CN100546590C - Anti-cancer Tengli root extract and its production and use - Google Patents
Anti-cancer Tengli root extract and its production and use Download PDFInfo
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- CN100546590C CN100546590C CNB2005100326685A CN200510032668A CN100546590C CN 100546590 C CN100546590 C CN 100546590C CN B2005100326685 A CNB2005100326685 A CN B2005100326685A CN 200510032668 A CN200510032668 A CN 200510032668A CN 100546590 C CN100546590 C CN 100546590C
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Abstract
The invention discloses a kind of anti-cancer Tengli root extract and its production and use.This anti-cancer Tengli root extract can be by water or organic solvent lixiviate under 70～100 ℃ of temperature, concentrating under reduced pressure, acid adjustment, extraction, polyamide chromatography, solvent elution then, and concentrate drying makes.This anti-cancer Tengli root extract can be used as active component and acceptable drug carrier and makes and be used for the treatment of the oral of various tumors or injection folk prescription and compound preparation.Extracting effect of the present invention is good, antitumor effective ingredient rate of transform height in the crude drug; Adopt the acid adjustment method to remove acid non-soluble substance, improve effect of extracting, the product purity height; Adopt refining the refining of active component that extract of polyamide, effectively purification actinidia root antitumor effective site is convenient to suitability for industrialized production.
Description
Technical field
The present invention relates to field of medicaments, particularly a kind of anti-cancer Tengli root extract and its production and use.
Background technology
Radix Actinidiae Chinensis is the fall leaves root of big liana Fructus actinidiae chinensis (Actinidia chinensis Planch.) of Actinidiaceae, having that detoxifcation is invigorated blood circulation, the effect of clearing away heat-damp and promoting diuresis.Old passing through really waits the people at " Fructus actinidiae chinensis ", Chinese crude drug science and technology, 1981,4 (1): point out in 30 that Radix Actinidiae Chinensis contains ursolic acid, oleanolic acid, vitamin C, saccharide, pigment and aminoacid.People such as Chen Dexuan are at " anti-tumor botanical Radix Actinidiae Chinensis ", Chinese crude drug, and 2004,27 (2): report in 86～87, Radix Actinidiae Chinensis has antitumaous effect, and the intraperitoneal administration of its ethanol extraction is 30～40% to the suppression ratio of S180.People such as Zou friend and mentor are in " Radix Actinidiae Chinensis suppresses the experimentation of tumor cell ", Chinese medicine Leader, Hunan, 1999,5 (4): in 37～38, composition to Radix Actinidiae Chinensis is studied, and finds that its ethyl acetate extract reaches 54% and 60% respectively to leukaemia and colon cancer poorly differentiated adenocarcinoma cell inhibiting rate.Li Hao, Yang Huiping, people such as Yang Fan are in " experimentation that Radix Actinidiae Chinensis suppresses gastric cancer ", the Hebei traditional Chinese medical science, 2004,26 (4): research report in 314～315, the ethyl acetate extract of Radix Actinidiae Chinensis to the inhibitory rate 88.19% of stomach cancer cell, but are done further research to its anti-tumor effective component and method of separating component when 1.3mg/ml concentration.
Publication number is that the Chinese invention patent of CN1478495A, CN1093594A discloses other Chinese medicine of Radix Actinidiae Chinensis compatibility and is used for the treatment of anticancer disease, but its effective ingredient separates and cancer therapeutic applications is not seen relevant patent, also not about the report of actinidia root antitumor effective site and method of separating component, preparation technique.
Summary of the invention
The objective of the invention is to overcome the shortcoming that exists in the prior art, provide a kind of Chinese medicine of purification effectively Radix Actinidiae Chinensis effective site, be convenient to the anti-cancer Tengli root extract preparation method of suitability for industrialized production.
Another object of the present invention is to provide a kind of anti-cancer Tengli root extract by method for preparing.
A further object of the present invention is to provide a kind of purposes of above-mentioned anti-cancer Tengli root extract.
