Traditional Chinese medicine composition for treating lung cancer as well as preparation method and application thereof

Abstract

The invention provides a traditional Chinese medicine composition for treating lung cancer. The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 35-45 parts of wild buckwheat rhizome, 25-35 parts of marsdenia tenacissima, 15-25 parts of radix ranunculi ternati, 25-35 parts of astragalus membranaceus, 15-25 parts of glossy privet fruit, 25-35 parts of caulis spatholobi and 5-15 parts of platycodon grandiflorum. The invention further provides a preparation method and application of the traditional Chinese medicine composition. The pharmaceutical composition disclosed by the invention is suitable for treating cancers, such as lung cancer, and is particularly suitable for treating phlegm-blood stasis and qi-yin deficiency type cancers; the traditional Chinese medicine composition only contains seven traditional Chinese medicines, the medicines are proper, the raw materials are rich and easy to obtain, the cost is low, and the traditional Chinese medicine composition is suitable for popularization and application.

Description

Technical Field

[0001] The present invention relates to a traditional Chinese medicine composition for treating lung cancer, its preparation method and uses, and belongs to the field of traditional Chinese medicine. Background Art

[0002] Lung cancer is one of the malignant tumors that seriously endanger human health. In recent years, its incidence has been continuously rising around the world and has ranked first among malignant tumors in many large cities. The causes of lung cancer are very complex and are closely related to smoking, air pollution, chronic respiratory diseases, family genetics, mental factors, etc.

[0003] Lung cancer is a common disease name in both traditional Chinese and Western medicine. Although the disease name "lung cancer" did not appear in ancient Chinese medical literature of past dynasties, according to its clinical manifestations and descriptions of similar diseases in ancient books, it can be classified into the categories of "lung accumulation", "xiben", "xiji", "pijie", "lung atrophy", "lung abscess", "lung carbuncle", "consumptive cough", etc. As early as in "The Classic of Difficult Issues - Fifty-Sixth Difficulty", it was pointed out that "the accumulation in the lung is called xiben, which is located under the right hypochondrium and is as large as a cup when covering. If it persists for a long time, it will cause aversion to cold and fever, asthma and cough, and develop into lung abscess". "Plain Questions - Theory of Strange Diseases" states: "A person has a disease with fullness under the hypochondrium and qi reversal, which has not healed for two or three years... The disease is called xiji... Medicine alone cannot cure it". What is recorded in "Spiritual Pivot - Pathogenic Factors and Diseases of the Zang-Fu Organs" is: "When the lung pulse is extremely tense, it is epilepsy; when it is slightly tense, it is lung cold and heat, listlessness, coughing up blood, and pain radiating to the waist, back and chest". "Plain Questions - Theory of Genuine Manifestation of the Viscera" states: "The large bones are withered and emaciated, the large muscles are sunken, there is fullness of qi in the chest, dyspnea and discomfort, and internal pain radiating to the shoulder and neck". The described cough, qi reversal and dyspnea, hemoptysis, cachexia, etc. are all similar to the symptoms of lung cancer. After the Han Dynasty, with the continuous enrichment of traditional Chinese medical theory and clinical practice, the understanding of lung cancer became more in-depth. "General Treatise on the Causes and Symptoms of Various Diseases" in the Sui Dynasty recorded: "This is caused by the accumulation of drinking water that does not disperse, and then combined with diet... or distension and pain, or asthma and shortness of breath, so it is called pijie". "Prescriptions for Universal Relief" in the Song Dynasty said: "The condition of xiben is under the right hypochondrium and is as large as a cup when covering, with rapid breathing and overflowing qi, which is lung accumulation". "Complete Collection of Holy Benevolent Medicine" also states: "Lung accumulation, xiben, qi fullness and distension, coughing, sniveling and expectorating pus and blood". "Compendium of Medicine" in the Ming Dynasty mentioned: "Consumptive cough, hoarseness, inability to make a sound or asthma and shortness of breath, this is the failure of the lung zang-organs and will surely lead to death". "Standards for Diagnosis and Treatment of Miscellaneous Diseases" said: "Lung atrophy, either coughing up foam or coughing up blood". "Dongyi Baojian - Carbuncle and Ulcer" in the Ming Dynasty stated: "When carbuncle and ulcer occur internally, the zang-organs should be examined. If there is a dull pain in Zhongfu, it is lung carbuncle", and for the first time, lung cancer was defined as "internal carbuncle".

[0004] Lung cancer has its origin in the lungs and involves the spleen and kidneys, belonging to the syndrome of deficiency in origin and excess in superficiality. Insufficient vital qi is the basis for the onset of lung cancer; invasion by the six exogenous pathogenic factors of wind, cold, summer heat, dampness, dryness, heat or epidemic pathogenic toxins is a common inducing factor for the onset of lung cancer; the tumor mass in the lungs is one of the main local manifestations of lung cancer. Just as stated in "The Spiritual Pivot - The Beginning of All Diseases": "Internal injury by worry and anger causes qi to reverse upward. When qi reverses upward, the six transport channels are blocked, the warm qi cannot circulate, the coagulated blood accumulates internally and does not disperse, the body fluids ooze and stagnate, adhere and do not leave, and all accumulations are formed." Another example is from "Medical Classics for Necessary Reading - Accumulation and Aggregation": "The formation of accumulations is due to insufficient vital qi, and then pathogenic qi occupies it." Internal injury by the seven emotions, unclean diet, abnormal work and rest, etc. lead to disharmony of the functions of the zang-fu organs in the human body, resulting in a series of pathological changes such as qi stagnation, blood stasis, toxin accumulation, damp stagnation, and phlegm coagulation, and finally accumulations form tumors. Therefore, although the occurrence of lung cancer is complex and diverse, its causes are nothing more than deficiency and excess, attributed to the interaction between internal factors (deficiency of vital qi) and external factors (the six exogenous pathogenic factors, internal injury by the seven emotions, diet, fatigue). The basic pathogenesis is deficiency of vital qi and excess of pathogenic factors. Deficiency of vital qi is the internal basis for the occurrence of tumors. Pathogenic toxins invade the lungs, phlegm and stasis accumulate internally, and cancerous tumors develop.

