JPH08143468A - Antiulcer agent - Google Patents

Abstract

PURPOSE: To obtain an antiulcer agent free from side effect by using a peptide derived from a lactoferrin and having a specific amino acid sequence as an active component. CONSTITUTION: This antiulcer agent contains >=0.1mg, preferably >=1mg (based on 1g of the agent) of a peptide having the amino acid sequence of formula I, formula II, formula III, etc., a peptide derivative, a peptide salt or a mixture of two or more kinds of the above compounds as an active component. The peptide can be produced e.g. by separating from a decomposition product of lactoferrin. It can be formed to an oral drug by conventional method or included in a food to obtain a functional food having the action of an antiulcer agent. It is producible in a mass and stable as a drug because of its heat-resistance, water-solubility and stability in an aqueous solution. Since peptide has antimicrobial action, it does not necessitate the use of an antiseptic agent in preparation.

Description

Detailed Description of the Invention

[0001]

This invention relates to an antiulcer agent. More specifically, the present invention relates to a new anti-ulcer agent having no side effect, which contains peptides derived from lactoferrin having a specific amino acid sequence as an active ingredient.

[0002]

2. Description of the Related Art An ulcer (peptic ulcer) is a loss of tissue of a certain depth in the mucous membrane and skin due to necrosis in the esophagus, stomach, duodenum, etc., which is primarily caused by hydrochloric acid and pepsin in gastric juice. As antiulcer agents used for treating ulcers, antacids having a gastric acid neutralizing action, anticholinergics having a gastric acid secretion inhibiting action, H ₂ blockers, proton pump inhibitors and the like are known.

Examples of antacids include sodium hydrogen carbonate, magnesium carbonate, aluminum hydroxide, and mixtures thereof. However, since the duration of action is short, it is necessary to take the drug frequently, and formulations containing magnesium. Then, there are drawbacks such as strong warming effect (Medicinal Journal Vol. 27, 2272, 1991). Conventional anticholinergics and H ₂ blockers (eg cimetidine)
Has a high acute toxicity level and side effects due to drug binding to receptors other than the affected area (for example, with cimetidine, hematological disorders, liver disorders, renal disorders, endocrine system disorders, psychiatric / neurological disorders, gastrointestinal disorders) And hypersensitivity) and interference with other agents (Pharmaceutical Journal, Vol. 27, 227)
2 pages, 1991, and the Japan Pharmaceutical Collection, edited by the Japan Pharmaceutical Information Center, Pharmaceutical Industry Bulletin, 517 pages, 199.
3 years).

Similarly, the conventional proton pump inhibitors are also known to have defects such as carotenoid production (Pharmaceutical Journal, Vol. 27, p. 2272, 1991).
Year). On the other hand, antiulcer agents containing proteins and peptides as active ingredients include secretin, somatostatin, calcitonin, human epidermal growth factor (hEGF), urogastrone, and milk-derived growth factor (MGF) (JP-A-1- 121300), κ-casein glycomacropeptide (GMP), enzymatic degradation products thereof, and novel peptides derived from the κ-casein glycomacropeptide fraction (JP-A-4-210647 and JP-A-5-6529).
5, JP-A-5-262793, and JP-A-5-262793.
No. 271092) is known.

Among these proteins and peptides, peptide preparations such as secretin, somatostatin, calcitonin, human epidermal growth factor (hEGF) have the drawback that they can be administered only by injection which causes pain and difficulty in continuous administration. There is (Pharmaceutical Journal Volume 27, 22)
72 pages, 1991). Further, although urogastrone is a peptide preparation that can be orally administered, it has a drawback that the production amount is limited and the production cost is high because the purified raw material is pregnant horse urine. In addition, there are inconveniences associated with side effects such as dry mouth, nausea, stomach discomfort, diarrhea, constipation, etc.
Pharmaceutical Industry Bulletin, page 517, 1993).