Purpose of the present invention is achieved through the following technical solutions: the preparation method of this anti-cancer Tengli root extract comprises the steps:
(1) water or organic solvent lixiviate Radix Actinidiae Chinensis medical material under 70～100 ℃ of temperature obtains leachate;
(2) leachate being evaporated to ratio of specific heat and weighing 1.03～1.1, also can be that normal pressure concentrates.Then pH value is adjusted to 2～5, leaves standstill, precipitate, obtain supernatant;
(3) use the ethyl acetate extraction supernatant, reclaim solvent, obtain extract;
(4) extract is carried out the polyamide chromatography, the reuse solvent elution obtains eluent;
(5) eluent concentrated, reclaim solvent, drying, obtain anti-cancer Tengli root extract.
In the step (1), described organic solvent is 30～70% (V/V) ethanol; Described Radix Actinidiae Chinensis medical material can be Radix Actinidiae Chinensis block or ground product; Preferred 70～80 ℃ of described extraction temperature.
In the step (1), described lixiviate number of times is 1～3 time; Each extraction time is 1～2 hour; The Radix Actinidiae Chinensis of lixiviate for the first time and the weight ratio of water or organic solvent are 1: 8～10; Be 1: 4～5 with the Radix Actinidiae Chinensis of lixiviate for the third time and the weight ratio of water or organic solvent for the second time.
In the step (2), adopt acid reagent to regulate the pH value of leachate, preferred hydrochloric acid or acetic acid; Described precipitation can adopt filters or the centrifugalize precipitation.
In the step (3), the consumption of described ethyl acetate is 1～3 times of volume of supernatant.
In the step (4), carry out the polyamide chromatography again after can be with extract water-soluble.
In the step (4), carry out the polyamide chromatography again after can be with extract alcohol molten.
In the step (4), extract can be mixed with a small amount of polyamide, carry out the polyamide chromatography again.
In the step (4), can mix, carry out the polyamide chromatography again with the molten back of extract alcohol and with a small amount of polyamide.
In the step (4), the concentration of described solvent is 0～80% (V/V); Preferred 30～70% (V/V) ethanol or acetone; Described solvent elution is ethanol or acetone gradient elution, and fraction collection obtains eluent.
In the step (5), described drying can be vacuum drying, spray drying or lyophilization.
Adopt method for preparing to obtain anti-cancer Tengli root extract; Prepared anti-cancer Tengli root extract effective ingredient (anthraquinone class) content can reach 80～90%.
This anti-cancer Tengli root extract combines with the acceptable drug carrier to make as active component and can be used for treating the oral of various tumors or injection folk prescription and compound preparation.As being the tablet made of adjuvant with starch etc., being the capsule made of adjuvant with the microcrystalline Cellulose, being injection of making of support etc. with mannitol.
The present invention compared with prior art has following advantage and effect:
(1) the present invention to raw medicinal material adopt high temperature, repeatedly, long-time lixiviate, extracting effect is good, antitumor effective ingredient rate of transform height in the raw medicinal material;
(3) adopt the acid adjustment method to remove acid non-soluble substance, improve effect of extracting, the product purity height;
(4) adopt the refining active component that extracts of polyamide, effectively purification actinidia root antitumor effective site is convenient to suitability for industrialized production.
The specific embodiment
Below in conjunction with embodiment the present invention is done further detailed description, but embodiments of the present invention are not limited thereto.
Embodiment 1
With the Radix Actinidiae Chinensis medical material of 1Kg, add the water of 8L, remained under 100 ℃ of temperature lixiviate 60 minutes, filter then and obtain leachate for the first time; The water that adds 4L again in filtering residue remained under 100 ℃ of temperature lixiviate 60 minutes, filtered then and obtained leachate for the second time; Merge leachate twice.
Leachate is concentrated into ratio of specific heat weighs 1.05 (5L), the pH value of leachate is transferred to 2, leave standstill then, the centrifugal precipitation of going, obtain supernatant with 10% hydrochloric acid; With the ethyl acetate extraction supernatant of 5L 2 times, combining extraction liquid reclaims ethyl acetate, obtains extract; Extract is dissolved with the long-pending water gaging of isoploid, and last sample polyamide column carries out chromatography, and reuse 30～70% ethanol carry out gradient elution, the fraction collection eluent; At last with 70% ethanol elution concentrating under reduced pressure, reclaim ethanol, vacuum drying again obtains the anti-cancer Tengli root extract of 28g.The content of its anthraquinone class is 87%.
