KR100823537B1 - Compositions for Anti-obesity Comprising α-Bisabolol as an Active Ingredient - Google Patents

Abstract

A composition comprising alpha-bisabolol is provided to show excellent obesity inhibitory effect with safety and without side effects due to being derived from a natural material. An anti-obese composition comprises 0.0001-10 wt.% of alpha-bisabolol obtained by extracting Satureja punctata L. with water, C1-4 alcohol, a mixture solvent of water and C1-4 alcohol, acetone, ethyl acetate, chloroform or 1,3-butylene glycol at a temperature of 0-50 deg.C as an effective ingredient. The composition is a pharmaceutical composition.

Description

Composition for anti-obesity including alpha bisabolol as an active ingredient {Compositions for Anti-obesity Comprising α-Bisabolol as an Active Ingredient}

The present invention relates to a composition comprising alpha bisabolol as an active ingredient having excellent product stability and being safe to use without side effects on the skin and having excellent skin condition improvement effect, immunomodulatory effect and anti-obesity effect.

Human skin color is determined by the concentration and distribution of melanin inside the skin. Melanin pigment produced by melanocytes of human skin is a phenolic polymer having a complex form of black pigment and protein, and plays an important role in preventing skin damage caused by ultraviolet rays. It is reported that the action of tyrosinase in melanocytes is the most important for melanin biosynthesis, and tyrosinase is a type of amino acid tyrosine (DOPA) which is an intermediate of melanin polymer production. It also plays a key role in skin blackening by converting it to dopaquinone.

In general, skin wrinkles are known to be produced by a combination of factors, such as the skin's moisture content, collagen content, and the ability to act on the environment. Among these factors, the most influential effect on the formation of wrinkles is the expression and activity of collagenase, a collagen degrading enzyme that reduces collagen production and collagen content.

Hormonal imbalance is aggravating due to environmental pollution and automobile exhaust gas. As a result, the incidence of hair loss increases, the age of incidence decreases, and incompatibility of the skin immune system is caused, resulting in various skin diseases. In addition, the population suffering from obesity is rapidly increasing despite the age of children due to the changes in the diet of meat and meat.

Therefore, the development of materials that can effectively solve various phenomena, such as blemishes or freckles, wrinkles caused by ultraviolet rays, obesity, immune imbalance, hair loss, etc., which are not only aesthetical but also serious social problems, have been studied in depth.

Throughout this specification, numerous patent documents are referenced and their citations are indicated. The disclosures of the cited patent documents are incorporated by reference herein in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly described.

The present inventors have tried to develop a new single substance which is excellent in skin whitening, wrinkle improvement, skin damage by ultraviolet rays, prevention of hair loss or hair growth, anti-oxidation and anti-obesity effects, high stability, and no skin side effects. As a result, the present invention was completed by confirming that preparing a composition containing alpha bisabolol as an active ingredient can provide a composition for improving skin condition, antioxidant and anti-obesity having excellent efficacy and safety.

Accordingly, an object of the present invention is to provide a composition for improving skin conditions, wherein the skin condition is a skin condition improvement composition characterized in that the skin damage or hair growth by whitening, wrinkles, ultraviolet rays.

Another object of the present invention to provide an antioxidant composition.

Another object of the present invention to provide an anti-obesity composition.

Other objects and advantages of the present invention will become apparent from the following detailed description and claims.

According to an aspect of the present invention, the present invention is a composition for improving skin conditions comprising alpha bisabool (α-bisabolol) as an active ingredient, the skin condition is skin damage or hair growth caused by whitening, wrinkles, ultraviolet rays It provides a composition for improving skin condition, characterized in that.

According to another aspect of the present invention, the present invention provides an antioxidant composition comprising alpha bisabolol (α-bisabolol) as an active ingredient.

According to another aspect of the invention, the present invention provides an anti-obesity composition comprising alpha bisabolol (α-bisabolol) as an active ingredient.

The inventors of the present invention are excellent in product stability and can be used safely without side effects on the skin, skin whitening, wrinkle improvement, improvement of skin damage caused by ultraviolet light or promote hair growth, anti-obesity and antioxidant effect is an effective substance with natural plant Efforts have been made to develop the target. As a result, it was confirmed that the preparation of a composition containing alpha bisabolol as an active ingredient can provide a composition for improving skin condition, antioxidant and anti-obesity having excellent efficacy and safety.

