KR101721666B1 - Skin-whitening compound from Inula britannica, and skin-whitening composition containing the same - Google Patents

Abstract

The present invention relates to a skin-whitening compound and a composition for skin-whitening containing the same, and more particularly, the present invention provides innoramo that is a sesquiterpene lactone-type compound, which provides an excellent melanin synthesis inhibitory effect and an anti-inflammatory effect, at the same time, and does not irritate skin, and a novel skin-whitening composition containing the same as an active ingredient.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a skin whitening compound and a skin whitening composition containing the same,

The present invention relates to a skin whitening compound and a composition for skin whitening comprising the same, and more particularly, to a skin whitening compound having an excellent melanin synthesis inhibitory effect and an anti-inflammatory effect, (innoramo) and a novel skin whitening composition containing the same as an active ingredient.

Human skin is the largest organ in the human body, and it plays important physiological functions such as secretion, respiration, and synthesis as well as physical and chemical protection. Such skin color is largely determined by melanin, hemoglobin, carotene and the like. Among them, melanin is the most important factor. In particular, melanin is produced in melanosomes in the melanocytes in the basal layer of the mammalian skin and is synthesized as a defensive means to protect the living body and has a decisive influence on the color of the skin color, hair, eyes.

Melanin produced by the skin partially disappears through skin physiology metabolism. However, natural aging, photoaging, and immune degradation cause a large amount of melanin on the skin to remain on the skin, thereby promoting skin aging or inducing skin cancer. The resulting melanin leads to pigmentation such as spots, freckles, and black spots, which not only becomes a cosmetic problem but also a fatal cause of skin aging.

The synthesis of melanin is influenced by ultraviolet light, cytokines, growth factors and hormones, and they are produced through various complex signal transduction in the cells. The main pathway is cAMP (cyclic adenosine monophosphate) pathway and PKA A) The path is well known. In other words, when the skin receives ultraviolet rays, it increases the cAMP signal of melanocytes, then affects the signal transduction substance PKA and affects the intracellular cAMP response element binding (CREB) protein, and secretes the microphthalmia-associated transcription factor It affects regulation. Therefore, the melanin synthesis inhibitor is widely used as a skin treatment agent in the pharmaceutical industry and as a skin whitening agent for preventing and treating spots and freckles in the cosmetics industry.

Tyrosinase, tyrosinase related protein-1 (TRP-1) and tyrosinase related protein-2 (TRP-2) are known enzymes involved in intracellular melanin synthesis. These melanin-related enzymes greatly affect the color of the face, which can be broadly divided into the yellow-based phomelanin and the black-based eumelanine.

Melanin is synthesized by firstly converting tyrosine into DOPA (3,4-dihydroxyphentl-alanine) by tyrosinase and then oxidizing it to DOPA quinone and converting DHI (5,6-di Hydroxyindole) and indole-5,6-quinone are oxidized to produce melanin. The resulting melanin is transferred to the keratinocyte through the melanosome vesicle and then excreted through the keratinization process to the skin surface, along with the keratin. However, in this process, melanin is produced in excess and melanin is not completely eliminated by keratinization, pigmentation phenomenon occurs. Therefore, in order to prevent such pigmentation phenomenon, it is necessary to control a part of melanin production to reduce the production of melanin.

There are many mechanisms that inhibit the synthesis of melanin. First, a typical method is to inhibit the activity of tyrosinase, which is an important enzyme of melanin synthesis. The most known one is a chelator such as kojic acid which chelates copper ions necessary for enzyme activity and inhibits tyrosinase activity have. As plant extracts inhibiting tyrosinase activity, extracts of Eucalyptus species and licorice extracts are known. On the other hand, there is a method of reducing melanin synthesis using a substrate-like substance which is a competitive inhibitor of tyrosine, which is a substrate of tyrosinase, and arbutin and the like are known as such substrate-like substances.

However, these whitening raw materials have a disadvantage in that their use is limited because their efficacy is relatively weak, their stability is low, their effects are not sustained for a long time, and they are highly irritating to skin. Therefore, it is urgent to find new whitening materials.

On the other hand, fleet Chemistry (旋覆花) is in geumbulcho (Inula Linne), a kind of medicinal plants geumbulcho (Inula britanica Linne belonging to the ssp . japonica Kitamura )) is called as a lavender flower. It is a perennial plant of 20 ~ 60 ㎝ in height. It grows in rhizomes and grows yellow with a diameter of about 3 ㎝. In addition to the genus Chrysomelidae, the genus Chrysanthemum ( Inula britanica Linne var. Linariaefolia Regel and Inula britanica I have linne . ramosa Kom ) are known.

