JP6615455B2 - Method for producing liquiritigenin composition - Google Patents
Method for producing liquiritigenin composition Download PDFInfo
- Publication number
- JP6615455B2 JP6615455B2 JP2014262279A JP2014262279A JP6615455B2 JP 6615455 B2 JP6615455 B2 JP 6615455B2 JP 2014262279 A JP2014262279 A JP 2014262279A JP 2014262279 A JP2014262279 A JP 2014262279A JP 6615455 B2 JP6615455 B2 JP 6615455B2
- Authority
- JP
- Japan
- Prior art keywords
- formula
- liquiritigenin
- represented
- isoliquiritigenin
- reaction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- JBQATDIMBVLPRB-UHFFFAOYSA-N isoliquiritigenin Natural products OC1=CC(O)=CC=C1C1OC2=CC(O)=CC=C2C(=O)C1 JBQATDIMBVLPRB-UHFFFAOYSA-N 0.000 title claims description 56
- FURUXTVZLHCCNA-UHFFFAOYSA-N Liquiritigenin Natural products C1=CC(O)=CC=C1C1OC2=CC(O)=CC=C2C(=O)C1 FURUXTVZLHCCNA-UHFFFAOYSA-N 0.000 title claims description 39
- 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 title claims description 39
- 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 title claims description 38
- 238000004519 manufacturing process Methods 0.000 title claims description 10
- 239000000203 mixture Substances 0.000 title claims description 9
- 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 claims description 18
- 235000008718 isoliquiritigenin Nutrition 0.000 claims description 18
- 238000006243 chemical reaction Methods 0.000 claims description 14
- 150000007524 organic acids Chemical class 0.000 claims description 14
- 239000007864 aqueous solution Substances 0.000 claims description 8
- 125000006239 protecting group Chemical group 0.000 claims description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 24
- 238000000034 method Methods 0.000 description 11
- 235000006200 Glycyrrhiza glabra Nutrition 0.000 description 10
- 235000001453 Glycyrrhiza echinata Nutrition 0.000 description 9
- 235000017382 Glycyrrhiza lepidota Nutrition 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 9
- 229940010454 licorice Drugs 0.000 description 9
- 241000202807 Glycyrrhiza Species 0.000 description 8
- 230000000144 pharmacologic effect Effects 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 6
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- 239000002253 acid Substances 0.000 description 6
- DPZNOMCNRMUKPS-UHFFFAOYSA-N 1,3-Dimethoxybenzene Chemical compound COC1=CC=CC(OC)=C1 DPZNOMCNRMUKPS-UHFFFAOYSA-N 0.000 description 4
- RGHHSNMVTDWUBI-UHFFFAOYSA-N 4-hydroxybenzaldehyde Chemical compound OC1=CC=C(C=O)C=C1 RGHHSNMVTDWUBI-UHFFFAOYSA-N 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- GHMLBKRAJCXXBS-UHFFFAOYSA-N resorcinol Chemical compound OC1=CC=CC(O)=C1 GHMLBKRAJCXXBS-UHFFFAOYSA-N 0.000 description 4
- AFDXODALSZRGIH-QPJJXVBHSA-N (E)-3-(4-methoxyphenyl)prop-2-enoic acid Chemical compound COC1=CC=C(\C=C\C(O)=O)C=C1 AFDXODALSZRGIH-QPJJXVBHSA-N 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 3
- VSCWAEJMTAWNJL-UHFFFAOYSA-K aluminium trichloride Chemical compound Cl[Al](Cl)Cl VSCWAEJMTAWNJL-UHFFFAOYSA-K 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- ZKQFHRVKCYFVCN-UHFFFAOYSA-N ethoxyethane;hexane Chemical compound CCOCC.CCCCCC ZKQFHRVKCYFVCN-UHFFFAOYSA-N 0.000 description 3
- 229940093915 gynecological organic acid Drugs 0.000 description 3
- 235000005985 organic acids Nutrition 0.000 description 3
- AFDXODALSZRGIH-UHFFFAOYSA-N p-coumaric acid methyl ether Natural products COC1=CC=C(C=CC(O)=O)C=C1 AFDXODALSZRGIH-UHFFFAOYSA-N 0.000 description 3
- 239000002243 precursor Substances 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- 231100000820 toxicity test Toxicity 0.000 description 3
- 239000004375 Dextrin Substances 0.000 description 2
- 229920001353 Dextrin Polymers 0.000 description 2
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 2
- 244000303040 Glycyrrhiza glabra Species 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 102000004195 Isomerases Human genes 0.000 description 2
- 108090000769 Isomerases Proteins 0.000 description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 2
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 2
- 235000010323 ascorbic acid Nutrition 0.000 description 2
- 229960005070 ascorbic acid Drugs 0.000 description 2
- 239000011668 ascorbic acid Substances 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 235000019425 dextrin Nutrition 0.000 description 2
