CN101028322B - Use of Maoliefengdou extract for preparing anti-cancer medicine - Google Patents
Use of Maoliefengdou extract for preparing anti-cancer medicine Download PDFInfo
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- CN101028322B CN101028322B CN2007100389611A CN200710038961A CN101028322B CN 101028322 B CN101028322 B CN 101028322B CN 2007100389611 A CN2007100389611 A CN 2007100389611A CN 200710038961 A CN200710038961 A CN 200710038961A CN 101028322 B CN101028322 B CN 101028322B
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- cancer
- rhizoma petasitis
- petasitis tricholobi
- carcinoma
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Abstract
An application of the Japanese butterbur's extract (bakkenolide D, B, IVa and IIIa) in preparing the medicines for preventing and treating liver cancer, lung cancer, cervical cancer, intestinal cancer, kidney cancer, bread cancer, etc is disclosed.
Description
Technical field
The present invention relates to medical technical field, be used to prepare the purposes of treatment and prophylaxis of tumours disease medicament in particular to a kind of Rhizoma Petasitis tricholobi rhizome extract and preparation method thereof and this extract
Background technology
Rhizoma Petasitis tricholobi Petasites tricholobus Franch. belongs to the butterbur plant, and its rhizome is at the detoxification and removing stasis that is used among the people, and traumatic injury, fracture snakebite etc. are controlled in external application.For developing this plant inventor this plant has been carried out chemical constituent and pharmacological research than system, once four Rhizoma Petasitis tricholobi lactone noval chemical compounds in the Rhizoma Petasitis tricholobi had been reported, and the Rhizoma Petasitis tricholobi lactone compounds be used for the treatment of may with the purposes (patent publication No.: CN1844114) of histamine and/or leukotriene-related disease, but up to the present, do not see that as yet Rhizoma Petasitis tricholobi and extract thereof have the report of anti-tumor activity.
Summary of the invention
The purpose of this invention is to provide a kind of Petasites tricholobus franch extract that extraction obtains from Rhizoma Petasitis tricholobi and preparation method thereof, through in vitro tests, the Petasites tricholobus franch extract that makes, hepatocarcinoma, pulmonary carcinoma, cervical cancer, intestinal cancer, renal carcinoma, oral cavity epidermoid carcinoma, breast cancer tumour cell strain there is the obvious suppression effect, therefore, Petasites japonicus extract of the present invention can be used for treating tumor disease.These tumor diseases preferably include and refer to hepatocarcinoma, pulmonary carcinoma, cervical cancer, intestinal cancer, renal carcinoma, oral cavity epidermoid carcinoma, breast carcinoma.The prepared Petasites tricholobus franch extract of the present invention has the purposes that can be used for preparing the anti-tumor disease medicine.
The present invention finishes by following technical solution, and particular content comprises: the preparation of Petasites tricholobus franch extract and antitumor pharmacology activity experiment.
(1) preparation of Petasites tricholobus franch extract
The preparation method of Petasites tricholobus franch extract of the present invention is made up of following steps successively:
(1) extracts
After the Rhizoma Petasitis tricholobi rhizome is pulverized, with the aquiferous ethanol of 20-95% extract extracting solution, precipitation is abandoned in centrifugal or filtration behind the extracting solution concentrating under reduced pressure, supernatant or filtrate are standby;
The method that aquiferous ethanol extracts is that heat is carried, merceration or percolation; When adopting said method to extract alcoholic acid volumetric usage be the crude drug quality 5-15 doubly;
(2) absorb-elute
After impurity was removed in the moisture lower alcohol drip washing that above-mentioned supernatant or filtrate are carried out the absorption of polystyrene type porous adsorbent resin, water-50%, the 4-12 that uses sample crude drug quality doubly measured the aqueous C of the 60-95% of volume
1～C
5The lower alcohol eluting;
(3) concentrated, dry
The eluent concentrating under reduced pressure, drying is Petasites tricholobus franch extract of the present invention.
