CN1634493A - Chinese traditional medicine Tripterygium wilfordii and improving technique for attenuation and synergy of its extract - Google Patents
Chinese traditional medicine Tripterygium wilfordii and improving technique for attenuation and synergy of its extract Download PDFInfo
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- CN1634493A CN1634493A CN 200410096718 CN200410096718A CN1634493A CN 1634493 A CN1634493 A CN 1634493A CN 200410096718 CN200410096718 CN 200410096718 CN 200410096718 A CN200410096718 A CN 200410096718A CN 1634493 A CN1634493 A CN 1634493A
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Abstract
The invention discloses a modifying process of attenuation and synergy for Tripterygium wilfordii and its extract, wherein the procedure (1) comprises immersing Tripterygium wilfordii with acid solution, conversing Triptolidenol Triptonide into Tripchlo-rolide (II). While the procedure (2) comprises preparing concrete from Tripterygium wilfordii, then purifying Tripterygium wilfordii or its concrete, acid treating or immersing with acid solution.
Description
Technical field
The present invention relates to natural drug extraction, purification and authentication method, particularly the modified technique of Chinese medicine Radix Tripterygii Wilfordii and extract thereof.
Background technology
Radix Tripterygii Wilfordii general reference Celastraceae Radix Tripterygii Wilfordii plant, in homemade four kinds, wherein have three kinds to be used as medicine, promptly Radix Tripterygii Wilfordii (Triptergium wilfordii Hook.f.) originates in provinces and regions such as Zhejiang, Anhui, Hunan, Fujian, mainly in area, the middle and lower reach of Yangtze River; Tripterygium hypoglaucum (T.hypoglaucum Hutch) originates in the Yangtze river basin and southwest; Tripterygium regelii Spragus et Takeda (T.regelii Spragnc et Takcda) claims black climing again, mainly originates in northeast and Japan.Radix Tripterygii Wilfordii is recorded the earliest in Shennong's Herbal, clinical for many years treatment rheumatoid arthritis, chronic nephritis, hepatitis, thrombocytopenic purpura and the various dermatosis of being widely used in.The research of the aspect such as repelling at present after antifertility, antitumor, anti-organ transplantation in the body has also obtained bigger progress, is a kind of rising medicine.Since reported first in 1936 gets terpenoid pigment trypterygine from Radix Tripterygii Wilfordii root extraction separation, from tripterygium plant, isolated more than 100 kind of composition, wherein Diterpenes and alkaloids are main active component.
The Radix Tripterygii Wilfordii Herb is all poisonous, if improper use, period in a medicine often takes place with toxicity.Zoopery shows that the most responsive target organ of Radix Tripterygii Wilfordii toxicity is a gastronintestinal system, and hemopoietic system and reproductive system are the most common with digestive tract reaction.Epoxy diterpene-kind compound Radix Tripterygii Wilfordii lactone alcohol (I) also be main toxic component, and effective dose is almost suitable with toxicity dose for the main effective ingredient of treatment rheumatoid arthritis, and this hinders the further popularization of Chinese medicine Radix Tripterygii Wilfordii aspect clinical practice greatly.The important epoxy diterpene-kind compound of another one in the Radix Tripterygii Wilfordii is tripchlorolide (II), the pharmacologically active The selection result shows, II has antiinflammatory, immunosuppressant and the antifertility activity of I, and tiring is about 100～200 times of I, and has been proved no mutagenic action.Chemistry is related based on having between Compound I I and the I, utilizes the higher I of relative amount to realize that II's is semi-synthetic in laboratory.But the content of Compound I in former plant is not high yet, and this has just limited the application of semisynthesis greatly, and it is extremely low by former plant extract yield to add Compound I I, and its raw material sources have become a vital problem.
Experimentation shows that the long-time decoction of slow fire can reduce the toxicity of Radix Tripterygii Wilfordii effectively, but destroyed because of many effective ingredient, its curative effect also descends to some extent.Therefore can utilize method of modifying that the I in the medical material is carried out structure of modification, to obtain the Radix Tripterygii Wilfordii medicine of high-efficiency low-toxicity.Domesticly still belong to vacancy in the major technique aspect the modification of Radix Tripterygii Wilfordii attenuation synergistic.
Summary of the invention
The objective of the invention is to remedy the deficiencies in the prior art part, and the modified technique of the Chinese medicine Radix Tripterygii Wilfordii extract that a kind of cost is low, toxicity is little is provided.
