Traditional Chinese medicine composition for treating esophageal cancer with syndrome of intermingled phlegm and qi and recurrence as well as preparation method and application of traditional Chinese medicine composition

Abstract

The invention mainly relates to a traditional Chinese medicine composition for treating phlegm-qi stagnation type esophageal cancer and relapse and a preparation method and application thereof, and belongs to the technical field of biological medicine. The traditional Chinese medicine composition comprises 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 koidz, 1-30 parts of poria cocos, 1-30 parts of pericarpium citri reticulatae, 1-30 parts of radix bupleuri, 1-30 parts of radix clematidis and 1-30 parts of honey-fried licorice root. The raw materials are subjected to water extraction to obtain the traditional Chinese medicine composition. The traditional Chinese medicine composition disclosed by the invention can be used for correcting the symptom of intermingled phlegm and qi of a patient with the intermingled phlegm and qi type esophageal cancer and effectively inhibiting growth and relapse of tumors, and has good safety at the same time.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and in particular relates to a traditional Chinese medicine composition for treating esophageal cancer of phlegm-qi entanglement type and recurrence thereof, and a preparation method and application thereof. Background Art

[0002] Esophageal cancer is a common digestive tract malignancy with high morbidity and mortality, which not only causes a serious disease burden, but also seriously threatens the health of residents. According to 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 stage or with metastasis. Although the comprehensive treatment plan mainly based on surgery has been continuously improved, the disease-free survival (DFS) and overall survival (OS) of esophageal cancer patients have been prolonged. However, after radical surgery for esophageal cancer, the quality of life of patients is poor, and the prognosis is also not ideal, with a 5-year survival rate of only 15%-20%. Recurrence and metastasis are important factors affecting the long-term survival of patients. Regional recurrence and metastasis and / or distant metastasis after ESCC surgery are the main recurrence and metastasis patterns, especially regional recurrence and metastasis. Once a patient has a postoperative recurrence, it indicates a poor prognosis, and most patients die within 1 year. At present, the main salvage treatments for patients with postoperative recurrence include surgery, radiotherapy, chemotherapy, and combined radiotherapy and chemotherapy, but the efficacy is not ideal and the side effects are large during the treatment. Due to the lack of sufficient evidence-based medicine, there is no unified standard for radiotherapy target area, radiotherapy dose, and combined chemotherapy for patients with postoperative recurrence. Therefore, exploring new directions for ESCC treatment from various angles and providing safer and more effective treatment strategies and therapeutic drugs are technical issues that need to be solved in this field. Summary of the invention

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

[0004] The present invention provides a traditional Chinese medicine composition for treating esophageal cancer of phlegm-qi stagnation type and its recurrence, comprising the following raw materials in parts by weight: 3-90 parts of astragalus, 1-60 parts of rubescens, 1-30 parts of codonopsis, 1-30 parts of raw atractylodes, 1-30 parts of poria, 1-30 parts of tangerine peel, 1-30 parts of bupleurum, 1-30 parts of clematis and 1-30 parts of roasted liquorice.

[0005] Preferably, the Chinese medicine composition for treating esophageal cancer of phlegm-qi accumulation type and its recurrence comprises 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 Atractylodes, 3-9 parts of Poria, 3-9 parts of Citrus reticulata, 3-9 parts of Bupleurum, 3-9 parts of Clematidis and 3-9 parts of Radix Glycyrrhizae.

[0006] The invention also provides a method for preparing the Chinese medicine composition, comprising the following steps: mixing all raw materials with water, boiling and extracting, and collecting the decoction.

[0007] Preferably, the decoction extraction is performed ≥2 times.

[0008] Preferably, the time for each decoction extraction is 40 to 80 minutes after boiling.

[0009] Preferably, the material-liquid ratio of the decoction extraction is 1 g: (4-10) mL.

[0010] Preferably, the preparation method further comprises a concentration step; the concentration step is to concentrate to a relative density of 1.05 to 1.10.

[0011] Preferably, the preparation method further comprises a drying and granulation step; the drying is performed until the moisture content is ≤6%.

[0012] The present invention also provides application of the traditional Chinese medicine composition or the preparation method in preparing medicine for treating esophageal cancer or recurrence of esophageal cancer.

[0013] Preferably, the esophageal cancer is esophageal cancer of the phlegm-qi intertwined type.

[0014] Beneficial effects of the present invention:

[0015] The present invention provides a Chinese medicine composition for treating esophageal cancer. It is found through experiments that after ESCC patients take the Chinese medicine composition of the present invention after surgery, the symptoms of phlegm and qi stagnation in the body are significantly corrected. In vivo experiments on mice have found that the in vivo administration of the Chinese medicine composition of the present invention can significantly inhibit the malignant growth of subcutaneous transplanted tumors of ESCC cells, and compared with the first-line chemotherapy drug cisplatin, it can better inhibit the growth of ESCC recurrent tumors, and the effect is significantly different. At the same time, biochemical examination studies have shown that the composition of the present invention is safe for oral administration, and no obvious toxic and side effects are observed. It shows that the Chinese medicine composition of the present invention can effectively correct the symptoms of phlegm and qi stagnation in patients with esophageal cancer, and significantly inhibit the growth and recurrence of esophageal cancer tumors, and has good safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The figure on the left is a flow chart of the clinical trial flow chart, and the figure on the right is a flow chart of the in vivo animal test flow chart.

