KR20110075995A - Skin whitening composition containing gastrodin and glucosidase as active ingredients - Google Patents

Abstract

PURPOSE: A composition containing gastrodin and glucosidase for skin whitening is provided to ensure high whitening effect and to minimize side effects by formulation unstability and skin irritation. CONSTITUTION: A composition for skin whitening contains gastrodin and glucosidase as an active ingredient. Gastrodin is an alpha-gastrodin prepared by binding 1-8 alpha-D-glucoses by transglycosylation reaction using beta-gastrodin of chemical formula 1 or cyclodextrin glycosyltransferase. The gastrodin content is 0.01-10.0 weight% based on total composition. The glucosidase content is 75-150 units based on 3 weight% of gastrodin. The beta-gastrodin is a natural gatrodin or synthetic gastrodin. The natural gastrodin is isolated from Gastrodiae Rhizoma.

Description

Skin whitening composition containing gastrodin and glucosidase as active ingredients}

The present invention relates to a composition for skin whitening containing gastrodin and glucosidase, specifically, to beta-guestrodine or cyclodextrin glycosyltransferase of the formula (1) (cyclodextrin glycosyltransferase) From the used transglycosylation reaction, alpha-getrodine, in which 1 to 8 alpha-D-glucose is bound, and glucosidase, an enzyme that decomposes these guestodines into gastrodigenin and glucose. The present invention relates to a composition for skin whitening, which is contained in the same formulation or separated into different formulations and mixed immediately before use.

In general, when the skin is exposed to ultraviolet light, the epidermis becomes black, because the melanin, a black pigment, is synthesized and released from melanocytes. Tyrosinase of melanocytes synthesizes dopaquinone from tyrosine, and dopaquinone undergoes spontaneous oxidation to form melanin. Melanin absorbs excess light in the living body, but it is concentrated in specific areas to produce spots, blotch, etc., and may cause abnormal color fading of skin due to pregnancy blockage or skin pathology. In addition, many cosmetic compositions that are commercially available because of a strong desire to whiten the skin also contains a whitening agent component.

In order to prevent the skin from becoming black, a method of inhibiting melanin production is mainly used by inhibiting some reactions in the melanin production process, and as such a whitening agent, hydroquinone and its derivatives, arbutin and its derivatives, kojic acid and its derivatives Ascorbic acid is used. In addition, extracts of Aeggi bilberry (Application No. 2000-035398), Purified lettuce or apple (Registration No. 1998-147412) containing these ingredients have been used in whitening cosmetics. However, ascorbic acid, kojic acid, glutathione, lettuce extract, etc. have problems in skin irritation and stability of the formulation, its use is limited, and has a problem that must be used in a large amount for a long time. Furthermore, kojic acid and its derivatives are very expensive and difficult to use in mass-market products, and arbutin is browned by sunlight and heat, thereby inhibiting the inhibitory effect of tyrosinase and significantly less whitening effect than hydroquinone. Known.

Hydroquinone is the most effective whitening agent known to date and is widely used in prescriptions of non-prescription drugs or dermatologists, and there are a number of clinical reports showing it is effective in hyperpigmentation such as blemishes (Arndtet al., JAMA, 194: 965-967, 1965. Hydroquinone is a compound having a structure as shown in Formula 3, and acts as an inhibitor of tyrosinase, an enzyme that regulates the process of converting tyrosine to dopaquinone, which is an intermediate step of skin pigmentation in vivo.

However, hydroquinone has a disadvantage in that it is very unstable in the formulation, such as being highly reactive and easily discolored in aqueous solution or emulsifier and losing its efficacy. It can also cause chronic or acute skin irritation or contact skin allergy at low concentrations (Choudate et al., J. Ind. Med. 45 (6): 376-80, 1988). In order to avoid the side effects of such hydroquinone preparations, dermatology hospitals have been prescribed by adjusting the concentration of hydroquinone according to the skin sensitivity of the patient (Pearl E. et al., Archi. Dermatol. 131: 1453-1457, 1995).

