KR20220157172A - A Composition For Preventing, Improving Or Treating Gastrointestinal Disorders Including Extract Of Eucommia Ulmoides Oliver And Extract Of Citrus Unshiu - Google Patents

Abstract

The present invention relates to a composition for the prevention, alleviation, or treatment of gastrointestinal disorders, which contains an Eucommia ulmoides extract and a Citrus unshiu extract as active components, thereby having an effect of suppressing excessive secretion of gastric acid and alleviating stomach damage, and thus being used to prevent, alleviate or treat gastrointestinal disorders by drinking frequently in daily life since extracts derived from natural substances are used as active ingredients, and toxicity is low.

Description

A composition for preventing, improving or treating gastrointestinal disorders comprising Eucommia extract and dermis extract {A Composition For Preventing, Improving Or Treating Gastrointestinal Disorders Including Extract Of Eucommia Ulmoides Oliver And Extract Of Citrus Unshiu}

The present invention relates to a composition for preventing, improving or treating gastrointestinal disorders comprising an extract of Eucommia and a dermis extract.

Gastric hyperacidity is a symptom of excessive secretion of stomach acid in the stomach, which is a common gastrointestinal disorder that can occur even in healthy people. It can also occur in the case of chronic gastrointestinal disease, but it can also occur temporarily when life rhythms such as mental anxiety, stress, and irregular meals are disrupted. Symptoms of gastric hyperacidity include epigastric pain, pain on an empty stomach (the symptoms disappear temporarily when food is eaten), stomach ache, belching, sourness, rumination, and bowel movements. When these symptoms accumulate, gastritis and peptic ulcer , can progress to various diseases such as reflux esophagitis.

Peptic ulcer (Peptic ulcer) commonly means gastric ulcer or duodenal ulcer, and refers to a condition in which the mucous membrane of the stomach or duodenum is damaged or lost for some reason and submucosal tissue is exposed. When the lesion is localized only to the mucous membrane, it is called erosion, and the word gastritis or duodenitis is also used because the stomach or duodenum is usually broken. Peptic ulcer is a common disease that occurs in 5-10% of the population worldwide, and it is known that men are about 2 to 3 times more likely than women. Duodenal ulcers often occur at a younger age than gastric ulcers.

For the treatment of gastrointestinal disorders, antacids and histamine receptor antagonists are mainly used, but even after gastritis or peptic ulcer is cured, it often recurs or reproduces, so long-term maintenance therapy is required even after complete healing. Accordingly, studies on a composition that can prevent or improve gastrointestinal disorders by suppressing excessive secretion of gastric acid and that can be consumed on a daily basis have been continuously conducted.

Eucommia ulmoides Oliver ) represents a deciduous tree belonging to the Eucommia family. It is 10m high, the bark is brownish grayish white, and many branches are split. Leaves are alternate, 5-16cm long, 2-7cm wide, elliptical, and petiole is 1cm long. Flowers are unisexual, blooming in the leaf axils in April, male flowers have 6-10 short stamens, reddish brown, and female flowers have one short stalk. The fruit is a sap and is oblong in shape of 3-3.5cm in length in October-November. Eucommia stems, leaves, etc. have been drunk as tea since ancient times, blood pressure lowering action, diuretic action, etc. are known, and the stems, leaves, etc. are known to be useful for various diseases such as stress and hangover.

The dermis refers to the mature rind of a tangerine ( Citrus unshiu Markovich ) or a related plant of the same genus. Appearance is an irregular shell, yellowish red or dark tan on the outer surface, white or light grayish brown on the inside, light and brittle. The dermis relieves qi and strengthens the function of the spleen, treating symptoms such as belching, vomiting, nausea, indigestion, flatulence and drowsiness, and loose stools. It removes seawater and phlegm and acts as a diuretic. It is a medicine whose medicinal effect increases with age along with recitative, mahuang, dermis, ohsuyu, banha, and jisil. It has been reported that essential oils stimulate digestion, promote digestion, expectorant, anti-ulcer, cardiac, increase blood pressure, anti-allergy, promote bile secretion, inhibit uterine smooth muscle, and antibacterial.

The present inventors studied whether materials derived from natural products can be used to improve gastrointestinal disorders, and among natural products, when dermal extract and Eucommia extract are mixed in a specific ratio, prevention of gastrointestinal disorders by inhibiting excessive secretion of gastric acid, mitigating the degree of gastric damage, It was confirmed that the improvement or treatment effect was excellent, and the present invention was completed.

Korean Patent Publication No. 10-2009-0046425 (2009.05.11.)

An object of the present invention is to provide a composition effective for preventing, improving or treating gastrointestinal disorders.

One aspect of the present invention provides a pharmaceutical composition for preventing or treating gastrointestinal disorders comprising an extract of Eucommia and a dermis extract.

