Application of natural benzoic acid compound in preparation of medicine for treating gastrointestinal diseases

Abstract

The invention discloses an application of a natural benzoic acid compound in preparation of a medicine for treating gastrointestinal diseases. According to the invention, a compound 4-hydroxy-3, 5-dimethoxybenzoyl ethyl ester with a remarkable inhibition effect on intestinal smooth muscle contraction is screened from a plurality of natural medicine small molecules, and the compound exists in amorpha fruticosa flower honey and seaweed. Experiments prove that the 4-hydroxy-3, 5-dimethoxybenzoyl ethyl ester has a remarkable curative effect on diarrhea and inflammatory bowel diseases, not only can effectively inhibit intestinal tract movement and reduce diarrhea clinical symptoms such as moisture in excrement, but also can improve the clinical symptoms of colitis such as weight loss, diarrhea and hematochezia of experimental animals; good clinical application prospects are realized in gastrointestinal digestive system diseases; the functions of the small molecules are obviously superior to those of other traditional Chinese medicine natural small molecules with similar structures, and a new thought and a new choice are provided for development of medicines for resisting diarrhea and inflammatory bowel diseases.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly relates to the use of a natural benzoic acid compound in the preparation of a drug for treating gastrointestinal system diseases. Background Art

[0002] Diarrhea is a gastrointestinal disease caused by increased gastrointestinal motility, resulting in an increase in the number of stools and thin, watery feces. It is caused by multiple pathogens and factors. The characteristics of diarrhea are changes in intestinal motility, frequency, and fecal consistency, with an increase in water content, making the feces liquid or pasty. Diarrhea has a high incidence rate, a high recurrence rate after drug withdrawal, and obvious side effects, causing serious harm to the physical and mental health of patients. Currently, the drugs clinically used to treat diarrhea mainly include probiotics, intestinal mucosal protectants, antispasmodics, antibiotics, and traditional Chinese medicine preparations. For a long time, antibiotic drugs have always been the first choice for treating intractable diarrhea clinically, but the disadvantage is that they are prone to addiction, and their application is also restricted accordingly. In recent years, with the in-depth study of traditional medicine, it has been found that it has unique effects in the field of diarrhea.

[0003] Inflammatory bowel disease (IBD) is an idiopathic inflammatory disease involving the ileum, colon, and rectum, with clinical manifestations of repeated diarrhea, abdominal pain, mucus stools, and even bloody stools. IBD includes ulcerative colitis (UC) and Crohn's disease (CD). For a long time, with the changes in people's eating habits and living environment, IBD has gradually evolved into a global disease. The treatment methods of IBD mainly include drug treatment and surgical treatment. Drug treatment mainly includes aminosalicylates, glucocorticoids, biological agents, traditional small molecule immunosuppressants, intestinal antispasmodics, and new small molecule drugs. Currently, the drug treatment of IBD has poor efficacy for a large number of patients, or patients cannot tolerate it due to too many adverse reactions. For refractory diseases, patients urgently need effective alternative solutions.

[0004] New small molecule drugs have the advantages of oral administration, convenient use, stable structure, low manufacturing cost, short half-life, and rapid elimination, showing great application prospects and becoming a research and development hotspot for drugs treating gastrointestinal diseases. Summary of the Invention

[0005] The present invention provides the use of a natural benzoic acid compound in the preparation of a drug for treating gastrointestinal system diseases. The present invention has screened out a small molecule compound 4-hydroxy-3,5-dimethoxybenzoyl ethyl ester from a variety of natural traditional Chinese medicine small molecules, which has a significant inhibitory effect on intestinal smooth muscle contraction and has an obvious effect in anti-diarrhea and inflammatory bowel disease.

[0006] To achieve the above-mentioned invention object, the present invention is implemented by the following technical solutions: The present invention provides an application of a natural benzoic acid compound in the preparation of a drug for treating gastrointestinal diseases.

[0007] Further, the natural benzoic acid compound is ethyl 4-hydroxy-3,5-dimethoxybenzoate, and its structural formula is: 。

[0008] Further, the gastrointestinal diseases include diarrhea and inflammatory bowel disease.