Guest Lordine is the main active ingredient of Cheonma, which is known to have pharmacological effects such as analgesic, sedation, anticonvulsion, and central nervous system depression. The guest lodine present in nature is a beta-gustrodine in which beta-D-glucose is bound to the guest lodigenin, but the alpha-D- is derived from a transglycosylation reaction using a cyclodextrin glycosyltransferase. It is possible to prepare alpha-guestrodine in which 1 to 8 glucose is bound.

More specifically, beta-gustrodine is prepared by 1) chemical synthesis method, 2) cell culture method, and 3) purification from cheonma, and alpha-gestrodine is a sugar transfer reaction method using cyclodextrin glycosyltransferase. Can be synthesized from. In the case of synthesizing alpha-gestrodine by a sugar transfer reaction method, 1-8 glucose forms are mixed with guestlodigenin, and a single glucose molecule is prepared through a hydrolysis reaction. Since all of the chelodigenin glycosides that can be prepared using such a sugar transfer reaction method can be hydrolyzed to gyrodigenin by glucosidase, alpha-gestolodine in the present invention is cyclodextrin glycosyltransferase It is defined to include all forms of the guestlodigenin glycosides that can be synthesized from the sugar transfer reaction method.

Arbutin, which has a structure similar to that of guestrodin, is widely used in cosmetics for whitening, but it is not well absorbed by the skin and has a very low whitening effect compared to hydroquinone, and thus, in the experiment of inhibiting the activity of tyrosinase, which mainly acts on melanin production, It is known to have an effect of only 1/1000 of quinone (Kazuhisa Maeda and Minoru Fukuda, Arbutin: Mechanism of its depigmenting action in human melanocyte culture.J. Pharm.Exp. Therap. 1996; 276 (2): 765- 769).

In order to promote the skin absorption of arbutin, a method of using a nitrogen-added surfactant is known (US Pat. No. 5,759,528), and other studies have been conducted to develop new derivatives having high whitening effects. There is no successful report. Therefore, arbutin is more stable and less skin irritant than hydroquinone, but has not been widely used as a whitening agent due to low skin absorption and low whitening effect.

As mentioned above, hydroquinone is known as the most effective whitening agent in terms of activity, but it has been exposed to various side effects due to its skin toxicity and stability problems, and thus has a high skin whitening effect such as hydroquinone and less skin irritation and improved stability. There has been an urgent need for the preparation of whitening agents.

The inventors of the present invention have taken only the formulation stability of the guest rodin and the efficacy of the guest rodinogenin, and have been researched to develop a whitening agent having a high skin whitening effect and a low skin irritation and a formulation formulation. It is found that the whitening agent using the glucosidase as an active ingredient has little skin irritation and is stable, and when applied to the skin, it produces a high whitening effect by producing the active ingredient guestlodigenin due to the action of the enzyme. The present invention was completed by revealing that it can be used in a cosmetic composition for whitening.

Accordingly, it is an object of the present invention to provide a new skin whitening composition which has a high whitening effect and little skin irritation and is also stable in formulation.

In order to achieve the above-described technical problem, the present invention provides a composition for skin whitening, comprising a guest rodin and glucosidase as an active ingredient.

In the composition for skin whitening according to the present invention, the guest Lordine is alpha-D-glucose from the transglycosylation reaction using a beta-guestrodine or cyclodextrin glycosyltransferase of the formula (1) Is preferably 1-8 bonded alpha-guestine.

[Formula 1]

In the composition for skin whitening according to the present invention, the guest loadin content is 0.01 ~ 10.0% by weight based on the total composition content, the glucosidase content is 75 ~ 150 units based on the 3% by weight guest guest content desirable.

In the composition for skin whitening according to the present invention, the beta-gestrodine is preferably natural guestodine or synthetic guestodine, and in particular, natural beta-ketrodine may be prepared by extracting from cheonma.

In the composition for skin whitening according to the present invention, it is preferable that the guestidine and glucosidase are provided in separate formulations and mixed and used immediately before being applied to the skin.

In the composition for skin whitening according to the present invention, the reaction is carried out until one of the above guestidines or glucosidase is applied to the skin in the form of microcapsules, particulates or lipid vesicle dispersions according to techniques generally used in cosmetics. It is desirable to suppress.

