CN103641884B - A kind of method extracting Triptolide 12,13-chlorohydrin in trypterygine leading portion medicinal extract - Google Patents
A kind of method extracting Triptolide 12,13-chlorohydrin in trypterygine leading portion medicinal extract Download PDFInfo
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- CN103641884B CN103641884B CN201310604804.8A CN201310604804A CN103641884B CN 103641884 B CN103641884 B CN 103641884B CN 201310604804 A CN201310604804 A CN 201310604804A CN 103641884 B CN103641884 B CN 103641884B
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Abstract
The invention discloses a kind of method extracting Triptolide 12,13-chlorohydrin in trypterygine leading portion medicinal extract.At present to Tripterysium Glucosides or directly carry out chromatographic separation with the extract after extraction using alcohol trypterygine raw material, technique is the method adopting repeatedly chromatography, and through rotating thin layer chromato-graphy, the Triptolide 12,13-chlorohydrin amount finally obtained is little, can not be used for large production at all.First the present invention uses hydrophilic solvent lower alcohol/ketone to extract tripterygium glycosides leading portion medicinal extract, lipophilic contaminant in most of leading portion medicinal extract is separated, then chloroform extraction is used, other non-targeted product separations of less solubleness will be had in lower alcohol/ketone solution, then chromatography separation and purification, finally uses Gossypol recrystallized from chloroform.Separation of the present invention, purifying process are simple, and be applicable to industrialized production, obtained Triptolide 12,13-chlorohydrin can be used for developing new drug.
Description
Technical field
The present invention relates to the method preparing natural chemicals in plant extract medicinal extract, specifically a kind of method extracting single component Triptolide 12,13-chlorohydrin from trypterygine leading portion medicinal extract.
Background technology
Triptolide 12,13-chlorohydrin (Tripchlorolide) is from Celastraceae tripterygium plant, be separated the Diterpenoid epoxide lactone compound obtained, pharmacological testing shows, it has stronger anti-inflammatory, immunosuppression and Male antifertilityagent, its tire be current clinical application " tripterygium glycosides " preparation 100-200 doubly, it is one of the chief active monomer of trypterygine display anti-inflammatory, immunosuppression and Male antifertilityagent, and be proved without mutagenesis, but this compound is extremely low by former plant extract yield.
China Patent No. 89105433.2 discloses preparation method and the antifertility purposes of tripchlorolide, be characterized in Tripterysium Glucosides or directly carry out chromatographic separation with the extract after extraction using alcohol trypterygine raw material, technique is the method adopting repeatedly chromatography, and through rotating thin layer chromato-graphy, the Triptolide 12,13-chlorohydrin amount finally obtained is little, can not be used for large production at all.
China Patent No. 200810071485.8 discloses the novelty teabag of tripchlorolide, T 4, provides the A β of tripchlorolide, T 4 in the mediation for the treatment of microglia
1-42with the novelty teabag of the nerve immunity inflammatory diseases caused by the neurotoxic effect of LPS, thus be applied to nerve immunity inflammatory diseases for tripchlorolide, T 4, especially the control of alzheimer's disease provides directly strong experimental basis and theoretical basis, existing very strong science and novelty, have again very high development and application values.
Summary of the invention
Technical problem to be solved by this invention produces the trypterygine monomer effective constituent extracting a kind of high-efficiency low-toxicity dead meal, to provide highly purified Triptolide 12,13-chlorohydrin, for developing new trypterygine monomer formulation from tripterygium glycosides.
For this reason, the present invention adopts following technical scheme: a kind of method extracting Triptolide 12,13-chlorohydrin in trypterygine leading portion medicinal extract, its step is as follows: 1) transform and extract: weighed by the leading portion concentrated extract given up in tripterygium glycosides chromatographic separation process, add solvent lower ketones or the lower alcohol of 3-8 times amount by weight, add the hydrochloric acid of quantity of solvent 2-5% again, the mass percent of hydrochloric acid used is 30-40%, heating and refluxing extraction 2-6 hour, the water with solvent equimultiple is added in the solution obtained, stir, then cooling is left standstill, layering;
2) concentrated, extraction: by step 1) the upper strata red solution of gained distills, recycling design, obtains concentrated extracting solution, then in concentrated extracting solution, adds chloroform extraction isopyknic with it three times, extraction liquid is reclaimed decompressing and extracting after trichloromethane, obtain Powdered crude extract;
3) purifying: described purifying fills silicagel column after being dissolved by Powdered crude extract trichloromethane, carries out chromatographic separation purifying, collect corresponding elutriant, obtain Powdered extract, finally use Gossypol recrystallized from chloroform after desolventizing drying.
