KR20090129561A - Composition comprising the extract of coptidis rhizoma for preventing and treating respiratory organ disease - Google Patents

Abstract

PURPOSE: A composition containing Coptidis Rhizoma extract for preventing and treating respiratory disease is provided to ensure high anti-histamine activity and discharge of phlegm. CONSTITUTION: A composition for discharge of phlegm contains Coptidis Rhizoma extract as an active ingredient. The Coptidis Rhizoma extract is crude extract or polar solvent soluble extract of crude extract. The crude extract is extract using water, straight or branched alcohol of 1-6 carbon atoms or their mixture solvent. The Coptidis Rhizoma extract contains berberine, palmatine, coptisine, and jatrorhizine. The composition is used in the form of powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, suppository or sterilization injection solution.

Description

Composition comprising the extract of Coptidis Rhizoma for preventing and treating respiratory organ disease}

The present invention relates to the expectorant activity, antitussive activity and antihistamine activity effect of the extract of the yellow lotus, provides a composition useful for the prevention and treatment of antitussive, expectorant or respiratory diseases containing the extract as an active ingredient.

The respiratory system of our body is one of the organs that constantly defends against physical and chemical stimuli. When the stimulant enters the respiratory tract, most of the stimulant gets caught in the nose and does not enter the respiratory tract, but the fine stimulant passed through it is surrounded by mucus, The tiny cilia located on the airway wall send them out or up, which is the basic principle of coughing. Cough, as mentioned earlier, is a reflexive defense mechanism caused by airway mucosal stimulation, and when a persistent cough is caused by excessive stimulation, it reduces and worsens the quality of life of the patient. In the same principle as coughing, when dust or irritants are introduced from the outside, the body's bronchus moves muscles with saliva and pushes them to the outside. This is the cause of sputum formation, which causes dark purulent sputum due to inflammation of the bronchus and lungs. Will be.

As a result, cough and sputum are caused by physical and chemical factors such as cold air, external foreign substances, including pathogenic microorganisms, air pollutants, and allergens, which can be divided into three categories.

First, stimulation of cough receptors in the larynx, trachea, bronchus, pharynx, sinuses, diaphragm due to physical and chemical factors, the stimulus is transmitted to the cough center of the soft zone of the brain and cough reflex occurs. The mechanisms of antitussives, which are closely related to these irritant cough reflexes, are classified into central and peripheral, and the mechanisms of central antitussives are drugs that control cough by inhibiting the relieving center of soft water, such as codeine and dextromethorphan ( dextromethorphan, noscapine, ephedrine, zipeprol, etc. These components often cause side effects such as cardiovascular thrombosis, myocardial infarction, respiratory depression, sedation, and gastrointestinal disorders. There is. Peripheral antitussives act on the kidney's kidney receptor (stretch receptor) to control cough, which is relatively safe to compensate for the disadvantages of central antitussives, and the development of such antitussives has recently been tried a number of representative drugs Levopropizine, benzonatate, benzozaine, benzyl alcohol, and the like are used.

Second, when the parasympathetic nervous system is activated due to physico-chemical factors, bronchial smooth muscle contracts and thus symptoms such as bronchial spasms and bronchial contractions may occur. At this time, the beta 2 agonist stimulates the beta 2 receptor to relax the smooth muscle, and the xanthine derivative increases cAMP or inhibits bronchial smooth muscle by inhibition of phosphodiesterase. It acts as a direct relaxation, increasing mucus secretion while dissolving mucus and promoting ciliary movement. Drugs exhibiting this action include beta 2 agonists such as clenbuterol, bambuterol and formoterol, and xanthine derivatives such as theophylline and aminophylline. aminophylline), acepiphylline (acepifylline), bamifylline and the like.

In addition, there is a mucus solubilizer or expectorant that dissolves the produced mucus to reduce viscosity and promote bronchial cilia, thereby promoting sputum discharge. Airway mucus has several important functions in prayer. That is, it protects the airway from particles entering the airway from the outside, maintains the humidity of the intake air, and serves to remove the inhaled chemicals and gases by binding to the mucus protein, cilia. Mucus also has substances that play an important role in immune function, such as immunoglobulin A (IgA), lysozyme, lactoferrin, and the like. Most expectorants have been used as folk medicines or experience prescriptions for thousands of years using plants, and the compounds include cysteine derivatives such as carbocysteine, N-acetylcysteine, and cysteine ethyl ester hydrochloride. Is being used. The above-mentioned cysteine derivatives are not completely satisfactory when used as an expectorant, and in particular, N-acetyl cysteine and cysteine ethyl ester hydrochloride have both high toxicity and chemical instability. Therefore, the development of expectorants with good physical and chemical stability and less side effects, toxicity and good expectorant effects has been required.

