Traditional Chinese medicine composition for treating lung cancer

Abstract

The invention provides a traditional Chinese medicine composition for treating lung cancer, and relates to the technical field of traditional Chinese medicine pharmacy. Specifically, the traditional Chinese medicine composition for treating lung cancer is prepared from radix aconiti preparata, rhizoma pinelliae preparata, radix asteris, rhizoma curcumae, poria cocos, cortex moutan, prepared fructus evodiae, ginseng, pericarpium zanthoxyli and liquorice, and has the effects of warming yang, dispelling cold, reducing phlegm, removing stasis, tonifying qi and strengthening the body resistance. Proliferation and migration of cancer cells can be effectively inhibited, and apoptosis of the cancer cells is promoted; the compound has an obvious treatment effect on tumor model mice, can obviously improve the survival state of the tumor model mice, has an obvious expression inhibition effect on a cell proliferation marker Ki-67, realizes an obvious inhibition effect on tumor growth, and can obviously improve the pathological morphology of tumor tissues. The traditional Chinese medicine composition provided by the invention provides a new direction for the development of medicines for treating lung cancer.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of traditional Chinese medicine, in particular to a traditional Chinese medicine composition for treating lung cancer. BACKGROUND

[0002] Lung cancer has high morbidity and mortality, and poor prognosis, and short survival period. At present, its main treatment methods include surgery, radiotherapy and chemotherapy, targeted therapy and immunotherapy, etc. Due to the limitations of surgical treatment and the emergence of cell toxicity and tolerance caused by radiotherapy and chemotherapy, the prognosis of lung cancer patients is poor; in the past 20 years, with the emergence and development of various targeted therapies, lung cancer has made great progress in research, screening and personalized treatment (precision medicine), which can specifically kill tumor cells, reduce toxicity, prolong the progression-free survival (PFS) of patients, and improve the quality of life, but most of the patients who are treated effectively will face the problem of drug resistance and tumor recurrence or metastasis, thus leading to the bottleneck of lung cancer treatment.

[0003] With the continuous development of modern medical technology, surgery, radiotherapy (hereinafter referred to as radiotherapy), chemotherapy (hereinafter referred to as chemotherapy), targeted therapy, immunotherapy and other treatment methods have greatly improved the prognosis of lung cancer patients, and patients may have bone marrow hematopoietic function inhibition, gastrointestinal reactions, liver, kidney, neurotoxicity, and normal immune function is weakened after traditional treatment, which causes great pain to patients and leads to a decline in quality of life. Traditional Chinese medicine has good effect in the treatment of lung cancer, which can relieve the symptoms of patients after traditional treatment, prolong survival time and improve quality of life, so it is more and more widely used in cancer treatment. SUMMARY

[0004] (I) Technical problems to be solved

[0005] In view of the deficiencies of the prior art, the present application provides a traditional Chinese medicine composition for treating lung cancer, which has the effects of warming yang, dispelling cold, resolving phlegm and removing blood stasis, and tonifying qi and strengthening the body resistance.

[0006] (II) Technical solutions

[0007] In order to achieve the above purpose, the present application is realized by the following technical solutions:

[0008] Firstly, the present application provides a traditional Chinese medicine composition for treating lung cancer, which is made of the following weight parts of traditional Chinese medicine raw materials: 5-20 parts of prepared ephedra, 3-15 parts of radix pinelliae, 3-15 parts of radix scrophulariae, 3-15 parts of radix curcumae, 3-15 parts of poria cocos, 3-15 parts of cortex moutan, 3-15 parts of prepared evodia, 3-15 parts of radix ginseng, 3-15 parts of zanthoxyli, and 3-15 parts of radix glycyrrhizae.

[0009] Further, the traditional Chinese medicine composition is made from the following raw materials by weight: prepared monkshood 10-20 parts, clear pinellia 3-10 parts, common yam 3-10 parts, curcuma zedoaria 3-10 parts, poria 3-10 parts, red peony root 3-10 parts, prepared evodia 3-10 parts, ginseng 5-15 parts, zanthoxylum 3-10 parts, and licorice 3-10 parts.

[0010] Further, the traditional Chinese medicine composition is made from the following raw materials by weight: prepared monkshood 10-20 parts, clear pinellia 3-10 parts, common yam 3-10 parts, curcuma zedoaria 3-10 parts, poria 3-10 parts, red peony root 3-10 parts, prepared evodia 3-10 parts, ginseng 5-15 parts, zanthoxylum 3-10 parts, and licorice 3-10 parts.

[0011] In addition, the traditional Chinese medicine composition provided by the application is used for preparing a drug for treating lung cancer, and the drug is prepared from the traditional Chinese medicine in any one of the oral dosage forms of powder, granules, pills, decoction, powder, oral liquid, tablets, and capsules by adding a medically acceptable excipient.

