KR20090036674A - Mushroom Extract for Treating Gastritis and Gastric Ulcer Disease - Google Patents

Abstract

Mushroom extracts for treating gastric diseases is provided to prevent stomach disorder by cultivating Hericium erinaceum mycelium extract or Reishi mushroom mycelium extract cultivated in an Artemisia capillaris thunb culture medium. Mushroom extracts for treating gastric diseases is Hericium erinaceum mycelium extract or Reishi mushroom mycelium extract cultivated in an Artemisia capillaris thunb culture medium. The mycelium extract comprises at least one compound of eicosane, 5-eicosene, eicosanol, eicosanoic acid ethyl ester, hexadecancic acid, hexadecanoic acid ethyl ester, 1-octadecanol, octadecanoic acid ethyl ester, 9-octadecenoic acid ethyl ester, 9,12-octadecadienoic acid ethyl ester, ethyl tetracosanoate, phytol, linolenic acid ethyl ester, eicosapentanoic acid, eicosapentanoic acid ethyl ester, hexadecanoic acid, docosahexaenoic acid, di-octyl phthalate and 6,7-dimethoxycoumarin.

Description

Mushroom Extracts for Treating Gastritis and Gastric Ulcer Disease

 The present invention relates to a mushroom extract useful for treating gastrointestinal diseases such as gastritis and gastric ulcer. More specifically, the present invention relates to a gastrointestinal disease treatment agent such as gastritis and gastric ulcer, which contains as an active ingredient, extracts of the Roestock fungus mycelium cultured in Injin mugwort medium and Ganoderma lucidum mycelium extract cultured in Injin mugwort medium.

The pharmacological activity of mushroom extracts or mushroom cultures is already well known. For example, it has been reported that extracts of Roeden mushroom culture can protect liver cancer, hepatitis and other liver damage by inhibiting the formation of 8,9-epoxy aflatoxin B ₁ (Food Sci. Biotechnol .. Vol. 12 , No.2, pp 183-186, (2002), Republic of Korea Patent Publication No: 10-2005-0039390, J. Ethnopharmacology, 100, pp 176-179, (2005)), also caused by the hyperproliferation of smooth muscle cells It is known that the disease can be prevented and treated (Patent Publication No. 10-2005-0079788). Also, a method for culturing roe deer mushrooms or ganoderma lucidum in a solid medium containing phosphorus wormwood is known in the art (public disclosure). Patent Publication 2005-0012054) The mushroom extract cultured by the above method is used as a food additive for health promotion. It is known that the locust mushroom extract cultured in a solid medium containing phosphorus wormwood is effective in relieving hangover by promoting the decomposition of acetyl aldehyde and promoting the decomposition of alcohol, and can be usefully used as a food additive. No. 2005-0079790) In addition, it has been shown that the extract of the locust mushroom culture has antimicrobial activity against Helicobbacter pylori, which has been found to cause gastritis and gastric ulcers. ).

Helicium erinaceum belongs to the Aphyllophorales Beard Mushroom (Hydnaceae) and has high beta-D-glucan content and strong superoxide dismutase ability. It is also in the spotlight as a health food. It is known that 100g of scab mushroom contains about 20-30g of beta-D-glucan (glycan polysaccharide), which inhibits the proliferation of cancer cells and has a high anti-cancer effect.Indigestion, nervous breakdown, physical weakness, nourishment It is effective in preventing recurrence of tonic, gastrointestinal cancer, and surgery. It is used as an antioxidant to prevent the oxidation of cells and prevent aging and carcinogenesis. It is known to have an effect of improving immune function and biological function by enhancing immunity.

