WO2014042426A1 - Composition comprising saikosaponin a, berberine, and licoisoflavone b for preventing or treating gastric diseases - Google Patents

Abstract

The present invention relates to a composition comprising saikosaponin A, berberine, and licoisoflavone B for preventing or treating gastric diseases, and more particularly, to a pharmaceutical composition for preventing or treating gastric diseases, to a food composition for preventing or treating gastric diseases, to a method for preparing the composition, and to a method for treating gastric diseases using the composition, wherein the pharmaceutical composition and the food composition comprise 15 mg/g to 50 mg/g of saikosaponin A, 30 mg/g to 100 mg/g of berberine, and 0.5 mg/g to 5 mg/g of licoisoflavone B.

Description

Composition for the prophylaxis or treatment of gastrointestinal diseases, including psychosaponin A, berberine and lysoisoflavone Ｂ

The present invention relates to a composition for the prevention or treatment of gastrointestinal diseases, including psychosaponin A (Saikosaponin A), berberine (Berberine) and lycoisoflavone B (Licoisoflavone B), specifically 15 mg / g to 50 mg / g, berberine 30mg / g to 100mg / g and lysoisoflavone B 0.5mg / g to 5mg / g, a pharmaceutical composition for the prevention or treatment of gastrointestinal diseases, food compositions for the prevention or improvement of gastrointestinal diseases, the composition It relates to a method for preparing and a method for treating gastrointestinal diseases using the composition.

The causes of gastritis and gastric ulcer are largely caused by pathological excess of attack factors such as gastric acid, pepsin, alcohol and smoking, stress and nonsteroidal anti-inflammatory drugs, Helicobacter pylori infection, and gastric mucosal cells from gastric acid attack. It may be classified into a decrease in the secretion of bicarbonate ions, a protective protective factor, a decrease in the production of prostaglandins, and a structural or morphological defect of the gastric mucosa. In order to treat inflammation and ulcer of stomach and duodenum caused by attack factors, gastric acid secretion, mucous secretion promotion, gastric mucosal epithelial cell regeneration, proliferation of Helicobacter pylori bacteria and anti-inflammatory drugs are required.

H2-receptor antagonist (H2-RA), the most representative treatment for gastritis and gastric ulcer, is cimetidine, ranitidine, famotidine, roxatidine, Although nizatidine has excellent anti-ulcer effect in clinical practice, relapse occurs frequently after discontinuation of drug administration, and in case of long-term administration, the drug continues to accumulate in the body and stop the medication. Or side effects such as thickening of the stomach wall by dosing have been reported. Therefore, the use of only H2 inhibitors and proton-pump inhibitors to suppress gastric acid secretion for the treatment of gastritis and gastric ulcers cannot suppress the recurrence of lesions that are most problematic in the treatment of peptic ulcers. If it progresses, it will not be possible to take it for a long time. In addition, cimetidine is known to exhibit side effects such as neutropenia and drug interactions. Recently, the use of cimetidine has been shown to avoid its use. Recently, drugs such as ranitidine and famotidine have a significant decrease in efficacy. Was reported.

In addition, proton pump inhibitors (PPIs) include omeprazole, omeprazole, lansoprazol, and pantoprazole, which inhibit acid secretion in gastric parietal cells in the final stages, and have a strong effect of inhibiting acid secretion. It is known to represent.

Other therapeutic agents that can be used to treat gastritis and gastric ulcers include prostaglandin biosynthesis-promoting gastric mucosal diseases such as antacid, Artemisiae argyi Folium extract, and teprenone. There are drugs such as rebamipide. Gastric mucosal protectors are known to regenerate mucosal tissue to a similar level as normal to compensate for the short-acting gastric acid secretion inhibitors (Folia Pharmacol. Japan, 92: 389, 1988) and are an important part of the treatment of gastritis. . However, gastric mucosa protective agents have the disadvantage that they are fast-acting, high-dose, and have to be taken for a relatively long time.

Currently, various types of peptic ulcer drugs have been developed, but these drugs only play a role in suppressing the attack from gastric acid and thus keeping the wound from worsening, and they are essentially a direct healing effect to heal wounds of peptic ulcers. It does not show, the disappearance of the ulcers will eventually have to rely on the natural healing power of the human body. Therefore, various studies have been conducted to develop a therapeutic agent using herbal extracts that can shorten the healing period and reduce the recurrence rate in consideration of the limitations of the existing peptic ulcer therapeutic agent, and a composition for treating gastrointestinal diseases including mugwort leaf extract as an active ingredient. (Korean Patent No. 10-0181751) and the like have been commercialized as styrene tablets and sold and prescribed as a successful natural gastritis treatment.

