CN105267619A - Antitumor traditional Chinese medicine compound extract for inhibiting ras proto-oncogene over-expression - Google Patents

Abstract

The invention relates to a Chinese medicine compound extract, a preparation method and application thereof. The Chinese medicine compound is composed of ginseng and Ophiopogon japonicus. Studies have shown that about 90% of pancreatic cancer, 50% of colon cancer, 30%-40% of lung adenocarcinoma and 5%-40% of leukemia are caused by the overexpression of ras gene, which has become a recognized screening anti-cancer drug target. The invention discloses a preparation process of the compound Chinese medicine extract. Compared with its water decoction treatment solution, the compound Chinese medicine extract prepared by the process has lower toxicity and better effect of inhibiting excessive activation of ras gene pathway. In addition, the present invention also provides a new application of a traditional Chinese medicine compound extract, which can be used in inhibiting the overexpression of the ras proto-oncogene and in the preparation of anti-tumor drugs for inhibiting the overexpression of the ras proto-oncogene after mutation.

Description

An anti-tumor traditional Chinese medicine compound extract that inhibits the overexpression of ras proto-oncogene

technical field

The invention relates to a compound Chinese medicine extract and a preparation process thereof. Specifically, the Chinese medicine compound is composed of ginseng and Ophiopogon japonicus, and specifically relates to the application of the Chinese medicine compound extract in inhibiting the overexpression of the ras proto-oncogene and in the preparation of the anti-ras proto-oncogene The invention relates to the application of antitumor drugs overexpressed after cancer gene mutation, belonging to the field of antitumor drugs.

Background technique

At present, malignant tumors have become the number one killer of human beings, seriously endangering human health. According to the data of the World Health Organization, the incidence of tumors in the world is 10 million people every year, and about 7 million people die. The number of cases is 2 million, and the death due to cancer is about 1.4 million. Among the 5 people who die in our country, 1 person dies from cancer.

About 30% of human tumors are caused by the activation of ras proto-oncogene mutation, which leads to the increased expression of RAS protein. If the RAS protein is continuously activated, it can bind to the downstream effector protein and transmit the signal to the downstream signaling element, causing abnormal proliferation of cells and leading to the occurrence of tumors. Anti-tumor therapy targeting the ras signaling pathway has become a tumor therapy. Therefore, the ras proto-oncogene has become a recognized target for screening drugs against related malignant tumors.

RAS protein plays an important role in regulating several important signaling pathways involved in the regulation of normal cell proliferation. Therefore, tumor therapy targeting RAS protein and the signaling pathway regulated by RAS protein is effective in the treatment of tumors caused by ras proto-oncogene mutations. It is of great value in tumors. On December 20, 2005, Nexavar, a drug for the treatment of advanced renal cell carcinoma approved by the US FDA, directly targets the Ras signaling pathway, which can inhibit the Raf/MEK/ERK signaling pathway and directly inhibit tumor growth; the drug targeting the ras effector pathway Kinase inhibitors such as ID1839 and OSI774, which can effectively block the activity of EGFR tyrosine kinase and to some extent also inhibit other members of the ERBB protein family, but these are not drugs that directly target ras. Farnesylation transferase inhibitors targeting ras protein can also inhibit farnesylation of most other proteins (possibly including some normal human proteins), but there is also the problem of poor specificity. Therefore, the development of safe and efficient therapeutic Antitumor drugs that overactivate the ras proto-oncogene are urgently needed.

Our country's traditional Chinese medicine has achieved certain curative effect in clinical tumor treatment, and has low toxicity, which has opened up a new way for the prevention and treatment of tumor.

Ginseng is the dried root and rhizome of Panaxginseng C.A.Mey, the main active ingredient of which is ginsenoside and ginseng polysaccharide. Ginseng has the functions of invigorating vital energy, nourishing the spleen and benefiting the lung, promoting body fluid and calming the mind, anti-fatigue, anti-tumor, anti-shock, and lowering blood fat.