[0012] Lung cancer belongs to the category of "lung accumulation" and "qi stagnation" in traditional Chinese medicine. The pathogenesis is based on deficiency of vital qi, and accumulation of phlegm turbidity, blood stasis, and cold toxin in the lung. The disease course is complicated by deficiency and excess, and cold and heat. Therefore, the treatment should follow the principle of "regulating cold and heat, and combining attack and tonification". Based on this, the inventors provide a traditional Chinese medicine composition for treating lung cancer, which is composed of prepared monkshood, clear pinellia, common yam, curcuma zedoaria, poria, red peony root, prepared evodia, ginseng, zanthoxylum, and licorice.

[0013] In the prescription, prepared monkshood is pungent and bitter in nature, and hot in property. It is distributed in the heart, liver, kidney, and spleen meridians. After processing, the toxicity of prepared monkshood is reduced, but the warm and unobstructed property remains. The disease course of lung cancer patients is long, and the yang qi is easily deficient. Cold evil is easily condensed, and phlegm turbidity and blood stasis are accumulated in the lung collaterals, resulting in lung failure, qi stagnation, and symptoms such as cough, hemoptysis, chest pain, and shortness of breath. Prepared monkshood can directly break the cold toxin accumulation in the lung with its strong and warm yang qi, and can also warm the meridians and remove stasis, dry dampness and resolve phlegm, and resolve the mutual accumulation of phlegm turbidity and cold evil. Therefore, it is the core of the whole prescription for breaking yin and dispersing cold, dredging collaterals, and removing accumulation. Therefore, it is the monarch herb.

[0014] Evodia rutaecarpa (Juss.) Benth. et Hook. f. var. officinalis Dode is pungent and hot, and its meridian is liver, spleen and stomach. It has the functions of warming middle energizer and dispelling cold, and descending adverse qi and relieving pain. Both of them are warm-heat drugs, and their pungent and hot natures are compatible with the monarch drug Radix Aconiti. The combination of the two drugs can help Radix Aconiti to warm and disperse cold stagnation in the middle and lower energizers, and target the pathogenesis of lung cancer, which is cold evil predominating in the lung and yang qi deficiency. It can further dispel cold evil and warm yang qi, so that the lung collaterals are warmed and blood stasis is easily dispersed, and phlegm and turbid fluid are warmed and easily eliminated. Pinellia ternata (Thunb.) Breit. is pungent and warm, and its meridian is spleen, stomach and lung. It can dry dampness, resolve phlegm, descend adverse qi and relieve vomiting. Allium atrofuscum Regel is bitter and sweet, and its meridian is lung. It is good at moistening the lung, promoting qi and resolving phlegm and relieving cough. Its nature is slightly moistening, and it can resolve phlegm and prevent dryness from damaging the lung when combined with Pinellia ternata (Thunb.) Breit. The combination of the two drugs can resolve phlegm and moisten the lung to prevent dryness from being too excessive to damage the lung. The combination of the two drugs can enhance the resolving phlegm of the monarch drug and take into account the nature of the lung, which likes moistening and dislikes dryness. It can resolve phlegm without damaging the lung, and moisten the lung without assisting phlegm. Evodia rutaecarpa (Juss.) Benth. et Hook. f. var. officinalis Dode, Allium atrofuscum Regel, Pinellia ternata (Thunb.) Breit. and Allium bakeri Regel are all the minister drugs, which can warm yang, disperse cold, resolve phlegm and relieve cough from multiple dimensions to assist the monarch drug, so that the whole prescription can eliminate evil more specifically and completely. At the same time, it takes into account the physiological characteristics of the lung, and is an important assistant in eliminating evil.