- 230000002496 gastric effect Effects 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 208000032839 leukemia Diseases 0.000 description 2
- 201000007270 liver cancer Diseases 0.000 description 2
- 208000014018 liver neoplasm Diseases 0.000 description 2
- 201000005202 lung cancer Diseases 0.000 description 2
- 208000020816 lung neoplasm Diseases 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- CTSLXHKWHWQRSH-UHFFFAOYSA-N oxalyl chloride Chemical compound ClC(=O)C(Cl)=O CTSLXHKWHWQRSH-UHFFFAOYSA-N 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000007363 ring formation reaction Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- DZLOUWYGNATKKZ-VMPITWQZSA-N (e)-3-(4-ethoxyphenyl)prop-2-enoic acid Chemical compound CCOC1=CC=C(\C=C\C(O)=O)C=C1 DZLOUWYGNATKKZ-VMPITWQZSA-N 0.000 description 1
- WBYWAXJHAXSJNI-VOTSOKGWSA-M .beta-Phenylacrylic acid Natural products [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 description 1
- HAEJSGLKJYIYTB-ZZXKWVIFSA-N 4-carboxycinnamic acid Chemical compound OC(=O)\C=C\C1=CC=C(C(O)=O)C=C1 HAEJSGLKJYIYTB-ZZXKWVIFSA-N 0.000 description 1
- DZLOUWYGNATKKZ-UHFFFAOYSA-N 4-ethoxycinnamic acid Natural products CCOC1=CC=C(C=CC(O)=O)C=C1 DZLOUWYGNATKKZ-UHFFFAOYSA-N 0.000 description 1
- 206010007269 Carcinogenicity Diseases 0.000 description 1
- 229920002101 Chitin Polymers 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 1
- 229930183217 Genin Natural products 0.000 description 1
- 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 1
- 235000017443 Hedysarum boreale Nutrition 0.000 description 1
- 235000007858 Hedysarum occidentale Nutrition 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- CSCPPACGZOOCGX-WFGJKAKNSA-N acetone d6 Chemical compound [2H]C([2H])([2H])C(=O)C([2H])([2H])[2H] CSCPPACGZOOCGX-WFGJKAKNSA-N 0.000 description 1
- 229940009456 adriamycin Drugs 0.000 description 1
- 125000003545 alkoxy group Chemical group 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 150000001720 carbohydrates Chemical group 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 231100000260 carcinogenicity Toxicity 0.000 description 1
- 230000007670 carcinogenicity Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- RQFQJYYMBWVMQG-IXDPLRRUSA-N chitotriose Chemical compound O[C@@H]1[C@@H](N)[C@H](O)O[C@H](CO)[C@H]1O[C@H]1[C@H](N)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)N)[C@@H](CO)O1 RQFQJYYMBWVMQG-IXDPLRRUSA-N 0.000 description 1
- 238000011097 chromatography purification Methods 0.000 description 1
- 235000013985 cinnamic acid Nutrition 0.000 description 1
- 229930016911 cinnamic acid Natural products 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- USLKCMBGQFYUFI-UHFFFAOYSA-N dichloromethane;tribromoborane Chemical compound ClCCl.BrB(Br)Br USLKCMBGQFYUFI-UHFFFAOYSA-N 0.000 description 1
- 238000007336 electrophilic substitution reaction Methods 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000005187 foaming Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229960002449 glycine Drugs 0.000 description 1
- 235000013905 glycine and its sodium salt Nutrition 0.000 description 1
- 239000001947 glycyrrhiza glabra rhizome/root Substances 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 230000002140 halogenating effect Effects 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 230000007721 medicinal effect Effects 0.000 description 1
- 229910001507 metal halide Inorganic materials 0.000 description 1
- 150000005309 metal halides Chemical class 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- WBYWAXJHAXSJNI-UHFFFAOYSA-N methyl p-hydroxycinnamate Natural products OC(=O)C=CC1=CC=CC=C1 WBYWAXJHAXSJNI-UHFFFAOYSA-N 0.000 description 1
- 239000012452 mother liquor Substances 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000004366 reverse phase liquid chromatography Methods 0.000 description 1
- 238000006798 ring closing metathesis reaction Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 235000014214 soft drink Nutrition 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Landscapes
- Pyrane Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Description
本発明は甘草素リキリチゲニンの前駆物質イソリキリチゲニンと有機酸を主成分とするリキリチゲニン薬理効果を有する組成物に関する。 The present invention relates to a composition having a pharmacological effect of liquiritigenin, which is mainly composed of a precursor of licorice liquiritigenin, isoliquiritigenin and an organic acid.