Accordingly, the present inventors have investigated the active ingredient from the mixed herbal extracts having a gastrointestinal disease treatment effect, and have tried hard to determine the optimal composition ratio with excellent gastrointestinal disease effects but no side effects, and the psychosaponin A, berberine and lysoisoflavones. It was confirmed that the three B components significantly bring about the therapeutic effect of gastrointestinal diseases, and the optimal composition ratio was identified to complete the present invention.

One object of the present invention is 15 mg / g to 50 mg / g of Saikosaponin A, 30 mg / g to 100 mg / g of Berberine and 0.5 mg / g to 5 mg / g of Lycoisoflavone B. It includes, to provide a pharmaceutical composition for the prevention or treatment of gastrointestinal diseases.

Another object of the present invention is a food for preventing or ameliorating gastrointestinal diseases, including 15 mg / g to 50 mg / g of psychosaponin A, berberine 30 mg / g to 100 mg / g, and 0.5 mg / g to 5 mg / g of lysoisoflavone B. It is to provide a composition.

Another object of the present invention is to provide a method for preparing the composition.

Another object of the present invention to provide a method for treating gastrointestinal diseases using the composition.

Another object of the present invention is the prevention of gastrointestinal diseases of psychosaponin A 15mg / g to 50mg / g, berberine 30mg / g to 100mg / g and lysoisoflavone B 0.5mg / g to 5mg / g, or a composition comprising the same Or to provide a therapeutic use.

Still another object of the present invention is to provide a gastrointestinal disease of 15 mg / g to 50 mg / g, berberine 30 mg / g to 100 mg / g and lysoisoflavone B 0.5 mg / g to 5 mg / g, or a composition comprising the same. To provide a pharmaceutical manufacturing use for the prevention or treatment.

The composition of the present invention comprising psychosaponin A, berberine and lysoisoflavone B is composed of an optimal composition ratio with excellent therapeutic effect and fewer side effects, and can be usefully used for the prevention or treatment of gastrointestinal diseases.

1 is a view showing a process for producing a solvent-specific fraction of the mixed herbal extract of the present invention.

Figure 2 shows the schematic diagram of the drug test method for the animal model that caused gastrointestinal diseases with alcohol.

Figure 3 is a diagram showing the result of confirming the inhibitory effect of the mixed herbal extract of the present invention and the solvent fractions thereof for the animal model causing the gastrointestinal disease with alcohol.

Figure 4 is a diagram showing the results of gastritis inhibition rate when the administration of psychosaponin A, berberine and lysoisoflavone B, which are the active ingredients of the mixed herbal extract of the present invention in an animal model causing gastrointestinal diseases with alcohol, respectively.

Figure 5a shows the chemical structure of psychosaponin A, an active ingredient present in the mixed herbal extract of the present invention.

Figure 5b shows the chemical structure of berberine which is an active ingredient present in the mixed herbal extract of the present invention.

Figure 5c shows the chemical structure of lycoisoflavone B as an active ingredient present in the mixed herbal extract of the present invention.

In one embodiment, the present invention provides 15 mg / g to 50 mg / g of Saikosaponin A, 30 mg / g to 100 mg / g of Berberine, and 0.5 mg / g to 5 mg / g of Lycoisoflavone B. It provides a pharmaceutical composition for preventing or treating gastrointestinal diseases, comprising g.

The composition for preventing or treating gastrointestinal diseases of the present invention comprises psychosaponin A, berberine and lysoisoflavone B as active ingredients, and each of the psychosaponin A, berberine and lysoisoflavone B 15 mg / g to 50 mg / g, If it contains 30mg / g to 100mg / g and 0.5mg / g to 5mg / g, it has an excellent effect in the prevention or treatment of gastrointestinal diseases. Said psychosaponin A, berberine, lycoisoflavone B can be obtained by extracting and separating from various plants including the compound by a commercialized compound or a known method.

In the present invention, the term "saikosaponin A" is a compound having the structure of Formula 1 below, (3beta, 4alpha, 16beta) -13,28-Epoxy-16,23-dihydroxyolean-11-en-3 It may also be named -yl-6-deoxy-3-O-beta-D-glucopyranosyl-beta-D-galactopyranoside.

Formula 1

For the purposes of the present invention, the psychosaponin A refers to a compound having an effect in the prevention or treatment of gastrointestinal diseases.

As used herein, the term "berberine" refers to a quaternary ammonium salt belonging to the protoberberine group of the isoquinoline alkaloid, and has a structure of Formula 2 below.

Formula 2

For the purpose of the present invention, the berberine may mean a compound which is included as an active ingredient in the composition of the present invention and may help in preventing or treating gastrointestinal diseases, but is not limited thereto.

