CN102641448A - Anti-lung-cancer traditional Chinese medicine composite, preparation method of composite, and application of composite in preparation of anti-lung-cancer medicament - Google Patents

Anti-lung-cancer traditional Chinese medicine composite, preparation method of composite, and application of composite in preparation of anti-lung-cancer medicament Download PDF

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CN102641448A
CN102641448A CN2012101486840A CN201210148684A CN102641448A CN 102641448 A CN102641448 A CN 102641448A CN 2012101486840 A CN2012101486840 A CN 2012101486840A CN 201210148684 A CN201210148684 A CN 201210148684A CN 102641448 A CN102641448 A CN 102641448A
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semen coicis
bulbus lilii
extraction
ganoderma
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CN102641448B (en
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陈彦
杜萌
曹伟
贺俊杰
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Jiangsu Provincial Insititute of Traditional Chinese Medicine
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Abstract

The invention discloses an anti-lung-cancer traditional Chinese medicine composite, a preparation method of the composite, and an application of the composite in preparation of an anti-lung-cancer medicament. The composite comprises the following active pharmaceutical ingredients in parts by weight: 5 to 40 parts of ganoderma lucidum, 5 to 40 parts of semen coicis and 1 to 10 parts of lily; and as optimization, the composite comprises 20 parts of ganoderma lucidum, 20 parts of semen coicis and 3 parts of lily. The composite consists of six active ingredients including total triterpenoid of the ganoderma lucidum, coix seed oil, total alkaloids of the lily, ganoderma lucidum polysaccharides, semen coicis polysaccharides and lily polysaccharides. The composite disclosed by the invention has an obvious inhibition effect on the growth of Lewis tumors, the antitumor activity of the composite is superior to that of a single component after compatibility administration, and the tumor inhibition rate of the composite can reach 55.13%. Therefore, the active ingredients in the composite are extracted and separated to the greatest extent by virtue of a modern extraction and separation method, and the collaborative antitumor effect of the composite can better come into play by virtue of compatibility combination.

Description

Anti-pulmonary carcinoma Chinese medicine composition, its preparation method and the application in the preparation anti-lung-cancer medicament thereof
Technical field
The present invention relates to technical field of Chinese medicines, be specifically related to a kind of anti-pulmonary carcinoma Chinese medicine composition, this anti-pulmonary carcinoma Chinese medicine composition method for preparing, and the application of this Chinese medicine composition in the preparation anti-lung-cancer medicament.
Background technology
At present, pulmonary carcinoma has become a big killer who threatens human health.According to domestic and foreign literature, the new cases of global pulmonary carcinoma account for 14% of global various tumor new cases up to 205 020, have the characteristics of rate occurred frequently and high mortality.Under the many-side influence of factors such as environmental pollution, smoking, ionizing radiation, occupational exposure and viral infection; The sickness rate of pulmonary carcinoma and mortality rate are in propradation always; Annual speed with 0.5% increases, and along with the increase of smoking in adolescents number, pulmonary carcinoma is rejuvenation day by day.Show that according to the dead survey result of China Ministry of Public Health resident in 2008 pulmonary carcinoma has replaced hepatocarcinoma to become the first malignant tumor cause of the death of China (account for whole malignant tumor dead 22.7%).Therefore, the diagnosis of pulmonary carcinoma and treatment cause enough attention of medical circle.
Pulmonary carcinoma is divided into small cell lung cancer (SCLC, 20%) and nonsmall-cell lung cancer (NSCLC, 80%), and its pathogenic factor is complicated.The treatment of pulmonary carcinoma is main with operation and radiotherapy, chemotherapy both at home and abroad at present, but the easy normal organ to the patient of these methods causes bigger side effect, and prognosis of patients and survival rate are relatively poor.The Chinese medicine that with " eliminating pathogenic factor for supporting vital QI " is characteristic is being brought into play huge effect aspect the treatment pulmonary carcinoma, and its curative effect is not only consolidated, and toxic and side effects is little, can not cause big injury to body, with chemotherapy and with the effect that can play efficacy enhancing and toxicity reducing.Therefore the anti-lung-cancer medicament of from Chinese medicine and natural drug, seeking and develop high-efficiency low-toxicity is the focus and the difficult point of oncology studies.
Through retrieval; 200910266137.0), " a kind of Chinese medicine of treating cancer " (patent No.: 200710166579.9) Chinese medicine composition that has multiple treatment cancer in the existing patent is like " medicine-food dual-purpose composition of a kind of cancer patient of being applicable to and preparation method thereof the " (patent No.:; " a kind of herbal composite of treating liver, stomach, lung, leukemia and preparation method thereof " (patent No.: 201110125724.5); " a kind of Chinese medicine composition of treating cancer and preparation method thereof " (patent No.: 201110165923.9); 201110145442.1), " Chinese medicine medicine for preventing compositions and application thereof " (201110056786.5) etc. " a kind of Chinese medicine composition and application of treating malignant tumor " (patent No.:; Drug kinds used in these compositionss is more, and how to take with decoction, or preparation technology is simpler; Extract to such an extent that directly make oral formulations behind the extractum; The preparation dose is bigger, though certain therapeutic effect is arranged clinically, extraction ratio of effective constituents is lower; Preparation is thicker, is unfavorable for the quality control of preparation.
(200910031997.6 disclose a kind of self-mciro emulsifying nano composition and preparation method thereof of ganodenic acid extract of anti-tumor activity to the antineoplastic related preparations of patent existing report Ganoderma and Semen Coicis; 201110097796.3 disclose a kind of compound ganoderma spore soft capsule and preparation method thereof; 101010237432.6 disclose a kind of Semen Coicis Chinese medicine medicine for preventing and preparation method thereof); These anti-tumor agents have just utilized Ganoderma total triterpenes, the single active component of Semen Coicis oil; And some component that plays assosting effect is abandoned, and according to bibliographical information [Lin Zhibin, Wang Pengyun. the pharmacological research of Ganoderma spore and active component thereof [J]. Peking University's journal (medicine); 2006,38 (5): 541-547; Zhang Yumei; Yang Junshan, Zhao Yangjing, etc. Semen Coicis chemical constituent and pharmacology activity research progress [J]. Chinese Pharmaceutical Journal; 2002; 37 (1): 8-11], ganoderan and Semen Coicis polysaccharide also have antitumor and immunoregulation effect, and therefore above-mentioned preparation can't embody the characteristics of Chinese medicine multicomponent, many target spots wholistic therapy.In addition; 201010187617.0 disclose the preparation of a kind of anticancer pharmaceutical composition and a kind of Chinese medicine composition; Though being used of Chinese medicine composition (concentrated dry powder of the decoction liquor of medical material itself, medical material, the concentrated extract of decoction liquor or decoction liquor) and polysaccharide component paid attention in this invention, the medical material kind that contains in this pharmaceutical composition is more, and the polysaccharide component is not the medical material that derives from the said composition; Complicated component is difficult to carry out quality control.
Summary of the invention
The purpose of this invention is to provide a kind of Chinese medicine composition and preparation method thereof with effect of anti-lung cancer; And this has the application of Chinese medicine composition in the preparation anti-lung-cancer medicament of effect of anti-lung cancer; Through selecting suitable medicine and suitable active component method for distilling, optimize the proportion relation of active component, finally confirm Chinese medicine composition optimization prescription, optimize component compatibility and preparation of compositions method; Make it bring into play antitumor action safely and effectively, be applicable to clinical practice.
First goal of the invention of the present invention is to realize through following technical scheme:
A kind of anti-pulmonary carcinoma Chinese medicine composition is characterized in that, the weight portion of said composition crude drug is formed as follows: 5 ~ 40 parts of Ganodermas, 5 ~ 40 parts of Semen Coiciss, 1 ~ 10 part of Bulbus Lilii.
Wherein crude drug preferably consumption be: 10 ~ 30 parts of Ganodermas, 10 ~ 30 parts of Semen Coiciss, 1 ~ 6 part of Bulbus Lilii.
Wherein the even more ideal consumption of crude drug is: 3 parts of 20 portions of Bulbus Liliies of 20 parts of Semen Coiciss of Ganoderma.
