KR101851010B1 - Compositions for the antiinflammatory and the antimicrobial activities - Google Patents

Abstract

R = H or OHThe present invention provides a composition having an anti-inflammatory and antimicrobial effect containing the following general formula (I) as an active ingredient.
The compound of formula (I)

R = H or OH

Description

Compositions for antiinflammatory and antimicrobial activities [0002]

The present invention relates to derivatives of isoflavone compounds, and more particularly to compositions having anti-inflammatory and antimicrobial properties.

The human body suffers from many kinds of inflammatory diseases throughout its life. In particular, chronic irritable inflammation such as atopic dermatitis, allergic dermatitis, rheumatoid arthritis, bronchial asthma, and acne have not been established so far, so it is imperative to develop new drugs that can control and treat these diseases .

Inflammation is one of the defenses of biological tissue against external stimuli from cells and tissues. It causes cell reactions with nerves, blood vessels, tissues, blood and the like. As a result, if it is excessive, it may cause itching, fever, And the like. The inflammatory response begins by recognizing the pattern of cell surface that is specific to the pathogen as part of the innate immune response. Macrophages recognize cells with such specifically present cell surfaces as non-magnetic and attach to pathogens. The cause of inflammation is related to various biochemical phenomena. In particular, biosynthesis of nitric oxide synthase (NOS), which is an enzyme that produces nitric oxide (NO), and various prostaglandins (PGs) The mediating enzyme cyclooxygenase (COX) is an important mediator of the inflammatory response. Since NO and PGs are closely related to the inflammatory reaction, a substance capable of regulating the expression of the enzyme involved in the production and production of NO and PGs is attracting attention as a preventive and therapeutic agent for inflammatory diseases. Until now, the substances that have been used for antiinflammatory purposes are nasal steroids such as ibuprofen, indomethacin, etc. Steroids include prednisolone, dexamethasone, Antagonists, bradykinin antagonists, COX inhibitors and the like have been proposed. However, they have problems in terms of safety for skin and stability in formulation, and their use is limited.

Skin diseases called acne are skin diseases that occur in the hair follicles that show various skin lesions such as obstructive or open skin, papules, pustules, cysts, and nodules.

The causes of acne include increased secretion of sebum, abnormal keratinization in follicles, and community formation of acne bacteria. These factors work together to cause acne lesions. That is, when the function of the sebaceous gland is enhanced by the secretion of the male hormone and the like, and the excess sebum generated thereby causes the follicular wall overgrowth and stagnates in the hair follicle, the follicle is formed, which is an early stage of the onset of acne. As the secretion of sebum increases and the hair follicle is excessively follicularized, sebaceous stasis in the hair follicles blocks the air circulation by blocking the hair follicle, which causes an environment in which the acne bacterium that resides in the hair follicle can grow well, do. In this case, acne bacterium secretes lipolytic enzymes and chemotactic factors to make free fatty acids and white blood cells around the hair follicle, and they stimulate and destroy the hair follicles, the contents of the hair follicles into the dermis into the early non-inflammatory acne inflammatory acne And it is exacerbated by purulent acne.

In general, the treatment of acne is accomplished by suppressing the cause of its occurrence. Examples of topical application agents used for treating acne by removing the above-mentioned acne bacteria include benzoyl peroxide, azelaic acid, retinoic acid, salicylic acid, Sulfur, topically applied antibiotics, and the like. However, these preparations do not exhibit sufficient antibacterial effect against acne bacterium only with a small amount, and there is a problem that side effects such as skin eruption or skin dryness may occur during excessive application.

Disclosure of Invention Technical Problem [8] The present invention has been made to solve the problems of the prior art as described above, and its object is to provide a composition which has no side effects on human body and is excellent in anti-inflammatory and antibacterial effects.

The technical problem of the present invention as described above is achieved by the following means.

(1) an active ingredient selected from the group consisting of 6,8-diprenylorobol, 6,8-diprenylgenistein and mixtures thereof, ≪ RTI ID = 0.0 > anti-inflammatory < / RTI >

(2) The method according to claim 1, wherein 6,8-diprenylorobol, 6,8-diprenylgenistein and mixtures thereof are added to the entire composition ≪ / RTI > by weight, based on the total weight of the composition.