The composition for skin whitening according to the present invention exhibits high skin whitening effect and low skin irritation and formulation stability by containing the guest rodin and the glucosidase which breaks down the guest rodin to produce the guest rodinogen. It can be used as an effective whitening agent.

In order to achieve the above object, the present invention relates to an alpha-D-glucose from a transglycosylation reaction using a guestodine, specifically, a beta-gustrodine or a cyclodextrin glycosyltransferase of the formula Is one to eight bound alpha-guestrodines and glucosidase, an enzyme that breaks down these gyrodins into gastrodigenin and glucose, are included in the same formulation or separated into different formulations. Provides a composition for skin whitening, characterized in that immediately before mixing to apply to the skin.

[Formula 1]

In the composition for skin whitening of the present invention, the guestodine content is 0.01 to 10.0% by weight, preferably 0.1 to 5.0% by weight based on the total composition. Glucosidase is preferably added in 75 to 150 units based on the 3% by weight of guestodine content. When the guestodine or glucosidase content is too low, the whitening effect intended in the present invention is not sufficiently exerted, and when too large, skin irritation may occur.

In the above, guestodine may be any of natural guestodine, synthetic guestodine, or any of the guestodine produced by a biotechnological method, and more preferably, an extract of a plant known to have a high guestodine content, such as cheonma, in the case of natural guestodine It may be obtained from or included in the composition of the present invention as the extract itself.

Glucosidase is an enzyme that catalyzes the production of sugars and aglycones by hydrolyzing glucosides, and is widely distributed in yeast, bacteria, fungi or internal organs of animals. Glucosidase in the composition for skin whitening of the present invention is not limited to enzyme sources such as those produced by gene recombination or isolated from natural products. In addition, the glucosidase may be added in the form of extracts of plants or microorganisms containing the enzyme, preferably microorganisms including aspergillus niger or plants including almonds, barley, oats, and the like. It can be used as an extract form of.

Of course, it is not limited to those listed above, and any glucosidase capable of hydrolyzing the guestrodin to produce the guestlodigenin can be used in the composition for skin whitening of the present invention regardless of its origin. It will be apparent to those skilled in the art to which the present invention pertains.

In the composition for skin whitening of the present invention, guestrodin and glucosidase may be provided as separate formulations, respectively, and the two components may be mixed and used immediately before application to the skin. In the case of using the guest Lordin and glucosidase at the same time, the guest Lordin is broken down into the guestidinine and glucose under the influence of glucosidase. The whitening effect is shown by the guest rhodigenin produced by the reaction, compared to the existing product consisting of hydroquinone, it can be obtained an excellent effect in stability and skin hypoallergenicity.

The composition for skin whitening of the present invention may be used in addition to flexible skin lotion (skin), nutrition lotion (milk lotion), nutrition cream, massage cream, essence, pack, etc., but is not limited thereto. In addition, other whitening agents, humectants, antioxidants, ultraviolet absorbers, surfactants, thickeners, alcohols, preservatives, gelling agents, fragrances, fillers, dyes and the like used in ordinary cosmetics may be appropriately blended as necessary. The amount of the adjuvant is appropriately used in the cosmetic field, such as about 0.01 to 20% of the total weight of the composition.

The skin whitening composition of the present invention is in any pharmaceutical form commonly used for topical application, for example in the form of dispersions, emulsions, microcapsules, particulates or vesicle dispersions in the form of solutions, gels, lotions or emulsions. Can be. Particularly when incorporation of guestrodin and glucosidase into the same formulation, applying one of the guestrodin or glucosidase to the skin in the form of microcapsules, particulates or lipid vesicle dispersions according to techniques commonly used in cosmetics You can also maximize stability by suppressing the reaction until now.

In a preferred embodiment of the present invention, high performance liquid chromatography and gas chromatograph-mass spectrometry are widely used for qualitative and quantitative analysis of organic substances. It was confirmed that guestrodin was completely decomposed into gyrodigenin and glucose.