The present invention utilize the triptolide that contains in tripterygium glycosides leading portion waste liquid in acid condition can with lower alcohol/reactive ketone, change into the feature of the Triptolide 12,13-chlorohydrin of high-efficiency low-toxicity, first hydrophilic solvent lower alcohol/ketone is used to extract, lipophilic contaminant in most of leading portion medicinal extract is separated, then chloroform extraction is used, other non-targeted product separations of less solubleness will be had in lower alcohol/ketone solution, in the crude extract of at this moment gained, Triptolide 12,13-chlorohydrin content is improved largely (from 100,000/several bring up to some thousandths of), greatly can simplify later separation and purification process, recrystallization can be carried out after one step chromatography, more easily carry out large production operation.
Further, described lower ketones is acetone, and described lower alcohol is methyl alcohol or ethanol.
Further, the order number of silica gel for chromatography is 200-300 order, and before dress post, the moisture of silica gel is adjusted to 8-10%, eluent added in its chromatography process is the mixed solvent of chloroform-ethanol, and its volume ratio is 99:1-95:5.Select above-mentioned chromatography processing condition, enrichment Triptolide 12,13-chlorohydrin, make its content bring up to more than 10% from 2 ‰.
Further, in the Powdered crude extract of gained in trypterygine chlorine the content of alcohol at 2-10 ‰; In gained extract in trypterygine chlorine the content of alcohol at 10-40%.
The present invention takes full advantage of the waste in tripterygium glycosides leaching process, is conducive to the medicine resource making full use of trypterygine, and obtains the trypterygine medicine of high-efficiency low-toxicity; The method that the present invention adopts makes Triptolide 12,13-chlorohydrin content in the crude extract of gained be improved largely, and enormously simplify later separation and purification process, can carry out recrystallization, more easily carry out large production operation after a step chromatography.Separation of the present invention, purifying process are simple, and be applicable to industrialized production, obtained Triptolide 12,13-chlorohydrin can be used for developing new drug.
Below in conjunction with embodiment, the present invention is described further, but be not limited to the scope of the present embodiment.
Embodiment
embodiment 1
Get trypterygine leading portion concentrated extract 4100g, add acetone 4500ml and hydrochloric acid 400ml, put in 10000ml round-bottomed flask, put into water-bath 80 DEG C of reflux, in time starting to reflux, react 2 hours, take out, add purified water 4500ml, stir with sticking plaster, hold over night; Within second day, separate upper strata acetone soln, with filter paper filtering, filtrate is adjusted to 80 DEG C after putting first 65 DEG C of recovery acetone in water-bath (every 2000ml recovery 600ml) and is recycled to about dope about 1000ml, incline and, filtrate adds chloroform extraction three times, and extraction liquid decompressing and extracting after reclaiming chloroform obtains conversion product powder 241.986g.Silicagel column on conversion product 1:10, ethanol-chloroform gradient elution, collect T4, drain, dry product adds Gossypol recrystallized from chloroform and obtains Triptolide 12,13-chlorohydrin crystallization 160mg.
embodiment 2
Get trypterygine leading portion concentrated extract 30Kg and add ethanol 90Kg, hydrochloric acid 1800ml, in enamel reactor, steam heating refluxes 6 hours, puts to plastics storage bucket, adds water 90Kg, open stirrer and stir 15 minutes, hold over night; Second innocent suction goes out upper solution, in enamel reactor, reclaim ethanol, be concentrated into about 1/3 amount time release, filtrate adds chloroform extraction three times, and extraction liquid obtains crude extract powder 7.215Kg(yield 24.05% through reclaiming decompressing and extracting after chloroform).Crude extract powder dress stainless steel huge pillar (30 × 500cm) is carried out silica gel column chromatography separation, obtains extract powder 1343.3g.Extract powder Gossypol recrystallized from chloroform is obtained white crystals 161.2g twice, and surveying content is 98.5% Triptolide 12,13-chlorohydrin.
embodiment 3
Get trypterygine leading portion concentrated extract 100Kg and add methyl alcohol 500Kg, hydrochloric acid 20L, in enamel reactor, steam heating refluxes 6 hours, puts to plastics storage bucket, adds water 500Kg, open stirrer and stir 15 minutes, hold over night; Second innocent suction goes out upper solution, in enamel reactor, reclaim methyl alcohol, be concentrated into about 1/3 amount time release, filtrate adds chloroform extraction three times, and extraction liquid obtains crude extract powder 20.14Kg(yield 20.14% through reclaiming decompressing and extracting after chloroform).Crude extract powder dress stainless steel huge pillar (30 × 500cm) is carried out silica gel column chromatography separation, obtains extract powder 2348.6g.Extract powder Gossypol recrystallized from chloroform is obtained white crystals 422.75g twice, and surveying content is 99.01% Triptolide 12,13-chlorohydrin.
In above-described embodiment, the high performance liquid phase detection method of Triptolide 12,13-chlorohydrin is: high performance liquid chromatograph Agilent1260; Moving phase: Methanol-water (50:50); Chromatographic column: C
18post (4.6 × 250mm); Determined wavelength selects 219nm; Flow velocity is: 1.0ml/min, ambient operation.
