A traditional Chinese medicine composition for treating phlegm and qi mutual knotting type esophageal cancer and recurrence, and a preparation method and application thereof

Abstract

The present application mainly relates to a kind of traditional Chinese medicine composition for treating phlegm and qi mutual knotting type esophageal cancer and recurrence and preparation method and application thereof, belong to the field of biological medicine technology.The traditional Chinese medicine composition includes 3-90 parts of Huangqi, 1-60 parts of Donglingcao, 1-30 parts of Dangshen, 1-30 parts of Shengbaishu, 1-30 parts of Fuling, 1-30 parts of Chenpi, 1-30 parts of Chaihu, 1-30 parts of Weilingxian and 1-30 parts of Zhigancao.The above-mentioned raw materials are extracted by water to obtain the traditional Chinese medicine composition of the present application.The traditional Chinese medicine composition can correct the phlegm and qi mutual knotting symptoms of phlegm and qi mutual knotting type esophageal cancer patients, effectively inhibit the growth and recurrence of tumor, and has good safety.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to a traditional Chinese medicine composition for treating phlegm and qi mutual knotting type esophageal cancer and recurrence as well as a preparation method and application thereof. BACKGROUND

[0002] Esophageal cancer is a common digestive tract malignant tumor with high morbidity and mortality, which not only causes a serious disease burden, but also seriously threatens the health of residents. According to the histopathological characteristics, esophageal cancer can be divided into esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC). Nearly 75% of ESCC patients have developed into advanced or metastatic advanced stage. Although the comprehensive treatment scheme mainly based on surgery is constantly improved, the disease-free survival (DFS) and overall survival (OS) of esophageal cancer patients are both prolonged, however, after radical surgery for esophageal cancer, the quality of life of patients is poor, and the prognosis is also very unsatisfactory, and the 5-year survival rate is only between 15% and 20%. Recurrence and metastasis is an important factor affecting the long-term survival of patients. Regional recurrence and / or distant metastasis after surgery is the main mode of recurrence and metastasis of ESCC, especially regional recurrence and metastasis. Once the postoperative recurrence occurs, it indicates poor prognosis, and most patients die within 1 year. At present, the main salvage treatment for postoperative recurrence patients in clinical practice includes surgery, radiotherapy, chemotherapy and combined radiotherapy and chemotherapy, but the curative effects are not ideal and the side effects during treatment are large. Due to the lack of sufficient evidence-based medicine, there is no uniform standard for the radiotherapy target area, radiotherapy dose and combined chemotherapy mode of postoperative recurrence patients. Therefore, exploring new directions for the treatment of ESCC from various aspects and providing safer and more effective treatment strategies and drugs are technical problems to be solved in the field. SUMMARY

[0003] In view of this, the purpose of the present application is to provide a traditional Chinese medicine composition for treating phlegm and qi mutual knotting type esophageal cancer and recurrence, which can not only correct the symptoms of phlegm and qi mutual knotting in esophageal cancer patients, but also effectively inhibit the growth and recurrence of tumors, and has good safety.

[0004] The present application provides a traditional Chinese medicine composition for treating phlegm and qi mutual knotting type esophageal cancer and recurrence, which comprises the following raw materials by weight: 3-90 parts of Radix Astragali, 1-60 parts of Rabdosia rubescens, 1-30 parts of Radix Codonopsis, 1-30 parts of Radix Paeoniae Alba, 1-30 parts of Poria cocos, 1-30 parts of Pericarpium Citri Reticulatae, 1-30 parts of Bupleurum chinense, 1-30 parts of Cynanchum kousso, and 1-30 parts of Radix Glycyrrhizae.

[0005] Preferably, the traditional Chinese medicine composition for treating phlegm and qi mutual stagnation type esophageal cancer and recurrence comprises the following raw materials in parts by weight: 10-30 parts of radix astragali, 10-30 parts of rabdosia japonica, 3-9 parts of radix codonopsis, 3-9 parts of raw atractylodes, 3-9 parts of poria cocos, 3-9 parts of pericarpium citri reticulatae, 3-9 parts of radix bupleuri, 3-9 parts of radix clematidis, and 3-9 parts of prepared licorice.

[0006] The application further provides a preparation method of the traditional Chinese medicine composition, comprising the following steps: mixing and decocting all raw materials with water to extract, and collecting the decoction.

[0007] Preferably, the decocting and extracting is performed for 2 or more times.

[0008] Preferably, the decocting and extracting is performed for 2 or more times.

[0009] Preferably, the decocting and extracting is performed for 2 or more times.

[0010] Preferably, the preparation method further comprises a concentrating step; and the concentrating is concentrated to a relative density of 1.05-1.10.

[0011] Preferably, the preparation method further comprises a drying and granulating step; and the drying is dried to a moisture content of less than or equal to 6%.

[0012] The application further provides application of the traditional Chinese medicine composition or the preparation method in preparation of a medicine for treating esophageal cancer or recurrence of esophageal cancer.

[0013] Preferably, the esophageal cancer is phlegm and qi mutual stagnation type esophageal cancer.

[0014] The application has the following beneficial effects:

[0015] The application provides a traditional Chinese medicine composition for treating esophageal cancer. It has been found through experiments that, after an ESCC patient takes the traditional Chinese medicine composition of the application after surgery, the symptoms of phlegm and qi mutual stagnation in the body are obviously corrected. It has been found through mouse in-vivo experiments that the traditional Chinese medicine composition of the application can significantly inhibit the malignant growth of ESCC cell subcutaneous transplanted tumors, and compared with the first-line chemotherapy drug cisplatin, it can better inhibit the growth of ESCC recurrence tumors, and the effect has a significant difference. Meanwhile, biochemical examination research shows that the composition of the application is safe for internal use, and no obvious toxic and side effects are found. It shows that the traditional Chinese medicine composition of the application can effectively correct the symptoms of phlegm and qi mutual stagnation of esophageal cancer patients, and significantly inhibit the growth and recurrence of esophageal cancer tumors, and has good safety. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 The left side of the figure is a clinical test flowchart, and the right side of the figure is an animal in-vivo test flowchart.