In recent years, receive the concern of research worker in the exploitation of research aspect the anti-pulmonary carcinoma and new drug thereof as Chinese medicine Ganoderma, Semen Coicis, the Bulbus Lilii of " integration of edible and medicinal herbs " always.The present invention then is made up of this three flavors medical material, and the present Research of each medical material of distinguishing the flavor of is following:
Ganoderma is a Polyporaceae fungus Ganoderma lucidum (Leyss. Ex Fr.) Karst. Ganoderma lucidum(Leyss. ex Fr.) Karst. or Ganoderma Ganoderma sinenseZhao. dry sporophore Xu et Zhang).GUIXIN, lung, liver, kidney channel.Has invigorating QI and tranquilization, the effect of relieving cough and asthma.Be used for cough and asthma due to lung deficiency, asthenia is losed heart etc.Modern scientific research shows that Ganoderma has stronger antitumor action, and its antineoplastic active site is mainly Ganoderma triterpenoids and polysaccharide component.
Semen Coicis is the grass Semen Coicis Coix lacryma-jobiL. var. Ma-yuen(Roman.) dry mature kernal of Stapf.Return spleen, stomach, lung meridian.Has promoting diuresis to eliminate damp pathogen, invigorating the spleen to arrest diarrhea, eliminating impediment, evacuation of pus, the effect of detoxicating and resolving stagnation of pathogens.Be used for lung abscess, acute appendicitis, wart, cancerous protuberance etc.Modern study shows that Semen Coicis has the antineoplastic pharmacological action, contains numerous anti-tumor active ingredients in the core, is mainly esters and fatty acid compound and polysaccharide component.
Bulbus Lilii is the liliaceous plant tiger lily Lilium lancifoliumThunb., Bulbus Lilii Lilium browniiF. E. Brown var. ViridulumBaker) or Lilium tenuifolium Lilium pumilumDC. dry meat scale leaf.GUIXIN, lung meridian.Has nourishing YIN and moistening the lung, the effect of clearing away heart-fire for tranquillization.Be used for deficiency of YIN cough caused by dryness, chronic cough hemoptysis etc.Modern study shows that Bulbus Lilii has better antitumor activity, and its antineoplastic main active is biological alkali components and polysaccharide component.
This Chinese medicine composition is clinical anti-pulmonary carcinoma proved recipe, and the Ganoderma invigorating QI and tranquilization is monarch drug, the Semen Coicis invigorating spleen to remove dampness, and the Bulbus Lilii yin nourishing is calmed the nerves, and is all ministerial drug.Full side has boosting qi and nourishing yin, promoting diuresis to eliminate damp pathogen, and the effect of dispersing tumor softening the hard mass is applicable to malignant tumor deficiency of qi and yin, retention of water-damp person.
Further optimization of the present invention can obtain following technical scheme:
Described anti-pulmonary carcinoma Chinese medicine composition is made up of Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids, ganoderan, Semen Coicis polysaccharide and 6 active components of Bulbus Lilii polysaccharide.Wherein, the weight portion of Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids consists of: Ganoderma total triterpenes 1 ~ 20, Semen Coicis oil 1 ~ 35, Bulbus Lilii total alkaloids 1 ~ 10; Ganoderan wherein, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide are to add by its crude drug and Ganoderma total triterpenes, Semen Coicis oil, crude drug amount 1:1 weight ratio that the Bulbus Lilii total alkaloids is identical.
The weight ratio of described Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids, ganoderan, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide is 18-22:33-37:1:33-37:43-47:3-5.
Optimal proportion is that the weight ratio of described Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids, ganoderan, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide is 20:35:1:35:45:4.
Wherein, the Ganoderma total triterpenes extraction ratio is 3.75%, and purity is 61.50%; The Semen Coicis oil extraction ratio is 6.62%, and purity is 74.35%; Bulbus Lilii total alkaloids extraction ratio is 1.25%, and purity is 59.92%; The ganoderan extraction ratio is 6.56%, and purity is 57.70%; The Semen Coicis polysaccharide extract rate is 8.40%, and purity is 64.42%; The Bulbus Lilii polysaccharide extraction ratio is 4.84%, and purity is 62.53%.
More particularly, anti-pulmonary carcinoma Chinese medicine composition of the present invention is meant the compositions that obtains according to following method for preparing:
Step ⑴:, adopt supercritical CO with the Ganoderma sporophore crushing screening 2Extraction extracts Ganoderma total triterpenes class component;
Step ⑵: the decocte with water of Ganoderma coarse powder residue, filtrating concentrates, ethanol precipitation, collecting precipitation, the deposition water dissolves again, removes insoluble impuritiess such as deproteinize, and concentrate drying finally obtains ganoderan class component;
Step ⑶: the Semen Coicis pulverizing is sieved, adopt supercritical CO 2Extraction extracts Semen Coicis oil;
Step ⑷: the decocte with water of Semen Coicis coarse powder residue, filtrating concentrates, ethanol precipitation, collecting precipitation, the deposition water dissolves again, removes insoluble impuritiess such as deproteinize, and concentrate drying finally obtains Semen Coicis polysaccharide component;
Step ⑸: with the Bulbus Lilii crushing screening, extract with acid ethanol solution, reclaim ethanol, reclaim liquid and alkalize with strong aqua ammonia, the reuse chloroform extraction reclaims chloroform, and drying promptly gets Bulbus Lilii total alkaloids class component;
Step ⑹: the decocte with water of Bulbus Lilii coarse powder residue, filtrating concentrates, ethanol precipitation, collecting precipitation, the deposition water dissolves again, removes insoluble impuritiess such as deproteinize, and concentrate drying finally obtains Bulbus Lilii polysaccharide class component;
Step ⑺:, obtain described anti-pulmonary carcinoma Chinese medicine composition with the component mix homogeneously that above each step obtains.
The technical scheme of accomplishing second invention task of the present invention is: the method for preparing of above-mentioned anti-pulmonary carcinoma Chinese medicine composition: it is characterized in that step is following,
Step ⑴:, adopt supercritical CO with the Ganoderma sporophore crushing screening 2Extraction extracts Ganoderma total triterpenes class component;
Step ⑵: the decocte with water of Ganoderma coarse powder residue, filtrating concentrates, ethanol precipitation, collecting precipitation, the deposition water dissolves again, removes insoluble impuritiess such as deproteinize, and concentrate drying finally obtains ganoderan class component;
Step ⑶: the Semen Coicis pulverizing is sieved, adopt supercritical CO 2Extraction extracts Semen Coicis oil;
Step ⑷: the decocte with water of Semen Coicis coarse powder residue, filtrating concentrates, ethanol precipitation, collecting precipitation, the deposition water dissolves again, removes insoluble impuritiess such as deproteinize, and concentrate drying finally obtains Semen Coicis polysaccharide component;
Step ⑸: with the Bulbus Lilii crushing screening, extract with acid ethanol solution, reclaim ethanol, reclaim liquid and alkalize with strong aqua ammonia, the reuse chloroform extraction reclaims chloroform, and drying promptly gets Bulbus Lilii total alkaloids class component;
Step ⑹: the decocte with water of Bulbus Lilii coarse powder residue, filtrating concentrates, ethanol precipitation, collecting precipitation, the deposition water dissolves again, removes insoluble impuritiess such as deproteinize, and concentrate drying finally obtains Bulbus Lilii polysaccharide class component;
Step ⑺:, obtain described anti-pulmonary carcinoma Chinese medicine composition with the component mix homogeneously that above each step obtains.
The further optimization of above method for preparing, the concrete operations step is:
Step ⑴:, adopt supercritical CO with the Ganoderma sporophore crushing screening 2Extraction extracts Ganoderma total triterpenes class component, and extracting pressure is 20 ~ 40MPa, and extraction temperature is 40 ~ 50 ℃, and the extraction time is 1 ~ 3 h, and entrainer (percent by volume 95% ethanol) consumption is 1 ~ 4 mLg -1Crude drug.
Step ⑵: Ganoderma coarse powder residue, the decocting that adds 8 ~ 14 times of amounts boils, and decocting number of times is 1 ~ 3 time; Each 1 ~ 3h, filtrating concentrates, and it is 60 ~ 90% that concentrated solution alcohol is sink to concentration of alcohol (percent by volume); Collecting precipitation, deposition is dissolved with the water of 4 ~ 10 times of amounts again, removes insoluble impuritiess such as deproteinize; Concentrate drying finally obtains ganoderan class component.
Step ⑶: the Semen Coicis pulverizing is sieved, adopt supercritical CO 2Extraction extracts Semen Coicis oil, and extracting pressure is 15 ~ 25MPa, and extraction temperature is 30 ~ 40 ℃, and the extraction time is 1 ~ 3 h, and entrainer (percent by volume 95% ethanol) consumption is 1 ~ 4 mLg -1Crude drug.
