Composition for regulating gastrointestinal function and treating gastrointestinal diseases and preparation method thereof

Abstract

The present invention discloses a composition for regulating gastrointestinal function and treating gastrointestinal diseases and a preparation method thereof. The composition comprises licorice, radish seeds, turmeric, beeswax, tuckahoe, scutellaria, konjac, cassia seed, chicken gizzard lining, kelp, aloe, hemp seed, broccoli, oligofructose, resistant dextrin, hericium erinaceus, sesame oil, lactic acid bacteria powder, stachyose, chitin, oligomaltose, Shenqu, inulin, probiotic fermentation powder, papain, and fucoidan. The composition can moisten the intestines and promote bowel movements, protect intestinal mucosa, regulate intestinal flora, inhibit Helicobacter pylori, remove intestinal garbage toxins, and decompose harmful carcinogens and excess fat. The composition has reasonable proportions of various components, has no obvious toxic and side effects and adverse reactions, and has high safety; it is convenient for patients to take and carry; the preparation process is simple and the production cost is low. Therefore, it has broad application prospects. Also disclosed is a polypeptide with a laxative effect and an amino acid sequence as shown in SEQ ID NO:1.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and specifically relates to a composition for regulating gastrointestinal function and treating gastrointestinal diseases, a polypeptide with a new structure, and their preparation methods and medical uses. Background Art

[0002] There are many types of gastrointestinal dysfunction or diseases, including constipation, gastrointestinal mucosal damage, Helicobacter pylori infection, etc.

[0003] Constipation refers to a type of symptom in which defecation is difficult due to hard or dry stool. Generally speaking, the frequency of normal bowel movements is about three times a day to three times a week. Complications of constipation include hemorrhoids, anal fissures, and fecal obstruction. Constipation usually has many causes, common causes include slow colon movement. The incidence of constipation is usually about 2-30%, and constipation is more common in the elderly than in the young. Epidemiological surveys have shown that people with constipation generally have problems with intestinal flora imbalance, including a decrease in the number of Lactobacillus and Bifidobacterium.

[0004] Intestinal moistening and laxative is a method used to treat constipation caused by aging and other reasons. In addition to drugs, specific intestinal moistening and laxative treatments also include dietary lifestyle adjustments and physical enema therapy as auxiliary treatments.

[0005] The intestinal mucosa is not only an important place for the digestion and absorption of nutrients, but also one of the portals for the invasion of pathogenic microorganisms and toxins. Under physiological conditions, the body can effectively prevent pathogenic microorganisms and toxins from crossing the intestinal mucosal barrier and invading tissues and organs outside the intestine through the intestinal mucosal epithelial barrier, immune barrier, microbial community barrier, chemical barrier, mucus layer and immobile water layer, so as to ensure that the body is in a healthy state. However, when the intestinal mucosal barrier is damaged, microorganisms and toxins in the intestine can penetrate the intestinal mucosal barrier and enter the portal vein and lymphatic system to cause bacterial translocation, and even develop into systemic inflammatory response syndrome and multiple organ failure. When the intestinal mucosa is damaged due to inflammation or trauma, it may cause intestinal stenosis, slow intestinal peristalsis, and cause constipation symptoms. Summary of the invention

[0006] In one aspect of the present invention, a composition for regulating gastrointestinal function and treating gastrointestinal diseases is provided, wherein the composition comprises the following components in parts by weight:

[0007] 20-95 weight parts of liquorice, 25-110 weight parts of radish seeds, 10-90 weight parts of turmeric, 25-100 weight parts of beeswax, 20-95 weight parts of tuckahoe, 10-90 weight parts of scutellaria, 25-100 weight parts of konjac, 20-95 weight parts of cassia seeds, 20-95 weight parts of chicken gizzard lining, 20-95 weight parts of kelp, 20-95 weight parts of aloe, 20-95 weight parts of hemp seeds, 20-95 weight parts of broccoli, 10-85 weight parts of oligofructose parts, resistant dextrin 10-90 parts by weight, hericium erinaceus 20-95 parts by weight, sesame oil 15-90 parts by weight, lactic acid bacteria powder 20-95 parts by weight, stachyose 25-95 parts by weight, chitin 20-90 parts by weight, isomaltooligosaccharide 10-95 parts by weight, Shenqu 10-90 parts by weight, inulin 25-100 parts by weight, probiotic fermentation powder 25-100 parts by weight, papain 20-90 parts by weight, and fucoidan 25-100 parts by weight.

[0008] In some specific embodiments of the present invention, the composition comprises the following components in parts by weight:

[0009] 70 parts by weight of liquorice, 80 parts by weight of radish seeds, 70 parts by weight of turmeric, 45 parts by weight of beeswax, 35 parts by weight of Poria cocos, 75 parts by weight of Scutellaria baicalensis, 40 parts by weight of konjac, 70 parts by weight of cassia seeds, 50 parts by weight of chicken gizzard lining, 60 parts by weight of kelp, 30 parts by weight of aloe, 40 parts by weight of hemp seeds, 40 parts by weight of broccoli, 40 parts by weight of oligofructose, 35 parts by weight of resistant dextrin, 75 parts by weight of Hericium erinaceus, 40 parts by weight of sesame oil, 40 parts by weight of lactic acid bacteria powder, 40 parts by weight of stachyose, 60 parts by weight of chitin, 80 parts by weight of isomaltooligosaccharide, 70 parts by weight of Shenqu, 70 parts by weight of inulin, 55 parts by weight of probiotic fermentation powder, 40 parts by weight of papain, and 60 parts by weight of fucoidan.