甘草素リキリチゲニンは広く薬効が認められる優れた漢方薬であって、一般に甘草から抽出される。しかし、その抽出方法は甘草根を熱抽出した後、加水分解後の糖類部（グルコースやアビオース）を除去し、得られるリキリチゲニンの逆相クロマトグラフによる精製を必要とする。そのため、甘草素の甘草からの抽出はコスト高で、貴重ではあり、1ｇ1万元以上で販売されている。その上甘草の栽培が困難なことから甘草が減少傾向にあって、入手困難に伴い、ますます高騰しつつある。 Licorice liquiritigenin is an excellent herbal medicine with widely recognized medicinal effects, and is generally extracted from licorice. However, the extraction method requires extraction of the liquoritigenin by reverse phase chromatography after removing saccharide parts (glucose and abiose) after hydrolysis after extracting licorice root with heat. Therefore, the extraction of licorice from licorice is expensive and valuable, and it is sold for over 10,000 yuan. In addition, licorice is declining because it is difficult to cultivate licorice.
そこで、甘草素リキリチゲニンを人工的に合成しようとする試みがある。第１の方法は図２に示すように、p−ヒドロキシベンズアルデヒドを出発原料とする方法で、４−カルボキシケイヒ酸を合成し、これを経由して１，３−ジヒドロキシベンゼンを付加してイソリキリチゲニンを得る方法である（非特許文献１）が、収率が低いという欠点がある。そこで、第２の方法として、図３に示すように、p−ヒドロキシベンズアルデヒドを出発原料とし、p位の水酸基をMOM試薬で保護する一方、１，３−ジヒドロキシベンゼンの水酸基をMOM試薬で保護し、両者を付加する方法が提案される（非特許文献２）。しかしながら、実際に検証してみると、ほぼ全段階（収率や生成物がオイル状など）でカラムクロマトによる精製工程が必要となる。しかも、我々の追試実験ではスケールアップした場合のカップリング工程の収率が42%まで低下し、その上最終工程のイソリキリチゲニンからリキリチゲニンへの閉環反応で，原料と目的物が平衡状態にあって，収率が上がらないという結果を得た。また、閉環段階での原料と目的物の分離が，困難で、最終クロマト精製条件でリキリチゲニンはテーリングし，大幅に回収率が低下した。しかもMOM保護試薬の発がん性，さらに保護，脱保護で２段階の余分な工程が必要であり、第２法は実験室レベルでは有効であるものの、量産には適しないとの知見を得た。 There is an attempt to artificially synthesize licorice liquiritigenin. As shown in FIG. 2, the first method is a method in which p-hydroxybenzaldehyde is used as a starting material, and 4-carboxycinnamic acid is synthesized, and 1,3-dihydroxybenzene is added via this to isomerize. Although it is a method for obtaining lithigenin (Non-patent Document 1), there is a disadvantage that the yield is low. Therefore, as shown in FIG. 3, as a second method, p-hydroxybenzaldehyde is used as a starting material, and the hydroxyl group at the p-position is protected with a MOM reagent, while the hydroxyl group of 1,3-dihydroxybenzene is protected with a MOM reagent. A method of adding both is proposed (Non-Patent Document 2). However, when actually verified, purification steps by column chromatography are required in almost all stages (yield, product is oily, etc.). Moreover, in our follow-up experiment, the yield of the coupling process when scaled up decreased to 42%, and in addition, the ring-closing reaction from isoliquiritigenin to liquiritigenin in the final process brought the equilibrium between the raw material and the target product. As a result, the yield was not increased. In addition, it was difficult to separate the raw material and the target product at the ring closure stage, and liquiritigenin tailed under the final chromatographic purification conditions, resulting in a significant reduction in recovery. In addition, two extra steps are required for the carcinogenicity of the MOM protecting reagent, and further protection and deprotection, and the second method was effective at the laboratory level, but it was found that it was not suitable for mass production.
そこで、リキリチゲニンの量産のため、クロマトグラフを使用せず、晶出させる方法を選択し、しかも発がん性のあるMOM保護試薬を使用せず、量産する方法が提案される。さらに人工的にイソリキリチゲニンをリキリチゲニンに変換する場合、平衡反応に伴って収率が向上しないという知見を得た。さらにまた、有機合成したリキリチゲニンは天然物と違って（＋）体と（−）体が共存し、そのまま使用することが懸念され、（＋）体は新たに毒性試験を必要とするとの知見をも得られた。そこで、本発明はリキリチゲニンの前駆物質であるイソリキリチゲニンを有効利用してリキリチゲニンの薬理効果を得る組成物を提供することを目的とする。 Therefore, for mass production of liquiritigenin, a method for mass production without using a chromatograph and selecting a method for crystallization and without using a carcinogenic MOM protecting reagent is proposed. Furthermore, the present inventors have found that when isolyticiligenin is artificially converted to liquiritigenin, the yield does not increase with the equilibrium reaction. Furthermore, the organically synthesized liquiritigenin, unlike natural products, may coexist in the (+) and (-) forms, and may be used as it is, and the (+) form requires a new toxicity test. Was also obtained. Therefore, an object of the present invention is to provide a composition that obtains a pharmacological effect of liquiritigenin by effectively using isoliquiritigenin, which is a precursor of liquiritigenin.
