JP2017206468A - Lysosome activity promotor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a novel lysosome activity promotor.SOLUTION: A lysosome activity promotor is composed of extract from plants belonging to Poaceae, Coix and/or extract from plants belonging to Betulaceae, Betula.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a lysosome activity promoter.

  The lysosome is one of the organelles of eukaryotes and is a place for intracellular digestion with hydrolase inside. Biopolymers that are taken into the membrane by endocytosis and autophagy are degraded in lysosomes. Useful degradation products are absorbed and reused in the cytoplasm, and waste products are discarded outside the cell by exocytosis. Or stay in the cell as a residual body.

  It is known that lysosomal activity is involved in various diseases. As a typical disease, lysosomal storage disease is known. In many cases, lysosomal storage diseases are caused by accumulation of a substrate of the enzyme in a cell as a deposit without being degraded due to a deficiency or mutation of a hydrolase acting in the lysosome. Lysosomal storage diseases have a wide range of clinical heterogeneity and biological diversity, and include lipid storage diseases, mucopolysaccharidosis, mucolipidosis, and glycoprotein storage diseases (see Patent Documents 1 and 2). .

  On the other hand, it is known that a whitening effect can be obtained by improving lysosomal activity in keratinocytes and promoting degradation of melanosome proteins (Patent Document 3, Non-Patent Documents 1 and 2).

  In addition, cathepsins B1 and D, which are lysosomal enzymes, degrade collagen (Non-patent Document 3), hyaluronic acid is taken into lysosomes and degraded (Non-patent Document 4), compared to younger skin It is known that the amount of fragmented collagen is significantly higher in the skin of elderly people (Non-patent Document 5).

By the way, hyaluronic acid is a mucopolysaccharide which is present abundantly in the dermis of the skin, and contributes to moisture retention and elasticity of the skin. And it is known that the amount will decrease with aging, and the material which accelerates | stimulates the production | generation of hyaluronic acid has been utilized for the cosmetics etc. for the purpose of anti-aging and moisturizing conventionally.
For example, pollen or pollen extract of cistus, tea, rapeseed, and buckwheat is known to have a hyaluronic acid production promoting effect and is known to be usable as an anti-aging skin external preparation (Patent Document 4). .

Special table 2014-517015 gazette Special table 2014-523881 gazette Japanese Patent Laid-Open No. 2015-10071 JP 2008-133270 A

FUJIFILM Corporation, “demonstrating the phenomenon that melanin pigment causing stains is decomposed in epidermal cells. Confirmation of melanin degradation promoting effect on whitening useful ingredient“ AMA ””, [online], November 2013 May 28, [Search March 24, 2016], Internet <URL: http://www.fujifilm.co.jp/corporate/news/articleffnr_0831.html> FUJIFILM Corporation, “Proteolytic enzyme“ cathepsin V ”found to be involved in the degradation of melanosomes, including melanin pigments that cause stains,” Orizanol, contained in rice bran lipids, promotes the degradation of melanosomes "Confirmation", [online], November 5, 2015, [March 24, 2016 search], Internet <URL: http://www.fujifilm.co.jp/corporate/news/articleffnr_1020.html> Biochem. J. (1974) 137, 387-398 Matrix Biol. 2002 Jan; 21 (1): 15-23. The Journal of Investigative Dermatology vol. 120, No. 5 may 2003.

  As described above, lysosome is related to various diseases, and development of a component capable of improving its activity is desired. Then, this invention makes it a 1st subject to provide a novel lysosome activity promoter.

  Further, as described above, hyaluronic acid is known to decrease with aging, and development of a component that promotes the production of hyaluronic acid is desired. Then, this invention makes it a 2nd subject to provide a novel hyaluronic acid production promoter.

  It is a third object to provide a novel screening method for components having hyaluronic acid production promoting action.

As a result of diligent research efforts, the present inventors have developed extracts of plants belonging to the genus Poaceae (Coix), extracts of plants belonging to the family Betulaceae (Betula), and mixtures thereof. It was found that there is an action to improve the activity of lysosome.
Based on the above findings, the present inventors have completed the present invention.