Alpha bisabolol used as an active ingredient in the composition of the present invention is a monocyclic sesquiterpene alcohol as the main active ingredient of chamomile essence oil. Alpha bisabolol has the IUPAC name (2S) -6-methyl-2-[(1R) -4-methyl-1-cyclohex-3-enyl] hept-5-en-2-ol ((2S) -6- methyl-2-[(1R) -4 -methyl-1-cyclohex-3-enyl] hept-5-en-2-ol), which is a plant-derived compound having the structure of formula (1). Chamomile is traditionally known to have anti-inflammatory, sedative and pain relief effects. In addition, its main component, bisabolol, has been used in various skin care products as a colorless and contagious liquid having anti-inflammatory, wound healing and antibacterial effects in cosmetic formulations.

Formula 1

Alpha bisabolol used as an active ingredient in the composition of the present invention can be extracted from natural sources, for example chamomile, such as Satureja punctata L plant, or chemically synthesized. Specifically, the alpha bisabolol can be obtained by using various extraction methods known in the art, and preferably, (a) water, (b) 1-4 carbon atoms at an extraction temperature of 0 ° C. to 50 ° C. Anhydrous or hydrous lower alcohols (methanol, ethanol, propanol, butanol, etc.), (c) a mixed solvent of the lower alcohols with water, (d) acetone, (e) ethyl acetate, (f) chloroform or (g) 1 , 3-butylene glycol can be obtained as an extraction solvent.

If necessary, in order to obtain the alpha bisabolol of the present invention, after the extraction solvent treatment can be obtained through an additional purification process. For example, separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity), and the like through various purification methods performed additionally. Bisabolol can be obtained.

The composition of the present invention has a skin whitening use. As used herein, the term "whitening" refers to the action of improving skin troubles that prevent blackening of the skin caused by the deposition of melanin pigment.

In the skin whitening composition of the present invention, alpha bisabolol exhibits an excellent whitening effect of inhibiting melanin production by inhibiting the activity of intracellular tyrosinase compared to arbutin, which is used as a conventional skin whitening agent, and has high stability and time course. There is almost no change in the content of the component and there are almost no side effects such as skin irritation. This fact is clearly demonstrated in the following examples.

In addition, the composition for improving the skin condition of the present invention has a wrinkle improvement use. The composition of the present invention exhibits excellent antiwrinkle effect through molecular mechanisms such as inhibiting collagenase activity and promoting collagen synthesis, which is clearly described in the following examples. Wrinkle improvement, which is the use of the composition of the present invention, is understood to include conventional skin protection applications (prevention of wrinkles, removal of wrinkles, inhibition of skin aging, etc.).

In addition, the composition for improving skin conditions of the present invention has the effect of alleviating, improving, preventing or treating skin damage caused by ultraviolet rays.

The composition for improving skin condition of the present invention works very effectively to promote hair growth or prevent hair loss. As used herein, the term “anti-hair loss” or “promoting hair growth” is used in the same sense, which has the same meaning as another term wool or hair growth promotion used in the art.

Alpha bisabolol, an active ingredient of the present invention, exhibits an antioxidant effect through an excellent free radical removal effect.

Antioxidant composition of the present invention can be applied to various diseases, diseases or abnormal conditions that can be prevented or treated by inhibiting or removing the oxidized state. Antioxidant-related diseases that can be prevented or treated by the compositions of the invention include, but are not limited to, atherosclerosis, coronary artery disease, coronary artery restenosis, neurodegenerative diseases such as reperfusion injury, Parkinson's disease or Alzheimer's disease, stroke, cancer, aging , Cardiovascular disease, osteoporosis, central nervous system disorders, peripheral vascular disease, and dyspnea. Most preferably, the antioxidant composition of the present invention is used for the prevention or treatment of aging.