Embodiment 2
The Radix Actinidiae Chinensis pulverizing medicinal materials of 1Kg is become coarse powder, add the water of 9L, remained under 90 ℃ of temperature lixiviate 90 minutes, filter then and obtain leachate for the first time; The water that adds 5L again in filtering residue remained under 90 ℃ of temperature lixiviate 90 minutes, filtered then and obtained leachate for the second time; Merge leachate twice.
Leachate is concentrated into ratio of specific heat weighs 1.05 (6L), the pH value of leachate is transferred to 3, leave standstill then, the centrifugal precipitation of going, obtain supernatant with 5% hydrochloric acid; With the ethyl acetate extraction supernatant of 12L 2 times, combining extraction liquid reclaims ethyl acetate, obtains extract; Extract with the long-pending amount of isoploid dissolve with ethanol, is mixed sample with 2 times of weight polyamides, and last sample polyamide column carries out chromatography, and reuse 0～50% ethanol carries out gradient elution, the fraction collection eluent; At last 50% ethanol elution normal pressure concentrate is reclaimed ethanol, spray drying again obtains the anti-cancer Tengli root extract of 30g.The content of its anthraquinone class is 83%.
Embodiment 3
The Radix Actinidiae Chinensis pulverizing medicinal materials of 1Kg is become coarse powder, add the water of 10L, remained under 80 ℃ of temperature lixiviate 120 minutes, filter then and obtain leachate for the first time; The water that adds 5L again in filtering residue remained under 80 ℃ of temperature lixiviate 120 minutes, filtered then and obtained leachate for the second time; The water that adds 5L again in filtering residue remained under 80 ℃ of temperature lixiviate 120 minutes, filtered then and obtained leachate for the third time; Merge three times leachate.
Leachate is concentrated into ratio of specific heat weighs 1.03 (10L), the pH value of leachate is transferred to 5, leave standstill then, the centrifugal precipitation of going, obtain supernatant with 20% acetic acid; With the ethyl acetate extraction supernatant of 10L 1 time, combining extraction liquid reclaims ethyl acetate, obtains extract; With extract equal volume amounts water dissolution, last sample polyamide column carries out chromatography, and reuse 30～80% ethanol carry out gradient elution, the fraction collection eluent; At last 80% ethanol elution concentrating under reduced pressure is reclaimed ethanol, lyophilization again obtains the anti-cancer Tengli root extract of 35g.The content of its anthraquinone class is 90%.
Embodiment 4
The Radix Actinidiae Chinensis pulverizing medicinal materials of 1Kg is become coarse powder, add 30% ethanol of 10L, remained under 70 ℃ of temperature lixiviate 120 minutes, filter then and obtain leachate for the first time; 30% ethanol that adds 5L again in filtering residue remained under 70 ℃ of temperature lixiviate 120 minutes, filtered then and obtained leachate for the second time; Merge the secondary leachate.
Leachate is concentrated into ratio of specific heat weighs 1.03 (8L), the pH value of leachate is transferred to 5, leave standstill then, the centrifugal precipitation of going, obtain supernatant with 20% acetic acid; With the ethyl acetate extraction supernatant of 24L 1 time, combining extraction liquid reclaims ethyl acetate, obtains extract; With extract equal volume amounts water dissolution, last sample polyamide column carries out chromatography, and reuse 30～80% acetone carry out gradient elution, the fraction collection eluent; At last 80% acetone eluent normal pressure concentrate is reclaimed acetone, lyophilization again obtains the anti-cancer Tengli root extract of 33g.The content of its anthraquinone class is 92%.
Embodiment 5
With the Radix Actinidiae Chinensis medical material of 1Kg, add 50% ethanol of 9L, remained under 80 ℃ of temperature lixiviate 90 minutes, filter then and obtain leachate for the first time; 50% ethanol that adds 5L again in filtering residue remained under 80 ℃ of temperature lixiviate 90 minutes, filtered then and obtained leachate for the second time; Merge the secondary leachate.
