EP3902518A1 - Extrait de chlamydomonas acidophila, son procede de preparation et les compositions cosmetiques, et dermatologiques le comprenant - Google Patents
Extrait de chlamydomonas acidophila, son procede de preparation et les compositions cosmetiques, et dermatologiques le comprenantInfo
- Publication number
- EP3902518A1 EP3902518A1 EP19835309.6A EP19835309A EP3902518A1 EP 3902518 A1 EP3902518 A1 EP 3902518A1 EP 19835309 A EP19835309 A EP 19835309A EP 3902518 A1 EP3902518 A1 EP 3902518A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- extract
- advantageously
- skin
- acidophila
- composition
- Prior art date
- Legal status (The legal status 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 status listed.)
- Pending
Links
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- 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
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A61K8/9722—Chlorophycota or Chlorophyta [green algae], e.g. Chlorella
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/02—Algae
- A61K36/05—Chlorophycota or chlorophyta (green algae), e.g. Chlorella
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- A—HUMAN NECESSITIES
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- A61K38/00—Medicinal preparations containing peptides
- A61K38/01—Hydrolysed proteins; Derivatives thereof
- A61K38/011—Hydrolysed proteins; Derivatives thereof from plants
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- A—HUMAN NECESSITIES
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- 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
- A61K8/645—Proteins of vegetable origin; Derivatives or degradation products thereof
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- A—HUMAN NECESSITIES
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- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
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- 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
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
Definitions
- the invention relates to a peptide extract of the microalga Chlamydomonas acidophila and to a cosmetic, dermatological or pharmaceutical composition comprising such an extract.
- the invention also relates to a method of extracting a peptide extract of Chlamydomonas acidophila, as well as the extract capable of being obtained by said method.
- the invention also relates to a composition or such an extract for its use in the prevention or treatment of disorders or pathologies of the skin, mucous membranes or integuments, for its use in the prevention or treatment of vascular disorders, or also for its use in the prevention or treatment of adipose tissue alterations.
- the invention relates to a cosmetic care process for the skin, integuments or mucous membranes, with a view to improving their condition or their appearance, consisting in administering such a composition or such an extract.
- Micro-algae are single-celled, eukaryotic organisms, endowed with photosynthesis and which are therefore capable, like higher plants, of using CO2 from the air for their metabolism but also other nutrients such as phosphorus, nitrates, etc. They were among the first species to colonize the earth. There are around 30,000 species described but it is assumed that there are many more. Microalgae are found in their natural state, both in fresh and brackish to salt water around the world.
- Microalgae can be cultivated according to methods known to those skilled in the art, such as photo-reactors in environments controlled in light, pH and nutrients, and they have many outlets. Like the higher organisms, they are capable of synthesizing proteins, carbohydrates and lipids. Certain lipids are specific such as complex fatty acids or pigments with specific biological properties (xanthophylls). They have experienced a particular craze for a possible production of biofuel and their production in bioreactor has expanded. The other outlets are diverse: food for fish (aquarium and fish farms), food and human health (asthaxanthin extracted from Haematococcus pluvialis, spirulina proteins) and some outlets in the cosmetic industry.
- Chlamydomonas acidophila of the class Chlorophyceae (Family: Chlamydomonadaceae) is a green freshwater microalgae that proliferates in very acidic waters (pH 2.3 to 3.4) and adapted to environments loaded with heavy metals. In particular, it was identified and removed for the first time from volcanic lakes in Argentina. It is said to be rich in phytochelatins, special structures capable of chelating metals and in carotenoids (beta-carotene, lutein). Apart from publications, concerning its possible conditions of culture and development (tolerance to extreme pH, and heavy metals), there is little about its composition and its use. Its “cousin” Chlamydomonas reinhardii is used as a model organism in various scientific fields such as genetics.
- the subject of the invention is a peptide extract of the microalga Chlamydomonas acidophila.
- peptide extract is meant, within the meaning of the present invention, an extract comprising predominantly peptides.
- peptide is meant, within the meaning of the present invention, a polymer of amino acids linked together by peptide bonds.
- a peptide is in particular characterized by a molecular mass of between 200 and 10,000 Daltons (Da), limits included.
- the extract of Chlamydomonas acidophila according to the invention comprises at least 20% by weight of peptides, the percentages being expressed relative to the total weight of said extract.