[0009] Further, the dosage of ethyl 4-hydroxy-3,5-dimethoxybenzoate is 10 μM - 300 μM.

[0010] Further, ethyl 4-hydroxy-3,5-dimethoxybenzoate can inhibit the spontaneous contraction of intestinal smooth muscle, inhibit intestinal peristalsis, and reduce the diarrhea index, so as to achieve the effect of treating diarrhea.

[0011] Further, ethyl 4-hydroxy-3,5-dimethoxybenzoate plays a role in relaxing smooth muscle by inhibiting the influx of extracellular calcium.

[0012] Further, ethyl 4-hydroxy-3,5-dimethoxybenzoate can improve the diarrhea, bloody stool symptoms, and colon shortening symptoms of inflammatory bowel disease, reduce the spleen coefficient of patients with inflammatory bowel disease, and the expression of inflammatory factors in the blood.

[0013] The present invention also provides a drug for treating gastrointestinal system diseases, and the active ingredient therein is compound ethyl 4-hydroxy-3,5-dimethoxybenzoate or its stereoisomer, pharmaceutically acceptable salt.

[0014] Further, the drug is a tablet, capsule, dropping pill, granule, injection, powder injection or aerosol.

[0015] Further, the administration methods of the drug include oral administration, intramuscular injection, intravenous injection or infusion, transdermal administration, mucosal administration, aerosol administration.

[0016] Compared with the prior art, the present invention has the following advantages and beneficial effects: The present invention has screened a small molecule compound ethyl 4-hydroxy-3,5-dimethoxybenzoate with a significant inhibitory effect on intestinal smooth muscle contraction from a variety of natural drug small molecules, and for the first time verified through experiments that ethyl 4-hydroxy-3,5-dimethoxybenzoate has a good therapeutic effect in diarrhea and inflammatory bowel disease. This compound has low toxicity, good therapeutic effect, simple structure, and is easy to synthesize.

[0017] The present invention has been verified through in vivo and in vitro experiments. Ethyl 4-hydroxy-3,5-dimethoxybenzoate has a good therapeutic effect on senna-induced osmotic diarrhea, can effectively inhibit the production of loose stools, inhibit intestinal peristalsis, reduce the water content in feces, reduce the number of loose stools, and decrease the diarrhea index. It is a safe and effective drug for treating diarrhea. At the same time, ethyl 4-hydroxy-3,5-dimethoxybenzoate also has a good therapeutic effect on the dextran sulfate sodium-induced murine model of inflammatory bowel disease, can effectively increase the colon length, reduce the disease activity index, and can also improve the clinical symptoms of colitis such as weight loss, diarrhea, and bloody stools in experimental animals, providing new ideas and new options for the development of anti-diarrhea and inflammatory bowel disease drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Shows the effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on the spontaneous contraction of isolated rabbit intestine.

[0019] Figure 2A Shows the effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on acetylcholine-induced intestinal muscle contraction; Figure 2B Shows the effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on histamine-induced intestinal muscle contraction; Figure 2C Shows the effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on barium chloride-induced intestinal muscle contraction; Figure 2D Shows the effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on calcium chloride-induced intestinal muscle contraction.

[0020] Figure 3A Shows the effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on intestinal peristalsis in normal mice; Figure 3B Shows the effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on intestinal peristalsis in diarrhea mice induced by senna.

[0021] Figure 4 Shows the effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on the body weight change of mice with DSS-induced inflammatory bowel disease.

[0022] Figure 5A Shows the effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on the DAI index of mice with DSS-induced inflammatory bowel disease; Figure 5B Shows the effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on the colon length of mice with DSS-induced inflammatory bowel disease.

[0023] Figure 6 Shows the effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on the organ coefficient of mice with DSS-induced inflammatory bowel disease.

[0024] Figure 7Effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on inflammatory factors in mice with DSS-induced inflammatory bowel disease. Detailed implementation manners

[0025] For the purpose of making the objectives and technical solutions of the implementation of the present invention clearer, the technical solutions in the embodiments of the present invention will be described in detail and clearly below. Those skilled in the art will understand that the following embodiments are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention.