In the present invention, the term "Licoisoflavone B (Licoisoflavone B)" refers to a compound having a structure of Formula 3 as a flavonoid-based compound.

Formula 3

Licoisoflavone B may also be named 5,7-dihydroxy-3- (5-hydroxy-2,2-dimethylchromen-6-yl) chromen-4-one.

The composition of the present invention may preferably comprise 15 mg / g to 50 mg / g, 30 mg / g to 100 mg / g and 0.5 mg / g to 5 mg / g, respectively, of the psychosaponin A, berberine and lycoisoflavone B . In one embodiment of the present invention, the present inventors have identified 12 active ingredients from the extract of the mixed herbal medicine having a therapeutic effect of gastrointestinal diseases, 3 of the present invention showing an excellent therapeutic effect on gastrointestinal diseases among the 12 active ingredients The compound of the species was identified (Experimental Example 1-3). In addition, it was confirmed that when the three compounds are included in the composition ratio, it can have an excellent therapeutic effect in gastrointestinal diseases.

In addition, the composition of the present invention comprising the psychosaponin A, berberine and lycoisoflavone B of the present invention may be in the form of a mixed herbal extract.

As used herein, the term "mixed herbal extract" may be in the form of an extract obtained by mixing natural products such as plants, followed by extraction with one or more solvents, or a mixture of extracts obtained by extracting each natural product with a solvent. This is not restrictive. The type of the solvent for extracting the extract from the mixed herbal extract is not particularly limited as long as it extracts the psychosaponin a, berberine and lysoisoflavone B, preferably water, alcohol having 1 to 6 carbon atoms and their Can be extracted with one or more solvents selected from the group consisting of mixed solvents, but is not limited thereto. In one embodiment of the present invention, 15 mg / g to 50 mg / g, 30 mg / g to 100 mg / g and 0.5 mg / g to 5 mg / g, respectively, of psychosaponin A, berberine and lysoisoflavone B in the form of a mixed herbal extract A composition was prepared.

The composition of the present invention comprising the psychosaponin A, berberine and lysoisoflavone B can be usefully used for the prevention or treatment of gastrointestinal diseases.

In the present invention, the term "gastrointestinal disease" is a generic term for diseases of the gastrointestinal tract, the gastrointestinal disease is not particularly limited if the gastrointestinal diseases that can be treated by the composition of the present invention, for example, gastritis, gastric ulcer, Duodenal ulcer, Solinger-Elison syndrome, reflux esophagitis, postoperative ulcer, erosion of gastric mucosa, bleeding of gastric mucosa, redness of gastric mucosa or edema of gastric mucosa, but is not limited thereto. In addition, the gastrointestinal disease may be characterized in that the mucous membrane damage of the stomach or duodenum caused by alcohol, smoking, stress, drugs, or a combination thereof, but is not limited thereto.

As used herein, the term "prevention" refers to any action that inhibits or delays the onset of gastrointestinal disease by administration of the composition, and "treatment" improves or advantageously changes the symptoms caused by gastrointestinal disease by administration of the composition. It means every act to do

The pharmaceutical composition comprising the psychosaponin A, berberine and lycoisoflavone B may further include appropriate carriers, excipients or diluents commonly used in the manufacture of pharmaceutical compositions, and may also include tablets, pills, powders, Granules, capsules, suspensions, solutions, emulsions, syrups, sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations and suppositories, and may have any one oral or parenteral formulation. There may be various formulations of the spheres. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which form at least one excipient such as starch, calcium carbonate, sucrose or lactose ( Lactose), gelatin, etc. are mixed. In addition to simple excipients, lubricants such as magnesium stearate, talc and the like are also used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, and syrups, and various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin, may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, etc. may be used. As the base of the suppository, Whitepsol, macrogol, Tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

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

As used herein, the term "administration" means introducing a pharmaceutical composition of the present invention to an individual in any suitable manner, and the route of administration of the composition is as long as possible to reach the desired tissue or through various routes of oral oral use. It may be administered, and specifically, in a conventional manner via the oral, rectal, topical, intravenous, intraperitoneal, intramuscular, intraarterial, transdermal, nasal, inhaled or intradermal routes.

As used herein, the term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level refers to the subject's type and severity, age, sex, and gastrointestinal disease. Can be determined according to the type of drug, activity of the drug, sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including concurrently used drugs, and other factors well known in the medical field. The compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents and may be administered sequentially or simultaneously with conventional therapeutic agents. And single or multiple administrations. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect in a minimum amount without side effects, and can be easily determined by those skilled in the art.

The composition can be administered to a variety of mammals, such as mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or Intracerebroventricular injection.

