CN107648242B

CN107648242B - Pharmaceutical composition for relieving and treating inflammatory bowel disease

Info

Publication number: CN107648242B
Application number: CN201710996749.XA
Authority: CN
Inventors: 亓云鹏; 段更利; 王仁萍; 张天; 陆峰; 宋云龙
Original assignee: Fudan University; Second Military Medical University SMMU
Current assignee: Fudan University; Second Military Medical University SMMU
Priority date: 2017-10-19
Filing date: 2017-10-19
Publication date: 2020-01-21
Anticipated expiration: 2037-10-19
Also published as: CN107648242A

Abstract

The invention provides a pharmaceutical composition for relieving and treating inflammatory bowel diseases, which is characterized by comprising tripterine and indoleamine-2, 3-dioxygenase inhibitors. The pharmaceutical composition provided by the invention comprises tripterine and an IDO1 inhibitor, wherein the tripterine and the IDO1 inhibitor respectively have certain effects of relieving and treating inflammatory bowel diseases, and the combination of the tripterine and the IDO1 inhibitor can further improve the curative effect on the basis of separate application, so that the pharmaceutical composition is an effective pharmaceutical composition for relieving and treating inflammatory bowel diseases, and can be used for relieving and treating inflammatory bowel diseases including Crohn's disease and ulcerative colitis.

Description

Pharmaceutical composition for relieving and treating inflammatory bowel disease

Technical Field

The invention belongs to the field of medicines, and relates to a pharmaceutical composition for relieving and treating inflammatory bowel disease.

Background

Inflammatory Bowel Disease (IBD) is a chronic recurrent intestinal disease including Crohn's Disease (CD) and Ulcerative Colitis (UC), with the main symptoms of diarrhea, abdominal pain, hematochezia, etc. The pathogenesis of IBD is not well defined and is thought to be related to factors such as environmental, microbial infection, genetic and immune system disorders. Anti-inflammatory drugs, corticosteroids, immunosuppressants and immunomodulators are currently common therapeutic drugs (such as mesalamine), however, most of the drugs have serious toxic and side effects and are not easy to cure. Therefore, research on the basis and application of novel drugs for treating IBD is attracting attention.

Tripterine (Celastrol) is a triterpenoid active monomer with a quinone methyl structure extracted from Tripterygium wilfordii hook.f., and is considered to be one of the most important active monomers in Tripterygium wilfordii. As a natural proteasome inhibitor, tripterine can play a pharmacological role in various inflammatory and autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus and dermatitis.

Research shows that tripterine has good application prospect in treating IBD: administration of tripterine to mice can effectively alleviate IBD models induced by Dextran Sodium Sulfate (DSS) and Trinitrobenzene sulfonic acid (TNBS); the tripterine can obviously reduce the levels of inflammatory mediators TNF alpha, IL-1 beta, IL-6 and IL-8 in intestinal mucosa of a Crohn disease patient; quantitative PCR and confocal microscopy analysis show that tripterine can inhibit LPS-mediated nuclear release of NF-kB in human monocytic cell lines.

Indoleamine-2, 3-dioxygenase (IDO1) is the only rate-limiting enzyme for catalyzing the catabolism of tryptophan along the canine uric acid pathway, and the expression of the rate-limiting enzyme is increased in IBD patients, and the rate-limiting enzyme can be used as an important index for judging the inflammation degree of the IBD patients. In recent years, IDO1 has been found^-/-The degree of inflammation of the gene-deleted mice is lower than that of IDO1^+/+Mice, and colitis mice were administered IDO1 inhibitor 1The methyl-tryptophan can obviously relieve the colitis of mice, which indicates that the IDO1 plays an important role in inflammatory response and regulation of a transcription network in the development process of the colitis, and also indicates that the IDO1 can be used as a target point for treating the colitis.

Therefore, the tripterine and the IDO1 inhibitor have treatment effects on IBD, the tripterine and the IDO1 inhibitor have the effects of inhibiting the generation of inflammatory mediators and inhibiting tryptophan catabolism pathway rate-limiting enzymes. Because the two drugs act in different ways and principles and the action mechanisms of the two drugs are complex, it is not clear whether the two drugs can be used for treating IBD in combination. In addition, the effect of the drugs with similar effects is easily reduced by antagonism when the drugs are combined, and whether the effect of the two drugs is reduced when the drugs are combined is not clear at present.

Disclosure of Invention

In order to solve the problems, the inventor of the invention explores the treatment effect of the tripterine and the IDO1 inhibitor in a colitis model mouse, finds that the combination of the tripterine and the IDO1 inhibitor can relieve the inflammatory symptoms of IBD, and the combination effect is better than the single application of the tripterine or the IDO1 inhibitor. Thus, the combination of tripterine and IDO1 inhibitors can be used to alleviate and treat IBD, and the effect of the combination is better than that of the individual use.