The present invention for solving the first problem is an lysosome comprising an extract of a plant belonging to the genus Poaceae (Coix) and / or an extract of a plant belonging to the genus Betulaceae (Betula). It is an activity promoter.
The lysosomal activity promoter of the present invention is useful for the treatment or prevention of diseases caused by a decrease in lysosomal activity and the improvement of biological functions related to lysosomal activity.

In a preferred form of the enzyme activator of the present invention, the enzyme activator comprises a mixture of an extract of a plant belonging to the genus Poaceae (Coix) and an extract of a plant belonging to the genus Betulaceae (Betula).
According to this form of the present invention, a more excellent lysosomal activity promoting effect can be obtained by the synergistic effect of the two plant extracts.

  In a preferred embodiment of the present invention, the plant belonging to the family Poaceae (Coix) is barley (Coix lacryma-jobi var. Ma-yuen), and the plant belonging to the family Betulaceae (Betula). Is birch (Betula platyphylla).

  The lysosomal activity promoter of the present invention is excellent in hyaluronic acid production promoting action. Therefore, the lysosomal activity promoter of the present invention can be used for promoting hyaluronic acid production.

  The lysosome activity promoter of the present invention is preferably in the form of an external preparation or an oral preparation.

Further, as a result of earnest research efforts, the present inventors have obtained a mixture of an extract of a plant belonging to the genus Poaceae (Coix) and an extract of a plant belonging to the genus Betulaceae (Betula), It has been found that it has an effect of promoting hyaluronic acid production.
Based on the above findings, the present inventors have completed the present invention.

The present invention that solves the second problem is an extract of a plant belonging to the genus Poaceae (Coix) and / or an extract of a plant belonging to the genus Betulaceae (Betula). It is an acid production promoter.
The preferred form of the hyaluronic acid production promoter of the present invention is the same as that of the invention relating to the lysosome activity promoter described above.

The present invention for solving the third problem is a screening method for a component having hyaluronic acid production promoting action, characterized by using lysosomal activity as an index.
According to the present invention, a component having a hyaluronic acid production promoting action can be screened by measuring lysosomal activity without directly measuring the amount of hyaluronic acid produced.

The lysosome activity promoter of the present invention is excellent in the action of improving lysosome activity.
Moreover, the hyaluronic acid production promoter of the present invention is excellent in the effect of promoting hyaluronic acid production.

10 is a bar graph showing the results of Test Example 1. It represents lysosomal activity when a pearl barley extract, birch extract or a mixture thereof is added to a fibroblast culture. 10 is a bar graph showing the results of Test Example 2. It represents lysosomal activity when a mixture of pearl barley extract and birch extract is added to the epidermal cell culture. 10 is a bar graph showing the results of Test Example 2. The figure shows lysosomal activity when a mixture of pearl barley extract and birch extract is added to the culture solution of fibroblasts. 10 is a bar graph showing the results of Test Example 3. It represents the gene expression level of hyaluronic acid synthase after adding pearl barley extract, birch extract or a mixture thereof to the culture solution of epidermal cells and culturing overnight. As a comparative example, gentian, a mixture of gentian and pearl barley, and a mixture of gentian and birch were added to the culture solution of epidermal cells. 10 is a bar graph showing the results of Test Example 3. It represents the gene expression level of hyaluronic acid synthase after overnight addition of pearl barley extract, birch extract or a mixture thereof to the fibroblast culture. 10 is a bar graph showing the results of Test Example 4. It shows the amount of hyaluronic acid produced after culturing with the mixture of Leupeptin and Pepstatin, which are lysosomal activity inhibitors, in the culture solution of epidermal cells. 10 is a bar graph showing the results of Test Example 4. The amount of hyaluronic acid produced after adding a mixture of Leupeptin and Pepstatin, which are lysosomal activity inhibitors, to a culture of fibroblasts and culturing.

