Application of glaucocalyxin A in preparation of medicine for preventing or treating colitis

Abstract

The invention discloses application of glaucocalyxin A in preparation of a medicine for preventing or treating colitis, and particularly relates to application of glaucocalyxin A in preparation of a medicine for preventing or treating ulcerative colitis. A mouse ulcerative colitis model is established through induction, glaucocalyxin A is adopted as a treatment medicine, and the DAI score, the colon length, the spleen index, the intestinal tissue structure, the inflammatory factor level, the LPS level and changes of MUC2, ZO-1 and Occludin protein expression in colon tissue before and after treatment of the mouse ulcerative colitis model are observed; the treatment effect and the action mechanism of the glaucocalyxin A on the ulcerative colitis are elaborated, so that a scientific basis is provided for preventing or treating the ulcerative colitis by the glaucocalyxin A.

Description

Technical Field

[0001] The invention relates to the field of biomedicine, and in particular to application of bluecalyx acetone in preparing medicines for preventing or treating colitis. Background Art

[0002] Ulcerative colitis (UC) is a nonspecific inflammatory disease of unknown cause. The lesions are located in the large intestine and are distributed continuously and diffusely, mostly involving the rectum and sigmoid colon. Its typical clinical course includes bloody stools with mucus, pus, and abdominal pain, diarrhea, a long course of disease, and is prone to recurrence and even cancer, which seriously affects the patient's quality of life. At present, the true cause of UC is still unclear. Genetic factors, smoking, diet, living environment, infection, immunity, mental and psychological factors may promote the occurrence and development of this disease. Clinically, the treatment of UC is mainly based on drugs, mainly including: (1) aminosalicylic acid preparations, such as sulfasalazine, which have a long medication cycle and are prone to dose-related adverse reactions, such as gastrointestinal symptoms, urticaria, dyspnea, joint pain, liver toxicity, etc.; (2) Glucocorticoids and immunosuppressants have many adverse reactions and cannot control the recurrence of UC; (3) Anti-tumor necrosis factor α monoclonal antibodies are prone to recurrence and have many adverse reactions; (4) Probiotic therapy improves UC symptoms The effect is general and it is often used in combination with other types of drugs in clinical practice; (5) Enema treatment with tin powder, gentamicin, 2% procaine and dexamethasone has poor patient compliance (Li Nan, Research Progress in Ulcerative Colitis, Jilin Medicine, 2014, 35(7): 1486-1487.; Ma Qinglin, Du Lidong, Zang Kaihong, Sun Min, Cao Rubing, Ren Yuan, Overview of Research Progress in Ulcerative Colitis, Journal of Medical Forum, 2020, 41(1): 175-177. Taku Kobayashi, Britta Siegmund, Catherine Le Berre, Shu Chen Wei, Marc Ferrante, Bo Shen, Charles N. Bernstein, Silvio Danese, Laurent Peyrin-Biroulet & Toshifumi Hibi. Ulcerative colitis. Nature Reviews Disease Primers. 2020, 6: 74.).

[0003] The plant of the genus Glaucae is widely distributed in Northeast China, North China, Shandong, Jiangsu, Anhui, Henan and other provinces. Among them, glaucocalyxin A (GLA) is the main component of Glaucae, with a content of more than 1.0% in the plant. It has sufficient sources, stable preparation process, high safety, and has been found to have certain anti-tumor and anti-liver fibrosis activities. At present, there is no report on whether glaucocalyxin A has the effect of treating colitis. Summary of the invention

[0004] To solve the above problems, the present invention provides the use of bluecalyx A in the preparation of a drug for preventing or treating colitis, and verifies the therapeutic effect and mechanism of bluecalyx A on ulcerative colitis through animal experiments, providing a scientific basis for the use of bluecalyx A in preventing or treating ulcerative colitis.

[0005] Specifically, the present invention provides the following technical solutions:

[0006] The first aspect of the present invention provides the use of bluecalyx A in preparing a medicine for preventing or treating colitis.

[0007] Furthermore, the colitis is preferably ulcerative colitis.

[0008] Furthermore, the bluecalyx factor A is used to reduce the levels of inflammatory factors such as TNF-α, IL-1β, IL-6, IL-17 and IL-23 in serum.