本発明者らは、有機合成的にイソリキリチゲニンをリキリチゲニンに変換すると天然物と違って（＋）体と（−）体が共存するが、人体に有用な有機酸、特にクエン酸を主成分とする有機酸水溶液中で培養すると、（−）体のリキリチゲニンに変換されることに着目し、リキリチゲニンを有機合成せず、その前駆体であるイソリキリチゲニンを合成し、これを酸性の有機酸水溶液、特にクエン酸を主成分とする有機酸水溶液中で培養し、目的のリキリチゲニンに変換することを要旨とするもので、p−アルコキシ桂皮酸を出発原料とし、これにフィーデルクラフト反応を利用してp−アルコキシベンゼンをカップリングさせ、高収率でトリアルコキシイソリキリチゲニンが形成されて晶出し、これを脱保護してイソリキリチゲニンを得、このイソリキリチゲニンを有機酸、好ましくはクエン酸を主成分とする酸性、好ましくはｐＨ３．５から４．５の有機酸水溶液中で培養してリキリチゲニン組成物とし、リキリチゲニン（−）体の薬理効果を得るものである。 The inventors of the present invention organically synthesized isoliqueritigenin into liquiritigenin coexist with (+) and (−) isomers, unlike natural products. However, organic acids useful for the human body, particularly citric acid, are mainly used. Focusing on the fact that when cultured in an organic acid aqueous solution as a component, it is converted to (-) liquiritigenin and does not synthesize liquiritigenin, but synthesizes its precursor, isoliquiritigenin, It is cultivated in an organic acid aqueous solution, particularly an organic acid aqueous solution containing citric acid as a main component, and converted to the target liquiritigenin. The starting material is p-alkoxycinnamic acid, and this is subjected to the Feedel Craft reaction. P-alkoxybenzene is used for coupling, and trialkoxyisolithicigenin is formed and crystallized in a high yield, which is deprotected to obtain isoliquiritigenin. Liquiritigenin is obtained by culturing liquiritigenin in an organic acid aqueous solution having an organic acid, preferably citric acid as a main component, preferably pH 3.5 to 4.5 to obtain a liquiritigenin (-) pharmacological effect. It is.
リキリチゲニン（−）体の薬理効果は特許公報第５６１１３９４号によれば、リキリチゲニン１０μｇ／ｍｌで、人肝臓がん細胞ＳＭＭＣ７７２１に対し９６．０８、人低分化胃線がんＢＯＣ−８２３に対し７３．７６、人早幼粒細胞白血病細胞ＨＬ−６０に対し、６４．４０、人肺がん細胞Ａ５４９に対しやや低いが３５．０６を示し、アドリアマイシンのがん細胞に対する抑制効果を上回っている（同公報第２表および第３表）。本発明によれば、最終的に有機合成による場合のようにリキリチゲニン（＋）体と（−）体が共存しないので、収率の高いイソリキリチゲニンを有効利用することができる。また、人体に有用な有機酸、特にクエン酸を主成分とする有機酸水溶液中で培養するだけで上記薬理効果の高いリキリチゲニン（−）体を得ることができ、有機合成による場合よりも、リキリチゲニン（−）体の収率に優れる。また、有機合成による場合のように、リキリチゲニン（＋）が共存しないから、毒性試験の必要もない。 According to Japanese Patent Publication No. 5611394, the pharmacological effect of liquiritigenin (−) body is 10 μg / ml liquiritigenin, 96.08 for human liver cancer cell SMMC7721, 73. for human poorly differentiated gastric line cancer BOC-823. 76, 64.40 for human early leukemic cell leukemia cell HL-60, 35.06 for human lung cancer cell A549, slightly higher than the inhibitory effect of adriamycin on cancer cells Tables 2 and 3). According to the present invention, since the liquiritigenin (+) form and the (-) form do not coexist as in the case of the final organic synthesis, it is possible to effectively utilize the high yield of isoliquiritigenin. In addition, the liquiritigenin (−) body having a high pharmacological effect can be obtained only by culturing in an organic acid aqueous solution mainly composed of citric acid, which is useful for the human body. (−) Excellent body yield. Moreover, since the liquiritigenin (+) does not coexist as in the case of organic synthesis, there is no need for toxicity tests.