- the extract according to the invention comprises from 20% to 90%, advantageously from 20% to 75%, more advantageously from 30% to 70%, typically 65%, by weight of peptides, the percentages being expressed relative to the total weight of said extract.
- the extract of Chlamydomonas acidophila according to the invention is substantially free of any protein, in particular any residual native protein. This makes it possible in particular to avoid allergic reactions, and to improve the solubility and the bioavailability of the extract according to the invention.
- protein is meant, within the meaning of the present invention, biological macromolecules formed from one or more polypeptide chains. Each of these chains consists of the chain of amino acid residues linked together by peptide bonds.
- a protein is notably characterized by a molecular mass greater than 10,000 Daltons (Da).
- the extract of Chlamydomonas acidophila according to the invention is substantially free of free amino acids. Free amino acids have a molecular weight of less than 200 Da.
- the peptides advantageously have a molecular mass of less than 3500 Daltons (Da).
- these peptides cover all of the amino acid-based compounds initially present in the extract.
- At least 80%, more advantageously at least 90%, of the peptides have a molecular mass of less than 1000 Da.
- At least 30% of the peptides more advantageously at least 35%, more advantageously at least 40% of the peptides, have a molecular mass of less than 500 Da.
- the distribution of the molecular weights of the peptides is expressed as a percentage relative to the total concentration of peptides.
- the peptide extract of Chlamydomonas acidophila is advantageously obtained by enzymatic hydrolysis, more advantageously in the presence of at least one protease.
- the extract according to the invention is more advantageously obtainable by the process described later in the description.
- the invention also relates to a process for the preparation of a peptide extract of the microalga Chlamydomonas acidophila, comprising at least one step of enzymatic hydrolysis.
- This step is advantageously carried out under the optimal conditions of pH and temperature, known to those skilled in the art, in particular under the optimal conditions of pH and temperature linked to the enzyme used.
- said enzymatic hydrolysis step is carried out in the presence of at least one protease.
- Said protease can advantageously be an alkaline protease or an acid protease, advantageously it is an alkaline protease.
- the process for preparing a peptide extract of Chlamydomonas acidophila comprises at least the following steps:
- step a) recovery of the peptide extract at the end of step c).
- the aqueous phase is advantageously water.
- the content of Chlamydomonas acidophila microalga in the aqueous dispersion is advantageously between 0.1% and 20%, more advantageously 1% and 10%, equivalent dry extract of the microalga.
- the enzymatic treatment (step b) is advantageously carried out by adding at least one protease, advantageously under the optimal pH and temperature conditions known to those skilled in the art, for example at a pH between 3.0 and 9.0 and typically at a temperature between 20 ° C and 90 ° C.
- the enzymatic treatment comprises the addition of an alkaline or acidic protease, advantageously an alkaline protease.
- the enzymatic hydrolysis step of the process according to the invention is very important, since it allows the native proteins present in Chlamydomonas acidophila to be transformed or "cut out" to obtain peptides.
- the enzymatic hydrolysis step is advantageously followed by a heat treatment step in order to denature the enzymes.
- This heat treatment step is advantageously carried out at a temperature above 40 ° C, typically between 80 ° C and 100 ° C.
- step d) the peptide extract is advantageously recovered by extraction of the dispersion obtained at the end of step c), advantageously with stirring, and advantageously at a pH of between 3.0 and 9.0 and at a temperature between 20 ° C and 90 ° C.
- the method comprises an additional filtration or centrifugation step, located between steps c) and d), optionally followed by an ultrafiltration, diafiltration, or nanofiltration.
- the filtration or centrifugation steps, optionally followed by ultrafiltration or diafiltration on membranes make it possible in particular to remove the residual proteins.
- the nanofiltration stages make it possible in particular to remove mineral salts or free amino acids for example.
- the method according to the invention advantageously comprises an ultrafiltration step at 15 kDa, advantageously between 10 and 15 kDa, carried out between steps c) and d), which makes it possible to eliminate any residual protein which is potentially allergenic.
- the method according to the invention also comprises a nanofiltration step with, for example, a cutoff threshold of between 100 Daltons and 300 Daltons, advantageously between 130 and 300 Daltons, typically between 200 Daltons and 300 Daltons, to remove part of the acids. amino or mineral salts, following the ultrafiltration stage.