[0017] Figure 2 The tongue image characteristics of ESCC patients before and after surgery; A is a representative tongue image of the phlegm-qi type; B is a representative tongue image of the blood stasis type; C is a representative tongue image of the yin deficiency and heat type; D is a representative tongue image of the qi deficiency and yang micro type; E is a distribution diagram of the proportion of various symptoms in ESCC patients; F is a representative tongue image of patients with phlegm-qi type before (left) and after (right) surgery.

[0018] Figure 3 Graph showing the effect of the Chinese medicine composition of the present invention on postoperative patients with ESCC of phlegm-qi binding type; wherein A is the tongue image of the experimental group before (left) and after (right) taking the Chinese medicine composition of the present invention; and B is the tongue image of the control group before (left) and after (right) not taking the Chinese medicine composition of the present invention.

[0019] Figure 4 The figures are the effect diagrams of the in vivo administration of the composition of the present invention on inhibiting the subcutaneous transplantation of ESCC cells; wherein A is a diagram showing the volume change of the subcutaneous transplantation of KYSE150 cells in each group; B is a diagram showing the pictures and weight change of the subcutaneous transplantation of KYSE150 cells in each group; C is a diagram showing the volume change of the subcutaneous transplantation of mEC25 cells in each group; D is a diagram showing the pictures and weight change of the subcutaneous transplantation of mEC25 cells in each group; E is a diagram showing the effect of the Chinese medicine composition of the present invention on the expression of PCNA and CyclinD1 in the subcutaneous transplantation of KYSE150 cells; *P<0.05, **P<0.01, ***P<0.001.

[0020] Figure 5 The figures are graphs showing the results of the safety test of the Chinese medicine composition of the present invention for in vivo administration; wherein, A is a graph showing the changes in the body weight of mice; B is a graph showing the changes in the amount of food consumed by mice; C is a graph showing the changes in the liver weight of mice; D is a graph showing the changes in the kidney weight of mice; E is a graph showing the changes in the spleen weight of mice; FH are graphs showing the results of routine blood tests of mice with subcutaneous tumors transplanted with KYSE150 cells, wherein F is red blood cells, G is white blood cells, and H is platelets; *P<0.05, **P<0.01, ***P<0.001, ns is P≥0.05.

[0021] Figure 6 The figures are test results of the inhibitory effect and safety of the Chinese medicine composition of the present invention on esophageal cancer recurrence; wherein A is the changes in tumor volume (left), tumor picture (upper right) and tumor weight (lower right) of mice with tumor recurrence; B is the changes in body weight of mice with tumor recurrence; C is the changes in liver weight of mice with tumor recurrence; D is the changes in kidney (left), spleen (middle) and heart (right) weight of mice with tumor recurrence; *P<0.05, **P<0.01, ***P<0.001, ns is P≥0.05. DETAILED DESCRIPTION

[0022] The present invention provides a Chinese medicine composition for treating esophageal cancer of phlegm-qi stagnation type and recurrence, comprising the following raw materials in parts by weight: 3-90 parts of astragalus, 1-60 parts of rubescens, 1-30 parts of codonopsis, 1-30 parts of raw atractylodes, 1-30 parts of tuckahoe, 1-30 parts of tangerine peel, 1-30 parts of bupleurum, 1-30 parts of clematis and 1-30 parts of roasted liquorice; preferably comprising 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 tuckahoe, 3-9 parts of tangerine peel, 3-9 parts of bupleurum, 3-9 parts of clematis and 3-9 parts of licorice; more preferably, the following raw materials in parts by weight: 15-25 parts of astragalus, 15-25 parts of winter grass, 5-7 parts of codonopsis, 5-7 parts of raw atractylodes, 5-7 parts of tuckahoe, 5-7 parts of tangerine peel, 5-7 parts of bupleurum, 5-7 parts of clematis and 5-7 parts of licorice; further preferably, the following raw materials in parts by weight: 20 parts of astragalus, 20 parts of winter grass, 6 parts of codonopsis, 6 parts of raw atractylodes, 6 parts of tuckahoe, 6 parts of tangerine peel, 6 parts of bupleurum, 6 parts of clematis and 6 parts of licorice. The present invention does not specifically limit the sources of the astragalus, winter grass, codonopsis, raw atractylodes, tuckahoe, tangerine peel, bupleurum, clematis and licorice, and conventional commercial products in the field can be used.