[0009] Furthermore, the bluecalyx factor A is used to reduce the level of lipopolysaccharide (LPS) in serum.

[0010] The intestinal mucosal barrier is of great significance for maintaining intestinal integrity. The destruction of the intestinal mucosal barrier will increase the contact between host cells and bacteria and toxins in the intestine. The pathological invasion and colonization of bacteria will further stimulate the body's immune response and the secretion of multiple cytokines, promote the occurrence and development of ulcerative colitis and the development of inflammation. The reduction of the mucus layer is also one of the most common pathological features of patients with ulcerative colitis. The mucus layer is located above the intestinal epithelial layer and is mainly composed of mucin and mucopolysaccharides secreted by goblet cells, of which mucin accounts for the majority, and mucin is mainly composed of MUC2 protein secreted by goblet cells.

[0011] Furthermore, the bluecalyx acetone can be used to increase the expression of MUC2 protein in colon tissue, thereby protecting the intestinal mucosal layer.

[0012] Intestinal epithelial cells are connected by tight junction proteins, which prevent ions, small molecules and bacteria from passing freely. Intercellular tight junction proteins are essential for maintaining the permeability of the intestinal epithelial layer and play a decisive role in the intestinal mucosal barrier function. Among them, Occludin protein has the function of regulating intercellular permeability. ZO-1 protein mainly regulates intercellular material transport and the polarity of epithelial cells. Upregulating the expression of tight junction proteins helps maintain the structural integrity of the intestinal mucosal barrier, thereby preventing the occurrence of ulcerative colitis.

[0013] Furthermore, the bluecalyx factor A can be used to increase the expression of ZO-1 protein and Occludin protein in colon tissue, thereby protecting the tight junctions between intestinal epithelial cells and facilitating the maintenance of epithelial barrier integrity.

[0014] The second aspect of the present invention provides a drug for preventing and treating ulcerative colitis, wherein the active ingredient of the drug comprises cyanocalyxin A.

[0015] Furthermore, the drug is preferably administered via the gastrointestinal tract.

[0016] Furthermore, the drug also contains one or more pharmaceutically acceptable excipients or carriers.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] 1. The present invention applies blue calyx A to the prevention and treatment of ulcerative colitis for the first time, and conducts animal experimental research on the prevention and treatment of ulcerative colitis by constructing a mouse ulcerative colitis model. The study finds that blue calyx A has a protective effect on ulcerative colitis, and finds that the mechanism of action of preventing and treating ulcerative colitis is related to its significant reduction of TNF-α, IL-1β, IL-6, IL-17, IL-23, LPS levels in serum and the increase of MUC2 protein, Occludin protein and ZO-1 protein expression in colon tissue. Under the effect of blue calyx A, the DAI score of mice is significantly reduced, and the symptoms of colon congestion, redness and swelling and splenomegaly are alleviated, and the intestinal tissue structure is normal, and no obvious inflammatory cell infiltration is seen. It can be seen that blue calyx A can effectively prevent and treat ulcerative colitis.

[0019] 2. The present invention studies the therapeutic effect and mechanism of bluecalyx A on ulcerative colitis by establishing an animal model, providing a scientific basis for the use of bluecalyx A in preventing or treating ulcerative colitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The figure shows the changes in weight and DAI scores of mice in different experimental groups, compared with the blank group. ## P<0.01, ### P < 0.001,#### P<0.0001; compared with the model group, * P<0.05, ** P<0.01, *** P < 0.001, **** P < 0.0001;

[0021] Figure 2 The colon of mice in different experimental groups;

[0022] Figure 3 The changes of colon length and spleen index of mice in different experimental groups are compared with the blank group. ### P < 0.001, #### P<0.0001; compared with the model group, ** P<0.01, *** P < 0.001;

[0023] Figure 4 The sections of the colon tissues of mice in different experimental groups were stained (H&E, 200×), a is the blank group, b is the model group, and c is the blue calycin group;

[0024] Figure 5 Serum inflammatory factors and LPS levels of mice in different experimental groups were compared with those in the blank group. #### P<0.0001; compared with the model group, **** P < 0.0001;

[0025] Figure 6 Immunofluorescence images of ZO-1, Occludin, and MUC2 proteins in the colon tissues of mice in different experimental groups;

[0026] Figure 7 Comparison of ZO-1, Occludin, and MUC2 protein levels in colon tissues of mice in different experimental groups, compared with the blank group. # P<0.05, ## P<0.01, compared with the model group, ** P<0.01, *** P<0.001. DETAILED DESCRIPTION

[0027] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it, but the embodiments are not intended to limit the present invention.

