Stimulation of the synthesis of the activity of an isoform of lysyl oxidase-like LOXL for stimulating the formation of elastic fibres

Abstract

The invention relates to the stimulation of the synthesis and of the activity of an isoform of lysyl oxidase, and more particularly of the LOXL (Iysyl oxidase-like) isoform. The invention relates notably to a method of identifying an active principle which stimulates the formation of elastic fibers. The aim of the invention is mainly to provide such a method of identification so as to provide compositions which enable stimulating the formation of elastic fibers.

What is claimed is:  
     
         1 . A composition for stimulating the formation of tissue elastic fibers, comprising the isoform of lysyl oxidase LOXL having the Sequence ID No1, or of an homologous or essentially homologous form thereof.  
     
     
         2 . A composition for stimulating the formation of elastic fibers, comprising a substance stimulating LOXL.  
     
     
         3 . The composition of  claim 2 , wherein promoting LOXL is selected from the group consisting of stimulating the activity of LOXL, stimulating the expression of a sequence of nucleotide encoding LOXL, and stimulating the expression of a sequence of peptides constituting a fraction of LOXL.  
     
     
         4 . The composition of  claim 1 , wherein the composition further comprises a second substance stimulating the expression of the protein elastin.  
     
     
         5 . The composition of  claim 4 , wherein said second substance is the substance stimulating LOXL.  
     
     
         6 . The composition of  claim 2 , wherein said substance comprises a region fixes to at least one part of the nucleotide sequence of the promoter of the human LOXL gene (Pr) (SEQ ID No3) or of an homologous or essentially homologous form thereof.  
     
     
         7 . The composition of  claim 2 , wherein said substance modulates the expression of the protein fixes to at least one part of the sequence of nucleotides of the promoter of the human LOXL gene (Pr) (SEQ ID No3).  
     
     
         8 . The composition of  claim 2 , wherein said substance is selected from the group consisting of dill, currant, cardamon, black radish, small holly, cinnamon, lactic bacteria-based fermentations, oats, potato, silk, Asea foetida gum, ethyl hexenoate and its derivatives, methyl butyrate and its derivatives, and ethyl decadienoate and its derivatives.  
     
     
         9 . A cosmetic composition, comprising an active substance as defined in  claim 1 , in admixture with a cosmetically acceptable excipient.  
     
     
         10 . A cosmetic composition, comprising an active substance as defined in  claim 2 , in admixture with a cosmetically acceptable excipient.  
     
     
         11 . A neutraceutical composition, comprising a substance as defined in  claim 1 , in admixture with an excipient acceptable for food.  
     
     
         12 . A neutraceutical composition, comprising a substance as defined in  claim 2 , in admixture with an excipient acceptable for food.  
     
     
         13 . A pharmaceutical composition, comprising a substance as defined in  claim 1 , in admixture with a pharmaceutically acceptable excipient.  
     
     
         14 . A pharmaceutical composition, comprising a substance as defined in  claim 2 , in admixture with a pharmaceutically acceptable excipient.  
     
     
         15 . A method of cosmetic care, comprising delivering to a person the composition of  claim 9 .  
     
     
         16 . A method of cosmetic care, comprising delivering to a person the composition of  claim 10 .  
     
     
         17 . The method of cosmetic care of  claim 15 , wherein the cosmetic care is selected from the group consisting of combating against the loosening of the tissues, against the loosening of the tissues during ageing, combating against the loosening of tissues during solar exposures, densifying the extracellular matrix, firming up the subcutaneous tissues, reducing skin wrinkles, slowing down the appearance of anti-wrinkles effect, improving the quality of scar tissue, improving the appearance of scars, improving the appearance and the quality of dystrophic, improving the appearance and the quality of keloid scars, and combating against stretch marks.  
     
     
         18 . The method of cosmetic care of  claim 16 , wherein the cosmetic care is selected from the group consisting of combating against the loosening of the tissues, against the loosening of the tissues during ageing, combating against the loosening of tissues during solar exposures, densifying the extracellular matrix, firming up the subcutaneous tissues, reducing skin wrinkles, slowing down the appearance of anti-wrinkles effect, improving the quality of scar tissue, improving the appearance of scars, improving the appearance and the quality of dystrophic, improving the appearance and the quality of keloid scars, and combating against stretch marks.  
     
     
         19 . A screening method for screening a substance stimulating the isoform of lysyl oxidase LOXL, wherein said method comprises the following steps: 
 placing a potentially active substance in contact with LOXL, and    analyzing the activity or the expression of LOXL, with the aim of identifying whether said potentially active substance stimulates the activity or the expression of LOXL.    
     
     
         20 . A screening method for screening a substance stimulating the formation of LOXL or of an homologous or essentially homologous form thereof, said method comprising: 
 placing a potentially active substance in contact with at least one type of cells capable of expressing the isoform LOXL of the protein lysyl oxidase (LOXL) or of an homologous or essentially homologous form thereof, and    analyzing the activity or the expression of LOXL or of an homologous or essentially homologous form thereof, with the aim of identifying whether said potentially active substance stimulates the expression of LOXL or of an homologous or essentially homologous form thereof.    
     