[0005] According to the clinical manifestations of lung cancer, it can be divided into three types: mainly excess syndrome, mainly deficiency syndrome, and deficiency-excess complex syndrome. The excess syndromes are mostly phlegm-dampness accumulating in the lungs type, qi stagnation and blood stasis type, phlegm and blood stasis mutually binding type, phlegm-heat accumulating in the lungs type; the deficiency syndromes are mostly yin deficiency with internal heat type, qi and yin deficiency type, qi and blood deficiency type, yin and yang deficiency type; the deficiency-excess complex syndromes are mostly spleen deficiency and qi stagnation type, qi deficiency and phlegm coagulation type, qi deficiency and blood stasis type, yin deficiency and phlegm-heat type, qi deficiency and phlegm-heat type, etc. In clinical treatment, according to the bias of deficiency of vital qi and excess of pathogenic factors, dispelling pathogenic factors while supplementing vital qi, supplementing vital qi while dispelling pathogenic factors, or equally emphasizing supplementing vital qi and dispelling pathogenic factors are adopted. Common treatment principles include: the method of strengthening the healthy qi and cultivating the primordial qi, the method of promoting blood circulation to remove blood stasis, the method of clearing heat and detoxifying, the method of promoting qi movement to relieve stagnation, the method of softening hardness and dissipating nodules, etc., all of which need to be flexibly selected according to different stages and manifestations of the disease. Just as stated in the chapter on accumulation and aggregation in "Medical Classics for Necessary Reading", the three principles of treating at the initial, middle, and final stages are: "In the initial stage, when the pathogenic qi has just arisen, the vital qi is still strong and the pathogenic qi is still shallow, then it can withstand attacking; in the middle stage, when the disease has lasted for some time, the pathogenic qi is deeper and the vital qi is weaker, it can withstand both attacking and supplementing; in the final stage, when the disease has lasted for a long time, the pathogenic qi invades and the vital qi is depleted, then it can withstand supplementing." It is pointed out that in the initial stage, attacking and dispersing should be carried out, in the middle stage, both eliminating and supplementing should be combined, and in the final stage, when the deficiency of vital qi is obvious and the pathogenic accumulation is deep, supplementation should be combined with elimination, and nourishing the healthy qi to eliminate accumulation.

[0006] As a malignant tumor that seriously threatens human health, lung cancer has become a social public health problem that needs to be solved urgently due to its high incidence, high mortality rate, serious impact on patients' quality of life and high treatment costs. In the existing treatment of lung cancer, commonly used drugs are chemotherapy drugs, targeted drugs and immunosuppressants. After use, the above drugs have certain toxic and side effects on patients, and the drugs are prone to drug resistance after a certain period of use, and there are obvious treatment limitations. In contrast, traditional Chinese medicine treatment can effectively improve the quality of life of lung cancer patients, improve clinical symptoms, and prolong the life cycle. It is widely used in the clinical treatment of advanced lung cancer. Its multi-target, multi-pathway, and overall regulatory characteristics in the treatment of lung cancer, as well as its wide range of applicable populations, low toxic and side effects, high safety, good tolerance and low economic cost are increasingly attracting attention and occupy an important position in the treatment of lung cancer.

[0007] Application number: CN97116956.X, invention name: a method for preparing a golden buckwheat preparation, the application discloses that the ethanol extract of golden buckwheat is used to treat lung cancer, gastric cancer, cervical cancer, and to prepare anti-inflammatory drugs, drugs for reducing the toxic and side effects of tumor radiotherapy and chemotherapy, and drugs that have a synergistic effect on tumor radiotherapy and chemotherapy. Application number CN200410040094.1, invention name: compound golden buckwheat preparation, is composed of extracts of Chinese herbal medicines golden buckwheat, bergenia and Gynostemma pentaphyllum. The extraction method of golden buckwheat powder is: take golden buckwheat rhizomes, wash and slice, dry them, soak them in acetone-water solvent at room temperature for 2-7 days, reduce the pressure of the extract to recover acetone, concentrate and filter to remove precipitates, extract with ether: ethyl acetate: n-butanol, use D100 macroporous adsorption resin as filler, use ethanol of different concentrations as eluent, separate ethyl acetate and n-butanol extracts, collect the eluent, let it stand for 24 hours, and dry it at 105℃. The preparation has the effects of invigorating qi and nourishing yin, clearing away heat and detoxifying, promoting blood circulation and removing blood stasis, and removing phlegm and relieving cough; it has cancer chemoprevention and anticancer activity, and can be used to treat lung cancer. CN201711223794.8, invention name: A kind of anti-cancer pills of a pharmaceutical composition containing scutellaria baicalensis and a preparation method, relates to a kind of pills of a pharmaceutical composition containing scutellaria baicalensis and a preparation method. The pharmaceutical composition is made of scutellaria baicalensis, cinnamon, ephedra and mistletoe. The pharmaceutical composition and preparation containing scutellaria baicalensis provided by the present invention, especially the pills, have excellent effects in resisting various tumors, such as liver cancer, gastric cancer and esophageal cancer, lung cancer, cervical cancer and leukemia malignant tumors. Because the clinical manifestations, etiology and pathogenesis of lung cancer are very complicated, the existing reported prescriptions still cannot meet clinical needs. Summary of the invention

[0008] The technical solution of the present invention is to provide a new Chinese medicine composition for treating lung cancer. Another technical solution of the present invention provides a preparation method and use of the Chinese medicine composition.