Inula britanic ) is a golden flower in the shape of chrysanthemum. It is also called "草" in Korean medicine. It is also called "夏 菊" because it blooms in summer. It is well known that it is effective for coughing, shortness of breath, or treatment of chronic gastritis.

To date, the efficacy of extracts or fractions of L. monocytogenes has been limited to compositions for the treatment of gastric ulcers (Korean Patent Laid-open Publication No. 10-2016-0089618), compositions for the treatment of highly pathogenic avian influenza (Korean Patent No. 10-1195120), compositions for the treatment of pancreatic cancer Korean Patent Publication No. 10-1503585), a cosmetic composition for alleviating scalp irritation (Korean Patent Laid-Open Publication No. 10-201507), a composition for treating inflammation and allergy (Korea Patent No. 10-1150643) 10-1397169), antioxidant activity and cosmetic composition (Korean Patent No. 10-1405128), brain function and cognitive function improving effect (Korean Patent No. 10-0569089), angiogenesis inhibitory effect (Korean Patent No. 10-0500298) . In addition, whitening efficacy of 1-O-acetyl-4R, 6S-bithanyllactone and 1-hydroxy-4R, 6S-bithanyllactone, which are lactone compounds, has been reported (Korean Patent No. 10-1050484).

The present inventors have while trying to navigate the whitening material from domestic natural resources, the kind geumbulcho (Inula britannica var. Ramosa Kom ) for the first time confirmed that there is a very good whitening effect from a compound that has no known whitening effect (hereinafter referred to as "innoramo"), and has completed a series of experiments, The whitening effect was confirmed, and it was confirmed that it is a very safe compound which does not show toxicity on the skin, and the present invention has been completed.

Korean Patent Registration No. 10-1503585 (Feb. Korean Patent Registration No. 10-1050484 (July 13, 2011)

It is an object of the present invention to provide Inoram, which is a skin whitening compound which has excellent melanin synthesis inhibitory effect and anti-inflammatory effect, but which is not skin irritant.

It is still another object of the present invention to provide a skin whitening cosmetic composition comprising Inoramo, which is a skin whitening compound, and a pharmaceutical composition for skin whitening.

It is another object of the present invention to provide a method for extracting an effective ingredient for skin whitening and an extract for skin whitening extracted therefrom in order to obtain Inolamo, a skin whitening compound.

In order to accomplish the above object, one aspect of the present invention relates to a skin whitening compound represented by the following general formula (1).

[Chemical Formula 1]

Further, in an embodiment according to the present invention, the skin whitening compound may be one isolated from the embryo.

In an embodiment of the present invention, the skin whitening compound may be one for inhibiting melanin synthesis.

Another aspect of the present invention relates to a skin whitening cosmetic composition comprising the skin whitening compound.

Another aspect of the present invention relates to a pharmaceutical composition for skin whitening comprising the skin whitening compound.

Another aspect of the present invention is a method for producing an ethanol extract, comprising the steps of: a) extracting an egg yolk with ethanol to obtain an ethanol extract; And b) sequentially adding hexane and chloroform to the ethanol extract and fractionating to obtain a chloroform fraction containing a compound represented by the following formula (1): (1) extracting an effective ingredient for skin whitening; Whitening extract.

[Chemical Formula 1]

In another embodiment of the present invention, the step a) may be carried out at 30 to 100 ° C for 5 to 24 hours.

In another embodiment of the present invention, the method may further comprise the step of: c) purifying the chloroform fraction through column chromatography to obtain the compound represented by Formula 1.

Inoramo, which is a sesquiterpene lactone-based compound according to the present invention, has excellent melanin synthesis inhibitory effect and anti-inflammatory effect, but has no skin irritation and is used as a skin whitening cosmetic composition or a skin whitening pharmaceutical composition There is an advantage that it is possible.

FIG. 1 is a view showing the chemical structure of Inolamo which is a skin whitening compound according to the present invention.

Hereinafter, the skin whitening compound of the present invention and the skin whitening composition containing the same will be described in detail with reference to the accompanying drawings. The following drawings are provided by way of example so that those skilled in the art can fully understand the spirit of the present invention. Therefore, the present invention is not limited to the following drawings, but may be embodied in other forms, and the following drawings may be exaggerated in order to clarify the spirit of the present invention. Also, throughout the specification, like reference numerals designate like elements.

Hereinafter, the technical and scientific terms used herein will be understood by those skilled in the art without departing from the scope of the present invention. Descriptions of known functions and configurations that may be unnecessarily blurred are omitted.