     
         21 . The screening method of  claim 20 , wherein said analysis of the expression of LOXL comprises an analysis selected from the group consisting of analyzing the expression of at least one nucleotide sequence encoding the protein LOXL, and analyzing the expression of a peptide sequence constituting of a peptide fraction of the protein LOXL.  
     
     
         22 . The screening method of  claim 20 , wherein said analysis of the expression of LOXL is carried out by a method selecting from the group consisting of qualitative analysis of the expression of at least one part of nucleotide sequence encoding LOXL, and quantitative analysis of the expression of at least one part of nucleotide sequence encoding LOXL.  
     
     
         23 . The screening method of  claim 20 , wherein said nucleotide sequence is the cDNA of LOXL being defined by the sequence ID No2, which is complementary to the mRNA encoding LOXL.  
     
     
         24 . The screening method of  claim 20 , wherein said analysis of the expression of LOXL uses a reverse transcription polymerase chain reaction (RT-PCR) which comprises the use of primers hybridizing with at least one part of the nucleotide sequence of the complementary DNA encoding LOXL (SEQ ID No2), in order to amplify at least one part of the nucleotide sequence encoding LOXL.  
     
     
         25 . The screening method of  claim 20 , wherein said method comprises a step of locating the expression of LOXL which is carried out on a reconstructed skin model or on a model based on biopsies, by a method selected from the group consisting of in situ hybridization of at least one part of a nucleotide sequence encoding LOXL, and immuno-detection using at least one specific antibody of LOXL.  
     
     
         26 . The screening method of  claim 20 , wherein said method comprises the comparison of the expression of LOXL with the expression of LOXL expressed in a control which does not comprise said potentially active substance.  
     
     
         27 . The screening method of  claim 20 , wherein said potentially active substance is in contact with living cells, said living cells being selected from the group consisting of fibroblasts, fibroblasts from normal human skin, fibroblasts originating from the foreskin, fibroblasts originating from a skin of an adult subject, epithelial cells, keratinocytes, keratinocytes originating from normal human skin, keratinocytes originating from the foreskin, and keratinocytes originating from a skin of an adult subject.  
     
     
         28 . The screening method of  claim 20 , wherein said potentially active substance is in contact with living cells, said living cells originate from at least one skin having a particular location, said location being the face, the abdomen, the breasts, a zone which has scars or stretch marks and said living cells being able to be characterized as being <<aged>> or as being <<exposed>> to sun's radiation or not.  
     
     
         29 . The screening method of  claim 20 , comprising the use of a model selected from the group consisting of a reconstructed skin model, a model based on biopsies, a dermis model comprising fibroblasts, and an epidermis model comprising keratinocytes, for placing in contact said potentially active substance with said cells.  
     
     
         30 . The screening method of  claim 20 , wherein said method comprises an analysis selected from the group consisting of analyzing the expression of the sequence at least of the protein elastin and analyzing the expression of a nucleotide sequence encoding the protein elastin.  
     
     
         31 . The screening method of  claim 20 , wherein said method comprises immuno-detecting the expression of the protein LOXL, with the aim of performing the traceability of neo-elastogenesis.  
     
     
         32 . The screening method of  claim 31 , wherein said immuno-detection is carried out on tissue selected from the group consisting of epithelial tissues and connective tissues.  
     
     
         33 . A method of locating the expression of LOXL in tissues with the aim of performing the traceability of neo-elastogenesis, wherein said method comprises a step of immuno-detecting the protein LOXL.  
     
     
         34 . A method of locating the expression of LOXL or of an homologous or essentially homologous form thereof in tissues with the aim of performing the traceability of neo-elastogenesis, wherein said method comprises a step of in situ hybridizing the gene encoding LOXL.  
     
     
         35 . A method of treatment of a disorder associated with the enzymatic activity of the isoform of the protein lysyl oxidase LOXL, comprising administering to a subject in need thereof a therapeutically effective amount of a composition comprising the protein lysyl oxidase LOXL or an homologous or essentially homologous form thereof.  
     
     
         36 . A method of treatment of a disorder associated with the enzymatic activity of the isoform of the protein lysyl oxidase LOXL, comprising administering to a subject in need thereof a therapeutically effective amount of a composition comprising a substance stimulating the protein lysyl oxidase LOXL, said stimulation being selected in the group consisting of stimulating the enzymatic activity of the protein lysyl oxidase LOXL, and stimulating the expression of the protein lysyl oxidase LOXL.  
     
     
         37 . The method of  claim 35 , wherein the treatment is selected from the group consisting of combating against the loosening of the tissues, densifying the extra cellular matrix, firming up the subcutaneous tissues, reducing skin wrinkles, anti-wrinkles effect, improving the quality of scar tissue, improving the quality of the appearance of scars, improving the quality of the dystrophic scars, improving the appearance of dystrophic scars, improving the quality of keloid scars, improving the appearance of keloid scars, and combating against stretch marks.