CN111991387B - Application of dehydrodiisobutyrol in preparing medicine for preventing and treating ulcerative colitis - Google Patents

Abstract

The invention relates to the field of medicine and pharmacology, in particular to a new medical application of dehydrodiisobutyronitrile. The dehydrodiisobutyronitrile can be used for preparing medicines for preventing and treating ulcerative colitis.

Description

Application of dehydrodiisobutyrol in preparing medicine for preventing and treating ulcerative colitis

Technical Field

The invention relates to the field of medicines, in particular to a new application of dehydrodiisobutyro-eugenol.

Background

Ulcerative Colitis (UC) is an inflammatory disease of the intestinal tract with an unknown etiology and a high recurrence rate, and constitutes Inflammatory Bowel Disease (IBD) together with Crohn's Disease (CD). UC can occur in all age groups and is difficult to cure, and the incidence rate tends to increase remarkably in China, so that UC is one of modern difficult diseases. Chronic nonspecific inflammation mainly affecting colorectal mucosa due to pathological changes is mainly abdominal pain, diarrhea, mucous bloody stool and tenesmus, is delayed in course of disease, is easy to recur, and has the possibility of canceration. At present, the treatment of UC is mainly based on western medicines, including steroids and non-steroids, such as glucocorticoids, aminosalicylates, antibiotics, immunosuppressants and the like. However, the long-term use of these drugs has large side effects and the drugs are likely to rebound after the drug is stopped, for example, clinical SASP can cause hepatotoxicity. Because Chinese medicine resources are rich, the new application of screening active new compounds or natural compounds with known structures from natural plants becomes one of the main approaches for developing new medicines. Therefore, it is an important subject in the art to find natural drugs for treating ulcerative colitis from natural plants.

Dehydrodiisobutyronitrile is an active ingredient mainly separated from nutmeg, and has antiinflammatory, antibacterial, antioxidant and antitumor activities. However, no report that dehydrodiisobutyronitrile can be used as the only active ingredient for preparing the medicament for preventing or treating ulcerative colitis is found at present.

Disclosure of Invention

The invention aims to provide a new medical application of dehydrodiisobutyronitrile.

Specifically, the invention provides an application of dehydrodiisobutyronitrile in preparing a medicament for preventing and treating ulcerative colitis.

In a preferable embodiment, the dehydrodiisobutyl phenol is used as the only active ingredient for preparing the medicine for preventing and treating ulcerative colitis.

The details of various aspects of the invention are set forth in subsequent sections. The features, objects, and advantages of the invention will be apparent from the description and from the claims.

Drawings

FIG. 1 shows the effect of dehydrodiisobutyronitrile on the body weight of mice with ulcerative colitis;

FIG. 2 shows the effect of dehydrodiisobutyronitrile on the incidence of bloody stools in mice with ulcerative colitis;

FIG. 3 shows the effect of dehydrodiisobutyronitrile on colon length in ulcerative colitis mice, FIG. 3A shows the overall appearance of the colon in mice, FIG. 3B shows the colon length in mice;

FIG. 4 shows the effect of dehydrodiisobutyronitol on colon histopathology in mice with ulcerative colitis, FIG. 4A staining of HE in the colon of mice, and FIG. 4B histopathology scoring;

(in the above-mentioned figure: a represents a normal control group, b represents a model control group, c represents a SASP group, and d represents a dehydrodiisobutyronitrile group)

Detailed Description

The invention arose in part from the unexpected discovery that: the dehydrodiisobutyronitrile can obviously improve the symptoms of weight loss, bloody stool, colon shortening, histopathological injury and the like of experimental ulcerative colitis mice. Therefore, the dehydrodiisobutyrol can be used for preparing the medicine for preventing and treating enteritis, particularly ulcerative colitis.

Furthermore, the invention provides the application of the dehydrodiisobutyronitrile in preparing the medicine for preventing and treating the ulcerative colitis.

The molecular formula of the dehydrodiisobutyl phenol is as follows: c₂₀H₂₂O₄The molecular weight is: 326.4, having the formula:

the dehydrodiisobutyronitrile of the present invention can be commercially obtained from Vickers Biotech Co., Ltd, Chengdu Manster Biotech Co., Ltd, Shanghai pure superior Biotech Co., Ltd, etc., of Sichuan province. The purity of the product meets the pharmaceutical standard. The purity of the dehydrodiisobutyl phenol of the invention is more than or equal to 97 percent.

The dehydrodiisobutyrol of the present invention may be used alone or in the form of a pharmaceutical composition. The pharmaceutical composition comprises the dehydrodiisobutyronitrile of the invention as an active ingredient and a pharmaceutically acceptable carrier. Preferably, the pharmaceutical composition of the present invention contains 0.1 to 99.9% by weight of the dehydrodiisobutyl eugenol of the present invention as an active ingredient. The pharmaceutical carrier does not destroy the pharmaceutical activity of the dehydrodiisobutyl phenol, and the effective dosage of the dehydrodiisobutyl phenol is nontoxic to human bodies.