Therefore, there has been a long-felt need for an anti-ulcer drug that does not have the above-mentioned drawbacks and has no side effects. Lactoferrin is
It is an iron-binding protein that is present in body fluids of mammals including humans such as milk and saliva, tears and mucous secretions, and is known to have antibacterial action against harmful microorganisms such as Escherichia coli, Candida and Clostridia. [Journal of Pediatrics,
94, p. 1, 1979]. It is also known to have an antibacterial action against staphylococci and enterococci at a concentration of 0.5 to 30 mg / ml [Journal of Dairy Scie
nce), 67, 606, 1984].

The inventors of the present invention pay attention to the antibacterial activity of lactoferrin, and focus on mammalian lactoferrin, apolactoferrin and / or metal-saturated lactoferrin (hereinafter, these may be referred to as lactoferrins).
It was found that a substance obtained by hydrolyzing lactic acid with an acid or an enzyme has no undesired side effects (for example, antigenicity) and has stronger heat resistance and antibacterial property than undegraded lactoferrins, and already applied for a patent ( JP-A-5-3
20068).

Further, the inventors of the present invention isolated a peptide having a strong antibacterial activity from a degradation product of lactoferrin, or synthesized a peptide having the same amino acid sequence as those peptides or a derivative of these peptides, Antibacterial peptide consisting of 20 amino acid residues (JP-A-5-92994), antibacterial peptide consisting of 11 amino acid residues (JP-A-5-78392), consisting of 6 amino acid residues Antibacterial peptide (JP-A-5-148297), antibacterial peptide consisting of 5 amino acid residues (JP-A-5-1498296), antibacterial peptide consisting of 3 to 6 amino acid residues (JP-A-5-29187) Nos. 5 to 148295) have already been applied for patents.

Furthermore, the inventors of the present invention have a brain-protecting action on peptides having the same amino acid sequence as the substance obtained by hydrolyzing lactoferrins with an acid or an enzyme or derivatives of these peptides (JP-A-6-172200). ) And nerve growth factor production promoting action (JP-A-5-32)
557), and already applied for a patent.

As another conventional technique, a method for separating and purifying lactoferrin from milk by utilizing the property that lactoferrin binds to heparin (JP-A-63-255299) has been disclosed. (JP-A-1-93534) and lactoferrin hydrolyzate have a digestive tract cell proliferation effect (JP-A-6-48955).

However, it is not known that these peptides derived from lactoferrins have a stronger antiulcer action as compared with lactoferrin and lactoferrin hydrolysates, and they are not described in the literature.

[0012]

As is clear from the above-mentioned prior art, an anti-ulcer agent that can be orally administered and has few side effects has been long sought, but still excellent substances are:
It was the current state of being unknown. The inventors of this invention have been earnestly researching more effective drugs,
We have found that peptides derived from lactoferrins have an anti-ulcer effect in vivo, and completed the present invention.

The present invention has been made in view of the above circumstances, and an object thereof is to provide an anti-ulcer agent which has few side effects and can be orally administered.

[0014]

Means for Solving the Problems The present invention is provided to solve the above-mentioned problems by providing peptides having an amino acid sequence set forth in any one of SEQ ID NO: 1 to SEQ ID NO: 31 and pharmaceutically acceptable peptides thereof. Derivatives, pharmaceutically acceptable salts of these peptides (hereinafter, these may be collectively referred to as peptides), or 2 of these
Provided is an anti-ulcer agent containing a mixture of one or more species as an active ingredient.

In the anti-ulcer agent of the present invention, it is also desirable that the active ingredient peptide is contained in an amount of at least 0.1 mg per 1 g of the active ingredient in the anti-ulcer agent. There is. In addition, at least 1 mg of the active ingredients peptides per 1 g
It is more desirable that the above content is included. Hereinafter, the configuration and preferred embodiments of the present invention will be described in detail.