[0026] The present invention screened out a small molecule compound - ethyl 4-hydroxy-3,5-dimethoxybenzoate (Table 1) with a significant inhibitory effect on intestinal smooth muscle contraction from 44 natural Chinese medicine small molecules (concentration 100 μM).

[0027] Table 1 Inhibitory effect of natural Chinese medicine small molecules on intestinal smooth muscle contraction Compound Inhibitory effect Compound Inhibitory effect Cinnamic acid - Methyl cinnamate - Ethyl cinnamate - Propyl cinnamate - Butyl cinnamate - Octyl cinnamate - Methyl ferulate - Ethyl ferulate + Propyl ferulate - Butyl ferulate - Isooctyl ferulate - Phloroglucinol - Hexyl ferulate - Eugenol - Gallic acid - Methyl gallate - Ethyl gallate + Methyl tanshinolate pivalate - Butyl gallate - Octyl gallate - Propyl gallate - Hexyl gallate - Caffeic acid - Methyl caffeate - Ethyl caffeate - Propyl caffeate - Butyl caffeate - Octyl caffeate - Isobutyl caffeate - Vanillic acid - Methyl vanillate - Ethyl vanillate - Propyl vanillate - Butyl vanillate - Isobutyl vanillate - Octyl vanillate - Methyl syringate - Syringic acid - Methyl tanshinolate - Tanshinol - Ethyl tanshinolate - Propyl tanshinolate - Ethyl 4-hydroxy-3,5-dimethoxybenzoate ++ Butyl tanshinolate - Note: When the inhibition rate is less than 30% at a concentration of 100 μM, it is marked as -; greater than 30% and less than 40% is marked as +; greater than 40% and less than 50% is marked as ++; greater than 50% is marked as +++.

[0028] The structural formula of ethyl 4-hydroxy-3,5-dimethoxybenzoate is as follows: .

[0029] Example 1: Therapeutic effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on diarrhea 1. Materials and reagents (1) Animals: KM mice, half male and half female, 18 - 22 g, purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd.; New Zealand rabbits, 2.0 - 2.5 kg, 75 days old, half male and half female, purchased from Qingdao Qinda Biotechnology Co., Ltd., all were raised in the animal laboratory of the School of Medicine, Yushan Campus, Ocean University of China.

[0030] (2) Formula of 1000 mL Tyrode's solution: NaCl 8.0 g, 10% KCl 2.0 ml (0.2 g), 10% MgSO 4 .7H 2 O 2.6 ml (0.26 g), 5% NaH 2 PO 4 ·2H 2 O 1.3 ml (0.065 g), NaHCO 3 1.0 g, 1M CaCl 2 1.8 ml (0.2 g), glucose 1.0 g (CaCl 2 was first dissolved separately with distilled water and then added to the solution).

[0031] (3) Preparation method of ethyl 4-hydroxy-3,5-dimethoxybenzoate: Suspension: Accurately weigh 150 mg of ethyl 4-hydroxy-3,5-dimethoxybenzoate (purchased), grind it, and suspend it in 0.5% CMC-Na to make the final concentrations 7.5 mg / mL and 15 mg / mL.

[0032] Injection: Accurately weigh 1500 mg of ethyl 4-hydroxy-3,5-dimethoxybenzoate, dissolve it in DMSO to prepare a stock solution of 750 mg / mL, and dilute it with normal saline to 2.5 mg / mL, 5.0 mg / mL, and 7.5 mg / mL.

[0033] (4) Preparation method of atropine sulfate (purchased from Aladdin Reagent): Accurately weigh 300 mg of atropine, dissolve it in DMSO to prepare a stock solution of 150 mg / mL, and dilute it with normal saline to 1.5 mg / mL when in use.

[0034] (5) Preparation method of loperamide hydrochloride (purchased from Aladdin Reagent): Accurately weigh 1 mg of loperamide hydrochloride, suspend it in 0.5% CMC-Na to make the final concentration 0.1 mg / mL.