- a nanofiltration step is carried out on a 200 Da membrane.
- the aqueous hydrolyzate obtained, ie the peptide extract according to the invention, can then be physically and microbiologically stabilized by adding a solvent such as glycerol or glycols like 1,3-propanediol in different proportions adapted to such stabilization.
- a solvent such as glycerol or glycols like 1,3-propanediol
- the glycerol will be present alone or in combination with water or a glycol, advantageously in a proportion of between 40 and 95% and preferably between 50 and 90%, by mass relative to the total mass of the peptide extract and solvent.
- the glycol and preferably 1,3-propanediol will advantageously be present alone or in combination with water or glycerol, advantageously in a proportion of between 40% and 95% and preferably between 50% and 90%, in mass relative to the total mass of the peptide extract and the solvent.
- the present invention further relates to a composition comprising the peptide extract of Chlamydomonas acidophila according to the invention, a solvent chosen from glycerol, glycols and their mixtures in an amount effective for a physical and microbiological stabilization action, and optionally some water.
- the effective amounts for a physical and microbiological stabilization action are as described above.
- the peptide extract can be stabilized by drying, by methods known to those skilled in the art, in the presence or absence of a support of the type, for example, maltodextrins or acacia fibers.
- a support of the type for example, maltodextrins or acacia fibers.
- the content of support typically varies according to a ratio ranging from 0% to 80% of support relative to the percentage of dry matter obtained in the liquid form of the extract.
- the extract can be dried by atomization, lyophilization or any process known to those skilled in the art and is preferably dried by lyophilization in order to obtain a final powder.
- the final powder advantageously comprises 30% to 70% by weight of dry matter of the extract, the complement to 100% being the lyophilization support. More advantageously, the final powder comprises 50% of dry matter obtained from the extract and 50% of lyophilization support, said lyophilization support preferably being of maltodextrin or acacia fiber type.
- the peptide extract can be obtained according to the following method:
- said microalgae used as a raw material comes from a culture in photobioreactor, in particular in photobioreactor in an agitated medium.
- said microalgae used as raw material comes from a culture in horizontal tubular photobioreactors ventilated in waves in an agitated medium, such as for example those developed by the company Microphyt and in particular described in patent application FR 2 943 685 and international application WO 2011/058267.
- the present invention also relates to an extract of Chlamydomonas acidophila, capable of being obtained by the process mentioned above.
- Such an extract meets the specifications defined above
- the subject of the invention is also a cosmetic, dermatological or pharmaceutical composition
- a cosmetic, dermatological or pharmaceutical composition comprising a peptide extract of Chlamydomonas acidophila, as active ingredient, and if appropriate an appropriate excipient.
- the peptide extract of Chlamydomonas acidophila is as defined above or is capable of being obtained by the process mentioned above.
- said extract is advantageously as defined in the above paragraphs relating to the extract according to the invention as such or those relating to the extract capable of being obtained by the process according to the invention.
- composition is advantageously formulated to be administered by external topical, vaginal or oral route.
- the composition according to the invention comprises from 0.001 to 10%, advantageously 0.01 to 5%, of said peptide extract of Chlamydomonas acidophila, by weight expressed as dry extract, relative to the total weight of the composition.
- composition according to the invention can also comprise one or more other active ingredients.
- the various preparations are suitable for topical administration and include in particular creams, emulsions, milks, ointments, lotions, oils, aqueous or hydro-alcoholic or glycolic solutions, powders, patches, sprays, shampoos, varnishes or any other product for external application.
- the various preparations notably include intimate hygiene care, oral care, such as, for example, toothpaste, oral solutions, gingival gels.
- the composition according to the invention can also comprise at least one cosmetically, pharmaceutically or dermatologically acceptable excipient.
- composition according to the present invention can also comprise at least one cosmetically, pharmaceutically or dermatologically known adjuvant known to those skilled in the art, chosen from surfactants, thickeners, preservatives, perfumes, dyes, chemical filters or minerals, moisturizers, thermal waters, etc.
- cosmetically, pharmaceutically or dermatologically known adjuvant known to those skilled in the art, chosen from surfactants, thickeners, preservatives, perfumes, dyes, chemical filters or minerals, moisturizers, thermal waters, etc.
