CN109512995B - Anti-tumor traditional Chinese medicine composition and volatile oil extract thereof - Google Patents
Anti-tumor traditional Chinese medicine composition and volatile oil extract thereof Download PDFInfo
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Abstract
The invention relates to a traditional Chinese medicine composition and application of volatile oil extract thereof in drugs for inhibiting ras proto-oncogene mutation and then overexpressing tumors. The traditional Chinese medicine composition and the volatile oil extract thereof can obviously inhibit tumor-like multiple-vulvar phenotype of caenorhabditis elegans with ras protooncogene overexpression caused by let-60 mutation, so that the caenorhabditis elegans is twisted into a wild-type phenotype with a true-vulvar. The results show that the traditional Chinese medicine composition and the volatile oil extract thereof provided by the invention have the function of treating ras gene overexpressed tumors and can be applied to preparation of anti-tumor medicines for inhibiting ras proto-oncogene mutation and overexpression.
Description
Technical Field
The invention relates to a traditional Chinese medicine composition and application of a volatile oil extract thereof in an antitumor drug for inhibiting ras proto-oncogene mutation and then performing overexpression, and belongs to the field of antitumor drugs.
Background
At present, malignant tumors become the first killers of human beings, which seriously harm the health of human beings, and the data of the world health organization show that the incidence of tumors worldwide is 1000 thousands of people each year, about 700 thousands of people die, the number of new cases of cancer in China is 200 thousands each year, and about 140 thousands of cancer dies, so that 1 of 5 people die of cancer in China (Wangming politics, 2017).
About 30% of human tumors are due to RAS proto-oncogene mutation leading to overactivation of RAS proteins. Mutations in the RAS gene and overactivation of RAS proteins occur in 30% of human tumors, with mutations in the RAS gene occurring in about 90% of pancreatic cancers, around 50% of colon cancers and around 20% of acute leukemias (Hara and Han, 1995). KRAS mutations were detected in 40% of hereditary nonpolyposis colorectal cancers, 49% of conventional follicular thyroid cancers have RAS mutations, and 48% of Hurthle cell thyroid tumors have RAS mutations. KRAS codon 12 mutations occur in 83% of pancreatic cancers and 85% of biliary cancers. In addition, cancers such as lung cancer, gastric cancer, breast cancer, bladder cancer, and melanoma are closely related to ras gene mutations (Yangqi Wei et al, 2018). Ras-MAPK signal transduction pathway abnormality is involved in the process of liver cancer. Anti-tumor therapy targeting Ras signaling pathway also becomes a research hotspot in tumor therapeutics, and therefore, Ras has become a well-known target for screening related malignant cancer drugs (liuxuemei et al, 2008). The Ras signaling pathway is highly conserved from nematode to human, and the let-60 gene encodes a conserved Ras protein in caenorhabditis elegans, which has 83% homology to human Ras proteins. If the ras gene is mutated, it causes multiple vulva production by caenorhabditis elegans and malignant proliferation of cells in humans, leading to tumorigenesis (Lepistil et al, 2016). Therefore, the model organism Caenorhabditis elegans (Caenorhabditis elegans) Ras/MAPK signal channel overactivation type mutant is widely applied to screening of antitumor drugs inhibiting Ras channel overactivation as a tumor model (Hara and Han, 1995; Wang 261212005; Liuxue Mei et al, 2008). The invention adopts caenorhabditis elegans as a screening tool to screen candidate drugs of the tumor drug with the over-activation of Ras/MAPK signal channels.