[0017] Figure 2 Figure 1 is a tongue image feature map of preoperative and postoperative patients with ESCC; wherein, A is a representative tongue image of phlegm and qi mutual knotting type; B is a representative tongue image of blood stasis obstruction type; C is a representative tongue image of yin deficiency and heat knotting type; D is a representative tongue image of qi deficiency and yang deficiency type; E is a distribution map of each symptom of ESCC patients; F is a representative tongue image of preoperative (left) and postoperative (right) of phlegm and qi mutual knotting type patients.

[0018] Figure 3 Figure 1 is a tongue image feature map of preoperative and postoperative patients with ESCC; wherein, A is a representative tongue image of phlegm and qi mutual knotting type; B is a representative tongue image of blood stasis obstruction type; C is a representative tongue image of yin deficiency and heat knotting type; D is a representative tongue image of qi deficiency and yang deficiency type; E is a distribution map of each symptom of ESCC patients; F is a representative tongue image of preoperative (left) and postoperative (right) of phlegm and qi mutual knotting type patients.

[0019] Figure 4 Figure 1 is a tongue image feature map of preoperative and postoperative patients with ESCC; wherein, A is a representative tongue image of phlegm and qi mutual knotting type; B is a representative tongue image of blood stasis obstruction type; C is a representative tongue image of yin deficiency and heat knotting type; D is a representative tongue image of qi deficiency and yang deficiency type; E is a distribution map of each symptom of ESCC patients; F is a representative tongue image of preoperative (left) and postoperative (right) of phlegm and qi mutual knotting type patients.

[0020] Figure 5 Figure 1 is a tongue image feature map of preoperative and postoperative patients with ESCC; wherein, A is a representative tongue image of phlegm and qi mutual knotting type; B is a representative tongue image of blood stasis obstruction type; C is a representative tongue image of yin deficiency and heat knotting type; D is a representative tongue image of qi deficiency and yang deficiency type; E is a distribution map of each symptom of ESCC patients; F is a representative tongue image of preoperative (left) and postoperative (right) of phlegm and qi mutual knotting type patients.

[0021] Figure 6 Figure 1 is a tongue image feature map of preoperative and postoperative patients with ESCC; wherein, A is a representative tongue image of phlegm and qi mutual knotting type; B is a representative tongue image of blood stasis obstruction type; C is a representative tongue image of yin deficiency and heat knotting type; D is a representative tongue image of qi deficiency and yang deficiency type; E is a distribution map of each symptom of ESCC patients; F is a representative tongue image of preoperative (left) and postoperative (right) of phlegm and qi mutual knotting type patients. DETAILED DESCRIPTION

[0022] The application provides a traditional Chinese medicine composition for treating phlegm and qi mutual knotting type esophageal cancer and recurrence, which comprises the following raw materials in parts by weight: 3-90 parts of Astragalus membranaceus, 1-60 parts of Rabdosia rubescens, 1-30 parts of Codonopsis pilosula, 1-30 parts of raw Atractylodes macrocephala, 1-30 parts of Poria cocos, 1-30 parts of Pericarpium Citri Reticulatae, 1-30 parts of Bupleurum chinense, 1-30 parts of Clematis chinensis and 1-30 parts of prepared licorice; preferably, the composition comprises the following raw materials in parts by weight: 10-30 parts of Astragalus membranaceus, 10-30 parts of Rabdosia rubescens, 3-9 parts of Codonopsis pilosula, 3-9 parts of raw Atractylodes macrocephala, 3-9 parts of Poria cocos, 3-9 parts of Pericarpium Citri Reticulatae, 3-9 parts of Bupleurum chinense, 3-9 parts of Clematis chinensis and 3-9 parts of prepared licorice; more preferably, the composition comprises the following raw materials in parts by weight: 15-25 parts of Astragalus membranaceus, 15-25 parts of Rabdosia rubescens, 5-7 parts of Codonopsis pilosula, 5-7 parts of raw Atractylodes macrocephala, 5-7 parts of Poria cocos, 5-7 parts of Pericarpium Citri Reticulatae, 5-7 parts of Bupleurum chinense, 5-7 parts of Clematis chinensis and 5-7 parts of prepared licorice; further preferably, the composition comprises the following raw materials in parts by weight: 20 parts of Astragalus membranaceus, 20 parts of Rabdosia rubescens, 6 parts of Codonopsis pilosula, 6 parts of raw Atractylodes macrocephala, 6 parts of Poria cocos, 6 parts of Pericarpium Citri Reticulatae, 6 parts of Bupleurum chinense, 6 parts of Clematis chinensis and 6 parts of prepared licorice. The application does not have special limitation on the sources of the Astragalus membranaceus, Rabdosia rubescens, Codonopsis pilosula, raw Atractylodes macrocephala, Poria cocos, Pericarpium Citri Reticulatae, Bupleurum chinense, Clematis chinensis and prepared licorice, and the conventional commercially available products in the field can be adopted.

[0023] The traditional Chinese medicine composition of the application is composed of nine kinds of medicines, i.e. Astragalus membranaceus, Rabdosia rubescens, Codonopsis pilosula, raw Atractylodes macrocephala, Poria cocos, Pericarpium Citri Reticulatae, Bupleurum chinense, Clematis chinensis and prepared licorice. The compatibility of the medicines is as follows: the Astragalus membranaceus and Rabdosia rubescens are the monarch medicines, the Codonopsis pilosula, raw Atractylodes macrocephala, Poria cocos, Pericarpium Citri Reticulatae and Bupleurum chinense are the minister medicines, the Clematis chinensis is the assistant medicine, and the prepared licorice is the guide medicine. The Astragalus membranaceus is a long-term tonic medicine, has a sweet taste and a slightly warm nature, enters the spleen meridian, and can tonify the spleen and boost qi; the Rabdosia rubescens has a bitter taste and a slightly cold nature, can clear heat and resolve toxins, and can promote blood circulation and relieve pain; the Codonopsis pilosula has a sweet taste and a neutral nature, can invigorate the spleen and boost the lung, and can benefit qi and produce saliva; the raw Atractylodes macrocephala has a bitter taste and a warm nature, can invigorate the spleen and dry dampness; the Poria cocos has a sweet taste and a mild nature, can penetrate dampness and tonify the spleen; the Pericarpium Citri Reticulatae has an aromatic scent, can invigorate the spleen and harmonize the stomach, regulate qi and balance the center, and the Bupleurum chinense has a bitter taste and a slightly cold nature, can soothe the liver and relieve depression; the Clematis chinensis has a salty taste and a warm nature, can dispel wind and dampness, and can dredge the meridians; and the prepared licorice has a sweet and warm nature, and can regulate the center. The whole formula can boost qi, reduce adverse qi, invigorate the spleen and harmonize the stomach, and disperse knots and resolve masses, so that phlegm and saliva are consumed, adverse qi is reduced, and the center is invigorated, thereby helping to treat cancer.