[0035] (6) Decoction of senna leaves: Accurately weigh 50 g of senna leaves, add water to cover the senna leaves, heat to boil for 20 min, then filter and concentrate the obtained solution to 50 mL to obtain a 1 g / mL decoction of senna leaves.

[0036] (7) Preparation of 5% carbon ink: Accurately weigh 0.1 g of CMC-Na, dissolve it in 20 mL of double-distilled water, and add 1.053 mL of India ink after complete dissolution to obtain 5% carbon ink.

[0037] 2. Methods (1) Effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on spontaneous contraction of isolated rabbit duodenum To verify the antidiarrheal effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate, the effect of the drug on the activity of isolated rabbit intestinal smooth muscle was observed through in vitro experiments. The rabbits were fasted for 24 h without water deprivation in advance, killed by quickly hitting the head, dissected, the duodenum was taken out, placed in a petri dish containing Tyrode's solution, the surrounding mucosa was stripped, a 1.5 - 2 cm long small segment was cut, opened and connected to the BL-420E biological function test system, and Tyrode's solution and double-distilled water were added respectively, and preheated at 37°C. After calibration and zero adjustment, the tissue was placed under an initial load of 1.0 g, and air was passed, 2 - 3 per second. After equilibration for 20 - 30 min, the effect of the cumulative drug on the intestinal spontaneous contraction was observed.

[0038] (2)Effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on intestinal muscle contraction induced by acetylcholine chloride, histamine and barium chloride After the intestinal tube contraction was stable, acetylcholine chloride (final concentration 1 μmol / L, the following are all final concentrations), histamine (4 mg / mL) and barium chloride (12 mg / mL) were added respectively to induce intestinal muscle contraction, and then 100 μM ethyl 4-hydroxy-3,5-dimethoxybenzoate was added, and the amplitude and frequency of intestinal motility were observed and recorded.

[0039] (3)Effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on intestinal muscle contraction induced by calcium chloride The Tyrode's solution in the test tube was replaced with calcium-free high-potassium Tyrode's solution, and the intestinal tube was placed in it for 60 min to fully remove the calcium in the intestinal muscle, and then replaced with calcium-free Tyrode's solution. After equilibrium and stabilization, calcium chloride (500 μM) was added to induce intestinal muscle contraction, and then 100 μM ethyl 4-hydroxy-3,5-dimethoxybenzoate was added, and the average tension of intestinal motility was observed and recorded.

[0040] (4)Small intestine propulsion experiment To verify the inhibitory effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on the intestinal motility of normal mice, the small intestine propulsion test of animals was carried out. A total of 32 KM mice were divided into Control, loperamide (2 mg / kg), and ethyl 4-hydroxy-3,5-dimethoxybenzoate groups (150 mg / kg, 300 mg / kg). Before the experiment, the mice were fasted but allowed to drink water for 24 h. Each group was intragastrically administered the corresponding drug once, with a volume of 0.2 ml / 10 g. The control group was given an equal volume of normal saline. After 30 min of drug administration, 0.26 ml / animal of carbon ink was intragastrically administered. After 20 min, the mice were sacrificed, the abdominal cavity was quickly opened, and all intestinal tissues from the pylorus to the ileocecal junction were excised, the mesentery was cut off, flattened, and the total length and the advancing distance of the ink were measured.

[0041] The ink propulsion rate (%) = advancing distance of ink (cm) / total length of small intestine (cm) * 100%, and the ink propulsion rates of the small intestine in each dose group were obtained.

[0042] (5)Effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on senna-induced diarrhea in mice To verify the inhibitory effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on intestinal motility in diarrhea mice, senna was used to induce diarrhea in mice, and the small intestine propulsion test of animals was carried out. A total of 40 KM mice, half male and half female, were divided into a control group, a model group, an atropine group (30 mg / kg), and an ethyl 4-hydroxy-3,5-dimethoxybenzoate group (100 mg / kg). Except for the control group given normal saline, the other groups were intraperitoneally injected with the corresponding drug once, with a volume of 0.2 ml / 10 g. The control group was given an equal volume of normal saline. 0.5 ml of 1 g / mL senna water decoction was intragastrically administered 30 min after drug administration. 0.26 ml / animal of carbon ink was given 40 min later. After 20 min, the animals were sacrificed. The abdominal cavity was quickly opened, and all intestinal tissues from the pylorus to the ileocecal junction were excised. The mesentery was cut off, flattened, and the total length and the advancing distance of the ink were measured.

