Lysosome activity promoter

Abstract

To provide a novel lysosome activity promoter.SOLUTION: A lysosome activity promoter of a fibroblast has an extract from a plant belonging to Apiaceae Angelica as an active ingredient.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a lysosome activity promoter in fibroblasts.

Background Art

[0002] Lysosomes are one of the organelles of eukaryotes and are sites of intracellular digestion with hydrolytic enzymes inside. Biopolymers taken into the membrane by endocytosis or autophagy are decomposed in lysosomes. Among the decomposition products, useful ones are absorbed into the cytoplasm and reused, and waste products are discarded outside the cell by exocytosis or remain inside the cell as residual bodies.

[0003] It is known that lysosome activity is involved in various diseases. As a typical disease, lysosomal storage diseases are known. Lysosomal storage diseases often develop due to the deficiency or mutation of hydrolytic enzymes acting in lysosomes, resulting in the accumulation of the substrates of the enzymes as deposits inside cells without being decomposed. Lysosomal storage diseases have a wide range of clinical heterogeneity and biological diversity, and include lipid storage diseases, mucopolysaccharidoses, mucolipidoses, and glycoprotein storage diseases (see Patent Documents 1 and 2).

[0004] Therefore, in recent years, searches have been made for components that improve lysosome activity, and extracts of plants belonging to the genus Coix of the family Poaceae and extracts of plants belonging to the genus Betula of the family Betulaceae are known to promote lysosome activity (Patent Document 3).

[0005] On the other hand, it is known that a whitening effect can be obtained by improving lysosome activity in keratinocytes and promoting the degradation of melanosome proteins (Patent Document 4, Non-Patent Documents 1 and 2).

[0006] In addition, it is known that cathepsin B1 and D, which are lysosomal enzymes, degrade collagen (Non-Patent Document 3), and that hyaluronic acid is taken up and degraded in lysosomes (Non-Patent Document 4).

Prior Art Documents

Patent Documents

[0007]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Non-Patent Documents

[0008]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Summary of the Invention

Problems to be Solved by the Invention

[0009] As described above, lysosomes are associated with various diseases, and the development of components that can improve their activity is desired. Therefore, an object of the present invention is to provide a novel lysosome activity promoter.

Means for Solving the Problems

[0010] The present invention for solving the above problems is a promoter for lysosome activity of fibroblasts, which contains an extract of a plant belonging to the genus Angelica of the family Apiaceae as an active ingredient. The promoter for lysosome activity of fibroblasts of the present invention is useful for the treatment or prevention of diseases caused by a decrease in lysosome activity, and for improving biological functions related to lysosome activity, particularly for improving skin conditions.

[0011] In a preferred form of the present invention, the plant belonging to the genus Angelica of the family Apiaceae is Angelica keiskei.

[0012] In a preferred form of the present invention, the present invention is a promoter for lysosome activity for promoting the lysosome activity of the dermis.

[0013] In a preferred form of the present invention, the content of the extract of the plant is 0.5 to 4.0% by mass based on the dry mass.

[0014] The present invention also relates to an external preparation for promoting the lysosome activity of fibroblasts, which contains the above-mentioned promoter for lysosome activity of fibroblasts. Such an external preparation of the present invention can be easily used continuously and is useful for the treatment or prevention of diseases caused by a decrease in lysosome activity, improvement of biological functions related to lysosome activity, particularly improvement of skin conditions.

Advantages of the Invention

[0015] The lysosome activity promoter for fibroblasts of the present invention can improve the activity of lysosomes in fibroblasts.

Brief Description of the Drawings

[0016]

Figure 1

Figure 2

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Embodiments for Carrying Out the Invention

[0017] The fibroblast lysosome activity promoter of the present invention (hereinafter simply referred to as the lysosome activity promoter) contains an extract of a plant belonging to the genus Angelica of the family Apiaceae (hereinafter also simply referred to as the extract of the genus Angelica). As the plant belonging to the genus Angelica of the family Apiaceae, Angelica keiskei is preferably mentioned.

