KR20090129936A - Pharmaceutical composition containing a herbal mixture extract - Google Patents

Abstract

PURPOSE: A therapeutic agent containing ivy leave and Coptidis rhizoma for preventing and treating respiratory disease is provided to suppress bronchoconstriction. CONSTITUTION: A composition for discharging phlegm contains mixture extract of ivy leave and Coptidis rhizome as an active ingredient. The composition is used in the form of powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, suppository, or sterilization injection solution.

Description

Pharmaceutical composition containing a herbal mixture extract

The present invention relates to an agent for the prevention and treatment of respiratory diseases such as antitussive expectoration and asthma, which contains a mixed extract of ivy leaf and coli ( Coptidis rhizoma ) as an active ingredient. It is characterized by exhibiting significantly antitussive action, expectorant action, antihistamine action, bronchial contraction suppression, airway hypersensitivity suppression, bronchial obstruction suppression and inflammatory cell infiltration inhibition than each extract alone. .

The respiratory system of our body is one of the organs that constantly defends against physicochemical stimulation. When the stimulant enters the respiratory tract, most of the stimulant is caught in the nose and cannot enter the respiratory tract. The movement of tiny cilia located on the wall causes them to move up or out, which is the basic principle of coughing.

As described above, the cough is a reflex defense mechanism caused by airway mucosal stimulation, and when a persistent cough caused by excessive stimulation is induced, it reduces and worsens the quality of life of the patient. In addition, when dust or irritants are introduced from the outside in the same principle as coughing, the body's bronchus pushes muscles with saliva and pushes them to the outside, which is the cause of sputum (sputum) formation and inflammation of the bronchus such as the lungs. The result is a thick purulent sputum.

 As a result, cough and sputum are generated by physicochemical factors such as cold air, foreign foreign substances including pathogenic microorganisms, air pollutants, and allergens, which can be classified into three types.

First, stimulation of cough receptors such as larynx, trachea, bronchus, pharynx, sinuses and diaphragm due to physicochemical factors causes stimulation to be transmitted to the cough center of the softened area of the brain and cough reflex occurs. The mechanisms of antitussives, which are closely related to these irritant cough reflexes, are divided into central and peripheral, and the mechanisms of central antitussives are drugs that control cough by suppressing 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 to relieve cough by acting on the kidney receptor (stretch receptor) in the periphery and are relatively safe to compensate for the disadvantages of central antitussives. Recently, the development of such antitussives has been tried a number of representative drugs such as levopropy Levopropizine, benzonatate, benzozaine, benzyl alcohol and the like are used.

Second, when the parasympathetic nervous system is activated due to physicochemical factors, bronchial smooth muscle may contract and thus symptoms such as bronchospasm and bronchial contraction may occur. At this time, the beta 2 agonist (beta2-adrenergic receptor agonist) stimulates the beta 2 receptor to relax the smooth muscle, xanthine derivatives increase cAMP by inhibition of phosphodiesterase or bronchial It plays a role of directly relaxing smooth muscle, increasing mucus secretion, 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 mucolytic agent or expectorant which dissolves the produced mucus to reduce viscosity and promotes cilia movement of bronchus to promote 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.

Third, inflammatory mediators are released from mast cells due to physicochemical 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.

As described above, the existing antitussive expectorant has problems due to side effects, toxicity and chemical instability of the drug itself. In order to solve this problem, recently, research on natural antitussive expectorants has been actively conducted. In this regard, a representative research related to this is stimulating the vagus nerve of the airway, promoting the secretion of mucous membranes to express expectoration and antitussive action. There have been reports of plants containing saponin, alkaloid, etc., which play a secondary role of (Sook Young et al . , Kor . J. Pharmacogn . 19 (2): 133-140, 1998). It is still insignificant.

Asthma, a typical respiratory disease, is a symptom of repetitive seizures such as shortness of breath, coughing, and wheezing breathing (wheezing) .In about 30% of asthma patients, about 80% within one year of life At the age of 4-5 years, the first symptoms of asthma, the incidence of about 10% in Korea, heart asthma and bronchial asthma. The former is a paroxysmal breathing difficulty seen in heart disease or high blood pressure, and is caused by hypersensitivity of the respiratory tract due to pulmonary congestion or tissue congestion and poor prognosis. The latter is a paroxysmal respiratory distress caused by narrowing of breathing due to bronchial spasms, narrowness, and mucosal swelling. In addition, there are cerebral asthma caused by disorders of the respiratory central region, and uremia asthma occurring in patients with nephropathy.

The incidence of asthma varies according to country, race, and age, but according to the 2007 UK report, about 7.9% of adults, 13.7% of children and 9.4% of the elderly are reported in Korea. It is gradually increasing with the increase of stress. Although asthma varies depending on the cause, in modern times when environmental pollution such as pollution is serious, the severity of asthma is maximized, such as the age of onset and the symptoms are prolonged.

The characteristic airway obstruction of asthma occurs by a three-step process. In other words, the contraction of bronchial smooth muscle, thickening of the pulmonary mucosa, and sticky mucus in the bronchus and bronchioles block the airways. Only the contraction of the double bronchial smooth muscle recovers easily.

The exogenous (allergic) asthma attack mechanism is particularly important for antibodies IgE, and some are related to IgG. IgE releases mediators (histamine, SRS-A, ECF-A, NCF, PAF, Kinin, PGs) that activate mast cells to cause hypersensitivity. The mechanism of non-allergy asthma attacks is unknown or mediated by autonomic nerves. Cholinergic stimulation in endogenous patients not only favors mediators such as histamine in mast cells through end organs, but also increases goblet cell secretion, enlarges blood vessels in the lungs, and constricts the trachea, bronchus and giant bronchioles. Bronchospasm and mucus secretion increase.

There is no current cure for asthma, and there are many methods and drugs available to prevent seizures and prevent complications, but they are not satisfactory. The treatment of asthma is primarily for inhalation bronchodilators, oral or injectable bronchodilators (sympathetic and theophylline drugs), stereoforms (for inhalation, oral, injectables, etc.), leukotriene antagonists (montelukast, tranlukast, etc.). , Ziluton, and the like, and anti-allergic drugs (disodium chlorochlorate, chromoline sodium, ketotifen) are used. Airway dilators such as anticholinergic agents and beta-2 receptor agonists have no effect on inflammation that exacerbates the disease and merely relieve symptoms. Steroids, which are known to be effective for inflammation, are problematic for long-term use due to severe side effects. Therefore, a combination of the two is often prescribed, but due to the side effects of steroids developed in the form of inhalants than the oral drug is difficult to take, there is a problem in taking compliance.

Overcoming the limitations of these current drug therapies, there is a need for the development of new therapeutic agents that can fundamentally cure the cause and effectively improve symptoms. However, since respiratory diseases are involved with various leukocyte cells and various cytokines and inflammatory mediators released therein as described above, natural extracts having various ingredients and mechanisms are effective because it is difficult to effectively treat them with a single chemical. Can be

Therefore, the present inventors have studied to develop a therapeutic agent for respiratory diseases such as antitussive expectoration and asthma, the expectorant activity, antitussive activity, antihistamine activity, bronchial contraction inhibitory activity of a mixture of ivy leaf extract and Coptidis rhizoma extract, It was confirmed that airway hypersensitivity inhibitory activity, bronchial occlusion inhibition and inflammatory cell infiltration inhibition activity of the lung tissues were excellent, and these activities were enhanced by the mixing of ivy leaf extract and rhubarb extract than ivy leaf extract itself or rhubarb extract itself. It was confirmed that the present invention was completed.

Accordingly, an object of the present invention is to provide a mixture having an optimal combination ratio with the best pharmacological active synergistic effect by mixing the ivy leaf extract and sulfur extract.

The present invention relates to an agent for the prevention and treatment of respiratory diseases such as antitussive expectoration and asthma, comprising a mixture of an ivy leaf extract and Coptidis rhizoma extract as an active ingredient, and a method for preparing the same. The mixtures exhibit markedly superior antitussive action, expectorant action, antihistamine action, inhibition of bronchial contractility, inhibition of airway hypersensitivity, suppression of bronchial obstruction and inhibition of inflammatory cell infiltration than each extract alone. Do it.

