WO2009107896A1

WO2009107896A1 - Compositions containing kaempferia pandurata extract for preventing or treatment skin disease

Info

Publication number: WO2009107896A1
Application number: PCT/KR2008/002006
Authority: WO
Inventors: Jae Kwan Hwang; Chan Woo Lee; Han Sung Kim; Il Young Kwack; Xian Li Song; Jin Wong Kim; Han Kon Kim; Jae Seok Shim
Original assignee: Amorepacific Corporation
Priority date: 2008-02-29
Filing date: 2008-04-10
Publication date: 2009-09-03
Also published as: KR20090093477A

Abstract

The present invention relates to a composition effective in preventing or treating skin diseases, more particularly to a composition comprising Kaempferia pandurata extract and effective in preventing or treating skin diseases associated with sebum and skin diseases caused by skin irritation or inflammation. The composition comprising the Kaempferia pandurata extract is effective in preventing or treating skin diseases, for example, skin diseases associated with sebum and skin diseases caused by skin irritation or inflammation.

Description

[DESCRIPTION] [Invention Title]

COMPOSITIONS CONTAINING KAEMPFERIA PANDURATA EXTRACT FOR PREVENTING OR TREATMENT SKIN DISEASE

[Technical Field]

The present invention relates to a composition effective in preventing or treating skin diseases, more particularly to a composition comprising Kaempferia pandurata extract and effective in preventing or treating skin diseases associated with sebum and skin diseases caused by skin irritation or inflammation.

[Background Art]

In general, sebum is produced to protect hair and skin from bacteria and keep them from drying. However, excessive sebum secretion may result in such skin problems as greasy skin, loose makeup, enlarged skin pores and acne proneness after the puberty, and the like.

Sebum is secreted in the sebaceous gland positioned in the pi losebaceous duct of skin, and reaches the skin surface through the hair follicle duct. Excessive secretion of sebum is related with a variety of factors. Particularly, the activation of sebaceous gland cells due to hormonal action is known to be the most important factor.

Excessive secretion of sebum is a matter of concern in the field of skin care and cosmetics, because it causes loose makeup or enlargement skin pores. Nevertheless, active studies have not been performed yet. Cosmetics for suppressing sebum that are currently used comprise porous powder for temporarily adsorbing sebum or contain medicinal herb extract, such as ivy extract, known to suppress sebum. However, such cosmetics cannot provide a sufficient effect.

Peroxisome proliferator-activated receptors (PPARs) are known nuclear hormone receptors having the three subtypes of α , J3 / δ and Y . PPAR- α has been identified first based on the mechanism of controlling genes encoding fatty acid oxidase by the reaction with a peroxisome proliferator factor such as a fibric acid derivative. In some literatures, it is disclosed that fatty acids play an important role in tissues expressed by PPAR- α . Lipid activators of PPAR- α , such as linoleic acid, etc., are known to those skilled in the art. It is demonstrated that such activators enhance formation of an epithelial barrier in vitro.

Also, it has been found that PPAR-γ activators increase insulin sensitivity and decrease glucose and insulin in a recent animal model test. This indicates that PPAR- y may be applied to treatment of diabetes mellitus or a par of complications thereof. Additionally, it is reported that PPAR-γ affects maturation of sebocyte in the sebaceous gland. Recently, such effect of PPAR-γ has been applied in the cosmetic field. For example, in Korean Patent Application Nos. 2007-0070303 and 2007-0028902 by the inventors of the present invention, the effect of kaempferol suppressing sebum by decreasing the activity of PPAR-y is mentioned in part. However, studies using Kaempferia pandurata extract have not been reported yet.