[0009] The present invention provides a traditional Chinese medicine composition for treating lung cancer, which is prepared from raw medicinal materials in the following weight ratios:

[0010] 35 - 45 parts of Fagopyrum dibotrys, 25 - 35 parts of Marsdenia tenacissima, 15 - 25 parts of Ranunculus ternatus Thunb., 25 - 35 parts of Astragalus membranaceus, 15 - 25 parts of Glossy Privet Fruit, 25 - 35 parts of Spatholobus suberectus, 5 - 15 parts of Platycodon grandiflorum.

[0011] More preferably, it is prepared from raw medicinal materials in the following weight ratios:

[0012] 40 parts of Fagopyrum dibotrys, 30 parts of Marsdenia tenacissima, 20 parts of Ranunculus ternatus Thunb., 30 parts of Astragalus membranaceus, 20 parts of Glossy Privet Fruit, 30 parts of Spatholobus suberectus, 10 parts of Platycodon grandiflorum.

[0013] Among them, the Glossy Privet Fruit is the processed Glossy Privet Fruit with wine.

[0014] The traditional Chinese medicine composition of the present invention takes the raw medicinal powder of the above - mentioned raw medicinal materials, water or organic solvent extract as the active ingredient, and adds pharmaceutically acceptable excipients or auxiliary components to prepare commonly used oral preparations in pharmacy.

[0015] Among them, the oral preparations are extractives, pills, dripping pills, granules, tablets, powders, capsules, tea preparations, oral liquids, syrups.

[0016] The present invention provides a method for preparing the above - mentioned traditional Chinese medicine composition for treating lung cancer, which comprises the following steps:

[0017] a. Weigh the raw medicinal materials in each weight ratio;

[0018] b. Directly powder the raw medicinal materials, extract them with water or organic solvents, and add pharmaceutically acceptable excipients or auxiliary components to prepare commonly used oral preparations in pharmacy.

[0019] The present invention also provides the use of the above - mentioned traditional Chinese medicine composition in the preparation of drugs for treating lung cancer.

[0020] More preferably, the lung cancer is non - small cell lung cancer.

[0021] More preferably, the lung cancer is the type of lung cancer with the syndrome of intermingled phlegm and stasis and deficiency of both qi and yin.

[0022] More preferably, the drug is a drug with the effects of shrinking tumors, inhibiting tumor growth, preventing tumor metastasis, and preventing tumor recurrence.

[0023] The present invention also provides the use of the above - mentioned traditional Chinese medicine composition in the preparation of drugs with the effects of strengthening healthy qi and tonifying deficiency, dissipating stasis and resolving masses, and strengthening the kidney and invigorating the spleen.

[0024] The raw material sources of the traditional Chinese medicine composition of the present invention are as follows:

[0025] Fagopyrum dibotrys (D. Don) Hara, a plant of the Polygonaceae family, is recorded in the first volume of the Chinese Pharmacopoeia 2020 Edition on pages 228 - 229. Its dried rhizome has a slightly pungent, astringent and cool taste and belongs to the lung meridian.

[0026] Marsdenia tenacissima (Roxb.) Wight et Arn., a plant of the Asclepiadaceae family, is recorded in the first volume of the Chinese Pharmacopoeia 2020 Edition on page 309. Its dried vine stem is bitter and slightly cold and belongs to the lung meridian.

[0027] Ranunculus ternatus Thunb., a plant of the Ranunculaceae family, is recorded in the first volume of the Chinese Pharmacopoeia 2020 Edition on page 333. Its dried tuberous root is sweet, pungent and warm and belongs to the liver and lung meridians.

[0028] Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao or Astragalus membranaceus (Fisch.) Bge., plants of the Leguminosae family, are recorded in the first volume of the Chinese Pharmacopoeia 2020 Edition on pages 315 - 316. Their dried roots are sweet and slightly warm and belong to the lung and spleen meridians.

[0029] Ligustrum lucidum Ait., a plant of the Oleaceae family, is recorded in the first volume of the Chinese Pharmacopoeia 2020 Edition on pages 47 - 48. Its dried ripe fruits are sweet, bitter and cool and belong to the liver and kidney meridians.

[0030] Spatholobus suberectus Dunn, a plant of the Leguminosae family, is recorded in the first volume of the Chinese Pharmacopoeia 2020 Edition on pages 202 - 203. Its dried vine stem is bitter, sweet and warm and belongs to the liver and kidney meridians.

[0031] Platycodon grandiflorum (Jacq.) A. DC., a plant of the Campanulaceae family, is recorded in the first volume of the Chinese Pharmacopoeia 2020 Edition on page 289. Its dried root is bitter, pungent and neutral and belongs to the lung meridian.

