KR20200011781A - Cosmetic composition for improving the health of scalp comprising natural caffeine and chlorogenic acid - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving scalp health for hair loss prevention and hair growth, comprising, as main ingredients, natural caffeine and chlorogenic acid, which are main bioactive compounds in green coffee beans. Using coffee bean extracts as cosmetic compositions requires a large quantity of such coffee bean extracts in order to exhibit effective functionalities of caffeine and chlorogenic acid, and also, the coffee bean extracts are not preferably used as cosmetic compositions due to its aroma and color. According to the present invention, caffeine is isolated and purified from green coffee beans, and then mixed in a predetermined ratio with a natural substance in which chlorogenic acid is concentrated, and thus can produce, even in a small amount, a desired functional cosmetic composition. The mixture does not have toxicity to cells in the human body, and is excellent in terms of functions relating to skin health, such as antioxidant activity, skin whitening effect, and hair growth promoting effect. Therefore, the composite composition of caffeine and chlorogenic acid of the present invention can be beneficially used in cosmetic compositions for skin and scalp health effective to enhance the antioxidant function of the skin and to prevent hair loss.

Description

Cosmetic composition for improving the health of scalp comprising natural caffeine and chlorogenic acid}

The present invention relates to a cosmetic composition for improving scalp health containing natural caffeine and chlorogenic acid as an active ingredient.

In general, coffee belongs to the Rubiaceae family, first grown in Ethiopia in the 5th century, and began to spread to Islam and other countries by the 15th century, and is now one of the most consumed beverages in the world. I became one. Coffee is mainly grown by Coffea arabica and Canepora. Green bean refers to seeds whose outer skin, pulp, perchment and silver skin have been stripped from the fruit of the coffee tree, which is roasted by heating the coffee beans. It is called coffee roasted bean. After the roasting process, the coffee beans have a unique aroma and taste.

The development of functional cosmetics using caffeine, a well known functional material of coffee, is highly marketable and accessible to consumers. Chlorogenic acid (CGA), an antioxidant in coffee, is destroyed and destroyed during the production of coffee beans. Overseas diet foods using green soybean chlorogenic acid are sold, but it is known to cause problems when overdose. However, it is difficult for domestic companies to use natural purified chlorogenic acid and caffeine due to the high cost of imported green beans, lack of purified purity and bioactivity. As cosmetic materials, cosmetics utilizing expensive natural purified chlorogenic acid, quinic acid, capic acid (caffeic acid) and caffeine have surprisingly little technical development and products.

The molecular weight of caffeine is 194 MW, which is one of the purine bases contained in tea, coffee, black tea and the like. Cardiac agent, alkaloid, adenosine receptor inhibitory activity, taanodine receptor / calcium release channel opening promoting effect, cAMP-increasing effect by phosphodiester hydrolase inhibition (Schmidt et al., 2006; Nehlig et al., 1992 ).

Caffeine increases the secretion of neurotransmitters such as dopamine by interfering with the effects of adenosine, which inhibits the secretion of neurotransmitters. In addition to excitement, central nervous excitability, drowsiness prevention, skeletal muscle contraction, gastric juice, hyperactivity, cardiac, smooth muscle relaxation, diuretic, bowel movement, Parkinson's dementia treatment (Snel and Lorist 2011). Caffeine is an active substance in major foods such as coffee, cocoa and green tea. Caffeine plays a role in improving mood when drinking. Pure purified caffeine is used to treat obesity, dementia, and depression, and has been reported to prevent hair loss in recent years (Fischer et al., 2007; Otberg et al., 2007).

Chlorogenic acid is a natural compound composed of ester bonds of caffeic acid and quinic acid. It is known as an antioxidant due to its antioxidant effect and is known to be added to anti-diabetic functional foods by delaying the release of blood glucose after meals (Crozier et al., 2010; Del Rio et al., 2010). It is known that chlorogenic acid has a diet effect, but skin cell activity or keratinocyte, fibroblast, hair follicle cells, etc., skin wrinkle improvement and whitening effect that can be used in cosmetic technology development Research is required.

The catechol-type antioxidants that are not flavonoids include chlorogenic acid, caffeic acid and dihydrocaffeic acid (DHCA), and their use as a cosmetic material is important. Anthocyanins, which are flavone-based antioxidants, may be inadequate for cosmetics due to the nature of the pigment, so the utilization of quinone-based materials is very important. In Korea, there are very few research cases in which coffee-derived antioxidants, chlorogenic acid and caffeic acid, were developed as the main ingredients of cosmetics. With the development of the coffee industry, caffeine and chlorogenic acid are used as diet foods and additives, and hair loss prevention shampoos containing caffeine are imported and marketed.

