KR20120060053A - Composition for the anti-inflammatory and analgesia comprising extract of propolis and Sophora japonica complex containing flavonoid - Google Patents

Abstract

PURPOSE: An anti-inflammatory analgesic composition containing propolis and Sophora japonica extract is provided to suppress acute edema and ear edema and to enhance anti-inflammation. CONSTITUTION: An anti-inflammatory analgesic composition contains propolis and Sophora japonica extract as an active ingredient. The extract contains 5.0-20.0 wt% of flavonoid as an indicator. The weight ratio of the propolis and Sophora japonica is 60 : 40-80 : 20. A functional food contains the propolis and Sophora japonica extract as an active ingredient. A method for preparing the propolis and Sophora japonica extract containing flavonoid comprises: a step of extracting, cooling, and filtering propolis and Sophora japonica to prepare liquid; a step of adding sodium hydrogen carbonate to the liquid and aggregating at -20°C to -30°C; a step of removing wax and collecting filtered liquid; a step of maturing the filtered liquid at room temperature for 1-7 days; a step of centrifuging the liquid at 3,000-5,000 rpm, reducing pressure, pulverizing, and sterilizing.

Description

TECHNICAL FIELD The present invention relates to a flavonoid-containing anti-inflammatory analgesic composition comprising propolis and an extract of Isofura as an active ingredient.

The present invention relates to an anti-inflammatory analgesic composition and a method for preparing the same, and more particularly, to an extract of propolis and Sophora japonica as an active ingredient, wherein the extract comprises 5.0 to 20.0% Of flavonoids and a method for preparing the extracts.

Inflammation refers to the pathological condition of an abscess formed by the invasion of external bacteria. The inflammation reaction is a biological defense reaction process for repairing and regenerating the damage caused by invasion that causes any change in the biological cells or tissues. Inflammatory mediators and immune cells are involved in local blood vessels and body fluids when they are infected with damage to cells (cells) or external infectious agents (bacteria, fungi, viruses, various allergens) A series of complex physiological responses including invasion, cell migration, and tissue destruction, and external symptoms such as erythema, swelling fever, and the like. In normal cases, the inflammatory reaction removes the external infectious agent and regenerates the damaged tissue to regenerate the organism's function. However, if the antigen is not removed or the internal substance causes the inflammatory reaction to occur excessively or continuously, the major pathological condition of the disease (irritable disease, Chronic inflammation), and become obstacles in the treatment process such as transfusion, drug administration, or organ transplantation.

Nitric oxide (NO), an inflammatory factor involved in inflammatory reactions, is activated by nitric oxide synthase (NOS) from L-arginine via the intermediate metabolite hydroxyarginine, And citrulline are made. NO is a biologically active substance with a very small molecular weight and acts on the vascular system in vivo to act as a mediator involved in vasodilation, platelet adhesion and aggregation, neurotransmission, and digestive organ exercise. (Neurotransmission, blood coagulation, blood pressure control, immunity against cancer cells, etc.), and it is produced not only in inflammatory cells but also in non-immune cells, and acts to protect against microbial infection. In general, NO is present in the form of gas in the atmospheric state. It is not only poisonous because it is free radical structure, it is very unstable with a half-life of 6 to 10 seconds, and stabilized NO ₃ (nitrate) and NO ₂ (nitrite) (Snyder, SH et al., Scientific American, May, pp28-35, 1992).

On the other hand, NOS participating in NO production is classified as a calcium (Ca ^{2 +)} or calmodulin (camodulin), there are two main i.e., cNOS (constitutive NOS) and iNOS (inducible NOS) in accordance with the dependence whether, cNOS is calcium, INOS is present in macrophages, hepatocytes, and cancer cells, and IL-1β (interlukin-1β) is present in brain, epithelial cells, neutrophils, and gastric mucosal cells, and iNOS is independent of calcium or calmodulin. , Cytokines such as IFN-γ (interferon-γ), TNF-α (tumor necrosis factor-α), and bacterial LPS (lipopolysaccharide) endotoxins (Dinerman, JL, et al., Circ Res., 73, pp217-222, 1993). The expression of COX-2 is closely related to the expression of COX-2 (cyclooxygenase-2) by activating COX-2 (prostaglandines) (Daniel S., et al., Proc. Natl. Acad. Sci. USA, 90, pp. 7240- 7244, 1993).

Therefore, various efforts have been made to secure a substance that inhibits the production of NO by iNOS and develop it as a pharmaceutical product.

Inflammatory diseases caused by these inflammatory reactions include gastric inflammation, colitis, arthritis, nephritis, hepatitis, cancer or degenerative diseases (Gobert AP et al., J. Immunol. 168 (12), pp6002-6006, 2002; Dijkstra G. et al., Scand. J. Gastroenterol. 37 (5), pp 546-554, 2002, Sakac V. and Sakac M. Med. Pregl 53, pp 463-474, 2000; Albrecht EW et al., Am. Sartor L. et al., Biochemical Pharmacol. 64 (11), pp. 1465-1474, 2002; et al., Carcinogenesis, 23 (6), pp. 229-237, 2002; Sadowska-Krowicka H. et al., Proc. Soc. , pp. 983-991, 2002).