The present invention relates to a sesquiterpene lactone-based compound having both excellent melanin synthesis inhibitory effect and anti-inflammatory effect and having no skin irritation. More specifically, the present invention relates to a skin whitening compound represented by the following general formula (1).

[Chemical Formula 1]

The skin whitening compound represented by Formula 1 suppresses the action of tyrosinase, an enzyme that plays an important role in melanin synthesis in cells, and effectively inhibits the synthesis of melanin, thereby exhibiting an excellent skin whitening effect. That is, the skin whitening compound represented by the formula (1) can be used for inhibiting melanin synthesis.

The skin-whitening compound represented by the formula (1) may be one isolated from the fortification. For example, the extract obtained from the fortification may be fractionated and purified to provide a skin-whitening compound of high purity, And an anti-inflammatory effect at the same time, it may not stimulate the skin.

With such an effect, a skin whitening cosmetic composition or a skin whitening pharmaceutical composition can be provided using the skin whitening compound represented by the formula (1).

First, the present invention can provide a cosmetic composition for skin whitening comprising the compound for skin whitening represented by the general formula (1).

In one example of the present invention, the amount of the skin whitening compound represented by the formula (1) is not particularly limited as long as the effect of inhibiting melanin synthesis can be exhibited without compromising the physical properties of the cosmetic composition to be provided, , The amount of the skin whitening compound represented by the formula (1) may be 0.01 to 90% by weight, more preferably 1 to 75% by weight, and more preferably 5 to 60% by weight based on the total weight of the composition However, the present invention is not limited thereto.

Cosmetics made from cosmetic compositions for skin whitening may be those which comprise both conventional emulsified formulations or solubilized formulations. Cosmetics of emulsified form include nutritional lotion, cream or essence, and cosmetics of solubilized form have softening longevity. It can also be prepared in the form of adjuvants which can be applied topically or systemically, which are conventionally used in the field of dermatology by containing a dermatologically acceptable medium or base.

As the formulations of suitable cosmetics, the solubilized formulations may be solutions, gels, solid or paste anhydrous products, and the transfer formulations may include aqueous emulsions, suspensions, microemulsions, microcapsules, microgranules, ionic Or a non-ionic follicular dispersing agent (liposome). The final cosmetic formulations may be presented in the form of creams, skins, lotions, powders, ointments, sprays or conceal sticks. It can also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

In addition, the cosmetic composition for skin whitening according to an exemplary embodiment of the present invention may further comprise at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents, , Water, ionic or nonionic emulsifiers, fillers, sequestering agents and chelating agents, preservatives, vitamins, barrier agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics And may further contain adjuvants commonly used in the cosmetics or dermatology fields, such as any other ingredients used. And the above ingredients may be introduced in amounts commonly used in the dermatology field.

In addition, specific formulations of the skin whitening cosmetic composition according to an exemplary embodiment of the present invention may include a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, It includes formulations such as creams, essences, nutritional essences, packs, soaps, shampoos, cleansing foams, cleansing lotions, cleansing creams, body lotions, body cleansers, emulsions, press powders, loose powders, eye shadows and the like.

In addition, the present invention can provide a pharmaceutical composition for skin whitening comprising the skin whitening compound represented by the general formula (1). Such a pharmaceutical composition for skin whitening can have a skin whitening performance as well as an inflammation treatment effect.

In one embodiment of the present invention, the addition amount of the skin whitening compound represented by the general formula (1) is not particularly limited as far as the effect of inhibiting melanin synthesis or the effect of treating inflammation can be expressed. However, The amount of the whitening compound to be added may be 0.01 to 90% by weight, more preferably 1 to 75% by weight, and still more preferably 5 to 60% by weight based on the total weight of the composition, but is not limited thereto .

The pharmaceutical composition for skin whitening may contain at least one active ingredient which exhibits the same or similar functions in addition to the skin whitening compound represented by the general formula (1).

The pharmaceutical composition for skin whitening according to an exemplary embodiment of the present invention may further include pharmaceutically acceptable additives. Examples of the pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, lactose, Wherein the active ingredient is selected from the group consisting of povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, mannitol, glutinous rice gum, arabic gum, pregelatinized starch, corn starch, powdered cellulose, hydroxypropylcellulose, One or more selected from aluminum, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, white sugar, dextrose, sorbitol and talc may be used. The pharmaceutically acceptable additives according to the present invention are preferably included in the composition in an amount of 0.1 to 90% by weight, but are not limited thereto.

On the other hand, as mentioned above, inoramo, which is a skin whitening compound represented by the general formula (1), may be one isolated from lyophilization and may be one obtained by fractionating and purifying the extract obtained from lyophilization , A method for extracting an effective ingredient for skin whitening represented by the formula (1) from the embryo is described in more detail.