[0018] The extraction site of the plant belonging to the genus Angelica can be appropriately selected according to the purpose. For example, the leaf part, stem part, flower part, fruit part, root part, etc. can be mentioned. Among these, it is particularly preferable to use the leaf part and / or the stem part.

[0019] At the time of extraction, it is preferable to process the extraction site or its dried product of the plant belonging to the genus Angelica in advance by pulverizing or cutting it finely to improve the extraction efficiency. The drying of the extraction site may be carried out in the sun or using a commonly used dryer.

[0020] Examples of the extraction method include immersing the extraction site in an extraction solvent under normal pressure, or under pressure or reduced pressure, at room temperature, cooled or heated, and extracting, using a distillation method such as steam distillation, and a pressing method of pressing the extraction site to obtain an extract. These methods can be used alone or in combination of two or more for extraction.

[0021] When extracting by immersion, 1 to 30 parts by mass of the solvent is added to 1 part by mass of the dried substance of the plant body, above-ground part, rhizome part, and / or seeds of the plant belonging to the genus Angelica, and it can be carried out by immersing for several days at room temperature or for several hours at a temperature near the boiling point of the solvent. After immersion, it is cooled to room temperature, and after removing unnecessary substances, a concentrated solution can be obtained by concentrating the solvent under reduced pressure or the like. By allowing the concentrated solution to stand at 5°C and filtering the precipitated precipitate, the desired extract can be obtained.

[0022] As the extraction solvent, polar solvents are preferred, and one or more selected from alcohols such as water, methanol, ethanol, propyl alcohol, isopropyl alcohol, and butanol, polyhydric alcohols such as 1,3-butylene glycol, polypropylene glycol, and glycerin, ketones such as acetone and methyl ethyl ketone, and ethers such as diethyl ether and tetrahydrofuran can be preferably exemplified. Among them, it is preferable to extract using 1,3-butylene glycol or water. The extract thus obtained, or the appropriately concentrated one, can be used as the lysosome activity promoter of the present invention.

[0023] In addition, commercially available extracts of the genus Sicyos can be used, and these can be used as the lysosome activity promoter of the present invention. For example, "Ashitaba Extract BG (manufactured by Maruzen Pharmaceutical Co., Ltd.)" can be used.

[0024] Based on the dry mass, the content of the extract of the genus Sicyos in the total amount of the lysosome activity promoter of the present invention is preferably 0.5 to 4.0% by mass, more preferably 0.9 to 2.7% by mass, and still more preferably 1.5 to 2.0% by mass.

[0025] The lysosome activity promoter of the present invention is excellent in the effect of promoting the activity of lysosomes in fibroblasts. Fibroblasts are one of the cells that make up connective tissue and are responsible for synthesizing components that make up extracellular matrix such as collagen, elastin, and hyaluronic acid. Fibroblasts are present in skin tissues such as the dermis, subcutaneous tissues such as skin ligaments, muscle tissues, and nerve tissues. The lysosome activity promoter of the present invention can be used to promote the lysosome activity of tissues where the above fibroblasts are distributed, and preferably can be used to promote the lysosome activity of fibroblasts in the dermis.

[0026] The lysosome activity promoter of the present invention may be used as a stock solution, or may be diluted to any concentration before use. Further, the lysosome activity promoter of the present invention can be formulated into a topical preparation or an oral preparation by appropriately combining it with any component used in formulation. In the present invention, it is preferable to form a topical preparation or an oral preparation containing the lysosome activity promoter.