本発明にかかるイソリキリチゲニンは図１で示すようにして製造することができる。
第１にカップリング反応（Ａ）で、式（I）で示されるpーアルコキシ桂皮酸と式（II）で示される１，３−ジアルコキシベンゼンとから式（III）で示されるトリアルコキシイソリキリチゲニンを合成し、保護基を離脱してイソリキリチゲニンを合成する。式（I）、（II）、（III）はその一般式において、Rはメチル、エチルまたはブチルであるアルコキシ基であるのが望ましく、通常入手が容易なRがメチルであるメトキシ基が用いられる。反応は式（I）で示されるp−アルコキシ桂皮酸にハロゲン化剤を添加してハロゲン化し、そこに式（II）で示されるp−アルコキシベンゼンを添加し、次いで、触媒（金属ハロゲン化物、例えば塩化アルミニウム等）存在下に所定の芳香環の水素に求電子置換し、式（IIIａ）で示されるトリアルコキシ桂皮酸が得られる。
The isoliquiritigenin according to the present invention can be produced as shown in FIG.
First, in the coupling reaction (A), a trialkoxyisoloxy represented by the formula (III) from p-alkoxycinnamic acid represented by the formula (I) and 1,3-dialkoxybenzene represented by the formula (II). Lithigenin is synthesized, and isolithicigenin is synthesized by removing the protecting group. In formulas (I), (II) and (III), R is preferably an alkoxy group in which methyl, ethyl or butyl is used, and a methoxy group in which R is methyl is usually available. . The reaction is halogenated by adding a halogenating agent to p-alkoxycinnamic acid represented by formula (I), to which p-alkoxybenzene represented by formula (II) is added, and then a catalyst (metal halide, For example, trialkoxycinnamic acid represented by the formula (IIIa) is obtained by electrophilic substitution of hydrogen of a predetermined aromatic ring in the presence of aluminum chloride or the like.
第２に反応式（B）では、この保護機を脱離すると、式（IV）で示される目的のイソリキリチゲニンが得られる。イソリキリチゲニンは閉環反応（Ｃ）により、式（IV）で示されるイソリキリチゲニンが式（V）で示されるリキリチゲニンに変換されるが、（＋）体と（−)体が得られ、甘草から得られるリキリチゲニンは（−）体であるため、キラル分割により両者を分離する必要があり、製法が複雑となる。また、（＋）体の毒性試験が必要であるので、イソリキリチゲニンのイソメラーゼ酵素を使って、リキリチゲニン（−）体を得るようにする必要がある。本発明ではイソメラーゼを使用することなく、クエン酸を主成分とする有機酸水溶液をｐＨ３．５から４．５に調整し、これにイソリキリチゲニンを添加するとリキリチゲニンの（−）体を得ることができることを見出した。p−アルコキシ桂皮酸としてはp−メトキシ桂皮酸が好ましいが、p−エトキシ桂皮酸、p−ブトキシ桂皮酸を使用しても良い。以下の実施例では、具体的には式（iａ）で示されるp−メトキシ桂皮酸と式（IIa)で示される１，３−ジメトキシベンゼンとから式（IIIa)で示されるトリメトキシイソリキリチゲニンを合成し、保護基を離脱してイソリキリチゲニンを合成する。 Secondly, in the reaction formula (B), when the protective device is removed, the target isoliquiritigenin represented by the formula (IV) is obtained. Isoliquiritigenin is converted to liquiritigenin represented by formula (V) by ring-closure reaction (C), and (+) and (−) isomers are obtained. Since liquiritigenin obtained from licorice is a (−) isomer, it is necessary to separate them by chiral resolution, which complicates the production method. In addition, since a toxicity test of the (+) form is necessary, it is necessary to obtain a liquiritigenin (-) form using an isomerase enzyme of isoliquiritigenin. In the present invention, without using isomerase, an organic acid aqueous solution containing citric acid as a main component is adjusted to pH 3.5 to 4.5, and by adding isoliquiritigenin to this, a (−) isomer of liquiritigenin is obtained. I found out that I can. As p-alkoxycinnamic acid, p-methoxycinnamic acid is preferable, but p-ethoxycinnamic acid and p-butoxy cinnamic acid may be used. In the following examples, specifically, trimethoxyisolithilic acid represented by the formula (IIIa) from p-methoxycinnamic acid represented by the formula (ia) and 1,3-dimethoxybenzene represented by the formula (IIa). Genine is synthesized, and the protective group is removed to synthesize isoliquiritigenin.