[0032] Based on over thirty years of clinical experience, the inventor analyzed the pathogenesis and syndromes of lung cancer and summarized that the intermingling of phlegm and stasis, and deficiency of both qi and yin are one of the most common syndromes in the middle and late stages of lung cancer and after surgery. The traditional Chinese medicine composition of the present invention mainly dispels pathogenic factors and supplements healthy qi as an auxiliary. In the formula, Fagopyrum dibotrys is slightly pungent, astringent, and cool, and belongs to the lung meridian. It clears heat and detoxifies, expels pus and removes stasis, acting on toxin accumulation and blood stasis, and is the monarch drug. Marsdenia tenacissima is bitter and slightly cold, and belongs to the lung meridian. It softens hardness and dissipates nodules, detoxifies, acting on toxin accumulation, and is the ministerial drug. Ranunculus ternatus is sweet, pungent, and warm, and belongs to the liver and lung meridians. It resolves phlegm and dissipates nodules, detoxifies and reduces swelling, acting on phlegm coagulation masses, and is the ministerial drug. Astragalus membranaceus is sweet and slightly warm, and belongs to the spleen and lung meridians. It invigorates qi and raises yang, strengthens the exterior defense and consolidates the superficial resistance, expels toxins and discharges pus, and is used to raise yang and tonify the deficiency of qi in the spleen and lung, and improve immunity; Ligustrum lucidum is sweet, bitter, and cool, and belongs to the liver and kidney meridians. It benefits the liver and nourishes the kidney, clears deficiency heat, and is used to supplement the deficiency of yin in the liver and kidney; the main pathogenesis of tumors is the coexistence of deficiency in origin and excess in superficiality, especially deficiency of the spleen and kidney. The combination of Astragalus membranaceus and Ligustrum lucidum not only tonifies the deficiency of the spleen and lung, but also tonifies the deficiency of the liver and kidney, and has a special effect in tonifying the spleen for those with deficiency of the lung and spleen, and is a drug pair, assisting the monarch drug to achieve the effect of dispelling pathogenic factors and strengthening healthy qi, and at the same time tonifying the deficiency of the body caused by pathogenic toxins, and both are adjuvant drugs. Moreover, in addition to dispelling pathogenic factors and strengthening healthy qi, the composition of the present invention uses Caulis Spatholobi, a blood-activating drug, to promote blood circulation and nourish blood, acting on blood stasis and blood deficiency, and is an adjuvant drug. Platycodon grandiflorum is bitter, pungent, and neutral, and belongs to the lung meridian. It disperses the lung qi and eliminates phlegm, and is the guiding drug for the lungs and also the ferry of all drugs, and is the envoy drug. It has been proved by clinical application and experiments that it has the effects of inhibiting tumor growth, metastasis and preventing recurrence.

[0033] The beneficial effects of the present invention are as follows:

[0034] (1) The formula principle of the present invention is: treating both the principal and secondary aspects of a disease, with treating the secondary aspect as the main and supplementing the treatment of the principal aspect as an auxiliary; adopting the method of dispelling pathogenic factors and strengthening healthy qi, and tonifying the kidney and strengthening the spleen to enhance the immune function of patients. It can inhibit tumor growth and spread, prevent tumor recurrence, improve the quality of life of patients, and prolong the survival time of patients. The pharmaceutical composition of the present invention is applicable to the treatment of cancer, such as treating lung cancer; especially applicable to cancers with the syndrome of intermingling of phlegm and stasis, and deficiency of both qi and yin.

[0035] (2) The traditional Chinese medicine composition of the present invention only contains 7 traditional Chinese medicines, with refined ingredients, rich and easily available raw materials, and low cost, suitable for popularization and use. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 It is the tumor volume growth curve of animals in each experimental group during the experimental period (D0 - D21);

[0037] Figure 2 It is the relative tumor volume growth curve of animals in each experimental group during the experimental period (D0 - D21);

[0038] Figure 3 It is the influence of traditional Chinese medicine X on the proliferation rate of LLC transplanted tumors. DETAILED DESCRIPTION OF THE INVENTION

[0039] Example 1 Preparation of the Traditional Chinese Medicine Composition of the Present Invention

[0040] Raw materials: 40 g of Fagopyrum dibotrys, 30 g of Marsdenia tenacissima, 20 g of Ranunculus ternatus, 30 g of Astragalus membranaceus, 20 g of Glossy Privet Fruit processed with wine, 30 g of Spatholobus suberectus, 10 g of Platycodon grandiflorum.

[0041] Preparation method: Crush the seven medicinal materials of Fagopyrum dibotrys, Marsdenia tenacissima, Ranunculus ternatus, Astragalus membranaceus, Glossy Privet Fruit processed with wine, Spatholobus suberectus, and Platycodon grandiflorum, and then weigh the raw materials according to the above ratio; decoct twice. For the first time, add 1800 mL of water, bring to a boil over high heat for 10 minutes, and then continue to decoct over low heat for 30 minutes. For the second time, add 1400 mL of water, bring to a boil over high heat for 10 minutes, and then continue to decoct over low heat for 30 minutes. Combine the decoction liquids, concentrate under reduced pressure to an appropriate density, and dry under reduced pressure until the water content is about 6% or less to obtain the extract of the traditional Chinese medicine composition of the present invention (Traditional Chinese Medicine X).

[0042] Example 2 Preparation of the Traditional Chinese Medicine Composition of the Present Invention

[0043] Raw materials: 20 g of Fagopyrum dibotrys, 10 g of Marsdenia tenacissima, 10 g of Ranunculus ternatus, 10 g of Astragalus membranaceus, 10 g of Glossy Privet Fruit processed with wine, 10 g of Spatholobus suberectus, 3 g of Platycodon grandiflorum.

[0044] Preparation method: Crush the seven medicinal materials of Fagopyrum dibotrys, Marsdenia tenacissima, Ranunculus ternatus, Astragalus membranaceus, Glossy Privet Fruit processed with wine, Spatholobus suberectus, and Platycodon grandiflorum, and then weigh the raw materials according to the above ratio; decoct twice. For the first time, add 600 mL of water, bring to a boil over high heat for 10 minutes, and then continue to decoct over low heat for 30 minutes. For the second time, add 370 mL of water, bring to a boil over high heat for 10 minutes, and then continue to decoct over low heat for 30 minutes. Combine the decoction liquids, concentrate under reduced pressure to an appropriate density, and dry under reduced pressure until the water content is about 6% or less to obtain the extract of the traditional Chinese medicine composition of the present invention.

