JP5649995B2 - Ceramide production promoter - Google Patents

Description

  The present invention relates to a ceramide production promoter containing at least one extract selected from the group consisting of Diou and Gennosho extract.

  Ceramide is a kind of sphingolipid and is a general term for a group of compounds in which sphingosine and a fatty acid are amide-bonded. Ceramide is known to be present at a high concentration in the cell membrane. Ceramide is a component of keratin intercellular lipids, and is secreted extracellularly in the process of keratinization, and plays an important role in the skin barrier function and water retention ability. Furthermore, ceramide is known as a signal transducing substance to control cell proliferation, differentiation, apoptosis, and the like. From these facts, substances that promote ceramide production can be expected to suppress cell proliferation, induce differentiation and induce apoptosis, and can be expected to have therapeutic effects on diseases caused by these abnormalities.

  Examples of skin diseases highly related to ceramide include squamous cell carcinoma, psoriasis, and atopic dermatitis.

  Squamous cell carcinoma is a type of epithelial malignant tumor, which occurs when epidermal keratinocytes become malignant and over proliferate. So far, it is known that glycosylceramide, which is a derivative of ceramide, reduces squamous cell carcinoma (Non-patent Document 1). As its mechanism, ceramide induces apoptosis in cancer cells and suppresses the growth of cancer cells (Non-patent Document 2). Furthermore, it contains a cancer metastasis inhibitor characterized by containing a ceramide derivative (Patent Document 1), a ceramide production promoter for the treatment of malignant tumors (Patent Document 2), and a ceramide as an antitumor active substance. Liposomes (Patent Document 3) and the like characterized by this are disclosed. From the above, promotion of ceramide production is indispensable for suppressing the growth and metastasis of squamous cell carcinoma in the skin.

  Psoriasis is a chronic skin keratosis disease that is thought to be affected by a genetic predisposition. In the lesioned part, the proliferation of epidermal keratinocytes is increased compared with the non-lesioned part, and a decrease in the barrier function and a decrease in the amount of ceramide are observed (Non-patent Document 3). Also disclosed are ceramide production promoters (patent documents 4 to 6) characterized by containing active ingredients derived from genquanine, camellia plants, and Lamiaceae plants for the purpose of treating and preventing psoriasis. Yes.

  Atopic dermatitis is a chronic skin disease accompanied by itching, and many people with allergies are affected. It is known that the amount of ceramide is reduced in the skin of patients with atopic dermatitis compared to healthy skin (Non-patent Document 4), and the lack of ceramide in the stratum corneum is a factor in atopic dermatitis. It is considered. Further, it is characterized by comprising as an active ingredient a ceramide-containing composition for the treatment of atopic dermatitis (Patent Document 7), a glycosphingolipid or α-D-glucopyranosylglycerols supplying ceramide. An atopic dermatitis therapeutic agent (Patent Documents 8 to 9) and the like are disclosed.

  From these facts, it is possible to treat and prevent skin diseases such as squamous cell carcinoma, psoriasis and atopic dermatitis by promoting the production of ceramide in the skin.

  Moreover, it is known that the gentian extract has an effect of promoting blood flow (Patent Document 10), and the gentian extract has an active oxygen scavenging effect (Patent Document 11). However, the ceramide production promoting effect of Diou and Gennoshoco extract has not been reported.

JP-A-11-343249 JP 2010-70499 JP-A-9-110722 JP2004-161648 JP-A-2004-189683 JP2004-210743A JP 2000-119178 A JP 2003-231640 A JP 2009-161564 A JP 2006-206513 A JP2003-300858

Kazunori Fujiwara, Bulletin of the Japan Otolaryngology Society, 113: 343, 2010 David E, Mol Cancer Ther, 5: 200-208,2006 Cho Y, J Korean Med Sci, 19: 859-863, 2004 Sasakawa Genso, Oil Chemistry, 44: 751-766, 1995

  An object of the present invention is to provide a ceramide production promoter having a high ceramide production promotion effect.