A method for extracting an effective ingredient for skin whitening according to an embodiment of the present invention comprises the steps of: a) obtaining an ethanol extract by extracting with an ethanol; And b) sequentially adding hexane, chloroform and ethyl acetate to the ethanol extract and fractionating to obtain a chloroform fraction containing a compound represented by the following formula (1).

[Chemical Formula 1]

Through the above extraction method, innoramo, which is a sesquiterpene lactone-based compound having excellent melanin synthesis inhibitory effect and anti-inflammatory effect and having no skin irritation, can be obtained.

In one example of the present invention, the embedding of step a) may be carried out in the wild or commercially available, but is not limited thereto.

The ethanol in step a) is not particularly limited as long as it contains ethanol, but it is preferable that the purity is 50 to 100% by weight, more preferably 75 to 100% by weight, more preferably 90 to 100% by weight, particularly preferably 98 to 100% Use of ethanol is preferable for more effective extraction of Inoramo.

The amount of ethanol to be added is not particularly limited, but is preferably 1 to 10 times the volume of the supernatant to be extracted, more preferably 2 to 5 times the extract.

In addition, in one embodiment of the present invention, the step a) is preferably performed at 30 to 100 ° C for 5 to 24 hours, more preferably at 40 to 60 ° C for 10 to 20 hours. In this range, Can be extracted.

The number of times of extraction is preferably 1 to 5 times, and more preferably 2 to 3 times, but not always limited thereto. Any of the conventional extraction methods such as hot water extraction, hot water extraction, ultrasonic extraction, and reflux extraction may be used as the extraction method.

The step b) is a step of first fractionating the extract extracted through a) according to the physical properties of the compound. First, the ethanol extract is first extracted using hexane to remove nonpolar impurities having extremely low polarity . The number of times of fractionation using hexane is not particularly limited, but it is preferably 1 to 5 times, more preferably 2 to 3 times.

Next, the residue is extracted with hexane using chloroform, and the remaining filtrate is further fractionated. From this, fractions containing the inoramo that are to be obtained in the present invention can be obtained. This is also not particularly limited, but it is preferable to repeatedly carry out fractionation 1 to 5 times, more preferably 2 to 3 times, so that the Inolamo can be sufficiently fractionated.

Thereafter, the filtrate is further extracted with chloroform using ethyl acetate, and the remaining filtrate is further fractionated. Then, it can be confirmed whether or not the inner layer remains in the filtrate. If the inner layer remains, the inner layer can be separated from the filtrate It is of course possible to further carry out the step of

In addition, column chromatography may be further performed to obtain an enolamo having a higher purity after step b). More specifically, the method for extracting embryos according to an example of the present invention may further comprise the step of: c) purifying the chloroform fraction through column chromatography to obtain the compound represented by the above formula (1).

In step c) according to an example of the present invention, column chromatography can be carried out through silica gel, Sephadex, or RP-18 or the like to separate and purify Inolamo. The column chromatography can be carried out several times by selecting an appropriate filler as necessary. The solvent may be hexane-ethyl acetate, chloroform-ethanol, ethyl acetate-ethanol or dichloromethane-ethanol.

In addition, further reverse phase column chromatography, Sephadex LH-20 column chromatography or preparative HPLC may be performed to obtain higher purity Inolamo, but the present invention is not limited thereto.

In addition, the present invention can provide an extract for skin whitening extracted from the method for extracting an active ingredient for skin whitening as described above. Specifically, the present invention provides a skin whitening extract comprising the steps of: a) obtaining an ethanol extract by extracting with ethanol; ; And b) sequentially adding hexane, chloroform and ethyl acetate to the ethanol extract and fractionating to obtain a chloroform fraction containing a compound represented by the following formula 1: An extract for skin whitening can be provided.

Hereinafter, the skin whitening compound and the skin whitening composition containing the same according to the present invention will be described in more detail with reference to the following examples. It should be understood, however, that the invention is not limited to the disclosed embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the invention. Unless otherwise defined, all technical and scientific terms have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Also, the singular forms as used in the specification and the appended claims are intended to include the plural forms as well, unless the context clearly indicates otherwise. In addition, the unit of the additives not specifically described in the specification may be% by weight.

[Example 1]

5 kg of Inula britannica was purchased from Gwangdeok Pharmaceutical located in Chubu-myeon, Jeonbuk-gun, Chungnam Province and 50 L of 100% purity ethanol was added. Using MS-DM609 / 20L and KSP-248L, Time reflux extraction. The obtained extract was filtered and then ethanol was evaporated using a vacuum concentrator (EYELA N-12) to prepare 275 g of a lyophilized ethanol extract.

