JPH08231419A - Therapeutic agent for disease of upper-half digestive organ - Google Patents

Abstract

PURPOSE: To obtain a drug which is useful for prophylaxis or therapy of gastric ulcer, duodenal ulcer, chronic or acute gastritis, because the drug has an antiulcerative action. CONSTITUTION: This prophylactic or therapeutic drug for upper-half digestive diseases contains, as an active ingredient, hepatic parenchyma cell-growing factor, for example, 1) having an estimated molecular weight of about 76-92K according to the SDS-PAGE; 2) having an activity of proliferating hepatic parenchyma cells; 3) inactivating the activity by heat treatment at 80 deg.C for 10 minutes; 4) inactivating the activity by digestive treatment with trypsin or with chymotrypsin and 5) having strong affinity to heparin.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to the prevention and / or prevention of digestive disorders.
Or, it relates to a therapeutic agent. More specifically, the present invention relates to a medicament containing hepatic parenchymal cell growth factor as an active ingredient and useful for the prevention and / or treatment of upper gastrointestinal diseases such as gastric ulcer and gastritis.

The existence and properties of various growth factors have been reported so far, but it is known that certain growth factors have an antiulcer action. For example, EGF (epithelial cell growth factor) is orally administered (Gut, 29, 894, 1988) and subcutaneously (Scand. J.
Gastroenterol. Suppl., 162, 162, 1989),
It has been reported to promote healing of rat acetic acid ulcer, and it has been reported that EGF has a suppressive effect on the acute ulcer of rat by parenteral administration such as subcutaneous administration.
(Agents and Actions, 27, 294, 1989). Also, bFGF
Regarding (basic fibroblast growth factor), a healing promoting action on chronic duodenal ulcer generated in rats using cysteamine and an inhibitory effect on acute duodenal ulcer have been reported (Eur. J. Pharmacol., 183). , 2090, 1990). Recently, TGF-β (β subunit of a substance called tumor growth factor or transforming growth factor) promotes healing of acetic acid ulcer in rats, and gastric acid secretion suppressive action and cytoprotection are involved in the healing process. It has been reported that it is hardly involved (Japanese Patent Laid-Open No. 5-194259).

On the other hand, human hepatocyte growth factor (hereinafter sometimes abbreviated as “hHGF” in the present specification, and sometimes simply referred to as “HGF” when simply representing hepatocyte growth factor) is It was first isolated from plasma of patients with fulminant hepatitis as a human-derived protein factor capable of promoting the growth of primary cultured hepatocytes (Japanese Patent Laid-Open No. 63-22526). Then, the gene (cDNA) and amino acid sequence encoding the hHGF protein (JP
3-72883), a method for producing recombinant hHGF (JP-A-3-2828)
No. 5693) has been reported. Such recombinant human
HGF (hereinafter sometimes abbreviated as "rhHGF" in the present specification) is in vitro (J. Clin. Lnvest., 87, pp.1853-1.
857, 1991) and in vivo (Jpn. J. Pharmacol., 59, sup)
pl. 1, 137, 1992), it has the effect of promoting the proliferation and function of liver parenchymal cells.

Further, the target cells and target tissues of HGF have been widely searched, and HGF has various epithelial cells other than hepatocytes (tubular epithelium, lung epithelium, bile duct epithelium, gastric epithelium, etc.) It has been reported to change its proliferation and motility in response to lymphocyte cells (Mitsubishi Ka
sei R & D Review, 7, pp.16-24, 1993). Also, these HG
As a receptor molecule on F target cells, the proto-oncogene c-me
It has also been shown that the product is functional (Scien
ce, 251, pp.802-804, 1991). However, there is no report on the therapeutic effect or preventive effect of HGF on upper gastrointestinal diseases.

[0005]

DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention and Means for Solving the Problems An object of the present invention is to provide a preventive / therapeutic agent for digestive system diseases. More specifically, it is an object of the present invention to provide a medicament useful for preventing or treating ulcerative and / or inflammatory diseases of the upper gastrointestinal tract including the esophagus, stomach, and duodenum. As a result of diligent efforts to solve the above problems, the present inventor has found that HGF suppresses the development of upper gastrointestinal ulcers such as gastric ulcer and duodenal ulcer, and is effective in treating peptic ulcer. The present invention has been completed based on the above findings.