However, there are no cosmetics with two main components, caffeine and chlorogenic acid, and for domestic products, caffeine and chlorogenic acid are not used as main bioactive substances. Commercially available products are coffee extracts that contain coffee extracts or cosmetics containing coffee polyphenols.

Body cosmetics and shampoos that simply contain 'coffee extracts' are sold in the category of 'coffee cosmetics', but there are very few cosmetic products made from pure purified natural caffeine or natural chlorogenic acid.

Therefore, in the present invention, the main ingredient is a mixture of caffeine and chlorogenic acid purified separately from green coffee beans, whose content ratio is adjusted, and a cosmetic composition for whitening effect, hair loss prevention, and scalp improvement.

As shown in FIG. 3, Benign prostatic hyperplasia (BPH) is a representative disease that occurs in the prostate gradually begins after the 40s, 60-70% occurs in the 60s and almost all males by 70 years old It is a very common disease. As age increases, the amount of male hormone produced by testicles in males decreases, but the activity of male hormone converting enzyme 5-α reductase increases, leading to the increase in the activity of active male hormones such as dihydrotestosterone (DHT). Increased amounts ultimately lead to an enlarged prostate.

In normal people, when the DHT increases due to excessive 5-α reductase activity, the androgen receptor binds to the androgen effect and ultimately hair loss progresses. Inhibition of 5-α reductase activity is an important means for the treatment of hair loss.

Hair is a skin organ that remodels itself during the circulatory cycles of the active, anagen, catagen, and telogen (Ahn et al., 2012). Under the influence of the androgen hormone, hair follicle miniaturization proceeds, resulting in short cycle growth (Whiting, 2001). Hair follicle size and growth length determine hair length and size, respectively (Cotsarelis and Millar, 2001). The normal growth phase lasts on average about 2-6 years and then has a short, transient transition in which cell death of hair follicles occurs (Otberg et la., 2007). Therefore, it is important for androgenetic alopecia (AGA) to prevent the miniaturization of hair follicles and prevent cell death to prolong the possible growth phase (Cotsarelis and Millar, 2001).

Other factors affecting male hair loss include highly reactive free radical molecules with unshared electrons that damage cells. This damages cell membranes, DNA, proteins, etc., leads to aging of the hair and reduced hair development (Wood et al., 2009). In particular, lipid peroxides accumulated in hair follicles cause hair loss early in the mouse's hair cycle (Naito et al., 2008). Therefore, the application of antioxidants is a good way to prolong hair growth and prevent hair loss.

In the present invention, a cosmetic composition for preventing and preventing alopecia of male alopecia by aging by utilizing the function of cell division and aging delay and the high antioxidant function of chlorogenic acid by the function of improving blood circulation and energy activity of caffeine using coffee as a material Developed.

Republic of Korea Patent Publication No. 10-2014-0058923 (published May 15, 2014)

Therefore, an object of the present invention was to purify the natural caffeine extracted from the coffee beans, to prepare an extract concentrated in chlorogenic acid from the coffee beans, evaluate the physiological activity of these substances on the human body and ultimately containing them as an active ingredient To provide.

One aspect of the present invention is to separate and purify the caffeine from the coffee beans, and then concentrated in the process to prepare a cosmetic composition material chlorogenic acid. Subsequently, antioxidant properties, human cell toxicity evaluation, human skin keratinocyte proliferation effect, keratin production ability, and hair growth function were evaluated. Accordingly, the present invention provides a cosmetic composition containing purified caffeine and chlorogenic acid alone or in combination based on the results of physiological activity evaluation.

