JPH0812588A - Composition - Google Patents

Abstract

PURPOSE:To obtain a composition containing a culture filtrate of a cultured mycelium of Cordyceps militaris or an extract of the mycelium with a solvent, useful for producing medicines, foods and beverages, having excellent cardiac action and bronchodilatory action and high safety. CONSTITUTION:A mycelium of a fungus [e.g. Cordyceps militaris MF-20,005 (FERM P-14,398)] of the genus Cordyceps militaris is inoculated into a medium and subjected to shaking culture at 25 deg.C at 180rpm for 5 days. The culture product is centrifuged and separated into the mycelium and a filtrate. The culture filtrate and/or the mycelium is extracted with hot water at 85-140 deg.C to give a solvent extract wit a solvent, which is prepared into a medicine type and/or a food and beverage type to give the objective composition useful as a medicine, a health drink, etc., having excellent cardiac action and bronchodilatory action and high safety.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a composition having excellent pharmacological and health effects, and more particularly to a pharmaceutical type composition and / or a food / beverage type composition derived from Cordyceps militaris.

[0002]

BACKGROUND OF THE INVENTION Cordyceps is an ascomycete fungus.
cota), Pyrenomycetes,
Clavicipitales, Clavicipitaceae or Hypocre
acea), which belongs to the genus Cordyceps and has a complete era and an incomplete era.

The caterpillar of Cordyceps sinensis has been prized since ancient times for its fruiting body as a medicinal agent for longevity and as a nutritional tonic. In this way, the Cordyceps sinensis, which has been conventionally heavily used as a herbal medicine, is usually obtained by pulverizing the fruiting bodies and taking them.

On the other hand, the present invention is one in which a specific Cordyceps sinensis is cultivated and the obtained cultured mycelium is extracted with a solvent. Such a solvent extract has excellent cardiotonic action and antitussive action. This has never been known.

[0005]

DISCLOSURE OF THE INVENTION The present invention is intended for the purpose of newly developing a drug excellent in cardiotonic action and antitussive action or foods and drinks having these actions, and also for long-term administration from the viewpoint of safety. It was made for the purpose of developing a substance derived from a natural product so as to sufficiently endure it.

[0006]

[Means for Solving the Problems] As a result of examining from various aspects to solve the above problems, attention is focused on Chinese herbs, Cordyceps militaris, especially Cordyceps militaryalis, and after further research, its mycelium, and It was discovered that the solvent extract had excellent cardiotonic action and antitussive action, and as a result of further research based on this new finding, the present invention was finally completed. Hereinafter, the present invention will be described in detail.

In order to carry out the present invention, it is necessary to cultivate the mycelium of Cordyceps militaris belonging to the Cordyceps sinensis and to obtain the cultured mycelium. In the present invention, a microorganism belonging to Cordyceps militaryis (hereinafter sometimes referred to as the present bacterium) can be used, and for example, Cordyceps militaryis MF-20005 (Cordyceps mil) can be used.
Itaris MF-20005) is useful, and this strain was transferred to the Institute of Biotechnology, Institute of Industrial Science and Technology, FERM P
Deposited as -14398.

[0008] In the present invention, the mycelium of Cordyceps militaris is cultured.

Cultivation is carried out using a medium containing malt extract, yeast extract, peptone, broccoli broth, glucose, vitamins, amino acids, nucleic acids, proteins and, if necessary, host components such as insects, which are liquid or liquid. Sufficiently grow by solid culture.

In the case of large-scale culturing, it is preferable to carry out liquid culturing and aeration stirring culturing, pH 4 to 7, culturing temperature 1
5 to 32 ° C, preferably 20 to 30 ° C, 50 to 500r
It is preferable to carry out the culture for 3 to 10 days while slowly stirring at pm, preferably about 100 to 300 rpm.

After completion of the culture, the culture solution is filtered to separate into mycelium and culture filtrate. The mycelium is extracted with a solvent. As the solvent, water and an organic solvent are used. As the organic solvent, alcohols such as methanol, ethanol, propanol, butanol; methyl ether, ethyl ether,
Ethers such as propyl ether, methyl ethyl ether, and methyl propyl ether; ketones such as acetone, ethyl methyl ketone, methyl propyl ketone, and butyl methyl ketone are appropriately used, and a mixed solvent thereof and a mixed solvent with water can also be used. . Although the amount of the solvent is not particularly limited, it is preferably 1 ml to 1 L per 1 g of dry cells.

In the case of water extraction, the cultured mycelium is added to 85-140.
Extraction is performed under high temperature conditions of 90 ° C., preferably 90 to 125 ° C. In this case, water is added to the cultured mycelium, the mycelium is crushed if necessary, and then extraction is carried out under high temperature conditions with stirring if desired. Usually, hot water is used for extraction, but when extraction is performed at a higher temperature, autoclave extraction may be performed.