The dry rhizome of Rhizoma Petasitis tricholobi is carried or merceration with 20～95% ethanol heat, extracting solution concentrates the centrifugal or filtration in back, get supernatant or filtrate and carry out macroporous resin adsorption, impurity is removed in the moisture lower alcohol drip washing of elder generation's water-50%, the moisture lower alcohol eluting of reuse 60-95%, eluent is concentrated to there not being alcohol, and drying is Petasites tricholobus franch extract of the present invention; Or with concentrated solution by the polyamide chromatograph, water, 50% ethanol elution are collected 50% ethanol elution successively, concentrate, drying also can get Petasites tricholobus franch extract.
The Petasites tricholobus franch extract component analysis that obtains: through silica gel column chromatography repeatedly, obtains 4 and measure big chemical compounds, these 4 structural general formula are as follows:
Be Rhizoma Petasitis tricholobi lactone D, Rhizoma Petasitis tricholobi lactone B, Rhizoma Petasitis tricholobi lactone IVa and Rhizoma Petasitis tricholobi lactone IIIa.
Wherein:
Chemical compound title R
1R
2
Bakkenolide D. OCOC (CH
3)=CHCH
3OCOCH
3
Bakkenolide B. OCOC (CH
3)=CHCH
3OCOCH
3
Rhizoma Petasitis tricholobi lactone IVa OCOCH=CHSCH
3OCOCH=C (CH
3)
2
Rhizoma Petasitis tricholobi lactone IIIa OCOCH=CHSCH
3OCOC (CH
3)=CHCH
3
So, Petasites tricholobus franch extract mainly is Rhizoma Petasitis tricholobi lactone D, Rhizoma Petasitis tricholobi lactone B, the mixture of Rhizoma Petasitis tricholobi lactone IVa and Rhizoma Petasitis tricholobi lactone IIIa, through the HPLC quantitative assay, the content summation of these 4 chemical compounds accounts for more than 50% of total extract, and Rhizoma Petasitis tricholobi lactone D, Rhizoma Petasitis tricholobi lactone B, the content ratio of Rhizoma Petasitis tricholobi lactone IVa and Rhizoma Petasitis tricholobi lactone IIIa is: 0.1-5: 0.1-5: 1-9: 1-9.
(2) biological activity test: be subjected to the reagent thing that the inhibitory action of tumor cell in vitro growth is subjected to the reagent thing: Petasites tricholobus franch extract (being numbered 1), Rhizoma Petasitis tricholobi lactone D (being numbered 2), Rhizoma Petasitis tricholobi lactone B (being numbered 3), Rhizoma Petasitis tricholobi lactone IVa (being numbered 4), Rhizoma Petasitis tricholobi lactone IIIa (being numbered 5) medicine preparation: will be subjected to the reagent thing to be dissolved in dimethyl sulfoxide (DMSO) and be made into storing solution, and dilute 100 times with the RPMI1640 cell culture medium that contains 10% hyclone during use.The positive control medicine is selected 5-fluorouracil (5-FU), and compound method is with being subjected to the reagent thing.
Tumor cell line: human lung cancer cell A549, human cervical carcinoma cell Hela, human large intestine cancer cell HCT116, human liver cancer cell HepG2, human liver cancer cell BEL-7402, human renal carcinoma cell RXF-631L, human mouth epidermoid carcinoma cell KB, people's mammary gland adenocarcinoma cell MCF-7.
