WO2024078507A1

WO2024078507A1 - Use of ergosterol in preparation of drug for preventing and treating gastric ulcers

Info

Publication number: WO2024078507A1
Application number: PCT/CN2023/123828
Authority: WO
Inventors: 许艺飞; 郭绍举; 黄萍; 洪欣欣; 李海文; 李娟娟
Original assignee: 深圳市中医院
Priority date: 2022-10-10
Filing date: 2023-10-10
Publication date: 2024-04-18
Also published as: CN115531393A

Abstract

The present invention belongs to the technical field of biological medicines, and specifically relates to the use of ergosterol in the preparation of a drug for preventing and treating gastric ulcers. The ergosterol of the present invention can promote the secretion of gastric mucus, resist gastric ulcers caused by absolute ethyl alcohol, promote the rehabilitation of gastric ulcers caused by acetic acid, and effectively prevent and treat gastric ulcers.

Description

Application of ergosterol in the preparation of drugs for preventing and treating gastric ulcer

This application claims the priority of the Chinese patent application filed with the China Patent Office on October 10, 2022, with application number CN202211239995.8 and invention name "Application of ergosterol in the preparation of drugs for the prevention and treatment of gastric ulcers", the entire contents of which are incorporated by reference in this application.

Technical Field

The invention belongs to the technical field of biomedicine, and particularly relates to the application of ergosterol in the preparation of medicines for preventing and treating gastric ulcers.

Background technique

Gastric ulcer is a common clinical disease. There are many factors that cause gastric ulcer, including drugs and diet (excessive drinking, use of nonsteroidal anti-inflammatory drugs, etc.), mental factors (high life pressure, high mental tension, etc.), genetic factors, etc. If gastric ulcer is not treated in time, it may also induce upper gastrointestinal bleeding, gastric perforation, pyloric obstruction and even gastric cancer. Clinically, proton pump inhibitors are often used to treat gastric ulcers. The treatment time is usually 1 to 2 months. Long-term use of proton pump inhibitors will inhibit the secretion of pepsin and also have a certain negative impact on the acid-secreting glands. Accelerating the recovery of gastric ulcers and reducing the use of proton pump inhibitors are one of the important clinical problems to be solved.

Ergosterol, also known as ergosterol, is a white or colorless shiny small leaf crystal or white crystalline powder. As an important component of fungal cell membrane, ergosterol has a stable structure and strong specificity. It is more representative than glucosamine for measuring biomass. Due to its unique physiological effects, ergosterol is widely used in drug development, but there is no record of ergosterol in the treatment of gastric ulcers.

Summary of the invention

The purpose of the present invention is to enrich the medical uses of ergosterol and effectively prevent and treat gastric ulcer.

In order to achieve the above purpose, the present invention provides the use of ergosterol in preparing a drug for preventing and treating gastric ulcer.

Preferably, the ergosterol is the only active ingredient.

Preferably, the triggering factors of gastric ulcer include alcohol and/or acid.

Preferably, the alcohol is anhydrous ethanol.

Preferably, the acid is acetic acid.

Preferably, the target of the drug is Rorα protein.

Preferably, the prevention and treatment of gastric ulcer includes promoting gastric mucus secretion and/or activating gastric stem cells.

Preferably, the dosage form of the drug includes any one of powder, injection, capsule, tablet, pill, granule and oral solution.

The present invention also provides a medicine for preventing and treating gastric ulcer, which comprises ergosterol and medically acceptable auxiliary materials.

Preferably, the ergosterol is used as the only active ingredient of the drug.

The present invention also provides a method for preventing and treating gastric ulcer, comprising administering ergosterol by intragastric administration at a dosage of 50-500 mg ergosterol/d/person.

The present invention provides the use of ergosterol in the preparation of a drug for preventing and treating gastric ulcers. The ergosterol of the present invention can promote the secretion of gastric mucus, resist gastric ulcers caused by anhydrous ethanol, and promote the recovery of gastric ulcers caused by acetic acid, and can be used for preventing and treating gastric ulcers.

BRIEF DESCRIPTION OF THE DRAWINGS

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required to be used in the embodiments are briefly introduced below.

FIG1 shows the preventive effect of ergosterol on acute gastric ulcer in rats induced by anhydrous ethanol;

FIG2 shows the therapeutic effect of ergosterol on gastric ulcer in mice induced by acetic acid;

FIG3 is the molecular docking result of ergosterol and RoRα on gastric stem cells;

FIG4 shows the change of the root mean square deviation (RMSD) of the complex over time during the molecular dynamics simulation.

Detailed ways

The invention provides application of ergosterol in preparing medicine for preventing and treating gastric ulcer.

