JP2011162507A - Inhibitor of extension/stimulation-mediated production of collagen decomposition enzyme - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an inhibitor of extension/stimulation-mediated production of a collagen decomposition enzyme that has effects of inhibiting extension/stimulation-mediated production of a collagen decomposition enzyme and preventing wrinkles, and is used as external preparations for skin, cosmetics and quasi drugs excellent in safety and feeling of use. <P>SOLUTION: The inhibitor of extension/stimulation-mediated production of a collagen decomposition enzyme contains an extract of a plant of Avena fatua genus or an extract of a plant of Psoralea corylifolia genus. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention controls the production of collagen-degrading enzymes mediated by extension stimulation, which can suppress the formation of wrinkles in areas where skin contraction is likely to occur, such as the corners of the eyes, forehead, and mouth, by controlling the production of collagen-degrading enzymes that are generated by stretching stimulation. The present invention relates to an external preparation for skin, a skin cosmetic, and a quasi-drug containing an agent and an agent for inhibiting the production of collagen degradation enzyme mediated by extension stimulation.

  Pigmental abnormalities in the skin include hyperpigmentation such as spots (liver spots), freckles (sparrow egg spots), senile pigment spots; depigmentation such as vulgaris, depigmented nevus, and albinism Are known.

  It is known that such a pigment abnormality is caused by, for example, deposition of melanin, which is a pigment generated in melanocytes existing in the epidermis basal layer of the skin. That is, melanocytes in the skin are present in the basal layer of the lowermost layer of the epidermis and play a role of protecting the skin from solar ultraviolet rays. However, excessive melanin production results in pigmentation, causing spots (liver spots), freckle (spotted egg spots), and senile pigment spots, which is one of the skin problems. The mechanism of action of these pigmentations is generally triggered by stimulation of ultraviolet rays from sunlight, abnormal hormone balance, inflammation, etc., which activates melanin production in melanocytes, It is thought to deposit abnormally.

  The treatment in the mechanism of action of pigmentation is roughly classified into melanin pigment production suppression, melanin pigment reduction, and melanin pigment excretion promotion. Based on this idea, various patent applications have been filed. For example, Patent Document 1 discloses an invention relating to an external preparation found by finding that arbutin exhibits a melanin production inhibitory effect, and Patent Document 2 finds that glabrizine exhibits a tyrosinase activity and is a melanin production inhibitor. An invention relating to an external preparation is disclosed. In addition, as a reducing agent for melanin pigment, for example, a patent application using ascorbic acid as in Patent Document 3 is made, and a patent application like Patent Document 4 using glutathione has been made.

  However, arbutin, glabrizine, ascorbic acid, glutathione, etc. may not be able to sufficiently exert the inhibitory effect on melanin production, and have not been obtained to the extent that the effect on the action mechanism of pigmentation is expected.

  By the way, in recent years, skin extension stimulation has attracted attention as a new mechanism of pigmentation. Stretch stimulation increases the amount of GM-CSF produced from epidermal keratinocytes by applying an external mechanical stimulus that stretches the skin and activates melanocytes, thereby causing excessive melanin synthesis. It is what causes. Therefore, it is considered that pigmentation can be improved more effectively if a component that inhibits production of GM-CSF enhanced by extension stimulation can be found.

  The present inventors have paid attention to such a skin extension stimulus, filed a patent application such as Patent Document 5 relating to a method for evaluating a substance that controls extension stimulus-mediated melanin production, and further promoted by extension stimulus GM- A component that inhibits CSF production was found, and a patent application such as Patent Document 6 was filed.

  The inventors hypothesized that collagen degradation enzymes may be involved as a mechanism of facial wrinkles due to skin stretch stimulation from a viewpoint different from the suppression of melanin production as described above. Below, as a result of intensive studies to suppress the production of collagen degrading enzymes due to such skin stretching stimulation, the matrix metalloprotease-1 (hereinafter referred to as MMP-1), which is a collagen degrading enzyme, is reduced. Such an active ingredient has been found and the present invention has been completed.