Assay method: will grow into each cell strain trypsinization of logarithmic (log) phase, and make cell suspension with the RPMI1640 culture medium that contains 10% hyclone, with every hole 10
-4The concentration of individual cell is inoculated on the 96 porocyte culture plates, every hole 200 μ l.Behind the 24h, remove culture medium, add and contain the culture medium that is subjected to the reagent thing, the final concentration that is subjected to the reagent thing is 1 * 10
-3-10 μ g/ml, the final concentration of 5-FU are 0.01-10 μ g/ml.Do not add in the matched group and be subjected to the reagent thing.Continue to cultivate after 3 days, stop cultivating, detect cell survival rate with mtt assay.It is 0.5mg/ml that preceding every hole adding MTT solution (5mg/ml) the 10 μ l of termination cultivation make the MTT final concentration, at 37 ℃, and 5%CO
2Incubator continue to hatch 4h after, stop to cultivate, inhale and remove supernatant.Every hole adds 100 μ l lytic agents (10%SDS, 5% isopropyl alcohol, 0.01M HCl), in 37 ℃ of fully dissolvings down, after microscopically is observed the whole dissolvings of crystallization, measures each hole absorbance (OD) value in the 550nm place on microplate reader, calculates cell survival rate.Cell survival rate (%)=be subjected to reagent thing OD value/matched group OD value * 100%., calculate and be subjected to the half-inhibition concentration (IC of reagent thing the mapping of dosage logarithm with cell survival rate each tumor cell
50).Result: see Table 1
Table 1 is subjected to the reagent object to suppress the effect of growth of tumour cell outward
Result's demonstration is subjected to the reagent thing to being tried tumor cell the obvious suppression effect to be arranged all, and particularly for 2 kinds of hepatoma carcinoma cell, inhibitory action is very obvious, and effect is better than 5-FU.The result also shows, Petasites tricholobus franch extract, Rhizoma Petasitis tricholobi lactone IVa, and the effect of Rhizoma Petasitis tricholobi lactone IIIa is better than Rhizoma Petasitis tricholobi lactone D and Rhizoma Petasitis tricholobi lactone B.
The specific embodiment
Providing embodiment below further specifies the present invention but is not construed as limiting the invention.
Embodiment 1
Rhizoma Petasitis tricholobi dry rhizome 20-95% ethanol heating extraction, first the solvent consumption be about the crude drug amount 5-10 doubly, or with Rhizoma Petasitis tricholobi rhizome 20-95% ethanol percolate extraction, the solvent consumption make a living dose 5-15 doubly, merge percolate, and concentrating under reduced pressure gets concentrated solution to certain volume, the centrifuged supernatant or the filtrate of concentrated solution pass through macroporous resin adsorption, (macroporous resin is for being the polystyrene type porous adsorbent resin of bridging materials with styrene, as: D101, D201, ZTC-1, AB-8, D4020,860021, HP20, SP825 type macroporous resins etc.), impurity is removed in the aquiferous ethanol drip washing of water-50%; Reuse concentration is the moisture lower alcohol eluting of 60-95%, and lower alcohol is C1～C5 alcohols such as methanol, ethanol or propanol, its consumption make a living dose 4-12 doubly, collect the lower alcohol eluent, being concentrated into does not have alcohol, drying is Petasites tricholobus franch extract of the present invention.
Embodiment 2
50 kilograms of Rhizoma Petasitis tricholobi dry rhizomes drop into the percolation jar, add 250 liter of 80% ethanol, soak 12 hours, percolation merges percolate, and concentrating under reduced pressure to 30 liter, centrifugal, cross macroporous adsorbent resin SP825 (the green BAICAO in Beijing company) then, remove impurity with 300 liter of 40% ethanol drip washing; 300 liter of 70% ethanol elution of reuse collected eluent, and the concentrating under reduced pressure after drying gets 0.25 kilogram of Petasites tricholobus franch extract of the present invention.
The main active of Petasites tricholobus franch extract of the present invention is Rhizoma Petasitis tricholobi lactone D (bakkenolide-D), Rhizoma Petasitis tricholobi lactone B (bakkenolide-B), Bakkenolide I. Va (bakkenolide-IIIa), Rhizoma Petasitis tricholobi lactone (bakkenolide-IIIa), four total content is up to more than 50%, through in vitro tests, can effectively prevent curing oncoma, particularly hepatocarcinoma, pulmonary carcinoma, cervical cancer, intestinal cancer, renal carcinoma, oral cavity epidermoid carcinoma, breast carcinoma can be used for preparing the purposes of preventing and treating tumour medicine so Petasites tricholobus franch extract has.