<1> Lysosome Activity Promoter The lysosome activity promoter of the present invention is an extract of a plant belonging to the Poaceae family (Coix) and / or a plant belonging to the Betulaceae family (Betula). It consists of things.

Examples of the plant belonging to the genus Poaceae (Poixae) used in the present invention include Coix lachryma-jobi L. and its variants, Coix lachryma-jobi var. Maxima Makino, Naga Examples include Coix lachryma-jobi var.stenocarpa Stapf. And pearl barley (Coix lachryma-jobi var.ma-yuen). Is particularly preferred.
The extraction site for obtaining an extract of a plant belonging to the genus Juzudama is not particularly limited, and the extract is obtained using one or more selected from whole grass or leaves, stems, roots, flowers, and seeds. be able to. Among these, an extract obtained from seeds is particularly preferable.

Examples of plants belonging to the genus Betulaceae (Betula) used in the present invention include birch (Betula platyphylla), North American birch (Betula lenta), European birch (Betula pendula) and the like. Betula platyphylla) is particularly preferred.
The extraction site for obtaining an extract of a plant belonging to the genus Birch is not particularly limited, and the extract can be obtained using one or more selected from leaves, bark, and xylem of plants. Among these, an extract obtained from bark is particularly preferable.

  The plant extract in the present invention can be prepared by using a plant grown in Japan, a plant grown in Japan, or a product from Japan sold as a herbal medicine raw material, or a plant such as Maruzen Co., Ltd. It is also possible to purchase and use a commercial extract sold by a company that handles the extract.

  In the extraction, the plant body, the above-ground part, the tree trunk part, the rhizome part or the seed is preferably processed in advance so as to improve the extraction efficiency by crushing or chopping. For the extract, 1 to 30 parts by mass of a solvent is added to 1 part by mass of the plant body, the above-ground part, the tree trunk part, the rhizome part or the seed or the dried product thereof. Immerse for several hours. After the immersion, the solution can be cooled to room temperature, insolubles can be removed if desired, and then the solvent can be removed by concentration under reduced pressure. Thereafter, fractionation and purification can be performed by column chromatography packed with silica gel or ion exchange resin to obtain a desired extract.

  The extraction solvent is preferably a polar solvent, and alcohols such as water, ethanol, isopropyl alcohol, and butanol, polyhydric alcohols such as 1,3-butanediol, and polypropylene glycol. One or two or more selected from ketones such as acetone, methyl ethyl ketone, and ethers such as diethyl ether and tetrahydrofuran can be preferably exemplified.

It is preferable that the lysosome activity promoter of the present invention includes a mixture of an extract of a plant belonging to the genus Grazaceae and an extract of a plant belonging to the genus Birchaceae. By setting it as such embodiment, the more superior lysosome activity promotion effect | action can be obtained by the synergistic effect of the extract of two types of plants.
In this case, the dry mass ratio of the extract of the plant belonging to the genus Grazaceae and the plant belonging to Birchaceae in the lysosome activity promoter is preferably 1000: 1 to 1: 1000, more preferably 100: 1 to 1. : 100, more preferably 20: 1 to 1:20, more preferably 10: 1 to 1:10.