Traditional combination of chemical drug therapy with surgery, radiotherapy, etc. has successfully improved the treatment rate of various malignant tumors, but due to strong side effects and drug resistance, the research for finding new treatment tumors is still reluctant (Zhanguoyun, 2012). The Chinese medicine has the advantages of multiple aspects, multiple targets, difficult drug resistance generation and the like in treating cancers, accords with the mechanism of tumor multifactor and multi-link pathogenesis, has the research on the effective part, the effective components and the action mechanism of the anti-tumor, becomes a hot point of the current research at home and abroad, has certain curative effect in the aspect of clinical tumor treatment, has low toxicity, opens up a new path for tumor prevention and treatment, and in recent years, the anti-tumor research work of the Chinese medicine and the effective components thereof has a series of achievements, and some achievements have been converted into the anti-tumor drugs widely used in clinic (Yilong et al, 2006, Zhang Xiuyun, 2012, Zhu Yuan et al, 2017).
The Longhu Rendan is originally originated from the ancient Chinese patent 'Zhuge March san', and the existing prescription is obtained after a few improvements. The LONGHURENDAN is mainly prepared from Mentholum, Borneolum Syntheticum, flos Caryophylli, fructus Amomi, fructus Anisi Stellati, cortex Cinnamomi, fructus Piperis, radix aucklandiae, Zingiberis rhizoma, Catechu, and Glycyrrhrizae radix. Has effects in inducing resuscitation, refreshing mind, dispelling summerheat, eliminating turbid pathogen, regulating stomach, and relieving vomit, and can be used for treating dizziness due to heatstroke, nausea, emesis, diarrhea, car sickness, and seasickness. To date, there is no report in the literature that longhurendan has anti-tumor activity.
The invention is improved on the basis of the Longhurendan, and discovers a traditional Chinese medicine composition capable of inhibiting ras gene overactivation and a volatile oil extract thereof, wherein the traditional Chinese medicine composition and the volatile oil extract thereof can be applied to treatment of ras proto-oncogene overactivated tumors. Compared with the Longhurendan, the traditional Chinese medicine composition and the volatile oil extract thereof can be applied to treating tumors with ras proto-oncogene over-activation.
The traditional Chinese medicine composition and the volatile oil extract thereof provided by the invention can be mixed with any pharmaceutically acceptable auxiliary material to prepare various dosage forms, and can be applied to treatment of ras proto-oncogene overactivated tumors.
Reference to the literature
Wangming politics, Chinese malignant tumor prevention and control strategy exploration [ J ] Chinese medicine management journal, 2017,25(06):8-9
Liuxue plum, Deng Ping, etc., Ras signal transduction pathway-based drug target research progress [ J ] pharmaceutical progress, 2008,32(11): 481-485-
Queensland 26121, (2005) use of nematodes as model organisms to screen drugs that inhibit oncogene ras activity.
Hara M，Han M et al.Ras farnesyltransferase inhibitors suppress the phenotype resulting from an activated ras mutation in Caenorhabditis elegans.[J].Developmental Biology，1995,92(4):3333-3337
Zhangxiyun et al, research progress of effective parts and chemical components of anti-tumor Chinese medicine [ J ] scientific and technological View, 2012,7(3):177-
"Yilong, Xuliang, etc., the research status of the action of anti-tumor Chinese medicine and its effective ingredients [ J ] animal medicine progress 2006,27(1):39-43
Zhuyuanzhu, Zuibibusao, etc., research progress of anti-tumor mechanism of volatile oil of Chinese medicine [ J ]. J. Chinese Experimental formulary, 16(8):227-
The research progress of the mutation rate of Yangqi Wei, Suizhijie, Duzhenwu, Zhangzhen RAS gene family in solid tumor [ J ] Chinese stereology and image analysis, 2018(03):303-310.
The role of the plum stamen, Songweijiang, Zhanghao, Yaoquan, Ras-MAPK pathway in the pathogenesis of liver cancer [ J ]. Zhongri friendly Hospital's report, 2016,30(01):47-49+54.
