KR102007758B1 - Composition for alalgesic comprising high functional Capsicum frutescens IT 221919 extract - Google Patents

Abstract

The present invention relates to a painkiller composition containing a high functional pepper extract as an active ingredient and a food composition for pain improvement comprising the same. Highly functional red pepper (IT 221919) extract according to the present invention, compared to other red pepper extract omega-hydroxycapsaicin (omega-hydroxycapsaicin), homocapsaicin (homocapsaicin), capsaicin (capsaicin), dihydrocapsaicin (dihydrocapsaicin) and homodi Capsaicin analogues such as hydrocapsaicin (homodihydrocapsaicin) contains a large amount of TRPA1 (Transient Receptor Potential A1) inhibitory ability is very excellent and excellent analgesic effect.
Therefore, the present invention has a meaning in terms of excellent analgesic composition derived from natural products and can be used for the manufacture of medicines or health functional food and further contribute to national health.

Description

Composition for alalgesic comprising high functional Capsicum frutescens IT ２２１９１９ extract

The present invention relates to a painkiller composition and a pain improving food composition containing a high-performance pepper extract as an active ingredient.

Red pepper contains a lot of various vitamins and contains red pigment and spicy taste, making it a representative seasoning vegetable. Dried red pepper contains about 11%, 11%, 0.37% and 0.15% of protein lipid carbohydrate minerals and vitamins. In addition, pepper is known to contain a large amount of functional components of the flavonoid series (non-patent document 1) and functional components of the alkaloid series. These functional effects include preventing and eliminating obesity [Non-Patent Documents 2 and 3], strengthening cardiopulmonary function (Non-Patent Document 4) during exercise, and enhancing energy by increasing sex hormone metabolism function [Non-Patent Documents 5 and 6]. It is known to prevent dementia. Recent studies have shown that the preventive effect of dementia is to show the functional activity of anti-dementia by preventing the harmful effects of free radicals based on various antioxidants such as capsaicinoid, carotenoid, and flavonoids of red pepper. Patent Document 7]. Red pepper was also used as a herbal remedy. Chinese herbal medicine is named Buncho, Arabesque, Gocho, etc. In Chinese medicine, red pepper is a food that heats the heart and blood vessels. It is used as a topical stimulant for neuralgia and muscle pain. It is said to have been used for light frostbite, abdominal pain and stiff shoulders.

Currently, the Rural Development Administration is collecting and preserving core groups that can contain more than 99% of genetic diversity in order to improve pepper characteristics and develop new varieties. Based on the various resources of these peppers, research on increasing the agricultural production of peppers, as well as research on the functionalities of peppers that can improve human health or prevent from various diseases through daily intake, is needed.

Materska M, Piacente S, Stochmal A, Pizza C, Oleszek W, Perucka I. Isolation and structure elucidation of flavonoid and phenolic acid glycosides from pericarp of hot pepper fruit Capsicum annuum L., Phytochemistry 2003; 63 (8), 893-898) Lee G. R., Shin M. K., Yoon D. J., Kim A. R., Yu R., Park N. H. & Han I. S. Topical Application of Capsaicin Reduces Visceral Adipose Fat by Affecting Adipokine Levels in High-Fat Diet-Induced Obese Mice. Obesity 2013; 21 (1), 115-122 Lee M. S., Kim C. T., Kim I. H., & Kim Y. Effects of capsaicin on lipid catabolism in 3T3-L1 adipocytes. Phytother Res 2011; 25 (6), 935-939. Kang J. H., Goto T., Han I. S., Kawada T., Kim Y. M., & Yu R. Dietary capsaicin reduces obesity-induced insulin resistance and hepatic steatosis in obese mice fed a high-fat diet. Obesity 2010; 18 (4), page 780-787 Hirata N., Tokunaga M., Naruto S., Iinuma M., & Matsuda H. Testosterone 5alpha-reductase inhibitory active constituents of Piper nigrum leaf. Biol Pharm Bull 2007; 30 (12), 2402-2405 Vijayakumar R. S., & Nalini N. Piperine, an active principle from Piper nigrum, modulates hormonal and apo lipoprotein profiles in hyperlipidemic rats. J Basic Clin Physiol Pharmacol 2006; 17 (2), 71-86 Al-Baghdadi O. B., Prater N. I., Van der Schyf C. J., & Geldenhuys W. J. Inhibition of monoamine oxidase by derivatives of piperine, an alkaloid from the pepper plant Piper nigrum, for possible use in Parkinson's disease. Bioorganic & Medicinal Chemistry Letters 2012; 2 (23), 7183-7188

