Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
  • A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
  • A61K38/46—Hydrolases (3)
  • A61K38/48—Hydrolases (3) acting on peptide bonds (3.4)
  • A61K38/4873—Cysteine endopeptidases (3.4.22), e.g. stem bromelain, papain, ficin, cathepsin H
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
  • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
  • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
  • A61P17/10—Anti-acne agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
  • A61Q19/00—Preparations for care of the skin
  • A61Q19/008—Preparations for oily skin
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
  • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
  • G01N33/6881—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids from skin
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/94—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors
  • G01N33/9446—Antibacterials
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N2500/00—Screening for compounds of potential therapeutic value
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N2800/00—Detection or diagnosis of diseases
  • G01N2800/20—Dermatological disorders

Definitions

• the present invention relates to the use of an amino acid sequence of Dermicidin, an analogue or a fragment thereof to prevent the appearance of imperfections or for the treatment and / or prevention and / or diagnosis of acne-prone or acne-prone skin, oily skin and / or aesthetic skin disorders associated therewith.
• It also relates to a cosmetic treatment method for the prevention and treatment of oily skin and / or the associated cutaneous aesthetic disorders.
• the present invention also relates to the use of a biomarker for the diagnosis of acne skin, oily skin or acne-prone skin or oily skin.
• Acne vulgaris is a multifactorial disease (face, scapular region, arms and intertriginous regions). It is the main cause of the most frequent dermatoses. It is important not to trivialize this disease and to treat it properly, because it can have disabling psychosocial consequences, in particular because of the formation of scars.
• Acne is a disease of the follicle of the sebaceous gland.
• the following pathogenic factors play a determining role in the constitution of acne:
• Retention lesions may be open or closed comedo (microcyst, micro comedo, white head). Inflammatory lesions derived from retention lesions may be papules, pustules, with indurated nodules, abscesses, fistulas, scarring.
• the clinical severity of acne was determined using the ECLA semi-quantitative rating grid.
• This grid consists of three factors: The hyperséborré or overproduction of sebum is very common and results in a skin with a bold, shiny appearance and enlarged pores. But it is also very sensitive to environmental factors such as temperature, humidity or diet. Furthermore, it has been demonstrated that Propionibacterium acnes is directly involved in the stimulation of the IGF-1 / IGF-IRcutaneous axis, which is a receptor found on the surface of the skin, and therefore induces a hyper seborrhea and an abnormal proliferation and differentiation of keratinocytes (Isard et al., 2011).
• Oily skin most commonly leads to shiny shiny skin and / or skin imperfections such as enlarged pores, thick skin texture and / or lack of blemish.
• Benzole peroxide is likely to cause skin irritation, bleaching of the hair as well as photo-allergic reactions, so it has a significant toxicity (Research in humans of toxic photo power of 5% henzoyl peroxide M Jeanmougin, J Pedreiro, J Bouchet, J Civate - Dermatology, 1983. Drug-induced photoallergic and phototoxic reactions July 2007, Vol 6, No. 4, Pages 431-443 Kevin Stein & Stephen S Scheinfeld Clindamycin, erythromycin and sodium sulphonamide in the form of salts are sensitizing substances that can cause allergies and cause resistance (D Sasseville - Progress in Dermato-Allergology Volume XVI: France 2010
• the inventors have discovered that one of the peptides derived from the cleavage of dermicidine, the antimicrobial peptide DCD-1L known to be present at the level of sweat, was active on the growth of P. acnes and could therefore be used in a cosmetic composition, in particular as an active agent for preventing and / or treating oily skin as well as the associated cutaneous aesthetic disorders.
• the subject of the invention is therefore the use of a cosmetic composition containing at least one amino acid sequence of Dermicidine, an analogue or a fragment of the latter to prevent and / or treat acne skin and / or oily skin and the associated cutaneous aesthetic disorders.
• the invention also relates to a cosmetic treatment method, in which a cosmetic composition comprising an amino acid sequence of Dermicidine, an analogue or a fragment thereof as defined below is applied to the skin.
• Another aspect of the invention is the use of Dermicidin or an amino acid sequence derived therefrom, an analog or fragment thereof, or a nucleic acid sequence encoding a DNA sequence. amino acid according to the invention, especially as a biomarker for the diagnosis of oily skin.
