A traditional chinese medicine composition for metastatic non-small cell lung cancer

Abstract

The application discloses a traditional Chinese medicine composition for treating metastatic non-small cell lung cancer, which comprises the following medicinal materials in parts by weight: Lysimachia clethophylla 1-20 parts, Lysimachia christinae 1-20 parts, chrysanthemum 1-15 parts, Atractylodes lancea 1-15 parts, Atractylodes chinensis 1-10 parts and Amomum villosum 1-10 parts. The application further provides a medicine prepared from the traditional Chinese medicine composition and a preparation method of the medicine, and application of the traditional Chinese medicine composition and the medicine prepared from the traditional Chinese medicine composition in preparation of medicines for treating non-small cell lung cancer, especially metastatic non-small cell lung cancer.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition and application thereof in treating metastatic non-small cell lung cancer. BACKGROUND

[0002] Lung cancer is one of the most important cancer types in new cancer cases and cancer-induced deaths each year. About 85% of all new lung cancer cases are non-small cell lung cancer (NSCLC). Due to the lack of effective early screening methods, most NSCLC patients show local or distant metastasis at the time of initial diagnosis; at this time, the patients have missed the opportunity for surgical cure. Non-surgical drug treatment is an important treatment method for patients with metastatic non-small cell lung cancer who cannot undergo surgery and after surgery. Drugs for metastatic non-small cell lung cancer have always been a hot spot in medical research.

[0003] Lung cancer belongs to the category of "Xire", "Lung Accumulation", "Lung Atrophy", "Cough", "Hemoptysis", "Chest Pain", etc. in traditional Chinese medicine. Most doctors believe that the most important and basic pathogenesis is accumulation of deficiency and damage, which is mainly caused by deficiency of vital qi, exposure to evil toxin, emotional depression, diet damage, and old illness, etc. factors, resulting in dysfunction of viscera, abnormal circulation of qi and blood, pathological changes such as qi stagnation, blood stasis, phlegm turbidity, dampness turbidity, and heat toxin, etc. The accumulation in the lung, mutual knotting, gradually accumulated over time, and finally developed into a kind of malignant disease with deficiency in the root and excess in the branch.

[0004] In view of the characteristics of deficiency in the root and excess in the branch, and complicated deficiency and excess of metastatic non-small cell lung cancer patients, traditional Chinese medicine treatment of metastatic non-small cell lung cancer mainly adopts the methods of clearing heat and dampness, purging fire and detoxifying, eliminating accumulation and resolving stasis, soothing liver and regulating qi, and promoting blood circulation to remove blood stasis, etc. combined with tonifying spleen and qi, nourishing blood and soothing liver, and tonifying yin fluid to support healthy qi and eliminate pathogenic factors. For example, the Chinese invention patent application with publication number CN 114668814A (publication date June 28, 2022) discloses a traditional Chinese medicine compound composition for inhibiting lung cancer metastasis, which is prepared from the following weight parts of traditional Chinese medicine raw materials: Prunella vulgaris 9-40 parts, Huperzia serrata 10-40 parts, Berberis dictyota 10-40 parts, raw oyster 10-40 parts, Rubia cordifolia 10-40 parts, wild grapevine 10-40 parts, Herba Dispori 10-40 parts, and Herba Siphonostegia 10-40 parts. The traditional Chinese medicine compound composition takes Huperzia serrata and Prunella vulgaris as the monarch drug, Herba Siphonostegia, Herba Dispori, and oyster as the ministerial drug, and Berberis dictyota, wild grapevine, and Rubia cordifolia as the auxiliary drug, which has the functions of softening and resolving stasis, clearing heat and detoxifying.

[0005] For example, CN111228439A (publication date June 5, 2020) also discloses a traditional Chinese medicine composition for treating lung cancer and anti-lung cancer metastasis, which is prepared from the following weight parts of traditional Chinese medicine raw materials: American ginseng 15-20 parts, ophiopogon 15-25 parts, schisandra 15-20 parts, peony 15-25 parts, forsythia 15-25 parts, gualou 15-25 parts, Sichuan beimu 15-25 parts, mulberry 15-25 parts, Gastrodia 15-25 parts, dodder 20-30 parts, ginseng root 15-25 parts, platycodon 15-25 parts, chicken gold 15-30 parts, bergamot 15-25 parts, Rhizoma Arisaematis 15-20 parts, orange peel 15-25 parts, dandelion 20-30 parts, peach kernel 15-20 parts, Sichuan lovage 15-25 parts, curcuma 15-25 parts, hainan 25-35 parts, white muscardine 15-25 parts, silkworm chrysalis 15-25 parts, polygonatum 15-25 parts, stone see 20-30 parts, bletilla 15-25 parts, cat's paw 15-25 parts, xianhecao 20-30 parts, madder 15-25 parts, raw turtle shell 25-40 parts, raw turtle shell 25-40 parts, raw licorice 15-20 parts. The traditional Chinese medicine composition is based on the principles of clearing dryness, moistening the lungs, nourishing yin and tonifying the lungs, removing swelling and blood stasis, clearing heat and reducing phlegm, and tonifying the spleen and kidney.

