Preparation method and application of medicine for treating inflammatory bowel disease

Abstract

The invention discloses a preparation method and application of a medicine for treating inflammatory bowel disease, and relates to the technical field of medical treatment, the medicine comprises guaijave glycoside, dimethyl sulfoxide and sterile corn oil, and the medicine comprises the following components: 20mg of guaijave glycoside, 200mu l of dimethyl sulfoxide and 1800mu l of sterile corn oil. The guaijave glycoside can improve the general conditions (weight loss and bloody stool) and the severity of intestinal injury of a mouse with the inflammatory bowel disease induced by DSS, and effectively relieve the intestinal inflammation of the mouse with the inflammatory bowel disease.

Description

Technical field:

[0001] The present invention relates to the field of medical technology, and in particular to a medicine for treating inflammatory bowel disease, a preparation method thereof and an application thereof. Background technology:

[0002] In recent years, the number of patients with inflammatory bowel disease (IBD) has been increasing worldwide and will reach its peak by 2030, seriously endangering the health of patients and becoming a global public health issue. Currently, there are a variety of drugs for the treatment of IBD, but they all have many problems such as poor safety, poor effectiveness, and high prices. Developing a new, safe, effective, and inexpensive drug is the focus and difficulty of this research field.

[0003] Guava belongs to the Myrtaceae family and is mainly distributed in Hainan, Guangdong, Guangxi, Yunnan and other regions in my country. The roots, meridians, leaves and fruits of guava have medicinal value. The fruit of guava is rich in antioxidants, vitamin C, potassium and dietary fiber. In my country, India, Pakistan and Bangladesh, the main use of guava leaf decoction is to treat diarrhea, diabetes, cardiovascular disease, tumors, inflammatory diseases, cough and sputum and parasitic infection. At present, the research field of food and medicine mainly studies the chemical composition and pharmacological effects of guava extract complex. The complex contains polyphenols, flavonoids, terpenes and glycoside compounds, which have antibacterial, anti-infective, anti-inflammatory, detumescent and anti-tumor effects. Guava glycosides are Chinese medicine monomers extracted from guava complexes. In the dextran sulfate sodium salt (DSS)-induced mouse IBD animal model, oral administration of guava glycosides (Gua) has the effects of improving mouse weight loss, protecting mice from intestinal damage and inhibiting the expression of intestinal proinflammatory factors. Guava glycosides are expected to be used clinically as a new drug for the treatment of IBD.

[0004] The disadvantages of the prior art are that 5-aminosalicylic acid drugs can cause side effects such as nausea, vomiting, headache, reversible male infertility, rash, granulocytopenia, autoimmune hemolysis, aplastic anemia, etc.; glucocorticoids can cause side effects such as osteoporosis; and immunosuppressants can suppress the patient's immune function, leading to increased susceptibility of the patient. In order to solve the above technical problems, a new technical solution is proposed. Summary of the invention:

[0005] The purpose of the present invention is to overcome the deficiencies of the above-mentioned prior art, and to propose a preparation method and application of a drug for treating inflammatory bowel disease. The main purpose of clinical treatment of inflammatory bowel disease is to control acute attacks, promote mucosal healing, maintain remission, reduce recurrence, and prevent and treat complications. The main drugs for clinical treatment of inflammatory bowel disease are 5-aminosalicylic acid, glucocorticoids and immunosuppressants. The above-mentioned drugs have many adverse drug reactions, strong side effects or expensive drug prices.

[0006] The technical solution adopted by the present invention is: a drug for treating inflammatory bowel disease, characterized in that: the drug includes guava glycosides, dimethyl sulfoxide, and sterile corn oil. The drug includes guava glycosides, dimethyl sulfoxide and sterile corn oil. The content of each component of the drug is 20 mg of guava glycosides, 200 ul of dimethyl sulfoxide, and 1800 ul of sterile corn oil.

[0007] As a preferred technical solution, the guava glycoside is quercetin-3-O-α-L-arabinoside, which is a traditional Chinese medicine monomer extracted from pomegranate peel. It is a yellow powder, slightly soluble in methanol, and easily soluble in dimethyl sulfoxide. The molecular formula is: C20H18O11, and the molecular weight is: 434.35.

