EP2931231A2 - Kosmetische verwendung eines johannisbrotsamenextrakts als schlankheitswirkstoff - Google Patents

Kosmetische verwendung eines johannisbrotsamenextrakts als schlankheitswirkstoff

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Publication number
EP2931231A2
EP2931231A2 EP13818288.6A EP13818288A EP2931231A2 EP 2931231 A2 EP2931231 A2 EP 2931231A2 EP 13818288 A EP13818288 A EP 13818288A EP 2931231 A2 EP2931231 A2 EP 2931231A2
Authority
EP
European Patent Office
Prior art keywords
extract
vitamin
locust bean
cosmetic use
peptide
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.)
Withdrawn
Application number
EP13818288.6A
Other languages
English (en)
French (fr)
Inventor
Jean-Marie Botto
Nouha Domloge
Frédérique PORTOLAN
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
ISP Investments LLC
Original Assignee
ISP Investments LLC
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Filing date
Publication date
Application filed by ISP Investments LLC filed Critical ISP Investments LLC
Publication of EP2931231A2 publication Critical patent/EP2931231A2/de
Withdrawn legal-status Critical Current

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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
    • 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
    • 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
    • A61K8/96—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
    • A61K8/9706—Algae
    • 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
    • A61K8/96—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
    • A61K8/9728—Fungi, e.g. yeasts
    • 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
    • A61K8/96—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
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • 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
    • A61Q19/06—Preparations for care of the skin for countering cellulitis

Definitions

  • the present invention is in the field of cosmetics and more particularly in the field of cosmetic slimming methods. It relates to the cosmetic use of a carob germ extract (Cemtonia siliqua L.) as a slimming agent. The invention also relates to the cosmetic use of a locust bean extract to increase the expression of aquaglyceroporins and promote the destocking of triglycerides contained in the adipocytes.
  • a carob germ extract Cemtonia siliqua L.
  • the invention also relates to a cosmetic care method comprising the topical application, on at least part of the skin of the body or face, of a carob germ extract (Cemtonia siliqua L.), in a composition comprising a physiologically acceptable medium, to obtain a slimming effect, and more particularly to reduce localized fat overload.
  • a cosmetic care method comprising the topical application, on at least part of the skin of the body or face, of a carob germ extract (Cemtonia siliqua L.), in a composition comprising a physiologically acceptable medium, to obtain a slimming effect, and more particularly to reduce localized fat overload.
  • the subcutaneous adipose tissue is located at the level of the hypodermis. It is a variety of connective tissue in which adipocytes predominate, organized into lobules about 5mm in diameter, separated by fine connective bays. Each adipocyte contains a large lipid vacuole containing essentially triglycerides and whose diameter can vary from 40 to 120 ⁇ .
  • Adipose tissue must be considered as a dynamic reservoir, in constant renewal, ensuring a relay between food intake and the energy needs of the body.
  • adipocytes provide synthesis, storage and release of lipids.
  • Lipid synthesis, or lipogenesis is carried out from triglycerides of food origin and glucose.
  • the triglycerides stored in the adipocytes can be hydrolysed, during lipolysis, to release fatty acids, glycerol and mono- and di-esters of glycerol.
  • the non-esterified fatty acids thus released can either diffuse into the blood and then be available for the energy needs of other cells of the body, or be reused quickly by the adipocyte to generate triglycerides again by lipogenesis.
  • xanthine bases derived from xanthine
  • xanthine such as theophylline, caffeine, theobromine (described in patents FR 2609395, FR 267401), used for their action thus promoting the lipolytic activity of the fat cells.
  • the synthetic peptides such as the Arg-Gly-Ser-NH 2 sequence peptide (described in patent FR 2,858,769), or the sequence peptide Pro-Leu-Asp-Thr-Ala-Lys-Val- Arg-Leu-Gln (described in patent FR 2 879 924) used for its action in the decoupling between the reoxidation of coenzymes and the phosphorylation of ADP in ATP at the level of mitochondria.
  • the synthetic peptides such as the Arg-Gly-Ser-NH 2 sequence peptide (described in patent FR 2,858,769), or the sequence peptide Pro-Leu-Asp-Thr-Ala-Lys-Val- Arg-Leu-Gln (described in patent FR 2 879 924) used for its action in the decoupling between the reoxidation of coenzymes and the phosphorylation of ADP in ATP at the level of mitochondria.