[0043] To verify the therapeutic effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on diarrhea mice, senna was used to induce diarrhea in mice. A total of 50 KM mice, half male and half female, were divided into a Control group, a model group, a loperamide (2 mg / kg) group, and an ethyl 4-hydroxy-3,5-dimethoxybenzoate group (150 mg / kg, 300 mg / kg). Except for the Control group given normal saline, the other groups were given 0.4 ml of 1 g / mL senna water decoction for 5 consecutive days. On the fifth day, 30 min after the senna water decoction was given, the corresponding drug was intragastrically administered once, with a volume of 0.2 ml / 10 g. The control group was given an equal volume of normal saline. They were placed in a 1 L beaker with filter paper placed underneath, and the number of loose stools, the total number of feces, and the loose stool grade of each group of mice were observed within 4 h. The loose stool grade was judged by the size of the stain formed by the feces on the filter paper.

[0044] (6)Data analysis Graphpad prism 8.0 software was used to statistically analyze the experimental data. The experimental results were expressed as mean ± SEM. One-way ANOVA was used to test the ink propulsion rate. There were significant differences between the blank group and the model group, and between the model group and each drug group.

[0045] 3. Results and analysis (1)Effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on the spontaneous contraction of isolated rabbit duodenum After the intestinal segment ran smoothly for 30 - 60 min, curves were obtained by the cumulative dosing method as Figure 1As shown in the figure, ethyl 4-hydroxy-3,5-dimethoxybenzoate (MD50) had little effect on the amplitude of intestinal smooth muscle movement at 10 μM. However, with each increase in concentration, the amplitude of intestinal smooth muscle movement gradually decreased, indicating that ethyl 4-hydroxy-3,5-dimethoxybenzoate could inhibit the spontaneous contraction of rabbit intestinal smooth muscle in a concentration-dependent manner, and inhibited 80% of the intestinal peristalsis function at 300 μM.

[0046] (2) Effects of ethyl 4-hydroxy-3,5-dimethoxybenzoate on intestinal muscle contractions induced by acetylcholine, histamine, barium chloride, and calcium chloride As can be seen from Figure 2, 100 μM could significantly inhibit the intestinal muscle contractions induced by acetylcholine, histamine, and barium chloride. At the same time, it could also directly inhibit the contraction induced by calcium chloride, indicating that the drug ethyl 4-hydroxy-3,5-dimethoxybenzoate played a role in relaxing smooth muscle by inhibiting the influx of extracellular calcium.

[0047] (3) Effects of ethyl 4-hydroxy-3,5-dimethoxybenzoate on small intestinal peristalsis in normal mice The results were as Figure 3A shown. 150 mg / kg of ethyl 4-hydroxy-3,5-dimethoxybenzoate could inhibit the intestinal peristalsis of mice, but there was a significant difference only at high doses.

[0048] (4) Effects of ethyl 4-hydroxy-3,5-dimethoxybenzoate on small intestinal peristalsis in diarrhea mice induced by senna leaf The results were as Figure 3B shown. 100 mg / kg of ethyl 4-hydroxy-3,5-dimethoxybenzoate could inhibit the intestinal peristalsis of diarrhea mice with a significant difference.

[0049] (5) Therapeutic effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on diarrhea mice induced by senna leaf The results are shown in Table 2. 300 mg / kg of ethyl 4-hydroxy-3,5-dimethoxybenzoate could significantly reduce the diarrhea index, showing a significant difference compared with the model group.

[0050] Table 2 Therapeutic effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on diarrhea mice induced by senna leaf

[0051] Example 2: Therapeutic effect of ethyl 4-hydroxy-3,5-dimethoxybenzoate on dextran sulfate sodium (DSS)-induced inflammatory bowel disease 1. Materials and reagents (1) Animals: C57BL / 6j mice, male, 18 - 22 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., and raised in the animal laboratory of the School of Medicine, Yushan Campus, Ocean University of China.