Leachate is concentrated into ratio of specific heat weighs 1.03 (6L), the pH value of leachate is transferred to 3, leave standstill then, the centrifugal precipitation of going, obtain supernatant with 5% hydrochloric acid; With the ethyl acetate extraction supernatant of 12L 2 times, combining extraction liquid reclaims ethyl acetate, obtains extract; With extract equal volume amounts dissolve with ethanol, last sample polyamide column carries out chromatography, and reuse 0～70% ethanol carries out gradient elution, the fraction collection eluent; To wash 70% ethanol at last and take off the liquid concentrating under reduced pressure and reclaim ethanol, spray drying again obtains the anti-cancer Tengli root extract of 30g.The content of its anthraquinone class is 89%.
Embodiment 6
With the Radix Actinidiae Chinensis medical material of 1Kg, add 70% ethanol of 8L, remained under 70 ℃ of temperature lixiviate 60 minutes, filter then and obtain leachate for the first time; 70% ethanol that adds 4L again in filtering residue remained under 70 ℃ of temperature lixiviate 60 minutes, filtered then and obtained leachate for the second time; 70% ethanol that adds 4L again in filtering residue remained under 70 ℃ of temperature lixiviate 60 minutes, filtered then and obtained leachate for the third time; Merge three times leachate.
Leachate is concentrated into ratio of specific heat weighs 1.05 (6L), the pH value of leachate is transferred to 2, leave standstill then, the centrifugal precipitation of going, obtain supernatant with 10% hydrochloric acid; With the ethyl acetate extraction supernatant of 6L 3 times, combining extraction liquid reclaims ethyl acetate, obtains extract; With waiting weight polyamide to mix sample, last sample polyamide column carries out chromatography with extract, and reuse 0～70% acetone carries out gradient elution, the fraction collection eluent; At last 70% acetone eluent normal pressure concentrate is reclaimed acetone, vacuum drying again obtains the anti-cancer Tengli root extract of 36g.The content of its anthraquinone class is 94%.
Embodiment 7
Radix Actinidiae Chinensis effective site suppresses the cell experiment of tumor
Method: with tumor cell (2 * 10
6/ ml), put into 96 aseptic hole culture plates, every hole adds tumor cell suspension 50 μ l, add the RPMI-1640 50 μ l that contain 15% calf serum, the aqueous solution 10 μ l that add the anti-cancer Tengli root extract of embodiment 1～3 preparation, every group 3 hole adds the normal saline vibration mixing of equivalent in the hole, put into 5% CO then
2Incubator (37 ℃) is cultivated 24h, adds tetrazolium salts (MTT) phosphate buffer, and every hole 10 μ l (every ml contains MTT5mg) continue to cultivate 4h, add dimethylsulfone 100ml then, cessation reaction is with enzyme glue immune detection instrument, respectively at wavelength 570nm, the 630nm place measures optical density OD value, result of calculation.
Computing formula:
Result: in gastric cancer RF cell strain and the experiment of rectal cancer LS174T cell strain tumor cell culture, find, anti-cancer Tengli root extract is significantly higher than the water extract (without the water extracting liquid of crossing acid adjustment, extraction and polyamide chromatography) of its medical material to the suppression ratio of two kinds of tumors, along with dosage escalation, tumor killing effect improves.The results are shown in subordinate list 1.
Subordinate list 1 Radix Actinidiae Chinensis effective site is to the inhibition experiment of gastric cancer and rectum cancer cell
Embodiment 8
Radix Actinidiae Chinensis effective site suppresses the zoopery of tumor
Method: get mice S180 tumor piece, dilution and adjustment cell number (1 * 10 after the homogenate
7/ ml), it is subcutaneous to inject the right axil of mice with 0.2ml, and random packet administration after 24 hours wherein, is organized gastric infusion in contrast with normal saline; CTX (cyclophosphamide) organizes intraperitoneal administration; The anti-cancer Tengli root extract of water extract of Radix Actinidiae Chinensis medical material (without the water extracting liquid of crossing acid adjustment, extraction and polyamide chromatography) and embodiment 3,6 preparations is gastric infusion, the execution mice of weighing in the 11st day is got the tumor piece, claim that tumor is heavy and survey the tumor volume, calculate tumour inhibiting rate.
The result: the water extract of Radix Actinidiae Chinensis medical material is 33.6% to the S180 tumour inhibiting rate, and the tumour inhibiting rate of the anti-cancer Tengli root extract of isodose the present invention's preparation is 64.6%, there were significant differences with matched group, and not remarkable with the difference of CTX, points out it to have good antitumor action.The results are shown in subordinate list 2.