Another aspect of the present invention provides a health functional food composition for preventing or improving gastrointestinal disorders, including Eucommia extract and dermal extract.

The composition for preventing, improving, or treating gastrointestinal disorders of the present invention is excellent in inhibiting hypersecretion of gastric acid and alleviating the degree of gastric damage by including natural substances, Eucommia extract and dermis extract, in a specific weight ratio. In addition, since the composition for preventing, improving or treating gastrointestinal disorders of the present invention contains an extract derived from a natural substance, it has low toxicity and can be consumed at any time on a daily basis, thereby effectively preventing, improving or treating gastrointestinal disorders. have.

Figure 1 shows the result of confirming the cAMP level (%) of the experimental group compared to the control group.
Figure 2 shows the results of confirming the H + / K + ATPase activity (%) of the experimental group compared to the control group.
Figure 3 shows the result of confirming the gastric juice secretion (%) of the experimental group compared to the control group.
Figure 4 shows the results of confirming the gastric wall damage index (%) of the experimental group compared to the control group.
5 shows the result of confirming the blood histamine concentration (%) of the experimental group compared to the control group.

Hereinafter, the present invention will be described in more detail to aid understanding of the present invention. At this time, the terms or words used in this specification and claims should not be construed as being limited to ordinary or dictionary meanings, and the inventor appropriately defines the concept of terms in order to explain his/her invention in the best way. It should be interpreted as meaning and concept consistent with the technical idea of the present invention based on the principle that it can be done.

1. Preparation of Eucommia extract and dermal extract

In the present invention, Eucommia is a leaf, young sprout, body, body shell, stem, stem shell, root, root shell, rhizome, seed, fruit, immature fruit, mature fruit, fruit flesh, fruit skin, flower, stamen group (androecium), Any one selected from the group consisting of a pistil group (gynoecium), calyx, stamen, petal, calyx piece, carpel, and combinations thereof may be used, and preferably stems or leaves are used.

In the present invention, the dermis may be at least one selected from the group consisting of fruit, leaf, flower, bark, and root of a Citrus unshiu Markovich tree, preferably a fruit. The dermis may use any one selected from the group consisting of flesh, skin, seeds, and combinations thereof, and preferably use skin.

In the present invention, 'extract' refers to a material extracted from a raw material by any method, and is used in the sense of including all of the extracted extract, the concentrate obtained therefrom, and the dried product and powder of the concentrate without limitation.

When the extract is obtained by extraction from a raw material, all known conventional extraction methods such as solvent extraction, ultrasonic extraction, filtration and reflux extraction may be used as extraction methods, preferably prepared by using solvent extraction or reflux extraction. can do. The extraction process may be repeated several times, and then additional steps such as concentration or lyophilization may be performed. Specifically, the obtained extract may be concentrated under reduced pressure to obtain a concentrate, and the concentrate may be lyophilized and then a high-concentration extract powder may be prepared using a grinder. The extract also includes fractions obtained by further fractionating the extract.

The extract may use at least one selected from the group consisting of water, organic solvents, supercritical fluids, and mixtures thereof as an extraction solvent. The organic solvent is an alcohol, preferably a C1-C4 lower alcohol, hexane (n-hexane), ether, glycerol, propylene glycol, butylene glycol, ethyl acetate, methyl acetate, dichloromethane, chloroform, ethyl acetate, benzene and It may be any one selected from the group consisting of these mixed solvents, preferably ethanol. For example, when a mixture of water and an organic solvent is used as the extraction solvent, the mixture of water and the organic solvent may preferably be a mixture of water and a C1-C4 lower alcohol, more preferably a mixture of water and ethanol. may be a mixture.

The mixture of water and ethanol may be a 3% (v/v) to 70% (v/v) ethanol aqueous solution, such as a 5% (v/v) to 50% (v/v) ethanol aqueous solution, preferably It may be a 10% (v/v) to 45% (v/v) ethanol aqueous solution, more preferably a 15% (v/v) to 40% (v/v) ethanol aqueous solution, and even more preferably may be a 20% (v/v) to 35% (v/v) ethanol aqueous solution, but is not limited thereto. When the concentration of the aqueous ethanol solution is less than the lower limit during preparation of Eucommia or dermal extract, it is difficult to extract the active ingredient exhibiting the preventive, ameliorative, or therapeutic effect of gastrointestinal disorders, making it difficult to obtain the desired degree of prevention, improvement, or treatment of gastrointestinal disorders.

Extraction may be carried out at 60 °C to 100 °C, preferably at 65 °C to 95 °C, more preferably at 70 °C to 90 °C, but is not limited thereto. Extraction may be carried out for 4 hours to 24 hours, preferably for 4 hours to 16 hours, more preferably for 4 hours to 12 hours, but is not limited thereto.

