A traditional Chinese medicine composition for treating lung cancer and a preparation method thereof

Abstract

The application discloses a traditional Chinese medicine composition for treating lung cancer and a preparation method thereof. The traditional Chinese medicine composition comprises the following raw medicinal materials in parts by weight: Platycladi seeds 5-8 parts, Aescleris stems 19-22 parts, Persicae Nucis 12-15 parts, Radix Pulsatillae 5-8 parts, Coicis Semen 35-38 parts, Benincasae Semen 12-15 parts, Herba Gnaphalii 12-15 parts, Baeckea frutescens 12-15 parts, Citri Reticulatae Pericarpium 5-8 parts and Fructus Jujubae 5-8 parts. The traditional Chinese medicine composition is developed on the basis of the pathogenesis of phlegm and blood stasis of lung cancer in traditional Chinese medicine, combined with classical prescriptions for treating lung diseases in traditional Chinese medicine and laboratory anti-lung cancer drug screening research, and has the effect of inhibiting lung cancer cell growth and good effect on the treatment of lung cancer. Experiments prove that the traditional Chinese medicine composition significantly inhibits lung cancer cell proliferation in vitro and improves the lung cancer pathological process of lung cancer mice in vivo, and is a great innovation of lung cancer drugs.

Description

Technical Field

[0001] This invention relates to a traditional Chinese medicine composition for treating lung cancer and its preparation method, belonging to the field of traditional Chinese medicine technology. Background Technology

[0002] Currently, lung cancer remains the leading cause of cancer incidence and mortality in my country. Radiotherapy, chemotherapy, and immunotherapy are important adjuvant therapies for lung cancer, slowing disease progression and prolonging patient survival. However, these methods have significant side effects, are prone to drug resistance, and reduce patients' quality of life. Furthermore, there is a shortage of clinically available lung cancer medications. Therefore, the search for safe and effective new traditional Chinese medicine treatments for lung cancer has attracted considerable attention from researchers.

[0003] Traditional Chinese medicine (TCM) does not have a specific concept of lung cancer. Based on clinical manifestations and literature, lung cancer symptoms are generally categorized as "cough," "phlegm retention," and "lung accumulation," all belonging to the pulmonary system. TCM considers the lungs to be the receptacle for phlegm and responsible for its dispersing and descending functions. When lung function is impaired, the body's fluids cannot be distributed by the lung qi, leading to the accumulation of phlegm. This phlegm accumulation in the lungs obstructs lung qi, causing blood stasis. The phlegm and blood stasis combine and accumulate in the chest, forming a mass, which is why TCM states that "most lumps in the body are due to phlegm." Therefore, the inventor believes that the mutual binding of phlegm and blood stasis is a core pathogenesis in the development of lung cancer. However, there is currently little research and development of TCM compositions specifically targeting the phlegm-blood stasis pathogenesis of lung cancer. Furthermore, some existing formulations simply combine ancient formulas without achieving the desired therapeutic effect, or their complexity leads to high costs. Therefore, strengthening the research and development of novel TCM compositions for lung cancer is of great significance for improving the clinical prevention and treatment of lung cancer in TCM and enhancing patients' quality of life. Summary of the Invention

[0004] In view of the shortcomings of the prior art, the purpose of this invention is to provide a traditional Chinese medicine composition for treating lung cancer and its preparation method, thereby improving the quality of life of patients, prolonging survival time, and reducing patient mortality in the process of preventing and treating lung cancer.

[0005] To achieve the above objectives, one aspect of the technical solution of the present invention is to provide a traditional Chinese medicine composition for treating lung cancer, comprising the following raw materials in parts by weight: 5-8 parts of Lepidium apetalum, 19-22 parts of Phragmites communis stem, 12-15 parts of Prunus persica kernel, 5-8 parts of Aster tataricus, 35-38 parts of Coix lacryma-jobi, 12-15 parts of Benincasa hispida seed, 12-15 parts of Hedyotis diffusa, 12-15 parts of Selaginella tamariscina, 5-8 parts of Citrus reticulata peel, and 5-8 parts of Ziziphus jujuba.

[0006] Preferably, the traditional Chinese medicine composition for treating lung cancer includes the following raw materials in parts by weight: 8 parts of Lepidium apetalum, 22 parts of Phragmites communis stem, 15 parts of Prunus persica kernel, 8 parts of Aster tataricus, 38 parts of Coix lacryma-jobi, 15 parts of Benincasa hispida seed, 15 parts of Hedyotis diffusa, 15 parts of Selaginella tamariscina, 8 parts of Citrus reticulata peel, and 8 parts of Ziziphus jujuba.

[0007] On the other hand, the technical solution of the present invention is to provide a method for preparing the aforementioned traditional Chinese medicine composition, comprising the following steps:

[0008] (1) Grind the following ingredients into powder: Lepidium seed, reed stem, peach kernel, Aster tataricus, Coix seed, winter melon seed, Hedyotis diffusa, Selaginella tamariscina, and dried tangerine peel; cut the jujube into small pieces and mix them together.

[0009] (2) After mixing, soak in ethanol solution and extract by ultrasonication. After filtering the extract, remove the ethanol from the filtrate using a rotary evaporator, freeze dry, and pulverize into powder to obtain the final product.

[0010] Furthermore, in step (2), the volume concentration of the ethanol solution is 60-80%, and the amount used each time is 8-12 times the total weight of the raw material.

[0011] Furthermore, in step (2), the soaking temperature is room temperature and the soaking time is 20-40 minutes.

