Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
  • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
  • C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
  • C12Q1/32—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase involving dehydrogenase
• • 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
  • A61K8/66—Enzymes
• • 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
• • 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
  • A61P17/08—Antiseborrheics
• • 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
• • 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
  • G01N2800/202—Dermatitis

Definitions

• the subject of the present invention is the use of an enzyme with dehydrogenase-reductase activity as a research tool in the disorders associated with hyperseborrhoea, in particular acne.
• the subject of the invention is also the use of this enzyme for the identification of a compound intended for the treatment of acne, seborrheic dermatitis and / or skin disorders associated with hyperseborrhoea, as well as a method for treating identification of such a compound.
• Acne is a multifactorial disease characterized by abnormal production of sebum and ductal coming, followed by bacterial colonization and inflammation.
• Today, the most effective product to treat this pathology is a synthetic retinoid, 13-cis retinoic acid or isotretinoin (Roaccutane TM). This product, however, has severe side effects, including teratogenicity.
• 13-cis retinoic acid is effective against oral acne ("Diagnosis and Treatment of Acne", Feldman et al., Am Fam Physician, 2004 May 1; 69 (9): 2123-30), its mechanism of action is still unknown.
• the main advantage of 13-cis retinoic acid is its ability to significantly decrease sebum production ("Effect of oral 13-cis retinoic acid at three dose levels on sustainable secretions of sebum secretion and on acne", Stewart ME and al, J Am Acad Dermatol, 1983 Apr; 8 (4): 532-8 and "Isotretinoin - an explanation for the long-term benefit", Cunliffe WJ et al, Dermatologica, 1987; 175 Suppl 1: 133-7).
• 13-cis retinoic acid is much more effective orally than other natural or synthetic retinoids (Cunliffe WJ, Norris JF "Isotretinoin-an explanation for long-term benefit", Dermatologica, 1987; 175 Suppl 1: 133-7).
• Original pharmacokinetic properties have been proposed to explain this efficacy.
• RAR Retinoic Acid Receptors
• a metabolite derived from dihydroxytestosterone, 3 ⁇ -androstanediol glucuronide Effect of oral isotretinoin treatment on skin androgen receptor levels in acneic patients", Boudou P et al, J Clin Endocrinol Metab, 1995 Apr; 80 (4): 1158-61 and "Isotretinoin, tetracycline and circulating hormones in acne, Palatsi R et al., Acta Derm Venerol, 1997 Sep; 77 (5): 394-6).
• Roaccutane may act by altering the intracellular androgenic concentration through the inhibition of enzymes involved in the most effective androgen catabolism, DHT (Expression cloning and characterization of oxidative 17beta and 3alpha- hydroxysteroid dehydrogenases from rat and human prostate ", Biswas MG et al., J Biol Chem, 1997 Jun 20; 272 (25): 15959-66 and” Cloning of the human RoDH-related short chain dehydrogenase gene and analysis of structure " , Kedishvili NY et al., Chem Biol Interact, 2001 Jan 30; 130-132 (1-3): 457-67).
• RoDHs have been identified in humans. These enzymes are cytosolic or microsomal, work with NAD or NADP as a cofactor, and are sensitive to different retinoids. Their mechanism of action is multifunctional, since they may have some activity retinol dehydrogenase, but also 3 ⁇ -hydroxy or 17 ⁇ -hydroxy steroid dehydrogenase.
• these enzymes include RODH or HSE (RoDH-like 3 ⁇ -HSD or 3-hydroxysteroid epimerase) ("Expression cloning and characterization of oxidative 17beta and 3alpha-hydroxysteroid dehydrogenases from rat and human prostate", Biswas MG and al, J Biol Chem, 1997 Jun 20; 272 (25): 15959-66 and "Cloning of the human RoDH-related short chain dehydrogenase gene and analysis of structure", Kedishvili NY et al., Chem Biol Interact, Jan 2001 130-132 (1-3): 457-67), RoDH-4 or Microsomal NAD + -dependent retinol dehydrogenase 4 ("cDNA cloning and characterization of a new human microsomal NAD + - dependent dehydrogenase that oxidizes all-trans-retinol and 3alpha-hydroxysteroids ", Gough WH et al, J Biol Chem, 1998 Ju 31, 273
• DHRS9 called RODH16 in mice
• RAR receptors (o ⁇ ) and ALDH1 RAR receptors (o ⁇ ) and ALDH1
• o ⁇ RAR receptors
• ALDH1 ALDH1
• It is present mainly in the anagenic phases of the hair cycle.
• RoDH-4 and DHRS9 are the only enzymes identified in humans that play a role in the formation of DHT and androstanedione.