[0052] (2) 2% DSS (MP Biomedicals): Weigh 2 g of DSS and dissolve it in 100 mL of distilled water.

[0053] (3) Ethyl 4-hydroxy-3,5-dimethoxybenzoate: The preparation method is the same as that in Example 1.

[0054] (4) 500 mg / kg SASP (Shanghai Xinyi Tianping Pharmaceutical Co., Ltd.): Take 1 SASP tablet (250 mg) and grind it into powder. Add 150 μL of DMSO and 750 μL of HS15, and make up the volume to 15 mL with normal saline.

[0055] 2. Methods (1) Experimental procedure C57BL / 6j mice were divided into a control group (Con), a model group (Model), a positive drug group (500 mg / kg sulfasalazine), and a drug group of ethyl 4-hydroxy-3,5-dimethoxybenzoate (low-dose group 50 mg / kg, medium-dose group 100 mg / kg, high-dose group 150 mg / kg). After one week of adaptive feeding, the mice were allowed to freely drink 2% DSS solution for 7 consecutive days. At the same time, the Con and Model groups were given corresponding normal saline for 7 consecutive days, and the drug groups were given the corresponding doses of drugs. After 7 days, the mice were fasted for 12 h, blood was collected by eye socket puncture, and colon tissues were collected for subsequent experiments.

[0056] (2) Detection indicators ① Disease activity (DAI) index The body weight of the mice was recorded every day. DAI was determined by the degree of weight loss, stool consistency, and stool bleeding score. The DAI index is the sum of the three numbers of weight loss, stool consistency, and stool bleeding score. The scoring criteria are shown in Table 3.

[0057] Table 3 Clinical scoring criteria

[0058] ② Measure the colon length and weight, and the ratio of its weight to length can be used as an indirect indicator of inflammation.

[0059] ③ Spleen (liver) coefficient = spleen (liver) weight / mouse body weight.

[0060] ④ ELISA was used to detect the expression of related inflammatory factors in the serum of mice. The inflammatory factors include TNF-a and IL-6.

[0061] (3)Data analysis The experimental data were statistically analyzed using Graphpad prism 8.0 software. The experimental results were expressed as mean ± SEM, and one-way ANOVA was used to test the blind length, spleen index, liver index, and the expression of inflammatory factors in serum. There were significant differences between the blank group and the model group, and between the model group and each drug group for all indicators except the liver index, which showed no significant change.

[0062] 3. Results and analysis (1)Effect on the body weight of mice with inflammatory bowel disease As Figure 4 shown, during the entire experimental process, the body weight of the control group tended to be stable and showed a slow upward trend. The body weight of the model group began to decline after the third day and continued to decline. The body weight of the drug group also showed a downward trend, but the degree of decline was lower than that of the model group, indicating that ethyl 4-hydroxy-3,5-dimethoxybenzoate can improve the weight loss symptoms of mice with DSS-induced inflammatory bowel disease.

[0063] (2)Effect on the DAI index of mice with inflammatory bowel disease As Figure 5A shown, during the entire experimental process, there were no occurrences of bloody stools or loose stools in the control group. Starting from the third day of modeling, the model group began to show bloody stools and diarrhea, which gradually worsened, showing significant differences compared with the control group. Compared with the model group, the degrees of diarrhea and bloody stools in the ethyl 4-hydroxy-3,5-dimethoxybenzoate and SASP groups were significantly reduced, indicating that both ethyl 4-hydroxy-3,5-dimethoxybenzoate and SASP can significantly improve the diarrhea and bloody stool symptoms of DSS-induced inflammatory bowel disease.

[0064] (3)Effect on the colon length of mice with inflammatory bowel disease As Figure 5B shown, compared with the control group, the colon of mice in the model group showed swelling, congestion, and was significantly shortened. The colon lengths of ethyl 4-hydroxy-3,5-dimethoxybenzoate and SASP were significantly longer than those of the model group, indicating that ethyl 4-hydroxy-3,5-dimethoxybenzoate and SASP can significantly improve the colon shortening symptoms of mice with DSS-induced inflammatory bowel disease.