Step ⑷: Semen Coicis coarse powder residue, the decocting that adds 8 ~ 14 times of amounts boils, and decocting number of times is 1 ~ 3 time; Each 1 ~ 3 h, filtrating concentrates, and it is 60 ~ 90% that concentrated solution alcohol is sink to concentration of alcohol (percent by volume); Collecting precipitation, deposition is dissolved with the water of 4 ~ 10 times of amounts again, removes insoluble impuritiess such as deproteinize; Concentrate drying finally obtains Semen Coicis polysaccharide component.
Step ⑸: with the Bulbus Lilii crushing screening, the volume fraction that adds 5 ~ 10 times of amounts is that the alcoholic solution (containing 1% acetic acid) of 70%-90% extracts, and extraction time is 1 ~ 3 time; Each 1 ~ 3 h reclaims ethanol, reclaims liquid and alkalizes to pH8 ~ 11 with strong aqua ammonia; Reuse equal amounts of chloroform extraction 2 ~ 8 times; Reclaim chloroform, drying promptly gets Bulbus Lilii total alkaloids class component.
Step ⑹: Bulbus Lilii coarse powder residue, the decocting that adds 8 ~ 14 times of amounts boils, and decocting number of times is 1 ~ 3 time; Each 1 ~ 3 h, filtrating concentrates, and it is 60 ~ 90% that concentrated solution alcohol is sink to concentration of alcohol (percent by volume); Collecting precipitation, deposition is dissolved with the water of 4 ~ 10 times of amounts again, removes insoluble impuritiess such as deproteinize; Concentrate drying finally obtains Bulbus Lilii polysaccharide class component.
Step ⑺:, obtain described anti-pulmonary carcinoma Chinese medicine composition with the component mix homogeneously that above each step obtains.
In the prioritization scheme of Chinese medicine composition method for preparing of the present invention,
The described extraction conditions of step ⑴ is: extracting pressure 25 ~ 35MPa, and extraction temperature is 43 ~ 48 ℃; The extraction time is 1 ~ 2 h, and entrainer (95% ethanol) consumption is 2 ~ 3 mLg -1Crude drug;
The described decocte with water of step ⑵ is the water that adds 9 ~ 11 times of amounts, and decocting number of times is 1 ~ 2 time, each 1 ~ 2h;
Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 75 ~ 85% to concentrated solution alcohol;
Described deposition is that the water with 5 ~ 7 times of amounts dissolves again;
The described extraction conditions of step ⑶ is: extracting pressure is 20 ~ 25MPa, and extraction temperature is 35 ~ 40 ℃, and the extraction time is 2 ~ 3 h, and entrainer (95% ethanol) consumption is 2 ~ 3 mLg -1Crude drug;
The described decocte with water of step ⑷ is the water that adds 9 ~ 11 times of amounts, and decocting number of times is 1 ~ 2 time, each 1 ~ 2 h;
Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 75 ~ 85% to concentrated solution alcohol;
Described deposition is that the water with 5 ~ 7 times of amounts dissolves again;
The described ethanol acetate that adds of step ⑸ extracts, and the volume fraction that is 6 ~ 8 times of amounts of adding is 75% ~ 85% alcoholic solution (containing percent by volume 1% acetic acid) extraction, and extraction time is 1 ~ 2 time, each 1 ~ 2 h;
Described filtrating is to alkalize to pH9 ~ 10 with strong aqua ammonia;
The extraction of reuse equal amounts of chloroform is 3 ~ 7 times;
The described decocte with water of step ⑹ is the water that adds 9 ~ 11 times of amounts, and decocting number of times is 1 ~ 2 time, each 1 ~ 2 h;
Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 75 ~ 85% to concentrated solution alcohol;
Described deposition is that the water with 5 ~ 7 times of amounts dissolves again.
In the prioritization scheme of Chinese medicine composition method for preparing, the condition that the present invention recommends is:
The described extraction conditions of step ⑴ is: extracting pressure is 30 MPa, and extraction temperature is 45 ℃; The extraction time is 1.5 h, and entrainer (percent by volume 95% ethanol) consumption is 3 mLg -1Crude drug;
The described decocte with water of step ⑵ is the water that adds 10 times of amounts, and decocting number of times is 2 times, each 1 h;
Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 80% to concentrated solution alcohol;
Described deposition is that the water with 6 times of amounts dissolves again;
The described extraction conditions of step ⑶ is: extracting pressure is 22MPa, and extraction temperature is 38 ℃; The extraction time is 2.5 h, and entrainer (95% ethanol) consumption is 3 mLg -1Crude drug;
The described decocte with water of step ⑷ is the water that adds 10 times of amounts, and decocting number of times is 2 times, each 1 h;
Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 80% to concentrated solution alcohol;
Described deposition is that the water with 6 times of amounts dissolves again;
The described ethanol acetate that adds of step ⑸ extracts, and the volume fraction that is 7 times of amounts of adding is 80% alcoholic solution (containing 1% acetic acid) extraction, and extraction time is 2 times, each 1 h;
Described filtrating is to alkalize to pH10 with strong aqua ammonia;
The extraction of reuse equal amounts of chloroform is 5 times;
The described decocte with water of step ⑹ is the water that adds 10 times of amounts, and decocting number of times is 2 times, each 1 h;
Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 80% to concentrated solution alcohol;
Described deposition is that the water with 6 times of amounts dissolves again.
The technical scheme of accomplishing the 3rd invention task of the present invention is: the application of above-mentioned anti-pulmonary carcinoma Chinese medicine composition in the preparation anti-lung-cancer medicament.
The optimization prescription of anti-pulmonary carcinoma Chinese medicine composition provided by the invention, its prescription and compositions proportioning screening process are following:
1. on cellular level, detect the inhibitory action of the active component of different proportionings to growth of tumour cell.
Cell strain: people's pulmonary carcinoma A549 and SPC-A-1 cell strain.
Because the polysaccharide component is at external acellular inhibition proliferation function, so this experiment selects for use the proportion relation of Ganoderma total triterpenes, Semen Coicis oil and Bulbus Lilii total alkaloids to come the optimum ratio prescription.
The weight portion of known Ganoderma, Semen Coicis and Bulbus Lilii clinical prescription consumption consists of: Ganoderma 10, Semen Coicis 10, Bulbus Lilii 6, and the extraction ratio of active component is respectively Ganoderma total triterpenes 3.75%, Semen Coicis oil 6.62% and Bulbus Lilii total alkaloids 1.25% in the three flavor medical materials.Therefore; With Ganoderma total triterpenes, Semen Coicis oil, 3 kinds of active components of Bulbus Lilii total alkaloids is the influence factor; On original clinical basis with the prescription amount, utilization equivalent multiplication method (0.5 times, 1 times, 2 times) is in conjunction with medical material, active component content; Three dosage confirming each active component are the level that influences, and adopt L 9(3 4) the orthogonal test table design that experimentizes.Be made into different effective component crowd according to the different dosages proportioning; Selecting two kinds of human lung carcinoma cell lines of A549 and SPC-A-1 for use is model; Through the inhibitory action of each active component crowd on cell proliferation of MTT activity test detection, investigate its anti-tumor activity size, with half-inhibition concentration IC 50Be index, the best proportioning dosage of final preferred active component.Orthogonal test factor level harmony in the exterior L 9(3 4) orthogonal experiments sees table 1,2.
 
Table 1 orthogonal test factor level table
Figure 956099DEST_PATH_IMAGE001
Table 2 L 9(3 4) orthogonal experiments
Figure 257767DEST_PATH_IMAGE002
Carry out intuitive analysis and variance analysis for ease of the pair of orthogonal result of the test; So introduce a balance index " degree of membership "; To the IC50 standardization; Index degree of membership=(index maximum-desired value)/(index maximum-index minima), maximum, minima that wherein index is maximum, minima is respectively the IC50 that surveys, desired value be the desired value that will calculate.The analysis result of A549 cell is seen table 3, and 5 see table 4,6 with the analysis result of SPC-A-1 cell.
 
Table 3 L 9(3 4) orthogonal experiments
The factor experiment A B C D IC 50Degree of membership
1 1 1 1 1 0.30
2 1 2 2 2 0.25
3 1 3 3 3 0.00
4 2 1 2 3 0.26
5 2 2 3 1 0.01
6 2 3 1 2 1.00
7 3 1 3 2 0.23
8 3 2 1 3 0.98
9 3 3 2 1 0.75
k1 0.55 0.79 2.28 1.06 ?
k2 1.27 1.24 1.26 1.48 ?
k3 1.96 1.75 0.24 1.24 ?
k1/3 0.18 0.26 0.76 0.35 ?
k2/3 0.42 0.41 0.42 0.49 ?
k3/3 0.65 0.58 0.08 0.41 ?