Next, 275 g of the supernatant ethanol extract was suspended in 8 L of water, added with hexane (8 L), extracted three times using a separatory filter, and then the remaining aqueous layer was extracted with chloroform (8 L) three times. To the remaining aqueous layer was added water (2 L) and extracted three times with ethyl acetate (8 L). By concentrating the hexane layer, the chloroform layer and the ethyl acetate layer, respectively, 90 g of hexane fraction, 120 g of chloroform fraction and 50 g of ethyl acetate fraction were obtained.

Next, a step gradient solvent system consisting of 120 g of the chloroform fraction and silica gel column chromatography with hexane-ethyl acetate (100: 0 to 0: 100) was applied to obtain five fractions. After fractionating the fractions exhibiting melanin synthesis inhibitory activity from the fractions obtained above, the fractions were subjected to Sephadex LH-20 column chromatography to obtain active fractions.

Next, the active fractions were subjected to Sepatex LH-20 column chromatography with different solvent systems, and the resulting product was recrystallized with acetone to obtain 280 mg of pure crystalline phase inulin. The thus-obtained compound was confirmed to be a sesquiterpenactone compound (Inoramo) represented by the general formula (1) through instrumental analysis such as NMR.

Appearance: white powder

Melting point: 156.3 ~ 157.7 ℃

Molecular weight: 536.32 g / mol

HR-FAB-MS: 537.382 (M + H) ⁺ (calculated: C ₃₂ H ₄₁ O ₇ : 537.396)

¹ H NMR (CDCl ₃ , 400 MHz):? Ppm 3.80 (1H, m, H-1a), 3.58 m, H-2b), 1.50 (1H, m, H-3a), 1.40 (1H, m, H-3b), 2.53 , 7.00 (1H, d, J = 8.0 Hz, H-8), 7.07 (1H, d, J = 8.0 Hz, H- d, J = 11.9 Hz, H-13b), 2.34 (3H, s, H-14), 0.98 M, H-6'b), 2.66 (1H, m, H-6 '), 2.77 H-7 '), 4.11 (1H, ddd, J = 3.0,8.8,11.5 Hz, H-8'), 2.29 (1H, dt, J = 4.3, 12.1 Hz, H- (1H, m, H-9'b), 2.07 (1H, m, H-10 '), 6.12 (1H, d, J = 3.4Hz, H-13'a), 5,46 H-13 '), 1.01 (3H, d, J = 7.1 Hz, H-14'), 1.59 (3H, s, H- .

¹³ C NMR (CDCl ₃ , 500 MHz):? Ppm 62.35 (C-1), 30.47 (C-2), 32.98 (C-3), 34.29 C-6), 138.95 (C-7), 123.14 (C-8), 129.16 (C-14), 18.27 (C-14), 21.97 (C-15), 60.25 (C-5 '), 25.47 (C-6'), 45.28 (C-7 '), 82.19 (C-12 '), 118.87 (C-13'), 16.15 (C-14 '), 14.20 (C-15'), 171.00 C-2 ").

[Comparative Example 1]

5 kg of Inula britannica was purchased from Gwangdeok Pharmaceutical located in Chubu-myeon, Jeonbuk-gun, Chungnam Province and 50 L of 100% purity ethanol was added. Using MS-DM609 / 20L and KSP-248L, Time reflux extraction. The obtained extract was filtered and then ethanol was evaporated using a vacuum concentrator (EYELA N-12) to prepare 275 g of a lyophilized ethanol extract.

[Comparative Example 2]

1.5 kg of Inula britannica was purchased from Guangdong Pharmaceutical, located in Chubu-myeon, Jungsan-gun, Chungnam Province, and then immersed in 3 L of 100% pure methanol for 48 hours and repeatedly extracted three times using a filter. The obtained extract was evaporated to methanol using a vacuum concentrator to prepare 70 g of a supernatant methanol extract.

Next, the supernatant methanol extract (70 g) was suspended in 2 L of water, and hexane (2 L) was added thereto. The mixture was extracted three times using a separatory funnel, and then the remaining aqueous layer was extracted three times with chloroform (2 L) . Water (500 ml) was added to the remaining aqueous layer and extracted three times with ethyl acetate (2 L). The hexane fractions (25 g), the chloroform fractions (18 g) and the ethyl acetate fractions (10 g) were obtained by concentrating the hexane layer, the chloroform layer and the ethyl acetate layer, respectively.