In addition, in order to confirm whether the mixed formulation of the guestlodigenin and glucosidase of the present invention is more stable than the hydroquinone alone formulation, the mixture is reacted at 37 ° C for 10 minutes, and then cooled on ice to stop the reaction, and the glucosidase After removing the enzyme activity of the absorbance at 400 nm by measuring the browning degree of each sample was determined as a ratio to the degree of discoloration of hydroquinone. As a result, the mixed formulation of guestodine and glucosidase had a very low discoloration level compared to the hydroquinone alone formulation, and thus the stability in the formulation was excellent. In particular, the lower the glucosidase concentration, the higher the stability. This is because the degradation of guestodine is increased in proportion to the concentration of glucosidase.

Meanwhile, in another embodiment of the present invention, the degree of primary stimulation of the gyrodigenin produced by glucosidase in the human body was measured by using a closed patch test method in healthy adult men and women. As a result, unlike the control group directly patched with hydroquinone, it was found that the primary irritation was hardly observed in the skin of the experimental group that was coated with the mixed solution of the guestlodine and glucosidase.

In addition, by irradiating the skin with UV light on healthy adults and men and women, the pigment is deposited on the skin, and then a combination of the preparation of the first agent containing the guest rodin and the second agent containing the glucosidase is used simultaneously. As a result of investigating the whitening effect by sidase, the whitening agent composition of the present invention showed a superior whitening effect than when using only guestidine, and did not show side effects due to skin irritation when hydroquinone was treated.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example 1 Identification of the Decomposition Product of Gerosidine by Glucosidase

0.05% guestrodin (Sigma, USA) and 20 units / ml beta [1-4] glucosidase (Sigma, USA, EC 3.2.1.21) in 0.1 M sodium acetate buffer (pH 5.0) for 30 minutes at 37 ° C. Reacted. In order to separate and confirm the product after the reaction, the guestrodin reaction solution and the control material (gestrodine, guestodigenin) were mixed with ethanol, and filtered through a 0.45 μm membrane to perform high performance liquid chromatography analysis. In order to analyze the cherodidigenin from the glucose reaction solution of the guest Lordine, it was extracted with acetylacetate and trimethylsilylation, and the resulting glucose was TMS derivatized from the residue after the extraction of the ethyl acetate. The TMS derivatives were analyzed by GC-MS to identify gyrodigenin and glucose from the beta [1-4] glucosidase reaction solution of gyrodine.

1 is a result showing the production of the guest lodigenin after the reaction of the guest gludinid and glucosidase, it can be seen that the conversion of the guest rodin hydrolyzed by the glucosidase is completely converted to the guest lodigenin.

2 and 3 are more qualitative results of the production of gyrodigenin and glucose after the reaction of gyrhodin and glucosidase, from the results of mass spectrometry to conversion of gyrhodidine to gyrhodidigenin and glucose You can see that.

Example 2 Investigation of Stability of Mixed Formulations

The degree of discoloration was examined to determine whether the mixed formulation of the guestlodine and glucosidase of the present invention is more stable than the hydroquinone alone formulation. First, a solution of 1, 10, 20 units / ml of 100 mM guestodine and glucosidase, respectively, was reacted at 37 ° C. for 10 minutes in 1 ml of sodium acetate buffer, and then cooled on ice to stop the reaction. In order to remove the enzyme activity of the glucosidase, 1 ml of cold 20% TCA (trichloroacetic acid) solution was mixed and centrifuged (12,000 rpm) to remove the glucosidase in the solution. After leaving each solution at 37 ° C. for 24 hours, the absorbance at 400 nm was measured to determine the browning degree of each sample as a ratio to the degree of discoloration of the hydroquinone solution. As a control, a single formulation of 100 mM guestodine and 100 mM hydroquinone (Sigma, USA) was used.

As can be seen from the results of Table 1, the mixed formulation of guestodine and glucosidase has a very low discoloration level compared to the hydroquinone alone formulation, so that the stability in the formulation is excellent. In particular, the lower the glucosidase concentration, the more stable the formulation. It can be seen that high. This is because the degradation of guestodine is increased in proportion to the concentration of glucosidase.