That is, the inventors of the present invention have a significantly increased antitussive effect, expectorant effect, antihistamine effect, bronchus in the case of a mixture of ivy leaf extract and rhubarb extract in a specific ratio compared to the case where the ivy leaf extract and the rhubarb extract are present alone. The present invention was confirmed to have a contraction inhibitory effect, an airway hyperresponsiveness inhibitory effect, a bronchial obstruction suppression effect, and an inflammatory cell infiltration inhibitory effect. will be.

Ivy belongs to the Araliaceae and means the plants of Hedera ( Hedera spp.), For example, Hedera algeriensis, Hedera azorica, Hedera canariensis, Hedera colchica, Hedera helix, Hedera hibernica, Hedera maderensis, Hedera nepalensis, Hedera pastuchowii, Hedera rhombea and the like, but is not limited thereto. In respiratory diseases, the therapeutic effects of dry ivy leaf extracts are due to the mucolytic, anticonvulsant and antitussive effects of glycoside saponins in the preparation. Mucolytic secretion of the extract is apparently due to the saponin properties of hederaglycoside, while the anticonvulsant effect of the inflammatory bronchus is parasympathetic neurosuppression of some glycosides. It is judged to be due to the effect. The antitussive effect of the extract is based on a mild soothing effect as a central inhibitory mechanism by having a special effect on the cough central. While having this effect, the extract has the advantage of not causing damage to the respiratory control center. In addition, there are no reports of side effects occurring in combination with ivy leaf extract and other preparations.

On the other hand, Huang Yan ( Coptidis rhizoma ) is a perennial herb belonging to Ranunculus family and is known to have pharmacological effects such as antimicrobial action, hypotensive action and anti-inflammatory action. It is used as a bladder, bronchial and nervous system agent. It is known to contain alkaloids such as berberine and magnoflorine, mainly as pharmacological components (Information Interpretation, Shin Min-kyo Bridge, Dohae Hyangdae (Medicinal Medicine), 490-493, 1990).

As described above, the present invention has antitussive action, expectorant action, antihistamine action, inhibitory action of bronchial contraction, inhibition of airway hypersensitivity, inhibition of bronchial blockage of lung tissue and inhibition of inflammatory cell infiltration in a mixture of ivy leaf extract and rhubarb extract. It is an object of the present invention to provide a mixture and a method of producing an optimal combination ratio in which the pharmacologically active synergistic effect is greatest by confirming the increase in the ivy leaf extract alone or the sulfur extract alone.

First, the present invention relates to a mixed extract having an antitussive expectorant action, an airway hyperresponsiveness inhibitory effect, a bronchial contraction inhibitory action, a bronchial obstruction inhibitory effect and an inflammatory cell infiltration inhibitory action containing a mixed extract of an ivy leaf and a yellow lotus.

In the present invention, the term 'mixed extract of ivy leaf and yellow lotus' refers to a mixture of ivy leaf extract and yellow leaf extract obtained by extracting ivy leaf and yellow lotus, respectively, and an extract obtained by extracting a mixture of ivy leaf and yellow lotus. Used to mean. In addition, the 'extract' refers to a crude extract or a specific solvent soluble extract (fraction), and may be in a solution, concentrate or powder state.

The present invention by using a mixed extract of ivy leaf and yellow lotus, antitussive action, expectorant action, antihistamine effect, airway hypersensitivity inhibitory effect, bronchial contraction inhibitory effect, bronchial blockage suppression of lung tissue than ivy leaf extract and yellow lotus extract alone And synergistic effect in inhibiting inflammatory cell invasion, antitussive expectoration, antihistamine, bronchial contraction inhibition, airway hyperresponsiveness suppression, bronchial obstruction suppression by ivy leaf extract and rhubarb extract. And in order to maximize the synergistic effect of the inflammatory cell invasion inhibitory activity, the content ratio of the ivy leaf extract and the sulfur extract in the mixed extract is 0.1: 1 to 10: 1 based on the weight of the solid content Same), preferably 0.2: 1 to 5: 1, more preferably 1: 1 to 4: 1, More preferably 1.5: 1 to 3.5: 1, most preferably 2.5: 1 to 3.5: 1. The 'solid content' means a state in which the solvent used to prepare the extract removed.

In one embodiment, the mixed extract according to the present invention may be a mixture of ivy leaf extract and sulfur extract. The ivy leaf extract is a crude extract obtained by extracting the dried ivy leaf with water and at least one selected from the group consisting of 1 to 4 carbon atoms or a straight chain or branched alcohol, or a lower alcohol solution containing 1 to 6 carbon atoms in the crude extract, Preferably, the solvent-soluble extract obtained by adding one or more selected from the group consisting of aqueous propyl alcohol solution, isopropyl alcohol aqueous solution, and saturated butyl alcohol. The ivy used to manufacture the ivy leaf extract is Hedera spp. For example, Hedera algeriensis, Hedera azorica , Hedera canariensis , Hedera colchica , Hedera helix, Hedera hibernica, Hedera maderensis , Hedera nepalensis , Hedera pastuchowii , Hedera rhombea may be one or more selected from the group consisting of, but is not limited thereto.

In addition, the extract of the yellow lotus is a yellow lotus ( Coptidis rhizoma ), preferably crude extract obtained by extracting the root of the root of rhizome into at least one selected from the group consisting of water and linear or branched alcohols having 1 to 4 carbon atoms or an aqueous solution of lower alcohol having 1 to 6 carbon atoms in the crude extract. , Preferably a solvent-soluble extract obtained by adding one or more selected from the group consisting of aqueous propyl alcohol and aqueous isopropyl alcohol, and saturated butyl alcohol.

In a preferred embodiment, the solvent used for the preparation of crude extracts of ivy leaf or rhubarb is water or 10 to 70% (v / v), preferably 20 to 60% (v / v), more preferably about It may be at least one member selected from the group consisting of 25 to 55% (v / v) linear or branched alcohol solution of 1 to 4 carbon atoms, preferably an aqueous methanol solution, an aqueous ethanol solution, and a saturated butyl alcohol.

In addition, the solvent used in the preparation of the solvent-soluble extract of the ivy leaf or the yellow lotus is 10 to 70% (v / v), preferably 20 to 60% (v / v), more preferably about 25 to 55% (v / v) may be one or more selected from the group consisting of a lower alcohol aqueous solution of 1 to 6 carbon atoms, preferably a propyl alcohol aqueous solution, an isopropyl alcohol aqueous solution, and a saturated butyl alcohol.

In another embodiment, the mixed extract according to the present invention may be an extract of a mixture of ivy leaf and rhubarb. For example, the mixed extract is mixed with the ivy leaf and the yellow lotus, and then to the crude extract or crude extract obtained by extracting the mixture with at least one selected from the group consisting of water and linear or branched alcohol having 1 to 4 carbon atoms It may be a solvent-soluble extract obtained by adding one or more selected from the group consisting of a lower alcohol aqueous solution of 1 to 6 carbon atoms, preferably an aqueous propyl alcohol solution and an isopropyl alcohol solution, and a saturated butyl alcohol.

In a preferred embodiment, the solvent used for the preparation of the crude extract of the ivy leaf and the sulfur mixture is water or 10 to 70% (v / v), preferably 20 to 60% (v / v), more preferably About 25-55% (v / v) aqueous methanol solution, ethanol aqueous solution or saturated butyl alcohol. In addition, the solvent used to prepare the solvent-soluble extract is 10 to 70% (v / v), preferably 20 to 60% (v / v), more preferably about 25 to 55% (v / v) It may be one or more selected from the group consisting of a lower alcohol aqueous solution of 1 to 6 carbon atoms, preferably a propyl alcohol aqueous solution, an isopropyl alcohol aqueous solution, and a saturated butyl alcohol.

In another aspect, the present invention relates to a method for producing a mixed extract of ivy leaf and rhubarb with antitussive expectoration, antihistamine, bronchial contraction suppression, airway hypersensitivity suppression, bronchial obstruction of pulmonary tissue and inhibitory activity of inflammatory cell infiltration.

