A medicine for irritable bowel syndrome and preparation method thereof

Abstract

The invention discloses a medicine for irritable bowel syndrome and a preparation method thereof. The medicine is prepared from Radix Pseudostellariae, Pericarpium Citri Reticulatae, Rhizoma Acorus Calamus, Radix Glechomae and Oxalis Sorrel, and the preparation method is prepared by water extraction. The medicine can significantly improve the propulsion rate of the small intestine, promote intestinal peristalsis, and is used to treat irritable bowel syndrome such as diarrhea, gastrointestinal spasm, and constipation. The medicine has the advantages of slow drug release, prolonged drug efficacy, reduced toxicity, reduced adverse reactions, etc., and the preparation method is simple, and it is one of the more ideal Chinese medicine dosage forms, which not only retains the characteristics of the original Chinese medicine formula, but also is convenient for patients to carry and take.

Description

Field of the Invention

[0001] The invention relates to a medicine for irritable bowel syndrome and a preparation method thereof, and belongs to the field of pharmaceutical technology. Background Art

[0002] Irritable bowel syndrome is one of the more common diseases. Currently, Western medicine mostly uses surgery for treatment, which is expensive and has certain side effects. For patients who are not suitable for surgery or do not accept surgery, the application of traditional Chinese medicine treatment is the best choice. It belongs to the category of "diarrhea", "constipation" and "abdominal pain" in traditional Chinese medicine. Its causes are mainly congenital deficiency and (or) acquired malnutrition, emotional disorders leading to liver wood riding spleen, improper diet, excessive work and rest, and exposure to external evil. The pathogenesis is spleen and stomach weakness and (or) liver failure to dredge. Traditional Chinese medicine prescriptions have good efficacy in the treatment of irritable bowel syndrome, with few side effects, safety and reliability, and suitable for long-term use. However, most Chinese medicine prescriptions are decoctions, which have the disadvantages of easy mold and deterioration after long-term storage, large volume for direct use, and inconvenient to carry.

[0003] The Radix Pseudostellariae, Pericarpium Citri Reticulatae and Acorus Tatarinowii medicinal materials of the present invention are varieties included in the first volume of the Chinese Pharmacopoeia, and the Radix Glechomae and Oxalis Herba medicinal materials are varieties included in the Quality Standards for Chinese Medicinal Materials and Ethnic Medicinal Materials of Guizhou Province (2003 Edition); all of them have no toxicity, incompatibility taboos and dietary taboos. The present invention studies the extraction process of the effective components of the prescription and prepares them into pills, providing theoretical support for the development of the treatment of irritable bowel syndrome and providing scientific and reliable process parameters for industrial production. The curative effect is remarkable and the quality of life is improved. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide a medicine for irritable bowel syndrome and a preparation method thereof. The present invention is used to treat irritable bowel syndrome such as diarrhea, gastrointestinal spasm, constipation, etc. The preparation method of the present invention is easy to operate, can ensure that the effective ingredients of the product are not lost, and has significant therapeutic effect.

[0005] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0006] A medicine for irritable bowel syndrome, wherein the medicinal effective ingredients are calculated by weight: 20-100 parts of Pseudostellaria heterophylla, 20-100 parts of dried orange peel, 20-100 parts of Acorus calamus, 50-130 parts of Cynanchum cyrtonema and 20-100 parts of Oxalis sorrel.

[0007] The above-mentioned medicine for irritable bowel syndrome, its medicinal active ingredients are calculated by weight: 40-80 parts of Pseudostellaria heterophylla, 40-80 parts of dried tangerine peel, 40-80 parts of Acorus calamus, 70-110 parts of Glechoma longituba, and 40-80 parts of Oxalis sorrel.

[0008] Specifically, the aforementioned medicine for irritable bowel syndrome, its medicinal active ingredients are calculated by weight: 60 parts of Pseudostellaria heterophylla, 60 parts of dried tangerine peel, 60 parts of Acorus calamus, 90 parts of Glechoma longituba, and 60 parts of Oxalis sorrel.

[0009] The preparation method of the aforementioned drug for irritable bowel syndrome is carried out according to the following steps:

[0010] (1) Weigh all the medicinal materials in the prescription according to the proportion, add 6-12 times the amount of aqueous solution, reflux extraction 1-4 times, each time for 0.5-2 hours, filter, combine the filtrate, concentrate the filtrate to a relative density of 1.20-1.26, vacuum dry at 60-100° C. to form a dry extract, grind into fine powder, add 0.9-1.3 times the amount of soluble starch, mix, and obtain product A;

[0011] (2) Weigh 2-5% of the total weight of product A and add low-substituted hydroxypropyl cellulose to product A and mix well to obtain product B;

[0012] (3) Weigh 10-20% of the total weight of product B as vinegar as a wetting agent, add it to product B and mix well, make pills by plasticizing, pass through a pill making machine, make 1000g pills, and vacuum dry at 50-80°C for 12-16h to obtain the finished product.

[0013] Specifically, the above preparation method is carried out according to the following steps:

[0014] (1) Weigh all the medicinal materials in the prescription according to the proportion, add 8-12 times the amount of aqueous solution, reflux extraction 2-4 times, each time for 0.5-1.5 hours, filter, combine the filtrate, concentrate the filtrate to a relative density of 1.1-1.3, vacuum dry at 70-90° C. to form a dry extract, grind into fine powder, add 1.0-1.5 times the amount of soluble starch, and obtain product A;

[0015] (2) Weigh 2-5% of the total weight of product A and add low-substituted hydroxypropyl cellulose to product A and mix well to obtain product B;

[0016] (3) Weigh 10-20% of the total weight of product B as vinegar as a wetting agent, add it to product B and mix well, make pills by plasticizing, pass through a pill making machine, make 1000g pills, and vacuum dry at 60-70°C for 13-15h to obtain the finished product.

[0017] Specifically, the above preparation method is carried out according to the following steps:

[0018] (1) Weigh all the medicinal materials in the prescription according to the proportion, add 10 times the amount of aqueous solution, reflux extraction 3 times, each time for 1 hour, filter, combine the filtrate, concentrate the filtrate over direct fire to a relative density of 1.22-1.24, dry it in vacuum at 80°C to form a dry extract, grind it into fine powder, add 1.1 times the amount of soluble starch, and obtain product A;

[0019] (2) Weigh 2.2% of the total weight of product A and add it to product A and mix well to obtain product B;