  • Plant extracts such as marine algae extracts of the genus Palmaria or Rhodymenia (described in patent FR 2 887 447), extracts of ginkco biloba (see patent FR 2 669 537), or the flavones or isoflavones of soya (described in WO 01/64177).
  • the solution of the technical problem lies in the cosmetic use of a carob germ extract.
  • the inventors have indeed demonstrated that a carob germ extract acts on the aquaglycéroporines, thus promoting the transport of glycerol released during lipolysis, out of the adipocyte.
  • Aquaporins are a class of transmembrane proteins carrying water and small molecules in solution, between cells and the internal environment. Aquaporins can be classified into two distinct subgroups: aquaporins allowing only the transport of water, and aquaglycéroporines which allow, in addition to the transport of water, the transport of glycerol.
  • aquaglyceroporin 7 has been identified in the membrane of human adipocytes and plays an important role in the metabolism of reserve fats (Mariko Hara-Chikuma et al., Progressive adipocyte hypertrophy in aquaporin-7 deficient mice, J. Biol Chem, vol.280, No. 16, April 22, 2005).
  • aquaglyceroporines therefore make them interesting biological targets for promoting the destocking of lipids contained in adipocytes.
  • the present invention firstly relates to the cosmetic use of a carob germ extract (Ceratonia siliqua L.) as a slimming active agent.
  • active slimming agent in the sense of the present invention, a carob germ extract used to reduce localized fat overload, considered unsightly and often associated with a padded appearance of the skin.
  • the carob seed (plant of the genus Ceratonia), and more particularly the endosperm fraction of this seed, is widely used for its richness in galactomannans, in the food industry under the name "locust bean gum".
  • the seed is the most protein-rich part of the seed and can be easily isolated.
  • peptide extract is intended to mean a mixture of compounds predominantly represented by compounds of a peptide nature, in solution in a large volume of water or other polar solvents or a mixture of these solvents.
  • peptide-like compounds means the protein fragments and the peptides present in the peptide extract according to the invention.
  • topical application is meant the application or spreading of the active agent according to the invention, or a composition containing it, on the surface of the skin or mucosa.
  • physiologically acceptable means that the active agent according to the invention, or a composition containing it, is suitable for coming into contact with the skin or a mucosa without causing toxicity or intolerance reactions.
  • physiologically acceptable any compound suitable for contact with the skin or mucosa without causing toxicity or intolerance reactions.
  • localized adipose overload means an enlarged area of the subcutaneous adipose tissue, which may have an orange peel appearance.
  • active agent and "carob germ extract” will be used interchangeably.
  • the active agent according to the invention can be obtained by extraction of proteins of vegetable origin, followed by controlled hydrolysis which releases the biologically active peptide compounds.
  • peptide extracts and in particular peptide extracts of low molecular weight, has many advantages in cosmetics.
  • hydrolysis and purification make it possible to obtain more stable mixtures, compositions that are more easily reproducible and that do not cause allergic reactions in cosmetics. .
  • locust bean seeds plant of the genus Ceratonia
  • Any method of extraction or purification known to those skilled in the art can be used to prepare the extract according to the invention.
  • the controlled hydrolysis makes it possible to release compounds of peptide nature. It is possible, but not necessary to carry out the invention, either to extract the proteins concerned and then to hydrolyze them, or to perform the hydrolysis on a crude extract and then to purify the compounds of peptidic nature afterwards.
  • the seeds contained in the seeds are crushed in order to obtain a powder or flour.
  • the powder thus obtained can be previously treated with a cellulase to promote the elimination of sugars, and in particular insoluble polysaccharides.
  • the proteins of the seed are then extracted according to the conventional method (modified Osborne, T. B., The Vegetable Proteins, 2nd Edition, Longmans, Green and Co., London, 1924); the locust bean sprout is suspended in an alkaline solution containing an insoluble polyvinylpolypyrrolidone (PVPP) adsorbent product (0.01 - 20%); in fact, it is known that the hydrolyses and the subsequent purifications are facilitated by this means. In particular, the concentration of phenolic-type substances, interacting with proteins, is significantly reduced.