[0023] The Chinese medicine composition of the present invention is composed of nine herbs: Astragalus, Rubescens, Codonopsis, Atractylodes macrocephala, Poria, Tangerine peel, Bupleurum, Clematidis and Radix Glycyrrhizae. The monarch, minister, assistant and envoy compatibility is as follows: monarch: Astragalus, Rubescens; minister: Codonopsis, Atractylodes macrocephala, Poria, Tangerine peel, Bupleurum; assistant: Clematidis; envoy: Glycyrrhizae. Among them, Astragalus is the "leader of tonics", sweet in taste and slightly warm in nature, enters the spleen meridian, invigorates the spleen and replenishes qi, Rubescens is bitter in taste and slightly cold in nature, clears away heat and detoxifies, promotes blood circulation and relieves pain, and is the monarch drug; Codonopsis is sweet in taste and flat in nature, invigorates the spleen and replenishes the lungs, invigorates qi and produces body fluid, Atractylodes macrocephala is bitter in taste and warm in nature, invigorates the spleen and dries dampness, Poria is sweet in taste and light in nature, penetrates dampness and invigorates the spleen, Tangerine peel is aromatic, invigorates the spleen and smoothes the stomach, regulates qi and regulates the middle, Bupleurum is bitter in taste and slightly cold in nature, soothes the liver and relieves depression, and is the minister drug; Clematidis is salty in taste and warm in nature, removes rheumatism, dredges the meridians, and is the adjuvant drug; Glycyrrhizae is sweet and warm in nature, regulates the middle, and is the envoy drug. The whole formula works together to replenish Qi and reduce adverse effects, replenish Qi and harmonize the stomach, reduce swelling and disperse nodules, so that phlegm can be eliminated, adverse Qi can be calmed, and deficiency in the middle can be restored, so as to strengthen the body and treat cancer.

[0024] The invention also provides a method for preparing the Chinese medicine composition, comprising the following steps: mixing all raw materials with water, boiling and extracting, and collecting the decoction.

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

[0026] In the present invention, after the water extraction, it is preferred that a concentration step is further included, and the concentration is preferably concentrated to an extract with a relative density of 1.05 to 1.10. In one embodiment, the extract can be concentrated to a relative density of 1.05, 1.06, 1.07, 1.08, 1.09 or 1.10. The present invention does not specifically limit the concentration method, and a conventional concentration method in the art can be used. The preferred concentration temperature in the concentration process of the present invention does not exceed 100°C.

[0027] In the present invention, it is preferred that the step of drying and granulation is also included, and the drying is drying to a moisture content of ≤6%. The drying is preferably spray drying, and the inlet air temperature of the spray drying is preferably 135-165°C, the outlet air temperature is preferably 90-110°C, the pressure in the drying tower is preferably -0.1-0.6kPa, and the wall protection temperature is preferably <45°C. During the drying process, the temperature and pressure parameters will fluctuate up and down as the drying process proceeds, so it is sufficient to control the temperature and pressure within the above numerical range. After drying, preferably dry granulation is used to prepare the granules. The present invention does not specifically limit the step of dry granulation, and the conventional dry granulation steps in the art can be used for preparation.

[0028] The present invention also provides the use of the Chinese medicine composition or the preparation method in preparing a drug for treating esophageal cancer or esophageal cancer recurrence. The esophageal cancer is preferably esophageal cancer of the phlegm-qi stagnation type. The Chinese medicine composition of the present invention can effectively correct the phlegm-qi stagnation symptoms of esophageal cancer patients, and inhibit the growth and recurrence of esophageal cancer tumors, while having good safety and being able to effectively treat esophageal cancer.

[0029] In the present invention, the drug preferably further includes a pharmaceutically acceptable carrier.

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

[0031] In the following embodiments, unless otherwise specified, all of them are conventional methods.

[0032] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0033] Example 1

[0034] A traditional Chinese medicine composition for treating esophageal cancer of phlegm-qi stagnation type and its recurrence, comprising 20g of Astragalus, 20g of Rubus rubescens, 6g of Codonopsis pilosula, 6g of Atractylodes macrocephala, 6g of Poria, 6g of Tangerine peel, 6g of Bupleurum, 6g of Clematidis and 6g of Radix Glycyrrhizae Preparata.

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

[0036] The decoction is concentrated, and the vacuum valve is opened. The vacuum degree is -0.03~-0.07MPa. 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 0.1~0.2MPa, and the steam valve is slowly opened to increase the temperature and pressurize to 0.03~0.06MPa, so that the liquid in the concentrator is heated to circulating boiling, and the temperature is controlled at 70~90℃. During the concentration process, the pressure and temperature are recorded once every hour, and the extract with a relative density of 1.08 (80℃) is obtained. For spray drying, the air inlet temperature is set to 135~165℃, the air outlet temperature is set to 90~110℃, the pressure in the tower is -0.1~-0.6kPa, and the wall protection temperature is <45℃. The dry paste powder produced by the spray is collected in time and the moisture is tested. 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 by an automatic packaging machine.

[0037] Example 2

[0038] Prescription: A Chinese medicine composition for treating esophageal cancer of phlegm-qi stagnation type and its recurrence, consisting of 10g of Astragalus, 30g of Rubus rubescens, 3g of Codonopsis pilosula, 9g of Raw Atractylodes macrocephala, 3g of Poria, 9g of Tangerine peel, 3g of Bupleurum, 9g of Clematidis and 3g of Radix Glycyrrhizae.