Ophiopogon japonicus (Thumb) Ker-Gawl is the dry root tuber of the perennial herb of Liliaceae Ophiopogon japonicus (Thumb) Ker-Gawl. Ophiopogon japonicus is slightly cold in nature, sweet in taste, slightly bitter, and has the functions of nourishing yin and nourishing essence, nourishing yin and qi, clearing the heart and eliminating troubles, moistening the intestines and laxative, etc. It is one of the commonly used traditional Chinese medicines. Ophiopogon japonicus has a wide range of pharmacological effects, such as anti-myocardial ischemia, anti-arrhythmia, anti-aging, anti-tumor, immune regulation and so on.

So far, no literature has reported that a Chinese medicine compound containing ginseng and Maidong can inhibit the overexpression of ras proto-oncogene after mutation, and there is no Chinese medicine compound extract containing ginseng and Maidong as raw materials on the market that can specifically inhibit Antitumor drugs caused by overexpression of ras proto-oncogene mutation. However, according to the characteristics of ginseng and Ophiopogon japonicus, through hard work and a large number of screening analysis, the present invention has developed a Chinese medicine compound for the treatment of tumors with excessive activation of the ras proto-oncogene. , achieve the purpose of treating tumors, and have low toxicity and no teratogenic effect, and are suitable for use in the preparation of related anti-cancer drugs caused by the overexpression of the ras proto-oncogene mutation, and can be combined with any pharmaceutically acceptable The auxiliary materials are mixed to make various dosage forms, which are expected to be developed into a new generation of anti-tumor drugs.

In the process of preparing the Chinese medicine compound extract, it was found that only water decoction treatment can significantly inhibit the excessive activation of the ras gene pathway, but the toxicity is high and the safety is poor. Therefore, the invention discloses a preparation process of the Chinese medicine compound extract, in which the compound is reflux extracted with absolute ethanol, and on the basis of alcohol extraction, it is further separated and purified through a macroporous resin, and after separation and purification The composition of the decoction not only retains the effect of the water decoction on inhibiting the overexpression of the ras proto-oncogene, but also greatly reduces the toxicity of the traditional Chinese medicine compound water decoction, so the effect is better. application in medicines.

Contents of the invention

The purpose of the present invention is to provide a preparation process of a compound Chinese medicine extract. Specifically, the compound Chinese medicine is composed of ginseng and Ophiopogon japonicus, in order to provide a compound Chinese medicine extract with high antitumor activity, low toxicity, and low cost. The present invention Another purpose of the present invention is to provide a new application of a Chinese medicine compound extract, specifically related to the application of the Chinese medicine compound extract in inhibiting the overexpression of the ras proto-oncogene and the preparation of an anti-tumor drug that inhibits the overexpression of the ras proto-oncogene after mutation in the application.

The present invention is implemented through the following technical solutions.

A kind of Chinese medicine compound extract according to the present invention, wherein the Chinese medicine compound is composed of ginseng and Ophiopogon japonicus, and the Chinese medicine compound extract is prepared by the following method: take ginseng and Ophiopogon japonicus, add ethanol solution, soak, heat and reflux, Filtrate, concentrate under reduced pressure to obtain the total extract; completely dissolve the total extract with distilled water, put it on a macroporous resin column, absorb for 24 hours, and then elute with the eluent, concentrate the eluent to a thick extract, and dry to obtain.

The traditional Chinese medicine compound extract of the present invention is made from ginseng and Ophiopogon japonicus as raw materials, and the mass ratio of the raw materials is 4.5-9:4.5-9.

The traditional Chinese medicine compound extract of the present invention is made from ginseng and Ophiopogon japonicus as raw materials, and the mass ratio of the raw materials is 9:4.5.

In the Chinese medicine compound extract of the present invention, the ethanol solution during the reflux extraction is absolute ethanol.

In the Chinese medicine compound extract of the present invention, the macroporous resin is D101 type macroporous resin.

According to the Chinese medicine compound extract of the present invention, the elution volume of the total Chinese medicine compound extract is 10 times of column volume when it is separated, purified and eluted.