Artermisia capillaris is called wormwood and wormwood in Korea. Injin mugwort is a health food containing capillene, pinene, and vitamin B _1, and is known to have an excellent effect on the liver. According to a recent study, the ethanol fraction of the extract fraction of Artermisia asiatica, a stem of Injin mugwort, was isolated from the ethanol fraction, and the isolated substance was eupatilin, a flavonoid compound. Confirmed. In addition, some mechanisms of action have been studied, such as anti-ulcer effects on mucus extracts, effects on mucus secretion, effects on acids and acid secretion by stimulation (The Journal of Applied Pharmacology, 4, 111-121). (1996)). Extracted from 70 to 100% high concentration of ethanol extracts from jinseng leaves, contains no water-soluble components that show the side effects of blood coagulation inhibition, contains high concentrations of eufatlin and jaceosidin, and inhibits gastric ulcer lesions and protects the cells. Mugwort extracts have been reported to simultaneously exhibit prostaglandin biosynthesis promoting activity (published patent No. 1996-0021054). Also, it was found to be safe in general pharmacology, acute toxicity and toxicity evaluation by 4 weeks of repeated administration. , (The Journal of Applied Pharmacology, 4, 354-363 (1996), The Journal of Applied Pharmacology, 4, 174-183 (1996)) These extracts are also used to treat inflammatory bowel diseases such as gastritis, ulcerative colitis and Crohn's disease. (Patent Publication No. 10-1998-0008235, Dong-A Pharmaceutical, Trade Name: Styrene)

H ₂ in gastric ulcer and gastrointestinal disease therapeutic agent-receptor antagonist cimetidine (cimetidine, Ranitidine), LA motif Dean (Ramotidine) etc. have been developed, H ⁺ - pump inhibitors Omeprazole (Omeprazole), misoprostol (Misoprostol), etc. Prostaglandin-based drugs include Alprostadil, Dinoprostone, Dinoprost, and Carbonprost. Phosphorus mugwort extract is known to show prostaglandin biosynthesis promoting activity (The Journal of Applied Pharmacology, 4, 111-121 (1996)).

It is already known that the extracts of Injin mugwort, Roe deer fungus mycelium, and Ganoderma lucidum mycelium extract exhibit physiological activity in gastrointestinal diseases. The present inventors found that the conventional extracts did not achieve a satisfactory result of inhibiting gastric lesions as a gastrointestinal disease treatment, and the mushroom extracts of the present invention were in search of ways to increase their gastric lesion suppression rate. Comparing to the marketed gastrointestinal therapeutics consisting of the extract of the Artemisia mugwort, Roe deer fungus mycelium extract or Ganoderma lucidum mycelium extract, it was confirmed that the present invention exhibits excellent gastric lesion suppression efficacy, and completed the present invention.

The present invention is to provide a new use by confirming that the extracts of Roeden beetle mycelium mycelium cultured in Injin mugwort medium or the Ganoderma lucidum mycelium extract cultured in Injin mugwort medium shows excellent effects in the treatment of gastritis and gastric ulcer disease.

It is an object of the present invention to provide an extract of the mycelial fungus mycelium cultured in Injin mugwort medium and Ganoderma lucidum mycelium cultured in Injin mugwort medium useful for treating gastrointestinal diseases such as gastritis and gastric ulcer.

The present invention relates to the structural analogues of the extract of Rhizome myrtle mycelium cultured in Injin mugwort medium, Ganoderma lucidum mycelium cultured in Injin mugwort medium, and compounds showing physiological activity, which are useful for the treatment of gastrointestinal diseases such as gastritis and gastric ulcer. will be.

The mushroom extract of the present invention can be obtained by extracting the locust mushroom mycelium and Ganoderma lucidum mycelium cultured in Injin mugwort medium with an organic solvent such as water, alcohol, n-hexane, chloroform, methylene chloride, ethyl acetate, butanol.

The extract of Mycelia of the Roe beetle mushrooms includes eicosane, 5-eicosene, 5-eicosene, eicosanol, eicosanoic acid ethyl ester, and hexadecanoic acid. ), Hexadecanoic acid ethyl ester, 1-octatecanol, 1-octadecanol, octadecanoic acid ethyl ester, 9-octadecenoic acid ethyl ester (9-octadecenoic acid ethyl ester), 9,12-octadecadienoic acid ethyl ester (9,12-octadecadienoic acid ethyl ester), ethyl tetracosanoate, phytol, linolenic acid ethyl Linolenic acid ethyl ester, eicosapentanoic acid, eicosapentanoic acid ethyl ester, docosa hexaenoic acid, docosahexaenoic acid One or more compounds selected from the group consisting of di-octyl phthalate and 6,7-dimethoxycoumarin, which are not included in mushroom extracts using the Roots of the Artemisia medium, but are active It includes analogous structural compounds of the substances that represent.

