WO2019023776A1 - Procédé d'évaluation d'hydratation bioactive d'un ingrédient ou d'un mélange d'ingrédients, compositions cosmétiques pour la modulation de gènes responsables de l'hydratation bioactive de la peau et méthode pour la modulation de l'expression de gènes responsables de l'hydratation bioactive de la peau - Google Patents

Procédé d'évaluation d'hydratation bioactive d'un ingrédient ou d'un mélange d'ingrédients, compositions cosmétiques pour la modulation de gènes responsables de l'hydratation bioactive de la peau et méthode pour la modulation de l'expression de gènes responsables de l'hydratation bioactive de la peau Download PDF

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Publication number
WO2019023776A1
WO2019023776A1 PCT/BR2018/050261 BR2018050261W WO2019023776A1 WO 2019023776 A1 WO2019023776 A1 WO 2019023776A1 BR 2018050261 W BR2018050261 W BR 2018050261W WO 2019023776 A1 WO2019023776 A1 WO 2019023776A1
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WO
WIPO (PCT)
Prior art keywords
skin
hydration
bioactive
ingredients
molecules
Prior art date
Application number
PCT/BR2018/050261
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English (en)
Portuguese (pt)
Inventor
Leonardo RODRIGUES DE PAULA
Original Assignee
Natura Cosméticos S.A.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Application filed by Natura Cosméticos S.A. filed Critical Natura Cosméticos S.A.
Publication of WO2019023776A1 publication Critical patent/WO2019023776A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • 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
    • 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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6804—Nucleic acid analysis using immunogens
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof

Definitions

  • the present invention relates to a method for characterizing bioactive (or active or dynamic or biological) hydration by employing techniques for assessing gene and protein expression which allows the identification of novel ingredients or a combination of efficient ingredients in the stimulus of the five endogenous mechanisms of the skin associated with said hydration.
  • the epidermis the outermost layer of the skin, is always renewing and its last layer, which makes contact with the outer medium is called stratum corneum.
  • EC comprises the final stage of epidermal cell differentiation and is specifically represented by dead and anucleated keratinized cells embedded in a lipid matrix acting as a barrier separating the external environment from internal homostasy.
  • the natural barrier formed by EC depends critically on its composition, represented by proteins (75-80%), lipids (5-15%) and other constituents (5-10%).
  • the lipid composition of human EC can vary quantitatively between individuals and body parts, but mainly comprises ceramides, fatty acids, cholesterol, esters, triglycerides and phospholipids. When these components are not properly balanced, the skin's ability to maintain water is reduced and the skin becomes more susceptible to of environmental factors, leading to an impaired barrier.
  • a major role in maintaining the integrity of the EC barrier is represented by water.
  • One of the main aspects is related to its ability to mediate the activity of many hydrolytic enzymes in the skin, including those responsible for proper desquamation of corneocytes, as well as those responsible for the formation of the NMF (Natural Moisturizing Factor) .
  • NMF Natural Moisturizing Factor
  • the epidermis is also responsible for the regeneration of the skin. Every 28 days a cell walks from the dermo-epidermal junction to the stratum corneum, then detached as a dead cell in the desquamation process. When the skin is dehydrated, this reconstitution and exchange are altered, making natural skin regeneration difficult.
  • hyaluronic acid present in the dermis and epidermal intercellular spaces. This molecule can bind up to a thousand times its weight with water molecules. In the dermis, along with other extracellular matrix molecules, it is responsible for holding water and maintaining adequate tissue drainage.
  • membrane protein channels called aquaporins are responsible for facilitating the transport of water and solutes, including glycerol and urea, between biological membranes. This conduction of water occurs through an osmotic gradient.
  • moisturizer it is recommended that adults use moisturizer on the whole skin at least once a day.
  • Such products generally include ingredients which typically act by two well known mechanisms: occlusion and wetting.
  • Occlusion hydration is promoted by ingredients that form a lipophilic film, which hinders the loss of water, increasing water retention in the stratum corneum.
  • Moisturizing by wetting is promoted by ingredients which retain water from the cosmetic composition, the atmosphere and the skin on the surface of the skin by means of a hydrophilic film formed on the horny layer, increasing the retention of water on the skin surface.
  • Dryness is one of the most present and chronic conditions of the skin, resulting in "stiffness" of the skin and damage to the skin in the form of cracks and cracks.
  • assessment of mechanical properties of the skin may be useful to understand how damage to dry skin occurs (eg, cracking).
  • the present invention relates to a method for characterizing bioactive hydration by employing techniques for assessing gene and protein expression which allows the identification of novel ingredients or combination of efficient ingredients in stimulating the five endogenous skin mechanisms associated with the said hydration.
  • bioactive hydration also referred to herein as biological hydration or biodynamic hydration or active hydration, is meant the ability of a balanced combination of ingredients to stimulate endogenous skin mechanisms to promote equilibrium between water and lipid content for efficient hydration and nutrition.
  • the endogenous mechanisms that determine bioactive hydration according to the present invention are simultaneously:
  • stimulating molecules that hold water in the deeper layers of the skin (dermis) such as hyaluronic acid and other components of the extracellular matrix
  • NMF Natural Moisturizing Factor
  • stimulating molecules carrying water and minerals to maintain the osmotic balance of the skin such as, for example, aquaporins.
  • the method for characterizing bioactive hydration is to evaluate the expression of an ingredient or combination of ingredients (complexes) in a set of genes associated with the five endogenous mechanisms of the skin that promote the balance between the hydric and lipid content for efficient hydration and nutrition, which characterize bioactive hydration.
  • CD44 CD44 molecule Indian blood group
  • SPTLC2 Serine palmitoyltransferase, long chain base subunit 2
  • TJP1 Tight junction protein 1
  • the set of genes associated with each endogenous mechanism according to the present invention is:
  • Stimulating molecules that retain water in the deeper layers of the skin (hyaluronic acid and other extracellular matrix components): BGN, CD44, COL1A1, COL1A2, COL4A4, DCN, ELN, FMOD, GPC3, HAS1, HAS2, HAS3, HMMR, HYAL1, HYAL2, LOX, LUM, MATN2, MMP12, MMP1, MMP9, SDC1, TIMP1, TIMP2, VCAN;
  • Stimulating molecules carrying water and minerals to maintain the osmotic balance of the skin AQP10, AQP1, AQP3, AQP5, CLDN1, CLDN4, CLDN7, GJA1, GJB2, TJP1.
  • the present invention also comprehensively contemplates cosmetic compositions for the modulation of genes responsible for bioactive hydration of the skin, either the body or the face, which comprises at least one ingredient or combination of ingredients that act simultaneously in the five endogenous mechanisms of the invention and less a cosmetically acceptable vehicle.
  • cosmetically acceptable excipients are those known to those skilled in the art for the composition of cosmetic bases in various forms, for example, emulsions, creams, gels, sils, and others known to those skilled in the art.
  • cosmetically acceptable excipients may be selected from those quoted in the International Cosmetic Ingredient Dictionary & Handbook, 16th Edition.
  • Example 1 Analysis of the efficacy in bioactive hydration of a potential ingredient
  • Gene and protein expression analysis was used to verify the modulation of the genes involved in the stimulation of the endogenous mechanisms of the skin, which promote the balance between water and lipid content for efficient hydration and nutrition, that is, bioactive hydration.
  • the sample was tested on human skin explants obtained after plastic surgery (blepharoplasty) and evaluated efficacy in modulating a total of 92 genes related to biological mechanisms determined to be relevant to skin hydration, as well as for some additional benefits such as integrity of the skin barrier, antioxidant, adhesion and cohesion between cells, cell renewal and anti-aging (elasticity, firmness, filling and dermo-epidermal cohesion).
  • Portion (1) was collected and subjected to total RNA extraction. From the RNA, the cDNA was prepared and this (50ng) was submitted to a real-time RT-PCR (Polymerase-Chain Reaction) step to evaluate the expression of 96 genes.
  • the gene expression profile and selection of the differentially expressed genes were performed with the aid of Expression Suite Software v.1.0.3 (Life Technologies).
  • the staining reaction was stopped by adding 2N H2 SO4 and the reading was performed on a microplate reader at 450 nm. Protein levels were expressed in pg / ml or g / ml and calculated from reference values obtained with a standard curve constructed at known concentrations of the protein. For statistical analysis, analysis of variance (ANOVA) was used. In all groups studied, those whose P values were less than 0.05 were considered statistically significant.
  • ANOVA analysis of variance
  • genes that had their expression evaluated by the study were: ACACA, ACACB, AQP1, AQP3, AQP5, ASAM, BGN, CASP14, CD44, CDH1, CDSN, CLDN1, CLDN4, CLDN7, COL1A1, COL1A2, COL4A4, COL7A1, CRNN, CST6, CTSB, CTSD, CTSE, CTSL, CTSV, DCN, DSC1, DSC2, DSG1, DSG3, DSG4, DSP, ELN, ELOVL3, EVPL, FASN, FLG, FMOD, GBA, GJA1, GJB2, HAL, HAS1, HAS2, HAS3, HMGCR, HMGCS1, HYAL1, HYAL2, ITGA6, ITGB4, IVL, KLF4, KLK5, KLK7, KRT1, KRT10, LAMA5, LCE2B, LOR, LOX, LUM, MATN2, MMP1, MMP12, MMP9, OCLN, PADI1, PADI1, PADI3,
  • Figure 1 shows the gene expression results obtained from a potential active ingredient compared to other market ingredients.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Analytical Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Organic Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • Pathology (AREA)
  • Biomedical Technology (AREA)
  • General Physics & Mathematics (AREA)
  • Urology & Nephrology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Dermatology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Birds (AREA)
  • Epidemiology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Hematology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Genetics & Genomics (AREA)
  • General Engineering & Computer Science (AREA)
  • Cell Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biophysics (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Cosmetics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne une méthode de caractérisation d'hydratation bioactive (ou active, dynamique ou biologique) par utilisation de techniques d'évaluation d'expression génique et protéique permettant d'identifier de nouveaux ingrédients ou une combinaison d'ingrédients efficaces dans la stimulation des cinq mécanismes endogènes de la peau associés à ladite hydratation.
PCT/BR2018/050261 2017-07-31 2018-07-27 Procédé d'évaluation d'hydratation bioactive d'un ingrédient ou d'un mélange d'ingrédients, compositions cosmétiques pour la modulation de gènes responsables de l'hydratation bioactive de la peau et méthode pour la modulation de l'expression de gènes responsables de l'hydratation bioactive de la peau WO2019023776A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
BRBR102017016427-6 2017-07-31
BR102017016427-6A BR102017016427A2 (pt) 2017-07-31 2017-07-31 Método para avaliar hidratação bioativa de um ingrediente ou mistura de ingredientes, composições cosméticas para a modulação de genes responsáveis pela hidratação bioativa da pele e método para a modulação da expressão de genes responsáveis pela hidratação bioativa da pele