(3) The method according to claim 1, wherein the 6,8-diprenylorobol, 6,8-diprenylgenistein and mixtures thereof are obtained from the tree ≪ / RTI >

(4) The pharmaceutical composition according to (1), wherein the anti-inflammatory activity is selected from the group consisting of various dermatitis, skin irritation, atopic eczema, acne, arthritis, conjunctivitis, rhinitis, hypertension, diabetes, allergy, cancer, arteriosclerosis, Diabetes, dementia, depression, schizophrenia, tuberculosis, sleep disturbance, sepsis, burns, gingivitis, gingivitis, inflammatory bowel disease, gout, myocardial infarction, congestive heart failure, hypertension, angina, gastric ulcer, cerebral infarction, Down syndrome, multiple sclerosis Or < / RTI > pancreatitis.

(5) The composition according to claim 1, wherein the antimicrobial activity is characterized by the prevention and treatment of acne, dandruff, deodorant, hair loss, athlete's foot and cavities.

(6) The composition according to claim 1, wherein the composition is a cosmetic composition, a pharmaceutical composition or a health functional food.

7. The cosmetic composition according to claim 6, wherein the cosmetic composition is at least one selected from the group consisting of a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, A nutritional essence, a pack, a soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion and a body cleanser.

(8) The composition according to claim 6, wherein the pharmaceutical composition is a cream, a gel, a patch, a spray, an ointment, a warning agent, a lotion, a liniment, a pasta or a cataplasma.

(9) The composition according to claim 6, wherein the health functional food is a health functional food, which is powder, granule, tablet, capsule, syrup, or beverage.

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According to the present invention, it is possible to provide a composition having no side effects on the human body and having excellent anti-inflammatory and antibacterial effects.

The present invention provides an isoflavone derivative compound having a chemical structure of the following general formula (I) having excellent anti-inflammatory and antimicrobial effect or a pharmacologically acceptable salt thereof:

The compound of formula (I)

Wherein R is hydrogen or a hydroxy group.

Examples of the compound of the general formula (1) include 6,8-diprenylorobol and 6,8-diprenylgenistein.

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

The compound of the present invention may be isolated from Cudrania tricuspidata extract or synthesized by biological or chemical methods. As a method of extracting the above compounds, known methods can be used. For example, the extract is extracted from an finely pulverized plant material using an organic solvent, dried, suspended in water, extracted with liquid-liquid And a method of obtaining the compound of the present invention purely by column chromatography and high-performance liquid chromatography.

The preferred dosage of the compound of formula (I) of the present invention or its pharmacologically acceptable salt varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, . However, for the desired effect, the compound of the present invention is preferably administered at 0.0001 to 100 mg / kg per day, preferably 0.001 to 10 mg / kg 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 compounding amount of the compounds according to the present invention in the above composition is preferably 0.0001 to 30% by weight, more preferably 0.01 to 20% by weight based on the total weight of the composition. If the amount is less than 0.01% by weight, If it is more than 20% by weight, it is difficult to expect an increase in the effect even if it is blended.

Since the isoflavone derivatives of the present invention have been conventionally used as food additives, they are safe without side effects even if they are used in the human body.

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

The composition comprising the compound of the general formula (I) of the present invention or a pharmacologically acceptable salt thereof as an active ingredient can be used for various inflammatory diseases such as atopic dermatitis, arthritis and allergic dermatitis and microbial diseases such as acne, dandruff, The present invention also relates to a pharmaceutical composition for preventing and treating cancer.

The components contained in the composition of the present invention include components commonly used in the composition other than the above isoflavone derivative compounds as an active ingredient and can be used as a conventional auxiliary agent such as antioxidants, stabilizers, solubilizers, vitamins, And a carrier.

The composition of the present invention may be prepared in any formulations conventionally produced in the art and may be in the form of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, A powder foundation, an emulsion foundation, a wax foundation and a spray, but the present invention is not limited thereto. More particularly, when the composition of the present invention is used as a cosmetic composition, it is formulated into a form of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder .

When the formulation of the present invention is a paste, a cream or a gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, chlorofluorohydrocarbons, propane / Or propellants such as dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butylene glycol, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

When the formulation of the present invention is a suspension, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide ether sulfate, alkyl Aliphatic alcohols, fatty acid glycerides, fatty acid diethanol amides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters.