[0020] (3) Weigh 17% of the total weight of product B as vinegar as a wetting agent, add it to product B and mix well, make pills by plasticizing, pass through a pill making machine, make 1000g pills, and vacuum dry at 65℃ for 14h to obtain the finished product.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] 1. The drug for irritable bowel syndrome of the present invention is made of Radix Pseudostellariae, dried orange peel, Acorus calamus, Radix Glechomae, and Oxalis. Radix Pseudostellariae is the dried root of Psudostellaria heterophylla (Miq.) Pax ex Pax et Ho ffm., a plant of Caryophyllaceae; it is sweet, slightly bitter, and flat. It enters the spleen and lung meridians; it has the effects of invigorating qi and strengthening spleen, promoting body fluid and moistening lungs; it mainly treats symptoms such as spleen deficiency, fatigue, loss of appetite, weakness after illness, deficiency of qi and yin, spontaneous sweating and thirst, dry cough due to lung dryness, etc.; it mainly contains chemical components such as cyclic peptides, glycosides, sugars, amino acids, phospholipids, volatile oils, fatty acids, oils, sterols, and trace elements; and it has pharmacological effects such as myocardial protection, immunomodulation, anti-oxidation, protection of intestines and kidneys, anti-stress, and lipid-lowering. Dried orange peel is the dried mature peel of Citrus reticulata Blanco, a plant of Rutaceae, and its cultivated varieties. Bitter, pungent, warm; enters the lung and spleen meridians. Qi-strengthening, phlegm-drying; used for abdominal distension, poor appetite, vomiting and diarrhea, cough and phlegm; Tangerine peel is mainly composed of flavonoids, and also contains limonoids, alkaloids and volatile oils; it has pharmacological effects such as inhibiting the gastrointestinal tract, anti-tumor, anti-oxidation, preventing diabetes, anti-allergy, antitussive, anti-asthmatic, and anti-allergic inflammation. Acorus tatarinowii Schott is the dried rhizome of Acorus tatarinowii Schott, a plant of the Araceae family; pungent, bitter, warm, enters the heart and stomach meridians; opens the mind and eliminates phlegm, wakes up the mind and improves intelligence, removes dampness and stimulates appetite; forgetfulness and insomnia, tinnitus and deafness, abdominal distension without hunger, and dysentery; the main chemical components are volatile oils, organic acids, flavonoids, terpenes, etc., and have pharmacological effects such as sedation, anti-oxidation, lowering blood lipids, anti-thrombosis, anti-tumor, and antibacterial. Radix Polygoni Multiflori is the dried root of Cynanchum auriculatum Royle ex Wight, a plant of the Apocynaceae family. It is sweet, slightly bitter, and neutral in nature. It enters the liver, kidney, spleen, and stomach meridians. It is also known as Taishan Polygoni Multiflori, Polygoni Multiflori (Shandong), ground gourd, mountain gourd, wild yam, and white yam. It is mainly produced in Guizhou and is used by ethnic minorities in the province. It has the effects of nourishing yin, tonifying deficiency, strengthening the spleen and promoting digestion, and is mainly used for exhaustion, dysentery, malnutrition, fullness and stomach pain, leucorrhea, and sores and ringworms. Its main chemical components include steroidal saponins, terpenes, polysaccharides, alkaloids, and other components. It has pharmacological effects such as anti-tumor, bidirectional immunomodulation, antioxidant, liver protection, anti-inflammatory, antibacterial, and promoting intestinal peristalsis.Oxalis corniculata L. is the fresh or dried whole herb of the Oxalis corniculata L. of the Oxalidaceae family. It is sour and cold and enters the liver, lung and bladder meridians. It is also used by ethnic minorities in our province. It has the effects of clearing away heat and dampness, cooling blood and reducing swelling, detoxifying and dispersing blood stasis. It is used for diarrhea caused by damp heat, stranguria, leucorrhea, sore throat, furuncle, carbuncle, eczema, measles, falls, burns, and insect bites. Modern pharmacology and clinical studies have shown that Oxalis corniculata not only has the effects of anti-inflammatory and bactericidal, promoting blood circulation and removing blood stasis, but also has anti-tumor, antioxidant, and liver-protecting activities. Oxalis contains many types of chemical components, including flavonoids, phenolic acids, phenylpropanoids, and alkaloids. It has the effects of analgesia and anti-inflammatory, protecting the liver, anti-hyperlipidemia, antioxidant, affecting the digestive system, anti-tumor, and antibacterial.

[0023] Explanation of the prescription: The prescription uses Geshanxiao, which is sweet and slightly bitter, and is in the liver, kidney, spleen, and stomach meridians. It nourishes yin and replenishes deficiency, strengthens the spleen and digests food, and is the main medicine. The minister uses Taizishen, which is sweet and flat, and is in the spleen and stomach meridians, which can replenish qi and strengthen the spleen, promote fluid and moisten the lungs, and help the main medicine nourish spleen yin, benefit spleen qi, and eliminate food stagnation; the auxiliary uses Shichangpu, which is pungent, fragrant, warm and unblocked, relieves depression, eliminates phlegm, wakes up the mind, and eliminates dampness, stimulates appetite and improves intelligence; Chenpi regulates qi, dries dampness and harmonizes the stomach, and its aromatic and damp-removing properties strengthen the warming and drying and damp-removing power of calamus, and the two are used together to promote the spleen's transportation and transformation power; and then uses Oxalis, which is sour and cold, clears heat and promotes dampness, cools blood and reduces swelling, and detoxifies and disperses blood stasis. Its sour and soft properties are in the liver meridian, and together with the two main and minister drugs, it can achieve the effects of relieving pain, supporting the spleen and suppressing the liver, and is a total adjuvant. The whole prescription is nourishing and flowing, and the sour and sweet are combined. The combination of various drugs plays the role of supporting the spleen and softening the liver, strengthening the spleen and stopping diarrhea.

[0024] 2. Studies have found that the water extract of Radix Pseudostellariae has significant effects on protecting the intestines, kidneys, myocardial protection, immunomodulation, antioxidant and anti-stress. The water extract of dried orange peel has a significant effect on the digestive system and inhibits gastrointestinal contraction and spasm. The water extract of Acorus tatarinowii has obvious sedative, antioxidant, antibacterial, and digestive system effects. The water extract of Herba Lycopodii has significant effects on promoting gastric emptying, intestinal peristalsis, and treating indigestion. The water extract of Herba Vinegare has obvious anti-inflammatory and analgesic effects, protects the liver, and is antioxidant, and affects the digestive system. In summary, the present invention selects water as the extraction solvent.

[0025] 3. Among the drugs for irritable bowel syndrome proposed by the present invention, the content of β-EP, PGE2, cAMP, serum 5-hydroxytryptamine (5-HT), and SP in the serum of mice subjected to the acetic acid writhing experiment was detected by ELISA method. It was found that the medium and high dose groups of the pills of the present invention had the most significant effect on the down-regulation of β-EP, PGE2, cAMP, serum 5-hydroxytryptamine (5-HT), and SP levels in the serum of mice, and exerted anti-inflammatory and analgesic effects. The content of 5-HT in serum and cerebral cortex changes, and it is an active substance for analgesia in the central nervous system. The present invention can significantly improve the propulsion rate of the small intestine and promote intestinal peristalsis.

[0026] 4. The pellets of the drug obtained by the preparation process of the present invention are round and smooth, with uniform color and size, and the moisture content and dissolution time limit meet the requirements, and RSD is less than 5%, and the hesperidin content can reach 6 mg / g, the transfer rate exceeds 20%, and the average transfer rate reaches 21.135%. The prescription process is stable, the quality is controllable, and it is reasonable and feasible.

[0027] 5. The present invention prepares the formulated drugs into pills, which have the advantages of slow drug release, prolonged drug efficacy, reduced toxicity, reduced adverse reactions, etc., and the preparation method is simple. It is one of the more ideal Chinese medicine dosage forms, which not only retains the characteristics of the original Chinese medicine formula, but also is convenient for patients to carry and take. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 :Experimental design flow chart;

[0029] Figure 2 : Water absorption investigation chart. DETAILED DESCRIPTION

[0030] The present invention will be further described below in conjunction with the embodiments, but they are not intended to limit the present invention.

[0031] Example 1.

[0032] Prescription: 6g of Pseudostellaria heterophylla, 6g of dried tangerine peel, 6g of Acorus calamus, 9g of Gynostemma pentaphyllum, and 6g of Oxalis sorrel.

[0033] Process: Weigh all the medicinal materials in the prescription according to the proportion, add 10 times the amount of aqueous solution, reflux extraction 3 times, 1 hour each time, filter, combine the filtrate, concentrate the filtrate to a relative density of 1.23, vacuum dry at 80°C to form a dry extract, grind into fine powder, add 1.1 times the soluble starch, then add 2.2% of the total weight of low-substituted hydroxypropyl cellulose, mix well, then add 17% of the total weight of vinegar as a wetting agent, mix well, make pills by plastic method, pass through a pill making machine, make 1000g pills, and vacuum dry at 65°C for 14 hours.

