Inhibitors of micro-RNAs for use for preventing and/or attenuating skin ageing

Material and Methods

[0107]Cell Culture and Transfection

[0108]Neonatal Human Primary Dermal Fibroblasts (HDFn, Cascade, Invitrogen, Carlsbad, Calif., USA) were cultured in 106 medium added with LSGS growth supplements (Cascade). Cells were passaged usually once a week, at each passage the harvested cells number and seeded cell number were recorded in order to calculate the population doublings occurring between passages and the population doubling time. At each passage different aliquots of the cells were harvested to extract in triplicate RNA and proteins and an aliquot was submitted to senescence activated β-galactosidase staining in order to assay the senescent or non-senescent state of the cells.

[0109]Human primary fibroblasts were transfected with human pre-miR 134, anti-miR-134, pre-miR-152, anti-miR-152 and scramble sequence as negative control (Ambion, Tex., USA) using the Lipofecatmine RNAimax transfection reagent (Invitrogen) according to manufacturer protocols. 24 hrs after t

Cosmetic Composition (O / W Serum)

[0134]The following composition may be prepared in a classical manner for the man skilled in the art.

[0135]The active agent is prepared as follows:

[0136]The wood of Andira coriacea (i.e. Bois d'Ange) was extracted with ethanol, yielding 386 mg of crude extract after evaporation under reduced pressure. The crude extract was then purified by reverse phase preparative HPLC(C18 Varian Pursuit XRS, 250 mm×41.1 mm×5 nm) using a gradient of H2O+0.1% formic acid and MeOH (from H2O+FA / MeOH 70:30 to 60:40 in 17 min at 60 mL min-1) to afford pure Coatline A (12.3 mg) and Coatline B (40.2 mg).

[0137]

INCI name% (w / w)WaterQSP 100.00Chelating agent0.05pH balance0.05Preservatives0.05Glycol3.25AMMONIUM ACRYLOYLDIMETHYLTAURATE / VP1.20COPOLYMERACRYLATES / C10-30 ALKYL ACRYLATE0.20CROSSPOLYMERGLYCERIN3.00GLYCERYLPOLYMETHACRYLATE4.18SODIUM ACETYLATED HYALURONATE0.05Oil10.00 ALCOHOL8.00PERFUMES0.30Coatlines obtained as described above0.05