JP 63-8314 A Japanese Unexamined Patent Publication No. 01-311011 JP 2003-104864 A Japanese Patent Laid-Open No. 11-269058 JP 2008-203324 A JP 2009-108115 A

  The present invention suppresses the production of collagenolytic enzymes due to stretching stimulation, has the effect of preventing the generation of wrinkles, and is used as a skin external preparation, cosmetic, and quasi-drug excellent in safety and usability. It is an object of the present invention to provide a stimulus-mediated collagenase production inhibitor.

In order to solve such a problem, the present invention provides an agent for suppressing the production of collagen degradation enzyme mediated by extension stimulation, characterized by containing an extract of a crow genus plant or an extract of a Dutch genus plant. To do. Moreover, this invention provides the skin external preparation, cosmetics, and quasi-drug which mix | blended such an extension stimulation mediated collagenase production inhibitor. For example, oats ( Avena sativa ) and oats ( Avena fatua ) are used as the genus Crows, and for example, Psoralea corylifolia is used as the Dutch genus plant.

  According to the present invention, by reducing MMP-1, which is a collagen degrading enzyme in the dermis, collagen degradation is suppressed, thereby preventing and improving the generation of wrinkles, and highly safe extension. It is possible to provide an external preparation for skin, a cosmetic, and the like containing a stimulus-mediated collagenase degrading enzyme inhibitor, and a stretching stimulus-mediated collagenase degrading enzyme production inhibitor.

Hereinafter, embodiments of the present invention will be described.
As described above, the stretch stimulation-mediated collagenase production inhibitor of the present invention contains an extract of the genus Oat plant or an extract of the genus Orna plant. Here, “containing” means that the agent for inhibiting the production of collagen degradation enzyme mediated by stretch stimulation of the present invention is composed of an extract of a genus oat plant, or an extract of an genus dutch plant, in addition to these. It means that things other than the extract may be contained.

The “crown plant”, which is one of the stretch stimulation mediated collagenase production inhibitors of the present invention, is a plant belonging to the grass family. Examples of crow oats include oat ( Avena sativa ), oat ( Avena fatua ), hadaka oat ( Avena nuda ), Chahi ( Avena strigosa ), Avena byzantina , Avena abyssinica , Avena barbata , Avena brevis, etc. However, in the present invention, it is particularly desirable to use oat ( Avena sativa ). Although it does not specifically limit as an extraction site | part in the case of extracting an extract from a crow oat plant, It is preferable to use a grain.

In addition, “Netherlands plant” which is another one of the stretch stimulation mediated collagenase production inhibitors of the present invention is a leguminous plant. An example of the “Netherland genus plant” is a Dutch villa ( Psoralea corylifolia ). The extraction site in the case of extracting an extract from a plant of the genus Calla is not particularly limited, but it is preferable to use a fruit or a seed.

  The various parts of each plant body in the plant extract used in the present invention are preferably the parts described above, but in addition to this, selected from flowers, flower ears, fruit skin, fruits, stems, leaves, root bark, roots, whole plants, etc. 1 type (s) or 2 or more types can be used. Moreover, you may utilize what can be obtained as a crude drug. In addition to those obtained by direct extraction from these various extraction sites using a solvent, the extract is obtained by adding a solvent to the squeezed liquid and / or residue obtained after the squeezing treatment. Are also included in the scope of the extract in the present invention.

  As a solvent used for extraction, any solvent can be used as long as it is a solvent used for normal plant extraction. For example, water, alcohols such as methanol, ethanol, propylene glycol, 1,3-butylene glycol, glycerin, hydrous alcohols, organic solvents such as chloroform, dichloroethane, carbon tetrachloride, acetone, ethyl acetate, hexane, etc. They can be used alone or in combination.

  The compounding amount of the extract of the genus Oats plant or the plant of the Dutch genus plant is usually preferably 0.000001 to 5% by weight as a dry solid content. If the amount is less than 0.000001% by weight, the effect of the present invention may not be sufficiently obtained. On the other hand, if the amount exceeds 5% by weight, an improvement in the effect commensurate with the increase is not recognized. From this viewpoint, 0.00001 to 1% by weight is more preferable.

  In the stretch stimulation-mediated collagenase production inhibitor of the present invention, various optional components that are generally used in cosmetics or used in skin external preparations such as quasi-drugs and pharmaceuticals, as long as the effects of the present invention are not impaired. Can be appropriately blended as necessary. Examples of such optional ingredients include purified water, ethanol, oily ingredients, moisturizers, thickeners, preservatives, emulsifiers, medicinal ingredients, powders, ultraviolet absorbers, dyes, fragrances, and emulsion stabilizers. Can do.