This technological principle is: adopt acid system, handle Radix Tripterygii Wilfordii (herb or part) or from Radix Tripterygii Wilfordii, extract the extractum that makes, characteristics according to the physicochemical property of main activity, toxic component, make its main active, also be that main toxic component structure changes, the toxicity of Radix Tripterygii Wilfordii reduces like this, promotes its industrialization as drug developments such as antiinflammatory, antitumor.
The object of the invention can realize by following measure: at first adopt Radix Tripterygii Wilfordii (herb or part) or the extractum that makes from Radix Tripterygii Wilfordii is extracted, with the extracting of middle polarity organic solvent, extract is crossed the neutral alumina post earlier and is carried out preliminary purification, remove pigment and little polar impurity part, with the active part of collecting, handle with acid system.Extractum and middle polarity extractive with organic solvent thereof that last acid system is directly handled Radix Tripterygii Wilfordii (herb or part) or made from Radix Tripterygii Wilfordii is extracted prove that through pharmacological evaluation its toxicity reduces, and curative effect is constant.
Concrete modified technique of the present invention carries out according to the following steps:
The first step: total composition extracts in the Radix Tripterygii Wilfordii (herb or part), gets extractum
With a certain amount of Radix et Rhizoma Tripterygii, with polar solvent or middle polarity organic solvent extraction, the amount of Radix Tripterygii Wilfordii in solvent is 0.001--1.8g/ml, and the sucking filtration extracting solution reclaims solvent, gets extractum.
Second step: Radix Tripterygii Wilfordii (herb or part) or by the purification of the extractum of first step gained Radix Tripterygii Wilfordii
With Radix Tripterygii Wilfordii (herb or part) or by the extractum of first step gained Radix Tripterygii Wilfordii, use the middle polarity organic solvent extraction respectively, the amount of Radix Tripterygii Wilfordii in solvent is 0.001-1.2g/ml, after extract concentrates, the chromatographic column separation and purification gets, and gets the higher diterpene-kind compound of purity.
The 3rd step: the Radix Tripterygii Wilfordii behind the purification (herb or part) or by the acid treatment of the extractum of first step gained Radix Tripterygii Wilfordii
Radix Tripterygii Wilfordii behind the purification (herb or part) or by the extractum of first step gained Radix Tripterygii Wilfordii is handled with hydrochloric acid or hydrochloric acid solution respectively, and wherein the content of extractum is 0.001--2.0g/ml.
1), Radix Tripterygii Wilfordii or the 4th step:, carry out acid solution immersion, reuse middle polarity organic solvent extraction, washing respectively, must concentrate modifier by the extractum of first step gained Radix Tripterygii Wilfordii;
Get a certain amount of Radix Tripterygii Wilfordii or, soak with hydrochloric acid or hydrochloric acid solution respectively, handle down, make Radix Tripterygii Wilfordii lactone alcohol major part wherein be converted into tripchlorolide at 0-40 ℃ by the extractum of first step gained Radix Tripterygii Wilfordii; Reuse middle polarity organic solvent such as ethyl acetate extraction, wherein the content of extractum is 0.001--2.0g/ml, and extract is washed to neutrality, concentrates to obtain concentrate;
2), Radix Tripterygii Wilfordii or by the extractum of first step gained Radix Tripterygii Wilfordii, carry out the middle polarity organic solvent extraction respectively, acid solution soaks;
With Radix Tripterygii Wilfordii or by the extractum of first step gained Radix Tripterygii Wilfordii, use the middle polarity organic solvent extraction respectively, wherein, the content of extractum in solvent is 0.001--2.0g/ml, after extract concentrates, soak with hydrochloric acid or hydrochloric acid solution respectively, under 0-40 ℃, make Radix Tripterygii Wilfordii lactone alcohol wherein be converted into tripchlorolide.
Described hydrochloric acid, concentration are 0.002-2.0N; Acid solution is selected from sulphuric acid, acetic acid, nitric acid, acetic acid, carbonic acid, phosphoric acid, oxalic acid, Fluohydric acid., citric acid; PH value with hydrochloric acid or hydrochloric acid solution is-2.0-6.0.
Modified technique of the present invention is compared with prior art has following advantage: technological operation of the present invention is easy, and is with low cost, proves through pharmacological evaluation, and the extractum that Radix Tripterygii Wilfordii (herb or part), Radix Tripterygii Wilfordii make is after acid system is handled, and toxicity obviously descends.And present traditional handicraft can't reduce Radix Tripterygii Wilfordii toxicity, and a large amount of clinical practices are restricted.
Description of drawings
Fig. 1: modified technique flow chart of the present invention
The specific embodiment
Enumerate 8 embodiment below, the present invention is further specified, but the present invention is not only limited to these embodiment.The present invention is a material with tripterygium plant-Radix Tripterygii Wilfordii, Tripterygium hypoglaucum and the Tripterygium regelii Spragus et Takeda etc. of China's abundant, and the basic physicochemical property according to its main activity, toxic component by special modified technique, reduces its toxicity at last.