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

[Example 1] Preparation of 6, 8-diprenylorobol

100 g of the ground powder was placed in 500 ml of ethanol, boiled for 3 hours in a reflux condenser equipped with a condenser, filtered through 300 mesh filter cloth, and the residue was extracted once more by the same method. The extracts were combined and filtered through Hattman filter paper at room temperature to remove insoluble matter. The extract was dried under reduced pressure in a distillation apparatus equipped with a cooling condenser, suspended in 300 ml of purified water, 300 ml of hexane was added thereto and shaken to obtain hexane lysate. The hexane lysate was dried under reduced pressure and purified by silica gel column chromatography (developing solvent: ethyl acetate / n-hexane = 1 / 10-1 / 1) to obtain 39 mg of yellow needle crystal compound. The obtained compound was analyzed by electrospray ionization mass spectrometry (hereinafter referred to as ESI-MS) and hydrogen nuclear magnetic resonance analysis (hereinafter referred to as ¹ H-NMR) to obtain 6,8-diprenylowol (6, 8-diprenylorobol).

ESI-MS: m / z 423 [M + H] +, 421 [MH] -

_{^{1 H-NMR (CDCl 3,}} 300MHz): δ 7.87 (s), 6.95 (d, J = 1.1), 6.80 (d, J = 8.1Hz), 6.73 (dd, J = 1.1Hz, 8.1Hz), 3.43 (m), 5.23 (m) , 1.83 (s), 1.75 (s), 3.45 (m), 5.21 (m), 1.73 (d, J = 0.9Hz), 1.82 (d, J = 0.9Hz)

[Example 2] Preparation of 6,8-diprenylgenistein

23 mg of yellow needle crystals were obtained in the same manner as in Example 1 except that the developing solvent of the silica gel column chromatography was replaced with ethyl acetate / n-hexane = 1 / 20-1 / 2). The obtained compound was analyzed by electrospray ionization mass spectrometry (hereinafter referred to as ESI-MS) and hydrogen nuclear magnetic resonance analysis (hereinafter referred to as ¹ H-NMR) to obtain 6,8-diprenyl genistein (6, 8-diprenylgenistein).

ESI-MS: m / z 407 [M + H] +, 405 [MH] -

_{^{1 H-NMR (CDCl 3,}} 300MHz): δ 7.82 (s), 7.26 (d, J = 8.5), 6.77 (d, J = 8.5Hz), 6.77 (d, J = 8.5Hz), 3.40 (m) , 5.18 (m), 1.76 ( d, J = 1.1Hz), 1.69 (d, J = 1.1Hz), 5.14 (m), 1.66 (d, J = 0.7Hz), 1.75 (d, J = 0.7Hz)

[Experimental Example 1] NO production inhibitory effect

In order to confirm the anti-inflammatory effect of 6,8-diprenyl oulicol and 6,8-diprenyl genistein prepared in Examples 1 and 2, a nitric oxide (NO) production inhibitory effect test Respectively. (A. J. Ding, C. F. Nathan, and D. J. Stuehr, J. Immunol., 144, 2407-2413, 1988)

RAW 264.7 cells, a macrophage of mice, were cultured in 10% FBS DMEM medium and cultured on a 24-well plate at 4 × 10 ⁵ cells / well for 24 hours in a 5% CO ₂ thermostat. After the medium was removed, the cells were treated with FBS free DMEM containing the compounds of Examples 1 and 2 and LNMMA, respectively. After 30 minutes, NO was induced with LPS (lipopolysaccharide) and cultured for 24 hours. NO assay kit was used to calculate the NO level of the sample compared to the LPS treatment, and the inhibition rate of NO production was evaluated. The results are shown in Table 1 below.

NO production inhibition rate sample Concentration (ppm) NO production inhibition rate (%) LNMMA 10 46.2 6,8-diprenylorobole 10 60.1 20 102.3 6,8-diprenylgenistein 10 60.5 20 104.2

As shown in Table 1, the compounds of the present invention showed very strong inhibition of NO production, indicating excellent anti-inflammatory activity. Compounds of the present invention showed much better results than LNMMA, a typical NO production inhibitor.

[Experimental Example 2] Measurement of anti-acne effect

The antibacterial effects of the 6,8-diprenyl oulovol and 6,8-diprenyl genistein prepared in Examples 1 and 2 were tested. Propionibacterium acnes ATCC 6919, an acne bacterium, was used as an isolate.