It is also called Painting on the Basilia Gorge, killing the roundworms, and controlling the openings, furrows, and storm winds. Painted tree is a deciduous tree with beans and 10 m tall. Leaves are composed of 6 ~ 8 pairs of foliate leaves, and are planted in various places throughout the country. It has been presumed as a herbal medicinal preparation in the preliminary herbal medicines such as Donguibogam, Hyangbokgungbang, and Gwangseongwa and related literature, but these prescriptions are based on the simple method of differentiating the external shape of the medulla and the manufacturing method of herbal medicine and the solution Recently, a patent having an active ingredient of a routine, which is one type of flavonoid, has been disclosed in an anti-inflammatory analgesic composition containing an extract of Lycopene (Lycopene) as an active ingredient in Korean Patent Laid-Open No. 2008-0101352.

In recent years, the treatment of inflammatory periodontal disease of 100% methanol extracts from 100% methanol has been disclosed (Lee et al., Journal of Korean Periodontology, 35 (1): 9-17, 2005), and Korean Patent No. 509,682 Korean Patent No. 539,457 discloses a composition for the prevention and treatment of hyperlipemia, atherosclerosis and fatty liver comprising an extract of Chrysanthemum morifolium as disclosed in Korean Patent No. 539,456 Korean Patent Laid-Open No. 2006-0033070 discloses a composition for prevention and treatment of cancer including extracts of Chrysanthemum morifolium, in which various herbal medicines, including bean sprouts, are used for a bathing solution for the treatment of hemorrhoids or gynecological diseases, Lt; / RTI > In a mouse model experiment in which cerebral infarction was induced in a foreign country, the extracts of the microorganisms such as Ginkgo biloba have the effect of reducing the production of interleukin-1, microglial cell activity and brain cell death, (Lao CJ et al., Am J Chin Med. 33 (3): 425-438, 2005).

Propolis is a kind of protector that protects the beehive by blocking the tearing gap of the honeycomb to prevent the entry of rainwater or wind, by preventing the invasion of bacteria or virus from the outside by the honey bee at the entrance of the beehive. In addition, propolis disinfects and disinfects inside the honeycomb, keeping the inside of the honeycomb in aseptic condition and allowing the honeycomb inside the honeycomb to grow safely. The origins of propolis originated in Greece, pro means greek before and polis means city. Therefore, the combination of the two etymologies means 'the front of the city'. Here 'city' means honeycomb, which means 'material that prevents safety and disease in front of the honeycomb'. According to the Dong-bo-gyung, propolis is recorded under the name of "房 房", which is effective in relieving asthma. In the modern world, cold, allergy, atopy, asthma, burns, It is widely used as a private medicine for various immune diseases. Such propolis is considered to be useful as a natural antibiotic food which can be used as a substitute for antibiotic in the modern age in which resistance is induced by the use of various antibiotics and super virus is generated, thereby contributing to promotion of public health.

However, there has been no report on the inhibition of inflammatory mediators and cells by some of the components of propolis and extracts, and the characteristics of natural products such as propolis and sorghum are different depending on the content of indicator components There are various problems such as difficulty in standardization of the extract, difficulty in commercialization because it is difficult to reproducibly obtain an active fraction having excellent activity such as anti-inflammatory analgesic, blood circulation improvement, arthritis treatment and the like.

Accordingly, the present inventors have studied and found that the anti-inflammatory effect of propolis and the extract of Lycopersiconum majorifolia according to the specific active ingredient having a remarkable antiinflammatory effect and the content thereof in the propolis and Lycopersicon esculentum extracts is as follows: flavonoid 5.0-20.0 weight % Of the propolis extract and the extract of Lycopersicon esculentum have an excellent anti-inflammatory effect.

As a result, the main object of the present invention is to provide an anti-inflammatory analgesic composition comprising a propolis and flavonoid extract containing flavonoid as an active ingredient.

It is still another object of the present invention to provide a process for producing propolis and an ingot extract containing the flavonoid having an excellent anti-inflammatory effect.

In order to achieve the above object, the present invention pro containing flavonoids of 5.0 ~ 20.0% by weight as an indicator substance u (Propolis) and goehwa (Sophora japonica ) extract as an active ingredient.

Further, the present invention provides a method for producing an extract, comprising the steps of: a) extracting, cooling and filtering a 2-4-fold amount of a water, alcohol or alcohol aqueous solution at a weight ratio of propolis and bean to 60:40 to 80:20 to obtain a filtrate;

b) adding sodium hydrogencarbonate to the filtrate to aggregate the waxy components of propolis at -20 ° C to -30 ° C and then filtering to obtain a filtrate from which wax is removed;

c) aging the filtrate for 1 to 7 days at room temperature to obtain a stock solution; And

d) centrifuging the stock solution at 3,000 ~ 5,000 rpm, and concentrating the filtrate under reduced pressure, freeze-drying, pulverizing and sterilizing the filtrate to obtain a propolis and a sprout extract

The present invention provides a flavonoid-containing propolis extract and a method for producing the same.

The propolis and ingrown extracts containing the flavonoids of the present invention are substances derived from natural materials and have no toxicity and side effects and have excellent antiinflammatory effects such as anti-inflammatory action, acute edema-inhibiting action and acute arthritis-suppressing action, Can be effectively used as safe and effective medicines and individually recognized health functional foods for the treatment and prevention of inflammatory diseases such as arthritis, otitis media, edema, pain or degenerative diseases caused by inflammation and inflammation reaction.