Extraction may be performed 1 to 7 times, preferably 1 to 5 times, and more preferably 1 to 3 times, but is not limited thereto, and the extract is obtained from each extraction. It may be a single extract or a mixed extract of extracts obtained from each extraction.

In order to obtain a specific active ingredient highly effective in preventing, improving or treating gastrointestinal disorders from each of the above extracts, fractions obtained by additionally performing a conventional fractionation process may be used. For example, the fraction obtained by passing the extract obtained according to the extraction through an ultrafiltration membrane having a certain molecular weight cut-off value, various chromatography (made for separation according to size, charge, hydrophobicity or affinity) Active fractions obtained through various additional purification methods such as separation are also included in the extract of the present invention. A specific active fraction having better preventive, ameliorative or therapeutic effects on gastrointestinal disorders can be prepared by separating an extract in which various components are mixed according to the nature of the active component through concentration gradient column chromatography or the like.

The chromatography may be, for example, at least one selected from the group consisting of column chromatography, high performance liquid chromatography (HPLC), ion exchange chromatography, affinity chromatography, and size exclusion chromatography.

The column chromatography is performed by column chromatography using a filler selected from the group consisting of silica gel, Sephadex, LH-20, ODS gel, RP-18, polyamide, Toyopearl and XAD resin to separate active fractions And it can be purified, column chromatography can be carried out several times by selecting an appropriate filler as necessary, but is not limited thereto. In using the chromatography, the elution solvent, elution rate, and elution time may apply solvents, rates, or times generally used in the art.

In addition, in order to obtain a specific active ingredient highly effective in preventing, improving or treating gastrointestinal disorders, fractions obtained by fractionating each of the above extracts with an organic solvent may be used. The organic solvent is alcohol, hexane (n-hexane), ether, glycerol, propylene glycol, butylene glycol, ethyl acetate, methyl acetate, dichloromethane, chloroform, ethyl acetate, benzene, acetone, acetonitrile and mixed solvents thereof It may be any one selected from the group consisting of, but is not limited thereto.

2. Pharmaceutical composition for prevention or treatment of gastrointestinal disorder comprising Eucommia extract and dermis extract

One aspect of the present invention provides a pharmaceutical composition for preventing or treating gastrointestinal disorders comprising an extract of Eucommia and a dermis extract.

The method for preparing the extract and dermis extract is the same as that described in " 1. Preparation of extract and dermis extract ", so the description is omitted.

The gastrointestinal disorder may be caused by excessive secretion of gastric acid, and may be used to include heartburn, gastritis, peptic ulcer such as gastric ulcer or duodenal ulcer, and reflux esophagitis. In addition, symptoms of excessive gastric acid secretion may occur due to mental anxiety, stress, irregular life, or chronic gastrointestinal disease, but are not particularly limited thereto.

In the present invention, "prevention" refers to all activities that suppress or delay gastrointestinal disorders by administration of the composition, and "treatment" refers to all activities that improve or beneficially change the symptoms of gastrointestinal disorders by the composition.

The pharmaceutical composition for preventing or treating gastrointestinal disorders of the present invention contains the Eucommia extract and the dermal extract in a weight ratio of 5:1 to 1:1, or a weight ratio of 4:1 to 2:1, or 3.5:1 to 2.5:1 It can be included in a weight ratio of If the ratio of the Eucommia extract and the dermis extract is out of the above range, the effect of preventing or treating gastrointestinal disorders may be lowered.

In addition, the mixture of the Eucommia extract and the dermal extract is 0.005 to 20.0% by weight, or 0.01 to 10.0% by weight, or 0.01 to 5% by weight, or 0.01 to 5% by weight, based on the total weight of the pharmaceutical composition for preventing or treating gastrointestinal disorders. It may be contained in an amount of 3% by weight, or 0.1 to 2% by weight, or 0.5 to 1.5% by weight, but is not limited thereto. If the effective content of the mixture of the Eucommia extract and the dermis extract is less than 0.005% by weight, the effect of preventing or treating gastrointestinal disorders is lowered, and if it exceeds 20% by weight, the effect of increasing the efficacy due to the increase in the content is reduced, which is uneconomical. can

The pharmaceutical composition of the present invention, in addition to the mixture of the Eucommia extract and the dermis extract, can give a synergistic effect to the effect of the mixture of the Eucommia extract and the dermis extract, preferably within a range that does not impair the desired effect of the present invention. It may additionally contain other ingredients that are present. For example, antioxidants, stabilizers, solubilizers, hydrating or emulsifying accelerators, vitamins, salts and/or buffers for osmotic pressure control, adjuvants such as, and other therapeutically useful substances may be included.

The pharmaceutical composition may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, etc., external preparations, suppositories and sterile injection solutions, respectively, according to conventional methods.