[0006] Although drug researchers and clinical workers are committed to the research and development of non-small cell lung cancer treatment drugs, according to the 2023 global cancer report released by the International Agency for Research on Cancer of the World Health Organization, lung cancer is the first cause of death. Data shows that about 350 people die of lung cancer every day, which is almost 2.5 times the number of deaths from the second leading cause of death (colorectal cancer). Therefore, the clinical treatment demand for lung cancer has not been met. SUMMARY

[0007] In view of the deficiencies of the prior art, the present application provides a traditional Chinese medicine composition for treating non-small cell lung cancer, especially metastatic non-small cell lung cancer. The traditional Chinese medicine composition has a novel composition idea and simple composition; preliminary experiments show that it has a significant inhibitory effect on the growth and metastasis of human non-small cell lung cancer cell lines (NCI-H460 and A549) transplanted tumors, suggesting that it can be used to treat metastatic non-small cell lung cancer.

[0008] In order to achieve the above technical effects, the present application adopts the following technical solutions:

[0009] A traditional Chinese medicine composition for metastatic non-small cell lung cancer, the raw materials include the following weight parts of medicinal materials:

[0010] Lobelia 1-20 parts, money grass 1-20 parts, chrysanthemum 1-15 parts, atractylodes 1-15 parts, atractylodes 1-10 parts, grass fruit kernel 1-10 parts.

[0011] Preferably, the Chinese medicine composition for treating metastatic non-small cell lung cancer comprises the following ingredients by weight:

[0012] Lobelia chinensis Lour. 5-20 parts, Lysimachia christinae Hance 5-20 parts, Chrysanthemum morifolium Ramat. 5-15 parts, Atractylodes macrocephala Koidz. 5-15 parts, Atractylodes lancea DC. 5-10 parts, Amomum kravanh Pierre ex Gagnep. 5-10 parts.

[0013] More preferably, the Chinese medicine composition for treating metastatic non-small cell lung cancer comprises the following ingredients by weight:

[0014] Lobelia chinensis Lour. 10-20 parts, Lysimachia christinae Hance 10-20 parts, Chrysanthemum morifolium Ramat. 5-15 parts, Atractylodes macrocephala Koidz. 5-15 parts, Atractylodes lancea DC. 5-10 parts, Amomum kravanh Pierre ex Gagnep. 5-10 parts.

[0015] As a particularly preferred embodiment, the present application provides a Chinese medicine composition for treating metastatic non-small cell lung cancer, which comprises the following ingredients by weight:

[0016] Lobelia chinensis Lour. 15 parts, Lysimachia christinae Hance 15 parts, Chrysanthemum morifolium Ramat. 10 parts, Atractylodes macrocephala Koidz. 10 parts, Atractylodes lancea DC. 9 parts, Amomum kravanh Pierre ex Gagnep. 6 parts.

[0017] As a preferred embodiment, the present application provides a Chinese medicine composition for treating metastatic non-small cell lung cancer, which is composed of Lobelia chinensis Lour., Atractylodes lancea DC., Amomum kravanh Pierre ex Gagnep., Atractylodes macrocephala Koidz., Lysimachia christinae Hance and Chrysanthemum morifolium Ramat., and the weight parts of each ingredient are as defined above.

[0018] Lobelia chinensis Lour. is the dried whole plant of Lobelia chinensis Lour. of the family Campanulaceae, which is pungent in nature and flat in taste. It belongs to the heart, small intestine and lung channels. It has the effects of diuresis, swelling, heat and detoxification. It is used for abdominal dropsy, edema, carbuncle, snake bite and advanced schistosomiasis.

[0019] Lysimachia christinae Hance is the dried whole plant of Lysimachia christinae Hance of the family Primulaceae, which belongs to the liver, gallbladder, kidney and bladder channels. It is sweet and salty in taste and cold in nature. It has the effects of clearing heat and dampness, removing stones and swelling. It is commonly used for hot and sand stones, urine pain, yellow urine, carbuncle, snake bite, liver and gallbladder stones, and urinary stones.

[0020] Chrysanthemum morifolium Ramat. is the dried flower head of Chrysanthemum morifolium Ramat. of the family Asteraceae, which belongs to the lung and liver channels. It is sweet, bitter and cold in nature. It has the effects of dispelling wind, clearing heat, clearing eyesight and detoxification. It is used for headache, dizziness, red eyes, chest heat, carbuncle and swelling.

[0021] Atractylodes Macrocephala Koidz. of the family Compositae, belonging to the spleen and stomach channels. It is bitter, sweet, and warm. It has the effects of invigorating the spleen, benefiting qi, drying dampness, benefiting water, stopping sweating, and calming the fetus. It is used for spleen deficiency, poor appetite, abdominal distension and diarrhea, phlegm and dizziness, edema, and restless fetus.

[0022] Atractylodes lancea (Thunb.) DC. or Atractylodes chinensis (DC.) Koidz. of the family Compositae, belonging to the spleen, stomach, and liver channels. It is bitter, warm, and dry. It has the effects of drying dampness, invigorating the spleen, dispelling wind and cold, and clearing the eyes. It is used for abdominal distension, diarrhea, edema, beriberi, rheumatism, colds, and night blindness.