In one embodiment, the preparation method comprises at least one selected from the group consisting of water and straight or branched alcohols having 1 to 4 carbon atoms, such as water, or 10 to 70% (v / v), Extracting preferably 20 to 60% (v / v), more preferably about 25 to 55% (v / v) aqueous methanol solution, aqueous ethanol solution or saturated butyl alcohol to obtain an ivy leaf extract;

At least one selected from the group consisting of water, and straight or branched alcohols having 1 to 4 carbon atoms, such as water, or 10 to 70% (v / v), preferably 20 Extracting to 60% (v / v), more preferably about 25 to 55% (v / v) aqueous methanol solution, aqueous ethanol solution or saturated butyl alcohol to obtain a sulfur extract; And the obtained ivy leaf extract and the rhubarb extract based on a solids weight of 0.1: 1 to 10: 1 (the ivy leaf extract weight: the weight of the sulfur extract), preferably 0.2: 1 to 5: 1, more preferably 1: 1. To 4: 1, more preferably 1.5: 1 to 3.5: 1, most preferably 2.5: 1 to 3.5: 1.

At this time, each of the ivy leaf extract manufacturing step and the manufacturing step of the yellow lotus extract is selected from the group consisting of a lower alcohol aqueous solution of 1 to 6 carbon atoms, preferably saturated butyl alcohol, propyl alcohol and isopropyl alcohol in each extract. The method may further include purifying the extract by taking a solvent-soluble extract obtained by adding more than one species.

In the above embodiment, the manufacturing process of the ivy leaf extract according to the present invention will be described in more detail as follows: After cutting the ivy leaf and washing with water to remove the constriction and drying, the weight of the dried ivy leaf At least one selected from the group consisting of about 5 to 20 volume times, preferably 7 to 15 volume times water, and straight or branched alcohols having 1 to 4 carbon atoms, such as water or 10 to 70% (v / v), preferably 20-60% (v / v), more preferably about 25-55% with reflux extraction with aqueous methanol, aqueous ethanol or saturated butyl alcohol. Extraction temperature is 40 to 110 ℃, preferably 55 to 90 ℃.

  After extraction, the filtrate is collected by filtration and at least one selected from the group consisting of about 5 to 15 volumes of water, preferably 8 to 12 volumes of water, and straight or branched alcohols having 1 to 4 carbon atoms, For example, water, or 10 to 70% (v / v), preferably 20 to 60% (v / v), more preferably about 25 to 55% (v / v) aqueous methanol solution, aqueous ethanol solution or water Extracted with reflux with saturated butyl alcohol. The extraction temperature is not particularly limited but is preferably 40 to 110 ° C, preferably 55 to 90 ° C. After extraction, filtration and combined with the filtrate previously obtained to concentrate under reduced pressure to prepare an ivy leaf extract. As such, the extraction efficiency can be increased by mixing the filtrate obtained after the extraction and the extraction over each of the two times, but the extract of the present invention is not limited to the number of extraction.

When the amount of the solvent used in the preparation of the ivy leaf extract is too small, the stirring becomes difficult, the solubility of the extract is lowered, the extraction efficiency is lowered, if too large, the amount of lower alcohol used in the next purification step is more economical Since this may cause problems in handling, it is better to use the solvent in the above range.

 In a preferred embodiment of the present invention, a method of re-extracting after primary extraction may be adopted, which is a loss due to the high water content of the herbal medicine itself even if it is effectively filtered in the case of mass production of herbal extracts. Extraction alone is to reduce the extraction efficiency 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 multistage extraction is not economical.

The concentrate thus obtained is two to three times with about 10-30 weight times of the total amount of the concentrate, preferably about 15-25 weight times, in order to control the content of the remaining lower alcohol to be suitable for use as a pharmaceutical ingredient. It can be prepared as an ivy leaf extract in powder form by azeotropically concentrating, adding the same amount of water, and then suspending it homogeneously and lyophilizing.

The manufacturing process of the sulfur extract according to the present invention is described as follows: After cutting the root portion of the sulfur extract (Coptidis rhizoma), 3 to 20 vol., Preferably 5 to 15 vol. And at least one selected from the group consisting of linear or branched alcohols having 1 to 4 carbon atoms, such as water or 10 to 70% (v / v), preferably 20 to 60% (v / v), more Preferably, about 25-55% (v / v) aqueous methanol solution, ethanol aqueous solution or saturated butyl alcohol are added and refluxed for 1 to 10 hours, preferably 2 to 5 hours. The extraction temperature is not particularly limited but is preferably 40 to 110 ° C, preferably 55 to 90 ° C. After extraction, the filtrate is collected by filtration, and at least one selected from the group consisting of 1 to 10 vol. Of water, preferably 4 to 7 vol. Of water, and straight or branched alcohol having 1 to 4 carbon atoms to the residue. For example, water or 10-70% (v / v), preferably 20-60% (v / v), more preferably about 25-55% (v / v) aqueous methanol solution, aqueous ethanol solution or Saturated butyl alcohol can be added and warmed to re-extract for 1 to 10 hours, preferably 2 to 5 hours, filtered, mixed with the previous filtrate and concentrated under reduced pressure to obtain a sulfur extract.

 As such, the extraction efficiency can be increased by mixing the filtrate obtained after the extraction and the extraction over each of the two times, but the extract of the present invention is not limited to the number of extraction.

When the amount of the solvent used in the preparation of the sulfur extract is too small, the stirring becomes difficult, the solubility of the extract is lowered, the extraction efficiency is lowered, if too large, the amount of lower alcohol used in the next purification step is more economical Since the problem may occur in handling, the amount of the solvent to be used is preferably within the above range.

 In a preferred embodiment of the present invention, a method of re-extracting after primary extraction may be adopted, which is a loss due to the high extractant content of the herbal medicine itself even if it is effectively filtered in the case of mass production of herbal extracts. Extraction alone is to reduce the extraction efficiency 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 multistage extraction is not economical.

After filtering and concentrating the sulfur extract obtained by extracting with methanol aqueous solution or ethanol aqueous solution or saturated butyl alcohol as described above, and then purifying the impurities such as unnecessary proteins, polysaccharides and fatty acids contained in the filtrate, Purifying the impurities may be further carried out by layer separation 1 to 10 times, preferably 2 to 4 times, with the same amount of lower alcohol to obtain a solvent fraction. At this time, a linear or branched alcohol having 1 to 6 carbon atoms may be used as the lower alcohol, preferably at least one selected from the group consisting of butyl alcohol, propyl alcohol and isopropyl alcohol. When the amount of the lower alcohol is less than that of the filtrate, fine particles are formed by unnecessary components such as fatty acids, and thus the layer separation is not smooth and the extraction content of the active ingredient is not efficient.

The lower alcohol fraction obtained after layer separation is concentrated under reduced pressure at 50 to 60 ° C. to remove the solvent remaining in the sample.

The concentrate thus obtained is about 10 to 30 times, preferably 15 to 25 times, more preferably about 20 weight times the total amount of the concentrate in order to adjust the content of the remaining lower alcohol to be suitable for use as a pharmaceutical raw material. 1 to 5 times, preferably 2 to 3 times azeotropically concentrated with water, and the same amount of water is added again to homogeneously suspend and then lyophilized to prepare as a powdery sulfur extract.

The first step of dividing the sulfur extract obtained above into a mixed solvent of methylene chloride and methanol (30: 1 to 7: 1 (v / v)) by silica gel column chromatography and dividing it into five small fractions, the first Components contained in the extract of the present invention were identified through a second step process of performing high performance liquid chromatography (HPLC) on the fractions of the step. That is, as a result of analyzing the components of the extracts of the present invention, it was confirmed that berberine, palmatin, copticin, columamine, and jathrorizine are included.

The barberry extract of the present invention includes berberine, palmin, copticin, columamine, and jathrorizine, preferably the composition comprises 15 to 30 parts by weight of berberine, 4 to 10 parts by weight of palminth, and 4 to 4 incorporation. 10 parts by weight, columamine 0.3 to 5 parts by weight, it may be to include 0.3 to 5 parts by weight of the jathrorizine.

Referring to the process of analyzing the components contained in the sulfur extract is as follows.

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, palmtin, and copticin were commercially available from Sigma, and columamine and jathrorizine were used after separation and purification from rhubarb. The mobile phase is 0 minutes to 60 minutes (A: B = 9: 1 to 6: 4) and 60 minutes 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.

Referring to the manufacturing process of the mixed composition for the two extracts prepared as described above are as follows.

