JP2012041276A - Carbonylation inhibitor of protein, or transparency improver of skin - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a protein carbonylation inhibitor or a skin transparency improver having a superior protein carbonylation inhibiting action and high safety.SOLUTION: Provided is a protein carbonylation inhibitor or a skin transparency improver which contains as an effective ingredient an extract from leaves of Engelhardtia chrysolepis Hance (juglandaceous evergreen tree) and/or an extract from lotus germ (nymphaesceous perennial aquatic plant ) originated in the tropics of Asia.

Description

  The present invention relates to a protein carbonylation inhibitor and a skin transparency enhancer containing components derived from natural products.

In recent years, researches on stratum corneum carbonylated proteins have been actively conducted in relation to aging and photoaging with skin aging. Generally, carbonylated proteins include those produced as a result of direct oxidation of NH ₂ groups of amino acid residues such as Lys, Arg and Pro in proteins to carbonyl groups, lipids oxidized to lipid peroxides, Is decomposed into a highly reactive aldehyde, which is produced by binding to protein (see Patent Document 1).

  In the skin, it is considered that protein oxidation may be initiated when protein is directly oxidized, or when sebum on the skin surface is oxidized by free radicals and lipid peroxide is generated. Once lipid peroxides are generated, oxidation proceeds in a chain, not only stimulating the skin surface, but also penetrates deep into the stratum corneum and damages cells.

  Further, the stratum corneum, which is the outermost layer of the skin, consists of keratinocytes, 85% of which is composed of keratin, a protein. In recent years, it has been known that this keratin is carbonylated by oxidative stress (ultraviolet rays, cigarette smoke) or the like that is routinely applied to the skin (see Non-Patent Document 1). Normally, keratin takes in a lot of water molecules, but carbonylation eliminates water molecules, and keratin after carbonylation cannot take up water molecules. It is known to dry the skin and damage the appearance of the skin (see Patent Document 2).

  It is known that the increase in carbonylated protein in the stratum corneum results in loss of skin transparency and water retention (see Patent Document 3) and loss of skin flexibility and elasticity. (See Patent Document 4).

  In this way, carbonylation of sebum and skin protein is thought to reduce all the functions inherent to the skin, which can maintain moisture, tension, brightness, etc. This is considered to be a cause of a decrease in skin transparency (see Non-Patent Document 2).

  Therefore, prevention of carbonylation of the horny layer protein of the epidermis or reduction of the degree of carbonylation is required for improving the skin transparency. That is, suppression of protein carbonylation is considered to be useful for improving water retention of keratinocytes and improving skin transparency.

  In addition, abnormal proteins such as carbonylated proteins increase with aging, and the accumulation of such abnormal proteins in vivo is Alzheimer's disease, Parkinson's disease, Lewy body disease, triplet repeat disease, amyotrophic lateral sclerosis. It is known to be involved in many diseases such as cataract, arteriosclerosis, diabetic nephropathy (Patent Document 5). Therefore, it is considered useful for preventing, treating or ameliorating diseases caused by carbonylated proteins by preventing / suppressing carbonylation of proteins.

  Thus, it is thought that the substance which can improve skin transparency and can suppress protein carbonylation is very useful. However, to date, no excellent substance that has such an action and has high safety and can be widely used as an ingredient for external preparations for skin, cosmetic foods and drinks, research reagents, etc. has not yet been provided. However, there is a strong demand for prompt provision.

JP 2004-340935 A JP 2004-107269 A JP-A-2005-249672 JP 2006-349372 A Japanese Patent No. 4255432

Jens J. Thiele et al., THE JOURNAL OF INVESTIGATIVE DERMATOLOGY, Vol.113, No.3, 1999, p.335, 339 Ichiro Iwai et al., J. Soc. Cosmet. Chem. Jpn., Vol.42, No.1, 2008, p.16-21

  An object of the present invention is to solve the above conventional problems and achieve the following objects. Specifically, an object of the present invention is to provide a protein carbonylation inhibitor having an excellent protein carbonylation inhibitory action and high safety. A second object of the present invention is to provide a skin transparency improving agent having an excellent skin transparency enhancing action and high safety.

  In order to solve the above-mentioned problems, the protein carbonylation inhibitor and skin transparency enhancer of the present invention are characterized in that they contain an extract from twilight leaves and / or an extract from lotus germ as active ingredients. To do.

  ADVANTAGE OF THE INVENTION According to this invention, it can provide the protein carbonylation inhibitor and skin transparency improvement agent which have the protein carbonylation suppression effect | action outstanding and high safety | security.

