Traditional Chinese medicine composition for inhibiting lung cancer metastasis and application thereof

Abstract

The invention relates to the technical field of traditional Chinese medicines, in particular to a traditional Chinese medicine composition for inhibiting lung cancer metastasis and application thereof. The invention provides a traditional Chinese medicine composition and a medicine for inhibiting lung cancer metastasis. The traditional Chinese medicine composition comprises folium isatidis, selfheal, radix bupleuri, purslane, dandelion, cassia twig, semen raphani and liquorice. The traditional Chinese medicine composition and the medicine have the effect of preventing postoperative recurrence and metastasis of the lung cancer, and the life quality of a patient can be improved; the technical defect of many adverse reactions of western medicines can be overcome, and meanwhile, the pharmaceutical composition has the advantages of small side effects, long drug effect persistence and multiple drug targets. The invention also provides a preparation method of the traditional Chinese medicine composition for inhibiting lung cancer metastasis, and according to the method, the raw materials are subjected to water extraction and then treated by an enzyme preparation and a fermentation inoculant, so that the dissolution rate of effective components can be improved.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to a traditional Chinese medicine composition for inhibiting lung cancer metastasis and application thereof. Background Art

[0002] Primary bronchogenic carcinoma, or lung cancer for short, refers to a malignant tumor in which the epithelial cells of the alveolar wall, bronchial or bronchiolar mucosa proliferate without restriction. In 2020, the overall 5-year relative survival rate of cancer was 67%, while the survival rate of lung cancer was as low as 19%. The 5-year relative survival rate of patients with early lung cancer was 57%, while that of patients with advanced disease was only 5%. Lung cancer is currently the most common malignant tumor with the highest mortality rate. Current medical measures still make it difficult to diagnose lung cancer early, and most patients are already in the middle and late stages when they seek medical treatment or are diagnosed, which makes the treatment of lung cancer very difficult. Surgical treatment is suitable for non-small cell lung cancer (NSCLC) below grade IIIA that occurs on one side. At present, the failure rate of lung cancer surgery can reach 4.4%, and most patients can only receive radiotherapy or chemotherapy, but due to the high toxicity of radiotherapy and chemotherapy, bone marrow suppression and low immunity can be easily caused.

[0003] Although treatment methods have been continuously updated and treatment plans have been continuously optimized in the past decade, the 5-year overall survival rate of lung cancer patients is still only 14%. At present, clinical studies have revealed that postoperative recurrence and metastasis are one of the main reasons why the overall 5-year survival rate of lung cancer cannot be improved. Therefore, the development of a Chinese medicine composition that can effectively inhibit lung cancer metastasis is of great significance to improving the survival rate of lung cancer patients. Summary of the invention

[0004] The purpose of the present invention is to provide a Chinese medicine composition for inhibiting lung cancer metastasis and its application, so as to solve the problems existing in the above-mentioned prior art. The Chinese medicine composition provided by the present invention can effectively inhibit lung cancer metastasis and improve the survival rate of lung cancer patients.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The present invention provides a traditional Chinese medicine composition for inhibiting lung cancer metastasis, comprising the following components in parts by weight:

[0007] 10-13 parts of Folium Isatidis, 15-20 parts of Herba Prunellae, 12-15 parts of Radix Bupleuri, 5-7 parts of Herba Portulacae, 10-15 parts of Herba Taraxaci, 5-8 parts of Herba Cinnamomi, 5-10 parts of Herba Radix Raphani and 1-5 parts of Radish Root.

[0008] Preferably, the following components are included by weight:

[0009] 12 parts of Folium Isatidis, 18 parts of Herba Prunellae, 13 parts of Radix Bupleuri, 6 parts of Portulaca Oleracea, 14 parts of Herba Taraxaci, 7 parts of Cinnamomum cassia, 8 parts of Semen Raphani, and 3 parts of Radish Root;

[0010] Or, 10 parts of Folium Isatidis, 20 parts of Herba Prunellae, 12 parts of Radix Bupleuri, 7 parts of Herba Portulacae, 10 parts of Herba Taraxaci, 8 parts of Herba Cinnamomi, 10 parts of Semen Radix Raphani and 5 parts of Radix Glycyrrhizae.