In one embodiment of the present invention, among the 12 active ingredients included in the extract of the mixed herbal medicine having a therapeutic effect of gastrointestinal diseases, three species of psychosaponin A, berberine and lysoisoflavone B show high gastric protective effect in gastritis animal model. It was confirmed (Table 3 and Figure 4). In particular, the extracts of Examples 1 to 4 comprising 15 mg / g to 50 mg / g of psychosaponin A, 30 mg / g to 100 mg / g of berberine, and 0.5 mg / g to 5 mg / g of lysoisoflavone B have excellent gastric protective effect. On the other hand (Tables 5 and 6, Fig. 3), Comparative Examples 1 to 3 containing 15 mg / g or less of psychosaponin and 30 mg / g or less of berberine had a drug efficacy of 23, 13 and -3%, respectively, about 40 to 56%. Compared with Examples 1 to 4 showing the efficacy of% showed a significantly lower efficacy (Table 7). Such results indicate that the composition of the present invention comprising psychosaponin A 15 mg / g to 50 mg / g, berberine 30 mg / g to 100 mg / g, and lysoisoflavone B 0.5 mg / g to 5 mg / g efficiently contains gastritis. It can be used to treat various gastrointestinal diseases. Therefore, the composition of the present invention can be effectively used for the prevention or treatment of various gastrointestinal diseases.

In another aspect, the present invention provides a method for treating gastrointestinal disease, comprising administering the pharmaceutical composition to a suspected gastrointestinal disorder.

The pharmaceutical composition and gastrointestinal diseases are as described above.

In particular, the method of treatment of the present invention comprises administering the pharmaceutical composition in a pharmaceutically effective amount into a suspected gastrointestinal disorder. The subject means an entire mammal including a dog, cow, horse, rabbit, mouse, rat, chicken or human, but the mammal of the present invention is not limited to the above examples. The pharmaceutical composition may be administered orally, orally, subcutaneously, intraperitoneally, pulmonary, and intranasally, and may be administered by a suitable method including topical administration if necessary for local treatment. Non-oral infusions include intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration. Preferred dosages of the pharmaceutical compositions of the present invention vary depending on the condition and weight of the individual, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art.

In another embodiment, the present invention provides for the prevention or amelioration of gastrointestinal diseases, including 15 mg / g to 50 mg / g of psychosaponin A, 30 mg / g to 100 mg / g of berberine and 0.5 mg / g to 5 mg / g of lysoisoflavone B. It provides a food composition for.

Said psychosaponin A, berberine, lycoisoflavone B and gastrointestinal diseases are as described above. More specifically, the psychosaponin A, berberine and lysoisoflavone B of the present invention comprises 15 mg / g to 50 mg / g, 30 mg / g to 100 mg / g and 0.5 mg / g to 5 mg / g, respectively. Or it may be added to the food composition for the purpose of improvement.

When the psychosaponin A, berberine and lysoisoflavone B are used as food additives, the compound may be added as it is or used with other foods or ingredients, and may be appropriately used according to a conventional method.

There is no particular limitation on the type of food of the present invention. Examples of foods to which the compounds may be added include meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, dairy products including other noodles, gums, ice creams, various soups, drinks, teas, drinks, Alcoholic beverages and vitamin complexes, and the like, and may include all foods in a conventional sense, and include foods used as feed for animals.

In addition to the above, the food composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols. And carbonation agents used in carbonated beverages. Others may contain pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks. In addition, the food may also be prepared in the form of tablets, granules, powders, capsules, liquid solutions and pills according to known production methods. There is no particular limitation on other components except for including the compound according to the present invention as an active ingredient, and various conventional flavoring agents or natural carbohydrates may be included as additional ingredients.

In addition to the above, the food composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols. And carbonation agents used in carbonated beverages. Others may contain pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components can be used independently or in combination.

In another aspect, the present invention provides a method of preparing the composition.

The method preferably comprises (a) 10 to 30 parts by weight of Coptidis Rhizoma and 10 to 30 parts by weight of licorice ( Glycyrrhizae Radix ) based on 100 parts by weight of Bupleuri Radix in water, 1 carbon. Extracting with at least one solvent selected from the group consisting of alcohols from 6 to 6 and mixed solvents thereof; (b) concentration of the filtrate obtained in the extraction step under reduced pressure; And (c) suspending by adding alcohol to the reduced pressure concentrate of step (b), and then preparing a soft-core extract by concentrating the secondary under reduced pressure, but is not limited thereto.

In the manufacturing method of the present invention, the solvent in the step (a) is preferably used 3 to 10 times the solvent compared to the mixed herbal weight, but is not limited thereto. In one embodiment of the present invention, about 5 times the weight of the mixed herbal weight ethanol was used as the solvent.