[0008] A method for preparing a drug for treating inflammatory bowel disease, the method comprising the following steps:

[0009] Step 1, preparing guava glycoside drug solution;

[0010] Step 2, preparing a control drug solvent;

[0011] Step 3: Prepare drugs for inducing inflammatory bowel disease model.

[0012] As a preferred technical solution, the guava glycoside drug solution is prepared by dissolving 20 mg of guava glycoside in 200 ul of dimethyl sulfoxide, and then adding 1800 ul of corn oil at a ratio of 1:9, and vortexing to mix to make a drug solution with a concentration of 10 mg / ml.

[0013] As a preferred technical solution, the control drug solvent is prepared by taking 200ul of dimethyl sulfoxide, adding 1800ul of corn oil, and vortexing to mix, so as to prepare a drug solvent with a ratio of dimethyl sulfoxide to corn oil of 1:9.

[0014] As a preferred technical solution, the drug for inducing an inflammatory bowel disease mouse model is prepared by weighing 5 g of DSS and dissolving it in 100 ml of pure water to prepare a 5% DSS solution.

[0015] A drug application for treating inflammatory bowel disease, characterized in that: 12 mice are taken and divided into a Con group, a Gua group, a DSS group and a Gua+DSS group, with 3 mice in each group. Two days before the DSS-induced inflammatory bowel disease mouse model is established, the mice are weighed and administered once, and the Con group and the DSS group are given 100ul of a prepared control drug solvent by oral gavage. The Gua group and the Gua+DSS group are given a prepared guava glycoside (Gua) drug solution at 50 mg / kg by oral gavage. DSS-induced inflammatory bowel disease mouse model: The drinking water of mice in the DSS group and Gua+DSS group was replaced with 5% DSS solution; the drinking water of mice in the Con group and Gua group was replaced with pure water for 1 week. On days 0, 3, and 5 of the DSS-induced inflammatory bowel disease mouse model, the Con group and DSS group were given 100ul of the prepared control drug solvent by gavage, and the Gua group and Gua+DSS group were given 50mg / kg of guava glycoside (Gua) drug solution by gavage. On days 0, 2, 4, and 7 after the DSS-induced inflammatory bowel disease mouse model was established, the weight of each mouse was weighed and recorded. The weight changes of the mice and the presence of bloody stools were observed.

[0016] The beneficial effects of the present invention are as follows: Pharmacological experiments have shown that guava glycosides can improve the general condition (weight loss, bloody stools) and severity of intestinal damage in DSS-induced inflammatory bowel disease mice, and effectively alleviate intestinal inflammation in inflammatory bowel disease mice. Description of the drawings:

[0017] Figure 1 The weight changes and bloody stools of the mice of the present invention;

[0018] Figure 2 Schematic diagram of colon length;

[0019] Figure 3 This is a schematic diagram of the intestinal pathological changes observed by HE chromosomes of the present invention;

[0020] Figure 4 Schematic diagram of the mRNA expression level of inflammatory factors in the mouse colon of the present invention; Specific implementation method:

[0021] The present invention is further described with reference to specific embodiments:

[0022] A drug for treating inflammatory bowel disease, characterized in that the drug comprises guava glycosides, dimethyl sulfoxide, and sterile corn oil. The drug comprises guava glycosides, dimethyl sulfoxide, and sterile corn oil. The content of each component of the drug is 20 mg of guava glycosides, 200 ul of dimethyl sulfoxide, and 1800 ul of sterile corn oil.

[0023] As a preferred technical solution, the guava glycoside is quercetin-3-O-α-L-arabinoside, which is a traditional Chinese medicine monomer extracted from pomegranate peel. It is a yellow powder, slightly soluble in methanol, and easily soluble in dimethyl sulfoxide. The molecular formula is: C20H18O11, and the molecular weight is: 434.35.

[0024] A method for preparing a drug for treating inflammatory bowel disease, the method comprising the following steps:

[0025] Step 1, preparing guava glycoside drug solution;

[0026] Step 2, preparing a control drug solvent;

[0027] Step 3: Prepare drugs for inducing inflammatory bowel disease model.