[0034] Dosage and Administration: Take the medicine three times a day, 20 pills each time.

[0035] Example 2.

[0036] Prescription: 2g of Pseudostellaria heterophylla, 2g of dried tangerine peel, 2g of Acorus calamus, 5g of Gynostemma pentaphyllum, and 2g of Oxalis sorrel.

[0037] Process: Weigh all the medicinal materials in the prescription according to the proportion, add 6 times the amount of aqueous solution, reflux extraction once, each time for 0.5 h, filter, combine the filtrate, concentrate the filtrate to a relative density of 1.20, vacuum dry at 60°C to form a dry extract, grind into fine powder, add 0.9 times the soluble starch, mix, add 2% of the total weight of low-substituted hydroxypropyl cellulose, mix well, add 15% of the total weight of vinegar as a wetting agent, mix well, make pills by plastic method, pass through a pill making machine, make 1000g pills, vacuum dry at 50°C for 12 h to obtain the finished product.

[0038] Dosage and Administration: Take the medicine three times a day, 20 pills each time.

[0039] Example 3.

[0040] Prescription: Made from 10g of Pseudostellaria heterophylla, 10g of dried tangerine peel, 10g of Acorus calamus, 13g of Gynostemma pentaphyllum, and 10g of Oxalis sorrel.

[0041] Process: Weigh all the medicinal materials in the prescription according to the proportion, add 12 times the amount of aqueous solution, reflux extraction 4 times, 2 hours each time, filter, combine the filtrate, concentrate the filtrate to a relative density of 1.26, vacuum dry at 100°C to form a dry extract, grind into fine powder, add 1.3 times the soluble starch, mix and add 5% of the total weight of low-substituted hydroxypropyl cellulose, mix well, then add 19% of the total weight of vinegar as a wetting agent, mix well, make pills by plastic method, pass through a pill making machine, make 1000g pills, vacuum dry at 80°C for 16 hours, and get the finished product.

[0042] Dosage and Administration: Take the medicine three times a day, 20 pills each time.

[0043] Example 4.

[0044] Prescription: 8g of Pseudostellaria heterophylla, 10g of dried tangerine peel, 2g of Acorus calamus, 9g of Gynostemma pentaphyllum, and 6g of Oxalis sorrel.

[0045] Process: Weigh all the medicinal materials in the prescription according to the proportion, add 8 times the amount of aqueous solution, reflux extraction twice, each time for 1.5 hours, filter, combine the filtrate, concentrate the filtrate to a relative density of 1.22, vacuum dry at 90°C to form a dry extract, grind into fine powder, add 1.0 times the soluble starch, mix, add 3% of the total weight of low-substituted hydroxypropyl cellulose, mix well, then add 18% of the total weight of vinegar as a wetting agent, mix well, make pills by plastic method, pass through a pill making machine, make 1000g pills, vacuum dry at 70°C for 15 hours, and get the finished product.

[0046] Dosage and Administration: Take the medicine three times a day, 20 pills each time.

[0047] Example 5.

[0048] Prescription: 8g of Pseudostellaria heterophylla, 2g of dried tangerine peel, 5g of Acorus calamus, 8g of Gynostemma pentaphyllum, and 10g of Oxalis sorrel.

[0049] Process: Weigh all the medicinal materials in the prescription according to the proportion, add 10 times the amount of aqueous solution, reflux extraction twice, 1 hour each time, filter, combine the filtrate, concentrate the filtrate to a relative density of 1.24, vacuum dry at 70°C to form a dry extract, grind into fine powder, add 1.2 times the soluble starch, mix, add 4% of the total weight of low-substituted hydroxypropyl cellulose, mix well, then add 16% of the total weight of vinegar as a wetting agent, mix well, make pills by plastic method, pass through a pill making machine, make 1000g pills, vacuum dry at 60°C for 13 hours, and get the finished product.

[0050] Dosage and Administration: Take the medicine three times a day, 20 pills each time.

[0051] The present invention has carried out a lot of experimental studies, and the following are the results of the experimental studies of the present invention:

[0052] 1. Preparation process test

[0053] 1.1 Instruments and Materials

[0054] 1.1.1 Instruments: high performance liquid chromatograph (Shimadzu, LC-2030), AG-135 electronic balance; ultrapure water instrument; ultrasonic cleaning instrument; temperature-controllable electric heating jacket (KDM type; Shandong Chenghualu Electric Heating Instrument Co., Ltd.); rotary evaporator; electric constant temperature water bath (XMTD-4000 type), electronic balance [(SI-234; Denver Instrument Co., Ltd. (0.1 mg)]; low-temperature coolant circulation pump.

[0055] 1.1.2 Materials

[0056] 1.1.2.1 Medicinal pieces: Radix Glycyrrhizae Preparata, Radix Pseudostellariae, Pericarpium Citri Reticulatae, Oxalis Sorrel, and Acorus Tatarinowii.

[0057] 1.1.2.2 Reference reagents: Hesperidin reference substance (Batch No.: 110721-201818). Except for acetonitrile which is of chromatography grade, all other reagents are of chromatography grade.

[0058] 1.3 Research on extraction process

[0059] 1.3.1 Research on extraction process

[0060] According to the physicochemical properties of the medicinal pieces, the reflux method was used for extraction. The content of hesperidin, the extract yield, and the small intestine propulsion rate were determined as evaluation indicators. According to the proportion of each medicine in the prescription and the monarch, minister, adjuvant, and guiding medicine, the total weight coefficient was set to 100, of which the weight coefficients of hesperidin content, extract yield, and small intestine propulsion rate accounted for 20, 30, and 50, respectively.

[0061] Hesperidin content score = (hesperidin content / maximum hesperidin content) × 20

[0062] Extract yield score = (extract yield / maximum extract yield) × 30

[0063] Small intestinal propulsion rate score = (carbon end length / total small intestinal length) × 50

[0064] Comprehensive score = hesperidin content score + extract yield score + small intestine propulsion rate score

[0065] 1.3.1.1 Investigation of water absorption

[0066] During the process of adding liquid for extraction, the medicinal slices will absorb part of the volume of liquid, resulting in the loss of the extract and affecting the accuracy of the experimental results. Therefore, it is necessary to examine the water absorption rate of the medicinal slices and replenish the water absorption during the extraction process to ensure the accuracy of the experimental results.

[0067] Weigh the water-extracted medicinal material slices, add 10 times the amount of water to soak until the slices become soft and transparent, filter and weigh, discard the remaining water, and calculate the water absorption rate. The results are shown in Table 1-1. Figure 2 .

[0068] Water absorption rate = [(weight of the slice after water absorption - weight of the slice before water absorption) / weight of the slice before water absorption] × 100%.

[0069] Table 2-1 Investigation of water absorption rate of Chinese herbal medicine pieces

[0070]

[0071] From Table 1-1, Figure 2 The experimental results show that after the medicinal materials are soaked for 100 minutes, the water absorption is basically stable, and the medicinal materials are soaked to the core, indicating that the water absorption of the medicinal materials has reached saturation at 100 minutes. At this time, the average water absorption of the three medicinal materials is 157.37% of the medicinal materials. Based on actual production, in order to fully soak the medicinal materials, the medicinal materials soaking time is selected as 100 minutes.

[0072] 1.3.1.2 Investigation of extract yield: Place 20 ml of the test solution in a dried constant-weight evaporating dish, evaporate to dryness in a water bath, dry in an oven (105°C) for 3 h, take out, cool in a desiccator and weigh, obtain the weight of the residue, and calculate the extract yield.