Solid preparations for oral administration include powders, granules, tablets, capsules, soft capsules, pills and the like. Liquid preparations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included. have.

Formulations for parenteral administration include powders, granules, tablets, capsules, sterilized aqueous solutions, solutions, non-aqueous solutions, suspensions, emulsions, syrups, suppositories, aerosols, etc., and sterile injection preparations according to conventional methods, respectively. It can be formulated and used in the form of a cream, gel, patch, spray, ointment, warning, lotion, liniment, pasta, or cataplasma. , but is not limited thereto. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents. As a base for the suppository, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogeratin and the like may be used.

The route of administration of the pharmaceutical composition of the present invention includes oral, intravenous, intramuscular, intraarterial, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal, for example topical administration by application (topical application) can be applied. The parenteral includes subcutaneous, intradermal, intravenous, intramuscular, intralesional injection or infusion techniques.

The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount.

In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is the type and severity of the subject, age, sex, type of disease, It may be determined according to the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the rate of excretion, the duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, or may be administered sequentially or simultaneously with conventional therapeutic agents. And it can be single or multiple administration. It is important to administer the amount that can obtain the maximum effect with the minimum amount without side effects in consideration of all of the above factors, and can be easily determined by a person skilled in the art.

The dosage of the pharmaceutical composition of the present invention may vary depending on various factors including age, body weight, general health, sex, administration time, route of administration, excretion rate, drug combination, and severity of a specific disease.

In addition, the pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers. In addition, the pharmaceutical composition of the present invention may be administered at a dosage within the range of 0.001 to 200 mg/kg on an adult basis, 70 to 200 mg/kg, or 80 to 170 mg/kg, or 130 to 160 mg/kg, or at a dose of 145 to 155 mg/kg. When the pharmaceutical composition is for external use, it is recommended to apply the pharmaceutical composition 1 to 5 times a day in an amount of 1.0 to 3.0 ml for adults and continue for at least one month, but the dosage is not intended to limit the scope of the present invention.

In addition, the pharmaceutical composition of the present invention is not particularly limited as long as it is a subject for the purpose of preventing or treating gastrointestinal disorders, and any may be applied. For example, the pharmaceutical composition of the present invention can be used not only for humans but also for non-human animals such as monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cows, sheep, pigs, goats, birds and fish. do.

3. Health functional food composition for preventing or improving gastrointestinal disorders containing Eucommia extract and dermis extract

One aspect of the present invention provides a health functional food composition for preventing or improving gastrointestinal disorders including Eucommia extract and dermal extract.

The method for preparing the extract and dermis extract is the same as that described in " 1. Preparation of extract and dermis extract ", so the description is omitted.

The gastrointestinal disorder may be caused by excessive secretion of gastric acid, and may be used to include heartburn, gastritis, peptic ulcer such as gastric ulcer or duodenal ulcer, and reflux esophagitis. In addition, symptoms of excessive gastric acid secretion may occur due to mental anxiety, stress, irregular life, or chronic gastrointestinal disease, but are not particularly limited thereto.

In the present invention, prevention or improvement of gastrointestinal disorder can be achieved by suppressing hypersecretion of gastric acid.

The health functional food composition for preventing or improving gastrointestinal disorders of the present invention contains the Eucommia extract and the dermal extract at a weight ratio of 5:1 to 1:1, or a weight ratio of 4:1 to 2:1, or 3.5:1 to 2.5: 1 in a weight ratio. If the ratio of the Eucommia extract and the dermis extract is out of the above range, the effect of preventing or improving gastrointestinal disorders may be lowered.

In addition, the mixture of the Eucommia extract and the dermal extract is 0.005 to 20.0% by weight, or 0.01 to 10.0% by weight, or 0.01 to 5% by weight, or 0.01% by weight based on the total weight of the health functional food composition for preventing or improving the gastrointestinal disorder. to 3% by weight, or 0.1 to 2% by weight, or 0.5 to 1.5% by weight, but is not limited thereto. If the effective content of the mixture of the Eucommia extract and the dermis extract is less than 0.005% by weight, the effect of preventing or improving gastrointestinal disorders is lowered, and if it exceeds 20% by weight, the effect of increasing the efficacy due to the increase in content is reduced, which is uneconomical. can

The health functional food composition of the present invention, in addition to the mixture of the Eucommia extract and the dermis extract of the present invention, has a synergistic effect on the effect of the mixture of the Eucommia extract and the dermis extract, preferably within a range that does not impair the effect intended by the present invention. It may additionally contain other ingredients that can give Conventional adjuvants such as, for example, stabilizers, solubilizers, vitamins, pigments and flavors, or carriers may be included.