Recently, research on antitussive expectorants derived from natural products has been actively conducted. Representative studies related to this include stimulating the vagus nerve of the airway to promote secretion of the mucosa, expressing expectoration and at the same time, n-supplementary role of antitussive action. There are also reports of plants containing saponins, alkaloids, etc. (Sook Young Kim et al . , Kor . J. Pharmacogn . 19 (2): 133-140, 1998).

Third, inflammatory mediators are released from mast cells due to physical and chemical factors. Mast cells contain small cytoplasmic granules containing inflammatory mediators such as histamine and cytokines. When exposed to an allergen, the body produces an antibody called IgE in B cells, which attaches to the surface of mast cells. Subsequent exposure to allergens causes the release of allergens, including histamine, a potent inflammatory mediator, from IgE-attached mast cells, resulting in increased secretions in the airways, effusions from inflammation, inhaled foreign bodies, and bacteria. Leads to. Existing drugs such as codeine, widely used as antitussives, have excellent sedative effects on breathing and coughing, but have side effects that release histamine by causing degranulation of mast cells.

In addition, based on the above description, the mechanism and function closely related to antitussive expectoration, antibacterial action to enable the removal of the causative agent and prevention of secondary infection, anti-inflammatory action to inhibit the synthesis of inflammatory mediators, histamine from mast cells Antihistamine activity that inhibits secretion, antioxidant activity that prevents inflammation and tissue damage, elastase activity release that releases inflammatory mediators from immune cells, hyaluronidase activity involved in vascular permeable inflammatory reactions Inhibition, etc. will be a function that requires.

Thus, the present inventors have completed the present invention by confirming that the extract of the antitussive or expectorant is excellent in the antitussive, expectorant and antihistamine activity effects.

In order to carry out the above object, the present invention provides a composition for antitussive or expectorant containing sulfur extract as an active ingredient.

Herbal medicine (Coptidis Rhizoma), a herb used in the present invention, is a perennial herb belonging to Ranunculus family. The rhizome is called Hunan, yellow, branched, and beard roots are dense. It has been known to have pharmacological effects such as antimicrobial action, blood pressure lowering action and anti-inflammatory action. In oriental medicine, it is used as a gastric agent, central nervous system agent, uterus, bladder, bronchial and nervous system agents. It is known to contain alkaloids such as berebrine, palmatine, coptisine, berberastine, magnoflorine, etc. , Dohae Hyangdae (Medicinal Herbs), 490-493, 1990).

However, the effects of these extracts and components on antitussive, expectorant and antihistamine activities have not been studied.

'Extract' as defined herein is characterized in that the crude extract of the yellow lotus or a polar solvent soluble extract of the crude extract.

The crude extract as defined herein is a solvent selected from water, including purified water, straight-chain or branched alcohols having 1 to 6 carbon atoms or mixed solvents thereof, preferably water, 10-70% (v / v) aqueous ethanol solution or water. It is characterized by a saturated butyl alcohol.

The polar solvent soluble extract as defined herein is characterized in that the linear or branched alcohol having 1 to 6 carbon atoms, preferably saturated butyl alcohol, and the fraction is the methylene chloride and methanol (30: 1 to 7). : 1 (v / v)) is a mixed solvent, characterized in that the fraction obtained by performing silica gel column chromatography .

The rhubarb extract or fraction of the present invention is characterized in that it contains berberine, palmin, copticin, columamine and jathrorizine, preferably berberine (0.5 to 47.0 parts by weight): palmin (0.2 to 21.4 parts by weight) ): Copticin (0.1 to 18.0 parts by weight): columamine (0.1 to 3.2 parts by weight): jatrorizine (0.1 to 2.6 parts by weight) characterized in that it comprises within the range.

Hereinafter, the method for obtaining the crude extract, the polar solvent soluble extract or fraction of the crude extract of the present invention will be described in detail.