[0023] Amomum tsao-ko Crevost et Lemaire of the Zingiberaceae family, which is a traditional medicine and food plant. It is warm and bitter, and belongs to the spleen and stomach channels. It has the effects of drying dampness, warming the middle, and eliminating malaria and phlegm. It is used for malaria, phlegm, abdominal pain, vomiting, diarrhea, and food accumulation.

[0024] Among the above six medicinal materials, lobelia and atractylodes are the monarch drug. The Medical Record of Western Medicine in Chinese Medicine describes atractylodes as follows: "It is helpful to supplement the lung when used with cooling and moistening drugs; it is good for regulating the liver when used with ascending and dispersing drugs; it is also good for nourishing the heart when used with tranquilizing drugs; and it is helpful for kidney when used with yin-nourishing drugs. It has the fullness of earth and is the essential drug of acquired vital energy, so it can be used for all organs of metal, wood, water, fire, and viscera, which can benefit from it." Therefore, lobelia and atractylodes have the effects of invigorating the spleen, clearing heat, and resolving toxins. Money grass and chrysanthemum are the ministerial drugs, which enhance the effects of clearing heat, benefiting dampness, and resolving toxins. Both lobelia and money grass are slightly cold and sweet, which are beneficial to dampness without harming yin. Atractylodes and grassfruit are used together to help dry dampness, warm the middle, and resolve phlegm, which are the auxiliary drugs. The combination of the above drugs balances cold and heat, clears and supplements, and attacks and supplements. The whole prescription has the effects of clearing heat and resolving toxins, invigorating the spleen, and eliminating dampness.

[0025] The present application also aims to provide a medicine, which includes the above-mentioned traditional Chinese medicine composition and can also include or not include pharmaceutically acceptable excipients.

[0026] The pharmaceutically acceptable adjuvant of the present application includes, but is not limited to, (1) diluents, such as starch, powdered sugar, dextrin, lactose, pregelatinized starch, microcrystalline fiber, inorganic calcium salt (such as calcium sulfate, calcium hydrogen phosphate, medicinal calcium carbonate, etc.), mannitol, vegetable oil, polyethylene glycol, etc.; (2) binders, such as distilled water, ethanol, starch paste, sodium carboxymethyl cellulose, hydroxypropyl cellulose, methyl cellulose and ethyl cellulose, hydroxypropyl methyl cellulose, etc.; (3) disintegrants, such as dry starch, sodium carboxymethyl starch, low-substitution hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, etc.; (4) lubricants, such as magnesium stearate, microcrystalline silica, talc, hydrogenated vegetable oil, polyethylene glycol, magnesium lauryl sulfate, etc.; (5) solvents, such as water, ethanol solution, etc.

[0027] The drug is any one of the clinically acceptable preparations, preferably oral preparations, selected from the group consisting of decoction, powder, capsule, tablet, honeyed pill, water-honeyed pill, water pill, concentrated pill, paste pill, wax pill, granule, oral liquid or drop pill, more preferably decoction or granule.

[0028] The third object of the present application is to provide a preparation method of the above-mentioned drug, comprising preparing each medicinal material according to the weight parts, and preparing a clinically acceptable preparation by adding or not adding pharmaceutically acceptable adjuvant according to the conventional method in the art.

[0029] In addition, the present application also provides the use of the above-mentioned traditional Chinese medicine composition, the above-mentioned drug or the drug prepared according to the above-mentioned preparation method in the preparation of a drug for treating non-small cell lung cancer, especially metastatic non-small cell lung cancer.

[0030] The traditional Chinese medicine composition and the drug prepared therefrom can be used for the treatment of metastatic non-small cell lung cancer which cannot be resected by surgery, the treatment of metastatic non-small cell lung cancer after surgery, and the treatment of metastatic non-small cell lung cancer which relapses after surgery.

[0031] The traditional Chinese medicine composition for treating metastatic non-small cell lung cancer of the present application has a novel prescription, although only six ingredients, but it has a significant inhibitory effect on the growth and metastasis of non-small cell lung cancer cell lines (NCI-H460 and A549) transplanted tumors, therefore, it is suggested that the traditional Chinese medicine composition of the present application can be used for treating metastatic non-small cell lung cancer.

[0032] In the present specification, the "weight parts" of each component represents the amount ratio relationship between each component, rather than the actual mass unit. According to the actual situation, the unit weight part can be any mass, such as 1 g, 500 g or 1 kg, even 15 g, 30 g, etc. DETAILED DESCRIPTION

[0033] The present application will be described below with reference to specific examples. Those skilled in the art will understand that the examples are only for illustration of the present application and do not limit the scope of the present application in any way.

[0034] The experimental methods in the following examples are all conventional methods unless otherwise specified. The medicinal material raw materials, reagent materials and the like used in the following examples are all commercially purchased products unless otherwise specified.

[0035] Example 1 A traditional Chinese medicine composition for metastatic non-small cell lung cancer

[0036] The raw material composition of the traditional Chinese medicine composition of the present example is as follows (1 part = 1 kg):

[0037] Lobelia 20 parts, money grass 20 parts, chrysanthemum 15 parts, atractylodes 15 parts, atractylodes 10 parts, and cardamom kernel 10 parts.