イソリキリチゲニンを培養する有機酸水溶液はクエン酸を主成分として構成される。クエン酸以外の有機酸としてはアスコルビン酸、アミノ酸、酢酸等の種々の有機酸を使用することができる。また、ｐＨ３．５から４．５とすることが重要であり、培養時間は短くてもよいが、１０時間以上有機酸中で培養するのがよく、イソリキリチゲニンのリキリチゲニンへの変換を阻害しない限り、その他の付形剤を添加してもよい。 An organic acid aqueous solution for culturing isoliquiritigenin is composed mainly of citric acid. As organic acids other than citric acid, various organic acids such as ascorbic acid, amino acids, and acetic acid can be used. Further, it is important to adjust the pH to 3.5 to 4.5, and the culture time may be short, but it is preferable to culture in an organic acid for 10 hours or more and inhibit the conversion of isoliquiritigenin to liquiritigenin. Other excipients may be added as long as they are not.
以下、本発明の好ましい具体例を実施例に基づいて説明する。
（実施例１）
Hereinafter, preferred specific examples of the present invention will be described based on examples.
Example 1
式(Ia) 式（IIIa)
4-メトキシケイヒ酸 式(Ia）10ｇをジメチルホルムアミド0.25 mLを含む無水塩化メチレン50 mLに溶解し，室温にてオキザリルクロリド9.6 mLを発泡に注意しながら10分かけて滴下した。そのまま，室温にて2時間撹拌の後，溶媒を減圧下で除去した。得られた残渣に，1，3-ジメトキシベンゼン7.4 mL（式IIa）と無水エーテル200 mLを加え，氷浴中で，粉末にした触媒無水三塩化アルミニウム22.4 gを15分かけてゆっくりと加えた。そのまま室温にて一晩放置した後，内容物を氷上(500 g)にあけ，６M 塩酸を10 mL加え，氷が溶解した後，酢酸エチル(300 mL)で4回抽出した。抽出液を無水硫酸ナトリウムで乾燥後，減圧下で濃縮し，残渣をエーテル-ヘキサン混液で結晶化することにより，14.2 gの目的物(式IIIa)の結晶を得た。収率85% 1H-NMR (CDCl3) δ 7.73 (1H, d, J=8.1 Hz), 7.64 (1H, d, J=15.1 Hz), 7.54 (2H, d, J=7.7 Hz), 7.38 (1H, d, J=15.1 Hz), 6.90 (2H, d, J=7.7 Hz), 6.55 (1H, brd, J=8.1 Hz), 6.49 (1H, brs), 3.89 (3H, s), 3.85 (3H, s), 3.83 (3H, s).
Formula (Ia) Formula (IIIa)
10 g of 4-methoxycinnamic acid formula (Ia) was dissolved in 50 mL of anhydrous methylene chloride containing 0.25 mL of dimethylformamide, and 9.6 mL of oxalyl chloride was added dropwise at room temperature over 10 minutes while paying attention to foaming. After stirring for 2 hours at room temperature, the solvent was removed under reduced pressure. To the obtained residue, 7.4 mL of 1,3-dimethoxybenzene (formula IIa) and 200 mL of anhydrous ether were added, and 22.4 g of powdered catalyst anhydrous aluminum trichloride was slowly added over 15 minutes in an ice bath. . The mixture was allowed to stand at room temperature overnight, then the contents were poured on ice (500 g), 10 mL of 6M hydrochloric acid was added, and the ice was dissolved, followed by extraction four times with ethyl acetate (300 mL). The extract was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was crystallized from an ether-hexane mixture to obtain 14.2 g of the desired product (formula IIIa). Yield 85% 1 H-NMR (CDCl 3 ) δ 7.73 (1H, d, J = 8.1 Hz), 7.64 (1H, d, J = 15.1 Hz), 7.54 (2H, d, J = 7.7 Hz), 7.38 (1H, d, J = 15.1 Hz), 6.90 (2H, d, J = 7.7 Hz), 6.55 (1H, brd, J = 8.1 Hz), 6.49 (1H, brs), 3.89 (3H, s), 3.85 (3H, s), 3.83 (3H, s).
式（IIIa） 式（III)
上記生成物3 gを塩化メチレン60 mLに溶解し0 ℃にて1 M BBr3塩化メチレン溶液を滴下した。室温まで昇温し，そのまま2日間撹拌した。セニエット塩34 gを含む氷冷水700 mLとメタノール350 mLを加え，室温で一晩撹拌した。得られた黄色溶液を酢酸エチルで2回抽出し，1 Mセニエット塩-飽和食塩水で洗浄後，無水硫酸ナトリウムで乾燥，濃縮した。残渣をエーテル-ヘキサンで結晶化し，目的物1.95 g（式IV）を得た。またその母液を濃縮後再度エーテル-ヘキサンで結晶化することにより0.59 gの目的物第２晶を得た。合計収率98%1H-NMR (acetone-d6) δ 13.5 (1H, s), 8.10 (1H, d, J=8.3 Hz), 7.82 (1H, d, J=15.4 Hz), 7.74 (1H, d, J=15.4 Hz), 7.72 (2H, d, J=8.2 Hz), 6.90 (2H, d, J=8.2 Hz), 6.44 (1H, dd, J=8.3 and 1.7 Hz), 6.34 (1H, d, J=1.7 Hz).