The correlation between many disease states and free radical damage is well established and many cellular components, including enzymes, ion channels, structural proteins and membrane lipids, are potential targets for reactive free radical species (Rice-Evans C). , Mol Aspects of Med 13 (1): 1-111 (1992). The reaction of these potential targets with free radicals impairs a range of cellular functions leading to pathological changes that ultimately kill cells. In addition, various diseases are caused by physiological oxidation, US Pat. No. 5,750,351; 5773209; 5773231 and 5846959.

The antioxidant composition of the present invention protects DNA, proteins (including lipoproteins) and membrane lipids from oxidative damage with an excellent free radical removal effect.

In addition, alpha bisabolol, which is an active ingredient of the composition of the present invention, has an excellent anti-obesity effect.

Alpha bisabobolol of the formula (1) of the present invention exhibits a skin whitening effect by inhibiting melanin production and / or tyrosinase activity in the derivative obtained by the addition or substitution reaction of a substituent commonly practiced in the art, It is clear to those skilled in the art in view of the state of the art to include wrinkle improvement, skin damage caused by ultraviolet light, hair growth or hair loss prevention, antioxidant and anti-obesity effects. More specifically, the alpha bisabolol used as an active ingredient in the present invention is not only the compound of Formula 1, but the structure of Formula 1 as a nucleus, and various substituents and substituent coupling reactions known in the art. Derivatives of the general formula (1) obtained are also included in the scope of the present invention. For example, even when a hydroxyl group, a halo group, a nitro group, or an alkyl group having 1 to 3 carbon atoms is bonded to Formula 1, it is determined to exhibit the same or similar efficacy as the alpha bisabolol of Formula 1, and such substituted derivatives may also be It is included in a range.

In a preferred embodiment of the present invention, the alpha bisabolol as the active ingredient is 0.00001 to 15.0% by weight, more preferably 0.0001 to 10% by weight, most preferably 0.0001 to 5% by weight based on the total composition. When the weight of the alpha bisabolol as an active ingredient is less than 0.00001% by weight, the effect is less likely to occur, and when it exceeds 15.0% by weight, the effect of increasing the content is very small and the stability of the formulation is not secured. There is this.

According to a preferred embodiment of the present invention, the composition of the present invention is a cosmetic composition.

The components included in the cosmetic composition of the present invention include components conventionally used in cosmetic compositions in addition to alpha bisabolol as an active ingredient, and include, for example, conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments and perfumes, And carriers. In addition, the cosmetic composition may further include a skin absorption promoting substance to enhance the effect.

The cosmetic composition of the present invention may be prepared in any formulation commonly prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing , Oils, powder foundations, emulsion foundations, wax foundations and sprays, and the like, but are not limited thereto. More specifically, it may be prepared in the form of a flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

When the formulation of the present invention is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. Can be.

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, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Soluble cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

The composition of the present invention may be prepared as a pharmaceutical composition.

When the composition of the present invention is made into a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. The pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives and the like in addition to the above components. Suitable pharmaceutically acceptable carriers and agents are Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, humans by various routes such as oral or parenteral, such as oral, rectal or intravenous, muscle, subcutaneous, intrauterine dura or cerebrovascular vessels. It can be administered by internal injection. It is preferably applied in the form of topical application during parenteral administration, more preferably by application.

Suitable dosages of the pharmaceutical compositions of the present invention may vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be. The dosage of the pharmaceutical composition of the present invention can be administered once or several times a day in an oral dosage form of 0.1-100 mg / kg on an adult basis. It is recommended to apply 1 to 5 times a day in an amount of 3.0 ml to continue for 1 month or more. However, the dosage does not limit the scope of the present invention.

The pharmaceutical compositions of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. The formulation may be used in any form suitable for pharmaceutical preparations, including powders, granules, tablets, capsules, suspensions, emulsions, syrups, oral formulations such as aerosols, external preparations such as ointments, creams, suppositories, and sterile injectable solutions. It may further comprise a dispersant or stabilizer.

In addition, the composition of the present invention may be prepared as a food composition.

When the composition of the present invention is made of a food composition, it contains not only alpha bisabool as an active ingredient, but also components commonly added in food production, and include, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavors. Include the first. Examples of the above carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As flavoring agents, natural flavoring agents (tauumatin, stevia extract (for example rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

 For example, when the food composition of the present invention is prepared with a drink, the citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, tofu extract, jujube extract, licorice extract, etc. Can be.