[0028] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0029] Example

[0030] This example studies the effect of blue calyx A on the pharmacological indicators of ulcerative colitis mice, including weight change, DAI score, spleen index, colon length, colon tissue, serum inflammatory factor level, serum LPS level and protein expression in colon tissue. The specific operation is as follows:

[0031] 1. Experimental medicinal materials and reagents

[0032] Preparation of blue calyx amine: 2.0 kg of blue calyx amine leaf and stem were cut into sections, and then refluxed and extracted with 3.0 L of 80% ethanol for 1.5 h, and extracted twice. The filtered extract was recovered by decompression to obtain an extract. The extract was dissolved in water, extracted with petroleum ether and chloroform in turn, and the chloroform layer was recovered to obtain 24 g of chloroform extract, which was then chromatographed on an 800 g silica gel column with elution solvents of chloroform: methanol (100:1, 50:2) to obtain component 1 and component 2 (2.2 grams). From component 2, 0.1 g of colorless needle-like blue calyx amine was obtained by crystallization with methanol. The characterization results of the product are shown below:

[0033] MP: 220.0-222.0℃;

[0034] EIMS m / z:355.2[M+Na] + ;

[0035] 1 H NMR(400MHz,DMSO-d6)δ:5.95(1H,brs,17-Hb), 5.40(1H,brs,17-Ha),4.71(1H,brs,H- 14α),4.03(1H,m,7β-H),2.95(1H,brs,13α-H),1.04(3H,s),1.00(3H,s),0.98(3H,s);

[0036] 13C NMR(100MHz,DMSO-d6)δ:37.9(1-C),33.9(2-C),216.5(3-C),46.5(4-C),50.6(5-C),30.9(6-C),73.0(7-C),60.7(8-C),52.8(9-C),38.6(1 0-C),18.1(11-C),30.3(12-C),46.1(13-C),74.6(14-C),207.0(15-C ),148.8(16-C),116.9(17-C),27.4(18-C),21.1(19-C),18.1(20-C);

[0037] The purity of the prepared bluecalyx A by HPLC area normalization method was over 98%; HPLC instrument: Aglient Infinitely 1260, chromatographic column: YMC-Pack ODS-A (5μm, 250×4.6mm); column temperature: 25°C, mobile phase: methanol: water (60:40), bluecalyx A 0.997 mg / mL.

[0038] Dextran sulfate sodium was purchased from Shanghai Xibao Biotechnology Co., Ltd.; 4% paraformaldehyde solution was purchased from Bio-Time; hematoxylin-eosin stain was purchased from Jiangsu Addison Biotechnology Co., Ltd.; toluene, ammonia water, hydrochloric acid, anhydrous ethanol and neutral gum were purchased from Sinopharm Chemical Reagent Co., Ltd.; sulfasalazine enteric-coated tablets were purchased from Shanghai Xinyi Tianping Pharmaceutical Co., Ltd.; ELISA kits for TNF-α, IL-1β, IL-6, IL-17, IL-23 and LPS were purchased from Jiangsu Sumeike Biotechnology Co., Ltd.; anti-ZO-1 rabbit antibody and anti-occludin rabbit antibody were purchased from Wuhan Sanying Biotechnology Co., Ltd.; anti-MUC2 rabbit antibody was purchased from Google Biotechnology Co., Ltd.; HRP-labeled goat anti-rabbit was purchased from SeraCare, and other chemical reagents were of analytical grade.