  As a result of intensive studies, the present inventors have found an excellent ceramide production promoting effect in the extract of Gera and Gennosho, and have completed the present invention.

  The present invention is a ceramide production promoter containing at least one extract selected from the group consisting of Diou and Gennosho extract.

  The present invention is a therapeutic agent for squamous cell carcinoma comprising at least one extract selected from the group consisting of Diou and Gennosho extract.

  The present invention is a therapeutic agent for psoriasis containing at least one extract selected from the group consisting of Diou and Gennosho extract.

  The present invention is a therapeutic agent for atopic dermatitis comprising at least one extract selected from the group consisting of Diou and Gennosho extract.

  The Diou and Gennosho extract of the present invention increases the amount of ceramide in keratinocyte lipids by promoting production of ceramide in epidermal keratinocytes, and is highly associated with squamous cell carcinoma, psoriasis and atopy. It is possible to treat and prevent skin diseases such as atopic dermatitis.

  The Giant used in the present invention belongs to the genus Giantiaceae, and is a plant of scientific name: Rehmannia glutinosa (Gearth.) Libosch. Perennial plants native to China, whose rhizomes are dried (herbal medicine name: natural yellow) can be used.

  Genus Dwarf used in the present invention belongs to the genus Fusoso, and is a plant having the scientific name: Geranium thunbergii. A perennial that grows naturally in various parts of Japan, the Korean Peninsula, and the mainland of China. Dry roots, stems, leaves and flowers can be used.

The extraction method of the present invention is not particularly limited, and for example, it may be extracted by heating, or may be extracted at room temperature or low temperature.
Examples of the solvent to be extracted include water, lower alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohols (1,3-butylene glycol, propylene glycol). , Glycerin, etc.), ketones (acetone, methyl ethyl ketone, etc.), acetonitrile, esters (ethyl acetate, butyl acetate, etc.), hydrocarbons (hexane, heptane, liquid paraffin, etc.), ethers (ethyl ether, tetrahydrofuran, propyl ether) Etc.). Preferred are polar solvents such as water, lower alcohols and liquid polyhydric alcohols, and particularly preferred are water, ethanol, 1,3-butylene glycol and propylene glycol. These solvents may be used alone or in combination of two or more.

  The extract may be used as it is, or may be used after concentration, dilution, filtration, decolorization with activated carbon, deodorization, or the like, if necessary. Furthermore, the extracted solution may be concentrated, dried, spray-dried, freeze-dried, etc., and used as a dried product, or it may be used after concentrating or isolating active ingredients by column purification or the like. Also good.

  In order to use the ceramide production promoter containing at least one extract selected from the group consisting of the extract of Gera and Geno shochu of the present invention for the treatment and prevention of squamous cell carcinoma, psoriasis and atopic dermatitis, it is usually systemic. Or topical or topical administration. The dose varies depending on age, body weight, symptoms, therapeutic effect, administration method, treatment time, etc., but is usually 1 mg to 5000 mg, preferably 5 mg to 1000 mg, more preferably 20 mg to 200 mg per adult. It is administered once to several times a day. Since the dosage varies depending on various conditions, an amount smaller than the above dosage range may be sufficient, or it may be necessary to administer beyond the range.

  Examples of the ceramide production promoter by oral administration of the present invention include tablets, pills, powders, granules, capsules, emulsions, solutions, suspensions, syrups, and elixirs. The above extract may be used as it is, and the excipient, extender, binder, wetting agent, disintegrant, surfactant, lubricant, dispersant, buffer, as long as the effect of the extract is not impaired. Agents, fragrances, preservatives, solubilizers, solvents and the like can be used. Specifically, lactose, sucrose, sorbit, mannitol, starch, precipitated calcium carbonate, heavy magnesium oxide, talc, calcium stearate, magnesium stearate, cellulose or derivatives thereof, amylopectin, polyvinyl alcohol, gelatin, water, Examples thereof include physiological saline, ethanol, glycerin, propylene glycol, cocoa butter, lauric fat, petrolatum, paraffin, and higher alcohol.