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WO2019023776A1 true WO2019023776A1 (fr) 2019-02-07

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AR (1) AR112771A1 (fr)
BR (1) BR102017016427A2 (fr)
WO (1) WO2019023776A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113203718A (zh) * 2021-05-13 2021-08-03 桂林电子科技大学 一种基于荧光共振能量转移的gpc3检测方法

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015031971A2 (fr) * 2013-09-09 2015-03-12 Natura Cosméticos S.A. Composition comprenant un extrait de guaçatonga et un extrait d'aroeira, son utilisation, et méthode de prévention et/ou de traitement des signes provoqués par le viellissement cutané
WO2017201597A1 (fr) * 2016-05-24 2017-11-30 Natura Cosméticos S.A. Composition pour la modulation de gènes responsables des fonctions générales de la peau, méthode pour la modulation de l'expression de gènes responsables des fonctions générales de la peau, et utilisation d'un extrait végétal

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015031971A2 (fr) * 2013-09-09 2015-03-12 Natura Cosméticos S.A. Composition comprenant un extrait de guaçatonga et un extrait d'aroeira, son utilisation, et méthode de prévention et/ou de traitement des signes provoqués par le viellissement cutané
WO2017201597A1 (fr) * 2016-05-24 2017-11-30 Natura Cosméticos S.A. Composition pour la modulation de gènes responsables des fonctions générales de la peau, méthode pour la modulation de l'expression de gènes responsables des fonctions générales de la peau, et utilisation d'un extrait végétal

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113203718A (zh) * 2021-05-13 2021-08-03 桂林电子科技大学 一种基于荧光共振能量转移的gpc3检测方法
CN113203718B (zh) * 2021-05-13 2022-05-31 桂林电子科技大学 一种基于荧光共振能量转移的gpc3检测方法

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Publication number Publication date
AR112771A1 (es) 2019-12-11
BR102017016427A2 (pt) 2019-03-26

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