- compositions according to the invention can be determined according to the criteria generally taken into account in the establishment of a pharmacological, dermatological or cosmetic treatment suitable for a patient or an animal, such as for example the age or body weight of the patient or animal, the severity of his general condition, the tolerance to treatment, the side effects observed, the type of skin.
- the subject of the invention is also an extract according to the invention or extract capable of being obtained by the process according to the invention or a composition according to the invention, for its use for preventing and / or treating:
- the subject of the invention is also the use of an extract according to the invention or of an extract capable of being obtained by the process according to the invention or of a composition according to the invention, for the manufacture of a cosmetic, pharmaceutical or dermatological composition for preventing and / or treating:
- the invention further relates to a method for preventing and / or treating: - disorders or pathologies of the skin and / or mucous membranes (for example the gums, periodonts, genital mucosa) and / or integuments (for example the hair and nails);
- - disorders or pathologies of the skin and / or mucous membranes for example the gums, periodonts, genital mucosa
- / or integuments for example the hair and nails
- composition according to the invention comprising the administration, in particular the topical administration, of an effective amount of an extract according to the invention or of an extract capable of being obtained by the process according to the invention or of a composition according to the invention , to a subject in need.
- the extract according to the invention or the extract capable of being obtained by the process according to the invention or the composition according to the invention is intended for the prevention and / or treatment of allergic, inflammatory reactions or pathologies , irritants or disorders of the barrier or homeostasis of the skin, appendages (hair and nails) and / or mucous membranes (gums, periodonts, genital mucosa) immature (normal) or mature (s) ) / aged (s).
- composition or the extract according to the invention can be used for the prevention and / or the treatment of reactions, disorders or pathologies:
- mucous membranes such as the gums and periodonts which can present gingivitis (sensitive gums of newborns, hygiene problems, due to smoking or others), periodontopathies, or genital mucous membranes which can present irritations of the external male or female genital spheres or internal and / or
- integuments such as nails (brittle, fragile nails ...) and hair (alopecia, dandruff, hirsutism, seborrheic dermatitis, folliculitis) immature, normal or mature, presenting in particular scalp disorders such as alopecia (or alopecia areata) androgenetic, acute, localized, scarring, congenital, occipital of the infant, aerata, due to chemotherapy / radiotherapy or telogen effluvium, anagen effluvium, hair dystrophy, trichotillomania, ringworm or oily or dry dandruff.
- the invention also relates to a method of cosmetic care of the skin and / or the integuments and / or mucous membranes, with a view to improving their condition and / or their appearance, consisting in administering an extract according to the invention or an extract capable to be obtained by the process according to the invention or a composition according to the invention.
- the cosmetic care process makes it possible to firm the skin and to reduce the “orange peel” effect advantageously topically on the skin and / or integuments and / or mucous membranes.
- the invention relates to a cosmetic care process for the skin and / or integuments, to act on the elasticity or firmness of the skin, in particular as a tightening or anti-wrinkle agent, to act on sensitive skin. , or also to act against pollution, consisting in applying to the skin and / or the integuments a composition or an extract according to the present invention.
- the invention relates to a cosmetic care process for the skin and / or the appendages, with a view to preventing alterations of the barrier and its dehydration, consisting in applying to the skin and / or the appendages a composition or an extract according to the present invention.
- the invention relates to a cosmetic skin care process, with a view to preventing aging thereof, consisting in applying to the skin a composition or an extract according to the present invention.
- composition or extract according to the present invention can also be advantageously used in the prevention and / or treatment of vascular disorders, in particular redness and rosacea.
- composition or extract according to the present invention can also be advantageously used in the prevention and / or treatment of alterations in adipose tissue.
- the adipose tissue alterations are in particular cellulite or the "orange peel” effect.
- the composition according to the invention makes it possible to firm the skin.
- Figure 1 represents the measurement results of erythema intensity: Active / Placebo / Untreated Zone comparisons. NS: non-significant difference. *: p ⁇ 0.05 (Example 3B).
- Figure 2 shows the evolution over time of the blood flow (Example 3B).
- FIG. 3 shows the illustrations of the PIE results obtained at D0 and D28 (Example 3C).
- Figure 4 shows the illustrations of the hydration results obtained at D0 and D28 (Example 3C).
