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Compositions and methods for lightening skin and reducing hyperpigmentation

A technology of composition and compound, applied in the field of skin brightening, pigmentation removal, and skin care, which can solve the problem of less measurement

Active Publication Date: 2018-12-21
VERSITECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Other plant extracts or polyphenols have also been tested based on the mushroom tyrosinase system, but they have been rarely assayed in cell-based systems (Solano F. et al., Pigment CellRes, 19(6) : 550-71 (2006))
[0005] Agents that reduce melanin levels by other mechanisms may be highly toxic

Method used

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  • Compositions and methods for lightening skin and reducing hyperpigmentation
  • Compositions and methods for lightening skin and reducing hyperpigmentation
  • Compositions and methods for lightening skin and reducing hyperpigmentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0209] Example 1: No apparent cytotoxicity associated with 6-CEPN and significant efficacy in reducing melanin synthesis

[0210] Materials and methods

[0211] Cell Culture and Handling

[0212] Mouse melanin A (Melan-a) cells (immortalized, non-tumorigenic mouse melanocyte cell line) were obtained from the Chinese University of Hong Kong. Cells were cultured in RPMI 1640 medium. The medium was supplemented with 10% FBS and 1% penicillin-streptomycin, and the cells were incubated at 37°C, 5% CO 2 incubate. Melanin A cells were incubated with tetradecanoylphorbol acetate (TPA) at a concentration of 200 nM. The tested chemicals (eg, 6-CEPN and kojic acid) were dissolved in 100% dimethyl sulfoxide (DMSO) at a concentration of 100 mM as stock solutions, respectively. Dilute the stock solution to the desired final concentration using culture medium immediately before use. The final DMSO concentration does not exceed 0.1%.

[0213] Assessing cell viability

[0214] Melani...

Embodiment 2

[0220] Example 2: Inhibition of tyrosinase and tyrosinase-related proteins by 6-CEPN

[0221] Materials and methods

[0222] Tyrosinase activity assay

[0223] Cellular tyrosinase activity was determined by measuring the oxidation of dopachrome in cell lysates by L-3,4-dihydroxyphenylalanine (L-DOPA). Melanin A (Melan-a) cells (1.5×10 5cells / well) were seeded in 1 mL of medium in a 24-well multiwell plate for at least 24 hours. Cells were treated with α-MSH (1 μM) alone or with α-MSH plus various concentrations of test reagents for 72 hours. Cells were washed with cold PBS and lysed in 900 μL PBS containing 1% Triton X-100. After freeze-thawing at -80°C for 30 minutes and then keeping at 25°C for 30 minutes, 100 μL of 10 mM L-DOPA was then added to each well. After incubation at 37°C for 1 hour, the absorbance of the reagents at 490 nm was measured using a Victor X4 Multi label Plate Reader (PerkinElmer, MA, USA) .

[0224] western blot

[0225] Melanin A (Melan-a) c...

Embodiment 3

[0229] Example 3: Induction of melanosome autophagy by 6-CEPN and 8-CEPQ

[0230] Materials and methods

[0231] Cell Processing for Fluorescence Microscopy

[0232] Melanin A (Melan-a) cells (1.5×10 5 cells / well) were seeded on coverslips in 6-well plates. Cells were treated with (1) α-MSH (1 μM) alone for 24 hours, (2) α-MSH (1 μM) plus rapamycin (0.5 μM) for 24 hours, (3) α-MSH (1 μM) plus 6- CEPN (10 μM), or (3) α-MSH (1 μM) plus 8-CEPQ (5 μM) were treated for 72 hours. After the cells reached a confluency level of 50%-70%, the medium was carefully removed. Cells were stained by microscope dual detection reagent (autophagy detection kit, Abcam) at 37°C for 30 minutes. Stained cells were analyzed by widefield fluorescence microscopy (6Ox magnification). Set standard FITC filter and DAPI filter to image autophagy signal and nuclear signal, respectively.

[0233] siRNA transfection and western blot analysis

[0234] Melanin A (Melan-a) cells (3×10 5 cells / well) wer...

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PUM

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Abstract

Provided is a composition. The composition comprises a compound having the structure of Formula (1). The composition reduces the synthesis of melanin by inhibiting the activity and expression levels of tyrosinase and related proteins, thereby preventing and limiting the content and distribution of melanin in mammalian skin. It also induces autophagy of melanosome, thereby restricting the synthesis, storage and transfer of melanin from melanocyte to keratinocyte. The dual modes of action allow for the efficacy of the composition at low dosage without causing toxicity to lighten skin and treat disorders associated with hyperpigmentation.

Description

field of invention [0001] The disclosed invention is generally in the field of skin care, particularly skin lightening and depigmentation. Background technique [0002] Formulations for lightening skin and / or reducing hyperpigmentation provide therapeutic and / or aesthetic benefits. Often needed to eliminate or suppress irregular pigmentation (including melasma (chloasma or localized discoloration), age spots (age spots) or chloasma (associated with sun damage or aging, sometimes manifesting as raised spots or seborrheic horns) disease) and freckles (summer freckles)). Excessive accumulation of melanin results in hyperpigmentation, which is associated with a number of diseases or conditions, including solar nevus, melasma, freckles, and post-inflammatory hyperpigmentation. In most East Asian cultures, traditional aesthetics prefer light-toned skin tones. [0003] Skin pigmentation is related to the amount and type of melanin synthesized by epidermal melanocytes and its tra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/352A61P17/00
CPCA61K45/06A61K31/352C07D311/30C07D311/32A61P17/00A61K31/353A61K2300/00A61K8/9794A61K8/9789A61K8/445A61K8/498A61Q19/02
Inventor 王明福胡舒婷吴忆箴
Owner VERSITECH LTD
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