Disclosure of Invention
The invention aims to provide a traditional Chinese medicine composition which is prepared from clove, amomum villosum, star anise, cinnamon, white pepper, elecampane and dried ginger through the steps of extracting by a steam distillation method to obtain a crude product of a volatile oil extract of the traditional Chinese medicine composition, extracting by using an organic solvent, volatilizing the solvent by extracting an obtained organic phase, dehydrating and drying to obtain the traditional Chinese medicine volatile oil extract. The invention provides application of the traditional Chinese medicine composition and volatile oil extract thereof in inhibition of ras proto-oncogene overexpression, and particularly relates to application of the traditional Chinese medicine composition and volatile oil extract thereof in inhibition of ras proto-oncogene overexpression, in particular to application of tumors in treatment of liver cancer, pancreatic cancer, lung cancer and colon cancer.
The invention is implemented by the following technical scheme.
The invention aims to provide a traditional Chinese medicine composition which is prepared from the following raw materials in parts by weight: 5 parts of clove, 5 parts of amomum villosum, 3 parts of star anise, 8 parts of cinnamon, 3 parts of white pepper, 3 parts of costustoot and 5 parts of dried ginger.
The invention also aims to provide a traditional Chinese medicine volatile oil extract which is prepared from 5 parts of clove, 5 parts of amomum villosum, 3 parts of star anise, 8 parts of cinnamon, 3 parts of white pepper, 3 parts of costus root and 5 parts of dried ginger, and the preparation method comprises the following steps: adding distilled water into the clove, the amomum villosum, the star anise, the cinnamon, the white pepper, the elecampane and the dried ginger according to the proportion, extracting by adopting a steam distillation method to obtain a crude volatile oil extract product, extracting the crude volatile oil extract product by using an organic solvent, volatilizing the solvent from an organic phase obtained by extraction, and dehydrating and drying to obtain the traditional Chinese medicine volatile oil extract.
The Chinese medicine volatile oil extract has the marker chemical components including eugenol 42.43-59.19%, eugenol acetate 13.05-15.28%, anethole 10.02-13.11%, caryophyllene 2.07-2.88% and borneol acetate 1.3-1.71%.
The density of the traditional Chinese medicine volatile oil extract is 1.0-1.1 g/mL.
The invention also aims to provide the application of the traditional Chinese medicine composition and the traditional Chinese medicine volatile oil extract thereof in preparing antitumor drugs.
The tumor is caused by ras proto-oncogene mutated and then overexpressed.
The tumor is liver cancer, pancreatic cancer, lung cancer and colon cancer.
Since the Ras signaling pathway is highly conserved from nematode to human, in caenorhabditis elegans the let-60 gene encodes a conserved Ras protein, which is 83% homologous to human Ras protein. If ras is mutated, it causes the nematode polynegative phenotype in C.elegans and in humans, malignant proliferation of cells leading to tumorigenesis. Therefore, the model organism Caenorhabditis elegans (Caenorhabditis elegans) Ras/MAPK signal channel overactivation type mutant is widely applied to screening of antitumor drugs for inhibiting Ras channel overactivation as a tumor model.
Therefore, the caenorhabditis elegans mutant is used as a model for evaluating the drugs for inhibiting ras proto-oncogene pathway overactivation, and the conclusion that the traditional Chinese medicine composition and the traditional Chinese medicine volatile oil extract have obvious down-regulation effect on ras proto-oncogene overexpression and have the effect of inhibiting ras proto-oncogene pathway overactivation is obtained, and the traditional Chinese medicine composition and the traditional Chinese medicine volatile oil extract only have the effect on tumor cells caused by ras overactivation caused by let-60 mutation and do not have the effect on tumor cells caused by wild type ras overactivation caused by lin-15 deletion mutation, and the application is safe, namely the caenorhabditis elegans mutant only acts on the cancer cells but does not act on normal cells, can obviously down-regulate ras proto-oncogene overexpression, does not cause gene mutation and does not cause teratogenesis. Therefore, the traditional Chinese medicine composition and the traditional Chinese medicine volatile oil extract can be applied to the preparation of medicines for treating ras proto-oncogene overexpression tumors.