• the term "skin" means the whole epidemic of the human body. More particularly, the skin considered in the present invention is preferably the skin of the face, Vietnameselleté, back, and preferably the skin of the face.
• the term "acne-prone skin” means a skin presenting an acne with an ECLA score of less than 10, preferably between 1 and 9.
• acne skin within the meaning of the invention, means skin with acne with an ECLA score between 10 and 36.
• greasy skin is intended to mean a skin having a hyperseborrhoea characterized by an excessive secretion and excretion of sebum.
• a sebum level greater than 150/200 ⁇ g / cm 2 is considered to be characteristic of such oily skin.
• imperfections within the meaning of the invention is meant skin marks or temporary or permanent scars due to acne lesions that are perceived by the subject as unsightly.
• the term “prevent” means reducing the risk of occurrence or slowing down the occurrence of a given phenomenon, namely in the present invention, cutaneous marks and scars induced by pimples.
• open or closed comedon-like retention lesions (microcysts, micro comedones, white heads), inflammatory lesions derived from retention lesions such as papules, pustules, with indurated nodules, abscesses, fistulas, scarred states glossy shiny skin and / or cutaneous imperfections such as dilated pores, thick skin texture and / or flaking defect.
• the invention also relates to a cosmetic treatment method for the treatment of acne-prone skin and / or oily skin and the prevention of imperfections, in which a cosmetic composition comprising an acid sequence is applied to the skin.
• a cosmetic composition comprising an acid sequence is applied to the skin.
• the invention relates to a pharmaceutical treatment method, especially a dermatological method, in which a pharmaceutical composition comprising Dermicidin, an analogue or a fragment thereof or at least one amino acid sequence according to the invention is applied to the skin, at least one nucleic acid sequence coding for an amino acid sequence according to the invention, or at least one agent that modulates the expression or the activity, in particular the biological activity, of said amino acid sequence.
• Another aspect of the invention is the use of Dermicidin or an amino acid sequence derived therefrom, an analog or fragment thereof, or a nucleic acid sequence encoding a DNA sequence.
• amino acid according to the invention in particular as a biomarker for the diagnosis of acne-prone or acne-prone skin or oily skin.
• DCD / PIF Dermicidin / Protease Inducing Factor
• the protein precursor After proteolytic post-translational processing (mainly by cathepsin D) in sweat (Baechle, Flad et al., 2006), the protein precursor gives rise to many peptides ranging in length from 25 to 48 aa (Flad et al. Bogumil et al., 2002). Some of these peptides, such as LL37, show antimicrobial activities against S. aureus, E. coli, Enterococcus faecalis, Candida albicans and P. acnes microorganisms. The antimicrobial peptides are derived from the C-terminal portion of the protein.
• the activity of these peptides is independent of their charge (Steffen, Rieg et al., 2006) and would go through a step of interaction with bacterial phospholipids (Li, Rigby et al., 2009).
• the N-terminal part of the protein consists of a bio-logically active factor promoting cell survival (Cunningham, Hodge et al., 1998).
• the decrease in DCD observed in atopic sweat may partly explain the microbial susceptibilities of this type of skin (Rieg, Steffen et al., 2005).
• DCD has been shown to associate with bacterial phospholipids and self-organize by forming ion channels in the bacterial membrane (Paulmann, Arnold et al., 2012).
• DCD Dermicidin associated with its signal peptide is a protein of 1 10 amino acids (SEQ ID NO: 11) comprising a signal peptide of 19 amino acids (SEQ ID NO: 14) whose gene is located on chromosome 12, locus 12ql3.1. Two isoforms of this protein are listed, one shorter, isoform 1 (SEQ ID NO: 12), the other longer, isoform 2 (SEQ ID NO: 13).
• DCD Dermicidin
• Cathepsin D which takes place in sweat (Baechle, Flad et al., 2006), this protein precursor gives rise to DCD-1 (SEQ ID NO: 19) and DCD-IL (SEQ ID NO: 20) as well as many peptides ranging in length from 25 to 48 aa (Flad, Bogumil et al., 2002).
• Dermicidin is intended to mean in this application the amino acid sequences represented by SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 16, SEQ ID NO: 19 and SEQ ID NO: 20, whether or not undergoing post-translational processing.
• the term “Dermicidine” denotes the amino acid sequence represented by SEQ ID NO: 16.