[0015] Curcuma zedoaria (Crantz) Hance is pungent, bitter and warm, and its meridian is liver and spleen. It has the functions of breaking blood and promoting qi, and eliminating stagnation and relieving pain. It can break blood and remove stasis, and can be used to treat blood stasis in the lung, so that blood stasis is dispersed and qi movement is unobstructed. Radix Paeoniae Alba is bitter, pungent and cold, and its meridian is heart, liver and kidney. It has the functions of cooling blood and removing stasis, and clearing heat. Its nature is cold, which can help Curcuma zedoaria (Crantz) Hance to remove blood stasis, and can also regulate the warm and dry nature of the monarch drug Radix Aconiti and the minister drugs Evodia rutaecarpa (Juss.) Benth. et Hook. f. var. officinalis Dode and Allium bakeri Regel, so as to prevent the warm and dry nature from being too excessive to consume yin blood. At the same time, it can treat low fever and hemoptysis caused by heat due to long-term stasis in lung cancer patients. Radix et Rhizoma Ginseng is slightly warm, and its meridian is spleen, lung and heart. It can tonify primordial qi, tonify the spleen and lung, and produce fluid and nourish blood. It is a tonic drug, which can be used to treat primordial qi deficiency and spleen and lung qi deficiency in lung cancer patients. It can tonify lung qi to assist lung descending, and tonify spleen qi to assist the spleen as the “postnatal foundation” of the lung, so that the lung qi is sufficient and the spleen qi is strong, which can eliminate evil. Radix Poriae is neutral, and its meridian is heart, spleen and kidney. It can drain water and dampness, and calm the heart. Its nature is neutral, which can help Radix et Rhizoma Ginseng to tonify the spleen and lung, and drain water and dampness to assist Pinellia ternata (Thunb.) Breit. to resolve phlegm, which embodies the principle of “treating phlegm by treating dampness, and phlegm will be eliminated when dampness is eliminated”. It can also calm the heart and relieve restlessness and insomnia in lung cancer patients. The combination of Radix et Rhizoma Ginseng and Radix Poriae can tonify and drain, which can enhance the whole prescription in supporting the healthy and laying the foundation for eliminating evil, and embodies the principle of “supporting the healthy without leaving evil”. Curcuma zedoaria (Crantz) Hance, Radix Paeoniae Alba, Radix et Rhizoma Ginseng and Radix Poriae are all the minister drugs, which can help the monarch and minister drugs to remove blood stasis, resolve phlegm and support the healthy, and regulate the warm and dry nature of the monarch and minister drugs, so that the whole prescription can eliminate evil and support the healthy, and balance the warm and dry nature and the cool and moist nature.

[0016] Finally, licorice is used as the guiding herb. Licorice is sweet and neutral in nature, entering the heart, lung, spleen, and stomach meridians. Its sweet taste can moderate the potent toxicity of the principal herb, Aconitum carmichaelii, as well as the warming and drying properties of the assistant herbs, Evodia rutaecarpa and Zanthoxylum bungeanum. It also moderates the blood-breaking power of the adjuvant herb, Curcuma zedoaria, thus making the overall effect of the formula more balanced and preventing it from being too harsh or damaging to the body's vital energy. On the other hand, licorice can "harmonize the properties of the herbs," preventing warming herbs from being too drying, cooling herbs from being too cold, tonifying herbs from being too stagnant, and purgative herbs from being too drastic. This allows all the herbs in the formula to work synergistically, jointly exerting the effects of warming yang and dispelling cold, resolving phlegm and removing blood stasis, and tonifying qi and supporting the body's vital energy. Furthermore, licorice enters the lung meridian and also has a certain effect of moistening the lungs and relieving cough, which can assist the assistant herbs in alleviating the cough symptoms of lung cancer, thus also having an "adjuvant" effect.

[0017] The complete formula has the following characteristics:

[0018] Extremely pungent and hot, it attacks and expels deep-seated cold: The chief herb, Aconitum carmichaelii, warms the lungs and dispels cold toxins, serving as the core of the entire formula to expel pathogens. The assistant herbs, Evodia rutaecarpa and Zanthoxylum bungeanum, help the chief herb form a powerful warming and cold-dispelling effect.

[0019] Treating both phlegm and blood stasis, addressing both the root cause and the symptoms: not only dispelling cold, but also using the assistant herbs Pinellia ternata and Aster tataricus to help the principal herbs resolve phlegm and enhance the ability to expel pathogens, while the adjuvant herbs Curcuma zedoaria and Paeonia suffruticosa remove blood stasis, unblock the meridians, and counteract the warming and drying effects, thus comprehensively clearing away pathological products.

[0020] The approach combines attack and tonification, addressing both pathogenic factors and the body's resistance: While using strong medicines to attack pathogenic factors, it also heavily utilizes Poria cocos, ginseng, and licorice to invigorate qi, strengthen the spleen, and support the body's resistance, thus attacking pathogenic factors without harming the body's resistance.

[0021] Using both hot and cold herbs to control their properties and preserve their efficacy: Adding peony bark, which is cold in nature, to a large amount of hot herbs helps to control their drying and pungent properties, prevents damage to yin, and makes the overall formula more comprehensive and stable.

[0022] The formula works synergistically to warm the yang and dispel cold, resolve phlegm and remove blood stasis, and invigorate qi and support the body's resistance. It adheres to the principle of regulating both cold and heat and combining attack and tonification, and is mainly used to treat syndromes of mixed cold and heat and phlegm and blood stasis.

[0023] Furthermore, we evaluated and explored the efficacy of the traditional Chinese medicine composition of this invention in the treatment of lung cancer. In vitro, the traditional Chinese medicine composition of this invention effectively inhibited the proliferation and migration of cancer cells and promoted apoptosis. In vivo, the traditional Chinese medicine composition of this invention showed significant therapeutic effects on tumor model mice, significantly improving their survival status, significantly inhibiting the expression of the cell proliferation marker Ki-67, achieving a significant inhibitory effect on tumor growth, and significantly improving the pathological morphology of tumor tissue. The traditional Chinese medicine composition provided by this invention offers a new direction for the development of drugs for the treatment of lung cancer. Attached Figure Description

[0024] Figure 1 The value represents the proliferation rate of A549 lung cancer cells; compared with the blank control group, **P<0.01.