[0073]

[0074] 1.3.1.3 Determination of hesperidin content in water extract

[0075] 1.3.1.3.1 Chromatographic conditions: Chromatographic column: Symmetry C18 (4.6 mm × 150 mm, 5 μm); acetonitrile-water (22:78) as mobile phase; volume flow rate: 1 ml / min; injection volume: 5 μL; detection wavelength: 283 nm. Column temperature: 30°C.

[0076] 1.3.1.3.2 Preparation of reference solution: Take an appropriate amount of hesperidin reference substance, weigh it accurately, and add methanol to make a solution containing 0.4 mg per 1 ml.

[0077] 1.3.1.3.3 Preparation of test solution: Grind the product into powder, accurately weigh 0.5 g, place in a 250 ml conical flask with a stopper, accurately add 25 ml of methanol, weigh the weight (accurate to 0.01 g), ultrasonicate in an ultrasonic machine for 45 min, cool, make up the lost weight with the extract, shake well, filter with filter paper, filter with a microporous filter membrane (0.22 μm), and take the filtrate to obtain the solution.

[0078] 1.3.1.3.4 Determination method: Accurately pipette 5 μL of reference solution and test solution respectively, inject into liquid chromatograph to determine the peak area and calculate the content of hesperidin.

[0079] 3.1.4 Pharmacological index determination and methods

[0080] 3.1.4.1 Small Intestine Propulsion Rate Test: The drug-administered groups were all given the test substance 0.2 mL / 10 g by oral gavage, once a day, for 7 consecutive days. Before the last administration, the animals in each group were fasted but not watered for 24 h. 30 min after the last administration, the mice in each group were given charcoal powder by gavage. 20 min after the administration of charcoal powder, the animals were killed by dislocation of the cervical vertebrae, the abdominal cavity was opened to separate the mesentery, the intestinal tube from the pylorus at the upper end and the ileocecal part at the lower end was cut, and placed on a tray. The small intestine was gently pulled into a straight line, and the intestinal length was measured as the "total length of the small intestine", and the length from the pylorus to the front of the charcoal powder was the "charcoal powder propulsion length", and the charcoal powder propulsion rate was calculated.

[0081] Charcoal powder advancement rate = (charcoal powder advancement length (cm) / total length of small intestine (cm) × 100%

[0082] 1.3.2 Single Factor Experiment

[0083] Single factor tests were conducted on extraction time, water addition amount, soaking time and extraction times, and the level of each influencing factor was determined based on the test results.

[0084] 1.3.2.1 Choice of water addition amount: Take 10 times the amount of medicinal materials in the prescription, and measure the average water absorption of the medicinal materials to be 1.56 times, so choose to add water amount of 6, 8, or 10 times the amount of the medicinal materials.

[0085] 1.3.2.2 Selection of decoction times: According to the conventional method, the extraction times are selected as follows: once, twice, or three times.

[0086] 1.3.2.3 Selection of decoction time: By consulting relevant literature and based on the properties of the drugs in the prescription, the decoction times were selected as: 1.0, 1.5, and 2.0 hours, respectively.

[0087] 1.3.3 Orthogonal test method for optimal extraction process

[0088] 1.3.3.1 Based on the results of the single-factor experiment, determine the level of each influencing factor.

[0089] Orthogonal design was carried out with extraction time (A), number of extractions (B), and water addition amount (C) as evaluation indicators. The experimental factors and levels are shown in Tables 1-2 and 1-3; the orthogonal test results and variance analysis are shown in Tables 1-4 and 2-5.

[0090] Table 1-2 Extraction process orthogonal test factor level table

[0091]

[0092] Table 1-3 Orthogonal test table of extraction process

[0093] Experiment No. A B C D 1 1 1 6 1 2 1 2 8 2 3 1 3 10 3 4 1.5 1 8 3 5 1.5 2 10 1 6 1.5 3 6 2 7 2 1 10 2 8 2 2 6 3 9 2 3 8 1

[0094] 1.3.3.2 Orthogonal test results

[0095] Table 2-4 Orthogonal experiment table of extraction process and experimental results table

[0096]

[0097] Table 1-5 Variance analysis table

[0098]

[0099]

[0100] Note: F 0.05 (2, 2) = 19. F 0.01 (2,2)=99.00. *P<0.05.

[0101] 1.3.3.3 Results and Analysis

[0102] From the results in Tables 1-4, it can be seen that the best extraction process of the present invention is A3B3C3, that is, extraction for 2 hours, extraction for 3 times, and adding 10 times the amount of aqueous solution. From the range R, it can be seen that the number of extractions (B) has the greatest impact on the extraction, followed by the amount of water added (C), and finally the extraction time (A). The variance is estimated with the D factor as the error term.

[0103] It can be seen from Tables 1-5 that the number of extractions (B) has a significant influence on the extraction effect, and the influence is relatively large. Although the extraction time (A) and the amount of water added (C) have significant effects, as can be seen from Tables 2-4, the difference is not very large. Taking into account the factors of saving time and cost, the optimal extraction process is determined to be (A1B3C3), that is, extraction for 1h, extraction 3 times, and adding 10 times the amount of aqueous solution.

[0104] 1.3.3.4 Confirmatory test: According to the determined optimal process, three parallel confirmation tests were carried out to measure the hesperidin content, the paste yield and the small intestine propulsion rate respectively. The results are shown in Table 1-6. The results show that the process is reasonable, stable and feasible.

[0105] Table 1-6 Extraction process verification results

[0106]

[0107] 1.4 Investigation of concentration and drying process

[0108] 1.4.1 Investigation of the Extract Concentration Process

[0109] Concentration is an important process step in the preparation of pills. The commonly used concentration methods in production are atmospheric pressure concentration and vacuum concentration. Atmospheric pressure concentration takes a long time, has a high concentration temperature, and a slow concentration speed; vacuum concentration has the advantages of low temperature and preventing or reducing the decomposition of heat-sensitive components. This experiment used hesperidin content as an indicator to investigate vacuum concentration and atmospheric pressure concentration of the extract. Two temperatures of 65°C and 85°C were set for vacuum concentration, and one portion was concentrated at atmospheric pressure at 100°C to select the best concentration process for the extract.

[0110] 1.4.1.1 Test method: Weigh 1 portion of 10 times the prescription medicinal material, extract according to the best extraction process, and obtain the extract. The extract is divided into 3 portions, and concentrated at 65℃, 85℃, and 100℃ respectively, and concentrated to about 150ml. Accurately pipette 5mL of the concentrate into an evaporating dish, take two portions at each temperature, evaporate in a water bath (100℃), add methanol to fully dissolve in a 100mL volumetric flask, add methanol to the scale, shake well, pass through a 0.22μm microporous filter membrane, and take the filtrate. Determine the hesperidin content according to the conditions under 1.3.1.3. The results are shown in Table 1-7.

[0111] Table 1-7 Results of investigation on extract concentration method and temperature

[0112]

[0113] Result analysis: The hesperidin content was the highest under 100℃ atmospheric pressure concentration conditions than under 65℃ and 85℃ reduced pressure concentration conditions. In order to save the concentration time and improve the concentration efficiency, the extract was concentrated at 100℃ atmospheric pressure.

[0114] 1.5 Investigation of concentrated extract drying process

[0115] 1.5.1 Drying method and temperature investigation

[0116] Common drying methods used in production include normal pressure drying and reduced pressure drying. Although normal pressure drying takes a long time and has a high drying temperature, it is still used in production; reduced pressure drying has the advantages of low temperature, fast drying speed, and easy collection and crushing of the dried product; this experiment used the content of hesperidin as an indicator to investigate the drying method of the extract, set up normal pressure drying and reduced pressure drying, and selected the best drying method for concentrated extract.