Claims (1)
1. the application of Petasites tricholobus franch extract in the anti-hepatocarcinoma of preparation, pulmonary carcinoma, cervical cancer, intestinal cancer, renal carcinoma, oral cavity epidermoid carcinoma, breast cancer medicines, the preparation method of said Petasites tricholobus franch extract is as follows:
(1) extracts
After the Rhizoma Petasitis tricholobi rhizome is pulverized, with the 20-95% aquiferous ethanol extract extracting solution, precipitation is abandoned in centrifugal or filtration behind the extracting solution concentrating under reduced pressure, supernatant or filtrate are standby;
(2) absorb-elute
After impurity was removed in the moisture lower alcohol drip washing that above-mentioned supernatant or filtrate are carried out the absorption of polystyrene type porous adsorbent resin, water-50%, the 4-12 that uses sample crude drug quality doubly measured the aqueous C of the 60-95% of volume
1～C
5The lower alcohol eluting;
(3) concentrated, dry
With the eluent concentrating under reduced pressure, drying promptly gets Petasites tricholobus franch extract.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100389611A CN101028322B (en) | 2007-04-02 | 2007-04-02 | Use of Maoliefengdou extract for preparing anti-cancer medicine |
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|---|---|---|---|
| CN2007100389611A CN101028322B (en) | 2007-04-02 | 2007-04-02 | Use of Maoliefengdou extract for preparing anti-cancer medicine |
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| CN101028322A CN101028322A (en) | 2007-09-05 |
| CN101028322B true CN101028322B (en) | 2011-08-03 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020141083A1 (en) * | 2018-12-31 | 2020-07-09 | Max Zeller Soehne Ag | Combination of cimicifuga and petasites extracts for anticancer therapy and prophylaxis |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102648927A (en) * | 2011-02-25 | 2012-08-29 | 苏州宝泽堂医药科技有限公司 | Production technology of petasites lactone compound |
| CN102649783A (en) * | 2011-02-25 | 2012-08-29 | 苏州宝泽堂医药科技有限公司 | Bakkenolide compounds and extraction process thereof |
| CN102846484B (en) * | 2012-04-20 | 2014-04-16 | 朱丹 | Application of Petasites tatewakianus Kitam lactone in preparing anti-ultraviolet compositions |
| CN110585255A (en) * | 2016-12-12 | 2019-12-20 | 中国人民解放军第二军医大学 | Application of petasites tricholobus franch extract in preparation of medicines or health-care food for preventing and treating anoxia diseases |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003088985A3 (en) * | 2002-04-22 | 2004-02-26 | Weber & Weber Gmbh & Co Kg | Use of compositions containing petasites for treating diseases |
| CN1844114A (en) * | 2006-04-27 | 2006-10-11 | 中国人民解放军第二军医大学 | Fukinanolide compounds and pharmaceutical application thereof |
-
2007
- 2007-04-02 CN CN2007100389611A patent/CN101028322B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003088985A3 (en) * | 2002-04-22 | 2004-02-26 | Weber & Weber Gmbh & Co Kg | Use of compositions containing petasites for treating diseases |
| CN1844114A (en) * | 2006-04-27 | 2006-10-11 | 中国人民解放军第二军医大学 | Fukinanolide compounds and pharmaceutical application thereof |
Non-Patent Citations (2)
| Title |
|---|
| Tian-Shung Wu et al.The bakkenolides from the root of Petasites formosanus and their cytotoxicity.《Chem.Pharm.Bull.》.1999,第47卷(第3期),第375-382页. * |
| 王玉亮等.毛裂蜂斗菜根茎的化学成分及抗炎活性.《第二军医大学学报》.2006,第27卷(第11期),第1210-1213页. * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020141083A1 (en) * | 2018-12-31 | 2020-07-09 | Max Zeller Soehne Ag | Combination of cimicifuga and petasites extracts for anticancer therapy and prophylaxis |
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