CN105267619B - The anti-tumor Chinese medicine compound extract that a kind of suppression ras proto-oncogenes are overexpressed - Google Patents
The anti-tumor Chinese medicine compound extract that a kind of suppression ras proto-oncogenes are overexpressed Download PDFInfo
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Abstract
The present invention relates to a kind of Chinese medicinal compound extract, and its preparation method and application, which is made of ginseng, ophiopogon.Some researches show that about 90% cancer of pancreas, 50% colon cancer, 30% 40% adenocarcinomas of lung and 5% 40% leukaemia are as caused by the overexpression of ras genes, ras genes become the target for the screening cancer therapy drug generally acknowledged.A kind of preparation process of Chinese medicinal compound extract disclosed by the invention, compared with its decocting treatment fluid, the Chinese medicinal compound extract toxicity which is prepared is low, and the effect for suppressing ras gene pathway excessive activations is more preferable.In addition, present invention also offers a kind of new application of Chinese medicinal compound extract, it can apply in suppressing ras proto-oncogenes and being overexpressed and be applied in the antitumor drug for suppressing to be overexpressed after ras Oncogene Mutations is prepared.
Description
Technical field
The present invention relates to a kind of Chinese medicinal compound extract and its preparation process, and concretely the Chinese medicine compound prescription is by ginseng, wheat
Winter forms, and in particular to application of the Chinese medicinal compound extract in suppressing ras proto-oncogenes and being overexpressed and to suppress ras former preparing
Application in the antitumor drug being overexpressed after oncogene mutation, belongs to field of antineoplastic medicaments.
Background technology
At present, malignant tumour has become the number one killer of the mankind, the serious health for endangering the mankind, world health group
The data knitted shows that the morbidity of annual whole world tumour is 10,000,000 people, dead about 7,000,000 people, and the annual cancer in China is annual
New cases are 2,000,000, and what it is because of cancer mortality is about 1,400,000, in 5 people of China's death, that is, has 1 people to die of cancer.
About 30% human tumor is due to that activation causes RAS protein expression levels to increase to make after ras Oncogene Mutations
Into.It if RAS albumen is continuously in the state of activation, can be combined with the effect protein in downstream, pass the signal along to downstream signal member
Part, causes the abnormality proliferation of cell, leads oncogenic generation, and the antineoplaston using ras signal paths as target has become swollen
The therapeutic research hotspot of knurl, therefore, ras proto-oncogenes become the target for the screening decorrelation malignant tumor medicine generally acknowledged.
RAS albumen has important work in terms of several signal of interest paths for participating in adjusting normal cell proliferation are regulated and controled
With so using the signal path that RAS albumen and RAS albumen are regulated and controled as the oncotherapy of target, in treatment by ras proto-oncogenes
There is very important value in tumour caused by mutation.The advanced renal cell carcinoma of U.S. FDA approval on December 20th, 2005
Curative Nexavar, its direct target is exactly Ras signal pathways, can inhibit Raf/MEK/ERK signal transduction pathway, is directly pressed down
Tumour growth processed；Kinase inhibitor such as ID1839 and OSI774 using ras effects sub-channel as target, it can effectively be blocked
The activity of EGFR tyrosine kinase simultaneously can also suppress other members of ERBB protein families to a certain degree, but these are not straight
Connect the medicine for ras.Farnesylation inhibitors using ras albumen as target can also suppress other most of albumen
Farnesylation (some normal human's albumen may be included), but there is also the problem of poor specificity, therefore, exploitation is safe and efficient
The antitumor drugs for the treatment of ras proto-oncogene excessive activations be the task of top priority.
China's Chinese medicine achieves certain curative effect in terms of clinical cancer therapy, and toxicity is relatively low, for tumour prevention and control
Treatment opens a Tiao Xin roads.