FIG. 1 is a graph showing the UV spectrophotometer results of propolis and extracts containing flavonoids according to the present invention (Standard Curve is shown and the correlation coefficient is R ² = 0.99677).

To achieve the above object, the present invention provides an extract of Propolis and Sophora japonica as an active ingredient, wherein the extract contains 5.0-20.0% by weight of flavonoid as an indicator substance An anti-inflammatory analgesic composition.

In the present invention, the flavonoid is characterized by having 5.0 to 20.0% by weight, and exhibits excellent anti-inflammatory effect against arthritis, otitis media, edema, pain or degenerative diseases caused by inflammation and inflammatory reaction have.

Further, the present invention provides a method for producing an extract, comprising the steps of: a) extracting, cooling and filtering a 2-4-fold amount of a water, alcohol or alcohol aqueous solution at a weight ratio of propolis and bean to 60:40 to 80:20 to obtain a filtrate; b) adding sodium hydrogencarbonate to the filtrate to aggregate the waxy components of propolis at -20 ° C to -30 ° C and then filtering to obtain a filtrate from which wax is removed; c) aging the filtrate for 1 to 7 days at room temperature to obtain a stock solution; And d) centrifuging the stock solution at 3,000 ~ 5,000 rpm and concentrating the filtrate under reduced pressure, freeze-drying, crushing and sterilizing the filtrate to obtain a propolis and a sprout extract. And a method for producing an extract from a barb. In the present invention, the flavonoid may be in an amount of 5.0 to 20.0% by weight.

In the present invention, the propolis extract may be a powder, and the alcohol may be a lower alcohol having 1 to 6 carbon atoms such as methanol, ethanol, propanol, butanol, and the like.

Hereinafter, the present invention will be described in detail.

The present invention relates to a process for preparing a propolis extract and a crude extract containing a certain amount of flavonoid as an active ingredient, and to an anti-inflammatory analgesic composition comprising the flavonoid as an active ingredient.

The antiinflammatory analgesic composition of the present invention contains propolis and an extract of Lycopersicum as an active ingredient, and the propolis and the Lycopene extract comprise 5.0 to 20.0% by weight of flavonoid as indicator material, more preferably 5.0 to 20.0% 15.0% by weight. It is preferred that the propolis and bean curd are contained in a weight ratio of 60:40 to 80:20. However, since the content of effective physiologically active substances contained in natural products such as propolis and bean sprouts may vary greatly depending on the place of production, the collection time, storage period, and storage condition, It is more preferable to limit the content of the indicator material.

Therefore, in the present invention, flavonoids are selected as indicator substances for obtaining propolis and extracts which maximize the expression of the desired anti-inflammatory effect.

The propolis and the extract of L. sp. Used in the treatment of the inflammatory diseases of the present invention can be selected by extracting propolis and flavonoids which are specific indices from the extract itself (herbal medicine extract) extracted from the sprouts, or the propolis and L. extract. At this time, the blending ratio is preferably determined by the content of flavonoid, which is an indicator substance and an active ingredient, rather than the original content of propolis and sorghum, preferably 5.0 to 20.0% by weight of flavonoid in the propolis and the sprout extract, Preferably from 5.0 to 15.0% by weight of flavonoid.

In the present invention, when the content of the flavonoid is less than 5.0% by weight, it exhibits a relatively low effect on the treatment of arthritis and the like. Therefore, it is preferable that the content of the flavonoid is 5.0% by weight or more. In addition, the upper limit of the flavonoid content in the present invention is not particularly limited. However, if the content exceeds a certain level, the effect is not further increased, and it is not preferable from the technical and economical point of view. By weight or less, more preferably about 15.0% by weight or less.

In the present invention, when the indicator substance is contained in an amount of 5.0-20.0 wt% of flavonoid, which is an active substance, it exhibits a synergistic effect of the desired drug efficacy and thus can exhibit an antiinflammatory effect of the propolis and the extract of the ingot, which is more excellent. It is impossible to exclude the action of other components other than the above-mentioned flavonoid among the obtained propolis and extracts.

A process for preparing propolis and an extract of Isoflavones containing 5.0-20.0 wt% of flavonoids having a significant anti-inflammatory effect according to the present invention comprises the following steps.

a) extracting with 2 to 4 times water, alcohol or alcohol aqueous solution, cooling and filtering to obtain a filtrate with a weight ratio of propolis and bean to 60:40 to 80:20;

b) adding sodium hydrogencarbonate to the filtrate to aggregate the waxy components of propolis at -20 ° C to -30 ° C and then filtering to obtain a filtrate from which wax is removed;

c) aging the filtrate for 1 to 7 days at room temperature to obtain a stock solution; And

d) centrifuging the stock solution at 3,000 ~ 5,000 rpm, and then concentrating the filtrate under reduced pressure, freeze-drying, crushing and sterilizing the filtrate to obtain a propolis and a sprout extract.

The preparation method is described in more detail. In step (a), 2 to 4 times of a water, alcohol or alcohol aqueous solution is added to the propolis and the original raw material of a weight ratio of 60:40 to 80:20 in a weight ratio of 12 to 36 hours, It is extracted 5 times repeatedly, then slowly cooled at room temperature, and then separated by filtration by centrifugation or the like. Water, an alcohol or an alcohol aqueous solution of 2 to 4 times the weight of the mixed preparation is added to the residue, and the mixture is warmed to re-extract and then filtered, mixed with the previous filtrate, and filtered. In the present invention, the extraction efficiency can be increased by re-extracting and filtering by adding water or an aqueous alcohol solution to the residue as described above. When the amount of water, alcohol or alcohol aqueous solution used for extraction is less than 2 times, the solubility of the extract is deteriorated The extraction efficiency is inferior. When the use amount exceeds 4 times, the use amount of the aqueous alcohol solution used for layer separation increases, and economical such as long time for decompression and concentration may take place, or there may be problems in handling.