In the present invention, natural caffeine was isolated and purified from green coffee beans, and then an extract of concentrated chlorogenic acid was separated from the fraction material. At least 10% concentrates of purified natural purified caffeine and chlorogenic acid, respectively, were prepared from the same coffee beans. Thereafter, a composite material was prepared in which these were mixed at an appropriate ratio. The composite material has both a physiological activity of caffeine and a chlorogenic acid, and shows a synergistic effect in the cosmetic composition. Generally, the caffeine concentration of coffee is about 1 to 3%, and the content of chlorogenic acid is about 0.1% or more and less than 1%. Therefore, when using bean extract, a large amount of bean extract should be used to show the effective physiological activity of caffeine. Likewise, in order to obtain the physiological activity of chlorogenic acid, a large amount of extract must be included in the cosmetic composition, thereby making cosmetic preparation impossible. In addition, there are many cosmetic compositions containing the original coffee bean extract, but containing a large amount of coffee extract in order to have a unique physiological activity of the coffee is difficult to use as a cosmetic because the color and aroma of the cosmetic changes. Therefore, in the present invention, caffeine was isolated and purified from the same coffee green beans, and coffee chlorogenic acid was prepared from the residue. According to the purpose, the composition of the two representative bioactive ingredients of coffee can be adjusted to maximize the bioactive functionality.

According to the present invention, a complex material in which a concentrate of caffeine and chlorogenic acid separated and purified from coffee beans, respectively, is mixed in a certain ratio and is not toxic to human cells and is useful for skin health such as antioxidant, whitening, and hair growth promotion. Therefore, the complex composition of caffeine and chlorogenic acid of the present invention can be usefully applied to the skin and scalp health cosmetic composition having the effect of enhancing the antioxidant function of the skin and preventing hair loss.

1 is a chemical structure of antioxidant caffeine, dihydrocaffeic acid (DHCA), caffeic acid (CA), and chlorogenic acid (CGA) of major catechols derived from green beans. It is shown.
Figure 2 shows the separation, purification and recrystallization of caffeine from coffee (left), the caffeine crystals of purity 99.95% obtained from caffeine recrystallization confirmed by HPLC (middle), 99.5% or more purified caffeine powder used in the present invention And a 50% purified chlorogenic acid powder picture (right).
Figure 3 relates to the control of the growth cycle of the hair by the administration of the complex and additives of natural caffeine and chlorogenic acid used in the present invention, the material shows a conceptual diagram to prevent hair loss and increase hair thickness by inducing and extending the growth phase will be. (A) is a conceptual diagram of androgenetic alopecia and prostate hyperplasia due to the increase of dihydrotestosterone (DHT) by 5-α reductase, (B) is scalp health and hair loss prevention by natural caffeine and chlorogenic acid of the present invention It shows a conceptual diagram.
4 is a flowchart illustrating a process of extracting caffeine and chlorogenic acid from hot coffee beans using hot water and methylene chloride solvent.
Figure 5 shows the evaluation of antioxidant activity by DPPH radical removal using caffeine, chlorogenic acid, coffee extract (Results are the means ± S. D of three samples, * p <0.05 vs. positive control, ** p <0.005 vs. positive control).
Figure 6 shows the evaluation of the antioxidant activity by ABTS · ⁺ radical scavenging activity using caffeine, chlorogenic acid, coffee extract (Results are the means ± S. D of three samples, * p <0.05 vs. positive control, ** p <0.005 vs. positive control).
Figure 7 shows the tyrosinase inhibition evaluation of caffeine, chlorogenic acid, coffee extract. This is an assessment of whitening functionality (Results are the means ± S. D of three samples, * p <0.05 vs. positive control, ** p <0.005 vs. positive control).
Figure 8 shows the evaluation on the human cytotoxicity and growth rate using caffeine, chlorogenic acid, coffee extract (Results are the means ± S. D of three samples, * p <0.05 vs. positive control, ** p <0.005 vs positive control).
9 shows the hair growth evaluation results of the mouse. Hair growth in the anagen phase of hair follicles was measured and evaluated by assigning hair growth scores from 1 to 5 points.

Thus, the present invention provides a cosmetic composition for improving scalp health containing natural caffeine and chlorogenic acid as an active ingredient.

The natural caffeine and chlorogenic acid may be included in 0.0001 to 50 parts by weight based on 100 parts by weight of the total composition, but is not limited thereto.

The cosmetic composition is a shampoo, hair treatment, hair tonic, flexible lotion, gel, water soluble liquid, milk lotion, cream, essence, skin toner, oil-in-water emulsion, water-in-oil It may be formulated as a formulation selected from the group consisting of emulsion, cleansing foam, cleansing water, pack, body oil, body lotion, oil-in-water makeup base, water-in-oil makeup base and foundation, but is not limited thereto.

When the composition of the present invention is a cosmetic composition, the cosmetic composition may include, in addition to the active ingredient, conventional adjuvants such as stabilizers, solubilizers, vitamins, pigments and flavors, and a carrier. In addition, the cosmetic composition may further include a skin absorption enhancer to enhance the effect.

