KR20090064865A - Skin inflammation, disease and skin tumor preventive agent containing 5-deoxykaempferol - Google Patents

Abstract

An agent for preventing and treating skin inflammation, skin disorder, and skin cancer is provided to inhibit the expression of cyclooxygenase-2(COX-2) which is increase by the irradiation of ultraviolet irradiation. An agent for preventing and treating skin inflammation, disorder and cancer comprises 0.1-50 weight% of deoxykaemferol as an active ingredient. The effective amount is 10 muM-1mM. The composition is used in medicine. The medicine is anticancer drug, and has anticancer effect. The composition is used in a form of food product, food additive, beverage or beverage additive.

Description

Skin inflammation, skin disease and skin cancer prevention and treatment including 5-deoxy camphorol {SKIN INFLAMMATION, DISEASE and SKIN TUMOR PREVENTIVE AGENT CONTAINING 5-DEOXYKAEMPFEROL}

The present invention relates to a composition for preventing and treating skin inflammation, skin disease and skin cancer comprising 5-deoxycamphorol (3,7,4'-trihydroxyflavone, resokaempferol) as an active ingredient among flavonoid compounds. For example, 5-deoxycampol, which has an effect of increasing cyclooxygenase-2 (COX-2) expression by UV rays and inhibits skin cancer, is effective in preventing and treating skin inflammation, skin damage and skin cancer. It relates to a prophylactic and therapeutic composition for use.

Recently, the immune system of our body is disturbed by the exposure of environmental pollution, the increase of stress, and the change of diet of the modern people, and thus various skin diseases are increasing rapidly.

Sun light is composed of visible, ultraviolet and infrared light. UV rays synthesize vitamin D in the body and play a role in sterilization, but may also cause dermatitis, skin cancer, sunburn, skin aging, drying, and fine lines. Ultraviolet (UV) is divided into three types, A, B, and C, depending on the wavelength, among which UVC is blocked in the ozone layer, and UVA and UVB affect the skin.

In particular, the medium wavelength UVB (290-320 nm) causes severe skin damage when exposed to acute or chronic exposure, causing sunburn, mainly causing inflammation of the skin and erythema or blisters. Overexposure is known to cause skin inflammation and cancer (Oro, AE et al., Science , 276: 817-821, 1997; Kim HJ et al., Apoptosis 9: 449-456, 2004).

When excessively inflammatory substances are produced, it may cause excessive immune response, causing various inflammatory diseases and skin damage. Cyclooxygenase-2 (hereinafter referred to as COX-2), an enzyme that is important in inflammation, is a representative enzyme that produces arachidonic acid (prostaglandins) as an inflammatory substance (Hla T and Neilson K. Proc Natl Acad Sci USA . 89 (16): 7384-8, 1992, Grewe M et al., J Invest Dermatol . 101 (4): 528-31, 1993; Joyce E. Rundhaug et al., Mol Carcinog . , 46: 692-698, 2007). Recent studies have also reported increased COX-2 activity in several types of carcinoma (Dubois RN, FASEB J , 12: 1063-73, 1998, Brecher AR., J Drugs Dermatol ., 1 (1): 44-7, 2002). Accordingly, there is a need for development of a substance capable of inhibiting COX-2 expression as an agent for preventing and treating skin inflammation, skin damage and skin cancer.

The present invention solves the problems of the prior art and is made by the necessity, it is an object of the present invention to provide a skin inflammation and skin disease prevention and treatment.

In order to achieve the above object, the present invention comprises 5-deoxycamphorol (3,7,4'-trihydroxyflavone, resokaempferol) represented by the following formula 1 as an active ingredient, preventing skin inflammation, skin disease and cancer prevention and Provided are pharmaceutical compositions of treatment.

(Formula 1)

The inventors of the present invention, while studying the prevention and treatment of skin inflammation to ensure human safety, it was confirmed that 5-deoxycampol can effectively suppress the COX-2 expression and skin cancer increased by ultraviolet light, based on this The invention was completed.

The present invention relates to a skin inflammation, skin disease and skin cancer prevention and treatment agent comprising 5-deoxycamphor as an active ingredient. 5-deoxycampolol is a commercially available compound.

The 5-deoxycamphorol content included in the skin inflammation, skin disease and cancer prevention and treatment of the present invention is preferably controlled according to the method of use of the prevention and treatment, the condition of the user, the type of disease and the severity of the disease. Good to do. An effective amount of 5-deoxycamphor in the composition of the present invention may be 0.01 to 50% by weight, but is not limited thereto. However, if the content is less than 0.01% by weight may be ineffective anti-inflammatory and skin damage prevention, if the content exceeds 50% by weight can be uneconomical low effect increase rate.