[0024] The application further provides a preparation method of the traditional Chinese medicine composition, which comprises the following steps: mixing all the raw materials with water, decocting and extracting, and collecting the decoction.

[0025] In the present application, the number of decoction extraction is preferably ≥2 times, and in an embodiment, can be selected as 2 times, 3 times or 4 times. The time of each extraction is preferably 40-80 min after boiling, and in an embodiment, can be selected as 40 min, 50 min, 60 min, 65 min, 70 min or 80 min after boiling. The solid-liquid ratio of each extraction is preferably 1 g:(4-10) mL, and in an embodiment, can be selected as 1 g:4 mL, 1 g:5 mL, 1 g:6 mL, 1 g:7 mL, 1 g:8 mL, 1 g:9 mL or 1 g:10 mL, wherein the solid-liquid ratio refers to the ratio of raw material (crude drug) and water.

[0026] In the present application, the water extraction preferably further comprises a concentration step, and the concentration is preferably to a relative density of 1.05-1.10, and in an embodiment, can be selected as 1.05, 1.06, 1.07, 1.08, 1.09 or 1.10. The method of concentration is not particularly limited in the present application, and a conventional concentration method can be used. The concentration temperature is preferably not more than 100°C.

[0027] In the present application, the process preferably further comprises a drying and granulating step, and the drying is to a moisture content of ≤6%. The drying is preferably spray drying, and the inlet air temperature is preferably 135-165°C, the outlet air temperature is preferably 90-110°C, the pressure in the drying tower is preferably -0.1 to -0.6 kPa, and the wall temperature is preferably <45°C. During the drying process, the temperature and pressure parameters will fluctuate up and down, so as long as the temperature and pressure are controlled within the above ranges, the drying process can be carried out. After drying, the granules are preferably prepared by dry granulation, and the step of dry granulation is not particularly limited in the present application, and a conventional dry granulation step can be used.

[0028] The present application also provides the use of the traditional Chinese medicine composition or the preparation method in the preparation of a medicine for treating esophageal cancer or recurrence of esophageal cancer. The esophageal cancer is preferably phlegm and qi mutual knotting type esophageal cancer. The traditional Chinese medicine composition can effectively correct the phlegm and qi mutual knotting symptoms of patients with esophageal cancer, inhibit the growth and recurrence of esophageal cancer, and has good safety, and can effectively treat esophageal cancer.

[0029] In the present application, the medicine preferably further comprises a pharmaceutically acceptable carrier.

[0030] The technical solutions provided by the present application will be described in detail below in conjunction with the embodiments, but they should not be understood as limiting the scope of protection of the present application.

[0031] In the following examples, the conventional methods are used unless otherwise specified.

[0032] The materials, reagents and the like used in the following examples can be obtained commercially unless otherwise specified.

[0033] Example 1

[0034] A traditional Chinese medicine composition for treating phlegm and qi mutual stagnation type esophageal cancer and recurrence, which is composed of Radix Astragali 20g, Rabdosia rubescens 20g, Radix Codonopsis 6g, Radix Pseudostellariae 6g, Poria cocos 6g, Pericarpium Citri Reticulatae 6g, Radix Bupleuri 6g, Radix Clematidis 6g and Radix Glycyrrhizae 6g.

[0035] Preparation method: mix all the medicinal ingredients to obtain a mixture, add 8 times the amount of water (1g of mixture: 8mL of water) to the mixture, boil and decoct for 65min, pour out the decoction, add 6 times the amount of water (1g of mixture: 6mL of water), boil and decoct for 65min, pour out the decoction, and combine the two decoctions.

[0036] Concentrate the decoction, open the vacuum valve, vacuum degree: -0.03 to -0.07MPa, when the liquid surface of the material reaches 1 / 4 of the sight glass in the evaporator, close the feed valve; open the condenser and circulating cooling water, control the water pressure at 0.1 to 0.2MPa, slowly open the steam valve, increase the pressure to 0.03 to 0.06MPa, heat the material liquid in the concentrator to circulating boiling, control the temperature at 70 to 90℃, record the pressure and temperature every 1 hour during the concentration process, concentrate to obtain extract with a relative density of 1.08 (80℃). Spray dry, set the inlet air temperature at 135 to 165℃, the outlet air temperature at 90 to 110℃, the tower pressure at -0.1 to -0.6kPa, and the wall temperature <45℃. Collect the dry extract powder produced by spraying in time, and detect the moisture content, the moisture content of the dry extract powder ≤6%. Then dry granulate the dry extract powder, and collect the qualified granules, which are packaged by an automatic packaging machine.

[0037] Example 2

[0038] Composition: a traditional Chinese medicine composition for treating phlegm and qi mutual stagnation type esophageal cancer and recurrence, which is composed of Radix Astragali 10g, Rabdosia rubescens 30g, Radix Codonopsis 3g, Radix Pseudostellariae 9g, Poria cocos 3g, Pericarpium Citri Reticulatae 9g, Radix Bupleuri 3g, Radix Clematidis 9g and Radix Glycyrrhizae 3g.

[0039] Preparation method: mix all the medicinal ingredients to obtain a mixture, add 6 times the amount of water to the mixture, boil and decoct for 40min, pour out the decoction, add 4 times the amount of water, boil and decoct for 80min, pour out the decoction, and combine the two decoctions.

