JPH05271092A - Medicine and food additive for preventing or treating gastric ulcer - Google Patents

Abstract

PURPOSE:To provide a medicine and a food additive having not only an effect for preventing and treating gastric ulcer but also capability for massive administration for a long period in the state a little in the side-effects and used for a gastric ulcer-preventing or treating agent. CONSTITUTION:A gastric ulcer-preventing or treating agent contains a peptide fraction having a gastric acid secretion-inhibiting action and separated from human or mammalian milk as an active ingredient, and a food additive contains the fraction and has such the actions. Such peptide fraction includes a glucomacropeptide crude fraction obtained by allowing the milk to penetrate an ultrafiltration membrane having a fractional mol.wt. of 10000-50000 at a pH of <4 and subsequently concentrating the filtrate with an ultrafiltration membrane having a fractional mol.wt. of <=50000 at a pH of >=4, and further includes a peptide crude fraction obtained by allowing an anion exchange resin to absorb a glucomacropeptide crude fraction and subsequently making the absorbed substance elute. The peptide fractions are the ingredients of the milk and have gastric juice secretion-inhibiting actions.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a pharmaceutical and food additive for preventing or treating gastric ulcer, which comprises a peptide fraction having a gastric acid secretion inhibitory action isolated from human or mammalian milk as an active ingredient.

[0002]

2. Description of the Related Art Antiulcer agents include antacids that suppress the digestive action of gastric juice, anticholinergics, antigastrin agents, and histamine receptor antagonists that suppress gastric juice secretion itself. However, although antacids such as sodium bicarbonate have immediate effects, they have a short duration of action and induce alkalosis by long-term large-dose administration, and calcium carbonate preparations cause hypercalcemia that causes urolithiasis. However, magnesium has a drawback that diarrhea is likely to occur. In addition, anticholinergic drugs also have side effects such as dry mouth, constipation, and palpitations, so they cannot always be said to be satisfactory antiulcer drugs. further,
Peptide preparations, which are the mainstream of anti-gastrin agents, have to be purified from animal urine and organs, such as urogastrone and secretin, and therefore have high production costs and a limited production amount.

Recently, a histamine receptor antagonist has been developed, but it has a problem that ulcer is likely to recur when the administration is stopped. In a modern society where stress is particularly high, the lifetime morbidity rate of gastric ulcer is said to be 20%, and the probability of recurrence is very high (80% or more), and in the case of recurrence, administration of antiulcer drug must be restarted. That is, a patient who once suffers from gastric ulcer must continue to take an antiulcer drug, which is a pharmaceutical, on a regular basis, and even if a drug with few side effects is used, it cannot be said that there is no effect on the human body.

Under these circumstances, it is highly effective in suppressing the secretion of gastric juice, is not accompanied by the risk of side effects, and is an anti-ulcer drug that can be manufactured relatively inexpensively, or prevents the onset and recurrence of ulcers, which can be said to be a chronic disease. There is a strong demand for the provision of food materials with the functionality to do so.

[0005]

DISCLOSURE OF THE INVENTION The present invention solves the problems found in the conventional antiulcer agents as described above,
It was made in view of the characteristics of ulcer as a chronic disease. That is, the object is to provide a functional food additive for preventing the onset or recurrence of ulcer, which is obtained from a relatively inexpensive raw material, has a good gastric secretion inhibitory effect, and is effective for treating ulcer. To do.

Chernikov and Stan [21st International Daily Congress Volume 1 Book 2. 161
(1982)] conducted an experiment in which a pepsin degradation product of κ-casein was administered to the blood of dogs, and suggested that there are substances that suppress the secretion of gastric juice and substances that promote this secretion product. No specific experimental data is mentioned in the report. After that, there was no related report for a while, but in 1987 Guilloteau et al. [Reprod. Nutr. D
evelop.) 27 , 287-288 (1987)] reported that intravenous administration of casein macropeptides to calves did not alter gastric juice output. As described above, the present situation is that the gastric secretion inhibitory effect of the peptide obtained by enzymatically decomposing the milk protein has not yet been clearly concluded.

The inventors of the present invention have the following background.
Whey and whey protein concentrate produced during cheese manufacture (W
PC), deproteinized cheese whey, lactose mother liquor and other glycomacropeptides were collected and their pharmacological activities were examined. As a result, the raw fraction of glycomacropeptide obtained by ultrafiltration of these raw materials was gastric juice secreted. The inventors have found that it has an inhibitory action and completed the present invention.

[0008]

The present invention relates to a food additive containing a peptide fraction having a gastric secretion inhibitory action as an active ingredient, which can be added to a medicine or a food or drink for preventing or treating gastric ulcer.