The minimum inhibitory concentration (MIC) assay was used to measure the antimicrobial activity. Each sample was diluted at a fixed concentration interval in the liquid medium, and then each test strain was inoculated and cultured. Then, the minimum concentration to inhibit the growth of the microorganism was determined by observing the growth of the microorganism by the sample concentration. The growth of the bacteria was determined by adding 1% TTC (2,3,5-Triphenyl Tetrazolium Chloride) solution and the growth was positive (+) when the color changed from colorless to red, and negative (-) when the color was not changed. TTC is used to easily measure the viability of a microorganism since it is a dye that turns red when it is reduced in its original oxidized state due to the change of redox potential which occurs when the bacteria growth is active. Propionibacterium acnes, a causative agent of acne, is an anaerobic strain. Therefore, after the inoculation, it was incubated in an anaerobic chamber at 37 ℃ for 2 days. The results obtained are shown in Table 2 below.

Antimicrobial activity against acne bacteria sample MIC (ppm) Triclosan 5 to 10 6,8-diprenylorobole 5 to 10 6,8-diprenylgenistein 5 to 10

Referring to Table 2, in the results of MIC measurement for acne bacteria, the compounds of the present invention exhibited the same or higher antibacterial activity as those of triclosan, which is a typical acne inhibitor.

In Experimental Example 1 of the present invention, the effect of 6,8- diprenyl ouloprobe and 6,8- diprenyl genistein on the anti-acne bacteria was tested. Malassezia furfur was applied to dandruff, Corynebacterium xerosis was applied to the dorsum , The Minimum Inhibitory Concentration (MIC) assay, which is the same method as in Experimental Example 1, was conducted using Staphylococcus epidermidis against hair loss, Trichophyton mentagrophytes against athlete's foot, and Streptococcus mutans against cavities, respectively Can be used to measure the antibacterial activity. In other words, each sample can be diluted at a fixed concentration interval in a liquid medium, incubated with each of the test strains, and then examined for the growth rate of the cells by the sample concentration. At this time, if the color changes from colorless to red, 1% TTC (2,3,5-Triphenyl Tetrazolium Chloride) solution is added, and the growth is positive (+). The antimicrobial activity of the composition according to the present invention can be easily confirmed by the above method.

[Production Examples 1 and 2 and Comparative Example 1]

Nutrition lotions of Examples 1 and 2 and Comparative Example 1 were prepared by the ingredients and contents shown in Table 3 below.

Manufacture of nutrition lotion Composition (% by weight) Manufacturing example Comparative Example 1 One 2 Lactic acid 3  3   3 6,8-Diprenyl Orobor 0.1 - - 6,8-Diprenyl Genistein   - 0.1   - Polyoxyethylene hardened castor oil 1.0 1.0 1.0 Methyl paraoxybenzoate 0.2 0.2 0.2 glycerin 6.0 6.0 6.0 1,3-butylene glycol 5.0 5.0 5.0 Carbomer 0.2 0.2 0.2 Triethanolamine 0.3 0.3 0.3 Propylene glycol 5.0 5.0 5.0 ethanol 3.2 3.2 3.2 Carboxyvinyl polymer 0.1 0.1 0.1 incense Suitable amount Suitable amount Suitable amount Purified water to 100 to 100 to 100

 [Experimental Example 3] Measurement of skin irritation mitigation effect

In general, how the compounds according to the present invention alleviate skin irritation caused by lactic acid, which is known to stimulate the skin, is examined. The compositions of Examples 1 and 2 and Comparative Example 1 were applied to each of 15 female rubbers by rubbing the composition of each of Examples 1 and 2 and Comparative Example 1 by a spatula in an amount of 20 ul each time. At this time, the first two times were applied to the hill top chamber without the filter paper in the inside after the application of the composition, and the closure conditions were established by fixing with an adhesive. In the chamber reaction by the hilltop chamber, when the composition was applied three to eight times, the filter paper was placed immediately after application and fixed with an adhesive. Spreaded once every 23 hours, and inspected 1 hour after removal of the patches according to the CTFA guidelines. On the 4th day after the end of the patch, an additional test was performed to see the delayed type response and the result was reflected in the results. Data were analyzed by post-test (p <0.05) using ANOVA after confirming equal distribution of Levin test using the last test result. The results were determined according to the measurement criteria shown in Table 4 below and the results are shown in Table 5 below. The results shown in Table 5 are summarized by summing up the response indices of the subjects in response to the stimulus by the number of times.