• RODH4 has been described in the publication of T. Karlsson et al. (Biochem Biophys Res., 2003 Mar 28; 303 (1): 273-278), as an enzyme involved in the stimulation of sebum secretion in vitro by the transformation of 3-alphadihydrotestosterone and androsterone to dihydrotestosterone (DHT) and androstanedione.
• DHT dihydrotestosterone
• 13-cis retinoic acid (isotretinoin) well known for its anti-acne activity, is believed to reduce the formation of DHT and androstandione in vitro by inhibition of the RODH4 enzyme.
• RoDH4 is not expressed (mRNA) in the human sebaceous glands and weakly in the epidermis. This low presence of mRNA is confirmed using conventional RT-PCR on human epidermal biopsies. The other two subtypes RODH and RODH5 are undetectable in these two compartments of the skin.
• RDH11 is highly expressed in many different tissues with a greater proportion in the spinal cord, prostate, fetal brain, liver, kidney and testis. Unlike RDH11, DHRS9 is found in a much smaller number of organs (testis, heart, marrow and colon) with strong expression in the trachea.
• DHRS9 Hydrogenase Reductase Member 9
• DHRS9 Hydrogenase Reductase Member 9
• This observation is interesting insofar as it allows precisely to consider the activation or selective inactivation of this enzyme in the sebaceous glands unlike other subtypes of RoDH (including RODH4) expressed in humans. It therefore proposes to target the DHRS9 protein to prevent and / or treat acne, seborrheic dermatitis or any other skin disorder associated with hyperseborrhoea.
• Acne means all forms of acne, namely acne vulgaris, comedones, polymorphs, nodulocystic acnes, conglobata, or secondary acne such as solar acne, drug or professional.
• skin disorder associated with a hyperseborrhoea is meant in particular the appearance of oily and / or shiny skin, the presence of comedones, seborrheic dermatitis, the formation of dandruff.
• DHRS9 is expressed in different compartments of human skin tissue, but significantly more in the sebaceous glands than in the epidermis. Except otherwise, “DHRS9” refers to human DHRS9, the sequence of which is referenced in Genbank (NCBI) under accession number NM_005771.
• DHRS9 has no activity with respect to retinoids, but is regulated by them ("Characterization of a novel airway epithelial cell-specific short chain alcohol dehydrogenase / reductase gene whose expression is up- The pharmacokinetics of retinoids are described in Soref CM et al., J Biol Chem, 2001 Jun 29, 276 (26): 24194-202).
• DHRS9 inhibitor means any substance, simple or complex compound, of natural or synthetic origin, capable of inhibiting or reducing the activity or the expression of the DHRS9 enzyme. expression of its gene or the activity of at least one of its promoters, and / or capable of inhibiting, reducing or slowing down the reaction catalyzed by this enzyme. The inhibitor substantially eliminates or reduces the enzymatic activity of DHRS9.
• substantially means a reduction of at least 25%, preferably at least 35%, more preferably at least 50%, and more preferably at least 70% or 90%. More particularly, it may be a compound that interacts with, and blocks, the catalytic site of the enzyme, as compounds of the competitive inhibitory type.
• DHRS9 inhibitors examples include Citral or Carbenoxolone.
• Citral or lemonal is the name given to two isomers of the empirical formula C10H16O.
• the two components are diastereoisomers: the trans isomer is known as geranial or citral A.
• the cis isomer is known as the mineral or citral B.
• Géranial Néral Citral is the major constituent of lemongrass oil and other plants of the genus Cymbopogon. It is also present in verbena, orange or lemon oils.
• Carbenoxolone is a synthetic derivative of glycyrrizinic acid of formula below:
• the present invention thus relates to a method for the preventive and / or curative treatment of acne, seborrheic dermatitis or any skin disorder associated with hyperseborrhoea, which method comprises the administration of a therapeutically effective amount of a modulator. , preferably an inhibitor of the human enzyme DHRS9, to a patient in need of such treatment.
• a modulator preferably an inhibitor of the human enzyme DHRS9
• the subject of the invention is also the use of a modulator of DHRS9, for the preparation of a medicament for the preventive and / or curative treatment of acne, of seborrheic dermatitis or of skin disorders associated with hyperseborrhoea .
• the medicament according to the invention is intended for the preventive and / or curative treatment of acne, of seborrheic dermatitis.
• the invention finally relates to the cosmetic use of a modulator of the human enzyme DHRS9, for the aesthetic treatment of oily skin.
• Such a modulator preferably an inhibitor, of the enzyme DHRS9, is useful for preparing a medicament for the prevention and / or treatment of acne, seborrheic dermatitis or skin disorders associated with hyperseborrhoea.