The raw materials of the present invention include cheese whey or whey protein concentrate produced by ultrafiltration of cheese whey, or cheese whey obtained by precipitating and removing whey protein by a method such as heating, and lactose mother liquor. Can be used. In the present invention, glycos produced according to the methods described in JP-B-59-27358, JP-A-2-276542, JP-A-2-95686 and JP-A-63-284199. A crude macropeptide (GMP) fraction can be used as an active ingredient.

For example, as described in Japanese Patent Application Laid-Open No. 2-276542, raw milk, skim milk, etc. of humans, cattle, sheep, goats, etc. are used as raw materials, and cheese whey, WPC, removed from these raw materials are used. PH 4 for protein cheese whey, etc.
After adjusting to less than, ultrafiltration is performed using a membrane having a molecular weight cut off of 10,000 to 50,000 to obtain a filtrate. And this permeate is pH
A GMP crude fraction is obtained by adjusting to 4.0 or more and concentrating using a membrane having a molecular weight cutoff of 10,000 to 50,000. Alternatively, as described in Japanese Patent Application No. 3-19112, a pretreatment may be performed in which the raw material is brought into contact with an anion exchange resin and the adsorption fraction thereof is eluted. It is also possible to use the eluate obtained by adsorbing the GMP crude fraction on an anion exchange resin and eluting the adsorbed fraction with a buffer having an ionic strength of 0.25 or more. This eluate is a crude solution of peptide. At this time, the adsorption to the anion exchange resin is preferably carried out by dissolving the GMP crude fraction in a buffer solution containing 0.2 M of sodium chloride at pH 8.7 and adsorbing it on the anion exchange resin at pH 8.7. .. Then, the non-adsorbed portion is eluted using this buffer solution, and then the adsorbed fraction is converted to a buffer solution having an ionic strength of 0.25 or more
For example, the crude peptide fraction may be obtained by elution with a buffer solution containing 0.25 M or more, preferably 1 M sodium chloride and having a pH of 8.7. At this time, the anion exchange resin is Q-Sepharose, DEAE-Sepharose, QAE-Sephadex, QAE-Toyopearl, etc., and the buffer is Tris-HCl buffer, ethanolamine buffer, phosphate buffer, etc. Is used.

The crude peptide fraction thus obtained may be used as an antiulcer agent in a solution state, but it is usually dried by means of spray drying or freeze drying,
You may use it in a powder state. In addition, since these crude peptide fractions are stable to heat, it is more desirable to include a sterilization step before the drying step.

The crude peptide fraction obtained as described above can be used as an oral antiulcer agent such as sugar-coated tablets, tablets or capsules. In addition, various food and drink as food additives, for example, soft drinks, juice drinks,
By adding to fermented drinks, jellies, ice creams, etc., functional foods and drinks having an ulcer preventive or therapeutic effect can be obtained. Further, it can be added to sweets such as gum and candy.

The crude peptide fraction of the active ingredient in the present invention is a peptide derived from milk and has no adverse effect on the human body when orally ingested, and its intake amount is particularly limited. There is no. However, when actually ingested as an anti-ulcer drug or food for treating or preventing ulcer, 0.1 to 1000 mg / Kg body weight / day is appropriate,
Desirably, the dose is 1 to 100 mg / Kg body weight / day. That is,
No effect was observed at 0.1 mg / Kg body weight / day or less, and there was no side effect, but even if 1000 mg or more was administered, the effect was not markedly increased. The present invention is characterized in that the peptide can be administered in a large amount and continuously for a long period of time.

Of the active ingredients in the present invention, GM
The peptide crude fraction obtained by further purifying the P crude fraction with an anion exchange resin can also be used as a medicine such as an injection solution and other preparations. Although these crude fractions are said to have little antigenicity, even though they are milk-derived peptides, they are unlikely to cause allergic symptoms, etc., and the dose as an injection solution is 0.01 to 100 mg / Kg body weight. / Day, preferably about 0.1 to 10 mg / kg body weight / day. That is, 0.01 mg / Kg body weight / day or less has no effect, and 100 mg / Kg body weight / day or more has no side effects and no trouble, but the effect is not particularly increased.

In both oral administration and administration by injection, it may be administered once a day or in several divided doses.

[0016]

The action and effect of the antiulcer agent or the food additive having such action according to the present invention is summarized as follows. (1) Since it is a milk-derived peptide that is usually taken as food, there is no risk of side effects due to administration. (2) Since milk, which is a food, is used as a raw material, the manufacturing cost is very low compared to conventional antiulcer agents. (3) Since it can be prepared in a larger amount than conventional anti-ulcer agents,
Not only as a medicine, but also as a food additive, it can be widely used by adding it to various foods and drinks.

The present invention will be specifically described below based on Examples and Reference Examples.