[0012] Furthermore, in step (2), ultrasonic extraction is performed multiple times, each time for 60-120 minutes.

[0013] Furthermore, the evaporation temperature of the rotary evaporator in step (2) is 45-55℃.

[0014] On the other hand, the technical solution of the present invention is to provide a method for preparing the aforementioned traditional Chinese medicine composition. The method involves soaking reed stems in pure water, decocting and filtering to obtain a reed stem decoction. The reed stem decoction is then used to soak lepidium seeds, peach kernels, aster, coix seeds, winter melon seeds, *Sedum aizoon*, *Selaginella tamariscina*, dried tangerine peel, and jujubes, followed by decoction. After decoction, the mixture is filtered, and the filtrate is concentrated using a rotary evaporator, then freeze-dried and pulverized into powder to obtain the final product.

[0015] Furthermore, the volume of pure water used is 8-12 times the total weight of the raw materials; the soaking temperature is room temperature, and the soaking time is 20-40 minutes; the decoction is performed multiple times, each time for 45-120 minutes, and the rotary evaporation temperature is 45-55℃.

[0016] On the other hand, the technical solution of the present invention is to provide the application of the aforementioned traditional Chinese medicine composition in the preparation of a drug for treating lung cancer.

[0017] The pharmacological effects of each active pharmaceutical ingredient in this invention are as follows:

[0018] Lepidium seed: It is pungent and bitter in taste, cold in nature, and enters the lung and bladder meridians; it breaks up hard masses and expels pathogens, drains lung heat and promotes water metabolism, and eliminates phlegm and relieves asthma.

[0019] Reed stem: sweet in taste, cold in nature, non-toxic, enters the heart and lung meridians; clears the lungs and detoxifies, stops cough and drains pus.

[0020] Peach kernel: It tastes bitter and sweet, is neutral in nature, and enters the heart, liver, and large intestine meridians; it relieves cough and asthma, invigorates blood and removes blood stasis, and removes blood stasis without harming the body's vital energy.

[0021] Aster: It has a pungent and bitter taste, is warm in nature, and enters the lung meridian. It enters the qi and blood aspects of the lung meridian to regulate the qi and blood of the lung meridian and treat hematemesis, hemoptysis, and hemoptysis.

[0022] Job's tears: sweet and bland in taste, slightly cool in nature, and enters the spleen, stomach, and lung meridians; it has a mild and gentle effect, clearing away damp heat, relieving numbness and draining pus, and promoting urination without harming the body's vital energy.

[0023] Winter melon seeds: sweet in taste, slightly cold in nature, and enter the lung and large intestine meridians; clears the lungs and resolves phlegm, eliminates carbuncles and drains pus, and promotes diuresis.

[0024] Stone-like scutellaria: It has a pungent and bitter taste, is slightly cold in nature, and enters the spleen meridian; it removes blood stasis, disperses nodules, relieves pain, reduces swelling, resolves phlegm, and detoxifies.

[0025] Sago palm: sweet in taste, neutral in nature, enters the lung and liver meridians; clears heat and detoxifies.

[0026] Dried tangerine peel: bitter and pungent in taste, warm in nature, and enters the lung and spleen meridians; it regulates qi, strengthens the spleen, dries dampness, and resolves phlegm.

[0027] Jujube: sweet and neutral in nature, enters the spleen and stomach meridians; it tonifies the spleen and stomach, replenishes qi and generates fluids, regulates the body's defenses, and detoxifies.

[0028] Phlegm and blood stasis are the core pathogenesis of lung cancer, which leads to common symptoms such as cough, phlegm, dry mouth, chest pain, and fatigue. Among them, the dysfunction of lung qi in dispersing and descending manifests as cough, the obstruction of qi flow in the meridians and the abnormal distribution of body fluids result in phlegm and dry mouth. Poor qi flow leads to blood stasis, which manifests as chest pain and fatigue. Phlegm and blood stasis combine and accumulate in the lungs, forming a mass, which eventually leads to disease. Therefore, lung cancer can be treated from the perspective of phlegm and blood stasis. This invention's herbal composition is a fusion and adjustment of the ancient and classical formulas of Weijing Decoction and Tingli Dazao Decoction, making Tinglizi, Lujing, Taoren, and Ziwan the principal herbs. Among them, Tinglizi and Lujing break up hard masses and expel pathogens, drain lung heat and promote urination, resolve phlegm and relieve asthma, stop cough and drain pus. At the same time, Taoren and Ziwan are used to dredge the lung meridian and promote blood circulation and remove blood stasis. The four herbs work together to achieve the effects of resolving phlegm, removing blood stasis and relieving pain. Yiyiren is mild and nourishing, and Dongguaren promotes urination. Shijianchuan removes blood stasis and dissipates nodules, and Shishangbai clears heat and detoxifies. Together they serve as assistant herbs to enhance the effects of the principal herbs in resolving phlegm, removing blood stasis and relieving pain. Chenpi is warm and nourishing, regulates qi and strengthens the spleen and lungs, and assists the principal and assistant herbs in resolving phlegm and removing blood stasis as an adjuvant herb. Dazao detoxifies the herbs and acts as a harmonizing herb, mitigating the cold nature of the herbs. Together, the formula works to resolve phlegm, remove blood stasis and relieve pain.