Hereinafter, an Example is given and this invention is demonstrated concretely.

< Injin mugwort  On the badge Roe deer  Cultivation of Mushroom Mycelium>

After mixing 1 kg of dried phosphorus mugwort and 2 liters of water, the solid medium obtained by sterilization at 110-130 ° C. for 2 to 4 hours was inoculated with 10% by weight of the medium weight of the snail mushroom mycelium cultured in YMPG medium and then 23 ~. The mycelia are grown at 26 ° C. for 40-50 days.

After the primary growth, the solid medium in which the snail mushroom mycelium was formed was dried before the fruiting body was formed, thereby obtaining the snail mushroom mycelium culture cultured in the jinjin mugwort medium.

In this process, MCM medium may be used instead of YMPG medium.

< Injin mugwort  Cultivation of Ganoderma lucidum Mycelium in Medium>

A Ganoderma lucidum mycelium culture is obtained in the same manner as the method of cultivating the locust mushroom mycelium in the Injin mugwort medium, except that the Ganoderma lucidum mushroom is used instead of the locust mushroom.

 Mushroom mycelium cultivation method for obtaining the roe deer mushroom culture in Injingumi medium is already known in the art (Publication Publication No. 05-0012054).

<Preparation of Roebuck Mushroom Mycelium Extract Cultured in Injinguru Sacrum Medium>

2L of anhydrous ethanol was added to 1 Kg of the dried Artemisia agar medium containing 1B of the Roe mushroom mycelium. After immersion for 3-4 days, the mixture was filtered and concentrated to obtain 34.8g of an extract (extract A ₁ ) extracted with anhydrous ethanol. In addition, 2L of 70% ethanol (v / v) was added to 1 Kg of dried wormwood medium containing 1M ginseng mycelium mycelium. After immersion for 3-4 days, the mixture was filtered and concentrated to obtain 120.75 g of the extract (extract A ₂ ) extracted with 70% ethanol. In addition, 2L of 50% ethanol (v / v) was added to 1 Kg of dried wormwood medium containing the Roe mushroom mycelium mycelium. After immersion for 3-4 days, the mixture was filtered and concentrated to obtain 162.3 g of the extract (extract A ₃ ) extracted with 50% ethanol. , 30% ethanol (v / v) 2L was added to 1Kg of dried jinjinji medium containing 1Gg of mycelium fungus mycelium, immersed for 3-4 days, filtered and concentrated to obtain 221.6g of extract (extract A ₄ ) extracted with 30% ethanol. 2L of water was added to 1 Kg of dried jinjinjinji medium containing roe deer fungus mycelium, immersed for 3-4 days, filtered and concentrated to obtain 260g of the extract (extract A ₅ ) extracted with water.

< Injin mugwort  Preparation of Ganoderma lucidum Mycelium Extract Cultured in Medium>

2L of anhydrous ethanol was added to 1 Kg of dried Artemisia agar medium containing the Ganoderma lucidum mycelium. After immersion for 3-4 days, the mixture was filtered and concentrated to obtain 36.8 g of an extract (extract B ₁ ) extracted with anhydrous ethanol. In addition, 2L of 70% ethanol (v / v) was added to 1 Kg of dried jinjinjiji medium containing Ganoderma lucidum mycelium. After immersion for 3-4 days, the mixture was filtered and concentrated to obtain 112.5g of the extract (extract B ₂ ) extracted with 70% ethanol. In addition, 2L of 50% ethanol (v / v) was added to 1 Kg of dried jinjinjiji medium containing Ganoderma lucidum mycelium. After immersion for 3-4 days, the mixture was filtered and concentrated to obtain 142.6 g of the extract (extract B ₃ ) extracted with 50% ethanol. 2L of 30% ethanol (v / v) was added to 1 Kg of dried jinjinjiji medium containing Ganoderma lucidum mycelium, immersed for 3-4 days, filtered and concentrated to obtain 225.4 g of an extract extracted with 30% ethanol (extract B ₄ ). 2L of water was added to 1 Kg of dried jinjinjiji medium containing Ganoderma lucidum mycelium, immersed for 3-4 days, filtered and concentrated to obtain 184g of the extract (extract B ₅ ) extracted with water.

