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Drugs for ameliorating itch, rough skin or hypersensitive skin or for whitening via inhibition of the production and release of stem cell

A technology of stem cell factor and skin roughness, which is applied in the field of drugs for improving itching, rough skin or skin sensitivity or for whitening by inhibiting the production and release of stem cell factor, which can solve the problems of high cost and time-consuming

Inactive Publication Date: 2005-04-27
SHISEIDO CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the high cost and time-consuming of such animal experiments, it has not been a suitable method for screening a wide variety of botanical preparations and drugs, so the development of active ingredients and plants that can conveniently and economically screen for the activity of inhibiting the production and release of SCF is required. Method of preparation

Method used

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  • Drugs for ameliorating itch, rough skin or hypersensitive skin or for whitening via inhibition of the production and release of stem cell
  • Drugs for ameliorating itch, rough skin or hypersensitive skin or for whitening via inhibition of the production and release of stem cell
  • Drugs for ameliorating itch, rough skin or hypersensitive skin or for whitening via inhibition of the production and release of stem cell

Examples

Experimental program
Comparison scheme
Effect test

experiment Embodiment 1

[0064] Analysis of SCF released by desiccation-stimulated keratinocytes and screening for inhibitors

[0065] Commercially available human epidermal keratinocytes (Newborn, CryoNHEK-Neo, Sanko Junyaku Co., Ltd.) were cultured using a commercially available serum-free medium (Defined Keratinocyte-SFM, Gibco Industries, Inc.). cells at 1 x 10 5 cells / mL were plated in a 12-well microplate and incubated at about 37°C for about 72 hours to confluence.

[0066] Various plant extracts shown in Figures 1 and 2 extracted with water or ethanol were dissolved in 70% ethanol to 2% w / v.

[0067] Each test plant extract was added to the culture solution at a final concentration of 0.005% w / v, and cultured at 37°C for 24 hours. A culture containing only ethanol (EtOH) was also grown as a control. To check cell viability, i.e. to determine the cytotoxic effect of the plant extracts tested, alamarBlue TM (Biosource International) was added at 10%, and the fluorescence intensity (excitatio...

experiment Embodiment 2

[0074] Accelerated expression of SCF on keratinocyte membrane induced by different stimuli

[0075] After 1.5 million keratinocytes were placed on a 10 cm plate and cultured for 72 hours, the cells were stimulated by one of the following methods: (1) Replace the medium with PBS(-), and use UVB at 20mJ / cm 2 Irradiated cultures were immediately followed by PBS (-) replaced with medium, (2) addition of forskolin, or (3) addition of theophylline. After culturing for 24 hours, the cells were collected and dispersed in 200 μl of 50 mM phosphate buffer (pH 7.8) + protease inhibitor solution, and then disrupted by an ultrasonic disruptor at 4° C. for 5 times for 30 seconds each. After centrifugation at 10,000 g for 20 minutes at 4°C, the supernatant was further centrifuged at 100,000 g for 60 minutes at 4°C. The resulting precipitate was dissolved in 100 μl of 25 mM phosphate buffer (pH 6.8) + 0.1% Triton-X100 solution to obtain a protein extract of the membrane fraction. The protei...

experiment Embodiment 3

[0078] Analysis of SCF expressed on UV-stimulated keratinocyte membranes and screening for inhibitors

[0079] After 1.5 million keratinocytes were placed on a 10cm plate and cultured for 72 hours, the medium was replaced with PBS(-), and UVB was used at 20mJ / cm 2 Cultures were irradiated and PBS(-) was immediately replaced with medium. After adding the plant preparation to be screened and incubating for 24 hours, the cells were collected, then dispersed in 200 μl of 50 mM phosphate buffer (pH 7.8) + protease inhibitor solution, and the cells were disrupted with an ultrasonic disruptor at 4 °C5 times, 30 seconds each time. After centrifugation at 10,000 g for 20 min at 4°C, the supernatant was further centrifuged at 100,000 g for 60 min at 4°C. The resulting precipitate was dissolved in 100 μl of 25 mM phosphate buffer (pH 6.8) + 0.1% Triton-X100 solution to obtain a protein extract of the membrane fraction. The protein content of the solution was measured by well-establish...

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Abstract

The invention provides a screening method for active ingredients which exhibit effects of ameliorating pruritus, rough skin or sensitive skin, or effects of skin whitening, by inhibiting production and / or release of stem cell factor (SCF), as well as medicaments for pruritus, rough skin, sensitive skin and / or skin whitening which contain the active ingredients.

Description

technical field [0001] The present invention relates to a screening method for an active ingredient that exhibits an effect of improving itching, rough skin or skin sensitivity or an effect of skin whitening by inhibiting the production and / or release of stem cell factor (hereinafter referred to as "SCF"), and Concerning medicines containing these active ingredients for itching, rough skin, sensitive skin and / or skin lightening. Background technique [0002] Various therapeutic agents, skin external preparations, cosmetics and the like are generally used for improving itching, rough skin or skin sensitivity or for whitening the skin. As active ingredients of commonly used drugs or cosmetics that have the effect of improving itching, rough skin, or skin sensitivity, anti-inflammatory agents or amino acids, polysaccharides, lipids, etc., and various animal or plant extracts that have anti-inflammatory effects or high hygroscopic effects have been used because of their excelle...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K8/00A61K8/96A61K8/97A61K36/00A61K36/05A61K36/18A61K36/28A61K36/48A61K36/60A61K36/73A61K36/82A61K36/87A61P17/00A61P17/04A61P17/16A61Q19/00A61Q19/02G01N33/15G01N33/50G01N33/566G01N33/68
CPCG01N2500/10G01N33/502G01N2800/20A61Q19/00G01N33/5044G01N33/6881A61Q19/005G01N33/5008A61K8/97A61Q19/02G01N33/5023G01N2333/475A61K8/9722A61K8/9767A61K8/9789A61K8/9794A61P17/00A61P17/04A61P17/16
Inventor 芦田丰青木宏文藤原留美子
Owner SHISEIDO CO LTD
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