R 0.47 0.32 0.68 0.14 ?
Table 4 L 9(3 4) orthogonal experiment data result table
The factor experiment A B C D IC 50Degree of membership
1 1 1 1 1 0.23
2 1 2 2 2 0.31
3 1 3 3 3 0.00
4 2 1 2 3 0.27
5 2 2 3 1 0.11
6 2 3 1 2 1.00
7 3 1 3 2 0.24
8 3 2 1 3 0.95
9 3 3 2 1 0.84
k1 0.54 0.74 2.18 1.18 ?
k2 1.38 1.37 1.42 1.55 ?
k3 2.03 1.84 0.35 1.22 ?
k1/3 0.18 0.25 0.73 0.39 ?
k2/3 0.46 0.46 0.47 0.52 ?
k3/3 0.68 0.61 0.12 0.41 ?
R 0.50 0.37 0.61 0.12 ?
Table 5 analysis of variance table
Soruces of variation Sum of deviation square Degree of freedom Mean square F Significance
A 0.33 2.000 0.166 11.196 0.082
B 0.15 2.000 0.077 5.196 0.161
C 0.69 2.000 0.347 23.432 0.041
Error 0.03 2.000 0.015 ? ?
Annotate: F (0.05 2,2)=19.00
Table 6 analysis of variance table
Soruces of variation Sum of deviation square Degree of freedom Mean square F Significance
A 0.37 2.000 0.186 13.534 0.069
B 0.20 2.000 0.102 7.388 0.119
C 0.56 2.000 0.282 20.499 0.047
Error 0.03 2.000 0.014 ? ?
Annotate: F (0.05 2,2)=19.00
Comprehensive A549 and SPC-A-1 cell experiment result can find out according to directly perceived and variance analysis, to half-inhibition concentration IC 50The factor that has the greatest impact is C (a Bulbus Lilii total alkaloids), secondly is the A factor, and B influences minimum, and optimum combination is A 3B 3C 1, promptly Bulbus Lilii total alkaloids size of shared weight portion in compositions has a strong impact on the anti-tumor activity size of said composition, and its proportion is big more, its IC 50Big more.Therefore; The best proportioning of Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids is 750 mg in this antitumor Chinese: 1324 mg: 37.5 mg, promptly the best compatibility consumption of crude drug is (weight ratio): Ganoderma 20 weight portions, Semen Coicis 20 weight portions, Bulbus Lilii 3 weight portions.
In vivo on the animal model antitumor action to each active component and compatibility component thereof carry out evaluating drug effect.
It is 1 * 10 that cell density is transferred to 7The Lewis cell suspension inoculation of individual/mL is subcutaneous in the right armpit of C57BL/6 mice, every 0.2 mL.Next day after the inoculation, weigh random packet, 8 every group.Administering mode: gastric infusion.Blank group: normal saline, every day 0.4 mL/ time; Positive controls: cyclophosphamide (20 mgg -1D -1), every day 0.2 mL/ time; Drug group: Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids, Ganoderma total triterpenes+Semen Coicis oil (in the combination of crude drug amount 1:1 ratio), Ganoderma total triterpenes+Semen Coicis oil+Bulbus Lilii total alkaloids (in the combination of crude drug amount 20:20:3 ratio), Ganoderma total triterpenes+Semen Coicis oil+Bulbus Lilii total alkaloids+ganoderan, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide (make up in crude drug amount 20:20:3 ratio; Ganoderan wherein, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide are to add by its crude drug and Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids crude drug crude drug amount 1:1 weight ratio), all establish high low dose group (high dose: the total crude drug kg of 30 g -1D -1, low dosage: the total crude drug kg of 15 g -1D -1), every day 0.4 mL/ time, continuous 14 d.Drug withdrawal next day, weigh, pull out eyeball and get blood, blood sample is in centrifugal 5 min of 5000 rpm, and-20 ℃ of freezing preservations are subsequent use, take off neck and put to death mice, peel off oxter tumor, liver, thymus and spleen.Calculate tumour inhibiting rate, liver index, thymus index and spleen index, the content of TNF-α and IL-6 in the mensuration mice plasma.The result sees table 7,8,9.
Tumour inhibiting rate (%)=(the average tumor of the average tumor weight-administration of model group group is heavy)/average tumor of model group heavy * 100%
Liver index=liver weight (mg)/mice body weight (g)
Thymus index=thymic weight (mg)/mice body weight (g)
Spleen index=spleen weight (mg)/mice body weight (g)
Table 7 active component and compatibility component thereof to the inhibitory action of Lewis tumor-bearing mice tumor (
Figure 618342DEST_PATH_IMAGE003
± S, n=8)
Annotate: * is each group and the comparison of normal saline group (* p < 0.05)
The result shows with the normal saline group and compares; Except Bulbus Lilii total alkaloids low dose group, resulting Ganoderma total triterpenes of the present invention and the high low dose group of Semen Coicis oil, Bulbus Lilii total alkaloids high dose group and each active component compositions all have inhibitory action to Lewis growth of tumor in the body.After Ganoderma total triterpenes, Semen Coicis oil and the administration of Bulbus Lilii total alkaloids compatibility; Its anti-tumor activity is superior to one-component (p < 0.05); And after Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids and ganoderan, Semen Coicis polysaccharide, the administration of Bulbus Lilii polysaccharide compositions; Tumour inhibiting rate is the highest, reaches 55.13%, and the tumour inhibiting rate of joining stilbene PINGXIAO JIA0NANG (suppression ratio to transplantability Lewis lung cancer tumor is 42.18%) than the Chinese medicine of present national approved is high; Explain and adopt modern extraction separation means, can bring into play the synergistic antitumor effect better the active component in this pharmaceutical composition farthest extraction separation and compatibility.
Table 8 active component and compatibility component thereof to the influence of Lewis tumor-bearing mice immune organ (
Figure 437579DEST_PATH_IMAGE003
± S, n=8)
Figure 390491DEST_PATH_IMAGE005
Annotate: * is each group and the comparison of normal saline group (* p < 0.05); # is each group and cyclophosphamide group relatively (# p < 0.05)
The result shows, compares with the normal saline group, and liver index, spleen index, the thymus index of cyclophosphamide administration group mice all have certain reduction, possibly have inhibitory action to cause to the mouse immune system owing to cyclophosphamide.Compare the animal immune index of Ganoderma total triterpenes high dose group, Semen Coicis oil high dose group and each compositions high dose group of active component be improved (p < 0.05) with cyclophosphamide administration group.After Ganoderma total triterpenes, Semen Coicis oil, the administration of Bulbus Lilii total alkaloids compatibility, compare with the one-component administration, liver index, spleen index and the thymus index of mice all has significance to increase (p < 0.05).After the Ganoderma total triterpenes that the present invention obtains, Semen Coicis oil, Bulbus Lilii total alkaloids and the administration of polysaccharide compositions; Compare with the cyclophosphamide positive controls; Liver index, spleen index and the thymus index of mice all has significance to increase (P < 0.05); Explain that the compositions that the present invention obtains can be active through enhancing human body immunity, and the performance antitumor action.
Table 9 active component and compatibility component thereof to TNF-a in the Lewis tumor-bearing mice blood plasma with
The comparison of IL-6 content (
Figure 187546DEST_PATH_IMAGE003
± S, n=8)
Annotate: * is each group and the comparison of normal saline group (* p < 0.05); # is each group and cyclophosphamide group relatively (# p < 0.05)
The result shows, compares with the normal saline group, and the TNF-a and the IL-6 content of cyclophosphamide administration group significantly reduce, and possibly have inhibitory action to cause to the mouse immune system owing to cyclophosphamide; Compare with the cyclophosphamide positive controls, after the Ganoderma total triterpenes that the present invention obtains, Semen Coicis oil, Bulbus Lilii total alkaloids and the administration of polysaccharide compositions, TNF-a in the mice plasma and the content of IL-6 all have significance to increase (P < 0.05).Explain that the compositions that the present invention obtains all has regulating action to tumor-bearing mice cells in vivo vitamin T NF-a and IL-6, can strengthen antitumor action through the activity of the human body immunity improving factor.