Preparation of Ganoderma lucidum Extract

2L of anhydrous ethanol was added to 1Kg of Ganoderma lucidum mushroom dried, immersed for 3-4 days, filtered and concentrated to obtain 14.8g of an extract (extract C ₁ ) extracted with anhydrous ethanol. In addition, 2L of 70% ethanol (v / v) was added to 1Kg of dried Ganoderma lucidum, immersed for 3-4 days, filtered, and concentrated to obtain 23.6 g of an extract (extract C ₂ ) extracted with 70% ethanol. In addition, 2L of 50% ethanol (v / v) was added to 1Kg of dried ganoderma lucidum, which was immersed for 3-4 days, filtered and concentrated to obtain 28.3 g of an extract (extract C ₃ ) extracted with 50% ethanol, and to 1Kg of ganoderma lucidum dried 2L of 30% ethanol (v / v) was added, immersed for 3-4 days, filtered, and concentrated to obtain 27.6 g of an extract (extract C ₄ ) extracted with 30% ethanol. 2L of water was added to 1Kg of dried ganoderma lucidum, immersed for 3-4 days, filtered and concentrated to obtain 18.4 g of an extract (extract C ₅ ) extracted with water.

< Roe deer  Preparation of Mushroom Extracts>

2L of anhydrous ethanol was added to 1 Kg of dried deer worm mushroom, and after immersion for 3-4 days, the mixture was filtered and concentrated to obtain 17.5 g of an extract (extract D ₁ ) extracted with anhydrous ethanol. In addition, 2L of 70% ethanol (v / v) was added to 1 Kg of dried deer worm fungi, filtered and concentrated for 3-4 days to obtain 26.3 g of an extract (extract D ₂ ) extracted with 70% ethanol. In addition, 2L of 50% ethanol (v / v) was added to 1 Kg of dried deer worm mushroom mushrooms, immersed for 3-4 days, filtered, and concentrated to obtain 32.4 g of an extract (extract D ₃ ) extracted with 50% ethanol. 2L of 30% ethanol (v / v) was added to 1 Kg, immersed for 3-4 days, filtered, and concentrated to obtain 28.3 g of an extract extracted with 30% ethanol (extract D ₄ ). 2L of water was added to 1 Kg of dried deer worm mushroom, immersed for 3-4 days, filtered, and concentrated to obtain 11.7 g of an extract (extract D ₅ ) extracted with water.

< Injin mugwort  Preparation of Extracts>

2L of anhydrous ethanol was added to 1 Kg of dried Artemisia jinju, filtered and concentrated for 3-4 days to obtain 36.4 g of an extract (extract E ₁ ) extracted with anhydrous ethanol. In addition, 2L of 70% ethanol (v / v) was added to 1 Kg of dried Mugwort, filtered and concentrated for 3-4 days to obtain 89.7 g of an extract (extract E ₂ ) extracted with 70% ethanol. In addition, 2L of 50% ethanol (v / v) was added to ₁ Kg of dried mugwort, filtered and concentrated for 3-4 days to obtain 134.6 g of the extract (extract E ₃ ) extracted with 50% ethanol, 30% to 1 kg of dried mugwort. 2L of ethanol (v / v) was added, immersed for 3-4 days, filtered and concentrated to obtain 142.7 g of an extract (extract E ₄ ) extracted with 30% ethanol. 2L of water was added to 1 Kg of dried Mugwort, immersed for 3-4 days, filtered, and concentrated to obtain 235.7 g of an extract (extract E ₅ ) extracted with water.

The extracts from the mycelia of the Roestock fungus Mycelium and the Ganoderma lucidum mycelium cultured in Injin mugwort medium have higher inhibitory effect on gastric lesions than those of the extracts of the Rosacea fungus, Ganoderma lucidum extract and Root of the Artemisia.

Example  One.