• the term “Dermicidine” is intended more particularly to denote the amino acid sequence represented by SEQ ID NO: 20.
• the N-terminal part of the protein (SEQ ID NO: 15) consists of a bio-logically active factor promoting cell survival (Cunningham, Hodge et al., 1998).
• analogue of an amino acid sequence is meant any amino acid sequence having a sequence identity of at least 85%, preferably at least 90%, and more. preferably at least 95%, and even more preferably at least 99% with said sequence, and a biological activity of the same kind.
• biological activity of the same nature with respect to an amino acid sequence according to the invention is meant the antimicrobial or stimulation properties of the survival and / or cell proliferation usually attributed to Dermicidine.
• sequence identity can be determined by visual comparison or by any computer tool generally used in the field, such as BLAST programs available at www.ncbi.nlm.nih.gov and used with the default settings.
• An analogue according to the invention may be a peptidomimetic agent.
• An analogue of an amino acid sequence of the invention may result from modifications resulting from mutation or variation in the sequences of the peptides according to the invention either from the deletion or from the insertion of one or more amino acids either the substitution of one or more amino acids or alternatively splicing. Many of these changes can be combined.
• an analogue of an amino acid sequence of the invention may comprise conservative substitutions with respect to this amino acid sequence.
• hydropathic index is an index attributed to amino acids as a function of their hydrophobicity and charge (Kyte et al (1982), J. Mol Biol, 157: 105).
• amino acid sequence or an analogue thereof targeted by the present invention may be an amino acid sequence having undergone one or more post-translational processing (s).
• post-translational processing is intended to encompass all the modifications that an amino acid sequence is likely to undergo at the end of its synthesis in a cell, such as, for example, a or phosphorylation (s), thiolation (s), acetylation (s), glycosylation (s), lipidation (s), such as farnesylation or palmitoylation, rearrangement Disulfide-type and / or cleavage-type structural bonding within the peptide sequence.
• An analogue of an amino acid sequence has, moreover, substantially the same biological activity as this amino acid sequence.
• a primary amino acid sequence may comprise sites specifically recognized by protease-type enzymes, such as trypsin, which once recognition of these sites effective will induce cleavage of the sequence by proteolysis.
• protease-type enzymes such as trypsin
• the invention also extends to fragments of Dermicidin, possibly resulting from its proteolysis.
• fragment of an amino acid sequence any portion of the amino acid sequence according to the invention comprising from 3 to 48 consecutive amino acids of said sequence.
• an amino acid sequence that is suitable for the invention may be an amino acid sequence represented by a sequence chosen from SEQ ID NO: 11 to 20, in particular from SEQ ID NO: 16, SEQ ID NO : 20, an analogue, or a fragment thereof.
• an amino acid sequence that is suitable for the invention may be an amino acid sequence represented by a sequence chosen from SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, an analogue, or a fragment thereof.
• an amino acid sequence suitable for the invention may be an amino acid sequence represented by the sequence SEQ ID NO: 20, an analogue or a fragment thereof.
• a polypeptide that is suitable for the invention may also be a natural or synthetic amino acid sequence, which may optionally be obtained after enzymatic or chemical hydrolysis of Dermicidine or by chemical or biological synthesis, or by extraction from a biological tissue, such as the skin, naturally expressing this amino acid sequence or after transfection thereof, as well as the various post-translational forms thereof, or any natural or synthetic amino acids whose sequence completely or partially comprises a aforementioned amino acid sequence, for example variants and analogs.
• the present invention also relates to nucleic acid sequences SEQ ID NO: 1 to SEQ ID NO: 10 coding respectively for the amino acid sequences SEQ ID NO: 11 to SEQ ID NO: 20 of the invention. invention and their implementation in the various uses and methods according to the invention.
• nucleic acid sequence fragment means a nucleic acid sequence encoding an amino acid sequence having a biological activity of the same nature as the amino acid sequence encoded by said sequence.
• nucleic acid sequence analogue means a nucleic acid sequence having a sequence identity of at least 85%, preferably at least 90%, and more. preferably at least 95%, and even more preferably at least 99% with said sequence, and coding for an amino acid sequence having a biological activity of the same nature as the amino acid sequence encoded by said sequence.