When the peptides, which are the active ingredient of the anti-ulcer agent of the present invention, are produced from lactoferrins, the lactoferrins used as starting materials are commercially available lactoferrins and mammals (eg, human, bovine, sheep, goat,
Lactoferrin isolated from colostrum of horses, etc.), transitional milk, normal milk, end-stage milk, etc., or skim milk, whey, etc., which are processed products of these milk by a conventional method (for example, ion exchange chromatography) Apolactoferrin deironed with hydrochloric acid, citric acid, etc.
A metal-saturated or partially-saturated lactoferrin chelated with a metal such as copper, zinc, or manganese, which may be a commercially available product or a preparation prepared by a known method may be used.

The peptides used in the present invention are:
A peptide obtained by a separation means from a degradation product of lactoferrin, a peptide having the same amino acid sequence as this peptide, a peptide having a homologous amino acid sequence, a derivative of these peptides, a pharmaceutically acceptable salt of these peptides, or any of these And can be chemically synthesized by a known method. These peptides are
For example, JP-A-5-199294 and JP-A-5-19924.
78392, JP-A-5-148297, JP-A-5-1498296, and JP-A-5-148.
It can be obtained by the method described in each invention of Japanese Patent Publication No. 295.

The peptide obtained by the above method is
Peptides having the following amino acid sequences, their derivatives or salts can be exemplified as desirable embodiments. For example, peptides having the amino acid sequences of SEQ ID NOs: 1, 2 and 27,
Salts or derivatives thereof (JP-A-5-78392), peptides having the amino acid sequences of SEQ ID NOs: 3, 4, 5 and 6, salts or derivatives thereof (JP-A-5-78392).
No. 148297), SEQ ID NOS: 7, 8, 9 and 31
Having the amino acid sequence of SEQ ID NO: 5, a salt thereof or a derivative thereof (JP-A-5-1498296), a peptide having an amino acid sequence of any of SEQ ID NOS: 10 to 21, a salt thereof or a derivative thereof (JP-A-5-14829).
5), peptides having the amino acid sequences of SEQ ID NOs: 22 to 26, 28, 29 and 30; salts thereof or derivatives thereof (JP-A-5-92994).

Examples of the pharmaceutically acceptable salts of the above peptides include acid addition salts such as hydrochloride, phosphate, sulfate, citrate, lactate, tartrate, and the like. The derivative is a carboxyl group. Examples thereof include amidated or acylated amino group. The obtained peptides have extremely low toxicity as shown in Test Example 3 and can be appropriately used as an oral drug. According to known methods, tablets, capsules, troches, syrups, granules, powders, etc. It is also possible to process it. It is also possible to incorporate peptides as an active ingredient into foods and process them into antiulcer functional foods as one aspect of antiulcer agents. Of course, the anti-ulcer agent of the present invention also includes the improvement or prevention of gastric mucosal lesions such as so-called hemorrhagic gastritis, erosive gastritis and superficial gastritis as a prescription target.

The antiulcer agent of the present invention can be orally administered at a rate of at least 1 mg per 1 kg of body weight, although it varies depending on age, symptoms and the like. Next, the present invention will be described in more detail with reference to test examples. Test Example 1 This test was conducted to examine the antiulcer effect of peptides. 1) Test Animal Seven-week-old Donryu male rats (purchased from Japan SLC) having a body weight of 240 to 270 g were used by randomly dividing them into 5 groups (6 animals per group). 2) Test Agent As an anti-ulcer agent of the present invention containing peptides as active ingredients, the peptide of SEQ ID NO: 26 prepared by the same method as in Reference Example 1 was dissolved in distilled water to a concentration of 0.5 mg / ml. I used one. As a control drug, a substance prepared by dissolving stomatidine (a formulation containing cimetidine 40% (by weight) as an active ingredient, manufactured by Taisho Pharmaceutical Co., Ltd.) in distilled water at 1.25 mg / ml (0.5 mg / ml as cimetidine) was used. Distilled water was used as the blank test drug. 3) Test method A rat fasted for 48 hours was subjected to laparotomy under ether anesthesia.
Shay et al. Method for making pyloric ligation ulcer [Gastroenterology, Vol. 5, 43]
Pp. 61, 1945], the junction between the gastric pylorus and the duodenum was ligated and the abdomen was closed using a surgical adhesive and automatic suture forceps. Immediately after closure of the abdomen, the dose of the test drug was 1 mg / kg body weight or 10 mg / kg
Oral administration was performed so that the body weight was reached. For the blank test, distilled water was orally administered at a dose of 5 ml. After administration of each test drug, the animals were fasted for 13 hours and allowed to stand under water-free condition, then killed by exsanguination, and the stomach was excised and fixed by injecting 10 ml of 2% formalin solution. The formalin-fixed stomach was subjected to a large incision, and the degree of ulcer was observed. The degree of ulcer was expressed as the degree of mucosal damage by measuring the total area of the mucosal injury site per animal as described below and scoring in 6 stages. The cumulative frequency of mucosal damage was calculated as the sum of the mucosal damage levels (scores) of 6 animals in each test group, as a measure of the degree of ulcer in each test group.