[0029] Beneficial effects

[0030] This invention relates to a traditional Chinese medicine composition developed based on the pathogenesis of lung cancer in traditional Chinese medicine (TCM) involving phlegm and blood stasis, combined with classic and ancient TCM formulas for treating lung diseases and laboratory screening studies of anti-lung cancer drugs. It exhibits the effect of inhibiting lung cancer cell growth and demonstrates good efficacy in the treatment of lung cancer. Experiments have shown that this TCM composition significantly inhibits lung cancer cell proliferation in vitro and improves the pathological process of lung cancer in mice in vivo, representing a major innovation in lung cancer treatment. Attached Figure Description

[0031] Figure 1 The effect of a traditional Chinese medicine composition on the epigenetic state of lung cancer cells.

[0032] Figure 2 The inhibitory rate of the traditional Chinese medicine composition on the proliferation of lung cancer cells. Detailed Implementation

[0033] The specific embodiments of the present invention will be further described in detail below with reference to examples.

[0034] Example 1

[0035] A traditional Chinese medicine composition for treating lung cancer includes the following raw materials in parts by weight: 5 parts of Lepidium apetalum, 19 parts of Phragmites communis stem, 12 parts of Prunus persica kernel, 5 parts of Aster tataricus, 35 parts of Coix lacryma-jobi, 12 parts of Benincasa hispida seed, 12 parts of Hedyotis diffusa, 12 parts of Selaginella tamariscina, 5 parts of Citrus reticulata peel, and 5 parts of Ziziphus jujuba.

[0036] Its preparation method is as follows:

[0037] (1) Grind the following ingredients into powder: Lepidium seed, reed stem, peach kernel, Aster tataricus, Coix seed, winter melon seed, Hedyotis diffusa, Selaginella tamariscina, and dried tangerine peel. Cut the jujube into small pieces and place them in a beaker.

[0038] (2) Add 8 times the weight of the above raw material and 60% ethanol solution, stir thoroughly, cover the mouth of the beaker with plastic wrap, and soak at room temperature for 20 minutes; then use an ultrasonic extractor to extract the drug solution by stirring with a glass rod every 30 minutes for 60 minutes; after the ultrasonic extraction is completed, cover the mouth of the beaker with nine layers of gauze, pour out the extract, and filter it by vacuum filtration, retaining the filtrate; collect the remaining filter residue and add 8 times the weight of the raw material and 60% ethanol solution for a second ultrasonic extraction, filter, and combine the two filtrates; use a rotary evaporator to evaporate the ethanol in the filtrate at 45°C to obtain a concentrated extract, and finally dry the concentrated extract into a solid state by conventional freeze drying method, and pulverize it into powder, which is the final product, and store at -20°C.

[0039] Example 2

[0040] A traditional Chinese medicine composition for treating lung cancer includes the following raw materials in parts by weight: 6 parts of Lepidium apetalum, 20 parts of Phragmites communis stem, 13 parts of Prunus persica kernel, 6 parts of Aster tataricus, 36 parts of Coix lacryma-jobi, 13 parts of Benincasa hispida seed, 13 parts of Hedyotis diffusa, 13 parts of Selaginella tamariscina, 6 parts of Citrus reticulata peel, and 6 parts of Ziziphus jujuba.

[0041] Its preparation method is as follows:

[0042] (1) Grind the following ingredients into powder: Lepidium seed, reed stem, peach kernel, Aster tataricus, Coix seed, winter melon seed, Hedyotis diffusa, Selaginella tamariscina, and dried tangerine peel. Cut the jujube into small pieces and place them in a beaker.

[0043] (2) Add 10 times the weight of the above raw material and 70% ethanol solution, stir thoroughly, cover the mouth of the beaker with plastic wrap, and soak at room temperature for 30 minutes; then use an ultrasonic extractor to extract the drug solution by stirring with a glass rod every 30 minutes for 90 minutes; after the ultrasonic extraction is completed, cover the mouth of the beaker with nine layers of gauze, pour out the extract, and filter it by vacuum filtration, retaining the filtrate; collect the remaining filter residue and add 10 times the weight of the raw material and 70% ethanol solution for a second ultrasonic extraction, filter, and combine the two filtrates; use a rotary evaporator to evaporate the ethanol in the filtrate at 50°C to obtain a concentrated extract, and finally dry the concentrated extract into a solid state by conventional freeze drying method, and pulverize it into powder, which is the final product, and store at -20°C.

[0044] Example 3

[0045] A traditional Chinese medicine composition for treating lung cancer includes the following raw materials in parts by weight: 7 parts of Lepidium apetalum, 21 parts of Phragmites communis stem, 14 parts of Prunus persica kernel, 7 parts of Aster tataricus, 37 parts of Coix lacryma-jobi, 14 parts of Benincasa hispida seed, 14 parts of Hedyotis diffusa, 14 parts of Selaginella tamariscina, 7 parts of Citrus reticulata peel, and 7 parts of Ziziphus jujuba.

[0046] Its preparation method is as follows:

[0047] (1) Grind the following ingredients into powder: Lepidium seed, reed stem, peach kernel, Aster tataricus, Coix seed, winter melon seed, Hedyotis diffusa, Selaginella tamariscina, and dried tangerine peel. Cut the jujube into small pieces and place them in a beaker.