[0010] In some specific embodiments of the present invention, the probiotic fermentation powder is selected from: Lactobacillus bulgaricus, Lactobacillus reuteri, Lactobacillus plantarum, Lactobacillus rhamnosus, Bifidobacterium longum, Bifidobacterium bifidum, Lactobacillus acidophilus, Bifidobacterium lactis, Lactobacillus fermentum, Lactobacillus casei and Bifidobacterium infantis.

[0011] In some specific embodiments of the present invention, the probiotic fermentation powder is Lactobacillus bulgaricus or Lactobacillus reuteri. In some specific embodiments of the present invention, the probiotic fermentation powder is Lactobacillus bulgaricus. In some specific embodiments of the present invention, the probiotic fermentation powder is Lactobacillus reuteri.

[0012] In some specific embodiments of the present invention, the licorice is a dried root. In some specific embodiments of the present invention, the radish seed is a dried mature seed. In some specific embodiments of the present invention, the turmeric is a dried rhizome. In some specific embodiments of the present invention, the beeswax is Chinese honey bee wax. In some specific embodiments of the present invention, the Poria cocos is a dried sclerotium. In some specific embodiments of the present invention, the scutellaria is a dried root. In some specific embodiments of the present invention, the konjac is a dried underground tuber. In some specific embodiments of the present invention, the cassia seed is a dried mature seed of cassia. In some specific embodiments of the present invention, the kelp is a dried thallus of kelp. In some specific embodiments of the present invention, the aloe is a dried leaf. In some specific embodiments of the present invention, the hemp seed is a dried mature seed. In some specific embodiments of the present invention, the broccoli is a dried flower stem and flower bud. In some specific embodiments of the present invention, the oligofructose is purchased from Sigma-Aldrich. In some specific embodiments of the present invention, the resistant dextrin is purchased from Shaanxi Chenming Biological Co., Ltd. In some specific embodiments of the present invention, the Hericium erinaceus powder is purchased from Shaanxi Panier Biological Co., Ltd. In some specific embodiments of the present invention, the sesame oil is purchased from Shanghai Myrel Biochemical Technology Co., Ltd. In some specific embodiments of the present invention, the lactic acid bacteria powder is purchased from Jiangxi Ruiweier Biotechnology Co., Ltd. In some specific embodiments of the present invention, the stachyose is purchased from Shandong Fengtai Biological Co., Ltd. In some specific embodiments of the present invention, the chitosan is purchased from Shanghai Yuanye Biotechnology Co., Ltd. In some specific embodiments of the present invention, the isomaltooligosaccharide is purchased from Zhengzhou Yuhe Food Additive Co., Ltd. In some specific embodiments of the present invention, the Shenqu is Luyantang TCM. In some specific embodiments of the present invention, the inulin is the inulin of dahlia tubers. In some specific embodiments of the present invention, the bulgaricus Lactobacillus is purchased from Ningbo Mingzhou Biotechnology Co., Ltd. In some specific embodiments of the present invention, the papain is purchased from Shanghai Yuanye Biotechnology Co., Ltd. In some specific embodiments of the present invention, the fucoidan is purchased from MedChemExpress.

[0013] In some embodiments of the present invention, the composition comprises 0.4%-0.8% by weight of the polypeptide CW-P1, and the amino acid sequence of the polypeptide CW-P1 is shown in SEQ ID NO: 1.

[0014] In some embodiments of the present invention, the composition is composition CW-03 or CW-04 prepared by the method described in Example 1.

[0015] In some specific embodiments of the present invention, the preparation method of the composition is as described in Example 1.

[0016] In some specific embodiments of the present invention, the preparation method of the composition is as follows: 20-95 parts by weight of licorice, 25-110 parts by weight of radish seeds, 10-90 parts by weight of turmeric, 25-100 parts by weight of beeswax, 20-95 parts by weight of Poria cocos, 10-90 parts by weight of Scutellaria baicalensis, 25-100 parts by weight of konjac, 20-95 parts by weight of cassia seeds, 20-95 parts by weight of chicken gizzard lining, 20-95 parts by weight of kelp, 20-95 parts by weight of aloe vera, 20-95 parts by weight of hemp seeds and 20-95 parts by weight of broccoli are washed with water, air-dried at room temperature, and powdered. The mixture is crushed to an average particle size of 100 μm; and then uniformly mixed with 10-85 parts by weight of oligofructose, 10-90 parts by weight of resistant dextrin, 20-95 parts by weight of Hericium erinaceus, 15-90 parts by weight of sesame oil, 20-95 parts by weight of lactic acid bacteria powder, 25-95 parts by weight of stachyose, 20-90 parts by weight of chitin, 10-95 parts by weight of isomaltooligosaccharide, 10-90 parts by weight of Shenqu, 25-100 parts by weight of inulin, 25-100 parts by weight of probiotic fermentation powder, 20-90 parts by weight of papain and 25-100 parts by weight of fucoidan to obtain the product.