• analog of a nucleic acid sequence is meant a nucleic acid sequence, possibly resulting from the degeneracy of the nucleic acid code, and coding for an amino acid sequence according to the invention, in particular as defined above.
• a nucleic acid sequence of the invention may be represented by at least one sequence selected from SEQ ID NO: 6 or SEQ ID NO: 10 and more preferably SEQ ID NO: 10 .
• a nucleic acid sequence of the invention may be of any possible origin, namely either animal, in particular mammalian and even more particularly human, either plant, or micro-organisms, such as for example viruses, phages, or bacteria, among others, or fungi, without prejudging the fact that they are naturally present or not in said original organism.
• the invention also relates to the isolated and purified nucleic acid sequences coding for an amino acid sequence considered according to the invention, as well as to their analogs and fragments.
• a nucleic acid sequence according to the invention may comprise a sense, antisense or interferential sequence corresponding to a sequence coding for a polypeptide according to the invention.
• the present invention also relates to cometic compositions comprising, in a cosmetically acceptable medium, an effective amount of at least one amino acid sequence of the invention.
• a composition of the invention may contain adjuvants customary in the field under consideration, such as hydrophilic or lipophilic gelling agents, preservatives, antioxidants, solvents, perfumes, fillers, filters, odor absorbers and materials. dyes, oils, water waxes and polyols.
• adjuvants customary in the field under consideration such as hydrophilic or lipophilic gelling agents, preservatives, antioxidants, solvents, perfumes, fillers, filters, odor absorbers and materials. dyes, oils, water waxes and polyols.
• compositions according to the invention are those conventionally used in the fields under consideration.
• the amount of amino acid sequence, or nucleic acid sequence of the invention, or active agent according to the invention contained in a composition of the invention also called “effective amount” is, of course depending on the nature of the asset and the effect sought and may therefore vary to a large extent.
• a composition may contain an amino acid sequence, or a nucleic acid sequence, or an active agent according to the invention in an amount representing from 0.00001% to 2.5% of the total weight of the composition, in particular in an amount representing from 0.001% to 2% of the total weight of the composition, and more particularly in an amount representing from 0.1% to 1% of the total weight of the composition.
• the term "effective amount" of a compound of the invention a sufficient and necessary amount of this compound to obtain a desired effect, and more particularly a cosmetic effect or care with regard to of the acne skin.
• the said at least one amino acid sequence represents 0.01% of the composition.
• the present invention also relates to pharmaceutical compositions, especially dermatological compositions comprising, in a physiologically acceptable medium, an effective amount of at least one amino acid sequence according to the invention, or at least one nucleic acid sequence conforming to the invention, or at least one active agent capable of modulating the activity, the expression or the maturation of said amino acid sequence or of said nucleic acid sequence.
• the term “modulating agent” or “active agent capable of modulating the expression, the processing or the activity of an amino acid sequence or nucleic acid sequence conforming to the invention "any compound capable of acting, directly or indirectly, on at least one amino acid sequence or a nucleic acid sequence in accordance with the invention, or on an element of an intracellular signaling pathway or extracellular, or a metabolic pathway, or transcriptional and / or translational regulation involving said amino acid sequence or said nucleic acid sequence.
• physiologically acceptable medium is intended to mean a medium which is suitable for the administration of a composition topically to the skin, the scalp or the lips, orally or via parenteral route such as the intradermal or subcutaneous route.
• compositions according to the invention are those conventionally used in the fields under consideration.
• the amount of amino acid sequence, or nucleic acid sequence of the invention, or active agent according to the invention contained in a composition of the invention also known as "effective amount" is, of course, a function of the nature of the asset and the effect sought and can therefore vary to a large extent.
• a composition may contain an amino acid sequence, or a nucleic acid sequence, or an active agent according to the invention in an amount representing from 0.00001% to 5% of the total weight of the composition, in particular in an amount representing from 0.001% to 5% of the total weight of the composition, preferentially from 0.1 to 5 and more particularly in an amount representing from 0.5% to 5% of the total weight of the composition and even more preferably 2.5% to 5%.
• composition at least one additional active therapeutic agent.
• the present invention relates to the cosmetic use of an effective amount of at least one amino acid or nucleic acid sequence or as an active agent for preventing and / or treating acne skin and / or oily skin and / or the appearance of the associated cutaneous aesthetic disorders.