[0040] Concentrate the decoction, open the vacuum valve, vacuum degree: -0.03 to -0.07 MPa, when the liquid level of the material reaches 1 / 4 of the sight glass in the evaporator, close the feed valve; open the condenser and circulating cooling water, control the water pressure at 0.1 to 0.2 MPa, slowly open the steam valve, increase the temperature and pressure to 0.03 to 0.06 MPa, heat the liquid in the concentrator to circulating boiling, control the temperature at 70 to 90°C, record the pressure and temperature every 1 hour during the concentration process, concentrate to obtain extract with relative density 1.05 (75°C). Spray dry, set the inlet air temperature at 135 to 165°C, the outlet air temperature at 90 to 110°C, the tower pressure at -0.1 to -0.6 kPa, the wall temperature < 45°C. Collect the dry extract powder produced by the spray in time, and detect the moisture content, the moisture content of the dry extract powder ≤ 6%. Dry granulate the dry extract powder, and obtain qualified granules, which are packaged by an automatic packaging machine.

[0041] Example 3

[0042] Composition: a traditional Chinese medicine composition for treating phlegm and qi mutual accumulation type esophageal cancer and recurrence, which is composed of Huangqi 30 g, Donglingcao 10 g, Dangshen 9 g, Shengbaishu 3 g, Fuling 9 g, Chenpi 3 g, Chaihu 9 g, Weilingxian 3 g, and Zhigancao 9 g.

[0043] Preparation method: mix all the medicinal ingredients to obtain a mixture, add 10 times the amount of water to the mixture, boil and decoct for 80 min, pour out the decoction, and then add 8 times the amount of water, boil and decoct for 40 min, pour out the decoction, and combine the decoctions of the two times.

[0044] Concentrate the decoction, open the vacuum valve, vacuum degree: -0.03 to -0.07 MPa, when the liquid level of the material reaches 1 / 4 of the sight glass in the evaporator, close the feed valve; open the condenser and circulating cooling water, control the water pressure at 0.1 to 0.2 MPa, slowly open the steam valve, increase the temperature and pressure to 0.03 to 0.06 MPa, heat the liquid in the concentrator to circulating boiling, control the temperature at 70 to 90°C, record the pressure and temperature every 1 hour during the concentration process, concentrate to obtain extract with relative density 1.05 (75°C). Spray dry, set the inlet air temperature at 135 to 165°C, the outlet air temperature at 90 to 110°C, the tower pressure at -0.1 to -0.6 kPa, the wall temperature < 45°C. Collect the dry extract powder produced by the spray in time, and detect the moisture content, the moisture content of the dry extract powder ≤ 6%. Dry granulate the dry extract powder, and obtain qualified granules, which are packaged by an automatic packaging machine.

[0045] Example 4

[0046] Composition: a traditional Chinese medicine composition for treating phlegm and qi mutual accumulation type esophageal cancer and recurrence, which is composed of Huangqi 3 g, Donglingcao 1 g, Dangshen 1 g, Shengbaishu 1 g, Fuling 1 g, Chenpi 1 g, Chaihu 1 g, Weilingxian 1 g, and Zhigancao 1 g.

[0047] Preparation method: all the medicinal ingredients are mixed to form a mixture, 7 times the amount of water is added to the mixture, boiled and decocted for 50 min, the decoction is poured out and 5 times the amount of water is added, boiled and decocted for 70 min, the decoction is poured out, the decoction is poured out and 6 times the amount of water is added, boiled and decocted for 60 min, the decoction is poured out, the decoction is poured out and 4 times the amount of water is added, boiled and decocted for 50 min, the decoction is poured out, and the decoctions of the three times are combined.

[0048] The decoction is concentrated, the vacuum valve is opened, the vacuum degree is -0.03 to -0.07 MPa, when the liquid level of the material is 1 / 4 of the sight glass in the evaporator, the feeding valve is closed; the condenser and the circulating cooling water are opened, the water pressure is controlled at 0.1 to 0.2 MPa, the steam valve is slowly opened, the temperature and pressure are increased to 0.03 to 0.06 MPa, the material liquid in the concentrator is heated to circulating boiling, the temperature is controlled at 70 to 90°C, the pressure and temperature are recorded every 1 hour during the concentration process, the concentration is stopped until the relative density of the extract is 1.07 (78°C). Spray drying, set the inlet air temperature to 135 to 165°C, the outlet air temperature to 90 to 110°C, the tower pressure to -0.1 to -0.6 kPa, and the wall temperature to <45°C. The dry extract powder produced by spraying is collected in time and the moisture is detected, the moisture of the dry extract powder is ≤6%. The dry extract powder is then dry granulated to obtain qualified granules, which are packaged by an automatic packaging machine.

[0049] Example 5

[0050] Composition: a traditional Chinese medicine composition for treating esophageal cancer and recurrence of phlegm and qi mutual accumulation type, which is composed of Radix Astragali 90 g, Rabdosia rubescens 60 g, Radix Codonopsis 30 g, Rhizoma Bletillae 30 g, Poria cocos 30 g, Pericarpium Citri Reticulatae 30 g, Radix Bupleuri 30 g, Radix Clematidis 30 g, and Radix Glycyrrhizae 30 g.

[0051] Preparation method: all the medicinal ingredients are mixed to form a mixture, 9 times the amount of water is added to the mixture, boiled and decocted for 60 min, the decoction is poured out and 7 times the amount of water is added, boiled and decocted for 60 min, the decoction is poured out, the decoction is poured out and 6 times the amount of water is added, boiled and decocted for 60 min, the decoction is poured out, the decoction is poured out and 4 times the amount of water is added, boiled and decocted for 50 min, the decoction is poured out, and the decoctions of the four times are combined.