Embodiment 1:
Take by weighing Radix Tripterygii Wilfordii 120g, with the HCl solution soaking of 0.03N, placed 48 hours in 10 ℃ of refrigerators simultaneously, make the Radix Tripterygii Wilfordii lactone alcohol major part in the sample be converted into tripchlorolide, conversion ratio reaches more than 25%.Take out back chloroform extraction 3 times, each 300ml, extracting solution washes with water to neutrality, the reuse anhydrous Na
2SO
4Drying obtains concentrate 0.98g behind the concentrating under reduced pressure.
Embodiment 2:
The first step: take by weighing Radix Tripterygii Wilfordii 650g, add 800ml95% ethanol, reflux, extract, 3 times, each 6 hours, filter, merge ethanol liquid, decompression recycling ethanol gets total composition extractum 12.0g.
Second step: take by weighing Radix Tripterygii Wilfordii 130g, add the 300ml ethyl acetate, reflux, extract, 3 times, each 6 hours, filter, combined ethyl acetate liquid, the reclaim under reduced pressure ethyl acetate must be mainly the extractum 1.56g of middle polarity chemical compound.Extractum 6.0g with the first step obtains adds the 25ml ethyl acetate, supersound extraction 3 times, and each 45min filters, combined ethyl acetate liquid, the reclaim under reduced pressure ethyl acetate gets extractum 1.31g.With 1.56g extractum, after the amount of ethyl acetate dissolving, admix the 4.4g neutral alumina, peroxidating aluminum post, discard ethyl acetate: petroleum ether (1: 9) eluent 250ml, collect ethyl acetate: the eluent 450ml of petroleum ether (1: 1), concentrating under reduced pressure gets sample A.1.31g extractum with after the amount of ethyl acetate dissolving, is admixed the 3.2g neutral alumina, and peroxidating aluminum post discards ethyl acetate: petroleum ether (1: 9) eluent 250ml, collect ethyl acetate: the eluent 450ml of petroleum ether (1: 1), concentrating under reduced pressure gets sample B.
The 3rd step: sample A and sample B are iced the HAc system with the HCl-of 0.4N respectively, handled 36 hours at 4 ℃, fling to solvent under the room temperature, sample A after the extraction and the Radix Tripterygii Wilfordii lactone alcohol among the sample B, major part is converted into tripchlorolide, and conversion ratio all reaches more than 87%.
The 4th step: 1, take by weighing Radix Tripterygii Wilfordii 120g, extractum 4.0g with the HCl-ice HAc solution soaking of 0.4N, placed 36 hours in refrigerator respectively simultaneously, make the Radix Tripterygii Wilfordii lactone alcohol major part in the original sample be converted into tripchlorolide, conversion ratio reaches respectively more than 78% and 80%.Take out back ethyl acetate extraction 3 times, the each 300ml of Radix Tripterygii Wilfordii, the each 20ml of extractum washes to neutrality the reuse anhydrous Na with water behind the merge extractive liquid,
2SO
4Drying obtains concentrate 1.09g and 1.0g respectively behind the concentrating under reduced pressure.
2, take by weighing Radix Tripterygii Wilfordii 130g, add the 300ml ethyl acetate, reflux, extract, 3 times, each 6 hours, filter, combined ethyl acetate liquid, the reclaim under reduced pressure ethyl acetate gets extractum (being mainly the middle polarity chemical compound) 1.67g.Extractum 6.0g with the first step obtains adds the 25ml ethyl acetate, supersound extraction 3 times, and each 45min filters, combined ethyl acetate liquid, the reclaim under reduced pressure ethyl acetate gets extractum 1.10g.With the HCl-ice HAc solution soaking of 0.4N, placed 36 hours in refrigerator simultaneously respectively, make the Radix Tripterygii Wilfordii lactone alcohol major part in the sample be converted into tripchlorolide, conversion ratio all reaches more than 80%.
Embodiment 3:
The first step: Radix Tripterygii Wilfordii 600g, add 700ml95% ethanol, technology gets total composition extractum 12.0g with embodiment one step of 2 mat woven of fine bamboo strips.