• the present invention relates to a cosmetic, non-therapeutic method for preventing and / or treating acne-prone skin and / or oily skin and / or the appearance of associated cutaneous aesthetic disorders in an individual. in need.
• the method comprising at least one step of administering to said individual, at least one composition comprising as active agent (i) at least one amino acid sequence of the invention, in particular as defined above.
• a method or a use of the invention makes it possible to prevent and / or treat oily skin and / or the appearance of the cutaneous aesthetic disorders associated therewith.
• a method or a use of the invention makes it possible to confer and / or to find a healthy skin.
• a method of the invention may comprise the application topically to at least a portion of the skin of an individual in need thereof, in particular on the skin of the face, back and / or Vietnameselleté, scalp, at least one layer of a topical composition of the invention.
• a cosmetic process according to the invention can be implemented daily, for example for a single application per day, or a divided administration in two or three times a day, for example once in the morning and once. the evening.
• a cosmetic process according to the invention can be implemented over a period of time varying from one week to several weeks, or even several months, this period being able to be repeated after periods of non-treatment for several months, or even years. .
• a cosmetic process according to the invention can be provided for applying a cosmetic composition of the invention, for example, at a rate of 1, 2 or 3 times per day, or more, and generally on a extended duration of at least 4 weeks, or even 4 to 15 weeks, with one or more periods of interruption where applicable.
• the present invention relates to the field of biomarkers of the skin, and more particularly of oily, hyperseborrheic skin, and to their use as targets or as cosmetic active agents.
• the present invention relates to the therapeutic use of an effective amount of at least one amino acid or nucleic acid sequence of the invention or at least one agent modulating the activity, expression or maturation of said amino acid sequence or said nucleic acid sequence, and in particular a modulating agent as defined above, as active agent to prevent the appearance of imperfections and / or prevent and / or treat acne skin.
• the present invention relates to a therapeutic method, for preventing the appearance of imperfections and / or preventing and / or treating acne skin in an individual in need thereof, the method comprising at least one step of administering to said at least one composition comprising as active agent (i) at least one amino acid sequence of the invention or (ii) at least one nucleic acid sequence of the invention, or (iii) at least one less a modulating agent for the activity, expression or maturation of said sequences of the invention, in particular as defined above.
• a method or a use of the invention allows to find a healthy skin, manifested by a smooth skin without imperfections.
• a method or a use of the invention makes it possible to prevent and / or reduce the presence of a microrelief of the skin.
• a method of the invention may comprise the application topically to at least a portion of the skin of an individual in need thereof, in particular on the skin of the face, back and / or Vietnameselleté, at least one layer of a topical composition of the invention.
• a topically therapeutic method according to the invention may advantageously comprise the application of a composition of the invention, in combination simultaneously, sequentially or separately in time with an additional cosmetic or dermatological composition distinct from the composition of the invention. invention and intended for the care of the skin.
• a pharmaceutical process according to the invention may be carried out daily, for example on the basis of a single application per day, or a divided administration in two or three times a day, for example once in the morning and in the morning. times in the evening.
• a pharmaceutical process according to the invention can be implemented over a period of time ranging from one week to several weeks, or even several months, this period being able to be repeated after periods of non-treatment for several months, or even years. .
• a pharmaceutical method according to the invention may provide for an application of a composition of the invention, for example, at a rate of 1, 2 or 3 times per day, or more, and generally over a period of time. extended by at least 4 weeks, or even 4 to 15 weeks, with one or more periods of interruption if necessary.
• the present invention relates to the field of biomarkers of the skin, and more particularly to acne skin, and to their use as targets or cosmetic active agents.
• this cutaneous surface antimicrobial protein has never been classified as varying from acne and / or fat skin and even less as one of the most relevant biomarkers emerging from this type of study.
• the inventors have observed that the level of expression of Dermicidine, and more particularly peptides derived from Dermicidine and identified by the sequences SEQ ID NO: 1 1 to SEQ ID NO: 20 and in particular SEQ ID NO: 20 were systematically decreased in oily skin compared to healthy skin, especially with a ratio of 0.32 on average obtained with 3 independent experiments.
• this cutaneous surface antimicrobial protein has never been classified as varying from oily skin and even less as one of the most relevant biomarkers emerging from this type of study.
• the present invention relates to the use of at least one amino acid sequence of the invention, or at least one nucleic acid sequence of the invention, as a biomarker of a skin condition.
