KR20060125499A - Pharmaceutical composition comprising the catalpol derivatives isolated from the extract of pseudolysimachion longifolium having anti-inflammatory, anti-allergic and anti-asthmatic activity - Google Patents

Abstract

A pharmaceutical composition comprising the catalpol derivatives isolated from the extract of Pseudolysimachion longifolium having anti-inflammatory, anti-allergic and anti-asthmatic activity is provided to inhibit production of IgE(immunoglobulin E) in bronchoalveolar lavage fluid, and interleukin-4 and interleukin-13. The anti-inflammatory, anti-allergic and anti-asthmatic pharmaceutical composition comprises the catalpol derivatives represented by the formula(I) and isolated from the extract of Pseudolysimachion longifolium, or its pharmaceutically acceptable salt, wherein R is hydrogen atom, benzoyl group or cinnamoyl group independently substituted by at least one hydroxy C1-C3 lower alkyl group or lower alkoxy group. The anti-inflammatory, anti-allergic and anti-asthmatic health food comprises the catalpol derivatives represented by the formula(I) and sitologically acceptable additives, wherein the health food is formulated as tablet, capsule, pill or liquid.

Description

Pharmaceutical composition comprising the catalpol derivatives isolated from the extract of Pseudolysimachion longifolium having anti-inflammatory, anti-allergic and anti-asthmatic activity}

1 is a diagram showing airway hypersensitivity as a penh value by recording the respiratory rate due to bronchial contraction using a body-body plethysmography (A), with egg white albumin In the case of airway sensitization and secondary antigen administration (B), berproside administration of secondary antigen administration (C), and montelukast administration of secondary antigen administration (D). ,

Figure 2 shows the analysis of the histology of bronchoalveolar alveolar, when airway sensitization (I), airway sensitization and second antigen administration with egg white albumin (II), berproside at the second antigen administration In the case of administration of (III) and the administration of montelukast when the second antigen was administered (Ⅳ), the effect on the bronchial tissue cells and the degree of inflammatory cell influx.

Figure 3 shows the secretion of mucus in the endothelial cells of the bronchoalveolar, when airway sensitization (I), airway sensitization and second antigen administration with egg white albumin (II), berproside during second antigen administration In the case of administration of (III), when montelukast was administered during the second antigen administration (IV), the mucous membranes of bronchial tissue endothelial cells were stained purple to observe the amount of secretion of mucus.

The present invention relates to a pharmaceutical composition containing a catalpol derivative isolated from Pseudolysimachion longifolium extract having anti-asthmatic activity as an active ingredient.

Asthma is a disease characterized by airway hypersensitivity to various stimuli. Clinical symptoms such as wheezing, dyspnea, and cough caused by extensive airway narrowing are reversibly improved naturally or by treatment. Can be. Most asthma are allergic and characterized by chronic airway inflammation and bronchial hyperresponsiveness (Minoguchi K and Adachi M. Pathophysiology of asthma.In: Cherniack NS, Altose MD, Homma I, editors Rehabilitation of the patient with respiratory disease.New York: McGraw-Hill , 1999, pp97-104).

Allergic asthma patients are known to account for about 10% of the world's population (275 million, 1995), and 17 million people in the United States alone suffer from asthma, of which 5 million are reported to be adolescents. In 2000, the US market for allergic asthma drugs was about $ 6.4 billion, and the Korean market was reported to be about $ 1 billion, accounting for 20% of the total Korean drug market.

Asthma can be divided into exogenous and endogenous asthma depending on the cause. In exogenous asthma, it is asthma that causes symptoms when exposed to the causative antigen. A skin test or bronchial challenge test for the causative antigen is positive and usually occurs young. House dust and mites are the most common causative agents, and pollen, animal epithelium, and fungus act as causative antigens. In the case of endogenous asthma, upper respiratory tract infections, exercise, emotional instability, cold climate and humidity changes cause or worsen asthma, which is common in adult-type asthma. Other medications include asthma, exercise-induced asthma and occupational asthma. In the case of the most problematic exogenous asthma, the causative antigen can be identified by skin test. In exogenous asthma, the total IgE in serum is increased and antigen specific IgE is detected.

In terms of pathophysiology, asthma is recognized as a chronic inflammatory disease caused by the proliferation, differentiation and activation of inflammatory cells by cytokines produced by TH2 immune cells, which migrate and invade airways and surrounding airways (Elias JA, Lee CG). , Zheng T, Ma B, Homer RJ, Zhu Z. New insights into the pathogenesis of Asthma., J. Clin.Invest . , 111 , pp291-297, 2003). In this case, inflammatory cells such as activated eosinophils, mast cells, and alveolar macrophages secrete various inflammatory mediators, which are involved in the production of cytokines such as IL-4, IL-5, and IL-13, which are involved in inflammatory cell activation. Cysteine leukotriene biosynthesis secreted from inflammatory cells such as eosinophils is the main cause of inflammation and allergic reactions and asthma due to their action, and many studies have been conducted to develop drugs for inhibiting their production.

Pseudolysimachion longifolium is a perennial herb of dicotyledon plywood group currency plant phenomena. It is a perennial herb distributed in Korea, China, Russia, and Europe. Aglycon is 6-hydroxyluteolin (Information and Shin Mingyo, Pharmacokinetic Dictionary, pp913-914, 1998).

The present inventors have found that the catalpol derivatives isolated from Rhizome oleracea extract induce an airway hypersensitivity, as well as immunoglobulin E (IgE) and interleukin-4 (IL-4), interleukin-13 in bronchial alveolar fluid, in an asthma model in which ovarian albumin was sensitized. The present invention was completed by identifying the activity of inhibiting the production of (IL-13), the activity of inhibiting eosinophil increase, and the activity of inhibiting edema in a carrageenan-induced animal model.

The present invention is to provide a pharmaceutical composition and health functional food for the prevention or treatment of inflammatory diseases, allergies and asthma, containing a catalpol derivative having anti-inflammatory, anti-allergic and anti-asthmatic activity as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention or treatment of inflammation, allergies and asthma diseases, including the catalpol derivative of the general formula (I) or a pharmaceutically acceptable salt thereof .