The lysosome activity promoter of the present invention is preferably in the form of an external preparation or an oral preparation.
As external preparations, cosmetics, quasi-drugs, pharmaceuticals and the like can be suitably exemplified, and any commonly used component can be contained as long as the effects of the present invention are not impaired. Examples of such optional ingredients include macadamia nut oil, avocado oil, corn oil, olive oil, rapeseed oil, sesame oil, castor oil, safflower oil, cottonseed oil, jojoba oil, coconut oil, palm oil, liquid lanolin, and hardened coconut oil. Oil, wax, oils such as beeswax, owl, hydrogenated castor oil, beeswax, candelilla wax, carnauba wax, ibotarou, lanolin, reduced lanolin, hard lanolin, jojoba wax; liquid paraffin, squalane, pristane, ozokerite, paraffin, ceresin, petrolatum , Hydrocarbons such as microcrystalline wax; higher fatty acids such as oleic acid, isostearic acid, lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, undecylenic acid; cetyl alcohol, stearyl alcohol, isostear Higher alcohols such as alcohol, behenyl alcohol, octyldodecanol, myristyl alcohol, cetostearyl alcohol; cetyl isooctanoate, isopropyl myristate, hexyldecyl isostearate, diisopropyl adipate, di-2-ethylhexyl sebacate, cetyl lactate, apple Diisostearyl acid, ethylene glycol di-2-ethylhexanoate, neopentyl glycol dicaprate, glycerin di-2-heptylundecanoate, glycerin tri-2-ethylhexanoate, trimethylolpropane tri-2-ethylhexanoate, Oil agents such as synthetic ester oils such as trimethylolpropane triisostearate and pentane erythritol tetra-2-ethylhexanoate; fatty acid soap (sodium laurate) Anionic surfactants such as potassium lauryl sulfate and alkylsulfuric triethanolamine ether; cationic surfactants such as stearyltrimethylammonium chloride, benzalkonium chloride and laurylamine oxide; imidazoline-based amphoteric Amphoterics such as surfactants (2-cocoyl-2-imidazolinium hydroxide-1-carboxyethyloxy disodium salt, etc.), betaine surfactants (alkyl betaine, amide betaine, sulfobetaine, etc.), acylmethyl taurine, etc. Surfactants: sorbitan fatty acid esters (such as sorbitan monostearate and sorbitan sesquioleate), glycerin fatty acids (such as glyceryl monostearate), propylene glycol fatty acid esters (monostearin) Propylene glycol acid), hydrogenated castor oil derivative, glycerin alkyl ether, POE sorbitan fatty acid esters (POE sorbitan monooleate, polyoxyethylene sorbitan monostearate, etc.), POE sorbite fatty acid esters (POE-sorbite monolaurate, etc.) ), POE glycerin fatty acid esters (such as POE-glycerin monoisostearate), POE fatty acid esters (such as polyethylene glycol monooleate and POE distearate), POE alkyl ethers (such as POE2-octyldodecyl ether), POE alkylphenyl ether (POE nonylphenyl ether, etc.), Pluronic type, POE / POP alkyl ethers (POE / POP2-decyltetradecyl ether, etc.), Tet Nicks, non-ionic surfactants such as POE castor oil / hardened castor oil derivatives (POE castor oil, POE hardened castor oil, etc.), sucrose fatty acid esters, alkyl glucosides; polyethylene glycol, glycerin, erythritol, sorbitol, xylitol, Polyhydric alcohols such as maltitol, propylene glycol and 2,4-hexanediol; moisturizing ingredients such as sodium pyrrolidone carboxylate, lactic acid and sodium lactate; paraaminobenzoic acid ultraviolet absorbers; anthranilic acid ultraviolet absorbers; salicylic acid -Based UV absorbers; cinnamic acid-based UV absorbers; benzophenone-based UV absorbers; sugar-based UV absorbers; 2- (2'-hydroxy-5'-t-octylphenyl) benzotriazole, 4-methoxy-4'- UV such as t-butyldibenzoylmethane Absorbing agents; ethanol, lower alcohols antibacterial agents such phenoxyethanol such isopropanol can be preferably exemplified.

Oral preparations include, for example, general foods such as confectionery, bread and noodles, drink preparations, food groups with the purpose of promoting health in the form of capsules and tablets (for example, foods for specified health use), granules, powders Examples of such drugs include capsules, capsules, orally administered drugs in the form of tablets.
In the case of an oral dosage form, it can contain any optional component that is acceptable. Examples of such optional ingredients include foods such as salt, sugar, sodium glutamate, sodium inosinate, vinegar and other flavoring ingredients, coloring ingredients, flavoring ingredients such as flavors, thickeners, emulsifying / dispersing agents, and preservatives. Stabilizers, various vitamins and the like can be preferably exemplified, and if it is a food group or pharmaceutical product having the purpose of promoting health, excipients such as crystalline cellulose and lactose, binders such as arabic gum and hydroxypropylcellulose, croscarm Preferred examples include disintegrants such as loin sodium and starch, lubricants such as magnesium stearate, flavoring agents, flavoring agents, coloring agents, various vitamins and the like. By treating these according to a conventional method, the composition for oral administration of the present invention can be produced.