[0052] The decoction is concentrated, and the vacuum valve is opened to a vacuum degree of -0.03 to -0.07 MPa. When the liquid level reaches 1 / 4 of the sight glass in the evaporator, the feed valve is closed; the condenser and circulating cooling water are opened to control the water pressure at 0.1 to 0.2 MPa. The steam valve is slowly opened, and the temperature and pressure are increased to 0.03 to 0.06 MPa. The liquid in the concentrator is heated to circulating boiling, and the temperature is controlled at 70 to 90°C. During the concentration process, the pressure and temperature are recorded every hour, and the extract is concentrated to a relative density of 1.09 (82°C). For spray drying, the air inlet temperature is set to 135 to 165°C, the air outlet temperature to 90 to 110°C, the pressure in the tower to -0.1 to -0.6 kPa, and the wall temperature to <45°C. The dry paste powder produced by the spray is collected in time and tested for moisture. The moisture content of the dry paste powder is ≤6%. The dry paste powder is then dry granulated to obtain qualified granules, which are packaged using an automatic packaging machine.

[0053] Test Example 1

[0054] In order to verify the effect of the Chinese medicine composition of the present invention, clinical trials and in vivo animal experiments were carried out using the Chinese medicine composition prepared in Example 1 (named Qiling Jiedu Fang / Qiling Jiedu Granules). The experimental flow chart is shown in FIG. Figure 1 The specific test steps and results are as follows:

[0055] 1. Clinical Trials

[0056] Verification of the efficacy of Qiling Jiedu Decoction in treating patients with esophageal cancer after surgery with phlegm-qi syndrome

[0057] 1. Observe and summarize the tongue characteristics of ESCC patients before and after surgery

[0058] The efficacy of Traditional Chinese Medicine (TCM) has been repeatedly validated in clinical practice over thousands of years. However, TCM schools of thought are numerous, and their approaches to syndrome differentiation, treatment, and medication prescriptions for the same condition vary. This results in a lack of standardized criteria in actual clinical treatment, making it difficult to fully leverage TCM's strengths and achieve effective treatment outcomes. To address this, the present study, leveraging the modern TCM Tongue Diagnosis Image Analysis System (ZMT-1A), enrolled 89 patients with newly diagnosed ESCC. Tongue images were collected preoperatively, postoperatively, and after chemotherapy. Combined with clinical TCM judgment, these images were analyzed to identify and categorize ESCC syndromes. Tongue color, coating color, coating thickness, dryness, and dryness / greasyness were analyzed to identify and categorize ESCC syndromes.

[0059] Results: After analysis, ESCC patients were divided into the following four syndrome types: phlegm-qi syndrome, blood stasis syndrome, yin deficiency-heat syndrome, and qi deficiency-yang micro-syndrome. Figure 2 Among them, phlegm-qi syndrome accounted for 76% (68 people) ( Figure 2In addition, by comparing the tongue images of patients before and after surgery, it was found that with the surgical removal of the lesion, the TCM symptoms of patients with phlegm-qi syndrome did not improve significantly. Instead, phlegm-dampness worsened and Yin-Jin decreased. ( Figure 2 F in the figure). This indicates that the patient still has symptoms of phlegm and qi stagnation in his body and needs to be treated accordingly.

[0060] 2. Efficacy verification

[0061] 2.1 To investigate the efficacy of Qiling Jiedu Decoction, patients undergoing surgery for esophageal cancer with phlegm-qi syndrome were selected for validation. The inclusion, exclusion, termination, and diagnostic criteria were as follows:

[0062] (1) Inclusion criteria:

[0063] ① Patients with stage II or III primary esophageal squamous cell carcinoma confirmed by pathology or cytology and who have undergone the first postoperative chemotherapy;

[0064] ②Conform to the clinical syndrome differentiation standards of traditional Chinese medicine;

[0065] ③Age ≥18 and ≤85 years old, with good compliance;

[0066] ④ The patient received TP (paclitaxel + cisplatin) chemotherapy, the regimen description: paclitaxel (PTX) 210mg / m 2 Intravenous infusion of d1 + cisplatin (DDP) 30 mg / m 2 / d intravenous drip on days 1-3 (21 days as a cycle);

[0067] ⑤ After the standard TP chemotherapy regimen, the patient developed TCM syndrome of phlegm and qi;

[0068] ⑥ The subject has no major organ dysfunction, and blood routine, liver, kidney, and heart functions are normal;

[0069] ⑦ The informed consent form has been signed.

[0070] (2) Exclusion criteria:

[0071] ① Patients with abnormal thyroid function, diabetes, or hypertension;

[0072] ② Abnormal liver function (alanine aminotransferase, aspartate aminotransferase, total / direct / indirect bilirubin, bile acid, etc.) and / or abnormal renal function (urea, creatinine, uric acid, etc.);

[0073] ③ People with mental illness;

[0074] ④ Pregnant or breastfeeding women;

[0075] ⑤ History of other tumors within 5 years;

[0076] ⑥There is a family history of genetic diseases.

[0077] (3) Termination criteria

[0078] ① Severe adverse events, according to the doctor's judgment should stop the case of clinical trials;

[0079] ② Course of the disease, or the test appeared other impact on the observation of the disease, according to the doctor's judgment should stop the clinical trial, invalid case processing;

[0080] ③ Clinical trial plan implementation of important deviation, such as poor compliance, it is difficult to evaluate the drug effect;

[0081] ④ The subjects in the process of clinical trials, unwilling to continue the clinical trials, to the principal doctor put forward the requirements of the clinical trials.

[0082] (4) The standard of dropouts: subjects who have given informed consent and have been screened into the randomized trial, do not complete the treatment and observation period as stipulated in the protocol, are considered dropouts.

[0083] (5) Treatment of dropouts

[0084] ① When the subjects drop out, the researchers should take the door, appointment visits, telephone, letter and other ways, as far as possible to contact the subjects, ask the reason, record the last time of drug administration, complete the assessment project as far as possible.

[0085] ② Because of allergy or other adverse reactions, ineffective treatment and withdrawal of the test cases, the researchers should take appropriate treatment measures according to the actual situation of the subjects.

[0086] ③ Dropouts should be properly stored for the test data, both as a file, also for the full analysis and statistics required, dropouts do not need to be supplemented.

[0087] (6) Exclusion of test cases

[0088] ① The case selection does not meet the inclusion criteria, meet the exclusion criteria;

[0089] ② Has not used the test drug;

[0090] ③ After randomization, there is no data.

[0091] Before the statistical analysis of the data, the statistical personnel and the principal investigator discuss whether the case is excluded.