In the present invention, the alcohols may be aliphatic or aromatic alcohols commonly used in the field, and preferably aliphatic alcohols, more preferably lower alcohols having 1 to 6 carbon atoms such as methanol, ethanol, propanol, It is preferable to use alcohol. In the case of the alcohol aqueous solution, it is preferable to use an aqueous 60 to 80% ethanol solution.

In the present invention, a method of re-extraction after primary extraction has been adopted. In the case of mass production of herbal medicine extracts, since the water content of the herbal medicine itself is high even if the filtration is effected effectively, the extraction efficiency is lowered only by the primary extraction alone This is to prevent this. In addition, it was found that 80 ~ 90% of the total extraction amount was extracted by the second extraction, and it is considered that the extraction of the third and more stages is not economical.

As described above, the extract obtained by extracting with water, alcohol or alcohol aqueous solution is filtered and concentrated, and unnecessary proteins, polysaccharides and fatty acids contained in the filtrate are purified. In the present invention, In an amount of 2 to 4 times the amount of the alcohol aqueous solution as a solvent to obtain a solvent fraction, whereby the impurities can be purified. The alcohol used for the layer separation is preferably a lower alcohol having 1 to 6 carbon atoms such as methanol, ethanol, propanol, butanol and the like, more preferably 60 to 80% ethanol aqueous solution. When the amount of the lower alcohol aqueous solution used is less than that of the filtrate in the layer separation, fine particles are formed by unnecessary components such as fatty acids, so that the layer separation is not smooth and the extractable content of the active active material is lowered It is not efficient.

In step b), 0.1 to 0.5% by weight, more preferably 0.1 to 0.3% by weight, based on the filtrate, of sodium hydrogen carbonate as an edible acidity regulator is added to the filtrate to increase the acidity to induce color development . If the amount of sodium hydrogencarbonate is less than 0.1% by weight, color development is not performed well. If it exceeds 0.5% by weight, transparency disappeared to such an extent as to be discolored. It is then stored at -20 ° C to -30 ° C in an ultra-low temperature refrigerator or the like to coagulate the beeswax component of propolis. When the temperature is higher than -20 ° C, the wax is reused. When the temperature is lower than -30 ° C, the raw material itself is freezing. Therefore, the propylene wax It is preferable that the components are agglomerated and filtered with a broad wood to remove wax.

In the step c), the filtrate is aged at room temperature for 1 to 7 days to obtain a stock solution. The filtrate is aged at room temperature for 1 to 7 days to induce fermentation of the flavonoid, which is an indicator substance. In the course of aging, the color changes to dark brown and has a unique fragrance. Generally, it is not aged well for 1 day or less, and when aged for 7 days or more, it is economically disadvantageous and it is inefficient in mass production.

In step d), the filtrate obtained by centrifuging the stock solution at 3,000 ~ 5,000 rpm is subjected to vacuum concentration, freeze-drying, pulverization and sterilization to obtain a propolis and an extract.

In the present invention, when the decompression concentration temperature is less than 60 ° C, it is difficult to remove 100% of the solvent. When the decompression concentration is higher than 80 ° C, a problem may arise in the stability of the extract. When the rpm is less than 3,000, Is difficult to separate, and if it is 5,000 rpm or more, the stability of the extract may be problematic.

The concentrate was vacuum-dried at 60 to 80 ° C at 0.08 to 0.3 pa and then pulverized and sterilized at 30 to 80 mesh to obtain a powdery propolis extract and an autoclaved extract containing 5.0 to 20.0 wt% of flavonoid as an active ingredient can do.

Using the ICR mouse and male SD regret (supplied from Daejeon Chemical Research Institute, Korea) for the propolis and the raw extracts containing a certain amount of the flavonoids obtained as described above, the test for inhibition of edema and the exfoliation of flavonoid-containing propolis and extracts Inhibition test, and arthritis treatment.

As a result, it was confirmed that the propolis extract of the present invention has an excellent anti-inflammatory effect such as anti-inflammatory action, acute edema-inhibiting action, and acute arthritis-inhibiting action.

Therefore, it can be seen that the propolis and the extract of the present invention are safe substances having excellent anti-inflammatory effect and no toxicity.

Accordingly, the propolis and the compacted extract containing 5.0 to 20.0% by weight of flavonoid as an active ingredient of the present invention can be effectively used for medicines and functional foods for the treatment and prevention of inflammatory diseases, and are not limited thereto.

When the composition of the present invention is used as a medicine, the natural extract alone may be included alone, but it may further include appropriate carriers, excipients and diluents conventionally used in the production of pharmaceutical compositions, For example, powders, tablets, granules, capsules or injections, and the like.