Since the stratum corneum of human skin is covered by the sebum membrane which is formed naturally from the sebum secreted from the sebaceous gland and the moisture from the sweat gland, it is maintained in smooth, glossy and moist status. The skin condition is ideal when the moisture content of the stratum corneum is about 20%. When the moisture content is below 10%, the horny layer is sloughed off easily, making the skin vulnerable to infiltration of foreign matters, and inflammations and allergic reactions associated therewith. In particular, such inflammations and allergic reactions become severe in aged people, whose skin metabolism is not so effective.

Further, skin tends to be more sensitive due to various external stimulations, including dry indoor environment, pollution, ultraviolet (UV) rays, wind, misused cosmetics or detergents, and the like. Such sensitized skin may experience visible symptoms such as erythema, and, in severe cases, inflammatory responses accompanied by itching, irritation, etc.

Especially, although cosmetics are used to protect, beautify and clean the skin, at the same time, they may induce adverse reactions when applied on the skin. It is because some components without regard to the purpose of skin protection are added for the manufacture of cosmetics. For example, so1 Libi lizer , emulsif ier , preservative, fragrance, UV blocker, and other additives are used in the manufacture of cosmetics in order to provide various effects. These components may induce skin dryness, erythema, skin hardening, itching, glowing, and the like, depending on individuals.

Recently, various studies are carried out in order to minimize the adverse reactions caused by the components added to cosmetics. There have been attempts to alleviate immune responses through cytokine and inflammatory mediators, and moderate skin irritation by strengthening skin barrier and protecting skin. Specific examples include the use of bradykinin antagonist for control of inflammatory mediators and adverse reactions of sensitive skin, the use of TNF- α inhibitor, one of cytokines, for treatment of sensitive skin, and the like.

Human kerat inocytes or Langerhans' cells can produce and secrete various cytokines by the exterior material. These cytokines promote and activate the immune system and are essential in stimulation of skin or topical inflammatory response. Actually, increase of inter leukin-6 ( IL-6) , inter leukin-1 (IL-I) and tumor necrosis factor- α (TNF- α )has been reported in irritant contact dermatitis. Intercellular cell adhesion mo1 ecuIe-I (ICAM-I) is a molecule essential in the adhesion of kerat inocytes to lymphocytes. In stimulated skin irritation or topical inflammatory response, increase of ICAM-I expression in the level of proteins and genes is observed. Since the expression of ICAM-I on the surface of kerat inocytes is induced by interferon- Y (IFN- Y ), an inflammatory cytokine, and functions to reduce irritation, it is expected to be applied to suppress the inflammatory adverse responses .

Currently available chemicals that can be used to alleviate irritations or inflammations such as erythema and edema are as follows. Non-steroidal substances include flufenamic acid, ibuprofen, benzydamine, indomethacin, and the like, and steroidal substances include prednisolone, dexamethasone, and the like. Besides, allantoin, azulene, ε -aminocaproic acid, hydrocortisone (v), glycyrrhizic acid and its derivatives (β-glycyrrhizic acid and others), and the like are known to have ant i-inflammatory effects.

However, because cosmetics are always associated with the problem of skin irritation, studies have been actively carried out to solve this problem. Most of the anti-inflammatory agents known thus far are restricted in use for cosmetics, because they have problems with respect to skin safety or product stability. For example, the ant i-inflammatory agent indomethacin cannot be used in cosmetics, use of hydrocortisone is restricted in quantity, and use of glycyrrhizic acid and its derivatives is restricted in concentration because of stabilization or solubility problem.

Recently, research and development of compositions comprising various natural ingredients that can reduce skin irritation and inflammation and cosmetics using them are carried out actively in order to meets the consumers' needs.

[Disclosure]

[Technical Problem]

The inventors of the present invention found out that a composition comprising Kaempferia pandurata extract as active ingredient and a cosmetic composition comprising the same are effective in preventing or treating skin diseases, and completed the present invention.

[Technical Solution]

The present invention provides a composition for preventing or treating skin diseases, which comprises Kaempferia pandurata extract as active ingredient, and a cosmetic composition comprising the same. [Description of Drawings]

FIG. 1 shows the images of histopathological slides treated with Kaempferia pandurata extract according to the present invention.

