
25. Mohan, P. and Joshi, T. Phytochemistry, 1989, 28, 1287
26. Chakravarthy, BK and Gode, KD Planta Medica 1985, 56
Claims (11)
(a)植物キノノキの心材を粉末化し、
(b)粉末化した植物原料の抽出物をプロトン性溶媒を用いて調製し、
(c)前記抽出物を最小容積まで濃縮し、無極性成分を除去し極性を増加する異なる有機溶媒を用いて分離し、極性溶媒により水性層に抽出し、溶媒を除去して残留物を得る、
(d)前記残留物から8‐(C‐β‐グルコピラノシル)‐7,3',4'‐トリヒドロキシフラヴォンを分離する、以上のステップからなる分離方法。8- (C-beta-glucopyranosyl) -7,3 ', a method of separating 4'-trihydroxy hula von ing ones quality,
(A) Powdering the heartwood of plant mushrooms,
(B) preparing a pulverized plant raw material extract using a protic solvent,
(C) Concentrate the extract to a minimum volume, remove non-polar components and separate using different organic solvents that increase polarity, extract to aqueous layer with polar solvent, remove solvent to obtain residue ,
(D) A separation method comprising the above steps, wherein 8- (C-β-glucopyranosyl) -7,3 ′, 4′-trihydroxyflavone is separated from the residue.
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/IN2002/000085 WO2003082887A1 (en) | 2002-03-28 | 2002-03-28 | 8-(c-beta-d-glucopyranosyl)-7,3',4'-trihydroxyflavone, process of isolation from pterocarpus marsupium and pharmaceutical composition for the treatment of diabetes |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2005519979A JP2005519979A (en) | 2005-07-07 |
JP3943082B2 true JP3943082B2 (en) | 2007-07-11 |
Family
ID=34044315
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2003580351A Expired - Fee Related JP3943082B2 (en) | 2002-03-28 | 2002-03-28 | Substance having molecular structure of 8- (c-β-D-glucopyranosyl) -7,3 ', 4'-trihydroxyflavone, separation method thereof and drug composition using the same |
Country Status (3)
Country | Link |
---|---|
JP (1) | JP3943082B2 (en) |
AT (1) | ATE286905T1 (en) |
AU (1) | AU2002249555A1 (en) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP6551924B2 (en) * | 2015-05-25 | 2019-07-31 | 学校法人近畿大学 | Periodontal disease remedy and method for producing the same |
CN112480196A (en) * | 2020-12-03 | 2021-03-12 | 中国农业科学院蜜蜂研究所 | Hypoglycemic component in camellia bee pollen and extraction method and application thereof |
-
2002
- 2002-03-28 AT AT02718508T patent/ATE286905T1/en not_active IP Right Cessation
- 2002-03-28 AU AU2002249555A patent/AU2002249555A1/en not_active Abandoned
- 2002-03-28 JP JP2003580351A patent/JP3943082B2/en not_active Expired - Fee Related
Also Published As
Publication number | Publication date |
---|---|
JP2005519979A (en) | 2005-07-07 |
AU2002249555A1 (en) | 2003-10-13 |
ATE286905T1 (en) | 2005-01-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US6562791B1 (en) | Glucopyranoside, process for isolation thereof, pharmaceutical composition containing same and use thereof | |
US6617313B1 (en) | Glucopyranoside and process of isolation thereof from pterocarpus marsupium pharmaceutical composition containing the same and use thereof | |
EP1381619B1 (en) | 8-(c-beta-d-glucopyranosyl)-7,3',4'-trihydroxyflavone, process of isolation from pterocarpus marsupium and pharmaceutical composition for treatment of diabetes | |
Yesilada et al. | Isolation of anti-ulcerogenic sesquiterpene lactones from Centaurea solstitialis L. ssp. solstitialis through bioassay-guided fractionation procedures in rats | |
EP1495019B1 (en) | Glucopyranoside, process for isolation thereof, pharmaceutical composition containing same and use thereof | |
CN101585825B (en) | Method for preparing amentoflavone | |
KR100460133B1 (en) | Production Method of Catechin with High Antioxidative Activity from The Extracts of Rosa davurica Pall | |
CN110305014B (en) | Cycloneolignanoid enantiomers, preparation and application thereof | |
JP3943082B2 (en) | Substance having molecular structure of 8- (c-β-D-glucopyranosyl) -7,3 ', 4'-trihydroxyflavone, separation method thereof and drug composition using the same | |
EP3705128A1 (en) | Piper laetispicum extract and preparation method therefor and use thereof | |
TWI321052B (en) | Composition for treating cancer cells and preparation method thereof | |
CN101041652B (en) | Separating purified new bisflavone compound from dragon's blood and preparation method thereof | |
KR101316132B1 (en) | A method for separating and purifying cosmetic effective compounds comprising p-coumaric acid from Sasa querlpaertensis Nakai | |
KR100809731B1 (en) | Pharmaceutical composition comprising --secoisolariciresinol | |
KR20060082214A (en) | Novel sargachromanol derivatives, a isolation method thereof, and a composition containing the same showing antioxidant activity | |
KR100377514B1 (en) | Chalcone derivatives, method for preparation thereof and pharmaceutical composition containing the said derivatives | |
USRE37771E1 (en) | Purification of cinnamoyl-C-glycoside chromone | |
CN111675719B (en) | Flavonoid compound and preparation method and application thereof | |
JPH06100584A (en) | New flavonoid glycoside | |
Shakil et al. | Isolation and structure of cordifolin, a novel insecticidal oxygenated chalcone, from the stem of Tinospora cordifolia Miers | |
CN110218208B (en) | Diels-Alder type compound and preparation method and application thereof | |
CN108530505A (en) | A kind of flavonoid glycoside compound and its preparation method and application | |
Gharbo et al. | Phytochemical investigation of Thesium humile Vahl | |
JPH0665278A (en) | New flavonoid glycoside | |
CN116355029A (en) | Dihydrochalcone glycoside compound in traditional Chinese medicine Balanophora japonica and preparation method and application thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20030418 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20061121 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20070208 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20070327 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20070404 |
|
R150 | Certificate of patent or registration of utility model |
Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20100413 Year of fee payment: 3 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20110413 Year of fee payment: 4 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120413 Year of fee payment: 5 |
|
LAPS | Cancellation because of no payment of annual fees |