According to the Chinese medicine compound extract of the present invention, the eluent used in the separation and purification of the total extract of the Chinese medicine extract is ethanol solution.

In the Chinese medicine compound extract described in the present invention, the ethanol solution used for eluting the total extract of the Chinese medicine extract during separation and purification is absolute ethanol. At this time, while ensuring the efficacy of the Chinese medicine compound extract, it hardly shows toxicity.

The invention is a traditional Chinese medicine compound extract, and the said tumor is caused by overexpression after ras proto-oncogene mutation.

The invention relates to the application of the compound extract of traditional Chinese medicine in inhibiting the overexpression of ras proto-oncogene.

For the application of the compound Chinese medicine extract of the present invention in the preparation of anti-tumor drugs, the compound Chinese medicine extract can be added with pharmaceutically acceptable carriers such as fillers, lubricants, wetting agents, absorption-promoting agents, adsorption carriers, diluents, etc. Agents, excipients, etc., sweeteners, flavoring agents, etc. can be added if necessary, and prepared into various dosage forms of injections, powders, granules, powders, pills, oral liquids, tablets, etc. Those skilled in the art understandable.

Beneficial effects: the present invention provides a traditional Chinese medicine compound extract for treating tumors with overexpression of the ras proto-oncogene and its application in the preparation of related malignant tumor drugs. Compared with the existing technology, it has the following advantages:

(1) The Chinese medicine compound extract prepared by the preparation process of the present invention has good effect, can significantly reduce toxicity, and reduces the mortality rate of the model Caenorhabditis elegans, which inhibits the overactivation of the ras proto-oncogene pathway, and increases the proportion of wild type. high.

(2) The traditional Chinese medicine compound extract prepared by the preparation process of the present invention can significantly inhibit the overexpression of the ras proto-oncogene, and can be used in the preparation and treatment of tumor drugs caused by the overexpression of the ras proto-oncogene.

(3) The formula ratio between ginseng and Ophiopogon japonicus in the traditional Chinese medicine compound can be rationally changed according to the principle of traditional Chinese medicine formula, and the Chinese medicine compound extract prepared by the preparation process of the present invention can still significantly down-regulate the excessive activation of the ras proto-oncogene , has antitumor activity.

(4) The traditional Chinese medicine compound extract prepared by the preparation process of the present invention only acts on the overexpressed mutant RAS protein, but has no effect on the wild-type RAS protein. It is suggested that the compound extract of traditional Chinese medicine is safe in application, and it only acts on cancer cells, but not on normal cells. In the future, we are expected to apply the compound extract of traditional Chinese medicine in the preparation of anti-related malignant tumor drugs, wherein the tumor is caused by the overexpression of ras proto-oncogene, so as to provide better treatment for patients with such tumors, and provide The treatment of some difficult cancers provides new avenues.

(5) The compound extract of traditional Chinese medicine significantly down-regulates the overexpression of ras proto-oncogene, which is not caused by mutagenesis and does not cause teratogenicity.

(6) Since the ras signaling pathway is highly conserved from nematodes to humans, the let-60 gene encodes a conserved RAS protein in Caenorhabditis elegans, which has 83% homology with human RAS proteins. If the ras mutation, in Caenorhabditis elegans, it will cause the nematode to produce polyvagina, and in humans, it will cause the malignant proliferation of cells, leading to the occurrence of tumors. Therefore, the model organism Caenorhabditis elegans (Caenorhabditis legans) Ras/MAPK signaling pathway hyperactivation mutants are widely used as a test group to screen anti-tumor drugs that inhibit the hyperactivation of the ras proto-oncogene pathway. Therefore, the present invention adopts the Caenorhabditis elegans mutant strain as a model for evaluating drugs that inhibit the overactivation of the ras proto-oncogene pathway, and obtains that the compound extract of traditional Chinese medicine has a significant down-regulation effect on the overexpression of the ras proto-oncogene, and has the ability to inhibit the ras proto-oncogene pathway. Conclusions for overactivation of oncogene pathways.