Examples of the carrier or excipient include water, dextrin, calcium carbonate, lactose, propylene glycol, liquid, paraffin, physiological saline, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gelatin, calcium phosphate, calcium There may be used one or more of them, but it is not limited to these, and examples thereof include, but are not limited to, polyvinylpyrrolidone, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, Of the carrier and excipient are all usable. In addition, when the antioxidant composition is weakly formulated, it may further contain conventional fillers, extenders, binders, disintegrators, surfactants, anti-coagulants, lubricants, wetting agents, perfumes, emulsifiers or preservatives.

When the propolis and the lycopene extract according to the present invention are formulated into tablets by a conventional method, it is preferable that the lecithin, microcrystalline cellulose, magnesium stearate, etc., which is used as a base, and the lycopene extract are used in a ratio of 1: 1 desirable.

In addition, the propolis and the extract of the present invention can be used for edible and medicinal purposes, and there is no particular limitation on the dosage of the propolis and the extract of the present invention, and the degree of absorption, body weight, age, sex, health condition, Method, excretion rate, severity of disease, and the like. However, in order to obtain the desired effect, the propolis and the extract of the present invention may be used in an amount of 0.001 to 50 mg per kg of body weight per day, 10 mg per day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

The propolis and extracts of the present invention can be administered to mammals such as rats, mice, and livestock in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.

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

Example  1: propolis containing flavonoids and Cow  Preparation of extract

After washing with water and drying well, propolis and bean sprouts (70: 30 weight ratio) were added to three times 70% ethanol aqueous solution and extracted with hot water twice with stirring for 24 hours. The hot-water extract was slowly cooled to room temperature, then centrifuged to remove residues, and the filtrate was combined. Sodium bicarbonate was added to the filtrate in an amount of 0.1 wt% of the filtrate, and the beeswax component of the propolis was agglomerated at -20 ° C, followed by filtering with wax to remove the beeswax. The filtrate was aged at room temperature for 5 days to obtain a stock solution. The stock solution was centrifuged at 3,000 rpm, concentrated under reduced pressure, vacuum dried and freeze-dried at 60 ° C under 0.08 pa, and then pulverized and sterilized in 80 mesh To obtain a propolis and an ingot extract. The content of flavonoid in the propolis and the extract of the lycopersicum extracted and purified by the above-mentioned method was 15.0% by weight.

In addition, the flavonoid-containing propolis and the compacted extract obtained by the above-mentioned method were analyzed by UV under the following conditions, and the results are shown in FIG.

[UV analysis method]

1) Standard substance: Quercetin is made to 1 mg / ml and diluted to 100 and 20.10 times to prepare a calibration curve.

2) General reagents: Ethanol

3) General test solution: 10% aluminum nitrate solution, 1 M potassium acetate solution, 80% ethanol, 90% ethanol

4) Preparation of propolis and extracts for UV analysis:

   4.1) After vacuum drying and lyophilization, pulverize and sterilize to 80 mesh, and weigh 100 mg of propolis and lycopersicum extract and dissolve with 20 mg of 90% ethanol.

   4.2) Centrifuge (3000 rpm, 10 min) to take supernatant.

   4.3) Extract the residue three times with 8 ml of 80% ethanol.

   4.4) Combine total extracts and make up to 50 ml with 80% ethanol.

5) Test operation:

   5.1) Take 0.5 ml of the standard solution and the test solution into the two test tubes for the reference solution and the test solution.

   5.2) Add 1.5 ml of ethanol, 0.1 ml of 10% aluminum nitrate solution, 0.1 ml of 1 M potassium acetate solution and 2.8 ml of distilled water and stir well. A blank test of standard substances and samples for each concentration is carried out by adding 0.1 ml of distilled water instead of 10% aluminum nitrate.

   5.3) After standing at room temperature for 40 minutes, measure the absorbance of the liquid layer at 415 nm.

As shown in Fig. 1, when the correlation coefficient is R ² = 0.99677, respectively.

Experimental Example  1: propolis and Cow  extract TPA  Induction ear edema TPA - induced mouse ear edema ) Suppression test

In order to confirm whether the anti-inflammatory effect of the propolis and the lycopersicum extract containing the flavonoid of the present invention is applicable to animal experiments, the following experiment was conducted.

TPA (2.5 ㎍ / 20 ㎕) dissolved in acetone was dissolved in acetone for 1 hour after oral administration of propolis and extract extracts 10, 100, 200 and 400 ㎎ / Was used to induce edema in the ear of an experimental animal. At this time, 1 mg / kg of indomethacin was used as a conventional anti-inflammatory agent for comparison, and propolis had a content of 5.0 mg / kg of flavonoid and 5.0 mg / kg of flavonoid and 400 mg / kg of rutin content of 20.0% Respectively. Four hours after induction of edema, ear edema was measured in each experimental group. At the time of measurement, the cervical dislocation method was used to precisely measure the mouse after culling. The results are shown in Table 1 below.