The formulation of the cosmetic composition may be prepared in any formulation commonly prepared in the art, for example, the cosmetic composition may be a lotion, emulsion, skin, toner, lotion, essence, sunscreen, cream, makeup base, foundation , Powders, packs, gels, shampoos, rinses, sprays, makeup removers and cleansers, and the like, but are not limited thereto.

When the formulation is a paste, cream or gel, 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. .

If the formulation is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, especially in the case of a spray, additionally chlorofluorohydrocarbon, propane / butane Or propellants such as dimethyl ether.

If the formulation is a solution or emulsion, solvents, solubilizers or emulsions are used as carrier components, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3 Fatty acid esters of butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the formulation is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline cellulose as carrier components , Aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

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, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example 1 Preparation of Caffeine and Chlorogenic Acid

Coffee beans and roasting beans were used as ingredients. Specifically, the hot water extract is boiled in 100 ml of coffee 200 ml purified water to 100 ℃ and the temperature is lowered and extracted for 20 minutes at 80 to 90 ℃ the coffee is best extracted, centrifuged for 20 minutes at 15 ℃ 8,000 rpm After separation it was collected (FIG. 4). To separate caffeine from the extract, filter the residue from the hydrothermal extract and transfer the hydrothermal extract to the fraction funnel, add 200 ml of methylene chloride (Di-chlromethane) and 5 g of sodium chloride, mix and fractionate Methylene chloride layer was collected. Concentration under reduced pressure from the water layer was repeated to concentrate chlorogenic acid. To the water layer was added again 50 ml of methylene chloride per 100 g of green beans, and the fractions were collected repeatedly, and the methylene chloride layer was collected. The collected two methylene chloride layers were mixed, anhydrous sodium sulfate was added to remove residual moisture, filtered through a filter paper, and concentrated under reduced pressure. The concentrated crude extract was dissolved in an appropriate amount of 95% ethanol, cooled and recrystallized, filtered quickly with a Buchner funnel, and the recrystallized materials of the filter paper were recovered (FIG. 4). The recovered material was analyzed using a gas chromatography-mass spectrophotometer (GC-MS) (Fig. 2). As a result of the preparation of the extracts, it was confirmed that chlorogenic acid is concentrated 10-50%, caffeine was purified at least 95%.

All experiments were performed three times, the results of all the experiments were expressed as Mean ± Standard error, statistical analysis of experimental data was the significance of the test group relative to the control group by Student's t -test, when p -value is 0.05 statistical Judgment was significant.

Example 2: Confirmation of Antioxidant Activity of Extract

2-1. DPPH radical scavenging activity measurement

Antioxidant activity using DPPH radical scavenging activity was determined by modifying Blois' method (Blois ML. Antiocidant determination by the use of a stable free radical, Nature, 181, 1199 (1958)), which measured antioxidant activity. One of the methods that are used a lot. DPPH free reducing power was measured using DPPH (1,1-diphenyl-2-picrylhydrazyl) radical (SigmaAldrich CO., St. Louis, MO, USA) for the radical scavenging ability of the extract. Experimental method was mixed with 100 μL of 0.2 mM DPPH solution dissolved in ethanol in 200 μL of diluted sample at each concentration and reacted for 30 minutes at room temperature, and then 517 using ELISA reader (Infinite ™ F200, TECAN, Mannedorf, Switzerland). Absorbance was measured at nm. As a control, ethanol was treated instead of DPPH. In addition, ascorbic acid (AA) was used as a positive control. DPPH radical scavenging activity was calculated according to the following equation.

[Calculation 1]

As a result, as shown in Figure 5, it was confirmed that chlorogenic acid has a high radical scavenging ability in a concentration-dependent manner similar to ascorbic acid as a positive control. However, caffeine was found to have very low antioxidant activity.

Cell damage is caused by free radicals produced in vivo by oxidative stress, resulting in aging and disease. DPPH (2,2-diphenyl-1-picrylhydrazyl) is a free radical compound used to measure free radical scavenging activity and is used to measure antioxidant capacity using reducing compounds such as tocopherol and ascorbic acid, which are aromatic compounds and amines. DPPH, a stable model of free radicals, can be seen that the free radical scavenging reaction proceeds through the reduction of DPPH during the reaction. In addition, the degree of inhibition of the initial lipid peroxidation reaction can be predicted. Free radicals are also called harmful oxygen. Unsaturated fatty acids are constituents of cell membranes, and active oxygen accumulates lipid peroxidation by attacking unsaturated fatty acids to accumulate lipid peroxide in the body. This is known to cause deterioration of biological function and at the same time cause aging and adult diseases. In addition, antioxidant activity by various kinds of plant components and extracts has been reported.