The total content of the plant extract in the oral preparation can be 0.05 to 100% by mass, more preferably 30 to 80% by mass, as a solid content.
In addition, it is preferable to take 10 to 1000 mg of the plant extract as a solid content in one or several divided doses.

  The lysosomal activity promoter of the present invention can be used for the treatment or prevention of various diseases caused by a decrease in lysosomal activity. Examples of such diseases include lysosomal storage diseases including lipid storage disease, mucopolysaccharidosis, mucolipidosis and glycoprotein storage disease, and more specifically, activator deficiency / GM2 gangliosidosis, α- Mannosidosis, aspartylglucosamineuria, cholesterol ester accumulation disease, chronic hexosaminidase A deficiency, cystinosis, Danone disease, Fabry disease, Faber disease, fucosidosis, galactosialidosis, Gaucher disease (eg, type I, II Type III), GM1 gangliosidosis (eg, infantile, late infantile / juvenile, adult / chronic), I-cell disease / mucolipidosis II, infantile free sialic acid accumulation / ISSD, juvenile hex Sosaminidase A deficiency, Krabbe disease (eg infantile onset, delayed onset), metachromatic Dystrophies, mucopolysaccharidosis, pseudo-Harler polydystrophy / mucolipidosis IIIA (eg, MPSI Hurler syndrome, MPSI Scheyer syndrome, MPS I Hurler Scheyer syndrome, MPS II Hunter syndrome, San Philippo syndrome type A / MPS III A, San Filippo syndrome type B / MPS III B, San Filippo syndrome type C / MPS III C, San Filippo syndrome type D / MPS III D, Morquio type A / MPS IVA, Morquio type B / MPS IVB, MPS IX hyaluronidase deficiency, MPS VI Marlotho Lamy, MPS VII Sly syndrome, mucolipidosis I / sialidosis, mucolipidosis IIIC, mucolipidosis type IV), multiple sulfatase deficiency, Niemann Pi Cook's disease (eg, A, B, C), neuronal ceroid lipofuscinosis (eg, CLN6 disease-atypical late infantile, late onset, early juvenile, Batten Spielmeier Vogt / young Sexual NCL / CLN3 disease, Finnish variant of late infantile CLN5, Jansky Beershawsky disease / late infantile CLN2 / TPP1 disease, Kuchs / adult-onset NCL / CLN4 disease, Northern epilepsy / late infantile CLN8 variant, Santavori-Haltia / infant CLN1 / PTT disease, β-mannosidosis), Pompe disease / glucogen storage disease type II, concentrated dystrophy, Sandhoff disease / GM2 ganglioside Cis (eg, adult onset, infanthood, juvenile), Schindler disease, Sarah disease / Sialic acid storage disease, Tay-Sachs disease / GM2 gangliosidosis or Wolman disease.

The lysosome activity promoter of the present invention can be used for skin whitening.
Further, it is known that the amount of fragmented collagen is significantly higher in the skin of elderly people, and lysosomes are known to have the resolution of this fragmented collagen. Hyaluronic acid is a substance that is very rapidly metabolized by nature, but the ability to metabolize hyaluronic acid decreases because the degradation function of lysosomes stagnates with aging. Therefore, by improving the activity of the lysosome, it is possible to normalize the metabolism of the fragmented collagen accumulated by aging and the hyaluronic acid whose metabolic ability has decreased by aging.
That is, the lysosome activity promoter of the present invention can be used for anti-aging.