• the present invention relates to the use of at least one amino acid sequence of the invention, or at least one nucleic acid sequence of the invention, as a biomarker of a skin condition.
• a use according to the invention makes it possible to characterize a state of the skin such as acne and / or oily skin.
• the present invention relates to an in vitro or ex vivo method for characterizing an acne and / or fat state of the skin of a subject comprising at least the steps of:
• a) performing, in an isolated sample of a skin of a subject, a qualitative or quantitative measure of the expression, the maturation or the activity of the biomarker, and b) comparing said measurement made in step a) with a reference measurement.
• c) Conclude that a decrease in the activity, expression or maturation of said biomarker with respect to said reference measurement is indicative of an acne and / or oily or acne-prone and / or oily skin in the subject.
• biomarker means a molecule or the activity of a molecule, the presence, content or degree of activity of which is characteristic of a biological, physiological or pathological process, or the impact or effect of administering an active agent on such a process.
• Dermicidine or an amino acid sequence derived from this protein, or an analogue or a fragment thereof or a nucleic acid sequence encoding an acid sequence amine according to the invention, especially as a biomarker for the evaluation of a state of the epidermis, which may be one of the causes of "abnormal" proliferation such as P. acnes.
• a decrease in the activity, expression or maturation of said biomarker may be indicative of an acne and / or oily or acne-prone and / or oily skin.
• an increase in the activity, expression or maturation of said biomarker may be indicative of an effective cosmetic treatment to exert a beneficial effect on the skin and more preferably an effect with respect to acne and / or oily or acne-prone and / or oily skin.
• a decrease or an increase of the activity, the expression or the maturation of said biomarker can be determined by comparison with a reference measurement obtained according to any method known to man of the 'art.
• a "reference measurement” for a given parameter is a qualitative or quantitative measure of this parameter carried out under so-called “control” or "normal” conditions, for example determined in a reference sample, or determined in a sample in the absence of a treatment alleged to have an effect on the parameter.
• a reference measurement for an amino acid sequence or a nucleic acid sequence according to the invention may be a quantitative or qualitative value relating to the expression, the maturation or the activity of said sequences determined in a skin sample physio logically healthy, or determined in a skin sample, including acne skin, before a cosmetic or therapeutic treatment.
• a reference measurement is a statistical measurement, that is to say having been repeated on different samples so as to obtain an average.
• the reference measurement can be performed in parallel or sequentially with the test measurement.
• Such information may subsequently be used to determine the healthy or acne-prone and / or acne-like nature of a skin.
• the invention relates to a method, in particular in vitro or ex vivo, for characterizing a state of acneic and / or oily or acne-prone and / or oily skin.
• the qualitative or quantitative measurement of the expression, the maturation or the activity of a nucleic acid sequence of the invention can be determined by any method known to those skilled in the art.
• methods that are suitable for the invention, mention may be made of the polymerase chain reaction (PCR), quantitative (Q-PCR) or not, in the presence or absence of reverse transcriptase (RT-PCR or Q-RT-PCR), Northern-blot, ribonuclease protection assay, microarray methods, transcriptomic chip methods, oligonucleotide microarray methods, hybridization methods if you.
• agents suitable for the detection of a nucleic acid sequence of the invention and in particular of an mRNA sequence, mention may be made of labeled nucleic acid probes which can be hybridizing to a nucleic acid sequence of the invention.
• Such a nucleic acid probe can be easily obtained by any method known to those skilled in the art.
• nucleic acid sequences according to the invention can be used to produce sense and / or antisense oligonucleotide primers, which hybridize under conditions of high stringency to at least one of the sequences SEQ ID NO: 1 , SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, an analogue or fragment thereof.
• nucleic acid sequence may also be determined, indirectly, by determining the expression of the amino acid sequence encoded by said sequence, by means of any technique known in the art, such as Western blot, ELISA, BRADFORD or LOWRY method, or as indicated below.
• the qualitative or quantitative measurement, expression, maturation, or activity of an amino acid sequence of the invention may be performed by any method known to those skilled in the art.