Ginseng be Araliaceae ginseng (Panax ginseng C.A.Mey) drying root and rhizome, chief active into
It is divided into ginsenoside and panaxan.Ginseng have reinforce vital energy, reinforce the spleen to benefit the lung, promoting production of body fluid and inducing sedation of the mind intelligence development, it is antifatigue, it is antitumor,
Hemorrhagic shock, reducing blood lipid and other effects.
Ophiopogon is Liliaceae herbaceos perennial ophiopogon (Ophiopogon japonicus (Thumb) Ker-Gawl)
Dried root.Ophiopogon cold nature, sweet in flavor, slight bitter, has, and has enriching yin strengthening the essence, yin-nourishing QI invigorating, relieving restlessness that clears away heart-fire, relaxes bowel
Effect, is one of conventional Chinese medicine.Ophiopogon has extensive pharmacological action, such as resists myocardial ischemia, anti-arrhythmia, anti-aging,
It is antitumor, the effect such as immunological regulation.
So far, no document report is crossed can suppress ras Oncogene Mutations containing ginseng, the Chinese medicine compound prescription of ophiopogon
Overexpression afterwards, in the market also not with containing ginseng, the Chinese medicinal compound extract for raw material of ophiopogon can it is obligate suppress by
Antitumor drug caused by being overexpressed after ras Oncogene Mutations.And the present invention is arduous according to ginseng, the characteristic of ophiopogon, process
Effort, by largely screening analysis, have developed a kind of Chinese medicine compound prescription for the tumour for treating ras proto-oncogene excessive activations, in this
Medicine compound achievees the purpose that to treat tumour by lowering the excessive activations of proto-oncogene Ras paths, and toxicity is low, and no teratogenesis is made
With suitable for the application in the related anti-malignant tumor medicine as caused by being overexpressed after ras Oncogene Mutations is prepared, Ke Yiyu
Pharmaceutically acceptable auxiliary material is mixed and made into various formulations to any type, is expected to exploitation into the antitumor drug of a new generation.
Found during the Chinese medicinal compound extract is prepared, only decocting processing can significantly inhibit ras genes and lead to
The excessive activation on road, but toxicity is big, and security is poor.Therefore, the disclosure of the invention preparation work of a kind of Chinese medicinal compound extract
Skill, refluxing extraction is carried out to the compound with absolute ethyl alcohol, and on the basis of alcohol extracting, by macroreticular resin to it into advancing one
Step isolates and purifies, and the component after isolating and purifying not only remains water decoction and suppresses the effect that ras proto-oncogenes are overexpressed, and greatly
The big toxicity for reducing Chinese medicine compound prescription water decoction, therefore effect is more preferable, can prepare the anti-phase of suppression ras proto-oncogene overexpressions
Close and applied in the medicine of malignant tumour.
The content of the invention
The object of the present invention is to provide a kind of preparation process of Chinese medicinal compound extract, concretely the Chinese medicine compound prescription is by people
Ginseng, ophiopogon composition, high to provide a kind of antitumor activity, small toxicity, cheap Chinese medicinal compound extract, of the invention is another
Purpose is to provide a kind of new application of Chinese medicinal compound extract, and in particular to the Chinese medicinal compound extract is suppressing ras original cancer bases
Application because of the application in overexpression and in the antitumor drug for suppressing to be overexpressed after ras Oncogene Mutations is prepared.
The present invention is practiced by following technical proposals.
A kind of Chinese medicinal compound extract of the present invention, the wherein Chinese medicine compound prescription are made of ginseng, ophiopogon, which answers
Formula extraction is prepared by following method：Ginseng, ophiopogon are taken, adds ethanol solution, immersion, is heated to reflux, and filters, and decompression is dense
Contracting, obtains total medicinal extract；Total medicinal extract is completely dissolved with distilled water, upper macroporous resin column, adsorbs 24h, afterwards with elution, wash
De- liquid is concentrated into thick paste, dry, to obtain the final product.
A kind of Chinese medicinal compound extract of the present invention, is made using ginseng, ophiopogon of raw material, the quality of the bulk pharmaceutical chemicals
Than for 4.5~9：4.5~9.