<2> Hyaluronic Acid Production Promoter The hyaluronic acid production promoter of the present invention is an extract of a plant belonging to the genus Poaceae (Coix) and / or a plant belonging to the family Betulaceae (Betula). The embodiment thereof is the same as the embodiment of the invention relating to the above-mentioned lysosome activity promoter.
Moreover, the hyaluronic acid production promoter of the present invention can be used for improving skin elasticity, moisture retention, and anti-aging.

<3> Screening method The present invention also relates to a screening method for a component having hyaluronic acid production promoting action, characterized by using lysosomal activity as an index.
The activity of lysosome is positively correlated with the amount of hyaluronic acid produced. Therefore, if the activity of the lysosome is improved by exposing the cells to the test sample, it can be determined that the test sample has a hyaluronic acid production improving effect.

  In the screening method of the present invention, experimental animals may be used, but cultured cells are preferably used. Specifically, an embodiment in which lysosomal activity in cultured cells after adding a test sample to the medium is preferably measured.

A cell line may be used as the cultured cell, or a primary cultured cell may be used.
The cell type is not particularly limited, but epidermal cells and fibroblasts are preferably used.

The method for measuring lysosomal activity is not particularly limited, and a commercially available kit may be used. As a commercially available kit, Cell Navigator ^™ Lysosome Staining Kit (AAT Bioquest) and the like can be exemplified.

<Plant extract>
The pearl barley extract used in the examples was prepared by the following method. That is, 2 kg of seeds (Yokuinin) excluding the seed coats of commercially available Gramineae genus pearl barley were heated and refluxed twice for 2 hours in 9 liters of methanol, and after filtration, the filtrate was combined for 2 times. Concentrated and dried to obtain a Yokuinin methanol extract.
On the other hand, as the birch extract used in the examples, a commercially available product (birch extract: manufactured by Maruzen Pharmaceutical Co., Ltd.) was prepared.

<Test Example 1>
Normal human dermal fibroblasts were seeded in 96-well plates at 5.0 × 10 ³ cells / well and cultured at 37 ° C. in a 5% CO ₂ environment until the cells adhered. Plant extracts were added to each well of the plate seeded with cells at the final concentrations and combinations shown in FIG. 1 and Table 1. Only the solvent was added to the control. Thereafter, the lysosome and the nucleus were fluorescently labeled with a lysosome staining solution (Cell Navigator ^™ Lysosome Staining Kit (AAT Bioquest)) and a nuclear staining solution (Hoechst), respectively, and the fluorescence intensity was measured. The value divided by (number of cells) was used as the amount of lysosome per cell, and calculated as a relative value when the amount of lysosome in the control was 1. The results are shown in FIG.

As shown in FIG. 1 and Table 1, the fluorescence intensity of lysosomes in cultured cells to which a pearl barley extract or birch extract was added to the medium is significantly higher than that of the control. This result shows that the pearl barley extract and the birch extract have the action of promoting the activity of lysosomes.
In addition, the fluorescence intensity of lysosomes in cultured cells in which a mixture of pearl barley extract and birch extract was added to the medium was significantly higher than when each extract was added alone to the medium (FIG. 1 and Table 1). ). Considering that the final concentration in the medium of the mixture of the two extracts is low compared to when each extract was added to the medium alone, this result indicates that the pearl barley extract and birch extract have lysosomal activity It shows that a synergistic effect is shown in the promoting action of.

<Test Example 2>
Using normal human epidermal keratinocytes and normal human dermal fibroblasts, the lysosomal activity promoting effect of the extract at the final concentrations and combinations shown in FIGS. It was measured by. The results are shown in FIGS.

  As shown in FIGS. 2 and 3 and Tables 2 and 3, similar to the results of Test Example 1, the mixture of pearl barley extract and birch extract has the effect of promoting the activity of lysosomes.