<Test of Gastric Lesion Inhibitory Effect of Mushroom and Injin mugwort Extract>

Gastric lesion suppression experiments with extracts were administered by oral administration of a 7-week-old Sprague-Dawley male for 24 hours after fasting. Oral drug is prepared by suspending the amount of each extract 70 mg / Kg in 5 ml of distilled water. One hour after the administration of the drug, 1.0 mL / head of gastric lesion-producing reagent 150 mM HCl-EtOH (60%) solution is administered orally. After 3 hours, the rats are killed and the stomach is removed. After extraction, 10 mL of 4% formalin is injected into the stomach and fixed in this solution for 1 hour. Incision and washing along the Taiwan part in the water flowing through the fixed stomach to measure the degree of ulcers. Gastric disease inhibition experiment according to the fraction was carried out in the same manner as the concentration of the hexane layer, chloroform layer, ethyl acetate, butanol and water layer as a dosage drug, and also purified and analyzed by GC / MS and chromatography Gastric lesion suppression experiment was performed in the same way. Gastric lesion inhibition rate was calculated by the following equation generally adopted in this field, and the significance test with the control group was Student's t-test, One-way Analysis of Variacnce (ANOVA) and Dunnett's Test.

                   Gastric injury area of control muscle-Gastric injury index of drug group

Gastric lesion inhibition rate (%) = ------------------------------------------- ------ × 100

                             Gastric impairment index of the control group

Table 1. Gastric lesion inhibition rate according to the extract

extract Number of test animals % Inhibition 100% Ethanol Extract (A ₁ ) 10 69.0 70% Ethanol Extract (A ₂ ) 10 36.0 50% Ethanol Extract (A ₃ ) 10 27.0 30% Ethanol Extract (A ₄ ) 10 40.0 Water extracts (A ₅ ) 10 40.0 100% Ethanol Extract (B ₁ ) 10 38.0 70% Ethanol Extract (B ₂ ) 10 24.0 50% Ethanol Extract (B ₃ ) 10 22.0 30% Ethanol Extract (B ₄ ) 10 41.0 Water extract (B ₅ ) 10 28.0 100% Ethanol Extract (C ₁ ) 10 41.0 70% Ethanol Extract (C ₂ ) 10 30.0 50% Ethanol Extract (C ₃ ) 10 26.0 30% Ethanol Extract (C ₄ ) 10 25.0 Water Extract (C ₅ ) 10 26.0 100% Ethanol Extract (D ₁ ) 10 7.0 70% Ethanol Extract (D ₂ ) 10 8.0 50% Ethanol Extract (D ₃ ) 10 6.0 30% Ethanol Extract (D ₄ ) 10 9.0 Water extract (D ₅ ) 10 6.0 100% Ethanol Extract (E ₁ ) 10 8.0 70% Ethanol Extract (E ₂ ) 10 9.0 50% Ethanol Extract (E ₃ ) 10 7.0 30% Ethanol Extract (E ₄ ) 10 5.0 Water extract (E ₅ ) 10 8.0

A ₁ ~ A ₅ : Injin mugwort extract containing the mushrooms

B ₁ ~ B ₅ : Injin mugwort extract containing Ganoderma lucidum mycelium

C ₁ C ₅ : Mugwort Extract

D ₁ ~ D _5: Hericium erinaceus extract

E ₁ ~ E ₅ : Ganoderma lucidum extract

Example 2.

<Test of Gastric Lesion Inhibitory Effect of Mushroom Mycelia Extract Solvent Extract Fraction>

To 30 g of 100% ethanol extract (Extract A ₁ ) of the Roe mushroom fungus mycelium cultured in Injingumi medium, 200 ml of water was added and slurried for 30 minutes, 200 ml of hexane was added thereto, followed by stirring to separate the hexane layer and the water layer. The hexane layer was separated, and hexane was removed therefrom and concentrated to obtain 0.94 g of a hexane fraction (fraction F ₁ ).

200 ml of chloroform was added to the separated water layer, and the mixture was stirred to separate the layers into a chloroform layer and a water layer. The chloroform layer was separated, chloroform removed, and concentrated to give 12.3 g of a chloroform fraction (fraction F ₂ ).