In addition, the method is preferably extracted for 20 to 100 hours at an extraction temperature of 40 to 50 ℃, and concentrated under reduced pressure at a temperature of 60 ℃ or less, but is not limited thereto. The vacuum concentration may be preferably performed at a temperature of 40 ° C. to 60 ° C., but is not limited thereto.

 In one embodiment of the present invention was extracted from the mixed herbal medicine for 22, 48, 91 hours at a temperature of 45 ℃, concentrated under reduced pressure to a concentration of 60 ℃ or less. Extracts prepared by the above method of a mixed herbal medicine containing 10 to 30 parts by weight of Cooptidis Rhizoma and 10 to 30 parts by weight of Glycyrrhizae Radix based on 100 parts by weight of Buleuri Radix were prepared by the following methods: psychosaponin A, berberine and lyco It was confirmed that isoflavone B contained 15 to 50 mg / g, 30 to 100 mg / g, and 0.5 mg / g to 5 mg / g, respectively, and it was confirmed that it had a therapeutic effect in gastrointestinal diseases (Tables 5 and 6).

In addition, the first reduced pressure concentration of step (b) of the method may be 10 to 20% by volume by drying under reduced pressure at a temperature of 500 to 700mmHg and 60 ℃ or less, but is not limited thereto. The vacuum drying may be preferably performed at a temperature of 40 ° C. to 60 ° C., but is not limited thereto. In one embodiment of the present invention was concentrated under reduced pressure at 500mmHg. The second vacuum concentration in step (c) may be dried under reduced pressure in a soft-extract state of preferably 25 to 35% by weight, but is not limited thereto.

In addition, the extract of the mixed herbal medicine prepared in step (a) to (c) may be further fractionated using one or more solvents selected from the group consisting of n-hexane, ethyl acetate and saturated n-butanol. This is not restrictive. In one embodiment of the present invention, the fractions of the extract include psychosaponin A, berberine and lycoisoflavones B, in particular, butanol and ethyl acetate fractions were confirmed to contain a high proportion of the components (Table 4). It was also confirmed that butanol and ethyl acetate fractions containing the above components had a high effect in gastrointestinal disease animal models (FIG. 3).

Thus, the composition prepared by the above method of the present invention comprises psychosaponin A, berberine and lysoisoflavone B, each containing 15 to 50 mg / g, 30 to 100 mg / g, and 0.5 mg / g to 5 mg / g, respectively. It can be used in the form of a pharmaceutical composition or food composition.

In another embodiment, the present invention is a gastrointestinal disease of 15 mg / g to 50 mg / g, berberine 30 mg / g to 100 mg / g and lysoisoflavone B 0.5 mg / g to 5 mg / g, or a composition comprising the same To provide a prophylactic or therapeutic use.

In another embodiment, the present invention is a gastrointestinal disease of 15 mg / g to 50 mg / g, berberine 30 mg / g to 100 mg / g and lysoisoflavone B 0.5 mg / g to 5 mg / g, or a composition comprising the same For the manufacture of a medicament for the prevention or treatment of cancer.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

Example 1 Preparation of Seahawks, Rhubarb and Licorice Extract-1

Shiho 260 kg, 60 kg of yellow lotus and 60 kg of licorice were extracted once with 48% of 2,660 L of 80% ethanol at 45 ° C. for 48 hours to obtain 2,000 kg of the extract, which was filtered through a filter press of 1 μm in size and then at 60 ° C. or lower and 500 mm Hg. The first step was concentrated under reduced pressure to obtain a 100 kg concentrate. After removing the water from the primary concentrate, the concentrate was dissolved in ethanol and concentrated again under reduced pressure to obtain a final soft extract 20kg.

Example 2: Preparation of Seahawks, Rhubarb and Licorice Extract-2

Shiho 373kg, rhubarb 84kg and licorice 84kg were extracted once with 2,660L of 80% ethanol at 45 ° C. for 91 hours to obtain 2,400 L of extract, which was filtered through a filter press of 1 μm size and then at 60 ° C. or lower, at 500 mmHg. The first step was concentrated under reduced pressure to obtain a 150 kg concentrate. The water of the first concentrate was removed, and then the concentrate was dissolved in ethanol and concentrated under reduced pressure again to obtain 40.6 kg of the final soft extract.

Example 3: Preparation of Seahawks, Rhubarb and Licorice Extract-3

Shiho 373kg, rhubarb 84kg and licorice 84kg were extracted once with 2,660L of 80% ethanol for 22 hours at 45 ° C. to obtain 2,000 kg of the extract, which was filtered through a filter press of 1 μm in size and then at 60 ° C. or below at 550 mmHg. The first step was concentrated under reduced pressure to obtain 119 kg concentrate. The water of the first concentrate was removed and the concentrate was dissolved in ethanol, and then concentrated under reduced pressure again to obtain a final soft extract 32.8 kg.