The traditional Chinese medicine compound extract can specifically down-regulate the overexpression caused by the mutation of the ras proto-oncogene, and can be used in the preparation and treatment of related malignant tumor drugs caused by the overexpression of the ras proto-oncogene mutation, providing a specific target Drugs to treat tumors.

detailed description

Specific examples are provided below to illustrate the present invention, but are not limited to these examples.

The preparation of embodiment one Chinese medicine compound

Materials: Ginseng and Ophiopogon japonicus were purchased from the Yellow River Market in Lanzhou.

(1) Preparation of Chinese medicine compound water decoction

Weigh 900g of ginseng and 450g of Ophiopogon japonicus; decoct in water to obtain the total extract, dissolve in water, constant volume, and finally prepare the obtained samples into 1.25mg/ml, 2.5mg/ml, 5mg/ml solutions for later use.

(2) Preparation of Chinese medicine compound extract

Weigh 900g of ginseng and 450g of Radix Ophiopogon japonicus; soak in absolute ethanol solution, filter after heating and reflux, and concentrate under reduced pressure to obtain the total Chinese medicine compound extract, dissolve the total extract in water, and set the volume to 100ml; use D101 macroporous The chromatographic column is filled with resin, pretreated and set aside. Take 20ml of the extract, put it on a D101 macroporous resin column, elute with 0%, 30%, 50%, 70%, and absolute ethanol solution, and wash with 10 times the column volume.

Embodiment two Chinese medicine compound extracts improve the effect of Chinese medicine compound water decoction toxicity

Ras is a small molecular protein that participates in the regulation of many important links in the development of animals. The ras signaling pathway is highly conserved from nematodes to humans. In Caenorhabditis elegans let-60 gene encodes a conserved RAS protein, which has 83% homology with human RAS protein. If the ras mutation, in Caenorhabditis elegans, it will cause the nematode to produce polyvagina, and in humans, it will cause the malignant proliferation of cells, leading to the occurrence of tumors. Therefore, the model organism Caenorhabditis elegans (Caenorhabditiselegans) Ras/MAPK signaling pathway overactivation mutants are widely used as a test group to screen anti-tumor drugs that inhibit the overactivation of proto-oncogene ras pathway.

MT2124 is a let-60/ras overactivated C. elegans that produces a polyvaginal phenotype. Dilute the traditional Chinese medicine compound to a certain concentration and act on Caenorhabditis elegans MT2124. If the proportion of individuals with multiple vulva in the tested nematode population can be suppressed, while the proportion of wild-type nematode individuals with only one true vulva increases, it means that the drug acts on the ras pathway , which down-regulates the overactivation of this pathway and has antitumor activity. The higher the proportion of wild-type nematodes in the population, the more pronounced the effect of the drug.

1. Experimental materials

Caenorhabditis elegans strain: Caenorhabditis elegans mutant MT2124, genotype: let-60(n1046sd, gf) IV, purchased from CGC (Caenorhabditis Genetics Center);

Bacterial strain: uracil-leaking mutant E. coli OP50, used as food for C. elegans, purchased from CGC.

2. Preparation of reagents

NGM medium: NaCl 1.5g, K ₂ HPO ₄ 1.3g, KH ₂ PO ₄ 8.5g, peptone 1.4g, agar powder 8.5g, add distilled water to 500mL. After sterilization, 500 μL of cholesterol (5 mg/mL, prepared in absolute ethanol) sterilized by filtration, 500 μL of 1mol/LMgSO ₄ and 500 μL of 1mol/LCaCl ₂ sterilized by high pressure steam were added.

LB liquid medium: NaCl 1g, peptone 1g, yeast extract 0.5g, add distilled water to 100mL. After dissolving, adjust the pH value to 7.0 with 1M NaOH, and sterilize at high temperature.

M9 buffer solution: Na ₂ HPO ₄ ·12H ₂ O 15.12g, KH ₂ PO ₄ 3g, NaCl 5g, MgSO ₄ ·7H ₂ O 0.25g, high temperature sterilization.