The fraction obtained by concentrating the chloroform layer (18 g) was subjected to silica gel column chromatography (silica gel column, 20 g) in a stepwise gradient solvent system consisting of hexane-ethyl acetate (100: 0 to 0: Fr.1 to Fr.51) fractions were obtained. Of the fractions obtained above, Fr.51 showed melanin synthesis inhibitory activity and was screened.

This was subjected to Sephadex LH-20 column chromatography to obtain an active fraction.

Next, the active fractions were subjected to Sepatex LH-20 column chromatography with different solvent systems, and the resulting product was recrystallized with acetone to obtain 70 mg of a pure crystalline compound. The compound thus obtained was confirmed to be 1-hydroxy-4R, 6S-bithanyllactone (IB3, 1-hydroxy-4R, 6S-britannilactone) through NMR analysis and the like. A more specific extraction method of IB3 can be found in Korean Patent No. 10-1050484.

[Comparative Example 3]

Arbutin, known as a whitening agent, was prepared.

[Experimental Example 1] Analysis of melanin synthesis inhibitory activity of inoramo compound

<1-1> Culture of B16 melanoma cells

As a cell line producing melanin, B16F10 melanoma cells, which are generally used for screening whitening substances, were cultured in the Korean Cell Line Bank. The melanoma cell line B16F10 was mixed with DMEM (Dulbecco's Modified Eagle's Medium) medium supplemented with 10% fetal bovine serum (FBS), 100 μg / ml streptomycin and 100 U / ml penicillin, At a density of 1 × 10 ⁵ cells / well, and cultured at 5 vol% CO ₂ and 37 ° C for one day. Melanoma cells were cultured and passaged when 100-mm Patri dishes were about 90% by volume full.

<1-2> Measurement of melanin inhibition

The melanin synthesis inhibitory effect of Example 1 and Comparative Examples 1 to 3 was measured using cultured B16F10 melanoma cells. The experimental group treated with dimethylsulfoxide (DMSO) alone, the control group treated with 1, 5, 10, 50, and 100 ppm, respectively, and the control group of Comparative Example 3 were treated with 1, 5, 10, 50, and 100 ppm, respectively. Namely, cells were inoculated on a 24-well plate at a density of 1 × 10 ⁵ cells / well and cultured for one day. Then, the cells were exchanged with a fresh medium containing 100 nM of α-MSH (mealanocyte stimulating hormone), and the samples were treated at 1, 5, 10, 50 and 100 ppm for 72 hours. After 72 hours, the amount of melanin present in the medium was measured at 400 nm, and the melanin inhibitory activity of each fraction was measured to determine the IC ₅₀ value.

As a result, Inoramo inhibited the synthesis of melanin in a concentration-dependent manner by decreasing the amount of melanin synthesis with increasing concentration in the experiment for inhibiting melanin synthesis using B16 melanoma cells. In particular, as shown in the following Table 1, inolamo of Example 1 exhibited remarkable melanin inhibitory activity as compared with Comparative Examples 1 to 3. Thus, it can be seen that Inoramo, a sesquiterpene lactone compound according to the present invention, has a very excellent melanin synthesis inhibitory activity.

sample Melanin synthesis (%) Melanin synthesis inhibition IC ₅₀ (ppm) Negative control group
(Sample and group not treated with? -MSH) 22.45 ± 0.77 - Positive control group
(Group treated with &lt; RTI ID = 0.0 &gt; a-MSH &lt; / RTI & 100.00 3.44 -
Example 1 Inoramo
(ppm) One 100.51 + - 0.84 13.2 5 83.11 + - 0.53 10 58.75 ± 1.12 50 23.50 ± 1.42 100 8.59 ± 0.96 Comparative Example 1 Ethanol extract
(ppm) One 98.82 ± 1.43 55.4 5 97.90 + - 9.53 10 72.26 + - 8.74 50 59.01 + - 2.10 100 45.23 + - 3.43 Comparative Example 2 IB3
(ppm) One 101.26 + - 0.41 22.6 5 82.23 + - 1.22 10 75.90 ± 0.96 50 40.56 占 1.21 100 17.67 ± 0.98 Comparative Example 3 Arbutin
(ppm) 500 54.01 + - 3.05 805.7

[ Experimental Example  2] Cytotoxicity analysis

The compound of Example 1 and Comparative Example 1 To determine if it could be used as a whitening composition, cytotoxicity was investigated. First, the B16F10 melanoma cell line was mixed with DMEM (Dulbecco's Modified Eagle's Medium) medium supplemented with 10% fetal bovine serum (FBS), 100 μg / ml streptomycin and 100 U / ml penicillin, to 1 × 10 ⁵ it was inoculated at a density of cell / well. Then, Example 1 and Comparative Example 1 were added at concentrations of 1, 5, 25, 50, 100 and 250 ppm, respectively, and cultured for 24 hours. As a negative control group, a group not treated with the sample was used. After 24 hours, 10 μl of 5 mg / ml MTT (3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2H-tetrazolium bromide) Lt; / RTI &gt; Thereafter, after the medium and the MTT reagent were removed, 1 ml of DMSO was added to measure the degree of color development at 575 nm. This was repeated three times and then statistically processed to obtain an average value.