Second step: take by weighing Radix Tripterygii Wilfordii 100g, add the 250ml ethyl acetate, technology must be mainly middle polarity compound extract 1.24g with embodiment two steps of 2 mat woven of fine bamboo strips.The extractum 5.6g that the first step is obtained adds the 20ml ethyl acetate, and technology gets extractum 1.16g with embodiment two steps of 2 mat woven of fine bamboo strips.1.24g extractum with after the amount of ethyl acetate dissolving, is admixed the 4.0g neutral alumina, and peroxidating aluminum post discards ethyl acetate: petroleum ether (1: 9) eluent 190ml, collect ethyl acetate: the eluent 400ml of petroleum ether (1: 1), concentrating under reduced pressure gets sample A.1.16g extractum with after the amount of ethyl acetate dissolving, is admixed the 3.0g neutral alumina, and peroxidating aluminum post discards ethyl acetate: petroleum ether (1: 9) eluent 190ml, collect ethyl acetate: the eluent 400ml of petroleum ether (1: 1), concentrating under reduced pressure gets sample B.
The 3rd step: with sample A and sample B, technology is with embodiment 2 mat woven of fine bamboo strips three step, makes sample A after the extraction and the Radix Tripterygii Wilfordii lactone alcohol among the sample B, and major part is converted into tripchlorolide, and conversion ratio all reaches more than 82%.
The 4th step: 1 takes by weighing Radix Tripterygii Wilfordii 120g, extractum 4.0g, and technology makes the Radix Tripterygii Wilfordii lactone alcohol major part in the sample be converted into tripchlorolide with embodiment four steps 1 of 2 mat woven of fine bamboo strips, and conversion ratio reaches respectively more than 77% and 80%.Take out back chloroform extraction 3 times, the each 300ml of Radix Tripterygii Wilfordii, the each 20ml of extractum washes to neutrality the reuse anhydrous Na with water behind the merge extractive liquid,
2SO
4Drying obtains concentrate 0.98g and 0.76g respectively behind the concentrating under reduced pressure.
2 take by weighing Radix Tripterygii Wilfordii 100g, add the 250ml ethyl acetate, and technology gets extractum 1.11g with embodiment four steps 2 of 2 mat woven of fine bamboo strips.The extractum 5.6g that the first step is obtained adds the 20ml ethyl acetate, supersound extraction 3 times, and each 45min filters, combined ethyl acetate liquid, the reclaim under reduced pressure ethyl acetate gets extractum 1.80g.With the HCl-ice HAc solution soaking of 0.4N, placed 36 hours in refrigerator simultaneously respectively, make the Radix Tripterygii Wilfordii lactone alcohol major part in the sample be converted into tripchlorolide, conversion ratio all reaches more than 80%.
Embodiment 4:
The first step: take by weighing Radix Tripterygii Wilfordii 600g, add 700ml95% ethanol, technology is with embodiment one step of 2 mat woven of fine bamboo strips, and decompression recycling ethanol gets extractum 9.0g.
Second step: take by weighing Radix Tripterygii Wilfordii 100g, add the 200ml chloroform, technology must be mainly middle polarity compound extract 1.05g with embodiment two steps of 2 mat woven of fine bamboo strips.The extractum 4.0g that the first step is obtained adds the 20ml chloroform, supersound extraction 3 times, and each 45min filters, combined chloroform liquid, the reclaim under reduced pressure chloroform gets extractum 0.96g.1.05g extractum with after the minimum of chloroform dissolving, is admixed the 3.0g neutral alumina, and peroxidating aluminum post discards ethyl acetate: petroleum ether (1: 9) eluent 180ml, collect ethyl acetate: the eluent 350ml of petroleum ether (1: 1), concentrating under reduced pressure gets sample A.0.96g extractum with after the minimum of chloroform dissolving, is admixed the 3.0g neutral alumina, and peroxidating aluminum post discards ethyl acetate: petroleum ether (1: 9) eluent 180ml, collect ethyl acetate: the eluent 350ml of petroleum ether (1: 1), concentrating under reduced pressure gets sample B.
The 3rd step: with sample A and sample B, technology is with embodiment 2 mat woven of fine bamboo strips three step, makes sample A after the extraction and the Radix Tripterygii Wilfordii lactone alcohol major part among the sample B be converted into tripchlorolide, and conversion ratio reaches more than 88%.
The 4th step: 1, take by weighing Radix Tripterygii Wilfordii 100g, extractum 4.0g, technology makes the Radix Tripterygii Wilfordii lactone alcohol major part of extracting in the sample of back be converted into tripchlorolide with embodiment four steps 1 of 2 mat woven of fine bamboo strips, and conversion ratio reaches respectively more than 70% and 78%.Take out back ethyl acetate extraction 3 times, the each 200ml of Radix Tripterygii Wilfordii, the each 20ml of extractum washes to neutrality the reuse anhydrous Na with water behind the merge extractive liquid,
2SO
4Drying obtains concentrate 1.03g and 0.92g respectively behind the concentrating under reduced pressure.