TPA-Induced Inhibition of Ear Edema in Propolis and Wheat Extracts Test group Mouse ear thickness (mm) Inflammation inhibition rate (%) Control group 0.75 + - 0.01 - Propolis and sorghum
(400 mg / kg-flavonoid content 15.0% by weight) 0.44 + 0.02 41% Propolis and sorghum
(200 mg / kg-flavonoid content 15.0% by weight) 0.60 + 0.02 20% Propolis and sorghum
(100 mg / kg-flavonoid content 15.0% by weight) 0.66 ± 0.01 12% Propolis and sorghum
(10 mg / kg-flavonoid content 15.0% by weight) 0.71 + - 0.03 5% Propolis
(400 mg / kg-flavonoid content 5.0% by weight) 0.65 + 0.03 13% (400 mg / kg-rou- tine content 20.0% by weight) 0.48 ± 0.01 36% Indomethacin (1 mg / kg) 0.41 + 0.02 45% Data represent the mean of difference in ear thickness (mm) ± S.E.M (n = 12)

As shown in the above Table 1, the inhibition rate of inflammation was the highest at 400 mg / kg of the extract of propolis and the extract of Iris, which was 41%, considering the data deviation compared with the inflammation inhibition rate of 45% of the existing inflammatory drug indomethacin Indicating an equivalent level of efficacy. Propolis was used at a dose of 400 mg / kg, but propolis was inhibited by 13%, and the ingestion was inhibited by 36%. However, the combination of the compound extracted in Example 1 and the anti- , And it was confirmed that the synergy effect of propolis and the extract of Lycopersicon esculentum appeared. Therefore, it was confirmed that the propolis and the extract of the present invention inhibit TPA-induced ear edema in a concentration-dependent manner.

Experimental Example  2: Propolis and Cow  By extract Base agent Arachidonic acid  Induction ear edema Arachidonic acid -induced mouse ear edema ) Suppression test

In order to confirm whether or not the anti-inflammatory effect of the propolis and the lycopersicum extract containing the flavonoid of the present invention is applied to animal experiments, the following experiment was conducted.

Seven weeks old ICR mice were divided into each experimental group and 2 ㎍ / 20 ㎕ of arachidonic acid dissolved in acetone was dissolved 1 hour after oral administration of propolis and extract extracts 10, 100, 200 and 400 ㎎ / A 29G needle was used to induce edema in the ear of the experimental animal. At this time, 1 mg / kg of indomethacin was used as a conventional anti-inflammatory agent for comparison, propolis was contained in a concentration of 5.0 mg / kg of flavonoid, 5.0 mg / kg of flavonoid, 400 mg / kg of rutin, Were used. One hour after induction of edema, ear edema was measured in each experimental group. At the time of measurement, cervical dislocation was used to obtain accurate measurement after culling of the experimental mice. The results are shown in Table 2 below.

Effect of Arachidonic Acid Induced by Propolis and Wheat Extract on Ear Edema Test group Mouse ear thickness (mm) Inflammation inhibition rate (%) Control group 0.69 ± 0.02 - Propolis and sorghum
(400 mg / kg-flavonoid content 15.0% by weight) 0.37 + 0.02 46% Propolis and sorghum
(200 mg / kg-flavonoid content 15.0% by weight) 0.49 + 0.02 29% Propolis and sorghum
(100 mg / kg-flavonoid content 15.0% by weight) 0.60 ± 0.01 13% Propolis and sorghum
(10 mg / kg-flavonoid content 15.0% by weight) 0.68 ± 0.03 0 % Propolis
(400 mg / kg-flavonoid content 5.0% by weight) 0.61 + 0.03 12% (400 mg / kg-rou- tine content 20.0% by weight) 0.38 + 0.02 45% Indomethacin (1 mg / kg) 0.35 + 0.04 49% Data represent the mean of difference in ear thickness (mm) ± S.E.M (n = 12)

As shown in the above Table 2, the inhibition rate of inflammation was the highest at the concentration of 400 mg / kg of propolis and the extract of Isoflavin as 46%, which is equivalent to that of the inflammation inhibition rate of 49% of the existing inflammatory drug indomethacin Respectively. In the case of propolis or bean sprouts used alone, the rate of inhibition of propolis and propolis was inhibited by 12% and 45%, respectively, when 400 mg / kg was used. It was confirmed from the animal experiment that the combination agent extracted in Example 1 had a superior inflammation inhibitory effect, and it was confirmed that the synergy effect of the propolis and the extract of the lycopene was exhibited. Therefore, it was confirmed that the propolis extract and the extract of the present invention inhibit arachidonic acid-induced ear edema in a concentration-dependent manner.

Experimental Example  3: propolis and Cow  Induction Stretching of Acetic Acid with Extract Acetic acid - induced  writhing response in mice ) Suppression test

In order to confirm whether or not the propolis and the extract of the present invention were effective in alleviating and suppressing not only the edema of the ear but also the pain relief of the ear, the ICR male mice fasting for 12 hours were administered propolis and the test material of 10, 100, 200 , 400 mg / kg were orally administered. Diclofenac 25 mg / kg was used as the reference drug, propolis was used in an amount of 5.0 mg / kg of flavonoid and 5.0 mg / kg of flavonoid in a single dose. After 30 minutes of oral administration of the drug, 0.7% acetic acid-saline solution (0.1 mg / 10 g) was intraperitoneally injected, and the number of writhing occurred for 10 minutes after 10 minutes of injection, 1959), and the results are shown in Table 3 below.