[0117] 1.5.1.1 Test method: Weigh 1 / 2 of the prescription medicinal materials, 18 portions, extract according to the best extraction process to obtain the extract, concentrate the extract to a relative density of 1.22 to 1.24 at normal pressure, pour it into a square plate covered with plastic film, and conduct two parallel portions for each test method. Six portions are subjected to forced air drying (60°C, 80°C, 100°C), and six portions are subjected to vacuum reduced pressure drying (60°C, 70°C, 80°C, -0.06 to -0.08MPa). 6 portions were subjected to microwave vacuum drying (300W, 500W, 1000W, -0.00~0.1MPa), and 1.0g of dry extract was accurately weighed respectively, placed in a stoppered conical flask, 25mL of methanol was accurately added, the flask was sealed, the weight was determined, ultrasonic treatment (power 300W; frequency 40kHZ) was performed for 45 minutes, the flask was cooled, the weight was determined again, the weight loss was supplemented with methanol, the filtrate was shaken, the filtrate was filtered through a 0.22μm microporous membrane, and the hesperidin content was determined according to the conditions under the content determination item, and the results are shown in Table 1-8.

[0118] Table 1-8 Results of investigation on concentrated extract drying methods

[0119]

[0120] 1.6 Results: The concentration and drying process is as follows: the extract is concentrated under normal pressure, and the concentrated extract is dried under vacuum. The hesperidin content is highest when the drying temperature is 80°C.

[0121] 1.7 Molding process research

[0122] 1.7.1 Screening of single excipients: Weigh the dry extract of Example 1 and shape it according to the ratio of powder to excipients in the table, with each powder being 40g. Observe the quality of the soft material and evaluate the pellets (see Table 1-9).

[0123] Table 1-9 Single auxiliary material soft material quality and pelleting evaluation

[0124]

[0125]

[0126] 1.7.2 Screening of mixed excipients: Single excipients cannot meet the requirements of pill making, so mixed excipients are screened. Weigh the dry extract obtained by the best extraction process and shape it according to the ratio of paste powder to excipients in the table. Each paste powder is 40g. The quality of soft materials and pill evaluation are shown in Table 1-10.

[0127] Table 1-10 Quality of mixed auxiliary materials and pelleting evaluation

[0128]

[0129] Since adding a small amount of disintegrant can improve the quality of the soft material, low-substituted hydroxypropyl cellulose was selected for the test. Its common dosage is 2%-5%. Take 40g of extract powder, add 1.1 times the weight of starch of the extract powder and 2.2% low-substituted hydroxypropyl cellulose of the total weight of the extract powder and starch as disintegrant, add 17% vinegar solution, and the quality of the soft material and the evaluation of the pellets are shown in Table 1-11.

[0130] Table 1-11 Evaluation of soft material quality and pelleting after adding disintegrant

[0131]

[0132] 1.7.3 Results: The optimal molding process is: dry extract powder, add 1.1 times starch, mix thoroughly, use 2.2% low-substituted hydroxypropyl cellulose as a disintegrant based on the total amount of extract powder and starch, mix thoroughly, add 17% vinegar solution as a wetting agent, and make a soft material with moderate hardness and hardness, and make pills. The pellets have uniform color and good roundness.

[0133] 1.7.4 Moisture content under different drying methods, temperatures and times

[0134] In order to meet the relevant provisions of the moisture content in the 0108 pills item of the fourth general rule of the 2020 edition of the Chinese Pharmacopoeia, further drying is required. Drying to a moisture content of less than 9.0%, the results are shown in Table 1-12.

[0135] Table 1-12 Moisture investigation results of different drying methods, temperatures and times

[0136]

[0137]

[0138] Result analysis: The 2020 edition of the Chinese Pharmacopoeia stipulates that the moisture content of the pills must be less than 9%, and the most appropriate range is to keep the moisture content at around 7%. From the above results, it can be seen that the most suitable moisture content at 65°C under normal pressure is 46 hours of drying, and at 80°C it is 18 hours of drying. The most suitable moisture content at 65°C under vacuum is 14 hours of drying, and at 80°C it is 8 hours.

[0139] 1.7.5 Investigation of the best drying method and temperature: To further select the best drying method and temperature, taking the hesperidin content as an indicator, the pellets dried at 65°C at normal pressure for 46 h, 80°C for 18 h, and the pellets dried at 65°C in vacuum for 14 h, and 80°C for 8 h were ground into powder. Two parallel batches were made for each condition. The test samples were prepared and the hesperidin content was determined according to the pharmacopoeia method. The results are shown in Table 2-13.

[0140] Table 2-13 Best drying method and temperature investigation results

[0141]

[0142] Result analysis: The results show that the best condition is vacuum drying at 65℃.

[0143] 1.7.6 Investigation of the optimal drying time: Grind the pellets dried at 65°C under vacuum for 10h, 14h, and 22h (with a slightly larger moisture interval). For each condition, make two parallel batches and accurately weigh 0.2g of pellet powder from each batch. Prepare the test samples and determine the hesperidin content according to the pharmacopoeia method. The results are shown in Table 1-14.

[0144] Table 1-14 Hesperidin content determination results

[0145]

[0146] 1.8 Results: The hesperidin content of the pills dried at 65℃ for 14h under vacuum conditions was higher, so the best drying time was 14h. Therefore, the best drying condition was: vacuum drying at 65℃ for 14h.

[0147] 1.9 Forming process verification test: According to the best forming process obtained, 3 batches of samples were prepared, and the moisture content, dissolution time and weight difference were investigated according to the "General 0108" of the 2020 edition of the "Pharmacopoeia of the People's Republic of China: Part IV". The results showed that the pellets obtained from the 3 experiments were round and smooth, with uniform color and size, and the moisture content and dissolution time were in compliance with the regulations, and RSD <5%. The results are shown in Table 13. It was obtained that the dry extract was vacuum dried (80°C), crushed into fine powder, 1.1 times starch was added, and after fully mixing, the paste powder and 2.2% low-substituted hydroxypropyl cellulose of the total amount of starch were used as disintegrants. After fully mixing, 17% vinegar solution was added as a wetting agent. The pellets were uniform in color and had good roundness. The pills obtained by vacuum drying at 65°C for 14 hours had reasonable, stable and reproducible effects.

[0148] Table 1-13 Quality evaluation results of 3 batches of samples

[0149]

[0150] 1.10 Content determination

[0151] 1.10.1 Determination of content of raw medicinal materials

[0152] Take about 0.2g of dried tangerine peel and prepare the test solution according to the test sample processing method under the dried tangerine peel item in the 2020 edition of the Chinese Pharmacopoeia. Take 5μL and inject it into the liquid chromatograph. The results are shown in Table 1-14.

[0153] Table 1-14 Determination results of hesperidin content in tangerine peel raw materials

[0154]

[0155] Result analysis: The standard stipulates that the hesperidin content in dried tangerine peel should not be less than 3.5%. The test results are in line with the content regulations under dried tangerine peel in the Chinese Pharmacopoeia.

[0156] 1.10.2 Determination of content of finished products

[0157] Accurately weigh the batch sample prepared in "Item 1.9", inject 5 μL into the HPLC, and determine its hesperidin content under the above conditions. The results are shown in Table 1-15.