[0045] Example 3 Preparation of the Traditional Chinese Medicine Composition of the Present Invention

[0046] Raw materials: 60 g of Fagopyrum dibotrys, 50 g of Marsdenia tenacissima, 30 g of Ranunculus ternatus, 50 g of Astragalus membranaceus, 30 g of Glossy Privet Fruit processed with wine, 50 g of Spatholobus suberectus, 15 g of Platycodon grandiflorum.

[0047] Preparation method: Crush the seven medicinal materials of Fagopyrum dibotrys, Marsdenia tenacissima, Ranunculus ternatus, Astragalus membranaceus, Glossy Privet Fruit processed with wine, Spatholobus suberectus, and Platycodon grandiflorum, and then weigh the raw materials according to the above ratio; decoct three times. For the first time, add 4200 mL of water, bring to a boil over high heat for 10 minutes, and then continue to decoct over low heat for 30 minutes. For the second time, add 2300 mL of water, bring to a boil over high heat for 10 minutes, and then continue to decoct over low heat for 30 minutes. For the third time, add 1400 mL of water, bring to a boil over high heat for 10 minutes, and then continue to decoct over low heat for 30 minutes. Combine the decoction liquids, concentrate under reduced pressure to an appropriate density, and dry under reduced pressure until the water content is about 6% or less to obtain the extract of the traditional Chinese medicine composition of the present invention.

[0048] Example 4 Preparation of the Traditional Chinese Medicine Composition of the Present Invention

[0049] Raw materials: 15 g of Fagopyrum dibotrys, 10 g of Marsdenia tenacissima, 6 g of Ranunculus ternatus Thunb., 10 g of Astragalus membranaceus, 6 g of Glossy Privet Fruit processed with wine, 10 g of Caulis Spatholobi, 3 g of Platycodon grandiflorum.

[0050] Preparation method: Crush the seven medicinal materials of Fagopyrum dibotrys, Marsdenia tenacissima, Ranunculus ternatus Thunb., Astragalus membranaceus, Glossy Privet Fruit processed with wine, Caulis Spatholobi, and Platycodon grandiflorum, and then weigh the raw medicinal materials according to the above ratio; decoct once, add 600 mL of water, first bring to a boil over high heat for 10 minutes, then continue to decoct over low heat for 30 minutes, concentrate the decoction under reduced pressure to an appropriate density, and dry under reduced pressure until the water content is about 6% or less to obtain the extract of the traditional Chinese medicine composition of the present invention.

[0051] The beneficial effects of the present invention are demonstrated by the following specific pharmacodynamic experiments.

[0052] Experimental example 1 Experiment on the inhibitory effect of the traditional Chinese medicine composition of the present invention on the proliferation and growth of non-small cell lung cancer xenografts

[0053] This experiment verifies the inhibitory effect of the traditional Chinese medicine composition of the present invention on the proliferation and growth of non-small cell lung cancer xenografts and its tumor inhibitory and sensitizing effects under combined treatment with the positive control drug PD-1 antibody and cisplatin through in vivo experiments on animals.

[0054] 1 Experimental materials

[0055] 1.1 Experimental animals and cell lines

[0056] 90 male C57BL / 6J mice, SPF grade, purchased from Chengdu Yakang Biotechnology Co., Ltd., and the experimental animal certificate number is NO.511214900012706. The body weight of the animals was 15.96 - 19.89 g and the age was 6 - 7 weeks old when received; the body weight was 19.11 - 26.50 g and the age was 8 - 9 weeks old when grouped.

[0057] LLC cells (mouse primary Lewis lung cancer cells): purchased from the National Model and Characteristic Experimental Cell Resource Bank.

[0058] 1.2 Experimental drugs

[0059] Traditional Chinese medicine X prepared in Example 1; InvivoMab Anti mouse PD-1 antibody, manufactured by BioXcell; cisplatin injection, manufactured by Qilu Pharmaceutical Co., Ltd.

[0060] 1.3 Main experimental reagents and instruments

[0061] RPMI medium (batch number 8122055), Gibco TM Company; Trypsin cell digestive solution (batch number: 2323413), Gibco TM Company; Fetal bovine serum (batch number 2418958P), GibcoTM Company; PBS (batch number M210935), BasalMedia Company; Matrigel Matrix (batch number 2139002), Corning Company; 0.9% sodium chloride injection (batch number G221111702), Sichuan Kelun Pharmaceutical Co., Ltd.; CMC-Na, Sichuan GreenTech Biotechnology Co., Ltd.

[0062] One-percent balance (model ME2002), Mettler Toledo Technology (China) Co., Ltd.; One-thousandth balance (ME503T), Mettler Toledo Technology (China) Co., Ltd.; Biological safety cabinet (model 1300), Thermo Company.

[0063] 2 Experimental methods

[0064] 2.1 Preparation of test drugs

[0065] 2.1.1 Preparation of the test article of traditional Chinese medicine X

[0066] Low-dose treatment group of traditional Chinese medicine X: Weigh 12.5 g of the sample, grind and suspend it in 100 mL of 0.5% CMC-Na solution to prepare a test article with a concentration of 125 mg / mL;

[0067] High-dose treatment group of traditional Chinese medicine X: Weigh 37.5 g of the sample, grind and suspend it in 100 mL of 0.5% CMC-Na solution to prepare a test article with a concentration of 375 mg / mL;

[0068] 2.1.2 Positive control

[0069] PD-1 treatment group: Use the solvent (PBS) to prepare the PD-1 antibody to a concentration of 0.25 mg / mL;

[0070] Cisplatin treatment group: Dilute the positive control drug solution with the solvent (0.9% sodium chloride injection) to a concentration of 0.2 mg / mL;