Inhibitory Effect of Acetic Acid Induced Stretching by Propolis and Wheat Extract Test group Torsion frequency
(Writhing number) Pain inhibition (%) Control group 29.12 + - 1.23 - Propolis and sorghum
(400 mg / kg-flavonoid content 15.0% by weight) 18.21 + - 1.01 37% Propolis and sorghum
(200 mg / kg-flavonoid content 15.0% by weight) 22.19 ± 1.65 24% Propolis and sorghum
(100 mg / kg-flavonoid content 15.0% by weight) 23.12 ± 1.72 21% Propolis and sorghum
(10 mg / kg-flavonoid content 15.0% by weight) 28.26 ± 1.49 3% Propolis
(400 mg / kg-flavonoid content 5.0% by weight) 20.21 + - 1.01 31% (400 mg / kg-rou- tine content 20.0% by weight) 20.04 ± 1.11 31% Diclofenac (25 mg / kg) 14.71 ± 1.28 49% Data represent the mean of difference in ear thickness (mm) ± S.E.M (n = 12)

As shown in the above Table 3, when the propolis and the extract of Lycopersicon esculentum were 400 mg / kg, the inhibition rate of pain was the highest at 37%, which is somewhat weaker than that of the conventional pain reliever Diclofenac, which is 49% The non-steroidal anti-inflammatory analgesics such as NSAIDs may induce cardiovascular diseases upon long-term administration, thus confirming that the natural extract of Example 1 is an alternative. In addition, propolis was used at a dose of 400 mg / kg, and propolis was inhibited by 31%, and lysozyme was inhibited by 31%. It was confirmed that the combination agent extracted in Example 1 had a better pain suppressing effect than that of the complex agent extracted in Example 1, and synergistic effects of propolis and the extract of the lycopene were shown. It can be confirmed from animal experiments that the compound has an analgesic effect and that the propolis and the extract of the present invention have an analgesic effect on acetic acid-induced analgesia in a concentration-dependent manner

Experimental Example  4: Propolis and Cow  Test of inhibitory effect of extract on central nervous system pain hot plate test )

10, 100, 200, and 400 mg / kg of the test substance propolis and the seedlings were orally administered to the ICR male mice that were fasted for 12 hours before the experiment. Diclofenac 25 mg / kg was used as the reference drug, propolis was used in an amount of 5.0 mg / kg of flavonoid and 5.0 mg / kg of flavonoid in a single dose. The pain response was carefully placed on a hot plate (Ugo Basile, Italy) maintained at 55 ° C and the time from contact with the hot plate to licking or jumping back (Woolfe and Mac Donald, 1944). The results are shown in Table 4 below.

Inhibition of central nervous system pain by propolis and extracts Test group Reaction time (Sec) Pain inhibition (%) Control group 15.13 + - 1.76 - Propolis and sorghum
(400 mg / kg-flavonoid content 15.0% by weight) 22.08 ± 1.09 31% Propolis and sorghum
(200 mg / kg-flavonoid content 15.0% by weight) 20.21 1.51 22% Propolis and sorghum
(100 mg / kg-flavonoid content 15.0% by weight) 19.32 ± 1.62 22% Propolis and sorghum
(10 mg / kg-flavonoid content 15.0% by weight) 15.54 ± 1.09 3% Propolis
(400 mg / kg-flavonoid content 5.0% by weight) 20.19 ± 2.07 25% (400 mg / kg-rou- tine content 20.0% by weight) 19.11 + 1.95 21% Diclofenac (25 mg / kg) 24.22 + 1.55 38% Data represent the mean of difference in ear thickness (mm) ± S.E.M (n = 12)

As shown in Table 4, the inhibition rate of pain was the highest at a concentration of 400 mg / kg of the propolis extract and the extract of Lycopersicon esculentum at 31%, which is somewhat weaker than the 38% inhibitory effect of diclofenac, The steroidal anti-inflammatory analgesic agent can induce cardiovascular diseases upon long-term administration, thus confirming that the natural extract of Example 1 is an alternative. In addition, propolis was used at a dose of 400 mg / kg, and propolis was inhibited by 25%, and lysine by 21%. It was confirmed that the combination agent extracted in Example 1 had a better pain suppressing effect than that of the complex agent extracted in Example 1, and synergistic effects of propolis and the extract of the lycopene were shown. In particular, it was confirmed that drug efficacy increased dose-dependently by obtaining dose-dependent results.

Experimental Example  5: Propolis and Cow  Effectiveness of treatment with acute arthritis by extract (λ Carrageenan induced  hind paw edema )

In order to test the therapeutic effect of the propolis and the extract of the present invention on acute arthritis, the following experiment was conducted. Diclofenac 25 mg / kg (used as a comparative drug), propolis and extracts 10, 100, 200 and 400 mg / kg were orally administered to male SD regrets (about 200 g) orally, and 1% 100 쨉 l of physiological saline was injected into the left foot part to induce acute foot edema. Edema was measured using a plethysmometer (LE 7500) after 3 hours and the results are shown in Table 5 below.

Effect of Propolis and Wheat Extract on the Treatment of Acute Arthritis
Test group % Growth of the foot species (mean ± s.e.m.) 1 hours 2 hours 3 hours 4 hours
Control group
29.04 ± 2.10
59.87 ± 2.45
74.25 + 2.03
75.62 ± 2.13
10
30.89 ± 1.98
61.11 + - 2.22
72.65 ± 2.69
72.12 + - 2.44
100
29.62 + - 3.05
61.09 + - 2.44
68.51 + - 2.45 *
64.29 1.51 *
200
31.09 ± 2.11
49.99 ± 2.14
58.03 ± 2.14 **
58.01 + - 2.01 **
400
29.22 + - 2.82
49.35 ± 2.37 *
45.18 ± 2.93 **
44.23 + - 2.69 **
Diclofenac
(25 mg / kg)
27.83 + - 2.57
37.98 + 2.85 *
41.18 + - 2.42 **
35.23 + - 2.61 **

As shown in the above Table 5, the treatment effect of acute arthritis was the highest when the concentration of 400 ㎎ / ㎏ was 4 hours after the propolis and the ingestion of the extract, and the growth rate of the foot was 44.23 ± 2.69. It was confirmed that the propolis and the compacted extract of the present invention had a remarkable effect on the treatment of acute arthritis in a concentration-dependent manner, although there was a slight difference of 35.23 ± 2.61 compared to the analgesic agent Diclofenac (Diclofenac).