In addition, in one embodiment of the present invention, the effective amount is preferably 10μM to 1mM, but is not limited thereto .

Skin inflammation, skin disease and skin cancer prevention and treatment of the present invention, in addition to the active ingredient may further comprise a pharmaceutically acceptable carrier, diluent or excipient, wherein the content of the active ingredient is 0.001 to 99% by weight of the total composition It is good. Carriers, excipients or diluents which may be used include lactose, dextrose, sucrose, sorbitol, mannitol, xyitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, which may be used one or more. In addition, when the prophylactic and therapeutic agent is a medicament, it may further include fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers or preservatives.

In addition, the skin inflammation, skin disease and skin cancer prevention and treatment agent comprising as an active ingredient of the present invention can be used as a drug, food, food additives, beverages, or beverage additives. The cancer prevention composition may be a cancer prevention agent when used as a medicament, and when used as a food, a food additive, a beverage or a beverage additive, various foods, meats, drinks, chocolate, snacks, confectionery, pizza, ramen, other noodles, gums , Ice creams, alcoholic beverages, vitamin complexes, alcoholic beverages and other health supplements, but is not limited thereto. In this case, the cancer prevention composition may be included in 0.001 to 50% by weight in the final drug, food or beverage, but is not limited thereto.

Formulations of prophylactic and therapeutic agents for skin inflammation, skin disease and skin cancer may be in the preferred form depending on the method of use, and in particular methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. It is good to adopt and formulate. Examples of specific formulations include PLASTERS, GRANULES, LOTION, LPTIONS, LINIMENTS, LIMONADES, AROMATIC WATERS, POWDERS, Syrup ( SYRUPS), OPHTALMIC OINTMENTS, LIQUIDS AND SOLUTIONS, AEROSOLS, EXTRACTS, ELIXIRS, OINTMENTS, FLUIDEXTRACTS, Emulsion ), SUSPENSIONS, DECOCTIONS, INFUSIONS, OPHTHALMIC SOLUTIONS, TABLETS, Suppositories (SUPPOSITIORIES), INJECTIONS, SPIRITS, CATASLSMA ), Capsules (CAPSULES), creams (CREAMS), troches (TROCHES), tinks (TINCTURES), pasta (PASTES), pills (PILLS), soft or hard gelatin capsules.

The dosage of the skin inflammation, skin disease and skin cancer prevention and treatment agent according to the present invention may be determined in consideration of the method of administration, the age, sex and weight of the recipient, and the severity of the disease. For example, the skin inflammation, skin disease and skin cancer prevention and treatment of the present invention can be administered one or more times 0.1 to 100 mg / kg (body weight) per day based on the active ingredient. However, the above dosage is only one example to illustrate and is not limited to the above range.

In addition, the present invention 5-deoxycamphorol of the formula (1) can be used as an additive in pharmaceuticals or food. The composition according to the present invention may be a pharmaceutical composition or a food composition, the food composition in the present invention is intended to include food, dietary supplements, food additives, beverages and beverage additives and the like.

Preferably, the composition of the present invention may be a pharmaceutical composition comprising the active ingredient of the 5-deoxycamphorol of the formula (1).

That is, the method of the present invention, a method using the inhibitory effect of COX-2 expression of 5-deoxycamphorol, can be used to prevent and treat skin inflammation, skin disease and skin cancer.

The active ingredient having the pharmacological activity may be an anti-inflammatory agent, and more preferably, an agent for preventing and treating skin inflammation, skin disease and skin cancer.

In addition, when 5-deoxy camphorol is used as a food additive in the present invention can provide a food or beverage composition for health promotion. When the composition of the present invention is used as a food additive, various foods such as meat, cereals, caffeine drinks, general beverages, chocolate, breads, snacks, confectionery, pizza, ramen, other noodles, gums, ice creams, alcoholic beverages, Alcohol, vitamin complexes and other health supplements can be used, but is not limited thereto.

The 5-deoxycamphorol of the present invention may be prepared by adding a predetermined amount to the cooked food to prepare food or drink for health promotion, in which case the content of 5-deoxycampol in the finally prepared food or drink is 0.01 To 50% by weight. In this case, if the content of 5-deoxycampolol is less than 0.01% by weight, the effect may be insignificant, and when it exceeds 50% by weight, the effect increase compared to the amount of use is not economical.