• the medicament is effective in the treatment of acne or seborrheic dermatitis.
• This medication can be given orally, parenterally, or topically.
• such drug is intended for topical application. Topically, we mean an application on the skin or mucous membranes.
• the pharmaceutical composition may be in the form of tablets, capsules, dragees, syrups, suspensions, solutions, powders, granules, emulsions, suspensions of microspheres or nanospheres or lipid vesicles or polymers for controlled release.
• the pharmaceutical composition may be in the form of solutions or suspensions for infusion or for injection.
• the pharmaceutical composition is more particularly intended for the treatment of skin and mucous membranes and may be in the form of ointments, creams, milks, ointments, powders, soaked swabs, solutions, gels , sprays, lotions or suspensions. It may also be in the form of suspensions of microspheres or nanospheres or lipid or polymeric vesicles or polymeric patches or hydrogels allowing controlled release.
• This composition for topical application may be in anhydrous form, in aqueous form or in the form of an emulsion.
• the composition may comprise a modulator content of the DHRS9 enzyme ranging from 0.001 to 10% by weight, especially from 0.01 to 5% by weight relative to the total weight of the composition.
• the pharmaceutical composition may further contain inert additives or combinations thereof, such as wetting agents;
• UV-A and UV-B filters and antioxidants, such as alpha-tocopherol, butylhydroxyanisole or butylhydroxytoluene, superoxide dismutase, ubiquinol or certain metal chelators.
• antioxidants such as alpha-tocopherol, butylhydroxyanisole or butylhydroxytoluene, superoxide dismutase, ubiquinol or certain metal chelators.
• the present invention also relates to the use of the human DHRS9 enzyme as a research tool in acne, seborrheic dermatitis and / or in any skin disorder associated with hyperseborrhoea.
• This enzyme is a new tool for the identification, selection or characterization of a compound for the treatment of acne, seborrheic dermatitis and / or any skin disorder associated with hyperseborrhoea.
• the invention also relates to a method for diagnosing acne or predisposition to acne, comprising a step of measuring the activity of the DHRS9 enzyme in a biological sample, in particular by measuring the amount of DHT in said biological sample.
• the invention also relates to a diagnostic kit for acne comprising the enzyme DHRS9.
• a biological sample corresponds to any sample or sample of living organism, preferably a mammal, in particular a human organism, in an amount sufficient to be characterized.
• a biological sample may be a sample of skin, scalp, mucosa, or cell.
• the present invention relates to the use of the human DHRS9 enzyme for the identification, selection or characterization of a compound for the treatment and / or prevention of acne, seborrheic dermatitis. and / or any skin disorder associated with hyperseborrhoea.
• the subject of the invention is the use of the human DHRS9 enzyme for the identification of a compound intended to prevent and / or improve the symptoms of acne, seborrheic dermatitis and / or any disorder. skin associated with hyperseborrhea.
• the DHRS9 enzyme is used in the prevention and / or treatment of acne.
• the invention also relates to an in vitro method for screening candidate compounds for the preventive and / or curative treatment of a pathology chosen from acne, seborrheic dermatitis and cutaneous disorders associated with a hyperseborrhoea, comprising the determining the ability of a compound to inhibit the expression or activity of the human DHRS9 enzyme, or the expression of its gene or the activity of at least one of its promoters.
• the in vitro screening method described above comprises the following steps: a) preparing at least two biological samples; b) bringing one of the samples into contact with one or more test compounds; c) measuring the expression or the activity of the DHRS9 enzyme, the expression of its gene or the activity of at least one of its promoters, in the biological samples; d) selecting said compounds for which an inhibition of the expression or the activity of the DHRS9 enzyme, or an inhibition of the expression of its gene or an inhibition of the activity of at least one of its promoters is measured in the treated sample of step b) relative to the untreated sample.
• the biological samples are cells transfected with a reporter gene operably linked to all or part of the promoter of the gene coding for the enzyme DHRS9, and the step c) described above consists in measuring the expression of said reporter gene.
• the biological samples are cells expressing the gene coding for the DHRS9 enzyme, and the step c) described above consists in measuring the expression of said gene.