[0038] Example 2 A traditional Chinese medicine composition for metastatic non-small cell lung cancer

[0039] The raw material composition of the traditional Chinese medicine composition of the present example is as follows (1 part = 1 kg):

[0040] Lobelia 20 parts, money grass 15 parts, chrysanthemum 15 parts, atractylodes 10 parts, atractylodes 10 parts, and cardamom kernel 6 parts.

[0041] Example 3 A traditional Chinese medicine composition for metastatic non-small cell lung cancer

[0042] The raw material composition of the traditional Chinese medicine composition of the present example is as follows (1 part = 1 kg):

[0043] Lobelia 15 parts, money grass 15 parts, chrysanthemum 10 parts, atractylodes 10 parts, atractylodes 9 parts, and cardamom kernel 6 parts.

[0044] Example 4 A capsule

[0045] Take the traditional Chinese medicine composition of Example 1, mix and grind into fine powder, pass through a 100 mesh sieve, and fill into capsules, 0.4 g of crude drug per particle, to obtain.

[0046] Dosage and method of use: 2-4 particles per time, three times a day, and take with warm water.

[0047] Example 5 A granule

[0048] Take the traditional Chinese medicine composition of Example 2, mix and grind into fine powder, pass through a 200 mesh sieve, add an appropriate amount of dextrin as a filler, granulate and press into granules, 6 g per bag, and pack, to obtain.

[0049] Dosage and method of use: 10 g per time, three times a day, and take with warm water.

[0050] Example 6 A tablet

[0051] Take the traditional Chinese medicine composition of Example 3, mix and grind into fine powder, pass through 80-100 mesh sieve, add dextrin and directly compress into tablets, 0.3g crude drug per tablet, and obtain.

[0052] Dosage and usage: 2-4 tablets each time, three times a day, and take with warm water.

[0053] Example 7 A pill

[0054] Take the traditional Chinese medicine composition of Example 1, mix and grind into fine powder, pass through 60-100 mesh sieve, add appropriate honey, and then prepare into small honey pills according to the conventional method, 3g per 10 pills, and obtain.

[0055] Dosage and usage: 3g each time, three times a day, and take with warm water.

[0056] Example 8 A decoction

[0057] The medicinal materials are weighed according to the following mass:

[0058] 15g of Lysimachia clethophylla, 15g of Lycopus lucidus, 10g of Chrysanthemum, 10g of Atractylodes, 9g of Atractylodes lancea, and 6g of Amomum kravanh.

[0059] Mix the medicinal materials, add 5-10 times the weight of water, heat to boiling, and then decoct for 25 minutes, filter while hot; add 5-10 times the weight of water to the residue, heat to boiling, decoct for 20-30 minutes, filter while hot, and combine the two times of medicinal liquid, and obtain. Take within 1 day in 2-3 times.

[0060] Experimental Example Pharmacodynamic experiment

[0061] 1. Materials

[0062] 1.1 Experimental drugs:

[0063] ①Traditional Chinese medicine composition: extract powder 1 (brown yellow, traditional Chinese medicine composition in the present application): content is 2.45g / g (crude drug amount / extract powder), extract powder 2 (brown yellow, control traditional Chinese medicine composition 1): content is 2.99g / g (crude drug amount / extract powder), extract powder 3 (brown yellow, control traditional Chinese medicine composition 2): content is 3.40g / g (crude drug amount / extract powder); concentration is shown in Table 1.

[0064] The preparation method of the above extract powder is:

[0065] Extract powder 1: take the traditional Chinese medicine composition described in Example 3, prepare the decoction according to the method described in Example 8, and then concentrate to 80℃ relative density 1.3-1.4, and obtain.

[0066] Infusion powder 2: Banlangen 10g, Guizhi 8g, Baoguo 8g, Chaihu 10g, Chuanxiong 8g, Mahuang 3g, Yujin 12g, Shanzhu 10g, Dangui 8g, Wuzhuyu 4g, Duhuo 6g; take each kind of medicinal material, prepare the decoction according to the method described in Example 8, and then concentrate to 80℃ relative density 1.3-1.4, to obtain.

[0067] Infusion powder 3: Banlangen 10g, Guizhi 8g, Baoguo 8g, Chaihu 10g, Chuanxiong 8g, Mahuang 3g, Yujin 12g, Shanzhu 10g, Dangui 8g, Wuzhuyu 4g, Duhuo 6g; take each kind of medicinal material, prepare the decoction according to the method described in Example 8, and then concentrate to 80℃ relative density 1.3-1.4, to obtain.

[0068] ②Paclitaxel, white powder, Shanghai Aladdin Bio-Chem Technology Co., Ltd., batch number J2126744, stored at 4℃. Prepare a 20.0mM stock solution with DMSO, dilute as needed, and store at -20℃.

[0069] 1.2 Experimental animals

[0070] ①Wild type AB strain zebrafish were bred by natural pair mating, a total of 300, 30 in each experimental group, 3dpf in age, used for maximum detection concentration (MTC) determination of the traditional Chinese medicine composition of the application.

[0071] ②Wild type AB strain zebrafish were bred by natural pair mating, a total of 1200, 30 in each experimental group, 2dpf in age, used for evaluation of the traditional Chinese medicine composition of the application on inhibiting non-small cell lung cancer metastasis.