In addition, it may include ingredients that are commonly added during the preparation of health functional food compositions, and include, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. Examples of such carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides such as conventional sugars such as dextrins and cyclodextrins and sugar alcohols such as xylitol, sorbitol and erythritol. As the flavoring agent, natural flavoring agent thaumatin, stevia extract, and synthetic flavoring agent may be used. For example, when the health functional food composition of the present invention is prepared as a drink, citric acid, high fructose corn syrup, sugar, glucose, acetic acid, malic acid, fruit juice, natural extract, etc. may be further included in addition to the Eucommia extract and the dermis extract of the present invention.

The health functional food to which the mixture of the extract of the present invention and the dermis extract can be added is not particularly limited, but, for example, meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gum, Dairy products including ice creams, various soups, beverages, tea, drinks, alcoholic beverages and vitamin complexes, etc., include all foods in a conventional sense.

The health functional food may be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. for the purpose of preventing or improving gastrointestinal disorders.

For example, a health functional food in the form of a tablet is obtained by granulating a mixture obtained by mixing the health functional food composition with excipients, binders, disintegrants, and other additives in a conventional manner, and then compression molding by adding a lubricant or the like. The mixture can be directly compression molded. In addition, the health functional food in the form of a tablet may contain a flavoring agent and the like as needed.

Among the health functional foods in the form of capsules, hard capsules can be prepared by filling a mixture obtained by mixing the health functional food composition with additives such as excipients in a normal hard capsule, and soft capsules can be prepared by filling the health functional food composition with additives such as excipients. It can be prepared by filling the mixture mixed with gelatin in a capsule base. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary.

The health functional food in the form of a pill can be prepared by molding a mixture obtained by mixing the health functional food composition with an excipient, a binder, a disintegrant, etc. by a conventionally known method, and can be coated with sucrose or other coating agent if necessary. , or the surface may be coated with a material such as starch or talc.

Health functional food in the form of granules can be prepared by a previously known method of a mixture of the health functional food composition and excipients, binders, disintegrants, etc. in granular form, and, if necessary, may contain flavoring agents, flavoring agents, etc. can

Hereinafter, the present invention will be described in detail with reference to preparation examples and examples.

However, the following Preparation Examples, Experimental Examples, and Examples are only for exemplifying the present invention, and the contents of the present invention are not limited by the following Preparation Examples and Examples.

Preparation Example 1. Preparation of Eucommia extract

Eucommia ulmoides Oliver ( Eucommia ulmoides Oliver , stem bark, 2 kg) was added with a 10-fold 30% (v / v) ethanol aqueous solution compared to the weight, and extracted twice at 80 ° C. for 4 hours. An extract obtained by mixing the extracts obtained in each extraction was filtered using a 5 μm filter. The filtrate was concentrated at 55° C. using a rotary vacuum concentrator (BUCHI, R-220). Finally, the concentrate was diluted to 15 °Brix by adding sterilized distilled water, and then lyophilized (OPERON, FDT-12012) to obtain 255.9 g of Eucommia extract powder.

Preparation Example 2. Preparation of dermal extract

A 30% (v/v) aqueous solution of 18 times the weight of ethanol was added to 1 kg of the dermis prepared by drying the rind of the fruit of the tangerine tree ( Citrus unshiu Markovich), extracted twice for 8 hours at 80 ° C, and the extract was 5㎛ It was filtered using a filter. The filtrate is concentrated at 50~60℃ using a rotary vacuum condenser (BUCHI, R-220), and finally diluted to 15°Brix by adding sterilized distilled water to the concentrate, and freeze-dried (OPERON, FDT-12012). 447 g of dermal extract powder was obtained.

Preparation Example 3. Preparation of a composite of Eucommia extract and dermal extract

Three types of composites were prepared by mixing the single extracts obtained in Preparation Example 1 and Preparation Example 2 at different ratios. In each composite, the weight ratio of Eucommia:dermis was 3:1, 1:1, and 1:3.

Example 1. Efficacy test for improving hypersecretion of gastric acid in histamine-induced gastric parietal cells

1-1. Gastric acid secreting factor cAMP level measurement

After treating gastric parietal cells with the single extracts and complexes of Preparation Examples 1 to 3 at a concentration of 100 μg/ml, histamine (Sigma, H7125, USA) was treated with gastric parietal cells at a concentration of 100 μM and cultured for 24 hours, followed by cAMP ELISA kit ( Cell biolabs) was used according to the standard reagents and analysis procedures provided by the manufacturer. After removing the media by selecting the non-acetylated version (Version), 1 ml of lysis buffer is dispensed per 35 cm ² . At this time, the lysis buffer is dispensed in proportion to each area. After dispensing the lysis buffer, the mixture was reacted at 4° C. for 20 minutes, scraped the surface of the plate with a cell scraper, homogenized using a pipette, and centrifuged for 10 minutes, and the supernatant was used as a sample for the experiment. 50 μl of the supernatant and 50 μl of cAMP standard reagent are dispensed for each concentration on the goat anti-rabbit antibody-coated plate. Thereafter, 25 μl of the diluted peroxidase cAMP tracer conjugate was dispensed, covered with a plate cover, and reacted while shaking in an orbital shaker at room temperature for 2 hours. After 2 hours, 250 μl of 1X washing buffer was dispensed into each well to wash 5 times, and 100 μl of substrate solution was dispensed and reacted at room temperature for 5 to 20 minutes while shaking using an orbital shaker. Thereafter, the stop solution was dispensed by 50 μl, and the OD value was measured using an ELISA reader, and the amount was calculated using a standard curve, and is shown in Table 1 below.