Hereinafter, embodiments of the present invention will be described in detail.
The protein carbonylation inhibitor or skin transparency enhancer of the present embodiment contains an extract from japonica leaves and / or an extract from lotus germ as an active ingredient.

  Here, in the present embodiment, the “extract” refers to an extract obtained using the above plant as an extraction raw material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or these Either a crude product or a purified product is included.

  In this embodiment, the extraction raw materials used for producing the extract from the yellow cocoon leaves and the extract from the lotus germ are jaundice (scientific name: Engelhardtia chrysolepis Hance) and lotus (scientific name: Nelumbo nucifera Gaertn.).

  Twilight (Engelhardtia chrysolepis Hance) is an evergreen tree plant belonging to the family Walnut. Twilight has been drunk as a kind of sweet tea since ancient times and is a highly safe plant. Twilight is native to regions from southern China to Taiwan and is readily available from these regions. Examples of the constituent parts of jaundice that can be used as an extraction raw material include branches, leaves, branches and leaves, and the like, preferably leaves.

  The lotus (Nelumbo nucifera Gaertn.) Is a perennial aquatic plant belonging to the genus Lotusaceae native to tropical Asia. In Japan, lotus root is eaten as a lotus root in Japan and is a highly safe plant. Lotus is distributed in southeastern Europe, northern Australia, eastern Asia, etc., and is cultivated in ponds, paddy fields, moats, etc., and can be easily obtained from these areas. The constituent part of the lotus used as an extraction raw material is germ. The lotus germ is a green, rod-shaped germ in a lotus seed.

  Although details of substances having an inhibitory action on protein carbonylation contained in extracts from twilight leaves and extracts from lotus germ are unknown, the above-mentioned plant is obtained by an extraction method generally used for plant extraction. From this, an extract having this action can be obtained.

  For example, after drying the plant, it is pulverized as it is or using a crusher, and is subjected to extraction with an extraction solvent, whereby an extract having a protein carbonylation-inhibiting action can be obtained. Drying may be performed in the sun or using a commonly used dryer. Moreover, after performing pretreatment, such as degreasing, with a nonpolar solvent such as hexane, it may be used as an extraction raw material. By performing pretreatment such as degreasing, extraction treatment with the polar solvent of the plant can be efficiently performed.

  As the extraction solvent, it is preferable to use a polar solvent, and examples thereof include water and hydrophilic organic solvents. These are used alone or in combination of two or more at room temperature or a temperature below the boiling point of the solvent. It is preferable.

  Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, and those subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, buffering, and the like. Therefore, the water that can be used as the extraction solvent in this embodiment includes purified water, hot water, ion exchange water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.

  Examples of hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene. Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycol, propylene glycol and glycerin.

  When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when using a mixed solution of water and a lower aliphatic alcohol, it is preferable to mix 1 to 90 parts by volume of a lower aliphatic alcohol with respect to 10 parts by volume of water. When using a mixed solution, it is preferable to mix 1 to 40 parts by volume of a lower aliphatic ketone with 10 parts by volume of water, and when using a mixed solution of water and a polyhydric alcohol, water 10 It is preferable to mix 10 to 90 parts by volume of a polyhydric alcohol with respect to the volume part.

  The extraction treatment is not particularly limited as long as the soluble component contained in the extraction raw material can be eluted in the extraction solvent, and can be performed according to a conventional method. For example, the extraction raw material is immersed in an extraction solvent 5 to 15 times (mass ratio) of the extraction raw material, the soluble components are extracted at room temperature or under reflux, and then filtered to remove the extraction residue. A liquid can be obtained. The obtained extract is diluted, concentrated, dried, purified, etc. according to a conventional method in order to obtain a diluted or concentrated solution of the extract, a dried product of the extract, or a crude purified product or a purified product thereof. Processing may be performed.

  Purification can be performed by, for example, activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, or the like. The obtained extract can be used as it is as an active ingredient of a protein carbonylation inhibitor or a skin transparency enhancer, but a concentrated solution or a dried product is easier to use.

  Since the extract obtained as described above has a unique odor and taste, it can be purified for the purpose of decolorization, deodorization, etc. within a range that does not cause a decrease in its physiological activity. However, since it is not used in a large amount when blended into an external preparation for skin, a cosmetic food or drink, etc., there is no practical problem even if it is not purified.

  The extract from yellow cocoon leaves and the extract from the lotus germ obtained as described above have an excellent protein carbonylation-inhibiting action. It can be used as an active ingredient. In addition, the extract from twilight leaves and the extract from the lotus germ can improve the transparency of the skin through the action of inhibiting the carbonylation of the protein, so it is also used as an active ingredient of the skin transparency enhancer. be able to.