Has the advantages that:
the traditional Chinese medicine composition and the traditional Chinese medicine volatile oil extract can specifically reduce the overexpression caused by ras proto-oncogene mutation, and can be applied to the preparation of medicines for treating malignant tumors caused by the overexpression caused by ras proto-oncogene mutation, particularly the related tumors are liver cancer, pancreatic cancer, lung cancer and colon cancer. The treatment effect of the traditional Chinese medicine composition and the traditional Chinese medicine volatile oil extract on ras gene overexpression tumors does not depend on wide cytotoxic effect, but specifically reduces the overexpression of ras proto-oncogenes; the traditional Chinese medicine composition and the traditional Chinese medicine volatile oil extract only act on tumor cells caused by ras overactivation caused by let-60 mutation, but do not act on tumor cells caused by wild ras overactivation caused by lin-15 deletion mutation, namely the traditional Chinese medicine composition and the traditional Chinese medicine volatile oil extract only act on the cancer cells, do not act on normal cells, do not cause teratogenesis and are safe to apply.
Detailed Description
The following specific examples are provided to illustrate the present invention, but the scope of the claims and the specification of the present invention is not limited to these examples.
EXAMPLE A preparation of an extract of Chinese medicinal volatile oil
The raw material medicaments: flos Caryophylli, fructus Amomi, fructus Anisi Stellati, cortex Cinnamomi, fructus Piperis, radix aucklandiae, and Zingiberis rhizoma
7.5g of clove, 7.5g of amomum villosum, 4.5g of star anise, 12g of cinnamon, 4,5g of white pepper, 4.5g of costustoot and 7.5g of dried ginger.
Pulverizing the above raw materials, adding 10 times of water, and extracting by steam distillation for 3 hr to obtain crude product of Chinese medicinal volatile oil extract.
Extracting the crude product of the Chinese medicinal volatile oil twice by using ether, combining the organic phases obtained by extraction, and volatilizing the ether to obtain the Chinese medicinal volatile oil extract, wherein the obtained Chinese medicinal volatile oil extract is a light yellow oily substance.
And (3) analyzing the components of the volatile oil: GC-MS analysis of the volatile oil extract comprises 42.43-59.19% eugenol, 11.85-15.28% eugenol acetate, 10.02-13.11% anethole, 2.07-2.88% caryophyllene, and 1.3-1.71% borneol acetate.
TABLE 1 GC-MS results of Chinese medicinal volatile oil extracts
EXAMPLE II Effect of extracts of Chinese medicinal volatile oils on caenorhabditis elegans MT2124(let-60 mutation)
Over-activation of Ras in nematodes causes nematode polynegative, in humans, malignant proliferation of cells, and if the test drug inhibits the production of the polynegative phenotype of the test nematode, causing them to develop into wild type, it indicates that the drug acts on the Ras pathway, down-regulates the over-activation of the pathway, and has anti-tumor activity. MT2124 is let-60/ras hyperactivated C.elegans, a multiple vulvar phenotype of nematode species, corresponding to refractory tumors that have mutations in the human ras proto-oncogene. The caenorhabditis elegans MT2124 is treated by the tested medicine, and if the tested medicine can inhibit the generation of the multiple-yin phenotype of the tested caenorhabditis elegans and promote the multi-yin phenotype of the tested caenorhabditis elegans to develop into a wild type, the medicine acts on a Ras pathway to down-regulate the excessive activation of the pathway and has antitumor activity. The higher the proportion of the wild nematode in the tested nematode population is, the better the effect of the tested drug is.
Experimental materials: caenorhabditis elegans MT2124, genotype: let-60(n1046sd, gf) IV, available from CGC; coli OP50, a uracil-leaky mutant, was purchased from CGC as feed for C.elegans.