cAMP level (pmol/ml) Normal group (NC) 0.49 control group (C) 2.36 Eucommia 2.01 Doujin (3:1) 1.49 Doujin (1:1) 1.93 Doujin (1:3) 1.88 dermis 1.69

In the results of Table 1, the cAMP level of the control group not treated with the sample was set as 100%, and the result of confirming the cAMP level (%) of the experimental group is shown in FIG. 1 .

From the above results, it can be confirmed that the cAMP level was significantly improved in the group treated with 100 μg/ml of the compound obtained by mixing the Eucommia extract and the dermal extract at a ratio of 3:1.

1-2. Gastric acid secreting factor H+/K+ ATPase activity measurement

After treating gastric parietal cells with the single extracts and complexes of Preparation Examples 1 to 3 at a concentration of 100 μg/ml, histamine (Sigma, H7125, USA) was treated with gastric parietal cells at a concentration of 100 μM and cultured for 24 hours, followed by H + / K + ATPase It was performed according to standard reagents and analysis procedures provided by the manufacturer using an analysis kit (Elabscience).

Reagents 1 to 11 configured in the kit are prepared to be used in the experiment after pretreatment according to the protocol, and each reagent is added to the sample control tube to exclude the experimental value by identifying the unique color development level of the sample tube and sample. . Dispense 130 μl of reagent 1 into the sample control tube and sample tube, 80 μl of reagent 2 into the sample tube, 120 μl of reagent 3 into the sample control tube, and 120 μl of reagent 4 into the sample control tube and sample tube. 40 μl each, 40 μl each of reagent No. 5 into the sample control tube and sample tube, 40 μl of reagent No. 6 into the sample tube, and 100 μl of the sample to be tested into the sample tube and mixing. 37 React in a water bath at °C for 10 minutes. Thereafter, 50 μl of reagent No. 7 was dispensed into each of the sample control tube and the sample tube, and 100 μl of the sample to be tested was dispensed into the sample control tube, centrifuged at 3500 rpm for 10 minutes, and the supernatant was used for phosphorus analysis. use. Dispense 400 μl of the 0.5 μmol/ml standard phosphorus solution into the standard tube, dispense 400 μl of the supernatant from the sample control tube reacted into a new sample control tube, and dispense 400 μl of the supernatant from the sample tube reacted into a new sample tube. Dispense 2000 μl each of the phosphorus analysis reagent into a new sample control tube and sample tube, react in a water bath at 45 ° C for 5 minutes, measure the O.D value of 660 nm using an elisa reader, and subtract the sample O.D value from the sample control O.D. Values were calculated and shown in Table 2 below.

H+/K+ ATPase activity
(U/mg prot) Normal group (NC) 1.7 control group (C) 7.4 Eucommia 5.6 Doujin (3:1) 5.0 Doujin (1:1) 3.7 Doujin (1:3) 4.4 dermis 4.8

In the results of Table 2, the H + / K + ATPase activity of the control group not treated with the sample was set as 100%, and the results of confirming the H + / K + ATPase activity (%) of the experimental group are shown in FIG. 2 .

From the above results, it can be confirmed that the H+/K+ ATPase activity was significantly improved in the group treated with 100 μg/ml of the compound obtained by mixing the Eucommia extract and the dermal extract at a ratio of 3:1.

Example 2. Efficacy test for improving gastric acid secretion in alcoholic gastric injury animal models

2-1. Composition of experimental group

Nam et al., J. Korean Soc. Food Sci. Nutr. , 43 (10), 150-1509 (2014), the experiment was performed by applying the method disclosed in the literature. The normal and experimental groups were supplied with 7-week-old male SD rats from Saeron Bio Co., Ltd. (Gyeonggi-do, Korea) and acclimatized for 7 days. (55±10%) and light/dark cycle (12 hours) were automatically adjusted. After stabilization, the weight was measured to separate the groups, and the experiment was conducted (Table 3).