[0011] The present invention provides application of the above-mentioned traditional Chinese medicine composition in preparing medicine for inhibiting lung cancer metastasis.

[0012] The present invention provides a medicine for inhibiting lung cancer metastasis, and the medicine comprises the above-mentioned traditional Chinese medicine composition.

[0013] Preferably, the drug further comprises a pharmaceutically acceptable excipient.

[0014] Preferably, the dosage form of the drug is decoction, pill, capsule, tablet, powder or granule.

[0015] The present invention provides a method for preparing the above-mentioned medicine, comprising the following steps:

[0016] After mixing the isatis indigo plant, the selfheal, the bupleurum, the purslane, the dandelion, the cassia twig, the radish seed and the liquorice, performing a first water extraction to obtain a first filter residue and a first filtrate; performing a second water extraction on the first filter residue to obtain a second filter residue and a second filtrate; combining the first filtrate and the second filtrate to obtain a water extract;

[0017] Performing enzymatic hydrolysis on the second filter residue to obtain an enzymatic hydrolysis solution and an enzymatic hydrolysis residue;

[0018] fermenting the enzymatic hydrolysis residue to obtain a fermentation liquid;

[0019] The water extract, the enzymatic hydrolyzate and the fermentation liquid are mixed evenly to obtain the medicine.

[0020] Preferably, during the first water extraction, the mass ratio of the total mass of the isatis indigo plant, the selfheal, the bupleurum, the purslane, the dandelion, the cassia twig, the radish seed and the liquorice to water is 1:5, and the time is 1.5 hours;

[0021] And / or, during the second water extraction, the mass ratio of the first filter residue to water is 1:2, and the time is 2 hours.

[0022] Preferably, during the enzymatic hydrolysis, the enzyme preparation used is a composite enzyme preparation containing trypsin and neutral protease; the amount of the enzyme preparation used is 0.5% of the mass of the second filter residue;

[0023] And / or, the enzymatic hydrolysis time is 45 min and the temperature is 45°C.

[0024] Preferably, during the fermentation, the fermentation bacteria used include Bacillus megaterium, Bacillus licheniformis and Lactococcus lactis; the amount of the fermentation bacteria used is 3% of the mass of the enzymatic residue;

[0025] And / or, the fermentation time is 14 hours and the temperature is 38°C.

[0026] The present invention discloses the following technical effects:

[0027] The present invention provides a Chinese medicine composition and medicine for inhibiting lung cancer metastasis. In the composition, Prunella vulgaris dissipates stagnation and reduces swelling, Isatis indigotica and Portulaca oleracea clear away heat and detoxify, and are monarch drugs. The three together play the effects of clearing heat, dissipating stagnation and reducing swelling; Bupleurum soothes the liver and relieves depression, Radish seeds descend qi and resolve phlegm, and are minister drugs; Cinnamon twig assists yang and transforms qi, and calms and descends qi, Taraxacum officinale reduces swelling and dissipates stagnation, and assists the monarch drug, and is a guiding drug; Licorice is sweet and flat in nature and flavor, relieves bitter and cold nature and can reconcile various drugs, and is a guiding drug. The compatibility of various drugs together plays the effects of dissipating stagnation, reducing swelling, clearing heat and removing turbidity. The results of the embodiments of the present invention show that the Chinese medicine composition and medicine have the effect of preventing recurrence and metastasis of lung cancer after surgery, and can improve the quality of life of patients; they can also overcome the technical shortcomings of many adverse reactions of Western medicines, and have the advantages of small side effects, long duration of drug effect and multiple drug targets.