The ivy leaf extract and the rhubarb extract obtained above are 0.1: 1 to 10: 1 based on the solids weight (the ivy leaf extract weight: the weight of the yellow leaf extract), preferably 0.2: 1 to 5: 1, more preferably 1: 1. To 4: 1, more preferably 1.5: 1 to 3.5: 1, most preferably 2.5: 1 to 3.5: 1.

In order to homogeneously mix each extract in the mixed extract, about 2 to 3 weight times of water was added to the mixed extract, and then concentrated under reduced pressure at a temperature of 50 to 60 ° C, and the same amount of water was added to the concentrate. The powder composition may be prepared by adding again, suspending homogeneously and lyophilizing.

In another embodiment, the preparation method is 1: 4 to 7: 1 (Ivy leaf weight: weight of sulfur leaf), preferably 1: 1 to 6, based on the weight of the dry ivy leaf and the yellow lotus, preferably the root of the yellow lotus. 1: 1, more preferably 2: 1 to 5: 1, to prepare a mixture of ivy leaf and yellow lotus; And at least one selected from the group consisting of water and linear or branched alcohols having 1 to 4 carbon atoms, such as water, or 10 to 70% (v / v), preferably 20 to 60% (v) in the mixture. / v), more preferably about 25 to 55% (v / v) may be extracted with an aqueous methanol solution, an ethanol aqueous solution or a saturated butyl alcohol to obtain an extract of the mixture of ivy leaf and sulfur.

The method is one selected from the group consisting of linear or branched lower alcohol aqueous solution of 1 to 6 carbon atoms, preferably propyl alcohol, isopropyl alcohol, and saturated butyl alcohol after the step of obtaining an extract of the mixture of ivy leaf and sulfur. The method may further include purifying the extract by taking a solvent-soluble extract obtained by adding more than one species.

Specific details of the crude extract and the solvent-soluble extract preparing step are as described above to prepare each herbal extract.

The extraction method used in the present invention may be all methods commonly used, and for example, cold extraction, hot water extraction, ultrasonic extraction, or reflux cooling extraction method, but is not limited thereto.

In another aspect, the present invention provides an antitussive composition containing a mixed extract of the above ivy leaf extract and sulfur extract as an active ingredient. In addition, the present invention provides an expectorant composition containing a mixed extract of the above-described ivy leaf extract and sulfur extract as an active ingredient.

In addition, the mixed extract of the ivy leaf extract and the rhubarb extract according to the present invention is elevated antitussive action, expectorant action, antihistamine action, bronchial contraction inhibitory action, airway hypersensitivity action, bronchial obstruction suppression and inflammation than the respective extract alone Since the cell invasion inhibitory effect, it can be usefully used for the prevention, treatment or symptom relief of the associated respiratory diseases. Accordingly, the present invention provides a composition for the prevention or treatment of respiratory diseases, which contains a mixed extract of ivy leaf extract and sulfur extract as an active ingredient.

The respiratory disease can be any disease that accompanies cough and sputum, eg, general cough or sputum; Emphysema with cough or sputum; Bronchitis, such as chronic bronchitis, acute bronchitis, catarrhal bronchitis, obstructive or inflammatory bronchial disease; Asthma, such as bronchial asthma, atopic asthma, atopic bronchial IgE mediated asthma, non-atopic asthma, allergic asthma, non-allergic asthma; Chronic or acute bronchial contraction; Wheezing infant syndrome; Chronic obstructive pulmonary disease; Bronchial adenoma; Isolated pulmonary nodules; tuberculosis; Empyema; Lung abscess; cold; flu; And it may be one or more diseases selected from the group consisting of histocyte hyperplasia, etc.

The content of the mixed extract as an active ingredient in the composition according to the present invention can be appropriately adjusted according to the use form and purpose, patient condition, type and severity of symptoms, etc., based on the solid content weight of 0.001 to 99.9% by weight, preferably 0.1 to It may be 50% by weight, but is not limited thereto.

The composition according to the invention can be administered to a mammal, including humans, by various routes. The mode of administration can be any of the routinely used methods and can be administered, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection. The compositions of the present invention are formulated in oral dosage forms, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, parenteral formulations in the form of transdermal agents, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Can be used.

The composition of the present invention may further contain an adjuvant such as a pharmaceutically suitable and physiologically acceptable carrier, excipient and diluent in addition to the mixed extract. Carriers, excipients and diluents that may be included in the compositions of the present invention 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 can be 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 can be 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, transdermal agents and the like. 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.

In an embodiment in which the composition of the present invention is applied to a human, the herbal extract composition of the present invention may be administered alone, but generally with a pharmaceutical carrier selected in consideration of the mode of administration and standard phamaceutical practice. May be administered in admixture. For example, the mixed extract-containing compositions of the present invention may be in the form of tablets containing starch or lactose, or in the form of capsules containing singly or excipients, or elixirs or suspensions containing chemicals that flavor or color. Oral, oral or sublingual. Such liquid preparations may be suspending agents (e.g., semisynthetic glycerides such as methylcellulose, witepsol or mixtures of apricot kernel oil and PEG-6 esters or PEG-8 and caprylic / capric Pharmaceutically acceptable additives such as glyceride mixtures such as mixtures of glycerides).

The dosage of the mixed extract-containing composition of the present invention may vary depending on the age, weight, sex, dosage form, health condition and degree of disease of the patient, and may be once or a day at a predetermined time interval according to the judgment of a doctor or pharmacist. Divided doses may also be administered. For example, the daily dosage may be 0.5 to 500 mg / kg, preferably 1 to 300 mg / kg, based on the active ingredient content. The above dosages are illustrative of the average case and may be high or low depending on individual differences. If the daily dose of the mixed extract-containing composition of the present invention is less than the dose, a significant effect cannot be obtained, and if it exceeds the above, it is not only uneconomical but also out of the range of normal doses, which may cause undesirable side effects. Since it may occur, it is preferable to set the above range.

In another aspect, the present invention provides a health functional food for preventing or ameliorating respiratory diseases such as antitussive, expectorant and asthma, containing a mixed extract of ivy leaf and yellow lotus. The health functional food may be various foods, beverages, food additives and the like.

The content of the mixed extract as an active ingredient contained in the health functional food is not particularly limited depending on the form of the food, desired use, etc., for example, can be added to 0.01 to 15% by weight of the total food weight, health beverage composition It can be added in a ratio of 0.02 to 10 g, preferably 0.3 to 1 g based on 100 ml of silver.

 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 conventional monosaccharides such as disaccharides such as glucose and fructose, 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.

In particular, when the herbal composition of the present invention is administered to the human body, in view of the general characteristics of natural extracts, it is considered that there is no concern of side effects compared with other synthetic pharmaceuticals, and in fact, the toxicity test results of the standardized herbal composition have no effect on the living body. Proved to be absent.

The mixture of ivy leaf extract and rhubarb extract according to the present invention is ivy leaf extract and yellow leaf extract in expectorant activity, antitussive activity, antihistamine activity, bronchial contraction inhibitory activity, airway hypersensitivity inhibitory activity, bronchial obstruction inhibition and inflammatory cell infiltration inhibitory activity, etc. The effect is superior to that of the extract alone. In other words, by mixing with the extract of ivy leaves in the optimum ratio of the ivy leaf extract currently widely used as an antitussive expectorant, antihistamine activity, bronchial contraction inhibitory activity, airway hypersensitivity inhibitory activity, lungs Inhibition of bronchial blockage and inflammatory cell invasion activity of tissues may increase the indications for not only ginseng sputum but also a wide range of respiratory diseases such as asthma and bronchitis.

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

Example 1: Preparation of Raw Material Extract

Reference Example 1: Preparation of Ivy Leaf Extract

The leaves of the ivy ( Hedera helix ) were washed with water to remove impurities and allowed to dry completely. 3 l of 30% (v / v) ethanol aqueous solution was added to 250 g of the ivy leaf thus prepared, reflux-cooled for 6 hours, filtered and the filtrate was collected, and 2.5 l of 30% (v / v) ethanol aqueous solution was added to the residue. The filtrate obtained by re-warming extraction at 80 ° C. for 3 hours was mixed with the previously collected filtrate and concentrated under reduced pressure. After azeotropically concentrating azeotropically by adding 0.2 L of water in most of the solvents evaporated, repeating twice, and homogeneously suspending again with the same amount of water, and then lyophilized to obtain 42.4 g of powdery ivy leaf extract. .