Subordinate list 2 Radix Actinidiae Chinensis effective sites are to the inhibition zoopery of S180 sarcoma
Claims (9)
1, a kind of preparation method of anti-cancer Tengli root extract is characterized in that comprising the steps:
(1) water or volume ratio are ethanol lixiviate Radix Actinidiae Chinensis medical material under 70～100 ℃ of temperature of 30～70%, obtain leachate;
(2) leachate being evaporated to ratio of specific heat and weighing 1.03～1.1, also can be that normal pressure concentrates; Then pH value is adjusted to 2～5, leaves standstill, precipitate, obtain supernatant;
(3) use the ethyl acetate extraction supernatant, reclaim solvent, obtain extract;
(4) extract being carried out the polyamide chromatography, is 0～80% ethanol or acetone gradient elution with volume ratio, the eluent that fraction collection obtains;
(5) eluent concentrated, reclaim solvent, drying, obtain anti-cancer Tengli root extract.
2, the preparation method of anti-cancer Tengli root extract according to claim 1 is characterized in that: in the step (1), described extraction temperature is 70～80 ℃.
3, the preparation method of anti-cancer Tengli root extract according to claim 1, it is characterized in that: in the step (1), the lixiviate number of times is 3 times, each extraction time is 1～2 hour, the Radix Actinidiae Chinensis of lixiviate for the first time and water or volume ratio are that 30～70% alcoholic acid weight ratio is 1: (8～10), for the second time and the Radix Actinidiae Chinensis of lixiviate for the third time and water or volume ratio be that 30～70% alcoholic acid weight ratio is 1: (4～5).
4, the preparation method of anti-cancer Tengli root extract according to claim 1 is characterized in that: in the step (2), adopt hydrochloric acid or acetic acid to regulate the pH value of leachate.
5, the preparation method of anti-cancer Tengli root extract according to claim 1 is characterized in that: in the step (3), the consumption of described ethyl acetate is 1～3 times of volume of supernatant.
6, the preparation method of anti-cancer Tengli root extract according to claim 1, it is characterized in that: in the step (4), earlier with extract water-soluble or pure molten mix with a small amount of polyamide or pure molten after mix with a small amount of polyamide again, and then carry out the polyamide chromatography.
7, the preparation method of anti-cancer Tengli root extract according to claim 1 is characterized in that: in the step (4), the employing volume ratio is 30～70% ethanol or acetone gradient elution, and fraction collection obtains eluent.
8, a kind of anti-cancer Tengli root extract according to each described method preparation of claim 1～7.
9, the application of anti-cancer Tengli root extract according to claim 8 is characterized in that: anti-cancer Tengli root extract is combined with the acceptable drug carrier to make as active component be used for the treatment of the oral of various tumors or injection folk prescription and compound preparation.
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| CN100546590C true CN100546590C (en) | 2009-10-07 |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN100506233C (en) * | 2005-12-09 | 2009-07-01 | 中国科学院化学研究所 | Radix actinidiae argutae extract and its anticanceruse |
| CN101518556B (en) * | 2009-04-07 | 2012-06-20 | 浙江大学 | Polysaccharide with antitumor and immunologic adjuvant function as well as preparation method and purpose thereof |
| CN102526140A (en) * | 2012-02-28 | 2012-07-04 | 陕西中医学院 | Chinese actinidia root extract and preparation method thereof |
| CN104825507B (en) * | 2015-05-07 | 2018-03-20 | 中国医科大学附属盛京医院 | A kind of extract of yangtao actinidia root and its application in treatment bile duct cancer drug is prepared |
| CN113583080B (en) * | 2021-08-13 | 2022-09-30 | 中南大学 | Compound and preparation method and application thereof |
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2005
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Non-Patent Citations (2)
| Title |
|---|
| 藤梨根对胃癌细胞抑制作用的实验研究. 李昊，杨慧萍，杨凡，鲁小青，张虹，陈百先.河北中医，第26卷第（4）期. 2004 |
| 藤梨根对胃癌细胞抑制作用的实验研究. 李昊，杨慧萍，杨凡，鲁小青，张虹，陈百先.河北中医，第26卷第（4）期. 2004 * |
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