JP2000309537A - Prevention and/or curing agent for entric disease - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject prevention and/or curing agent having excellent effect for preventive and/or curing enteric disease, especially an enteral hemorrhage due to inflammatory enteric disease by including glycosaminoglycan sulfate. SOLUTION: This agent contains glycosaminoglycan sulfate, preferably chondroitin sulfate having 10,0000-40,000 molecular weight or its salt (e.g. sodium chondroitin sulfate) as an active component. The glycosaminoglycan sulfate, e.g. chondroitin sulfate in a shape of proteoglycan is obtained by e.g. using a gristle of an aquatic animal such as a shark, a mammal such as cattle or swine or birds as a raw material, extracting by a known method such as a neutral salt method, removing fat.solid component, etc., and drying. Preferably, an administration amount of the agent is daily 20-500 mg as a weight of glycosaminoglycan sulfate of the active component per 1 kg of a body weight. Preferably, 0.1-90 wt.% of glycosaminoglycan is contained in the objective formulation.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a prophylactic and / or therapeutic agent for intestinal diseases containing a sulfated glycosaminoglycan or a pharmaceutically acceptable salt thereof.

[0002]

2. Description of the Related Art Sulfated glycosaminoglycans such as chondroitin sulfate are a group of glycosaminoglycans having a sulfate group among glycosaminoglycans generally distributed in connective tissues of animals, mainly cartilage. It binds to proteins and exists as proteoglycans.

[0003] Sulfated glycosaminoglycans, especially chondroitin sulfate, are usually used as sodium salts,
Pharmacologically, it is known to have a fat serum clarifying action, an anticoagulant action, a cell activating action, a detoxifying action, an analgesic action and the like.

[0004] Sulfated glycosaminoglycan is a safe biological substance. In addition to the various pharmacological actions known so far as described above, if a new pharmacological action can be found, side effects will occur. It is expected to be used as a small number of new medicines.

[0005]

SUMMARY OF THE INVENTION Accordingly, an object of the present invention is to find a new pharmacological action of sulfated glycosaminoglycan, and to provide a medicine utilizing the pharmacological action.

[0006]

The present inventors have conducted intensive studies on the pharmacological action of sulfated glycosaminoglycans. As a result, sulfated glycosaminoglycans such as chondroitin sulfate have been developed. It has been found that it has a preventive and therapeutic effect on bowel diseases, especially inflammatory bowel diseases, and has an effect of remarkably preventing and reducing intestinal bleeding due to inflammatory bowel diseases. The present invention has been completed based on such findings.

[0007] Accordingly, the present invention provides a preventive and / or therapeutic agent for intestinal disease containing a sulfated glycosaminoglycan or a pharmaceutically acceptable salt thereof.

According to a preferred embodiment of the agent for preventing and / or treating intestinal diseases according to the present invention, the sulfated and / or glycosaminoglycan is derived from sharks and / or bovines. The above-mentioned intestinal disease preventive and / or therapeutic agent wherein the intestinal disease is an inflammatory bowel disease; the above-mentioned intestinal disease preventive and / or therapeutic agent wherein the inflammatory bowel disease is an inflammatory bowel disease accompanied by bleeding; Wherein the inflammatory bowel disease associated with bleeding is ulcerative colitis;
Alternatively, a therapeutic agent is provided.

[0009]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail.

The agent for preventing and / or treating intestinal diseases of the present invention is characterized by containing one or more sulfated glycosaminoglycans or pharmacologically acceptable salts thereof as an active ingredient.

[0011] The sulfated glycosaminoglycan used in the present invention comprises an amino sugar and uronic acid (or galactose).
Among glycosaminoglycans having a repeating structure of a disaccharide unit consisting of: a compound having a sulfate group and a compound widely distributed in connective tissues such as cartilage of animals.

Specific examples of sulfated glycosaminoglycans include chondroitin sulfate (chondroitin sulfate A, chondroitin sulfate C, chondroitin sulfate E, chondroitin sulfate K, chondroitin polysulfate, etc.), dermatan sulfate (chondroitin sulfate B), keratan sulfate (Including keratan polysulfate), heparan sulfate, sulfated hyaluronic acid, and the like, but are not limited thereto.