  Examples of the ceramide production promoter for external use of the present invention include creams, ointments, poultices and the like. The above-mentioned extract may be used as it is, and the components used for the external preparation are within the range not impairing the effect of the extract, oils, waxes, hydrocarbons, fatty acids, alcohols, esters, interfaces Components such as activators, metal soaps, pH adjusters, preservatives, fragrances, humectants, thickeners, dyes, antioxidants, chelating agents and the like can be blended.

  The amount of the extract used in the present invention is 0.0001% by weight or more, preferably 0.001 to 50% by weight, more preferably 0.001% by weight in terms of solid matter with respect to the ceramide production promoter of the present invention. A blending ratio of 01% to 10% by weight is good. If it is less than 0.0001% by weight, a sufficient effect is hardly expected. When it exceeds 50% by weight, it is uneconomical that the effect is hardly increased. The blending method may be added in advance or during the production, and may be appropriately selected in consideration of workability.

  In order to describe the present invention in detail, examples of production, formulation and experimental examples of the extract used in the present invention will be given as examples, but the present invention is not limited thereto.

Production Example 1 Geothermal hot water extract 2 L of purified water was added to 100 g of ground yellow and extracted at 95-100 ° C. for 2 hours, followed by filtration. 0.2 g was obtained.

Production Example 2 Diou ethanol extract 2 L of ethanol was added to 100 g of ground yellow and extracted at room temperature for 7 days, followed by filtration. The filtrate was concentrated to dryness to obtain 16.8 g of Diou ethanol extract.

Production Example 3 Diou 50% 1,3-butylene glycol aqueous solution extract 200 ml of purified water and 200 ml of 1,3-butylene glycol were added to 20 g of ground yellow and extracted at room temperature for 7 days, followed by filtration, 360 g of 3-butylene glycol aqueous solution extract was obtained.

Manufacture example 4 Hot water extract of Genoko pepper 1 L of purified water was added to 50 g of dried dried Gengo pepper roots and stems, extracted at 100 ° C. for 2 hours, filtered, and the filtrate was concentrated, freeze-dried, and hot 4.6 g of water extract was obtained.

Production Example 5 Gentian pepper ethanol extract Add 50 g of dried leaves and stems of ethanol to 1 g of ethanol, extract for 7 days at room temperature, filter, concentrate the filtrate to dryness, 0.6 g was obtained.

Production Example 6 50% 1,3-butyleneglycol aqueous solution extract of Genokosho 200 ml of purified water and 200 ml of 1,3-butyleneglycol were added to 20 g of dried leaves of Gennosho leaves and flowers, extracted for 7 days at room temperature, and then filtered. Thus, 330 g of a 50% 1,3-butylene glycol aqueous solution extract of Gennoshokko was obtained.

Formulation Example 1 Beverage Formulation Blending amount (g)
1. Geothermal hot water extract (Production Example 1) 0.3
2. Citric acid 0.7
3. Fructose dextrose liquid sugar 60.0
4). Fragrance 0.1
5. Purified water 38.9
[Production method] Components 1 to 4 were added to component 5, dissolved by stirring, filtered, heat sterilized, and filled into a glass bottle.

Formulation Example 2 Tablet Formulation Blending amount (g)
1. Genokosho ethanol extract (Production Example 5) 5.0
2. Corn starch 10.0
3. Purified white sugar 20.0
4). Carboxymethylcellulose calcium 10.0
5. Microcrystalline cellulose 40.0
6). Polyvinylpyrrolidone 5.0
7). Talc 10.0
[Production method] Components 1 to 5 were mixed, then an aqueous solution of component 6 was added as a binder, and granulated by a conventional method. To this, ingredient 7 was added as a lubricant and mixed, and then tableted into 100 mg tablets.

Formulation Example 3 Capsule Formulation Blending amount (g)
1. Diou ethanol extract (Production Example 2) 2.0
2. Microcrystalline cellulose 60.0
3. Corn starch 18.0
4). Lactose 18.0
5. Polyvinylpyrrolidone 2.0
[Production Method] Components 1 to 5 were mixed and granulated, and then No. 2 hard capsules were filled with 250 mg to obtain capsules.