[0039] Preparation method: Mix all the medicinal ingredients to make a mixture, add 6 times the amount of water to the mixture, boil it and simmer it for 40 minutes, pour out the decoction and add 4 times the amount of water again, boil it and simmer it for 80 minutes, pour out the decoction and combine the two decoctions.

[0040] The decoction is concentrated, and the vacuum valve is opened. The vacuum degree is -0.03~-0.07MPa. 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 0.1~0.2MPa, and the steam valve is slowly opened to increase the temperature and pressurize to 0.03~0.06MPa, so that the liquid in the concentrator is heated to circulating boiling, and the temperature is controlled at 70~90℃. During the concentration process, the pressure and temperature are recorded once every hour, and the extract with a relative density of 1.05 (75℃) is obtained. For spray drying, the air inlet temperature is set to 135~165℃, the air outlet temperature is set to 90~110℃, the pressure in the tower is -0.1~-0.6kPa, and the wall protection temperature is <45℃. The dry paste powder produced by the spray is collected in time and the moisture is tested. 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 by an automatic packaging machine.

[0041] Example 3

[0042] Prescription: A Chinese medicine composition for treating esophageal cancer of phlegm-qi stagnation type and its recurrence, consisting of 30g of Astragalus, 10g of Rubus rubescens, 9g of Codonopsis pilosula, 3g of Raw Atractylodes macrocephala, 9g of Poria, 3g of Tangerine peel, 9g of Bupleurum, 3g of Clematidis and 9g of Radix Glycyrrhizae Preparata.

[0043] Preparation method: Mix all the medicinal ingredients to make a mixture, add 10 times the amount of water to the mixture, boil it and simmer it for 80 minutes, pour out the decoction and add 8 times the amount of water again, boil it and simmer it for 40 minutes, pour out the decoction and combine the two decoctions.

[0044] The decoction is concentrated, and the vacuum valve is opened. The vacuum degree is -0.03~-0.07MPa. 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 0.1~0.2MPa, and the steam valve is slowly opened to increase the temperature and pressurize to 0.03~0.06MPa, so that the liquid in the concentrator is heated to circulating boiling, and the temperature is controlled at 70~90℃. During the concentration process, the pressure and temperature are recorded once every hour, and the extract with a relative density of 1.10 (85℃) is obtained. For spray drying, the air inlet temperature is set to 135~165℃, the air outlet temperature is set to 90~110℃, the pressure in the tower is -0.1~-0.6kPa, and the wall protection temperature is <45℃. The dry paste powder produced by the spray is collected in time and the moisture is tested. 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 by an automatic packaging machine.

[0045] Example 4

[0046] Prescription: A Chinese medicine composition for treating esophageal cancer of phlegm-qi stagnation type and its recurrence, consisting of 3g of Astragalus, 1g of Rabdosia rubescens, 1g of Codonopsis pilosula, 1g of Raw Atractylodes macrocephala, 1g of Poria, 1g of Tangerine peel, 1g of Bupleurum, 1g of Clematidis and 1g of Radix Glycyrrhizae.

[0047] Preparation method: Mix all the medicinal ingredients to make a mixture, add 7 times the amount of water to the mixture, boil it and simmer for 50 minutes, pour out the decoction and add 5 times the amount of water again, boil it and simmer for 70 minutes, pour out the decoction, pour out the decoction and add 6 times the amount of water again, boil it and simmer for 60 minutes, pour out the decoction, and combine the 3 decoctions.

[0048] The decoction is concentrated, and the vacuum valve is opened. The vacuum degree is -0.03~-0.07MPa. 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 0.1~0.2MPa, and the steam valve is slowly opened to increase the temperature and pressurize to 0.03~0.06MPa, so that the liquid in the concentrator is heated to circulating boiling, and the temperature is controlled at 70~90℃. During the concentration process, the pressure and temperature are recorded once every hour, and the extract with a relative density of 1.07 (78℃) is obtained. Spray drying, set the inlet temperature to 135~165℃, the outlet temperature to 90~110℃, the pressure in the tower to -0.1~-0.6kPa, and the wall protection temperature <45℃. The dry paste powder produced by the spray is collected in time and the moisture is tested. 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 by an automatic packaging machine.

[0049] Example 5

[0050] Prescription: A traditional Chinese medicine composition for treating esophageal cancer of phlegm-qi type and its recurrence, consisting of 90g of Astragalus, 60g of Rubus rubescens, 30g of Codonopsis pilosula, 30g of Raw Atractylodes macrocephala, 30g of Poria, 30g of Tangerine peel, 30g of Bupleurum, 30g of Clematidis and 30g of Radix Glycyrrhizae.

[0051] Preparation method: Mix all the medicinal ingredients to make a mixture, add 9 times the amount of water to the mixture, boil it and simmer for 60 minutes, pour out the decoction and add 7 times the amount of water again, boil it and simmer for 60 minutes, pour out the decoction, pour out the decoction and add 6 times the amount of water again, boil it and simmer for 60 minutes, pour out the decoction, pour out the decoction and add 4 times the amount of water again, boil it and simmer for 50 minutes, pour out the decoction, and combine the 4 decoctions.