Chinese medicine composition of the present invention extracts purification effectively with fat-soluble active component in the medical material and polysaccharide component, and the extraction method for preparing that adopts is fit to, and industry is big produces.Through the reasonable compatibility between the component; With the anti-pulmonary carcinoma experiment in inside and outside is foundation; Final best prescription and the best proportioning dosage of compositions of confirming that anti-tumor activity is the strongest; Make the anti-lung cancer activity of the Chinese medicine composition that the present invention obtains apparently higher than one-component, demonstrated fully the treatment advantage and the characteristic of the anti-pulmonary carcinoma of Chinese medicine compound " multicomponent, multipath, many target spots ".
The specific embodiment
Embodiment 1, anti-pulmonary carcinoma Chinese medicine composition, and the weight portion of crude drug is formed as follows: 20 parts of step ⑴ Ganodermas, 20 parts of Semen Coiciss, 3 parts of Bulbus Liliies.With the Ganoderma sporophore crushing screening, adopt supercritical CO 2Extraction extracts Ganoderma total triterpenes class component, and extracting pressure is 30 MPa, and extraction temperature is 45 ℃; The extraction time is 1.5 h, and entrainer (95% ethanol) consumption is 3 mLg -1Crude drug.Step ⑵: Ganoderma coarse powder residue, the decocting that adds 10 times of amounts boils, and decocting number of times is 2 times, each 1 h; Filtrating concentrates, and it is 80% that concentrated solution alcohol is sink to concentration of alcohol (percent by volume); Collecting precipitation, deposition is dissolved with the water of 6 times of amounts again; Remove insoluble impuritiess such as deproteinize, concentrate drying finally obtains ganoderan class component.Step ⑶: the Semen Coicis pulverizing is sieved, adopt supercritical CO 2Extraction extracts Semen Coicis oil, and extracting pressure is 22MPa, and extraction temperature is 38 ℃; The extraction time is 2.5 h, and entrainer (95% ethanol) consumption is 3 mLg -1Crude drug; Step ⑷: Semen Coicis coarse powder residue, the decocting that adds 10 times of amounts boils, and decocting number of times is 2 times, each 1 h; Filtrating concentrates, and it is 80% that concentrated solution alcohol is sink to concentration of alcohol (percent by volume); Collecting precipitation, deposition is dissolved with the water of 6 times of amounts again; Remove insoluble impuritiess such as deproteinize, concentrate drying finally obtains Semen Coicis polysaccharide component.Step ⑸: with the Bulbus Lilii crushing screening, the volume fraction that adds 7 times of amounts is 80% alcoholic solution (containing 1% acetic acid) extraction, and extraction time is 2 times; Each 1 h reclaims ethanol, reclaims liquid and alkalizes to pH10 with strong aqua ammonia; Reuse equal amounts of chloroform extraction 5 times; Reclaim chloroform, drying promptly gets Bulbus Lilii total alkaloids class component.Step ⑹: Bulbus Lilii coarse powder residue, the decocting that adds 10 times of amounts boils, and decocting number of times is 2 times, each 1 h; Filtrating concentrates, and it is 80% that concentrated solution alcohol is sink to concentration of alcohol (percent by volume); Collecting precipitation, deposition is dissolved with the water of 6 times of amounts again, removes insoluble impuritiess such as deproteinize, and concentrate drying finally obtains Bulbus Lilii polysaccharide class component.Step ⑺:, obtain described anti-pulmonary carcinoma Chinese medicine composition with the component mix homogeneously that above each step obtains.Said composition is used to treat pulmonary carcinoma.It is 1 * 10 that cell density is transferred to 7The Lewis cell suspension inoculation of individual/mL is subcutaneous in the right armpit of C57BL/6 mice, every 0.2 mL.Next day after the inoculation, weigh random packet, 8 every group.Administering mode: gastric infusion.Blank group: normal saline, every day 0.4 mL/ time; Positive controls: cyclophosphamide (20 mgg -1D -1), every day 0.2 mL/ time; Drug group: the Chinese medicine composition of the Ganoderma total triterpenes that the present invention obtains, Semen Coicis oil, Bulbus Lilii total alkaloids, ganoderan, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide, dosage: high dose 30 g crude drug kg -1D -1, low dosage 15 g crude drug kg -1D -1, every day 0.4 mL/ time, continuous 14 d.Drug withdrawal next day, weigh, pull out eyeball and get blood, blood sample is in centrifugal 5 min of 5000 rpm, and-20 ℃ of freezing preservations are subsequent use, take off neck and put to death mice, peel off oxter tumor, liver, thymus and spleen.Calculate tumour inhibiting rate, liver index, thymus index and spleen index, the content of TNF-α and IL-6 in the mensuration mice plasma.Tumour inhibiting rate (%)=(the average tumor of the average tumor weight-administration of model group group is heavy)/average tumor of model group heavy * 100%; Liver index=liver weight (mg)/mice body weight (g); Thymus index=thymic weight (mg)/mice body weight (g); Spleen index=spleen weight (mg)/mice body weight (g).
Experimental result shows that the tumor weight of this Chinese medicine composition high and low dose group is starkly lower than the normal saline group, and tumour inhibiting rate is respectively 55.13%, 40.77%, compares with other administration groups, and tumour inhibiting rate has significance to improve (P < 0.05).Compare with the normal saline group, liver index, thymus index and the spleen index of this Chinese medicine composition high and low dose group are improved; With positive controls relatively, liver index, thymus index and the spleen index of this Chinese medicine composition high and low dose group all has significance to increase (P < 0.05).With normal saline group and cyclophosphamide group relatively, the TNF-a in the mice plasma of this Chinese medicine composition high and low dose group and the content of IL-6 all have significance increase (P < 0.05).
Embodiment 2, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 5 parts of Ganodermas, 5 parts of Semen Coiciss, 1 part of Bulbus Lilii.Simultaneously, the described extraction conditions of step ⑴ is: pressure 25MPa, and extraction temperature is 43 ℃; The extraction time is 1 h, and entrainer (95% ethanol) consumption is 1 mLg -1Crude drug; The described decocte with water of step ⑵ is that the decocting that adds 8 times of amounts boils, and decocting number of times is 1 time, each 2 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 75% to concentrated solution alcohol; Described deposition is that the water with 4 times of amounts dissolves again; The described extraction conditions of step ⑶ is: extracting pressure is 20MPa, and extraction temperature is 35 ℃, and the extraction time is 2 h, and entrainer (95% ethanol) consumption is 1 mLg -1Crude drug; The described decocte with water of step ⑷ is that the decocting that adds 8 times of amounts boils, and decocting number of times is 1 time, each 1 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 75% to concentrated solution alcohol; Described deposition is that the water with 4 times of amounts dissolves again; The described ethanol acetate that adds of step ⑸ extracts, and the volume fraction that is 8 times of amounts of adding is 70% alcoholic solution (containing 1% acetic acid) extraction, and extraction time is 1 time, each 2 h; Described recovery liquid is to alkalize to pH9 with strong aqua ammonia; The extraction of reuse equal amounts of chloroform is 3 times; The described decocte with water of step ⑹ is that the decocting that adds 8 times of amounts boils, and decocting number of times is 1 time, each 1 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 75% to concentrated solution alcohol; Described deposition is that the water of 4 times of amounts dissolves again.
Embodiment 3, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 40 parts of Ganodermas, 40 parts of Semen Coiciss, 10 parts of Bulbus Liliies.Simultaneously, the described extraction conditions of step ⑴ is: pressure 35MPa, and extraction temperature is 48 ℃; The extraction time is 2 h, and entrainer (95% ethanol) consumption is 2 mLg -1Crude drug; The described decocte with water of step ⑵ is that the decocting that adds 13 times of amounts boils, and decocting number of times is 2 times, each 2 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 85% to concentrated solution alcohol; Described deposition is that the water with 10 times of amounts dissolves again; The described extraction conditions of step ⑶ is: extracting pressure is 25MPa, and extraction temperature is 40 ℃, and the extraction time is 3 h, and entrainer (95% ethanol) consumption is 2 mLg -1Crude drug; The described decocte with water of step ⑷ is that the decocting that adds 13 times of amounts boils, and decocting number of times is 2 times, each 2 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 85% to concentrated solution alcohol; Described deposition is that the water with 10 times of amounts dissolves again; The described ethanol acetate that adds of step ⑸ extracts, and the volume fraction that is 9 times of amounts of adding is 85% alcoholic solution (containing 1% acetic acid) extraction, and extraction time is 2 times, each 2 h; Described recovery liquid is to alkalize to pH10 with strong aqua ammonia; The extraction of reuse equal amounts of chloroform is 7 times; The described decocte with water of step ⑹ is that the decocting that adds 13 times of amounts boils, and decocting number of times is 2 times, each 2 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 85% to concentrated solution alcohol; Described deposition is that the water of 10 times of amounts dissolves again.