As described above, it has been confirmed through an animal experiment that the propolis and the compacted extract containing the flavonoid of the present invention have a strong anti-inflammatory and analgesic effect as well as an acute arthritis-inhibiting effect.

Hereinafter, formulation examples of the pharmaceutical composition will be described, but the present invention is not intended to be limited thereto but is specifically described.

Manufacturing example  1: Preparation of tablets

The tablets for oral administration are prepared by the wet granulation method and the dry granulation method using the following composition with the propolis and the extract of Isofura containing the flavonoid of the present invention.

[Furtherance]

Propolis and seed extract ............... 200 mg

Light anhydrous silicic acid ............................ 10 mg

Magnesium stearate ................... 2 mg

Microcrystalline cellulose 50 mg

Sodium starch glycolate ..................... 25 mg

Lactose .................................... 101 mg

Povidone ................................... 12 mg

Anhydrous ethanol ................................

Manufacturing example  2: Manufacture of ointment

An ointment preparation is prepared with the following composition using a propolis and an ingot extract containing a flavonoid of the present invention.

[Furtherance]

Propolis and sprouts extract .......... ,,,,,,,. 5 g

Cetyl palmitate ............................ 20 g

Ethanol .................................... 40 g

Stearyl alcohol ............................ 40 g

Myristic isopropyl ............................ 80 g

Sorbitan monostearate ................... 20 g

Polysorbate .............................. 60 g

P-hydroxybenzoic acid profile .................... 1 g

Methyl paraoxybenzoate ..................... 1 g

Phosphoric acid and purified water ............................

Manufacturing example  3: Preparation of injection

The propolis and flavonoid extract containing the flavonoid of the present invention are used to prepare injections of the following composition.

[Furtherance]

Propolis and Bulgur extract .............. 100 mg

Mannitol ................................. 180 mg

Sodium Hydrogen Phosphate ........................ 25 mg

Mainly used purified water ........................ 2,974 mg

Manufacturing example  4 : Percutaneous  Produce

A transdermal preparation is prepared with the following composition by using the propolis and the raw extract of the present invention containing the flavonoids.

[Furtherance]

Propolis and seed extract ............... 0.4 g

Sodium polyacrylate ...................... 1.3 g

Glycerin ................................. 3.6 g

Aluminum hydroxide ......................... 0.004 g

Methyl paraben 0.2 g

Acrylic adhesive solution 14 ml

Manufacturing example  5: Manufacture of functional food

Functional foods (beverages) are prepared with the following composition by using the propolis and the raw extract of the present invention containing the flavonoids.

[Furtherance]

Propolis and sprouts extract .................. 10 g

Citric acid ....................................... 1 g

Oligosaccharides ................................... 20 g

Honey ....................................... 10 g

Plum concentrate ................................. 2 g

Taurine ...................................... 1 g

Purified water was added to give 956 ml

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (9)

Wherein the extract comprises Propolis and Sophora japonica as an active ingredient, and the extract contains 5.0 to 20.0% by weight of flavonoid as an indicator substance.
The anti-inflammatory analgesic composition of claim 1, wherein the flavonoid is present in an amount of 5.0-15.0 wt%.
The anti-inflammatory analgesic composition according to claim 1, which exhibits anti-inflammatory analgesic effects against arthritis, otitis media, edema, pain or degenerative diseases caused by inflammation and inflammatory reaction.
The anti-inflammatory analgesic composition of claim 1, wherein the propolis and the granulation are in a weight ratio of 60:40 to 80:20.
Wherein the extract comprises Propolis and Sophora japonica as an active ingredient, and the extract contains 5.0 to 20.0% by weight of flavonoid as an indicator substance.
a) extracting with 2 to 4 times water, alcohol or alcohol aqueous solution, cooling and filtering to obtain a filtrate with a weight ratio of propolis and bean to 60:40 to 80:20;
b) adding sodium hydrogencarbonate to the filtrate to aggregate the waxy components of propolis at -20 ° C to -30 ° C and then filtering to obtain a filtrate from which wax is removed;
c) aging the filtrate for 1 to 7 days at room temperature to obtain a stock solution; And
d) centrifuging the stock solution at 3,000 ~ 5,000 rpm, and concentrating the filtrate under reduced pressure, freeze-drying, pulverizing and sterilizing the filtrate to obtain a propolis and a sprout extract
Wherein the flavonoid-containing propolis is selected from the group consisting of:
[7] The method according to claim 6, wherein the alcohol aqueous solution in step a) is an aqueous 60 to 80% ethanol solution.
[Claim 7] The method according to claim 6, wherein the propolis and the extract are in the form of powder.
The process according to any one of claims 6 to 8, wherein the flavonoid is from 5.0 to 20.0% by weight.

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