saspase-flg2 complex, modulators and uses thereof

A use, technology of compounds, applied in the field of cosmetology, prophylaxis and/or treatment of aesthetic defects or pathological disorders of the skin and/or its coverings, therapeutic or dermatological care, achieving good efficacy, well tolerated results

Inactive Publication Date: 2017-03-29
LOREAL SA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Many aspects of the mechanisms underlying the regulation of the balance between proliferation and differentiation of epidermal cells remain to be determined

Method used

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  • saspase-flg2 complex, modulators and uses thereof
  • saspase-flg2 complex, modulators and uses thereof
  • saspase-flg2 complex, modulators and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0339] Enzyme activation of SASPase caused by FLG2

[0340] a- Materials and Method

[0341] Two forms of recombinant filaggrin-2 (FLG2) were prepared in the parent plasmid pET32c (Novagen) or pEB6 (HBGX), respectively corresponding to Q5D862 (Uniprot / Swissprot ref.) fused to thiol at its N-terminus The amino acid sequence 2-213 (active form) and the amino acid sequence 81-212 (inactive form) of the affinity tag of reductin-His-S.Tag (THX-His-S-tag) (FLG2 2-213 -S-His-THX and FLG2 81-212 -S-His-THX).

[0342] These recombinant chimeric proteins are hereinafter referred to as FLG2-Hs=FLG2 2-213 -S-His-THX (active form) and FLG2-AbD=FLG2 81-212 -S-His-THX (inactive form) said.

[0343] The 28-kDa recombinant SASPase corresponding to the amino acid sequence 85 to 343 of the sequence Q53RT3 (Uniprot / Swissprot ref.) was in accordance with Bernard et al. (J. Invest. Dermatol., 2005, 125:278-287) (SASPase 85-343 ) Is prepared as described.

[0344] Dissolve SASPase in PBS buffer, and in the...

Embodiment 2

[0353] Screening of reagents regulating SASPase-FLG2 interaction

[0354] The SASPase-FLG2 interaction, and thus the stimulation of SASPase proteolytic activity, is used in a method for screening active compounds capable of modulating the interaction.

[0355] The established method is based on the FLG2-SASPase interaction and uses homogeneous time-resolved fluorescence (or HTRF) technology.

[0356] Recombinant SASPase protein (containing the amino acid sequence 85-343 of Q53RT3), mutated at its active site (D212A of Q53RT3), and at its C-terminus, and GST (from Schistosoma japonicum) glutathione S transferase ) The affinity tag is fused and prepared in the vector pGEX-4-T3 (GenBank U13855) (SASPase 85-343D212A -GST). The D212A mutation was introduced to inactivate SASPase and promote stable interaction with FLG2.

[0357] The recombinant FLG2 protein (including the amino acid sequence 2-213 of Q5D862) fused to the thioredoxin-His-S.Tag affinity tag at its N-terminus was prepared ...

Embodiment 3

[0388] The effect of compound "M7" on the differentiation and / or proliferation of epidermal cells should

[0389] a- Materials and Method

[0390] The reconstructed human epidermis Episkin J6 (No. 09-EPIS-015) model was immersed in an oven inserted at 37°C (containing 5% CO 2 ) Is cultured in 3.5ml Episkin differentiation medium, and the medium is changed every 2 days.

[0391] by using The provided differentiation medium (including the test concentration of the compound or the buffer solution (0.1% final concentration of DMSO) as a control) culture the epidermal model, and test the compound "M7" at the concentration of 10 μM, 1 μM, and 0.5 μM.

[0392] Evaluate compounds by measuring the thickness of the stratum corneum and measuring the expression and / or activation of transglutaminase I (TGI), caspase 14, SASPase, corneal chain protein, desmoside glycoprotein 1, or protein Ki67 The effect of "M7" on the differentiation and / or proliferation of epidermal cells.

[0393] The thickne...

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Abstract

The present invention relates to compounds suitable for modulating the interaction between a first partner protein and a second partner protein, respectively natural Aspartic acid protease SASPase and filaggrin-2 or FLG2 as compounds for the treatment and / or prevention of aesthetic defects of the skin and / or its covering associated with an imbalance in the differentiation and / or proliferation of epidermal cells active agent.

Description

Technical field [0001] The present invention relates to a new type of cosmetic target or treatment target useful for skin and its covering, and also relates to its use for the purpose of beauty, treatment or dermatological care. [0002] More precisely, the present invention relates to novel protein complexes involved in the differentiation and / or proliferation of epidermal cells, their use as tools for screening novel cosmetic or therapeutic active agents, the identified novel active agents, and the novel activities The agent is used to prevent and / or treat aesthetic defects or pathological disorders of the skin and / or its covering. Background technique [0003] The term "skin" refers to the entire skin of the body, including the scalp and mucous membranes. [0004] The term "skin covering" refers to body hair, eyelashes, hair and nails. [0005] The skin or epidermis is usually divided into a basal layer of keratinocytes that constitute the germinal layer of the epidermis, a spinou...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K8/49A61Q19/00
CPCA61K8/4953A61K31/519A61K2800/78A61Q15/00A61Q19/007A61Q19/08A61P17/00G01N33/6845G01N2500/02A61K8/30A61K8/49A61Q19/00G01N33/6881
Inventor D·伯纳德A·托马斯-克里基努恩R·鲁佐特B·马勒
Owner LOREAL SA
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