- Figure 5 shows the illustrations of the quantity of NMFs quantified at D0 and D28 (Example 3C).
- Figure 6 represents the illustrations of the quantity of ceramides quantified with DO and D28 (Example 3C).
- Figure 7 shows the illustrations of the amount of ILIRA quantified at DO and D28 (Example 3C).
- Figure 8 shows the illustrations of the Nile red / lnvolucrine ratio at DO and D28 (Example 3C).
- Example 1 extract according to the invention
- a peptide extract is obtained according to the following process:
- Example 2 Tests of biological activities of the extract according to the invention in vitro) The biological activity of the extract of Chlamydomonas acidophila (CAP) obtained in Example 1 was demonstrated in vitro as described below.
- CAP Chlamydomonas acidophila
- CAP Chlamydomonas acidophila
- the GBA / GBAP1 gene encoding the enzyme Glucosylceramidase or b-glucocerebrosidase is over expressed after 6 hours of treatment with CAP.
- RAB11A codes for a GTPase (Ras-related protein Rab-llA) involved in the biogenesis of lamellar bodies in keratinocytes. This underlines the importance of RAB11A in the homeostasis of the epidermal barrier.
- GTPase Ras-related protein Rab-llA
- Hyaluronan synthases-2 and -3 are enzymes responsible for the synthesis of hyaluronic acid (HA).
- the PADI1, BLMH, CASP14 genes code for enzymes involved in the production of natural hydration factors (or NMF for Natural Moisturizing Factors).
- Table 2 below presents the most significant results of the CAP extract on the expression of genes in fibroblasts.
- Table 2 Variations in the expression of genes of interest in normal human dermal fibroblasts (NHDFs) Relative quantity (QR) compared to the equal control 1 / p value determined following a Student test
- HAS1, HAS2 5.76 0.008
- HAS3, produced by fibroblasts in the dermis are responsible for the biosynthesis of hyaluronic acid (AH).
- NAD (P) H dehydrogenase, quinone 1 (NQOl) is a cytosolic flavoprotein under the control of the transcription factor Nrf-2. NQOl promotes, by reduction, the formation of hydroquinones from quinones, preventing the production of radical species.
- the HSP70 protein encoded by the HSPA1A gene
- is a chaperone molecule heat shock protein 70 which inhibits the aggregation of denatured proteins, promotes their renaturation (refolding) and controls essential mediators of the apoptotic machinery.
- the thioredoxin system is one of the major antioxidant defense systems. Among the 3 isoforms of thioredoxine reductase, isoform 1 encoded by the TXNRD1 gene regulated by the CAP extract is the most studied.
- peroxyredoxin-6 PRDX6
- PRDX6 peroxyredoxin-6
- the CAP extract stimulates the GLOl gene, which is part of the GLO system which detects and neutralizes certain carbonyls, and thus prevents them from attacking cells and their components.
- the CAP extract stimulates the expression of PSMB1, a gene coding for the beta 1 subunit of the proteasome.
- the proteasome plays a major role in the maintenance of protein homeostasis by eliminating damaged or malformed proteins that could alter cell functioning.
- LOXL2 The gene expression of LOXL2 is also increased, this gene is part of the family of Lysyl oxidases, enzymes involved in the assembly of elastin and collagen fibers.
- the inflammatory response is the normal, immediate and transient response of the body to any attack on the environment.
- the keratinocyte is one of the first cells participating in the initiation of the inflammatory reaction in response to an attack on the environment.
- PGE prostaglandins
- the anti-inflammatory activity of the extract of Chlamydomonas acidophila according to the invention was evaluated on a model of inflammation induced on keratinocytes by treatment with PMA (Phorbol 12-Myristate 13-Acetate). The release of TNF ⁇ and Prostaglandin E2 (PGE2) cytokines was analyzed.
- PMA Phorbol 12-Myristate 13-Acetate
- PGE2 Prostaglandin E2
- the inflammation was then induced by adding PMA at 10 pg / mL overnight.
- the PMA at 10 pg / ml significantly increased the release of TNF ⁇ into the supernatants of keratinocytes and therefore induced inflammation well.
- 0.1 pM dexamethasone and 0.1 pM indomethacin, 24 h pretreatment significantly reduced the release of TNFa, thus demonstrating their anti-inflammatory effect and validating the test.