(Ｉ)

(I)

In the above, R is a benzoyl group each independently substituted with a hydrogen atom or at least one hydroxy C ₁ -C ₃ lower alkyl group or a lower alkoxy group.

As a preferable compound group of the said General formula (I), R substituent is 3, 4- dihydroxy benzoyl (3, 4- dihydroxy benzoyl), 4-hydroxy-3- methoxy benzoyl (4-hydroxy-3-methozybenzoyl) , 3-hydroxy-4-methoxybenzoyl3-hydroxy-4-methozybenzoyl), 4-hydroxybenzoyl, 3,4-dimethoxybenzoyl (3,4-dimethoxybenzoyl) 3,4-di Hydroxycinnamoyl (3,4-dihydroxycinnamoyl) and 3-hydroxy-4-methoxycinnamoyl (3-hydroxy-4-methoxycinnamoyl).

Hereinafter, the present invention will be described in more detail.

The catalpol derivative compound of the general formula (I) of the present invention may be synthesized through the synthesis and fractionation of conventional substituents by separating from Ginsan oleacea extract or known synthetic methods well known in the art (Herbert). O. House: Modern Synthetic Reactions , 2nd Ed ., The Benjamin / Cummings Publishing Co., 1972).

Specifically looking at the process of separating the catalpol derivative compound of the present invention from the extract of Ginsan olecus, Ginsan oleacea crude extract of the present invention after collecting, drying and crushing Ginsan oleus, powdered, about 2 of the weight of Ginsan oleus sample weight about one of C ₁ to C ₄ lower alcohol or a polar solvent of these, such as to 20 times by volume up to the water and methanol, ethanol, butanol, a mixed solvent having a mixing ratio of 0.1 to 1: 10, preferably methanol 2 to 5 times, preferably, using an extraction method such as hot water extraction, cold needle extraction, reflux cooling extraction or ultrasonic extraction at 20 to 50 ° C. for about 10 hours to 48 days, preferably 20 hours to 30 hours. After 2-3 extractions, it can be obtained by filtration, concentration and drying according to conventional methods.

After the crude extract is suspended in distilled water, the suspension is added 1 to 10 times, preferably by adding about 1 to 100 times the volume, preferably about 1 to 5 times the volume of a polar solvent such as ethanol, methanol and butanol. 1 to 5 times the polar solvent soluble layer is extracted and separated to obtain a soluble fusiol butanol extract, chloroform-methanol mixture step by step to perform silica gel column chromatography to obtain 6 fractions, the bottom fraction 3 was added chloroform-methanol mixture and subjected to silica gel column chromatography again to obtain 12 fractions. Again, thin layer chromatography and reverse phase 18-column chromatography were performed sequentially to separate the main components and subjected to Sephadex LH-20 column chromatography to obtain a cataful derivative compound of the present invention.

The present invention provides a pharmaceutical composition for the prevention and treatment of anti-inflammatory, anti-allergic and asthma diseases, containing the catapult derivative compound obtained by the above method as an active ingredient.

The catalpol derivative compounds of formula (I) may be prepared with pharmaceutically acceptable salts and solvates according to methods conventional in the art.

As the pharmaceutically acceptable salt, acid addition salts formed by free acid are useful. Acid addition salts are prepared by conventional methods, for example by dissolving a compound in an excess of aqueous acid solution and precipitating the salt using a water miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. Equal molar amounts of the compound and acid or alcohol (eg, glycol monomethyl ether) in water can be heated and the mixture can then be evaporated to dryness or the precipitated salts can be suction filtered.

In this case, organic acids and inorganic acids may be used as the free acid, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, etc. may be used as the inorganic acid, and methanesulfonic acid, p -toluenesulfonic acid, acetic acid, trifluoroacetic acid, Citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid ( gluconic acid), galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanic acid, hydroiodic acid, and the like.

Bases can also be used to make pharmaceutically acceptable metal salts. An alkali metal or alkaline earth metal salt is obtained by, for example, dissolving a compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and then evaporating and drying the filtrate. At this time, as the metal salt, it is particularly suitable to prepare sodium, potassium or calcium salt, and the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

Pharmaceutically acceptable salts of these compounds include salts of acidic or basic groups which may be present in the tricine compound, unless otherwise indicated. For example, pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulphate, phosphate, hydrogen phosphate, dihydrogen Phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p -toluenesulfonate (tosylate) salts, and methods or processes for preparing salts known in the art It can be prepared through.

In addition, Ginsan tail grass is a drug that has been used for a long time as a herbal medicine and edible compounds of the present invention extracted from them also have no problems such as toxicity and side effects.

As a result of confirming the efficacy of anti-inflammatory, anti-allergic and anti-asthmatic activity of the catalpol derivative compounds of the present invention, airway hypersensitivity inhibitory activity, bronchial alveolar fluid in the asthma animal model sensitized with egg white albumin It was confirmed that -4 (IL-4), interleukin-13 (IL-13) activity inhibits the production, eosinophil increase activity and carrageenan-induced animal model edema inhibition activity.

The pharmaceutical composition comprising the compound of the present invention comprises 0.1 to 50% by weight of the extract or compound based on the total weight of the composition.

Pharmaceutical compositions comprising the compounds of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Pharmaceutical dosage forms of the compounds of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.

The pharmaceutical compositions comprising the compounds according to the present invention may be prepared in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Can be formulated and used. Carriers, excipients and diluents that may be included in the composition comprising the compound include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate, sucrose ( It is prepared by mixing sucrose or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the compounds of the present invention depend on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, but may be appropriately selected by those skilled in the art. However, for the desired effect, the compound of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably at 0.001 to 10 mg / kg. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

The compounds of the present invention can be administered to mammals such as mice, mice, livestock, humans, and the like by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

The present invention provides a dietary supplement comprising the compound having anti-inflammatory, anti-allergic and anti-asthmatic activity, and a food acceptable additive. Examples of the food to which the compound of the present invention can be added include various foods, beverages, gums, teas, vitamin complexes, and health functional foods.