[Best Mode]

Accordingly, the composition for preventing or treating skin diseases according to the present invention comprises Kaempferia pandurata extract as active ingredient.

Kaempferia pandurata deseribed herein is as following.

Kaempferia pandurata is a medicinal plant belonging to the Zingiberaceae family and grows naturally in Southeast Asia. Its root is used to treat cold, enteritis, skin disease and urethral pain. Pinocembrin chalcone, cardamonin, pinocembrin, pinostrobin, 4-hydropandurat in A, panduratin A, etc. included in Kaempferia pandurata are reported to have anticancer effect, and the flavonoid-based dihydrochalcone compound is known to have insecticidal effect.

However, up to now, there is no case of alleviating skin irritation and inflammation by inhibiting the expression of inflammatory cytokine and reducing the activity of PPAR- y and thereby suppressing sebum, using Kaempferia pandurata.

The composition comprising Kaempferia pandurata extract according to the present invention is effective in preventing or treating skin diseases, including both sebum-related diseases and skin diseases caused by skin irritation or inflammation.

First, with respect to sebum-related diseases, the Kaempferia pandurata extract according to the present invention is effective in suppressing the secretion of sebum. It is because the Kaempferia pandurata extract reduces the activity of peroxisome proliferator-activated receptor- y (PPAR- Y ) and inhibits the generation of sebum.

The sebum-related diseases include acne, hair loss, seborrheic dermatitis, and various other skin diseases caused by over-secretion of sebum.

With respect to skin diseases caused by skin irritation or inflammation, the Kaempferia pandurata extract is effective in inhibiting the expression of at least one inflammatory mediators selected from inflammatory cytokines IL-I α IL-6 and IL-8, and MCP-I (monocyte chemotactic protein-1). By way of this, it inhibits the formation of erythema or edema and provides the effect of treating skin diseases caused by skin irritation or inflammation.

In an embodiment of the present invention, the Kaempferia pandurata extract may be obtained from an organic solvent extract or oil pressed from Kaempferia pandurata.

For the solvent, purified water, methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, petroleum ether or a mixture thereof may be used. Preferably, ethanol may be used. In addition, any solvent adequate to extract Kaempferia pandurata as active ingredient may be used without particular limitation.

The present invention also provides a cosmetic composition using the aforesaid composition. Because the cosmetic composition comprises the Kaempferia pandurata extract according to the present invention, it is effective in preventing or treating skin diseases through suppressing sebum generation and alleviating skin irritation and inflammation.

In a preferred embodiment, the Kaempferia pandurata extract is comprised in an amount of 0.001 to 20 weight %, based on the total weight of the cosmetic composition. When the content of the Kaempferia pandurata extract is below 0.001 weight %, the expected effect of preventing or treating skin diseases may not be attained. And, when the content exceeds 20 weight %, it is difficult to obtain a stable composition. More preferably, the Kaempferia pandurata extract is comprised in an amount of 0.1 to 10 weight %, based on the total weight of the cosmetic composition.

The cosmetic composition according to the present invention can be prepared into various formulations.

In certain embodiments, the cosmetic composition may be a skin adhesion type cosmetic composition, such as softening toner, astringent toner, nourishing toner, nourishing cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, body lotion, body cream, body oil, body essence, makeup base, hair dye, shampoo, rinse or body cleaner.

In other preferred embodiments, the cosmetic composition may be a transdermal type cosmetic composition, such as lotion, ointment, gel, cream, patch or spray.

[Mode for Carrying Out the Invention]

The present invention is further illustrated by the following examples. However, the following examples are only exemplary and for understanding of the present invention, and the present invention is not limited by them.