The solvent extract thus obtained is used as the active ingredient of the composition of the present invention after the extraction residue is removed, and then the liquid part is used as it is or the treated product is used. The processed product broadly includes a concentrate in the liquid part, a paste, a dried product and / or a diluent in the liquid part. If necessary, it is also possible to use the solvent extract as it is or to use the treated product without removing the extraction residue.

The culture filtrate or the solvent extract (including the treated product) of the mycelium has a cardiotonic action, an antitussive action and a bronchoconstriction inhibiting action, as will be apparent from the examples described below. The composition according to can be used as food and drink utilizing these actions, food and drink for specified health use, health drink, health food, nutritional food and other various types of food and drink, and also used as a medicine such as cardiotonic agent or antitussive. be able to.

When used as a food and drink type composition, the culture filtrate and / or the solvent extract (the processed product), which is the present active ingredient, can be used as it is or in combination with other foods or food ingredients. Or, it can be used according to an ordinary method. The composition according to the present invention using the present active ingredient may be in a solid form (powder, granules, etc.), paste form, liquid form or suspension form, but for the production of sweeteners, acidulants, vitamins and other drinks. It is preferable to formulate a health drink using various commonly used ingredients.

When used as a pharmaceutical type composition, the active ingredient is administered in various forms. Examples of the dosage form include tablets, capsules, granules, powders,
Oral administration such as syrup may be mentioned. These various preparations are commonly used in the pharmaceutical preparation technical field such as excipients, binders, disintegrants, lubricants, flavoring agents, solubilizing agents, suspending agents, coating agents, and the like, in accordance with the usual methods for the main drug. It can be formulated using known adjuvants. Although the amount used varies depending on symptoms, age, body weight, administration method, dosage form, etc., usually, about 0.1 mg to 1,000 mg can be administered once to an adult.

Since the active ingredient according to the present invention has a natural origin and has been used as a herbal medicine for many years, it has no or very low toxicity.
Shows excellent safety, 500 per day for rats
Oral oral administration (mg) was not observed, but no acute toxicity was observed.
Therefore, when used as a food-drink type composition, there is no particular limitation on the amount of the active ingredient used for prevention, health care, or when used as a food or drink,
Even when it is used as a medicine, it may be appropriately used within the above range depending on the patient. Further, since the present active ingredient does not exhibit remarkable acute toxicity even if it is taken in a large amount, it may be used in a larger amount than the above range if necessary.

The production examples and examples of the present invention will be described in detail below.

[0019]

[Production Example 1] M20Y2 medium (composition in 100 ml: malt extract (Oxoid) 2 g, yeast extract (Difc)
o) 0.2 g; pH 5.5) was dispensed into a 500 ml Erlenmeyer flask in 100 ml aliquots and placed in an autoclave for 121
Sterilized at 15 ° C for 15 minutes.

These sterilized media were added to Cordyceps
militaris MF-20005 (FERM
(P-14398) M20Y2 agar slant culture mycelium was inoculated with 1 platinum loop and cultivated with orbital shaking at 25 ° C. and 180 rpm for 5 days to obtain a culture. The culture was centrifuged and the separated mycelium and filtrate were freeze-dried.

The dried product of the filtrate was used as sample (1). For the mycelium, the dried product was divided into 2 minutes, and one of them was added and stirred with 20 ml of methanol and left standing overnight under refrigeration, and this filtrate was used as a sample (2). The remaining one was suspended in 40 ml of purified water, heated with a cooling tube at 90 ° C. for 2 hours for extraction, filtered, and the filtrate was freeze-dried to obtain a sample (3). The mycelium remaining after filtration is
Further, it was suspended in 40 ml of purified water, heated and extracted at 121 ° C. for 15 minutes, and the filtrate was freeze-dried to obtain a sample (4). (Fig. 1)

[0022]

[Example 1] SD of the culture filtrate of Cordyceps mycelium or the solvent extract of the mycelium produced in Production Example 1
Using the heart and bronchus isolated from the rat, the method of Kuroo et al. (Kurokawa M. and Tsunooo) was used.
A. , J. Physiol. , 407 , 135-15
3, 1988; Tsunoo A .; et al. ,
J. Physiol. , 433 , 163-181, 19
According to 91), its action was tested and its effectiveness was confirmed.

(1) Myocardial Specimens The relative values of the contractile force of the myocardium and the contraction interval time with respect to the right atrium of the male SD rat were determined by applying the culture filtrate and the solvent extract. The right atrium was extracted from a male SD rat (380 to 450 g) while maintaining the pacemaker function. The right atrium sample was fixed in a horizontal perfusion tank with a volume of 0.8 ml, and the contraction force was recorded isometrically. Perfusate is 95% oxygen, 5%
Krebs solution equilibrated with carbon dioxide, or a salt solution (sodium chloride 140 mM, aerated with the following composition)
Potassium chloride 5 mM, calcium chloride 2.6 mM, magnesium chloride 1.3 mM, glucose 10 mM, HEP
ES5mM) was used. The perfusate was kept at 36-37 ° C and the flow rate was 3-4 ml / min.