  • the proteins can then be precipitated by varying the ionic strength or acidifying the medium, thereby eliminating soluble components and nucleic acids.
  • the precipitate is then washed with an organic solvent such as, for example, ethanol or methanol, and the solvent is evaporated by drying in vacuo.
  • the protein-rich precipitate is redissolved in water or other solvent and then constitutes a more purified form of the extract.
  • the extraction can also be carried out in neutral or acidic medium always in the presence of polyvinylpolypyrrolidone.
  • the precipitation step is then carried out using a conventional precipitation agent such as salts (sodium chloride, ammonium sulfate) or an organic solvent (alcohol, acetone).
  • a conventional precipitation agent such as salts (sodium chloride, ammonium sulfate) or an organic solvent (alcohol, acetone).
  • the precipitate obtained can be separated from the precipitating agents by dialysis after redissolving in water or another solvent.
  • the soluble fraction containing proteins, carbohydrates and possibly lipids, is collected after centrifugation and filtration steps. This crude solution is then hydrolyzed under mild conditions to generate soluble peptides. Hydrolysis is defined as a chemical reaction involving the cleavage of a molecule by water, this reaction being possible in a neutral, acidic or basic medium. According to the invention, the hydrolysis is carried out chemically and / or advantageously by proteolytic enzymes among which we can then mention endoproteases of plant origin (papain, bromelain, ficin).
  • the hydrolysed carob seed extract obtained at this stage is used.
  • an amount of polyvinylpolypyrrolidone can be added to the reaction medium during this mild hydrolysis step.
  • the resulting extract can be further purified to select low molecular weight peptide compounds.
  • the fractionation can advantageously be carried out by ultrafiltration and / or by a chromatographic type method.
  • a dilution phase is then carried out in water or in any solvent mixture containing water.
  • the active agent according to the invention is advantageously diluted in one or more physiologically acceptable solvents, such as water, glycerol, ethanol, propanediol, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diglycols. , cyclic polyols or any mixture of these solvents.
  • the diluted active agent is then sterilized by ultrafiltration.
  • the locust bean extract is diluted in one or more physiologically acceptable solvents, such as water, glycerol, ethanol, propanediol, butylene glycol or dipropylene glycol. , ethoxylated or propoxylated diglycols, cyclic polyols or any mixture of these solvents.
  • physiologically acceptable solvents such as water, glycerol, ethanol, propanediol, butylene glycol or dipropylene glycol.
  • ethoxylated or propoxylated diglycols cyclic polyols or any mixture of these solvents.
  • a peptide extract is obtained, characterized by a dry weight of 2 to 5 g / kg, a concentration of peptide-like compounds of 1 to 10 g / l, preferably of 1.5 to 3.5 g / l, a concentration of sugars of 0.05 to 1 g / l, preferably of 0.1 to 0.3 g / l and a concentration of polyphenols of less than 1% relative to the dry weight.
  • the carob germ extract has a dry weight of 2.5 g / kg and contains between 1.5 and 3.5 g / l of peptide-like compounds.
  • the extract obtained is composed of peptides with a molecular weight of less than 5 kDa and is characterized by a concentration of sugars of less than 15% and a concentration of polyphenols of less than 1% relative to the dry weight.
  • the locust bean extract is a peptide extract in which the compounds of peptide nature have a molecular weight of less than 5 kDa.
  • the present invention also relates to the use of a locust bean extract to increase the expression of aquaglyceroporins and more particularly of aquaglyceroporin 7.
  • the characteristic molecular activity of the invention is defined in vitro by the ability of the active agent to increase the expression of aquaglyceroporins, either by increasing the protein synthesis of aquaglyceroporines (by direct or indirect modulation of the expression Aquaglyceroporin gene), or by other biological processes such as the stabilization of the aquaglyceroporin protein or the stabilization of messenger RNA transcripts.
  • ⁇ aquaglyceroporin is ⁇ aquaglyceroporin 7.
  • the present invention also relates to the use of a carob germ extract to promote lipolysis, destocking of lipids and the release of glycerol from adipocytes.
