CN108354920B

CN108354920B - Application of sulfoxide compound in treating inflammatory bowel disease

Info

Publication number: CN108354920B
Application number: CN201810024040.8A
Authority: CN
Inventors: 饶子和; 杨诚; 孙涛; 白翠改; 赵秀娟; 孙桐艳; 李慧影; 刘艳荣; 路俊; 李晓慧; 陈璐
Original assignee: TIANJIN INTERNATIONAL JOINT ACADEMY OF BIOMEDICINE
Current assignee: TIANJIN INTERNATIONAL JOINT ACADEMY OF BIOMEDICINE
Priority date: 2018-01-10
Filing date: 2018-01-10
Publication date: 2020-06-16
Anticipated expiration: 2038-01-10
Also published as: CN108354920A

Abstract

The invention provides an application of sulfoxide compound in treating inflammatory bowel disease, the sulfoxide compound has a structure shown in the following general formula (I),

the sulfoxide compound has therapeutic effect on mice in acute stage of ulcerative colitis, chronic stage of ulcerative colitis and Crohn's disease, and can be used in medicine for treating or preventing inflammatory bowel disease.

Description

Application of sulfoxide compound in treating inflammatory bowel disease

Technical Field

The invention relates to the technical field of pharmacy, in particular to application of a sulfoxide compound in treating inflammatory bowel diseases.

Background

Inflammatory Bowel Disease (IBD) is a chronic nonspecific intestinal inflammatory disease, which has repeated courses of disease, and is clinically manifested by symptoms such as abdominal pain, diarrhea, mucopurulent bloody stool, tenesmus and the like, and has the pathological characteristics of chronic inflammation of the colonic mucosa, ulcer of the intestinal mucosa or formation of transmural granuloma. Epidemiological studies have found that IBD is more prevalent in north america and northern europe, but in recent years the prevalence has escalated in the asia-pacific region, with china becoming one of the countries with higher asia prevalence. IBD is better in young and strong years, has an average age of 39 years of onset, is more frequent in men, has a proportion of as high as 59 percent, and is inflammatory bowel disease or a mainstream digestive tract disease of Asian people in the next 10 years. Ulcerative Colitis (UC) and Crohn's Disease (CD) are the two major types of IBD, with CD occurring throughout the digestive tract, usually in the terminal ileum and right-half colon, with lesions affecting the entire layer of the intestinal wall, presenting as a segmental, thrush-like ulcer with necrotic granuloma. UC is frequently found in rectum and sigmoid colon, and the pathological changes involve mucous layer and submucosa, and show continuous and diffuse ulcer with congestion and edema and high recurrence. In addition, patients with UC have a significantly increased incidence of colorectal cancer (CRC), which is approximately 20 times higher than the general population. Over 20% of patients with UC develop colitis-associated colon cancer (CAC) within 30 years of diagnosis, with mortality rates of more than 50%.

IBD, a chronic intractable disease that cannot be cured, has an undefined pathogenesis and is generally considered to be caused by the interaction of various factors such as environment, heredity, intestinal flora and immune response. At present, IBD cannot be completely cured, no drug with strong specificity and small toxic and side effects exists, and the treatment of IBD is mainly based on inflammation control, immune function regulation, attack control and remission maintenance. Traditional therapeutic drugs mainly include salicylic acid preparations, glucocorticoids and immunosuppressive agents. But has large side effect and high incidence rate, and can not be used for a long time. In biological preparations, TNF-t and monoclonal antibodies are novel drugs applied to IBD treatment in recent years, and show good effects in controlling disease symptoms and promoting mucosal healing, and currently, Infliximab (IFX) and adalimumab (adalimumab) are mainly applied, but only anti-TNF-t and a monoclonal antibodies are suitable for maintenance treatment.

Therefore, the method has important significance and social demand on developing a novel medicament with definite treatment effect and low toxic and side effect aiming at inflammatory bowel diseases, and has wide market prospect.

Disclosure of Invention

In view of the above problems, the present invention provides a sulfoxide compound for use in the treatment of inflammatory bowel disease, wherein the sulfoxide compound has the following general formula (I):

in the above applications, the inflammatory bowel disease includes ulcerative colitis and crohn's disease.

