Application of 4-cholestene-3-one in preparing medicine for preventing or treating inflammatory bowel disease

Abstract

The present invention discloses the use of 4-cholesten-3-one in the preparation of drugs for preventing or treating inflammatory bowel disease. By establishing a mouse model of inflammatory bowel disease (IBD) induced by dextran sulfate sodium (DSS), the effects of 4-cholesten-3-one on the weight change, stool viscosity and stool bleeding of mice, the calculated disease activity index (DAI), and the length of the colon were observed. The results showed that 4-cholesten-3-one had a significant therapeutic effect on DSS-induced colitis, providing a new drug for the clinical treatment of IBD.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to application of 4-cholestene-3-one in preparing a drug for preventing or treating inflammatory bowel disease. Background Art

[0002] Inflammatory Bowel Disease (IBD) is a chronic nonspecific intestinal inflammatory disease mainly caused by disorders of the intestinal microbiota and immune system, including Crohn's disease (CD) and ulcerative colitis (UC). Currently, the treatment of IBD mainly relies on traditional therapeutic drugs: including 5-aminosalicylic acid, glucocorticoids, immunomodulators, etc. Traditional drugs have poor control effects on some moderate to severe patients and have serious side effects. Therefore, finding an active and effective treatment for IBD is a clinical problem that needs to be urgently addressed.

[0003] As the second largest class of drugs after antibiotics, steroid compounds occupy an important position in medicine and are widely used to treat cardiovascular diseases, anti-tumor, anti-infection, anti-allergy, etc. 4-cholestene-3-one is a precursor substance for a series of steroid hormone drugs, a metabolic intermediate of intestinal bacteria, and participates in lipid metabolism. It can also be used directly as a drug, has certain health functions and potential medicinal value, and can be used to inhibit the accumulation of fat in the human body, treat liver disease, and prevent skin keratinization. However, there is no literature report on the use of 4-cholestene-3-one in the preparation of drugs for the prevention or treatment of inflammatory bowel disease. Summary of the invention

[0004] In view of this, the purpose of the present invention is to provide the use of 4-cholesten-3-one in the preparation of drugs for preventing or treating inflammatory bowel disease, so as to provide new drugs for the clinical treatment of inflammatory bowel disease.

[0005] The inflammatory bowel disease (IBD) described in the present invention includes ulcerative colitis and Crohn's disease.

[0006] The 4-cholesten-3-one of the present invention can be used in combination with other drugs for preventing and treating inflammatory bowel disease, and can also be used as the only active ingredient.

[0007] The present invention induces an IBD mouse model by using dextran sulfate sodium (DSS), detects changes in mouse weight, stool viscosity and stool bleeding, calculates the disease activity index (DAI), and observes the effect of 4-cholesten-3-one (4CTL3) on the length of the mouse colon. The results show that 4-cholesten-3-one has no obvious reversal effect on the weight loss caused by colitis. By observing the stool viscosity and stool bleeding of mice every day, the DAI is calculated, and it is found that the DAI score of the model group is significantly higher than that of the 4CTL3 group; the length of the mouse colon is measured, and it is found that 4CTL3 can significantly prolong the colon length of IBD mice. The above results show that 4-cholesten-3-one (4CTL3) has a significant therapeutic effect on DSS-induced colitis and can significantly improve the symptoms of IBD.

[0008] The drug of the present invention comprises an effective amount of 4-cholesten-3-one and a pharmaceutically acceptable carrier. The drug for preventing or treating inflammatory bowel disease of the present invention can be prepared into a suitable dosage form by conventional methods in the art. The pharmaceutically acceptable carrier is selected from pharmaceutically acceptable suspensions. Thus, the drug can be prepared and formed into a form suitable for administration of a clinical pharmaceutical preparation.

[0009] Preferably, the dosage form of the drug is a suspension.

[0010] Compared with the prior art, the present invention has the following excellent effects:

[0011] The present invention provides a new use of 4-cholesten-3-one in the preparation of a drug for preventing or treating inflammatory bowel disease. By establishing an IBD mouse model induced by dextran sulfate sodium (DSS), the results show that 4-cholesten-3-one has a significant therapeutic effect on DSS-induced colitis and can significantly improve IBD symptoms, thereby providing a new drug for the clinical treatment of IBD. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 The effect of 4CTL3 on the body weight of colitis mice. *** P<0.001 (model group vs blank group); ### P<0.001 (4CTL3 group vs model group).

[0013] Figure 2 The effect of 4CTL3 on the disease activity index of colitis mice. *** P<0.001 (model group vs blank group); ## P<0.01 (4CTL3 group vs model group).

[0014] Figure 3 The effect of 4CTL3 on colon length in colitis mice. *** P<0.001 (model group vs blank group); ## P<0.01, ### P<0.001 (5-ASA, 4CTL3 group vs model group). DETAILED DESCRIPTION

[0015] The present invention is further described below by means of specific implementation methods. The following examples are specific implementation methods of the present invention, but the implementation methods of the present invention are not limited to the following examples.

[0016] Example 1: Therapeutic effect of 4-cholesten-3-one on IBD in mice

[0017] 1. Materials and Methods

[0018] 1.1 Drugs

[0019] (+)-4-cholesten-3-one (4CTL3) was purchased from Sigma (CAS: 601-57-0) and prepared into a suspension using 5% CMCNA; sulfasalazine (lot: J2212276) was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. Dextran sulfate sodium was purchased from MP Biomedicals (lot: S7980).