[0025] Figure 2The migration rate of A549 lung cancer cells was statistically significant; compared with the blank control group, **P<0.01.

[0026] Figure 3 The apoptosis rate of A549 lung cancer cells was calculated; compared with the blank control group, **P<0.01; compared with the traditional Chinese medicine group, *P<0.05.

[0027] Figure 4 Tumor weight (A) and volume (BC) of mice in each group are shown; compared with the model control group, *P<0.05, **P<0.01.

[0028] Figure 5 HE staining results of tumor tissues from mice in each group.

[0029] Figure 6 Immunohistochemical staining results of tumor tissues in mice of each group (A) and the proportion of Ki-67 positive tissues in tumor tissues; compared with the model control group, *P<0.05, **P<0.01. Detailed Implementation

[0030] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0031] Example 1

[0032] A traditional Chinese medicine composition for treating lung cancer, the composition being made from the following traditional Chinese medicine raw materials in parts by weight: 5 parts of processed Aconitum carmichaelii, 3 parts of processed Pinellia ternata, 3 parts of Aster tataricus, 3 parts of Curcuma zedoaria, 3 parts of Poria cocos, 3 parts of Paeonia suffruticosa, 3 parts of processed Evodia rutaecarpa, 3 parts of ginseng, 3 parts of Zanthoxylum bungeanum, and 3 parts of Glycyrrhiza uralensis.

[0033] Using the above-mentioned proportions of traditional Chinese medicine as raw materials, and supplemented with medically acceptable excipients, an oral dosage form is prepared, wherein the dosage form is any one of powder, granules, pills, decoction, powder, oral liquid, tablets, and capsules.

[0034] Specifically, the traditional Chinese medicine pills are prepared according to the following steps:

[0035] Weigh each Chinese herbal medicine ingredient according to the above-mentioned weight proportions, wash and dry them, grind them into fine powder, pass them through a 120-mesh sieve, and mix them evenly to obtain the raw Chinese herbal medicine powder. Take honey according to the weight ratio of raw Chinese herbal medicine powder to honey of 1:1.2, heat the honey to boiling, remove foam and impurities, refine it to a temperature of 110-120℃, with a water content of less than 15%, a light yellow color, and moderate viscosity. Then add the honey to the raw Chinese herbal medicine powder while it is still hot, stir quickly to make the honey and raw Chinese herbal medicine powder fully blended to form a medicinal ball. Divide the medicinal ball into small pieces, roll them into strips with a diameter of 5mm, then cut them into small segments of 5mm, roll them into rounds to obtain honey pills, dry them at a temperature below 60℃ until the water content is less than 10%, which is the Chinese herbal medicine pill, 0.5±0.15g / pill.

[0036] Specifically, the traditional Chinese medicine decoction is prepared according to the following steps:

[0037] Weigh each Chinese herbal medicine ingredient according to the above weight proportions, wash them, soak them in 4 times their weight of water for 30 minutes, boil them over high heat for 30 minutes, collect the decoction, repeat the decoction twice, combine the decoctions, evaporate and concentrate to 400 mL, divide into 2 doses, 200 mL each time.

[0038] Specifically, the oral liquid of traditional Chinese medicine is prepared according to the following steps:

[0039] Weigh the above-mentioned proportions of each Chinese herbal medicine raw material, wash, dry, pulverize, and pass through a 10-mesh sieve; then add 10 times the amount of water, soak for 40 minutes, bring to a boil over high heat, then simmer over low heat for 90 minutes, filter, collect the filtrate, add 8 times the amount of water to the residue, bring to a boil over high heat, then simmer over low heat for 60 minutes, filter, combine the two filtrates, and concentrate under reduced pressure to a relative density of 1.10-1.15 to obtain the water extract; then add 95% ethanol to the water extract to achieve an alcohol content of 6%. 0%, let stand for more than 24 hours, take the supernatant, recover the ethanol until there is no alcohol odor, then let stand at 4℃ for more than 48 hours, then centrifuge to remove the precipitate to obtain the alcohol extract; then add 5% sucrose and 2% potassium sorbate by weight of the alcohol extract, stir to dissolve, then add purified water to a total volume of 1000mL, adjust the pH to 5.5-6.5 with citric acid, then filter through 0.45μm micropores, then sterilize, fill into 10mL ampoules, and seal with nitrogen.

[0040] Example 2

[0041] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine composition is made from the following traditional Chinese medicine raw materials in parts by weight: 20 parts of processed Aconitum carmichaelii, 15 parts of processed Pinellia ternata, 15 parts of Aster tataricus, 15 parts of Curcuma zedoaria, 15 parts of Poria cocos, 15 parts of Paeonia suffruticosa, 15 parts of processed Evodia rutaecarpa, 15 parts of ginseng, 15 parts of Zanthoxylum bungeanum, and 15 parts of Glycyrrhiza uralensis.