[0028] As a preferred technical solution, the guava glycoside drug solution is prepared by dissolving 20 mg of guava glycoside in 200 ul of dimethyl sulfoxide, and then adding 1800 ul of corn oil at a ratio of 1:9, and vortexing to mix to make a drug solution with a concentration of 10 mg / ml.

[0029] As a preferred technical solution, the solution is prepared by weighing 5 g of sodium dextran sulfate and dissolving it in 100 ml of pure water to prepare a 5% sodium dextran sulfate solution.

[0030] An application of a drug for treating inflammatory bowel disease, the specific application includes the following steps: preparing the drug, grouping, weighing, administering, observing the situation, evaluating the degree of intestinal damage, statistical analysis and finally drawing conclusions. The specific steps are analyzed in detail below:

[0031] 1. Prepare drugs: Prepare 5% DSS solution: Weigh 5g DSS and dissolve it in 100ml pure water to prepare 5% DSS solution; wherein DSS is sodium dextran sulfate. Prepare guava glycoside (Gua) drug solution: Dissolve 20mg guava glycoside in 200ul DMSO, then add 1800ul corn oil (1:9), vortex mix to make a drug solution with a concentration of 10mg / ml. The ratio of DMSO to corn oil in the drug solvent is 1:9. Prepare control drug solvent: Take 200ul DMSO, add 1800ul corn oil, vortex mix, and prepare a drug solvent with a ratio of DMSO to corn oil of 1:9; wherein DMSO is dimethyl sulfoxide.

[0032] 2. Grouping: 12 mice were divided into Con group, Gua group, DSS group and Gua+DSS group, with 3 mice in each group;

[0033] 3. Weighing: Weigh and record the weight of each mouse on days 0, 2, 4, and 7 after DSS-induced inflammatory bowel disease mouse modeling;

[0034] 4. Administration: 2 days before the establishment of the DSS-induced inflammatory bowel disease mouse model, the mice were weighed and administered once. The Con group and the DSS group were given 100ul of the prepared control drug solvent by gavage. The Gua group and the Gua+DSS group were given 50mg / kg of the prepared guava glycoside (Gua) drug solution by gavage. To establish the DSS-induced inflammatory bowel disease mouse model: the drinking water of the mice in the DSS group and the Gua+DSS group was replaced with a 5% DSS solution; the drinking water of the mice in the Con group and the Gua group was replaced with pure water for 1 week. On days 0, 3, and 5 of the DSS-induced inflammatory bowel disease mouse model, the Con group and the DSS group were given 100ul of the prepared control drug solvent by gavage, and the Gua group and the Gua+DSS group were given 50mg / kg of the prepared guava glycoside (Gua) drug solution by gavage. On days 0, 2, 4, and 7 after the establishment of the DSS-induced inflammatory bowel disease mouse model, the weight of each mouse was weighed and recorded. The weight changes and bloody stools of the mice were observed.

[0035] 5. Observe the condition of mice: weight changes and bloody stools of mice, refer to Figure 1 ;

[0036] 6. Evaluation of intestinal damage: The mice were killed by dislocation of the neck on the 7th day after modeling, and the length of the colon was measured; refer to Figure 2 The colon of mice, about 0.5 cm long, was taken from 1.0 cm near the cecum and fixed with formaldehyde for HE staining of pathological sections; Figure 3 The colon of mice, about 1.0 cm long, 1.5 cm near the cecum, was taken and the mRNA expression level of inflammatory factors was detected by qPCR; Figure 4 .

[0037] 7. Analysis: The experimental data and results were analyzed using Prism statistical software. The data were expressed as mean ± standard error of the mean. One-way analysis of variance was used. P < 0.05 indicated that the difference was statistically significant. P < 0.05; P < 0.01; P < 0.001; NS showed no statistical difference.