[0052] The decoction is concentrated, and the vacuum valve is opened. The vacuum degree is -0.03~-0.07MPa. 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 0.1~0.2MPa, and the steam valve is slowly opened to increase the temperature and pressurize to 0.03~0.06MPa, so that the liquid in the concentrator is heated to circulating boiling, and the temperature is controlled at 70~90℃. During the concentration process, the pressure and temperature are recorded once every hour, and the extract with a relative density of 1.09 (82℃) is obtained. Spray drying, set the inlet temperature to 135~165℃, the outlet temperature to 90~110℃, the pressure in the tower to -0.1~-0.6kPa, and the wall temperature <45℃. The dry paste powder produced by the spray is collected in time and the moisture is tested. The moisture of the dry paste powder is ≤6%. The dry paste powder is then dry granulated to obtain qualified granules, which are packaged by 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 tests were carried out using the Chinese medicine composition prepared in Example 1 (named Qiling Jiedu Fang / Qiling Jiedu Granules). The test 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 has been repeatedly verified in clinical diagnosis and treatment for thousands of years. However, there are many theories of traditional Chinese medicine, and the same disease has different syndrome differentiation, treatment methods, and prescription drugs. This leads to no standard to rely on in the actual clinical treatment process, and it is difficult to ensure that the advantages of traditional Chinese medicine are used to achieve definite efficacy. To this end, the present invention relies on the modern instrument traditional Chinese medicine tongue diagnosis image analysis system (ZMT-1A), selects 89 patients with ESCC who have just been treated, collects tongue diagnosis images of patients before, after and after chemotherapy, and then combines clinical traditional Chinese medicine judgment for dialectical analysis. By analyzing the tongue color, coating color, thickness, moistness and greasiness of the tongue coating, the syndrome type of ESCC patients is judged and summarized.

[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, but instead worsened phlegm-dampness and decreased yin fluid ( 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 Fang, postoperative patients with esophageal cancer of the phlegm-qi type were selected for verification. The inclusion, exclusion, termination and diagnostic criteria were as follows:

[0062] (1) Inclusion criteria:

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

[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 underwent TP (paclitaxel + cisplatin) chemotherapy. The regimen description: paclitaxel (PTX) 210 mg / m 2 Intravenous infusion of d1 + cisplatin (DDP) 30 mg / m 2 / d intravenous drip d1-3, (21 days as a cycle);

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

[0068] ⑥ The subjects have no major organ dysfunction, and their 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 lactating women;

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

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

[0077] (3) Termination criteria

[0078] ① Those who have serious adverse events and the clinical trial of the case should be stopped according to the doctor's judgment;

[0079] ② If the condition worsens during the course of the disease, or other symptoms that affect the trial observation appear during the trial, the clinical trial should be stopped according to the doctor's judgment and treated as an invalid case;

[0080] ③ Important deviations occurred in the implementation of the clinical trial protocol, such as poor compliance, making it difficult to evaluate the drug effect;

[0081] ④ The subject is unwilling to continue the clinical trial during the clinical trial and requests to withdraw from the clinical trial to the attending physician.

[0082] (4) Dropout criteria: Subjects who have obtained informed consent and are eligible for randomized trials but fail to complete the course of treatment and observation period specified in this protocol for some reason will be considered dropouts.

[0083] (5) Treatment of dropout cases

[0084] ① When a subject drops out, the researcher should try to contact the subject by visiting, making appointments, calling, writing letters, etc., to ask the subject's reasons, record the last time he or she took the medication, and complete all the assessment items that can be completed.

[0085] ② For cases where the subjects withdraw from the trial due to allergic or other adverse reactions or ineffective treatment, the researchers should take appropriate treatment measures based on the actual situation of the subjects.

[0086] ③ The relevant test data of all dropped out cases should be properly preserved, both for archiving and for full analysis and statistics. No replacement is required for dropped out patients.

[0087] (6) Exclusion of test cases

[0088] ① The case selection did not meet the inclusion criteria but met the exclusion criteria;

[0089] ② Never used the experimental drug;

[0090] ③There is no data after randomization.

[0091] Before statistical analysis of the data, the statisticians and the principal investigator discussed and determined whether a case should be excluded.

[0092] (7) Diagnostic criteria:

[0093] Western medicine diagnostic criteria: The diagnostic criteria for esophageal squamous cell carcinoma are based on the "Guidelines for the Diagnosis and Treatment of Esophageal Cancer (2020 Edition)" of the Chinese Society of Clinical Oncology (CSCO). TNM staging of primary esophageal squamous cell carcinoma: According to the 8th edition of the International Union Against Cancer (UICC) staging system in 2017.

[0094] TCM diagnostic criteria: For patients whose TCM syndrome differentiation is consistent with the phlegm-qi syndrome: refer to the "Guidelines for Clinical Research of Syndrome-Based New Chinese Medicines (2018 Edition)", "Shanghai TCM Diagnosis and Treatment Routine (2003 Edition)" and "Expert Consensus on the Treatment of Esophageal Cancer with Integrated Traditional Chinese and Western Medicine (2021 Edition)".

