Hairless protein-interacting partner complexes and methods thereof for the beautification and/or improvement of mammalian skin

A technology of mammals and compounds, applied in chemical instruments and methods, compound screening, drug combination, etc., can solve problems such as unidentified interactions

Inactive Publication Date: 2006-12-20
PROCTER & GAMBLE CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

While those skilled in the art have identified some interactions between HR and thyroid hormone receptors (TRs), the above interactions between HR and other proteins in keratinocytes of mammalian skin have not been identified

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0094] Example 1 - Interaction partners (IP's) of hairless proteins

[0095] This example uses the yeast two-hybrid technology to provide a hairless protein (HR) interacting partner (HR-IP). The HR cDNA corresponding to the C-terminal portion of the HR protein (amino acid residues 490 to 1182, hereinafter referred to as HRt) was cloned into a two-hybrid bait vector for expression of Lex A DNA BD-HRt in yeast. Yeast cells expressing BD-HRt were transformed with a human keratinocyte cDNA library established with an activation domain (AD) vector. Synthetic interaction partners provided by the present invention are listed in Table 1 and include molecules involved in cell cycle, cell differentiation, transcription, nuclear transport, signal transduction, cell integrity and mitochondrial apparatus. Thus, HRt (and HR) behave as well-connected, multifunctional proteins.

[0096] Material

[0097] The mouse early anagen cDNA library was constructed by Procter & Gamble staff using st...

Embodiment 2

[0111] Example 2 - Screening for compounds that disrupt the BD-HRt>

[0112] AH109 was used for compound screening. AH109 (a component of the Matchmaker Two-Hybrid 3 (Cat. No. K1612-1) BD Biosciences Clontech, Palo Alto, CA) is a suitable Y2H interacting yeast reporter strain. It contains ADE2, HIS3, lacZ and MEL1 reporter genes, each using a different GAL4-responsive promoter. Various Y2H recombinant plasmids were transformed into yeast strain AH109 using a lithium acetate-mediated yeast transformation protocol provided by the supplier of the Y2H system (BD Biosciences Clontech, Palo Alto, CA). Transformed cells of the combined AD and BD plasmids were selected based on their ability to grow on Leu and Trp double depleted medium (DDO).

[0113] The interaction between the AD and BD fusion proteins (Y2H interaction) will bring the AD and BD domains of the GAL 4 protein closer to each other, thereby reconstituting a functional GAL 4 protein. ...

Embodiment 3

[0115] Example 3 - Screening for compounds that enhance or disrupt the BD-HRt>

[0116] Increase or decrease in α-galactosidase activity of AH109 (BD-HRt>

[0117] Microplates with TDO medium were seeded with freshly cultured yeast cells with an optical density of 0.05 at 650 nm (OD650), treated with various test compounds at 100 to 400 uM or with vehicle control (DMSO) the microplate. Stock solutions of test compounds at 10 to 20 mM were prepared in DMSO at final concentrations of 100 to 400 [mu]M when assayed. The final test mixture containing cells, TDO and compound (or DMSO alone) was adjusted to 100 ul and the amount of DMSO was kept at or below 2%. Plates were incubated at 25°C for 3 to 4 days without shaking. Plates were briefly agitated to obtain a homogeneous cell suspension prior to addition of assay buffer. Reporter gene enzyme (α-galactosidase) activity was...

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PUM

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Abstract

Hairless protein interacting partner complexes, agonist and / or antagonist compounds, compositions and products comprising same and methods thereof. Hairless protein is indispensable for the beautification and / or improvement of mammalian skin. Therefore, such interacting complexes are useful for screening test compounds for the beautification and / or improvement of mammalian skin.

Description

Field of Invention [0001] The present invention relates to hairless protein interacting partner complexes, antagonist or agonist active compounds, compositions and products comprising the same, and methods thereof. The complexes can be used to screen detection agents and identify compounds that confer substantial beautification and improvement benefits to mammalian, particularly human, skin. Background of the Invention [0002] People have always been obsessed with the pursuit of appearance, especially face. But the pursuit of skin and body appearance is also growing. In fact, many people believe that there is an intrinsic link between physical appearance and self-esteem, mate choice, career advancement, and general social acceptance. The condition of human skin typically peaks at the age of 20 and later thins and loses elasticity. Not surprisingly, U.S. consumers spent about $2.8 billion in 2001 on products that claim to slow or reverse skin aging. The demand for improv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50C07K14/47
CPCG01N2500/02C07K14/4702C07K14/47A61P17/16
Inventor K·斯里科里斯纳R·A·勒伯夫
Owner PROCTER & GAMBLE CO
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