[0072] The experimental animals were bred in fish water at 28℃ (water quality: 200mg of instant sea salt was added to each 1L of reverse osmosis water, conductivity was 480-510μS / cm; pH was 6.9-7.2; hardness was 53.7-71.6mg / L CaCO3), and the experimental animal license number was SYXK(Zhe)2022-0004. The breeding management met the requirements of international AAALAC certification.

[0073] 1.3 Experimental instruments:

[0074] ①Dissecting microscope (SZX7, OLYMPUS, Japan);

[0075] ②CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China);

[0076] ③Microinjector (IM300, NARISHIGE, Japan);

[0077] (4) Electrically focusing continuous zoom fluorescence microscope (AZ100, Nikon, Japan);

[0078] (5) Precision electronic balance (CP214, OHAUS, America);

[0079] (6) 6-well plate (Nest Biotech, Shanghai, China).

[0080] 2. Method

[0081] (1) Concentration group

[0082] Experiment 1 group model control group

[0083] Experiment 2 group positive control drug paclitaxel 1.00 pmol / tail

[0084] Experiment 3 group positive control drug paclitaxel 2.00 pmol / tail

[0085] Experiment 4 group positive control drug paclitaxel 4.00 pmol / tail

[0086] Experiment 5 group traditional Chinese medicine composition (extract powder 1) in the application 31.2 μg extract powder / mL

[0087] Experiment 6 group traditional Chinese medicine composition (extract powder 1) in the application 62.5 μg extract powder / mL

[0088] Experiment 7 group traditional Chinese medicine composition (extract powder 1) in the application 125 μg extract powder / mL

[0089] Experiment 8 group control traditional Chinese medicine composition 1 (extract powder 2) 31.2 μg extract powder / mL

[0090] Experiment 9 group control traditional Chinese medicine composition 1 (extract powder 2) 62.5 μg extract powder / mL

[0091] Experiment 10 group control traditional Chinese medicine composition 1 (extract powder 2) 125 μg extract powder / mL

[0092] Experiment 11 group control traditional Chinese medicine composition 2 (extract powder 3) 31.2 μg extract powder / mL

[0093] Experiment 12 group control traditional Chinese medicine composition 2 (extract powder 3) 62.5 μg extract powder / mL

[0094] Experiment 13 group control traditional Chinese medicine composition 2 (extract powder 3) 125 μg extract powder / mL

[0095] (2) Model making

[0096] The human non-small cell lung cancer (NCI-H460 and A549) cells were labeled with green fluorescent dye, and about 200 cells were transplanted into the yolk sac of zebrafish by microinjection to establish a zebrafish lung cancer tumor cell transplantation model.

[0097] (3) Experimental method

[0098] ①MTC determination

[0099] Randomly selected 330 wild type AB strain zebrafish of 3dpf in a six-well plate, 30 zebrafish were treated in each well (experimental group), and the drug concentration of the traditional Chinese medicine composition of the application was 125, 250, 500, 1000 and 2000 μg extract powder / mL, and a normal control group was set up, and the volume of each well (experimental group) was 3 mL. Incubate in a 35℃ incubator to 5dpf, count the death of zebrafish, and determine the MTC of the traditional Chinese medicine composition of the application on normal zebrafish according to the toxicity reaction and death of zebrafish.

[0100] ②Evaluation of the growth and metastasis inhibition effect of the traditional Chinese medicine composition of the application on non-small cell lung cancer (NCI-H460 and A549)

[0101] The human non-small cell lung cancer (NCI-H460 and A549) cells were labeled with green fluorescent dye, and about 200 cells were transplanted into the yolk sac of zebrafish by microinjection to establish a zebrafish lung cancer tumor cell transplantation model.

[0102]

[0103] Statistical analysis was performed using one-way ANOVA and Dunnett's T test, p<0.05 indicates significant difference, and representative experimental graphs are provided.

[0104] (4) Experimental results

[0105] ① Measurement of MTC

[0106] The normal zebrafish induced by the traditional Chinese medicine composition in the application at a concentration of 500 μg / mL had no more than 10% mortality and no obvious toxicity, and the tumor model zebrafish induced by the traditional Chinese medicine composition at a concentration of 250 μg / mL had no mortality and no toxicity phenotype, therefore, the MTC of the traditional Chinese medicine composition in the application and control traditional Chinese medicine composition 1 for inhibiting the growth of human metastatic non-small cell lung cancer in zebrafish was determined to be 250 μg / mL, and the MTC of control traditional Chinese medicine composition 2 for inhibiting the growth of human metastatic non-small cell lung cancer in zebrafish was determined to be 125 μg / mL. Details are shown in Table 1.

[0107] Table 1 Concentration-mortality results of normal zebrafish treated with the traditional Chinese medicine composition in the application (n=30)

[0108]

[0109] ② Growth inhibition of NCI-H460 transplanted tumor by the traditional Chinese medicine composition in the application

[0110] The fluorescence intensity of zebrafish NCI-H460 transplanted tumor at paclitaxel concentrations of 1.00 pmol / tail, 2.00 pmol / tail and 4.00 pmol / tail was 255176±15040, 181161±21683 and 144306±11680 pixels, respectively, and the high-dose group had a significant difference (p<0.01) compared with the model control group (27962.50±24644 pixels), and the tumor growth inhibition was 48.4%, indicating that the positive control drug can inhibit the growth of NCI-H460 transplanted tumor.