[0028] The present invention also provides a method for preparing a drug for inhibiting lung cancer metastasis, wherein the method extracts the raw material with water and then processes it with enzyme preparations and fermentation bacteria, thereby increasing the dissolution rate of the active ingredient. Therefore, the drug prepared by the method provided by the present invention has better efficacy. DETAILED DESCRIPTION

[0029] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0030] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0031] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0032] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0033] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0034] Isatis indigofera is a biennial herbaceous plant of the Cruciferae family, mainly produced in Hebei, Shaanxi, and Jiangsu. It is harvested 2-3 times in summer and autumn and used fresh or dried. Isatis indigofera is bitter and cold in nature; it enters the heart and stomach meridians. Isatis indigofera has the effects of clearing away heat and detoxifying, cooling blood and removing spots. It is mainly used to treat heat entering the blood, warm toxin spots, throat paralysis, mouth sores, and mumps erysipelas.

[0035] Prunella vulgaris is also known as wheat ear prunella, iron wire prunella (Yunnan series), wheat prunella, iron wire prunella (Yunnan Materia Medica), evening sentence, Naidong (benjing), etc. Prunella vulgaris is a perennial herb with creeping rhizomes and fibrous roots on the nodes. The stem is up to 30cm tall, with many branches at the base and light purple. The calyx is bell-shaped, the filaments are slightly flat, the style is slender, the tip lobes are awl-shaped, and they are bent outward. The flower disk is nearly flat-topped. The small nuts are yellow-brown, and the flowering period is from April to June, and the fruiting period is from July to October. Prunella vulgaris grows in wetlands in mountain valleys or wet grass on both sides of the river bank, wasteland, and roadsides. It is widely distributed throughout China, with Henan, Anhui, Jiangsu, and Hunan as the main production areas. Prunella vulgaris has strong adaptability and rarely suffers from diseases and pests throughout the growth process. Prunella vulgaris tastes spicy and bitter, and is cold in nature; it enters the liver and gallbladder meridians. It has the effects of clearing heat and improving eyesight, dispersing knots and reducing swelling, and is mainly used to treat red, swollen and painful eyes, headaches, etc.

[0036] Bupleurum was first recorded in Shennong's Herbal Classic and listed as a top-grade herb. The herbal classics of various dynasties have many descriptions of the plant morphology of Bupleurum. For example, the Illustrated Classic of Materia Medica records: "(Bupleurum) is now found in Guan, Shaanxi, Jianghu, and nearby roads, and the ones from Yinzhou are the best. Seedlings grow in February, very fragrant, with purple stems and slightly purple leaves like bamboo leaves... Yellow flowers bloom in July... The roots are red, similar to Peucedanum but stronger. The reed head has red hair like a mouse's tail, and the ones with a single nest and a long one are the best. The roots are harvested in February and August." Bupleurum tastes bitter and pungent, and is slightly cold in nature; it enters the liver and gallbladder meridians. It has the effects of relieving fever, soothing the liver and relieving depression, and raising yang qi. It is mainly used to treat fever caused by superficial symptoms, Shaoyang syndrome, liver depression and qi stagnation, qi deficiency and sinking, organ prolapse, and relieving fever and stopping malaria.

[0037] Purslane is an annual herb with no hair on the whole plant. The stem is prostrate and spreads on the ground, with branches light green or dark red. The leaves are alternate, flat, thick, and horse-tooth-shaped, dark green on the upper side and light green or dark red on the lower side; the petioles are short and thick. The flowers are sessile and bloom at noon; the bracts are leaf-shaped; the sepals are green and helmet-shaped; the petals are yellow and obovate; the stamens and anthers are yellow; the ovary is glabrous. The capsule is ovoid; the seeds are small, obliquely spherical, dark brown, and shiny. The flowering period is from May to August, and the fruiting period is from June to September. It is produced in all parts of China, north and south, and likes fertile soil. It is drought-resistant and waterlogged, and has strong vitality. It grows in vegetable gardens, farmlands, and roadsides. It is a common weed in the fields and is widely distributed in temperate and tropical regions around the world. Its seeds have the effect of improving eyesight; it can also be used as veterinary medicine and pesticides; the tender stems and leaves can be used as vegetables, and are also very good feed. Purslane tastes sour and is cold in nature; it enters the liver and large intestine meridians. It has the effects of clearing away heat and detoxifying, cooling blood and stopping bleeding, and stopping dysentery. It is mainly used to treat heat-toxic bloody dysentery, heat-toxic sores, metrorrhagia and bloody stools.