[0158] Table 1-15 Hesperidin content determination results

[0159]

[0160]

[0161] From the above sample test results, it can be seen that the hesperidin transfer rate is 21.135%. According to the content limit specified in the Chinese Pharmacopoeia under the item of dried tangerine peel, the hesperidin content limit per gram of preparation is calculated as follows: Hesperidin content limit = (preparation containing dried tangerine peel medicinal material (g) × dried tangerine peel medicinal material containing hesperidin limit × transfer rate / preparation amount (g)) = (297 × 3.5% × 21.315% / 1000) = 0.0022mg / g. Considering the influence of a series of processes such as processing, preparation, preparation, and production of medicinal materials, the product content limit is reduced by 20%. It is tentatively determined that the hesperidin content per gram of this product shall not be less than 0.00044mg.

[0162] 2. Pharmacological Effects Research

[0163] 2.1 Materials

[0164] 2.1.1 Experimental animals: 154 clean-grade healthy Kunming mice of both sexes, weighing 18-22 g, provided by Hunan Changsha Tianqin Experimental Animal Co., Ltd., license number: SCXK(Xiang)2022-0011.

[0165] 2.1.2 Drugs and preparation: The pills prepared according to Example 1 were crushed into powder, enteric-coated aspirin tablets (batch number: 22040201; manufacturer: Shenyang Aojina Pharmaceutical Co., Ltd.), compound berberine tablets (batch number: 210903; manufacturer: Hubei Norde Shengli Co., Ltd.), loperamide hydrochloride capsules (batch number: LLJ0834; manufacturer: Xi'an Janssen Pharmaceutical Co., Ltd.), atropine sulfate tablets (batch number: T220038; manufacturer: Hangzhou Minsheng Pharmaceutical Co., Ltd.), compound honeysuckle granules (batch number: 20220523; manufacturer: Heilongjiang Wusulijiang Pharmaceutical Co., Ltd. Harbin Branch), hemp seed pills (batch number: 210510; manufacturer: Nanjing Tongrentang Pharmaceutical Co., Ltd.), purified water, 0.6% acetic acid solution, raw rhubarb extract, castor oil, and charcoal suspension.

[0166] 2.1.3 Reagents: Mouse 5-HT ELISA KIT, catalog number: ZC-37715W, specification: 48 Test, produced by Shanghai Zhuocai Biotechnology Co., Ltd.; Mouse Camp ELISA KIT, catalog number: ZC-38246W, specification: 48 Test, produced by Shanghai Zhuocai Biotechnology Co., Ltd.; Mouse PGE2 ELISA KIT, catalog number: ZC-38572W, specification: 48 Test, produced by Shanghai Zhuocai Biotechnology Co., Ltd.; Mouse SP ELISA KIT, catalog number: ZC-37822W, specification: 48 Test, produced by Shanghai Zhuocai Biotechnology Co., Ltd.; Mouse β-EP ELISA KIT, catalog number: ZC-37893W, specification: 48 Test, produced by Shanghai Zhuocai Biotechnology Co., Ltd.

[0167] 2.1.4 Instruments and consumables: ELISA reader, model, SpectraMAX Plus384, produced by Meigu Molecular Instrument Co., Ltd.; UPH-Ⅱ-10T ultrapure water manufacturing system, model, UPH-Ⅱ-10T, produced by Chengdu Ultrapure Technology Co., Ltd.; electronic constant temperature water bath, model DZKW-4, produced by Beijing Zhongxing Weiye Instrument Co., Ltd.; pipettes, specifications, 1000μL, 200μL, 50μL, produced by Dalong Xingchuang Experimental Instrument (Beijing) Co., Ltd.; gun tips, specifications, 1000μL, 200μL, 50μL, produced by biosharp.

[0168] 2.2 Methods

[0169] 2.2.1 Effects on castor oil-induced diarrhea in mice

[0170] 84 km mice, weighing 18-22 g, were randomly divided into 7 groups, 12 mice in each group. They were blank group, model group, positive drug group 1 (compound berberine tablet group, 0.15 g kg-1 ·d -1 ), positive drug group 2 (loperamide hydrochloride capsule group, 1 mg·kg -1 ·d -1 ); The pills prepared in Example 1 were crushed into powder and grouped according to the dosage to obtain a low-dose group of pills (1.48 mg·kg -1 ·d -1 ), medium-dose pill group (2.96 mg kg -1 ·d -1 ), high-dose pill group (5.92 mg kg -1 ·d -1 ). The blank group and the model group were gavaged with the same volume of pure water for 7 consecutive days. 30 minutes after the last administration, all groups except the blank group were gavaged with 0.2 ml / 10 g of castor oil, and each mouse was placed in a clean metabolic cage with clean filter paper on the bottom of the cage, and the feces discharge within 4 hours was counted.

[0171] Diarrhea rate (%) = (number of mice with loose stools / total number of mice in the group) × 100%; Diarrhea index = loose stool rate × loose stool grade; loose stool rate (%) = (number of loose stools excreted by mice / total number of stools) × 100%; loose stool grade = sum of loose stool stain grades / number of loose stools. If there are stains on the filter paper around the feces, it is loose stool, otherwise it is dry stool.

[0172] Method for determining the level of loose stool stains: Use a ruler to measure the diameter of the stains. If the stains are not circular, take the average value of the sum of the lengths of the long and short diameters, i.e., level 1 is less than 1 cm, level 2 is 1-2 cm, level 3 is 2.1-3 cm, and level 4 is 3.1-4 cm. Calculate the sum of the loose stool stain levels for each mouse, then add up all the loose stool levels of the mouse, divide by the number of loose stools, and get the average loose stool level.

[0173] 2.2.2 Effect of pills on rhubarb-induced diarrhea in mice

[0174] This part is the same as the experimental animal grouping and dosing under "1.2.1". 30 minutes after the last dosing, except for the blank group, each group was gavaged with 100% rhubarb solution 0.4ml / 10g. Each mouse was placed in a clean metabolic cage with clean filter paper on the bottom of the cage, and the feces discharge within 4 hours was counted.

[0175] 2.2.3 Effect of pills on acetic acid-induced writhing reaction in mice

[0176] 2.2.3.1 Trial Grouping and Dosing

[0177] A total of 70 km mice were randomly divided into 7 groups, namely, blank group, model group, positive drug group 1 (aspirin enteric-coated tablets group, 200 mg kg -1 ·d -1), positive drug group 2 (compound honeysuckle granules group, 12 g·kg-1·d-1), low-dose pill group (1.48 mg·kg -1 ·d -1 ), medium-dose pill group (2.96 mg kg -1 ·d -1 ), high-dose pill group (5.92 mg kg -1 ·d -1 ); the blank group and the model group were gavaged with the same volume of pure water for 7 consecutive days. 60 minutes after the last administration, each group was intraperitoneally injected with 0.6% acetic acid solution 0.2ml / 10g, and the timing was started after the injection. The number of twists within 15 minutes after the first twist was recorded, and the number of twists and the inhibition rate of twisting reaction were calculated.

[0178] Observation criteria: abdominal cavity concavity, extension of hind limbs, and lifting of buttocks (when all the above behaviors occur, it is counted as one writhing reaction). Writhing reaction inhibition rate (%) = (average number of writhing reactions in the model group - average number of writhing reactions in each drug-administered group) / average number of writhing reactions in the model group × 100%.

[0179] 2.2.3.2 Serum β-EP and PGE2, cAMP, serum 5-hydroxytryptamine (5-HT), and SP levels

[0180] The number of twisting times of mice in the analgesia experiment was recorded within 15 minutes, and then the eyeballs were removed and blood was collected in a 1.5 mL centrifuge tube. The blood was naturally coagulated for 20 minutes at room temperature, centrifuged at 3000 r·min-1 for 20 minutes, and the supernatant was collected and stored at -20℃ for later use. The ELISA method was used to determine the content of serum β-EP and PGE2, cAMP, and serum 5-hydroxytryptamine (5-HT); the specific steps were carried out according to the instructions of the kit.