[0006] That is, according to the present invention, there is provided a prophylactic / therapeutic agent for upper gastrointestinal diseases, which comprises hepatocyte growth factor as an active ingredient. According to a preferred embodiment of the present invention, hepatocyte growth factor has the following physicochemical properties: 1) SDS-PAGE
Estimated molecular weight under non-reducing conditions is about 76,000-92,000
2) having the activity of proliferating hepatocytes; 3) 80
The above-mentioned prevention showing that the activity is inactivated by heating at 10 ° C for 10 minutes; 4) the activity is inactivated by digestion with trypsin and chymotrypsin; and 5) it has a strong affinity for heparin. .Therapeutic agents; and
There is provided the above preventive / therapeutic agent, wherein the upper gastrointestinal disease is an ulcerative and / or inflammatory disease of the esophagus, stomach and / or duodenum. Further, according to another aspect of the present invention, there is provided an anti-ulcer agent containing a hepatocyte growth factor or the above preferred hepatocyte growth factor as an active ingredient.

The drug of the present invention comprises a hepatocyte growth factor (HG
It is characterized by using F) as an active ingredient. As the hepatocyte growth factor, use is made of body fluids or tissues derived from mammals such as humans and rats known to contain HGF, or those isolated and purified from cells that spontaneously produce HGF. HGF can be
Recombinant HGF obtained by introducing the above cDNA into cells can also be used. Human-derived HGF (hHGF) is preferably used as the active ingredient of the medicament of the present invention.

The host for producing the recombinant HGF is not particularly limited, but for example, Escherichia coli, Bacillus subtilis, yeast, filamentous fungi, plant cells, insect cells, animal cells and the like may be used. More specifically, in order to produce a transformant having the ability to produce HGF, the placenta derived from the above mammals, liver tissue and blood of patients with liver damage, fibroblast cell lines such as MRC-5 cells and IMR-9 cells, Preferably, for host cells such as CHO cells, for example, JP
An expression vector containing a cDNA encoding HGF, preferably hHGF, may be introduced in accordance with the method described in Japanese Patent Publication No. 3-285693. It is a preferred embodiment of the present invention to use HGF isolated and collected by culturing such a transformant.

In addition to the above-mentioned natural or recombinant HGF itself, as an active ingredient of the medicament of the present invention, natural HG within the range of not impairing the precursor protein or the activity of growing hepatocytes.
Non-natural HGF in which a part of the amino acids of F is modified by substitution, deletion, insertion, modification or the like may be used. Examples of such non-natural HGF include JP2-288899, PCT International Publication WO90 / 10651, JP3-130091, JP3-255096, JP4-30000, Nature, 342, pp.440-443, 1
Those described in publications such as 989 can be used.

Particularly preferred HGF as an active ingredient of the medicament of the present invention has the following physicochemical properties: 1) The estimated molecular weight by SDS-PAGE (under non-reducing conditions) is about 7
6,000 to 92,000; 2) it has an activity to grow hepatocytes; 3) the above activity is inactivated by heat treatment at 80 ° C. for 10 minutes; 4) it is digested with trypsin and chymotrypsin. The activity is inactivated; 5) It has a strong affinity for heparin. As such HGF, those derived from human are more preferable, and hHGF specified by the amino acid sequence described in JP-A-3-72883 or JP-A-4-89499 is particularly preferable.

The pharmaceutical of the present invention comprises one or two of the above HGFs.
It is preferable to prepare one or more species alone or to prepare a pharmaceutical composition in the form of a formulation together with appropriate pharmaceutical additives and administer parenterally. The dosage form of such a pharmaceutical composition is not particularly limited as long as it is generally used for parenteral administration, and examples thereof include ampules for injection and freeze-dried powders for injection (filled in vials. ) Or the like can be used. Preparation into various dosage forms may be performed according to a method commonly used in the art, using well-known additive additives such as diluents and additives available in the art.

For example, a lyophilized powder for injection is prepared by dissolving an effective amount of the purified HGF in a diluent such as distilled water for injection, physiological saline and an aqueous solution of glucose, and if necessary, carboxymethylcellulose and sodium alginate. Excipients such as polyethylene glycol, sodium dextran sulphate, amino acids, stabilizers such as human serum albumin, preservatives such as benzyl alcohol, benzalkonium chloride, phenol, soothing such as glucose, calcium gluconate, procaine hydrochloride It can be produced by adding an agent, a pH adjusting agent such as hydrochloric acid, acetic acid, citric acid, sodium hydroxide and the like, and lyophilizing it according to a conventional method.