It may also be added to foods or beverages for the purpose of preventing anti-inflammatory, anti-allergic and asthmatic diseases. At this time, the amount of the extract in the food or beverage may be added in 0.01 to 15% by weight of the total food weight, the health beverage composition may be added in a ratio of 0.02 to 5 g, preferably 0.3 to 1g based on 100 ml. have.

Health functional food of the present invention includes the form of tablets, capsules, pills, liquids and the like.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for containing the compound as an essential ingredient in the indicated ratios, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the compounds of the present invention include various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the compounds of the present invention may contain flesh for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is usually selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the compound of the present invention.

Below, The invention is illustrated in detail by the following examples and experimental examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following Examples and Experimental Examples.

Example 1 Isolation and Identification of Catalpol Derivatives

1-1. Preparation of Crude Extracts of Kinsan Oyster Grass

7.9 kg of Ginsan Oyster Outpost was dried and pulverized, 50 L of methanol was added thereto, stirred at room temperature for 24 hours, and the supernatant was recovered by vacuum filtration. This process was repeated three times to collect the supernatant, and then concentrated under reduced pressure to obtain 950.5 g of a crude methanol extract of ginsan coriander.

1-2. Isolation and Identification of Catalpol Derivatives

The crude extract of Ginsan coriander obtained in Example 1-1 was adsorbed onto silica gel, and subjected to silica gel column chromatography (72-230 mesh, 8.5 × 65 cm) in two portions to sequentially polarize the hexane-chloroform-methanol mixed solvent. Elution with eluting gave 5 fractions. Using the LTC ₄ synthetic assay, the most active fractions 4 and 5 were selected and mixed, and then 425 g were suspended in 10 L of distilled water, and 10 L of ethyl acetate and 10 L of butanol were sequentially added to the ethyl acetate soluble fraction. The butanol soluble fraction and the water soluble fraction were separated. The butanol soluble fraction was filtered and concentrated under reduced pressure to remove the solvent, and 144.0 g of gluconate butanol soluble extract was subjected to reverse phase silica gel chromatography (72-230 mesh, 8.5 × 65 cm) (methanol 0%, 10%, 30). Stepped to%, 50% and 100%) to give five fractions. The second fraction (29.1 g) was subjected to silica gel column chromatography using a chloroform-methanol mixture (ratio of 10/1-1/1) to separate 7 fractions (2-1 to 2-7). The -4 fraction was recrystallized in methanol to give 14.2 g of verproside (6-O-3,4-Dihydroxybenzoyl catalpol) having spectroscopic data as follows.

In the third fraction (17.3 g), silica gel column chromatography was performed with a chloroform-methanol mixture (ratio of 10/1-1/1) to obtain seven fractions (3-1 to 3-7). The 3-3 fraction (8.5 g) was recrystallized in methanol to obtain 7.2 g of isovanillyl catalpol (6-O-3-hydroxy-4-methoxybenzoly catalpol) having spectroscopic data as follows. It was. In addition, 3-5 fractions (1.5 g) obtained above were subjected to reverse phase-18 column chromatography (methanol: water = 1: 4), followed by separation of the main components, followed by Sephadex LH-20 column chromatography (methanol: water = 85:15), picroside II (6-O-4-hydroxy-3-methozybenzoyl) and verminoside (verminoside, 6-O-3,4-) having the following spectroscopic data dihydroxycinnamoyl catalpol) was obtained at 101.0 mg and 30.0 mg, respectively.

In the fourth fraction (6.2 g) 15 fractions (4-1 to 4-15) were obtained. 1.2 g of the 4-3 fraction obtained above was recrystallized in methanol to have 6-O-veratroylcatalpol (6-O-veratroylcatalpol, 6-O-3,4-Dimethoxybenzoyl) having the following spectroscopic data. Mg was obtained. In addition, 4-4 fractions (261.0 mg) and 4-5 fractions (288.0 mg) were combined, and silica gel column chromatography was again performed with a chloroform-methanol mixture (ratio of 10/1-5/1). 52.5 mg of minecoside with data (minecoside, 6-O-3-hydroxy-4-methozycinnamoyl catalpol) was obtained.

On the other hand, the hydrolysis of the above-mentioned verproside to obtain a catalpol (compound 1), a common substructure of compounds 2 to 7, 109 mg of compound 2 was added to 0.1N KOH methanol solution 8 at room temperature After stirring for a period of time, neutralized with 0.1 N hydrochloric acid solution and the reaction was concentrated under reduced pressure to carry out reverse phase-18 column chromatography (methanol: water = 1: 4) to give 54.0 mg of catalpol having the following spectroscopic data. .

Catalpol

¹ H-NMR (400 MHz, DMSO- d ₆ ) δ: 2.12 (1H, dddd , J = 1.6, 4.0, 8.0, 8.0 Hz, H-5), 2.31 (1H, d , J = 8.0, 9.6 Hz, H-9), 3.00 (1H, m , H-G4), 3.05 (1H, m , H-G2), 3.11 (1H, m , H-G5), 3.17 (1H, m , H-G3), 3 , 34 (1H, br s , H-7), 3.40, 3.70 (2H, m , H-G6), 3.63, 3.87 (2H, d , J = 12.8, each, H-10), 3.76 (1H, d , J = 8.0 Hz, H-6), 4.59 (1H, d , J = 8.0 Hz, H-G1), 4.90 (1H, d , J = 9.6 Hz, H-1), 5.01 (1H, dd , J = 4.6, 6.0 Hz, H-4), 6.36 (1H, dd , J = 1.6, 6.0 Hz, H-3).

¹³ C-NMR (100 MHz, DMSO- d ₆ ) δ: 93.2 (C-1), 140.2 (C-3), 103.3 (C-4), 37.4 (C-5), 77.1 (C-6), 60.7 (C-7), 64.8 (C-8), 42.1 (C-9), 58.9 (C-10), 97.8 (C-G1), 73.4 (C-G2), 76.4 (C-G3), 70.2 (C-G4), 77.4 (C-G5), 61.3 (C-G6).