  The form of the stretch stimulation-mediated collagenase production inhibitor in the present invention is not limited as long as it is applicable to the outer skin such as liquid, emulsion, ointment, cream, gel, aerosol, and the like as appropriate. A base component etc. can be mix | blended and a desired form can be prepared. In addition, the stretch stimulation-mediated collagenolytic enzyme production inhibitor of the present invention is applicable in various fields such as pharmaceuticals, quasi drugs, and cosmetics.

  In the present invention, “extension stimulation-mediated collagenolytic enzyme production” refers to the production of collagenolytic enzyme resulting from the load of the extension stimulus. The “extension stimulation-mediated collagenolytic enzyme production inhibitor” of the present invention is a collagenolytic enzyme production inhibitor that inhibits the production of collagenolytic enzymes due to the load of the extension stimulus as described above. The stretch stimulation mediated collagenase production inhibitor of the present invention can inhibit the production of collagenase that causes wrinkles and the like in areas where skin contraction is likely to occur, such as the corners of the eyes, the forehead, and the mouth.

  Such contraction of the skin of the corners of the eyes, the forehead, the mouth, etc. occurs naturally, predicts when the load of the extension stimulus due to such contraction of the skin occurs, or whether the load of the extension stimulus occurs It is not easy to confirm such. Therefore, it is not easy to confirm the inhibitory effect on the production of collagenolytic enzyme caused by such a naturally occurring extension stimulus load with respect to the agent for inhibiting the production of collagenolytic enzyme mediated by the extension stimulus of the present invention.

  However, it is possible to artificially generate and check such a load of extension stimulation. As means for artificially generating such a load of extension stimulation, means capable of extending cells at a predetermined interval, for example, means such as a cultured cell extension apparatus as described in the following examples can be employed. Such a stretching stimulus is preferably a stimulus that periodically stretches cells in a uniaxial direction from the viewpoint of the directionality and frequency of stretching in the skin.

  The degree of such extension is 1.01 to 1.40 times the size of the culture container under normal culture conditions (conditions in which no extension stimulus is loaded), preferably from the viewpoint of the stretch ratio in the skin. The condition of 1.04 to 1.20 times is desirable. When such extension stimulation is performed using ST-140 (trade name: manufactured by Strex Co., Ltd.), which is one of the cultured cell extension apparatuses as in the examples described later, for example, the conditions for the extension stimulation are as follows. Examples include a condition in which 10% extension stimulation is performed for 8 days twice in one minute and in a uniaxial direction.

  As cultured cells in this case, for example, NHDF (human fibroblast) can be used. Moreover, as a culture medium, the culture medium suitable for culture | cultivation of the cell used is mentioned. Examples thereof include Dulbecco's modified Eagle medium (DMEM) containing fetal calf serum, Eagle medium containing fetal calf serum, Medium 153 medium containing various growth factors such as fibroblast growth factor (FGF), and the like. In particular, when a cell line is used, it is desirable to use Dulbecco's modified Eagle medium containing fetal calf serum or an Eagle medium containing fetal calf serum.

  By culturing such cells, it is possible to confirm the correlation between skin stretch stimulation and the production of collagenolytic enzymes.

  The amount of MMP-1 can be measured, for example, by recovering the culture supernatant after culturing, and quantifying MMP-1 contained in the culture supernatant by ELISA using a monoclonal antibody against MMP-1. .

  In addition, the cells on the 8th day after the skin stretching stimulation load were collected, and mRNA in the cells was collected, cDNA complementary to the mRNA was synthesized, and the cDNA was quantified by a real-time RT-PCR method. As a result, the amount of MMP-1 can also be measured by quantifying mRNA.

  Examples of the present invention will be described below.

Example 1
In this example, an extract of oat ( Avena sativa ) was used as an example of an agent for suppressing production of collagen degradation enzyme mediated by stretch stimulation. The preparation method will be described. First, 100 g of dried oat kernel pulverized product was immersed in a 10-fold amount of 50 vol% 1,3-butylene glycol solution and left at room temperature for 5 days and nights. After the extraction, filtration was performed to obtain an oat extract (dry solid content: about 0.6% by mass).