[0017] In some embodiments of the present invention, the composition is an oral formulation.

[0018] In some embodiments of the present invention, the dosage form of the composition is selected from tablets, capsules, granules, pills, oral liquids. In some embodiments of the present invention, the dosage form of the composition is tablets, capsules, granules or oral liquids. In some embodiments of the present invention, the composition is selected from oral tablets, capsules, granules, pills, oral liquids.

[0019] In some embodiments of the present invention, the method for preparing the composition comprises:

[0020] The composition of the present invention is irradiated and sterilized, granulated, dried and granulated to obtain granules.

[0021] In some embodiments of the present invention, the method for preparing the composition comprises:

[0022] The composition of the present invention is subjected to irradiation sterilization, granulation, drying and granulation to obtain granules; the granules are mixed with pharmaceutically acceptable excipients and then filled into a capsule shell or the granules are directly filled into a capsule shell to obtain a capsule.

[0023] In some embodiments of the present invention, the granulation method is to add 65%-75% ethanol in a volume fraction of 3-6% by weight to the sterilized mixture to make a soft material, and then granulate it through a 10-16 mesh sieve.

[0024] In some embodiments of the present invention, the drying temperature is 50° C.-70° C., and the drying time is 2-4 hours.

[0025] In some embodiments of the present invention, the sterilization is radiation sterilization, and the environment of the radiation sterilization is: cobalt-60 radiation sterilization, and the radiation dose is ≤6 kGy.

[0026] In some embodiments of the present invention, the composition is any one of the preparations described in Example 9.

[0027] In another aspect of the present invention, a preparation for regulating gastrointestinal function and treating gastrointestinal diseases is provided, characterized in that the preparation comprises the composition according to the present invention.

[0028] In some embodiments of the invention, the formulation is a pharmaceutical formulation.

[0029] In some embodiments of the invention, the formulation is an oral dosage form.

[0030] In some embodiments of the present invention, the preparation is in the form of tablets, capsules, granules, pills or oral liquids.

[0031] In some embodiments of the present invention, the preparation is selected from: granules, hard capsules, soft capsules, tablets.

[0032] In some embodiments of the present invention, the composition of the present invention is sterilized by irradiation, granulated, dried, and sized to obtain granules.

[0033] In some embodiments of the present invention, the composition of the present invention is sterilized by irradiation, granulated, dried, and sized to obtain granules; the granules are filled into capsule shells to obtain capsules.

[0034] In some embodiments of the present invention, the composition of the present invention is sterilized by irradiation, granulated, dried, and sized to obtain granules; the granules are loaded into soft capsule shells to obtain soft capsules.

[0035] In some embodiments of the present invention, the composition of the present invention is sterilized by irradiation, excipients are added, and tableting is performed to prepare tablets.

[0036] In some embodiments of the present invention, the preparation is any preparation as described in Example 9.

[0037] In another aspect of the present invention, a polypeptide is provided, characterized in that the amino acid sequence of the polypeptide is shown in SEQ ID NO: 1 or 2.

[0038] In some embodiments of the present invention, the amino acid sequence of the polypeptide is shown in SEQ ID NO: 1.

[0039] In another aspect of the present invention, provided is the use of the composition, the preparation and the polypeptide of the present invention in the preparation of a drug for regulating gastrointestinal function or treating gastrointestinal diseases.

[0040] In some embodiments of the present invention, use of the composition, preparation and polypeptide of the present invention in the preparation of a drug for regulating gastrointestinal function is provided.

[0041] In some embodiments of the present invention, there is provided use of the composition, the preparation and the polypeptide of the present invention in the preparation of a medicament for treating gastrointestinal diseases.

[0042] In some embodiments of the present invention, there is provided use of the composition, preparation and polypeptide of the present invention in the preparation of drugs for relieving constipation, protecting or repairing intestinal mucosa, regulating intestinal flora or resisting Helicobacter pylori.

[0043] In some embodiments of the present invention, the use of the composition, preparation and polypeptide of the present invention in the preparation of a laxative is provided. In some embodiments of the present invention, the use of the composition, preparation and polypeptide of the present invention in the preparation of a laxative is provided.

[0044] In some embodiments of the present invention, use of the composition, preparation and polypeptide of the present invention in the preparation of a drug for protecting intestinal mucosa is provided.

[0045] In some embodiments of the present invention, use of the composition, preparation and polypeptide of the present invention in the preparation of a medicament for regulating intestinal flora is provided. In some embodiments of the present invention, the regulating intestinal flora is to increase the content of intestinal bifidobacteria and lactobacilli.

[0046] In some embodiments of the present invention, use of the composition, preparation and polypeptide of the present invention in the preparation of anti-Helicobacter pylori drugs is provided.