6-O- (3,4-dihydroxybenzoyl) catapol (verproside)

¹ H NMR (400 MHz, DMSO- d ₆ ) δ: 2.47 (1H, dd , J = 8.0, 9.2 Hz, H-9), 2.59 (1H, dddd , J = 1.6, 4.0, 8.0, 8.0, H- 5), 3.00 (1H, m , H-G4), 3.05 (1H, m , H-G2), 3.14 (1H, m , H-G5), 3.18 (1H, m , H-G3), 3.42, 3.71 (2H, m , H-G6). 3.67 (1H, s , H-7), 3.71, 3.91 (2H, d , J = 13.2 Hz, each, H-10), 4.61 (1H, d , J = 7.6 Hz, H-G1), 4.94 (1H , dd , J = 4.0, 6.0 Hz, H-4), 5.03 (1H, d , J = 8.0 Hz, H-6), 5.09 (1H, d , J = 9.2 Hz, H-1), 6.41 (1H , dd , J = 1.6.6.0 Hz, H-3), 6.82 (1H, d , J = 8.0 Hz, H-5 '), 7.35 (1H, dd , J = 2.0, 8.0 Hz, H-6') , 7.39 (1H, doublet , J = 2.0 Hz, H-2 ').

¹³ C-NMR (100 MHz, DMSO- d ₆ ) δ: 93.0 (C-1), 141.1 (C-3), 101.8 (C-4), 35.2 (C-5), 79.5 (C-6), 58.2 (C-7), 65.8 (C-8), 41.8 (C-9), 120.0 (C-1 '), 116.4 (C-2'), 145.1 (C-3 '), 150.8 (C-4 '), 115.4 (C-5'), 122.6 (C-6 '), 165.6 (C-7'), 97.9 (C-G1), 73.4 (C-G2), 76.4 (C-G3), 70.3 ( C-G4), 77.5 (C-G5), 61.4 (C-G6).

6-O- (4-hydroxy-3-methoxybenzoyl) catapol (picroside II)

¹ H-NMR (400 MHz, DMSO- d ₆ ) δ: 2.47 (1H, dd , J = 8.0, 9.6 Hz, H-9), 2.58 (1H, dddd , J = 1.2, 6.0, 8.0, 8.4 Hz, H-5), 3.00 (1H, m , H-G4), 3.05 (1H, m , H-G2), 3.14 (1H, m , H-G5), 3.18 (1H, m , H-G3), 3.42 , 3.71 (2H, m , H-G6), 3.67 (1H, br s , H-7), 3.72, 3.92 (2H, d , J = 13.2, each, H-10), 4.62 (1H, d , J = 7.6 Hz, H-G1), 4.99 (1H, dd , J = 4.4, 6.0 Hz, H-4), 5.06 (1H, d , J = 8.4 Hz, H-6), 5.11 (1H, d , J = 9.6 Hz, H-1), 6.42 (1H, dd , J = 1.2.6.0 Hz, H-3), 6.89 (1H, d , J = 8.4 Hz, H-5 '), 7.46 (1H, d , J = 2.0 Hz, H-2 '), 7.52 (1H, dd , J = 2.0, 8.4 Hz, H-6'), 3.83 (3H, s , 3'-0-CH ₃ ).

¹³ C-NMR (100 MHz, DMSO- d ₆ ) δ: 93.0 (C-1), 141.1 (C-3), 101.8 (C-4), 35.2 (C-5), 79.7 (C-6), 58.2 (C-7), 65.8 (C-8), 41.8 (C-9), 58.5 (C-10), 120.0 (C-1 '), 112.7 (C-2'), 147.5 (C-3 ' ), 152.0 (C-4 '), 115.3 (C-5'), 123.8 (C-6 '), 165.6 (C-7'), 97.9 (C-G1), 73.4 (C-G2), 76.4 ( C-G3), 70.3 (C-G4), 77.5 (C-G5), 61.4 (C-G6), 55.7 (3'-OCH ₃ ).

6-O- (3-hydroxy-4-methoxybenzoyl) catapol (isovanylyl catapol)

¹ H-NMR (400 MHz, DMSO- d ₆ ) δ: 2.47 (1H, m , H-9), 2.55 (1H, m H-5), 3.00 (1H, m , H-G4), 3.05 (1H , m , H-G2), 3.14 (1H, m , H-G5), 3.18 (1H, m , H-G3), 3.43, 3.70 (2H, m , H-G6), 3.70 (1H, br s , H-7), 3.72, 3.92 (2H, d , J = 13.2, each, H-10), 4.62 (1H, d , J = 8.0 Hz, H-G1), 4.95 (1H, dd , J = 4.4, 6.0 Hz, H-4), 5.06 (1H, d , J = 8.0 Hz, H-6), 5.11 (1H, d , J = 9.2 Hz, H-1), 6.42 (1H, d , J = 6.0 Hz , H-3), 7.04 (1H, d , J = 8.4 Hz, H-5 '), 7.42 (1H, br s , H-2'), 7.48 (1H, d , J = 8.4 Hz, H-6 '), 3.84 (3H, s , 4'-0-CH ₃ ).

¹³ C-NMR (100 MHz, DMSO- d ₆ ) δ: 93.0 (C-1), 141.0 (C-3), 101.6 (C-4), 35.2 (C-5), 79.7 (C-6), 58.2 (C-7), 65.8 (C-8), 41.8 (C-9), 58.4 (C-10), 121.7 (C-1 '), 115.7 (C-2'), 146.3 (C-3 ' ), 152.1 (C-4 '), 111.4 (C-5'), 121.3 (C-6 '), 165.3 (C-7'), 97.8 (C-G1), 73.4 (C-G2), 76.4 ( C-G3), 70.3 (C-G4), 77.4 (C-G5), 61.4 (C-G6), 55.7 (4'-OCH ₃ ).