[0039] 2. Experimental instruments: 702-80℃ refrigerator, produced by Thermo Fisher Scientific Inc., USA; TECAN full-wavelength microplate reader M1000 pro, produced by Nanjing Kelinde Scientific Instrument Co., Ltd.; incubator, produced by Shanghai Jinghong Experimental Equipment Co., Ltd.; balance, produced by Ohaus Instrument Co., Ltd.; water bath, produced by Shanghai Ailan Instrument Co., Ltd.; pathological slicer, produced by Shanghai Leica Instrument Co., Ltd.; tissue spreader, produced by Cody Instrument Equipment Co., Ltd., Jinhua City, Zhejiang Province; oven, produced by Shanghai Huitai Instrument Manufacturing Co., Ltd.; slides and cover slips, produced by Jiangsu Shitai Experimental Equipment Co., Ltd.); inverted optical microscope and imaging system, produced by Nikon, Japan; dehydrator, embedding machine and freezing table, produced by Wuhan Junjie Electronics Co., Ltd.

[0040] 3. Experimental methods

[0041] Drug preparation: Weigh an appropriate amount of blue calyx methyl cellulose and dissolve it in 0.5% sodium carboxymethyl cellulose to make 60 mg / kg; dissolve dextran sulfate sodium (DSS) in deionized water to make a 2.5% DSS aqueous solution, and replace it every 2 days.

[0042] Animal grouping, modeling and drug administration: After adaptive culture, male C57BL / 6J mice were divided into blank group (n=5), DSS model group (n=5), and DSS+coleucine group (n=5). Except for the blank group, which was given deionized water, the other groups were allowed to drink DSS aqueous solution freely to induce the mouse ulcerative colitis (UC) model for 7 consecutive days, and deionized water was replaced from the 8th day. In the DSS+coleucine group, oral administration began on the second day of modeling, once a day for 8 consecutive days, and the administration was ensured to be at the same time every day; the blank group and the model group were given normal saline by oral administration.

[0043] 3.1. Study the effect of cyanocalyxin A on body weight change and DAI index score of UC mice:

[0044] During the experiment, the general condition, eating, drinking, weight change, stool characteristics, fecal occult blood / visual blood in each group of mice were observed and recorded in detail every day. Starting from the 0th day of the experiment, the weight of each group of mice was weighed at a fixed time every day, the fecal characteristics of each mouse were recorded, and the fecal occult blood was detected using a fecal occult blood qualitative detection kit (o-tolidine method). The change in body weight was expressed as a percentage of the baseline body weight decrease: body weight change (%) = [1 + (body weight after modeling - body weight before modeling) / body weight before modeling] × 100%.

[0045] According to the Cooper scoring system (as shown in Table 1 below), the disease activity index (DAI) of the colon tissue of mice was assessed daily based on the weight loss, stool characteristics, and bloody stools of the mice. Each index of the score was scored on a scale of 0-4, and DAI = (weight loss score + stool characteristics score + bloody stool score) / 3.

[0046] Table 1 Disease Activity Index (DAI) Scoring Criteria

[0047] Weight loss / % Blood in stool Stool characteristics Rating value / points ＜1 Negative normal 0 ≥1-5 - - 1 ≥5-10 Positive, but no visible blood in the stool Thin and soft 2 ≥10-15 - - 3 ≥15 Visible blood in the stool Watery stools 4

[0048] The results are as follows: During the above experiment, the mice in the blank group were in good mental state, sensitive, had a relatively stable diet, slow weight gain, shiny fur, and normal stool. The mice in the model group showed symptoms such as listlessness, curling and arching the back, and mucus and blood in the stool. Figure 1 The figure shows the weight changes and DAI scores of mice in different experimental groups. It can be seen from the figure that the weight of mice in the model group showed a downward trend, and the weight loss was very significant from the 5th day (P<0.0001); blue calyx A can reduce the weight loss of UC mice, especially on the 7th day, blue calyx A significantly reduced the weight loss of mice (P<0.0001). Compared with the blank group, the DAI score of mice in the model group increased significantly (P<0.0001); compared with the model group, the DAI score of mice in the blue calyx A group (P<0.0001) was significantly reduced.

[0049] 3.2. Study the effect of cyanocalyxin A on colon length and spleen index of UC mice:

[0050] Blood was collected 1 hour after the last administration, and after standing for 1 hour, the supernatant was collected by centrifugation at 4°C, 3500r / min for 10 minutes. After the mice were killed, the distance from the ascending colon to the end of the rectum was measured as the length of the mouse colon; the colon segment was divided into two parts, one of which was immediately placed in a -80°C refrigerator; the other was fixed in a fixative. After the spleen was separated by an instrument, the surface was rinsed with saline, the surface moisture was absorbed with filter paper, and the weight was weighed to calculate the spleen index, spleen index = spleen weight (mg) / mouse body weight (g).

[0051] The results are as follows: Figure 2 , 3 It can be seen that compared with the blank group, the colon of the mice in the model group was significantly congested and the length was significantly shortened (P<0.0001), the spleen of the mice in the model group was enlarged, and the spleen index was significantly increased (P<0.001); compared with the model group, the colon congestion symptoms and colon length of the mice in the bluecalyx A group were significantly improved (P<0.001), the spleen enlargement was alleviated, and the spleen index was significantly reduced (P<0.01).

[0052] 3.3. Study the effect of cyanocalyxin A on colon tissue of UC mice:

[0053] Take out the colon tissue of the mouse from the fixative, flatten the tissue and put it into the dehydration box, dehydrate, wax and embed in the dehydrator in gradient alcohol. Put it into the -20℃ freezer to cool, take it out after the wax solidifies, and trim the wax block. Slice it on the slicer, the slice thickness is 4μm, put it on the 40℃ warm water of the spreader, flatten the tissue, bake it in a 60℃ oven, and store it at room temperature for use. According to the operation of the staining kit, dewax, stain, dehydrate, seal with neutral gum, and finally observe the pathological changes of each tissue under an optical microscope.

[0054] The results are as follows: Figure 4 The following are the sections of the colon tissue of mice in different experimental groups (H&E, 200×). As can be seen from the figure, the intestinal tissue structure of the blank group (Figure a) mice was normal, the epithelial cells of the mucosal layer were arranged neatly and tightly in the field of view, and no shedding was observed; the crypt structure was intact and arranged neatly; no edema was observed in the submucosal layer; and no obvious inflammatory cell infiltration was observed in the tissue. Compared with the blank group, the intestinal tissue structure of the UC mice (Figure b) was severely abnormal, the epithelial cells of the mucosal layer were shedding in the field of view, and the lamina propria was exposed, as shown by the red arrow; the crypt structure disappeared, as shown by the blue arrow; no edema was observed in the submucosal layer; and a large number of inflammatory cell infiltrations were observed in the tissue, as shown by the yellow arrow. Compared with the model group, the intestinal tissue structure of the mice in the blue calyx A group (Figure c) was normal, the epithelial cells of the mucosal layer were arranged neatly and tightly in the field of view, and no shedding was observed; the crypt structure was intact and arranged neatly; no edema was observed in the submucosal layer; and no obvious inflammatory cell infiltration was observed in the tissue.

[0055] 3.4. Study the effect of cyanocalyxin A on the levels of serum inflammatory factors and serum LPS in UC mice:

[0056] One hour after the last administration, 10% chloral hydrate was injected intraperitoneally for anesthesia, and blood was collected from the abdominal aorta with a blood collection needle for analysis of biochemical indicators in the blood.

[0057] The results are as follows: Figure 5 The figures show the levels of serum inflammatory factors and serum LPS in mice in different experimental groups. As shown in the figure, the levels of serum TNF-α, IL-1β, IL-6, IL-17, IL-23 and LPS in the model group were significantly higher than those in the blank group (P<0.0001); compared with the model group, the levels of serum TNF-α, IL-1β, IL-6, IL-17, IL-23 and LPS in the bluecalyx A group were significantly reduced (P<0.0001). It can be seen that the levels of TNF-α, IL-1β, IL-6, IL-17, IL-23 and LPS in the serum of DSS-induced UC mice were significantly increased. After intervention with bluecalyx A, the levels of various inflammatory factors and LPS were significantly reduced, which also shows that bluecalyx A can reduce the levels of serum TNF-α, IL-1β, IL-6, IL-17, IL-23 and LPS in UC mice to play a therapeutic role in ulcerative colitis in mice.