[0111] The fluorescence intensity of zebrafish NCI-H460 transplanted tumor at concentrations of 31.2, 62.5 and 125 μg / mL of the traditional Chinese medicine composition (extract powder 1) in the application was 275152±28973, 162435±19859 and 171974±18511 pixels, respectively, and the medium and high dose groups had extremely significant differences (p<0.01 and p<0.01, respectively) compared with the model control group (27962.50±24644 pixels), and the tumor growth inhibition was 41.91% and 38.50%, respectively. Details are shown in Table 2.

[0112] The fluorescence intensity of the zebrafish NCI-H460 transplanted tumor in the control traditional Chinese medicine composition 1 (infusion powder 2) 31.2, 62.5, 125 μg / mL concentration group was 234832 ± 30362.5, 202840 ± 24189, 152870 ± 9908 pixels, wherein the high-dose group had a significant difference (p < 0.01) compared with the model control group (27962.50 ± 24644 pixels), and the tumor growth inhibition was 45.34%.

[0113] The fluorescence intensity of the zebrafish NCI-H460 transplanted tumor in the control traditional Chinese medicine composition 2 (infusion powder 3) 31.2, 62.5, 125 μg / mL concentration group was 228483 ± 32757, 185791 ± 24387, 155179 ± 12587 pixels, wherein the high-dose group had a significant difference (p < 0.01) compared with the model control group (27962.50 ± 24644 pixels), and the tumor growth inhibition was 44.51%.

[0114] Therefore, the growth inhibition of the zebrafish NCI-H460 transplanted tumor in the high-dose group of the infusion powder 1, the high-dose group of the infusion powder 2, the high-dose group of the infusion powder 3 and the high-dose group of paclitaxel had a significant difference (p < 0.01) compared with the model group, indicating that the drugs all had the effect of inhibiting the growth of the zebrafish NCI-H460 transplanted tumor, and the traditional Chinese medicine composition in the application had a statistical difference compared with the model control group from the medium concentration. Details are shown in Table 2.

[0115] Table 2 Growth inhibition of the zebrafish NCI-H460 transplanted tumor by the traditional Chinese medicine composition of the application (n = 10)

[0116]

[0117] Note: Compared with the model control group, **p < 0.01, ***p ≤ 0.001.

[0118] ③ The metastasis inhibition effect of the traditional Chinese medicine composition of the application on NCI-H460 transplanted tumor

[0119] The NCI-H460 tumor cell metastasis distance of the zebrafish tail vein was 119950 ± 10289, 93602 ± 13052, 80031 ± 9542 at the concentration of 1.00 pmol / tail, 2.00 pmol / tail and 4.00 pmol / tail, respectively, and there was no statistical difference (p > 0.05) between each concentration group and the model control group (119209 ± 17587 pixels), indicating that the positive control drug at this concentration could not inhibit the metastasis of NCI-H460 transplanted tumor.

[0120] The NCI-H460 tumor cell metastasis distance of the traditional Chinese medicine composition (infusion powder 1) of the present application at 31.2, 62.5 and 125 μg / mL concentration groups was 71861±14093, 27380±3142 and 27755±2845 respectively, wherein the medium and high dose groups had extremely significant difference (p<0.001 and p<0.001 respectively) compared with the model control group (119209±17587 pixels), and the tumor metastasis inhibition effect was 77.03% and 76.72% respectively. See Table 3.

[0121] The NCI-H460 tumor cell metastasis distance of the control traditional Chinese medicine composition 1 (infusion powder 2) at 31.2, 62.5 and 125 μg / mL concentration groups was 112305±19751, 109459±21628 and 63297±7849 pixels respectively, and there was no statistical difference (p>0.05) compared with the model control group (119209±17587 pixels).

[0122] The NCI-H460 tumor cell metastasis distance of the control traditional Chinese medicine composition 2 (infusion powder 3) at 31.2, 62.5 and 125 μg / mL concentration groups was 76867±10817, 34809±3662 and 35008±5693 respectively, wherein the medium and high dose groups had extremely significant difference (p<0.001 and p<0.001 respectively) compared with the model control group (119209±17587 pixels), and the tumor metastasis inhibition effect was 70.80% and 70.63% respectively. Although the medium and high dose groups of the infusion powder 3 had significant difference (p<0.05) compared with the model group, the tumor inhibition rate of the medium and high dose groups was significantly lower than that of the traditional Chinese medicine composition (infusion powder 1) of the present application at the same concentration (p<0.05), indicating that the tumor growth inhibition effect of the traditional Chinese medicine composition (infusion powder 1) of the present application on the NCI-H460 transplanted tumor of zebrafish was obviously stronger than that of the control traditional Chinese medicine composition 2 (infusion powder 3).

[0123]

[0124] Therefore, the infusion powder 2 had the tendency to inhibit the metastasis of NCI-H460 transplanted tumor, but there was no statistical difference. Meanwhile, the tumor metastasis inhibition effect of the traditional Chinese medicine composition (infusion powder 1) of the present application on the NCI-H460 transplanted tumor of zebrafish was stronger than that of the control traditional Chinese medicine composition 1 (infusion powder 2) and the control traditional Chinese medicine composition 2 (infusion powder 3) and the positive control paclitaxel group. See Table 3.