[0042] Example 3

[0043] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine composition is made from the following traditional Chinese medicine raw materials in parts by weight: 18 parts of processed Aconitum carmichaelii, 10 parts of processed Pinellia ternata, 12 parts of Aster tataricus, 12 parts of Curcuma zedoaria, 10 parts of Poria cocos, 8 parts of Paeonia suffruticosa, 8 parts of processed Evodia rutaecarpa, 10 parts of ginseng, 12 parts of Zanthoxylum bungeanum, and 12 parts of Glycyrrhiza uralensis.

[0044] Example 4

[0045] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 10 parts of processed Aconitum carmichaelii, 3 parts of processed Pinellia ternata, 3 parts of Aster tataricus, 3 parts of Curcuma zedoaria, 3 parts of Poria cocos, 3 parts of Paeonia suffruticosa, 3 parts of processed Evodia rutaecarpa, 5 parts of ginseng, 3 parts of Zanthoxylum bungeanum, and 3 parts of Glycyrrhiza uralensis.

[0046] Example 5

[0047] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 20 parts of processed Aconitum carmichaelii, 10 parts of processed Pinellia ternata, 10 parts of Aster tataricus, 10 parts of Curcuma zedoaria, 10 parts of Poria cocos, 10 parts of Paeonia suffruticosa, 10 parts of processed Evodia rutaecarpa, 15 parts of ginseng, 10 parts of Zanthoxylum bungeanum, and 10 parts of Glycyrrhiza uralensis.

[0048] Example 6

[0049] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 12 parts of processed Aconitum carmichaelii, 4 parts of processed Pinellia ternata, 4 parts of Aster tataricus, 4 parts of Curcuma zedoaria, 4 parts of Poria cocos, 4 parts of Paeonia suffruticosa, 4 parts of processed Evodia rutaecarpa, 6 parts of ginseng, 4 parts of Zanthoxylum bungeanum, and 4 parts of Glycyrrhiza uralensis.

[0050] Example 7

[0051] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 16 parts of processed Aconitum carmichaelii, 8 parts of processed Pinellia ternata, 6 parts of Aster tataricus, 8 parts of Curcuma zedoaria, 6 parts of Poria cocos, 5 parts of Paeonia suffruticosa, 5 parts of processed Evodia rutaecarpa, 10 parts of ginseng, 5 parts of Zanthoxylum bungeanum, and 5 parts of Glycyrrhiza uralensis.

[0052] Example 8

[0053] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 14 parts of processed Aconitum carmichaelii, 6 parts of processed Pinellia ternata, 6 parts of Aster tataricus, 6 parts of Curcuma zedoaria, 6 parts of Poria cocos, 6 parts of Paeonia suffruticosa, 6 parts of processed Evodia rutaecarpa, 8 parts of ginseng, 4 parts of Zanthoxylum bungeanum, and 4 parts of Glycyrrhiza uralensis.

[0054] Experimental Example 1

[0055] Effects of the traditional Chinese medicine composition of this invention on the proliferation, migration and apoptosis of A549 lung cancer cells

[0056] 1 Experimental Methods

[0057] 1.1 Preparation of drug-containing serum

[0058] Twenty SPF-grade SD rats (half male, half female, weighing (200±20) g) were randomly divided into two groups of 10 rats each: a control group and a traditional Chinese medicine (TCM) group. The TCM group received 1 mL / 100 g of the oral TCM solution prepared in Example 8 via gavage, plus an intraperitoneal injection of an equal volume of physiological saline. The control group received an equal volume of physiological saline via gavage, plus an intraperitoneal injection of an equal volume of physiological saline. Both groups received the TCM treatment via gavage for 7 consecutive days. Two hours after the last administration, the rats were anesthetized with 10% chloral hydrate via intraperitoneal injection (IP), and blood was collected from the abdominal aorta under sterile conditions. The blood was allowed to stand at 4°C for 3 hours, and then centrifuged to separate the serum. The serum from each group was combined and inactivated in a 56°C water bath for 30 minutes. The serum was then filtered through a 0.22 μm needle filter for sterilization, aseptically dispensed into 1 mL EP tubes, and stored at -20°C for later use.

[0059] 1.2 Cell Culture

[0060] A549 lung cancer cell suspension was prepared, and A549 lung cancer cells were cultured using standard cell culture methods. The cells were then expanded to passages 6-8 with good growth for experiments. The cells were divided into 3 groups, with 5 replicates per group and 1×10⁻⁶ wells. 6 Cells were seeded per well in 96-well plates and cultured in BPMI 1640 medium containing 10% fetal bovine serum for 24 hours. The medium was then aspirated, and 200 μL of serum-free BPMI 1640 medium was added to each well. Cells were cultured for another 24 hours to synchronize A549 lung cancer cells.