2, take by weighing Radix Tripterygii Wilfordii 100g, add the 200ml chloroform, technology gets extractum 1.02g with embodiment four steps 2 of 2 mat woven of fine bamboo strips.The extractum 4.0g that the first step is obtained adds the 20ml chloroform, supersound extraction 3 times, and each 45min filters, combined chloroform liquid, the reclaim under reduced pressure chloroform gets extractum 0.90g.With the HCl-ice HAc solution soaking of 0.4N, placed 36 hours in refrigerator simultaneously respectively, make the Radix Tripterygii Wilfordii lactone alcohol major part in the sample be converted into tripchlorolide, conversion ratio reaches more than 80%.
Embodiment 5:
The first step: Radix Tripterygii Wilfordii 600g, add 700ml95% ethanol, technology is with embodiment one step of 2 mat woven of fine bamboo strips, and decompression recycling ethanol gets total composition extractum 13.0g.
Second step: take by weighing Radix Tripterygii Wilfordii 100g, add the 250ml chloroform, technology must be mainly middle polarity compound extract 1.04g with embodiment two steps of 2 mat woven of fine bamboo strips.The extractum 5.8g that the first step is obtained adds the 20ml chloroform, supersound extraction 3 times, and each 45min filters, combined chloroform liquid, the reclaim under reduced pressure chloroform gets extractum 0.95g.With 1.04g extractum with after the minimum of chloroform dissolving, admix the 3.0g neutral alumina, peroxidating aluminum post, discard ethyl acetate: petroleum ether (1: 9) eluent 180ml, technology is with embodiment three steps of 2 mat woven of fine bamboo strips, make sample A after the extraction and the Radix Tripterygii Wilfordii lactone alcohol major part among the sample B be converted into tripchlorolide, conversion ratio reaches more than 82%.
The 4th step: 1, take by weighing Radix Tripterygii Wilfordii 120g, extractum 4.0g, technology makes the Radix Tripterygii Wilfordii lactone alcohol major part in the sample be converted into tripchlorolide with embodiment four steps 1 of 2 mat woven of fine bamboo strips, and conversion ratio reaches respectively more than 74% and 80%.Take out back chloroform extraction 3 times, the each 300ml of Radix Tripterygii Wilfordii, the each 20ml of extractum washes to neutrality the reuse anhydrous Na with water behind the merge extractive liquid,
2SO
4Drying obtains concentrate 0.95g and 0.78g respectively behind the concentrating under reduced pressure.
2, take by weighing Radix Tripterygii Wilfordii 100g, add the 250ml chloroform, technology gets extractum 1.16g with embodiment four steps 2 of 2 mat woven of fine bamboo strips.The extractum 5.8g that second step was obtained adds the 20ml chloroform, supersound extraction 3 times, and each 45min filters, combined chloroform liquid, the reclaim under reduced pressure chloroform gets extractum 0.89g.With the HCl-ice HAc solution soaking of 0.4N, placed 36 hours in refrigerator simultaneously respectively, make the Radix Tripterygii Wilfordii lactone alcohol major part in the sample be converted into tripchlorolide, conversion ratio reaches more than 80%.
Embodiment 6:
The first step: take by weighing Radix Tripterygii Wilfordii 650g, add 800ml95% ethanol, technology is with embodiment one step of 2 mat woven of fine bamboo strips, and decompression recycling ethanol gets general name and divides extractum 10.0g.
Second step: take by weighing Radix Tripterygii Wilfordii 130g, add the 300ml ethyl acetate, technology must be mainly middle polarity compound extract 1.66g with embodiment two steps of 2 mat woven of fine bamboo strips.The extractum 5.0g that the first step is obtained adds the 25ml ethyl acetate, supersound extraction 3 times, and each 45min filters, combined ethyl acetate liquid, the reclaim under reduced pressure ethyl acetate gets extractum 1.01g.1.66g extractum with after the amount of ethyl acetate dissolving, is admixed the 4.0g neutral alumina, and peroxidating aluminum post discards ethyl acetate: petroleum ether (1: 9) eluent 250ml, collect ethyl acetate: the eluent 450ml of petroleum ether (1: 1), concentrating under reduced pressure gets sample A.1.01g extractum with after the amount of ethyl acetate dissolving, is admixed the 3.0g neutral alumina, and peroxidating aluminum post discards ethyl acetate: petroleum ether (1: 9) eluent 250ml, collect ethyl acetate: the eluent 450ml of petroleum ether (1: 1), concentrating under reduced pressure gets sample B.
