Method and composition for color modulation

A composition, a technology of a cosmetic composition, applied in the field of skin tone regulation

Inactive Publication Date: 2012-08-15
UNILEVER NV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Furthermore, none of the above additional information describes methods of mo

Method used

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  • Method and composition for color modulation
  • Method and composition for color modulation

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Experimental program
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Effect test

Embodiment 1

[0073] Melanosome transfer was assessed using a co-culture assay. Human epidermal melanocytes were grown to 60-80% confluence. Cells were transfected with siRNA (small interfering RNA) according to Sequitur Inc. industry-recognized and available procedures, and subsequently overlaid with HaCaT keratinocytes 72 hours later. The co-cultures were grown in phorbol ester-free keratinocyte medium (such as that available from PromoCell GmbH) and fixed 48 h after overlay to detect melanin by the Fontana-Masson staining procedure. Slides were observed using a light microscope (Carl Zeiss MicroImaging GmbH) and images were acquired using the industry-accepted AxioVision software. Ten random high power fields were analyzed per co-culture sample.

[0074] Co-culture experiments to test melanosome transfer in DDT siRNA-transfected melanocytes confirmed that melanosome transfer was reduced (inhibited) after DDT knockdown (knockdown), surprisingly showing that DDT is Plays a role in body ...

Embodiment 2

[0076] Preparation and detection of D-dopachrome substrate

[0077] In an amber 5 mL vial in H 2 Prepare a 125-mM sodium periodate working solution in O (prepared fresh on the day of the assay). Weigh 4.3 mg of dihydroxy-D-phenylalanine (D-dopa) into a 20 mL scintillation vial fitted with a pierceable septum screw cap. Using a liquid transfer cannula, first transfer 12 mL of degassed buffer into a 15 mL graduated conical tube and subsequently into a scintillation vial containing D-dopa. While stirring under nitrogen, D-dopa was dissolved for 10 minutes. The resulting working solution was 1.8 mM. Once sodium periodate is added to dissolved D-dopa, the resulting D-dopachrome is immediately used to prevent substrate degradation.

[0078] Concentrations are based on a final reaction volume of 200 microliters. Add a total of 75 µl of buffer (100 mM potassium phosphate, pH 6.8) to each well of a clear flat bottom 96-well microtiter plate. Add 25 µl of 10% glycerol / assay buffer...

Embodiment 3

[0084] Prepare a stock solution of 10 mM L-dopa (3,4-dihydroxyphenylalanine, Sigma Cat #D9628) in sodium phosphate buffer (100 mM, pH 7.0) and 0.1 mg / ml (605 units / ml ) stock solution of mushroom tyrosinase (Sigma cat #T7755) in phosphate buffered saline and stored at 4°C until use.

[0085] Test compounds (dissolved in DMSO) were first diluted in phosphate buffered saline to a working concentration of 1 mM. For each assay, 150 microliters of phosphate buffer, 10 microliters of L-dopa stock solution, and 20 microliters of each compound were added to each well of a 96-well clear bottom microtiter plate and mixed three times. Add 20 μl of mushroom tyrosinase stock solution, mix and read absorbance at 475 nm at 0, 2, 4 and 6.5 min. Plot the points as absorbance versus time and calculate the slope of the line segment. Values ​​are expressed as a percentage of the respective untreated control response. The final DMSO concentration in each sample was less than or equal to 1.0%. ...

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Abstract

The invention is directed to a method and composition suitable for skin color modulation. Particularly, the action of D-dopachrome tautomerase on melanogenesis and/or the transfer of melanin from melanosomes in keratinocytes is reduced, thereby resulting in a change in skin color.

Description

technical field [0001] The present invention relates to methods and compositions suitable for skin tone regulation, wherein it has been surprisingly discovered that D-dopachrome tautomerase is associated with skin tone. The present invention is particularly concerned with targeting D-dopachrome tautomerase and / or its receptors to surprisingly achieve skin tone regulation. The invention more particularly relates to a method of regulating skin color, wherein the method inhibits the effect of D-dopachrome tautomerase on melanogenesis and / or the transfer of melanin from melanosomes to keratinocytes. Background technique [0002] Many consumers are concerned with the characteristics of their skin. For example, consumers with age spots or freckles often prefer that these pigment spots be less noticeable. Other consumers may wish to reduce skin darkening caused by sun exposure or to lighten their natural skin tone. In order to meet the needs of consumers, many attempts have been...

Claims

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Application Information

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IPC IPC(8): A61Q19/02A61K8/35A61K8/37A61K8/44A61K8/49
Inventor J.S.巴乔尔C.A.博斯科J.L.卡德纳斯D.J.德伦南D.马雷罗S.A.罗查L.肖尔
Owner UNILEVER NV
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