The ergosterol described in the present invention is a type of component with cyclopentane polyhydrophenanthrene as the parent nucleus, and has a similar skeleton to corticosteroids. The specific structural formula is as follows:

In the present invention, the purity of the ergosterol monomer is preferably ≥ 99%. There is no strict requirement on the source, regular purchase is sufficient.

In the present invention, the dosage form of the drug is preferably a medically acceptable dosage form; the medically acceptable dosage form preferably includes any one of powder, injection, capsule, tablet and oral solution.

In the present invention, the inducement of gastric ulcer preferably includes alcohol and/or acid; the alcohol is preferably anhydrous ethanol; and the acid is preferably acetic acid.

Although the ergosterol of the present invention has a similar skeleton to corticosteroids, the effects of ergosterol and corticosteroids are different: corticosteroids increase gastric acid secretion, inhibit gastric mucus secretion, and promote the occurrence of gastric ulcers; ergosterol can promote the secretion of gastric mucus, resist gastric ulcers caused by anhydrous ethanol, and promote the recovery of experimental gastric ulcers caused by acetic acid. Ergosterol can effectively prevent and treat gastric ulcers. The results of the embodiment show that according to the dosage of 10-40 mg ergosterol/kg mouse and 50-500 mg ergosterol/d/person, gastric ulcers can be effectively prevented and treated.

The present invention also provides a drug for preventing and treating gastric ulcers, wherein the active ingredient of the drug includes ergosterol. In addition to being used directly alone, the ergosterol of the present invention also includes a drug with the function of preventing and treating gastric ulcers, which is composed of ergosterol as a raw material and an auxiliary material or auxiliary component allowed in the drug. The present invention has no strict requirements on the specific type of the auxiliary material, and it can be selected according to the conventional dosage form of the drug.

In order to further illustrate the present invention, the application of ergosterol provided by the present invention in the preparation of a drug for preventing and treating gastric ulcer is described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

Example 1

1. Main experimental materials

Ergosterol (purchased from McLean), lansoprazole (purchased from Jiangsu Renhe Pharmaceutical Co., Ltd.), 48 SD healthy rats, 200-220 g (purchased from Zhuhai Baishitong Biotechnology Co., Ltd.); 36 C57BL/6 healthy mice, 18-22 g (purchased from Guangzhou Ruige Biotechnology Co., Ltd.).

2. Animal Experiments

1. Prevention Experiment

(1) The healthy rats in step 1 were randomly divided into a control group, a model group, a lansoprazole group, a low-dose ergosterol group, a medium-dose ergosterol group, and a high-dose ergosterol group, with 8 healthy rats in each group. The rats were given medication according to the method in Table 1. During the medication period, the diet, drinking water and other conditions of the rats in each group were the same.

Table 1 Administration

(2) The drugs were continuously administered for 7 days according to Table 1. One hour after the last administration, the rats in the model group, lansoprazole group, low-dose ergosterol group, medium-dose ergosterol group, and high-dose ergosterol group were gavaged with 1 mL of anhydrous ethanol; the rats in the control group were gavaged with 1 mL of purified water. One hour after gavage, the rats were killed by overdose anesthesia, and the gastric tissues of the rats were obtained and dissected to observe the ulcers in the gastric tissues of the rats. The ulcer area was calculated by grading using ImageJ. The results are shown in Figure 1 and Table 2.

Table 2 Gastric ulcer area of rats in each test group (unit: mm ² , )

Note: “##” indicates P<0.01 compared with the control group; “**” indicates P<0.01 compared with the model group.

According to Table 2 and Figure 1, no gastric ulcer occurred in the rats in the control group, while large-area ulcers appeared in the stomach of the rats in the model group, indicating that the model was successfully constructed; lansoprazole can effectively alleviate gastric ulcers in rats induced by anhydrous ethanol; the gastric ulcer area of rats in the high-dose and medium-dose ergosterol groups was also significantly reduced, indicating that ergosterol has an effect on resisting acute gastric ulcers induced by anhydrous ethanol and can prevent ulcer disease.

(II) Treatment Experiments

(1) Model establishment

The healthy mice in step 1 were randomly divided into a control group and an experimental group, with 6 healthy mice in the control group and 30 healthy mice in the experimental group. The abdominal cavity of the control group was opened and randomly closed without treatment; the abdominal cavity of the experimental group was opened to expose the whole stomach, and acetic acid was dipped into a 2 mm diameter glass capillary, which was then poked into the stomach body for a total of 20 seconds. The abdominal cavity was closed after waiting for 1 minute. After the stomach was opened, a whitish ulcer surface could be seen, indicating that the gastric ulcer model was successfully constructed.