In the following Test Examples, activity of PPAR- γ was measured using the Kaempferia pandurata extract according to the present invention (Test Example 1), and the effect of inhibiting the generation of sebum was tested (Test Example 2). Further, the degree of the control in expression of inflammatory mediators in sodium lauryl sulfate (SLS)-stimulated cells was tested using the Kaempferia pandurata extract (Test Examples 3 and 4). At last, irritation test was performed on ICR mice in order to evaluate the effect of alleviating skin irritation in vivo (Test Example 5).

[Test Example 1] Determination of PPAR- y activation capability by one- factor-at-a-time (OFAT) method

CV-I cells (ATCC CCL 70), i.e., monkey kidney epithelial cell line, were subcultured in a DMEM (Dulbecco's modified Eagle medium) medium containing 10% fetal bovine serum treated with charcoal /dextrin. A phenol red-free medium was used to avoid the effect of estrogen upon phenol red. As O

plasmids, used were plasmids having PPRE (PPARs responsive element) as promoter, being activated by PPAR- y (gene-containing PPAR- y and ligand-bound PPAR- Y ) bound next to the universal promoter expressed under general culture conditions, having firefly luciferase genes to be a reporter afterwards, and a plasmid to which renilla luciferase genes were bound as a reference.

4

CV-I cells were plated on a 24-well plate at a concentration of 5x10 cells per well and cultured for 24 hours. Then, the above three types of plasmid genes were subjected to transient transfection by using a TransFast Kit available from Promega Co. After culturing for 24 hours, the cultured product was washed with IxPBS (phosphate buffered saline), treated with a suitable concentration of sample materials, cultured again for 24 hours, and washed with IxPBS two times. Next, the cells were lysed with IxPLB (passive lysis buffer, Promega), and determined for luciferase activity by using a Dual-Luciferase Reporter Assay System Kit (Promega) in the samples and controls.

In this test example, as a positive control, troglitazone known as a PPAR- _Y ligand was used. As negative controls, DMSO used to dissolve the samples and non-treated group were used. Each reported value was calculated as a ratio based on the negative control. The results are shown in the following Table 1, wherein each value is the mean value obtained after carrying out the test three times.

[Table 1]

As can be seen from Table 1, the Kaempferia pandurata extract shows an excellent effect of inhibiting PPAR- y activity.

[Test Example 2] Determination of effect of alleviating acne conditions by suppressing sebum generation

To determine the effect of suppressing sebum generation and alleviating acne conditions of the composition according to the present invention, two groups including 26 men and women aged 14-28 years were allowed to use the lotion as described in the following Table 2 for 4 weeks in a conventional manner. The effect of improving skin conditions was determined according to the evaluation criteria as described in Table 3 below based on the comments of the users. The evaluation results are shown in Table 3. [Table 2]

[Table 3]

As can be seen from Table 3, the composition comprising Kaempferia pandurata extract according to the present invention shows a higher effect of suppressing sebum generation and alleviating acne conditions.

[Test Example 3] Confirmation of effect of inhibiting biosynthesis of SLS- stimulated IL-I α in HaCaT keratinocyte cell line

Using a DMEM medium (purchased from Lonza, USA) containing 10% fetal bovine serum (FBS, purchased from Gibco, USA) and 1% penicillin-streptomycin (purchased from Gibco, USA), keratinocytes (HaCaT cell line, purchased from Dr. N.E. Fusenig, Deutsches Krebsforschungszentrum, Heidelberg, Germany) to

4 be tested were plated on a 48-well plate at a concentration of 2x10 cells per well and cultured for 24 hours under the condition of 37^°C and 5% CO2. After removing the culture medium, washing with 200 μi of PBS once, and adding 200 μi of a DMEM medium not containing FBS, the cells were cultured for 24 hours. Then, the sample extracts were treated with a DMEM containing 1% FBS at a concentration as described in Table 4 below for 10 minutes. After treating with iμM SLS (Sodium Lauryl Sulfate: purchased from Sigma-Aldrich, USA), culturing was carried out for 24 hours. 50 μi of the culture medium was taken and reacted for 4 hours with a 96-well plate pre~coated with mouse anti-human IL-I α (purchased from Pharmingen, USA), for 1 hour with mouse biotinylated anti-human IL-I α (purchased from Pharmingen, USA), and subsequently for 30 minutes with avidin-horseradish peroxidase conjugate (purchased from Pharmingen, USA), by ELISA.