  Furthermore, when a protein in the living body is carbonylated to produce a carbonylated protein and the carbonylated protein is accumulated in the living body, Alzheimer's disease, Parkinson's disease, Lewy body disease, triplet repeat disease, muscle atrophy Diseases such as lateral sclerosis, cataract, arteriosclerosis, diabetic nephropathy are thought to develop. Therefore, by utilizing the excellent protein carbonylation-inhibiting action of the extract from twilight leaves and the extract from lotus germ, the extract from twilight leaves and / or the extract from lotus germ is caused by protein carbonylation. It can be used as an active ingredient of a prophylactic / therapeutic agent for the above-mentioned diseases.

  In the present embodiment, any one of an extract from yellow cocoon leaves and an extract from lotus germ may be used as the active ingredient, or a mixture of both may be used as the active ingredient. . When an extract from twilight leaves and an extract from lotus germ are mixed and used as the active ingredient, the blending ratio may be appropriately adjusted depending on the degree of protein carbonylation-inhibiting action of these extracts.

  The protein carbonylation inhibitor or skin transparency enhancer of the present embodiment may be composed only of an extract from jaundice leaves and / or an extract from lotus germ, or an extract and / or lotus from jaundice leaves. It may be a formulation of an extract from embryo.

  The content of the extract from the yellow leaves and / or the extract from the lotus germ in the protein carbonylation inhibitor or skin transparency enhancer of the present embodiment is not particularly limited, and is appropriately determined depending on the purpose. Can be set.

  Extracts from twilight leaves and / or extracts from lotus germ are in the form of powder, granules, tablets in accordance with conventional methods using pharmaceutically acceptable carriers such as dextrin and cyclodextrin and other optional auxiliaries. It can be formulated into any dosage form such as liquid. In this case, as an auxiliary agent, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring / flavoring agent, and the like can be used. Extracts from twilight leaves and / or extracts from lotus germ can be used in combination with other compositions (for example, external preparations for skin, foods and drinks for cosmetics, etc.), ointments, liquids for external use, It can be used as a patch.

  In addition, the protein carbonylation inhibitor or the skin transparency enhancer of the present embodiment, if necessary, other extract or the like having a protein carbonylation inhibitory action, extract from yellow leaves and / or It can be used together with an extract from a lotus germ as an active ingredient.

  Examples of the administration method to the patient of the protein carbonylation inhibitor or skin transparency enhancer of the present embodiment include transdermal administration, oral administration, etc., depending on the type of the disease, for prevention and treatment thereof. A suitable method may be appropriately selected.

  The dose of the protein carbonylation inhibitor or skin transparency enhancer of the present embodiment may be appropriately increased or decreased depending on the type of disease, severity, individual differences among patients, administration method, administration period, and the like.

  The protein carbonylation inhibitor of the present embodiment can suppress carbonylation of stratum corneum proteins through the carbonylation-inhibiting action of the protein contained in the extract from japonica leaves and / or the extract from the lotus germ. Thereby, the skin beautification etc. can suppress the fall of the transparency of skin, and the skin-beautifying effect can be acquired. In addition, the protein carbonylation inhibitor of the present embodiment is a disease caused by protein carbonylation (Alzheimer's disease, through the protein carbonylation inhibitory action of an extract from japonica leaves and / or an extract from lotus germ. Parkinson's disease, Lewy body disease, triplet repeat disease, amyotrophic lateral sclerosis, cataract, arteriosclerosis, diabetic nephropathy, etc.) can be prevented and treated. However, the protein carbonylation inhibitor of the present embodiment can be used for all purposes that are meaningful for exerting a protein carbonylation-inhibiting action in addition to these uses.

  The skin transparency enhancer of the present embodiment can suppress the carbonylation of stratum corneum protein through the action of inhibiting the carbonylation of the protein possessed by the extract from twilight leaves and / or the extract from the lotus germ. Thus, the skin transparency can be improved. However, the skin transparency improving agent of the present embodiment can be used for all purposes other than these uses, which are meaningful for exerting a protein carbonylation-inhibiting action.

  The skin transparency enhancer and protein carbonylation inhibitor of the present embodiment are preferably applied to humans, but as long as the respective effects are exerted, animals other than humans are used. It can also be applied to (eg, mouse, rat, hamster, dog, cat, cow, pig, monkey, etc.).

  The protein carbonylation inhibitor and skin transparency enhancer of the present embodiment have an excellent protein carbonylation inhibitory action, and are excellent in usability and safety when applied to the skin. It is suitable for blending into an external preparation for skin.