- the extract of Chlamydomonas acidophila in 24h pre-treatment at different concentrations, significantly reduced the release of TNFa and therefore showed an anti-inflammatory action against PMA.
- Nickel is the major cause of allergic contact dermatitis in the population, with a global prevalence of around 8.6%
- the objective of the study described below is to assess the effect of the CAP extract on the release of IL8 by nickel-stimulated keratinocytes.
- Normal human epidermal keratinocytes were pretreated for 24 hours with the extract of CAP at 0.01% and 0.05% of dry matter or with the anti-inflammatory reference molecule Dexamethasone at 1 mM.
- the keratinocytes were then treated for 24 hours with Nickel: NiS0 4 at 10 mM.
- the amount of IL8 produced by the cells was evaluated by an ELISA test in the supernatants.
- the IL8 concentration assayed was normalized with respect to the amount of total intracellular proteins evaluated by BC Assay.
- the CAP extract induces a significant reduction in the release of IL8 induced by nickel stress in keratinocytes.
- the extract of Chlamydomonas acidophila inhibits the release of a major cytokine, IL8, against the background of inflammatory stress induced by Nickel.
- the extract is therefore of interest in the context of contact allergy or skin hypersensitivity linked by Nickel.
- the objective of this study is to evaluate the anti-inflammatory activity of the extract Chlamydomonas acidophila (CAP) vis-à-vis a stress with heavy metals, represented by Cadmium, on normal human keratinocytes.
- CAP Chlamydomonas acidophila
- the PGE2 concentration assayed was normalized with respect to the amount of total intracellular proteins evaluated by BC Assay.
- the CAP extract induces a decrease in the release of PGE2 induced by Cadmium stress in keratinocytes.
- the extract of Chlamydomonas acidophila inhibits the production of Prostaglandin E2 (PGE2) induced by Cadmium stress.
- the extract therefore provides protection for the skin against heavy metal stress, for example in the context of environmental pollution.
- the anti-inflammatory activity of the extract of Chlamydomonas acidophila was evaluated on a model of inflammation induced by treatment with SDS (Sodium Dodecyl Sulfate) on reconstituted epidermis.
- Reconstructed Human Epidermis were preincubated for 24 hours in the presence of 0.01% CAP and 0.05% dry matter. 0.025% SDS was then applied to the surface of the epidermis which was again incubated for 24 hours, still in the presence of the CAP extract.
- TNF ⁇ cytokine Tumor Necrosis Factor alpha
- the gene expression of inflammatory markers and of the barrier was evaluated by qRT-PCR.
- the CAP extract significantly inhibited the overproduction of TNF ⁇ and increased the expression of keratin 1.
- the antioxidant effect of the active ingredient is evaluated by the incorporation of DCFH-DA (2 ′, 7 ′ - Dichlorofluorescin diacetate) in cultured keratinocytes.
- DCFH-DA (2 ′, 7 ′ - Dichlorofluorescin diacetate)
- This molecule is a non-fluorescent marker in the non-oxidized state.
- oxidative conditions here H2O2 stress
- DCFH-DA will be degraded into DCF, a molecule which will emit fluorescence.
- the fluorescence thus measured will be proportional to the quantity of reactive oxygen species produced by the cell in the presence of H2O2 and / or of the extract.
- the cells are then treated for 1 h in the presence of DCFH-DA at 0.5 mM. Oxidation is induced by adding H2O2 at 100 mM for 20 minutes. A second treatment with the tested products is carried out simultaneously with H2O2 stress (at the same concentrations as the pre-treatment).
- DFU fluorescence density
- the extract of Chlamydomonas acidophila has demonstrated an antioxidant effect against stress induced by H 2 0 2 .
- the gene expression of the markers of the barrier function and of the hydration was evaluated by qRT-PCR.
- the extract of Chlamydomonas acidophila also stimulated the expression of the markers SLC6A6 and SLC5A3, coding for TAUT (taurine membrane transport channel) and SMIT (myoinositol transport channel) respectively.
- the extract of Chlamydomonas acidophila modulates the inflammation induced by Th2 stress in keratinocytes by inhibiting the gene expression of the chemoattractant factors CCL5 and CCL27.