[0018]

Reference Example 1 This example shows a method for preparing a peptide fraction having a gastric acid secretion inhibitory action in the present invention. Whey protein concentrate (Taiyo Kagaku, trade name Sanlacto N-
2) Dissolve 1 kg in 50 liters of water at 50 ° C and add pH 3.
After adjusting to 5, an ultrafiltration membrane with a molecular weight cut off of 20,000 (DD
S, GR61pp) ultrafiltered (pressure 0.4MPa)
, Average permeation flow rate 52.4 l / m ² · h). Permeate volume is 4
When it reached 0 liters, 40 liters of water at 50 ° C. was added to the concentrated solution and continuously subjected to ultrafiltration to obtain 160 liters of permeate. Add 25% sodium hydroxide solution to this permeate
The pH was adjusted to 7.0, and ultrafiltration was performed again using the same ultrafiltration membrane until the concentrated liquid reached 5 liters for desalting and concentration. Then 50 ℃
Water was added to the solution to maintain the concentration of the concentrated solution at 10 liters, and diafiltration was performed under the same conditions as before, for further desalting. When the amount of the permeated liquid reached 80 liters, water was not added to the concentrated liquid, and the concentrated liquid was concentrated to 2 liters and freeze-dried to obtain 54 g of a crude GMP fraction. This fraction was pure when analyzed by urea-SDS electrophoresis.
It was 82%.

[0019]

Reference Example 2 1.0 g of crude GMP fraction obtained in Reference Example 1
Is dissolved in 20 ml of 20 mM ethanolamine buffer of pH 8.7 containing 0.2 M sodium chloride, and this is passed through 200 ml of Q-Sepharose Fast Flow anion exchange resin.
Wash the resin with 600 ml to elute the non-adsorbed fraction. Next, pass 300 ml of 20 mM ethanolamine buffer of pH 8.7 containing 0.3 M sodium chloride and collect the eluted fraction. The eluted fraction was freeze-dried to obtain 50 mg of the crude peptide fraction of the present invention.
To get

[0020]

Example 1 This example shows the effect of suppressing gastric secretion in the intravenous administration test of the anti-ulcer substance according to the present invention. [Test Method] Wistar rats (male, 7 months old, 8 animals in each group) that had been fasted for 16 hours and restricted water supply for 2 hours,
GMP crude fraction according to Reference Example 1 dissolved in physiological saline
0.01, 0.1, 1, 10, 100 mg / Kg body weight, β-lactoglobulin 1 mg / Kg body weight or physiological saline alone was intravenously injected, and the gastric pyloric region was immediately ligated. Four hours later, the pyloric region of the stomach was also ligated to remove the stomach, and the gastric juice accumulated in the stomach was collected and the volume was measured, and the acidity was measured by titration. As shown in Table 1, gastric acid secretion was significantly reduced in the group to which the crude GMP fraction was administered at 0.1 mg / Kg body weight or more.

[0021]

[Table 1]

[0022]

Example 2 This example shows the effect of suppressing gastric secretion in the oral administration test of the antiulcer substance of the present invention. [Test method] For Wistar rats (male, 7 months old, 8 animals in each group) that had been fasted for 16 hours and had a limited water supply for 2 hours.
GMP crude fraction from Reference Example 1 dissolved in 2 ml of water
0.1, 1, 10, 100, 1000 mg / Kg body weight or water alone was orally administered, and 1 hour later, the gastric pyloric region was ligated. Four hours later, the gastric cardia was also ligated to remove the stomach, and the gastric juice accumulated in the stomach was collected and the volume was measured, and the acidity was measured by titration. As shown in Table 2, gastric acid secretion was significantly reduced in the group to which the crude GMP fraction was administered at 1 mg / Kg body weight or more.

[0023]

[Table 2]

[0024]

Example 3 This example shows the therapeutic effect on gastric ulcer in the intravenous administration test of the anti-ulcer substance according to the present invention. [Test Method] Wistar rats (male, fasted for 16 hours)
(8 weeks old, 6 animals in each group), 0.5 ml of 70% ethanol was orally administered, and 1 mg / kg body weight of crude peptide according to Reference Example 2 immediately dissolved in physiological saline and 1 mg of β-lactoglobulin
/ Kg body weight or physiological saline was injected intravenously. Five hours later, the stomach was removed, a large incision was made, the mucosal surface was washed with physiological saline, and the ulcer degree was observed. The ulcer degree was divided into 6 stages according to the degree of mucosal hemorrhage lesions. As shown in Table 3, clear reduction of ulcer was observed in the crude peptide fraction administration group.

[0025]

[Table 3] Ulcer degree: 0 = no ulcer is found in the gastric mucosa. 1 = Redness only. 2 = 1 bleeding erosion. 3 = 2-5 bleeding sores. 4 = 6-9 bleeding sores. 5 = 10 or more bleeding sores. (Average value ± standard deviation (n = 6)

[0026]

Example 4 This example shows an example in which the antiulcer substance of the present invention is used in a food for ulcer prevention. (1) Production of milk beverage for ulcer prevention Using the GMP crude fraction of Reference Example 1 as a food additive, a milk beverage for ulcer prevention was prepared according to the following formulation.