[0095] Main symptoms: ① dysphagia, ② chest distension, ③ belching and vomiting, ④ red tongue with thin and greasy coating. Secondary symptoms: ① vomiting and sputum, ② dry mouth and throat, ③ stringy and slippery pulse. If the main symptoms meet three and the secondary symptoms meet one, or if the main symptoms meet two and the secondary symptoms meet two, the diagnosis can be confirmed. Combining the years of clinical observation and experience of the Department of Traditional Chinese Medicine, two senior doctors judged the syndrome types respectively.

[0096] 2.2 Trial Drug Management

[0097] (1) Name and specification of the trial drug

[0098] Drugs for the patient treatment group: The Chinese medicine composition prepared in Example 1 (named Qiling Jiedu Fang) was commissioned by Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine to be prepared by Shanghai Wanshicheng Pharmaceutical Co., Ltd. according to the formulation and preparation method of Example 1 of the present invention.

[0099] Drug specifications for the patient treatment group: 6g / packet (twice a day, 1 pack each time, orally).

[0100] (2) Distribution of trial drugs

[0101] Each research unit shall designate a trial drug manager. After the researcher screens qualified subjects and obtains informed consent and writes the research medical records, the trial drug manager shall distribute drugs case by case according to the subject serial number and register them in the "Clinical Trial Drug Use Record Form".

[0102] (3) Inventory of trial drugs

[0103] During each visit, the trial drug administrator must record the amount of trial drugs distributed, the amount taken by the subjects, and the amount returned.

[0104] (4) Method of taking the trial medication

[0105] a. Patient treatment group: Qiling Jiedu Fang (granules) 6g / packet (twice a day, 1 pack each time, orally), take the medicine for 21 consecutive days as a course of treatment, and re-examine on the 22nd day. After the end of the clinical trial, treatment will be based on current clinical routine methods;

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

[0107] (5) Storage and recovery of experimental drugs

[0108] a. The experimental drugs should be locked in the counter, stored at room temperature and kept dry.

[0109] b. The remaining trial drugs recovered by the trial drug administrator shall be stored separately, and the remaining quantity shall be recorded in the "Clinical Trial Drug Usage Record Form".

[0110] (6) Combined medication

[0111] a. Except for the experimental drugs, the use of Chinese and Western medicines for the treatment of esophageal squamous cell carcinoma and other treatments related to esophageal squamous cell carcinoma is prohibited during the observation period.

[0112] b. Doctors should ask patients to bring all the medications they are taking during the visit to check the patients' concomitant medications. The medications that must be continued for the concomitant diseases or other treatments must be recorded in the "Concomitant Medication Table" of the research medical record, including the name of the medication (or other treatment name), dosage, frequency of use and time, so that they can be analyzed and reported in the summary.

[0113] Verification results

[0114] After screening, a total of 60 ESCC patients with phlegm-qi intertwined type were included and randomly divided into 2 groups, a patient treatment group (experimental group) and a patient control group, with 30 patients in each group. The patients took the drugs according to "(4) Trial medication method under Item 2.2" of "I" of this embodiment, and the tongue images of the patients were collected before and after taking the drugs.

[0115] Results: After one course of treatment with Qiling Jiedu Fang, the selected patients with esophageal cancer with phlegm-qi intertwined after surgery were verified by using the TCM tongue diagnosis image analysis system (ZMT-1A). By comparing the tongue characteristics of ESCC patients before and after taking Qiling Jiedu Fang, it was found that taking Qiling Jiedu Fang could largely correct the phlegm-qi intertwined symptoms of ESCC patients with phlegm-qi intertwined after surgery ( Figure 3 A in the figure), while the symptoms of phlegm and qi accumulation in the control group did not improve and even became significantly worse ( Figure 3 B).

[0116] 2. Animal in vivo testing

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

[0118] 1.1 Test materials

[0119] (1) ESCC cell passaging and culture

[0120] Both human ESCC cells KYSE150 and mouse ESCC cells mEC25 were purchased from the cell bank of the Type Culture Committee, and were identified by STR. They were subcultured in this laboratory and were in good condition. The culture medium of KYSE150 was RPMI 1640 medium containing 10% fetal bovine serum, 100U / mL penicillin, and 100μg / mL streptomycin. The culture medium of mEC25 was high-glucose DMEM medium containing 10% fetal bovine serum, 100U / mL penicillin, and 100μg / mL streptomycin. The cells were placed at 37°C, 95% air humidity, and 5% CO 2 When cells were passaged, they were digested with 0.25% trypsin.

[0121] (2) Animal husbandry

[0122] BALB / c-nude female mice (6 weeks old, weighing about 20 g) were purchased from Shanghai Slack Experimental Animal Co., Ltd. C57BL / 6J female mice (6 weeks old, weighing about 20 g) were purchased from Shanghai Slack Experimental Animal Co., Ltd. The feeding and experiments strictly complied with the SPF grade standard requirements, and the animal experiments in this study were approved by the Ethics Committee of Shanghai University of Traditional Chinese Medicine (approval number: PZSHUTCM2306250002).