S, basic liquid: 2.925g of sodium chloride, 0.5g of dipotassium phosphate, 3g of monopotassium phosphate and 500mL of distilled water are shaken to be dissolved and sterilized for 20min at 121 ℃;
1M citrate buffer: 2g of citric acid, 29.35g of potassium citrate and 100mL of distilled water, shaking to dissolve, and sterilizing at 121 ℃ for 20 min;
trace liquid: EDTA0.186g, FeSO4·7H2O 0.06g，MnCl2·4H2O 0.02g，CuSO4·5H2O0.0025g，ZnSO4·7H20.029g of O and 100mL of distilled water, shaking to dissolve, and sterilizing at 121 ℃ for 20 min;
1M magnesium sulfate solution: MgSO (MgSO)4·7H2O24.647 g and distilled water 100mL, shaking to dissolve, sterilizing at 121 deg.C for 20 min;
1M calcium chloride solution: CaCl2·2H2O14.702 g and distilled water 100mL, shaking to dissolve, sterilizing at 121 deg.C for 20 min;
5mg/mL cholesterol solution: 0.05g of cholesterol and 10mL of absolute ethyl alcohol are shaken to be dissolved and stored at 4 ℃ for later use;
s liquid culture medium: 100mL of S base solution, 1mL of 1M citric acid buffer solution, 1mL of Trace solution, 0.3mL of 1M magnesium sulfate solution, 0.3mL of 1M calcium chloride solution and 0.1mL of 5mg/mL cholesterol solution are uniformly mixed.
M9 buffer: 2.486g of sodium chloride, 2.982g of disodium hydrogen phosphate, 1.496g of potassium dihydrogen phosphate, 0.06g of magnesium sulfate and 500mL of distilled water are shaken to dissolve the sodium chloride and the disodium hydrogen phosphate, and the mixture is sterilized at 121 ℃ for 20 min;
the specific experimental steps are as follows:
and (3) culturing nematodes: nematodes were plated on solid NGM plates coated with E.coli OP50 and then incubated in an incubator at 20 ℃ for synchronization when the nematodes grew to adults and eggs and juvenile nematodes were present on the plates.
Nematode synchronization: washing the nematodes on the flat plate down by using an M9 buffer solution, and sucking the washing solution into a centrifugal tube; centrifuging at 4000rpm for 3min, removing supernatant, adding M9 buffer solution, centrifuging, and repeating for several times to clean Escherichia coli; adding lysis solution (final concentration: 3.2% NaClO and 1M NaOH); shaking on vortex instrument for 7min to break nematode to obtain ovum; centrifuging at 4000rpm for 3min, removing supernatant, collecting precipitate to obtain large amount of nematode eggs; the cells were washed twice with M9 buffer, added with 1mL of M9 buffer, and incubated in an incubator at 20 ℃ for 48 hours.
Preparing a liquid medicine: diluting the Chinese medicinal volatile oil extract into the following concentrations:
L：12.5μg/mL；M：25μg/mL；H：50μg/mL；HH：100μg/mL
the medicine has the following functions: the synchronized nematodes are diluted to about 80 per 20. mu.L, and 20. mu.L of the above nematode solution, 120. mu.L of S solution, 10mg/mL of Escherichia coli OP 5020. mu.L, 5mg/mL of cholesterol 20. mu.L and liquid medicine 20. mu.L are added into each well of a 96-well plate, and five replicates are arranged at each concentration. The blank control group was replaced with the same volume of S solution without adding any solution. Culturing at 20 deg.C until the nematode is mature, and detecting the wild type ratio of nematode (namely the ratio of nematode with only one Eumycota) under microscope. The higher proportion of wild type compared to the blank control group indicates a stronger inhibition of ras activation by the drug. The results are shown in the following table:
TABLE 2 influence of Chinese medicinal volatile oil extract on the wild type proportion of caenorhabditis elegans MT2124(let-60 mutation)
Is P < 0.05 compared with blank group
As can be seen from Table 2, the traditional Chinese medicine volatile oil extract enables the wild type proportion of caenorhabditis elegans MT2124 to be obviously increased and to be in a concentration dependence relationship, which shows that the traditional Chinese medicine volatile oil extract acts on a Ras passage and has an obvious down-regulation effect on excessive activation of the Ras/MAPK passage, so that the mutant is changed into the wild type. Therefore, the traditional Chinese medicine volatile oil extract can act on a Ras pathway, down-regulates the over-activation of the pathway, has anti-tumor activity, and can be applied to the preparation of medicines for treating Ras proto-oncogene over-expression tumors.