Embodiment 4, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 5 parts of Ganodermas, 40 parts of Semen Coiciss, 1 part of Bulbus Lilii.Simultaneously, the described extraction conditions of step ⑴ is: pressure 35MPa, and extraction temperature is 43 ℃; The extraction time is 2 h, and entrainer (95% ethanol) consumption is 2 mLg -1Crude drug; The described decocte with water of step ⑵ is that the decocting that adds 11 times of amounts boils, and decocting number of times is 2 times, each 1 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 85% to concentrated solution alcohol; Described deposition is that the water with 5 times of amounts dissolves again; The described extraction conditions of step ⑶ is: extracting pressure is 25MPa, and extraction temperature is 35 ℃, and the extraction time is 3 h, and entrainer (95% ethanol) consumption is 2 mLg -1Crude drug; The described decocte with water of step ⑷ is that the decocting that adds 11 times of amounts boils, and decocting number of times is 2 times, each 1 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 85% to concentrated solution alcohol; Described deposition is that the water with 5 times of amounts dissolves again; The described ethanol acetate that adds of step ⑸ extracts, and the volume fraction that is 8 times of amounts of adding is 85% alcoholic solution (containing 1% acetic acid) extraction, and extraction time is 2 times, each 1 h; Described recovery liquid is to alkalize to pH9 with strong aqua ammonia; The extraction of reuse equal amounts of chloroform is 7 times; The described decocte with water of step ⑹ is that the decocting that adds 11 times of amounts boils, and decocting number of times is 2 times, each 1 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 85% to concentrated solution alcohol; Described deposition is that the water of 5 times of amounts dissolves again.
Embodiment 5, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 40 parts of Ganodermas, 5 parts of Semen Coiciss, 10 parts of Bulbus Liliies.Simultaneously, the described extraction conditions of step ⑴ is: pressure 25MPa, and extraction temperature is 48 ℃; The extraction time is 1 h, and entrainer (95% ethanol) consumption is 1 mLg -1Crude drug; The described decocte with water of step ⑵ is that the decocting that adds 13 times of amounts boils, and decocting number of times is 1 time, each 2 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 75% to concentrated solution alcohol; Described deposition is that the water with 7 times of amounts dissolves again; The described extraction conditions of step ⑶ is: extracting pressure is 20MPa, and extraction temperature is 40 ℃, and the extraction time is 2 h, and entrainer (95% ethanol) consumption is 1 mLg -1Crude drug; The described decocte with water of step ⑷ is that the decocting that adds 13 times of amounts boils, and decocting number of times is 1 time, each 2 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 75% to concentrated solution alcohol; Described deposition is that the water with 7 times of amounts dissolves again; The described ethanol acetate that adds of step ⑸ extracts, and the volume fraction that is 6 times of amounts of adding is 75% alcoholic solution (containing 1% acetic acid) extraction, and extraction time is 1 time, each 2 h; Described recovery liquid is to alkalize to pH10 with strong aqua ammonia; The extraction of reuse equal amounts of chloroform is 3 times; The described decocte with water of step ⑹ is that the decocting that adds 13 times of amounts boils, and decocting number of times is 1 time, each 2 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 75% to concentrated solution alcohol; Described deposition is that the water of 7 times of amounts dissolves again.
Embodiment 6, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 10 parts of Ganodermas, 10 parts of Semen Coiciss, 1 part of Bulbus Lilii.The described extraction conditions of step ⑴ is: pressure 20MPa, and extraction temperature is 40 ℃; The extraction time is 1 h, and entrainer (95% ethanol) consumption is 1 mLg -1Crude drug; The described decocte with water of step ⑵ is that the decocting that adds 8 times of amounts boils, and decocting number of times is 1 time, each 1 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 60% to concentrated solution alcohol; Described deposition is that the water with 4 times of amounts dissolves again; The described extraction conditions of step ⑶ is: extracting pressure is 15MPa, and extraction temperature is 30 ℃, and the extraction time is 1 h, and entrainer (95% ethanol) consumption is 1 mLg -1Crude drug; The described decocte with water of step ⑷ is that the decocting that adds 8 times of amounts boils, and decocting number of times is 1 time, each 1 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 60% to concentrated solution alcohol; Described deposition is that the water with 4 times of amounts dissolves again; The described ethanol acetate that adds of step ⑸ extracts, and the volume fraction that is 5 times of amounts of adding is 70% alcoholic solution (containing 1% acetic acid) extraction, and extraction time is 1 time, each 1 h; Described recovery liquid is to alkalize to pH8 with strong aqua ammonia; The extraction of reuse equal amounts of chloroform is 2 times; The described decocte with water of step ⑹ is that the decocting that adds 8 times of amounts boils, and decocting number of times is 1 time, each 1 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 60% to concentrated solution alcohol; Described deposition is that the water of 4 times of amounts dissolves again.
Embodiment 7, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 30 parts of Ganodermas, 30 parts of Semen Coiciss, 6 parts of Bulbus Liliies.The described extraction conditions of step ⑴ is: pressure 40MPa, and extraction temperature is 50 ℃; The extraction time is 3 h, and entrainer (95% ethanol) consumption is 4 mLg -1Crude drug; The described decocte with water of step ⑵ is that the decocting that adds 14 times of amounts boils, and decocting number of times is 3 times, each 3 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 90% to concentrated solution alcohol; Described deposition is that the water with 10 times of amounts dissolves again; The described extraction conditions of step ⑶ is: extracting pressure is 25MPa, and extraction temperature is 40 ℃, and the extraction time is 3 h, and entrainer (95% ethanol) consumption is 4 mLg -1Crude drug; The described decocte with water of step ⑷ is that the decocting that adds 14 times of amounts boils, and decocting number of times is 3 times, each 3 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 90% to concentrated solution alcohol; Described deposition is that the water with 10 times of amounts dissolves again; The described ethanol acetate that adds of step ⑸ extracts, and the volume fraction that is 10 times of amounts of adding is 90% alcoholic solution (containing 1% acetic acid) extraction, and extraction time is 3 times, each 3 h; Described recovery liquid is to alkalize to pH11 with strong aqua ammonia; The extraction of reuse equal amounts of chloroform is 8 times; The described decocte with water of step ⑹ is that the decocting that adds 14 times of amounts boils, and decocting number of times is 3 times, each 3 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 90% to concentrated solution alcohol; Described deposition is that the water of 10 times of amounts dissolves again.
Embodiment 8, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 10 parts of Ganodermas, 30 parts of Semen Coiciss, 1 part of Bulbus Lilii.The described extraction conditions of step ⑴ is: pressure 40MPa, and extraction temperature is 40 ℃; The extraction time is 3 h, and entrainer (95% ethanol) consumption is 3 mLg -1Crude drug; The described decocte with water of step ⑵ is that the decocting that adds 9 times of amounts boils, and decocting number of times is 3 times, each 1 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 90% to concentrated solution alcohol; Described deposition is that the water with 6 times of amounts dissolves again; The described extraction conditions of step ⑶ is: extracting pressure is 25MPa, and extraction temperature is 30 ℃, and the extraction time is 3 h, and entrainer (95% ethanol) consumption is 3 mLg -1Crude drug; The described decocte with water of step ⑷ is that the decocting that adds 9 times of amounts boils, and decocting number of times is 3 times, each 1 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 90% to concentrated solution alcohol; Described deposition is that the water with 6 times of amounts dissolves again; The described ethanol acetate that adds of step ⑸ extracts, and the volume fraction that is 7 times of amounts of adding is 90% alcoholic solution (containing 1% acetic acid) extraction, and extraction time is 3 times, each 1 h; Described recovery liquid is to alkalize to pH8 with strong aqua ammonia; The extraction of reuse equal amounts of chloroform is 8 times; The described decocte with water of step ⑹ is that the decocting that adds 9 times of amounts boils, and decocting number of times is 3 times, each 1 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 90% to concentrated solution alcohol; Described deposition is that the water of 6 times of amounts dissolves again.