Accordingly, the present inventors have searched for resources exhibiting excellent functionality for pain relief among various pepper resources in Korea, and as a result, the present invention was completed by confirming the excellent effect of TRPA1 inhibitory ability on the IT 221919 pepper variety.

An object of the present invention is to provide an analgesic composition comprising red pepper IT 221919 extract exhibiting excellent functionality for pain relief as an active ingredient.

Another object of the present invention to provide a food composition for pain improvement comprising the pepper extract as an active ingredient.

Another object of the present invention to provide a health functional food for pain improvement comprising the pepper extract as an active ingredient.

As a means for solving the above problems, the present invention provides an analgesic composition comprising a red pepper extract of the National Institute of Agricultural Science, Agricultural Genetic Resource Center IT No. IT 221919 as an active ingredient.

As another means for solving the above problems, the present invention provides a food composition for pain improvement comprising the pepper extract as an active ingredient.

As another means for solving the above problems, the present invention provides a health functional food for pain improvement comprising the pepper extract as an active ingredient.

Highly functional red pepper (IT 221919) extract according to the present invention, compared to other red pepper extract omega-hydroxycapsaicin (omega-hydroxycapsaicin), homocapsaicin (homocapsaicin), capsaicin (capsaicin), dihydrocapsaicin (dihydrocapsaicin) and homodi Capsaicin analogues such as hydrocapsaicin (homodihydrocapsaicin) contains a large amount of TRPA1 (Transient Receptor Potential A1) inhibitory ability is very excellent and excellent analgesic effect. Therefore, the present invention is significant in terms of excellent analgesic composition derived from natural products and can be used for the manufacture of medicines or health functional food and further contribute to national health.

1 illustrates a pepper sample extraction and pretreatment process according to an embodiment of the present invention.
Figure 2 shows the results of high-speed mass screening of 75 peppers to discover TRPA1 inhibitory material.
Figure 3 is a result of analyzing five useful components contained in the pepper extract using LC-MS, it is comparing the relative content of omega-hydroxycapsaicin and homocapsaicin.
Figure 4 is a result of analyzing five useful components contained in the pepper extract using LC-MS, comparing the relative content of capsaicin, dihydrocapsaicin and homodihydrocapsaicin.

The present invention relates to a painkiller composition containing a high functional pepper extract as an active ingredient and a food composition for pain improvement comprising the same.

This will be described in more detail below.

The present inventors confirmed the analgesic activity of peppers of 75 different strains, and selected high functional peppers having excellent analgesic effect by analyzing the content of useful components.

"High-functional pepper" described in the present invention is Capsicum frutescens of the National Institute of Agricultural Science, IT Number IT 221919 line, means a pepper containing a large amount of useful components contained in the general pepper. The useful component includes omega-hydroxycapsaicin, homocapsaicin, homocapsaicin, capsaicin, dihydrocapsaicin, homodihydrocapsaicin and the like. In particular, the omega-hydroxycapsaicin (omega-hydroxycapsaicin) is included in 60 to 70 times higher content than other pepper strains. The homocapsaicin is included in 20-30 times higher content than other pepper strains. The capsaicin (capsaicin) is included in 5 to 7 times higher than other pepper strains. The dihydrocapsaicin (dihydrocapsaicin) is included 4 to 6 times higher than other pepper strains. The homodihydrocapsaicin (homodihydrocapsaicin) is included in 7 to 10 times higher than other pepper strains.

The red pepper extract may be obtained by extracting red pepper with at least one solvent selected from the group consisting of water and alcohols having 1 to 4 carbon atoms. In one embodiment, the red pepper extract may be obtained by extracting red pepper with 10 to 95% (v / v) alcohol having 1 to 4 carbon atoms, preferably 30 to 70% (v / v) methanol.