A kind of Chinese medicinal compound extract of the present invention, is made using ginseng, ophiopogon of raw material, the quality of the bulk pharmaceutical chemicals
Than for 9：4.5.
A kind of Chinese medicinal compound extract of the present invention, the ethanol solution during refluxing extraction is absolute ethyl alcohol.
A kind of Chinese medicinal compound extract of the present invention, the macroreticular resin are D101 type macroreticular resins.
A kind of Chinese medicinal compound extract of the present invention, washing when the total medicinal extract of the Chinese medicine compound prescription isolates and purifies elution
De- amount is 10 times of column volumes.
A kind of Chinese medicinal compound extract of the present invention, the total medicinal extract of Chinese medical extract are used when isolating and purifying
Eluent is ethanol solution.
A kind of Chinese medicinal compound extract of the present invention, when the total medicinal extract of Chinese medical extract is eluted when isolating and purifying
Ethanol solution be absolute ethyl alcohol.At this time, while Chinese medicinal compound extract drug effect is ensured, toxicity is hardly shown.
The present invention is Chinese medicinal compound extract, and the tumour is as caused by being overexpressed after ras Oncogene Mutations.
Application of the present invention for Chinese medicinal compound extract in suppressing ras proto-oncogenes and being overexpressed.
Chinese medicinal compound extract of the present invention is preparing the application in treating antitumor drug, can be by Chinese medicine compound prescription
Extract addition pharmaceutically acceptable carrier such as filler, lubricant, wetting agent, promotion absorbent, absorption carrier, dilution
Agent, excipient etc., can add sweetener, flavouring agent etc. if necessary, be prepared into injection, powder, granule, pulvis, pill, mouth
Take a variety of formulations of the medicines such as liquid, tablet, this be it will be appreciated by those skilled in the art that.
Beneficial effect：The present invention provide it is a kind of treat ras proto-oncogenes overexpression tumour Chinese medicinal compound extract and
It is preparing the application in treating associated malignancies medicine, and the advantages of following has been compared with existing technology：
(1) the Chinese medicinal compound extract effect prepared using the preparation process in the present invention is good, can significantly reduce toxicity, make
The death rate that assessment suppresses the model Caenorhabditis elegans of ras proto-oncogene path excessive activation medicines reduces, WT ratios
Rise.
(2) Chinese medicinal compound extract prepared using the preparation process in the present invention can significantly inhibit ras proto-oncogenes
It is overexpressed, can be applied in treatment tumour medicine as caused by being overexpressed ras proto-oncogenes is prepared.
(3) the prescription proportioning in Chinese medicine compound prescription between ginseng, ophiopogon can rationally be changed according to Chinese prescription principle, be utilized
It still can significantly lower ras proto-oncogene excessive activations to Chinese medicinal compound extract prepared by the preparation process in the present invention,
With antitumor activity.
(4) Chinese medicinal compound extract prepared using the preparation process in the present invention only acts on the mutant of overexpression
RAS albumen, and on wild type RAS albumen without influence.Chinese medicinal compound extract is prompted to apply upper safety, it is thin that it acts only on cancer
Born of the same parents, without acting on normal cell.From now on, we are expected to apply Chinese medicinal compound extract in decorrelation malignant tumor medicine
Preparation in, wherein, the tumour is as caused by the overexpression of ras proto-oncogenes, so as to be provided more for such tumor patient
Good treatment, the treatment for some difficult cancers provide new approach.
(5) Chinese medicinal compound extract significantly lowers ras proto-oncogenes overexpression, is not caused by mutagenesis, not teratogenesis.