Mucosal damage degree (score) Total area of mucosal damage site (mm ² ) 0 0 1 1 to 10 2 11 to 20 3 21 to 30 4 31 to 40 5 41 to perforation example 4) Test result 4) Test result Is as shown in Table 1. As is apparent from Table 1, the anti-ulcer drug administration group of the present invention containing the peptide of SEQ ID NO: 26 produced by the same method as in Reference Example 1 was compared with the control drug administration group (stomatidine) having a strong anti-ulcer effect. And showed the same remarkable mucosal damage inhibitory effect. From this test result, it was confirmed that the peptide produced by the same method as in Reference Example 1 had an anti-ulcer effect. In addition, no undesirable side effects such as allergic reaction occurring outside the affected area were observed by using the antiulcer agent of the present invention. The other peptides were also tested, but almost the same results were obtained.

[0022]

[Table 1]

Test Example 2 This test was carried out in order to compare the antiulcer effects of peptides, lactoferrin and lactoferrin degradation products. 1) Test animal 7-week-old Donryu male rats (purchased from Japan SLC) having a body weight of 240 to 270 g were randomly divided into 7 groups (6 animals per group) and used. 2) Test Agent As an anti-ulcer agent of the present invention containing peptides as active ingredients, the peptide of SEQ ID NO: 26 prepared by the same method as in Reference Example 1 was dissolved in distilled water to a concentration of 0.5 mg / ml. I used one. As the anti-ulcer agent containing lactoferrin as an active ingredient, commercially available bovine lactoferrin (manufactured by Sigma) was dissolved in distilled water to a concentration of 0.5 mg / ml. As an anti-ulcer agent containing a decomposed product of lactoferrin as an active ingredient, a bovine lactoferrin hydrolyzate prepared by the following method was used by dissolving it in distilled water at a concentration of 0.5 mg / ml. Distilled water was used as the blank test drug. (Preparation Method of Bovine Lactoferrin Hydrolyzate) 5 g of commercially available bovine lactoferrin (manufactured by Sigma) was added to purified water 90
Dissolve in ml, adjust the pH to 2.5 with 0.1N hydrochloric acid, and then commercially available pig pepsin (manufactured by Sigma) 100 mg
Was added and hydrolyzed at 37 ° C. for 6 hours. Then 0.1
Adjust the pH to 7.0 with regular sodium hydroxide and
Heat at 10 ° C for 10 minutes to inactivate the enzyme, cool to room temperature,
About 90 ml of bovine lactoferrin hydrolyzate was obtained. The bovine lactoferrin hydrolyzate was concentrated, freeze-dried, and powdered bovine lactoferrin hydrolyzate about 4.6 g.
I got 3) Test method The test was carried out by the same method as in Test Example 1. 4) Test results The results of this test are shown in Table 2. As is clear from Table 2, the peptide produced by the same method as in Reference Example 1 exhibited an antiulcer effect approximately 10 times that of bovine lactoferrin and a degradation product of bovine lactoferrin. From this test result, it was confirmed that the peptide produced by the same method as in Reference Example 1 had a stronger anti-ulcer effect as compared with bovine lactoferrin and a degradation product of bovine lactoferrin. Note that other lactoferrin, other lactoferrin degradation products, and other peptides were also tested, but almost the same results were obtained.