[0013] Among these sulfated glycosaminoglycans, chondroitin sulfate is particularly preferred. Chondroitin sulfate generally has a molecular weight of several thousand to about 50,000,
It is preferably about 10,000 to 40,000, particularly preferably 20,000 to 30,000.

The sulfated glycosaminoglycan used in the present invention may be a commonly used pharmacologically acceptable salt. Pharmaceutically acceptable salts of sulfated glycosaminoglycans include, for example, alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt and magnesium salt; cyclohexylamine, trialkylamine, Salts with organic bases such as pyridine and amino acids; and salts with inorganic bases such as ammonium salts, etc., are not particularly limited, but alkali metal salts are preferable.
Sodium salts are particularly preferred.

The sulfated glycosaminoglycan used in the present invention can be produced by a conventionally known method. The sulfated glycosaminoglycan may be used in a purified form or in a proteoglycan form. In order to obtain sulfated glycosaminoglycan, for example, chondroitin sulfate in the form of proteoglycan, for example, sharks, marine animals such as whales, cattle, cartilage such as mammals such as pigs or birds as a raw material, neutral salt method, Extraction may be performed according to a known method such as an alkali method, an enzymatic method, or an autoclave method to remove fats and solids, and then dried. Furthermore, after removing fats and solids, the protein is further deproteinized using a proteolytic enzyme, and purified according to a known method by alcohol precipitation, so that chondroitin sulfate or a salt thereof can be obtained in a purified state.

The most preferred of the sulfated glycosaminoglycans used in the present invention is chondroitin sodium sulfate, which is extracted from fish and / or mammalian cartilage, and contains sodium chondroitin sulfate as a main component or only one. It is contained as glycosaminoglycan. Sharks are preferred as fish, and bovines are preferred as mammals.

Among them, sodium chondroitin sulfate having the following properties (1) and (2) is extremely preferred. (1) Nitrogen content when dried is determined to be 2.5 to 3.8
%. (2) Sulfur content of 5.5 ~
7.5%.

Further, those having the following properties (3) to (9) are particularly preferable. (3) When 100 mL of water was added to 1.0 g of this and dissolved, the solution was colorless to clear and slightly yellow. (4) The chloride content is 0.142% or less. (5) The sulfate content is 0.24% or less. (6) The heavy metal content is 20 ppm or less. (7) Arsenic content is 2 ppm or less. (8) Loss on drying is 10.0% or less (1 g, 105 ° C, 4 hours). (9) Residue on ignition is 23.0-31.0% (1 g, after drying).

The above (1) to (9) are performed according to the general test method described in “13th Revised Japanese Pharmacopoeia Commentary (Hirokawa Shoten)” and the method described in “Non-Japanese Pharmacopoeia Drug Standard 1997”. Can be measured.

The administration route of the agent for preventing and / or treating intestinal diseases of the present invention is not particularly limited, but is administered orally or enema because it is administered for the purpose of preventing and / or treating intestinal diseases. It is preferably administered orally, and particularly preferably administered orally.

The dosage form or administration form of the preventive and / or therapeutic agent for enteric diseases of the present invention includes tablets, capsules, granules, powders, liquids and the like, which are suitable for oral administration. Suppositories, liquid preparations and the like are also suitable for enema administration.

The above-mentioned dosage forms (formulations) of the agent for preventing and / or treating intestinal diseases of the present invention can be produced by a method commonly used in the art, for example, the above-mentioned sulfated glycosamino. Formulations containing glycans together with solid or liquid pharmaceutical carriers or diluents, ie, excipients, stabilizers, disintegrants and other additives can be formulated. These dosage forms include, for example, water, gelatin, lactose, starch, magnesium stearate, talc, animal and vegetable oils, benzyl alcohol, polyalkylene glycols, petroleum resins, coconut oil, lanolin, and the like, together with 0.1 g of sulfated glycosaminoglycan. It can be prepared so as to contain about 90 to 90% by weight.

The agent for preventing and / or treating intestinal diseases of the present invention may contain other pharmaceutically active substances in addition to the above-mentioned pharmaceutical carriers and the like. Such drugs include steroids, salazosulfapyridine, 5-aminosalicylic acid, and the like.