200 ml of ethyl acetate was added to the separated water layer, and the mixture was stirred to separate the layers into an ethyl acetate layer and a water layer. The ethyl acetate layer was separated, and ethyl acetate was removed and concentrated to obtain 4.3 g of ethyl acetate fraction (fraction F ₃ ). 200 mL of butanol was added to the separated water layer and stirred to separate the butanol layer and the water layer. The butanol layer was separated and the butanol was removed and concentrated to obtain 3.4 g of butanol fraction (fraction F ₄ ).

Water was finally removed from the separated water layer and concentrated to give 7.2 g of water fraction (Fragment F5). Inhibition rate of gastric lesions of gastritis treatment (Trebex, Hanil Pharm.) Based on teprenone (trade name: Styrene, Dong-A Pharmaceutical Co., Ltd.) Was tested and the results are shown in Table 2 below.

As shown in Table 2 below, the fractions obtained in the present invention show superior gastric lesion inhibition rate compared to conventional gastritis treatment agents.

Table 2. Inhibition of Gastric Lesions by Solvent Fraction Extract

Reference Drugs and Fraction Extracts Number of test animals % Inhibition Styrene 10 52.7 Selves 10 53.8 Hexane fraction (fraction F ₁ ) 10 87.0 Chloroform fraction (fraction F ₂ ) 10 89.0 Ethyl acetate fraction (fraction F ₃ ) 10 25.4 Butanol fraction (fraction F ₄ ) 10 27.5 Water layer fraction (fraction F ₅ ) 10 31.5

Example  3.

<Compositional Components Test of Extracts of Roebuck Mushroom Mycelium Cultured in Injinguru Saccharic Medium>

Figure 1 is a high performance liquid chromatograph (hereinafter referred to as HPLC) of the extract of the Roestock fungi (HPLC), Figure 2 is the HPLC of the fungus mugwort extract, Figure 3 is the HPLC of the extract of the mycelia of the fungus incubated in the intake media. As shown in FIGS. 1-3, the three HPLC chromatograms are different, and in particular, the different peaks appearing in FIGS. 2 and 3 indicate that certain components of the jinjin mugwort are changed to other components during the culture. In other words, it is determined that some of the components of the ginseng mugwort are changed to other ingredients during the cultivation of the Roe beetle mushroom hyphae using the ginseng mugwort as a medium. In addition, in Table 2, the GC chromatogram of the physiologically active fraction of chloroform is shown in Fig. 4 and consists of several components. The structure of these components was confirmed by GC / MS, and the results of GC / MS analysis predicted the eicosane compound of the peak at 31.01 minutes of GC. As a result, it was confirmed that the compound of this peak was eicosane. 5-eicosene at 25.1 minutes, eicosanol at 35.1 minutes, eicosanoic acid ethyl ester at 23.5 minutes, and hexadecanoic acid at 16.1 minutes. ), Hexadecanoic acid ethyl ester at 16.6 minutes, 1-octadecanol at 28.1 minutes, octadecanoic acid ethyl ester at 26.7 minutes, 19.8 9-octadecenoic acid ethyl ester in minutes, 9,12-octadecadienoic acid ethyl ester in 19.3 minutes, ethyl tetra in 29.5 minutes Ethyl tetracosanoate, phytol at 18.9 minutes, linolenic acid ethyl ester at 19.5 minutes, di-octyl phthalate at 46.2 minutes, 6,7 at 36.5 minutes -Dimethoxy comarin (6,7-dimethoxycoumarin) was found to be. HPLC conditions of FIGS. 1-3 and GC / MS conditions of FIGS. 4 and 5 are as follows.

* HPLC conditions

Mobile phase: Acetonitrile: Buffer = 4: 6

              (Buffer solution: 2.5% acetic acid, 5% ammonium acetate)

Flow rate: 0.5 ml / min

UV detection: 360 nm

Column: Capcell pak C18 (4.6 mm × 250 mm 5μm)

* GC / MS condition

Column: HP-10

Carrier gas: Helium

Flow rate: 1.5 ml / min

Injector temp. : 280 ℃

Oven temp. : start at 120 ℃

             stand for 5min

             increase 10 ℃ / min

             stand for 5min

Terminal auxl temp. : 300 ℃

Dectector voltage: 1.3kV

Library: NIST. shimadzu Co.