[0024]

[Table 2]

Test Example 3 This test was conducted to examine the acute toxicity of peptides. 1) Animals used Six-week-old CD (SD) strain rats (purchased from Japan SLC) were used, and males and females were randomly divided into 3 groups (5 animals per group) and subjected to the test. . 2) Test method The peptide of SEQ ID NO: 26 produced by the same method as in Reference Example 1 was added to water for injection (Otsuka Pharmaceutical Co., Ltd.) at 25 mg / m 2 respectively.
It was dissolved at a concentration of 1 and 50 mg / ml, and a single oral gavage was administered to each group of rats at a rate of 4 ml per 100 g of body weight using a needle with a metal ball (ratio of 1000 mg and 2000 mg per 1 kg of rat body weight). Acute toxicity was tested. As a blank test, water for injection (manufactured by Otsuka Pharmaceutical Co., Ltd.) was similarly administered to each one group of male and female rats and tested. 3) Test results The results of this test are shown in Table 3. As is clear from Table 3, no deaths were observed in the group to which this peptide was administered at the rates of 1000 mg / kg body weight and 2000 mg / kg body weight, as in the blank test (0 mg / kg). Therefore, the LD ₅₀ of this peptide is 2000
It was found to be more than mg / kg body weight and toxicity was extremely low. Similar tests were performed on other peptides, but almost the same results were obtained.

[0026]

[Table 3]

Test Example 4 This test was conducted to investigate the effective dose of peptides. 1) Test animal 7-week-old Donryu male rats (purchased from Japan SLC) having a body weight of 240 to 270 g were randomly divided into 4 groups (6 animals per group) and used. 2) Test drug As a test drug (an anti-ulcer agent of the present invention containing peptides as an active ingredient), the peptide of SEQ ID NO: 26 produced by the same method as in Reference Example 1 was added to distilled water at 0.5 mg /
What was melt | dissolved in the density | concentration of ml was used. Distilled water was used as the blank test drug. 3) Test method Test Example except that the peptide of SEQ ID NO: 26, which is an active ingredient in the test drug, was orally administered at a dose of 0.5 mg / kg body weight, 1 mg / kg body weight or 10 mg / kg body weight. Tested by the same method as 1. 4) Test results The results of this test are shown in Table 4. As is clear from Table 4, the peptide produced by the same method as in Reference Example 1 was orally at least 1 m / kg body weight.
The administration of g showed a remarkable antiulcer effect.
From this test result, it was confirmed that the effective oral dose of the peptide produced by the same method as in Reference Example 1 was at least 1 mg per 1 kg of body weight. Similar tests were performed on other peptides, but almost the same results were obtained.

[0028]

[Table 4]

Test Example 5 This test was carried out in order to examine the necessary amount of peptides to be incorporated in the antiulcer agent of the present invention. 1) Test Animal Seven-week-old Donryu male rats (purchased from Japan SLC) having a body weight of 240 to 270 g were used by randomly dividing them into 5 groups (6 animals per group). 2) Test drug As a test drug (antiulcer agent of the present invention containing peptides as an active ingredient), the peptide of SEQ ID NO: 26 produced by the same method as in Reference Example 1 was added to distilled water in an amount of 0.05 mg.
/ Ml, 0.1 mg / ml, 0.5 mg / ml or 1
What was melt | dissolved in each concentration (blending amount) of mg / ml was used. Distilled water was used as the blank test drug. 3) Test method For each concentration of the above-mentioned test agents, the dose of the peptide of SEQ ID NO: 26, which is the active ingredient in each test agent, is 1 mg / k.
The test was carried out by the same method as in Test Example 1 except that the dose was orally administered in an amount of g body weight. 4) Test results The results of this test are shown in Table 5. As is clear from Table 5, the dose was fixed to the lowest effective dose shown in Test Example 4 (ratio of 1 mg per 1 kg of body weight), and the compounding amount of the peptide produced by the same method as in Reference Example 1 was 0. 0.05 mg / ml, 0.1 mg / ml, 0.5 mg / ml
When it is set to ml or 1 mg / ml, the blending amount is 0.1 m
g / ml (If the specific gravity of distilled water is 1, 0.1 mg / ml
g) The above showed an anti-ulcer effect. Furthermore, the compounding amount is 1 m
g / ml (1 mg / g if the specific gravity of distilled water is 1)
The above showed a remarkable antiulcer effect. From this test result,
It was confirmed that the peptide, which is the active ingredient, produced by the same method as in Reference Example 1 should be contained in the anti-ulcer agent in an amount of at least 0.1 mg per 1 g.