As a result, Inolamo showed high cell survival rate even at high concentration in the cytotoxicity measurement experiment using melanoma cell line B16, indicating that it is a very safe compound since it does not show cytotoxicity. In detail, as shown in Table 2, Example 1 and Comparative Example 1 showed almost no cytotoxicity even at a concentration of 100 ppm. Therefore, it can be seen that the inoramo compound of the present invention does not cause toxicity to the cells and can be safely used for human body.

sample Concentration (ppm) Cell survival rate (%) Control group (untreated group) 0 100
Example 1 Ethanol extract One  99.62 ± 1.78 5 101.93 + - 2.55 10 103.54 + - 2.13 50 102.29 + - 1.04 100 103.83 + - 2.40 Comparative Example 1 Inoramo One 100.98 + - 1.85 5 101.39 + - 2.25 10 102.48 + - 2.37 50 100.33 + - 1.91 100 103.58 ± 1.67

[ Experimental Example  3] Anti-inflammatory performance analysis

In order to confirm the anti-inflammatory activity of Inoramo obtained in Example 1, the TNF-alpha production inhibitory activity and the nitric oxide production inhibitory activity, which are tumor necrosis factors as inflammatory reaction factors, were analyzed.

<3-1> Inhibitory activity of TNF-alpha production

RAW264.7 cells were mixed in RPMI 1640 medium supplemented with 100 U / ml of penicillin, 100 μg / ml of streptomycin, 10% of petal bovine serum (rabbit serum) and cultured in 96-well plates at 1 × 10 ⁴ cells / Respectively. RAW264.7 cells were then treated with 100 ppm of Inolam and incubated for 18 hours in the presence of LPS (1 [mu] g / [mu] l). The amount of TNF-alpha produced was recovered from the supernatant and measured with OptELA ^th assay kit, the concentration of which was assessed by IC ₅₀ . As a negative control group, a group not treated with the sample was used.

As a result, the IC ₅₀ was measured to be about 54.39 μM.

<3-2> Nitric oxide production inhibitory activity assay

RAW264.7 cells were mixed in RPMI1640 medium supplemented with 100 U / ml of penicillin, 100 μg / ml of streptomycin and 10% of petal bovine serum (rabbit serum), and the cells were plated in a 96-well plate at 1 × 10 ⁵ cells / Respectively. After 3 hours, RAW 264.7 cells were treated with 100 ppm of Inolam in the presence of LPS (1 / / ㎕) or in the absence of LPS, followed by incubation for 24 hours. The culture medium was centrifuged, and 100 μl of a grease reagent (Griess reagent: 1% by weight of sulfanilamide, 0.1% by weight of N- (1-naphthyl) ethylenediamine dihydrochloride dissolved in 5% by weight of phosphoric acid) It was added to nitric acid ions (NO ₃ ^-) was measured the production of, and the concentration was evaluated as IC _50. As a negative control group, a group not treated with the sample was used.

As a result, the IC ₅₀ was measured to be about 5.63 μM.

Meanwhile, the compounds of the present invention can be formulated into various forms depending on the purpose. Examples of formulations for the composition of the present invention are illustrated below.

&Lt; Formulation Example 1 > Preparation of convergent lotion

To the purified water was added butylene glycol, glycerin, betaine, citric acid and sodium citrate, and the mixture was stirred and dissolved. Then, a mixture of a flavor and polyoxyethylene (60) hardened castor oil dissolved in ethanol was added thereto with gentle stirring. The Inoramo of the present invention was added thereto, and the mixture was sufficiently stirred and aged in a storage room. The content of each component is shown in Table 3 below.