TLC method in chromatography process: silica gel g thin-layer plate 10 × 20cm; Developping agent: hexanaphthene-acetone (5:3), ethanol-chloroform (5:95); Developer is: Keddes reagent.
Claims (3)
1. one kind is extracted the method for Triptolide 12,13-chlorohydrin in trypterygine leading portion medicinal extract, its step is as follows: 1) transform and extract: weighed by the leading portion concentrated extract given up in tripterygium glycosides chromatographic separation process, add solvent lower ketones or the lower alcohol of 3-8 times amount by weight, add the hydrochloric acid of quantity of solvent 2-5% again, the mass percent of hydrochloric acid used is 30-40%, heating and refluxing extraction 2-6 hour, the water with solvent equimultiple is added in the solution obtained, stir, then cooling is left standstill, layering;
2) concentrated, extraction: by step 1) the upper strata red solution of gained distills, recycling design, obtains concentrated extracting solution, then in concentrated extracting solution, adds chloroform extraction isopyknic with it three times, extraction liquid is reclaimed decompressing and extracting after trichloromethane, obtain Powdered crude extract;
3) purifying: described purifying fills silicagel column after being dissolved by Powdered crude extract trichloromethane, carries out chromatographic separation purifying, collect corresponding elutriant, obtain Powdered extract, finally use Gossypol recrystallized from chloroform after desolventizing drying;
Described lower ketones is acetone, and described lower alcohol is methyl alcohol or ethanol.
2. the method extracting Triptolide 12,13-chlorohydrin in trypterygine leading portion medicinal extract according to claim 1, it is characterized in that, the order number of silica gel for chromatography is 200-300 order, before dress post, the moisture of silica gel is adjusted to 8-10%, eluent added in its chromatography process is the mixed solvent of chloroform-ethanol, and its volume ratio is 99:1-95:5.
3. the method extracting Triptolide 12,13-chlorohydrin in trypterygine leading portion medicinal extract according to claim 1, is characterized in that, in the Powdered crude extract of gained, the content of Triptolide 12,13-chlorohydrin is at 2-10 ‰; In gained extract, the content of Triptolide 12,13-chlorohydrin is at 10-40%.
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| CN111297934A (en) * | 2019-12-18 | 2020-06-19 | 湖南千金协力药业有限公司 | Tripterygium glycosides extraction method |
| CN114716503B (en) * | 2022-03-23 | 2024-03-12 | 广东省中医院（广州中医药大学第二附属医院、广州中医药大学第二临床医学院、广东省中医药科学院） | Preparation method of tripterygium wilfordii chlorolactone alcohol |
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| CN1052860A (en) * | 1989-12-22 | 1991-07-10 | 中国医学科学院皮肤病研究所 | The preparation method of tripchlorolide and antifertility purposes |
| CN1634493A (en) * | 2004-12-02 | 2005-07-06 | 杨更亮 | Chinese traditional medicine Tripterygium wilfordii and improving technique for attenuation and synergy of its extract |
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| CN101991633A (en) * | 2009-10-10 | 2011-03-30 | 上海复旦复华药业有限公司 | Method for extracting tripterygium glycosides, and product and inclusion compound and medicinal composition thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US20040018260A1 (en) * | 2002-06-19 | 2004-01-29 | Novemed Group Limited | Novel botanical extract of Tripterygium Wilfordii Hook F. |
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| CN1052860A (en) * | 1989-12-22 | 1991-07-10 | 中国医学科学院皮肤病研究所 | The preparation method of tripchlorolide and antifertility purposes |
| CN1634493A (en) * | 2004-12-02 | 2005-07-06 | 杨更亮 | Chinese traditional medicine Tripterygium wilfordii and improving technique for attenuation and synergy of its extract |
| CN100998631A (en) * | 2006-01-13 | 2007-07-18 | 杨更亮 | Medicine form of low toxin thunder godvine extract by using new processing method |
| CN101991633A (en) * | 2009-10-10 | 2011-03-30 | 上海复旦复华药业有限公司 | Method for extracting tripterygium glycosides, and product and inclusion compound and medicinal composition thereof |
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| Tripterygium wilfordii bioactive compounds as anticancer and anti-inflammatory agents;Wong KF，et al.;《Clinical and Experimental Pharmacology and Physiology》;20120222;第39卷(第3期);第311-320页 * |
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Address after: 312560 Zhejiang city of Shaoxing province Xinchang Ru'ao Town Road No. 2 National Road Patentee after: Zhejiang deende pharmaceutical Limited by Share Ltd Address before: 312560 Zhejiang city of Shaoxing province Xinchang Ru'ao Town Road No. 2 National Road Patentee before: Zhejiang De'ende Pharmaceutical Co., Ltd. |