On the other hand, skin specific stimulation test for alpha bisabolol in a specific embodiment of the present invention it was found that alpha bisabool is a natural material harmless to the human body. Therefore, the alpha bisabolol of the present invention has little toxicity and side effects, so it can be used safely even in long-term use, and in particular, it can be safely applied to cosmetics, pharmaceutical and food compositions as described above.

The features and advantages of the present invention are summarized as follows:

(Iii) The composition of the present invention contains alpha bisabolol as an active ingredient.

(Ii) Alpha bisabolol, an active ingredient, inhibits the activity of intracellular tyrosinase by inhibiting the activity of melanin by inhibiting the activity of intracellular tyrosinase, compared to arbutin, which is used as a conventional skin whitening agent, and inhibiting collagenase activity and promoting collagen synthesis. Through the molecular mechanisms, such as excellent wrinkle improvement effect, the effect of improving the skin damage by ultraviolet light and has an excellent skin condition improvement effect that works very effectively in promoting hair growth or preventing hair loss.

(Iii) Alpha bisabolol, which is an active ingredient, also has excellent anti-obesity and antioxidant effects.

(Iii) The composition of the present invention can be safely applied to cosmetics, pharmaceuticals and food compositions without cytotoxicity and skin side effects.

As described in detail above, the present invention relates to a composition for improving skin condition comprising alpha bisabolol (α-bisabolol) as an active ingredient. Alpha bisabolol, an active ingredient used in the composition of the present invention, inhibits the activity of intracellular tyrosinase and inhibits melanin production, inhibits collagenase activity, and inhibits the activity of intracellular tyrosinase compared to arbutin, which is used as a conventional skin whitening agent. Molecular mechanisms such as promoting collagen synthesis have excellent anti-wrinkle effect, improved skin damage caused by UV light, and excellent skin condition which is very effective in promoting hair growth or preventing hair loss. Has In addition, there is no cytotoxicity and skin side effects, so it can be safely applied to cosmetics, pharmaceutical and food compositions.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, and the scope of the present invention is not limited by these examples in accordance with the gist of the present invention to those skilled in the art. Will be self explanatory.

Example  1: alpha In Bisaboall  Melanin production inhibitory effect

Rat B16 melanoma cells (Korea Cell Line Bank) were used to measure the inhibitory effect of melanin production by alpha bisabolol, and compared with the melanin production inhibitory effect by arbutin known to inhibit melanin production.

In this experiment, murine melanoma (B-16 F10) cells (Korea Cell Line Bank) were used in DMEM medium containing 10% fetal bovine serum (FBS) in a 6-well plate at 1 × 10 ⁵ per well. After inoculation with dogs, the cells were incubated at 5% CO ₂ and 37 ° C until at least about 80% were attached to the bottom of the well. After incubation, the medium was removed and the sample was replaced with medium diluted to an appropriate concentration and incubated at 5% CO ₂ , 37 ° C for 3 days. The concentration range of alpha bisabolol was determined to be 1 ppm, 5 ppm and 10 ppm without cytotoxicity. The cells from which the medium was removed were washed with PBS (phosphate buffer saline) and treated with trypsin to recover the cells. The recovered cells were measured by using a hematocytometer (Tiefe Depth Profondeur 0.100 mm, Paul Marienfeld GmbH & Co. KG, Germany), and then centrifuged at 5,000 to 10,000 rpm for 10 minutes. It was removed to obtain pellets. The cell pellet was dried at 60 ° C., and then 100 μl of 1 M sodium hydroxide solution containing 10% DMSO was added to obtain intracellular melanin in a 60 ° C. thermostat. Using this solution, the absorbance was measured at 490 nm using a microplate reader (Bio-Tek ELx8081U, USA) to determine the amount of melanin per cell. The results are shown in Table 1 below.

sample Treatment Concentration (ppm) Melanin production inhibition rate (%) Arbutin 5 22 Alpha bisabolol One 24 Alpha bisabolol 5 36 Alpha bisabolol 10 49

As shown in Table 1, alpha bisabolol has a higher melanin production inhibition rate even when the treatment concentration is lower than arbutin, and it can be seen that the melanin production inhibition rate increases as the treatment concentration of alpha bisabolol is increased.