Reference Example 2: Preparation of Rhubarb Extract

Coptidis rhizoma was washed with water to remove contaminants and allowed to dry completely. 250 g of the yellow soybean prepared as described above was extracted under reflux with 2.0 L of 50% (v / v) ethanol solution twice at 80 ° C. for 3 hours, and the extract was filtered and concentrated under reduced pressure to obtain 57.5 g of crude extract, which was 0.3 L Water was added and suspended, 0.6L of saturated butyl alcohol was added thereto, and the layers were separated twice, and only the butyl alcohol fractions were collected and concentrated under reduced pressure until drying. Most of butyl alcohol and water were evaporated, 0.2 L of water was added, the azeotropic concentration was repeated, and then repeated twice. Finally, the same amount of distilled water was added and suspended, followed by freeze-drying to obtain 31.8 g of powdered sulfur extract.

[Example 2: Preparation of a mixed extract of the ivy leaf extract and the yellow lotus extract]

Preparation Example 1 Mixed Extract (0.2: 1)

The ivy leaf extract of Reference Example 1 and the rhubarb extract of Reference Example 2 were mixed at a weight ratio of 0.2: 1 (Ivy leaf extract weight: weight of the sulfur extract). In order to achieve a homogeneous mixture, 2-3 weight times of water was added to the mixed extract, concentrated under reduced pressure at a temperature of 50 to 60 ° C., and the same amount of water was added again to the obtained concentrate, and then suspended homogeneously. By lyophilization, a powdered extract was prepared.

Preparation Example 2: Mixed Extract (0.4: 1)

Mixing in the same manner as in Preparation Example 1, except that the mixing ratio of the ivy leaf extract of Reference Example 1 and the yellow lotus extract of Reference Example 2 to 0.4: 1 (Ivy leaf extract weight: weight of the yellow leaf extract) based on the weight. An extract was prepared.

Preparation Example 3: Mixed Extract (1: 1)

The mixing ratio of the ivy leaf extract of Reference Example 1 and the sulfur extract of Reference Example 2 is mixed in the same manner as in Preparation Example 1, except that the mixing ratio is 1: 1 (the ivy leaf extract weight: the weight of the yellow extract). An extract was prepared.

Preparation Example 4 Mixed Extract (1.5: 1)

The mixing ratio of the ivy leaf extract of Reference Example 1 and the yellow lotus extract of Reference Example 2 was mixed in the same manner as in Preparation Example 1, except that the mixing ratio was 1.5: 1 (the ivy leaf extract weight: the weight of the sulfur extract). An extract was prepared.

Preparation Example 5 Mixed Extract (2: 1)

Mixing in the same manner as in Preparation Example 1, except that the mixing ratio of the ivy leaf extract of Reference Example 1 and the yellow lotus extract of Reference Example 2 is 2: 1 (weight of the ivy leaf extract: weight of the yellow leaf extract). An extract was prepared.

Preparation Example 6 Mixed Extract (2.5: 1)

The mixing ratio of the ivy leaf extract of Reference Example 1 and the yellow lotus extract of Reference Example 2 was mixed in the same manner as in Preparation Example 1, except that the mixing ratio was 2.5: 1 (the ivy leaf extract weight: the weight of the sulfur extract). An extract was prepared.

Preparation Example 7 Mixed Extract (3: 1)

Mixing in the same manner as in Preparation Example 1, except that the mixing ratio of the ivy leaf extract of Reference Example 1 and the yellow lotus extract of Reference Example 2 is 3: 1 (Ivy leaf extract weight: weight of the yellow leaf extract) based on the weight. An extract was prepared.

Preparation Example 8 Mixed Extract (3.5: 1)

The mixing ratio of the ivy leaf extract of Reference Example 1 and the yellow lotus extract of Reference Example 2 was mixed in the same manner as in Preparation Example 1, except that the mixing ratio was 3.5: 1 (the ivy leaf extract weight: the weight of the sulfur extract). An extract was prepared.

Preparation 9: Mixed Extract (4: 1)

The mixing ratio of the ivy leaf extract of Reference Example 1 and the yellow lotus extract of Reference Example 2 is mixed in the same manner as in Preparation Example 1, except that the mixing ratio is 4: 1 (the ivy leaf extract weight: the weight of the sulfur extract). An extract was prepared.

Preparation Example 10 Mixed Extract (4.5: 1)

Mixing in the same manner as in Preparation Example 1, except that the mixing ratio of the ivy leaf extract of the Reference Example 1 and the yellow lotus extract of the Reference Example 2 to 4.5: 1 (Ivy leaf extract weight: weight of the yellow leaf extract). An extract was prepared.

Preparation Example 11 Mixed Extract (5: 1)

Mixing in the same manner as in Preparation Example 1, except that the mixing ratio of the ivy leaf extract of the Reference Example 1 and the yellow lotus extract of the Reference Example 2 to 5: 1 (weight of the ivy leaf extract: weight of the sulfur extract). An extract was prepared.

Example 3: Pharmacological Activity Test

Experimental Example 1: expectoration activity test

In order to evaluate the expectorant activities of the mixed extracts prepared in Preparation Examples 1 to 11, and Reference Example 1 and Reference Example 2, which are each single extracts, 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.) Evaluation experiments were performed, and the results are shown in FIG. In FIG. 1, the positive control group (Ambroxol) was orally administered at 250 mg / kg and the remaining test drug at 500 mg / kg. In addition, to evaluate the expectorant activity for each dose for the preparation 7 of the most excellent activity of the results are shown in Table 1 below.

Experimental Method

A positive control drug (Ambroxol, ambroxol, Sigma) and test drug were orally administered to male mice (8 weeks old, Samtaco BioKorea), and phenol red was intraperitoneally injected (500 mg / kg dose). Phenol red was dissolved in physiological saline). After 30 minutes, anesthetized with diethyl ether, the abdominal aorta was cut and bled, and the entire trachea was excised. The separated organs were placed in 1 ml of physiological saline and washed for 30 minutes, and 1 N caustic soda (NaOH) was added to the supernatant obtained by centrifugation at 10,000 rpm for 5 minutes at room temperature (0.1 ml of 1 N NaOH per 1 ml of supernatant). It was. The absorbance at 546 nm was measured for these and the expectorant activity was evaluated as the concentration of phenol red discharged.

[Experiment result]

The sputum discharge capacity according to the administration dose of the amproxol used as the positive control drug and the material obtained in Preparation Example 7 is shown in Table 1 below.

division Dosage (mg / kg) Sputum discharge capacity (%) Preparation Example 7 50 17 100 30 150 38 200 44 500 56 Positive control group Ambroxol 250 34

As can be seen in Table 1, the material of Preparation Example 7 according to the present invention was shown to have a better sputum discharge capacity than the positive control drug.

In addition, the expectorant effect of the ivy leaf extract of Reference Example 1, the yellow lotus extract of Reference Example 2, and the mixed extract of Preparation Examples 1 to 11 was measured and shown in FIG. 1. As shown in FIG. 1, the ivy leaf extract of Reference Example 1, the yellow lotus extract of Reference Example 2, and the mixed extract of Preparation Examples 1 to 11 were confirmed to have an expectorant effect. Moreover, in all preparations, the extract of Reference Example 1 and Reference Example 2 showed an enhanced expectoration effect than when used alone, and the pharmacologically active synergistic effect of the mixed composition according to the present invention can be confirmed from the above results. In particular, in the case of Preparation Examples 3 to 9 shows an excellent expectorant effect than the conventional expectorant Ambroxol used as a positive control group, when the ivy leaf extract and the rhubarb extract are mixed in a weight ratio of 1: 1 to 4: 1 by weight It can be seen that it shows a better elevated expectoration effect. In addition, Table 1 above was evaluated by the most active Preparation Example 7 by the dose, showing an effect comparable to the positive control group at a dose of 100mg / kg or more, in particular 150 mg / kg or more shows an effect better than the positive control group It can be seen.

Experimental Example 2: Antitussive Activity Evaluation Test

In order to measure the antitussive activity of the mixed extracts prepared according to Preparation Examples 1 to 11, and Reference Example 1 and Reference Example 2, which are single extracts of each, 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.) 2 is shown. In FIG. 2, the positive control group (Theobromine, Sigma) was orally administered 50 mg / kg, and the remaining test drug 200 mg / kg. Among them, the antitussive activity of each of the most excellent Preparation Example 7 was evaluated by the dose, and the results are shown in Table 2 below.