The 3rd step: with sample A and sample B, technology is with embodiment three steps of 2 mat woven of fine bamboo strips, and sample A after the extraction and the Radix Tripterygii Wilfordii lactone alcohol major part among the sample B are converted into tripchlorolide, and conversion ratio reaches more than 66%.
The 4th step: 1, take by weighing Radix Tripterygii Wilfordii 120g, extractum 5.0g, technology makes the Radix Tripterygii Wilfordii lactone alcohol major part in the original sample be converted into tripchlorolide with embodiment four steps 1 of 2 mat woven of fine bamboo strips, and conversion ratio reaches respectively more than 50% and 57%.。Take out back ethyl acetate extraction 3 times, the each 300ml of Radix Tripterygii Wilfordii, the each 20ml of extractum washes to neutrality the reuse anhydrous Na with water behind the merge extractive liquid,
2SO
4Drying obtains concentrate 1.15g and 1.3g respectively behind the concentrating under reduced pressure.
2, take by weighing Radix Tripterygii Wilfordii 130g, add the 300ml ethyl acetate, technology gets extractum 1.27g with embodiment four steps 2 of 2 mat woven of fine bamboo strips.The extractum 5.0g that the first step is obtained adds the 25ml ethyl acetate, supersound extraction 3 times, and each 45min filters, combined ethyl acetate liquid, the reclaim under reduced pressure ethyl acetate gets extractum 1.03g.With the HCl-ice HAc solution soaking of 0.1N, placed 36 hours in refrigerator simultaneously respectively, make the Radix Tripterygii Wilfordii lactone alcohol major part in the sample be converted into tripchlorolide, conversion ratio reaches more than 60%.
Embodiment 7:
The first step: take by weighing Radix Tripterygii Wilfordii 600g, add 700ml95% ethanol, technology is with embodiment one step of 2 mat woven of fine bamboo strips, and decompression recycling ethanol gets total composition extractum 12.0g.
Second step: take by weighing Radix Tripterygii Wilfordii 100g, add the 250ml ethyl acetate, technology must be mainly middle polarity compound extract 1.20g with embodiment two steps of 2 mat woven of fine bamboo strips.The extractum 6.0g that the first step is obtained adds the 20ml ethyl acetate, supersound extraction 3 times, and each 45min filters, combined ethyl acetate liquid, the reclaim under reduced pressure ethyl acetate gets extractum 1.35g.1.20g extractum with after the amount of ethyl acetate dissolving, is admixed the 3.5g neutral alumina, and peroxidating aluminum post discards ethyl acetate: petroleum ether (1: 9) eluent 190ml, collect ethyl acetate: the eluent 400ml of petroleum ether (1: 1), concentrating under reduced pressure gets sample A.1.35g extractum with after the amount of ethyl acetate dissolving, is admixed the 4.0g neutral alumina, and peroxidating aluminum post discards ethyl acetate: petroleum ether (1: 9) eluent 190ml, collect ethyl acetate: the eluent 400ml of petroleum ether (1: 1), concentrating under reduced pressure gets sample B.
The 3rd step: with sample A and sample B, technology is with embodiment 2 mat woven of fine bamboo strips three step, makes sample A after the extraction and the Radix Tripterygii Wilfordii lactone alcohol major part among the sample B be converted into tripchlorolide, and conversion ratio reaches more than 60%.
The 4th step: 1, take by weighing Radix Tripterygii Wilfordii 120g, extractum 4.0g, technology makes the Radix Tripterygii Wilfordii lactone alcohol major part in the sample be converted into tripchlorolide with embodiment four steps 1 of 2 mat woven of fine bamboo strips, and conversion ratio reaches respectively more than 52% and 59%.Take out back chloroform extraction 3 times, the each 300ml of Radix Tripterygii Wilfordii, the each 20ml of extractum washes to neutrality the reuse anhydrous Na with water behind the merge extractive liquid,
2SO
4Drying obtains concentrate 0.88g and 0.81g respectively behind the concentrating under reduced pressure.
2, take by weighing Radix Tripterygii Wilfordii 100g, add the 250ml ethyl acetate, technology gets extractum 1.24g with embodiment four steps 2 of 2 mat woven of fine bamboo strips.The extractum 6.0g that the first step is obtained adds the 20ml ethyl acetate, supersound extraction 3 times, each 45min filters combined ethyl acetate liquid, the reclaim under reduced pressure ethyl acetate, get extractum 1.63g, with the HCl-ice HAc solution soaking of 0.1N, in refrigerator, placed 36 hours simultaneously respectively, make the Radix Tripterygii Wilfordii lactone alcohol major part in the sample be converted into tripchlorolide, conversion ratio reaches more than 60%.