In order to sufficiently extract the active ingredient in the pepper, the amount of the extraction solvent used may be about 5 to 50 times, preferably about 10 to 20 times, based on the weight of the pepper, and extracted by sonication.

The composition of the present invention, in addition to the pepper extract as an active ingredient, nutrients, vitamins, electrolytes, flavors, coloring agents, neutralizing agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, Preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The components may be added independently or in combination. The content of the additional component is preferably added in the range of 0.01 to 100 parts by weight based on 100 parts by weight of the pepper extract, but is not limited thereto.

The content of the pepper extract in the composition can be appropriately adjusted according to the symptoms of the disease, the degree of symptoms, the condition of the patient, and the like, for example, 0.0001 to 99.9% by weight, preferably 0.001 to 50% by weight based on the total composition weight Preferably, but is not limited thereto. The content ratio is a value based on the drying amount from which the solvent is removed.

The composition may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions, and oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like according to conventional methods It may be used in the form of a dosage form, an external preparation, a suppository, or a sterile injectable solution.

In formulating the composition, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid form may include at least one or more excipients and / or lubricants. Liquid preparations for oral administration include suspensions, 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. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, and the like.

The preferred dosage of the composition depends on the condition and weight of the patient, the severity of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. For a more preferable effect, the dosage of the composition of the present invention is preferably 1 mg / kg to 100 mg / kg per day based on the active ingredient, but is not limited thereto. Administration may be administered once a day or may be divided several times. The compositions of the present invention can be administered by various routes to an animal, preferably a mammal, including a human. All modes of administration can be envisaged, for example, by oral, intravenous, intramuscular, subcutaneous injection or the like. The pharmaceutical dosage form of the composition of the present invention may be used in the form of a pharmaceutically acceptable salt of the active ingredient, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.

In addition, the present invention includes a food composition for pain improvement comprising the pepper extract.

In the present invention, the food means a natural product or processed product containing one or more nutrients, and preferably means a state in which it can be directly eaten through a certain processing process. , Dietary supplements, beverages, food additives and beverage additives. Examples of the foods include various foods, beverages, gums, teas, vitamin complexes, and functional foods. In addition, the food in the present invention includes special nutritional products (e.g., prepared oils, infants, baby food, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g. ramen, noodles, etc.), health supplements, seasoned foods ( For example, soy sauce, miso, red pepper paste, mixed soy sauce), sauces, confectionery (e.g. snacks), dairy products (e.g. fermented milk, cheese, etc.), other processed foods, kimchi, pickles (various kimchi, pickles, etc.), beverages ( Examples include, but are not limited to, fruits, vegetable drinks, soy milk, fermented beverages, ice cream, etc., natural seasonings (eg, ramen soup, etc.), vitamin complexes, alcoholic beverages, alcoholic beverages, and other health supplements. The health functional food, beverages, food additives or beverage additives may be prepared by a conventional manufacturing method.

The food composition may include food acceptable food additives, and may further include appropriate carriers, excipients, diluents, and the like commonly used in the manufacture of functional foods.

In addition, in the food, the amount of the red pepper extract may be included as 0.00001% to 50% by weight of the total food weight, if the food is a beverage based on 0.001 g to 50 g, based on 100 ml of the total food, Preferably, it may be included in a ratio of 0.01 g to 10 g, but is not limited thereto.

In addition, the present invention includes a health functional food comprising the pepper extract as an active ingredient.

The health functional food of the present invention may be prepared and processed in a form including, but not limited to, tablets, capsules, powders, granules, liquids, pills, and the like, for the purpose of pain improvement. The term "health functional food" used in the present invention refers to a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to the Health Functional Food Act No. 6767, regarding the structure and function of the human body It means the ingestion for the purpose of obtaining a beneficial effect for health use such as nutrient control or physiological action.

The health functional food of the present invention may include a conventional food additive, and the suitability as the "food additive" is applicable in accordance with the General Regulations of the Food Additives Code and General Test Methods, etc. approved by the Food and Drug Administration, unless otherwise specified. Judge according to the standards and standards for the item.