[0047] In another aspect of the present invention, a method for preparing the composition of the present invention is provided, characterized in that the method for preparing the composition comprises: taking 20-95 parts by weight of licorice, 25-110 parts by weight of radish seeds, 10-90 parts by weight of turmeric, 25-100 parts by weight of beeswax, 20-95 parts by weight of tuckahoe, 10-90 parts by weight of scutellaria, 25-100 parts by weight of konjac, 20-95 parts by weight of cassia seeds, 20-95 parts by weight of chicken gizzard lining, 20-95 parts by weight of kelp, 20-95 parts by weight of aloe, 20-95 parts by weight of hemp seeds and 20-95 parts by weight of broccoli, washing them with water respectively, and then stirring at room temperature. The mixture is air-dried at room temperature and crushed to an average particle size of 100 μm; and then uniformly mixed with 10-85 parts by weight of oligofructose, 10-90 parts by weight of resistant dextrin, 20-95 parts by weight of Hericium erinaceus, 15-90 parts by weight of sesame oil, 20-95 parts by weight of lactic acid bacteria powder, 25-95 parts by weight of stachyose, 20-90 parts by weight of chitin, 10-95 parts by weight of isomaltooligosaccharide, 10-90 parts by weight of Shenqu, 25-100 parts by weight of inulin, 25-100 parts by weight of probiotic fermentation powder, 20-90 parts by weight of papain and 25-100 parts by weight of fucoidan to obtain the composition.

[0048] In some specific embodiments of the present invention, the preparation method of the composition is as described in Example 1.

[0049] In some embodiments of the present invention, the method for preparing the composition comprises:

[0050] The composition of the present invention is irradiated and sterilized, granulated, dried and granulated to obtain granules.

[0051] In some embodiments of the present invention, the method for preparing the composition comprises:

[0052] The composition of the present invention is subjected to irradiation sterilization, granulation, drying and granulation to obtain granules; the granules are mixed with pharmaceutically acceptable excipients and then filled into a capsule shell or the granules are directly filled into a capsule shell to obtain a capsule.

[0053] In some embodiments of the present invention, the granulation method is to add 65%-75% ethanol in a volume fraction of 3-6% by weight to the sterilized mixture to make a soft material, and then granulate it through a 10-16 mesh sieve.

[0054] In some embodiments of the present invention, the drying temperature is 50° C.-70° C., and the drying time is 2-4 hours.

[0055] In some embodiments of the present invention, the sterilization is radiation sterilization, and the environment of the radiation sterilization is: cobalt-60 radiation sterilization, and the radiation dose is ≤6 kGy.

[0056] The technical effects of the technical solution of the present invention are as follows:

[0057] Animal experiments show that the composition of the present invention can regulate gastrointestinal function. Specifically, the composition of the present invention can moisturize the intestines and promote bowel movements, protect and repair the intestinal mucosa, regulate intestinal flora, inhibit Helicobacter pylori, improve intestinal microecology, purify the gastrointestinal environment, remove intestinal waste toxins, decompose harmful carcinogens and excess fat, and improve the intestinal pH value and active intestinal peristalsis.

[0058] In addition, the composition of the present invention has reasonable proportions of various components, has no obvious toxic side effects and adverse reactions, is highly safe, is easy for patients to take and carry, has a simple preparation process, and has low production costs. Therefore, it has broad application prospects. DETAILED DESCRIPTION

[0059] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are selected according to conventional methods and conditions, or according to the product specifications. Unless otherwise specified, the reagents and raw materials used in the present invention are all commercially available.

[0060] Take 70g of licorice (dried root), 80g of radish seeds (dried mature seeds), 70g of turmeric (dried rhizomes), 45g of beeswax (Apis cerana), 35g of Poria (dried sclerotia), 75g of scutellaria (dried root), 40g of konjac (dried underground tuber), 70g of cassia seeds (dried mature seeds of cassia), 50g of chicken gizzard lining, 60g of kelp (dried fronds of kelp), 30g of aloe (dried leaves), 40g of hemp seeds (dried mature seeds), and 40g of broccoli (dried flower stems and flower buds), wash them with water, air-dry them at room temperature, and grind them to an average particle size of 100 μm. Mix them well to obtain a solid powder composition CW-01.

[0061] Take 40 grams of oligofructose (Sigma-Aldrich, F8052), 35 grams of resistant dextrin (Shaanxi Chenming Biological), 75 grams of Hericium erinaceus powder (Shaanxi Panier Biological), 40 grams of sesame oil (Shanghai Mairui Biochemical Technology), 40 grams of lactic acid bacteria powder (Jiangxi Ruiweier Biotechnology), 40 grams of stachyose (Shandong Fengtai Biological), 60 grams of chitin (Shanghai Yuanye Biotechnology), 80 grams of isomaltooligosaccharide (Zhengzhou Yuhe Food Additive Co., Ltd.), 70 grams of Shenqu (Luyantang TCM), 70 grams of inulin (inulin from dahlia tubers, Aladdin, product number I111088-250g), 55 grams of Lactobacillus bulgaricus (Ningbo Mingzhou Biotechnology Co., Ltd.), 40 grams of papain (Shanghai Yuanye Biotechnology), and 60 grams of fucoidan (MedChemExpress), mix well to obtain a solid powder composition CW-02.