[0061] 1.2 Proliferation of A549 lung cancer cells

[0062] A549 lung cancer cell suspension was prepared, and A549 lung cancer cells were cultured using standard cell culture methods. The cells were then expanded to passages 6-8 with good growth for experiments. The cells were divided into 3 groups, with 5 replicates per group and 1×10⁻⁶ wells. 6 Cells were seeded per well in 96-well plates and cultured in BPMI 1640 medium containing 10% fetal bovine serum for 24 hours. The medium was then aspirated, and 200 μL of serum-free BPMI 1640 medium was added to each well. Cells were cultured for another 24 hours to synchronize A549 lung cancer cells. A549 lung cancer cells were then cultured in BPMI 1640 containing 10% control mouse serum, serum from the traditional Chinese medicine group, and cisplatin (5 μg / mL), 200 μL per well. After 24 hours of incubation, 10 μL of CCK8 was added and the cells were cultured for 1–4 hours. The absorbance at 450 nm was then measured, and the cell proliferation rate was calculated.

[0063] Cell proliferation rate = (Absorbance of experimental group / Absorbance of blank control group) × 100%.

[0064] 1.3 Migration of A549 lung cancer cells

[0065] Use a marker pen to draw three evenly spaced horizontal lines on the back of the 6-hole board; add approximately 1×10 [units of material] into the holes. 5 A549 cells were cultured until the cell density reached over 90%. Using a 200 μL pipette tip, aligned with a ruler, vertical lines were drawn along the horizontal lines on the back of the cell. Damaged and detached cells were washed away with PBS. Drug intervention was then performed using the drug-containing serum prepared in section 1.1, with the serum added at 10% of the culture medium volume. Cells were returned to the incubator and cultured for 24 hours. Samples were taken and migration rate was calculated as follows: Cell migration rate = (0h scratch width - 24h scratch width) / 0h scratch width × 100%.

[0066] 1.4 Apoptosis in A549 lung cancer cells

[0067] Cell apoptosis was detected by flow cytometry. Specifically, cells with a concentration of 1×10⁶ cells were used. 5 A549 cell suspension was seeded at 2 mL / well in 6-well plates. After incubation at 37°C and 5% CO2 for 24 h, the culture medium was discarded, and drug intervention was performed according to the group. The cells were cultured for another 48 h, collected by centrifugation (300×g, 5 min), and resuspended in 500 μL of 1×Annexin V-Binding Buffer. 5 μL each of Annexin V-APC and 7-AAD were added, gently mixed, and incubated at room temperature in the dark for 15–20 min. The cells were then immediately analyzed.

[0068] 2 Results

[0069] 2.1 Effects of the herbal combination of the present invention on the proliferation of A549 lung cancer cells

[0070] Compared with the control group, the proliferation rate of cancer cells in the traditional Chinese medicine group and the cisplatin group was significantly reduced (P<0.01), indicating that the traditional Chinese medicine composition of this invention has a significant inhibitory effect on the proliferation of A549 lung cancer cells, and its inhibitory effect on cancer cell proliferation is comparable to that of cisplatin. The results are as follows... Figure 1 As shown.

[0071] 2.2 Effects of the herbal composition of the present invention on the migration ability of A549 lung cancer cells

[0072] Compared with the control group, the cancer cell migration rate in the traditional Chinese medicine group and the cisplatin group was significantly reduced (P<0.01), indicating that the traditional Chinese medicine composition of this invention has a significant inhibitory effect on the migration of A549 lung cancer cells, and its inhibitory effect on cancer cell migration is comparable to that of cisplatin. The results are as follows... Figure 2 As shown.

[0073] 2.3 Effects of the herbal composition of the present invention on apoptosis of A549 lung cancer cells

[0074] Compared with the control group, the apoptosis rate of cancer cells in the traditional Chinese medicine group and the cisplatin group was significantly increased (P<0.01); compared with the cisplatin group, the apoptosis rate of cancer cells in the traditional Chinese medicine group was significantly increased (P<0.05). This indicates that the traditional Chinese medicine composition of the present invention has a significant promoting effect on apoptosis of A549 lung cancer cells, and the effect of promoting cancer cell apoptosis is significantly better than that of cisplatin. The results are as follows... Figure 3 As shown.

[0075] Based on the above experimental results, the traditional Chinese medicine composition of the present invention can effectively inhibit the proliferation and migration of cancer cells and promote apoptosis of cancer cells, providing a theoretical basis for the use of the traditional Chinese medicine composition of the present invention in the preparation of drugs for treating lung cancer.