Embodiment 8:
The first step: Radix Tripterygii Wilfordii 600g, add 700ml95% ethanol, technology is with embodiment one step of 2 mat woven of fine bamboo strips, and decompression recycling ethanol gets total composition extractum 12.0g.
Second step: take by weighing Radix Tripterygii Wilfordii 100g, add the 250ml ethyl acetate, technology must be mainly middle polarity compound extract 1.24g with embodiment two steps of 2 mat woven of fine bamboo strips.The extractum 5.6g that the first step is obtained adds the 20ml ethyl acetate, supersound extraction 3 times, and each 45min filters, combined ethyl acetate liquid, the reclaim under reduced pressure ethyl acetate gets extractum 1.16g.1.24g extractum with after the amount of ethyl acetate dissolving, is admixed the 4.0g neutral alumina, and peroxidating aluminum post discards ethyl acetate: petroleum ether (1: 9) eluent 190ml, collect ethyl acetate: the eluent 400ml of petroleum ether (1: 1), concentrating under reduced pressure gets sample A.1.16g extractum with after the amount of ethyl acetate dissolving, is admixed the 3.0g neutral alumina, and peroxidating aluminum post discards ethyl acetate: petroleum ether (1: 9) eluent 190ml, collect ethyl acetate: the eluent 400ml of petroleum ether (1: 1), concentrating under reduced pressure gets sample B.
The 3rd step: with sample A and sample B, use the HCl solution soaking of 0.03N respectively, placed 48 hours in refrigerator simultaneously, make sample A after the extraction and the Radix Tripterygii Wilfordii lactone alcohol major part among the sample B be converted into tripchlorolide, conversion ratio reaches more than 35%.
The 4th step: 1, take by weighing Radix Tripterygii Wilfordii 120g, extractum 4.0g uses the HCl solution soaking of 0.03N respectively, places 48 hours in refrigerator simultaneously, make the Radix Tripterygii Wilfordii lactone alcohol major part in the sample be converted into tripchlorolide, conversion ratio reaches respectively more than 25% and 30%.Take out back chloroform extraction 3 times, the each 300ml of Radix Tripterygii Wilfordii, the each 20ml of extractum washes to neutrality the reuse anhydrous Na with water behind the merge extractive liquid,
2SO
4Drying obtains concentrate 0.98g and 0.76g respectively behind the concentrating under reduced pressure.
2, take by weighing Radix Tripterygii Wilfordii 100g, add the 250ml ethyl acetate, technology gets extractum 1.11g with embodiment four steps 2 of 2 mat woven of fine bamboo strips.The extractum 5.6g that the first step is obtained adds the 20ml ethyl acetate, supersound extraction 3 times, each 45min filters combined ethyl acetate liquid, the reclaim under reduced pressure ethyl acetate, get extractum 1.80g, use the HCl solution soaking of 0.03N respectively, in refrigerator, placed 48 hours simultaneously, make the Radix Tripterygii Wilfordii lactone alcohol major part in the sample be converted into tripchlorolide, conversion ratio reaches more than 35%.
Under other acid system conditions, processing method is the same.
Overall merit
Show by pharmacological evaluation, compare with extractum with the Radix Tripterygii Wilfordii of non-modified, with the Radix Tripterygii Wilfordii of new technology modification (
Be the tripchlorolide active component)And extractum, its toxicity obviously descends, LD
50By becoming (931.10 ± 0.02) mg/kg, (840.10 ± 0.02) mg/kg by original (279.4 ± 0.02) mg/kg, (252.0 ± 0.02) mg/kg respectively.And this modified technique is easy and simple to handle, and is with low cost, can effectively reduce the toxicity of Radix Tripterygii Wilfordii and extractum, and make full use of the Radix Tripterygii Wilfordii medicine resource, has broad application prospects.
Product of the present invention is used for the treatment of rheumatoid arthritis, chronic nephritis, hepatitis, thrombocytopenic purpura and various dermatoarthritis, the research of aspect such as repels at present after antifertility, antitumor, anti-organ transplantation in the body, has also obtained bigger progress.