6-O- (4-hydroxybenzoyl) catalpol (catalfoside)

¹ H-NMR (400 MHz, DMSO- d ₆ ) δ: 2.47 (1H, dd , J = 8.0, 9.6 Hz, H-9), 2.56 (1H, dddd , J = 1.2, 4.0, 8.0, 8.0 Hz, H-5), 3.00 (1H, m , H-G4), 3.05 (1H, m , H-G2), 3.14 (1H, m , H-G5), 3.18 (1H, m , H-G3), 3.43 , 3.72 (2H, m , H-G6), 3.69 (1H, br s , H-7), 3.72, 3.92 (2H, d , J = 13.2 Hz, each, H-10), 4.62 (1H, d , J = 8.0 Hz, H-G1), 4.96 (1H, dd , J = 4.0, 6.0 Hz, H-4), 5.05 (1H, dd , J = 1,2, 8.0 Hz, H-6), 5.11 ( 1H, d , J = 9.6 Hz, H-1), 6.42 (1H, dd , J = 1.2.6.0 Hz, H-3), 6.86 (2H, d , J = 8.0 Hz, H-3 ', -5 '), 7.85 (2H, d , J = 2.0 Hz, H-2', -6 ').

¹³ C-NMR (100 MHz, DMSO- d ₆ ) δ: 92.9 (C-1), 141.1 (C-3), 101.8 (C-4), 35.1 (C-5), 79.6 (C-6), 58.2 (C-7), 65.8 (C-8), 41.8 (C-9), 119.6 (C-1 '), 131.7 (C-2', 6 '), 115.5 (C-3', 5 ') , 162.6 (C-4 '), 165.5 (C-7'), 97.8 (C-G1), 73.4 (C-G2), 76.4 (C-G3), 70.3 (C-G4), 77.5 (C-G5 ), 61.4 (C-G6).

6-O- (3,4-dimethoxybenzoyl) catalpol (6-O-veratroylcatalpol)

¹ H-NMR (400 MHz, DMSO- d ₆ ) δ: 2.47 (1H, dd, J = 8.0, 9.6 Hz, H-9), 2.59 (1H, dddd , J = 1.6, 4.8, 8.0,8.0 Hz, H-5), 3.00 (1H, m , H-G4), 3.05 (1H, m , H-G2), 3.14 (1H, m , H-G5), 3.18 (1H, m , H-G3), 3.42 , 3.71 (2H, m , H-G6), 3.70 (1H, br s , H-7), 3.72, 3.90 (2H, d , J = 13.2 Hz, each, H-10), 4.61 (1H, d , J = 7.6 Hz, H-G1), 4.97 (1H, dd , J = 4.8, 6.0 Hz, H-4), 5.08 (1H, d , J = 8.8 Hz, H-6), 5.10 (1H, d , J = 9.6 Hz, H-1), 6.42 (1H, dd , J = 1.6.6.0 Hz, H-3), 7.09 (1H, d , J = 8.4 Hz, H-5 '), 7.46 (1H, d , J = 2.0 Hz, H-2 ′), 7.64 (1H, dd , J = 2.0, 8.4 Hz, H-6 ′), 3.81, 3.84 (6H, s each, 3 ′, 4′-0CH ₃ ).

¹³ C-NMR (100 MHz, DMSO- d ₆ ) δ: 92.9 (C-1), 141.1 (C-3), 101.8 (C-4), 35.2 (C-5), 79.9 (C-6), 58.2 (C-7), 65.9 (C-8), 41.8 (C-9), 58.4 (C-10), 121.3 (C-1 '), 111.8 (C-2'), 148.5 (C-3 ' ), 153.2 (C-4 '), 111.2 (C-5'), 123.5 (C-6 '), 165.5 (C-7'), 97.8 (C-G1), 73.4 (C-G2), 76.4 ( C-G3), 70.3 (C-G4), 77.5 (C-G5), 61.4 (C-G6), 55.6, 55.7 (3 ', 4'-OCH ₃ ).

6-O- (3,4-dihydroxycinnamoyl) catalpol (verminoside)

¹ H-NMR (400 MHz, DMSO- d ₆ ) δ: 2.43 (1H, m, H-9), 2.45 (1H, m, H-5), 3.01 (1H, m , H-G4), 3.05 ( 1H, m , H-G2), 3.14 (1H, m , H-G5), 3.18 (1H, m , H-G3), 3.42, 3.70 (2H, m , H-G6), 3.64 (1H, br s , H-7), 3.71, 3.90 (2H, d , J = 13.2 Hz, each, H-10), 4.61 (1H, d , J = 8.4 Hz, H-G1), 4.94 (1H, dd , J = 4.0, 5.6 Hz, H-4), 4.99 (1H, d , J = 7.2 Hz, H-6), 5.08 (1H, d , J = 9.2 Hz, H-1), 6.42 (1H, d , J = 5.6 Hz, H-3), 6.77 (1H, d , J = 8.0 Hz, H-5 '), 7.08 (1H, d , J = 1.6 Hz, H-2'), 7.05 (1H, dd , J = 1.6, 8.0 Hz, H-6 ').

¹³ C-NMR (100 MHz, DMSO- d ₆ ) δ: 92.9 (C-1), 141.1 (C-3), 101.7 (C-4), 35.1 (C-5), 79.2 (C-6), 58.2 (C-7), 65.7 (C-8), 41.8 (C-9), 58.5 (C-10), 125.4 (C-1 '), 115.8 (C-2'), 146.0 (C-3 ' ), 148.6 (C-4 '), 113.3 (C-5'), 121.6 (C-6 '), 145.6 (C-7'), 115.0 (C-8 '), 97.9 (C-G1), 73.4 (C-G2), 76.4 (C-G3), 70.3 (C-G4), 77.5 (C-G5), 61.4 (C-G6).