[0181] 2.2.4 Effect of pills on the propulsion of charcoal powder in the small intestine of mice

[0182] A total of 70 km mice were randomly divided into 7 groups, namely, blank group, model group, positive drug group 1 (atropine sulfate group, 0.3 mg·kg -1 ·d -1 ), positive drug group 2 (Ma Ren Wan group, 1 g·kg -1 ·d -1 ), low-dose pill group (1.48 mg kg -1 ·d -1 ), medium-dose pill group (2.96 mg kg -1 ·d -1 ), high-dose pill group (5.92 mg kg -1 ·d -1); the blank group and the model group were gavaged with the same volume of pure water for 7 consecutive days. Before the last administration, the animals in each group were fasted but not watered for 24 hours. The last administration of drug powder and charcoal suspension was performed, and the charcoal concentration was 1%. After 20 minutes, the animals were killed by dislocating the cervical vertebrae, the abdominal cavity was opened to separate the mesentery, and the intestinal tube from the pylorus at the upper end and the ileocecal part at the lower end was cut and placed on a tray. The small intestine was gently pulled into a straight line, and the length of the intestinal tube was measured as the "total length of the small intestine", and the length from the pylorus to the front of the charcoal was the "charcoal advancement length", and the charcoal advancement rate was calculated.

[0183] Charcoal powder advancement rate = charcoal powder advancement length (cm) / total small intestine length (cm) × 100%.

[0184] 2.2.5 Statistical methods

[0185] The experimental data were analyzed using statistical software SPSS 26.0, and all data were expressed as x±s. The data were first tested for normality and homogeneity of variance. The data were in accordance with normal distribution and homogeneity of variance. One-way ANOVA was used, and the least significant difference (LSD) was used for pairwise comparison between groups. P<0.05 was considered statistically significant.

[0186] 2.3 Results

[0187] 2.3.1 Effect of pills on castor oil-induced diarrhea in mice

[0188] Compared with the model group, the compound berberine group, loperamide hydrochloride group, and low-, medium-, and high-dose pill groups could significantly reduce the frequency of diarrhea caused by castor oil in mice and alleviate the degree of diarrhea; the loose stool rate, loose stool grade, and diarrhea index were all significantly reduced, with significant differences (P<0.01), as shown in Table 2-1.

[0189] Table 2-1 Effect of castor oil on diarrhea in mice (x±s, n=10)

[0190]

[0191] Note: Compared with the model group, *P<0.05, **P<0.01.

[0192] 2.3.2 Effect of pills on rhubarb-induced diarrhea in mice

[0193] Compared with the model group, the compound berberine group, loperamide hydrochloride group, and the medium and high dose pill groups can significantly reduce the frequency of diarrhea caused by rhubarb in mice and alleviate the degree of diarrhea; the loose stool rate, loose stool grade, and diarrhea index are significantly reduced, with significant differences (P<0.01), but the loose stool grade of the low dose pill group did not change significantly compared with the model group, and the difference was insignificant (P>0.05). See Table 2-2.

[0194] Table 2-2 Effect of pills on diarrhea in mice caused by rhubarb (x±s, n=10)

[0195]

[0196]

[0197] Note: Compared with the model group, *P<0.05, **P<0.01.

[0198] 2.3.3 Effects on acetic acid-induced writhing in mice

[0199] 2.3.3.1 Results of the acetic acid writhing test

[0200] Compared with the model group, the number of acetic acid writhing reactions in mice in the aspirin group, compound honeysuckle granule group, and low-dose, medium-dose, and high-dose pill groups was significantly reduced, and the pain reaction of mice induced by acetic acid was significantly inhibited (P<0.01). The differences were statistically significant, as shown in Table 2-3.

[0201] Table 2-3 Experimental results of the number of writhing times and inhibition rate of acetic acid writhing mice in response to pain (x±s, n=10)

[0202]

[0203] Note: Compared with the model group, *P<0.05, **P<0.01.

[0204] 2.3.3.2 Serum β-EP and PGE2, cAMP, serum 5-hydroxytryptamine (5-HT), and SP content

[0205] Compared with the model group, the serum 5-HT concentration of mice in the compound honeysuckle granules group and the high-dose pill group was significantly reduced (P<0.05); the serum 5-HT concentration of mice in the aspirin tablet group and the low- and medium-dose pill groups was not significantly different. Compared with the blank group, the serum 5-HT concentration of mice in the model group was significantly increased, with a very significant difference (P<0.01). The results are shown in Table 2-4.

[0206] Table 2-4 Changes in serum 5-HT concentration in each group of mice (x±s , n=10)

[0207]

[0208] Note: Compared with the blank group, * P<0.05, ** P<0.01; compared with the model group, # P<0.05, ## P<0.01.

[0209] Compared with the blank group, the cAMP concentration in the serum of the mice in the model group was significantly increased, and the difference was extremely significant (P

[0210] <0.01). Compared with the model group, the cAMP concentration in the serum of mice in the high-dose pill group was significantly reduced, with a very significant difference (P<0.01); the cAMP concentration in the serum of mice in the aspirin tablet group and the compound honeysuckle granule group was significantly reduced (P<0.05); the cAMP concentration in the serum of mice in the low- and medium-dose pill groups was not significantly different. The results are shown in Table 4-5.

[0211] Table 1-5 Changes in serum cAMP concentration in each group of mice (x±s, n=10)

[0212]

[0213] Note: Compared with the blank group, * P<0.05, ** P<0.01; compared with the model group, # P<0.05, ## P<0.01.

[0214] Compared with the blank group, the PGE2 concentration in the serum of the mice in the model group was significantly increased, and the difference was extremely significant (P

[0215] <0.01). Compared with the model group, the serum PGE2 concentration of mice in the compound honeysuckle granules group and the high-dose pill group was significantly reduced, with extremely significant differences (P<0.01); the serum PGE2 concentration of mice in the aspirin tablets group and the medium-dose pill group was significantly reduced, with significant differences (P<0.05); the serum PGE2 concentration of mice in the low-dose pill group was not significantly different, as shown in Table 2-6.

[0216] Table 2-6 Changes in serum PGE2 concentrations in each group of mice (x±s, n=10)

[0217]

[0218] Note: Compared with the blank group, * P<0.05, ** P<0.01; compared with the model group, # P<0.05, ## P<0.01.

[0219] Compared with the blank group, the concentration of SP in the serum of mice in the model group was significantly increased, and the difference was extremely significant (P<0.01).

[0220] Compared with the model group, the SP concentration in the serum of mice in the compound honeysuckle granule group was significantly reduced, and the difference was extremely significant (P<0.01); the SP concentration in the serum of mice in the aspirin tablet group and the high-dose pill group was reduced, and the difference was significant (P<0.05); there was no significant difference in the SP concentration in the serum of mice in the low-dose and medium-dose pill groups. The results are shown in Table 2-7.

[0221] Table 2-7 Changes in SP concentration in serum of mice in each group (x±s, n=10)

[0222]

[0223] Note: Compared with the blank group, * P<0.05, ** P<0.01; compared with the model group, # P<0.05, ## P<0.01.

[0224] Compared with the blank group, the serum β-EP concentration of the model group mice was significantly increased, and the difference was extremely significant (P

[0225] <0.01). Compared with the model group, the serum β-EP concentrations of mice in the medium and high dose pill groups were significantly reduced, with extremely significant differences (P<0.01); the serum β-EP concentrations of mice in the aspirin tablet group and the compound honeysuckle granule group were significantly reduced (P<0.05); the serum β-EP concentrations of mice in the low dose pill group were not significantly different. The results are shown in Table 2-8.