In the above use, the ulcerative colitis includes an acute phase of ulcerative colitis and a chronic phase of ulcerative colitis.

In the above application, the dosage of the sulfoxide compound is 5mg/Kg to 20 mg/Kg.

The present invention also provides a medicament for treating or preventing inflammatory bowel disease, comprising: sulfoxide compounds and pharmaceutically acceptable carriers.

In the above drugs, the pharmaceutically acceptable carrier includes one or more of diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants and synergists.

In the above drugs, the formulation of the drug includes injection, tablet, pill, capsule, suspension or emulsion.

According to the invention, the sulfoxide compound has a treatment effect on mice in acute stage of ulcerative colitis, mice in chronic stage of ulcerative colitis and mice with Crohn's disease, and the effect of the sulfoxide compound is superior to that of positive drug sulfasalazine (SASP), so that the sulfoxide compound can be applied to medicines for treating or preventing inflammatory bowel diseases.

Drawings

FIG. 1 is a graph of the body weight of mice in the acute phase of ulcerative colitis;

FIG. 2 is a graph of disease activity index in mice in the acute phase of ulcerative colitis;

FIG. 3 is a bar graph of colorectal length of mice in the acute phase of ulcerative colitis;

FIG. 4A is a bar graph of colorectal gross morphology score for mice in the acute phase of ulcerative colitis;

FIG. 5A is a bar graph of colorectal tissue damage score for mice in the acute phase of ulcerative colitis and FIG. 5B is a bar graph of colorectal tissue damage score;

FIG. 6 is a bar graph of colorectal length of mice in the chronic phase of ulcerative colitis;

FIG. 7A is a colorectal gross morphology of mice in chronic phase of ulcerative colitis, and FIG. 7B is a bar graph of colorectal gross morphology score;

FIG. 8A is a bar graph of colorectal tissue damage score for mice in chronic stage of ulcerative colitis and FIG. 8B is a bar graph of colorectal tissue damage score;

FIG. 9 is a bar graph of colorectal length in Crohn's disease mice;

FIG. 10 is a bar graph of colorectal gross morphology score for Crohn's disease mice;

figure 11 is a bar graph of colorectal tissue damage scores for crohn's disease mice.

Detailed Description

In order to better illustrate the present invention, specific embodiments thereof will be described in detail below.

The invention provides an application of a sulfoxide compound in treating inflammatory bowel diseases, wherein the sulfoxide compound has a structure shown in a general formula (I):

the following examples evaluate the efficacy of JY4 in three inflammatory bowel disease models, an acute phase model of ulcerative colitis, a chronic phase model of ulcerative colitis and a Crohn's disease model, respectively, but do not limit the invention in any way.

Study on therapeutic effect of JY4 on mice in acute stage of ulcerative colitis

1.1 Experimental methods

The selected animal and food system in the experiment is BALB/c mouse (white-variant laboratory mouse), female, 18-22 g, SPF (specific pathogen free) grade, and 48 tested medicines: compound JY 4. Positive control drug: sulfasalazine (SASP). Molding inducer: dextran Sodium Sulfate (DSS).

1.1.1 model building

DSS was used to induce the acute model of ulcerative colitis. During the adaptation period, the mice were given free food and water, and the experiment was started by changing the drinking water to 3-5% by mass DSS solution (excluding normal control group) and were given free water. The daily water intake of each mouse is calculated according to 6mL, sufficient DSS solution is supplemented to the daily water intake the next day, after the DSS solution is administered for 3-5 days, the phenomenon of soft stool, diarrhea or bloody stool of the mouse is found, the model is successfully established, and the DSS solution is continuously drunk until the experiment is finished.