6-O- (3-hydroxy-4-methoxycinnamoyl) catalpol (minecoside)

¹ H-NMR (400 MHz, DMSO- d ₆ ) δ: 2.46 (1H, m, H-9), 2.48 (1H, m, H-5), 3.00 (1H, m , H-G4), 3.05 ( 1H, m , H-G2), 3.14 (1H, m , H-G5), 3.18 (1H, m , H-G3), 3.42, 3.70 (2H, m , H-G6), 3.67 (1H, br s , H-7), 3.72, 3.91 (2H, d , J = 13.2 Hz, each, H-10), 4.61 (1H, d , J = 8.8 Hz, H-G1), 4.94 (1H, dd , J = 4.0, 6.0 Hz, H-4), 5.00 (1H, d , J = 7.2 Hz, H-6), 5.09 (1H, d , J = 9.2 Hz, H-1), 6.42 (1H, dd , J = 1,2, 5.6 Hz, H-3), 6.96 (1H, d , J = 8.0 Hz, H-5 '), 7.13 (1H, d , J = 2.0 Hz, H-2'), 7.17 (1H, dd , J = 2.0, 8.0 Hz, H-6 '), 3.82 (3H, s, -OCH ₃ ).

¹³ C-NMR (100 MHz, DMSO- d ₆ ) δ: 93.0 (C-1), 141.1 (C-3), 101.7 (C-4), 35.1 (C-5), 79.3 (C-6), 58.2 (C-7), 65.7 (C-8), 41.8 (C-9), 58.5 (C-10), 126.8 (C-1 '), 114.5 (C-2'), 146.7 (C-3 ' ), 150.2 (C-4 '), 112.0 (C-5'), 121.4 (C-6 '), 145.7 (C-7'), 114.5 (C-8 '), 97.9 (C-G1), 73.4 (C-G2), 76.4 (C-G3), 70.3 (C-G4), 77.5 (C-G5), 61.4 (C-G6), 55.6 (4'-OCH ₃ ).

Reference Example 1. Cell Viability Experiment

Cell viability test was performed by standard MTT method using promyelotic HL-60 cell line (HC-18103, Korea Gene Bank) (Wang Z, Zhou J, Ju Y, Zhang H, Liu M, Li) Z., Effects of two saponins extracted from the Polygonatum Zanlanscianense pamp on the human leukemia (HL-60) cells.Biol ., Pharm. Bull. , 24 , pp159-162, 2001). The cells were diluted at 5 × 10 ⁵ cells / ml in IMCOve's Modified Dulbecco's Medium (IMDM) medium (GibcoBRL) containing 5% fetal bovine serume, 100 U / ml penicillin and 100 μg / ml streptomycin. Afterwards, the cells were incubated for 24 hours at 37 ° C. in 96-microtiter wells. Thereafter, 10 μl of a catalpol derivative sample dissolved in DMSO and 10 μl of MTT (5 mg / ml) dissolved in PBS were added thereto, and the mixture was incubated for 4 hours under the same culture conditions, and the mixture was reacted at 3,000 rpm to obtain a formazan precipitate in the reaction solution. Centrifuge for minutes. The precipitate obtained after centrifugation was dissolved in 100 μl of DMSO and measured for absorbance at 570 nm using a microplate reader (BIO-RAD, USA) as a percentage of the control group that was not administered the quincetail extract of the present invention. Cell viability was calculated.

As shown in Table 1, the cell viability of the catalpol derivatives extracted from Gansan Koji was measured 98% -116% at 50μM and 95% -114% at 100μM, indicating no cytotoxicity. .

 sample Cell survival rate (%)    50 μM    100 μM  Verfroside  105  102  6-O-Veratroyl Catalyst  116  114  Minecoside  98  95

Reference Example 2 Airway Sensitization and Antigen Administration of Laboratory Animals

8-week-old pathogen-uninfected white rat female (Balb / c) (weight: approx. 20 g) After purchase from Orient (Seoul, Korea), 20 mg of 2 mg aluminum hydroxide (Sigma A8222) and egg white albumin at 2 week intervals (Sigma A5503) was sensitized by injecting 100 [mu] l of suspended phosphate buffer solution (pH 7.4) into the abdominal cavity twice. After 28, 29, and 30 days, a phosphate buffer solution containing 1% egg white albumin was sprayed for 20 minutes using an ultrasonic nebulizer, and a negative control group (5 mice) did not cause airway sensitization. After sensitization PBS 50 mg / kg administration group (5), 30 mg / kg berproside obtained in Example 1 and 30 mg / kg of montelukast obtained in Example 1 as an experimental group in the phosphate buffer solution to administer the antigen 1 The experiment was performed by oral administration before time.

Experimental Example 1. Analysis of airway hypersensitivity lowering effect

Airway hypersensitivity was measured 24 hours after the last challenge in Reference Example 2. Measures of airway hypersensitivity were measured by spraying methacholine solutions at concentrations of 0, 12.5, 25 and 50 mg / ml using whole-body plethysmography, and then recording the respiratory rate due to bronchial contraction as a percentage in Penh values. Converted to. As shown in FIG. 1, berproside showed a significant decrease in% Penh value in methacholine 25 and 50 mg / kg treatment groups compared to OVA-treated animals, effectively reducing airway hyperresponsiveness. .

Experimental Example 2 Analysis of Inflammatory Mediators of Plasma and Bronchoalveolar Fluids

2-1. Immunoglobulin E (IgE) Analysis of Plasma and Bronchial Alveoli (BALF)

After carrying out Experimental Example 1 and administering an excess pentobarbital (Sigma P3761), the plasma was obtained from blood collected from the heart from each group, and a cannula was inserted into the trachea after bronchial incision. Bronchoalveolar fluid (BALF) was obtained by inhaling 0.5 ml three times by the method. IgE content was analyzed according to the manufacturer's method using an IgE antibody ELISA kit (PharMingen, San Diego, Calif., USA) to determine the content of IgE in bronchial alveoli. The concentration of IgE was calculated by diluting the standard (52,000 ng / ml) step by step and measuring the absorbance at 405 nm to prepare the absorbance standard for each concentration, and then calculated the concentration by converting the absorbance of the bronchial alveolar fluid.

As a result, as shown in Table 2, IgE production increased rapidly in the animal group sensitized with OVA, and it was confirmed that BERPROSID effectively suppressed IgE production.