As described above, 5-deoxycamphorol according to the present invention has an increased effect of inhibiting COX-2 expression by ultraviolet irradiation. Therefore, the composition to which 5-deoxycamphor is added is an excellent skin inflammation and skin disease prevention and treatment.

Hereinafter, preferred examples are provided to aid in understanding the present invention. However, these examples are only to illustrate the present invention more easily, the present invention is not limited to these.

Example 1 Determination of COX-2 Expression Inhibition Effect of 5-deoxycamphorol

Rat skin epithelial cells JB 6 P + cells (obtained from the American Cell Line Bank (ATCC)) contained 7.5 mg / L of penicillin and 7.5 mg / L of streptomycin in 5% fetal bovine serum (FBS) and MEM medium. Incubated in a 5% CO 2, 37 ° C. incubator (Forma Scientific Co., Marjetta, OH, USA) The media compositions were all used by GIBCO BRL (Grand Island, NY, USA).

Western blotting analysis by Upham, BL, Kang, KS, Cho, HY, & Trosko, JE Carcinogenesis to determine if 5-deoxycampol inhibits the expression of COX-2 by UVB. 18: 37-42, 1997].

5-deoxy camphorol was pretreated for 30 minutes by concentration, and then irradiated with UVB 0.05 J / cm ² and incubated for 4 hours. To extract the protein from the cultured cells were extracted with 20% SDS containing 1 mM phenylmethylsulfonylfluoride (PMSF). Protein content was determined using a DC assay kit [Bio-Rad Corp., Richmond, CA, USA]. About 15 μg of protein from each protein extract was separated by electrophoresis into 10% SDS-PAGE. After reacting with a COX-2 polyclonal antibody [Cayman Chemical, Ann Arbor, MI, USA], the reaction was detected using an ECL kit [Amersham, Life Science, Denver, USA].

1 shows the inhibitory effect of COX-2 of 5-deoxycamphorol, lane 1 is the untreated control, lane 2 is the UVB irradiation group, lane 3 is UVB and 5-deoxycampolol 10 μM treatment group, Lane 4 is UVB and 5-deoxycampolol 20 μM treated group, Lane 5 is UVB and 5-deoxycampolol 40 μM treated group. It was confirmed that the band of COX-2 decreased in a concentration dependent manner. Beta-actin proves that experiments with the same amount of cells.

FIG. 1B is a graph comparing the expression of COX-2 to the amount of beta actin in the results of FIG. 1A. The inhibitory effect of expression of COX-2 by 5-deoxycamphorol is enhanced in a concentration dependent manner.

Therefore, it can be seen that 5-deoxykaempferol decreases the concentration-dependent expression of increased COX-2 by ultraviolet light.

Example  2: 5- Deoxycamphor p38 , JNK ( Jun  N- terminal kinase ), Akt  Measurement of phosphorylation inhibitory effect

To determine whether 5-deoxycamphor inhibits phosphorylation of p38, JNK, and Akt, a major phenomenon that occurs when COX-2 expression is increased by UV light, Western blotting assay was used. The rolls were pretreated for 30 minutes by concentration, irradiated with UVB 0.05 J / cm ² , incubated for 30 minutes, and then extracted with protein. P38, JNK, Akt phosphorylation and non-phosphorylation of the selective antibody [Upstate Biotechnology, Inc., Lake Placid, NY] was used for the reaction.

Figure 2 shows the phosphorylation inhibitory effect of 5-deoxycamphor, lane 1 is a non-treated control, lane 2 is UVB irradiation group, lane 3 is UVB and 5-deoxycampolol 10 μM treated group, Lane 4 is UVB and 5-deoxycampolol 20 μM treated group, Lane 5 is UVB and 5-deoxycampolol 40 μM treated group. It was confirmed that the bands of p-p38, p-JNK, and p-Akt, which were phosphorylated by UVB, decreased in a concentration-dependent manner. Non-phosphorylated p38, JNK, and Akt proved to be tested with the same amount of cells. Therefore, it can be seen that 5-deoxykaempferol reduces the concentration-dependent expression of p38, JNK and Akt phosphorylated by UV light.

Example  3: 5- Deoxycamphor  Inflammatory Transcription Factors NF - kB  Activity inhibitory effect

NF-kB luciferase reporter plasmid, an inflammation-inducing transcription factor, was stably introduced into JB 6 P + cells to determine their activity. After pretreatment of 5-deoxycampolol for 30 minutes by concentration, UVB 0.05 J / cm ² was irradiated and incubated for 2 hours, Luciferase solution was added to measure the luminometer. As shown in FIG. 3, it can be seen that 5-deoxycampol inhibits the activity of UVB-induced NF-kB transcription factor. Therefore, it can be seen that 5-deoxykaempferol decreases the concentration-dependent activity of NF-kB transcription factor increased by ultraviolet rays.