S base solution formula (1L): 58.5g NaCl, 10g K ₂ HPO ₄ , 6g KH ₂ PO ₄ ; add distilled water to 1000mL, and sterilize at high temperature.

1M potassium citrate (1L): Citric acid·H ₂ O 20g, potassium citrate·H ₂ O 293.5g, add distilled water to 1000mL, and sterilize at high temperature.

Trace (1L): Sodium-EDTA 1.86g, FeSO ₄ 7H ₂ O 0.69g, MnCl ₂ 4H ₂ O 0.2g CuSO ₄ 5H ₂ O 0.29g, ZnSO ₄ 7H ₂ O 0.025g, add distilled water to 1000mL, High temperature sterilization.

S solution formula (1L): S base solution 1L, 1M potassium citrate 10mL, Trace 10mL, 1MCaCl ₂ 3mL, 1MMgSO ₄ 3mL, 5mg/mL cholesterol 1mL, add distilled water to 1L, and prepare before use.

3. Instrument

Autoclave (Shanghai Shen'an Medical Instrument Factory)

Single-person single-sided purification workbench (Suzhou Purification Equipment Factory)

SPX intelligent biochemical incubator (Ningbo Jiangnan Instrument Factory)

Constant temperature culture shaker (Shanghai Yiheng Technology Co., Ltd.)

(Shanghai Yiheng Scientific Instrument Co., Ltd.

Water-proof constant temperature incubator)

(Shanghai Yiheng Scientific Instrument Co., Ltd.

Continuous Zoom Stereo Microscope Division)

(Beijing Sartorius Instrument System has

Electronic Balance Co., Ltd.)

Vortex mixer (Shanghai Jingke Instrument Co., Ltd.)

Centrifuge (CT15E, Hitachi)

4. Specific test steps

Cultivation of nematodes: connect nematodes to solid NGM plates coated with Escherichia coli OP50, and then culture them in an incubator at 20°C, and perform synchronization when the nematodes grow into adults.

Synchronization of nematodes: wash down the nematodes on the plate with M9 buffer solution, suck the washing solution into a centrifuge tube; centrifuge at 4000rpm/min for 3min, remove the supernatant; add lysate (final concentration: 3.2% NaClO and 1MNaOH); Shake on the instrument for 7 minutes to rupture the nematodes and release the eggs; centrifuge at 4000rpm/min for 3 minutes, remove the supernatant, collect the precipitate to obtain a large number of nematode eggs; wash twice with M9 buffer, add 1ml of M9 buffer, and incubate at 20°C Incubated in the box for 48 hours.

Drug effect: This experiment was carried out on a 96-well plate, with five replicates for each treatment. First adjust the concentration of nematodes in the L1 phase obtained through synchronization to 80-100 nematodes/20 μL. Then add 140 μL of L solution, 20 μL of drug solution, 20 μL of nematode solution, and 20 μL of OP50 into each well. Place them in a constant temperature culture shaker at 20°C. After culturing to adults, observe under an inverted microscope, and calculate the wild-type ratio (%)=100%×number of wild-type nematodes/(number of wild-type nematodes+number of mutant nematodes).

5. Traditional Chinese medicine compound water decoction inhibits the biological activity of ras pathway overactivation and its effect on the growth of Caenorhabditis elegans mutant MT2124

Table 1 Determination of biological activity of traditional Chinese medicine compound water decoction in inhibiting excessive activation of ras pathway and its effect on the growth of Caenorhabditis elegans mutant MT2124

Table 1 shows that the Chinese medicine compound decoction can act on the Caenorhabditis elegans ras mutant MT2124, significantly inhibit the polyvulva phenotype of MT2124, and significantly increase the wild-type ratio of Caenorhabditis elegans MT2124 in a concentration-dependent manner, indicating Chinese medicine compound water decoction can significantly down-regulate the overactivation of ras proto-oncogene, making the mutant become wild type. In addition, from the nematode mortality rate, it can be seen that the Chinese medicine compound water decoction shows toxicity to Caenorhabditis elegans, and the toxicity increases with the increase of the concentration.