Inhibitory Activity of IgE Production in Plasma and Bronchoalveolar Fluid of Verproside Kinds  IgE (ng / ml)  BALF  plasma  (-) Control  1.0 ± 0.018  16.1 ± 23.9  OVA treatment group 59.4 ± 35.5 ^* 85.6 ± 17.0 ^*  OVA + Verphoxide (30mg / kg) treated group (% inhibition) 21.5 ± 11.2 ^** (63.8%) 40.2 ± 13.2 ^** (53.0%)  OVA + montelukast (30mg / kg) treated group (% inhibition) 3.8 ± 0.7 ^** (93.6%) 31.4 ± 14.2 ^** (63.3%)  * significant difference from normal control (NC) group p <0.005 ** significant difference from OVA treated group, p <0.05

2-2. Bronchus Alveolar fluid (BALF)  Cytokine Analysis

IL-4 and IL-13 specific ELISA kits (R & D Systems, Minneapolis, USA) were used to measure the cytokine content of 50 μl of the bronchial alveoli of each experimental zone obtained in Experimental Example 2-1. According to the method, the content of each cytokine was measured.

As a result of the experiment, as shown in Table 3, the production of 1L-4 and IL-13 was significantly increased in the experimental group sensitized with egg white albumin (OVA) compared to the untreated group, at which time OVA + Verproside 30 mg In the experimental group treated with / kg was observed that the production of 1L-4 and IL-13 was significantly inhibited.

Inhibition of Cytokine Production by Verproside in Bronchoalveolar Fluid  Kinds  cytokine (pg / ml)  IL-4  IL-13  (-) Control  0.1 ± 0.5  0.1 ± 1.0  OVA treatment group 14.1 ± 6.1 ^* 178.5 ± 96.4 ^*  OVA + Verphoxide (30mg / kg) treated group (% inhibition) 5.0 ± 3.9 ^** (64.5%) 44.8 ± 27.7 ^** (74.9%)  OVA + montelukast (30 mg / kg) treatment group inhibition rate (%) 4.3 ± 3.1 ^** (69.5%) 27.6 ± 14.7 ^** (84.5%)  * significant difference from normal control (NC) group p <0.005 ** significant difference from OVA treated group, p <0.05

Experimental Example  3. Bronchial alveolar fluid  Eosinophil analysis

100 μl of the bronchial alveolar fluid obtained in Experimental Example 2-1 was placed on a slide and centrifuged using a cytospin device (Hanil, Korea) to fix cells on the slide. Total cell number excluding dead cells stained with trypan blue was calculated on a hematocytometer and measured in three replicates (Daigle I. et al., Swiss Med. Wkly., 131 , pp 231-237, 2001). Eosinophil counts were determined after staining with Diff-Quick reagent (Sysmex, Cat No. 38721, Switzerland) and calculated in the same manner as total cell numbers. Statistical analysis was performed by Student's t-test, and significance determination was expressed as p value.

As a result of the experiment, in the OVA treated group sensitized with egg white albumin (OVA) and challenged with PBS as shown in Table 4, the total number of inflammatory cells increased more rapidly than the negative control group without negative airway. The number of eosinophils was 75.4%. In this study, the influx of inflammatory cells was significantly decreased in the group treated with Verproside 30 mg / kg with egg white albumin. Inhibition of bronchial alveolar fluid inflow of total inflammatory cells and eosinophils was 79.3 ± 13.2% and 86.3 ± 7.3%, respectively. Appeared. In the group treated with montelukast as a control drug, total inflammatory cells and eosinophils showed 78.3 ± 12.0% and 80.5 ± 11.1%, respectively.

 Inhibition of Inflammatory Cell Influx of Bronchoalveolar Liquor by Verproside  Total inflammatory cells  Eosinophils Cell count (× 10 ⁴ cells / mouse)  Inhibition (%) Cell count (× 10 ⁴ cells / mouse)  Inhibition (%)  (-) Control  2.27 ± 0.55  -  0.11 ± 0.04  -  OVA treatment group 40.45 ± 16.36 ^*  - 30.48 ± 12.45 ^*  -  OVA + Verproside (30mg / kg) 8.36 ± 5.32 ^**  79.3 4.18 ± 2.21 ^**  86.3 OVA + montelukast (30mg / kg) 8.78 ± 4.87 ^**  78.3 5.94 ± 3.39 ^**  80.5  * significant difference from normal control (NC) group p <0.001 ** significant difference from OVA treated group, p <0.005

Experimental Example 4 Tissue Analysis

Immediately after obtaining bronchoalveolar lavage fluid in each experimental group of Experimental Example 2, the lung tissue was immersed in a 10% medium formalin solution for 24 hours, and then the tissues were placed in paraffin to use a microtome (microtome, SLEE MAINZ, Germany) with a thickness of 6 μm. Sections were prepared, and the sections were stained with hematoxylin (mayer's hematoxylin solution, Sigma, MHS-16) and eosin (Eosin Y solution ascoholic, Sigma, HT110-1-32) and analyzed for tissue. Sections were stained with PAS (periodic acid Schiff) to analyze the secretion of mucus.

As a result of the above experiment, as shown in FIG. 2, in the histological examination of the bronchial tissue, the thickness and the number of inflammatory cells of the tracheobronchial tissue were sensitized with egg white albumin and the secondary antigen-administered rats were administered to the airway and the normal airway without normal airway sensitization. Significantly more inflammatory cells were introduced around the blood vessels. In the case of the treatment with beproside at 30 mg / kg, the influx of inflammatory cells around the airway was significantly reduced and the thickness of bronchial tissue was reduced. Similar to treatment. 0-5 (0; none, 1; few inflammatory cells inflow, 2; 1 layer of inflammatory cells inflow around the airway, 3; 2-4 layers around the airways) Inflammatory cell influx, 4; inflammatory cell influx around the airways, 5; inflammatory cell influx in several layers around the airways), as shown in Table 5 below. Inhibitory effect on inflammatory cell influx.