[0092] (7) Diagnostic criteria:

[0093] Western diagnostic criteria: The diagnostic criteria of esophageal squamous cell carcinoma were in accordance with the Guidelines for Diagnosis and Treatment of Esophageal Cancer (2020 edition) of the Chinese Society of Clinical Oncology (CSCO). The TNM staging of primary esophageal squamous cell carcinoma was in accordance with the 8th edition of the International Union Against Cancer (UICC) staging system.

[0094] TCM diagnostic criteria: The TCM syndrome types were in accordance with the Phlegm-Qi Mutual Knotting Type: reference was made to the Guidelines for Clinical Research on New Drugs of Traditional Chinese Medicine (2018 edition), Shanghai Guidelines for Diagnosis and Treatment of Traditional Chinese Medicine (2003 edition), and the Expert Consensus on Treatment of Esophageal Cancer in Combination with Traditional Chinese and Western Medicine (2021 edition).

[0095] Main symptoms: ① difficulty swallowing, ② chest fullness, ③ belching and vomiting, ④ red tongue, and ⑤ thin and greasy fur. Secondary symptoms: ① vomiting sputum, ② dry mouth and throat, and ③ wiry and slippery pulse. If the main symptoms meet three items, the secondary symptoms meet one item, or the main symptoms meet two items, and the secondary symptoms meet two items, it can be diagnosed. Based on years of clinical observation and experience in TCM department, two senior physicians judge the syndrome types.

[0096] 2.2 Management of trial drugs

[0097] (1) Name and specification of trial drugs

[0098] Patient treatment group drug: The traditional Chinese medicine composition prepared in Example 1 (named Qiling Jiedu Decoction) was prepared by Shanghai Wansicheng Pharmaceutical Co., Ltd. according to the prescription and preparation method of Example 1 of the present application.

[0099] Patient treatment group drug specification: 6 g / bag (2 times a day, 1 bag each time, oral administration).

[0100] (2) Distribution of trial drugs

[0101] Each research unit designates a trial drug administrator. The researcher screens eligible subjects, and after informed consent and writing of the research medical record, the trial drug administrator distributes the drugs according to the subject number, and records in the Clinical Trial Drug Usage Record Form.

[0102] (3) Trial drug inventory

[0103] At each visit, the trial drug administrator needs to record the amount of trial drug distributed, the amount taken by the subject, and the amount returned.

[0104] (4) Method of taking trial drugs

[0105] a. Patient treatment group: Qiling Jiedu Decoction (granules) 6 g / bag (2 times a day, 1 bag each time, oral administration), continuous administration for 21 days as one course of treatment, reexamination on the 22nd day, and treatment according to the current clinical routine method after the end of the clinical trial;

[0106] b. Patient control group: treated according to current clinical routine methods, without drug intervention.

[0107] (5) Test drug storage and recovery

[0108] a. The test drug cabinet is locked and stored at room temperature, keeping dry.

[0109] b. The remaining test drug recovered by the test drug administrator is stored separately and the remaining quantity is recorded on the "Clinical Test Drug Usage Record Form".

[0110] (6) Combined medication

[0111] a. Except for the test drug, the use of traditional Chinese medicine and western medicine for treating esophageal squamous cell carcinoma and other treatments related to esophageal squamous cell carcinoma during the observation period is prohibited.

[0112] b. The doctor should require the patient to bring all the drugs being taken to the visit to check the patient's combined medication. The drugs that must be continued to be taken for combined diseases, or other treatments must be recorded in the research medical record "Combined Medication Form" with the drug name (or other treatment name), dosage, usage frequency and time, etc. so as to be analyzed and reported when summarized.

[0113] 2.3. Verification results

[0114] After screening, a total of 60 patients with phlegm and qi mutual knotting type ESCC were included and randomly divided into two groups, a patient treatment group (experimental group) and a patient control group, each group of 30 people, taking the medicine according to the "2.2 item under (4) Test drug administration method" in this embodiment "I," and collecting the tongue image of the patient before and after taking the medicine.

[0115] Results: After one course of treatment with Qiling Jiedu Decoction, the postoperative patients with phlegm and qi mutual knotting type ESCC were verified based on the tongue diagnosis image analysis system (ZMT-1A) of traditional Chinese medicine. By comparing the tongue features of the postoperative ESCC patients before and after taking Qiling Jiedu Decoction, it was found that taking Qiling Jiedu Decoction could greatly correct the phlegm and qi mutual knotting symptoms (A in Figure 3 ) of the postoperative phlegm and qi mutual knotting type ESCC patients, while the phlegm and qi mutual knotting symptoms of the control group were not improved, and even significantly aggravated (B in Figure 3 ).

[0116] II. Animal in vivo test

[0117] 1. Effect of in vivo administration of the composition of the present application on the malignant growth and recurrence of esophageal cancer

[0118] 1.1 Test materials

[0119] (1) ESCC cell passage and culture

[0120] Human ESCC cell KYSE150 and murine ESCC cell mEC25 were purchased from the Cell Bank of the Chinese Academy of Sciences, both of which were identified by STR and well cultured in our laboratory. The culture medium of KYSE150 was RPMI 1640 medium containing 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin. The culture medium of mEC25 was high-glucose DMEM medium containing 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin. The cells were cultured at 37°C, 95% humidity and 5% CO2. When the cells were subcultured, 0.25% trypsin was used for digestion.

[0121] (2) Breeding of experimental animals

[0122] BALB / c-nude female mice (6 weeks old, weighing about 20 g) were purchased from Shanghai Slac Laboratory Animal Co., Ltd. C57BL / 6J female mice (6 weeks old, weighing about 20 g) were purchased from Shanghai Slac Laboratory Animal Co., Ltd. The breeding and experiments strictly followed the standard requirements of SPF level, and the animal experiments in this study were approved by the Ethics Committee of Shanghai University of Traditional Chinese Medicine (Approval No. PZSHUTCM2306250002).

[0123] (3) Preparation of drugs

[0124] The granules were prepared by the preparation method of Example 1 and stored in a-80°C refrigerator for standby use.