[0076] Experimental Example 2

[0077] Effects of the herbal composition of this invention on Lewis lung cancer model mice

[0078] 1. Materials and Methods

[0079] 1.1 Experimental animals and Lewis lung cancer cell line

[0080] Thirty male SPF-grade C57BL / 6 mice, aged 6-8 weeks and weighing (20±3) g, were purchased from Spiford (Beijing) Biotechnology Co., Ltd. Lewis lung cancer cell lines were provided by the Cell Bank of the Chinese Academy of Sciences.

[0081] 1.2 Test Drugs

[0082] The oral liquid of traditional Chinese medicine of the present invention was prepared according to Example 8.

[0083] Cisplatin Injection, a product of Qilu Pharmaceutical Co., Ltd., product batch number EA1A9004B.

[0084] 1.3 Establishment and Grouping of Animal Models

[0085] Thirty mice were randomly divided into a model control group, a traditional Chinese medicine group, and a cisplatin group, with 10 mice in each group. A mouse model of lung cancer was established using subcutaneous inoculation: Lewis cells cultured in DMEM high-glucose medium (containing 10% fetal bovine serum) were digested with trypsin, counted, and then prepared into 1×10⁶ cells using physiological saline. 7Cell suspension at 0.2 mL / mL was administered subcutaneously to the right axilla of each C57BL / 6 mouse. Mice were returned to their cages after inoculation. One day after inoculation, the mice underwent the following interventions: Traditional Chinese Medicine Group: 1 mL / 100g oral liquid of the present invention was administered by gavage, followed by an intraperitoneal injection of an equal volume of physiological saline; Cisplatin Group: An equal volume of physiological saline was administered by gavage, followed by an intraperitoneal injection of 0.5 μg / (g·d) cisplatin; Model Group: An equal volume of physiological saline was administered by gavage, followed by an intraperitoneal injection of an equal volume of physiological saline. Gavage and intraperitoneal injection were administered once daily for 14 days.

[0086] 1.4 Detection Indicators

[0087] 1.4.1 General Case

[0088] During the drug administration period, the fur color, mental state, diet, and activity of mice in each group were observed at fixed times.

[0089] 1.4.2 Tumor growth status

[0090] Twenty-four hours after the last administration, mice were euthanized by cervical dislocation, and tumor tissue was removed, measured, and weighed.

[0091] 1.4.3 Tumor histopathological morphology

[0092] Mouse tumor tissue was taken, fixed with 40 g / L paraformaldehyde, routinely embedded in paraffin, and 4 μm thick sections were prepared. Hematoxylin-eosin staining was performed, and the morphological changes of the tumor tissue were observed under a microscope.

[0093] 1.4.4 Ki-67 expression in tumor tissue

[0094] Mouse tumor tissue was collected, embedded in paraffin, and 4 μm thick sections were prepared. Immunohistochemistry was used to detect the expression of the cell proliferation protein Ki-67. The positive staining of the tissue was observed under a microscope, and the optical density value was analyzed using Image-Pro Plus 6.0 software.

[0095] 1.5 Statistical Analysis

[0096] SPSS 18.0 statistical analysis software was used for data processing. Data are presented as mean values. Mean standard deviation (s) is expressed as mean ± standard deviation. One-way ANOVA was used for comparisons between groups. The Bonferroni test was used when variances were homogeneous, and the Dunnett's T3 test was used when variances were unequal. The significance level was set at α = 0.05.

[0097] 2 Results

[0098] 2.1 General condition of mice in each group

[0099] The model group exhibited a series of poor symptoms, including lethargy, slow reaction, sluggish movement, dull and yellow fur, and reduced food and water intake. The mice in the traditional Chinese medicine group were in good overall condition, showing agile reactions, glossy and smooth fur, and normal appetite; while the mice in the cisplatin group showed reduced appetite and water intake, weight loss, and dull fur.

[0100] 2.2 Tumor volume and weight in each group of mice

[0101] Compared with the model group, the tumor mass and tumor volume of mice in the traditional Chinese medicine group and cisplatin group were significantly reduced (P<0.05, P<0.01), as shown in the results. Figure 4 As shown, this indicates that the herbal composition of the present invention has a significant inhibitory effect on tumor growth.

[0102] 2.3 Pathological morphology of tumor tissues in mice of each group

[0103] HE staining of tumor tissue from model mice showed that tumor cells had large, deeply stained nuclei, were densely packed, and had abnormal morphology, with frequent mitotic figures and few necrotic areas. In contrast, the tumor tissues from the traditional Chinese medicine group and the cisplatin group showed relatively sparse cell arrangement, with a few scattered vacuolated cells, disordered arrangement in some areas, and numerous areas of necrotic cells. The results are as follows: Figure 5 As shown, this indicates that the herbal composition of the present invention can improve the pathological morphology of tumor tissue.