(2) On the second day after the model was established, the experimental mice were randomly divided into 5 groups, namely, model group, lansoprazole group, low-dose ergosterol group, medium-dose ergosterol group and high-dose ergosterol group, with 6 mice in each group. The drugs were administered according to the method in Table 3. During the drug administration period, the diet, drinking water and other conditions of each group of rats were the same.

Table 3 Administration

(3) After continuous administration for 3 days according to Table 3, the mice in each test group were killed, and the gastric tissues of the mice were obtained and dissected to observe the ulcers in the gastric tissues of the mice. The ulcer area was calculated by grading using ImageJ. The results are shown in Figure 2 and Table 4.

Table 4 Gastric ulcer area of mice in each test group (unit: mm ² , )

Note: “##” indicates P<0.01 compared with the control group; “**” indicates P<0.01 compared with the model group; and “*” indicates P<0.05 compared with the model group.

According to Figure 2 and Table 4, the mice in the control group had no gastric ulcers, while the model group had large-area gastric ulcers, indicating that the model was successfully constructed. Lansoprazole administration for 3 days did not significantly reduce the ulcer area, while high, medium and low doses of ergosterol could significantly repair gastric ulcers caused by acetic acid and effectively treat gastric ulcers.

Example 2

The Glide software was used to study the docking of the compound ergosterol with the RoRα protein, and the results were compared with the RoRα/Cholesterol cocrystal results. Molecular docking of ergosterol and RoRα protein was performed, and the results are shown in Figure 3. Among them, A in Figure 3 is the binding mode of RoRα with the substrate small molecule Cholesterol (cholesterol) (PDBID: 1N83), and B is the binding mode of RoRα with the substrate small molecule Ergosterol. The left picture in A and B is the overall view, and the right picture is a partial view. In the figure, the orange stick is a small molecule, the blue cartoon is a protein, the green dotted line represents the water bridge effect, and the gray dotted line represents the hydrophobic effect.

As can be seen from Figure 3, ergosterol and cholesterol bind to the same site on the RoRα protein and present very similar binding postures. From Figure B, we can observe that the small molecule Ergosterol forms a water bridge with R367 and Q289 on the RoRα protein. In addition, it also forms a hydrophobic interaction with amino acids such as V346, V379, I400, and I327 on the protein. Compared with the reference molecule Cholesterol, we found that Ergosterol forms a water bridge with R367 and R370 on the RoRα protein, and the hydrophobic interaction is similar to the RoRα/Ergosterol complex.

Example 3

Stability analysis

The root mean square deviation of molecular dynamics simulation can reflect the movement process of the complex. The larger the RMSD and the more violent the fluctuation, the more violent the movement. Conversely, the movement is stable. The binding stability of RoRα protein and ergosterol was predicted using Amber18 software, and the results are shown in Figure 4.

As shown in Figure 4, the RMSD fluctuations of the complexes of Ergosterol and cholesterol bound to RoRα protein during the simulation process were all within 3 angstroms, indicating that the small molecule binds very closely to the protein and the small molecule binding stability is very good. In addition, the RoRα/Ergosterol complex is not much different from the RoRα/Cholesterol complex, suggesting that Ergosterol has the potential for biological activity on RoRα protein. Activating RoRα protein can To promote the aggregation of M2 macrophages and repair gastric ulcers. As can be seen from Figure 4, ergosterol has the potential to activate RoRα, so ergosterol can prevent and treat gastric ulcers.

Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

Application of ergosterol in the preparation of medicines for preventing and treating gastric ulcer.
The use according to claim 1, characterized in that ergosterol is the only active ingredient.
The use according to claim 1 is characterized in that the cause of gastric ulcer includes alcohol and/or acid.
The use according to claim 3, characterized in that the alcohol is anhydrous ethanol.
The use according to claim 3, characterized in that the acid is acetic acid.
The use according to claim 1 is characterized in that the target of the drug is Rorα protein.
The use according to claim 6 is characterized in that the prevention and treatment of gastric ulcers includes promoting gastric mucus secretion and/or activating gastric stem cells.
The use according to any one of claims 1 to 7 is characterized in that the medicine comprises any one of powder, injection, capsule, tablet, pill, granule and oral solution.
A medicine for preventing and treating gastric ulcer, characterized in that the medicine comprises ergosterol and medically acceptable excipients.
The drug according to claim 9, characterized in that ergosterol is the only active ingredient of the drug.
A method for preventing and treating gastric ulcer, characterized in that ergosterol is taken at a dosage of 50 to 500 mg ergosterol/d/person.