[0125] 1.2 Test scheme and results

[0126] 1.2.1 Anti-tumor effect and safety of Qiling Jiedu Formula

[0127] 1.2.1.1 Subcutaneous transplantation tumor model test of KYSE150 cells

[0128] (1) Test scheme: To investigate the anti-tumor effect of the composition of the present application (Qiling Jiedu Formula), a subcutaneous transplantation tumor model of esophageal cancer cells was constructed. The subcutaneous transplantation tumor model of KYSE150 cells was constructed as follows: after 7 days of adaptation, KYSE150 cells in the logarithmic growth phase were collected by centrifugation after digestion, and the cells were washed twice with PBS. A sufficient amount of cells was resuspended with serum-free medium, and KYSE150 cells (100 μL of cell suspension, about 2×10 6One) was inoculated subcutaneously in the left axillary of 26 nude mice. After the subcutaneous transplanted tumor grew stably, the groups were randomly divided again. The tumor-bearing placebo group (Vehicle, n=7) was given 200 μL of normal saline by gavage every day, the tumor-bearing low-dose administration group (Low dose, n=7) was given 200 μL of Qiling Jiedu granules normal saline solution (the dose was 1.82 g / kg, equivalent to the crude drug amount of 18.655 g / kg) by gavage every day, the tumor-bearing high-dose administration group (High dose, n=7) was given 200 μL of Qiling Jiedu granules normal saline solution (the dose was 5.46 g / kg, equivalent to the crude drug amount of 55.965 g / kg) by gavage every day, and the tumor-bearing cisplatin control group (CDDP, n=5) was given 100 μL of cisplatin normal saline solution (the dose was 5 mg / kg) by intraperitoneal injection every two days.

[0129] (2) Detection method and results:

[0130] ① The first day after KYSE150 cells were inoculated, the size of the transplanted tumor was measured at regular time intervals, the change of tumor volume with time was recorded, and the body weight and diet were recorded. The tumor volume: long diameter a, short diameter b, the tumor volume (TV) formula was calculated as: tumor volume (TV) = 1 / 2 × a × b 2 After about 2 weeks of administration, the mice were sacrificed, and the samples were taken, weighed and photographed.

[0131] The results are shown in A and B of Figure 4 , and the in vivo administration of Qiling Jiedu prescription can significantly inhibit the malignant growth of KYSE150 cell subcutaneous transplanted tumor.

[0132] ② Detect the phenotype index of subcutaneous tumor in different groups of mice: take the tumor tissues of Vehicle and High dose groups, and detect the protein expression of PCNA and CyclinD1 in the tissues by Western blot method.

[0133] The results are shown in E of Figure 4 , and it was found by Western blot detection that oral gavage of Qiling Jiedu prescription can significantly down-regulate the expression of cell proliferation index PCNA and cell cycle index Cyclin D1 in ESCC subcutaneous transplanted tumor.

[0134] 1.2.1.2 mEC25 cell subcutaneous transplanted tumor model test

[0135] (1) Test scheme: after C57BL / 6J female mice were adapted for 7 days, the logarithmic growth period of mEC25 cells was taken, digested and centrifuged to collect, and the cells were washed twice with PBS. Resuspend enough amount of cells with serum-free medium and matrix glue at a ratio of 1:1, and inoculate mEC25 cells (100 μL of cell suspension, about 6×10 6One) inoculated in the left axillary subcutaneous of 20 mice. After the subcutaneous tumor grew stably, re-randomized into tumor-bearing placebo group (Vehicle, n = 5), tumor-bearing low-dose administration group (Low dose, n = 5), tumor-bearing high-dose administration group (High dose, n = 5) and tumor-bearing cisplatin control group (CDDP, n = 5), each group was administered in the same way as the test scheme under item (1) in "1.2.1.1".

[0136] (2) Detection method and result:

[0137] Inoculate mEC25 cells as the first day, measure the size of the transplanted tumor volume regularly, record the change of the transplanted tumor volume with time, and record the body weight and diet. Tumor volume: long diameter a, short diameter b, the formula for calculating tumor volume (TV) is: Tumor volume (TV) = 1 / 2 × a × b 2 After about 2 weeks of administration, we will kill the mice, take the samples, weigh and take photos.

[0138] The results are shown as C and D in Figure 4 The in vivo administration of Qiling Jiedu Formula can significantly inhibit the malignant growth of mEC25 cell subcutaneous transplanted tumor.

[0139] The above results show that the in vivo administration of Qiling Jiedu Formula can significantly inhibit the malignant growth of ESCC cell subcutaneous transplanted tumor.

[0140] 1.2.1.3 Safety test

[0141] (1) Test scheme: Explore whether Qiling Jiedu Formula has obvious toxic side effects in vivo. Set up 6 groups: 2 groups without constructing transplanted tumor, including healthy control group (HC, 200 μL of normal saline was given by gavage every day), toxic side group (TS, 200 μL of Qiling Jiedu Formula granules normal saline solution was given by gavage every day, the dose was 5.46 g / kg), and 4 groups of subcutaneous transplanted tumor model constructed by KYSE150 cells, including Vehicle group (200 μL of normal saline was given by gavage every day), Low dose group (200 μL of Qiling Jiedu Formula granules normal saline solution was given by gavage every day, the dose was 1.82 g / kg), High dose group (200 μL of Qiling Jiedu Formula granules normal saline solution was given by gavage every day, the dose was 5.46 g / kg) and CDDP group (100 μL of cisplatin normal saline solution was given by intraperitoneal injection every two days, the dose was 5 mg / kg).

[0142] (2) Detection method and result:

[0143] ① Inoculate KYSE150 cells as the first day, measure the body weight and diet of mice regularly, kill the mice after about 2 weeks of administration, take the samples, weigh and take photos.

[0144] The results are as follows Figure 5 As shown, there was no significant difference in body weight between the TS group and the HC group ( Figure 5 A), there was no statistical difference in dietary intake ( Figure 5 Compared with the Vehicle group, the body weight of the mice in the CDDP group was significantly reduced ( Figure 5 In addition, the organs of the mice were weighed. The results showed that compared with the HC group, the kidney weight of the mice in the TS group decreased, while the liver and spleen weights showed no significant difference ( Figure 5 Compared with the Vehicle group, the weight of each organ in the CDDP group was significantly reduced ( Figure 5 CE in ).