（実施例２）
Formula (IIIa) Formula (III)
3 g of the above product was dissolved in 60 mL of methylene chloride, and 1 M BBr 3 methylene chloride solution was added dropwise at 0 ° C. The mixture was warmed to room temperature and stirred for 2 days. Ice-cooled water (700 mL) containing 34 g of Sennet salt and methanol (350 mL) were added, and the mixture was stirred overnight at room temperature. The resulting yellow solution was extracted twice with ethyl acetate, washed with 1 M seniette-saturated brine, dried over anhydrous sodium sulfate and concentrated. The residue was crystallized from ether-hexane to obtain 1.95 g (formula IV) of the desired product. The mother liquor was concentrated and recrystallized from ether-hexane to obtain 0.59 g of the second crystal of the desired product. Total yield 98% 1 H-NMR (acetone-d 6 ) δ 13.5 (1H, s), 8.10 (1H, d, J = 8.3 Hz), 7.82 (1H, d, J = 15.4 Hz), 7.74 (1H , d, J = 15.4 Hz), 7.72 (2H, d, J = 8.2 Hz), 6.90 (2H, d, J = 8.2 Hz), 6.44 (1H, dd, J = 8.3 and 1.7 Hz), 6.34 (1H , d, J = 1.7 Hz).
(Example 2)
式（IV） 式（V）
原料200 mg（式IV）をｐＨ４前後に調整したクエン酸水溶液に加え，撹拌し，よく分散した後に，１昼夜常温で放置した。その懸濁液を，常法で結晶化することにより粗結晶を得る（式V）。その分析値は以下の通りであり、リキリチゲニンへ変換されている。
1H-NMR (DMSO-d6) δ 9.65 (1H, brs), 7.58 (1H, m), 7.27 (2H, m), 6.74 (2H, m), 6.45 (1H, m), 6.28 (1H, m), 5.39 (1H, brd, J=11.6 Hz), 3.65 (1H, brt, J=15.0 Hz), 2.58 (1H, brd, J=15.7 Hz).
（実施例３）
Formula (IV) Formula (V)
200 mg (formula IV) of the raw material was added to an aqueous citric acid solution adjusted to around
1H-NMR (DMSO-d 6 ) δ 9.65 (1H, brs), 7.58 (1H, m), 7.27 (2H, m), 6.74 (2H, m), 6.45 (1H, m), 6.28 (1H, m ), 5.39 (1H, brd, J = 11.6 Hz), 3.65 (1H, brt, J = 15.0 Hz), 2.58 (1H, brd, J = 15.7 Hz).
(Example 3)
難消化デキストリン、Ｎ−アセチルグルコサミン、デキストリン、キチンオリゴ糖、キトサンオリゴ糖、乳酸、アスコルビン酸（ビタミンＣ）を主成分とする清涼飲料水５０ｍｌ「長寿挑戦」：株式会社国際メディカル研究所販売は、ｐＨ３．９である。そこで、リキリチゲニンとしてイソリキリチゲニンを下記特許第５６１１３９４号明細書に記載の表２「リキリチゲニンのがん細胞に対する抑制作用」に示す用量（マウスに対する有効量）の約１００〜２０００倍のイソリキリチゲニンを添加し、免疫増強のためのサプリメントを形成した。
特許公報第５６１１３９４号の表２によれば、リキリチゲニン（−）体の薬理効果はリキリチゲニン１０μｇ／ｍｌで、人肝臓がん細胞ＳＭＭＣ７７２１に対し９６．０８、人低分化胃線がんＢＯＣ−８２３に対し７３．７６、人早幼粒細胞白血病細胞ＨＬ−６０に対し、６４．４０、人肺がん細胞Ａ５４９に対しやや低いが３５．０６を示すことが明らかにされている。
50ml “Longevity Challenge” of soft drinks consisting mainly of indigestible dextrin, N-acetylglucosamine, dextrin, chitin oligosaccharide, chitosan oligosaccharide, lactic acid, ascorbic acid (vitamin C): The pH is 3.9. Accordingly, as the liquiritigenin, isoliquiritigenin is about 100 to 2000 times the dose (effective amount for mice) shown in Table 2 “Inhibitory action of liquiritigenin on cancer cells” described in the following patent No. 5611394. Genin was added to form a supplement for immune enhancement.