Based on the above findings, in order to provide a pharmaceutical composition with better therapeutic effect on IBD, the present invention adopts the following technical scheme: the invention provides a pharmaceutical composition for relieving and treating inflammatory bowel diseases, which is characterized by comprising tripterine and indoleamine-2, 3-dioxygenase inhibitors.

The pharmaceutical composition provided by the invention can also have the following technical characteristics, wherein the indoleamine-2, 3-dioxygenase inhibitor is 1-methyl-tryptophan.

The pharmaceutical composition provided by the invention also has the following technical characteristics, wherein the mass ratio of the tripterine to the 1-methyl-tryptophan is 1: 50.

The invention also provides the use of a pharmaceutical composition as described in any one of the above in the alleviation and treatment of inflammatory bowel disease.

The application of the pharmaceutical composition provided by the invention in relieving and treating inflammatory bowel disease can also have the following technical characteristics, wherein the inflammatory bowel disease is ulcerative colitis or Crohn's disease.

Action and Effect of the invention

The pharmaceutical composition provided by the invention comprises tripterine and an IDO1 inhibitor, and the combination of the tripterine and the IDO1 inhibitor can further improve the curative effect on the basis of single application, so that the pharmaceutical composition is an effective pharmaceutical composition for relieving and treating inflammatory bowel diseases, and can be used for relieving and treating inflammatory bowel diseases including Crohn's disease and ulcerative colitis.

Drawings

FIG. 1 is a graph of the change in body weight of mice over the course of a 7 day experiment in accordance with an embodiment of the present invention;

FIG. 2 is a histogram of colon length of mice after the experiment of the example of the present invention is completed;

FIG. 3 is a graph of the scoring of diarrhea in mice according to an embodiment of the present invention;

FIG. 4 is a graph of H & E staining of mouse colon tissue according to an embodiment of the present invention.

Detailed Description

The following examples are given to illustrate specific embodiments of the present invention. In the following examples, the reagents used were obtained from general commercial sources unless otherwise specified, and the experimental conditions not specified were referred to the conventional experimental conditions or the conditions recommended by the supplier. In addition, the IDO inhibitor used in the examples below was 1-methyl-tryptophan.

< example >

In the embodiment, dextran sulfate (DSS) is adopted to induce mice to generate colitis symptoms, and then the mice with colitis are administered in groups, and the remission and treatment effect of each group of drugs is examined by taking the remission condition of the inflammation of the mice after administration as an index.

The experimental process is as follows: the method comprises the following steps of randomly dividing mice into 6 groups, wherein each group comprises 5 mice, and adopting different treatment modes:

(1) the first group was the normal group (Water group): during the experiment (7 days), free drinking water was given, and no drug was added to the diet;

(2) the second group was the model group (DSS group): mice were given 3% DSS water for free drinking for 7 days;

(3) the third group was a positive control group, i.e., mesalamine administration group (ME + DSS): mice were given 3% DSS in water for free drinking for 7 days, while mesalamine (i.e. ME, dose 100mg/Kg) was administered once daily for 7 consecutive days by gavage;

(4) the fourth group is tripterine administration group (DSS + Celastrol): mice were given 3% DSS aqueous solution for free drinking, and simultaneously given tripterine (1mg/Kg) by gavage once a day for 7 consecutive days;

(5) the fifth group was an IDO Inhibitor administration group (DSS + IDO Inhibitor): mice were given 3% DSS in water for free drinking, while gavage was given an IDO inhibitor (50mg/kg), once daily for 7 consecutive days;

(6) the sixth group is a tripterine + IDO Inhibitor administration group (DSS + IDO Inhibitor + Celastrol): mice were given 3% DSS aqueous solution for free drinking, and simultaneously given tripterine (1mg/Kg) + IDO inhibitor (50mg/Kg) intragastrically, once daily for 7 consecutive days.

In each of the above experimental groups, the body weight, diarrhea and bleeding status of the mice were recorded before daily administration, and a score (0-4 points) was used as a measure of the severity of diarrhea (including bleeding of diarrhea).

After the experiment on the 7 th day is finished, the operation is not performed for 12 hours, blood is collected from orbital veins of each group of mice, and serum samples are collected. The mice were then sacrificed and after sacrifice the colons were removed quickly and measured for length, and after washing with physiological saline, fixed in 10% neutral buffered formalin for hematoxylin and eosin (H & E) staining detection.