number label number of horses sample dose stomach-damaging substances One Normal group (NC) 6 - - 2 control group (C) 6 - 40% EtOH / 100 mM HCl 1 ml each 3 Eucommia 6 Eucommia 100 mg/kg 4 dermis 6 Dermis 100 mg/kg 5 Doujin (3:1) 6 Eucommia + dermis (3:1) 100 mg/kg 6 Doujin (1:1) 6 Eucommia + dermis (1:1) 100 mg/kg 7 Doujin (1:3) 6 Eucommia + dermis (1:3) 100 mg/kg

After group separation, once a day for 2 weeks, single extracts and complexes of Preparation Examples 1 to 3 and 40% ethanol were orally administered to each animal in an amount of kg / bw as shown in Table 3 using an oral sonde. After oral administration for a total of 2 weeks was completed, 100 mM HCl was administered after 24 hours of fasting, and 1 hour later, autopsy was performed, blood was collected from the hepatic vein, and gastric tissue was removed.

2-2. Gastric juice secretion measurement

Before taking a picture of the extracted stomach, a small incision was made in the pylorus, gastric juice was collected in a 50 ml conical tube, centrifuged at 3,000 rpm for 20 minutes, and the supernatant was collected to measure the amount of gastric juice secretion (ml), and the results are shown in Table 4. showed up

Gastric juice secretion (ml) Normal group (NC) 0.44 control group (C) 3.05 Eucommia 2.41 Doujin (3:1) 1.72 Doujin (1:1) 2.10 Doujin (1:3) 1.97 dermis 2.67

In addition, in the results of Table 4, the gastric juice secretion amount of the control group not treated with the sample was set as 100%, and the gastric juice secretion amount (%) of the experimental group was confirmed. The results are shown in FIG. 3 .

From the above results, it can be confirmed that the gastric juice secretion was significantly improved in the group administered with 100 mg/kg of the compound obtained by mixing the Eucommia extract and the dermal extract at a ratio of 3:1.

2-3. Gastric wall damage index measurement

The extracted stomach was incised along the large intestine, washed the surface of the gastric wall with saline, spread well on a 6-well plate using a cotton swab or tweezers, fixed with formalin, and photographed. For the photograph taken, the gastric damage area and total stomach area were measured using an image program (Image J, USA), and the gastric wall damage index (Ulcer Index) (%) was obtained according to the following formula, and the results are shown in Table 5. .

Gastric wall damage index (%) = damaged area/total area.

Gastric wall damage index (%) Normal group (NC) 0.67 control group (C) 6.62 Eucommia 5.05 Doujin (3:1) 3.50 Doujin (1:1) 5.39 Doujin (1:3) 4.22 dermis 6.41

In addition, in the results of Table 5, the gastric wall damage index of the control group not treated with the sample was set as 100%, and the results of confirming the gastric wall damage index (%) of the experimental group are shown in FIG.

From the above results, it can be confirmed that the group administered with 100 mg/kg of a compound obtained by mixing Eucommia extract and dermal extract at a ratio of 3:1 significantly improved gastric wall damage.

2-4. Measurement of histamine levels in the blood

Blood histamine level was measured using a histamine ELISA kit (abcam) according to standard reagents and analysis procedures provided by the manufacturer. After preparing 500 μl of serum, it was dried using a centrifugal concentrator, treated with 500 μl of assay buffer, and used as a sample for the experiment. After dispensing 100 μl of the sample into the microplate, 50 μl of 1x histamine tracer is dispensed. Thereafter, 50 μl of 1x histamine antibody was dispensed and reacted while mixing for 1 hour using an orbital shaker. After washing three times using the washing buffer, 200 μl of SA-HRP was dispensed and reacted while mixing for 30 minutes using an orbital shaker. After washing three times using the washing buffer, 200 μl of the TMB substrate solution was dispensed and reacted while mixing for 30 minutes using an orbital shaker. After 30 minutes, 50 μl of the stop solution was dispensed, and the O.D value was measured and calculated at a wavelength of 450 nm using an ELISA reader, and the results are shown in Table 6.

Blood histamine content (μg/ml) Normal group (NC) 1.15 control group (C) 3.49 Eucommia 2.74 Doujin (3:1) 2.53 Doujin (1:1) 3.38 Doujin (1:3) 2.70 dermis 2.98

In addition, in the results of Table 6, the blood histamine level of the control group not treated with the sample was set as 100%, and the result of confirming the blood histamine level (%) of the experimental group is shown in FIG. 5 .

From the above results, it can be confirmed that the group administered with 100 mg/kg of the compound obtained by mixing the Eucommia extract and the dermal extract at a ratio of 3:1 significantly improved the histamine content in the blood.

Preparation Example 4. Preparation of pharmaceutical composition

4-1. manufacture of powders

2 g of a mixture of Eucommia extract and dermal extract of the present invention

1 g lactose

A powder was prepared by mixing the above components and filling in airtight bags.

4-2. manufacture of tablets

100 mg of a mixture of Eucommia extract and dermis extract of the present invention

Corn Starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above ingredients, tablets were prepared by tableting according to a conventional tablet manufacturing method.