1.1.2 grouping and administration

Namely, the mice were grouped into groups of 8 mice by weight, and each group contained a Normal control group (Normal), a Model control group (Model), a positive control group (SASP-200mg/kg), a drug administration group JY4(5mg/kg), a drug administration group JY4(10mg/kg), and a drug administration group JY4(20 mg/kg). The medicine is administrated by a gastric lavage way, about 0.1 mL. The dosage is respectively as follows: the normal control group and the model control group mice are given 0.1mL of medical saline every day, the positive control group mice are given 0.1mL of sulfasalazine-containing saline (200mg/kg) every day, and the administration group mice are respectively given 0.1mL of JY 4-containing saline (5mg/kg, 10mg/kg and 20mg/kg) every day. The general living state, fecal characteristics, and diarrhea and bloody stool of the mice were observed daily and the body weights were weighed. All mice were euthanized 7 days after dosing, the abdominal cavity of the mice was exposed, the colorectal was dissected and the length was measured; the colon and rectum were cut longitudinally along the mesentery, and the feces were washed with physiological saline and fixed with 10% neutral formalin solution. And (3) observing the general damage by using a stereoscopic microscope, grading, taking the severe ulcer to perform pathological examination, sequentially performing dehydration, paraffin embedding, slicing and HE staining, and observing the pathological condition of the tissue under the microscope and grading.

1.2. Results of the experiment

(1) Influence of JY4 on body weight of mice in acute stage of ulcerative colitis

As can be seen in FIG. 1, the weight of each dosage group of JY4 and the weight of the positive control group (SASP-200mg/kg) are larger than that of the model control group; compared with the positive control group, the weight of each dose group of JY4 is larger than that of the positive control group, and the difference has statistical significance (P < 0.05). The result shows that JY4 has obvious attenuation effect on the weight loss of the ulcerative colitis in the acute stage, and the effect is superior to that of a positive medicine SASP.

(2) Influence of JY4 on disease activity index of mice in acute stage of ulcerative colitis

The normal control group mice have active behaviors, bright hair color and smooth fur; the mice in the model control group are cachectic and do not move slowly, and the coat is rough and lusterless, while the mice in the positive control group and the JY4 low, middle and high groups generally have good activity, smooth coat, bright hair color and improved appetite. The mouse disease activity index was scored for weight loss and fecal status as shown in figure 2.

As can be seen from FIG. 2, compared with the normal control group, the disease activity index score of the mice in the model control group is obviously increased, and the differences have statistical significance (P < 0.05); compared with a model control group, the disease activity index scores of each dosage group of JY4 and the positive control group are obviously reduced, and the differences have statistical significance (P < 0.05); compared with a positive control group (SASP-200mg/kg), the disease activity index scores of the JY4 high-dose group are lower than those of the positive control group, the disease activity index scores of the JY4 middle-and-low-dose groups are equivalent to those of the positive control group, and the differences are statistically significant (P < 0.05). The result shows that JY4 can obviously reduce the disease activity index score of the ulcerative colitis mouse in the acute stage, and the effect is superior to that of a positive medicine SASP.

(3) Influence of JY4 on colorectal length of mice in acute stage of ulcerative colitis

As can be seen from FIG. 3, the colorectal cancer of the model control group was significantly shortened as compared with the normal control group; compared with a model control group, the colorectal lengths of the JY4 low, medium and high dose groups are obviously longer than those of the model control group, and in addition, the colorectal lengths of the mouse colorectal of the positive control group (SASP-200mg/kg) are also obviously longer than those of the model control group, but are obviously shorter than those of the JY4 medium and high dose groups; comparing the colorectal length between the JY4 doses, the colorectal length was significantly longer in the middle and high dose groups than in the low dose group (P < 0.05). Results show that JY4 can obviously restore the colorectal length of mice in the acute stage of ulcerative colitis, and the effect of medium and high dose groups is obviously superior to that of a positive medicine SASP.

(4) Influence of JY4 on colorectal gross morphology of mice in acute stage of ulcerative colitis

Gross morphology observation of mouse colon mucosa as shown in fig. 4A, the mucosa of normal control mice was smooth and regular. In the acute stage model of ulcerative colitis, the plica of the mucous membrane of a mouse in a model control group disappears, the mucous membrane is subjected to diffuse congestion, edema and vascular texture disorder, a large amount of mucus or purulent blood secretion exists in the intestinal cavity, erosion and ulcer can be formed, the ulcer surface is large, the boundary is clear, the inflammatory reaction is obvious, and the intestinal wall of some mice is thickened and adhered to the surrounding tissues; the colorectal gross lesion of mice in each dose group of a positive control group (SASP-200mg/kg) and JY4 is obviously lighter than that of a model group, and particularly the recovery effect of the middle and high dose groups of JY4 is more obvious.