The dose of the agent for preventing and / or treating intestinal diseases of the present invention should be appropriately selected according to the type of the disease, the condition, age, weight, etc. of the patient, but generally, sulfuric acid contained as an active ingredient is used. Of sulfated glycosaminoglycan in an amount of about 20 to 500 mg, preferably 50 to 250, in terms of weight of 1 kg of body weight and one day as weight of the glycosaminoglycan.
It is preferable to administer about mg. When sulfated glycosaminoglycans are administered as proteoglycans,
What is necessary is just to make it the amount of sulfated glycosaminoglycan in a proteoglycan be in the said range.

The intestinal disease to which the preventive and / or therapeutic agent for intestinal disease of the present invention is applied is not particularly limited, but is preferably applied to inflammatory bowel disease, particularly to inflammatory bowel disease accompanied by bleeding. Is particularly preferred. Intestinal diseases that can be prevented and / or treated by the agent for preventing and / or treating intestinal diseases of the present invention include, but are not limited to, inflammatory bowel diseases such as ulcerative colitis and Crohn's disease, infectious enteritis, and ischemic disease. Enteritis, radioactive enteritis, intestinal behcet, simple (non-specific) intestinal ulcer, non-specific multiple small intestinal ulcer, acute ileitis, etc. Among these, application to ulcerative colitis, which is an inflammatory bowel disease accompanied by bleeding, is preferred.

The sulfated glycosaminoglycans, which are the active ingredients of the prophylactic and / or therapeutic agent for intestinal diseases of the present invention, are all biological substances and have been used in human bodies in various applications. It is considered that the preventive and / or therapeutic agent for intestinal diseases has sufficient safety even when administered to a living body.

[0027]

EXAMPLES Hereinafter, the present invention will be described in detail with reference to examples. Example Colitis was induced by dextran sodium sulfate in an animal model as described below, and the effect of chondroitin sulfate on it was examined and compared with the effect of 5-aminosalicylic acid, a known anti-inflammatory drug.

Materials and Methods Test substance 1) Inducing substance Dextran sulfate sodium 5000 (molecular weight 5000, Wako Pure Chemical Industries, hereinafter abbreviated as "DSS")

2) Test substance The chondroitin sodium sulfate used as the test substance was extracted by treating the shark cartilage with a proteolytic enzyme by a conventional method, removing fats and solids, and further removing the proteolytic enzyme. And then purified by alcohol precipitation.

The product thus obtained had the following properties. (1) Nitrogen content when dried is determined: 3.40% (2) Sulfur content when dried is determined: 6.82% (3) 1.0 g of this was dissolved in 100 mL of water (4) Chloride content: 0.142% or less (5) Sulfate content: 0.24% or less (6) Heavy metal content: 20 ppm or less (7) Arsenic content: 2 ppm or less (8) Loss on drying: 5.67% (1 g, 105 ° C, 4 hours) (9) Residue on ignition: 26.3% (1 g, after drying)

This was diluted to a predetermined concentration with distilled water for injection and used (hereinafter referred to as "CS").

As a control drug, 5-aminosalicylic acid (Wako Pure Chemical Industries, hereinafter abbreviated as "5-ASA") suspended at a predetermined concentration in 5% sodium carboxymethylcellulose (Wako Pure Chemical Industries) was used.

2. Animals 7-week-old male Wistar rats (Charles River Japan, S
PF).

3. Group composition

[0035]

[Table 1]

4. Methods Rats were individually housed in quadruple gauges and allowed to freely drink a 3% DSS aqueous solution daily. The test substance was orally administered once a day from the start of DSS administration to the day before the dissection. The control group received the same volume of distilled water instead of the test substance. The untreated group was allowed to freely drink purified water. Ten days after the start of DSS administration, the rats were anesthetized with ether, and blood was collected from the abdominal vena cava. After sacrifice, the lower colon and rectum (7 cm) were removed. The isolated intestine was fixed with 10% neutral buffered formalin, and then stained with Caracci hematoxylin, photographed, and the erosion area was measured using an image analyzer.

Blood tests such as blood cell count were performed using an automatic blood cell analyzer (Sysmex K-2000; manufactured by Sysmex).