Such pharmaceutically acceptable carriers include, but are not limited to: lecithin, aluminum stearate, alumina, ion exchange materials, self-emulsifying drug delivery systems, tweens or other surfactants, serum proteins, buffer substances such as phosphates, glycine, sorbic acid, water, salts, electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, magnesium silicate, mixtures of saturated fatty acid partial glycerides, and the like.

Other conventional pharmaceutical adjuvants such as binder (e.g. microcrystalline cellulose), filler (e.g. starch, glucose, anhydrous lactose and lactose beads), disintegrant (e.g. crosslinked PVP, croscarmellose sodium, low-substituted hydroxypropylcellulose), lubricant (e.g. magnesium stearate), and absorption enhancer, adsorption carrier, flavoring agent, sweetening agent, excipient, diluent, wetting agent, etc.

The dehydrodiisobutyrol and pharmaceutical compositions thereof of the present invention may be prepared according to conventional methods in the art and may be administered by enteral, parenteral, or topical routes. The oral preparation comprises capsule, tablet, oral liquid, granule, pill, powder, pellet, and unguent; parenteral preparations include injections and the like; formulations for topical administration include creams, patches, ointments, sprays, and the like. Oral formulations are preferred.

The administration route of the dehydrodiisobutyrol and the pharmaceutical composition thereof can be oral, sublingual, transdermal, intramuscular or subcutaneous, skin mucosa, vein, urethra, vagina and the like.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Experimental procedures without specific conditions noted in the following examples, generally according to conventional conditions or according to conditions recommended by the manufacturers. All percentages, ratios, proportions, or parts are by weight unless otherwise specified.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.

The features mentioned above with reference to the invention, or the features mentioned with reference to the embodiments, can be combined arbitrarily. All the features disclosed in this specification may be combined in any combination and each feature disclosed in this specification may be replaced by alternative features serving the same, equivalent or similar purpose. Thus, unless expressly stated otherwise, the features disclosed are merely generic examples of equivalent or similar features.

Example 1:

1. experimental Material

1.1 medicinal materials

Dehydrodiisobutyronitrile (CAS:2680-81-1, MW 326.4, HPLC purity not less than 98%, Vickers Biotech, Inc., Sichuan); dextran sulfate sodium (DSS, CAS: 9011-18-1, MW 36-50kDa, manufactured by MP Biomedicals, USA); sulfasalazine (SASP, CAS: 599-79-1, molecular weight 398.39, HPLC purity 98% or more, manufactured by Sigma-Aldrich Co., USA).

1.2 Experimental animals

C57BL/6J female mice weighing 20 ± 2g (8 weeks), provided by the experimental animals center of the university of medicine, shanghai, animal certification No.: SYXK (Shanghai) 2018-0032. Placing in conventional breeding environment, and freely taking food and drinking water.

2. Experimental methods

2.1 establishment of ulcerative colitis model

Female mice (20 + -2 g) of C57BL/6J, which were of uniform body weight, were selected and given free access to food and water at the beginning of the study using the International general ulcerative colitis modeling method (Gastroenterology 2002,123: 256-70; PLoS One 2012,7: e36075), and water was changed to 4% (w/v) DSS for 7 days after the initial acclimation phase to induce ulcerative colitis.

2.2 methods of grouping and administering drugs

Normal control group: 10, and normal diet.

Model control group: 10 were given 4% DSS solution for 7 days of free drinking.

SASP group: 10 of these, 4% DSS solution was given with SASP (350mg/kg) administered by gavage for 7 consecutive days.

Dehydrodiisobutylaminol group: 10 patients were administered 4% DSS solution simultaneously with gavage of dehydrodiisobutyl eugenol (50mg/kg) for 7 consecutive days.

During the experiment, dosage formulations were prepared fresh daily, dissolved in 0.5% sodium carboxymethyl cellulose and used within 1 hour of preparation.

2.3 evaluation of enteritis

Diarrhea, bloody stool, histopathological analysis, etc. were performed according to the methods described in the earlier published papers (Am J Physiol gastroenterest Liver Physiol,2015,309(7): G517-G527). Body weight changes, diarrhea and bloody stool symptoms were recorded daily during the experiment. After the experiment was completed, the mice were sacrificed by anesthesia and cervical dislocation, the abdominal cavity was opened, the colon was removed, and the length of the colon from the cecum to the rectum was measured. Taking the tail colon as a tissue section, carrying out H & E staining, and carrying out pathological scoring on the H & E stained colon specimen: (1) exudation scoring criteria for inflammatory cells: minute quantity of inflammatory cells exist in the 0 minute-mucous membrane inherent layer, more inflammatory cells exist in the 1 minute-mucous membrane inherent layer or the inflammatory cells in the mucous membrane inherent layer are increased, 2 minute-inflammatory cells are diffused to the submucosa, and 3 minute-whole layer has inflammatory cell exudation; (2) tissue damage scoring criteria: 0 point-no mucosal damage, 1 point-discontinuous mucosal epithelial damage, 2 points-superficial mucosal erosion, and 3 points-extensive mucosal damage and deep extension to intestinal wall. The exudation score and tissue damage score of inflammatory cells were added and a histopathological score (1-6 points) was calculated.