The crude extract of the yellow rye of the present invention is chopped the dried yellow rye, about 3 to 20 times the weight (kg), preferably about 5 to 15 times the water, C ₁ to C ₆ linear or branched alcohol Or a mixed solvent thereof, preferably water, 10-70% (v / v) aqueous ethanol solution or saturated butyl alcohol, at an extraction temperature of 40 ° C to 110 ° C, preferably 55 ° C to 90 ° C. The crude extract of the present invention may be obtained by filtration, concentration under reduced pressure, or drying the extract obtained by using cold soaking, hot water extraction, ultrasonic extraction, reflux cooling extraction for 20 hours, preferably about 1 to 10 hours.

The crude extract was filtered to collect the filtrate, and the residue was added with 4-7 times the weight of the crude drug raw material water, an aqueous alcohol solution or a saturated butyl alcohol, and warmed again to extract for 2-5 hours, filtered and mixed with the previous filtrate. Increase the extraction efficiency If the amount of solvent is too small, the stirring becomes difficult and the solubility of the extract is low, the extraction efficiency is lowered. If the amount is too large, the amount of lower alcohol used in the next purification step is increased, which is not economical and may cause handling problems. . In the present invention, the method of re-extracting after the first extraction is adopted. Even in the case of mass production of the herbal extracts, even though the filtration is effective, the loss occurs because the water content of the herbal medicine itself is high, so the extraction efficiency is reduced only by the first extraction. This is to prevent this. In addition, as a result of verifying the extraction efficiency of each step, it was found that about 80 to 90% of the total extraction amount is extracted by the second extraction, and the third or more multi-stage extraction is not economical.

In addition, the polar solvent soluble extract of the present invention is suspended in the crude extract of about 2 to 10 times, preferably about 3 to 7 times the volume of water, the polar solvent such as saturated butyl alcohol 0.5 to 3 times of the suspension, It is preferably prepared by adding the same amount to 2 times and extracting the polar solvent soluble layer for 1 to 5 times, preferably 2 to 3 times, followed by concentration under reduced pressure.

The lower alcohol fraction obtained after layer separation is concentrated under reduced pressure at 50-60 ° C. to remove the solvent remaining in the sample. The concentrate thus obtained is azeotropically concentrated two to three times with water 25 to 50 times the total amount of the concentrate, and suspended again in an equal amount of water. The reason for azeotropic concentration with water at the time of the concentrated drying is to effectively control the content of the lower alcohol remaining in order to use the obtained herbal extract as a raw material for pharmaceuticals.

As described above, the extract obtained by extraction with water, C ₁ to C ₆ straight chain or branched alcohol, or a mixed solvent thereof is filtered and concentrated, and then unnecessary proteins, polysaccharides and fatty acids contained in the filtrate. In the present invention, impurities are purified by performing a layer separation two to four times with the filtrate and the same amount of lower alcohol to obtain a solvent fraction. In this case, as the lower alcohol, C1-C6 alcohol, preferably butyl alcohol, propyl alcohol or isopropyl alcohol, and when the amount of lower alcohol is less than that of the filtrate, fine particles are formed by unnecessary components such as fatty acids. Therefore, the separation of the layers is not smooth and the extraction content of the active ingredient is lowered, which is not efficient.

In addition, the sulfur extract obtained in the above was mixed with methylene chloride and methanol (30: 1-7: 1 (v / v)), and subjected to silica gel column chromatography, divided into five subfractions to obtain physiology for each fraction. By conducting an activity experiment, active fractions and components were identified.

As a result of analyzing the components of the extract and fractions of the present invention, it was confirmed that berberine, palmatin, copticin, columamine, and jathrorizine are included.

Accordingly, another aspect of the present invention is directed to a composition for the prevention or treatment of antitussive, expectorant or respiratory diseases containing at least one selected from the group consisting of berberine, palminth, copticin, columamine, and jathrorizine as an active ingredient. It is about. Preferably, the composition may contain 0.5 to 47.0 parts by weight of berberine, 0.2 to 21.4 parts by weight of palmatin, 0.1 to 18.0 parts by weight of copticin, 0.1 to 3.2 parts by weight of columamine, and 0.1 to 2.6 parts by weight of jathrorizine. have.

It will be described in detail how to analyze the components contained in the extract.

The crude lotus extract and the small fraction obtained by the above method were subjected to a solvent gradient of 0.2% phosphoric acid solution in a mobile phase: methanol (9: 1 to 6: 4 → 6: 4 to 5: 5 → 5: 5 to 0:10). It can be carried out to obtain the active ingredients, columamine and jatrorizine.

In addition, the standard berberine, palmin, and copticin are commercially available.