[0038] Dandelion is also known as Huanghuadiding, Dandelion, Huahualang, etc. Dandelion is a perennial herbaceous plant of the Asteraceae family and the genus Taraxacum. The root is conical, the surface is brown and wrinkled, the leaf margins are sometimes wavy or pinnate, the base gradually narrows into a petiole, the petiole and main veins are often reddish purple, the upper part of the scape is purple-red, densely covered with spider-like white long hairs; the inflorescence is head-shaped, the involucre is bell-shaped, the achenes are dark brown, the long pappus is white, and the flowering and fruiting period is from April to October. There are pompoms formed by white pappus on the seeds. Dandelion tastes bitter and sweet, and is cold in nature; it enters the liver and stomach meridians. It has the effects of clearing away heat and detoxification, reducing swelling and dispersing knots, and promoting dampness and relieving stranguria. It is mainly used to treat carbuncle, carbuncle, breast carbuncle, internal carbuncle, hot stranguria, damp-heat jaundice, and clearing the liver and improving eyesight.

[0039] Cinnamon twig, also known as Liugui, is the dried young branches of cinnamon bark of the Lauraceae family. It is harvested in spring and summer, leaves are removed, and it is dried in the sun or sliced ​​and dried in the sun. It is long cylindrical, branched, 30-75cm long, and 0.3-1cm in diameter at the thick end. The surface is reddish brown to brown, with longitudinal ridges, fine wrinkles, and small pimple-like leaf marks, branch marks, and bud marks, and the lenticels are dotted. It is hard and brittle and easy to break. The slices are 2-4mm thick, the cross-section of the skin is reddish brown, and the pith is slightly square. It has a unique aroma, sweet and slightly spicy, and the skin tastes stronger. It is mainly produced in Guangxi, Guangdong, Yunnan and other places. Cinnamon twig tastes spicy and sweet, and is warm in nature. It enters the heart, lung, and bladder meridians. It has the effects of sweating and relieving muscles, warming and unblocking meridians, helping yang and transforming qi, and calming and descending qi. It is often used for colds, cold pain in the abdomen, amenorrhea due to blood cold, joint pain, phlegm and fluid retention, edema, and palpitations.

[0040] Radish seeds are also known as radish seeds, white radish seeds, and cabbage seeds. In summer, when the fruits are ripe, the plants are harvested, dried, the seeds are rubbed out, impurities are removed, and then dried again to finally obtain radish seeds. Radish seeds are spicy and sweet, and are neutral in nature; they enter the lung, spleen, and stomach meridians. They have the effects of digesting food, eliminating bloating, and lowering qi and resolving phlegm. They are mainly used to treat food stagnation, cough, asthma, excessive phlegm, and chest tightness and loss of appetite.

[0041] Licorice, also known as Guolao, sweet grass, Ural licorice, and sweet root, is a perennial herb of the Leguminosae family and the genus Glycyrrhiza. It has thick roots and rhizomes and is a tonic Chinese herbal medicine. The medicinal parts are the roots and rhizomes. The medicinal material characteristics are that the roots are cylindrical, 25-100cm long, and 0.6-3.5cm in diameter. The outer skin is loose and tight, and the surface is reddish brown or grayish brown. The rhizome is cylindrical, with bud marks on the surface and pith in the middle of the cross section. The smell is slight, and the taste is sweet and special. Functions and indications include clearing away heat and detoxification, expectoration and cough relief, and abdominal pain. It likes a dark and humid, dry climate with long sunshine and low temperature. Licorice grows mostly in arid and semi-arid desert grasslands, desert edges, and loess hilly areas. The roots and rhizomes are used for medicinal purposes. Licorice is flat in nature and sweet in taste; it enters the heart, lung, spleen, and stomach meridians. It has the functions of tonifying the spleen and replenishing qi, anti-inflammatory and sterilization, enhancing immunity, relieving pain, and harmonizing various medicines. Licorice can enhance immunity, fight inflammation, resist gastric ulcers, resist arrhythmias and regulate blood sugar, etc.