[0048] (2) Add 12 times the weight of the above raw material and 80% ethanol solution, stir thoroughly, cover the mouth of the beaker with plastic wrap, and soak at room temperature for 40 minutes; then use an ultrasonic extractor to extract the drug solution by stirring with a glass rod every 30 minutes for 120 minutes; after the ultrasonic extraction is completed, cover the mouth of the beaker with nine layers of gauze, pour out the extract, and filter it by vacuum filtration, retaining the filtrate; collect the remaining filter residue and add 12 times the weight of the raw material and 80% ethanol solution for a second ultrasonic extraction, filter, and combine the two filtrates; use a rotary evaporator to evaporate the ethanol in the filtrate at 55°C to obtain a concentrated extract, and finally dry the concentrated extract into a solid state by conventional freeze drying method, and pulverize it into powder, which is the final product, and store at -20°C.

[0049] Example 4

[0050] A traditional Chinese medicine composition for treating lung cancer includes the following raw materials in parts by weight: 8 parts of Lepidium apetalum, 22 parts of Phragmites communis stem, 15 parts of Prunus persica kernel, 8 parts of Aster tataricus, 38 parts of Coix lacryma-jobi, 15 parts of Benincasa hispida seed, 15 parts of Hedyotis diffusa, 15 parts of Selaginella tamariscina, 8 parts of Citrus reticulata peel, and 8 parts of Ziziphus jujuba.

[0051] Its preparation method is as follows:

[0052] (1) Grind the following ingredients into powder: Lepidium seed, reed stem, peach kernel, Aster tataricus, winter melon seed, Job's tears, Hedyotis diffusa, Selaginella tamariscina, and dried tangerine peel. Cut the jujube into small pieces and place them in a beaker.

[0053] (2) Add 10 times the weight of the above raw material and 70% ethanol solution, stir thoroughly, cover the mouth of the beaker with plastic wrap, and soak at room temperature for 30 minutes; then use an ultrasonic extractor to extract the drug solution by stirring with a glass rod every 30 minutes for 90 minutes; after the ultrasonic extraction is completed, cover the mouth of the beaker with nine layers of gauze, pour out the extract, and filter it by vacuum filtration, retaining the filtrate; collect the remaining filter residue and add 10 times the weight of the raw material and 70% ethanol solution for a second ultrasonic extraction, filter, and combine the two filtrates; use a rotary evaporator to evaporate the ethanol in the filtrate at 50°C to obtain a concentrated extract, and finally dry the concentrated extract into a solid state by conventional freeze drying method, and pulverize it into powder, which is the final product, and store at -20°C.

[0054] Example 5

[0055] A traditional Chinese medicine composition for treating lung cancer includes the following raw materials in parts by weight: 8 parts of Lepidium apetalum, 22 parts of Phragmites communis stem, 15 parts of Prunus persica kernel, 8 parts of Aster tataricus, 38 parts of Coix lacryma-jobi, 15 parts of Benincasa hispida seed, 15 parts of Hedyotis diffusa, 15 parts of Selaginella tamariscina, 8 parts of Citrus reticulata peel, and 8 parts of Ziziphus jujuba.

[0056] Its preparation method is as follows:

[0057] (1) Soak the reed stems in pure water at 10 times their weight of the above raw materials, stir thoroughly, cover the mouth of the beaker with plastic wrap, soak at room temperature for 30 minutes, decoct for 45 minutes, filter after decoction to obtain reed stem decoction, add pure water for a second decoction of reed stems, and combine the two decoctions.

[0058] (2) Soak the following ingredients in the decoction of reed stems for 30 minutes: Lepidium seed, peach kernel, Aster tataricus, Coix seed, winter melon seed, Hedyotis diffusa, Selaginella tamariscina, dried tangerine peel, and jujube. Continue to decoct for 45 minutes. After decoction, filter and then decoct a second time. Combine the two decoctions and use a rotary evaporator at 55°C to evaporate the excess water in the decoction to obtain a concentrated extract. Finally, dry the concentrated extract into a solid state using conventional freeze-drying method and pulverize it into powder. Store at -20°C.

[0059] Compare with Example 1

[0060] A control group traditional Chinese medicine composition includes the following raw materials in parts by weight: 22 parts of reed stem, 38 parts of coix seed, 15 parts of peach kernel, and 15 parts of winter melon seed.

[0061] Its preparation method is as follows:

[0062] (1) Grind the reed stems, coix seeds, peach kernels, and winter melon seeds into powder and place them in a beaker;

[0063] (2) Add 10 times the weight of the above raw material and 70% ethanol solution, stir thoroughly, cover the mouth of the beaker with plastic wrap, and soak at room temperature for 30 minutes; then use an ultrasonic extractor to extract the drug solution by stirring with a glass rod every 30 minutes for 90 minutes; after the ultrasonic extraction is completed, cover the mouth of the beaker with nine layers of gauze, pour out the extract, and filter it by vacuum filtration, retaining the filtrate; collect the remaining filter residue and add 10 times the weight of the raw material and 70% ethanol solution for a second ultrasonic extraction, filter, and combine the two filtrates; use a rotary evaporator to evaporate the ethanol in the filtrate at 50°C to obtain a concentrated extract, and finally dry the concentrated extract into a solid state by conventional freeze drying method, and pulverize it into powder, which is the final product, and store at -20°C.

[0064] Compare with Example 2

[0065] A control group traditional Chinese medicine composition includes the following raw materials in parts by weight: 8 parts of Lepidium apetalum and 8 parts of jujube.