Example III Effect of Chinese herb volatile oil extract on caenorhabditis elegans MT8189(lin-15 mutation)
The lin-15 is positioned at the upstream of ras and is an inhibitory gene of ras, and after the lin-15 is deleted and mutated, the inhibitory effect on ras is disappeared, and the nematode shows a polynegative phenotype. The MT8189(lin-15) nematode contains wild type let60/ras copies, and when the traditional Chinese medicine volatile oil extract can obviously increase the proportion of the wild type of the caenorhabditis elegans MT2124(let-60) and cannot inhibit the polyvulval phenotype of the MT8189(lin-15) nematode, the traditional Chinese medicine volatile oil extract only plays a role in tumor cells caused by ras over-activation caused by let-60 mutation and does not play a role in tumor cells caused by wild type ras over-activation caused by lin-15 deletion mutation. Therefore, the traditional Chinese medicine volatile oil extract only acts on tumor cells caused by ras over-activation caused by let-60 mutation, but does not act on normal cells.
Experimental materials: caenorhabditis elegans MT8189, multiple vulva phenotype, genotype lin-15(n765ts) X, purchased from CGC; coli OP50, a uracil-leaky mutant, was purchased from CGC as feed for C.elegans.
The specific experimental steps are as follows: the same as the second embodiment.
TABLE 3 Effect of Chinese medicinal volatile oil extract on caenorhabditis elegans MT8189(lin-15 deletion mutation)
As shown in Table 3, the traditional Chinese medicine volatile oil extract of the invention cannot inhibit the multiple-yin phenotype of lin-15(n765), and according to the general principle of genetics, the experimental results of the second example show that the traditional Chinese medicine volatile oil extract of the invention can reverse the multiple-yin phenotype caused by the excessive activation of let60/ras protooncogene at the downstream of the traditional Chinese medicine volatile oil extract, namely, the traditional Chinese medicine volatile oil extract of the invention cannot inhibit the multiple-yin phenotype of let60/ras upstream gene lin-15(n 765). This is because the lin-15(n765) nematode contains 60/ras copies of wild type let, and the volatile oil extract of the traditional Chinese medicine of the invention only acts on the tumor with over-activation of let60/ras caused by let-60 mutation and is insensitive to the tumor with over-activation of let60/ras caused by the deletion mutation of lin-15, i.e. the volatile oil extract of the traditional Chinese medicine of the invention only acts on the tumor cells with over-activation of ras caused by let-60 mutation and does not act on normal cells.
Example four effects of extracts of Chinese medicinal volatile oils on caenorhabditis elegans CB1275(lin-1 deletion mutant)
lin-1 is a gene downstream of ras, is a nuclear transcription factor that can be phosphorylated by MAPK, is a negative regulator of vulvar development, and can regulate the signaling pathway of vulvar development downstream of let-60. lin-1 forms a complex with lin-31, lin-1 encodes a member of the ETS family, and lin-31 encodes a winged helix transcript, and when ras signaling is not activated, lin-1 interacts with lin-31 to form a complex that binds to the initiator portion of the ovipositor function gene and prevents its expression. After the lin-1 deletion mutation, a complex is not formed, the complex is not formed, and the complex cannot be combined with the initiation part of the function gene of the ovipositor, and the gene of the ovipositor is expressed, thereby generating the phenotype of multiple vulva by the development of the false vulva (Ferguson, 1989).