The present invention provides a pharmaceutical composition for the prevention and treatment of respiratory diseases, containing the extract of the yellow lotus obtained by the production method as an active ingredient.

Respiratory disease as defined herein includes emphysema, chronic bronchitis, bronchial asthma, bronchial adenoma, isolated pulmonary nodules, pulmonary tuberculosis, empyema, lung abscess, cold, flu or histiocytosis of the lungs, preferably emphysema, chronic with cough or sputum Bronchitis, bronchial asthma, pneumonia, cold, flu.

The composition for preventing and treating respiratory diseases of the present invention comprises 0.001 to 99% by weight, preferably 0.1 to 50% by weight of the compound, based on the total weight of the composition.

However, the composition as described above is not necessarily limited thereto, and may vary according to the condition of the patient and the type and extent of the disease.

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

Compositions comprising extracts according to the invention are formulated 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. Carriers, excipients and diluents which may be used in combination with the extract, and which may be included in the composition comprising the extract 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 the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Oral liquid preparations include suspending agents, liquid solutions, 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 extracts of the present invention vary depending 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, and may be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is 1 mg / kg to 1000 mg / kg, preferably 50 mg / kg to 500 mg / kg, more preferably 150 mg / kg to 300 mg / kg per day It is good to administer. Administration may be administered once a day or may be divided several times. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

The composition of the present invention can be administered to mammals such as rats, 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 addition, the present invention provides a health functional food for the prevention and improvement of antitussive, expectorant or respiratory diseases containing the extract of the yellow lotus as an active ingredient.

The composition containing the extract of the present invention can be used in various ways, such as drugs, foods and drinks for the prevention and improvement of respiratory diseases. Foods to which the extract of the present invention may be added include, for example, various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages. have.

Since the extract itself of the present invention has little toxicity and side effects, it is a drug that can be used safely even when taken for long periods of time.

The extract of the present invention may be added to food or beverage for the purpose of preventing and improving respiratory diseases. At this time, the amount of the extract in the food or beverage is generally added to the health food composition of the present invention to 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.02 to 10 g based on 100 ml, preferably Can be added in a ratio of 0.3 to 1 g.

In addition to containing the extract as an essential ingredient in the indicated proportions, the health beverage composition of the present invention has no particular limitation on the liquid component, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates are monosaccharides such as disaccharides such as glucose, fructose and the like, such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin and the like. Sugars 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 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 composition of the present invention includes 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 beverages, and the like. The compositions of the present invention may also contain pulp 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 generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

Huang lotus extract of the present invention can be used as a pharmaceutical composition and health functional food useful for the prevention and treatment of respiratory diseases by confirming the expectorant activity, antitussive activity and antihistamine activity.

Hereinafter, the present invention will be described in detail by Examples and Experimental Examples.

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

Example  One. goldthread  Preparation of Extracts (A)

1-1. goldthread Crude extract  ( CR - GA Manufacturing

After removing the condensate by washing with the water rhubarb purchased from Gyeongdong Market, 250 g of dried kumquat was extracted with 1.5 L of water, twice with hot water at 80 ° C. for 3 hours, and the extract was filtered and concentrated under reduced pressure to give 62.5 g of crude extract ( 25% yield compared to the original drug), which was hereinafter referred to as 'CR-GA' and was used in Example 1-2 below.

1-2. goldthread  Polar Solvent Soluble Extract ( CR - GA -1) Preparation

Example 1-1 was suspended in 0.5 L of water applied to the CR-GA 62.5 g obtained from, and then can here saturated butyl alcohol was added to 1 L, separated layer twice, saturated butyl alcohol fraction only be collected dried Concentrated under reduced pressure until. 0.4 L of water and most of the butyl alcohol and water was evaporated, azeotropically concentrated, and then repeated twice. Finally, 0.1 L of distilled water was added and suspended, followed by freeze-drying. (7.92% yield compared to the original herbal medicine), which was hereinafter referred to as 'CR-GA-1' and was used as a sample of the following experimental example.

Example  2. goldthread  Preparation of Extract (B)

2-1. goldthread Crude extract  ( CR - GB Manufacturing

After removing the condensate by washing with water, the yellow lotus purchased from Gyeongdong Market was extracted with 2.0 g of 50% (v / v) ethanol solution and refluxed twice at 80 ° C. twice for 3 hours. After filtration and concentration under reduced pressure to obtain a crude extract of 57.5g (23% yield compared to the crude drug), which was named below 'CR-GB', was used in Example 2-2 below.