[0125] Table 3 Tumor metastasis inhibition effect of the traditional Chinese medicine composition of the present application on the NCI-H460 transplanted tumor of zebrafish (n=10)

[0126]

[0127] ​

[0128] Note: compared with the model control group, **p<0.01, ***p≤0.001; compared with the same dose group of extract powder 1, # p<0.05.

[0129] 4. The growth inhibition effect of the traditional Chinese medicine composition of the present application on A549 transplanted tumors

[0130] The fluorescence intensity of zebrafish A549 transplanted tumors at paclitaxel concentrations of 1.00 pmol / tail, 2.00 pmol / tail and 4.00 pmol / tail was 326139±37347, 269894±24063 and 249623±25639 pixels, respectively, which was significantly different from that of the model control group (494191±44230 pixels) (p<0.01 and p<0.001, respectively, p<0.001), and the tumor growth inhibition was 34.01%, 45.39% and 49.49%, respectively, indicating that the positive control drug can significantly inhibit the growth of A549 transplanted tumors.

[0131] The fluorescence intensity of zebrafish A549 transplanted tumors at the concentrations of the traditional Chinese medicine composition (extract powder 1) of the present application (32.5, 62.5 and 125 μg / mL) was 408184±56037, 262577±31503 and 248242±20246 pixels, respectively, and the high-dose group was significantly different from the model control group (494191±44230 pixels) (p<0.01 and p<0.01, respectively), and the tumor growth inhibition was 46.87% and 49.77%, respectively, indicating that the traditional Chinese medicine composition of the present application can significantly inhibit the growth of zebrafish A549 transplanted tumors. See Table 4 for details.

[0132] The fluorescence intensity of zebrafish A549 transplanted tumors at the concentrations of the control traditional Chinese medicine composition 1 (extract powder 2) (32.5, 62.5 and 125 μg / mL) was 369323±39746, 338874±27826 and 331589±34753 pixels, respectively, and the high-dose group was significantly different from the model control group (494191±44230 pixels) (p<0.05 and p<0.05, respectively), and the tumor growth inhibition was 31.43% and 32.90%, respectively.

[0133] The fluorescence intensity of zebrafish A549 transplanted tumor in the control traditional Chinese medicine composition 2 (infusion powder 3) 32.5, 62.5 and 125 μg / mL concentration group was 405119 ± 39223, 299785 ± 31821, 196138 ± 29992 pixels, respectively, wherein the medium and high dose groups had significant difference with the model control group (494191 ± 44230 pixels) (p < 0.01 and p < 0.001, respectively), and the tumor growth inhibition was 39.34% and 60.31%, respectively.

[0134] Therefore, the traditional Chinese medicine composition (infusion powder 1) of the present application, the control traditional Chinese medicine composition 1 (infusion powder 2) and the control traditional Chinese medicine composition 2 (infusion powder 3) all have inhibitory effect on the growth of zebrafish A549 transplanted tumor. See Table 4 for details.

[0135] Table 4 Inhibitory effect of the traditional Chinese medicine composition of the present application on the growth of zebrafish A549 transplanted tumor (n = 10)

[0136]

[0137]

[0138] Compared with the model control group, **p < 0.01, ***p ≤ 0.001; compared with the same dose group of infusion powder 1, # p < 0.05.

[0139] 5. Inhibitory effect of the traditional Chinese medicine composition of the present application on A549 transplanted tumor

[0140] The tumor cell metastasis distance of zebrafish A549 tail vein in the paclitaxel 1.00 pmol / tail, 2.00 pmol / tail and 4.00 pmol / tail concentration group was 31673 ± 3467, 31137 ± 3553, 29941 ± 2953 pixels, which had extremely significant difference compared with the model control group (70006 ± 8870 pixels) (p < 0.001 and p < 0.001, respectively, p < 0.001), and the tumor metastasis inhibition was 54.76%, 55.52% and 57.23%, which indicated that the positive control drug could significantly inhibit the metastasis of A549 transplanted tumor.

[0141] The A549 tail vein tumor cell metastasis distances of the traditional Chinese medicine composition (infusion powder 1) of the present application at concentrations of 32.5, 62.5 and 125 μg / mL were 37236 ± 5113, 30906 ± 2519 and 23285 ± 2334 pixels, respectively, wherein the high and medium dose groups had extremely significant differences (p < 0.05 and p < 0.001, respectively) compared with the model control group (70006 ± 8870 pixels), and the tumor growth inhibition effects were 55.85% and 66.74%, respectively. This shows that the traditional Chinese medicine composition of the present application has a significant inhibitory effect on the metastasis of the A549 transplanted tumor of zebrafish. See Table 5.