  • the biological activity characteristic of the invention is defined in vitro by the ability of the active agent to reduce the size and the number of lipid droplets in the adipocytes.
  • the present invention also relates to a cosmetic care method comprising the topical application on at least part of the skin of the body or face, a carob seed extract (Ceratonia siliqua L.), in a composition comprising a physiologically acceptable medium, to obtain a slimming effect, and more particularly to reduce localized fat overload.
  • a cosmetic care method comprising the topical application on at least part of the skin of the body or face, a carob seed extract (Ceratonia siliqua L.), in a composition comprising a physiologically acceptable medium, to obtain a slimming effect, and more particularly to reduce localized fat overload.
  • the locust bean extract is present at a concentration of between 0.0001% and 20% of the total weight of the composition, and preferably at a concentration of between 0.05% and 5% of the total weight of the composition. in a physiologically acceptable medium.
  • the active agent can be encapsulated or included in a cosmetic vector such as liposomes or any other microcapsule used in the field of cosmetics or adsorbed on powdery organic polymers, mineral supports like talcs and bentonites.
  • compositions for the implementation of the invention may in particular be in the form of an aqueous solution, hydro-alcoholic or oily; an oil-in-water, water-in-oil emulsion or multiple emulsions; they too can be in the form of suspensions, or powders, adapted for application to the skin, mucous membranes, lips and / or hair.
  • compositions may be more or less fluid and have the appearance of a cream, lotion, milk, serum, ointment, gel, paste or paste. a foam. They can also be in solid form, as a stick or be applied to the skin in aerosol form.
  • compositions may furthermore comprise any additive commonly used in the field of application envisaged as well as the adjuvants necessary for their formulation, such as solvents, thickeners, diluents, antioxidants, dyes, sunscreens, self-tanning agents, pigments, fillers, preservatives, perfumes, odor absorbers, cosmetic or pharmaceutical active ingredients, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, etc.
  • adjuvants necessary for their formulation such as solvents, thickeners, diluents, antioxidants, dyes, sunscreens, self-tanning agents, pigments, fillers, preservatives, perfumes, odor absorbers, cosmetic or pharmaceutical active ingredients, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, etc.
  • these adjuvants and their proportions are chosen so as not to adversely affect the desirable properties of the composition according to the invention.
  • These adjuvants may, for example, correspond to 0.01 to 20% of the total weight of the composition.
  • the fatty phase may represent from 5 to 80% by weight and preferably from 5 to 50% by weight relative to the total weight of the composition.
  • the emulsifiers and co-emulsifiers used in the composition will be chosen from those conventionally used in the field under consideration. For example, they can be used in a proportion ranging from 0.3 to 30% by weight, relative to the total weight of the composition.
  • composition that can be used to carry out the invention may comprise, in addition to the active agent according to the invention, at least one other active agent having cosmetic effects similar to and / or complementary to those of the invention.
  • this active agent will be defined as an "additional active agent”.
  • the additional active agent (s) may be chosen from: anti-aging, firming, lightening, moisturizing, draining, microcirculation promoting agents, pharmaceutical agents, exfoliants, desquamants, extracellular matrix stimulating, activating energy metabolism, antibacterials, antifungals , soothing, anti-radical, anti-UV, anti-acne, anti-inflammatory, anesthetic, providing a feeling of warmth, providing a feeling of freshness, slimming.
  • Such additional agents may be chosen from the groups comprising: vitamin A and especially retinoic acid, retinol, retinolpropionate, retinol palmitate,
  • vitamin B3 and more particularly niacinamide, tocopherol niconitate,
  • vitamin B5 vitamin B6, vitamin B12, panthenol
  • vitamin C especially ascorbic acid, ascorbyl glucoside, ascorbyl tetrapalmitate, magnesium and sodium ascorbyl phosphate,
  • amino acids such as arginine, ornithine, hydroxyproline, hydroxyproline dipalmitate, palmitoylglycine, hydroxylysine, methionine and its derivatives, N-acyl amino acid compounds,
  • the natural or synthetic peptides including di-, tri-, tetra-, penta- and hexapeptides and their lipophilic derivatives, isomers and complexed with other species such as a metal ion (eg copper, zinc, manganese) , magnesium, and others).