Example  2: alpha In Bisaboall  by Intracellular Tyrosinase  Activity Inhibition Effect Measurement

Rat B16 melanoma cells were used to measure the inhibitory effect of melanin production by alpha bisabolol and compared with the inhibitory effect of intracellular tyrosinase activity by arbutin known to inhibit melanogenesis.

This experiment was performed by inoculating murine melanoma (B-16 F1) cells in 6-well plates at 1 × 10 ⁵ per well in DMEM medium containing 10% FBS (fetal bovine serum). Under% CO ₂ , 37 ° C., the cells were incubated until at least about 80% adhered to the bottom of the well. After incubation, the medium was removed, the sample was replaced with a medium diluted to an appropriate concentration, and then incubated for 3 days under 5% CO ₂ , 37 ° C. The concentration range of alpha bisabolol was determined to be 1 ppm, 5 ppm and 10 ppm without cytotoxicity. The cells from which the medium was removed were washed with PBS (phosphate buffer saline) and treated with trypsin to recover the cells. The recovered cells were measured by using a hematocytometer, and then centrifuged at 5,000 to 10,000 rpm for 10 minutes, and then the supernatant was removed to obtain pellets. The cell pellet was pulverized using lysis buffer, and then centrifuged at 12,000 rpm for 10 minutes to collect supernatant. Using this solution, absorbance at 492 nm was measured with a microplate reader to determine cell-specific allowance tyrosinase activity. The results are shown in Table 2 below.

sample Treatment Concentration (ppm) Inhibition rate of intracellular tyrosinase activity (%) Arbutin 5 45 Alpha bisabolol One 13 Alpha bisabolol 5 38 Alpha bisabolol 10 58

As shown in Table 2, it was found that alpha bisabolol had higher intracellular tyrosinase inhibition rate than arbutin. It was also found that the degree of inhibition of tyrosinase activity of alpha bisabolol is concentration-dependent.

Example  3: Evaluation of the whitening effect at the animal level

Similar to humans, the whitening effect of alpha bisabolol was measured using brown malmoto (Tortoiseshell guinea pigs; Brown River Guinea pigs, Charles River Laboratories, Inc.), which is known to cause pigmentation by ultraviolet rays.

In order to cause pigmentation due to ultraviolet (UV) in the brown molemoto, a light shielding aluminum foil having a square window of 3 × 3 cm ² is adhered to the skin from which the hair of the brown molemoto coordination is removed, and then SE lamp ( Ultraviolet rays were irradiated with a wavelength of 290-320 nm (Toshiba) (total amount of irradiation energy = 1350 mJ / cm ² ). After UV irradiation, the aluminum foil was peeled off and the samples (alpha bisabolol and arbutin) were applied in the following manner. Pigmentation appeared 2-3 days after UV irradiation and reached a peak after about 2 weeks, from which each sample was applied.

The number of coatings was continued once or twice a day for 50 days. The sample was dissolved and diluted using a specific solvent (propylene solvent: ethanol: water = 5: 3: 2: mixed solvent), and applied with a cotton swab, and prepared a control site for necessarily applying the solvent to other sites. Along with the determination of effects, the cumulative stimulus was also observed.

The effect was determined by measuring the degree of black and white of the skin using a color difference meter (CR2002 chromameter, Minolta, Japan), the results are shown in Table 3 below. L ^* A ^* B ^* color system is used to display color, and L ^* value was used as an index in this invention. L ^* value was calibrated with a standard whiteboard, and L ^* value was measured 5 times or more at one site, and pigmentation was equalized. The difference (ΔL ^* ) of the skin color at the start of application and at the end of application was determined and the effect was determined using this value. The results are shown in Table 3 below.