Example  4: 5- Deoxycamphor In the rat skin COX -2 expression inhibitory effect

Female ICR mice were treated with 5-deoxycamphorol for 1 hour by concentration for 5 hours per group, 5 mice per group, and UVB was irradiated at 0.5 J / cm 2, and mice were sacrificed after 6 hours. The back skin of the mouse was removed, fat was removed, and protein was extracted. Using the COX-2 polyclonal antibody described above was confirmed by Western blotting analysis. Figure 4 shows the inhibitory effect of COX-2 of 5-deoxycamphorol in mice, lane 1 is untreated control, lane 2 UVB irradiation group, lane 3 UVB and 5-deoxy camphorol 50 μM Treatment group, lane 4 is UVB and 5-deoxycampolol 200 μM treatment group, lane 5 is UVB and 5-deoxycampolol 800 μM treatment group. It was confirmed that the band of COX-2 decreased in a concentration dependent manner. Beta-actin proves that experiments with the same amount of cells. Thus 5-deoxykaempferol is a mouse in In vivo experiments confirmed that the concentration-dependent inhibition of the expression of COX-2 increased by ultraviolet light.

Example 5: Inhibitory effect of skin cancer caused by UVB irradiation of 5-deoxycamphor

After 15 hours of SKH-1 hairless mice per group, a total of 60 mice were treated with 5-deoxycampolol three times a week for 1 hour, and then irradiated with UVB (0.18 J / cm ² ) for 30 weeks. Was carried out. Skin cancers were tabulated by counting skin cancers as more than 1 mm ² in diameter for more than two weeks.

FIG. 5 shows that 5-deoxycamphor inhibits skin cancer caused by UVB irradiation. The control group of FIG. 5A is acetone-treated group, and the UVB group is UVB-irradiated group after one hour pretreatment with acetone. The deoxy camphorol 40 and 100 μM groups were treated with 5-deoxycamphor at their respective concentrations. FIG. 5A shows that a representative mouse was taken at the end of the experiment, and photographs were taken. It was confirmed that the number of mouse skin cancers increased by UVB irradiation decreased depending on the concentration of 5-deoxy camphorol treated. 5b shows the number of mouse skin cancers observed for 30 weeks in a graph, and it was confirmed that the mouse skin cancers increased by UVB irradiation decreased concentration-dependently of treated 5-deoxycamphorol.

Example 6: Anticancer Agent Preparation

6-1. Powder

1 g of lactose was mixed with 2 g of 5-deoxycamphorol and filled into an airtight cloth to prepare a powder.

6-2. refine

Tablets were prepared by mixing 100 mg of 5-deoxycampolol, 100 mg of corn starch, 100 mg of lactose and 2 mg of magnesium stearate, followed by compression according to a conventional tablet preparation method.

6-3. Capsule

A capsule was prepared by mixing 5-deoxycampolol 100 mg, corn starch 100 mg, lactose 100 mg, and magnesium stearate 2 mg, and then filling the gelatin capsules.

6-4. Injection

Injectable drugs were prepared by adding an appropriate amount of distilled water for injection to 100 mg of 5-deoxycampolol, adjusting the pH to about 7.5, and then filling and sterilizing a 2 ml ampoule.

Example 7: Functional Food Preparation

7-1. Wire

Brown rice, barley, glutinous rice, and yulmu were alphatized by a known method, and then dried and roasted to prepare a powder having a particle size of 60 mesh using a grinder. Black beans, black sesame seeds and perilla were each steamed and dried in a known manner, then roasted and ground to prepare a powder having a particle size of 60 mesh.

Then, 30% by weight brown rice, 15% by weight of barley, 20% by weight of barley, 9% by weight of glutinous rice, 7% by weight of perilla, 8% by weight of black soybeans, 7% by weight of black sesame, 3% by weight of 5-deoxycampol, Ganoderma 0.5 A wire was prepared by mixing the wt% and 0.5 wt% of the turmeric.

7-2. Chewing gum

Chewing gum was prepared in a conventional manner by combining 20% by weight of gum base, 76.9% by weight of sugar, 1% by weight of perfume, 2% by weight of water, and 0.1% by weight of 5-deoxycampol.