Examples of the items listed in the "Food Additive Revolution" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamon acid, natural additives such as color pigments, licorice extracts, crystalline cellulose, high color pigments, guar gum, Mixed preparations, such as a sodium L- glutamate preparation, an addition of an alkali, a preservative, and a tar coloring agent, etc. are mentioned.

For example, a dietary supplement in tablet form may be prepared by granulating a mixture of red pepper extract with excipients, binders, disintegrants, and other additives in a conventional manner, followed by compression molding with a lubricant, or by directly It can be compression molded. In addition, the health functional food in the form of tablets may contain a mating agent and the like, if necessary, may be coated with a suitable coating agent.

Hard capsules among the health functional foods in the form of capsules may be prepared by filling a hard capsule with a mixture of additives such as excipients or granules or peeled granules in a conventional hard capsule. It can be prepared by filling a mixture with an additive of a capsule base such as gelatin. The soft capsule agent may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, as necessary.

Ring-shaped dietary supplements can be prepared by molding mixtures of red pepper extracts, excipients, binders, disintegrants, etc. in a suitable manner. You may be greeted.

The health functional food in the form of granules may be prepared by granulating a mixture of the red pepper extract and the excipient, the binder, the disintegrating agent and the like in a suitable manner, and may contain a flavoring agent, a copper, and the like as necessary.

Terminology definitions for the excipients, binders, disintegrants, glidants, copulation agents, flavoring agents, etc. of the present invention are those described in literature known in the art and include those having the same or similar functions. Sacrament, Korean College of Pharmacy, 5 revised edition, p33-48, 1989).

Hereinafter, the configuration and effects of the present invention through the embodiments will be described in more detail. These examples are only for illustrating the present invention, but the scope of the present invention is not limited by these examples.

Example  1: pepper extract manufacturer

1) Pepper Ingredients Preparation

The pepper material used in the present invention was used by receiving the research material harvested in 2015 in connection with the pepper task currently underway at the Next Generation Bio Green 21 Project. Detailed information on pepper materials can be found in the Agricultural Life Resources Service (http://genebank.rda.go.kr/) operated by the National Agricultural Genetic Resource Center.

Pepper Resources Information number Experiment code IT No. Search site Search site URL Scientific name One D1 IT 221919 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. frutescens 2 D2 IT 135673 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 3 D3 IT 135674 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 4 D4 IT 158351 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 5 D5 IT 158680 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 6 D6 IT 158714 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 7 D7 IT 163532 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 8 D8 IT 223677 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 9 D9 IT 223737 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 10 D10 IT 236399 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 11 D11 IT 247232 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 12 D12 IT 158662 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 13 D13 IT 229667 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 14 D14 IT 237555 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 15 D15 IT 225019 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 16 D16 IT 218757 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 17 D17 IT 136602 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do Capsicum sp . 18 D18 IT 136633 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do Capsicum sp . 19 D19 IT 215521 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do Capsicum sp . 20 D20 IT 32379 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 21 D21 IT 236730 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 22 D22 IT 237617 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 23 D23 IT 158610 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 24 D24 IT 236341 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 25 D25 IT 240859 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 26 D26 IT 261138 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 27 D27 IT 229107 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 28 D28 IT 231151 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 29 D29 IT 225003 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 30 D30 IT 231168 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 31 D31 IT 258933 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do Capsicum sp . 32 D32 IT 199414 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 33 D33 IT 225013 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 34 D34 IT 235605 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 35 D35 IT 261417 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 36 D36 IT 223790 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 37 D37 IT 229133 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 38 D38 IT 229136 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 39 D39 IT 247244 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 40 D40 IT 158411 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 41 D41 IT 229185 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 42 D42 IT 231180 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 43 D43 IT 250191 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 44 D44 IT 250226 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 45 D45 IT 236754 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 46 D46 IT 237079 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 47 D47 IT 229152 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. baccatum 48 D48 IT 229148 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. baccatum 49 D49 IT 236738 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. chinense 50 D50 IT 229207 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. chinense 51 D51 IT 229211 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. chinense 52 D52 IT 284006 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. chinense 53 D53 IT 231135 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. frutescens 54 D54 IT 231141 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. frutescens 55 D55 IT 158583 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 56 D56 IT 158713 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. chacoense 57 D57 IT 294726 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 58 D58 IT 270425 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 59 D59 IT 158317 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. frutescens 60 D60 IT 158460 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. annuum 61 D61 IT 216362 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do Capsicum sp . 62 D62 IT 223623 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do Capsicum sp . 63 D63 IT 158570 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. frutescens 64 D64 IT 261550 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. baccatum 65 D65 IT 264149 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. baccatum 66 D66 IT 261486 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. baccatum 67 D67 IT 261540 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. baccatum 68 D68 IT 261510 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. baccatum 69 D69 IT 261524 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. baccatum 70 D70 IT 261512 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. baccatum 71 D71 IT 283938 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. baccatum 72 D72 IT 284013 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. baccatum 73 D73 IT 261158 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. baccatum 74 D74 IT 236721 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. chinense 75 D75 IT 261450 Agricultural Life Resource Service http://genebank.rda.go.kr/simpleSearch.do C. chinense