(6) due to ras signal paths from nematode to people it is highly conserved, in Caenorhabditis elegans let-60 gene codes protect
The RAS albumen kept, it has 83% homology compared with the RAS albumen of the mankind.If ras is mutated, in Caenorhabditis elegans
In nematode can be caused to produce more vaginal orifices, be then the malignant proliferation for causing cell in the mankind, lead oncogenic generation.Therefore, mould
Formula biology Caenorhabditis elegans (Caenorhabditis elegans) Ras/MAPK signal path excessive activation type mutant quilts
It is widely used in the antitumor drug of screening suppression ras proto-oncogene path excessive activations as test population.Therefore, it is of the invention
Employ Caenorhabditis elegans mutant strain and suppress the model of ras proto-oncogene path excessive activation medicines as assessment, and draw
Chinese medicinal compound extract, which is overexpressed ras proto-oncogenes, obvious downward effect, has that to suppress ras proto-oncogene paths excessive
The conclusion of activation.
The Chinese medicinal compound extract be able to can prepared with overexpression caused after specific downregulation ras Oncogene Mutations
Treatment is applied in the associated malignancies medicine as caused by being overexpressed after ras Oncogene Mutations, there is provided a kind of target is clear and definite
Treatment tumour medicine.
Embodiment
Specific embodiment presented below illustrates the present invention, but is not limited to these embodiments.
The preparation of one Chinese medicine compound prescription of embodiment
Material：Ginseng, ophiopogon are purchased from Lanzhou the Yellow River market.
(1) preparation of Chinese medicine compound prescription water decoction
Weigh ginseng 900g, ophiopogon 450g；Decocting, obtains total medicinal extract, is dissolved in water, constant volume, finally by gained sample preparation into
1.25mg/ml, 2.5mg/ml, 5mg/ml solution for standby.
(2) preparation of Chinese medicinal compound extract
Weigh ginseng 900g, ophiopogon 450g；Add ethanol solution to soak, filter, be concentrated under reduced pressure to give after being heated to reflux
The total medicinal extract of Chinese medicine compound prescription, is dissolved in water by total medicinal extract, is settled to constant volume 100ml；Chromatographic column, pre- place are loaded with D101 macroreticular resins
Reason, it is spare.Take 20ml medicinal extract liquid, upper D101 types macroporous resin column, respectively with 0%, 30%, 50%, 70%, ethanol solution
Elution, 10 times of column volumes rinse.
Two Chinese medicinal compound extract of embodiment improves the effect of Chinese medicine compound prescription water decoction toxicity
Ras is a kind of small molecular protein, participates in many important steps during regulation and control animal development.Ras signal paths
RAS albumen highly conserved from nematode to people, that let-60 gene codes are guarded in Caenorhabditis elegans, its RAS with the mankind
Albumen is compared with 83% homology.If ras is mutated, nematode can be caused to produce more vaginal orifices in Caenorhabditis elegans,
It is then the malignant proliferation for causing cell in the mankind, leads oncogenic generation.Therefore, model organism Caenorhabditis elegans
(Caenorhabditis elegans) Ras/MAPK signal paths excessive activation type mutant extensively should as test population
Suppress the antitumor drug of proto-oncogene ras path excessive activations for screening.
MT2124 is the Caenorhabditis elegans of let-60/ras excessive activations, can produce the phenotype of more vaginal orifices.Chinese medicine is answered
Side dilutes certain concentration, acts on Caenorhabditis elegans MT2124, if it is possible to suppresses the more vaginal orifices of tested nematode population and produces
The ratio of individual, and the wild type nematode individual ratio of only one Kidney-Yin door rises, then illustrate drug effect in ras paths,
The excessive activation of the path has been lowered, there is antitumor activity.The ratio of wild type nematode is higher in colony, then drug effect is imitated
Fruit is more notable.
1. experiment material
Caenorhabditis elegans strain：Caenorhabditis elegans mutant MT2124, genotype are：let-60(n1046sd,gf)
IV, purchased from CGC (Caenorhabditis Genetics Center)；
Bacterial strain：Uracil leaky mutant Escherichia coli OP50, as the food of Caenorhabditis elegans, buys in CGC.