Similar tests were conducted on other peptides, but almost the same results were obtained.

[0031]

[Table 5]

Reference Example 1 50 mg of commercially available bovine lactoferrin (manufactured by Sigma) was dissolved in 0.9 ml of purified water, pH was adjusted to 2.5 with 0.1N hydrochloric acid, and then commercially available porcine pepsin (manufactured by Sigma). (Manufactured by Mitsui Chemicals Co., Ltd.) and hydrolyzed at 37 ° C for 6 hours. Then, adjust the pH to 7.0 with 0.1 N sodium hydroxide, heat at 80 ° C. for 10 minutes to inactivate the enzyme, cool to room temperature, and centrifuge at 15,000 rpm for 30 minutes,
A clear supernatant was obtained. 100 μl of this supernatant was added to TSK gel O
It was subjected to high performance liquid chromatography using DS-120T (manufactured by Tosoh Corp.), and after injecting the sample at a flow rate of 0.8 ml / min, elution was performed with 20% acetonitrile containing 0.05% TFA (trifluoroacetic acid) for 10 minutes, and then. 0.0 for 30 minutes
Elution was carried out with a gradient of 20 to 60% acetonitrile containing 5% TFA, and the fractions eluted during 24 to 25 minutes were collected and dried under vacuum. This dried product was dissolved in purified water at a concentration of 2% (W / V), and the TSK gel ODS-120T was dissolved again.
(Manufactured by Tosoh Corporation) and subjected to high performance liquid chromatography at a flow rate of 0.8 ml / min for 10 minutes after injection of the sample.
Elute with 24% acetonitrile containing 05% TFA, then elute with a gradient of 24-32% acetonitrile containing 0.05% TFA for 30 minutes, 33.5-35.
Fractions eluting during 5 minutes were collected. The above operation was repeated 25 times, and vacuum drying was performed to obtain about 1.5 mg of the peptide.

The above peptide was hydrolyzed with 6N hydrochloric acid,
The amino acid composition was analyzed by an ordinary method using an amino acid analyzer. Identical sample is a gas phase sequencer (Applied
Edman degradation was performed 25 times using Biosystems) to determine the sequence of 25 amino acid residues. In addition, a disulfide bond analysis method using DTNB [5,5-dithio-bis (2-nitrobenzoic acid)] [Analytical B chemistry (Analytical B
iochemistry), vol. 67, p. 493, 1975]
Confirmed that a disulfide bond was present.

As a result, this peptide consisted of 25 amino acid residues, the 3rd and 20th cysteine residues were disulfide-bonded, and the 3rd cysteine residue resulted in N
-It was confirmed that the two amino acid residues on the terminal side have the amino acid sequence set forth in SEQ ID NO: 26 in which 5 amino acids were bonded to the C-terminal side from the 20th cysteine residue, respectively. It was Reference Example 2 Using a peptide automatic synthesizer (Pharmacia LKB Biotechnology Co., LKBBiolynx4170), a solid-phase peptide synthesis method by Shepard or the like [Journal of Chemical Society Perkin I (Jo
urnal of Chemical Society Perkin I), No. 538
Page, 1981], and the peptide was synthesized as follows.