[0066] Its preparation method is as follows:

[0067] (1) Grind the lepidium seeds into powder, cut the jujubes into small pieces, and place them in a beaker;

[0068] (2) Add 10 times the weight of the above raw material and 70% ethanol solution, stir thoroughly, cover the mouth of the beaker with plastic wrap, and soak at room temperature for 30 minutes; then use an ultrasonic extractor to extract the drug solution by stirring with a glass rod every 30 minutes for 90 minutes; after the ultrasonic extraction is completed, cover the mouth of the beaker with nine layers of gauze, pour out the extract, and filter it by vacuum filtration, retaining the filtrate; collect the remaining filter residue and add 10 times the weight of the raw material and 70% ethanol solution for a second ultrasonic extraction, filter, and combine the two filtrates; use a rotary evaporator to evaporate the ethanol in the filtrate at 50°C to obtain a concentrated extract, and finally dry the concentrated extract into a solid state by conventional freeze drying method, and pulverize it into powder, which is the final product, and store at -20°C.

[0069] Compare with Example 3

[0070] A control group of traditional Chinese medicine composition includes the following raw materials in parts by weight: 22 parts of reed stem, 38 parts of coix seed, 15 parts of peach kernel, 15 parts of winter melon seed, 8 parts of lepidium seed, and 8 parts of jujube.

[0071] Its preparation method is as follows:

[0072] (1) Grind the reed stems, coix seeds, peach kernels, winter melon seeds, and lepidium seeds into powder, cut the jujubes into small pieces, and place them in a beaker;

[0073] (2) Add 10 times the weight of the above raw material and 70% ethanol solution, stir thoroughly, cover the mouth of the beaker with plastic wrap, and soak at room temperature for 30 minutes; then use an ultrasonic extractor to extract the drug solution by stirring with a glass rod every 30 minutes for 90 minutes; after the ultrasonic extraction is completed, cover the mouth of the beaker with nine layers of gauze, pour out the extract, and filter it by vacuum filtration, retaining the filtrate; collect the remaining filter residue and add 10 times the weight of the raw material and 70% ethanol solution for a second ultrasonic extraction, filter, and combine the two filtrates; use a rotary evaporator to evaporate the ethanol in the filtrate at 50°C to obtain a concentrated extract, and finally dry the concentrated extract into a solid state by conventional freeze drying method, and pulverize it into powder, which is the final product, and store at -20°C.

[0074] Compare with Example 4

[0075] A control group of traditional Chinese medicine composition includes the following raw materials in parts by weight: 22 parts of reed stem, 38 parts of coix seed, 15 parts of peach kernel, 15 parts of winter melon seed, 8 parts of lepidium seed, 8 parts of jujube, 8 parts of gromwell root, 15 parts of dayflower, 15 parts of peony bark, and 8 parts of nard.

[0076] Its preparation method is as follows:

[0077] (1) Grind the reed stems, coix seeds, peach kernels, winter melon seeds, lepidium seeds, gromwell root, dayflower, peony bark, and nard into powder, cut the jujubes into small pieces, and place them in a beaker;

[0078] (2) Add 10 times the weight of the above raw material and 70% ethanol solution, stir thoroughly, cover the mouth of the beaker with plastic wrap, and soak at room temperature for 30 minutes; then use an ultrasonic extractor to extract the drug solution by stirring with a glass rod every 30 minutes for 90 minutes; after the ultrasonic extraction is completed, cover the mouth of the beaker with nine layers of gauze, pour out the extract, and filter it by vacuum filtration, retaining the filtrate; collect the remaining filter residue and add 10 times the weight of the raw material and 70% ethanol solution for a second ultrasonic extraction, filter, and combine the two filtrates; use a rotary evaporator to evaporate the ethanol in the filtrate at 50°C to obtain a concentrated extract, and finally dry the concentrated extract into a solid state by conventional freeze drying method, and pulverize it into powder, which is the final product, and store at -20°C.

[0079] 1. Materials

[0080] 1.1 Cell lines

[0081] Human lung adenocarcinoma epithelial cell line A549 and mouse Lewis lung cancer cells LL / 2 (LLC cells) were both purchased from the Cell Bank of the Chinese Academy of Medical Sciences.

[0082] 1.2 Animals

[0083] C57BL / 6 mice aged 4-6 weeks were purchased from Spiford (Beijing) Biotechnology Co., Ltd.

[0084] 2 methods

[0085] 2.1 Cell Culture

[0086] A549 cells were grown in DMEM complete medium containing 10% (v / v) fetal bovine serum and cultured in a 37°C, 5% CO2 cell culture incubator, with cell passages every 2-3 days.

[0087] 2.2 Mouse Feeding

[0088] The SPF-grade constant-temperature animal room maintains a room temperature of 20-22°C and a relative humidity of 50-70%. In addition, mice are subjected to a 12-hour light and 12-hour dark cycle treatment, and their feed, bedding, and drinking water are changed every 3 days. Their diet is unrestricted.