Claims (3)
1, the modified technique of Chinese medicine Radix Tripterygii Wilfordii and extract attenuation synergistic thereof is characterized in that
(1), directly by with soaking the Chinese medicine Radix Tripterygii Wilfordii with hydrochloric acid or hydrochloric acid solution respectively, the content of Radix Tripterygii Wilfordii in hydrochloric acid or hydrochloric acid solution is 0.001--1.8g/ml, makes Radix Tripterygii Wilfordii lactone alcohol wherein be converted into tripchlorolide; Or
(2), technology is carried out according to the following steps:
The first step: total composition extracts in the Radix Tripterygii Wilfordii, gets extractum
With polar solvent or middle polarity organic solvent extraction, the amount of Radix Tripterygii Wilfordii in solvent is 0.001--1.8g/ml with Radix et Rhizoma Tripterygii, and sucking filtration reclaims solvent, extractum;
Second step: Radix Tripterygii Wilfordii or by the purification of the extractum of first step gained Radix Tripterygii Wilfordii
With Radix Tripterygii Wilfordii or by the extractum of first step gained Radix Tripterygii Wilfordii, use the middle polarity organic solvent extraction respectively, the amount of extractum is 0.001--1.2g/ml, concentrate, chromatography, separation and purification, diterpene-kind compound;
The 3rd step: the Radix Tripterygii Wilfordii behind the purification or by the acid treatment of the extractum of first step gained Radix Tripterygii Wilfordii
Radix Tripterygii Wilfordii behind the purification or by the extractum of first step gained Radix Tripterygii Wilfordii is got a certain amount of Radix Tripterygii Wilfordii or by the extractum of first step gained Radix Tripterygii Wilfordii, handles with hydrochloric acid or hydrochloric acid solution respectively, and wherein, the content of extractum in acid is 0.001--2.0g/ml;
1), Radix Tripterygii Wilfordii or the 4th step:, carry out acid solution processing, reuse middle polarity organic solvent extraction, washing respectively, must concentrate modifier by the extractum of first step gained Radix Tripterygii Wilfordii;
Get a certain amount of Radix Tripterygii Wilfordii or, soak with hydrochloric acid or hydrochloric acid solution respectively, handle down, make Radix Tripterygii Wilfordii lactone alcohol major part wherein be converted into tripchlorolide at 0-40 ℃ by the extractum of first step gained Radix Tripterygii Wilfordii; Reuse middle polarity organic solvent extraction, wherein the content of extractum is 0.001--2.0g/ml, and extract is washed to neutrality, concentrates to obtain concentrate;
2), Radix Tripterygii Wilfordii or by the extractum of first step gained Radix Tripterygii Wilfordii, use earlier the middle polarity organic solvent extraction, the immersion of reuse acid solution;
With Radix Tripterygii Wilfordii or by the extractum of first step gained Radix Tripterygii Wilfordii, use organic solvent extraction respectively, wherein the content of extractum in solvent is 0.001-2.0g/ml, after extract concentrates, soak with hydrochloric acid or hydrochloric acid solution respectively, under 0-40 ℃, make Radix Tripterygii Wilfordii lactone alcohol wherein be converted into tripchlorolide.
2, modified technique according to claim 1 is characterized in that described Radix Tripterygii Wilfordii is meant Radix Tripterygii Wilfordii herb or part.
3, modified technique according to claim 1 is characterized in that described hydrochloric acid, and concentration is 0.002-2.0N; Acid solution is selected from sulphuric acid, acetic acid, nitric acid, acetic acid, carbonic acid, phosphoric acid, oxalic acid, Fluohydric acid., citric acid; PH value-2.0-6.0 with hydrochloric acid or hydrochloric acid solution.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101361779B (en) * | 2008-09-19 | 2010-09-29 | 马伟光 | Attenuation method of tripterygium wilfordii |
| CN103641884A (en) * | 2013-11-26 | 2014-03-19 | 浙江得恩德制药有限公司 | Method for extracting tripchlorolide from tripterygium wilfordii front segment |
| CN114716503A (en) * | 2022-03-23 | 2022-07-08 | 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 wilfordii chlorine lactonic alcohol |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1141877C (en) * | 2001-09-18 | 2004-03-17 | 武汉绿世纪生物工程有限责任公司 | Application of non-aqueous threewingnut extractive in preparing pesticide |
| CN1299667C (en) * | 2001-09-29 | 2007-02-14 | 华中科技大学 | Nano Chinese medicine threewingnut/Tripterygium prepn and its prepn process |
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2004
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101361779B (en) * | 2008-09-19 | 2010-09-29 | 马伟光 | Attenuation method of tripterygium wilfordii |
| CN103641884A (en) * | 2013-11-26 | 2014-03-19 | 浙江得恩德制药有限公司 | Method for extracting tripchlorolide from tripterygium wilfordii front segment |
| CN103641884B (en) * | 2013-11-26 | 2015-11-18 | 浙江得恩德制药有限公司 | A kind of method extracting Triptolide 12,13-chlorohydrin in trypterygine leading portion medicinal extract |
| CN114716503A (en) * | 2022-03-23 | 2022-07-08 | 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 wilfordii chlorine lactonic alcohol |
| 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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