6. Determination of biological activity of Chinese medicine compound extracts inhibiting excessive activation of ras pathway and its effect on the growth of Caenorhabditis elegans mutant MT2124

Table 2 Determination of biological activity of traditional Chinese medicine compound extracts in inhibiting excessive activation of ras pathway and its effect on the growth of Caenorhabditis elegans mutant MT2124

The results in Table 2 show that after using the D101 column to separate and purify the Chinese medicine compound extract, the effective components eluted from the D101 column are mostly concentrated in the ethanol solution elution section, which can significantly increase the wild-type ratio of Caenorhabditis elegans MT2124 (P <0.05), from 5% of the blank control group to 77%, and the active ingredients hardly show toxicity.

From the results of Table 1 and Table 2, it can be seen that the Chinese medicine compound decoction and the Chinese medicine compound extract can significantly inhibit the excessive activation of the ras pathway and increase the wild-type ratio of nematodes. However, the Chinese medicine compound decoction is highly toxic and causes nematodes A large number of nematodes died, while the traditional Chinese medicine compound extract of the present invention retains the effect of the traditional Chinese medicine compound water decoction on inhibiting the excessive activation of the ras pathway, while the toxicity is greatly reduced, and it hardly causes nematodes to die. Therefore, compared with the former, the traditional Chinese medicine compound extract of the present invention has less toxicity and better effect.

Example 4 The effect of Chinese medicine compound extract on Caenorhabditis elegans MT8189 (lin-15 mutation inactivation)

Experimental materials: Caenorhabditis elegans MT8189, polyvulva phenotype, genotype: lin-15b(n765)X, purchased from CGC (Caenorhabditis Genetics Center); Escherichia coli OP50, a uracil-leaking mutant strain, used as food for Caenorhabditis elegans , purchased from CGC.

Concrete experimental procedure: with embodiment two.

lin-15(n765)X is located upstream of ras, and normally inhibits the expression of ras gene. Inactivation of this gene in the MT8189 nematode strain leads to the phenotype of multiple ovipositors in nematodes.

In this example, the ratio of the compound Chinese medicine extract to the mutant of Caenorhabditis elegans MT8189 (lin-15 mutation inactivation) was 100%, indicating that the compound Chinese medicine extract could not inhibit the polyvulva phenotype of lin-15(n765)X, According to the principle of genetic epistasis, since the compound extract of traditional Chinese medicine can reverse the polyvulva phenotype caused by the overactivation of its downstream let60/ras proto-oncogene, it can also inhibit the polyvulva phenotype of MT8189, but no expected results were observed in this experiment , that is, the compound extract of traditional Chinese medicine can not inhibit the polyvulva phenotype of let60/ras upstream gene lin-15(n765)X. This may be due to the wild-type let60/ras copy contained in lin-15(n765) X. The extract only acts on overactivated RAS protein, and is insensitive to wild-type RAS protein, which indicates that the Chinese medicine compound extract only acts on tumor cells that cause excessive ras activation, but not on normal cells.

Example 5 Effects of Chinese Medicine Compound Extracts on Caenorhabditis elegans CB1275 (lin-1 mutation inactivation)

Experimental materials: Caenorhabditis elegans genotype: lin-1(e1275)IV, purchased from CGC (Caenorhabditis Genetics Center); Escherichia coli OP50, a uracil-leaking mutant strain, used as food for Caenorhabditis elegans, purchased from CGC.

Concrete experimental procedure: with embodiment two.

lin-1 is a gene downstream of ras, a nuclear transcription factor that can be phosphorylated by MAPK, a negative regulator of vulva development, and a signaling pathway that regulates vulva development downstream of let-60. After lin-1 mutation inactivation, nematodes have polyvaginal phenotype. Since lin-1 is a nuclear transcription factor, it regulates the process of vulva cell development. If the drug has a significant inhibitory effect on the polyvulva phenotype of the gene inactivation mutation, it means that the drug can directly inhibit the development of vulva precursor cells, prevent them from forming polyvulva and exhibit non-specific cytotoxicity.