[0145] ② The blood routine of the HC group, TS group, Vehicle group and CDDP group was tested, and the blood routine indicators of white blood cells (WBC), platelets (PLT) and red blood cells (RBC) were compared. Blood routine test method: The mice were fasted for 12 hours and deprived of water for 1 hour before blood sample collection. 1.0-1.5mL of blood was collected from the eyeball, and 20μL of blood was taken and injected into an anticoagulant tube containing 2% sodium heparin. The values ​​of white blood cells (WBC), platelets (PLT) and red blood cells (RBC) of the mice were detected using a Bowlinman (BM860) automatic analyzer.

[0146] The results are as follows Figure 5 As shown in the FH, it can be seen that compared with the Vehicle group, the number of PLTs in the CDDP group was significantly reduced, with no significant effect on red blood cells and white blood cells. Compared with the HC group, oral administration of the Qiling Jiedu Recipe significantly increased the platelet count in the mice's blood, without significantly affecting red blood cells or white blood cells.

[0147] In summary, oral administration of Qiling Jiedu Granules had no obvious toxic side effects, while intraperitoneal injection of CDDP caused significant changes in the body weight and organs of mice.

[0148] 1.2.2 Effect and safety of Qiling Jiedu prescription on tumor recurrence

[0149] (1) Test scheme: In order to investigate the effect of Qiling Jiedu recipe on the recurrence of esophageal cancer, human cell line KYSE150 was selected to construct a mouse subcutaneous tumor model. The subcutaneous tumor model of KYSE150 cells was constructed as follows: after 7 days of adaptation, KYSE150 cells in logarithmic growth phase were collected by centrifugation after digestion, and the cells were washed twice with PBS. Resuspend the cells with enough amount of serum-free medium, inoculate KYSE150 cells (100 μL of cell suspension, about 2 x 10 6 When the tumor volume grows to about 45mm 3 , give cisplatin by intraperitoneal injection, and stop cisplatin administration when the tumor shrinks to the size of a rice grain. Then wait for the tumor to grow again, and then randomly divide the groups again. Recurrence placebo group (RV, n = 7), recurrence low-dose administration group (RL, n = 7), recurrence high-dose administration group (RH, n = 7) and recurrence cisplatin control group (RC, n = 7). The RV group was given 200 μL of normal saline by gavage every day, the RL group was given 200 μL of Qiling Jiedu recipe granules normal saline solution (dose 1.82 g / kg) by gavage every day, the RH group was given 200 μL of Qiling Jiedu recipe granules normal saline solution (dose 5.46 g / kg) by gavage every day, and the RC group was given 100 μL of cisplatin normal saline solution (dose 5 mg / kg) by intraperitoneal injection every two days.

[0150] (2) Detection method and results:

[0151] Measure the size of the transplanted tumor at regular intervals, record the change of the transplanted tumor with time, and record the body weight. Tumor volume: long diameter a, short diameter b, the formula for calculating the tumor volume (TV) is: Tumor volume (TV) = 1 / 2 x a x b 2 After 24 days of administration, the mice were sacrificed, and the subcutaneous tumor, kidney, liver and other tissues of the mice were weighed and photographed.

[0152] ① Inhibition effect on tumor recurrence

[0153] The results are shown in A of Figure 6 , and the administration of Qiling Jiedu recipe in vivo can significantly inhibit the malignant growth of esophageal cancer recurrence.

[0154] ② Safety results

[0155] Compared with the recurrence placebo group, there was no significant difference in body weight of mice in the recurrence low-dose administration group and the recurrence high-dose administration group, while compared with the recurrence placebo group, the body weight of mice in the recurrence cisplatin group was found to be significantly reduced Figure 6B) in the results. In addition, we also weighed the organs of mice, and found that there was no significant difference in the weight of liver, kidney, spleen and heart of mice in the relapse low-dose administration group and the relapse high-dose administration group compared with the relapse placebo group; and the weight of liver and spleen of mice in the relapse cisplatin group was significantly reduced compared with the relapse placebo group (C and D in the results). Figure 6

[0156] In summary, oral gavage of Qiling Jiedu granules has no obvious toxic side effects, while after intraperitoneal injection of CDDP, the body weight and organs of mice are significantly changed.

[0157] The above only describes the preferred embodiments of the present application, and it should be noted that for those skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, and these improvements and refinements should also be considered as the protection scope of the present application.​

Claims

1. A Chinese medicine composition for treating esophageal cancer of phlegm-qi type and its recurrence, characterized in that: The invention is composed of the following raw materials in parts by weight: 10-30 parts of astragalus, 10-30 parts of rubescens, 3-9 parts of codonopsis, 3-9 parts of raw atractylodes, 3-9 parts of poria, 3-9 parts of tangerine peel, 3-9 parts of bupleurum, 3-9 parts of clematis and 3-9 parts of roasted liquorice.

2. The method for preparing the Chinese medicine composition according to claim 1, characterized in that: The following steps are involved: Mix all the raw materials with water, boil and extract, and collect the decoction.

3. The preparation method according to claim 2, characterized in that The decoction and extraction are performed ≥2 times.

4. The preparation method according to claim 3, characterized in that The extraction time for each decoction is 40 to 80 minutes after boiling.

5. The preparation method according to claim 3, characterized in that The material-liquid ratio of the decoction extraction is 1g: (4-10)mL.

6. The preparation method according to claim 2, characterized in that The method further comprises a concentration step; the concentration step is to concentrate the mixture to a relative density of 1.05 to 1.

10.

7. The preparation method according to claim 6, characterized in that The method also includes a drying and granulation step; the drying step is drying to a moisture content of ≤6%.

8. Use of the traditional Chinese medicine composition according to claim 1 or the preparation method according to any one of claims 2 to 7 in preparing a drug for treating esophageal cancer or recurrence of esophageal cancer.

9. The use according to claim 8, characterized in that The esophageal cancer is a phlegm-qi type esophageal cancer.

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