[0062] Take 70g of licorice (dried root), 80g of radish seeds (dried mature seeds), 70g of turmeric (dried rhizomes), 45g of beeswax (Apis cerana), 35g of Poria (dried sclerotia), 75g of scutellaria (dried root), 40g of konjac (dried underground tuber), 70g of cassia seeds (dried mature seeds of cassia), 50g of chicken gizzard lining, 60g of kelp (dried fronds of kelp), 30g of aloe (dried leaves), 40g of hemp seeds (dried mature seeds), and 40g of broccoli (dried flower stems and flower buds), wash them separately with water, and air-dry them at room temperature. The mixture was dried and ground to an average particle size of 100 μm; then mixed with 40 g of oligofructose (Sigma-Aldrich, F8052), 35 g of resistant dextrin (Shaanxi Chenming Biological), 75 g of Hericium erinaceus powder (Shaanxi Panier Biological), 40 g of sesame oil (Shanghai Mairui Biochemical Technology), 40 g of lactic acid bacteria powder (Jiangxi Ruiweier Biotechnology), 40 g of stachyose (Shandong Fengtai Biological), 60 g of chitin (Shanghai Yuanye Biotechnology), 80 g of isomaltooligosaccharide (Zhengzhou Yuhe Food Additive Co., Ltd.), and 100 g of Shenqu (Luyantang Biotechnology). TCM) 70 g, inulin (inulin from dahlia tubers, Aladdin, item number I111088-250g) 70 g, Lactobacillus bulgaricus (Ningbo Mingzhou Biotechnology Co., Ltd.) 55 g, papain (Shanghai Yuanye Biotechnology) 40 g, and fucoidan (MedChemExpress) 60 g were mixed evenly to obtain a solid powder composition CW-03.

[0063] Take 70g of licorice (dried root), 80g of radish seeds (dried mature seeds), 70g of turmeric (dried rhizomes), 45g of beeswax (Apis cerana), 35g of Poria (dried sclerotia), 75g of scutellaria (dried root), 40g of konjac (dried underground tuber), 70g of cassia seeds (dried mature seeds of cassia), 50g of chicken gizzard lining, 60g of kelp (dried fronds of kelp), 30g of aloe (dried leaves), 40g of hemp seeds (dried mature seeds), and 40g of broccoli (dried flower stems and flower buds), wash them separately with water, and air-dry them at room temperature. The mixture was dried and ground to an average particle size of 100 μm; then mixed with 40 g of oligofructose (Sigma-Aldrich, F8052), 35 g of resistant dextrin (Shaanxi Chenming Biological), 75 g of Hericium erinaceus powder (Shaanxi Panier Biological), 40 g of sesame oil (Shanghai Mairui Biochemical Technology), 40 g of lactic acid bacteria powder (Jiangxi Ruiweier Biotechnology), 40 g of stachyose (Shandong Fengtai Biological), 60 g of chitin (Shanghai Yuanye Biotechnology), 80 g of isomaltooligosaccharide (Zhengzhou Yuhe Food Additive Co., Ltd.), and 100 g of Shenqu (Luyantang Biotechnology). The solid powder composition CW-04 was obtained by mixing 70 g of TCM, 70 g of inulin (inulin from dahlia tuber, Aladdin, product number I111088-250g), 55 g of Lactobacillus reuteri (LRC™, Lactobacillus reuteri NCIMB 30242), 40 g of papain (Shanghai Yuanye Biotechnology), and 60 g of fucoidan (MedChemExpress).

[0064] Adult female C57BL / 6 mice weighing 19.5-20.1g were randomly divided into groups, with 20 mice in each group. The modeling drug compound diphenoxylate was given by oral gavage to establish a constipation model in mice. The mice were gavaged with ink, and the time of the first black stool, the number of stools within 5 hours, and the stool weight were measured to reflect the defecation status of the model mice.

[0065] Group 1 was intragastrically administered with composition CW-01, Group 2 was intragastrically administered with composition CW-02, Group 3 was intragastrically administered with composition CW-03, and Group 4 was intragastrically administered with composition CW-04; mice in Groups 1 to 4 were intragastrically administered once a day.

[0066] Every morning, groups 1 to 4 were gavaged with a uniform suspension of each composition in sterile water (drug concentration 2 mg / mL, prepared by stirring each drug in sterile water at 300 r / min at room temperature for 10 min), with a dosing volume of 10 mL / kg body weight. The normal control group and model group mice were given 10 mL / kg body weight of normal saline.