2-2. goldthread  Polar Solvent Soluble Extract ( CR - GB -1) Preparation

0.3 L of water was added to 57.5 g of CR-GB obtained in Example 2-1 and suspended. 0.6 L of saturated butyl alcohol was added thereto, followed by two layer separations, and then only the butyl alcohol fraction was collected and dried. Concentrated under reduced pressure. Most of the butyl alcohol and water were evaporated, 0.2 L of water was added, the azeotropic concentration was repeated, and then repeated twice. Finally, 0.2 L of distilled water was added and suspended, followed by freeze-drying. (12.7% yield compared to the original herbal medicine), which was hereinafter referred to as 'CR-GB-1' and was used as a sample of the following experimental example.

Example  3. goldthread  Preparation of Extract (C)

3-1. goldthread  Extract ( CR - GC Manufacturing

After removing the condensate by washing with water, the crude rhubarb purchased from Gyeongdong Market was extracted by 2.5 g of saturated n-butyl alcohol, reflux-cooled twice at 85 ° C. twice for 3 hours. Thereafter, the extract was concentrated under reduced pressure after filtration, and then azeotropically concentrated by adding 0.2 L of water in a state where most of the butyl alcohol was evaporated, and then repeated twice, and finally suspended by adding 0.2 L of distilled water. It was dried to obtain 32.8 g (13.1% of the original drug) in the form of a powder, which was hereinafter referred to as 'CR-GC', was used in the following experimental example.

Example  4. goldthread  Of extract Small fraction  Produce

20 g of CR-GB-1 of Example 2 was mixed with methylene chloride and methanol (30: 1 to 7: 1 (v / v)) by silica gel column chromatography, and divided into five fractions. The fraction is Fr. 1 is 0.8 g (hereinafter referred to as 'Fr. 1'), Fr. 2 is 2.6 g (hereinafter referred to as 'Fr. 2'), Fr. 3 is 5.5 g (hereinafter referred to as 'Fr. 3'), Fr. 4 is 2.8 g (hereinafter referred to as 'Fr. 4'), Fr. 5 was 6.5 g (hereinafter referred to as 'Fr. 5') and was used as a sample of the following experimental example.

Experimental Example  One. goldthread  Determination of expectorant activity of extract

In order to measure the expectorant activity of the rhubarb extract of Examples 1 to 3, using Engler et al. (Engler H, Szelenyi I, J. Pharmacol. Moth. 11, 151-157, 1984 ,, Bao -quin Lin et., Pulmonary Pharmacology & Therapeutics 21, 259-263, 2008.), the experiment was carried out in the following process.

 30 minutes after oral administration of the positive control drug (ambroxol, Sigma) and the test drug (Rhubarb extracts of Examples 1 to 3), phenol red was intraperitoneally injected (500 mg / kg dose, Dissolve in saline solution). After 30 minutes, anesthetized with diethyl ether, the abdominal aorta was cut and bled, and then the entire trachea was excised. The separated organs were placed in 1 ml of physiological saline, washed for 30 minutes, centrifuged at 10,000 rpm for 5 minutes at room temperature, and 1N caustic soda (NaOH) was added to the supernatant (0.1 ml of 1N NaOH per 1 ml of supernatant). After the absorbance at 546 nm was measured the expectorant activity as a concentration of phenol red, the results are shown in Table 1 below.

division Dosage (mg / kg) Sputum discharge capacity (%) Example 1 CR-GA 500 29 CR-GA-1 500 36 Example 2 CR-GB 500 32 CR-GB-1 500 41 Example 3 CR-GC 500 38 Positive control group (Ambroxol) 250 32

As a result, as shown in Table 1, it was found that the overall activity is excellent, in particular, it was confirmed that the sputum discharge capacity of the CR-GB-1 of Example 2 shows the most excellent activity.

In addition, the activity was measured by the dosage amount (50, 100, 150, 200, 500 mg / kg) of CR-GB-1 of Example 2, the results are shown in Table 2 below.

division Dosage (mg / kg) Sputum discharge capacity (%) Example 2 CR-GB-1 50 4 100 22 150 32 200 33 500 41 Positive control group (Ambroxol) 250 30

As a result, as shown in Table 2, when the dose of CR-GB-1 of Example 2 is 500 mg / kg, it was confirmed that the sputum discharge capacity showed the best activity.