As shown in Table 1, it can be seen that the gastrointestinal ulcer fungus culture cultured in jinjin mugwort medium has a very good gastritis ulcer inhibitory effect than the jinja extract. In addition, the effect of the extraction method was found to be very excellent when extracted with pure ethanol extract. Water was added to the pure ethanol extract and extracted sequentially with hexane, chloroform, ethyl acetate and butanol, respectively, to obtain a concentrated solution of each solvent. Each concentrated solution was tested according to Injin mugwort extract (trade name: Dong-A Pharmaceutical Styrene), Tefrenone (Teprenone) gastritis treatment agent (trade name: Selbex, Hanil Pharmaceutical Products) and Example 2 are shown in Table 2. As shown in Table 2, hexane extracts and chloroform extracts of Injin mugwort cultures showed much higher therapeutic effects than comparable drugs. In order to determine the main component of the chloroform concentrate, which was the most effective, it was analyzed using gaschromatography / mass spectrometer (GC / MS). 4 and 5 show the results of GC chromatograms in which the extract of Injin mugwort was extracted with ethanol and extracted with chloroform in the same manner. It can be seen that the extracts of Injin mugwort culture are different. These results indicate that new substances are produced during the incubation of mycelial and roe deer fungus hyphae, and these materials have a superior therapeutic effect on gastritis and gastric ulcers.

GC / MS analysis showed that the mass spectrum of the peak at 31.01 minutes of GC predicts that it is an eicosane compound and compares it with the standard eicosane spectrum. It confirmed that it was eicosane. In addition, as a result of the mesospectral analysis of the 25.1 minute peak of GC, it was confirmed that it was 5-eicosene. In the same manner, the 33.8 minute peak of GC was eicosanol, and the eicosanoic acid ethyl ester at 23.5 minutes. eicosanoic acid ethyl ester, hexadecanoic acid at 16.1 minutes, hexadecanoic acid ethyl ester at 16.6 minutes, 1-octadecanol at 28.1 minutes, 26.7 minutes Octadecanoic acid ethyl ester, 9-octadecenoic acid ethyl ester at 19.8 minutes, 9,12-octadecadinoic acid ethyl ester at 19.3 minutes 9,12-octadecadienoic acid ethyl ester, ethyl tetracosanoate at 29.5 minutes, phytol at 18.9 minutes, linolenic acid ethyl ester at 19.5 minutes, diethyl at 46.2 minutes Octyl phthalate and 36.5 minutes 6, 7-dimethoxy-coumarin was found to be (6,7-dimethoxycoumarin). On the other hand, using silica gel column chromatography, among them, eicosane, 5-eicosene, eicosanol, eicosanoic acid ethyl ester, hexadecanoic acid acid, hexadecanoic acid ethyl ester, 1-octadecanol, octade Canoic acid ethyl ester, 9-octadecenoic acid ethyl ester, 9,12-octadecadienoic acid ethyl ester, ethyl tetracosanoate, phytol, linolenic acid ethyl ester, dioctylphthalate and 6,7- Dimethoxy commarin was purified and purified by pure GC / MS, IR, and H ¹ -NMR spectra to confirm the structure was consistent with the results of the GC / MS analysis.

Among the components, the active material was identified to have a fatty acid structure containing 1 to 3 single bonds or double bonds or a fatty acid ethyl or methyl ester compound and a long chain alcohol group. Table 3 shows the results of the treatment of gastritis and gastric ulcer on the main components of chloroform concentrate. As shown in Table 3, most compounds showed therapeutic effect, but among them, ester compounds of fatty acids, octadecanoic acid ethyl ester, 9-octadecenoic acid ethyl ester, and 9,12-octadecadinoic acid ethyl Ester, ethyl tetracosanoate, phytol, linolenic acid ethyl ester, dioctylphthalate and 6,7-dimethoxycoumarin showed excellent therapeutic effects. In addition, it was confirmed that the mixture of these compounds has a higher gastric lesion therapeutic effect.

