Stimulation of the synthesis of the activity of an isoform of lysl 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 (lysyl 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.

1 . A composition for stimulating the formation of tissue elastic fibers, comprising the isoform of lysyl oxidase LOXL having the amino acid sequence of SEQ ID NO: 1, 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 the stimulating of 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 the second substance is the substance stimulating LOXL. 
     
     
         6 . The composition of  claim 2 , wherein the substance comprises a region which fixes to a part of the nucleotide sequence of the promoter of the human LOXL gene (SEQ ID NO: 3) or of an homologous or essentially homologous form thereof. 
     
     
         7 . The composition of  claim 2 , wherein the substance modulates the expression of the protein which fixes to a part of the sequence of nucleotides of the promoter of the human LOXL gene (SEQ ID NO: 3). 
     
     
         8 . The composition of  claim 2 , wherein the 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 the composition of  claim 1 , in admixture with a cosmetically acceptable excipient. 
     
     
         10 . A cosmetic composition, comprising the composition of  claim 2 , in admixture with a cosmetically acceptable excipient. 
     
     
         11 . A neutraceutical composition, comprising the composition of  claim 1 , in admixture with an excipient acceptable for food. 
     
     
         12 . A neutraceutical composition, comprising the composition of  claim 2 , in admixture with an excipient acceptable for food. 
     
     
         13 . A pharmaceutical composition, comprising the composition of  claim 1 , in admixture with a pharmaceutically acceptable excipient. 
     
     
         14 . A pharmaceutical composition, comprising the composition of  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 for a substance stimulating the isoform of lysyl oxidase LOXL, wherein the method comprises the following steps: placing a potentially active substance in contact with LOXL, and analyzing the activity of LOXL, with the aim of identifying whether the potentially active substance stimulates the activity of LOXL. 
     
     
         20 . A screening method for screening a substance stimulating the expression of LOXL or of an homologous or essentially homologous form thereof, the method comprising: placing a potentially active substance in contact with 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 the 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 the 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 the 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 21 , wherein the nucleotide sequence is the cDNA of LOXL being defined by the sequence ID No 2, which is complementary to the mRNA encoding LOXL. 
     
     
         24 . The screening method of  claim 21 , wherein the 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 NO: 2), to amplify at least one part of the nucleotide sequence encoding LOXL. 
     
     
         25 . The screening method of  claim 20 , wherein the 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 in situ hybridization of at least one part of a nucleotide sequence encoding LOXL, and immuno-detection using at least one specific antibody to LOXL. 
     
     
         26 . The screening method of  claim 20 , wherein the method comprises comparing the expression of LOXL with the expression of LOXL expressed in a control which does not comprise the potentially active substance. 
     
     
         27 . The screening method of  claim 20 , wherein the potentially active substance is in contact with living cells, the living cells 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 the potentially active substance is in contact with living cells, the living cells originating from at least one skin having a particular location, the location being the face, the abdomen, the breasts, a zone which has scars or stretch marks and the living cells being able to be characterized as aged or as exposed to sun's radiation or not. 
     
     
         29 . The screening method of  claim 28 , 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 the potentially active substance with the 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 the immuno-detection is carried out on tissue selected from the group consisting of epithelial tissues and connective tissues. 
     
     
         33 - 34 . (canceled) 
     
     
         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, the stimulation being selected from 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. 
     
     
         38 . A method of stimulating the formation of skin elastic fibers comprising applying to the skin an effective amount of a substance which stimulates the enzymatic activity or expression of the lysyl oxidase isoform LOXL. 
     
     
         39 . The method of  claim 38 , wherein the lysyl oxidase isoform LOXL has the amino acid sequence of SEQ ID NO: 1. 
     
     
         40 . The method of  claim 38 , wherein the lysyl oxidase isoform LOXL stimulates the expression of the protein elastin. 
     
     
         41 . The method of  claim 38 , wherein the substance comprises a region which fixes to at least one part of the sequence of nucleotides of the lysyl oxidase isoform LOXL promoter of SEQ ID NO: 3. 
     
     
         42 . The method of  claim 38 , wherein the substance modulates the expression of a protein which fixes to a part of the sequence of nucleotides encoding the lysyl oxidase isoform LOXL promoter of SEQ ID NO: 3. 
     
     
         43 . The method of  claim 38 , wherein the substance is selected from the group consisting of an extract of dill, an extract of cardamon, an extract of currant, an extract of black radish, an extract of small holly, an extract of cinnamon, a lactic bacteria-based fermentation, an extract of oats, an extract of potato, a silk protein,  Asea foetida  gum, ethyl hexenoate, methyl butyrate, ethyl decadienoate, and combinations thereof. 
     
     
         44 . The method of  claim 43 , wherein the effective amount of the substance is between 0.1% and 5% (v/v). 
     
     
         45 . The method of  claim 38 , wherein the stimulation of the skin elastic fibers provides a cosmetic benefit to the skin. 
     
     
         46 . The method of  claim 45 , wherein the cosmetic benefit is combating against the loosening of tissues, for densifying the extracellular matrix, for firming up the subcutaneous tissues, for reducing skin wrinkles, for exerting an anti-wrinkle effect, for improving the appearance of scars, or for combating against stretch marks.