According to Table 2 of Japanese Patent Publication No. 5611394, the pharmacological effect of liquiritigenin (−) is 10 μg / ml liquiritigenin, which is 96.08 against human liver cancer cell SMMC7721, and human poorly differentiated gastric line cancer BOC-823. On the other hand, 73.76, 64.40 for human early granulocyte leukemia cell HL-60, and 35.06 for human lung cancer cell A549 were shown to be slightly lower.
Claims (2)
Priority Applications (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2014262279A JP6615455B2 (en) | 2014-12-25 | 2014-12-25 | Method for producing liquiritigenin composition |
| PCT/JP2015/069021 WO2016002848A1 (en) | 2014-07-02 | 2015-07-01 | Process for producing liquiritigenin precursor |
| US15/322,582 US10023552B2 (en) | 2014-07-02 | 2015-07-01 | Method of preparing a liquiritigenin precursor |
| CN201580036037.2A CN106536468B (en) | 2014-07-02 | 2015-07-01 | Preparation method of liquiritigenin precursor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2014262279A JP6615455B2 (en) | 2014-12-25 | 2014-12-25 | Method for producing liquiritigenin composition |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2016121101A JP2016121101A (en) | 2016-07-07 |
| JP6615455B2 true JP6615455B2 (en) | 2019-12-04 |
Family
ID=56327099
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2014262279A Active JP6615455B2 (en) | 2014-07-02 | 2014-12-25 | Method for producing liquiritigenin composition |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP6615455B2 (en) |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1175425A2 (en) * | 1999-01-21 | 2002-01-30 | The Board Of Regents, The University Of Texas System | Inhibitors of intestinal apical membrane na/phosphate co-transportation |
-
2014
- 2014-12-25 JP JP2014262279A patent/JP6615455B2/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| JP2016121101A (en) | 2016-07-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US8084430B2 (en) | ENT-kaurene diterpene compound and its derivatives, their preparation and their use | |
| JP5021549B2 (en) | Composition for the treatment of cancer cells and method of synthesis thereof | |
| Ecer et al. | Efficient and shortcut syntheses of some novel eight-membered ring cyclitols starting from cycloocta-1, 3-diene | |
| CN104447919B (en) | A kind of process for purification of 20,23 dipiperidino, 5 O mycaminose Ji Taile lactone bulk drug | |
| HUE033085T2 (en) | Process for producing a solid form of abiraterone acetate | |
| JP2016502539A (en) | Composition for prevention or treatment of heart disease | |
| KR20100120973A (en) | Method for preparating ascorbic acid derivatives | |
| JP6615455B2 (en) | Method for producing liquiritigenin composition | |
| CN103360411A (en) | Everolimus crystallization purification method | |
| CN106916151A (en) | A kind of preparation method of Lurasidone HCl | |
| WO2016002848A1 (en) | Process for producing liquiritigenin precursor | |
| Ji et al. | A new polyacetylene glycoside from the rhizomes of Atractylodes lancea | |
| CN103992263B (en) | A kind of purification process of E2020 | |
| JP6358877B2 (en) | Method for producing liquiritigenin precursor | |
| TWI703120B (en) | Method for producing liquiritigenin precursor | |
| CN103340881B (en) | The application in neuroprotective of a kind of oligosaccharide compound | |
| Villedieu et al. | A simple and selective method for the O-AcCl removal using sodium borohydride | |
| CN106810563B (en) | Furans card mountain alkane forskolin and its pharmaceutical composition and its application in pharmacy | |
| KR101749132B1 (en) | Composition for Preventing or Treating Inflammatory Diseases Comprising Tanzawaic Acid | |
| CN103012175A (en) | Anti-tumor nitrogenous substituent curcumin analogue, salt of curcumin analogue and preparation method | |
| CN109206461A (en) | The salt of optically active xylose uronic acid derivative and its prepare purification process | |
| CN109251179B (en) | 1,2, 4-triazine gastric cancer cell growth inhibitor and preparation method and application thereof | |
| TWI804600B (en) | METHOD FOR SYNTHESIZING 3-PHENYL-2,8-DIHYDROPYRANO[2,3-ƒ]CHROMENE DERIVATIVES | |
| KR20180079967A (en) | A new synthesis method of (+)-Decursinol | |
| CN110330543A (en) | 3- acetyl group -5- hydroxyl-B- norcholesterol -6-(N- methyl) contracting ammonia sulphur hydrazone, preparation method and its usage |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20171225 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20181128 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20181207 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20190527 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20190725 |
|
| 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: 20191009 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20191106 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 6615455 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |