Pharmaceutical composition containing inulin and fucoidin, pharmaceutical combination and application thereof

Abstract

The invention relates to a medicine composition for treating inflammatory bowel disease, a medicine combination and application thereof. The pharmaceutical composition is prepared from inulin and fucoidin, the medicine combination comprises a first medicine preparation and a second medicine preparation, the active ingredient of the first medicinal preparation is inulin; the active ingredient of the second medicinal preparation is fucoidin. The pharmaceutical composition disclosed by the invention can effectively balance the level of proinflammatory and anti-inflammatory factors in a body, remarkably reduce and control intestinal inflammation, enhance the barrier function of intestinal tracts and relieve oxidative stress injury of tissues, can be used for preventing and treating inflammatory bowel diseases, is high in safety and is suitable for being taken for a long time. The medicine combination can provide self-selection of medicines, is convenient for patients to take medicines, and improves the dependency of the patients.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and in particular to a pharmaceutical composition containing inulin and fucoidan, a drug combination and applications thereof. Background Art

[0002] Inflammatory bowel disease (IBD) is a complex, chronic intestinal disease with a variety of causes, mainly affected by infection, autoimmune response, genetic factors, unhealthy eating habits, mental stress, and side effects of certain drugs. Symptoms of inflammatory bowel disease include diarrhea, bloody stools with mucus, abdominal pain, bloating, etc. It may also lead to serious complications such as heavy blood in the stool, intestinal stenosis, intestinal perforation, and toxic intestinal dilatation, and may even increase the risk of colon cancer. Inflammatory bowel disease not only affects the patient's daily quality of life, but may also cause long-term damage to their physical health, so timely and effective treatment is essential.

[0003] At present, conventional treatments for IBD include drug therapy, surgical treatment, etc. Among them, drug therapy includes the use of anti-inflammatory drugs, immunosuppressants, biological agents, and steroids. These treatments are aimed at controlling inflammation, relieving symptoms, maintaining remission, and improving the patient's quality of life. However, these treatments are often accompanied by side effects.

[0004] Inulin is a mixture of natural fructans, which are fructans formed by fructose molecules connected by β-1,2 bonds. It has physical properties such as high water solubility and low viscosity. In 2001, the American AACC reported that inulin is a soluble dietary fiber that is not decomposed by digestive enzymes in the body. It has multiple physiological functions: promoting the proliferation of bifidobacteria, improving the intestinal microenvironment, controlling blood lipids, stabilizing blood sugar, promoting the synthesis of vitamins and mineral absorption, preventing and treating constipation, and anti-tumor. In 2009, it was approved by my country as a new resource and allowed to be used in food, that is, an active ingredient that can be selectively fermented by beneficial intestinal bacteria or change the composition of beneficial bacteria.

[0005] In recent years, studies have shown that some natural substances such as inulin have the potential to regulate intestinal flora, improve intestinal inflammation and oxidative stress, increase intestinal microbial diversity, regulate intestinal flora, and be used to develop functional foods. Fucoidan is a type of polysaccharide containing L-fucose and sulfate groups, mainly derived from the extracellular matrix of brown seaweed, with antioxidant, anti-inflammatory, antifungal, antiviral and antithrombotic biological activities, and is widely used in nutritional supplements and functional foods.

[0006] The prior art does not disclose inulin and fucoidan as pharmaceutical active ingredients for preventing or treating inflammatory bowel disease. Summary of the invention

[0007] The present invention aims to solve at least one of the technical problems existing in the prior art to a certain extent. To this end, the present invention provides a pharmaceutical composition containing inulin and fucoidan, a drug combination and its use in the preparation of a drug. The pharmaceutical composition of the present invention can effectively balance the level of pro-inflammatory and anti-inflammatory factors in the body, significantly reduce and control intestinal inflammation, enhance intestinal barrier function, alleviate tissue oxidative stress damage, can be used to prevent and treat inflammatory bowel disease, has high safety and is suitable for long-term use.

[0008] In the first aspect of the present invention, the present invention proposes a pharmaceutical composition. According to an embodiment of the present invention, it comprises: inulin and fucoidan. The pharmaceutical composition according to the embodiment of the present invention can effectively balance the levels of pro-inflammatory and anti-inflammatory factors in the body, significantly reduce and control intestinal inflammation, enhance intestinal barrier function, and alleviate tissue oxidative stress damage. It can be used to prevent and treat inflammatory bowel disease, has high safety, and is suitable for long-term use.

[0009] In the second aspect of the present invention, the present invention proposes a drug combination. According to an embodiment of the present invention, it includes: a first drug preparation and a second drug preparation; the active ingredient of the first drug preparation is inulin; the active ingredient of the second drug preparation is fucoidan. The drug combination according to the embodiment of the present invention is the sequential administration of drugs whose active ingredients are inulin and fucoidan, respectively, providing self-selection of medication, facilitating medication for patients, and improving patient dependence.

[0010] In the third aspect of the present invention, the present invention proposes the use of the aforementioned pharmaceutical composition in the preparation of a medicament. According to an embodiment of the present invention, the medicament is used for preventing and / or treating inflammatory bowel disease.

[0011] Those skilled in the art will appreciate that the features and advantages described above for the pharmaceutical composition and drug combination are also applicable to this application and will not be described in detail here.

[0012] In a fourth aspect of the present invention, the present invention provides the use of the aforementioned drug combination in the preparation of a drug. According to an embodiment of the present invention, the drug is used for preventing and / or treating inflammatory bowel disease.

[0013] Those skilled in the art will appreciate that the features and advantages described above for the pharmaceutical composition and drug combination are also applicable to this application and will not be described in detail here.

[0014] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] The above and / or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

[0016] Figure 1 The figures are results of investigation on the effects of inulin and fucoidan intervention on the basic signs of inflammatory bowel disease model mice in Example 1 of the present invention, wherein (A) is a schematic diagram of the modeling of inflammatory bowel disease model mice and the combined drug cycle; (B) is a graph showing the changes in body weight of inflammatory bowel disease model mice under the intervention of inulin and fucoidan; (C) is a graph showing the changes in spleen coefficient of inflammatory bowel disease model mice under the intervention of inulin and fucoidan; (D) is a graph showing the changes in disease activity index (DAI) of inflammatory bowel disease model mice under the intervention of inulin and fucoidan.

[0017] Figure 2 The results of the investigation on the effects of inulin and fucoidan on the colon length and disease activity index of inflammatory bowel disease model mice in Example 1 of the present invention, wherein (A) is a statistical analysis result of the effects of inulin and fucoidan on the colon length of inflammatory bowel disease model mice (n=10; *, p<0.05; **, p<0.01; ***, p<0.001, mean±SEM); (B) is a comparison of the effects of inulin and fucoidan on the colon length of inflammatory bowel disease model mice; (C) is a HE staining image of the colon tissue pathological section of inflammatory bowel disease model mice.

[0018] Figure 3 The results of the investigation on the effects of inulin and fucoidan on the relative expression levels of mRNA of inflammatory factors IL-22, IL-10, IL-17 and IL-8 in the colon of mice with inflammatory bowel disease model in Example 1 of the present invention (n=6; *, p<0.05; **, p<0.01; ***, p<0.001, mean±SEM), wherein (A) is a diagram showing the effects of inulin and fucoidan on the relative expression of IL-22 mRNA in the colon tissue of mice with inflammatory bowel disease model; (B) is a diagram showing the effects of inulin and fucoidan on the relative expression of IL-10 mRNA in the colon tissue of mice with inflammatory bowel disease model; (C) is a diagram showing the effects of inulin and fucoidan on the relative expression of IL-17 mRNA in the colon tissue of mice with inflammatory bowel disease model; (D) is a diagram showing the effects of inulin and fucoidan on the relative expression of IL-8 mRNA in the colon tissue of mice with inflammatory bowel disease model;

[0019] Figure 4The figures are the results of the investigation on the effects of inulin and fucoidan on the relative expression levels of mRNA of inflammatory factors IL-6, IL-1β and TNF-α in the colon of mice with inflammatory bowel disease model in Example 1 of the present invention (n=6; *, p<0.05; **, p<0.01; ***, p<0.001, mean±SEM), wherein (A) is a figure showing the effects of inulin and fucoidan on the relative expression of IL-6 mRNA in the colon tissue of mice with inflammatory bowel disease model; (B) is a figure showing the effects of inulin and fucoidan on the relative expression of IL-1β mRNA in the colon tissue of mice with inflammatory bowel disease model; (C) is a figure showing the effects of inulin and fucoidan on the relative expression of TNF-α mRNA in the colon tissue of mice with inflammatory bowel disease model.

[0020] Figure 5 The figures are the results of the investigation on the effects of inulin and fucoidan on the relative expression levels of mRNA of colon intestinal tight junction proteins in inflammatory bowel disease model mice in Example 1 of the present invention (n=6; *, p<0.05; **, p<0.01; ***, p<0.001, ****, p<0.0001, mean±SEM), wherein (A) is a figure showing the effects of inulin and fucoidan on the mRNA expression levels of Occludin in inflammatory bowel disease model mice; (B) is a figure showing the effects of inulin and fucoidan on the mRNA expression levels of ZO-1 in inflammatory bowel disease model mice; (C) is a figure showing the effects of inulin and fucoidan on the mRNA expression levels of E-cadherin in inflammatory bowel disease model mice; (D) is a figure showing the effects of inulin and fucoidan on the mRNA expression levels of CLDN2 in inflammatory bowel disease model mice; (E) is a figure showing the effects of inulin and fucoidan on the mRNA expression levels of MUC2 in inflammatory bowel disease model mice.

[0021] Figure 6 The figures are the results of the investigation on the effects of inulin and fucoidan on the colon oxidative stress level of IBD model mice in Example 1 of the present invention (n=6; *, p<0.05; **, p<0.01; ***, p<0.001, ****, p<0.0001, mean±SEM), wherein (A) is a figure showing the effects of malondialdehyde (MDA) expression level in colon tissue of IBD model mice; (B) is a figure showing the effects of inulin and fucoidan on superoxide dismutase (SOD) expression level in colon tissue of IBD model mice; (C) is a figure showing the effects of inulin and fucoidan on myeloperoxidase (MPO) expression level in colon tissue of IBD model mice; (D) is a figure showing the effects of inulin and fucoidan on glutathione peroxidase (GSH-Px) expression level in colon tissue of IBD model mice. DETAILED DESCRIPTION

[0022] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be understood as limiting the present invention.

[0023] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. Further, in the description of the present invention, unless otherwise specified, the meaning of "plurality" is two or more.

[0024] The endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0025] In this document, the terms “include” or “comprising” are open expressions, that is, including the contents specified in the present invention but not excluding other contents.

[0026] Pharmaceutical composition

[0027] The present invention proposes a pharmaceutical composition. According to an embodiment of the present invention, the pharmaceutical composition comprises: inulin and fucoidan. The pharmaceutical composition according to an embodiment of the present invention can effectively balance the levels of pro-inflammatory and anti-inflammatory factors in the body, significantly reduce and control intestinal inflammation, enhance intestinal barrier function, and alleviate tissue oxidative stress damage. It can be used to prevent and treat inflammatory bowel disease, has high safety, and is suitable for long-term use.

[0028] According to an embodiment of the present invention, the mass ratio of inulin to fucoidan is (1-3): (3-1), preferably (1-2): (2-1), and more preferably 1: 1. The pharmaceutical composition according to an embodiment of the present invention further improves the effect of preventing or treating inflammatory bowel disease by adjusting the mass ratio of inulin to fucoidan.

[0029] According to an embodiment of the present invention, the pharmaceutical composition comprises 50-200 mg of inulin, 50-200 mg of fucoidan, preferably 100-150 mg of inulin, 100-150 mg of fucoidan. Thus, in order to prevent, alleviate and / or treat inflammatory bowel disease. According to an embodiment of the present invention, the pharmaceutical composition further includes a pharmaceutically acceptable excipient. Such as but not limited to one or more solvents, dispersion media, diluents, dispersing aids, suspension aids, granulation aids, disintegrants, fillers, glidants, liquid vehicles, adhesives, surfactants, isotonic agents, thickeners or emulsifiers, buffers, lubricants, oils, preservatives, flavoring agents, colorants, etc. Excipients such as starch, lactose or dextrin. Pharmaceutically acceptable excipients are well known in the art. Examples of diluents may include, but are not limited to, calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate, lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and / or combinations thereof.

[0030] According to an embodiment of the present invention, the pharmaceutical composition is presented in one of the forms including but not limited to pills, powders, capsules, tablets, granular powders, film-coated agents, orally soluble granules, sachets, dragees or liquids. The pharmaceutical composition of the present disclosure can be prepared by methods well known in the pharmaceutical field.

[0031] In this article, the term "inflammatory bowel disease" is equivalent to "IBD", which is a chronic intestinal inflammatory disease whose cause and pathogenesis are not yet fully understood. It mainly includes two types: ulcerative colitis and Crohn's disease.

[0032] In this article, the term "pharmaceutically acceptable" means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients of the formulation and / or the mammals treated therewith. Preferably, the "pharmaceutically acceptable" of the present invention refers to those approved by federal regulatory agencies or national governments or listed in the U.S. Pharmacopoeia or other generally recognized pharmacopeias for use in animals, particularly humans.

[0033] In this article, the term "pharmaceutically acceptable excipient" may include any solvent suitable for a specific target dosage form. Except for the scope of any conventional excipient being incompatible with the pharmaceutical composition comprising inulin and fucoidan disclosed in the present invention, such as any adverse biological effect produced or interacting with any other component of the pharmaceutically acceptable composition in a harmful manner, their use is also within the scope of the present disclosure.

[0034] Drug combination

[0035] The present invention proposes a drug combination. According to an embodiment of the present invention, it includes: a first drug preparation and a second drug preparation; the active ingredient of the first drug preparation is inulin; the active ingredient of the second drug preparation is fucoidan. The drug combination according to the embodiment of the present invention provides a drug selection for sequential administration of drugs whose active ingredients are inulin and fucoidan, which is convenient for patients to take the drugs and improves the dependence of patients.

[0036] According to an embodiment of the present invention, in the first pharmaceutical preparation and the second pharmaceutical preparation, the mass ratio of inulin to fucoidan is (1-3):(3-1), preferably (1-2):(2-1), and more preferably 1:1.

[0037] According to an embodiment of the present invention, the first pharmaceutical preparation contains 50-400 mg of inulin, and the second pharmaceutical preparation contains 50-400 mg of fucoidan; preferably, the first pharmaceutical preparation contains 100-300 mg of inulin, and the second pharmaceutical preparation contains 100-300 mg of fucoidan. Thus, the purpose of preventing, alleviating and / or treating inflammatory bowel disease and facilitating clinical medication is achieved.

[0038] In some clinical applications of the drug combinations of the present invention, the drug combinations are suitable for combined administration and sequential administration.

[0039] According to an embodiment of the present invention, in the case of combined administration, the drug combination is administered to the subject, and the first drug preparation and the second drug preparation are administered to the subject at the same time for 8 days / cycle, and the dosage is 50 to 200 mg / kg / day of the active ingredient of the first drug preparation and the active ingredient of the second drug preparation, preferably 100 to 150 mg / kg / day.

[0040] According to an embodiment of the present invention, in the case of sequential administration, the drug combination is administered to the subject, with 8 days / cycle, and there are two sequential administration methods: the first sequential administration method is that the first drug preparation is administered to the subject on days 1 to 4, and the dosage is 100 to 400 mg / kg / day, preferably 200 to 300 mg / kg / day, of the active ingredient of the first drug preparation; the second drug preparation is administered to the subject on days 5 to 8, and the dosage is 100 to 400 mg / kg / day, preferably 200 to 300 mg / kg / day, of the active ingredient of the second drug preparation; the second sequential administration method is that the second drug preparation is administered to the subject on days 1 to 4, and the dosage is 100 to 400 mg / kg / day, preferably 200 to 300 mg / kg / day, of the active ingredient of the second drug preparation; the first drug preparation is administered to the subject on days 5 to 8, and the dosage is 100 to 400 mg / kg / day, preferably 200 to 300 mg / kg / day, of the active ingredient of the first drug preparation.

[0041] As used herein, the term "drug combination" refers to the simultaneous or sequential use of two drug preparations for a therapeutic purpose, the result of which is primarily to increase the efficacy of the drugs.

[0042] Pharmaceutical use

[0043] The present invention proposes the use of the aforementioned pharmaceutical composition in preparing a drug. According to an embodiment of the present invention, the drug is used to prevent, alleviate and / or treat inflammatory bowel disease.

[0044] According to an embodiment of the present invention, the inflammatory bowel disease is ulcerative colitis or Crohn's disease.

[0045] Those skilled in the art will appreciate that the features and advantages described above for the pharmaceutical composition and drug combination are also applicable to this use and will not be described in detail here.

[0046] The present invention also proposes the use of the above-mentioned drug combination in the preparation of a drug. According to an embodiment of the present invention, the drug is used to prevent, alleviate and / or treat inflammatory bowel disease.

[0047] According to an embodiment of the present invention, the inflammatory bowel disease is ulcerative colitis or Crohn's disease.

[0048] Those skilled in the art will appreciate that the features and advantages described above for the pharmaceutical composition and drug combination are also applicable to this use and will not be described in detail here.

[0049] Treatment of the disease

[0050] The present invention provides a method for preventing, alleviating and / or treating inflammatory bowel disease. According to an embodiment of the present invention, the method comprises: administering a quantitative amount of the aforementioned pharmaceutical composition or the aforementioned drug combination to a subject.

[0051] According to an embodiment of the present invention, the active ingredients in the pharmaceutical composition are inulin and fucoidan.

[0052] According to an embodiment of the present invention, the drug combination includes: a first drug preparation and a second drug preparation; the first active ingredient of the first drug preparation is inulin; and the second active ingredient of the second drug preparation is fucoidan.

[0053] According to some specific embodiments of the present invention, the method comprises administering the aforementioned pharmaceutical composition to the subject at a rate of 8 days / cycle, wherein inulin and fucoidan are used as active ingredients at a concentration of 50 to 200 mg / kg / day, preferably 100 to 150 mg / kg / day.

[0054] According to some specific embodiments of the present invention, the method comprises administering the aforementioned drug combination to the subject for 8 days / cycle, and administering the first drug preparation and the second drug preparation to the subject at the same time, with the dosage of the active ingredient of the first drug preparation and the active ingredient of the second drug preparation being 50 to 200 mg / kg / day, preferably 100 to 150 mg / kg / day.

[0055] According to some specific embodiments of the present invention, the method comprises administering the aforementioned drug combination to the subject for 8 days / cycle, administering the first drug preparation to the subject on days 1 to 4, at a dosage of 100 to 400 mg / kg / day, preferably 200 to 300 mg / kg / day, of the active ingredient of the first drug preparation; and administering the second drug preparation to the subject on days 5 to 8, at a dosage of 100 to 400 mg / kg / day, preferably 200 to 300 mg / kg / day, of the active ingredient of the second drug preparation.

[0056] According to some specific embodiments of the present invention, the method includes administering the aforementioned drug combination to the subject for 8 days / cycle, administering the second drug preparation to the subject on days 1 to 4, at a dosage of 100 to 400 mg / kg / day, preferably 200 to 300 mg / kg / day, of the active ingredient of the second drug preparation; and administering the first drug preparation to the subject on days 5 to 8, at a dosage of 100 to 400 mg / kg / day, preferably 200 to 300 mg / kg / day, of the active ingredient of the first drug preparation.

[0057] It should be noted that the terms "subject", "individual" and "patient" are used interchangeably herein and refer to a mammal that is evaluated for treatment and / or treated. In one embodiment, the mammal is a human. The terms "subject", "individual" and "patient" include, but are not limited to, individuals with inflammatory bowel disease, such as individuals with ulcerative colitis or Crohn's disease. The subject can be a human, but also includes other mammals, particularly mammals that can be used as laboratory models of human diseases, such as mice, rats, etc.

[0058] In this article, the term "administering" is equivalent to "administering", which refers to introducing a predetermined amount of a substance into a patient by some suitable means. The pharmaceutical composition or drug combination of the present invention can be administered by any common route, as long as it can reach the desired tissue. Various modes of administration are contemplated, including peritoneal, intravenous, intramuscular, subcutaneous injection, etc., but the present invention is not limited to these exemplified modes of administration. Preferably, the pharmaceutical composition or drug combination of the present invention is administered orally.

[0059] As used herein, the term "treatment" refers to the use of drugs to obtain the desired pharmacological and / or physiological effects. The effect may be preventive in terms of completely or partially preventing a disease or its symptoms, and / or may be therapeutic in terms of partially or completely curing a disease and / or the adverse effects caused by the disease. "Treatment" as used herein covers diseases in mammals, particularly humans, and includes: (a) preventing the occurrence of a disease or condition in individuals who are susceptible to the disease but have not yet been diagnosed with the disease; (b) inhibiting the disease, such as blocking the progression of the disease; or (c) alleviating the disease, such as alleviating symptoms associated with the disease. "Treatment" as used herein covers any medication that administers a drug or product to an individual to treat, cure, alleviate, improve, reduce or inhibit an individual's disease, including but not limited to administering a pharmaceutical composition or drug combination as described herein to an individual in need.

[0060] As used herein, the term "prevent" or "preventing" is used to refer to attempting to stop a disease or condition before it occurs.

[0061] The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used are not indicated by the manufacturer and are all conventional products that can be obtained commercially.

[0062] Example 1: Establishment of C57BL / 6 inflammatory bowel disease mouse model and intervention therapy experiment with inulin and fucoidan

[0063] The "C57BL / 6 inflammatory bowel disease mouse model" is an animal model commonly used in laboratory research to simulate human inflammatory bowel disease (IBD). Dextran sulfate sodium (DSS) induces C57BL / 6 mice to exhibit pathological characteristics and clinical symptoms similar to human IBD, which can be used to evaluate new treatments and conduct drug screening.

[0064] The inventors used the C57BL / 6 inflammatory bowel disease (IBD) mouse model to evaluate the therapeutic effects of inulin and fucoidan on IBD. The specific scheme is as follows:

[0065] Fifty 6-week-old C57BL / 6 male mice were randomly divided into a normal group (CON), an inflammatory bowel disease model group (DSS), a fucoidan and inulin co-administration group (Fucoidan+Inulin (1:1)), an inulin-fucoidan administration group (Inulin-Fucoidan), and a fucoidan-inulin administration group (Fucoidan-Inulin), with 10 mice in each group ( Figure 1A). After one week of adaptive feeding, the control group was given normal drinking water, and the other groups of mice were given 2% DSS solution for 14 days. The experimental group was given sequential and combined administration of fucoidan and inulin on the 6th day. The weight of the mice was recorded daily, and the fecal morphology and anal status were observed. After the experiment, the spleen was weighed, the colon length was measured and photographed, and a section of the colon was taken for HE staining to observe intestinal damage. The mouse colon tissue was collected for the determination of the mouse inflammatory factor level, and the remaining samples were stored in a refrigerator at 80°C. Among them, the administration method of the fucoidan and inulin co-administration group (Fucoidan+Inulin (1:1)) was oral administration, the intervention treatment time was eight days, and the dosage was 100 mg / kg / day for fucoidan and inulin respectively; the administration method of the inulin-fucoidan administration group (Inulin-Fucoidan) was oral administration, the intervention treatment time was eight days, inulin was administered for the first four days of the intervention treatment, and fucoidan was administered for the last four days, and the dosage was 200 mg / kg / day; the administration method of the fucoidan-inulin administration group (Fucoidan-Inulin) was oral administration, the intervention treatment time was eight days, inulin was administered for the first four days of the intervention treatment, and fucoidan was administered for the last four days, and the dosage was 200 mg / kg / day.

[0066] The specific investigation methods and results of IBD treatment effects are as follows:

[0067] 1. Disease Activity Index (DAI) score

[0068] During the experiment, the body weight, stool consistency, total rectal bleeding, and survival rate of mice were monitored daily, the occurrence of ulcerative colitis was examined, and the overall disease severity was evaluated by the clinical scoring system (DAI).

[0069] The DAI score is calculated according to Formula 1:

[0070] DAI score = (weight loss score + stool characteristics score + blood in stool score) / 3 Formula 1

[0071] The results are as follows Figure 1 shown.

[0072] 2. Pathological histological changes:

[0073] Hematoxylineosin (HE) staining was used to observe pathological histological changes.

[0074] The specific method is as follows: fix the mouse colon tissue in 4% paraformaldehyde for 24 hours, dehydrate with gradient ethanol, embed in paraffin, make sections, use HE staining, observe its morphological and structural changes under an optical microscope, and observe the pathological changes of colon tissue under an optical microscope after conventional HE staining.

[0075] The results are as follows Figure 2 shown.

[0076] 3. Changes in the expression levels of inflammatory factors and intestinal tight junction proteins

[0077] The transcription levels of inflammatory factors and intestinal tight junction protein mRNA were measured to evaluate the protein expression level. Impaired epithelial barrier function plays an important role in the pathogenesis of colitis. In order to evaluate the protective effect of inulin and fucoidan on the intestinal barrier, the mRNA expression levels of ZO-1, Occludin, E-cadherin, CLDN2 and MUC2 proteins were specifically detected.

[0078] The specific method is: RNA was extracted from the colon tissue near the cecum end of the mouse, and the expression of inflammatory factors such as IL-22, IL-17, IL6, IL8, TNF α, IL1β, IL 10 and proteins such as ZO-1, Occludin, E-cadherin, CLDN2 and MUC2 were analyzed at the transcriptional level by reverse transcription polymerase chain reaction (RTPCR).

[0079] The results are as follows Figure 3 to Figure 5 shown.

[0080] 4. Investigation of oxidative stress level in colonic tissue: Oxidative stress is involved in the onset and development of colitis. To evaluate the effects of inulin and fucoidan on colonic oxidative stress, the contents of MDA, SOD, MPO and tissue GSH-Px activity in colonic tissue were determined. The kits of Shanghai Sangon Biotech Co., Ltd. were used to detect the levels of superoxide dismutase (SOD), malondialdehyde (MDA), myeloperoxidase (MPO) and glutathione peroxidase (GSH-Px) in colonic tissue.

[0081] The results are as follows Figure 6 shown.

[0082] The results show:

[0083] (1) After 14 days of DSS administration, the body weight of mice in the DSS model group decreased significantly; compared with the CON group, the colon length of mice in the DSS model group was significantly reduced ( Figure 2 A. Figure 2 B), and the DAI score increased significantly ( Figure 1 D); The colon tissue in the DSS model group was severely damaged, with symptoms such as loss of crypt structure and inflammatory cell infiltration ( Figure 2C). As a result, the mice in the DSS group showed pathological characteristics and clinical symptoms similar to those of human IBD, and the C57BL / 6 inflammatory bowel disease mouse model was successfully established, which can be used to evaluate the therapeutic effects of inulin and fucoidan on IBD.

[0084] (2) Compared with the DSS model group, administration of inulin and fucoidan suppressed the symptoms of colon shortening and spleen enlargement in mice with inflammatory bowel disease ( Figure 1 C), and the DAI score was significantly reduced; after the administration of inulin and fucoidan, the colon damage was significantly reduced, the colon tissue structure and mucosal epithelial cells were significantly improved, the infiltration of inflammatory cells was significantly reduced, and the normal physiological morphology of most cells was maintained. This shows that intestinal inflammation was alleviated and effectively controlled, and the treatment effect of inulin and fucoidan was good.

[0085] (3) Compared with the CON group, the mRNA transcription levels of inflammatory factors IL-17, IL6, TNFα, and IL1β in the colon tissue of mice in the DSS group were increased, while the mRNA transcription level of the anti-inflammatory factor IL10 was decreased. Compared with the DSS model group, the mRNA transcription levels of TNFα and IL-8 in the colon tissue of the inulin and fucoidan (1:1) administration group were significantly downregulated, and the expression level of IL-22 was significantly upregulated. In the inulin-fucoidan administration group, the expression level of IL-6 was significantly downregulated, and the expression level of IL-10 was significantly increased. The level of the pro-inflammatory cytokine IL1β in the fucoidan-inulin administration group was significantly decreased ( Figure 3 , Figure 4 ). It can be seen that the combined administration or sequential administration of inulin and fucoidan has a significant effect on improving the balance of pro-inflammatory and anti-inflammatory factors in mice with ulcerative colitis.

[0086] (4) In the DSS model group, DSS intervention destroyed the intestinal epithelial barrier of mice and significantly reduced the mRNA expression levels of intestinal occludin and tight junction proteins ( Figure 5 A. Figure 5 B. Figure 5 C. Figure 5 D. Figure 5 E). Compared with the DSS model group, the inulin-fucoidan group significantly increased the expression of ZO1, Occludin, E-cadherin and MUC2 proteins in colon tissue, and the fucoidan-inulin group significantly increased the mRNA expression level of CLDN2 protein.

[0087] (5) Compared with the CON group, the MDA content in the colon tissue of the DSS model group increased significantly, and the inulin and fucoidan (1:1) administration group and the fucoidan-inulin administration group significantly reversed the trend of MDA content increase ( Figure 6A); Compared with the CON group, the colonic MPO level in the DSS model group increased significantly, and the activities of SOD and GSH-Px decreased significantly, while the inulin and fucoidan (1:1) co-administration group, inulin-fucoidan administration group, and fucoidan-inulin administration group all significantly reversed this trend, especially the trend of decreased SOD content and decreased GSH-Px content ( Figure 6 B. Figure 6 D). It can be seen that the combined administration or sequential administration of inulin and fucoidan effectively alleviates the oxidative stress damage in the colon tissue of colitis mice in vivo.

[0088] The above results show that a pharmaceutical composition comprising inulin and fucoidan or a drug combination in which the active ingredients are inulin and fucoidan respectively can effectively prevent or treat inflammatory bowel disease. Compared with existing drugs or surgical treatments, it has high safety, is suitable for long-term use, can meet different clinical medication needs, and has high drug development value.

[0089] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A pharmaceutical composition, characterized in that include: Inulin and fucoidan.

2. The pharmaceutical composition according to claim 1, characterized in that The mass ratio of inulin to fucoidan is (1-3):(3-1), preferably (1-2):(2-1), and more preferably 1:

1.

3. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition comprises 50-200 mg of inulin, 50-200 mg of fucoidan, preferably 100-150 mg of inulin, and 100-150 mg of fucoidan; Optionally, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

4. A drug combination, characterized in that include: a first pharmaceutical preparation and a second pharmaceutical preparation; The active ingredient of the first pharmaceutical preparation is inulin; The active ingredient of the second pharmaceutical preparation is fucoidan.

5. The drug combination according to claim 4, characterized in that: In the first and second pharmaceutical preparations, the mass ratio of inulin to fucoidan is (1-3):(3-1), preferably (1-2):(2-1), and more preferably 1:

1.

6. The drug combination according to claim 4, characterized in that: The first pharmaceutical preparation contains 50-400 mg of inulin, and the second pharmaceutical preparation contains 50-400 mg of fucoidan; preferably, the first pharmaceutical preparation contains 50-400 mg of inulin, and the second pharmaceutical preparation contains 100-300 mg of fucoidan.

7. Use of the pharmaceutical composition according to any one of claims 1 to 3 in the preparation of a drug, characterized in that: The medicament is used for preventing and / or treating inflammatory bowel disease.

8. The use according to claim 7, characterized in that The inflammatory bowel disease is ulcerative colitis or Crohn's disease.

9. Use of the drug combination according to claims 4 to 6 in the preparation of a drug, characterized in that: The medicament is used for preventing and / or treating inflammatory bowel disease.

10. The use according to claim 9, characterized in that The inflammatory bowel disease is ulcerative colitis or Crohn's disease.

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