[0089] 2.3 Observation of the epigenetic status of A549 cells in the normal group and the traditional Chinese medicine combination

[0090] When A549 cells covered more than 80% of the culture dish, the cells were digested with trypsin, the cell pellet was collected, and then resuspended in DMEM complete medium at a concentration of 2.5 × 10⁶ cells / mL. 5 Cells were seeded at a density of 2 mL / well in 6-well plates. After incubation at 37°C and 5% CO2 for 24 h, the culture medium in the wells was discarded, and 2 mL of fresh DMEM complete medium was added to each well. Four control groups were set up: a blank group, an experimental group (the herbal composition from Example 4), a control group 1 (Control Example 1), a control group 2 (Control Example 2), a control group 3 (Control Example 3), and a control group 4 (Control Example 4). The blank group received 20 μL of DMEM complete medium directly. The experimental groups were screened according to the IC50 selection criteria in section 2.4. 30 (The concentration of the traditional Chinese medicine composition when the cell growth inhibition rate reaches 30%), add 10.46 mg / mL of the traditional Chinese medicine composition of the present invention (using DMEM complete medium as solvent), 20 μL / well, so that the working concentration of the traditional Chinese medicine composition of the present invention is 104.6 μg / mL. Control groups 1-4 are respectively added with DMEM complete medium containing the traditional Chinese medicine composition of control examples 1-4, and the dosage is the same as that of the experimental group. Continue to culture at 37℃ and 5% CO2 incubator for 48 h, observe under an inverted microscope and take pictures for recording.

[0091] 2.4 Dose-response relationship of the inhibitory effect of traditional Chinese medicine composition on A549 cell growth as determined by CCK-8 kit

[0092] A549 cells were seeded at a density of 3000 cells / 100 μL / well in 96-well cell culture plates and cultured at 37°C and 5% CO2 for 24 h. At this point, the cells were uniformly dispersed, completely adhered to the plate, and in good condition. The culture medium in the wells was discarded and 100 μL of fresh DMEM complete medium was added. Five groups were set up, including a blank group and a traditional Chinese medicine composition group, with six replicates in each group. The blank group was given 10 μL of DMEM complete medium, and the traditional Chinese medicine composition group was given 10 μL of a solution with concentrations of 312.5, 625, 1250, 2500, and 5000 μg / mL (the traditional Chinese medicine composition in Example 4, with DMEM complete medium as the solvent), so that the working concentrations of the traditional Chinese medicine composition were 31.25, 62.5, 125, 250, and 500 μg / mL, respectively. After incubation at 37℃ and 5% CO2 for 48 hours, the culture medium was removed, and 100 μL of DMEM complete medium containing 10% (v / v) CCK-8 kit solution was added to each well. After incubation at 37℃ and 5% CO2 for 30 minutes, the absorbance (OD value) of each well was immediately measured using a microplate reader at 450 nm. The absorbance was measured according to the formula:

[0093] Inhibition rate (%) = (1 - Average A of the pores in the traditional Chinese medicine composition) 450值 / Average A of blank holes 450值 )×100%

[0094] Calculate the IC of traditional Chinese medicine composition 30 Value: Repeat the above experiment three times, import the corresponding inhibition rates into an Excel spreadsheet, insert a scatter plot to generate a trend line, and calculate the IC50 of the herbal composition based on the formula corresponding to the trend line. 30 value.

[0095] 2.5 Effects of Traditional Chinese Medicine Composition on LLC Subcutaneous Xenograft Mice

[0096] LLC cells in the logarithmic growth phase were washed with PBS (phosphate-buffered saline), digested with trypsin, centrifuged, and the pellet was precipitated. Finally, the cell pellet was resuspended in PBS to a concentration of 1×10⁻⁶. 7LLC cell suspension was prepared by subcutaneous injection of 150 μL PBS buffer (blank group) into the right anterior axilla of 6 mice using a microsyringe, and 150 μL LLC cell suspension into the right anterior axilla of the remaining mice to construct subcutaneous xenograft mice for lung cancer. Successful modeling was indicated by the presence or palpation of a regular tumor the size of a mung bean in the right axilla of the modeling mice after 7 days. The modeling mice were divided into a model group, a traditional Chinese medicine composition experimental group (the traditional Chinese medicine composition in Example 4), a traditional Chinese medicine composition control group 1 (control example 1), a traditional Chinese medicine composition control group 2 (control example 2), a traditional Chinese medicine composition control group 3 (control example 3), a traditional Chinese medicine composition control group 4 (control example 4), and a cisplatin group, with 6 mice in each group. Mice in the traditional Chinese medicine composition experimental group and the traditional Chinese medicine composition control group were administered cisplatin by gavage once daily, while mice in the cisplatin group were administered cisplatin via intraperitoneal injection once every other day. Dosage calculation: Based on the body surface area method, the equivalent dose of the crude Chinese herbal medicine in mice was calculated to be 19.5 g / kg, corresponding to a dosage of 6.5 g / kg of the Chinese herbal medicine powder composition, with a dosage volume of 0.1 mL / 10 g (the dosage was the same for experimental and control groups 1-4). The cisplatin group received an intraperitoneal injection of 1 mg / kg cisplatin (using sterile saline as the solvent), 0.1 mL / 10 g. The model group and the blank group received an equal volume of saline via gavage. Tumor volume in the axilla of mice was measured every two days. After 14 days of treatment, mice were euthanized by cervical dislocation, the tumor was removed, and the final tumor volume was measured, and the tumor inhibition rate was calculated.

[0097] Mouse tumor volume calculation: The longest diameter L (mm) and the maximum transverse diameter W (mm) of the subcutaneous tumor in each mouse were measured using electronic vernier calipers. The tumor volume V (mm²) of each group of mice was calculated using the Lee's method. 3 )=L×W 2 / 2.

[0098] Drug tumor inhibition rate E: E = 1 - average tumor volume of mice in the drug treatment group / average tumor volume of mice in the model group.

[0099] Drug tumor inhibition percentage: Tumor inhibition percentage = E × 100%.