The gross morphology scoring results are shown in fig. 4B, with significantly higher model control group scoring values; the positive control group and JY4 have low, medium and high dose groups with scores obviously lower than those of the model control group; the lesion scores of the medium and high dose groups are obviously lower than those of the positive control group; when JY4 is compared among the dose groups, the scores of the middle and high dose groups are obviously lower than those of the low dose group, and the differences have statistical significance (P < 0.05). Results show that JY4 can obviously recover mucosal ulcer condition in the acute stage of ulcerative colitis, and the effect of the medium and high dose groups is obviously superior to that of the positive drug SASP.

(5) Influence of JY4 on mouse colorectal tissue injury in acute stage of ulcerative colitis

The observation of mouse colon mucosa tissue is shown in fig. 5A, the normal control group mouse colon tissue structure is regular, the mucosa layer, submucosa, muscularis layer and serosa layer are all complete, the colon structure is clear, the glands are arranged regularly, and less inflammatory cell infiltration can be seen. The damage of the colon mucous membrane of the mouse in the model control group is serious, the mucous membrane epithelial cells are fallen off, the arrangement of gland bodies in the mucous membrane is disordered, deformed or even lost, the crypt abscess is formed, congestion and edema are accompanied, and a large amount of neutrophil granulocytes and lymphocyte infiltration can be seen. Positive control group (SASP-200mg/kg) and JY4 low dose group have mucosal epithelium exfoliation, mild hyperemia and edema and weakened inflammatory cell infiltration; JY4 middle and high dose group mouse lesion is obviously reduced, ulcer is healed, mucous membrane is complete, and inflammatory cell infiltration is obviously reduced compared with a model group.

The tissue damage score results are shown in fig. 5B, where the colorectal tissue damage score values of the model control group were significantly higher than that of the normal control group; compared with a model control group, the tissue damage scores of the JY4 low, medium and high doses and the positive control group are all obviously reduced; compared with a positive control group, the scores of the JY4 medium and high dose groups are obviously reduced; when the JY4 is compared among the dose groups, the scores of the middle and high dose groups are obviously lower than those of the low dose group, and the difference has statistical significance (P < 0.05). Results show that JY4 can obviously relieve colorectal tissue injury of mice in the acute stage of ulcerative colitis, reduce mucosal exfoliation necrosis and inflammatory cell infiltration degree, and the effect of medium and high dose groups is obviously superior to that of a positive drug SASP.

Study on therapeutic effect of (II) JY4 on mice in chronic stage of ulcerative colitis

The test selects 48 test drugs with the animal strains of BALB/c mice, female, 18-22 g and SPF grade: compound JY 4. Positive control drug: sulfasalazine (SASP). Molding inducer: dextran Sodium Sulfate (DSS).

2.1. Experimental methods

2.1.1 model building

A chronic model of DSS induced ulcerative colitis was used. During the acclimation period, the mice were given free food and water, and the experiment was started by changing the drinking water to 3% by mass DSS solution (excluding the normal control group) and were given free water. The daily water intake of each mouse is calculated according to 6mL, sufficient DSS solution is supplemented to the daily water intake the next day, drinking water is used after the DSS solution is given for 7 days in total, and after the DSS solution is freely drunk for 7 days, the drinking water is recycled for a period according to the method, and the modeling is successful. The above cycle was followed until the end of the experiment.

2.1.2 grouping and administration

Consistent with 1.1.2 method; the administration time was 14 days.

2.2. Results of the experiment

(1) Influence of JY4 on colorectal length of mice in chronic stage of ulcerative colitis

JY4 has an effect on the colorectal length of mice in the chronic stage of ulcerative colitis as shown in FIG. 6, and as can be seen from FIG. 6, compared with a normal control group, the colorectal length of a model control group is obviously shortened; compared with a model control group, the colorectal lengths of the JY4 low, medium and high dose groups are obviously longer than those of the model control group, and in addition, the colorectal lengths of the mouse colorectal of the positive control group (SASP-200mg/kg) are also obviously longer than those of the model control group, but are obviously shorter than those of the JY4 medium and high dose groups; comparing the colorectal length between JY4 doses, the colorectal length of the middle and high dose groups is obviously longer than that of the low dose group, and the difference has statistical significance (P < 0.05). Results show that JY4 can obviously restore the colorectal length of mice in the chronic stage of ulcerative colitis, and the effect of medium and high dose groups is obviously superior to that of a positive medicine SASP.