△ L ^* = L ^* value after 00 days application - L ^* value of the coating start date

Comparing the ΔL ^* value with respect to the sample application site and the control site, the effect of the whitening material can be seen.

sample Treatment Concentration (%) Whitening effect (△ L) Alpha bisabolol 0.2 0.42 Alpha bisabolol 1.0 0.67 Arbutin 1.0 0.54 Control - 0.35

As shown in Table 3, alpha bisabolol showed a whitening effect in a concentration-dependent manner. In addition, alpha bisabolol showed better whitening effect than arbutin, and no cumulative irritation was observed during the test application of the test substance, and thus safety was also excellent.

Example  4: alpha Bisabolol  Safety confirmation experiment on human skin

Example  4-1: Alpha Bissabolol  inclusive External skin preparations  Produce

In order to confirm that alpha bisabolol, which has been found to be excellent in whitening effect as described above, is safe for human skin, an external skin preparation containing alpha bisabolol and arbutin was prepared and skin safety verification experiments were performed.

Skin external preparations containing alpha bisabolol and arbutin were prepared using the ingredients listed in Table 4 below. In Table 4, the control group is an external skin preparation that does not include a tyrosinase inhibitor, and test group 1 and test group 2 are external skin preparations including alpha bisabolol and arbutin, respectively.

For the preparation of external skin preparations, purified water, serine and butylene glycol were mixed and dissolved at 70 ° C. (aqueous part), and the other components except the three components and trimethanolamine were dissolved at 70 ° C. (oil phase part). The oil part was added to the water part and stirred with a homomixer (Tokushu Kika, Japan) to emulsify first, to which trimethanolamine was finally added. After removing the bubbles generated in the mixed solution, and cooled to room temperature to prepare a skin external preparation.

ingredient content (%) Control Test group 1 Test group 2 Purified water 72 72 72 glycerin 8.0 8.0 8.0 Butylene glycol 4.0 4.0 4.0 Alpha bisabolol 0 1.0 0 Arbutin 0 0 1.0 Caprylic Capri Triglyceride 8.0 8.0 8.0 Squalane 5.0 5.0 5.0 Cetearyl Glucoside 1.5 1.5 1.5 Sorbitan stearate 0.4 0.4 0.4 Cetearyl Alcohol 1.0 1.0 1.0 Trimethanolamine 0.1 0.1 0.1 Gross weight 100 100 100

Example  4-2: Accumulated skin irritation test

Whether or not alpha bisabolol irritates the skin by subjecting the upper forearm to a total of nine 24-hour cumulative blisters every other day for 30 healthy adults using each skin preparation prepared in Example 4-1. Whether or not was measured.

The patch method used the fin chamber (Finn chamber, Epitest Ltd, Finland). 15 µl of each of the external skin preparations was added dropwise to the chamber, followed by patching. The degree of response to the skin each time was scored using Equation 2 below, and the results are shown in Table 5 below.

Average responsiveness = [{(response x responsiveness) / (total number of subjects x highest score (4 points))} x 100] ÷ number of tests (9 times)

At this time, ± 1 point, + 2 points, + + 4 points in the reaction degree, and the average response was determined to be a safe composition when less than 3.

Test substance Number of subjects with response Average reactivity 1 week 2 weeks 3 weeks Primary Secondary 3rd 4th 5th 6th 7th 8th 9th ± + + + ± + + + ± + + + ± + + + ± + + + ± + + + ± + + + ± + + + ± + + + Control 2 - - --- --- --- --- --- --- --- --- 0.18 Test group 1 3-- One - - --- --- --- --- --- --- --- 0.37 Test group 2 3-- One - - --- --- --- --- --- --- --- 0.37 Inspector 30 30 30 30 30 30 30 30 30

In Test Table 1 and 2 in Table 5, the number of people corresponding to ±, + and ++ was 3, 0 and 1, respectively, and the rest did not appear. When calculated according to the formula described above, the average reactivity was 0.37, which was 3 or less, which was judged to be a safe composition.

As shown in Table 5, the external preparation for skin containing alpha bisabolol (test group 1) did not show a distinct cumulative stimulation pattern as the external preparation for skin including control or arbutin and was determined to be safe for human skin.