The lin-1 is a gene at the downstream of ras and is a negative regulatory factor for vulva development, after the lin-1 is deleted and mutated, an ovipositor gene is expressed, nematodes show a multiple vulva phenotype, and the Chinese medicinal volatile oil extract cannot inhibit the multiple vulva phenotype of the lin-1, so that the medicament action site is positioned at the upstream of the lin-1 but not the lin-1, namely the medicament down-regulates the over-activation of ras but not directly inhibits the development of vulva cells.
Experimental materials: caenorhabditis elegans genotype: lin-1(e1275) IV was from CGC (Caenorhabditis Genetics center), purchased from CGC; coli OP50, a uracil-leaky mutant, was purchased from CGC as feed for C.elegans.
The specific experimental steps are as follows: same as the second embodiment
The results of example two show that the drug inhibits the nematode polynegative phenotype, but this result only suggests that the drug significantly down-regulates the site of let-60/ras overactivation at or downstream of let-60, a drug capable of inhibiting the polynegative phenotype of MT2124(let-60 mutation), and treatment of c.elegans CB1275 similarly indicates that the drug site of action is upstream of lin-1 if it is not capable of inhibiting its polynegative phenotype. In addition, since lin-1 is a nuclear transcription factor, it interacts with lin-31 to form a complex, regulating the process of development of the vulvar cells. If the medicine has obvious inhibiting effect on the multiple-yin phenotype of the gene inactivation mutation, the medicine can directly inhibit the development of the yin precursor cells and prevent the formation of the multiple-yin phenotype, namely, can inhibit ras proto-oncogene overexpression anti-tumor.
TABLE 4 Effect of Chinese medicinal volatile oil extract on caenorhabditis elegans CB1275(lin-1 deletion mutation)
As shown in Table 4, the multiple-yin phenotype of the CB1275 strain nematode is not influenced by the traditional Chinese medicine volatile oil extract, which indicates that the drug acts on a Ras signal path to inhibit the multiple-yin phenotype formed by Ras over-activation caused by let-60 mutation and cannot inhibit the multiple-yin phenotype formed by lin-1 deletion mutation, and the invention proves that the traditional Chinese medicine volatile oil extract acts on Ras but not acts on lin-1, and the drug down-regulates the Ras over-activation but not directly inhibits the proliferation and differentiation of cells, namely the traditional Chinese medicine volatile oil extract acts on Ras path mechanism dependence rather than extensive cytotoxicity.
Example five effects of extracts of volatile oils from Chinese herbs and individual herbs on caenorhabditis elegans MT2124(let-60 mutation)
Experimental materials: the same as the second embodiment.
The dosage of each single medicine is consistent with that of the Chinese medicinal volatile oil extract.
TABLE 5 Compound crude drug amount of single medicine and Chinese medicinal volatile oil extract
The specific experimental steps are as follows: the same as the second embodiment.
TABLE 6 comparison of the effect of the Chinese medicinal volatile oil extract and the clove volatile oil on caenorhabditis elegans MT2124(let-60 mutation)
TABLE 7 comparison of the effect of the Chinese medicinal volatile oil extract and fructus Amomi volatile oil on caenorhabditis elegans MT2124(let-60 mutation)
TABLE 8 comparison of the effects of Chinese medicinal volatile oil extract and Illicium verum volatile oil on caenorhabditis elegans MT2124(let-60 mutation)
TABLE 9 comparison of the effect of the extract of Chinese medicinal volatile oil and the cinnamon volatile oil on caenorhabditis elegans MT2124(let-60 mutation)
TABLE 10 comparison of the effect of the Chinese medicinal volatile oil extract and the white pepper volatile oil on caenorhabditis elegans MT2124(let-60 mutation)
TABLE 11 comparison of the effect of the Chinese medicinal volatile oil extract and the radix aucklandiae volatile oil on caenorhabditis elegans MT2124(let-60 mutation)
TABLE 12 comparison of the effect of the volatile oil extracts of Chinese herbs and the volatile oil of dried ginger on caenorhabditis elegans MT2124(let-60 mutation)
As shown in tables 6-12, the anti-tumor activity of the volatile oil of the traditional Chinese medicine extract prepared by the preparation method of the invention is obviously higher than that of the volatile oil of a single medicine.