75 system pepper material of Table 1 was prepared.

2) pepper extract manufacture

The core fruits of the red peppers were pulverized and ground into grinders. The extracts were processed for functional evaluation and component analysis without standardization of particle size. 0.5 g of red pepper powder samples were added to the vial, 70% MeOH was added at a concentration of 1 g / 10 mL, and then sonicated twice for 30 minutes to extract metabolites. After centrifugation at 1,000 rpm for 5 minutes, the supernatant was filtered using a 0.45 um syringe filter, transferred to a new vial weighed, and concentrated with a nitrogen concentrator to completely remove water. The concentrated sample was re-dissolved in DMSO at a concentration of 40 mg / mL, and the activity analysis and the component analysis were performed after filtration using a 0.2 um syringe filter (FIG. 1).

Samples for component analysis were diluted to 5 mg / mL concentration with 100% MeOH.

Example  2: using fast bulk screening TRPA1  Exploration control

Transient Receptor Potential A1 (TRP A1), a cation channel expressed in various tissues such as nerve cells, skin, respiratory organs, and kidneys, is a receptor that is activated by external stimulation and chemical stimulation sources and stimulates cells by introducing cations into cells. Was performed to high-speed mass-screening using the Fluo-4 TRPA1 Direct ^TM binding to stable cell lines expressing the Ca ^{2 +} to identify materials which inhibit the activity of TRPA1 by irritants.

Place 100 µl of TRPA1 H10 cells (stable cell line for mass screening of TRP A1 inhibitors produced by KISA) on a 96 well black clear plate and place 100 µl of 4 X 10 ⁴ cells / well for 1 minute at room temperature at 1,000 rpm. After centrifugation, the cells were incubated for 48 hours in a 37 ° C., 5% CO ₂ incubator (medium: DMEM, 10% FBS, 1% Penicillin-Streptomycin, 500 μg / ml G418).

^{Fluo-4 Direct TM Calcium assay kit} (Thermo Fisher Scientific F10472) Probenecid (Component B) Fluo-4 Direct TM Calcium assay buffer (Component C) contained in the 77mg 1vial into 1㎖, it was Vortex mix well (final concentration of 250 mM probenecid).

^{Fluo-4 Direct TM Calcium assay Reagent} (Component A) 10 ㎖ of ^{Fluo-4 Direct TM Calcium assay buffer} (Component C) and the 200 ㎕ 250 mM Probenecid (Component B ) a rear semi-put, and then mix well (2 X in Fluo-4 Direct ^TM calcium reagent) and 100 μl of 2 × Fluo-4 Direct ^™ calcium reagent were added to the cells and allowed to react for 1 hour in a 37 ° C. incubator.

In order to confirm the effect of agonist, Calcium assay was performed using Polygodial, and Calcium assay protocol [fluorescence at EX. 495 nm / Em. 516, 140s read, read 67, 25s agonist 25μl injection (Flexstation)].

All reagents and protocols used above were used according to the reagents and methods included in the Fluo-4 Direct ^™ Calcium assay kit, catalog number F10472 from Thermo Fisher Scientific.