Example  5: antioxidant effect

Superoxide dismutase (SOD) is known as an antioxidant that converts superoxide anions into H ₂ O ₂ and O ₂ . This experiment is a method of evaluating the antioxidant capacity of a sample by observing the rate at which the superoxide anion generated by Xanthine oxidase was removed by the sample used in the test. The experiment was carried out using a kit purchased from Dojindo (Japan) (SOD Assay Kit-WST), and was tested according to the manufacturer's protocol. In brief, the effect was determined by observing the removal rate of superoxide anion by the sample, the presence or absence of antioxidant function. The experimental results are summarized in Table 5.

sample Treatment Concentration (ppm) Superoxide Anion Removal Rate (%) Alpha bisabolol One 8 Alpha bisabolol 5 18 Alpha bisabolol 10 34

As shown in Table 6, it can be seen that the alpha bisabolol of the present invention has an antioxidant effect in a concentration-dependent manner.

Example 6: Anti-inflammatory efficacy evaluation

In order to examine the anti-inflammatory effect, COX activity inhibitory activity was performed according to a conventional method in THP-1 cells, which are human monocytes. COX activity was measured by the Method FJ Van de Ouderaaa and Muytenhek announced in This was done according to Enzymology 43: 9 (1994). THP-1 cell line (Korea Cell Line Bank) was cultured and the cells were divided into 24-well plates. The incubation volume per well was adjusted to 500 μl, and 2 μl of sample dissolved in LPS (lipopolysaccharide, Sigma) (1 μg / ml) and the solvents shown in the following table were added, and then cultured under the same conditions for 24-48 hours. After 24-48 hours, each well was added with calcium ionophore (Sigma) and 1 μl of [1-14C] arachidonic acid (in EtOH, 0.1 μCi / ml, Sigma) and incubated for 10 minutes under the same conditions. After incubation, citric acid was added to each well, and the pH was adjusted to 3.5. Then, 500 μl of each culture solution was aliquoted into a microcentrifuge tube, and 700 μl of ethyl acetate was added thereto, followed by shaking extraction for 10 minutes. 500 µl of ethyl acetate layer was taken and concentrated for 20 minutes using a speed vacuum dryer. The concentrated residue was dissolved in 20 µl of ethyl acetate and applied to an TLC plate with an authentic standard. Radioactivity bands were identified by authentic eicosanoid standards, and the radioactivity of the identified bands was measured using a BAS 2000 bio-imaging analyzer (Fuji, Japan) (Table 7). .

sample COX activity (%) LPS (1 μg / ml) 100 LPS (1 μg / ml) + alpha bisabolol (1 ppm) 95 LPS (1 μg / ml) + alpha bisabolol (5 ppm) 75 LPS (1 μg / ml) + alpha bisabolol (10 ppm) 67

 As described in Table 7, it can be seen that the alpha bisabolol of the present invention effectively inhibits COX activity as the treatment concentration increases. It can be seen that there is an inflammation inhibitory effect.

Example 7: Immunosuppressive ability  evaluation

Whether the samples used in the present invention inhibit the immune related response related to inflammation was examined again through an interleukin-2 luciferase reporter activity test. The interleukin-2 promoter is reported to play an important role in the production of immune-related cytokines associated with inflammation. The following method was used to measure interleukin-2 luciferase activity. After dispensing 1 × 10 ⁶ cells into 6 wells of Jurkat cells (Korea Cell Line Bank), which is a human T lymphocyte cell line, interleukin-2 lucifera using a superperfect transfection reagent (In vitrogen) was used. Aze reporter plasmid DNA (IL-2 luciferase reporter plasmid DNA) was transfected. After 24 hours after transfection, PHA (Phytohemaglutinin, Sigma) (100 ng / ml) was treated to activate Jurkat cells and each sample was treated by concentration. After 24 hours, cells were collected and luciferase activity was measured using a luminometer (Berthold Technologies GmbH & Co.KG, Germany). The results are as follows.

sample Interleukin-2 Luciferase Activity PHA (100 ng / ml) 100 PHA (100 ng / ml) + alpha bisabolol (1 ppm) 92 PHA (100 ng / ml) + alpha bisabolol (5 ppm) 72 PHA (100 ng / ml) + alpha bisabolol (10 ppm) 58

As shown in Table 8, it can be seen that alpha bisabolol has an immunomodulatory function by inhibiting the expression of interleukin-2 as the treatment concentration increases.