[Experimental method]

After 1 hour of oral administration of the drug to male guinea pigs (6 weeks old, Samtaco BioKorea), the guinea pigs were placed in a plastismograph chamber (Buxco, U.S.A.), followed by spraying a cough inducing agent (citric acid) to induce cough. Theobromine (50 mg / kg), used as an antitussive agent, was used as a positive control group, and guinea pigs were exposed to 2 M citric acid for 10 minutes, and cough water generated at this time was measured for 15 minutes.

[Experiment result]

The antitussive activity of theobromine used as a positive control drug and the dose of the material obtained in Preparation Example 7 is shown in Table 1 below.

division Dosage (mg / kg) Jinhae inhibitory ability (%) Preparation Example 7 50 29 100 51 150 69 200 83 Positive control group Theobromine 50 56

In addition, the antitussive effect of the ivy leaf extract of Reference Example 1, the sulfur extract of Reference Example 2, and the mixed extract of Preparation Examples 1 to 11 is shown in FIG. As shown in FIG. 2, the ivy leaf extract of Reference Example 1, the sulfur extract of Reference Example 2, and the mixed extract of Preparation Examples 1 to 11 were all confirmed to have antitussive effect. Moreover, in all preparations, the extracts of Reference Example 1 and Reference Example 2 exhibited enhanced antitussive effects than those used alone, and the pharmacologically active synergistic effects of the mixed compositions according to the present invention can be confirmed from the above results. In particular, in the case of Preparation Examples 3 to 9 shows an excellent antitussive effect than the existing antitussive agent theobromine used as a positive control group, when the ivy leaf extract and the rhubarb extract are mixed in a weight ratio of 1: 1 to 4: 1 by weight, It can be seen that it shows an excellent elevated antitussive effect. In addition, Table 2 above was evaluated by the most active Preparation Example 7 by the dose, showing an effect comparable to the positive control group at a dose of 100mg / kg or more, in particular at 150 mg / kg or more shows an excellent effect than the positive control group It can be seen.

Experimental Example 3: Antihistamine Effect Test

In order to measure the antihistamine efficacy in mast cells for the mixed extracts prepared in Preparation Examples 1 to 11, and each of the single extracts, Honuchi et al. (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 results are shown in Table 3 below. In addition, the antihistamine efficacy was evaluated for each dose of Preparation Example 7 having excellent activity, and the results are shown in Table 4 below.

Experimental Method

Antihistamine effects were measured using celiac cells of male rats (7 weeks old, Samtaco BioKorea) sensitized with egg white albumin (Sigma). Ketotifen (Sigma) was used as a positive control. Rats sensitized with egg white albumin were orally administered with ketotifen at 5 mg / kg and test drug at a concentration of 200 mg / kg for 3 days, and then the abdominal mast cells were isolated. Ketotifen was isolated at 0.01 mg / ml, 0.1 mg / ml and 1.0 mg / ml in isolated celiac mast cells (2 x 10 ⁵ cells / ml). Treatment was at a concentration of 10 mg / ml. 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 test substances inhibited histamine release from mast cells. Relative amounts were determined relative to the amount of histamine released from mast cells of rats that received no drug.

[Experiment result]

division Oral Dose (mg / kg) Mast cell treatment (mg / ml) Histamine Release Rate (%) Preparation Example 1 200 0 48 0.1 42 1.0 39 10.0 32 Preparation Example 2 200 0 47 0.1 41 1.0 38 10.0 31 Preparation Example 3 200 0 41 0.1 37 1.0 34 10.0 28 Preparation Example 4 200 0 39 0.1 37 1.0 33 10.0 26 Preparation Example 5 200 0 37 0.1 34 1.0 31 10.0 25 Preparation Example 6 200 0 36 0.1 34 1.0 31 10.0 24 Preparation Example 7 200 0 34 0.1 32 1.0 29 10.0 20 Preparation Example 8 200 0 35 0.1 33 1.0 30 10.0 22 Preparation Example 9 200 0 38 0.1 35 1.0 31 10.0 27 Preparation Example 10 200 0 42 0.1 39 1.0 36 10.0 31 Preparation Example 11 200 0 49 0.1 44 1.0 39 10.0 30 Reference Example 1 200 0 45 0.1 40 1.0 35 10.0 34 Reference Example 2 200 0 39 0.1 33 1.0 32 10.0 25

division Oral Dose (mg / kg) Mast cell treatment (mg / ml) Histamine Release Rate (%) Positive control Ketotifen 5 0 44 0.01 41 0.1 32 1.0 19

division Oral Dose (mg / kg) Mast Cell Concentration (mg / ml) Histamine Release Rate (%) Preparation Example 7 100 0 49 0.1 39 1.0 35 10.0 34 200 0 34 0.1 32 1.0 29 10.0 20 400 0 21 0.1 18 1.0 17 10.0 17 Reference Example 1 200 0 45 0.1 40 1.0 35 10.0 34 Reference Example 2 200 0 39 0.1 33 1.0 32 10.0 25 Positive control Ketotifen 5 0 44 0.01 41 0.1 32 1.0 19

As shown in Table 3, the ivy leaf extract of Reference Example 1, the sulfur extract of Reference Example 2, and the mixed extract of Preparation Examples 1 to 11 according to the present invention can be confirmed that there is an antihistamine effect, more specifically, It can be seen that the synergistic effect of the activity is further enhanced when the extracts of Reference Example 1 and Reference Example 2 which individually exhibit antihistamine activity are mixed. In addition, Table 4 shows that the best anti-histamine activity by the evaluation of the most active Preparation Example 7 by dose.

Experimental Example 4: Bronchial contraction inhibition test (in vitro)

In order to measure the airway contraction inhibitory effect on the mixed extracts prepared in Preparation Examples 1 to 11, and each of the single extracts, using a method such as Casoni GL ( Clin Exp Allergy ., 33, 999-1004, 2003, Anesth Analg ., 89, 191-196, 1999, Br J Pharmacol ., 128, 577-584, 1999). The results are shown in FIG. In Figure 3, the positive control group (sodium nitroprusside dihydrate) was 2.6 x 10 ^-5 mg / ml, and the remaining test drug was 0.50 mg / ml. Is shown in Table 5 below.

Experimental Method

Hartely male guinea pigs (350-400 g, Daehan Biolink) were anesthetized with pentobarbital sodium (75 mg / kg) and anesthetized before bronchial extraction. In the liquid phase of Krebs-Henseleit, the extracted bronchus is cut into 3 ~ 5 mm, divided into segments, fixed in an organ bath, perfused with 10 ^-4 M histamine, and then contracted to remove the bronchial tube. Was measured. Relaxation rate of the test substance-treated group (%) by subtracting the relaxation rate of the excipient control group (%) calculated under the same conditions from the relaxation rate (%) calculated on the assumption that 10 ^-4 M histamine induced contraction force was 100% ) Was calculated.

[Experiment result]

The bronchial contraction inhibitory activity according to the concentration of SNP (sodium nitroprusside dihydrate, Sigma) used as a positive control drug and the test substance obtained in Preparation Example 7 is shown in Table 5 below.

division Concentration (mg / ml) Bronchial Relaxation Rate (%) Preparation Example 7 0.25 20 0.50 48 1.00 83 Positive control group SNP 2.6 x 10 ^-5 30

In addition, the bronchial contraction inhibitory effect of the ivy leaf extract of Reference Example 1, the yellow lotus extract of Reference Example 2, and the mixed extract of Preparation Examples 1 to 11 is shown in FIG. As shown in FIG. 3, the ivy leaf extract of Reference Example 1, the yellow lotus extract of Reference Example 2, and the mixed extract of Preparation Examples 1 to 11 were confirmed to have bronchial contraction inhibitory effects. Moreover, in all preparations, the extracts of Reference Example 1 and Reference Example 2 showed an enhanced bronchial contraction inhibitory effect than when used alone. It can be seen that it exhibits shrinkage inhibiting activity.