As a result, the culture filtrate and the solvent extract (10
0 μg / ml) increased the tension of contraction by the autonomic capacity of the right atrial muscle by 10 to 15% of the control. It did not have a prominent effect on the time interval between contractions by the automatic function.

(2) Bronchial specimen By applying the culture filtrate and the solvent extract, the effectiveness of the extract on contraction of male SD rat bronchus by electrical stimulation and the effectiveness of the extract on contraction by potassium were measured. did. A bronchus was extracted from a male SD rat to prepare a spiral specimen. The sample was fixed in the perfusion layer under the same conditions as above. A 50 mM potassium chloride solution for stimulation was similarly prepared.
The electrical stimulation was performed at intervals of 30 seconds through a platinum wire in the perfusion tank.

As a result, all solvent extracts other than the culture filtrate suppressed the transient contraction of the bronchi due to electrical stimulation (FIG. 2). The culture filtrate and the solvent extract showed a relaxing effect on the persistent contraction by 50 mM potassium (Table 1 below). In the relaxation rate in this case, the contraction value of 50 mM potassium is 100%, and the relaxation value is shown in%.

[0027]

[Table 1]

From the results of (1) and (2), the solvent extract of the present invention has a mild myocardial contractile force-enhancing effect and also suppresses transient and persistent contraction of the bronchi including the culture filtrate. Therefore, it is considered that the airway resistance is lowered and the efficiency of lung ventilation is increased. That is, it was confirmed that the composition according to the present invention has utility as a cardiotonic agent and an antitussive agent.

[0029]

Example 2 Methanol extract powder 10 obtained in Production Example 1
0 g, sugar 150 g, honey 15 g, ascorbic acid 1
g, 0.5 g of citric acid, and 1 kg of water by adding an appropriate amount of perfume to sterilize the mixture at 95 ° C. for 20 minutes, and aseptically filling 100 ml of each bottle into a food-drink type health drink.

[0030]

Example 3 2 of the methanol extract powder obtained in Production Example 1
200 g of 0% aqueous solution, 5 g of tocopherol acetate, 10 g of thiamine nitrate, 20 g of nicotinic acid amide, 50 g of anhydrous feine, 50 g of benzoic acid salt and a suitable amount of perfume and deionized water to make 30 L, and after sterilizing, aseptically fill each 30 ml bottle. And manufactured a health drink as a medicine.

[0031]

Example 4 (1) Material produced in Production Example 1 (2) 50 g (2) Lactose 90 g (3) Corn starch 29 g (4) Magnesium stearate 1 g (1), (2) and (3) (however, 17 g ),
Granulated with the paste prepared from (3) (however, 7 g). To the obtained granules, (3) (however, 5 g) and (4) were added and mixed well, and the mixture was compressed by a compression tableting machine to obtain 1000 tablets each containing 50 mg of the active ingredient (1). Manufactured.

[0032]

[Example 5] The same treatment as in Example 4 was carried out except that the substance (2) produced in Production Example 1 was replaced with the same (4),
Tablets were produced.

[0033]

Industrial Applicability According to the present invention, a composition having excellent stability derived from a natural product and having a gentle cardiotonic action and bronchodilator action or antitussive action is stable by adopting a culturing method even though it is derived from a natural product. And industrially supplied.

[Brief description of drawings]

1 shows a process for producing a culture filtrate and a solvent extract according to the present invention.

FIG. 2 shows the effect on transient contraction of the bronchus by electrical stimulation by applying the culture filtrate and the solvent extract according to the present invention. The vertical axis represents the ratio of the bronchoconstriction value to the control.

Claims (9)

[Claims] 1. Cordyceps militaryis (Cordy)
composition comprising a culture filtrate of a culture mycelium of Ceps militaris) and / or a solvent extract of the mycelium. 2. Corgiceptus militaryis MF-20
005 (Cordyceps militaris M
F-20005) is used.
The composition according to. 3. The composition according to claim 1 or 2, wherein the solvent is water or an organic solvent selected from alcohols, ethers and ketones. 4. The water extract gives cultured mycelium in an amount of 85 to 140.
The composition according to any one of claims 1 to 3, which is obtained by hot water extraction under a temperature condition of ° C, preferably 90 to 125 ° C. 5. The composition according to any one of claims 1 to 4, wherein the solvent extract is obtained by extracting the cultured mycelium with methanol. 6. The solvent extract is the solvent extract itself, a liquid part obtained by removing the extraction residue therefrom, a concentrate thereof, a paste, a diluent, and / or a dried product. The composition according to any one of claims 1 to 5, characterized in. 7. The composition according to claim 1, wherein the composition is a pharmaceutical type and / or a food / beverage type. 8. A pharmaceutical type composition according to claim 7, characterized in that said composition is a cardiotonic or antitussive. 9. The food-drink type composition according to claim 7, wherein the composition is a health drink.