Raw material Content (w / w%) Inoramo 3.0 Butylene glycol 7.0 glycerin 5.0 Polyoxyethylene (60) Hardened castor oil 0.2 ethanol 4.0 Betaine 2.0 Beta Glucan 0.5 Niacinamide 2.0 Citric acid 0.02 Sodium citrate 0.06 Preservative a very small amount Spices a very small amount Purified water to: 100

< Formulation example  2> Manufacture of nutrition lotion

The inoramic, butylene glycol, glycerin, carboxyvinyl polymer, arginine, allantoin, preservative and purified water of the present invention were heated to 70 to 75 캜 with stirring. The mixture of squalane, butylene glycol dicaprylate / dicaprate, sorbitan stearate, polysorbate 60, glyceryl stearate, rice bran oil, cetyl alcohol and stearyl glycite Nate mixture was emulsified with a homogenizer. The mixture was cooled to about 45 ° C with stirring and added with perfume. The mixture was cooled to 30 ° C and aged to prepare a nutritious lotion. The content of each component is shown in Table 4 below.

Raw material Content (w / w%) Inoramo 4.0 Butylene glycol 8.0 glycerin 5.0 Squalane 8.0 Rice bran oil 2.0 Butylene glycol dicaprylate / dicaprate 5.0 Sorbitan stearate 1.5 Polysorbate 60 1.0 Glyceryl stearate S 0.5 Cetyl alcohol 2.0 Stearyl glycyrrhetinate 0.2 Allantoin 0.1 Sodium hyaluronate 0.2 Niacinamide 2.0 Carboxyvinyl polymer 0.1 Arginine 0.1 Preservative a very small amount Spices a very small amount Purified water to: 100

< Formulation example  3> Production of Essence

4-cholesterol and cholesterol are dissolved by stirring to 70 to 80 ° C, and then purified water, allantoin, glycerin, xanthan gum, beta-glucan, etc. are dissolved at 70 ° C And then added with Inoramo of the present invention. When the mixture was cooled to 45 ° C while stirring, the flavor was added to the mixture, and the mixture was cooled to 30 ° C and aged. The content of each component is shown in Table 5 below.

Raw material Content (w / w%) Inoramo 1.0 Allantoin 0.1 Polyglyceryl 2-oleate 1.5 Ceramide 0.7 Setares-4 1.2 cholesterol 1.5 Dicetyl phosphate 0.4 1,3BG 5.0 Beta Glucan 0.5 Concentrated glycerin 0.5 Carboxyvinyl polymer 0.2 Xanthan gum 0.2 antiseptic a very small amount Spices a very small amount Purified water to: 100

< Formulation example  4> Pharmaceutical composition whitening cream

The inoramic, butylene glycol, glycerin, betaglucan, betaine, white mushroom extract, polyglutamic acid, carboxyvinyl polymer, arginine, allantoin, preservative and purified water of the present invention were heated to 70-75 DEG C with stirring . To the mixture was added a mixture of squalane, sunflower seed oil, jojoba oil, sorbitan stearate, polysorbate 60, cetostearyl alcohol, beeswax, glyceryl stearate and cocoa butter, which had been heated and stirred at 75 to 80 ° C, Niacinamide, hydroquinone and flavor were added to the mixture while cooling with stirring at about 45 ° C., and the mixture was cooled to 30 ° C. and aged to prepare a nutrition lotion. The content of each component is shown in Table 6 below.

Raw material Content (w / w%) Inoramo 10.0 Butylene glycol 8.0 glycerin 5.0 Squalane 10.0 Grape seed oil 3.0 Butylene glycol dicaprylate / dicaprate 5.0 Sorbitan stearate 1.5 Polysorbate 60 1.0 Glyceryl stearate 0.5 Stearyl glycinate 0.2 Cocoa butter 2.0 Beads wax 1.0 Cetostearyl alcohol 3.0 Hydroquinone 0.5 Allantoin 0.1 Sodium hyaluronate 0.2 Niacinamide 2.0 Carboxyvinyl polymer 0.1 Arginine 0.1 Preservative a very small amount Spices a very small amount Purified water to: 100

While the invention has been shown and described with reference to certain preferred embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the above description does not limit the scope of the present invention, which is defined by the limitations of the following claims.

Claims (8)

A skin lightening agent comprising a compound represented by the following formula (1).
[Chemical Formula 1]

The method according to claim 1,
Wherein said compound is separated from the shelf. The method according to claim 1,
Wherein said compound is a skin whitening agent for inhibiting melanin synthesis. A cosmetic composition for skin whitening comprising a compound represented by the following formula (1).
[Chemical Formula 1]

delete delete delete a) extracting an egg yolk with ethanol to obtain an ethanol extract; And
b) sequentially adding hexane, chloroform and ethyl acetate to the ethanol extract and fractionating to obtain a chloroform fraction containing a compound represented by the following Formula 1;
Which is extracted from a method for extracting an active ingredient for skin whitening.
[Chemical Formula 1]