Experimental Example 5: Anti-asthma in vivo effect test using mouse asthma model

In order to confirm the anti-asthma effect using the mouse asthma model of Preparation Example 7 according to the results of Experimental Example 1, Tang M.L.K. (Pulmonary Pharmacology & Therapeutics, 14, 203-210, 2001, Immunology and Cell Biology 79, 141-144, 2001, Journal of Experimental Medicine 189 (10), 1621-1629, 1999). Evaluation items included airway hyper-responsiveness (AHR) survey, IgE levels in plasma, cell number and immune cell distribution in bronchoalveolar lavage fluid (BALF), lung Histological observation, RT-PCR (IL-1β, IL-4, IL-13r2a) was confirmed.

Experimental Method

Balb / c mice (18-21 g, Orient Bio, Inc.) were intraperitoneally administered 500 ul (20 ug as OVA) per day of a solution prepared for intraperitoneal injection of ovalbumin (OVA, Sigma) on Day 0 and Day 7. The solution prepared for OVA inhalation was inhaled using nubulizer (NE-U17, OMRON Co. Ltd, Japan) and then inhaled 5% OVA solution prepared for 30 minutes daily from Day 14 to Day 20. As a positive control agent, 30 mg / kg of montelukast sodium and 200 mg / kg of all test substances were used. For positive control drugs and test substances, sensitized OVA inhalation for 7 days from Day 14 to Day 20 After oral administration daily 1 hour before the test was conducted according to the following evaluation items.

1) Investigation of Airway hyper-responsiveness (AHR) to methacholine

24 hours after the last OVA inhalation (Day 21), mice were placed in whole body plethysmograph (BUXCO, USA), 0.4 ml of methacholine 0, 10 and 20 mg / ml solutions were added to the aerosol and inhaled for 3 minutes each. Penh was measured for 4 minutes from the time of administration and the average value was taken as the Penh (enhanced pause) value at the corresponding methacholine dose.

2) Examination of IgE Levels in Plasma (Day 21)

After the measurement of 1), the animals were obtained by orbital blood collection, about 0.05 ml of blood was centrifuged, and the obtained plasma was subdivided into two portions and diluted 1:10 to 1 g, followed by IgE enzyme linked immunosorbent assay (ELISA; 151, SHIBAYAGI Co., Ltd., JAPAN) to investigate the level of immunoglobulin E (IgE) in plasma.

3) Cell number and immune cell distribution in bronchoalveolar lavage fluid (BALF) (Day 22)

After the blood collection of 2), animals were intraperitoneally injected with pentotalsodium (50mg / kg), and the left lung was fixed with forceps, and then injected into the bronchus and lungs using 0.4 ml of PBS solution, and the solution was taken out. This was repeated three times and centrifuged at 1,500 g to discard the supernatant and resuspended by adding 0.2 ml of PBS to the precipitate. The number of total leukocytes, neutrophils, eosinophils, neutrophils and lymphocytes was measured using an automated hematology analyzer (Advia 120 coulter counter, BAYER, Germany).

4) Observation of lung tissue

The left lung fixed with forceps was removed and fixed in 10% formalin solution for one day. Paraffin-infiltrated tissues were cut into 3-4 um thickness using a microtome to prepare tissue sections. The resulting slides were stained with Eosin and covered with a cover slide with balsam to observe the degree of occlusion of bronchioles and the infiltration of inflammatory cells.

5) RT-PCR

The right lung tissue was quickly removed and stored at -80 ° C until the test was run. The RNA was isolated from the tissue using RNA-Bee (Tel-Test Inc. USA) and the isolated RNA was quantified at 260 nm using a spectrophotometer, and 4 ug of RNA, Oligo dT 2 ul (Promega), and DEPC water were Add 10 ul and react for 5 minutes at 65 ℃. Add 5x reaction buffer 5 ul + RTase 2ul + dNTP 5 ul + RNase inhibitor 1 ul + DEPC water 2 ul, and add a total of 25 ul at 42 ℃ for 1 hour. The reaction was carried out at 100 ° C. for 5 minutes to make cDNA. The reagents used were all purchased from Promega. It was PCR with the prepared primers (IL-1β, IL-4, IL-13r2a). The primers used are as follows:

IL-1β forward: 5'-TCATGGGATGATGATGATAACCTGCT-3 '(SEQ ID NO: 1),

IL-1β reverse: 5'-CCCATACTTTAGGAAGACACGGATT-3 '(SEQ ID NO: 2);

IL-4 forward: 5'-TCATCGGCATTTTGAACGAG-3 '(SEQ ID NO: 3),

IL-4 reverse: 5'-GAATCCAGGCATCGAAAAGC-3 '(SEQ ID NO: 4);

IL-13r2a forward: 5'-GGTTATGCCAAATGCACTTGAG-3 '(SEQ ID NO: 5),

IL-13r2a reverse: 5'-ATGGCTTTTTGTGCATATCAGAT-3 '(SEQ ID NO: 6).

The PCR conditions were 32 cycles of 94 ℃ 30 seconds, 56 ℃ 30 seconds, 72 ℃ 1 minute, the PCR reaction was electrophoresed in 2% agarose gel and stained with EtBr, and then compared with the UV irradiation to compare the density and The value was analyzed by dividing by the density value.

[Experiment result]

1) Airway hyper-responsiveness (AMR) investigation

Hypersensitivity to methacholine according to the concentrations of montelukast (DR. REDDY'S) used as a positive control drug and the test substances obtained in Reference Examples 1, 2 and 7 was shown in Table 6 below.

division Penh Value According to Methacholine Dosage 0 mg / ml 10 mg / ml 20 mg / ml Normal 1.2 ± 0.1 1.7 ± 0.2 6.0 ± 2.4 Triggering group 1.5 ± 0.2 14.3 ± 3.0 29.7 ± 12.2 Preparation Example 7 (200mg / kg) 1.5 ± 0.2  7.5 ± 2.1  15 ± 4.9 Reference Example 1 (200mg / kg) 1.4 ± 0.2 8.2 ± 2.5 18.1 ± 5.9 Reference Example 2 (200mg / kg) 1.5 ± 0.2 11.1 ± 3.8 25.1 ± 4.7 Positive control group Montelukast (30mg / kg) 1.5 ± 0.2 8.4 ± 2.4 19.8 ± 5.4

As shown in Table 6, the material of Preparation Example 7 according to the present invention was shown to have a superior airway hyperresponsiveness inhibitory activity than the positive control drug. In addition, it is possible to confirm the enhanced airway hypersensitivity inhibitory activity than when the extracts of Reference Example 1 and Reference Example 2 were used alone.

2) Investigation of IgE levels in plasma

Plasma IgE values according to the concentrations of the montelukast used as the positive control agent and the test substances obtained in Reference Example 1, Reference Example 2 and Preparation Example 7 are shown in Table 7 below.

division IgE level (ng / ml) Normal 0.6 ± 0.2 Triggering group 19.7 ± 6.1 Preparation Example 7 (200mg / kg)  9.6 ± 3.2 Reference Example 1 (200mg / kg) 11.0 ± 4.8 Reference Example 2 (200mg / kg) 14.4 ± 4.6 Positive control group Montelukast (30mg / kg) 13.7 ± 5.7

As in Table 7, it can be seen that the group administered with the substance of Preparation Example 7 according to the present invention has a lower level of IgE, an inflammation mediator, than the group administered with the positive control drug. In addition, it can be confirmed that the value of IgE lower than when the extract of Reference Example 1 and Reference Example 2 were used alone.

3) Cell number in bronchial and pulmonary lavage fluid (BALF)

The neutrophils and eosinophil cell numbers in the bronchial and pulmonary lavage fluids according to the concentrations of the montelukast used as the positive control agent and the test materials obtained in Reference Example 1, Reference Example 2 and Preparation Example 7 are shown in Table 8 below.

division WBC (x10 ³ cells) NEU EOS Normal 16.3 ± 3.4 4.7 ± 2.7 Triggering group 24.0 ± 6.1 414.7 ± 168.2 Preparation Example 7 (200mg / kg)  16.3 ± 11.3 296.7 ± 300.1 Reference Example 1 (200mg / kg) 18.7 ± 7.1 369.3 ± 232.2 Reference Example 2 (200mg / kg) 17.3 ± 6.4 302.3 ± 267.9 Positive control group Montelukast (30mg / kg) 22.3 ± 7.2 466.0 ± 207.0

NEU; neutrophil (neutrophil), EOS; eosinophil (Eosinophil)

As shown in Table 8, the group administered with the substance of Preparation Example 7 according to the present invention can be seen that the number of leukocytes (neutrophils, eosinophils) associated with inflammation is lower than the group administered with the positive control drug. In addition, the extracts of Reference Example 1 and Reference Example 2 show lower numbers of neutrophils and eosinophils than those used alone.