[0123] (3) Preparation of drugs

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

[0125] 1.2 Experimental plan and results

[0126] 1.2.1 Antitumor effect and safety of Qiling Jiedu prescription

[0127] 1.2.1.1 KYSE150 cell subcutaneous transplant tumor model test

[0128] (1) Experimental plan: In order to investigate the anti-tumor effect of the composition of the present invention (Qiling Jiedu Fang), a subcutaneous transplant tumor model of esophageal cancer cells was constructed. The subcutaneous transplant tumor model of KYSE150 cells was constructed as follows: After the nude mice were adapted for 7 days, KYSE150 cells in the logarithmic growth phase were obtained, digested and collected by centrifugation, and the cells were washed twice with PBS. Sufficient amount of cells were resuspended in serum-free medium, and KYSE150 cells (cell suspension 100 μL, cell number about 2×10 6) were inoculated subcutaneously in the left axilla of 26 nude mice. After the growth of subcutaneous transplanted tumors was stable, they were randomly divided into groups: the tumor-bearing placebo group (Vehicle, n=7) was given 200 μL of normal saline by gavage every day, the tumor-bearing low-dose group (Low dose, n=7) was given 200 μL of Qiling Jiedu Fang granules in normal saline solution (dose of 1.82 g / kg, equivalent to 18.655 g / kg of crude drug) by gavage every day, the tumor-bearing high-dose group (High dose, n=7) was given 200 μL of Qiling Jiedu Fang granules in normal saline solution (dose of 5.46 g / kg, equivalent to 55.965 g / kg of crude drug) by gavage every day, and the tumor-bearing cisplatin control group (CDDP, n=5) was given 100 μL of cisplatin in normal saline solution (dose of 5 mg / kg) by intraperitoneal injection every two days.

[0129] (2) Detection methods and results:

[0130] ① From the first day of KYSE150 cell inoculation, the size of the transplanted tumor was measured regularly, and the changes in tumor volume over time were recorded. The body weight and diet were also recorded. Tumor volume: long diameter a, short diameter b, the formula for calculating tumor volume (TV) is: Tumor volume (TV) = 1 / 2 × a × b 2 About 2 weeks after administration, we killed the mice, collected samples, weighed them, and took photos.

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

[0132] ② Detection of phenotypic indicators of subcutaneous tumors in mice of different groups: Tumor tissues from the Vehicle and High dose groups were taken, and the protein expressions of PCNA and CyclinD1 in the tissues were detected by Western blot.

[0133] The results are as follows Figure 4 As shown in Figure E, Western blot analysis revealed that oral administration of Qiling Jiedu Recipe could significantly down-regulate the expression of PCNA, a cell proliferation marker, and Cyclin D1, a cell cycle marker, in ESCC subcutaneous transplanted tumors.

[0134] 1.2.1.2 Subcutaneous transplantation tumor model experiment of mEC25 cells

[0135] (1) Experimental plan: After C57BL / 6J female mice were adapted for 7 days, mEC25 cells in the logarithmic growth phase were obtained, digested and centrifuged, and the cells were washed twice with PBS. Sufficient cells were resuspended in serum-free medium and matrix gel at a ratio of 1:1. mEC25 cells (cell suspension 100 μL, cell number about 6×10 6) were inoculated subcutaneously in the left axilla of 20 mice. After the growth of subcutaneous transplanted tumors stabilized, the mice were randomly divided into a tumor-bearing placebo group (Vehicle, n=5), a tumor-bearing low-dose drug group (Low dose, n=5), a tumor-bearing high-dose drug group (High dose, n=5) and a tumor-bearing cisplatin control group (CDDP, n=5). The drug administration method of each group was the same as the experimental scheme under "1.2.1.1 (1)".

[0136] (2) Detection methods and results:

[0137] The first day after inoculation of mEC25 cells was the first day. The size of the transplanted tumor was measured regularly, and the changes in the transplanted tumor volume over time were recorded. The body weight and diet were also recorded. Tumor volume: long diameter a, short diameter b, the formula for calculating tumor volume (TV) is: Tumor volume (TV) = 1 / 2 × a × b 2 About 2 weeks after administration, we killed the mice, collected samples, weighed them, and took photos.

[0138] The results are as follows Figure 4 As shown in Figures C and D, in vivo administration of Qiling Jiedu Fang can significantly inhibit the malignant growth of subcutaneous xenografts of mEC25 cells.

[0139] The above results indicate that in vivo administration of Qiling Jiedu Fang can significantly inhibit the malignant growth of subcutaneous transplanted ESCC cells.