<Test Example 3>
Normal human epidermal keratinocytes and normal human dermal fibroblasts were seeded in a 24-well plate at 5 × 10 ⁴ cells / well and cultured at 37 ° C. in a 5% CO ₂ environment until the cells adhered. Plant extracts were added to the respective wells of the plates seeded with cells at the final concentrations and combinations shown in FIGS. Only the medium was added to the control. After overnight culture at 37 ° C. and 5% CO ₂ , 350 μl of RLT Buffer (RNeasy Mini Kit, QIAGEN) was added to collect the cultured cells, and RNA was extracted using RNeasy Mini Kit. The extracted RNA was reverse transcribed using SuperScript VILO cDNA Synthesis Kit to synthesize cDNA, and the gene expression level of hyaluronic acid synthase in epidermal cells and fibroblasts was analyzed by real-time qPCR using QuantFast SYBR Green PCR Kit . The relative value when the expression level of hyaluronic acid synthase in the control was 1 was calculated. The results are shown in Tables 4 and 5 and FIGS.

As shown in Tables 4 and 5 and FIGS. 4 and 5, the gene expression level of hyaluronic acid synthase in the cultured cells to which the pearl barley extract or the birch extract was added to the medium is significantly higher than that of the control. This result shows that the pearl barley extract and the birch extract have the effect of improving the hyaluronic acid production ability.
Also, looking at the results of gene expression levels of hyaluronic acid synthase after overnight culture after adding the extract to the medium, the expression level for the control when each extract was added to the medium alone was extracted from birch. The highest is the highest, followed by the gentian extract, but the rate of increase when adding a mixture of barley extract and birch extract to the medium is greater than the rate of increase when these mixtures are added to the medium. Remarkably high. This result indicates that the pearl barley extract and the birch extract exhibit a remarkable synergistic effect in improving the hyaluronic acid production ability.

<Test Example 4>
Normal human epidermal keratinocytes and normal human dermal fibroblasts were seeded in a 24-well plate at 5 × 10 ⁴ cells / well and cultured at 37 ° C. in a 5% CO ₂ environment until the cells adhered. A mixture of Leupeptin and Pepstatin, which are lysosomal activity inhibitors, was added to each well of the plate seeded with cells so that the final concentration in the medium was 100 uM. Only the medium was added to the control. After culturing at 37 ° C. in a 5% CO ₂ environment for 24 hours, the amount of hyaluronic acid produced in epidermal cells and fibroblasts was analyzed by the same method as in Test Example 3. The amount of hyaluronic acid produced when the amount of hyaluronic acid produced in the control was 1 was calculated. The results are shown in Tables 6 and 7 and FIGS.

  As shown in Tables 6 and 7 and FIGS. 6 and 7, it was found that the amount of hyaluronic acid produced in epidermal cells and fibroblasts was reduced by inhibiting the activity of lysosomes. This result suggests that lysosomal activity affects the amount of hyaluronic acid produced.

  Combining the results of Test Example 3 and Test Example 4, it is shown that the pearl barley extract and the birch extract have the effect of improving the hyaluronic acid production ability by improving the lysosomal activity.

  The present invention can be applied to cosmetics and supplements.

Claims (7)

  A lysosomal activity promoter comprising an extract of a plant belonging to the genus Poaceae (Coix) and / or an extract of a plant belonging to the genus Betulaceae (Betula).   The lysosomal activity promoter according to claim 1, comprising a mixture of an extract of a plant belonging to the genus Poaceae (Coix) and an extract of a plant belonging to the genus Betulaceae (Betula).   The lysosomal activity promoter according to claim 1 or 2, wherein the plant belonging to the genus Poaceae (Coix) is barley (Coix lacryma-jobi var. Ma-yuen).   The lysosomal activity promoter according to any one of claims 1 to 3, wherein the plant belonging to the family Betulaceae (Betula) is birch (Betula platyphylla).   The lysosomal activity promoter according to any one of claims 1 to 4, which is used for promoting production of hyaluronic acid.   The lysosome activity promoter according to any one of claims 1 to 5, which is an external preparation or an oral preparation. A screening method for a component having a hyaluronic acid production promoting action, characterized by using lysosomal activity as an index.