4) Observation of lung tissue

The degree of occlusion of bronchioles and infiltration of inflammatory cells according to the concentrations of montelukast used as a positive control agent and the test substances obtained in Reference Example 1, Reference Example 2 and Preparation Example 7 are shown in Table 9 below. The stained lung tissue photographs using the H & E staining method are shown in FIG. 4. Table 9 below shows 1 = No, 2 = slight, 3 = thin, 4 = moderate, and 5 = thick depending on the degree of occlusion and inflammatory cell infiltration.

division Microscopic observation Bronchial obstruction Inflammatory cell infiltration Normal 0 ± 0 0 ± 0 Triggering group 3.3 ± 0.5 3.8 ± 0.4 Preparation Example 7 (200mg / kg) 1.8 ± 0.4 1.9 ± 0.4 Reference Example 1 (200mg / kg) 2.0 ± 0.0 2.3 ± 0.5 Reference Example 2 (200mg / kg) 2.0 ± 0.0 2.8 ± 0.4 Positive control group Montelukast (30mg / kg) 3.5 ± 0.5 3.7 ± 0.5

As shown in Table 9, the preparation of Example 7 according to the present invention was shown to have a superior bronchial blockage inhibition and inflammatory cell infiltration inhibitory activity of the lung tissue than the positive control drug. In addition, it is possible to confirm the superior bronchial blockage inhibition and inflammatory cell infiltration inhibitory activity than when the extracts of Reference Example 1 and Reference Example 2 were used alone.

5) RT-PCR

The expression rate of inflammatory mediator cytokines (IL-1β, IL-4, IL-13r2a) according to the concentration of montelukast used as a positive control drug and the test substance obtained in Preparation Example 7 is shown in Table 10 below. Table 10 below is the density of each cytokine UV irradiation divided by the actin density.

division Cytokine Expression IL-1β IL-4 IL-13r2a Normal 0.58 ± 0.27 0.58 ± 0.21 0.56 ± 0.49 Triggering group 0.94 ± 0.39 1.96 ± 0.98 1.20 ± 0.67 Preparation Example 7 (200mg / kg) 0.16 ± 0.74 0.83 ± 0.67 0.29 ± 0.96 Reference Example 1 (200mg / kg) 1.21 ± 0.62 0.14 ± 0.70 0.60 ± 0.70 Reference Example 2 (200mg / kg) 0.52 ± 0.38 0.59 ± 0.52 0.57 ± 0.39 Positive control group Montelukast (30mg / kg) 0.51 ± 0.26 1.47 ± 0.48 0.37 ± 0.31

As shown in Table 10, the group administered with the substance of Preparation Example 7 according to the present invention has a higher expression of inflammatory mediated cytokines (IL-1β, IL-4, IL-13r2a) than the group administered with the positive control drug. Appeared to be inhibited. In addition, it can be seen that the expression of inflammation mediated cytokines (IL-1β, IL-4, IL-13r2a) is suppressed than when the extracts of Reference Example 1 and Reference Example 2 were used alone.

[ Example  4: preparation of tablets]

200 mg of the mixed extract of Preparation Example 7, 10 mg of hard silicic anhydride, 2 mg of magnesium stearate, 50 mg of microcrystalline cellulose, 25 mg of starch glycolate, 113 mg of corn starch, and an appropriate amount of anhydrous ethanol were mixed in a conventional manner. Tableting.

[ Example  5: manufacture of health food]

1000mg of mixed extract (Preparation 7)

70 ㎍ Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 ㎍

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.

Example 6: Preparation of Healthy Drinks

1000 mg of mixed extract (Preparation 7)

Citric acid 1000 mg

Oligosaccharides 100 g

Plum concentrate 2 g

Taurine Soap 1 g

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 expectoration activity of Preparation Examples 1 to 11 and Reference Examples 1 and 2 shows the results measured by the phenol red method using a mouse.

Figure 2 shows the antitussive activity of Preparation Examples 1 to 11 and Reference Examples 1 and 2 shows the results of the cough suppression test using guinea pigs.

Figure 3 shows the bronchial contraction inhibitory activity of Preparation Examples 1 to 11 and Reference Examples 1 and 2 showing the relaxation rate (%) for the contraction induction agent using guinea pig extraction bronchus.

Figure 4 shows the effect of Preparation Example 7 on the bronchial obstruction and bronchial inflammatory cell infiltration inhibitory activity in lung tissue of mice sensitized and tested with OVA.

<110> Ahn-Gook Pharma Co., Ltd. <120> Pharmaceutical composition containing a herbal mixture extract <150> KR 10-2008-0055538 <151> 2008-06-13 <160> 6 <170> KopatentIn 1.71 <210> 1 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> IL-1beta forward primer <400> 1 tcatgggatg atgatgataa cctgct 26 <210> 2 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> IL-1beta reverse primer <400> 2 cccatacttt aggaagacac ggatt 25 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> IL-4 forward primer <400> 3 tcatcggcat tttgaacgag 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> IL-4 reverse primer <400> 4 gaatccaggc atcgaaaagc 20 <210> 5 <211> 22 <212> DNA <213> Artificial Sequence <220> IL-13r2a forward primer <400> 5 ggttatgcca aatgcacttg ag 22 <210> 6 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> IL-13r2a reverse primer <400> 6 atggcttttt gtgcatatca gat 23  

Claims (10)

Containing a mixed extract of ivy leaf and rhubarb as an active ingredient, wherein the mixed extract of ivy leaf and rhubarb is a mixture of ivy leaf and rhubarb extract, or an extract of a mixture of ivy leaf and rhubarb, antitussive or expectorant composition. A mixed extract of ivy leaf and rhubarb as an active ingredient, wherein the mixed extract of ivy leaf and rhubarb is a mixture of ivy leaf and rhubarb extract, or an extract of a mixture of ivy leaf and rhubarb, composition for preventing or treating respiratory diseases .  The composition according to claim 1 or 2, wherein the content ratio of the ivy leaf extract and the rhubarb extract in the mixed extract is 0.1: 1 to 10: 1 (the ivy leaf extract weight: the weight of the sulfur extract) based on the solids weight. The composition according to claim 1 or 2, wherein the content ratio of the ivy leaf extract and the rhubarb extract in the mixed extract is 0.2: 1 to 5: 1 (weight of the ivy leaf extract: weight of the yellow extract) based on the solids weight. According to claim 1 or 2, wherein the extract of the ivy leaf, the extract of the yellow leaf or the mixture of the ivy leaf and the yellow leaf is ivy leaf, the yellow leaf or the linear or branched alcohol having 1 to 4 carbon atoms in the mixture of the ivy leaf and the yellow lotus The composition is a crude extract obtained by adding one or more solvents selected from the group consisting of, or a solvent-soluble extract obtained by adding an aqueous solution of 1 to 6 linear or branched alcohols to the crude extract of the sulfur extract or the crude extract of the ivy leaf and the sulfur extract. .  The method of claim 2, wherein the respiratory disease is emphysema, bronchitis, asthma, chronic or acute bronchial contraction, wheezing infant syndrome, chronic obstructive pulmonary disease, bronchial adenomas, isolated pulmonary nodules, pulmonary tuberculosis, empyema, lung abscess, cold, flu, and lung The composition is at least one disease selected from the group consisting of histiocytosis.  7. The bronchitis of claim 6, wherein the bronchitis is selected from the group consisting of chronic bronchitis, acute bronchitis, catarrhal bronchitis, and obstructive or inflammatory bronchial disease, the asthma being bronchial asthma, atopic asthma, atopic bronchial IgE mediated asthma, non The composition is selected from the group consisting of atopic asthma, allergic asthma, and non-allergic asthma. The composition of claim 1 or 2, wherein the composition is formulated as a powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, transdermal, suppository, or sterile injectable solution. The composition of claim 1 or 2, wherein the daily dosage of the composition is 0.7 to 500 mg / kg based on the active ingredient content. Health functional food for preventing or improving respiratory disease containing the composition of claim 2.

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