(Example 2)
In this example, an extract of Dutch bay ( Psoralea corylifolia ) is used as an example of an agent for suppressing the production of collagen degradation enzyme mediated by extension stimulation. The preparation method was the same as in Example 1. 100 g of dried Dutch ground seeds were dipped in 10 volumes of 50% by volume ethanol solution, extracted at room temperature for 5 days and nights, and then filtered to obtain Dutch belly extract. (Dry solid content: about 0.5 mass%) was obtained.

(Test Example 1)
The production test of MMP-1 using NHDF (human fibroblast) was performed on the stretch stimulation-mediated collagenase production inhibitor of Examples 1 and 2 prepared as described above. Quantification of MMP-1 was performed by ELISA. The test method is as follows.

That is, first, in a type 1 collagen-coated silicon chamber, NHDF was seeded and 3 ml of Dulbecco's modified Eagle medium (hereinafter referred to as “fetal bovine serum-containing DMEM”) containing 10% by mass fetal bovine serum was used as the medium. NHDF was cultured for 24 hours under the conditions of 5% by volume CO ₂ and 37 ° C. Next, the culture medium in the silicon chamber was removed, and then the silicon chamber was set in a cultured cell extension apparatus [trade name: ST-140, manufactured by Strex Corporation].

  The oat extract obtained in Example 1 above was newly added to DMEM containing 10% by mass fetal bovine serum so as to obtain a final concentration of 0.5% to obtain a medium. Similarly, the Dutch sweetfish extract obtained in Example 2 was added to a DMEM containing 10% by weight fetal calf serum so that the final concentration was 1% to obtain a new medium. On the other hand, a DMEM containing 10% by mass fetal bovine serum was prepared as a comparative example without adding a stretch stimulation-mediated collagenase production inhibitor.

  In this way, the medium added with the growth stimulation-mediated collagenolytic enzyme production inhibitor of Examples 1 and 2 and the medium prepared as a comparative example were added to the silicon chamber set in the cultured cell extension apparatus, and added for 1 minute. Twice a uniaxial direction was applied with 10% extension stimulation for 8 days. Here, the “10% extension stimulation” means that the culture area becomes 1.1 times by extending the silicon chamber to the left and right.

  On the 3rd and 7th day after the extension stimulation load, the oat extract obtained in Example 1 is newly added to DMEM containing 10% by mass fetal calf serum so that the final concentration becomes 0.5%. The medium was replaced with the prepared medium. In addition, the Dutch sweetfish extract obtained in Example 2 was added to the medium added to DMEM containing 10% by mass fetal bovine serum so that the final concentration was 1% on the third and seventh days after the extension stimulation. The medium was changed. Thereafter, the culture supernatant was obtained on the 8th day, and the cells were detached from the silicon chamber and collected using a mixed solution of trypsin and EDTA.

  The collected cells were suspended in 1 ml of PBS, and the cells were counted with a hemocytometer. The collected supernatant was quantified by ELISA method for the amount of MMP-1 produced in the culture supernatant. The measurement was performed using an hMMP-1 measurement kit “Daiichi Fine Chemical Co., Ltd.” according to the method described in the attached manual. The MMP-1 production rate was calculated with the amount of MMP-1 in the case of the comparative example cultured in a medium not added with the stretch stimulation-mediated collagenolytic enzyme production inhibitor under the stretch stimulus load as 100. The test results are shown in Table 1.

  As is clear from Table 1, the addition of 0.5% by mass and 1% by mass of the oat extract (Example 1) and the Dutch biloba extract (Example 2), respectively, resulted in MMP by NHDF under an extension stimulus load. The production rate of -1 was less than that of the comparative example. From this, it was found that the collagenase production inhibitor mediated by stretch stimulation in Examples 1 and 2 has a production inhibitory effect on MMP-1.

  Next, formulation examples of the cosmetic composition of the present invention, in which the extension stimulation-mediated collagenolytic enzyme production inhibitor of Examples 1 and 2 is blended, are shown below.