[0226] Table 2-8 Changes in β-EP concentration in serum of mice in each group (x±s, n=10)

[0227]

[0228] Note: Compared with the blank group, * P<0.05, ** P<0.01; compared with the model group, # P<0.05, ## P<0.01.

[0229] 2.3.4 Effects of charcoal powder propulsion in the small intestine of mice

[0230] Compared with the model group, the charcoal powder advancement length and small intestinal advancement rate of mice in the Ma Ren Pill group, medium and high dose pill groups were significantly increased, and the difference was statistically significant (P<0.05). Among them, compared with the model group, the charcoal powder advancement length and small intestinal advancement rate of mice in the high dose pill group were extremely significant (P<0.01). Compared with the model group, there was no significant change in the charcoal powder advancement length and small intestinal advancement rate of mice in the atropine sulfate group and low dose pill group, and the difference was not statistically significant (P>0.05). The results are shown in Table 2-9.

[0231] Table 2-9 Experimental results of the effect of pills on the propulsion of charcoal powder in the small intestine of mice (x±s, n=10)

[0232] Grouping <![CDATA[Total length of small intestine / cm > <![CDATA[Carbon residue propulsion length / cm > Carbon powder advancement rate / 100% Model Group 46.79±4.05 24.88±3.09 0.53±0.06 Atropine sulfate group 48.03±3.80 24.00±5.77 0.50±0.10 Ma Ren Wan Group 49.17±6.09 31.89±4.30* 0.65±0.10* Low-dose pill group 46.58±4.85 27.00±6.07 0.58±0.12 Medium dose pill group 48.83±3.90 31.67±5.42* 0.65±0.13* High-dose pill group 47.96±5.01 38.17±4.18** 0.80±0.06**

[0233] Note: Compared with the model group, *p<0.05, **p<0.01.

[0234] 2.4 Discussion and Summary

[0235] In the above efficacy test, the effect of the present invention on the diarrhea of ​​mice caused by rhubarb and castor oil is to investigate the antidiarrheal effect of the present invention. The results show that each dosage group of the pill of the present invention has a significant inhibitory effect on acetic acid writhing.

[0236] The acetic acid writhing method investigates the analgesic effect of the pills of the present invention. The acetic acid writhing method is more inclined to the acute inflammatory pain model and is a relatively sensitive and common chemical stimulation pain method for screening analgesics. This method is a chemical pain caused by H+ stimulation, and an inflammatory reaction will also occur at the same time. The non-steroidal compound aspirin has a strong effect on this type of pain. Aspirin is used as a positive control drug for Western medicine in the acetic acid writhing method. At the same time, the compound honeysuckle granules are selected as a positive control of traditional Chinese medicine. It can be seen from the test results of the acetic acid writhing method that each dose group can significantly reduce the number of writhing times, and the effect is obvious. In this study, the ELISA method was used to detect the contents of β-EP and PGE2, cAMP, serum 5-hydroxytryptamine (5-HT), and SP in the serum of mice in the acetic acid writhing experiment. It was found that the medium and high dose groups of the pills had the most significant effect on the downregulation of β-EP and PGE2, cAMP, serum 5-hydroxytryptamine (5-HT), and SP levels in the mouse serum, and exerted anti-inflammatory and analgesic effects. The changes in 5-HT levels in serum and cerebral cortex are consistent with related research reports that 5-HT plays a pain-inducing role in the periphery and is an active analgesic substance in the central nervous system.

[0237] Intestinal propulsion measurement is the most commonly used classical method to investigate gastric and intestinal motility. This study investigated the effect of the present invention on intestinal motility of mice by oral administration to experimental mice. The results showed that: ① The positive drug Ma Ren Wan and the medium and high doses of the pills of the present invention can significantly increase the small intestinal propulsion rate and promote intestinal peristalsis, while atropine sulfate is commonly used as an acetylcholine inhibitor and has a significant inhibitory effect on intestinal peristalsis, which further proves that the medium and high dose groups of the pills of the present invention and the Ma Ren Wan group can significantly promote intestinal peristalsis; based on the intestinal propulsion results, it can be preliminarily inferred that the pills of the present invention may exert their effects on the intestinal segment.

Claims

1. A drug for irritable bowel syndrome, characterized in that: The medicinal effective ingredients are calculated by weight and are prepared from 40-80 parts of Pseudostellaria heterophylla, 40-80 parts of dried orange peel, 40-80 parts of Acorus calamus, 70-110 parts of Cynanchum cyrtonema and 40-80 parts of Oxalis sorrel.

2. The medicament for irritable bowel syndrome according to claim 1, characterized in that: The medicinal effective ingredients are calculated by weight: 60 parts of Pseudostellaria heterophylla, 60 parts of dried orange peel, 60 parts of Acorus calamus, 90 parts of Cynanchum cyrtonema and 60 parts of Oxalis sorrel.

3. The method for preparing a drug for irritable bowel syndrome according to claim 1 or 2, characterized in that: The preparation method is carried out according to the following steps: (1) Weigh all the medicinal materials in the prescription according to the proportion, add 6-12 times the amount of aqueous solution, reflux extraction 1-4 times, each time for 0.5-2h, filter, combine the filtrate, concentrate the filtrate to a relative density of 1.20-1.26, vacuum dry at 60-100℃ to form a dry extract, grind into fine powder, add 0.9-1.3 times the amount of soluble starch, mix, and obtain product A; (2) Weigh 2-5% of the total weight of product A and add low-substituted hydroxypropyl cellulose to product A and mix well to obtain product B; (3) Weigh 10-20% of the total weight of product B as vinegar as a wetting agent, add it to product B and mix well, make pills by plasticizing, pass through a pill making machine, make 1000g pills, and vacuum dry at 50-80℃ for 12-16h to obtain the finished product.

4. The method for preparing a drug for irritable bowel syndrome according to claim 1 or 2, characterized in that: The preparation method is carried out according to the following steps: (1) Weigh all the medicinal materials in the prescription according to the proportion, add 8-12 times the amount of aqueous solution, reflux extraction 2-4 times, each time for 0.5-1.5 hours, filter, combine the filtrate, concentrate the filtrate to a relative density of 1.1-1.3, vacuum dry at 70-90°C to form a dry extract, grind into fine powder, add 1.0-1.5 times the amount of soluble starch, and obtain product A; (2) Weigh 2-5% of the total weight of product A and add low-substituted hydroxypropyl cellulose to product A and mix well to obtain product B; (3) Weigh 10-20% of the total weight of product B as vinegar as a wetting agent, add it to product B and mix well, make pills by plasticizing, pass through a pill making machine, make 1000g pills, and vacuum dry at 60-70℃ for 13-15h to obtain the finished product.

5. The method for preparing a drug for irritable bowel syndrome as claimed in claim 4, characterized in that: The preparation method is carried out according to the following steps: (1) Weigh all the medicinal materials in the prescription according to the proportion, add 10 times the amount of aqueous solution, reflux extraction 3 times, each time for 1 hour, filter, combine the filtrate, concentrate the filtrate under direct fire to a relative density of 1.22~1.24, vacuum dry at 80℃ to form a dry extract, grind into fine powder, add 1.1 times the amount of soluble starch, and obtain product A; (2) Weigh 2.2% of the total weight of product A and add low-substituted hydroxypropyl cellulose to product A and mix well to obtain product B; (3) Weigh 17% of the total weight of product B as vinegar as a wetting agent, add it to product B and mix well, make pills by plasticizing, pass through a pill making machine, make 1000g pills, and vacuum dry at 65℃ for 14h to obtain the finished product.

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