[0027] Examples of external preparations include cosmetics, quasi-drugs, pharmaceuticals, etc., and they can also contain any components that are usually used, as long as the effects of the present invention are not impaired. Examples of such optional components include oils and waxes such as 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, hydrogenated coconut oil, hydrogenated oil, beeswax, hydrogenated castor oil, honey, candelilla wax, carnauba wax, spermaceti wax, lanolin, reduced lanolin, hard lanolin, jojoba wax, etc.; hydrocarbons such as liquid paraffin, squalane, pristane, ozokerite, paraffin, ceresin, petrolatum, microcrystalline wax, etc.; higher fatty acids such as oleic acid, isostearic acid, lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, undecylenic acid, etc.; higher alcohols such as cetyl alcohol, stearyl alcohol, isostearyl alcohol, behenyl alcohol, octyldodecanol, myristyl alcohol, cetostearyl alcohol, etc.; oil agents such as synthetic ester oils such as cetyl isooctanoate, isopropyl myristate, hexadecyl isostearate, diisopropyl adipate, di-2-ethylhexyl sebacate, cetyl lactate, diisostearyl malate, ethylene glycol di-2-ethylhexanoate, neopentyl glycol dicaprate, glycerin di-2-heptylundecanoate, glycerin tri-2-ethylhexanoate, trimethylolpropane tri-2-ethylhexanoate, trimethylolpropane triisostearate, pentaerythritol tetra-2-ethylhexanoate, etc.; anionic surfactants such as fatty acid soaps (sodium laurate, sodium palmitate, etc.), potassium lauryl sulfate, alkyl sulfate triethanolamine ether, etc.; cationic surfactants such as stearyl trimethylammonium chloride, benzalkonium chloride, laurylamine oxide, etc.; amphoteric surfactants such as imidazoline-based amphoteric surfactants (2-cocoyl-2-imidazolinium hydroxide-1-carboxyethyloxy 2 sodium salt, etc.), betaine-based surfactants (alkyl betaine, amide betaine, sulfobetaine, etc.), acylmethyl taurine, etc.Sorbitan fatty acid esters (such as sorbitan monostearate, sorbitan sesquioleate, etc.), glycerin fatty acids (such as glycerin monostearate, etc.), propylene glycol fatty acid esters (such as propylene glycol monostearate, etc.), hydrogenated castor oil derivatives, glycerin alkyl ethers, POE sorbitan fatty acid esters (such as POE sorbitan monooleate, polyoxyethylene sorbitan monostearate, etc.), POE sorbit fatty acid esters (such as POE-sorbitan monolaurate, etc.), POE glycerin fatty acid esters (such as POE-glycerin monoisostearate, etc.), POE fatty acid esters (such as polyethylene glycol monooleate, POE distearate, etc.), POE alkyl ethers (such as POE2-octyldodecyl ether, etc.), POE alkyl phenyl ethers (such as POE nonyl phenyl ether, etc.), pluronic (registered trademark) types, POE·POP alkyl ethers (such as POE·POP2-decyltetradecyl ether, etc.), tetronic types, POE castor oil·hydrogenated castor oil derivatives (such as POE castor oil, POE hydrogenated castor oil, etc.), sucrose fatty acid esters, nonionic surfactants such as alkyl glucoside; polyhydric alcohols such as polyethylene glycol, glycerin, erythritol, sorbitol, xylitol, maltitol, propylene glycol, 2,4-hexanediol; moisturizing components such as sodium pyrrolidone carboxylate, lactic acid, sodium lactate; para-aminobenzoic acid-based ultraviolet absorbers; anthranilic acid-based ultraviolet absorbers; salicylic acid-based ultraviolet absorbers; cinnamic acid-based ultraviolet absorbers; benzophenone-based ultraviolet absorbers; sugar-based ultraviolet absorbers; ultraviolet absorbers such as 2-(2'-hydroxy-5'-t-octylphenyl) benzotriazole, 4-methoxy-4'-t-butyldibenzoylmethane; lower alcohols such as ethanol, isopropanol; antibacterial agents such as phenoxyethanol, etc. can be preferably exemplified.;

[0028] Examples of oral preparations include general foods such as confectionery, bread, noodles, etc., drink preparations, food groups for health promotion in the form of capsules or tablets (such as foods for specified health use, etc.), granules, powders, capsules, and oral pharmaceuticals in the form of tablets, etc. When in the form of an oral preparation, it can contain acceptable optional components. Such optional components, if it is a food, can preferably include flavoring components such as salts, sugars, sodium glutamate, sodium inosinate, vinegar, coloring components, odor-masking components such as flavors, thickeners, emulsifying / dispersing agents, preservatives, stabilizers, various vitamins, etc. If it is a food group or a pharmaceutical product having the purpose of promoting health, excipients such as crystalline cellulose and lactose, binders such as gum arabic and hydroxypropyl cellulose, disintegrants such as croscarmellose sodium and starch, lubricants such as magnesium stearate, flavoring and odor-masking agents, coloring agents, various vitamins, etc. can be preferably exemplified. By treating these according to conventional methods, the oral preparation of the present invention can be produced.