Fig. 1 is a graph of the change in body weight of a mouse during a 7-day experiment in an example of the present invention, fig. 2 is a histogram of colon length of the mouse after the experiment in the example of the present invention is completed, fig. 3 is a score of diarrhea of the mouse in the example of the present invention, and fig. 4 is a graph of H & E staining of colon tissue of the mouse in the example of the present invention.

The mouse body weight, the mouse colon length and the mouse diarrhea score in fig. 1, fig. 2 and fig. 3 are all the mean values of the corresponding results in each experimental group, and the staining chart in fig. 4 is the staining chart obtained by any one staining specimen in the corresponding experimental group.

In fig. 1, the abscissa represents time, and the ordinate represents the body weight of the mouse. Significance between experimental groups was expressed using the P-value test. Wherein P <0.01, P <0.05, represents DSS group vs normal group; + P <0.01, representing DSS + celltrol group vs DSS group; # P <0.05, # P <0.01, representing the DSS + IDO inhibitor + celltrol group vs DSS group.

In fig. 2, the abscissa represents each experimental group, and the ordinate represents the colon length. Significance between experimental groups was also expressed using the P-value test. Wherein P <0.01, represents DSS group vs normal group; # P <0.05, representing the DSS + IDO inhibitor + cellstrol group vs DSS group.

In fig. 3, significance between experimental groups is also represented using the P-value test. Wherein P <0.01, P <0.05, represents DSS group vs normal group; + P <0.01, representing DSS + celltrol group vs DSS group; # P <0.05, # P <0.01, representing the DSS + IDO inhibitor + celltrol group vs DSS group.

Fig. 4(a) to 4(F) in fig. 4 correspond to the first to sixth groups, respectively.

Further, rectal bleeding of the mice in each of the above experimental groups is shown in table 1 below. In the column of "number of bleeding/total amount" in table 1, the number before the oblique line represents the number of mice in which rectal bleeding occurred, and the number after the oblique line represents the total number of mice.

TABLE 1 rectal bleeding in mice

As can be seen from fig. 1 to fig. 4 and table 1, compared with the normal group, the mice in the DSS group showed significant symptoms of weight loss, diarrhea, rectal bleeding, bloody stool, colon shortening and damaged colon epithelial cells, indicating that DSS can effectively induce mice to develop colitis symptoms, and all the indicators thereof conform to the symptoms and development rules of colitis.

Compared with the DSS group, the mesalazine administration group can relieve the weight reduction of mice, reduce the degree of colon shortening, rectal bleeding and diarrhea, and the results show that the existing colitis treatment drug can indeed have the effects of relieving and treating the DSS-induced colitis.

Meanwhile, compared with the DSS group, the tripterine administration group and the IDO inhibitor administration group can also relieve the weight reduction of mice, and reduce the degree of colon shortening, rectal bleeding and diarrhea. The effect of relieving and treating by independently applying the IDO inhibitor is close to that of mesalazine, and the effect of relieving and treating by independently applying the tripterine is better than that of the mesalazine.

Compared with the experimental groups, the administration group of the IDO inhibitor and the tripterine has better effects of relieving the weight reduction of mice, and reducing colon shortening, rectal bleeding and diarrhea. That is, when the IDO inhibitor and the tripterine are used together, the effect is reduced without antagonism, but the effect is better than that when the IDO inhibitor and the tripterine are used separately, and the effect is better than that of the mesalazine in the existing common medicine.

Examples effects and effects

The pharmaceutical composition provided according to this embodiment comprises tripterine and an IDO1 inhibitor, which can be used together to further improve the therapeutic effect on the basis of their single use, and thus is an effective pharmaceutical composition for alleviating and treating inflammatory bowel diseases, including crohn's disease and ulcerative colitis.

In the embodiment, the group of the IDO inhibitor and the tripterine shows better relieving and treating effects than two single medicines or mesalazine, which shows that the two medicines are combined to generate a certain synergistic effect and have better curative effect than the single medicine.

Claims

1. A pharmaceutical composition for the relief and treatment of inflammatory bowel disease, comprising tripterine and 1-methyl-tryptophan.

2. The pharmaceutical composition of claim 1, wherein:

wherein the mass ratio of the tripterine to the 1-methyl-tryptophan is 1: 50.

3. Use of a pharmaceutical composition according to any one of claims 1-2 for the preparation of a pharmaceutical formulation for the alleviation and treatment of inflammatory bowel disease.

4. Use of the pharmaceutical composition of claim 3 for the preparation of a pharmaceutical formulation for the amelioration and treatment of inflammatory bowel disease, wherein the inflammatory bowel disease is ulcerative colitis or Crohn's disease.