Example five effects of extracts of Chinese medicinal volatile oils and eugenol on caenorhabditis elegans MT2124(let-60 mutation)
The prior art is searched to find that the eugenol has the anti-tumor activity, so that the effect of the traditional Chinese medicine volatile oil extract and the eugenol in inhibiting ras proto-oncogene over-activated tumors is compared in the embodiment, and the experimental results are as follows:
TABLE 13 comparison of the Effect of Chinese medicinal volatile oil extract and eugenol on caenorhabditis elegans MT2124(let-60 mutation)
As shown in table 13, although eugenol also exhibits the effect of inhibiting ras proto-oncogene overexpression, the anti-tumor activity of the traditional Chinese medicine volatile oil extract disclosed by the invention is significantly higher than that of eugenol, so that the traditional Chinese medicine volatile oil extract obtained by reasonable proportioning and extraction treatment realizes the technical effect of synergistic interaction.
Therefore, the invention relates to the application of the traditional Chinese medicine composition and the volatile oil extract thereof in treating ras proto-oncogene overexpression tumors, and particularly relates to the application of the traditional Chinese medicine composition and the volatile oil extract thereof in treating ras proto-oncogene overexpression tumors, such as liver cancer, pancreatic cancer, lung cancer and colon cancer. The treatment effect of the traditional Chinese medicine composition and the traditional Chinese medicine volatile oil extract on ras gene overexpression tumors does not depend on wide cytotoxic effect, but specifically reduces the overexpression of ras proto-oncogenes; the traditional Chinese medicine composition and the traditional Chinese medicine volatile oil extract only act on tumor cells caused by ras over-activation caused by let-60 mutation, but do not act on tumor cells caused by wild ras over-activation caused by lin-15 deletion mutation, namely the traditional Chinese medicine composition and the traditional Chinese medicine volatile oil extract only act on cancer cells, do not act on normal cells, do not cause teratogenesis and are safe to apply.
Claims (4)
1. A traditional Chinese medicine volatile oil extract for resisting colon cancer is characterized by being prepared from 5 parts of clove, 5 parts of fructus amomi, 3 parts of star anise, 8 parts of cinnamon, 3 parts of white pepper, 3 parts of costustoot and 5 parts of dried ginger, and the preparation method comprises the following steps: adding distilled water into the clove, the amomum villosum, the star anise, the cinnamon, the white pepper, the elecampane and the dried ginger according to the proportion, extracting by adopting a steam distillation method to obtain a crude volatile oil extract product, extracting the crude volatile oil extract product by using an organic solvent, volatilizing the solvent from an organic phase obtained by extraction, and dehydrating and drying to obtain the traditional Chinese medicine volatile oil extract.
2. The traditional Chinese medicine volatile oil extract as claimed in claim 1, wherein the marked chemical components of the traditional Chinese medicine volatile oil extract comprise eugenol 42.43-59.19%, eugenol acetate 13.05-15.28%, anethole 10.02-13.11%, caryophyllene 2.07-2.88%, and borneol acetate 1.3-1.71%.
3. The traditional Chinese medicine volatile oil extract as claimed in claim 2, wherein the density of the traditional Chinese medicine volatile oil extract is 1.0-1.1 g/mL.
4. The use of the Chinese medicinal volatile oil extract in preparing an antitumor medicament according to claim 1, wherein the tumor is caused by the overexpression of ras proto-oncogene after mutation, and the tumor is colon cancer.
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