• the cell used here can be of any type. It may be a cell expressing the DHRS9 enzyme gene endogenously.
• expression of the DHRS9 enzyme gene or reporter gene can be determined by evaluating the transcription rate of said gene, or its translation rate.
• transcription rate of a gene is meant the amount of the corresponding mRNA produced.
• translation rate of a gene is meant the amount of protein produced.
• the invention also relates to an in vitro method for diagnosing or monitoring a revolution of acne, seborrheic dermatitis or skin disorder associated with hyperseborrhea in a subject, comprising comparing the expression or the activity of the enzyme DHRS9, the expression of its gene or the activity of at least one of its promoters, in a biological sample of a subject relative to a biological sample of a control subject.
• the invention also relates to an in vitro method for determining a subject's susceptibility to developing acne, seborrhoeic dermatitis or skin disorder associated with hyperseborrhoea, comprising comparing the expression or the activity of the enzyme
• DHRS9 the expression of its gene or the activity of at least one of its promoters, in a biological sample of a subject relative to a biological sample of a control subject.
• DHRS9 activity of the enzyme DHRS9
• promoter activity is meant the ability of this promoter to trigger the transcription of the coded DNA sequence downstream of this promoter (and thus indirectly the synthesis of the enzyme DHRS9).
• the activity of the enzyme DHRS9 can be evaluated by measuring the amount of DHT produced, or by measuring the amount of substrate, the 3alpha diol, which must disappear, in the samples.
• the present invention relates to a research tool comprising the human DHRS9 enzyme.
• the research tool may also include the analysis of the effects of certain treatments on the activity of the DHRS9.
• this research tool may be such that the activity of the enzyme DHRS9 is measured by the level of DHT produced.
• the invention finally relates to the cosmetic use of a modulator, preferably an inhibitor of the human enzyme DHRS9, for the aesthetic treatment of oily skin.
• the human DHRS9 enzyme is expressed in various tissues other than the skin, such as the colon, bone marrow or trachea, as shown in Example 1 and Figure 1.
• the DHRS- 9 is very poorly expressed in other tissues other than the skin, which is an important advantage, which allows avoid known systemic side effects related to changes in androgen metabolism. This weak expression is to oppose to another possible target: RoDH11 which is strongly expressed in many organs and which is ubiquitous.
• FIG. 3 represents a global diagram of the pathogenesis of the disease. acne, which includes the action of DHRS9.
• FIG. 4 represents the activity of conversion of 3 ⁇ -diol to DHT by DHRS9 in cells transfected or not by DHRS9.
• DHRS9 is a potential therapeutic target for the development of new therapies aimed at treating acne, seborrheic dermatitis or any skin disorder associated with hyperseborrhoea.
• the measurement was performed by real-time PCR.
• the expression of the target genes: hRoDH, RoDH11, RoDH-4, RoDH-5 and DHRS9 was conducted on different human tissues using 5 specific primers,
• DHRS9 is predominantly expressed in the colon, spinal cord and trachea.
• This experiment was performed from samples from two different patients. Dissections under binocular microscope and or by laser microdissection on Frozen tissue section was performed to isolate the epidermis and sebaceous glands. The amount of mRNA encoding different enzymes, including DHRS9, was then measured in each of these compartments.
• DHRS9 is expressed at least 20-fold more in the sebaceous glands than in the epidermis alone.
• the cDNA encoding human DHRS9 was subcloned into a pcDNA3.1 / zeo expression vector. This is then transfected into a cell line named PALM for PC-3 Androgen receptor Luciferase MMTV.
• This cell line is derived from PC-3 cells that have been stably transfected with the expression vector encoding the human androgen receptor (pSG5-puro-h androgen receptor (AR)) and the reporter gene vector. coding luciferase (pMMTV-neo-Luc) whose promoter has AR response elements.
• the conversion activity of 3 ⁇ -diol into DHT by the DHRS9 can thus be followed via the AR transactivation.
• the response dose of the 3 ⁇ -diol substrate gives an AC 50 for AR equal to 189nM for untransfected cells compared to 19 nM for cells transfected with DHRS9 (see Figure 4, 0 Transfected cells DHRS9, + Non-transfected cells).
• DHRS9 is able to shift I ⁇ C50 from the 3 ⁇ -Diol to the left by about a log of magnitude thus allowing the evaluation of inhibitors (see Figure 4, 0 Transfected cells DHRS9 , + Untransfected cells).