[0042] Example 1

[0043] A drug for inhibiting lung cancer metastasis, the raw materials of which include the following components in parts by weight:

[0044] 12 parts of Folium Isatidis, 18 parts of Herba Prunellae, 13 parts of Radix Bupleuri, 6 parts of Herba Portulacae, 14 parts of Herba Taraxaci, 7 parts of Herba Cinnamomi, 8 parts of Semen Radix Raphani and 3 parts of Radish.

[0045] The preparation method of the drug is as follows:

[0046] (1) After weighing the raw materials according to the raw material formula, rinse them with clean water once, filter out dust and impurities, and then extract them with water at a mass ratio of 1:5 and 100° C. for 2 hours to obtain a first filtrate and a first filter residue; extract the first filter residue with water at a mass ratio of the first filter residue to water of 1:2, at a temperature of 100° C., for 2 hours to obtain a second filtrate and a second filter residue; combine the first filtrate and the second filtrate to obtain a water extract;

[0047] (2) mixing the second filter residue with water in a mass ratio of 1:1, then adding an enzyme preparation, and performing enzymolysis at 45° C. for 45 min to obtain an enzymatic solution and an enzymatic residue; the amount of the enzyme preparation is 0.5% of the mass of the second filter residue, the enzyme preparation comprises trypsin and neutral protease, the enzyme activity of the trypsin in the enzyme preparation is 8000 U / mL, and the enzyme activity of the neutral protease is 5000 U / mL;

[0048] (3) The enzymatic residue and water were mixed in a mass ratio of 1:1, and then a fermentation agent was added, and fermented at 40°C for 18 hours to obtain a fermentation liquid, wherein the amount of the fermentation agent was 3% of the mass of the enzymatic residue, and the fermentation agent consisted of Bacillus megaterium, Bacillus licheniformis and Lactococcus lactis, and the number of viable bacteria of Bacillus megaterium in the fermentation agent was 1×10 6 CFU / mL, the number of viable bacteria of Bacillus licheniformis is 1×10 6 CFU / mL, the number of viable Lactococcus lactis is 1×10 5 CFU / mL;

[0049] (3) The water extract, the enzymatic hydrolyzate and the fermentation liquid are mixed evenly to obtain a drug for inhibiting lung cancer metastasis.

[0050] Example 2

[0051] A drug for inhibiting lung cancer metastasis, the raw materials of which include the following components in parts by weight:

[0052] 10 parts of Folium Isatidis, 20 parts of Herba Prunellae, 12 parts of Radix Bupleuri, 7 parts of Herba Portulacae, 10 parts of Herba Taraxaci, 8 parts of Herba Cinnamomi, 10 parts of Herba Radix Radishes and 5 parts of Radix Glycyrrhizae.

[0053] The preparation method of the drug is the same as that of Example 1.

[0054] Comparative Example 1

[0055] 12 parts of Folium Isatidis, 18 parts of Herba Prunellae, 13 parts of Radix Bupleuri, 14 parts of Herba Taraxaci, 7 parts of Rhizoma Cinnamomi, 8 parts of Semen Radix Raphani and 3 parts of Radix Glycyrrhizae.

[0056] The preparation method of the drug is the same as that of Example 1.

[0057] Comparative Example 2

[0058] 12 parts of Folium Isatidis, 18 parts of Herba Prunellae, 13 parts of Radix Bupleuri, 6 parts of Puffball, 14 parts of Taraxaci, 7 parts of Cinnamomum cassia, 8 parts of Semen Raphani and 3 parts of Radish.

[0059] The preparation method of the drug is the same as that of Example 1.

[0060] Comparative Example 3

[0061] The components are the same as those in Example 1.