Example 8: Wrinkle improvement effect measurement

The anti-wrinkle effect can usually be measured as collagen biosynthesis and collagenase degradation inhibitory effect and clinical trial in humans.

Human normal fibroblasts (Pacific) were inoculated into 6-well microplates containing DMEM medium (2 × 10 ⁵ cells / well) in a 2% CO ₂ incubator at 37 ° C. Time incubation. After 24 hours, the medium was removed from each well, samples were treated by concentration and then incubated again for 24 hours. After 24 hours, cell media were collected to prepare samples.

Collagen synthesis amount was measured by using the collagen measurement kit (Procollagen Type I C-peptide EIA kit (MK101), Takara, Kyoto, Japan) according to the manufacturer's protocol. The amount of Type I C-peptide (PICP) was measured and analyzed.

An antibody against collagenase was used as a method for measuring the activity of collagenase, an enzyme that degrades collagen. Collagenase activity was measured using a type I collagenase assay kit (Amersham Biosciences, RPN2629) and absorbance was measured with an ELISA reader (Bio-Tek ELx808 ™ Series Ultra Microplate Reader, UK). The measured standard values were expressed in the form of mean ± standard deviation, and the significance was tested by t-test using the SPSS / PC + program. The results are shown in Table 9 below.

Test Items Alpha Bisabolol (1ppm) Alpha Bisabolol (5ppm) Alpha Bisabolol (10ppm) Collagen Synthesis Growth Rate 23 43 76 Collagenase inhibition rate 12 28 43

As shown in Table 9, alpha bisabolol increased collagen synthesis in a concentration-dependent manner and also inhibited collagenase activity in a concentration-dependent manner. Therefore, it can be seen that alpha bisabolol has an antiwrinkle effect.

Example  9: Anti-obesity effect

Obesity inhibition test was generally carried out by the following method using a well-known animal. The results are shown in Table 10 below.

The method for measuring obesity inhibitory activity is as follows:

Crj: After 7 weeks of age, ICR male mice (Chicks River Co., Ltd.) were preliminarily bred for one week, and then used in experiments in seven groups. Animals are kept in a constant temperature room set at a temperature of 23 ± 1 ℃, a humidity of 55 ± 5% and an illumination time of 12 hours / day. It was made. Alpha bisabolol was prepared in liposome formulation (dissolved in 5% lecithin) to 0.1%, 1%. Each sample solution was adjusted to 0.1 ml per 10 g of mouse, and the dose was set at 1.5 g / kg and 1 g / kg. In addition, the control group was a 5% lecithin emulsion. Mice were fasted from the day before administration and were forcibly administered once the next day. The test period was two weeks, and body weight and general symptoms were measured and observed.

solvent Days Weight (g) No treatment group Alpha Bisabolol (0.1%) Alpha Bisabolol (1%) Liposomal Formulations 5 24 24 23 10 28 24 24 15 34 26 26

As can be seen in Table 10, alpha bisabolol showed an anti-obesity effect. It was observed that the higher the dosage concentration, the better the efficacy.

Example  10: hair loss prevention and Hair growth effect

In order to measure the effects on the prevention of hair loss and hair growth, a test was conducted using a hydrogel base containing only preservatives and thickeners. Using the prepared hair-promoting external preparation, it was applied to hair loss sites of 10 alopecia patients due to weakening or radiation exposure twice a day for 3 months each for 3 months. As a result, it was found that there were excellent effects such as the development of new hair roots in eight hair loss patients. The test results are as follows. As a control, moxidil sold in Hanmi Pharm was used.

No treatment group Moxidil Alpha bisabolol - ++ ++

As can be seen in Table 11, it can be seen that the alpha bisabolol of the present invention exhibits a hair regrowth effect similar to that of moxidil sold as a hair regrowth agent.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (3)

Anti-obesity composition comprising alpha bisabool (α-bisabolol) as an active ingredient. The composition of claim 1, wherein the alpha bisabolol is included in 0.0001-10% by weight based on the total weight of the composition. The composition of claim 1 or 2, wherein the composition is a pharmaceutical composition.