7-3. candy

Candy was prepared by the conventional method by combining 60% by weight of sugar, 39.8% by weight of starch syrup, 0.1% by weight of perfume, and 0.1% by weight of 5-deoxycamphorol.

7-4. Biscuits

Force 1st class 25.59 wt%, 1st class gravity 22.22 wt%, white sugar 4.80 wt%, salt 0.73 wt%, glucose 0.78 wt%, palm shortening 11.78 wt%, ammonium 1.54 wt%, sodium bicarbonate 0.17 wt%, sodium bisulfite 0.16 wt %, Rice flour 1.45 wt%, Vitamin B₁0.0001 wt%, Vitamin B20.0001 wt%, Milk flavor 0.04 wt%, Water 20.6998 wt%, Whole milk powder 1.16 wt%, Substitute milk powder 0.29 wt%, Calcium phosphate 0.03 wt% , Biscuits were prepared in a conventional manner by combining 0.29 wt% of spraying salt, 7.27 wt% of spray oil, and 1 wt% of 5-deoxycampolol.

7-5. Health drink

0.26% by weight of honey, 0.0002% by weight of thioctoamide, 0.0004% by weight of nicotinic acid, 0.0001% by weight of riboflavin sodium hydrochloride, 0.0001% by weight of pyridoxine hydrochloride, 0.001% by weight of inositol, 0.002% by weight of orthoic acid, 98.7362% by weight of water and 5- 1 wt% of deoxycamphorol was blended to prepare a health beverage in a conventional manner.

7-6. sausage

Pork 65.18%, chicken 25%, starch 3.5%, soy protein 1.7%, salt 1.62%, glucose 0.5%, glycerin 1.5% and 1% by weight of 5-deoxycampol Sausage was prepared by the method.

7-7. Health Supplement

55% by weight of spirulina, 10% by weight of guar gum enzyme digestion, 0.01% by weight of vitamin B₁ hydrochloride, 0.01% by weight of vitamin B6 hydrochloride, 0.23% by weight of DL-methionine, 0.7% by weight of magnesium stearate, 22.2% by weight of lactose and 1.85% by weight of corn starch And 10% by weight of 5-deoxycamphorol to prepare a tablet-type health supplement in a conventional manner.

7-8. mainstream

0.5% of 5-deoxycampolol is mixed with soju, beer, liquor or fruit liquor to make an emulsion, and then centrifuged at 7,000 rpm for 15 minutes in vacuum or mixed at 9,000 rpm with a high speed mixer to produce 5-deoxycampolol. Liquor containing the mixture was prepared.

1 is a result of measuring the inhibitory effect of 5-deoxycamphor in the expression of cyclooxygenase-2 (COX-2) generated by ultraviolet irradiation in rat dermal epithelial cell line JB6 P +.

FIG. 2 shows the results of measuring the inhibitory effect of 5-deoxycamphor on phosphorylation of p38, JNK (Jun N-terminal kinase), and Akt, proteins that regulate COX-2 expression when UV-irradiated to JB6 P +. .

3 is a measurement result of the inhibitory effect of 5-deoxycamphor on the activation of NF-kB, an inflammation-inducing transcription factor.

4 shows the results of measuring the protective effect of 5-deoxycamphor on increased COX-2 expression when irradiated with ultraviolet light from mouse skin.

5 is a result of measuring the inhibitory effect of 5-deoxycamphor on increased skin cancer by irradiating the skin of the mouse with ultraviolet rays.

Claims (10)

A composition for preventing and treating skin inflammation and skin diseases comprising an effective amount of 5-deoxycamphorol of the following formula as an active ingredient.

The composition for preventing and treating skin inflammation and skin disease according to claim 1, wherein the composition comprises 0.1 wt% to 50 wt% of an active ingredient. The method of claim 1, wherein the effective amount is 10μM to 1mM for skin inflammation and skin disease prevention and treatment composition. 4. A composition according to any one of claims 1 to 3, wherein said composition is a medicament. The composition of claim 4, wherein the medicament is an anticancer agent. 6. The composition of claim 5, wherein the composition has an anticancer effect against skin cancer. The composition of claim 5, wherein the composition comprises 0.1 wt% to 50 wt% of an active ingredient. The composition of claim 5 wherein the effective amount is 10 μM to 1 mM. The composition of any one of claims 1 to 3, wherein the composition is a food, food additive, beverage, or beverage additive. 10. The composition of claim 9, wherein the food is gum, candy, biscuit, sausage, bread, or jelly.

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