[0100] To evaluate whether the combination therapy has a synergistic effect on tumor suppression: Q = E 实验组 / 对照例3 / 对照例4 / (E 对照例1 +E 对照例2 -E 对照例1 *E 对照例2 The expression is given by the expression, where Q < 0.85 indicates that the two are antagonistic, 0.85 ≤ Q < 1.15 indicates that the two are additive, and Q ≥ 1.15 indicates that the two are synergistic.

[0101] 2.6 Effects of Traditional Chinese Medicine Composition on Body Weight in LLC Subcutaneous Xenograft Mice

[0102] After administration, the mice were observed every two days for their diet, activity level, hair sparseness and luster, perception of the external environment, and other general conditions, and were weighed using a small animal electronic scale.

[0103] 2.7 Statistical Methods

[0104] One-way ANOVA was performed on the measurement data using GraphPad Prism 9.5.0 software. The experimental data results are expressed as mean values.

[0105] 3 Experimental Results

[0106] 3.1 Effects of Traditional Chinese Medicine Composition on the Growth and Morphology of A549 Cells

[0107] Under an inverted microscope, cells in the blank control group adhered to the microstructure, exhibiting polygonal or spindle-shaped growth, smooth surfaces, clear cell boundaries, and a small number of cells possessing pseudopodia. Compared to the blank control group, some cells in control group 2 showed granular material, but the overall cell state remained largely unchanged. Some cells in control group 1 also showed granular material and a small amount of necrotic blank areas. Both control groups 3 and 4 showed granular material and necrotic blank areas in some cells. However, the cells in the herbal composition group (experimental group) of this invention had extremely coarse cytoplasm, filled with a large amount of granular material, and exhibited extensive necrotic blank areas within the cells, suggesting that treatment with the herbal composition of this invention worsened the condition of tumor cells, even leading to large-scale cell death (see...). Figure 1 This indicates that the tumor cell killing effects of Control Examples 3 and 4 are higher than those of Control Examples 1 and 2, but the efficacy of the traditional Chinese medicine composition in this invention is significantly higher than that of the traditional Chinese medicine compositions in Control Examples 3 and 4.

[0108] 3.2 Effects of Traditional Chinese Medicine Composition on A549 Cell Proliferation

[0109] The traditional Chinese medicine composition can inhibit the proliferation of A549 lung cancer cells, and the inhibitory effect on lung cancer cell proliferation shows a significant dose-response relationship with increasing concentration of the traditional Chinese medicine composition (see Table 1 and Table 2). Figure 2 Simultaneously, based on the dose-response curve, the IC was calculated. 30 The value was 104.6 μg / mL.

[0110] Table 1: Relationship between the growth inhibition rate of traditional Chinese medicine composition on A549 lung cancer cells and drug concentration (unit: μg / mL)

[0111]

[0112] 3.3 Inhibitory effect of traditional Chinese medicine composition on tumor growth in LLC subcutaneous xenograft mice

[0113] Eight days after subcutaneous injection of LLC cell suspension, all model mice developed tumors the size of soybeans on the right subcutaneous surface. At the end of the experiment, compared with the model group, the cisplatin group showed a significant reduction in tumor volume, indicating that cisplatin inhibited tumor growth in the mice with extremely significant inhibitory effects. Compared with the model group, control groups 1 and 2 showed smaller tumor volumes, but the difference was not statistically significant, suggesting that control groups 1 and 2 had weak inhibitory effects on tumor growth, while control groups 3 and 4 showed significant reductions in tumor volume. Compared with the model group, the experimental group of mice with the traditional Chinese medicine composition showed a significant reduction in tumor volume, indicating that the traditional Chinese medicine composition of this invention can significantly inhibit tumor growth in mice, and the inhibitory effect is significant. Compared with the cisplatin group, the experimental group of mice with the traditional Chinese medicine composition showed an increase in tumor volume, suggesting that the inhibitory effect of the traditional Chinese medicine composition of this invention on tumor growth in mice is slightly weaker than that of cisplatin, but the difference was not statistically significant. Compared with the experimental group of the traditional Chinese medicine composition, control groups 1, 2, 3, and 4 showed significant inhibitory effects on tumor growth. The effects were weaker and significantly different. Furthermore, compared to control group 1 and control group 2 mice, control group 3 and control group 4 showed stronger tumor growth inhibition than control group 1 and control group 2. However, based on the Q-value method for evaluating the synergistic effect of combined drug administration on tumor inhibition, the Q values ​​for control examples 3 and 4 were 1.047 and 1.087, respectively, indicating that the tumor growth inhibition effects of control examples 3 and 4 were merely the sum of the effects of control examples 1 and 2, without significant synergistic effects. Meanwhile, compared to control group 1 and control group 2 mice, the tumor volume in the experimental group of mice with the traditional Chinese medicine composition was significantly reduced, and the Q value was 2.050, suggesting that the tumor inhibition effect of the traditional Chinese medicine composition of this invention is not merely the sum of the effects of control examples 1 and 2, but rather exhibits a highly significant synergistic effect (see Table 2).

[0114] Table 2: Effects of Traditional Chinese Medicine Composition on Tumor Volume in LLC Subcutaneous Xenograft Mice (mm) 3 )

[0115]

[0116] * ^ indicates P < 0.01 compared to the model group; ^ indicates P < 0.05 compared to the cisplatin group. $ This indicates that, compared with the experimental group of traditional Chinese medicine composition, P < 0.01.