In the present invention, the radical scavenging activity of caffeine, chlorogenic acid and coffee extract was measured using DPPH. As a result of antioxidant activity, chlorogenic acid and coffee extract showed free radical scavenging activity similar to the positive control, and caffeine showed concentration-dependent free radical scavenging activity. In the case of chlorogenic acid and coffee extract, DPPH radical scavenging ability was 97.22% and 89.99% in 50 μg / mL sample treatment, and 58.18% in caffeine in 1,000 μg / mL sample treatment (FIG. 5).

2-2. ABTS · ⁺  Scavenging activity measurement

ABTS [2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt] radical scavenging activity was measured by modifying the method of Re. ABTS (SigmaAldrich CO., St. Louis, MO, USA) was dissolved in distilled water at a concentration of 7 mM, and then mixed at a ratio of 1: 1 by adding 2.45 mM potassium persulfate to react in a dark room at room temperature for 24 hours. Used to form ABTS radicals. The radical-formed ABTS solution was diluted with ethanol and adjusted so that the absorbance measured at 734 nm was 0.70 ± 0.02. The scavenging ability was measured by mixing 150 μL of the ABTS solution and 150 μL of the sample for 6 minutes and then mixing the sample with each concentration at 734 nm using an ELISA reader (Infinite ™ F200, TECAN, Mannedorf, Switzerland). Ascorbic acid was used as a positive control. ABTS · ⁺ scavenging activity was calculated according to the following equation.

[Calculation 2]

In the present invention, the radical scavenging activity was measured using ABTS to investigate the more detailed antioxidant capacity of the extracts. A brief understanding of the measuring principle of the ABTS radical scavenging activity is that, first, the ABTS is oxidized by potassium persulfate to become cyan, and then the process of gradually decreasing the cyan as the electron donating ability is reduced by the antioxidant contained in the extract. How to measure. In addition, the ABTS cation scavenging method can measure both chain-cutting antioxidants and hydrogen donor antioxidants, and is applicable to both hydrophilicity and hydrophobicity. . In the present invention, using the ABTS was measured the radical scavenging activity of the hot water extract and ethanol extract of the coffee hide. ABTS ^{· +} radical scavenging ability of hot water and ethanol extract was 62.04%, 78.78%, 87.50%, 98.93, 99.61% according to the sample treatment of 50, 100, 250, 500, 1,000 ㎍ / mL Showed excellent antioxidant capacity of 92.26%, 99.64%, 99.09%, 98.09% and 100.34% at 50, 100, 250, 500, and 1,000 μg / mL. In addition, in the case of ethanol extract 50 ㎍ / mL it was confirmed that the results similar to ascorbic acid as a positive control (Fig. 6).

2-3. in vitro tyrosinase inhibitory activity

In vitro tyrosinase inhibition assay was performed to determine the degree of inhibition of the activity of tyrosinase (trosinase). In vitro tyrosinase inhibition assay was modified by Mason et al. (Mason HS, Peterson EW. Melanoproteins I. Reactions between enzyme-generated quinones and amino acids.Biochim. Biophys. Acta., 111, 134 (1965).) Measured. The test method was 250 μL of 0.1 M phosphate buffer (pH 6.5), 225 μL of the sample diluted at each concentration, 25 μL of mushroom tyrosinase (2000 U / mL) (SigmaAldrich CO., St. Louis, MO, USA) and 1.5 225 μL of mM tyrosine (SigmaAldrich CO., St. Louis, Mo., USA) was added sequentially, followed by reaction at 37 ° C. for 10 minutes. After the reaction was completed, the absorbance was measured at 490 nm using a UV / VIS Spectrophotometer (Optizen 2120UV, Mecasys, Daejeon, Korea). In this case, 0.1 M phosphate buffer solution (pH 6.5) was used instead of the sample solution. In addition, arbutin was used as a positive control. The tyrosinase activity inhibition rate was calculated according to the following formula.