Criteria for skin irritation mitigation effectiveness Rating sign condition 0 - No visible response One + Weak erythema 2 ++ Strong erythema 3 +++ Strong erythema and edema 4 ++++ Strong erythema, edema, blisters

The response index of the subject to the number of patches Number of Patches Production Example 1 Production Example 2 Comparative Example 1 One 0 0 One 2 One 0 6 3 0 0 3 4 0 0 4 5 0 One 3 6 0 0 5 7 0 One 3 8 One One 4 Four days after the end One One 9 Sum 3 4 38

As shown in Table 5, in the case of the composition of Comparative Example 1, the stimulation reaction to skin by lactic acid was very high, while the 6,8-diprenylol ball of the present invention and 6,8-diprenyl It was found that the compositions of Examples 1 and 2 containing genistein had a relatively low skin irritation by lactic acid.

[Production Examples 3 to 4 and Comparative Example 2]

Creams of the following Examples 3 and 4 and Comparative Example 2 were prepared by the ingredients and contents shown in Table 6 below.

Manufacture of cream Composition (% by weight) Example Comparative Example 2 3 4 6,8-Diprenyl Orobor 0.1 - - 6,8-Diprenyl Genistein   - 0.1   - Stearic acid 15 15 15 Cetanol One One One Potassium hydroxide 0.7 0.7 0.7 glycerin 5.0 5.0 5.0 Propylene glycol 3 3 3 antiseptic Suitable amount Suitable amount Suitable amount incense Suitable amount Suitable amount Suitable amount Purified water to 100 to 100 to 100

[Experimental Example 4] Test for improving atopic symptoms in the human body

In order to confirm the effect of improving the atopic symptoms by the composition of the present invention, the degree of improvement of atopic dermatitis was evaluated by applying the creams prepared in Examples 3 and 4 and Comparative Example 2. Twenty-four panels with symptoms of dermatitis exhibiting symptoms similar to each other were divided into three groups of eight each, and the compositions of Examples 3 and 4 and Comparative Example 2 were applied twice daily for eight weeks to the face. At the end of the test, subjects were asked to fill out a questionnaire and conducted a subjective efficacy evaluation. In the questionnaire, symptoms of atopic dermatitis were divided into canopy, dryness, keratinization, dandruff, erythema, swelling, skin cracking, dirt and eczema, And the effect of improving atopic symptoms was evaluated by averaging them. The results are shown in Table 7 below.

Atopic symptom improvement effect result Degree of improvement Example 3 Example 4 Comparative Example 2 Significant improvement 7 6 0 Improving One 2 2 No change 0 0 5 worse 0 0 One

As shown in Table 7, when the compositions of Examples 3 and 4 containing 6,8-diprenyl oulobate and 6,8-diprenyl genistein of the present invention were applied, symptoms of atopic dermatitis were improved .

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. It can be understood that

Claims (9)

Acne containing an effective ingredient selected from the group consisting of 6,8-diprenylorobol, 6,8-diprenylgenistein, and mixtures thereof, Antimicrobial composition for prevention and treatment. The composition according to claim 1, wherein 6,8-diprenylorobol, 6,8-diprenylgenistein and mixtures thereof are present in an amount ranging from 0.0001% &Lt; / RTI &gt; 30.0% by weight of the composition. The method according to claim 1, wherein the 6,8-diprenylorobol, 6,8-diprenylgenistein and mixtures thereof are derived from a tree . delete delete The composition according to claim 1, wherein the composition is a cosmetic composition, a pharmaceutical composition or a health functional food. The cosmetic composition according to claim 6, wherein the cosmetic composition is at least one selected from the group consisting of a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutrition cream, a moisturizing cream, a hand cream, Pack, a soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion and a body cleanser. The composition according to claim 6, wherein the pharmaceutical composition is a cream, a gel, a patch, a spray, an ointment, an alarm, a lotion, a liniment, a pasta or a cataplasma. The composition according to claim 6, wherein the health functional food is a health functional food which is powder, granule, tablet, capsule, syrup or beverage.