Experimental Example  2. goldthread  Antitussive Activity of Extracts

In order to measure the antitussive activity of the sulfur extracts of Examples 1 to 3, using Tanaka et al. (Motomu Tanaka and Kei Maruyama., J. Pharmacol. Sci. 93, 465-470, 2003., Daoui , Cognon, Naline et., Am. J. Respir. Crit. Care. Med. 158, 42-48, 1998.), experiments were carried out in the following process.

After 1 hour of oral administration of a test drug to male guinea pigs (6 weeks old, Samtaco Bio Korea), guinea pigs were placed in a plastismograph chamber (Buxco, USA), and then sprayed with cough inducing citric acid (Sigma). Causing a cough. As a positive control, theobromine (Sigma), which is used as an antitussive agent, was used. The guinea pig was exposed to 2 M citric acid for 10 minutes, and the cough water generated at this time was measured for 15 minutes, and the results are shown in Table 3 below.

division Dosage (mg / kg) Cough suppression ability (%) Example 1 CR-GA 200 33 CR-GA-1 200 46 Example 2 CR-GB 200 35 CR-GB-1 200 52 Example 3 CR-GC 200 44 Positive control group (Theobromine) 50 57

As a result, as shown in Table 3, it was found that the overall activity is excellent, in particular, it was confirmed that the sputum discharge capacity of the CR-GB-1 of Example 2 shows the most excellent activity.

In addition, the activity was measured by the dosage amount (50, 100, 150, 200 mg / kg) of CR-GB-1 of Example 2, the results are shown in Table 4 below.

division Dosage (mg / kg) Cough suppression ability (%) Example 2 CR-GB-1 50 7 100 25 150 37 200 52 Positive control group (Theobromine) 50 57

Experimental results, as shown in Table 4, when the dosage of CR-GB-1 of Example 2 is 200 mg / kg, cough It was confirmed that the inhibitory effect shows the best activity.

Experimental Example  3. goldthread  Determination of Antihistamine Activity of Extracts

In order to measure the antihistamine activity of the rhubarb extract of Examples 1 to 3, using a method such as Honuchi (Masako Honuchi and Yoshiyuki Seyama, J. Health Sci., 52 (6), 711 ~ 717, 2006. , Naoki Inagaki et. Biol. Pharm. Bull. 24 (7), 829-834, 2001.), the experiment was carried out by the following process.

Antihistamine effects were measured using celiac cells of male rats (7 weeks old, Samtaco Bio Korea) sensitized with egg white albumin. Ketotyfen (Ketotifen, Sigma) was used as a positive control. Rats sensitized with egg white albumin were orally administered with the test drug and orally with the drug for three days, and then the peritoneal mast cells were isolated. The isolated celiac mast cells (2 × 10 ⁵ cells / ml) were treated with the drug again. Finally, compound 48/80 (compound 48/80, Sigma), which is a substance that induces degranulation of mast cells to secrete histamine by nonimmunological method, is treated in the same concentration at a concentration of 10 μg / ml. The amount of free histamine was quantified to determine whether the test substances inhibited histamine release from mast cells. Relative amounts relative to the amount of histamine released from mast cells in rats that were not administered any drug were measured, and the results are shown in Table 5 below.

division Oral Dose (mg / kg) Mast Cell Concentration (mg / ml) Histamine Release Rate (%) Example 1 CR-GA 200 0 41 0.1 38 1.0 33 10.0 26 CR-GA-1 200 0 44 0.1 35 1.0 29 10.0 23 Example 2 CR-GB 200 0 42 0.1 38 1.0 31 10.0 27 CR-GB-1 200 0 39 0.1 34 1.0 27 10.0 21 Example 3 CR-GC 200 0 41 0.1 37 1.0 30 10.0 22 Positive control group (Ketotifen) 5 0 43 0.01 41 0.1 32 1.0 18

As a result, as shown in Table 5, it was found that the overall activity is excellent, in particular, it was confirmed that the anti-histamine effect of CR-GB-1 of Example 2 shows the most excellent activity.

In addition, the activity was measured by the dosage amount (100, 200, 400 mg / kg) of CR-GB-1 of Example 2, the results are shown in Table 6 below.

division Oral Dose (mg / kg) Mast Cell Concentration (mg / ml) Histamine Release Rate (%) Example 2 CR-GB-1 100 0 49 0.1 40 1.0 36 10.0 30 200 0 42 0.1 34 1.0 27 10.0 21 400 0 31 0.1 26 1.0 23 10.0 18 Positive control group (Ketotifen) 5 0 43 0.01 41 0.1 32 1.0 18

As a result, as shown in Table 6, when the dose of CR-GB-1 of Example 2 is 400 mg / kg, it was confirmed that the antihistamine effect showed the best activity.