An amino acid whose amine functional group is protected by a 9-fluorenylmethoxycarbonyl group [hereinafter sometimes referred to as Fmoc-amino acid or Fmoc-specific amino acid name (for example, Fmoc-asparagine)] has N, N-dicyclohexylcarbodiimide was added to produce the anhydride of the desired amino acid and this Fmoc-amino acid anhydride was used in the synthesis. An Fmoc-asparagine anhydride corresponding to the C-terminal asparagine residue was prepared to produce a peptide chain,
Through the carboxyl group, it is immobilized on Ultrosin A resin (Pharmacia LKB Biotechnology) using dimethylaminopyridine as a catalyst. The resin is then washed with dimethylformamide containing piperidine, C-
The protecting group of the amine function of the terminal amino acid is removed. Thereafter, Fmoc-arginine anhydride, which corresponds to the second position from the C-terminal of the amino acid sequence, was coupled to the deprotected amine functional group of arginine fixed to the resin via the C-terminal amino acid residue. In the same manner, glutamine, tryptophan, glutamine, and phenylalanine were sequentially fixed in the same manner. After the coupling of all amino acids is completed and the peptide chain of the desired amino acid sequence is formed, 94% T
The protecting groups other than acetamidomethyl were removed and the peptide was eliminated with a solvent consisting of FA, 5% phenol, and 1% ethanediol, the peptide was purified by high performance liquid chromatography, and the solution was concentrated and dried. The peptide powder was obtained.

The amino acid composition of the above-mentioned peptide was analyzed by an ordinary method using an amino acid analyzer, and SEQ ID NO: 10 was used.
It was confirmed to have the amino acid sequence described in 1.

[0037]

The present invention will be described in more detail and specifically with reference to the following examples, but the present invention is not limited to the following examples. Example 1 A tablet anti-ulcer drug having the following composition was produced by the following method. Lactose 18.8 (%) Corn starch 67.9 Magnesium stearate 1.4 Carboxymethylcellulose calcium 9.4 Peptide of SEQ ID NO: 26 2.5 Peptide of SEQ ID NO: 26 produced by the same method as Reference Example 1, lactose, To the mixture of corn starch and carboxymethyl cellulose calcium, sterilized purified water is appropriately added and kneaded uniformly, and dried at 50 ° C. for 3 hours,
Magnesium stearate was added to and mixed with the obtained dried product, and the mixture was tableted by a tableting machine according to a conventional method to give tablets. Example 2 An anti-ulcer syrup having the following composition was produced by a conventional method. Carboxymethyl cellulose calcium 0.20 (%) Sodium citrate 0.18 Citric acid 0.22 Fructose glucose liquid sugar 19.83 Purified water 78.57 Peptide of SEQ ID NO: 26 1.00

[0038]

INDUSTRIAL APPLICABILITY As described in detail above, the present invention relates to an antiulcer agent containing peptides as an active ingredient, and the effects achieved by the present invention are as follows. (1) Since it has few side effects, it can be administered for a long period of time. (2) It is heat-resistant, water-soluble, and stable in an aqueous solution, so that it is stable as a drug. (3) Since the peptide has an antibacterial action, it is not necessary to use a preservative for formulation. (4) It can be administered orally, and is simpler and more versatile than injections. (5) It is obtained from a relatively inexpensive raw material such as milk and can be mass-produced as compared with conventional peptide preparations.

[0039]

[Sequence Listing] SEQ ID NO: 1 Sequence length: 11 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment. In the sequence below, R01
Indicates any amino acid residue except Cys.

[0040] SEQ ID NO: 2 Sequence length: 11 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment. In the sequence below, R01
Indicates any amino acid residue except Cys.

[0041] SEQ ID NO: 3 Sequence length: 6 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment. In the sequence below, R01
Indicates any amino acid residue except Cys.

[0042] SEQ ID NO: 4 Sequence length: 6 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment. In the sequence below, R01
Indicates any amino acid residue except Cys.