[Calculation 3]

a: absorbance after reaction of blank sample liquid

b: absorbance after reaction of the sample liquid

a ', b': absorbance measured by replacing buffer

Melanin is a pigment produced in melanosomes, which are specialized organelles in melanocytes. Melanin is produced by the combined action of several enzymes such as tyrosinase, tyrosinase-related protein 1 (TRP1) and tyrosinase-related protein 2 (TRP2) in melanosomes. do. In particular, tyrosinase is a major factor acting in the process of hydroxylation of tyrosine, a rate-limiting step of melanin biosynthesis, to hydroxylation with dopa (3,4-dihydroxyphenylalanine, DOPA) and then to dopaquinone. It is a regulatory enzyme. Substances such as arbutin, hydroquinone, and vitamin C derivatives, which regulate and inhibit melanin synthesis, have been developed to prevent and resolve pigmentation, but do not show sufficient effects or are safe for the skin. Due to limitations such as stability in formulations, it has been prescribed in the manufacture of cosmetic formulations. In the tyrosinase inhibitory effect of each sample, caffeine showed 29.24% and 50.47% 66.82% in 100, 1,000 and 10,000 ㎍ / mL sample treatment, and chlorogenic acid in 100, 1,000 and 10,000 ㎍ / mL sample treatment, respectively. The results were 25.04%, 38.35%, and 49.31%. The coffee extract showed 20.28%, 23.46%, 28.11% tyrosinase inhibitory effect at 100, 1,000, 10,000 μg / mL sample treatment (FIG. 7).

2-4. Cytotoxicity and Proliferation Test

Human keratinocyte (HaCaT) (ATCC, Manassas, VA, USA) and mouse melanocytes (B16F10) (Korea Cell Line Bank, Seoul, Korea) are Dulbecco's Modified Eagle Medium (DMEM) (HyClone, Logan, UT, USA) medium, human fibroblast (Detroit551) (Korea Cell Line Bank, Seoul, Korea) used Minimum Essential Medium (MEM) (WelGene, Gyeongbuk, Korea) medium, 10% fetal bovine serum (fetal) bovine serum (FBS) (HyClone, Logan, UT, USA) and penicillin (100 units / mL) / streptomycin (100 μg / mL) (HyClone, Logan, UT, USA) were added to the CO ₂ cell incubator (NU- 4750G, NuAire, Plymouth, MN, USA) was incubated under 5% CO ₂ supply, and passaged at 2-3 days intervals.

Each sample diluted by concentration was evenly divided into 96 well plates at 5 x 10 ³ cells / well, and after 24 hours, each extract was incubated for 72 hours. Effect on proliferation of cells by the treatment concentrations were measured using the ^{Cell Titer 96 ⓡ AQueous One Solution Cell} proliferation assay kit (. Promega, Madison, Wis, USA). After incubation, all medium was removed and 100 μL of DMEM medium and 3- (4,5-dime thylthiazol-2-yl) -5- (3-carboxymethoxymethoxy) -2- (4-sulfophenyl) -2H-tetrazolium inner salt 20 μL of (MTS) solution was added thereto, followed by reaction for 3 hours in a dark condition at 5% CO ₂ , 37 ° C., and then absorbance was measured at 490 nm using an ELISA reader (Infinite ™ F200, TECAN, Mannedorf, Switzerland). Untreated culture was used as a control. Cell viability was calculated according to the following equation.

[Calculation 4]

As a result of measuring the cell proliferation rate by treating the concentration of caffeine, chlorogenic acid, and coffee extract by concentration, the cell proliferation rate was as shown in FIG. 95% purified caffeine reduced cell viability at high concentrations (1 mg / ml) (Fig. 8). However, chlorogenic acid and coffee extract showed the best results at 100 μg / mL and cell survival rates of 100.5% and 110.7%, respectively (FIG. 8).

2-5. Hair Loss Prevention Evaluation

Dihydrotestosterone (DHT), a prostatic androgen that is a male hormone in the prostate, is known to cause prostate hypertrophy as well as hair loss in men (Aggarwal et la., 2009). Prostate DHT is a metabolite of testosterone that is converted by 5-a reductase (Shin et al., 2012). DHT binds to androgen receptors in cells, resulting in an androgen effect (Chaudhary and Turner 2010). In particular, since DHT has a higher affinity for binding to androgen receptors (ARs) than testosterone, it acts as a strong active hormone, increasing the size of the prostate gland. In prostate hyperplasia, DHT levels in the blood are measured high (Wang et al., 2010).