Example 4 Preparation of Seahawks, Rhubarb and Licorice Extract-4

Shiho 650kg, rhubarb 150kg and licorice 150kg were extracted once with 48% of 4,750L of 80% ethanol at 45 ° C. for 48 hours, and the extract filtrate was filtered under a filter press of 1 μm and concentrated under reduced pressure at 60 ° C. or lower to obtain 68 kg concentrate. . After removing the water from the primary concentrate, the concentrate was dissolved in ethanol, and then concentrated under reduced pressure again to obtain 74 kg of lead extract.

Example 5: Preparation of Fractions of Shiho, Rhododendron and Licorice Extracts

As shown in FIG. 1, 100 g of 80% ethanol extract of three kinds of sea tiger, rhubarb, and licorice extracted in Example 4 was suspended in an appropriate amount of 80% ethanol. Then, 300 g of silica gel was added, and the mixture was concentrated under reduced pressure on a water bath, and the mixture was packed into a column of 15 cm in diameter and 100 cm in height. After passing through 8L of n-hexane, 16L of ethyl acetate, 40L of saturated n-butanol, and concentrated under reduced pressure in each water bath to obtain a fraction for each solvent, the results are shown in Table 1 below.

Table 1

division	Mass (g)
Extract of Example 4	100
n-hexane fraction	0.687
Ethyl acetate fraction	19.5
Saturated n-butanol fraction	59
Non-soluble substance after saturation n-butanol fraction	2.5

Example 6: Preparation of Rectangle Tablets

75 g of the soft lead extract prepared in Example 4 and poloxamer and hydroxypropyl cellulose were dissolved in an appropriate amount of 80% ethanol, followed by stirring and mixing homogeneously. Separately, microcrystalline cellulose, pregelatinized starch, and colloidal silicon dioxide were mixed homogeneously, and then the prepared Shiho, yellow lotus and licorice ethanol solutions were added slowly to the mixture. Then, dried at 80 ℃ to produce granules of 30 mesh or more. Pre-gelatinized starch, croscarmellose sodium and magnesium stearate are added to the granules in a homogeneous manner, mixed homogeneously, and a rectangular tablet is prepared using a continuous tableting machine (Erweka, Germany) so that the total weight is 275 mg. Film coating in an appropriately sized film-coating apparatus.

Comparative Example 1: Preparation of Shiho, Rhododendron and Licorice Extract-5

By using the same method as the soft-ax extract of Examples 1 to 4 Shiho 60kg, 14kg, licorice 14kg and 80% ethanol using 430L Shiho, sulfur and licorice mixture was prepared to obtain about 6kg.

Comparative Example 2: Preparation of Shiho, Rhododendron and Licorice Extract-6

By using the same method as the soft-ax extract of Examples 1 to 4 Shiho, 10kg, rhubarb 2.3kg, licorice 2.3kg and 80% ethanol 73L to prepare a siho, sulfur and licorice mixture to obtain about 1kg.

Comparative Example 3: Preparation of Shiho, Rhododendron and Licorice Extract-7

By using the same method as the soft-ax extract of Examples 1 to 4 Shiho, 10kg, rhubarb 2.3kg, licorice 2.3kg and 80% ethanol 73L to prepare a siho, sulfur and licorice mixture to obtain about 1kg.

Experimental Example: Gastric mucosal protective effect of mixed herbal extracts including shiho, rhubarb and licorice and fractions thereof , and analysis of contents of berberine, psychosaponin A and lysoisoflavone B

Experimental Example 1-1. Confirmation of Gastric Mucosal Protective Effects of Mixed Herbal Extracts Containing Siho, Rhubarb and Licorice and Their Fractions Using Acute Gastritis Animal Model

Alcohol-induced acute gastritis animal model was used to confirm gastric mucosal protection, and the experiment was performed as follows.

Seven week old male rats were purchased and after one week of acclimatization, eight week old male rats (270g to 280g) were used for the experiment. Each group was divided into 8 groups, and each animal was orally administered with each extract at a dose of 48 hours under fast free water ingestion, and after 1 hour, rat ethanol (absolute ethanol) was weighed in rats. After oral administration of 5 mL / kg, gastric mucosal injury was induced for 1 hour. After 1 hour of anhydrous ethanol administration, rats were distal to the cervical spine and lethal, followed by extraction. The extracted stomach was infused with 2% formalin solution and fixed on ice for 1 hour. Then, the stomach was cut along the Taiwan section, spread out on a glass plate, and photographed. Gastric lesion area was measured using a photographic image to compare the degree of gastric mucosal injury to the group administered only with absolute ethanol alone.