[0142] The A549 tail vein tumor cell metastasis distances of the control traditional Chinese medicine composition 1 (infusion powder 2) at concentrations of 32.5, 62.5 and 125 μg / mL were 54377 ± 7528, 43264 ± 4009 and 31248 ± 2819 pixels, respectively, wherein the high dose group had a significant difference (p < 0.01) compared with the model control group (70006 ± 8870 pixels), and the tumor metastasis inhibition effect was 55.36%. However, the A549 transplanted tumor metastasis inhibition of each dose group of the infusion powder 2 was significantly lower than that of the traditional Chinese medicine composition of the present application at the same concentration (p < 0.05). This shows that the inhibitory effect of the traditional Chinese medicine composition (infusion powder 1) of the present application on the A549 transplanted tumor of zebrafish is significantly stronger than that of the control traditional Chinese medicine composition 1 (infusion powder 2). See Table 5.

[0143] The A549 tail vein tumor cell metastasis distances of the control traditional Chinese medicine composition 2 (infusion powder 3) at concentrations of 32.5, 62.5 and 125 μg / mL were 36060 ± 4114, 33075 ± 2991 and 26201 ± 3574 pixels, respectively, wherein the high dose group had significant differences (p < 0.05 and p < 0.001) compared with the model control group (70006 ± 8870 pixels), and the tumor metastasis inhibition effects were 52.75% and 62.57%. However, the A549 transplanted tumor metastasis inhibition of each dose group of the infusion powder 3 was significantly lower than that of the traditional Chinese medicine composition of the present application at the same concentration (p < 0.05), which shows that the inhibitory effect of the traditional Chinese medicine composition (infusion powder 1) of the present application on the A549 transplanted tumor of zebrafish is significantly stronger than that of the control traditional Chinese medicine composition 2 (infusion powder 3). See Table 5.

[0144] Table 5 Inhibitory effect of the traditional Chinese medicine composition of the present application on the metastasis of the A549 transplanted tumor of zebrafish (n = 10)

[0145]

[0146] Compared with the model control group, **p < 0.01, ***p ≤ 0.001; compared with the same dose group of the infusion powder 1, # p < 0.05.

[0147] In summary, the present application provides a traditional Chinese medicine composition which can be used for treating metastatic non-small cell lung cancer. The composition comprises Lobelia chinensis, Atractylodes macrocephala, Herba Lysimachiae, Chrysanthemum, Atractylodes lancea, and Amomum kravanh, which can clear heat and resolve toxins, and invigorate the spleen and eliminate dampness. Pharmacological experiments prove that the traditional Chinese medicine composition of the present application can significantly inhibit the growth and metastasis of human non-small cell lung cancer (NCI-H460 and A549) transplanted tumors. The present application provides a new clinical treatment option for patients with metastatic non-small cell lung cancer and doctors.

Claims

1. A traditional Chinese medicine composition for metastatic non-small cell lung cancer, comprising Lobelia chinensis, Atractylodes lancea, Amomum villosum seeds, Atractylodes macrocephala, Lysimachia chinensis, and Chrysanthemum, wherein the weight proportions of each medicinal material are as follows: 10-20 parts of Lobelia chinensis, 10-20 parts of Lysimachia chinensis, 5-15 parts of chrysanthemum, 5-15 parts of Atractylodes macrocephala, 5-10 parts of Atractylodes lancea, and 5-10 parts of Amomum villosum seeds.

2. The Chinese medicine composition according to claim 1, characterized in that The weight parts of each medicinal material are: 15 parts of Herba Lobeliae, 15 parts of Herba Lysimachiae, 10 parts of Flos Chrysanthemi, 10 parts of Rhizoma Atractylodis Macrocephalae, 9 parts of Rhizoma Atractylodis, 6 parts of Semen Amomi.

3. A medicine, the raw materials of which include the Chinese medicine composition according to claim 1 or 2.

4. The drug according to claim 3, characterized in that Pharmaceutically acceptable excipients are also included.

5. The medicine according to claim 3 or 4, characterized in that The drug is any clinically acceptable preparation.

6. The drug according to claim 5, characterized in that The drug is an oral preparation.

7. The drug according to claim 6, characterized in that The oral preparation is selected from decoction, powder, capsule, tablet, honey pill, water-honey pill, water pill, concentrated pill, paste pill, wax pill, granule, oral liquid or dripping pill.

8. The drug according to claim 7, characterized in that The oral preparation is a decoction or a granule.

9. A method for preparing the drug according to any one of claims 3 to 8, comprising preparing the medicinal materials in the proportions by weight as recited in claim 1 or 2, and preparing a clinically acceptable preparation according to conventional methods in the art, with or without the addition of pharmaceutically acceptable excipients.

10. Use of the traditional Chinese medicine composition according to claim 1 or 2, the medicine according to any one of claims 3 to 8, or the medicine prepared according to the preparation method according to claim 9 in the preparation of a medicine for treating non-small cell lung cancer.

11. Use of the traditional Chinese medicine composition according to claim 1 or 2, the medicine according to any one of claims 3 to 8, or the medicine prepared according to the preparation method according to claim 9 in preparing a medicine for treating metastatic non-small cell lung cancer.

12. Use of the traditional Chinese medicine composition according to claim 1 or 2, the medicine according to any one of claims 3 to 8, or the medicine prepared according to the preparation method according to claim 9 in the preparation of a medicine for treating unresectable metastatic non-small cell lung cancer, metastatic non-small cell lung cancer after surgery, or metastatic non-small cell lung cancer that has recurred after surgery.

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