2. the preparation of reagent
NGM culture mediums：NaCl1.5g, K2HPO41.3g, KH2PO48.5g, peptone 1.4g, agar powder 8.5g, supplement are steamed
Distilled water is to 500mL.The 500 μ L cholesterol (5mg/mL, absolute ethyl alcohol are prepared) of filtration sterilization, high pressure steam sterilization are added after sterilizing
500 μ L1mol/L MgSO4, 500 μ L1mol/L CaCl2。
LB fluid nutrient mediums：NaCl1g, peptone 1g, yeast extract 0.5g, supplement distilled water to 100mL.1M is used after dissolving
It is 7.0 that NaOH, which adjusts pH value, high-temperature sterilization.
M9 buffer solutions：Na2HPO4·12H2O15.12g, KH2PO43g, NaCl5g, MgSO4·7H2O0.25g, high temperature go out
Bacterium.
S bases formula of liquid (1L)：NaCl58.5g, K2HPO410g, KH2PO46g；Supplement distilled water to 1000mL, high temperature goes out
Bacterium.
1M potassium citrates (1L)：Citric acid H2O20g, potassium citrate H2O293.5g, supplement distilled water to 1000mL,
High-temperature sterilization.
Trace(1L)：Sodium-EDTA1.86g, FeSO4·7H2O0.69g, MnCl2·4H2O0.2g CuSO4·
5H2O0.29g, ZnSO4·7H2O0.025g, supplement distilled water to 1000mL, high-temperature sterilization.
S formula of liquid (1L)：S basal liquids 1L, 1M potassium citrate 10mL, Trace10mL, 1M CaCl23mL, 1M
MgSO43mL, 5mg/mL cholesterol 1mL, supplement distilled water to 1L, prepared before use.
3. instrument
High-pressure sterilizing pot (Shenan Medical Appliances Factory, Shanghai)
Single single side clean work station (Suzhou Decontamination Equipment Plant)
The intelligent biochemical cultivation cases of SPX (Ningbo Jiangnan instrument plant)
Constant-temperature shaking incubator (the permanent Science and Technology Ltd. in Shanghai one)
Water isolation type constant incubator (Shanghai Yiheng Scientific Instruments Co., Ltd)
Zoom-stereo microscope (Shanghai Yiheng Scientific Instruments Co., Ltd)
Electronic balance (Beijing Sai Duolisi instrument systems Co., Ltd）
Whirlpool blender (upper Nereid's tech device Co., Ltd)
Centrifuge (CT15E, Hitachi)
4. specific test procedure
The culture of nematode：Nematode is connected on the solid NGM plates for scribbling Escherichia coli OP50, then will be placed on 20 DEG C of training
Support and cultivated in case, synchronizing is handled when nematode grows to adult.
Nematode synchronizes：The nematode on tablet is swept away flushing liquor is sucked centrifuge tube with M9 buffer solutions；4000rpm/
Min centrifuges 3min, removes supernatant；Add lysate (final concentration：3.2%NaClO and 1M NaOH)；Shaken in vortex instrument
7min, makes the polypide of nematode rupture release ovum；4000rpm/min centrifuges 3min, removes supernatant, collects precipitation and obtains substantial amounts of line
Worm's ovum；M9 wash buffers twice, add 1ml M9 buffer solutions, be placed in 20 DEG C of incubators hatching 48 it is small when.
Drug effect：This experiment carries out on 96 orifice plates, and each processing sets five repetitions.First obtained synchronized
L1 phase nematode concentration adjust to the μ L of 80-100 bars/20.Then 140 μ L S liquid, 20 μ L liquids, 20 μ L nematodes are added into every hole
Liquid, 20 μ L OP50.It is put into 20 DEG C of constant-temperature shaking incubators and cultivates.After culture to adult, observed under inverted microscope,
Calculate WT ratio (%)=100% × wild type nematode number/(wild type nematode number+mutant nematode number).