[0071] 2.1.3 Negative control: 0.5% CMC-Na

[0072] 2.2 Modeling method

[0073] 2.2.1 Tumor cell culture and preparation

[0074] For the passage and culture of conventional tumor cell lines, the cells were passaged in vitro at least three times. When the murine lung cancer cells reached a confluence of more than 80%, the culture medium was discarded, and the cells were rinsed twice with PBS. Then, 0.25% trypsin was used for cell digestion, and the cells were allowed to stand for 1 - 2 minutes. Under the microscope, when the cell contours became round and cell clumps detached, complete medium containing 10% serum was added to terminate the digestion. The cells were collected by centrifugation at 1000 rpm for 3 minutes. The cells were washed twice with pre-cooled PBS, collected by centrifugation at 1000 rpm for 3 minutes, resuspended in PBS, counted using a cell counter, and the cell survival ratio was calculated. Only when the proportion of live cells exceeded 90% could the cells be used for the experiment. The cell concentration was adjusted to 1×10⁷ cells / mL. After cell collection, the cell suspension was placed on ice for pre-cooling. An appropriate amount of the cell suspension was mixed with an equal volume of Matrigel (Corning) on ice for cell seeding, and the cell seeding density was 1×10⁶ cells / animal.

[0075] 2.2.2 Subcutaneous tumor xenograft construction

[0076] Before cell seeding, the experimental animals were fixed, and the skin on the right hind leg was wiped with 75% alcohol. The cell suspension (200 μL) was injected subcutaneously. The needle was withdrawn slowly while rotating, and the injection site was pressed for half a minute to prevent cell leakage. After cell seeding, the inoculation site of the animals was continuously observed. When a lump formed, the long and short diameters of the tumor mass were measured using an electronic vernier caliper, and the tumor mass volume was calculated. When the average volume reached 100 mm 3 ³, the animals were randomly grouped according to the tumor volume.

[0077] 2.3 Group design

[0078] This experiment was divided into 9 groups. After tumor model establishment, the animals were randomly grouped according to the tumor volume, including a vehicle control group, an anti-PD-1 treatment group, a low-dose traditional Chinese medicine X group, a high-dose traditional Chinese medicine X group, a low-dose traditional Chinese medicine X + anti-PD-1 treatment group, a high-dose traditional Chinese medicine X + anti-PD-1 treatment group, a cisplatin treatment group, a low-dose traditional Chinese medicine X + cisplatin treatment group, and a high-dose traditional Chinese medicine X + cisplatin treatment group; 10 animals / group, a total of 90 animals, all male.

[0079] 2.4 Administration methods

[0080]

[0081]

[0082] Note: p.o. indicates oral gavage administration; i.v. indicates tail vein injection administration; i.p. indicates intraperitoneal injection administration.

[0083] 2.5 Observation and examination

[0084] 2.5.1 Tumor volume and tumor proliferation rate

[0085] Test animals: Experimental animals in each group;

[0086] Measurement method: 3 - 4 days after inoculation of tumor cells (when tumor nodules appear), use an electronic vernier caliper to measure the long diameter (a), short diameter (b) and height (c) of the tumor tissue of the experimental animals, calculate the tumor volume. After the average tumor volume reaches 250 mm3, randomly group the animals according to the tumor volume on that day and start the treatment with the test article. During the treatment period of the test article, use an electronic vernier caliper to measure the long diameter (a) and short diameter (b) of the tumor tissue, twice a week, calculate the tumor volume, plot the volume change curve, and calculate the relative tumor proliferation rate;

[0087] Calculation formula: Tumor volume (V) = π * a * b 2 / 6

[0088] Relative tumor proliferation rate T / C (%) = T RTV / C RTV ×100%;

[0089] RTV = V t / V0 (V0 is the tumor volume measured at the time of grouping and dosing, V t is the tumor volume at each measurement);

[0090] Measurement time: Twice a week, and measure once before gross dissection at the end of the experiment.

[0091] 2.5.2 Tumor weight and tumor growth inhibition rate

[0092] Test animals: Experimental animals in each group;

[0093] Measurement method: After the end of the drug treatment period, the experimental animals in each group are anesthetized by intraperitoneal injection of 3% pentobarbital sodium at a dose of 60 mg / kg, and then euthanized by exsanguination from the abdominal aorta. After that, take the tumor tissue and weigh the tumor weight for calculating the tumor growth inhibition rate (IR TW , %), and at the same time, take pictures of the tumor appearance with a digital camera;

[0094] Calculation formula:

[0095] Tumor growth inhibition rate = (average tumor weight of the control group - average tumor weight of the treatment group) / average tumor weight of the control group × 100%

[0096] Evaluation criteria: Refer to the relevant standards of the "Guidelines for the Research of Cytotoxic Antitumor Drugs".

[0097] 2.5.3 Gross dissection and anatomical observation

[0098] 2.5.3.1 Gross dissection

[0099] Dissection of animals: Animals in each experimental group

[0100] Dissection method: Animals were anesthetized with sodium pentobarbital (intraperitoneal injection, 60 mg / kg, and the dose could be adjusted appropriately according to the animal's health status). After collecting blood samples through the abdominal vena cava, gross dissection and anatomical observation of tumor tissues were performed;

[0101] Dissection time: End point of the experiment.

[0102] 3 Statistical analysis

[0103] The experimental data of each group of animals were expressed as mean ± standard deviation (Mean ± SD). For the comparison among multiple groups with normal distribution and homogeneous variance, one-way analysis of variance (ANOVA) was first used, and further multiple comparisons between groups were performed using Tukey's HSD test; if the data did not meet the normal distribution or the variance was not homogeneous, the Kruskal-Wallis H test (K-W method) was used for analysis, and multiple comparisons between groups were performed using Dunn's method. P < 0.05 was considered statistically significant. All statistical analyses were completed using Stata 15 software.