[0029] The total content of the extract of the genus Schisandra in the external preparation is 0.0005 to 0.0040% by mass, more preferably 0.0009 to 0.0027% by mass, as the dry mass. The total content of the extract of the genus Schisandra in the oral preparation can be 0.05 to 100% by mass, more preferably 30 to 80% by mass, as the dry mass. Also, it is preferable to take the extract of the plant in the form of 10 to 1000 mg per day as the solid content, divided into one or several times for drinking.

[0030] The lysosome activity promoter of the present invention can be used for the treatment or prevention of various diseases caused by the decrease in lysosome activity. Such diseases include lysosomal storage diseases including lipid storage diseases, mucopolysaccharidoses, mucolipidoses, and glycoprotein storage diseases. More specifically, activator deficiency / GM2 gangliosidosis, α-mannosidosis, aspartylglucosaminuria, cholesterol ester storage disease, chronic hexosaminidase A deficiency, cystinosis, Danon disease, Fabry disease, Farber disease, fucosidosis, galactosialidosis, Gaucher disease (e.g., type I, type II, type III), GM1 gangliosidosis (e.g., infantile, late infantile / juvenile, adult / chronic), I cell disease / mucolipidosis II, infantile free sialic acid storage disease / ISSD, juvenile hexosaminidase A deficiency, Krabbe disease (e.g., infantile onset, delayed onset), metachromatic leukodystrophy, mucopolysaccharidosis diseases, pseudo-Hurler polydystrophy / mucolipidosis IIIA (e.g., MPS I Hurler syndrome, MPS I Scheie syndrome, MPS I Hurler-Scheie syndrome, MPS II Hunter syndrome, Sanfilippo syndrome type A / MPS III A, Sanfilippo syndrome type B / MPS III B, Sanfilippo syndrome type C / MPS III C, Sanfilippo syndrome type D / MPS III D, Morquio type A / MPS IV A, Morquio type B / MPS IV B, MPS IX hyaluronidase deficiency, MPS VI Maroteaux-Lamy, MPS VII Sly syndrome, mucolipidosis I / sialidosis, mucolipidosis III C, mucolipidosis type IV), multiple sulfatase deficiency, Niemann-Pick disease (e.g., type A, type B, type C), neuronal ceroid lipofuscinosis (e.g., CLN6 disease - atypical late infantile, late-onset variant, early juvenile, Batten-Spielmeyer-Vogt / juvenile NCL / CLN3 disease, Finnish variant of late infantile CLN5, Jansky-Bielschowsky disease / late infantile CLN2 / TPP1 disease, Kufs / adult-onset NCL / CLN4 disease, northern epilepsy (NorthernEpilepsy) / variant of late infantile CLN8, Santavuori-Haltia / infantile CLN1 / PTT disease, β-mannosidosis), Pompe disease / glycogen storage disease type II, osteopetrosis condensans, Sandhoff disease / GM2 gangliosidosis (e.g., adult-onset, infantile, juvenile), Schindler disease, Salla disease / sialic acid storage disease, Tay-Sachs disease / GM2 gangliosidosis or Wolman disease, etc.

[0031] The lysosome activity promoter of the present invention can be used for improving skin conditions caused by a decrease in lysosome activity. Specifically, it can be used for promoting hyaluronic acid production or promoting collagen production.