The mutation rate of the Chinese medicine compound extract in this example to Caenorhabditis elegans CB1275 (lin-1 mutation inactivation) is 100%, indicating that the polyvulva phenotype of the CB1275 strain nematode is not affected by the Chinese medicine compound extract, which shows that the drug Acting on the ras signaling pathway can inhibit the false vulva formed by excessive activation of ras, but cannot inhibit the false vulva phenotype formed by lin-1 mutation inactivation, indicating that the compound Chinese medicine extract acts on ras but not on lin-1, proving that the drug down-regulates The overactivation of ras does not directly inhibit the proliferation of cells, that is, the effect of the compound extract of traditional Chinese medicine is dependent on the mechanism of ras pathway, rather than extensive cytotoxicity.

The above examples show that the traditional Chinese medicine compound can significantly inhibit the polyvaginal phenotype of the Caenorhabditis elegans ras mutant MT2124 by decocting in water, and significantly increase the wild-type ratio of the Caenorhabditis elegans MT2124 in a concentration-dependent manner, indicating that The traditional Chinese medicine compound can significantly down-regulate the overactivation of ras proto-oncogene, making the mutant become wild type. But only decocting with water, the Chinese medicine compound will be more toxic to C. elegans with the increase of concentration. The invention provides a preparation process of a compound Chinese medicine extract. After separating and purifying the compound Chinese medicine extract with a D101 column, the active ingredients of the compound Chinese medicine extract are mostly concentrated in the elution section of anhydrous ethanol solution, so that Caenorhabditis elegans The wild-type ratio of MT2124 was significantly increased, and the active ingredient showed little toxicity. The present invention also provides a new application of the Chinese medicine compound extract, which can be used in inhibiting the overexpression of the ras proto-oncogene and in the preparation of an antitumor drug that inhibits the overexpression of the ras proto-oncogene after mutation, and is expected to be developed into a new generation of anti-cancer drugs. tumor drugs.

Claims (10)

1. a Chinese medicine compound extract, it is characterized in that, this Chinese medicine compound is made up of Radix Ginseng, Ophiopogon japonicus, and this Chinese medicine compound extract is prepared by following method: get Radix Ginseng, Ophiopogon japonicus, add ethanol solution, soak, reflux, Filter and concentrate under reduced pressure to obtain the total extract; The total extract is completely dissolved in distilled water, put on a macroporous resin column, adsorbed for 24 hours, then eluted with an eluent, concentrated to a thick paste, dried, and obtained. 2. A compound Chinese medicine extract according to claim 1, wherein the compound Chinese medicine contains ginseng and Ophiopogon japonicus in a mass ratio of 4.5-9:4.5-9. 3. A compound Chinese medicine extract according to claim 1, characterized in that the compound Chinese medicine contains ginseng and Ophiopogon japonicus in a mass ratio of 9:4.5. 4. a kind of Chinese medicine compound extract according to claim 1, is characterized in that, described ethanol solution is dehydrated alcohol. 5. A compound Chinese medicine extract according to claim 1, wherein the macroporous resin is D101 type macroporous resin. 6. A kind of Chinese medicine compound extract according to claim 1, characterized in that, the elution amount during the separation and purification of the total extract of the Chinese medicine compound is 10 times the column volume. 7. A kind of Chinese medicine compound extract according to claim 1, characterized in that, the eluent used in the separation and purification of the total extract of the Chinese medicine extract is ethanol solution. 8. a kind of Chinese medicine compound extract according to claim 7, is characterized in that, described ethanol solution is dehydrated alcohol. 9. The application of the traditional Chinese medicine compound extract as claimed in claim 1-8 in the preparation of antitumor drugs, wherein the tumor is caused by the overexpression of ras proto-oncogene after mutation. 10. The application of the Chinese medicine compound extract as claimed in claims 1-8 in inhibiting the overexpression of ras proto-oncogene.