As a result of measuring the inhibition of TRPA1 using a total of 75 pepper extracts, the inhibition rate was in the range of 0 to 54.4%, and the highest inhibitory activity was confirmed in the IT 221919 pepper strain. The average TRPA1 inhibition rate and standard deviation for the 75 pepper extracts analyzed were 11.3 ± 10.9%, and the most active IT 221919 strain showed enhanced analgesic activity of about 43% more than the average.

Example 3 LC-MS Analysis of Five Useful Components of Red Pepper Extract

Five useful ingredients contained in the red pepper extract (omega-hydroxycapsaicin, homocapsaicin, capsaicin, dihydrocapsaicin and homodihydrocapsaicin) The analysis was performed using LC-MS as follows.

Analytical Instruments: Nexera X2 UHPLC (Shimadzu)

Q-TOF-MS (TripleTOF 5600 ⁺ system, AB SCIEX)

Analytical column: Shim-pack GIS-ODS column (3 μm, 3.0 × 100 mm)

-Detector: Diode-array detector (DAD) & MS detector

Solvent Conditions: Mobile Phase A-0.1% formic acid in water

           Mobile phase B-0.1% formic acid in 100% acetonitrile

UHPLC analysis conditions:

Q-TOF / MS analysis conditions:

The five useful components contained in the pepper extract were analyzed using LC-MS, and the results are shown in FIGS. 3 and 4.

The distribution of the contents of capsaicin, dihydrocapsaicin and homodihydrocapsaicin in the strains showed a certain pattern. However, in the IT 221919 line, capsaicin and dihydrocapsaicin were not significantly different from some hot peppers, while the omega-hydroxycapsaicin and homocapsaicin contents were significantly higher than those of other lines. The pepper strain IT 221919 showed a peak area value of about 68.4 times higher than that of the rest of the 74 strains in the content of omega-hydroxycapsaicin and 27.4 times higher in the homocapsaicin content. On the other hand, capsaicin, dihydrocapsaicin and homodihydrocapsaicin were found to be 5.7 times, 5.0 times and 8.6 times higher than the average of 74 lines except IT 221919, respectively.

Therefore, the extract of pepper IT 221919 strain used in the present invention was higher in content of omega-hydroxycapsaicin, homocapsaicin, homodihydrocapsaicin, capsaicin and dihydrocapsaicin than other strains.

Claims (11)

It contains pepper extract of IT No. IT 221919, National Institute of Agricultural Science,
The pepper extract is obtained by extracting pepper with at least one solvent selected from the group consisting of alcohols having 1 to 4 carbon atoms of 10 to 95% (v / v), analgesic composition.
delete The method of claim 1,
The pepper extract is obtained by extracting pepper with 10 to 95% (v / v) methanol, analgesic composition.
The method of claim 1,
The pepper extract contains one or more selected from the group consisting of omega-hydroxycapsaicin, homocapsaicin, homocapsaicin, capsaicin, dihydrocapsaicin, and homodihydrocapsaicin That is, painkiller composition.
The method of claim 4, wherein
The omega-hydroxycapsaicin (omega-hydroxycapsaicin) is included in 60 to 70 times higher content than other pepper strains, analgesic composition.
The method of claim 4, wherein
The homocapsaicin (homocapsaicin) is included in 20 to 30 times higher content than other pepper strains, analgesic composition.
The method of claim 4, wherein
The capsaicin (capsaicin) is contained in a 5 to 7 times higher content than other pepper strains, analgesic composition.
The method of claim 4, wherein
The dihydrocapsaicin (dihydrocapsaicin) is included in 4 to 6 times higher content than other pepper strains, analgesic composition.
The method of claim 4, wherein
The homodihydrocapsaicin (homodihydrocapsaicin) is contained in a 7 to 10 times higher content than other pepper strains, analgesic composition.
It contains pepper extract of IT No. IT 221919, National Institute of Agricultural Science,
The pepper extract is obtained by extracting pepper with at least one solvent selected from the group consisting of alcohols having 1 to 4 carbon atoms of 10 to 95% (v / v), food composition for pain improvement.
It contains pepper extract of IT No. IT 221919, National Institute of Agricultural Science,
The pepper extract is obtained by extracting the pepper with at least one solvent selected from the group consisting of alcohols having 1 to 4 carbon atoms of 10 to 95% (v / v), health functional food for pain improvement.

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