[0104] 4 Experimental results

[0105] 4.1 Effect of traditional Chinese medicine X on the growth of murine non-small cell lung cancer cell LLC xenografts

[0106] The measurement results of tumor volume during the experimental period (D0 - D21) ( Figures 1-2 ) showed that the volume of LLC xenografts in the tumor-bearing animals of the vehicle control group continued to increase. The average tumor volume increased from 108.79 ± 25.26 mm 3 on the day of grouping (D0) to 2194.09 ± 447.98 mm 3 at the end of the experiment (D21). The relative tumor volume (RTV) increased from 1.00 to 21.32 ± 7.95, indicating that the murine non-small cell lung cancer cell LLC subcutaneous xenograft model in this experimental system was successfully established and was suitable for the experimental research requirements of tumor proliferation inhibition.

[0107] The positive control drugs PD-1 antibody and cisplatin had obvious inhibitory effects on the proliferation and growth of LLC xenografts under the set treatment regimen (2.5 mg / kg and 2.0 mg / kg, intravenous injection through the tail vein and intraperitoneal injection, twice a week for 3 consecutive weeks). Their antitumor effects began to appear after the first week of treatment. The growth rate of tumor volume during the experimental period was significantly lower than that of the vehicle control group. The average tumor volume increased from 107.38 ± 17.08 mm 3 and 107.08 ± 20.28 mm 3 on the day of treatment grouping to 988.73 ± 398.85 mm 3and 1090.59 ± 331.00 mm 3 , the relative tumor volume increased from 1.00 to 9.40 ± 3.91 and 10.38 ± 3.33 respectively at the end of the experiment. The degree of tumor growth inhibition showed significant statistical differences compared with the tumor growth process of the vehicle control group animals starting from D7 (P < 0.05), and continued until the end of the experiment.

[0108] Traditional Chinese medicine X had a significant inhibitory effect on the proliferation and growth of LLC transplanted tumors under the set treatment regimen (2500 mg / kg and 7500 mg / kg, oral gavage once a day for 3 consecutive weeks). Compared with the vehicle control group in the same period, the tumor volume growth rates of the low and high-dose treatment groups of traditional Chinese medicine X in animals showed a slowdown. Its antitumor effect began to appear after the first week of treatment. The average tumor volume increased from 106.11 ± 20.01 mm 3 and 106.52 ± 18.24 mm 3 at the start of the treatment (D0) to 1124.30 ± 273.93 mm 3 and 1006.62 ± 333.40 mm 3 at the end of the experiment respectively. The relative tumor volume increased from 1.00 to 10.53 ± 2.06 and 9.23 ± 1.61 respectively at the end of the experiment. The degree of tumor growth inhibition showed significant statistical differences compared with the tumor growth process of the vehicle control group animals starting from D7 (P < 0.05), and continued until the end of the experiment. The antitumor degree was consistent with that of the positive control drugs PD-1 antibody and cisplatin, and no statistical differences were found (P > 0.05), but its antitumor effect showed no obvious dose correlation.

[0109] At the same time, no obvious sensitization effect was observed when traditional Chinese medicine X at low or high doses was combined with the positive control drugs PD-1 antibody and cisplatin. The tumor volume growth process of the animals in the combined treatment group was the same as that of the single drug treatment group in the same period. Among them, the tumor volumes of the animals in the low-dose and high-dose traditional Chinese medicine X combined with PD-1 antibody treatment groups were 108.11 ± 21.72 mm 3 and 108.04 ± 22.89 mm 3 at the start of the treatment and increased to 1066.75 ± 369.75 mm 3 and 1040.71 ± 246.01 mm 3 at the end of the experiment respectively. The relative tumor volume increased from 1.00 to 10.46 ± 5.14 and 9.96 ± 1.79 respectively at the end of the experiment. The tumor volumes of the animals in the low-dose and high-dose traditional Chinese medicine X combined with cisplatin treatment groups were 107.63 ± 21.01 mm3 and 107.53 ± 20.96 mm 3 at the start of the treatment and increased to 979.20 ± 377.40 mm 3and 1008.01 ± 177.54 mm 3 , the relative tumor volume increased from 1.00 to 9.05 ± 3.38 and 9.36 ± 2.62 at the end of the experiment. The degree of tumor growth inhibition showed significant statistical differences compared with the tumor growth process of the vehicle control group animals during the same period starting from D7 (P < 0.05), but no statistical differences were observed compared with the low or high dose of traditional Chinese medicine X, PD-1 antibody, and cisplatin treatment groups during the same period (P > 0.05).

[0110] Table 1: Effects of traditional Chinese medicine X on the growth of LLC xenograft tumors

[0111]

[0112] Note: All individual data are expressed as mean ± standard deviation (SD). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 indicates the statistical analysis results of the traditional Chinese medicine X, positive drug treatment group, or combined treatment group compared with the vehicle control group.

[0113] 4.2 Effects of traditional Chinese medicine X on the proliferation rate of murine non-small cell lung cancer cell LLC xenograft tumors

[0114] In this part of the experiment, the tumor proliferation processes of animals in each treatment group at different time points were analyzed based on the evaluation criteria of the "Technical Guidelines for Non-clinical Research of Cytotoxic Antitumor Drugs". The experimental results ( Figure 3 ) showed that during the experimental period, the tumors of the vehicle control group animals proliferated significantly. The positive control drugs PD-1 antibody and cisplatin had obvious inhibitory effects on the proliferation rate of LLC xenograft tumors under the set treatment regimens. Compared with the vehicle control group during the same period, the T / C% values showed a time-dependent decrease. At the end of the experiment, the levels were 49.40% and 48.69% respectively, and were greater than 40% during the experimental period.