The results were confirmed by treating the samples with 2,2'-amino- bis(3-ethylbenzthiazoline-6-sulfonic acid) (purchased from Sigma-Aldrich, USA) for 30 minutes in shade and measuring absorbance at 405 nm. The relationship between the absorbance and inhibition capability was obtained from the standard curve of the standard material rh IL-I α (purchased from eBioscience, USA). The secretion amount of IL-I α was calculated by substituting for the absorbance of the sample. The alleviation of irritation was calculated by the following Equation 1. [Equation 1]

Alleviation of irritation (%) = 100 x [l-(Secretion amount of IL-I α in sample-treated group / Secretion amount of IL-I α in SLS-treated group)] [Table 4]

[Test Example 4] Confirmation of effect of inhibiting biosynthesis of SLS- stimulated IL-4, IL-8 and CMP-I in HaCaT keratinocyte cell line

Using a DMEM medium (purchased from Lonza, USA) containing 10% FBS (purchased from Gibco, USA) and 1% penicillin-streptomycin (purchased from Gibco, USA), keratinocytes (HaCaT cell line, purchased from Dr. N.E. Fusenig, Deutsches Krebsforschungszentrum, Heidelberg, Germany) were plated on a 48-

4 well plate at a concentration of 2x10 cells per well and cultured for 24 hours under the condition of 37°C and 5% CO2. After removing the culture medium, washing with 200 μi of PBS once, and adding 200 μi of a DMEM medium not containing FBS, the cells were cultured for 24 hours.

Then, the sample extracts were treated with a DMEM containing 1% FBS at a concentration as described in Tables 5 to 7 below for 10 minutes. After treating with 1 μM SLS (purchased from Sigma-Aldrich, USA), culturing was carried out for 24 hours. 50 μi of the culture medium was taken and the effect of inhibiting release of cytokine was evaluated using IL-4, IL-8 and CMP-I ELISA kits (purchased from BD Pharmingen, USA). The relationship between the absorbance and inhibition capability was obtained from the standard curve of the standard materials rh IL-4, rh IL-8 and rh CMP- Kpurchased from BD Pharmingen, USA). The secretion amount of cytokine was calculated by substituting for the absorbance of the sample. The alleviation of irritation was calculated by the following Equation 2. [Equation 2]

Alleviation of irritation (%) = 100 x [l-(Secretion amount of cytokine in sample-treated group / Secretion amount of cytokine in SLS-treated group)] [Table 5]

[Table 6]

[Table 7]

[Test Example 5] Evaluation of effect of alleviating irritations and inflammations in animal model

One hour before and six hours after applying the inflammation-indueing material TPA (^-O-tetradecanoylphorbol-lδ-acetate) on the right ear of an ICR mouse, 20 μi of the test material and the ant i-inflammatory agent hydrocortisone were respectively applied, as described in Table 5. TPA was applied on the right ears of all the mice in the relevant group at a dose of 25 βg/μi. 18 hours after TPA application, the right ears were cut using a 6 mm biopsy punch and weighed. The ear tissues were fixed in 10% neutral formalin and prepared into tissue section slides by embedding in paraffin after dehydration. The tissues were observed with an optical microscope after staining with H&E (hematoxylin & eosin).

Six mice were used per each group. The statistical difference of the weight of the ears of each group was analyzed by two-sample T-test with a confidence interval of 95%. The results are given in Table 8 below.