[0140] 1.2.1.3 Safety test

[0141] (1) Experimental plan: To explore whether Qiling Jiedu Fang has obvious toxic and side effects in vivo. Six groups were set up: 2 groups without tumor transplantation, including healthy control group (HC, 200 μL of saline solution was given by gavage every day), toxic and side effect group (TS, 200 μL of Qiling Jiedu Fang granules in saline solution was given by gavage every day, the dose was 5.46 g / kg), and 4 groups with subcutaneous transplantation tumor models of KYSE150 cells were constructed, including Vehicle group (200 μL of saline solution was given by gavage every day), Low dose group (200 μL of Qiling Jiedu Fang granules in saline solution was given by gavage every day, the dose was 1.82 g / kg), High dose group (200 μL of Qiling Jiedu Fang granules in saline solution was given by gavage every day, the dose was 5.46 g / kg), and CDDP group (100 μL of cisplatin in saline solution was given by intraperitoneal injection every two days, the dose was 5 mg / kg).

[0142] (2) Detection methods and results:

[0143] ① From the first day of inoculation of KYSE150 cells, the weight and diet of mice were measured regularly. About 2 weeks after administration, the mice were killed, and samples were collected, weighed, and photographed.

[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 mice in the CDDP group was significantly reduced ( Figure 5 In addition, the organs of the mice were weighed, and the results showed that compared with the HC group, the kidney weight of the mice in the TS group was reduced, and there was no significant difference in the weight of the liver and spleen ( 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 HC group, TS group, Vehicle group and CDDP group was tested, and the blood routine indexes of white blood cell (WBC), platelet (PLT) and red blood cell (RBC) were compared. Blood routine test method: The mice were fasted for 12 hours before blood sample collection, and no water was allowed for the first hour. 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. Bowlinman (BM860) automatic analyzer was used for detection and analysis to detect the values ​​of white blood cell (WBC), platelet (PLT) and red blood cell (RBC) of mice.

[0146] The results are as follows Figure 5 As shown in FH, it can be seen that compared with the Vehicle group, the number of PLTs in the CDDP group was significantly reduced, and there was no significant effect on red blood cells and white blood cells. Compared with the HC group, oral administration of Qiling Jiedu Fang can significantly increase the platelets in the blood of mice, but has no significant effect on red blood cells and white blood cells.

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

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

[0149] (1) Experimental plan: To investigate the effect of Qiling Jiedu Recipe on the recurrence of esophageal cancer, a human cell line KYSE150 was selected to construct a mouse subcutaneous transplant tumor model. The subcutaneous transplant tumor model of KYSE150 cells was constructed as follows: After the nude mice were adapted for 7 days, KYSE150 cells in the logarithmic growth phase were obtained, digested and collected by centrifugation, and the cells were washed twice with PBS. Sufficient cells were resuspended in serum-free medium and KYSE150 cells (cell suspension 100 μL, cell number about 2×10 6 ) were inoculated subcutaneously in the left axilla of 28 nude mice. When the tumor volume grew to about 45 mm 3 When the tumor was shrunken to the size of a rice grain, cisplatin was given by intraperitoneal injection. When the tumor was retracted to the size of a rice grain, cisplatin administration was stopped. Then, when the tumor recurred and grew, the patients were randomly divided into 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 Fang granules in normal saline solution (dose of 1.82 g / kg) by gavage every day, the RH group was given 200 μL of Qiling Jiedu Fang granules in normal saline solution (dose of 5.46 g / kg) by gavage every day, and the RC group was given 100 μL of cisplatin in normal saline solution (dose of 5 mg / kg) by intraperitoneal injection every two days.

[0150] (2) Detection methods and results:

[0151] The volume of transplanted tumors was measured regularly, and the changes of transplanted tumors over time were recorded, and the body weight was also recorded. 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 24 days of administration, we killed the mice, took out the subcutaneous tumor, kidney, liver and other tissues of the mice, weighed them and took pictures.

[0152] ①Inhibitory effect on tumor recurrence

[0153] The results are as follows Figure 6 As shown in A, in vivo administration of Qiling Jiedu Fang can significantly inhibit the malignant growth of recurrent esophageal cancer tumors.

[0154] ②Safety results

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

[0156] In summary, oral administration of Qiling Jiedu Granules had no obvious toxic side effects, while after intraperitoneal injection of CDDP, the body weight and organs of mice underwent significant changes.

[0157] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A Chinese medicine composition for treating esophageal cancer of phlegm-qi type and recurrence, characterized in that: The invention comprises the following raw materials in parts by weight: 3-90 parts of astragalus, 1-60 parts of rubescens, 1-30 parts of codonopsis, 1-30 parts of raw atractylodes, 1-30 parts of tuckahoe, 1-30 parts of tangerine peel, 1-30 parts of bupleurum, 1-30 parts of clematis and 1-30 parts of roasted liquorice.

2. The Chinese medicine composition according to claim 1, characterized in that: The invention comprises 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 tuckahoe, 3-9 parts of tangerine peel, 3-9 parts of bupleurum, 3-9 parts of clematis and 3-9 parts of roasted liquorice.

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

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

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

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

7. The preparation method according to claim 3, 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.

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

9. Use of the Chinese medicine composition according to any one of claims 1 to 2 or the preparation method according to any one of claims 3 to 8 in preparing a drug for treating esophageal cancer or recurrence of esophageal cancer.

10. The use according to claim 9, characterized in that: The esophageal cancer is phlegm-qi intertwined type esophageal cancer.

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