Experimental Example 1-2. Content Analysis of Berberine, Psychosaponin A, and Lycoisoflavone B

The extracts or fractions prepared in Examples 1 to 5 and Comparative Examples 1 to 3 and the contents of berberine, psychosaponin A, and lycoisoflavone B of the drug product prepared in Example 6 were quantified under the following analysis conditions.

TABLE 2

System	Waters 2695 PDA 2996detectors
Column	shiseido capcell pak C18 250 * 4.6mm, 5㎛
Mobile phase	0.1% Trifluoroacetic acid: Acetonitrile (gradient system)
Running time	60min
Wavelength (nm)	254
Flow rate	1 mL / min
Sample	0.5g / 100mL

Experimental Example 1-3. Confirmation of gastric mucosal erosion effect of the composition comprising 15 mg / g to 50 mg / g of psychosaponin A, 30 mg / g to 100 mg / g of berberine and 0.5 mg / g to 5 mg / g of lysoisoflavone B

The present inventors confirmed the gastric mucosa protective effect of Shiho, rhubarb and licorice mixed herbal extract obtained in Example 4 and the organic solvent fraction obtained in Example 5.

As a result, when the lesion area by ethanol alone treatment was 100%, the fraction of Example 4 exhibited a value of 60.5%, resulting in a reduction of gastric mucosal damage, in particular the saturated n-butanol fraction and ethyl acetate. The fraction treated groups showed lesion areas of 55.5 and 54.6%, respectively, resulting in a significant reduction in gastric mucosal damage induced by ethanol. On the other hand, the hexane fraction showed a lesion area of 87.3%, which did not significantly reduce gastric mucosal damage to ethanol (FIG. 3).

As a result of performing component separation analysis by performing HPLC (High performance liquid chromatography) evaluation of the extract of Example 4 and the organic solvent fraction of Example 5, psychosaponin A (Saikosaponin A), psychosaponin B2 (Saikosaponin B2) , Saikosaponin D, Berberine, Palmintine, Palmitine, Coptisine, Liquiritin, Liquiritigenin, Licoisoflavone B, It was confirmed that it contains a substance such as ricochalcone A (Licochalcone A), quercetin (Quercetin), glycyrrhizin (Glycyrrhizin).

Said 13 substances were administered to the gastritis animal model of Experimental Example 1-1 at 10 mg / kg, respectively, and 3 substances psychosaponin A (Saikosaponin A) exhibiting gastric protective effect of 50% or more as shown in Table 3 below. , Berberine and Lycoisoflavone B were selected as key active ingredients.

TABLE 3

group	Gastric lesions (%)		% INH
	Mean	SD
Control	100.0	49.8	0.0
10 mg / kg Psychosaponin A	23	26.7	77.0
10mg / kg Lycoisoflavone B	48.8	45.5	51.2
10mg / kg berberine chloride	49.2	22.7	50.8
60 mg / kg Example 4	60.5	24.7	39.5

(% INH:% Inhibition)

Then, as a result of performing the HPLC analysis, it was confirmed that even in the purification of Example 5, the content of psychosaponin A (Saikosaponin A), berberine (Berberine), and lycoisoflavone B (Licoisoflavone B) was present in a predetermined amount or more. As a result of evaluating the contents present in the active ingredient layer, it was found that most of these components were transferred to the ethyl acetate fraction and the butanol fraction. In addition, the content ratio present in the active fraction layer relative to the total extract was as shown in Table 4.

Table 4

Classification	Saikosaponin A	Berberine	Licoisoflavone B
Ethyl acetate layer	One%	One%	0.03%
Buthanol layer	34%	47%	0%
Hexane layer	0%	0%	0%

Next, the gastritis protective effect of Examples 1 to 4 was confirmed using the gastritis pharmacological model of Example 1-1, and the HPLC content of each component present in the extract of each example was evaluated. The results are shown. As a result, Examples 1 to 4 showed gastritis inhibition of 56.0, 38.0, 43.0 and 39.5%, respectively.

Table 5

Classification	Saikosaponin A	Berberine	Licoisoflavone B	Drug efficacy (% INH)
Example 1	27.54 mg / g	65.44 mg / g	1.48mg / g	56.0
Example 2	21.8mg / g	45.69 mg / g	0.89 mg / g	38.0
Example 3	21.9 mg / g	43.32mg / g	0.68 mg / g	43.0
Example 4	28mg / g	60.39 mg / g	1.31 mg / g	39.5

(% INH:% Inhibition)

In addition, the degree of inclusion of each active ingredient in the tablet of Example 4 was confirmed and the results are shown in Table 6 below. Table 6 shows the amount present in tablet 1T, the amount of soft lead in tablet 1T is 150 mg, and the amount shown in Table 6 in mg / g is 28 mg / g for psychosaponin A, 60 mg / g for berberine, Lycoisoflavones B are converted to 1.2 mg / g.