4-3. Manufacture of capsules

100 mg of a mixture of Eucommia extract and dermis extract of the present invention

Corn Starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above ingredients, capsules were prepared by filling gelatin capsules according to a conventional capsule preparation method.

4-4. manufacture of rings

1 g of a mixture of Eucommia extract and dermal extract of the present invention

lactose 1.5 g

1 g of glycerin

0.5 g xylitol

After mixing the above components, it was prepared to be 4 g per pill according to a conventional method.

4-5. Preparation of granules

150 mg of a mixture of Eucommia extract and dermis extract of the present invention

Soybean Extract 50 mg

Glucose 200 mg

Starch 600 mg

After mixing the above components, 100 mg of 30% ethanol was added and dried at 60° C. to form granules, which were then filled into bags.

Preparation Example 5. Manufacture of health functional food

5-1. Manufacture of Flour Food

0.5 to 5.0 parts by weight of a mixture of the extract of the present invention and the extract of the dermis was added to wheat flour, and bread, cakes, cookies, crackers and noodles were prepared using the mixture.

5-2. Preparation of soups and gravies

0.1 to 5.0 parts by weight of the mixture of the extract of the present invention and the extract of the dermis was added to soup and broth to prepare meat products and noodle soup and broth.

5-3. Manufacture of ground beef

Ground beef was prepared by adding 10 parts by weight of a mixture of the Eucommia extract and the dermis extract of the present invention to ground beef.

5-4. Manufacture of dairy products

5 to 10 parts by weight of the mixture of the extract of the present invention and the dermis extract was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

5-5. manufacture of wire

Brown rice, barley, glutinous rice, and adlay were alphanized by a known method, dried, roasted, and then prepared into a powder having a particle size of 60 mesh using a grinder.

Black beans, black sesame seeds, and perilla seeds were also steamed and dried by a known method, roasted, and then prepared into powder with a particle size of 60 mesh by a grinder.

A mixture of the extract of the present invention and the extract of the dermis was concentrated under reduced pressure in a vacuum concentrator, dried with a spray or hot air dryer, and the dried product was pulverized with a grinder to a particle size of 60 mesh to obtain a dry powder.

It was prepared by blending the mixture of the grains, seeds, and Eucommia extract of the present invention and the dermis extract prepared above in the following ratio.

Cereals (30 parts by weight of brown rice, 15 parts by weight of adlay, 20 parts by weight of barley), seeds (7 parts by weight of perilla seeds, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds), a mixture of Eucommia extract and dermal extract of the present invention (3 parts by weight) parts by weight), Ganoderma lucidum (0.5 parts by weight), Rehmannia Root (0.5 parts by weight)

5-6. Manufacture of health functional beverages

Homogeneously blending 5 g of a mixture of the extract of the present invention and the dermis extract with auxiliary ingredients such as high-fructose corn syrup (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%), and water (75%) After instantaneous sterilization, it was prepared by packaging in a small container such as a glass bottle or a plastic bottle.

5-7. Preparation of vegetable juice

Vegetable juice was prepared by adding 5 g of a mixture of the Eucommia extract and the dermis extract of the present invention to 1,000 ml of tomato or carrot juice.

5-8. manufacture of fruit juice

Fruit juice was prepared by adding 1 g of a mixture of the Eucommia extract and the dermis extract of the present invention to 1,000 ml of apple or grape juice.

Claims (10)

A pharmaceutical composition for preventing or treating gastrointestinal disorders comprising an extract of Eucommia and a dermis extract. The method of claim 1,
A composition comprising the Eucommia extract and the dermal extract in a weight ratio of 4:1 to 2:1. The method of claim 1,
A composition comprising the Eucommia extract and the dermal extract in a weight ratio of 3.5: 1 to 2.5: 1. The method of claim 1,
When preparing the Eucommia extract, the extraction solvent is water or a C1-C4 alcohol composition. The method of claim 1,
When preparing the dermal extract, the extraction solvent is water or a C1-C4 alcohol composition. The method of claim 1,
The composition for preventing or treating the gastrointestinal disorder is achieved by inhibiting hypersecretion of gastric acid. The method of claim 1,
The gastrointestinal disorder composition comprising at least one selected from the group consisting of heartburn, gastritis, gastric ulcer, duodenal ulcer and reflux esophagitis. A health functional food composition for preventing or improving gastrointestinal disorders comprising an extract of Eucommia and a dermis extract. The method of claim 8,
A composition comprising the Eucommia extract and the dermal extract in a weight ratio of 4:1 to 2:1. The method of claim 8,
The gastrointestinal disorder composition comprising at least one selected from the group consisting of heartburn, gastritis, gastric ulcer, duodenal ulcer and reflux esophagitis.

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