Experimental Example  4. Active Small fraction  Jinhae activity measurement

In order to measure the antitussive activity of the five small fractions prepared in Example 4, the experiment of the antitussive activity was performed in the same manner as in Experimental Example 2, and the experimental results are shown in Table 7 below.

division Dosage (mg / kg) Cough suppression ability (%) Fr. One 60 41 Fr. 2 60 32 Fr. 3 60 62 Fr. 4 60 34 Fr. 5 60 24 Positive control group Theobromine 50 57

As a result, as shown in Table 7, it was found that the overall activity is excellent, in particular, it was confirmed that the antitussive activity of Fr.3 is excellent.

Experimental Example  5. Active Small fraction  Determination of antihistamine activity

In order to measure the antitussive activity of the five small fractions prepared in Example 4, the same experiment as in Experimental Example 3 was performed, and the experimental results are shown in Table 8 below.

division Oral Dose (mg / kg) Mast Cell Concentration (mg / ml) Histamine Release Rate (%) Fr. One 60 0 55 0.1 50 1.0 42 10.0 39 Fr. 2 60 0 59 0.1 54 1.0 48 10.0 41 Fr. 3 60 0 38 0.1 31 1.0 26 10.0 19 Fr. 4 60 0 56 0.1 50 1.0 44 10.0 40 Fr. 5 60 0 66 0.1 62 1.0 55 10.0 51 Positive control Ketotifen 5 0 43 0.01 41 0.1 32 1.0 18

As a result, as shown in Table 8, the overall activity was found to be excellent, in particular, it was confirmed that the antihistamine activity of Fr.3 is excellent.

Experimental Example  6. goldthread  Component Analysis of Extracts

In order to analyze the components for the sulfur extract and fractions of the above examples, experiments were performed using physicochemical analysis such as HPLC, LC / MS, UV-Spectrometer and FT-NMR.

As a result of the experiment, it was confirmed that the extract of the yellow lotus contained berberine, palmin, coptisine, columamine, and jathrorizine, and the component content of the extract of Examples 1 to 4 was analyzed as follows. Is shown in Table 9.

High performance liquid chromatography was measured using a Waters Alliance 2695 model using a Waters PD 2996. The column was YMC Hydrosphere C18 (YMC Hydrosphere C18, S-5㎛, 120nm, 4.6 × 250mm I.D) and the sample temperature was 25 ℃ ± 1, the column temperature was maintained at 30 ℃ ± 1. The sample concentration was prepared in 1mg / ml and injected 10μl and the flow rate was analyzed to 1.0ml / min. In addition, standard materials such as berberine, palminthine, and copticin were commercially available from Sigma, and columamine and jathrorizine were used after separation and purification from the rye. The mobile phase is 0 to 60 minutes (A: B = 9: 1 to 6: 4) and 60 to 70 minutes (A: B = 6) under a gradient of 0.2% phosphoric acid solution (solvent A) and methanol (solvent B). : 4-5: 5), 70 minutes-90 minutes (A: B = 5: 5-0: 10). Calculation of the active ingredient content in the extract indicates the area ratio for each standard in weight percent.

division Content by component (%) Berberine Palm Martin Copticin Columbamine Zatrorizine Example 1 CR-GA 15.6 4.2 4.3 0.6 0.6 CR-GA-1 25.3 4.5 4.7 0.6 0.5 Example 2 CR-GB 22.8 5.6 6.5 1.5 0.9 CR-GB-1 27.0 7.1 5.2 0.9 0.8 Example 3 CR-GC 30.9 7.8 6.2 1.1 1.0 Example 4 Fr. One 0.5 0.2 0.1 0.1 0.1 Fr. 2 10.1 6.7 0.8 0.1 0.1 Fr. 3 47.0 21.4 3.7 3.2 1.5 Fr. 4 5.2 1.4 18.0 0.1 2.6 Fr. 5 0.1 or less 0.1 or less 0.1 or less 0.1 or less 0.1 or less

As a result of the experiment, as shown in Table 9, as a result of comparing the content of the main components by the method of preparation of extracts and fractions, 0.5 to 47.0 parts by weight of berberine, 0.2 to 21.4 parts by weight of palmin, 0.1 to 18.0 parts by weight of copticin, It could be confirmed that it contains 0.1 to 3.2 parts by weight of columamine and 0.1 to 2.6 parts by weight of jathrorizine.