The ampoule for injection is prepared by dissolving an effective amount of the above HGF in a diluent such as distilled water for injection, physiological saline and Ringer's solution, and if necessary, a solubilizing agent such as sodium salicylate and mannitol, a quenching agent Sodium acid, buffer such as glycerin, isotonic agent such as glucose, added sugar, the above stabilizer, the above preservative, the soothing agent, the above pH
After adding an additive such as a regulator, it can be sterilized by ordinary heat sterilization, aseptic filtration, or the like to prepare. The sterilization method should be appropriately selected because it may be inactivated in the heat sterilization step depending on the type of active ingredient.

HGF, which is an active ingredient of the medicament of the present invention, has an antiulcer action in a hydrochloric acid / ethanol ulcer model. It is well known to those skilled in the art that a drug having such an action is effective for peptic ulcer such as gastric ulcer and inflammatory gastrointestinal diseases such as gastritis of mammals including human. Therefore, the medicament of the present invention includes, for example, ulcerative diseases of the upper gastrointestinal tract such as esophageal ulcer, gastric ulcer, and duodenal ulcer, diseases such as chronic and / or acute esophagitis, gastritis, or duodenitis, upper gastrointestinal perforation. It is effective for prevention and / or treatment of. The target of application of the drug of the present invention is not limited to the digestive system diseases of the above-mentioned specific diseases, and it is said that a disease in which two or more of these diseases are combined is also a target of application of the drug of the present invention. There is no end. From another viewpoint, the medicament of the present invention is useful as an antiulcer agent.

The pharmaceutical agent of the present invention may contain an active ingredient of another pharmaceutical agent having the same pharmacological action or other pharmacological action as the pharmaceutical agent of the present invention. In addition, active ingredients such as heparin, sulfated polysaccharides such as dextran sulfate or derivatives thereof (JP-A-5-301824), which are known to enhance the hepatocyte proliferation effect of the active ingredient of the medicament of the present invention, etc. You may mix | blend. Since these sulfated polysaccharides or their derivatives have the effect of enhancing the stability of the drug of the present invention, a drug containing them is a preferred embodiment of the present invention.

The medicament of the present invention is generally administered parenterally, more specifically by subcutaneous, intramuscular or intravenous injection, for the purpose of preventing and / or treating the above-mentioned diseases in mammals including humans. can do. In general, a given amount can be administered by injection in single or divided doses, or can be administered continuously by infusion. The dose should be appropriately increased or decreased according to the age, sex, symptoms, body weight, administration form, etc. of the patient, but in general, 1 μg / kg to 10 mg / kg per adult per day, more preferably 10 to 1000μ
It may be administered in the range of g / kg.

[0017]

The present invention will be described in more detail with reference to the following examples, but the scope of the present invention is not limited to these examples. Recombinant hHGF (rhHGF) produced according to the method described in JP-A-3-285693 is a solvent (6.25 mM
Sodium phosphate, 0.44 M NaCl, 0.5% BSA)
Donryu male rats (body weight around 200 g: fasted and dehydrated for 24 hours, 10 animals per group) were intraperitoneally administered at a dose of 1,000 μg / kg. 30 minutes after administration, hydrochloric acid / ethanol (hydrochloric acid 1:
Ethanol (50) was orally administered at 1 ml / rat, and 60 minutes later, the abdomen was opened and the stomach was fixed with 10 ml of 1% formalin, and the ulcer index (mm) was measured. The results are shown in Table 1.
It is clear that HGF significantly suppressed the development of disorders caused by the hydrochloric acid / ethanol ulcer model.

[Table 1]

EFFECTS OF THE INVENTION HGF, which is an active ingredient of the medicine of the present invention, has an anti-ulcer action, and has an upper gastrointestinal tract such as gastric ulcer, duodenal ulcer, esophageal ulcer, acute gastritis, chronic gastritis, duodenitis, gastrointestinal perforation, etc. It is useful for prevention and / or treatment of diseases.

Claims (3)

[Claims] 1. A prophylactic / therapeutic agent for upper gastrointestinal diseases, which comprises hepatocyte growth factor as an active ingredient. 2. The physicochemical properties of hepatocyte growth factor are as follows: 1) The molecular weight estimated by SDS-PAGE (under non-reducing conditions) is about 7.
6,000 to 92,000; 2) it has an activity to grow hepatocytes; 3) the above activity is inactivated by heat treatment at 80 ° C. for 10 minutes; 4) it is digested with trypsin and chymotrypsin. The activity is inactivated; 5) The preventive / therapeutic agent according to claim 1, which has a strong affinity for heparin. 3. The upper digestive system disease is an ulcerative and / or inflammatory disease of the esophagus, stomach, and / or duodenum.
Or the preventive / therapeutic agent according to 2.