[0038] 8. Conclusion: Compared with the DSS group, the DSS+Gua group improved the weight loss of mice. Figure 1 A; Compared with the DSS group, the DSS+Gua group improved the bloody stool of mice. Figure 1 B. Compared with the DSS+Gua group, the colon length of mice in the DSS group was shortened, thickened, and congested. Figure 2 HE staining was used to observe the pathological changes of intestinal tissues in each group. Compared with the Con group, the DSS group showed a large number of inflammatory cell infiltration, mucosal epithelial cells and tissue necrosis and other pathological changes, while the inflammatory cell infiltration, mucosal epithelial cells and tissue necrosis in the DSS+Gua group were improved; Figure 3. The expression levels of inflammatory factors in the colon of mice were evaluated. Compared with the Con group, the mRNA expression of inflammatory factors IL-6, IFN-γ, TNF-α and COX-2 in the DSS group was upregulated. The DSS+Gua group significantly inhibited the mRNA expression levels of IL-6, IFN-γ, TNF-α and COX-2. Figure 4 .

[0039] The above results indicate that Gua improved intestinal damage in DSS-induced ulcerative colitis mice, and guava glycosides can improve the general condition (weight loss and bloody stools) and severity of intestinal damage in the DSS-induced inflammatory bowel disease mouse model, and effectively alleviate intestinal inflammation in inflammatory bowel disease mice.

[0040] For those skilled in the art, various other corresponding changes and modifications can be made according to the above technical solutions and concepts, and all of these changes and modifications should fall within the protection scope of the claims of the present invention.

Claims

1. A drug for treating inflammatory bowel disease, characterized in that: The drug comprises guava glycoside, dimethyl sulfoxide and sterile corn oil, and the content of each component of the drug is 20 mg of guava glycoside, 200 ul of dimethyl sulfoxide and 1800 ul of sterile corn oil.

2. The drug for treating inflammatory bowel disease according to claim 1, characterized in that: The guava glycoside is quercetin-3-O-α-L-arabinoside, which is a Chinese medicine monomer extracted from pomegranate peel. It is a yellow powder, slightly soluble in methanol, and easily soluble in dimethyl sulfoxide. The molecular formula is: 20 H 18 O 11 , molecular weight: 434.

35.

3. A drug for inducing an inflammatory bowel disease mouse model and a method for preparing a drug for treating inflammatory bowel disease, characterized in that: The method comprises the following steps: Step 1, preparing guava glycoside drug solution; Step 2, preparing a control drug solvent; Step 3: Prepare drugs for inducing inflammatory bowel disease model.

4. The method for preparing a drug for treating inflammatory bowel disease according to claim 3, characterized in that: The guava glycoside drug solution is prepared by dissolving 20 mg of guava glycoside in 200 ul of dimethyl sulfoxide, and then adding 1800 ul of corn oil at a ratio of 1:9, and vortexing to mix to make a drug solution with a concentration of 10 mg / ml.

5. The method for preparing a drug for treating inflammatory bowel disease according to claim 3, characterized in that: The control drug solvent was prepared by adding 1800 ul corn oil to 200 ul of dimethyl sulfoxide and vortex mixing to prepare a drug solvent with a ratio of dimethyl sulfoxide to corn oil of 1:

9.

6. The method for preparing the drug for inducing an inflammatory bowel disease mouse model according to claim 3: weigh 5 g of DSS, dissolve it in 100 ml of pure water, and prepare a 5% DSS solution.

7. A use of a drug for treating inflammatory bowel disease, characterized in that: Twelve mice were taken and divided into Con group, Gua group, DSS group and Gua+DSS group, with 3 mice in each group. Two days before the establishment of DSS-induced inflammatory bowel disease mouse model, the mice were weighed and administered once. The Con group and DSS group were given 100ul of the prepared control drug solvent by gavage, and the Gua group and Gua+DSS group were given 50mg / kg of the prepared guava glycoside (Gua) drug solution by gavage. To establish the DSS-induced inflammatory bowel disease mouse model: the drinking water of the mice in the DSS group and Gua+DSS group was replaced with a 5% DSS solution; the drinking water of the mice in the Con group and Gua group was replaced with pure water for 1 week. On days 0, 3, and 5 of the DSS-induced inflammatory bowel disease mouse model, the Con group and DSS group were given 100ul of the prepared control drug solvent by gavage, and the Gua group and Gua+DSS group were given 50mg / kg of the prepared guava glycoside (Gua) drug solution by gavage. On days 0, 2, 4, and 7 after DSS-induced inflammatory bowel disease mouse modeling, the body weight of each mouse was weighed and recorded. The weight changes and bloody stools of the mice were observed.

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