[0065] (4)Effect on the organ coefficients of mice with inflammatory bowel disease As Figure 6 shown, compared with the control group, the spleen coefficient of mice in the model group was significantly increased, and the coefficient decreased correspondingly after drug treatment. Moreover, there was no significant effect on the liver coefficient after drug treatment, indicating that ethyl 4-hydroxy-3,5-dimethoxybenzoate and SASP can significantly reduce the spleen coefficient of mice with DSS-induced inflammatory bowel disease.

[0066] (6) Effects on inflammatory factors in the blood of mice with inflammatory bowel disease As Figure 7 shown, compared with the control group, the levels of TNF-a and IL-6 in the serum of the model group were significantly increased, while those in the 4-hydroxy-3,5-dimethoxybenzoyl ethyl ester and SASP groups were significantly lower than those in the model group, indicating that 4-hydroxy-3,5-dimethoxybenzoyl ethyl ester and SASP can significantly improve the inflammatory response in DSS-induced inflammatory bowel disease mice.

[0067] In summary, through in vivo and in vitro experiments, the present invention confirms that 4-hydroxy-3,5-dimethoxybenzoyl ethyl ester has a good therapeutic effect on senna-induced diarrhea in mice, can effectively inhibit intestinal peristalsis, improve diarrhea symptoms, and also inhibit intestinal peristalsis in normal mice. It is a safe and effective antidiarrheal drug. At the same time, 4-hydroxy-3,5-dimethoxybenzoyl ethyl ester also has a good therapeutic effect on DSS-induced inflammatory bowel disease, can effectively reduce the incidence of diarrhea and bloody stools, and inhibit the production of inflammatory factors. It is an effective drug for treating IBD. The present invention reports for the first time the effects of 4-hydroxy-3,5-dimethoxybenzoyl ethyl ester on diarrhea and inflammatory bowel disease, providing a new candidate compound for the development of antidiarrheal and inflammatory bowel disease drugs.

[0068] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, for those of ordinary skill in the art, it is still possible to modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions required to be protected by the present invention.

Claims

1. Use of a natural benzoic acid compound in the preparation of a medicament for treating gastrointestinal diseases.

2. The use according to claim 1, wherein, the natural benzoic acid compound is ethyl 4-hydroxy-3,5-dimethoxybenzoate, and its structural formula is: 。 3. The use according to claim 2, wherein, the gastrointestinal diseases include diarrhea and inflammatory bowel disease.

4. The use according to claim 2, wherein, the dosage of ethyl 4-hydroxy-3,5-dimethoxybenzoate is 10 μM - 300 μM.

5. The use according to claim 2, wherein, ethyl 4-hydroxy-3,5-dimethoxybenzoate can inhibit the spontaneous contraction of intestinal smooth muscle, inhibit intestinal peristalsis, and reduce the diarrhea index, so as to achieve the effect of treating diarrhea.

6. The use according to claim 2, wherein, ethyl 4-hydroxy-3,5-dimethoxybenzoate plays a role in relaxing smooth muscle by inhibiting the influx of extracellular calcium.

7. The use according to claim 2, wherein, ethyl 4-hydroxy-3,5-dimethoxybenzoate can improve the diarrhea, bloody stool symptoms, and colon shortening symptoms of inflammatory bowel disease, reduce the spleen coefficient of patients with inflammatory bowel disease, and the expression of inflammatory factors in the blood.

8. A medicament for treating gastrointestinal system diseases, wherein, the active ingredient in the medicament is compound ethyl 4-hydroxy-3,5-dimethoxybenzoate or its stereoisomer, pharmaceutically acceptable salt.

9. The medicament according to claim 8, wherein, the medicament is a tablet, capsule, dripping pill, granule, injection, powder injection or aerosol.

10. The medicament according to claim 8, wherein, the administration methods of the medicament include oral administration, intramuscular injection, intravenous injection or infusion, transdermal administration, mucosal administration, aerosol administration.

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