[0020] 1.2 Instruments

[0021] MTB1000D electronic balance (Shenzhen Meifu Electronics Co., Ltd.)

[0022] 1.3 Animals

[0023] C57BL / 6 male mice (purchased from Guangdong Medical Experimental Animal Center, license number SCXK (Yue) 2022-0002 Yuejianzhengzi 2006A018), weighing 19-21 g. Mice were housed in the Experimental Animal Center of Guangdong Medical University under 12 h light / darkness and free access to food.

[0024] 1.4 Methods

[0025] After 7 days of adaptive feeding with standard diet and water (24-25°C, humidity 70-75%, 12 hours of light / darkness), the mice were randomly divided into 4 groups, 6 in each group, including control group, model group, 5-ASA group (200 mg / kg), and 4CTL3 group (50 mg / kg). The 4CTL3 group was given preventive oral administration for 5 days, the model group was given oral purified water for 5 consecutive days (from day 0 to day 5), and on day 5, the mice drank 2.5% DSS freely and continued to be administered for 7 days (from day 5 to day 12). The blank group drank purified water freely. The weight, stool viscosity and fecal bleeding of the mice were observed and recorded every day. On day 12, the anesthetized mice were killed after blood was taken. The length of the mouse colon tissue was measured.

[0026] Evaluation by disease activity index (DAI): The disease activity index score was determined according to the weight loss (no loss was recorded as 0 points, loss of 1% to 5% was recorded as 1 point, loss of 5% to 10% was recorded as 2 points, loss of 10% to 5% was recorded as 3 points, and loss greater than 15% was recorded as 4 points), stool viscosity (normal stool was recorded as 0 points, soft stool was recorded as 1 point, loose and wet stool was recorded as 2 points, diarrhea and dry stool was recorded as 3 points, diarrhea and wet stool was recorded as 4 points), and fecal bleeding of IBD mice (no blood was recorded as 0 points, slight blood was recorded as 1 point, occult blood was recorded as 2 points, little bleeding was recorded as 3 points, and heavy bleeding was recorded as 4 points). DAI calculation: DAI = (weight loss score + stool viscosity score + stool blood degree score) / 3

[0027] 1.5 Statistical methods

[0028] The results were analyzed using Graphpad Prism 7.0 statistical analysis software. The data of each group are expressed as x-±SD, and the t-test was used for comparison between samples.

[0029] 2. Results

[0030] 2.1 Effect of 4CTL3 on the body weight of IBD mice

[0031] The experimental results showed that 4CTL3 had no significant effect on the body weight caused by IBD. There was a significant difference in body weight between the 4CTL3 group and the model group (P<0.001), and there was a significant difference in body weight between the model group and the blank group (P<0.001). The results are shown in Table 1. Figure 1 .

[0032] Table 1.4 Effect of CTL3 on body weight of colitis mice (x-±SD N=6)

[0033]

[0034] *** P<0.001 (model group vs blank group); ### P<0.001 (4CTL3 group vs model group).

[0035] 2.2 Effect of 4CTL3 on the disease activity index of colitis mice

[0036] After 4CTL3 treatment, the symptoms of diarrhea, rectal bleeding and weight loss in IBD mice were significantly alleviated, and the DAI score was significantly reduced. There was a significant difference in DAI between the 4CTL3 group and the model group (P<0.01). The DAI value of the 4CTL3 group was significantly lower than that of the model group. There was a significant difference in DAI between the model group and the blank group (p<0.010). The results are shown in Table 2. Figure 2 .

[0037] Table 2.4 Effect of CTL3 on the disease activity index of colitis mice (x-±SD N=6)

[0038]

[0039] *** p<0.001 (model group vs blank group); ## P<0.01 (4CTL3 group vs model group).

[0040] 2.3 Effect of 4CTL3 treatment on colon length

[0041] Compared with the blank group, the colon length of the model group was significantly shortened (P<0.001), and after 12 days of 4CTL3 treatment, the shortening of colon length was significantly reversed, and there was a significant difference in colon length between the 4CTL3 group and the model group (p<0.001). The results are shown in Table 3. Figure 3 .

[0042] Table 3.4 Effect of CTL3 on colon length in colitis mice (x-±SD N=6)

[0043]

[0044] *** p<0.001 (model group VS blank group); ## P<0.01, ### P<0.001 (5-ASA, 4CTL3 group vs model group).

[0045] In summary, by establishing an IBD mouse model induced by sodium dextran sulfate, the 4CTL3 group showed significant differences compared with the model group in terms of DAI score and colon length. It can be concluded that 4CTL3 has a significant therapeutic effect on DSS-induced colitis, can significantly improve IBD symptoms, and provides a new drug for the clinical treatment of IBD.

Claims

1. Use of 4-cholestene-3-one in the preparation of drugs for preventing or treating inflammatory bowel disease; the inflammatory bowel disease is ulcerative colitis and Crohn's disease.

2. The use according to claim 1, characterized in that: The 4-cholestene-3-one is used in combination with other drugs for preventing and treating inflammatory bowel disease.

3. The use according to claim 1, characterized in that: The 4-cholesten-3-one is used as the sole active ingredient.

4. The use according to claim 1, characterized in that: The medicine contains effective content of 4-cholesten-3-one and a pharmaceutically acceptable carrier.

5. The use according to claim 1, characterized in that: The dosage form of the drug is a suspension.

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