The body weight and general symptoms (the state of feces such as bloody stool) were measured every day, and the amount of drinking water was measured and observed every other day. The bloody stool was scored and expressed as 0: normal, 1: bloody stool (blood adhered to the stool), and 2: bloody stomach (continuous bleeding from the anus).

5. Statistical processing Dunnett's multiple comparison test was used for comparison with the control group.

Results 1. Blood test values The results of blood white blood cell count, blood red blood cell count, and hematocrit value are shown in FIGS.

As for the white blood cell count in the blood, the control group showed about a three-fold increase as compared with the untreated group. The CS group had no significant effect compared to the control group, but showed a low value. The positive control 5-ASA also showed a significant inhibitory effect (FIG. 1).

With respect to the blood red blood cell count, a clear decrease in the number of red blood cells was observed in the control group, and a decrease of about 20% was observed in comparison with the untreated group. In the 100 mg / kg CS group and the 5-ASA group, this erythrocyte reduction was significantly suppressed (FIG. 2).

With respect to the hematocrit value, a clear decrease was observed in the control group, and a decrease of about 20% was observed in comparison with the untreated group. In CS 100 mg / kg group and 5-ASA group,
This decrease was significantly suppressed (FIG. 3).

2. Bloody stool The results of the bloody stool score are shown in FIG.

In the control group, observation was made 3 days after the start of DSS administration, and on the day of dissection, observation was made in all animals in the control group. CS markedly suppressed the development of bloody stool, and was significant in the 20 mg / kg and 500 mg / kg groups 4 days after administration and in the 100 mg / kg group 4 to 7 days after administration compared to the control group. Suppression was observed. In the 5-ASA group, a significant effect was observed 10 days after the start of administration (FIG. 4).

3. Erosion area The results obtained for the erosion area are shown in FIG.

The 20, 100 and 500 mg / kg groups of CS, and 5-
The erosion area of the ASA group was lower than that of the control group,
It was significantly suppressed in the CS 100 mg / kg group and the 5-ASA group (FIG. 5).

As is evident from the above results, the sulfated glycosaminoglycan containing chondroitin sulfate significantly suppresses DSS-induced stool expression, decreased blood red blood cell count, decreased hematocrit value, and erosion, and the present invention It has been shown that the preventive and / or therapeutic agent for intestinal disease has an effect on mucosal injury of the digestive tract including inflammatory bowel disease due to effects such as mucosal protective action or anti-inflammatory action.

[0049]

The present invention provides a preventive and / or therapeutic agent for intestinal disease useful for preventing and / or treating gastrointestinal mucosal injury including inflammatory bowel disease.

[0050]

[Brief description of the drawings]

[0051]

FIG. 1 is a graph showing the results of the effects of sulfated glycosaminoglycans on blood leukocyte counts obtained in Examples. * Indicates p <0.05.

[0052]

FIG. 2 is a graph showing the results of the effects of sulfated glycosaminoglycans on blood red blood cell counts obtained in Examples. * Indicates p <0.05, ** indicates p <0.01.

[0053]

FIG. 3 is a graph showing the results of the effects of sulfated glycosaminoglycans on hematocrit obtained in the examples. * Indicates p <0.05, ** indicates p <0.01.

[0054]

FIG. 4 is a graph showing the effect of sulfated glycosaminoglycan on bloody stool, obtained in the examples. *
Indicates p <0.05.

[0055]

FIG. 5 is a graph showing the effect of sulfated glycosaminoglycan on the erosion area obtained in the example. * Indicates p <0.05.

Claims (5)

[Claims] 1. A preventive and / or therapeutic agent for intestinal disease containing a sulfated glycosaminoglycan or a pharmacologically acceptable salt thereof. 2. The preventive and / or therapeutic agent for intestinal disease according to claim 1, wherein the sulfated glycosaminoglycan is derived from sharks and / or cattle. 3. The preventive and / or therapeutic agent for intestinal disease according to claim 1, wherein the intestinal disease is inflammatory bowel disease. 4. The preventive and / or therapeutic agent for intestinal disease according to claim 3, wherein the inflammatory bowel disease is inflammatory bowel disease accompanied by bleeding. 5. The preventive and / or therapeutic agent for intestinal disease according to claim 4, wherein the inflammatory bowel disease associated with bleeding is ulcerative colitis.