- the basophil activation test was carried out using the Flow CAST ® kit (BÜHLMANN, ref. FKCCR).
- the stimulant, fM LP at 1 mM was then added and the blood was incubated for an additional 15 minutes in the presence of the labeling buffer containing a mixture of monoclonal antibodies (anti-CD63-FITC and anti-CCR3-PE).
- Cromoglycate a known anti-allergic, the mechanism of action of which involves stabilizing the plasma membrane at which it inhibits intracellular penetration of Ca ++ , this ion being essential for mast cell degranulation.
- SB202190 tested at 30 mM, as well as cromoglycate, tested at 10 mM, both showed a significant inhibitory effect on the activation of basophils induced by fMLP (respectively, 26% and 46% inhibition).
- the CAP extract tested at 0.033% and 0.1%, showed a clear and concentration-dependent inhibitory effect on the activation of basophils induced by fMLP (respectively, 22% and 39% inhibition).
- Chlamydomonas acidophila extract inhibits the activation of basophils.
- Chlamydomonas extract acidophila could help modulate the processes involved in initiating the allergic response.
- the CAP active ingredient (3% active ingredient) has demonstrated significant effectiveness on the following parameters:
- the intensity of blood flow measured by TiVi is significantly lower over the area treated with the active ingredient compared to the untreated area.
- the redness measured by spectrocolorimetry is significantly lower on the area treated with the active ingredient compared to the untreated area.
- the decrease in the intensity of the blood flow measured by Tivi is significantly greater over the area treated with the active ingredient compared to the untreated area.
- the reduction in redness measured by spectrocolorimetry is significantly greater in the area treated with the active ingredient compared to the untreated area.
- the CAP active ingredient (3% active ingredient) has demonstrated significant effectiveness on the following parameters: Instrumental measurements
- the people recruited for this study are subjects:
- the method used by the TiVi 700 is based on the fact that green light is strongly absorbed by the cells of blood vessels, while red light is moderately absorbed.
- the method eliminates specular reflection to take into account only the light returned by the skin tissue.
- the device thus produces an intensity map with each pixel representing a concentration of blood cells in the skin.
- a decrease in the intensity of the concentration of blood cells in the skin indicates an anti-inflammation effect.
- the graphs of Figures IA, IB and IC represent the pair-to-pair comparisons between the active, the placebo and the untreated zone on the intensity of the erythema 20 minutes after application of Methyl-Nicotinate measured by TiVi.
- the ordinate parameter represents the intensity of the blood flow (concentration of red blood cells).
- Tables 13A and 13B below (Tables 13A and B: mean and standard deviation of the measurements with% of subjects with a positive effect% difference and p value (exact value of significance).
- the graph Figure 2 shows the evolution over time of the blood flow values.
- the results show that the active ingredient significantly reduces the intensity of the erythema created compared to the untreated area.
- the intensity of the erythema on the placebo area does not differ from the area treated.
- the intensity of the erythema on the active area is significantly lower compared to placebo.
- the people recruited for this study are subjects:
- the skin barrier helps regulate water loss through evaporation. When this barrier is damaged, the insensible water loss increases. Conversely, a reinforced barrier corresponds to an insensible loss of water less.
- the insensible water loss was measured by a Tewameter TM 300.
- the principle is to measure the temperature and the relative humidity in a tube with one of the openings applied to the skin by 2 sensors located at 2 different heights. Fick's law is then used to determine the insensible water loss.
- Table 14 values at D0 and D28 of the insensible water loss. Evolution percentages and statistics.
- the hydration of the skin was measured by a CM 825 corneometer. This device is based on the principle of capacitance measurement, allowing a measurement of the hydration of the surface layers of the skin (10 to 20 ⁇ m of depth).
- Table 15 values at D0 and D28 of hydration. Evolution percentages and statistics.
- NMFs Natural Hydration Factors
- the quantity of NMFs includes urocanic acid (UCA), Pyrrolidone carboxylic acid (PCA) and serine.
- UCA urocanic acid
- PCA Pyrrolidone carboxylic acid
- serine serine
- Ceramides by Liquid Chromatography coupled with detection by mass spectroscopy (LC / MS).
- the amount of ceramides includes ceramides with bases [S], [DS] and [P]
- the content of ceramides provides information on the state of the skin barrier.