[0115] Traditional Chinese medicine X also had a significant inhibitory effect on the proliferation rate of LLC xenograft tumors under the set treatment regimen. The T / C% values of the low and high dose treatment groups of animals showed a time-dependent decrease compared with the vehicle control group during the same period. At the end of the experiment, the levels were 43.28% and 49.06% respectively, and were greater than 40% during the experimental period. The degree of inhibition of LLC xenograft tumor proliferation was the same as that of the positive control drugs PD-1 antibody and cisplatin, and no differences were observed. At the same time, there was no obvious synergistic effect under the combined treatment conditions with PD-1 antibody and cisplatin. At the end of the experiment, the T / C% values of animals in each combined treatment group were the same as those under the single drug treatment conditions, and no differences were observed (P > 0.05).

[0116] 4.3 Effect of Traditional Chinese Medicine X on the Weight of Murine Non-Small Cell Lung Cancer Cell LLC Xenografts

[0117] At the end of the experiment, the tumor-bearing animals were euthanized, and the experimental results of dissecting and collecting tumor tissues for weighing showed that the positive control drugs, PD-1 antibody and cisplatin, had obvious inhibitory effects on the proliferation and growth of LLC xenografts under the set treatment regimen. Compared with the vehicle control group at the same time (3.360 ± 1.126 g), the tumor weights of the animals in the PD-1 antibody and cisplatin treatment groups showed significant decreases, which were 1.047 ± 0.318 g and 1.195 ± 0.337 g respectively, and the tumor growth inhibition rates (TGI) were 68.82% and 64.44% respectively. The differences were all statistically significant (P < 0.05).

[0118] Traditional Chinese Medicine X also had significant inhibitory effects on the proliferation and growth of LLC xenografts under the set treatment regimen. Compared with the vehicle control group at the same time, the tumors of the animals in the low and high dose treatment groups of Traditional Chinese Medicine X both showed obvious decreases, with the levels being 1.467 ± 0.545 g and 1.309 ± 0.603 g respectively, and the TGI levels being 56.35% and 61.05% respectively. The tumor inhibitory degrees were all statistically significant (P < 0.05), which were consistent with the positive control drugs, PD-1 antibody and cisplatin, at the same time. However, there was no obvious dose correlation in its tumor inhibitory effect. At the same time, in combination with PD-1 antibody or cisplatin treatment, Traditional Chinese Medicine X did not show an obvious synergistic effect on the proliferation inhibition of LLC xenografts. The tumor weights and TGI levels of the animals in each combination treatment group were the same as those in the single drug treatment. Among them, the tumor weights of the animals in the low and high dose of Traditional Chinese Medicine X combined with PD-1 antibody treatment groups were 1.356 ± 0.612 g and 1.116 ± 0.254 g respectively, and the TGI levels were 59.63% and 66.80% respectively. The tumor weights of the animals in the combination treatment group with cisplatin were 1.112 ± 0.544 g and 1.079 ± 0.253 g respectively, and the TGI levels were 66.89% and 67.88% respectively.

Claims

1. A traditional Chinese medicine composition for treating lung cancer, characterized in that: It is prepared from the following raw medicinal materials in the following weight ratios: 35 - 45 parts of Fagopyrum dibotrys, 25 - 35 parts of Marsdenia tenacissima, 15 - 25 parts of Ranunculus ternatus, 25 - 35 parts of Astragalus membranaceus, 15 - 25 parts of Glossy Privet Fruit, 25 - 35 parts of Caulis Spatholobi, 5 - 15 parts of Platycodon grandiflorum.

2. The traditional Chinese medicine composition for treating lung cancer according to claim 1, characterized in that: It is prepared from the following raw medicinal materials in the following weight ratios: 40 parts of Fagopyrum dibotrys, 30 parts of Marsdenia tenacissima, 20 parts of Ranunculus ternatus, 30 parts of Astragalus membranaceus, 20 parts of Glossy Privet Fruit, 30 parts of Caulis Spatholobi, 10 parts of Platycodon grandiflorum; the Glossy Privet Fruit is processed Glossy Privet Fruit.

3. The traditional Chinese medicine composition for treating lung cancer according to claim 1 or 2, characterized in that: It uses the raw medicinal powder of the above - mentioned raw medicinal materials, water or organic solvent extract as the active ingredient, and adds pharmaceutically acceptable excipients or auxiliary components to prepare common oral preparations in pharmacy.

4. The traditional Chinese medicine composition for treating lung cancer according to claim 3, characterized in that: The oral preparations are extractive agents, pills, dripping pills, granules, tablets, powders, capsules, tea preparations, oral liquids, syrups.

5. A method for preparing the traditional Chinese medicine composition for treating lung cancer according to any one of claims 1 - 4, characterized in that: It includes the following steps: a. Weigh the raw medicinal materials in the respective weight ratios. b. Directly powder the raw medicinal materials, extract them with water or organic solvents, and add pharmaceutically acceptable excipients or auxiliary components to prepare common oral preparations in pharmacy.

6. Use of the traditional Chinese medicine composition according to any one of claims 1 - 4 in the preparation of a drug for treating lung cancer.

7. The use according to claim 6, characterized in that: The lung cancer is non - small cell lung cancer.

8. The use according to claim 6, characterized in that: The lung cancer is lung cancer of the type of mutual binding of phlegm and stasis, deficiency of both qi and yin.

9. The use according to claim 6, characterized in that: The drug has the effects of shrinking tumors, inhibiting tumor growth, preventing tumor spread, and preventing tumor recurrence.

10. Use of the traditional Chinese medicine composition according to any one of claims 1 - 4 in the preparation of a drug with the effects of strengthening healthy qi and tonifying deficiency, dissipating swelling and resolving nodules, and tonifying the kidney and strengthening the spleen.

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