[0062] The preparation method of the drug is as follows:

[0063] (1) After weighing the raw materials according to the raw material formula, rinse them with clean water once, filter out dust and impurities, and then extract them with water at a mass ratio of 1:5 and 100° C. for 2 hours to obtain a first filtrate and a first filter residue; extract the first filter residue with water at a mass ratio of the first filter residue to water of 1:2, at a temperature of 100° C., for 2 hours to obtain a second filtrate and a second filter residue; combine the first filtrate and the second filtrate to obtain a water extract;

[0064] (2) The second filter residue and water were mixed in a mass ratio of 1:1, and then a fermentation agent was added, and fermented at 40° C. for 18 h to obtain a fermentation liquid, wherein the amount of the fermentation agent was 3% of the mass of the second filter residue, and the fermentation agent consisted of Bacillus megaterium, Bacillus licheniformis and Lactococcus lactis, and the number of viable bacteria of Bacillus megaterium in the fermentation agent was 1×10 6 CFU / mL, the number of viable bacteria of Bacillus licheniformis is 1×10 6 CFU / mL, the number of viable Lactococcus lactis is 1×10 5 CFU / mL;

[0065] (3) The water extract and the fermentation liquid are mixed evenly to obtain a drug for inhibiting lung cancer metastasis.

[0066] Comparative Example 4

[0067] The components are the same as those in Example 1.

[0068] The preparation method of the drug is as follows:

[0069] (1) After weighing the raw materials according to the raw material formula, rinse them with clean water once, filter out dust and impurities, and then extract them with water at a mass ratio of 1:5 and 100° C. for 2 hours to obtain a first filtrate and a first filter residue; extract the first filter residue with water at a mass ratio of the first filter residue to water of 1:2, at a temperature of 100° C., for 2 hours to obtain a second filtrate and a second filter residue; combine the first filtrate and the second filtrate to obtain a water extract;

[0070] (2) mixing the second filter residue with water in a mass ratio of 1:1, then adding an enzyme preparation, and performing enzymolysis at 45° C. for 45 min to obtain an enzymatic solution and an enzymatic residue; the amount of the enzyme preparation is 0.5% of the mass of the second filter residue, the enzyme preparation comprises trypsin and neutral protease, the enzyme activity of the trypsin in the enzyme preparation is 8000 U / mL, and the enzyme activity of the neutral protease is 5000 U / mL;

[0071] (3) The water extract and the enzymatic hydrolyzate are mixed evenly to obtain a drug for inhibiting lung cancer metastasis.

[0072] Experimental Example 1 In vitro experiment

[0073] 1. Cell lines

[0074] Human non-small cell lung cancer H460 was purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences.

[0075] 2. Medication

[0076] The drugs for inhibiting lung cancer metastasis prepared in Examples 1-2 and Comparative Examples 1-4.

[0077] 3. Cell proliferation assay

[0078] H460 cells were subcultured and used for experiments when the cells grew to the logarithmic phase. The drugs for inhibiting lung cancer metastasis prepared in Examples 1-2 and Comparative Examples 1-4 were prepared into DMSO solutions with a concentration of 30 mL / mL. Lung cancer cells in the logarithmic growth phase were collected and prepared into a cell concentration of 3×10 4 cells / mL, 100 μL per well (3000 cells / well) was inoculated into a 96-well plate, and three 96-well plates were repeated. The zero well was set and the cells were cultured in a cell culture incubator at 37°C, 5% CO2, and saturated humidity.

[0079] After 24 hours, the cultured cells were taken out, the culture medium was discarded, and culture medium containing different concentrations of drugs (0, 12.5, 50, 100 and 150 μL / mL) was added respectively, and the cells were placed in a humidity incubator at 37°C, 5% CO2, and saturated humidity for 24 hours, 48 ​​hours, and 72 hours respectively. After reaching the end of the culture time point, the culture was terminated, 10 μL of CCK-8 solution was added to each well, and the incubation was continued until the OD value of the control group was equal to 1 at a detection wavelength of 450 nm; 450 nm was set as the detection wavelength, and the optical density value (OD value) in each well was detected, and the cell inhibition rate was calculated. The results are shown in Table 1. As shown in Table 1, the inhibitory effect of the drugs (Examples 1 and 2) provided by the present invention on H460 cells is significantly better than that of the drugs prepared by Comparative Examples 1-4.