Embodiment 9, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 30 parts of Ganodermas, 10 parts of Semen Coiciss, 6 parts of Bulbus Liliies.The described extraction conditions of step ⑴ is: pressure 20MPa, and extraction temperature is 50 ℃; The extraction time is 1 h, and entrainer (95% ethanol) consumption is 2 mLg -1Crude drug; The described decocte with water of step ⑵ is that the decocting that adds 14 times of amounts boils, and decocting number of times is 1 time, each 3 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 60% to concentrated solution alcohol; Described deposition is that the water with 9 times of amounts dissolves again; The described extraction conditions of step ⑶ is: extracting pressure is 15MPa, and extraction temperature is 40 ℃, and the extraction time is 1 h, and entrainer (95% ethanol) consumption is 2 mLg -1Crude drug; The described decocte with water of step ⑷ is that the decocting that adds 14 times of amounts boils, and decocting number of times is 1 time, each 3 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 60% to concentrated solution alcohol; Described deposition is that the water with 9 times of amounts dissolves again; The described ethanol acetate that adds of step ⑸ extracts, and the volume fraction that is 10 times of amounts of adding is 90% alcoholic solution (containing 1% acetic acid) extraction, and extraction time is 1 time, each 3 h; Described recovery liquid is to alkalize to pH11 with strong aqua ammonia; The extraction of reuse equal amounts of chloroform is 2 times; The described decocte with water of step ⑹ is that the decocting that adds 14 times of amounts boils, and decocting number of times is 1 time, each 3 h; Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 60% to concentrated solution alcohol; Described deposition is that the water of 9 times of amounts dissolves again.
Embodiment 10, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 5 parts of Ganodermas, 5 parts of Semen Coiciss, 10 parts of Bulbus Liliies.
Embodiment 11, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 5 parts of Ganodermas, 40 parts of Semen Coiciss, 10 parts of Bulbus Liliies.
Embodiment 12, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 40 parts of Ganodermas, 5 parts of Semen Coiciss, 1 part of Bulbus Lilii.
Embodiment 13, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 40 parts of Ganodermas, 40 parts of Semen Coiciss, 1 part of Bulbus Lilii.
Embodiment 14, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 30 parts of Ganodermas, 30 parts of Semen Coiciss, 1 part of Bulbus Lilii.
Embodiment 15, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 10 parts of Ganodermas, 30 parts of Semen Coiciss, 6 parts of Bulbus Liliies.
Embodiment 16, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 10 parts of Ganodermas, 10 parts of Semen Coiciss, 6 parts of Bulbus Liliies.
Embodiment 17, and is basic identical with embodiment 1, but following change is arranged, and the weight portion of crude drug is formed as follows: 30 parts of Ganodermas, 10 parts of Semen Coiciss, 1 part of Bulbus Lilii.
Embodiment 18, and is basic identical with embodiment 1, but following change is arranged, and wherein, the weight portion of Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids consists of: Ganoderma total triterpenes 1, Semen Coicis oil 1, Bulbus Lilii total alkaloids 1; Ganoderan wherein, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide are to add by its crude drug and Ganoderma total triterpenes, Semen Coicis oil, crude drug amount 1:1 weight ratio that the Bulbus Lilii total alkaloids is identical.
Embodiment 19, and is basic identical with embodiment 1, but following change is arranged, and wherein, the weight portion of Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids consists of: Ganoderma total triterpenes 20, Semen Coicis oil 35, Bulbus Lilii total alkaloids 10; Ganoderan wherein, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide are to add by its crude drug and Ganoderma total triterpenes, Semen Coicis oil, crude drug amount 1:1 weight ratio that the Bulbus Lilii total alkaloids is identical.
Embodiment 20, and is basic identical with embodiment 1, but following change is arranged, and wherein, the weight portion of Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids consists of: Ganoderma total triterpenes 1, Semen Coicis oil 35, Bulbus Lilii total alkaloids 1; Ganoderan wherein, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide are to add by its crude drug and Ganoderma total triterpenes, Semen Coicis oil, crude drug amount 1:1 weight ratio that the Bulbus Lilii total alkaloids is identical.
Embodiment 21, and is basic identical with embodiment 1, but following change is arranged, and wherein, the weight portion of Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids consists of: Ganoderma total triterpenes 1, Semen Coicis oil 1, Bulbus Lilii total alkaloids 10; Ganoderan wherein, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide are to add by its crude drug and Ganoderma total triterpenes, Semen Coicis oil, crude drug amount 1:1 weight ratio that the Bulbus Lilii total alkaloids is identical.
Embodiment 22, and is basic identical with embodiment 1, but following change is arranged, and wherein, the weight portion of Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids consists of: Ganoderma total triterpenes 20, Semen Coicis oil 1, Bulbus Lilii total alkaloids 1; Ganoderan wherein, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide are to add by its crude drug and Ganoderma total triterpenes, Semen Coicis oil, crude drug amount 1:1 weight ratio that the Bulbus Lilii total alkaloids is identical.
Embodiment 23, and is basic identical with embodiment 1, but following change is arranged, and wherein, the weight portion of Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids consists of: Ganoderma total triterpenes 20, Semen Coicis oil 35, Bulbus Lilii total alkaloids 1; Ganoderan wherein, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide are to add by its crude drug and Ganoderma total triterpenes, Semen Coicis oil, crude drug amount 1:1 weight ratio that the Bulbus Lilii total alkaloids is identical.
Embodiment 24, and is basic identical with embodiment 1, but following change is arranged, and wherein, the weight portion of Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids consists of: Ganoderma total triterpenes 20, Semen Coicis oil 1, Bulbus Lilii total alkaloids 10; Ganoderan wherein, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide are to add by its crude drug and Ganoderma total triterpenes, Semen Coicis oil, crude drug amount 1:1 weight ratio that the Bulbus Lilii total alkaloids is identical.
Embodiment 25, and is basic identical with embodiment 1, but following change is arranged, and wherein, the weight portion of Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids consists of: Ganoderma total triterpenes 1, Semen Coicis oil 35, Bulbus Lilii total alkaloids 10; Ganoderan wherein, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide are to add by its crude drug and Ganoderma total triterpenes, Semen Coicis oil, crude drug amount 1:1 weight ratio that the Bulbus Lilii total alkaloids is identical.
Embodiment 26, and is basic identical with embodiment 1, but following change is arranged, and the weight ratio of described Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids, ganoderan, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide is 20:35:1:35:45:4.
Embodiment 27, and is basic identical with embodiment 1, but following change is arranged, and the weight ratio of described Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids, ganoderan, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide is 18:33:1:33:43:3.
Embodiment 28, and is basic identical with embodiment 1, but following change is arranged, and the weight ratio of described Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids, ganoderan, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide is 22:37:1:37:47:5.
Embodiment 29, and is basic identical with embodiment 1, but following change is arranged, and the weight ratio of described Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids, ganoderan, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide is 18:37:1:33:47:3.
Embodiment 30, and is basic identical with embodiment 1, but following change is arranged, and the weight ratio of described Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids, ganoderan, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide is 22:33:1:37:43:5.
Embodiment 31, and is basic identical with embodiment 1, but following change is arranged, in the described compositions wherein the Ganoderma total triterpenes extraction ratio be 3.75%, purity is 61.50%; The Semen Coicis oil extraction ratio is 6.62%, and purity is 74.35%; Bulbus Lilii total alkaloids extraction ratio is 1.25%, and purity is 59.92%; The ganoderan extraction ratio is 6.56%, and purity is 57.70%; The Semen Coicis polysaccharide extract rate is 8.40%, and purity is 64.42%; The Bulbus Lilii polysaccharide extraction ratio is 4.84%, and purity is 62.53%.

Claims (10)

1. an anti-pulmonary carcinoma Chinese medicine composition is characterized in that, the weight portion of said composition crude drug is formed as follows: Ganoderma 5 ~ 40, Semen Coicis 5 ~ 40, Bulbus Lilii 1 ~ 10.
2. anti-pulmonary carcinoma Chinese medicine composition according to claim 1 is characterized in that the weight portion of said crude drug consists of: Ganoderma 10 ~ 30, Semen Coicis 10 ~ 30, Bulbus Lilii 1 ~ 6.
3. anti-pulmonary carcinoma Chinese medicine composition according to claim 2 is characterized in that the weight portion of said crude drug consists of: Ganoderma 20, Semen Coicis 20, Bulbus Lilii 3.