[0027]

[Table 4]

Based on the above composition, skim milk 23.1 g, glucose 15 g, vitamins 0.06 g, yogurt flavor 0.54
g, and 3 g of GMP crude fraction according to Reference Example 1 at 60 ° C.
Safflower oil 6 in a solution dissolved in 100 ml of warm water
g and sugar ester (trade name DKF160) 0.3 g at 60 ° C
The mixture obtained in (1) was gradually added dropwise with stirring using a TK homomixer to emulsify. This was heat-sterilized at 90 ° C for 5 minutes and then cooled to 10 ° C to obtain a product. In addition, from the above recipe, G
A milk drink except for MP was produced in the same manner. Next, these milk drinks were given to rats to examine the antiulcer effect.

[Test Method] A milk drink to which a food additive consisting of the GMP crude fraction of Reference Example 1 was added at the same time as the start of fasting,
Alternatively, to Wistar rats (male, 8 weeks old, 6 animals in each group), which had been allowed to freely drink a milk beverage produced by removing only the GMP crude fraction, 16 hours later, 70 ml of alcohol was administered in an amount of 0.5 ml each. Further, after 5 hours, the stomach was removed and a large incision was made. The mucosal surface was washed with physiological saline, and then the ulcer degree was observed. As shown in Table 2, clear reduction of ulcers was observed in the group administered with the milk drink containing the food additive consisting of the GMP crude fraction.

(2) Production of gastric ulcer-preventing jelly The GMP crude fraction of Reference Example 1 was produced by the following formulation to produce a ulcer-preventing jelly.

[0031]

[Table 5]

With the above formulation, sugar, gelatin, and the GMP crude fraction of Reference Example 1 were used as food additives, and 50 ml of water was added to them.
Was added and dissolved by heating at 80 ° C., prune extract was added thereto, and the mixture was stirred, transferred to a container and cooled. Also, G
A jelly containing no MP crude fraction was similarly prepared.

Next, these jellies were given to rats to
The preventive effect on gastric ulcer was examined. [Test Method] Wistar rats (male, fasted for 16 hours)
8 weeks old, 6 mice in each group), 2 g of jelly containing the above GMP crude fraction or jelly not containing the GMP crude fraction
Each was administered. One hour later, 0.5 ml of 70% alcohol was administered to each, and after 5 hours, the stomach was excised, a large incision was made, the mucosal surface was washed with physiological saline, and the ulcer degree was observed. Table 3
As shown in, the group to which the jelly containing the GMP crude fraction was administered showed a clear reduction in ulcer.

Claims (6)

[Claims] 1. A preventive or therapeutic agent for gastric ulcer comprising a peptide fraction having a gastric acid secretion inhibitory action isolated from human or mammalian milk as an active ingredient. 2. The peptide fraction has a molecular weight cut off at a pH of less than 4.
The agent according to claim 1, which is a crude fraction of glycomacropeptide obtained by permeation through an ultrafiltration membrane of 10,000 to 50,000 and desalting and concentrating using an ultrafiltration membrane having a molecular weight cutoff of 50,000 or less at pH 4 or more. 3. The peptide fraction has a molecular weight cut off at a pH of less than 4.
It permeates through 10,000 to 50,000 ultrafiltration membranes, is desalted and concentrated using an ultrafiltration membrane with a molecular weight cutoff of 50,000 or less at pH 4 or more, and is adsorbed on an anion exchange resin, and the adsorbed fraction has an ionic strength of 0.25 or more. The agent according to claim 1, which is a crude peptide fraction obtained by elution with the buffer solution of 1. 4. A food additive having a gastric ulcer-preventing or therapeutic action, which comprises a peptide fraction having a gastric acid secretion-suppressing action isolated from human or mammalian milk as a gastric acid secretion-suppressing component. 5. The peptide fraction has a molecular weight cut off at a pH of less than 4.
An additive according to claim 4, which is a crude fraction of glycomacropeptide obtained by permeation through an ultrafiltration membrane of 10,000 to 50,000 and desalting and concentration using an ultrafiltration membrane having a molecular weight cutoff of 50,000 or less at pH 4 or more. .. 6. The peptide fraction has a molecular weight cutoff at a pH of less than 4.
It permeates through 10,000 to 50,000 ultrafiltration membranes, is desalted and concentrated using an ultrafiltration membrane with a molecular weight cutoff of 50,000 or less at pH 4 or more, and is adsorbed on an anion exchange resin, and the adsorbed fraction has an ionic strength of 0.25 or more. 5. The additive according to claim 4, which is a crude peptide fraction obtained by elution with the buffer solution of 1.