[0080] Table 1 Inhibition rate of H460 cells

[0081]

[0082] Experimental Example 2 In vivo experiment

[0083] 1. Animals

[0084] 40 SPF BALB / c-nu / nu nude mice, 6-7 weeks old, weighing 20-22 g, male. The bedding, drinking water, standard feed and other items in contact with animals were sterilized by high pressure. They were purchased from Shanghai Xipu-Bikai Laboratory Animal Co., Ltd., license number: SCXK (Shanghai) 2018-0006.

[0085] 2. Medication

[0086] The drug for inhibiting lung cancer metastasis prepared in Example 1.

[0087] 3. Animal modeling

[0088] Drug intervention was started after one week of adaptive feeding. 40 mice were randomly divided into a drug group for inhibiting lung cancer metastasis and a normal saline control group, with 20 mice in each group.

[0089] 3.1 Modeling

[0090] After one week of feeding, each mouse was injected with 1×10 5 cells / mLH460 cells, when all H460 lung cancer subcutaneous transplanted tumors were formed and grew to a diameter of about 1 cm, which took an average of 15 days, surgical resection was performed to obtain a lung cancer metastasis mouse model (NCI-H460 lung cancer subcutaneous transplanted tumor nude mouse). On the second day after surgery, after in vivo imaging was observed to have no surgical residue, the lung cancer metastasis mouse model was randomly divided into 20 treatment groups and 20 control groups, and physiological saline and drug intervention were given respectively from the second day after surgery.

[0091] 3.2 Grouping and Dosing

[0092] 1) Treatment group: the drug for inhibiting lung cancer metastasis prepared in Example 1 (concentration of 50 μL / mL); intervention time and period: starting from the second day after surgery, the drug was gavaged, with a gavage volume of 0.1 mL / mouse per time, once every 7 days until 7 weeks after surgery;

[0093] 2) Control group: Normal saline group (NS): 0.9% normal saline, Shanghai Rongbai Biotechnology Co., Ltd., specification 100mL / bottle; Intervention time and cycle: Starting from the second day after surgery, gavage was performed, with each gavage volume of 0.1mL / mouse, once every 7 days until 7 weeks after surgery.

[0094] 3.3 Small Animal In vivo Imaging Observation

[0095] After modeling, nude mice bearing NCI-H460 lung cancer subcutaneous transplanted tumors were intraperitoneally injected with 150 mg / kg D-luciferin potassium (10 mg / mL dissolved in sterile water for injection) 1 day before surgery and 2 days after surgery, and 1, 2, 3, 4, 5, 6, and 7 weeks after surgery. After the injection, wait for 10 minutes for its metabolism in the body. After isoflurane gas anesthesia, the mice were placed in an imaging chamber to dynamically observe the tumor growth and metastasis.

[0096] At the same time, the lung metastasis inhibition rate was calculated. The calculation formula of the lung metastasis inhibition rate was: lung metastasis inhibition rate = (1-the number of tumors in the treatment group / the number of tumors in the control group) × 100%.

[0097] 4. Experimental results

[0098] The in vivo imaging system was used to detect that tumor cells with growth activity showed luminescence imaging when they absorbed D-luciferin potassium substrate. On the third day after surgery, no tumor residue was found in the surgical site of nude mice with NCI-H460 lung cancer subcutaneously transplanted tumors. The effect of in vivo imaging on disease-free survival time after surgery was not observed.

[0099] The in vivo imaging system was used to detect the rate of tumor recurrence and metastasis in nude mice in each group after surgery. The results are shown in Table 2.

[0100] Table 2 Postoperative tumor recurrence and metastasis rate in nude mice (%)

[0101]

[0102] As shown in Table 2, the recurrence-free metastasis rates of the treatment group at 7-49 days were 100%, 95%, 95%, 80%, 70%, 65% and 40% respectively; the recurrence-free metastasis rates of the control group at 7-49 days were 75%, 55%, 50%, 50%, 25%, 5% and 0% respectively. It can be seen that the drug provided by the present invention can effectively prevent the recurrence and metastasis of tumors.

[0103] Mice with recurrence and metastasis observed by in vivo imaging were sacrificed at week 7, and 8 mice were selected from each group to observe the number of lung metastases under a microscope. The results are shown in Table 3.