Inhibitory Effect of Verprosid on Bronchial Tissues Inflammation Index (-) Control 0.52 ± 0.23 OVA treatment group 3.01 ± 0.61 ^* OVA + Verproside (30mg / kg) 1.89 ± 0.45 ^** OVA + montelukast (30mg / kg) 1.72 ± 0.34 ^** * significant difference from normal control (NC) group p <0.005 ** significant difference from OVA treated group, p <0.05

On the other hand, as a result of staining the mucous secretion of airway tissue by PAS staining method, as shown in Figure 3, mice sensitized with egg white albumin and administered secondary antigens secreted a large amount of mucus from goblet cells of airway tissue. (Purple region) was observed, and when the verproside was treated with 30 mg / kg, it was confirmed that the mucus secretion was significantly reduced. The amount of mucous secretion was calculated by area and analyzed with a score of 0-4 (0; none, 1; 25% or less, 2; 25-50% 3; 50-75%, 4; 75% or more). As shown in 6, the beproside treatment group showed a significant mucus secretion inhibitory effect.

Inhibitory Effect of Verprosid on Mucus Secretion in Bronchial Tissues Inflammation Index (-) Control 0 OVA treatment group 3.60 ± 0.34 OVA + Verproside (30mg / kg) 1.43 ± 0.20 ^** OVA + montelukast (30mg / kg) 1.53 ± 0.23 ^* * significant difference from normal control (NC) group p <0.005 ** significant difference from OVA treated group, p <0.05

Experimental Example  5. Measurement of acute inflammation inhibitory activity

Six ICR male mice were used as a group, and carrageenan 1% physiological saline solution was injected subcutaneously into the center of the sole of the foot. The drug was suspended in 500 ul of phosphate buffer solution 1 hour before administration of the baseline, and orally administered with a dose of 20 mg / kg, and 100 mg / kg of aspirin was used as a control. As a result of the experiment, as shown in Table 7, the edema increased after 4 hours and decreased at 5 hours. Significantly increased, inhibition rate was similar to that of high aspirin.

 Untreated group edema rate (%) Verproside treated group (20 mg / kg) Aspirin treated group (100 mg / kg)  Foot edema rate (%)  Inhibition Rate (%)  Foot edema rate (%)  Inhibition Rate (%)  0 hr  100  -  -  -  -  One 119.2 ± 15.6 139.1 ± 16.4  -16.6 ± 13.8 139.3 ± 40.6 -16.8 ± 34.0  2 158.0 ± 9.0 147.9 ± 9.5  6.4 ± 6.0 170.4 ± 34.5 -7.8 ± 21.8  3 194.9 ± 12.4  177.6 ± 12.3  8.9 ± 6.3 180.6 ± 28.6 7.3 ± 14.7  4 205.9 ± 19.0  193.2 ± 4.3  6.1 ± 2.1 196.9 ± 15.4 4.3 ± 7.5  5 201.6 ± 5.1  206.3 ± 7.2  -2.3 ± 3.6 198.5 ± 12.6 1.5 ± 6.3

Examples of the pharmaceutical compositions containing the compounds of the present invention will be described, but the present invention is not intended to be limited thereto, but is intended to be described in detail.

Formulation Example 1 Preparation of Powder

Verproside 300 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation example  2. Preparation of Tablets

Verproside 50 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

Verproside 50 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

Verproside 50 mg

Appropriate sterile distilled water for injection

pH adjuster

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

Verproside 100 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method of preparing a liquid solution, each component is added and dissolved in purified water, lemon flavor is added to the mixture, and then the above ingredients are mixed, purified water is added to adjust the total amount to 100 ml, and then filled in a brown bottle. The solution is prepared by sterilization.

Formulation Example 6 Preparation of Healthy Food

Verproside 1000 mg

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

vitamin 0.13 mg

Vitamin B₂ 0.15 mg

Vitamin B₆ 0.5 mg

Vitamin B₁₂ 0.2 μg

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

Folate 50 ㎍

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation example  7. Manufacture of health drinks

Verproside 1000 mg

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

After mixing the above components in accordance with a conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

Although the composition ratio is mixed with a component suitable for a favorite beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

As described above, the catalpol derivative compounds of the present invention are immunoglobulin E (IgE) and interleukin-4 (IL-4), interleukin-13 (IL-13) of bronchial alveolar fluid in an ovalbumin induced asthma model. Pharmaceutical composition for the prevention and treatment of inflammatory diseases, allergies and asthma containing catalpol derivatives as an active ingredient by inhibiting the production of c), inhibiting eosinophil growth, and inhibiting edema in carrageenan-induced animal models. Or as a dietary supplement.

Claims (4)

A pharmaceutical composition for the prevention or treatment of inflammation, allergies and asthma diseases comprising the catalpol derivative of formula (I) or a pharmaceutically acceptable salt thereof:

                         (Ⅰ) In the above formula R is a benzoyl group or cinnamoyl group each independently substituted with a hydrogen atom or at least one hydroxy C ₁ -C ₃ lower alkyl group or a lower alkoxy group. According to claim 1, wherein the R substituent is a hydrogen atom, 3,4-dihydroxybenzoyl (3,4-dihydroxybenzoyl), 4-hydroxy-3-methoxybenzoyl (4-hydroxy-3-methozybenzoyl), 3 -Hydroxy-4-methoxybenzoyl3-hydroxy-4-methozybenzoyl), 4-hydroxybenzoyl, 3,4-dimethoxybenzoyl (3,4-dimethoxybenzoyl) 3,4-dihydroxy A pharmaceutical composition comprising a compound that is cinnamoyl (3,4-dihydroxycinnamoyl) or 3-hydroxy-4-methoxycinnamoyl (3-hydroxy-4-methoxycinnamoyl). A dietary supplement comprising the catalpol derivative of claim 1 having anti-inflammatory, anti-allergic and anti-asthmatic activity and a food supplement acceptable food additive. The dietary supplement according to claim 3, which is a tablet, capsule, pill or liquid.

Images