(2) Influence of JY4 on colorectal gross morphology of mice in chronic stage of ulcerative colitis

Gross morphology observation of mouse colon mucosa as shown in FIG. 7A, the mucosa of normal control mice was smooth and regular. In the acute-stage model of ulcerative colitis, the plica of the mucous membrane of a mouse in a model control group disappears, the mucous membrane is subjected to diffuse congestion and edema, the vascular texture is disordered, a large amount of mucus or purulent blood secretion exists in the intestinal cavity, erosion and ulcer can be formed, the ulcer surface is large, the boundary is clear, and the inflammatory reaction is obvious; the colorectal gross lesions of mice in positive control groups and JY4 dose groups are obviously lighter than those in model groups, and particularly the recovery effect of JY4 middle and high dose groups is more obvious.

The gross morphology scoring results are shown in fig. 7B, with significantly higher model control group scoring values; the positive control group and JY4 have low, medium and high dose groups with scores obviously lower than those of the model control group; the lesion scores of the medium and high dose groups are obviously lower than those of the positive control group, the scores of the medium and high dose groups are obviously lower than those of the low dose group by comparison among the JY4 dose groups, and the differences have statistical significance (P < 0.05). Results show that JY4 can obviously recover mucosal ulcer condition in chronic stage of ulcerative colitis, and the effect of medium and high dose groups is obviously superior to that of positive drug SASP.

(3) Influence of JY4 on mouse colorectal tissue injury in chronic stage of ulcerative colitis

The observation of mouse colon mucosa tissue is shown in fig. 8A, the normal control group mouse colon tissue structure is regular, the mucosa layer, submucosa, muscularis layer and serosa layer are all complete, the colon structure is clear, the glands are arranged regularly, and less inflammatory cell infiltration can be seen. The damage of the colon mucous membrane of the mouse in the model control group is serious, the mucous membrane epithelial cells are fallen off, the arrangement of gland bodies in the mucous membrane is disordered, deformed or even lost, the crypt abscess is formed, congestion and edema are accompanied, and a large amount of neutrophil granulocytes and lymphocyte infiltration can be seen. The positive control group and the JY4 low-dose group have mucosal epithelium shedding, mild hyperemia and edema and weakened inflammatory cell infiltration; JY4 middle and high dose group mouse lesion is obviously reduced, ulcer is healed, mucous membrane is complete, and inflammatory cell infiltration is obviously reduced compared with a model group.

The tissue damage score results are shown in fig. 8B, where the colorectal tissue damage score values of the model control group were significantly higher than that of the normal control group; compared with a model control group, the tissue damage scores of the JY4 low, medium and high doses and the positive control group are all obviously reduced; compared with a positive control group, the scores of the JY4 medium and high dose groups are obviously reduced; when the JY4 is compared among the dose groups, the scores of the middle and high dose groups are obviously lower than those of the low dose group, and the difference has statistical significance (P < 0.05). Results show that JY4 can obviously relieve colorectal tissue injury of mice in the chronic stage of ulcerative colitis, reduce mucosal exfoliation necrosis and inflammatory cell infiltration degree, and the effect of medium and high dose groups is obviously superior to that of a positive drug SASP.

Study on therapeutic effect of (tri) JY4 on Crohn disease mice

The test selects 42 animal strains of BALB/c mice, female, 18-22 g, SPF grade, and the total number of test drugs is as follows: compound JY 4. Positive control drug: sulfasalazine (SASP). Molding inducer: trinitrobenzenesulfonic acid (TNBS).

3.1 Experimental methods

3.1.1 model building

TNBS was used to induce a crohn's disease model. The mice were given free food and water, and the experiment was started by first intraperitoneally injecting a suitable amount of 10% (mass fraction) chloral hydrate lightly anesthetized mice, lubricated with a catheter of about 2mm in diameter, and inserted into the colon about 4cm from the anus. Normal control group was injected with 0.1mL of physiological saline per mouse, solvent control group was injected with 0.1mL of 50% ethanol solution per mouse, and the remaining mice were injected with an equal volume of 50% ethanol solution of 24mg/mL TNBS (120 mg/kg). Inverting tail end of herba Swertiae Mileensis for 5min, and placing into cage for continuous feeding after the medicine is completely absorbed. After 24h of clysis, the mice have the phenomena of soft stool, diarrhea or bloody stool, and the modeling is successful.