Experimental Example  7. Berberine  Expectorant activity measurement

In order to measure the expectorant activity of the sulfur extract of Example 2, Example 3, and five small fractions prepared in Example 4, an experiment on the expectorant activity was performed in the same manner as in Experimental Example 1, Table 10 below shows the histograms for this activity in FIG. 1 (see FIG. 1).

division Dosage (mg / kg) Sputum discharge capacity (%) Fr. One 125 25 Fr. 2 125 22 Fr. 3 125 44 Fr. 4 125 24 Fr. 5 125 10 CR-GB of Example 2 125 32 CR-GB-1 of Example 2 125 38 CR-GC of Example 3 125 37 Berberine 125 39 Positive control group Ambroxol 250 34

Hereinafter, the preparation examples of the composition including the extract of the present invention, but the present invention is not intended to limit it, but is intended to explain in detail only.

Formulation example  One. Powder  Produce

Rhubarb Extract (Example 1) 20 mg

Lactose 100 mg

Talcuk 10 mg

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

Formulation example  2. Preparation of Tablets

Rhubarb Extract (Example 2) 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

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

Formulation example  3. Manufacture of capsule

Rhubarb Extract (Example 3) 10 mg

Crystalline Cellulose 3 mg

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

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 Injectables

Rhubarb Extract (Example 4) 10 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO _{4 ,} 12H ₂ O 26 mg

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

Formulation example  5. Liquid  Produce

Rhubarb extract (Example 1) 20 mg

Isomerized sugar 10g

5 g of mannitol

Appropriate amount of 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. Manufacture of healthy food

Huang Lotus Extract (Example 2) 1000mg

70 ㎍ Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 ㎍ of vitamin B12

Vitamin C 10 mg

Biotin 10 ㎍

Nicotinamide 1.7 mg

Folic acid 50 ㎍

Calcium pantothenate 0.5 mg

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

Sulfur Extract (Example 3) 1000 mg

Citric acid 1000 mg

Oligosaccharides 100 g

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 a composition that is relatively suitable for the preferred beverage in a preferred embodiment, the compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

Figure 1 shows the effect of sputum emission of the extract and the active fraction and the representative component of the rhubarb through the phenol red emission method using a mouse model.

Claims (14)

 A composition for antitussive or expectorant, containing the extract of rhubarb as an active ingredient. The composition of claim 1, wherein the extract is a crude extract or a polar solvent soluble extract of the crude extract. The composition of claim 2, wherein the crude extract is an extract soluble in a solvent selected from water, linear or branched alcohols having 1 to 6 carbon atoms, or a mixed solvent thereof. The composition of claim 2, wherein the polar solvent soluble extract is an extract soluble in a solvent selected from water, a straight chain or branched alcohol having 1 to 6 carbon atoms, or a mixed solvent thereof. The composition of claim 1, wherein the rhubarb is domestic or imported. The composition of claim 1, wherein the rhubarb extract comprises berberine, palmatin, copticin, columamine, and jathrorizine. An antitussive or expectorant composition containing at least one selected from the group consisting of berberine, palminth, copticin, columamine, and jatririzine as an active ingredient.  The composition of claim 7, wherein the composition comprises 0.5 to 47.0 parts by weight of berberine, 0.2 to 21.4 parts by weight of palmatin, 0.1 to 18.0 parts by weight of copticin, 0.1 to 3.2 parts by weight of columamine, and 0.1 to 2.6 parts by weight of jathrorizine. Characterized by a composition. A composition for the prevention or treatment of respiratory diseases comprising the composition of any one of claims 1 to 8. 10. The composition of claim 9, wherein the respiratory disease is emphysema accompanied by cough or sputum, chronic bronchitis, bronchial asthma, bronchial adenomas, isolated pulmonary nodules, pulmonary tuberculosis, empyema, lung abscess, cold, flu, or histiocytosis of the lung.  10. The composition of claim 9, wherein the composition is formulated as a powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, transdermal, suppository, or sterile injectable solution. 12. The composition of claim 11, wherein the extract comprises 0.001 to 99% by weight of the total weight of the composition.  Health functional food for the prevention or improvement of respiratory diseases comprising the composition of any one of claims 1 to 8. The dietary supplement according to claim 13 in the form of a powder, granule, tablet, capsule or beverage.

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