[0043] SEQ ID NO: 5 Sequence length: 6 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment. In the sequence below, R01
Indicates any amino acid residue except Cys.

[0044] SEQ ID NO: 6 Sequence length: 6 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment. In the sequence below, R01
Indicates any amino acid residue except Cys.

[0045] SEQ ID NO: 7 Sequence length: 5 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment. In the sequence below, R01
Indicates any amino acid residue except Cys.

[0046] SEQ ID NO: 8 Sequence length: 5 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment. In the sequence below, R01
Indicates any amino acid residue except Cys.

[0047] SEQ ID NO: 9 Sequence length: 5 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment. In the sequence below, R01
Indicates any amino acid residue except Cys.

[0048] SEQ ID NO: 10 Sequence length: 6 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment.

[0049] SEQ ID NO: 11 Sequence length: 5 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment.

[0050] SEQ ID NO: 12 Sequence length: 4 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment.

[0051] SEQ ID NO: 13 Sequence length: 3 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment.

[0052] SEQ ID NO: 14 Sequence length: 5 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment.

[0053] SEQ ID NO: 15 Sequence length: 4 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment.

[0054] SEQ ID NO: 16 Sequence length: 4 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment.

[0055] SEQ ID NO: 17 Sequence length: 3 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment.

[0056] SEQ ID NO: 18 Sequence length: 6 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment.

[0057] SEQ ID NO: 19 Sequence length: 5 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment.

[0058] SEQ ID NO: 20 Sequence length: 4 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment.

[0059] SEQ ID NO: 21 Sequence length: 3 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment.

[0060] SEQ ID NO: 22 Sequence length: 20 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment. In the following sequence, Cys 2 and 19 are disulfide-bonded.

Sequence: Lys Cys Arg Arg Trp Gln Trp Arg Met Lys Lys Leu Gly Ala Pro 1 5 10 15 Ser Ile Thr Cys Val 20 SEQ ID NO: 23 Sequence Length: 20 Sequence Type: Amino Acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment. Cys * in the sequence below
Indicates cysteine in which a thiol group is chemically modified to prevent the formation of a disulfide bond.

Sequence: Lys Cys * Arg Arg Trp Gln Trp Arg Met Lys Lys Leu Gly Ala Pro 1 5 10 15 Ser Ile Thr Cys * Val 20 SEQ ID NO: 24 Sequence Length: 20 Sequence Type: Amino Acid Topology: Direct Chain Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment. In the following sequence, Cys 2 and 19 are disulfide-bonded.

Sequence: Lys Cys Phe Gln Trp Gln Arg Asn Met Arg Lys Val Arg Gly Pro 1 5 10 15 Pro Val Ser Cys Ile 20 SEQ ID NO: 25 Sequence Length: 20 Sequence Type: Amino Acid Topology: Linear Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment. Cys * in the sequence below
Indicates cysteine in which a thiol group is chemically modified to prevent the formation of a disulfide bond.

Sequence: Lys Cys * Phe Gln Trp Gln Arg Asn Met Arg Lys Val Arg Gly Pro 1 5 10 15

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification number Reference number within the agency FI Technical indication location C07K 5/08 8318-4H 7/06 ZNA 8318-4H (72) Inventor Hiroyuki Wakabayashi Zama City East, Kanagawa Prefecture 5-1-83 Morinaga Milk Industry Co., Ltd. Nutritional Science Laboratory (72) Inventor Minamitsuko Yamazaki 5-1-83 Higashihara, Zama City, Kanagawa Prefecture Morinaga Milk Industry Co., Ltd.

Claims (2)

[Claims] 1. A peptide having the amino acid sequence of any of SEQ ID NOs: 1 to 31, a pharmaceutically acceptable derivative of these peptides, a pharmaceutically acceptable salt of these peptides, or 2 thereof. An anti-ulcer agent containing a mixture of one or more species as an active ingredient. 2. The active ingredient is at least 0.
The anti-ulcer agent according to claim 1, which contains 1 mg.