Formulation Example 4 Cream Formulation Formulation amount (g)
1. Hot water extract of Genokosho (Production Example 4) 0.1
2. Squalane 5.5
3. Olive oil 3.0
4). Stearic acid 2.0
5. Beeswax 2.0
6). Octyldodecyl myristate 3.5
7). Polyoxyethylene cetyl ether (20E.O.) 3.0
8). Behenyl alcohol 1.5
9. Glycerol monostearate2.5
10. Fragrance 0.1
11.1,3-butylene glycol 8.5
12 Ethyl paraoxybenzoate 0.05
13. Methyl paraoxybenzoate 0.2
14 Purified water 68.05
[Manufacturing method] Components 2 to 9 are heated and dissolved and mixed, and kept at 70 ° C to obtain an oil phase. Ingredients 1 and 11 to 14 are dissolved by heating and mixed, and kept at 75 ° C. to form an aqueous phase. A water phase was added to the oil phase to emulsify, and the mixture was cooled while stirring.

Formulation Example 5 Ointment Formulation Formulation amount (g)
1. 50% 1,3-butylene glycol extract of Diou
(Production Example 3) 0.01
2. 50% 1,3-butylene glycol extract of Gennoshoco
(Production Example 6) 0.5
3. Polyoxyethylene cetyl ether (30E.O.) 2.0
4). Glycerol monostearate 10.0
5. Liquid paraffin 5.0
6). Cetanol 6.0
7). Methyl paraoxybenzoate 0.1
8). Propylene glycol 10.0
9. Purified water 66.39
[Manufacturing method] Components 3 to 6 were dissolved by heating and mixed, and kept at 70 ° C to obtain an oil phase. Ingredients 1, 2 and 7-9 were dissolved by heating and mixed, and kept at 75 ° C. to obtain an aqueous phase. An aqueous phase was added to the oil phase to emulsify, and the mixture was cooled to 30 ° C. with stirring to obtain an ointment.

Experimental Example 1 Ceramide Production Promotion Test 1 × 10 ⁵ HaCaT cells, a human epidermal keratinocyte-derived cell line, were seeded in a 60 mm dish and cultured for 4 days. Thereafter, the samples were added to final concentrations of 1, 10 and 100 μg / mL, and further cultured for 24 hours. Next, the cells were washed three times with PBS (−), collected by a rubber policeman, and lyophilized. Lipids were extracted from cells with 1 mL of chloroform: methanol (2: 1), and the amount of ceramide in the cells was determined by the fluorescence method of Kisic et al. (Kisic A. and Rapport MM, Journal of Lipid Research, 15 179-180 ( 1974)). That is, lipids were hydrolyzed with 0.15 mL of 3N hydrochloric acid at 100 ° C. for 2 hours, dried in a desiccator, removed with hydrochloric acid, and then added with 2 mL of ethyl acetate and 1.25 mL of 0.1N acetate buffer (pH 3.7). Stir well. Next, 0.25 mL of a fluorescamine solution (fluorescamine 7 mg / 25 mL acetone) was added, stirred well, then centrifuged, and the fluorescence intensity of the ethyl acetate layer was measured at Ex 410 nm and Em 490 nm. The ceramide production promotion rate was calculated by the following formula.
Ceramide production promotion rate (%) = (ceramide production amount with sample added / control ceramide production amount) × 100

  The results of these experiments are shown in Table 1. As a result, the extract of Diou and Gennosho showed an excellent ceramide production promoting effect as compared with the control.

The ceramide production promoter containing at least one extract selected from the group consisting of the present scorpion extract and genus shochu extract exhibits an excellent ceramide production promoting effect. This ceramide production promoter can treat and prevent skin diseases such as squamous cell carcinoma, psoriasis and atopic dermatitis which are highly related to ceramide by promoting the production of ceramide.

Claims (2)

A ceramide production promoter containing a hot water extract of Genokosho. A therapeutic agent for squamous cell carcinoma containing a hot water extract of Genokosho.