[0067] After continuous gavage for 10 days, mice in each group were fasted but not watered for 20 hours. The normal control group was given saline, and groups 1-4 and the model group were gavaged with 25 mg diphenoxylate / kg body weight of compound diphenoxylate tablets (Changzhou Kangpu Pharmaceutical) suspended in water (difoxylate concentration is about 2.5 mg / mL). After 30 minutes, mice in each group were gavaged with an equal amount of ink. From the beginning of the gavage, the time of discharge of the first black stool of each mouse, the number and weight of black stools in different time periods were recorded. As shown in Table 1, the data are the average values ​​of mice in the group. The weight of black stool within 12 hours in group 3 (gavage administration of composition CW-03) and group 4 (gavage administration of composition CW-04) was significantly higher than that in the model group, group 1 and group 2, respectively, with statistically significant differences (P<0.01); in addition, the discharge time of the first black stool in group 3 and group 4 was significantly shorter than that in the model group, group 1 and group 2 (P<0.01), and the number of black stool particles within 12 hours was significantly more than that in the model group, group 1 and group 2 (P<0.01). This shows that composition CW-03 and composition CW-04 have strong laxative function. The combination of CW-01 and CW-02 has a synergistic effect and has achieved unexpected technical effects.

[0068] Table 1. Laxative function test results

[0069] Time to discharge of the first black stool (min) Number of black stools within 12 hours (grains) Weight of black stool within 12 hours (g) Normal control group 75.9 65.6 0.65 Model Group 163.4 26.7 0.17 Group 1 (Composition CW-01) 132.1 33.0 0.29 Group 2 (Composition CW-02) 127.6 35.3 0.32 Group 3 (Composition CW-03) 76.3 67.4 0.71 Group 4 (Composition CW-04) 67.5 63.0 0.78

[0070] Eight-week-old healthy clean-grade male BALB / c mice weighing 20.5-21.1 g were selected. They were fed with a standard diet for 1 week before the experiment and randomly divided into sham injury group, simple burn group, and burn + CW-03 group, with 10 mice in each group.

[0071] After disinfection of the skin of the left lower abdomen of each group of mice, sodium pentobarbital was intraperitoneally injected for anesthesia. The backs were shaved. The backs of mice in the simple burn group and the burn + CW-03 group were placed in 90℃ hot water for 12 seconds, resulting in 30% TBSA Ⅲ-degree scald on the back (hereinafter referred to as burn); the backs of mice in the sham injury group were placed in 37℃ warm water for 12 seconds to cause sham injury. Lactated Ringer's solution was immediately injected intraperitoneally in each group of mice for resuscitation. After the mice woke up, the mice in the burn + CW-03 group were gavaged with 2mg / mL CW-03 suspension (10mL / kg body weight), and the other two groups of mice were gavaged with 10mL / kg body weight of normal saline, twice a day, for 10 days. The small intestinal tissues of mice in each group at different time points after injury were obtained, fixed with polyformaldehyde, embedded in paraffin, sectioned, and HE stained with fluorescence microscopy to observe the pathological changes of small intestinal tissue.

[0072] The mice in the sham injury group had no congestion or edema in the intestinal mucosa at any time point after injury. The mice in the simple burn group had severe intestinal mucosal damage 10 days after injury, and all mice in this group had congestion and edema in the intestinal mucosa. No intestinal mucosal edema and congestion were observed in all mice in the burn + CW-03 group 10 days after injury. This indicates that the composition CW-03 can accelerate the repair of intestinal mucosal damage.

[0073] Male Kunming mice (weight 19.1-20.4 g) were randomly divided into two groups, including a normal control group and a CW-03 drug group, with 10 mice in each group.

[0074] 0.1g of feces from each mouse was aseptically collected to check the intestinal flora (Bifidobacterium, Lactobacillus), and the appropriate dilution was selected and inoculated on the culture medium. After culture, the colonies were identified and counted by colony morphology, Gram staining microscopy, biochemical reaction, etc., and the number of bacteria per gram of wet feces was calculated as the colony data before the test sample was administered. The composition CW-03 was administered by gavage once a day for 14 consecutive days. 24h after the last administration of the test sample, 0.1g of feces from each mouse was aseptically collected again, and the intestinal flora was detected by the same method.

[0075] The results showed that the levels of bifidobacteria and lactobacilli in the intestines of the normal control group mice did not increase before and after administration; while the levels of bifidobacteria and lactobacilli in the intestines of the CW-03 group mice increased by 9.7% and 3.3% respectively after administration compared with those before administration (with statistically significant differences, P<0.05), and also increased significantly compared with the normal control group mice after administration (P<0.05).

[0076] Normal, healthy 7-week-old BALB / c mice were randomly divided into normal control group, pathological model group and drug administration group, with 20 mice in each group.

[0077] The pathological model group and the drug-treated group were intragastrically administered with 0.15 mL of 0.2 mol / L sodium bicarbonate solution, and 15 minutes later, the concentration of 1×10 9 CFU / mL Helicobacter pylori 0.1mL; this operation was repeated once a day. After continuous gavage for 8 days, the model was successfully established. The normal control group was replaced with an equal volume of normal saline instead of Helicobacter pylori solution.