[0058] 3.5. Study the effect of cyanocalyxin A on the expression of MUC2, ZO-1 and Occludin proteins in colon tissue of UC mice:

[0059] The sections of the above colon tissues were dewaxed, antigen repaired, blocked, and serum blocked. Then, primary antibodies (anti-ZO-1 rabbit antibody 1:200, anti-occludin rabbit antibody 1:500, and anti-MUC2 rabbit antibody 1:1000) were added and incubated in a humidified box overnight at 4°C. The slides were placed in PBS (pH 7.4) and washed on a decolorizing shaker for 3 times, each time for 5 minutes. After the sections were slightly dried, the secondary antibody (HRP-labeled goat anti-rabbit 1:400) was added dropwise in the circle to cover the tissue, incubated at room temperature in the dark for 50 minutes, TSA reagent was added and incubated at room temperature for 20 minutes, DAPI was used to counterstain the cell nuclei and incubated at room temperature in the dark for 10 minutes, the sections were sealed, and the sections were observed under a fluorescence microscope and image analysis was collected.

[0060] The results are as follows: Figure 6 , 7 The immunofluorescence images and content comparison images of ZO-1, Occludin and MUC2 proteins in the colon tissues of mice in different experimental groups are shown in Figure 1. Compared with the blank group, the expression of MUC2, ZO-1 and Occludin proteins in the colon tissues of mice in the model group was significantly reduced (P<0.05, P<0.01, P<0.01); compared with the model group, the expression of MUC2, ZO-1 and Occludin proteins in the colon tissues of mice in the blue calyx A group was significantly increased (P<0.01, P<0.01, P<0.001), and exceeded the blank group. It can be seen that the expression of MUC2, ZO-1 and Occludin proteins in the colon tissues of DSS-induced UC mice was significantly downregulated, and after intervention with blue calyx A, the expression of the three proteins was significantly upregulated, which also shows that blue calyx A can protect UC mice by protecting the intestinal mucosal layer and protecting the tight junctions between intestinal epithelial cells.

[0061] In summary, blue calyx A can effectively reduce the DAI score of ulcerative colitis mice, reduce the degree of colon shortening, alleviate the symptoms of colon congestion and swelling, relieve splenomegaly, and reduce spleen index; and further studies have found that blue calyx A can effectively reduce the levels of TNF-α, IL-1β, IL-6, IL-17, IL-23 and LPS in the serum of ulcerative colitis mice, improve the level of inflammation in ulcerative colitis mice, and effectively increase the expression of MUC2 protein, Occludin protein and ZO-1 protein in the colon tissue of ulcerative colitis mice, thereby protecting the tight junctions between intestinal epithelial cells and the intestinal mucosal layer, thereby protecting the integrity of the intestinal mucosal barrier and improving colon tissue damage. The present invention verifies the therapeutic effect and mechanism of blue calyx A on ulcerative colitis through the above-mentioned animal experiments, and provides a scientific basis for the use of blue calyx A in the prevention or treatment of ulcerative colitis.

[0062] The above-described embodiments are only preferred embodiments for fully illustrating the present invention, and the protection scope of the present invention is not limited thereto. Equivalent substitutions or changes made by those skilled in the art based on the present invention are within the protection scope of the present invention. The protection scope of the present invention shall be subject to the claims.

Claims

1. Application of bluecalyx acetone in the preparation of medicines for preventing or treating colitis.

2. The use according to claim 1, characterized in that: The colitis is ulcerative colitis.

3. The use according to claim 1 or 2, characterized in that: The bluecalyx acetone is used to reduce the levels of inflammatory factors such as TNF-α, IL-1β, IL-6, IL-17 and IL-23 in serum.

4. The use according to claim 1 or 2, characterized in that: The bluecalyx acetone is used for reducing the level of lipopolysaccharide in serum.

5. The use according to claim 1 or 2, characterized in that: The bluecalyx acetone is used for increasing the expression of MUC2 protein in colon tissue.

6. The use according to claim 1 or 2, characterized in that: The bluecalyx acetone is used for increasing the expression of ZO-1 protein in colon tissue.

7. The use according to claim 1 or 2, characterized in that: The bluecalycin A is used for increasing the expression of Occludin protein in colon tissue.

8. A drug for preventing and treating ulcerative colitis, characterized in that: The active ingredient of the medicine comprises bluecalyx A.

9. The drug according to claim 8, characterized in that The drug is administered via the gastrointestinal tract.

10. The drug according to claim 8, characterized in that The drug further comprises one or more pharmaceutically acceptable excipients or carriers.

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