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

Inactive Publication Date: 2004-12-16
CENT NAT DE LA RECHERCHE SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

0090] Advantageously, this method of treatment enables performing a treatment selected from combating against the loosening of the tissues, notably when the loosening of the tissues is observed during ageing or during solar exposures, densifying the extracellular matrix, firming up the subcutaneous tissues, reducing skin wrinkles, anti-wrinkles effects, improving the quality of scar tissue and the app

Problems solved by technology

Similarly, in scars, the network of elastic fibers does not form correctly.
The prior art does not enable providing criteria which enable evaluating the impact of an active principle in dermato-cosmetology on a functional neo-elastogenesis.
In this context, it is also very difficult to obtain objective criteria enabling the impact of these actives to be judged.
Furthermore, at the present time, animal experimentation is forbidden in cosmetics in Europe and human experimentation is ethically disputed.
It is therefore unacceptable to the inventors to carry out a screening metho

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0135] The invention has first of all covered the development of novel specific antibodies of LOX and LOXL but which are capable of detecting their mature forms. The antibodies were developed against the mature regions of LOX and LOXL. The antigenic regions were selected in order to present the minimum of similarity with the corresponding regions on the other isoforms of the lysyl oxidases (LOs). The antibodies obtained against the regions of the peptides LOX.sup.V228-S280 were called anti-LOXmat and similarly for the antibodies obtained against the region of the peptides LOXL.sup.R231-G368 called anti-LOXLpro.

[0136] In FIG. 1: description of the sequences of the LO defined for giving the specific antibodies: This figure represents the steps which have led to the selection of the antigenic regions in order to develop the anti-LOX and anti-LOXL antibodies.

[0137] FIG. 1(A): Schematic representation of hLOX (human LOX protein) and hLOXL (human LOXL protein).

[0138] The sequences of hLOX...

example 2

Immuno-Detection of LOX and LOXL of Muscle Cells by Virtue of the Novel Antibodies Anti LOX and Anti-LOXL

[0151] On FIG. 2:

[0152] FIG. 2 represents photographs of electrophoreses which were carried out as indicated below. These electrophoreses demonstrate the characterization of the mature proteins of LOX and LOXL, of smooth muscle cells (SMC) by virtue of the antibodies anti-LOX and anti-LOXL, identified in Example 1.

[0153] The proteins of the cell strain (L) and of the cell culture medium (M) of a cell line of rat smooth muscle (developed by Jean-Marie Daniel Lamaziere, Bordeaux) were extracted and detected by western blotting by using the antibodies anti-LOXLmat, anti-LOXmat, anti-LOXLpro and anti-LOXpro. The cells were cultivated at 37.degree. C. in an atmosphere of 5% CO.sub.2 in DMEM medium (Sigma) containing 10% foetal calf serum, 2 mM glutamine and 50 .mu.g / ml gentamycin.

[0154] The cell strain proteins, which are washed twice with PBS buffer, were extracted for 2 hours at 4.d...

example 3

Demonstration of the Role of LOX and LOXL in Elastogenesis

[0158] The inventors have demonstrated that the LOX and LOXL proteins can be associated with the formation of connective tissue in the dermis of reconstructed skin models by immuno-histochemistry (FIG. 3). This demonstration was obtained without any ambiguity by virtue of the use of anti-LOX and anti-LOXL antibody couples, directed against the pro-enzymatic regions and mature regions of the two enzymes (LOX and LOXL). On FIG. 3 representation is made of the immuno-histological detection of LOXL and LOX in the reconstructed skin (RS) and normal human skin.

[0159] The immuno-detection of LOXL (A, C, E, G) on the reconstructed skin at days 16 (A), 35 (C), and 45 (E), by using anti-LOXL.sup.R231-G368 (A, C, E) or anti-LOXL.sup.R231-G368 adsorbed with the corresponding peptide GST-LOXL.sup.R231-G368 before the immuno-detection (G). The immuno-detection of LOX (B, D, F, H) at days 16 (B), 35 (D), and 45 (F), by using anti-LOX.sup.V2...

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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.

Description

[0001] The invention relates to the stimulation of the activity of an isoform of lysyl oxidase, and more particularly of the LOXL (lysyl oxidase-like) isoform.STATE OF THE ART[0002] The properties of resistance and of elasticity of the skin and of the mucous membranes are essentially defined by the collagen fibers and elastin fibers of the dermis. Elastin is a protein which makes up the elastic fibers by combining itself with other molecules such as fibrillins and MAGPs (Microfibrillar Associated Glycoproteins).[0003] The elastic fibers are formed of elastin deposited on the microfibrils. Elastin is synthesized in the form of soluble tropoelastin which acquires its physico-chemical properties (insolubility, elasticity) after the intra- and inter-molecular cross-linking of it by a lysyl oxidase, and its deposit on the microfibrils. The elastic fibers are responsible for the elastic property of the organs which contain them (vessels, pulmonary parenchyma, elastic cartilages, skin . . ...

Claims

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Application Information

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IPC IPC(8): A23L1/30A61K8/66A61K31/22A61K31/23A61K35/74A61K36/185A61K36/23A61K36/31A61K36/54A61K36/81A61K36/896A61K36/899A61K36/9064A61K38/43A61K38/44A61P17/00A61Q19/00A61Q19/08C12N9/06C12Q1/26C12Q1/68G01N33/68
CPCA61K8/66C12Y104/03013A61K36/185A61K36/23A61K36/31A61K36/54A61K36/81A61K36/896A61K36/899A61K36/9064A61Q19/00A61Q19/08C12N9/0022G01N2500/04A61K35/74A61K38/00A61K2300/00A61K38/44A61P17/00A61P17/02A61K8/97
Inventor PERRIER, ERICCENIZO, VALERIEBOUEZ, CHARBELSOMMER, PASCALDAMOUR, ODILEGLEYZAL, CLAUDINEANDRE, VALERIEREYMERMIER, CORINNE
Owner CENT NAT DE LA RECHERCHE SCI
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