[0104] Table 3 Occurrence of lung metastasis

[0105] deal with N Lung metastasis Inhibition rate (%) Control group 8 9 - Treatment Group 8 2 78％

[0106] Note: The data in the table is the average of 8 numbers. If there are decimals, they will be rounded off.

[0107] As shown in Table 3, the control group had more lung metastases. The number of lung metastases in both lungs was significantly higher in the control group (9) than in the treatment group (2). The lung metastasis inhibition rate in the treatment group was 78%.

[0108] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A Chinese medicine composition for inhibiting lung cancer metastasis, characterized in that: The composition comprises the following components in parts by weight: 10-13 parts of Folium Isatidis, 15-20 parts of Herba Prunellae, 12-15 parts of Radix Bupleuri, 5-7 parts of Herba Portulacae, 10-15 parts of Herba Taraxaci, 5-8 parts of Herba Cinnamomi, 5-10 parts of Herba Radix Raphani and 1-5 parts of Radish Root.

2. The Chinese medicine composition according to claim 1, characterized in that: The composition comprises the following components in parts by weight: 12 parts of Folium Isatidis, 18 parts of Herba Prunellae, 13 parts of Radix Bupleuri, 6 parts of Portulaca Oleracea, 14 parts of Herba Taraxaci, 7 parts of Cinnamomum cassia, 8 parts of Semen Raphani, and 3 parts of Radish Root; Or, 10 parts of Folium Isatidis, 20 parts of Herba Prunellae, 12 parts of Radix Bupleuri, 7 parts of Herba Portulacae, 10 parts of Herba Taraxaci, 8 parts of Herba Cinnamomi, 10 parts of Semen Radix Raphani and 5 parts of Radix Glycyrrhizae.

3. Use of the Chinese medicine composition according to claim 1 or 2 in the preparation of a medicament for inhibiting lung cancer metastasis.

4. A drug for inhibiting lung cancer metastasis, characterized in that: The medicine comprises the Chinese medicine composition according to claim 1 or 2.

5. The drug according to claim 4, characterized in that The drug also includes pharmaceutically acceptable excipients.

6. The drug according to claim 5, characterized in that The dosage form of the medicine is decoction, pill, capsule, tablet, powder or granule.

7. The method for preparing the drug according to claim 4, characterized in that: The following steps are involved: The isatis indigo plant, the selfheal, the bupleurum, the purslane, the dandelion, the cassia twig, the radish seed and the liquorice are mixed, and then subjected to a first water extraction to obtain a first filter residue and a first filtrate; Performing a second water extraction on the first filter residue to obtain a second filter residue and a second filtrate; combining the first filtrate and the second filtrate to obtain a water extract; Performing enzymatic hydrolysis on the second filter residue to obtain an enzymatic hydrolysis solution and an enzymatic hydrolysis residue; fermenting the enzymatic hydrolysis residue to obtain a fermentation liquid; The water extract, the enzymatic hydrolyzate and the fermentation liquid are mixed evenly to obtain the medicine.

8. The preparation method according to claim 7, characterized in that: During the first water extraction, the mass ratio of the total mass of the isatis indigo plant, the selfheal, the bupleurum, the purslane, the dandelion, the cassia twig, the radish seed and the liquorice to water is 1:5, and the extraction time is 1.5 hours; And / or, during the second water extraction, the mass ratio of the first filter residue to water is 1:2, and the time is 2 hours.

9. The preparation method according to claim 7, characterized in that: During the enzymatic hydrolysis, the enzyme preparation used is a composite enzyme preparation containing trypsin and neutral protease; the amount of the enzyme preparation used is 0.5% of the mass of the second filter residue; And / or, the enzymatic hydrolysis time is 45 min and the temperature is 45°C.

10. The preparation method according to claim 7, characterized in that: During the fermentation, the fermentation bacteria used include Bacillus megaterium, Bacillus licheniformis and Lactococcus lactis; the amount of the fermentation bacteria used is 3% of the mass of the enzymatic residue; And / or, the fermentation time is 14 hours and the temperature is 38°C.

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