[0104] 2.4 Ki-67 expression in tumor tissues of mice in each group

[0105] The presence of numerous brown granules in the tumor tissue of the model group mice indicates a high proportion of Ki-67 positivity, a high expression level, and strong cancer cell proliferation ability. Figure 6 Compared with the model group, the number of brown granules in the tumor tissues of mice in the traditional Chinese medicine group and the cisplatin group was significantly reduced, and the proportion of Ki-67 positivity was significantly decreased (P<0.05, P<0.01). The results are as follows: Figure 4 As shown, this indicates that the herbal composition of the present invention can significantly inhibit the expression of Ki-67 and has a significant inhibitory effect on the proliferation activity of tumor cells.

[0106] 3. Conclusion

[0107] The traditional Chinese medicine composition of this invention has a significant therapeutic effect on tumor model mice, significantly improving their survival status. It also significantly inhibits the expression of the cell proliferation marker Ki-67, thus achieving a significant inhibitory effect on tumor growth and significantly improving the pathological morphology of tumor tissue. This traditional Chinese medicine composition provides a new direction for the development of drugs for the treatment of lung cancer.

[0108] Application examples

[0109] Mr. Wang, male, 69 years old. First visit on August 3, 2022. Symptoms included cough with yellow phlegm, occasional chest tightness and shortness of breath, dark complexion, stomach discomfort, poor mental state, sallow complexion, dark red tongue with thin yellow coating, and thready and rapid pulse.

[0110] Chest CT scan showed a mass in the lower lobe of the right lung, suggestive of malignancy. A transbronchial biopsy was performed, and postoperative pathology revealed squamous cell carcinoma, moderately differentiated. Immunohistochemistry showed PD-L1 (TPS: 90%) and Ki67 (approximately 50%+).

[0111] Diagnosis: Squamous cell carcinoma of the right lung (T2N2M0, stage IIIa).

[0112] On August 10, 2022, the patient underwent CT-guided radioactive particle implantation combined with tislelizumab for treatment. Starting September 6, 2022, in addition to Western medicine treatment, the patient took the traditional Chinese medicine pills prepared in Example 8 orally, 10 pills three times a day. Regular follow-ups were conducted. During the medication period, the patient reported significant improvement in cough and sputum production compared to before medication, reduced heaviness and fatigue, good appetite, and a significant improvement in quality of life, with no chest tightness, shortness of breath, or other discomfort. Six months after medication, the patient returned for a follow-up visit on March 6, 2023. The patient's mental state was fair, with a pale red tongue, thin yellow coating, and a thready, rapid pulse. Chest CT scans before and six months after treatment showed that the tumor lesion in the lower lobe of the right lung had significantly shrunk compared to before treatment, and the multiple calcified nodules and linear shadows in both lungs had also shrunk. Tumor marker levels showed a 48.44% decrease in neuron-specific enolase (NSE) and a 35.14% decrease in cytokeratin 19 fragment Cyfra21-1.

[0113] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A traditional Chinese medicine composition for treating lung cancer, characterized in that, The traditional Chinese medicine composition is made from the following traditional Chinese medicine raw materials in parts by weight: 5-20 parts of processed Aconitum carmichaelii, 3-15 parts of processed Pinellia ternata, 3-15 parts of Aster tataricus, 3-15 parts of Curcuma zedoaria, 3-15 parts of Poria cocos, 3-15 parts of Paeonia suffruticosa, 3-15 parts of processed Evodia rutaecarpa, 3-15 parts of Panax ginseng, 3-15 parts of Zanthoxylum bungeanum, and 3-15 parts of Glycyrrhiza uralensis.

2. The traditional Chinese medicine composition for treating lung cancer according to claim 1, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 10-20 parts of processed Aconitum carmichaelii, 3-10 parts of processed Pinellia ternata, 3-10 parts of Aster tataricus, 3-10 parts of Curcuma zedoaria, 3-10 parts of Poria cocos, 3-10 parts of Paeonia suffruticosa, 3-10 parts of processed Evodia rutaecarpa, 5-15 parts of Panax ginseng, 3-10 parts of Zanthoxylum bungeanum, and 3-10 parts of Glycyrrhiza uralensis.

3. The traditional Chinese medicine composition for treating lung cancer according to claim 1, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 14 parts of processed Aconitum carmichaelii, 6 parts of processed Pinellia ternata, 6 parts of Aster tataricus, 6 parts of Curcuma zedoaria, 6 parts of Poria cocos, 6 parts of Paeonia suffruticosa, 6 parts of processed Evodia rutaecarpa, 8 parts of ginseng, 4 parts of Zanthoxylum bungeanum, and 4 parts of Glycyrrhiza uralensis.

4. The use of the traditional Chinese medicine composition according to any one of claims 1 to 3 in the preparation of a medicament for treating lung cancer, characterized in that, The drug is prepared into an oral dosage form using the above-mentioned proportions of traditional Chinese medicine as raw materials and supplemented with medically acceptable excipients. The dosage form is any one of powder, granules, pills, decoction, powder, oral liquid, tablets, and capsules.

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