Examples

[0032] <Test Example 1> Evaluation of lysosome activity in fibroblasts using an extract of Japanese butterbur Normal human dermal fibroblasts (NHDF, #5884 neonate) suspended in medium (10% FBS + 1% AA / DMEM) were dispensed into 4-well chambers at 2.5×104 cells / well and cultured in an environment of 37°C and 5% CO2 for 24 hours. Japanese butterbur extract (Japanese butterbur extract BG (manufactured by Maruzen Pharmaceutical Co., Ltd.)) was added to fresh medium to a final concentration of 0.1% or 0.05% (0.00148% and 0.00074% by dry mass, respectively) to prepare an extract-containing medium. For the cells after culture, the medium in the chamber was replaced with the extract-containing medium and cultured in an environment of 37°C and 5% CO2 for 24 hours. As a control, a medium without Japanese butterbur extract added as the medium for replacement after culture was used and cultured under the same conditions.

[0033] After culture, the medium was removed, the cells were washed with PBS, and then 500 μl / well of the following staining medium was added and cultured at 37°C for 30 minutes or more. · Staining medium: Diluted 10,000-fold with the medium prepared by adding 10 μl of DMSO to 1 unit of EsterGreen in Lysosomal Metabolic Health Assay Kit (Marker Gene Technologies, Inc.)

[0034] The medium was removed, washed with PBS, and then 500 μl / well of 5×Opti-Klear diluted 5-fold with sterile water was added to prepare cells for observation. Subsequently, cells were observed under a fluorescence microscope at a measurement wavelength of EX / EM: 490 / 520 nm. The results are shown in Figures 1 to 3.

[0035] As shown in Figures 1 to 3, when 0.1% (Figure 2) and 0.05% (Figure 3) of the Houttuynia cordata extract were added, the fluorescence intensity of lysosomes increased compared to the control without the extract addition (Figure 1). Therefore, it was revealed that the Houttuynia cordata extract has an effect of promoting lysosomal activity in fibroblasts.

[0036] <Test Example 2> Evaluation of lysosomal activity in fibroblasts using a lotion containing Houttuynia cordata extract as an active ingredient (1) Preparation of lotion According to the following formulation, a cosmetic, which is a topical skin preparation, was prepared. That is, the formulation components were heated to 80°C, stirred and solubilized, and then stirred and cooled to obtain the lotion of Example 1. Also, Comparative Example 1 (without addition of Houttuynia cordata extract BG) was prepared by replacing an equal amount of water with the following Houttuynia cordata extract BG.

[0037]

Table 1

[0038] (2) Evaluation test of lysosomal activity Normal human dermal fibroblasts (NHDF, #5884 neonatal) suspended in medium (10% FBS + 1% AA / DMEM) were dispensed into a 4-well chamber at 2.5×104 cells / well and cultured for 24 hours in an environment of 37°C and 5% CO2. The Example 1 or Comparative Example 1 was added to a fresh medium to a 40-fold dilution, and these were used as lotion-containing media. Regarding the cells after culture, the medium in the chamber was replaced with the lotion-containing medium, and the cells were cultured at 37°C in a 5% CO2 environment for 24 hours.

[0039] After culture, the medium was removed, and after washing with PBS, 500 μl / well of the following staining medium was added, and the cells were cultured at 37°C for 30 minutes or more. · Staining medium: 10 μl of DMSO was added to 1 unit of EsterGreen of Lysosomal Metabolic Health Assay Kit (Marker Gene Technologies, Inc.), and the mixture was diluted 10,000-fold with the above medium

[0040] The medium was removed, and after washing with PBS, 5×Opti-Klear diluted 5-fold with sterile water was added at 500 μl / well to prepare cells for observation. Then, the cells were observed with a fluorescence microscope at a measurement wavelength of EX / EM: 490 / 520 nm. The results are shown in FIGS. 4 and 5.

[0041] As shown in FIGS. 4 and 5, when Example 1 containing the ashitaba extract was added (FIG. 5), the fluorescence intensity of lysosomes increased compared to the case where Comparative Example 1 not containing the extract was added (FIG. 4). Therefore, it was clarified that the lotion containing the ashitaba extract has an effect of promoting lysosomal activity in fibroblasts.

Industrial Applicability

[0042] The present invention can be applied to pharmaceuticals, cosmetics, supplements, etc.

Claims

【Claim 1】 A fibroblast lysosome activity promoter containing an extract of a plant belonging to the genus Angelica of the family Apiaceae as an active ingredient.

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