Kinds Absorbance (400 nm) Discoloration degree (% hydroquinone discoloration) Buffer 0.002 0.6 Hydroquinone solution 0.356 100 Guest Rodin 0.003 0.8 Veyrodin + 1 μl glucosidase 0.008 2.3 Veyrodin + 10 μl glucosidase 0.024 3.4 Veyrodin + 20 μl glucosidase 0.085 4.2

* The degree of discoloration was compared with the hydroquinone solution discoloration absorbance based on 100%.

Example 3 Inhibition of Primary Stimulation of Skin by Cyclodinate and Glucosidase Reaction

Thirty healthy men and women were selected and a closed patch test was performed to investigate the degree of skin irritation of the mixed solution of guestodine and glucosidase. First, the solution of Example 2 was patched to the lower arm of the test subject for 24 hours, and the skin was observed for 30 to 48 hours after removal to classify the degree of stimulation according to the criteria of Table 2.

As can be seen from Table 3, the solution of hydroquinone only showed a very strong reaction, whereas the mixed solution of guestidine and glucosidase showed no primary irritation on the skin.

sign Criteria ? + Suspicious reaction, slight erythema + Weak reactions (without vesicles), erythema, papules ++ Strong reactions (with vesicles), erythema, papules, vesicles +++ Very strong positive reaction, alveolar reaction --- voice

Test substance
Responsiveness 24 hours 48 hours Buffer - - Hydroquinone solution +++ +++ Guest Rodin - - Veyrodin + 1 μl glucosidase - - Veyrodin + 10 μl glucosidase - - Veyrodin + 20 μl glucosidase - -

Example 4 Skin Whitening Effect of the Gyrhodine and Glucosidase Blend Formulations

Practical formulations were prepared based on the above examples to investigate the skin whitening effect of the guestrodin and glucosidase reactions.

As described below, the first and second agents containing glucosidase are prepared separately, and then the first agent and the second agent are mixed in the palm in a ratio of 1: 1 immediately before use. Formulations were prepared to compare the whitening effect with comparative formulations in which only one type of guestodine or hydroquinone was added.

For the 10 healthy men and women, a pair of 4 holes with a diameter of 1.5 cm was attached to the lower arm of both arms, and the UV light (Philips TL20W / 12UV lamp and TM02 / 09UV lamp) was applied at a distance of 10 cm. Skin pigments were deposited by MED twice a day for 2 days. The first and second agents of Table 4 were mixed and used at a ratio of 1: 1, which was used as the experimental group. The effect was observed visually.

As a result, as shown in Table 5, there was almost no effective whitening effect in the case of not including the whitening component or only in the case of the guest rodin. It was found to be not suitable for use because of its high height. In contrast, when used in combination with guestidine and glucosidase, not only showed a high whitening effect but also no skin side effects.

Ingredient (Unit: weight%) The first 2nd Comparative Example 1 Comparative Example 2 Comparative Example 3 Guest Rodin 3.0 - - 3.0 - Glucosidase - 100 unit - - - Hydroquinone - - - - 2.0 glycerin 10.0 1.0 10.0 10.0 10.0 Propylene glycol 5.0 0.5 5.0 5.0 5.0 Cellulose gum 0.3 0.03 0.3 0.3 0.3 Hyaluronic acid extract 10.0 1.0 10.0 10.0 10.0 pH regulator Quantity Quantity Quantity Quantity Quantity Purified water to 100 to 10 to 100 to 100 to 100

Test Product Skin pigmentation improvement effect Remarks Experimental Example 1 6 3 One - Valid effect Comparative Example 1 5 2 8 4 Slightly effect Comparative Example 2 6 3 7 5 No change Comparative Example 3 5 2 - 3 Skin side effects

Preferred preparations of the essence using the skin whitening composition containing the guest of the present invention and glucosidase are as follows.

* First agent: Second agent = 1: 1 Mix in the palm and apply to the face before use in a volume ratio.