2.4 statistical analysis

All experimental data were repeated 3 times, the results were expressed as mean ± standard deviation, and One-way analysis of variance (One-way ANOVA) and LSD test were performed on the experimental data using SPSS 16.0 statistical software, with P <0.05 being statistically significantly different.

3. Results

3.1 therapeutic Effect of dehydrodiisobutyroninol on ulcerative colitis

3.1.1 Effect on body weight in mice with ulcerative colitis

FIG. 1 shows the effect of dehydrodiisobutyronitrile on the body weight of mice with ulcerative colitis. The body weight of the normal control group mice changed steadily throughout the experiment, and the body weight (20.6 + -0.2 g) increased by 0.6% on day 7; the body weight of the model group mice continuously decreased, and the weight loss rate on day 7 (14.4. + -. 0.3g) was 28.1%; the weight loss rates of mice in the SASP group (16.1 +/-0.2 g) and the dehydrodiisobutyronitrile group (17.4 +/-0.2 g) were 20.1% and 16.4%, respectively, which were significantly different from those in the model group (P <0.01 and P < 0.001). The SASP and the dehydrodiisobutyronitrile can obviously improve the weight loss symptom of mice with the ulcerative colitis caused by the DSS; and in the case of oral administration, the effect of suppressing the weight loss by the dehydrodiisobutyro-eugenol (50mg/kg) is obviously better than that of a positive control drug SASP (350mg/kg) (P is less than 0.05).

3.1.2 Effect on the incidence of bloody stools in mice with ulcerative colitis

FIG. 2 shows the effect of dehydrodiisobutyronitrile on the incidence of bloody stools in mice with ulcerative colitis. Throughout the experiment, normal control mice developed no blood. Starting on day 4, 3 groups of mice except the normal control group began to develop bloody stool symptoms, with the model group of mice being the most severe. In the model group, the patients are subjected to DSS water for 5-7 days, diarrhea and bloody stool are gradually serious, and the model group has obvious difference (P is less than 0.001) compared with a normal control group; compared with the model group, the dehydrodiisobutyronitrile phenol and SASP groups have obviously reduced diarrhea and bloody stool degrees, which shows that both SASP and dehydrodiisobutyronitrile phenol can obviously improve the bloody stool symptoms of mice with ulcerative colitis caused by DSS.

3.1.3 Effect on Colon Length in mice with ulcerative colitis

FIG. 3 shows the effect of dehydrodiisobutyronitrile on colon length in mice with ulcerative colitis. Compared with the normal control group (10.0 +/-0.5 cm), the colon of the model group (5.5 +/-0.5 cm) mice is swollen and bleedings (figure 3A) and is obviously shortened, and the shortening rate is 39.2 percent (figure 3B); the colons of the mice in the SASP group (7.2 ± 0.8cm) and the dehydrodiisobutyronitrile (8.1 ± 0.7cm) were slightly swollen and bleeding (fig. 3A), with shortening rates of 23.1% and 16.6%, respectively (fig. 3B), which were significantly different compared to the model group (P <0.001 and P < 0.05). Therefore, SASP and dehydrodiisobutyronitrile both can obviously improve the colon shortening symptom of mice with the ulcerative colitis caused by DSS.

3.1.4 Effect on Colon histopathology in mice with ulcerative colitis

FIG. 4 shows the effect of dehydrodiisobutyronitrile on repairing colonic tissue damage in mice with ulcerative colitis. Colon tissues of the model group mice are typical inflammatory mucosa manifestations, and can be seen in large-amount inflammatory cell infiltration of mucosa, submucosa and even muscular layer, tissue structure disorder and fracture, submucosa bleeding, edema and telangiectasia; colonic tissue visualization was reduced in the SASP group mice and the dehydrodiisobutyronitrile group mice, ulcers healed, and submucosal bleeding was reduced (fig. 4A).

Normal control mice were scored as 0 on a 1-6 point scale representative of inflammatory tissue injury; the score of the model group mice (5.5 +/-0.3 points) is obviously increased; mice scored less for both the SASP group (4.5 ± 0.4 points) and the dehydrodiisobutyronitrile group (3.5 ± 0.3 points), with significant differences (P <0.001 and P <0.05) compared to the model group (fig. 4B).

The colon histopathological analysis result shows that SASP and dehydrodiisobutyryl phenol can obviously improve the pathological injury of the colon mucosa and inflammatory cell infiltration symptoms of the ulcerative colitis mouse caused by DSS.

The various aspects of the invention are addressed above. It should be understood, however, that equivalent changes and modifications may be made thereto by those skilled in the art without departing from the spirit of the present invention, and that such changes and modifications are intended to be covered by the appended claims.

Claims (1)

1. Application of dehydrodiisobutyronitrile as the only active component in preparing medicine for preventing and treating ulcerative colitis.

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