[0078] Then, the dosing group was gavaged with a uniform suspension of the composition CW-03, CW-01 or CW-02 in sterile water (drug concentration 10 mg / mL) every morning, and the dosing volume was 10 mL / kg body weight. The mice in the normal control group and the pathological model group were given 10 mL / kg body weight of normal saline. After 20 consecutive days of gavage, the mice were killed, and the Helicobacter pylori infection after treatment was tested using a rapid urease test. The effective rate (%) was calculated by dividing the number of urease-negative animals after treatment by the total number of animals in each group. The experiment was repeated 3 times, and the results were averaged. As shown in Table 2, the effective rate of the CW-03 administration group was significantly higher than that of the model group, CW-01 and CW-02 groups, with a statistically significant difference (P<0.05). It was unexpected that the combination of CW-01 and CW-02 also had a synergistic effect in anti-Helicobacter pylori.

[0079] Table 2. Anti-Helicobacter pylori test results

[0080] Effectiveness (%) Model Group 0% Dosage group (composition CW-01) 15.0% Administration group (composition CW-02) 13.3% Administration group (composition CW-03) 65.0%

[0081] The solid powder composition CW-03 prepared by the method described in Example 1 was separated and purified using an InfinityLabPursuit XRs C18 Preparative HPLC Column (acetonitrile-water gradient elution) to obtain a polypeptide CW-P1 (SEQ ID NO: 1) with a mass percentage of 0.6% and a polypeptide CW-P2 (SEQ ID NO: 2) with a mass percentage of 0.4%. The structural characterization data of the polypeptide CW-P1 (SEQ ID NO: 1) and the polypeptide CW-P2 (SEQ ID NO: 2) are the same as those in the following examples.

[0082] The crude products of peptides CW-P1 and CW-P2 were obtained by Fmoc solid phase synthesis using a peptide synthesizer (Aurora, VERSA 110).

[0083] The crude product was dried at 20°C under vacuum conditions; separated and purified using an InfinityLab Pursuit XRs C18Preparative HPLC Column (acetonitrile-water gradient elution) and freeze-dried to obtain polypeptides CW-P1 (SEQ ID NO: 1) and CW-P2 (SEQ ID NO: 2) with HPLC purities of 95.6% and 95.1%, respectively.

[0084] Elemental analysis showed that the test values ​​of peptides CW-P1 and CW-P2 were consistent with the theoretical values. ESI-MS showed that peptide CW-P1 had a main ion peak of [M+H]+ 2159.0, and mass spectrometry analysis showed that its ion peak was consistent with

[0085] Molecular structure. ESI-MS showed that the peptide CW-P2 had a main ion peak of [M+H]+ 2111.0, and mass spectrometry analysis showed that its ion peak was consistent with the molecular structure.

[0086] Polypeptide CW-P1: TMHNETGTQRKTGDEARNI (SEQ ID NO: 1).

[0087] Peptide CW-P2: QFTGQSHADEVRISNHAIT (SEQ ID NO: 2)

[0088] The same method as in Example 2 was used for testing, except that the polypeptide was used instead of each composition for oral administration. The results are shown in Table 3, which shows that the polypeptide CW-P1 significantly improved the time of first black stool excretion, the number of black stool particles and the weight of the mice compared with the model group, with statistically significant differences (P<0.01).

[0089] Table 3. Laxative function test results

[0090] Time to discharge of the first black stool (min) Number of black stools within 12 hours (grains) Weight of black stool within 12 hours (g) Normal control group 76.3 68.1 0.69 Model Group 177.9 23.0 0.18 Peptide CW-P1 group 102.1 53.0 0.59 Peptide CW-P2 group 161.5 25.2 0.21

[0091] Preparation Example 1: The composition CW-03 prepared as described in Example 1 is sterilized by irradiation, granulated, dried, and sized to obtain granules.

[0092] Preparation Example 2: The composition CW-03 prepared as described in Example 1 is subjected to irradiation sterilization, granulation, drying, and granulation to obtain composition granules; the composition granules are loaded into soft capsule shells, with each capsule shell being loaded with 300 mg of medicine, to obtain soft capsules.

[0093] Preparation Example 3: The composition CW-03 prepared as described in Example 1 was irradiated and sterilized with 60Co radiation at a dose of 4 kGy; 5% by weight of 70% ethanol (V / V) was added to the sterilized mixture to make a soft material, which was then granulated through a 14-mesh sieve; dried at 60 degrees Celsius for 3 hours; and granulated to obtain granules.

[0094] Preparation Example 4: The composition CW-03 prepared by the method described in Example 1 is irradiated and sterilized with 60Co radiation and an irradiation dose of 4 kGy; 5% by weight of 70% ethanol (V / V) is added to the sterilized mixture to make a soft material and then granulated through a 14-mesh sieve; dried at 60 degrees Celsius for 3 hours; granulated to obtain composition granules; the composition granules are loaded into capsule shells, each capsule shell is loaded with 300 mg of medicine, to obtain soft capsules.