Landscapes

• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• General Health & Medical Sciences (AREA)
• Animal Behavior & Ethology (AREA)
• Engineering & Computer Science (AREA)
• Veterinary Medicine (AREA)
• Public Health (AREA)
• Dermatology (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Zoology (AREA)
• Wood Science & Technology (AREA)
• General Chemical & Material Sciences (AREA)
• Pharmacology & Pharmacy (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Medicinal Chemistry (AREA)
• Biophysics (AREA)
• Genetics & Genomics (AREA)
• Immunology (AREA)
• Microbiology (AREA)
• Molecular Biology (AREA)
• Analytical Chemistry (AREA)
• Physics & Mathematics (AREA)
• Biochemistry (AREA)
• General Engineering & Computer Science (AREA)
• Biotechnology (AREA)
• Birds (AREA)
• Epidemiology (AREA)
• Cosmetics (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
• Investigating Or Analysing Biological Materials (AREA)
• Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
• Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Applications Claiming Priority (2)

Publications (1)

Family

ID=39027085

Family Applications (1)

Country Status (5)

Family Cites Families (2)

• 2007
  • 2007-07-06 FR FR0756309A patent/FR2918388B1/fr not_active Expired - Fee Related
• 2008
  • 2008-07-07 CA CA2693153A patent/CA2693153A1/fr not_active Abandoned
  • 2008-07-07 WO PCT/FR2008/051269 patent/WO2009010687A2/fr active Application Filing
  • 2008-07-07 EP EP08826450A patent/EP2167677A2/de not_active Withdrawn
• 2010
  • 2010-01-05 US US12/652,283 patent/US20100168231A1/en not_active Abandoned

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events