• ELISA Enzyme Linked Immuno-Sorbent Assay
• proteomics or glycomic methods staining polypeptides in a silver dye-based polyacrylamide gel, Coomassie blue, or SYPRO
• immunofluorescence, UV absorption of immunohistochemical methods in conventional electron or confocal microscopy
• FRET fluorescence resonance energy transfer
• TR-FRET methods time resolved FRET / FRET in time resolution
• FSPIM methods fluorescence spectral imaging microscopy
• FRAP fluorescence recovery after photobleaching / fluorescence recovery after photobleaching
• reporter gene methods atomic force microscopy (AFM) methods, surface plasmon resonance methods, microcalorimetry methods, flow cytometry methods
• immunoenzymatic assay methods from more quantitative and sensitive protein solutions can in particular be used.
• These ELISA methods combine couples of capture antibodies and specific detection of the targeted antigen.
• Specifically developed commercial or polyclonal, monoclonal or recombinant antibodies can be used.
• High capacity multiplexed ELISA techniques can also be implemented. It is thus possible to mention the Luminex antibody-type multiplexed approach (for example Bioplex from Bio-Rad), of the antibody-on-flat surface type ("antibodies-arrays”) (for example, an approach proposed by MesoScale Discovery).
• an antibody may be advantageous to detect the expression of an amino acid sequence of the invention by means of an antibody, where appropriate in a labeled form.
• an antibody can be labeled with a directly detectable or detectable substance by reaction with another reagent.
• antibodies is generally meant monoclonal or polyclonal antibodies, as well as immunoglobulin fragments capable of binding an antigen and which may be produced by any genetic engineering technique known to those skilled in the art. by enzymatic or chemical cleavage of intact antibody.
• An antibody that can be used as a tool for evaluating a state of an epidemic can be obtained by any method known to those skilled in the art, as described in "Antibodies: A Laboratory Manual” , Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1990).
• iTRAQ differential proteomic method
• the term "activity" with respect to an amino acid sequence of the invention means an antimicrobial activity, an activity for stimulating cell survival, an activity for stimulating cell proliferation, or a activity of reducing or preventing aged skin and / or skin signs of aging, as indicated above.
• Such activity can be determined by any method known to those skilled in the art, such as, for example, by evaluating the proliferation or survival of epidermal cells in cultures, such as keratinocytes, or by evaluating the activity. antimicrobial on cultured bacteria, such as Staphylococcus aureus, Escherichia coli or P. acnes.
• the determination of a state of the skin or the characterization of the effectiveness of a cosmetic treatment of the skin can be carried out by measuring the variation of the expression of an amino acid sequence of the skin.
• invention and preferably represented by a sequence selected from SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19 or SEQ ID NO: 20, an analogue or a fragment thereof. this.
• the methods of the invention are particularly advantageous insofar as their implementation does not require the use of an invasive technique.
• a The epidemic sample can thus be obtained by so-called "stripping" techniques and directly analyzed by a conventional analysis technique known to those skilled in the art.
• strippings are tacky surfaces applied to the surface of the epidemic such as Blenderm ® 3M, D 'squam (commercial adhesive CuDERM), cyanoacrylate glue or the method of "stripping” varnish. Thanks to these "strippings", adherent corneocytes and the contents of their intercellular spaces can be removed and subsequently subjected to an extraction allowing access to the protein content.
• Sampling of a sample suitable for a process of the invention may also be carried out more directly by "washing" the skin surface, for example by means of turbine-type propellers such as spiral cells. as described in patent FR 2,667,778 associated with a fluid circuit, or simply by adding / removing a drop of buffer on the surface of the skin.
• one of the markers of the invention can be used for the purpose of preclinical, more effective and more rigorous selection of individuals, with a view to evaluating the efficacy of treatment or of a cosmetic active ingredient for the care of the patient. the skin.
• a biomarker of the invention may advantageously be used as indicated above to evaluate the efficacy of an active agent, in vitro, ex vivo or in vivo.
• a biomarker of the invention can be implemented to establish a personalized advice of a cosmetic treatment for an individual according to its expression profile of cutaneous biomarkers.
• the present invention relates to the use (i) of at least one amino acid sequence of the invention, or (ii) a nucleic acid sequence of the invention, for screening for active agents capable of modulating the activity, expression or processing of said amino acid sequence or said nucleic acid sequence.
• the term "expression" with respect to an amino acid sequence for example a protein or a peptide, or a nucleic acid sequence, for example an mRNA, its content or the variation of its content with respect to a reference.