Preparation Example 1 Preparation of Essence Using Composition for Skin Whitening of the Present Invention

The first 2nd ingredient Content (% by weight) ingredient Content (% by weight) Guest Rodin 3.0 Glucosidase 100 units glycerin 10.0 glycerin 1.0 Propylene glycol 5.0 Propylene glycol 0.5 Cellulose gum 0.3 Cellulose gum 0.03 Hyaluronic acid extract 10.0 Hyaluronic acid extract 1.0 pH regulator Quantity pH regulator Quantity incense a very small amount incense a very small amount antiseptic a very small amount antiseptic a very small amount Pigment a very small amount Pigment a very small amount Purified water to 100 Purified water to 10

<Preparation Example 2> Preparation of nutritional lotion using the composition for skin whitening of the present invention

The first 2nd ingredient Content (% by weight) ingredient Content (% by weight) Guest Rodin 3.0 Glucosidase 100 units Beeswax 3.0 Beeswax 0.3 Floating paraffin 4.0 Floating paraffin 0.4 glycerin 10.0 glycerin 1.0 Carboxy Vinyl Polymer 0.1 Carboxy Vinyl Polymer 0.01 Polysorbate 60 1.1 Polysorbate 60 0.11 Propylene glycol 5.0 Propylene glycol 0.5 pH regulator Quantity pH regulator Quantity Incense, preservative, coloring a very small amount Incense, preservative, coloring a very small amount Purified water to 100 Purified water to 10

Preparation Example 3 Preparation of Pack Using Skin Whitening Composition of the Present Invention

The first 2nd ingredient Content (% by weight) ingredient Content (% by weight) Guest Rodin 3.0 Glucosidase 100 units Polyvinyl alcohol 14.0 Polyvinyl alcohol 1.4 glycerin 10.0 glycerin 1.0 Cellulose gum 0.3 Cellulose gum 0.03 PEG 4000 1.0 PEG 4000 0.1 Propylene glycol 5.0 Propylene glycol 0.5 pH regulator Quantity pH regulator Quantity Incense, pigment, preservative a very small amount Incense, pigment, preservative a very small amount Purified water to 100 Purified water to 10

<Example 4> Preparation of nutrition cream using the composition for skin whitening of the present invention

The first 2nd ingredient Content (% by weight) ingredient Content (% by weight) Guest Rodin 3.0 Glucosidase 100 units Floating paraffin 10.0 Floating paraffin 1.0 Propylene glycol 5.0 Propylene glycol 0.5 Beeswax 7.0 Beeswax 0.7 glycerin 10.0 glycerin 1.0 Polysorbate 60 1.3 Polysorbate 60 0.13 pH regulator Quantity pH regulator Quantity Incense, preservative, coloring a very small amount Incense, preservative, coloring a very small amount Purified water to 100 Purified water to 10

1 is a diagram showing a high performance liquid chromatogram of an enzyme reaction final product after treating glucosidase to guest rodin,

Figure 2 is a diagram showing the results of analysis by gas chromatograph-mass spectroscopy after TMS derivatization of the guest lodigenin produced from the enzymatic reaction end obtained after treatment with glucosidase in the guest rodin,

Figure 3 is a diagram showing the results of analysis by gas chromatograph-mass spectroscopy after TMS derivatization of the glucose produced from the enzymatic reaction end obtained after treatment with glucosidase in the guest rodin.

Claims (7)

A composition for skin whitening, comprising a guest rodin and a glucosidase as an active ingredient. The method according to claim 1, wherein the guest lordine is 1 to 8 alpha-D-glucose from the transglycosylation reaction using a beta-guestrodine or cyclodextrin glycosyltransferase of the formula (1) A composition for skin whitening, characterized in that it is bound alpha-gustrodine. [Formula 1]

The method according to claim 1 or 2, wherein the guest Lordin content is 0.01 to 10.0% by weight based on the total composition content, the glucosidase content is 75 to 150 units based on the 3% by weight guestrodine content Skin whitening composition. The composition for skin whitening according to claim 2, wherein the beta-guestrodine is natural guestodine or synthetic guestodine. The composition for skin whitening according to claim 4, wherein the natural guestodine is prepared by extracting from cheon-ma. The composition for skin whitening according to claim 1 or 2, wherein the guestodine and glucosidase are each provided in separate formulations and mixed immediately before being applied to the skin. The method according to claim 1 or 2, wherein the component of said guestodine or glucosidase is suppressed until it is applied to the skin in the form of microcapsules, particulates or lipid vesicle dispersions according to techniques generally used in cosmetics. Skin whitening composition, characterized in that.

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