The enzyme that converts testosterone to DHT is 5-α reductase. Humans have type 1 5-α reductase (SRD1, SRD5A1, type I 5-α SRD) and type 2 5-α reductase (SRD2, SRD5A2). , type II 5-α SRD). Type 1 5-α reductase is mainly distributed in the liver, sebaceous glands, non-genital skin, prostate, scalp and brain. Type 2 5-α reductase is mainly distributed in prostate tissue, genital skin, ejaculatory canal, liver, hair root, uterus, breast and brain (Pais 2010). In particular, the activity of 5-α reductase converts testosterone to DHT, which contributes to normal male growth. However, excessively high activity of 5-α reductase increases the content of DHT, which induces diseases such as acne, histism, irritable alopecia, benign prostatic hyperplasia, and enlarged prostate gland (Figure 3) (Wilson, 1968). . To overcome this, chlorogenic acid and caffeine were tested for hair loss prevention and hair growth.

Example 3: Animal Experiment

end. Laboratory animals

Seven-week-old male C57BL / 6 mice were distributed at Yeungnam University Experimental Animal Center and were regularly fed with food and water under a 12-hour photoperiod cycle. This experiment was approved by Yeungnam University Research Ethics Committee.

I. Substance prescription and hair growth test

The method of hair growth determination was confirmed to the extent of hair growth that promotes the activity of the extract (Roh et al. 2002; Kumar et al 2012). Twenty four 7-week-old C57BL / 6 mice were randomly divided into four isolates for each test group and divided into four groups, one group for each positive control and one vehicle control. The back of each test mouse was dehaired 2 cm x 3 cm wide using a hair clipper. In order to remove hair follicles and fine hairs remaining in the skin, hair removal cream (Nicklin Cream, Ildong Pharmaceutical) was evenly applied and after 5 minutes, hair removal was completely removed using a hair removal spatula. It was reared 24 hours for recovery. The delivery solution (vehicle) was prepared with 1,3 Butylen Glycol: water: ethyl alcohol (95%) in a ratio of 5:45:50. The test solution in the delivery solution consisted of 0.1% minoxidil (control), coffee bean extract, refined caffeine, and 50% chlorogenic acid containing coffee beans (CGA50) 0.1%, purified caffeine 95% + chlorogenic acid 0.1% for each test group. It was contained in the transfer solution. For the mixed prescription test group, 0.1% of chlorogenic acid and 0.1% of GBE-CGA50 were contained, respectively. Only untreated control was used as the transfer solution. 100 μl of the test solution was applied to the hair removal site of each test group for 3 weeks at a constant time every day.

As a result, the hair growth evaluation confirmed that the growth of the hair was promoted by the treated material, and the leather was black and grayed. Hair growth in the anagen phase of hair follicles was measured. The degree of hair growth per depilation area is expressed as a percentage. Hair growth scores were assigned from 1 to 5 points. In short, 0 = no observed growth; 1 = up to 20% growth; 2 = 20-40% growth; 3 = 40-60% growth; 4 = 60-80% growth; 5 = 80% -100% growth was set.

As observed in Figure 9, caffeine and chlorogenic acid showed a hair growth effect. In addition, as a result of treating the complex material of caffeine and chlorogenic acid showed a very high effect of hair growth similar to minoxidil (Fig. 9).

Therefore, in the present invention, it was confirmed that caffeine and chlorogenic acid exhibit excellent hair growth effects as desired.

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

The scope of the present invention is represented by the following claims, and it should be construed that all changes or modifications derived from the meaning and scope of the claims and their equivalents are included in the scope of the present invention.

Claims (3)

Cosmetic composition for improving scalp health containing natural caffeine and chlorogenic acid as an active ingredient. The cosmetic composition for improving scalp health of claim 1, wherein the natural caffeine and chlorogenic acid are included in an amount of 0.0001 to 50 parts by weight based on 100 parts by weight of the total composition. The cosmetic composition according to claim 1, wherein the cosmetic composition is a shampoo, hair treatment, hair tonic, flexible lotion, gel, water soluble liquid, milk lotion, cream, essence, skin toner, oil-in-water type. Formulated in a formulation selected from the group consisting of in-water emulsions, water-in-oil emulsions, cleansing foams, cleansing water, packs, body oils, body lotions, oil-in-water makeup bases, water-in-oil makeup bases and foundations Cosmetic composition for improving scalp health, characterized in that it is.

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