3.1.2 grouping and administration

Consistent with 1.1.2 method; the administration time was 7 days.

3.2 analysis of the results

(1) Effect of JY4 on colorectal Length of Crohn's disease mice

Colorectal lesions were evaluated by measuring the colorectal length of mice, as shown in figure 9:

compared with a normal control group, the colorectal cancer of the model control group is obviously shortened; compared with a model control group, the colorectal lengths of the JY4 low, medium and high dose groups are obviously longer than those of the model control group, and in addition, the colorectal lengths of the mouse colorectal of the positive control group (SASP-200mg/kg) are also obviously longer than those of the model control group, but are obviously shorter than those of the JY4 medium and high dose groups; comparing the colorectal length between JY4 doses, the colorectal length of the middle and high dose groups is obviously longer than that of the low dose group, and the difference has statistical significance (P < 0.05). The result shows that JY4 can obviously restore the colorectal length of a Crohn disease mouse, and the effect of the medium and high dose groups is obviously superior to that of a positive drug SASP.

(2) Influence of JY4 on colorectal gross morphology of Crohn's disease mice

Observing the gross morphology of the colorectal mucosa of the mouse, and carrying out gross morphology scoring, wherein the scoring result is shown in figure 10: compared with a normal control group and an ethanol control group, the score value of the model control group is obviously higher; the positive control group and JY4 have low, medium and high dose groups with scores obviously lower than those of the model control group; the lesion scores of the low, medium and high dose groups are obviously lower than those of a positive control group (SASP-200mg/kg), and compared among the JY4 dose groups, the low, medium and high dose groups have dose dependence, and the difference has statistical significance (P < 0.05). The result shows that JY4 can obviously recover the mucosal ulcer condition of a Crohn disease mouse, and the effect is obviously superior to that of a positive medicine SASP.

(3) Effect of JY4 on Crohn's disease mouse colorectal tissue injury

Mouse colonic mucosal tissue was observed and scored for tissue damage, with the scoring results shown in fig. 11:

the result shows that the colorectal tissue damage score of the model control group is obviously higher than that of the normal control group; compared with a model control group, the tissue damage scores of the JY4 low, medium and high doses and the positive control group are all obviously reduced; compared with a positive control group, the scores of the JY4 medium and high dose groups are obviously reduced; when the JY4 is compared among the dose groups, the scores of the middle and high dose groups are obviously lower than those of the low dose group, and the difference has statistical significance (P < 0.05). Results show that JY4 can obviously reduce colorectal tissue injury of mice with Crohn's disease, reduce mucosal exfoliation necrosis and inflammatory cell infiltration degree, and the effect of medium and high dose groups is obviously superior to that of a positive drug SASP.

In conclusion, JY4 (sulfoxide compound) has treatment effect on mice in acute stage of ulcerative colitis, chronic stage of ulcerative colitis and Crohn's disease, and the effect of the medium and high dose groups is obviously better than that of the positive drug SASP.

Furthermore, JY4 and a pharmaceutically acceptable carrier can be prepared into a medicament for treating or preventing inflammatory bowel diseases, wherein the acceptable carrier comprises one or more of diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants and synergists; and can be made into injection, tablet, pill, capsule, suspension or emulsion.

The present invention is not limited to the above embodiments, and any modifications, equivalent substitutions, improvements, etc. within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. Use of a sulfoxide compound in the manufacture of a medicament for the treatment of inflammatory bowel disease, wherein said sulfoxide compound has the following general formula (I):

2. the use according to claim 1, wherein the inflammatory bowel disease is ulcerative colitis and Crohn's disease.

3. The use according to claim 2, wherein the ulcerative colitis is ulcerative colitis in its acute phase and in its chronic phase.

4. The use according to claim 1, wherein the dose of the sulphoxide compound is from 5mg/Kg to 20 mg/Kg.