  Here, the external skin preparation is not limited in its category, and includes a wide range of skin cosmetics, quasi-drugs, pharmaceuticals, etc. Specifically, for example, ointments, creams, milky lotions, cosmetic liquids, Lotions, packs, foundations, lip balms, bath salts, hair nicks, hair lotions, soaps, body shampoos and the like.

  In addition, the protein carbonylation inhibitor and skin transparency enhancer of the present embodiment have an excellent protein carbonylation inhibitory action and are excellent in safety when taken orally, for example, It is suitable for blending into beauty foods and beverages. Here, there is no restriction | limiting in the classification | category as food / beverage products for beauty, and it includes a wide range of foods such as general foods, health foods, health functional foods, quasi drugs, and pharmaceuticals that are taken orally.

  Furthermore, since the protein carbonylation inhibitor and skin transparency enhancer of the present embodiment have excellent protein carbonylation inhibitory action, they are also suitable as reagents for the study of diseases related to protein carbonylation. Can be used.

  Hereinafter, although a test example is given and this invention is demonstrated further in detail, this invention is not limited to the following test example at all. In addition, in this test example, an extract from yellow cocoon leaf was used as an extract from yellow cocoon leaf (manufactured by Maruzen Pharmaceutical Co., Ltd., sample 1), and an extract from lotus germ was used as a lotus germ extract powder (manufactured by Maruzen Pharmaceutical Co., Ltd., sample 2). used.

[Test Example 1] Test for inhibiting carbonylation of stratum corneum protein With respect to the extract from the above yellow leaves and the extract from lotus germ (sample 1 and sample 2), the carbonylation inhibitory action of the stratum corneum protein was obtained by the following method. Tested.

  The cheek layer adhesive tape (product name: horny checker, manufactured by Asahi Biomed) was pressed against the inner side of the upper arm, and the horny layer (keratinocytes) was collected. A sample solution (sample) in which 0.02 mL of a solution obtained by dissolving the above samples (sample 1 and sample 2) in a 50 vol% ethanol aqueous solution was added to 0.98 mL of a 20 μmol / L sodium hypochlorite (Wako Pure Chemical Industries, Ltd.) aqueous solution. The concentration tape was immersed in the adhesive tape at 37 ° C. for 16 hours to perform carbonylation treatment of the stratum corneum protein, and the carbonylation level was evaluated by the following method.

  Apply the above adhesive tape to fluorescent hydrazide (fluorescein-5-thiosemicarbazide, manufactured by AnaSpec) adjusted to a concentration of 20 μmol / L with 0.1 mol / L 2-morpholinoethanesulfonic acid-Na (pH 5.5) buffer solution. It was immersed, and the carbonyl group of the stratum corneum protein carbonylated by carbonylation treatment was labeled.

  The above adhesive tape is imaged (imaging device: fluorescence microscope Olympus IX71, manufactured by Olympus), and the obtained observation image is analyzed (analysis software: ImageJ, manufactured by National Institute of Health). Was the carbonylation level. In addition, as a control, the same test was performed on the carbonylation treatment of the stratum corneum protein by immersing the adhesive tape in an aqueous solution of 20 μmol / L sodium hypochlorite (manufactured by Wako Pure Chemical Industries, Ltd.) without adding a sample. .

From the obtained results, the carbonylation inhibition rate (%) of the stratum corneum protein was calculated by the following formula.
Carbonylation inhibition rate (%) = {(A−B) / (A−C)} × 100
In the formula, A represents “fluorescence luminance when sodium hypochlorite is added and no sample is added”, B represents “fluorescence luminance when sodium hypochlorite is added and sample is added”, and C is “hypochlorite” “Fluorescence luminance when no sodium acid is added and no sample is added”.
The results are shown in Table 1.

  As shown in Table 1, it was found that the extract from the japonica leaves and the extract from the lotus germ (sample 1 and sample 2) have a carbonylation-inhibiting action on stratum corneum proteins. It was suggested that the extracts from the lotus germ (Sample 1 and Sample 2) have an effect of improving skin transparency.

  The protein carbonylation inhibitor and skin transparency improver of the present invention have an excellent protein carbonylation inhibitory action and are excellent in safety. It is useful as a research reagent.

Claims (2)

  A protein carbonylation inhibitor comprising an extract from twilight leaves and / or an extract from lotus germ as an active ingredient.   An agent for improving skin transparency, comprising an extract from twilight leaves and / or an extract from lotus germ as an active ingredient.