(Formulation example 1) Skin care lotion Composition Composition ratio (mass%)
Ethanol 5.0%
1,3-butylene glycol 7.0%
Sorbit solution 10.0%
Oat extract of Example 1 0.01%
Methyl paraoxybenzoate 0.15%
Citric acid 0.05%
Sodium citrate 0.2%
Edetate 0.05%
Fragrance 0.1%
POE hydrogenated castor oil 0.5%
Purified water remaining

(Formulation example 2) Skin care lotion Composition ratio (% by mass)
Ethanol 5.0%
1,3-butylene glycol 7.0%
Sorbit solution 10.0%
0.01% Dutch bay extract from Example 2
Methyl paraoxybenzoate 0.15%
Citric acid 0.05%
Sodium citrate 0.2%
Edetate 0.05%
Fragrance 0.1%
POE hydrogenated castor oil 0.5%
Purified water remaining

(Formulation example 3) Skin care cream Composition Formulation ratio (mass%)
Cetanol 2.5%
Squalane 10.0%
White beeswax 1.0%
Glyceryl trioctanoate 5.0%
Octyldodecyl myristate 15.0%
Tocopherol acetate 0.1%
1,3-butylene glycol 7.0%
Glycerol monostearate 3.0%
POE (20) sorbitan monostearate 1.0%
Sorbitan monostearate 2.0%
Oat extract of Example 1 0.1%
Concentrated glycerin 5.0%
Butyl paraoxybenzoate 0.1%
Methyl paraoxybenzoate 0.2%
Purified water remaining

(Formulation example 4) Skin care cream Composition Composition ratio (% by weight)
Cetanol 2.5%
Squalane 10.0%
White beeswax 1.0%
Glyceryl trioctanoate 5.0%
Octyldodecyl myristate 15.0%
Tocopherol acetate 0.1%
1,3-butylene glycol 7.0%
Glycerol monostearate 3.0%
POE (20) sorbitan monostearate 1.0%
Sorbitan monostearate 2.0%
0.1% Dutch bay extract of Example 2
Concentrated glycerin 5.0%
Butyl paraoxybenzoate 0.1%
Methyl paraoxybenzoate 0.2%
Purified water remaining

(Other examples)
In addition, the preparation method of the collagen-stimulating enzyme production inhibitor mediated by extension stimulation in each of the above Examples 1 and 2 is not limited to the method described in Examples 1 and 2, and is prepared by other methods. Is also possible.

  Further, in Examples 1 and 2, the case where the collagen-degrading enzyme production inhibitor mediated by extension stimulation is composed of only an oat extract and a Dutch beetle extract, respectively, but these two or more types are included to cause extension stimulation. It is also possible to use an intervening collagenolytic enzyme production inhibitor. Moreover, it is also possible to contain things other than these oat extracts and Dutch beetle extracts in a collagen-degrading enzyme production inhibitor mediated by stretch stimulation.

  Furthermore, in the above-mentioned prescription examples 1 to 4, the case where a collagen-degrading enzyme production inhibitor mediated by extension stimulation is added to skin care lotions and skin care creams has been described. However, collagen other than skin care lotions and cosmetics other than skin care creams. It is also possible to mix a degradation enzyme production inhibitor. In addition, it is possible to blend the agent for inhibiting the production of collagen degradation enzyme mediated by the stretch stimulation of the present invention into an external preparation for skin other than cosmetics, and it can also be blended into a quasi drug.

Claims (6)

  An agent for suppressing production of collagen degradation enzyme mediated by extension stimulation, comprising an extract of a crow barley plant or an extract of a Dutch genus plant. The growth stimulating-mediated collagenolytic enzyme production inhibitor according to claim 1, wherein the crow barley plant is oat ( Avena sativa ) or oat ( Avena fatua ). 3. The stretch stimulation-mediated collagenolytic enzyme production inhibitor according to claim 1 or 2, wherein the Dutch genus plant is Dutch biloba ( Psoralea corylifolia ).   An external preparation for skin comprising the stretch stimulation-mediated collagenase production inhibitor according to any one of claims 1 to 3.   A cosmetic comprising the stretch stimulation-mediated collagenase production inhibitor according to any one of claims 1 to 3.   A quasi-drug containing the stretch stimulation-mediated collagenase production inhibitor according to any one of claims 1 to 3.