5. Chinese medicine compound prescription water decoction suppresses ras path excessive activation bioactivity and its to Caenorhabditis elegans mutant
The influence of MT2124 growths
1 Chinese medicine compound prescription water decoction of table suppresses ras path excessive activation biological activity determinations and its dashes forward to Caenorhabditis elegans
The influence of variation MT2124 growths
Table 1 shows Chinese medicine compound prescription water decoction, can act on Caenorhabditis elegans ras mutant MT2124, significantly inhibit
More vaginal orifice phenotypes of MT2124, dramatically increase the WT ratios of Caenorhabditis elegans MT2124, and are in concentration-dependent relation,
Illustrate that Chinese medicine compound prescription water decoction has ras proto-oncogene excessive activations obvious downward to act on, mutant is changed into wild type.Separately
Outside, by nemic death rate it can be seen that Chinese medicine compound prescription water decoction shows Caenorhabditis elegans toxicity, and with concentration
Increase toxicity increase.
6. Chinese medicinal compound extract suppresses ras path excessive activation biological activity determinations and its Caenorhabditis elegans is mutated
The influence of body MT2124 growths
2 Chinese medicinal compound extract of table suppresses ras path excessive activation biological activity determinations and its dashes forward to Caenorhabditis elegans
The influence of variation MT2124 growths
After table 2 is the result shows that isolate and purify Chinese medicinal compound extract using D101 pillars, the elution of D101 pillars
Active ingredient focuses mostly on elutes section in ethanol solution, can dramatically increase the WT ratios of Caenorhabditis elegans MT2124
(P ﹤ 0.05), 77% is risen to by the 5% of blank control group, and active ingredient hardly shows toxicity.
Consolidated statement 1, the result of table 2, which can be seen that Chinese medicine compound prescription water decoction and Chinese medicinal compound extract, can significantly inhibit ras
Path excessive activation, improves the WT ratios of nematode, and still, the toxicity of Chinese medicine compound prescription water decoction is big, causes nematode largely dead
Die, and the present invention Chinese medicinal compound extract retain Chinese medicine compound prescription water decoction suppress ras path excessive activation effects while,
Toxicity is greatly reduced, and hardly causes nematode dead.Therefore, compared to the former, Chinese medicinal compound extract toxicity of the invention is more
Small, effect is more preferable.
Effect of the example IV Chinese medicinal compound extract to Caenorhabditis elegans MT8189 (lin-15 mutant inactives)
Experiment material：Caenorhabditis elegans MT8189, more vaginal orifice phenotypes, genotype:Lin-15b (n765) X, purchased from CGC
(Caenorhabditis Genetics Center)；Escherichia coli OP50, uracil leaky mutant, as beautiful hidden bar line
The food of worm, purchased from CGC.
Specific experiment step：With embodiment two.
It is located at the upstream of ras in lin-15 (n765) X, suppresses the expression of ras genes, the strain of MT8189 nematodes under normal circumstances
Gene inactivation causes nematode to be in more oopod phenotypes in system.
Mutant ratio of the present embodiment Chinese medicinal compound extract to Caenorhabditis elegans MT8189 (lin-15 mutant inactives)
Example ratio is 100%, illustrates that Chinese medicinal compound extract cannot suppress more vaginal orifice phenotypes of lin-15 (n765) X, according to genetically
The principle of position, since more cloudy caused by the reversible let60/ras proto-oncogene excessive activations downstream of Chinese medicinal compound extract
Door phenotype, then also suppress more vaginal orifice phenotypes of MT8189, but do not observe expected results in this experiment, i.e., Chinese medicine compound prescription is extracted
Thing cannot suppress more vaginal orifice phenotypes of let60/ras upstream genes lin-15 (n765) X.This is probably due to lin-15 (n765)
What is included in x-ray worm is wild type let60/ras copies, and Chinese medicinal compound extract only acts as the let60/ras of overexpression
With insensitive to wild type let60/ras, i.e., Chinese medicinal compound extract acts only on the RAS albumen of excessive activation, to wild type
RAS albumen is insensitive, this illustrates that what the Chinese medicinal compound extract only acted on ras excessive activations causes tumour cell, without
Act on normal cell.