The histopathological slides were photographed (X200) using an optical microscope. The cross-sectional thickness was determined using the image system i-Solution Lite [IMT (Image and Microscope Technology) Corporation], and the change in the number of inflammatory cells was observed. The results are shown in FIG. 1. Following one-way ANOVA, the cross-sectional thickness was analyzed by the Tukey's method at a confidence interval of 95%. The inhibition ratio was calculated by the following Equation 3. [Equation 3]

% Inhibition = [(Average weight of ears treated with TPA only - Average weight of ears treated with test material) / (Average weight of ears treated with TPA only - Average weight of ears treated with hydrocortisone) x 100] [Table 8]

As can be seen from Table 8 and FIG. 1, the Kaempferia pandurata extract according to the present invention is effective in alleviating skin irritations.

[Industrial Applicability]

As can be seen from the foregoing, the composition comprising Kaempferia pandurata extract according to the present invention is effective in preventing or treating skin diseases, for examples, skin diseases associated with sebum and skin diseases caused by skin irritation or inflammation.

The present invention has been described in detail with reference to preferred embodiments thereof. However, it will be appreciated by those skilled in the art that changes may be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims

o

[CLAIMS] [Claim 1]

A composition for preventing or treating skin diseases, which comprises Kaempferia pandurata extract as active ingredient.

[Claim 2]

The composition for preventing or treating skin diseases according to claim 1, wherein the Kaempferia pandurata extract suppresses the secretion of sebum.

[Claim 3]

The composition for preventing or treating skin diseases according to claim 1, wherein the skin diseases are inflammatory skin diseases.

[Claim 4]

The composition for preventing or treating skin diseases according to claim 2, wherein the Kaempferia pandurata extract suppresses the secretion of sebum by reducing the activity of peroxisome proliferator-activated receptor- Y (PPAR- Y ) .

[Claim 5]

The composition for preventing or treating skin diseases according to claim 2, wherein the skin diseases are acne, hair loss or seborrheic dermatitis.

[Claim 6]

The composition for preventing or treating skin diseases according to claim 3, wherein the Kaempferia pandurata extract suppresses the expression of at least one inflammatory mediator selected from the group consisting of IL-I α (inter leukin-lα), IL-6 ( interleukin-6), IL-8 (inter leukin-8) and MCP- 1(Monocyte Chemotactic Protein-1).

[Claim 7]

The composition for preventing or treating skin diseases according to claim 3, wherein the Kaempferia pandurata extract suppresses the generation of erythema or edema.

[Claim 8]

The composition for preventing or treating skin diseases according to claim 1, wherein the Kaempferia pandurata extract is obtained from an organic solvent extract or oil pressed from Kaempferia pandurata.

[Claim 9]

The composition for preventing or treating skin diseases according to claim 8, wherein the organic solvent is ethanol .

[Claim 10]

The composition for preventing or treating skin diseases according to any of claims 1 to 9, wherein the composition is a cosmetic composition.

[Claim 11]

The composition for preventing or treating skin diseases according to claim 1, wherein the Kaempferia pandurata extract is comprised in an amount of 0.001 to 20 weight % based on the total weight of the composition.

[Claim 12]

The composition for preventing or treating skin diseases according to claim 11, wherein the Kaempferia pandurata extract is comprised in an amount of 0.1 to 10 weight % based on the total weight of the composition.

[Claim 13]

The composition for preventing or treating skin diseases according to claim 10, wherein the cosmetic composition is formulated in to skin adhesion type or transdermal type.

[Claim 14]

The composition for preventing or treating skin diseases according to claim 13, wherein the skin adhesion type formulation is softening toner, astringent toner, nourishing toner, nourishing cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, body lotion, body cream, body oil, body essence, makeup base, hair dye, shampoo, rinse or body cleaner.

[Claim 15]

The composition for preventing or treating skin diseases according to claim 13, wherein the transdermal type composition is lotion, ointment, gel, cream, patch or spray.