[0117] 3.4 Effects of Traditional Chinese Medicine Composition on General Condition and Body Weight of LLC Subcutaneous Xenograft Mice

[0118] No mice died during the experiment. Mice in the control group had shiny, smooth fur, were agile, and had a normal diet. Mice in the model group had sparse fur, listless eyes, decreased activity, and were emaciated, although their overall weight increased significantly due to rapid tumor growth in the later stages. Mice in the cisplatin group had dry, sparse fur, listless eyes, decreased activity, reduced appetite, and were emaciated. Mice in the herbal composition experimental group had smooth fur, were agile, had a normal diet, and normal body shape. Mice in control groups 1, 2, 3, and 4 all exhibited sparse fur, listless eyes, decreased activity, and emaciated body shape (see Table 3). These results indicate that cisplatin significantly reduced the survival status and quality of life of mice with tumors while inhibiting tumor growth. The herbal composition of this invention significantly improved the survival status and emaciated body shape of mice with lung cancer. Controls 1, 2, 3, and 4 had a weaker effect on improving the quality of life and emaciated body shape of mice with lung cancer, and these effects were all weaker than those of the herbal composition of this invention.

[0119] Table 3: Effects of traditional Chinese medicine composition on mean body weight in LLC subcutaneous xenograft mice (N=6)

[0120]

[0121] # This indicates that compared to the control group, P < 0.01; * ^ indicates that compared with the model group, P < 0.01; ^ indicates that compared with the cisplatin group, P < 0.01; $ This indicates that, compared with the experimental group of traditional Chinese medicine composition, P < 0.01.

[0122] 4. Conclusion

[0123] 4.1 The herbal composition of the present invention showed good inhibitory effect on the proliferation of lung cancer cells in in vitro cell experiments, and the effect was dose-dependent.

[0124] 4.2 The herbal composition of the present invention can significantly inhibit the growth of LLC subcutaneous xenograft tumors in mice and can significantly improve the general condition of lung cancer mice, such as lethargy, reduced activity, overall emaciation, and poor appetite.

[0125] 4.3 Based on classical and ancient prescriptions, the combination and adjustment of the traditional Chinese medicine composition of this invention has a significant effect on inhibiting the growth of lung cancer.

[0126] 4.4 The herbal composition of this invention has a good inhibitory effect on lung cancer growth and has few toxic side effects. It is an innovation in the treatment of lung cancer and has important clinical significance.

Claims

1. A traditional Chinese medicine composition for treating lung cancer, characterized in that, It is made from the following raw materials in parts by weight: 5-8 parts of Lepidium seed, 19-22 parts of reed stem, 12-15 parts of peach kernel, 5-8 parts of Aster tataricus, 35-38 parts of Coix seed, 12-15 parts of winter melon seed, 12-15 parts of Hedyotis diffusa, 12-15 parts of Selaginella tamariscina, 5-8 parts of dried tangerine peel, and 5-8 parts of jujube.

2. The traditional Chinese medicine composition for treating lung cancer according to claim 1, characterized in that, It is made from the following raw materials in parts by weight: 8 parts of Lepidium seed, 22 parts of reed stem, 15 parts of peach kernel, 8 parts of Aster tataricus, 38 parts of Coix seed, 15 parts of winter melon seed, 15 parts of Hedyotis diffusa, 15 parts of Selaginella tamariscina, 8 parts of dried tangerine peel, and 8 parts of jujube.

3. A method for preparing the traditional Chinese medicine composition as described in claim 1 or 2, characterized in that, Includes the following steps: (1) Grind the following ingredients into powder: Lepidium seed, reed stem, peach kernel, Aster tataricus, Coix seed, winter melon seed, Hedyotis diffusa, Selaginella tamariscina, and dried tangerine peel; cut the jujube into small pieces and mix them together. (2) After mixing, soak in ethanol solution and extract by ultrasonication. After filtering the extract, remove the ethanol from the filtrate using a rotary evaporator, freeze dry, and pulverize into powder to obtain the final product.

4. The preparation method according to claim 3, characterized in that, In step (2), the volume concentration of the ethanol solution is 60-80%, and the amount used each time is 8-12 times the total weight of the raw material.

5. The preparation method according to claim 3, characterized in that, In step (2), the soaking temperature is room temperature and the soaking time is 20-40 minutes.

6. The preparation method according to claim 3, characterized in that, In step (2), ultrasonic extraction is performed multiple times, each time for 60-120 minutes.

7. The preparation method according to claim 3, characterized in that, In step (2), the evaporation temperature of the rotary evaporator is 45-55℃.

8. A method for preparing the traditional Chinese medicine composition as described in claim 1 or 2, characterized in that, Soaking and decocting reed stems in pure water, followed by filtration, yields a reed stem decoction. The decoction is then used to soak Lepidium apetalum, peach kernel, Aster tataricus, Coix lacryma-jobi, winter melon seed, Hedyotis diffusa, Selaginella tamariscina, dried tangerine peel, and jujubes. After decocting, the decoction is filtered, and the filtrate is concentrated using a rotary evaporator. Finally, the filtrate is freeze-dried and pulverized into powder to obtain the final product.

9. The preparation method according to claim 8, characterized in that, The volume of pure water used should be 8-12 times the total weight of the raw materials; the soaking temperature should be room temperature, and the soaking time should be 20-40 minutes; the decoction should be performed multiple times, each time for 45-120 minutes; the rotary evaporation temperature should be 45-55℃.

10. The use of a traditional Chinese medicine composition as described in claim 1 or 2 in the preparation of a drug for treating lung cancer.

Images