Table 3. Inhibition rate of main components showing physiological activity

Compound name Number of test animals % Inhibition Styrene 10 52.3 Selves 10 53.5 Eicosanol (A) 10 36.3 1-octadecanol (B) 10 45.2 Phytol (C) 10 53.9 Hexadecanoic acid (D) 10 50.9 Hexadecanoic acid ethyl ester (E) 10 88.3 Octadecanoic Acid Ethyl Ester (F) 10 85.2 9-octadecenoic acid ethyl ester (G) 10 90.3 9,12-octadecadienoic acid ethyl ester (H) 10 92.5 Linolenic acid ethyl ester (J) 10 84.5 6,7-dimethoxycoumarin (6) 10 95.0 Dioctylphthalate (L) 10 62.3 E + G 10 88.2 E + H 10 88.9 G + H 10 90.6 E + G + H 10 93.3

Styrene: Dong-A Pharm Injin mugwort extract drug

Selbex: Brand name of Hanil Pharm. Inc.

Example  4.

Looking at the component structure of Example 3 it was confirmed that most of the unsaturated fatty acid or unsaturated fatty acid ester. Therefore, E. coli extracts from the mycelia of the Roots of the Roots of the Roots of the Roots of the Roots, but similar structures eicosaptananoic acid, eicosapentanoic acid ethyl ester and docosa hexaenoic acid, etc. Was purchased and tested in the same manner as in Example 1. The results are shown in Table 4.

Table 4. Inhibition Rates of Active Compound-like Structural Compounds

Compound name Number of test animals % Inhibition Eicosapentanoic acid 10 72.3 Eicosapentanoic acid ethyl ester 10 73.5 Docosa hexaenoic acid (docosahexaenoic acid) 10 72.8

Example  5.

Administration of the extract of the present invention, 100% ethanol extract, chloroform fractionation layer, 9,12-octadecadinoic acid ethyl ester and 6,7-dimethoxycomarin which showed the highest physiological activity among the solvent fraction layer and the single substance In order to confirm the dose dependency by changing the amount to 150, 75, 65, 60, 50, 40, 30, 20 and 10 mg / Kg, the inhibition rate according to the dose was confirmed in the same manner as in Example 1, and the dose-dependent curve graph The ED ₅₀ values, which are doses representing 50% inhibition rate, are obtained from Table 5 below.

Table 5. ED ₅₀ values of extracts and monomaterials

A ₃ : 50% ethanol extract of Injin mugwort broth containing roe deer fungus mycelium

F ₂ : Chloroform Fraction of Injin mugwort containing Roebuck Mushroom Mycelia

H: 9,12-octadecadinoic acid ethyl ester

K: 6,7-dimethoxycomarin

Figure 1 is a high-performance liquid chromatograph of roe deer mushroom extract.

2 is a high-performance liquid chromatograph of Injin mugwort extract.

Figure 3 is a high-speed chromatograph of the extract of the mycelia of the Roe beetle cultured in Injinguru medium.

Figure 4 is a gas chromatograph of the extract of the hemperae mushroom cultured in jinjinmugi medium.

5 is a gas chromatograph of the extract of Injin mugwort.

Claims (3)

A mycelium mycelia extract cultured in Injin mugwort medium for the treatment of gastritis and gastric ulcer disease. Extract of Ganoderma lucidum mycelium cultured in Injin mugwort medium for the treatment of gastritis and gastric ulcer disease. The method according to any one of claims 1 to 3, wherein eicosane, 5-eicosene, eicosanol, eicosanoic acid ethyl ester, hexa Hexadecanoic acid, hexadecanoic acid ethyl ester, 1-octatecanol, octadecanoic acid ethyl ester, 9- 9-octadecenoic acid ethyl ester, 9,12-octadecadienoic acid ethyl ester, ethyl tetracosanoate, phytol phytol, linolenic acid ethyl ester, eicosapentanoic acid, eicosapentanoic acid ethyl ester, docosa hexaenoic acid , Di Butyl phthalate (di-octyl phthalate), and 6,7-dimethoxy-coumarin mushroom mycelium extract containing at least one compound selected from the group consisting of (6,7-dimethoxycoumarin).

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