Effect of five Chinese medicinal compound extract of embodiment to Caenorhabditis elegans CB1275 (lin-1 mutant inactives)
Experiment material：Caenorhabditis elegans genotype：Lin-1 (e1275) IV, purchased from CGC (Caenorhabditis
Genetics Center)；Escherichia coli OP50, uracil leaky mutant, as the food of Caenorhabditis elegans, is purchased from
CGC。
Specific experiment step：With embodiment two.
Lin-1 is a gene in ras downstreams, and it can be the negative of vaginal orifice development by the nuclear factor of MAPK phosphorylations to be
Regulatory factor, adjusts the signal path of the vaginal orifice development in let-60 downstreams.After lin-1 mutant inactives, nematode is more vaginal orifice phenotypes.
Since lin-1 is nuclear factor, the process of vaginal orifice cell development is adjusted.If medicine is to more vaginal orifice tables of the gene Inactivating mutations
Type has significant inhibitory action, then illustrates that medicine can directly suppress the development of vaginal orifice precursor, prevent its form more vaginal orifices and
In non-specific cytotoxicity.
The present embodiment Chinese medicinal compound extract is to the mutation rate of Caenorhabditis elegans CB1275 (lin-1 mutant inactives)
100%, illustrate that more vaginal orifice phenotypes of CB1275 strain nematodes are not affected by the influence of Chinese medicinal compound extract, this shows drug effect
The false vaginal orifice of ras excessive activations formation is can inhibit in ras signal paths, it is impossible to suppresses the false vaginal orifice of lin-1 mutant inactives formation
Phenotype, illustrates that Chinese medicinal compound extract acts on ras without acting on lin-1, it was demonstrated that medicine lowered the excessive activation of ras and
That the propagation for directly inhibiting cell works, i.e., it is that ras pathways rely on that Chinese medicinal compound extract, which works, rather than
Extensive cytotoxicity.
Shown by above example, Chinese medicine compound prescription can significantly be significantly inhibited Caenorhabditis elegans ras and be dashed forward by decocting
More vaginal orifice phenotypes of variation MT2124, dramatically increase the WT ratios of Caenorhabditis elegans MT2124, and are in concentration dependant
Relation, illustrates that Chinese medicine compound prescription has ras proto-oncogene excessive activations obvious downward to act on, mutant is changed into wild type.But
Decocting is only used, Chinese medicine compound prescription can increase Caenorhabditis elegans toxicity with the increase of concentration.The present invention provides a kind of Chinese medicine
The preparation process of compound extract, after being isolated and purified using D101 pillars to Chinese medicinal compound extract, Chinese medicine compound prescription extraction
The active ingredient of thing focuses mostly on elutes section in ethanol solution, the WT ratios of Caenorhabditis elegans MT2124 is significantly increased
Add, and active ingredient hardly shows toxicity.Present invention also offers the new application of Chinese medicinal compound extract, can suppress ras
Proto-oncogene is applied in being overexpressed and applied in the antitumor drug for suppressing to be overexpressed after ras Oncogene Mutations is prepared, and is had
Hope exploitation into the antitumor drug of a new generation.
Claims (3)
1. a kind of Chinese medicinal compound extract, it is characterised in that the Chinese medicine compound prescription is made of ginseng, ophiopogon, Chinese medicine compound prescription extraction
Thing is prepared by following method：In mass ratio 9:4.5 take ginseng, ophiopogon, add ethanol solution, and immersion, is heated to reflux,
Filtering, is concentrated under reduced pressure, obtains total medicinal extract；
Total medicinal extract is completely dissolved with distilled water, upper D101 types macroporous resin column, 24h is adsorbed, afterwards with 10 times of column volumes 50%
Ethanol eluate elution, eluent is concentrated into thick paste, dry, to obtain the final product.
2. application of the Chinese medicinal compound extract as claimed in claim 1 in antitumor drug is prepared, the tumour be by
Caused by being overexpressed after ras Oncogene Mutations.
3. Chinese medicinal compound extract as claimed in claim 1 is preparing the application in suppressing ras proto-oncogenes overexpression medicine.
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