4. according to the described anti-pulmonary carcinoma Chinese medicine composition of one of claim 1-3; It is characterized in that; Said composition is made up of Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids, ganoderan, Semen Coicis polysaccharide and 6 active components of Bulbus Lilii polysaccharide; Wherein, the weight portion of Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids consists of: Ganoderma total triterpenes 1 ~ 20, Semen Coicis oil 1 ~ 35, Bulbus Lilii total alkaloids 1 ~ 10; Ganoderan wherein, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide add by its crude drug and Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids crude drug crude drug amount 1:1 weight ratio.
5. anti-pulmonary carcinoma Chinese medicine composition according to claim 4; It is characterized in that the weight ratio of described Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids, ganoderan, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide is 18-22:33-37:1:33-37:43-47:3-5.
6. anti-pulmonary carcinoma Chinese medicine composition according to claim 5 is characterized in that the weight ratio of described Ganoderma total triterpenes, Semen Coicis oil, Bulbus Lilii total alkaloids, ganoderan, Semen Coicis polysaccharide and Bulbus Lilii polysaccharide is 20:35:1:35:45:4; Wherein, the Ganoderma total triterpenes extraction ratio is 3.75%, and purity is 61.50%; The Semen Coicis oil extraction ratio is 6.62%, and purity is 74.35%; Bulbus Lilii total alkaloids extraction ratio is 1.25%, and purity is 59.92%; The ganoderan extraction ratio is 6.56%, and purity is 57.70%; The Semen Coicis polysaccharide extract rate is 8.40%, and purity is 64.42%; The Bulbus Lilii polysaccharide extraction ratio is 4.84%, and purity is 62.53%.
7. the method for preparing of the said anti-pulmonary carcinoma Chinese medicine composition of claim 1 is characterized in that step is following:
Step ⑴:, adopt supercritical CO with the Ganoderma sporophore crushing screening 2Extraction extracts Ganoderma total triterpenes class component;
Step ⑵: the decocte with water of Ganoderma coarse powder residue, filtrating concentrates, ethanol precipitation, collecting precipitation, the deposition water dissolves again, removes insoluble impuritiess such as deproteinize, and concentrate drying finally obtains ganoderan class component;
Step ⑶: the Semen Coicis pulverizing is sieved, adopt supercritical CO 2Extraction extracts Semen Coicis oil;
Step ⑷: the decocte with water of Semen Coicis coarse powder residue, filtrating concentrates, ethanol precipitation, collecting precipitation, the deposition water dissolves again, removes insoluble impuritiess such as deproteinize, and concentrate drying finally obtains Semen Coicis polysaccharide component;
Step ⑸: with the Bulbus Lilii crushing screening, extract with acid ethanol solution, reclaim ethanol, reclaim liquid and alkalize with strong aqua ammonia, the reuse chloroform extraction reclaims chloroform, and drying promptly gets Bulbus Lilii total alkaloids class component;
Step ⑹: the decocte with water of Bulbus Lilii coarse powder residue, filtrating concentrates, ethanol precipitation, collecting precipitation, the deposition water dissolves again, removes insoluble impuritiess such as deproteinize, and concentrate drying finally obtains Bulbus Lilii polysaccharide class component;
Step ⑺:, obtain described anti-pulmonary carcinoma Chinese medicine composition with the component mix homogeneously that above each step obtains.
8. according to the method for preparing of the said anti-pulmonary carcinoma Chinese medicine composition of claim 7, it is characterized in that the concrete operations step is:
Step ⑴:, adopt supercritical CO with the Ganoderma sporophore crushing screening 2Extraction extracts Ganoderma total triterpenes class component, and extracting pressure is 20 ~ 40MPa, and extraction temperature is 40 ~ 50 ℃; The extraction time is 1 ~ 3 h, and entrainer (95% ethanol) consumption is 1 ~ 4 mLg -1Crude drug;
Step ⑵: Ganoderma coarse powder residue, the decocting that adds 8 ~ 14 times of amounts boils, and decocting number of times is 1 ~ 3 time; Each 1 ~ 3h, filtrating concentrates, and it is 60 ~ 90% that concentrated solution alcohol is sink to concentration of alcohol (percent by volume); Collecting precipitation, deposition is dissolved with the water of 4 ~ 10 times of amounts again, removes insoluble impuritiess such as deproteinize; Concentrate drying finally obtains ganoderan class component;
Step ⑶: the Semen Coicis pulverizing is sieved, adopt supercritical CO 2Extraction extracts Semen Coicis oil, and extracting pressure is 15 ~ 25MPa, and extraction temperature is 30 ~ 40 ℃, and the extraction time is 1 ~ 3 h, and entrainer (95% ethanol) consumption is 1 ~ 4 mLg -1Crude drug;
Step ⑷: Semen Coicis coarse powder residue, the decocting that adds 8 ~ 14 times of amounts boils, and decocting number of times is 1 ~ 3 time; Each 1 ~ 3 h, filtrating concentrates, and it is 60 ~ 90% that concentrated solution alcohol is sink to concentration of alcohol (percent by volume); Collecting precipitation, deposition is dissolved with the water of 4 ~ 10 times of amounts again, removes insoluble impuritiess such as deproteinize; Concentrate drying finally obtains Semen Coicis polysaccharide component;
Step ⑸: with the Bulbus Lilii crushing screening, the volume fraction that adds 5 ~ 10 times of amounts is that the alcoholic solution (containing 1% acetic acid) of 70%-90% extracts, and extraction time is 1 ~ 3 time; Each 1 ~ 3 h reclaims ethanol, reclaims liquid and alkalizes to pH8 ~ 11 with strong aqua ammonia; Reuse equal amounts of chloroform extraction 2 ~ 8 times; Reclaim chloroform, drying promptly gets Bulbus Lilii total alkaloids class component;
Step ⑹: Bulbus Lilii coarse powder residue, the decocting that adds 8 ~ 14 times of amounts boils, and decocting number of times is 1 ~ 3 time; Each 1 ~ 3 h, filtrating concentrates, and it is 60 ~ 90% that concentrated solution alcohol is sink to concentration of alcohol (percent by volume); Collecting precipitation, deposition is dissolved with the water of 4 ~ 10 times of amounts again, removes insoluble impuritiess such as deproteinize; Concentrate drying finally obtains Bulbus Lilii polysaccharide class component;
Step ⑺:, obtain described anti-pulmonary carcinoma Chinese medicine composition with the component mix homogeneously that above each step obtains.
9. the method for preparing of said according to Claim 8 anti-pulmonary carcinoma Chinese medicine composition is characterized in that,
The described extraction conditions of step ⑴ is: extracting pressure is 30 MPa, and extraction temperature is 45 ℃; The extraction time is 1.5 h, and entrainer (95% ethanol) consumption is 3 mLg -1Crude drug;
The described decocte with water of step ⑵ is that the decocting that adds 10 times of amounts boils, and decocting number of times is 2 times, each 1 h;
Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 80% to concentrated solution alcohol;
Described deposition is that the water with 6 times of amounts dissolves again;
The described extraction conditions of step ⑶ is: extracting pressure is 22MPa, and extraction temperature is 38 ℃; The extraction time is 2.5 h, and entrainer (95% ethanol) consumption is 3 mLg -1Crude drug;
The described decocte with water of step ⑷ is that the decocting that adds 10 times of amounts boils, and decocting number of times is 2 times, each 1 h;
Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 80% to concentrated solution alcohol;
Described deposition is that the water with 6 times of amounts dissolves again;
The described ethanol acetate that adds of step ⑸ extracts, and the volume fraction that is 7 times of amounts of adding is 80% alcoholic solution (containing 1% acetic acid) extraction, and extraction time is 2 times, each 1 h;
Described recovery liquid is to alkalize to pH10 with strong aqua ammonia;
The extraction of reuse equal amounts of chloroform is 5 times;
The described decocte with water of step ⑹ is that the decocting that adds 10 times of amounts boils, and decocting number of times is 2 times, each 1 h;
Described concentrated solution precipitate with ethanol condition is that to be sink to concentration of alcohol (percent by volume) be 80% to concentrated solution alcohol;
Described deposition is that the water with 6 times of amounts dissolves again.
10. the application of the described anti-pulmonary carcinoma Chinese medicine composition of claim 1 in the preparation anti-lung-cancer medicament.
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CN102940813A (en) * 2012-11-16 2013-02-27 陈彦 Self-microemulsifying nanocombination comprising lucid ganoderma total triterpene and jobstears seed oil and preparation method and application thereof
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