- Table 16 values at DO and D28 of the quantity of NMFs. Evolution percentages and statistics.
- Table 17 values at DO and D28 of the quantity of ceramides. Evolution percentages and statistics.
- Table 18 values at DO and D28 of the amount of ILIRA. Evolution percentages and statistics.
- Table 19 DO and D28 values of the Nile red / lnvolucrine ratio. Evolution percentages and statistics.
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PCT/EP2019/087171 WO2020136283A1 (fr) | 2018-12-28 | 2019-12-30 | Extrait de chlamydomonas acidophila, son procede de preparation et les compositions cosmetiques, et dermatologiques le comprenant |
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WO2024181027A1 (fr) * | 2023-03-02 | 2024-09-06 | 本田技研工業株式会社 | Composition |
WO2024181025A1 (fr) * | 2023-03-02 | 2024-09-06 | 本田技研工業株式会社 | Composition |
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IT1271074B (it) * | 1994-11-21 | 1997-05-26 | Italfarmaco Spa | Peptidi ad attivita' antiinfiammatoria |
FR2768335B1 (fr) * | 1997-09-12 | 2000-03-03 | Sederma Sa | Compositions a usage cosmetique ou dermopharmaceutique contenant une association d'extrait d'algue et d'exopolysaccharide |
JP2004203811A (ja) * | 2002-12-26 | 2004-07-22 | Shirako:Kk | 化粧料 |
FR2849596B1 (fr) * | 2003-01-08 | 2005-02-11 | Oreal | Composition pour le traitement d'une peau a tendance acneique |
FR2943685B1 (fr) | 2009-03-25 | 2011-04-29 | Microphyt | Reacteur photosynthetique pour la culture de microorganiques et procede de culture de microorganismes |
CN102712888B (zh) | 2009-11-10 | 2014-05-14 | 米克罗皮公司 | 用于光合反应器的反应罩和相关的光合反应器 |
US9974819B2 (en) * | 2010-10-19 | 2018-05-22 | Cutech S.R.L. | Extracts of microalgae and their application |
MA36538B1 (fr) * | 2013-12-06 | 2016-06-30 | Mascir Moroccan Foundation For Advanced Science Innovation & Res | Utilisation de composition a base d'extraits de microalgues marines pour le traitement d'acne |
BR102014023096A2 (pt) * | 2014-09-12 | 2016-11-16 | Unversidade Fed Do Rio Grande Furg | obtenção de peptídeos com atividade biologica através de hidrólise química e enzimática de proteína de microalga(s) e/ou cianobactéria(s) |
WO2016131824A1 (fr) * | 2015-02-16 | 2016-08-25 | Xantial | Composition de prévention ou de traitement d'un trouble cutané |
FR3047175A1 (fr) * | 2016-01-29 | 2017-08-04 | Expanscience Lab | Compositions cosmetiques, dermatologiques comprenant un extrait de vitex negundo enrichi en polyphenols |
DE102016105029A1 (de) * | 2016-03-18 | 2017-09-21 | Ocean Research & Development Gmbh | Hautaufhellendes Kosmetikum |
-
2018
- 2018-12-28 FR FR1874321A patent/FR3091161B1/fr active Active
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2019
- 2019-12-30 US US17/416,868 patent/US20220071891A1/en active Pending
- 2019-12-30 EP EP19835309.6A patent/EP3902518A1/fr active Pending
- 2019-12-30 JP JP2021537795A patent/JP2022515623A/ja active Pending
- 2019-12-30 KR KR1020217023693A patent/KR20210110336A/ko unknown
- 2019-12-30 WO PCT/EP2019/087171 patent/WO2020136283A1/fr unknown
- 2019-12-30 CN CN201980087677.4A patent/CN113260349B/zh active Active
Also Published As
Publication number | Publication date |
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FR3091161A1 (fr) | 2020-07-03 |
JP2022515623A (ja) | 2022-02-21 |
CN113260349A (zh) | 2021-08-13 |
FR3091161B1 (fr) | 2020-12-11 |
KR20210110336A (ko) | 2021-09-07 |
WO2020136283A1 (fr) | 2020-07-02 |
US20220071891A1 (en) | 2022-03-10 |
CN113260349B (zh) | 2023-06-06 |
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