• the term “maturation” with respect to an amino acid sequence for example a protein or a peptide, or a nucleic acid sequence, for example an mRNA, the modifications who follow their synthesis in a cellular environment.
• “maturation” is understood to mean post-translational modifications, such as the glycosylation or the farnesylation of certain amino acids, or the proteolytic steps leading to the elimination of sequences. so-called “signal” or “secretory” or the release of sequences with particular biological properties.
• the term “maturation” is understood to mean, for example, the alternative splicing of an mRNA.
• the term “activity” refers to a nucleic acid sequence, for example an mRNA, its translation.
• the present invention relates to the use (i) of at least one amino acid sequence of the invention, or (ii) at least one nucleic acid sequence of the invention , to characterize the effectiveness of an acne and / or oily or acne-prone and / or oily skin treatment.
• the present invention makes it possible to propose new active agents that are suitable for the prevention and / or treatment of acne and / or oily or acne-prone and / or oily skin. Screening
• the object of the present invention is to satisfy these needs.
• the present invention relates to the use of an amino acid sequence, of nucleic acids of the invention, for screening or in a screening method, in particular in vitro or ex vivo, of active agents, particularly suitable for skin care.
• the screened active agents may in particular be suitable for caring for acne-prone and / or oily skin, and / or for the prevention and / or treatment of acne, oily skin and imperfections.
• a use or a method of the invention may comprise comparing a measurement of the activity, expression or processing of an amino acid sequence or nucleic acid sequence according to the invention. invention to a reference measurement.
• a reference measurement may be as previously defined.
• a reference measurement may be a quantitative or qualitative value relating to the expression, the maturation or the activity of said sequences determined in a sample in the absence of tested active agent.
• a reference measurement can be obtained by repeating the steps of a method of the invention, and in particular steps a), b) and c) of a method of the invention as defined above, in the absence of biological or chemical compounds to test.
• the screening of an active agent capable of modulating the activity of an amino acid sequence of the invention can be done by measuring the activity or the expression of a target molecule. belonging to the signaling or metabolism pathways in which said amino acid sequence may be involved, such as, for example, a reporter gene system.
• a method of the invention can be implemented in a cell-free system, i.e. in a system not comprising cells but reproducing cellular functions, or in an isolated cell sample.
• a method according to the invention can be carried out on an isolated cell sample, an acellular sample, an isolated amino acid sequence or an isolated nucleic acid sequence of the invention, obtained by cutaneous biopsy, from cells in culture, in particular from an epidermal model, or a non-invasive skin surface sample, in particular by stratum corneum tape-stripping or by simple washing, as previously described.
• a cellular sample that is suitable for the invention, there may be mentioned a sample of keratinocytes or any other cell type of the skin expressing an amino acid sequence of the invention.
• the screening of an active agent can be carried out by measuring the variation of the expression, in the presence and in the absence of the screened active agent, of an amino acid sequence of the invention, and preferably represented by a sequence selected from SEQ ID NO: 16, SEQ ID NO: 17, or SEQ ID NO: 18, an analogue or a fragment thereof.
• an active agent of the invention may be a stimulating agent for the activity, expression or maturation of an amino acid sequence of the invention.
• modulating agent that can be screened according to a use or a process in accordance with the invention, mention may also be made of antibodies or interfering RNAs.
• the principle of the assay is based on the contact of decreasing concentrations of antimicrobial peptide (DCD-1L) with an identical P.acnes inoculum in culture medium suitable for P. acnes growth. After incubation of the 48h microplate with the P. acnes strain, the Optical Density of the sample (620 nm) is measured and the results are expressed as a percentage of calculated growth relative to a growth control without antimicrobial peptide (negative control). .
• DCD-1L antimicrobial peptide
• the minimum inhibitory concentration or MIC is the smallest concentration of antibiotic sufficient to inhibit, in the laboratory that is to say in vitro, the growth of a strain of bacteria (bacterial colony).
• the range of concentrations chosen ranged from 5 to 100 ⁇ g / mL.
• the inhibitory activity of the peptides was evaluated on the following germ: - P. acnes with TCC 6919 (Gram + bacterium)
• a cosmetic composition comprising the following ingredients (% by weight):
• a therapeutic composition comprising the following ingredients (% by weight):
• a cosmetic composition comprising the following ingredients (% by weight):

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