[0095] Although the embodiments of the present disclosure have been described above, it will be understood that the above embodiments are exemplary and do not constitute limitations to the present disclosure. Any modifications, equivalent substitutions and improvements made by ordinary technicians in the field within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A composition for regulating gastrointestinal function and treating gastrointestinal diseases, characterized in that: The composition is composed of the following components in parts by weight: 20-95 weight parts of liquorice, 25-110 weight parts of radish seeds, 10-90 weight parts of turmeric, 25-100 weight parts of beeswax, 20-95 weight parts of tuckahoe, 10-90 weight parts of scutellaria, 25-100 weight parts of konjac, 20-95 weight parts of cassia seeds, 20-95 weight parts of chicken gizzard lining, 20-95 weight parts of kelp, 20-95 weight parts of aloe, 20-95 weight parts of hemp seeds, 20-95 weight parts of broccoli, 10-85 weight parts of oligofructose parts, resistant dextrin 10-90 parts by weight, Hericium erinaceus 20-95 parts by weight, sesame oil 15-90 parts by weight, lactic acid bacteria powder 20-95 parts by weight, stachyose 25-95 parts by weight, chitin 20-90 parts by weight, isomaltooligosaccharide 10-95 parts by weight, Shenqu 10-90 parts by weight, inulin 25-100 parts by weight, probiotic fermentation powder 25-100 parts by weight, papain 20-90 parts by weight, and fucoidan 25-100 parts by weight; The probiotic fermentation powder is Lactobacillus bulgaricus.

2. The composition according to claim 1, characterized in that The composition is composed of the following components in parts by weight: 70 parts by weight of liquorice, 80 parts by weight of radish seeds, 70 parts by weight of turmeric, 45 parts by weight of beeswax, 35 parts by weight of Poria cocos, 75 parts by weight of scutellaria, 40 parts by weight of konjac, 70 parts by weight of cassia seeds, 50 parts by weight of chicken gizzard lining, 60 parts by weight of kelp, 30 parts by weight of aloe, 40 parts by weight of hemp seeds, 40 parts by weight of broccoli, 40 parts by weight of oligofructose, 35 parts by weight of resistant dextrin, 75 parts by weight of Hericium erinaceus, 40 parts by weight of sesame oil, 40 parts by weight of lactic acid bacteria powder, 40 parts by weight of stachyose, 60 parts by weight of chitin, 80 parts by weight of isomaltooligosaccharide, 70 parts by weight of Shenqu, 70 parts by weight of inulin, 55 parts by weight of probiotic fermentation powder, 40 parts by weight of papain, and 60 parts by weight of fucoidan; The probiotic fermentation powder is Lactobacillus bulgaricus.

3. A preparation for regulating gastrointestinal function and treating gastrointestinal diseases, characterized in that: The preparation comprises the composition as claimed in claim 2; The preparation is an oral dosage form, which is a tablet, capsule, granule, pill or oral liquid.

4. Use of the composition according to claim 1 or 2 or the preparation according to claim 3 in the preparation of medicaments for relieving constipation, repairing intestinal mucosa, regulating intestinal flora or resisting Helicobacter pylori.

5. The method for preparing the composition according to claim 1, characterized in that: The preparation method of the composition is: Take 20-95 parts by weight of liquorice, 25-110 parts by weight of radish seeds, 10-90 parts by weight of turmeric, 25-100 parts by weight of beeswax, 20-95 parts by weight of tuckahoe, 10-90 parts by weight of scutellaria, 25-100 parts by weight of konjac, 20-95 parts by weight of cassia seeds, 20-95 parts by weight of chicken gizzard lining, 20-95 parts by weight of kelp, 20-95 parts by weight of aloe, 20-95 parts by weight of hemp seeds and 20-95 parts by weight of broccoli, wash them with water respectively, air-dry them at room temperature, and grind them into average particle size of 100 microns; then The composition is obtained by uniformly mixing 10-85 parts by weight of oligofructose, 10-90 parts by weight of resistant dextrin, 20-95 parts by weight of Hericium erinaceus, 15-90 parts by weight of sesame oil, 20-95 parts by weight of lactic acid bacteria powder, 25-95 parts by weight of stachyose, 20-90 parts by weight of chitin, 10-95 parts by weight of isomaltooligosaccharide, 10-90 parts by weight of Shenqu, 25-100 parts by weight of inulin, 25-100 parts by weight of probiotic fermentation powder, 20-90 parts by weight of papain and 25-100 parts by weight of fucoidan.

6. The preparation method according to claim 5, further comprising the following steps: The composition is irradiated and sterilized, granulated, dried, and sized to obtain granules; or The composition is sterilized by irradiation, granulated, dried, and granulated to obtain granules, and the granules are mixed with pharmaceutically acceptable excipients and then filled into capsule shells or the granules are directly filled into capsule shells to obtain capsules.

7. The preparation method according to claim 6, wherein the granulation method is to add 65%-75% ethanol in a volume fraction of 3-6% by weight to the sterilized mixture to make a soft material, and then granulate it through a 10-16 mesh sieve; The drying temperature is 50°C-70°C and the drying time is 2-4 hours; The sterilization is radiation sterilization, and the environment of the radiation sterilization is: cobalt-60 radiation sterilization, and the radiation dose is ≤6 kGy.