Table 6

Classification	Saikosaponin A	Berberine	Licoisoflavone B
Tablet 1T of Example 4	4.2mg / T	9.0mg / T	0.18mg / T

In addition, the gastritis protective effect of Comparative Examples 1 to 3 was confirmed using the gastritis pharmacological model of Experimental Example 1-1, and the HPLC content of each component present in the extract of each comparative example was evaluated. The results are shown in Table 7 below.

TABLE 7

Classification	Saikosaponin A	Berberine	Licoisoflavone B	Drug efficacy (% INH)
Comparative Example 1	14.5mg / g	19.21 mg / g	N / A	23
Comparative Example 2	4.53mg / g	20.95 mg / g	N / A	13
Comparative Example 3	11.82mg / g	23.95 mg / g	N / A	-3

The above results indicate that the therapeutic effect against the excellent gastrointestinal diseases when the psychosaponin A, berberine and lysoisoflavone B of the present invention contains 15 to 50 mg / g, 30 to 100 mg / g and 0.5 mg / g to 5 mg / g, respectively. The result suggests that.

From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without changing the technical spirit or essential features. In this regard, the embodiments described above are to be understood in all respects as illustrative and not restrictive. The scope of the present invention should be construed that all changes or modifications derived from the meaning and scope of the following claims and equivalent concepts rather than the detailed description are included in the scope of the present invention.

Claims (14)

1. Saicosaponin A 15 mg / g to 50 mg / g, Berberine 30 mg / g to 100 mg / g and Licoisoflavone B (Licoisoflavone B) characterized in that it comprises 0.5 mg / g to 5 mg / g , Pharmaceutical composition for the prevention or treatment of gastrointestinal diseases.
2. The composition of claim 1, wherein the composition is a mixed herbal extract.
3. The composition of claim 1, wherein the composition further comprises a pharmaceutically acceptable carrier, excipient or diluent.
4. The gastrointestinal disease is gastritis, gastric ulcer, duodenal ulcer, Solinger-Elison syndrome, reflux esophagitis, postoperative ulcer, erosion of gastric mucosa, bleeding of gastric mucosa, redness of gastric mucosa and edema of gastric mucosa. The composition is selected from the group.
5. The composition of claim 1, wherein the gastrointestinal disease is mucosal damage of the stomach or duodenum caused by alcohol, smoking, stress, drugs, or a combination thereof.
6. According to claim 1, wherein the composition is a group consisting of tablets, pills, powders, granules, capsules, suspensions, liquid solutions, emulsions, syrups, sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations and suppositories A composition, characterized in that it has one of the formulations selected from.
7. Gastrointestinal disorders, including 15 mg / g to 50 mg / g of Saikosaponin A, 30 mg / g to 100 mg / g of Berberine and 0.5 mg / g to 5 mg / g of Lycoisoflavone B Food composition for prevention or improvement.
8. (a) water, 1 to 6 carbon atoms in a mixed herbal medicine comprising 10 parts by weight to 30 parts by weight of Cooptidis Rhizoma and 10 parts by weight to 30 parts by weight of licorice (Glycyrrhizae Radix) based on 100 parts by weight of Buleuri Radix Extracting with one or more solvents selected from the group consisting of alcohols and mixed solvents thereof;

   (b) concentration of the filtrate obtained in the extraction step under reduced pressure; And

   (c) suspending by adding alcohol to the reduced pressure concentrate of step (b), and then concentrating the secondary under reduced pressure to produce a soft extract.
9. The method of claim 8, wherein in the step (a), the solvent is 3 to 10 times solvent is used relative to the weight of the mixed herbal medicine.
10. The method of claim 8, wherein the method is characterized in that the extraction for 20 to 100 hours at an extraction temperature of 40 ℃ to 50 ℃, concentrated under reduced pressure at a temperature of 40 ℃ to 60 ℃.
11. The method of claim 8, wherein the first step (b) is concentrated under reduced pressure at a temperature of 500mmHg to 700mmHg and 40 ℃ to 60 ℃ dried to 10% by volume to 20% by volume.
12. The method of claim 8, wherein the second step of decompression concentration of step (c) is 25 to 35% by weight, characterized in that the drying under reduced pressure in the state of the soft extract.
13. The method of claim 8 further comprising (d) fractionating with one or more solvents selected from the group consisting of n-hexane, ethyl acetate and saturated n-butanol.
14. A method of treating gastrointestinal disease, comprising administering the pharmaceutical composition of claim 1 to a suspected gastrointestinal disorder.

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