  • a metal ion eg copper, zinc, manganese
  • MATRKYL ®, ARGIRELINE ®, TM Collaxyl, PEPTIDE VINCI 02 TM, Chronogen TM, LAMINIXYL IS TM, Peptide Q10 TM commercially known as the peptides MATRKYL ®, ARGIRELINE ®, TM Collaxyl, PEPTIDE VINCI 02 TM, Chronogen TM, LAMINIXYL IS TM, Peptide Q10 TM,
  • plant peptide extracts such as extracts of soya, spelled, vine, rapeseed, flax, rice, maize, pea,
  • DHA dehydroacetic acid
  • salicylic acid and its derivatives alpha- and beta-hydroxy acids, silanols,
  • glucosamine D-glucosamine
  • N-acetylglucosamine N-acetyl-D-glucosamine
  • mannosamine N-acetyl mannosamine
  • galactosamine N-acetylgalactosamine
  • extracts of polyphenols such as isoflavones, flavonoids, such as grape extracts, pine extracts, olive extracts,
  • lipids such as ceramides or phospholipids, oils of animal origin, such as squalene or squalane; vegetable oils, such as sweet almond oil, coconut oil, castor oil, jojoba oil, olive oil, rapeseed oil, peanut oil, sunflower oil, wheat germ oil, corn germ oil, soya bean oil, of cotton, alfalfa, poppy, pumpkin, evening primrose, millet, barley, rye, safflower, passionflower, hazelnut, palm, apricot kernel, avocado, calendula ; ethoxylated vegetable oils, shea butter,
  • the invention may comprise at least one additional active agent known for its slimming action, inhibiting lipogenesis or stimulating lipolysis, such as: cyclic and its derivatives, the activating agents of the enzyme adenylate cyclase and the inhibitors of the phosphodiesterase enzyme, centella asiatica extract, asiaticoside and asiatic acid, methyls xanthines, theine, caffeine and its derivatives, theophylline, theobromine, forskolin, esculin and esculoside, ACE inhibitors, Val-Trp peptide, neuropeptide Y inhibitors, enkephalin, gingko biloba extract, dioscorea extract, rutin, yerba mate extract, guarana extract, oligosaccharides, polysaccharides, carnitine, ivy extract, fucus extract, hydrolysed extract of Prunella vulgaris, hydrolysed extract
  • compositions that can be used according to the invention can be applied by any appropriate route, in particular oral or external topical, and the formulation of the compositions will be adapted by those skilled in the art.
  • compositions according to the invention are in a form suitable for topical application.
  • These compositions must therefore contain a physiologically acceptable medium, that is to say compatible with the skin and superficial body growths, and cover all cosmetic forms.
  • the invention is directed to mammals in general, and more particularly to humans.
  • FIG. 1 Immuno-detection of aquaglyceroporin 7 in 3T3-L1 cells.
  • Figure 2 Quantification of the size of the lipid droplets in 3T3-L1 cells.
  • Example 1 Preparation of a Carob Peptide Extract (Ceratonia siliqua L.)
  • the carob germ (Ceratonia siliqua L.) in powder form is dissolved in 70 volumes of water and the pH is adjusted to a value between 4.5 and 5.5.
  • the latter is characterized by a protein content of between 45 and 50% and a sugar content of between 20 and 30%.
  • the dry residue thus obtained is dissolved in 100 volumes of water in the presence of 2% of POLYCLAR® 10.
  • the mixture is adjusted to a pH of between 8.0 and 8.5 with a 2M aqueous sodium hydroxide solution. .
  • a first hydrolysis is carried out using 2% Alcalase® (novozym).
  • the hydrolysis is obtained after 2 hours, with stirring, at 55 ° C.
  • the enzyme is inactivated by heating the solution at 80 ° C for 2 hours. After quenching, the reaction mixture is filtered and the filtrate is collected. It is the intermediate protein extract of locust bean.
  • the peptide and protein compounds of this filtrate are characterized by polyacrylamide gel electrophoresis (NuPAGE® Bis-Tris Pre-cast, Invitrogen gels).
  • the filtrate is heated at 70 ° C for 10 minutes under denaturing reducing conditions in NuPAGE® LDS sample preparation buffer.
  • a solution of antioxidant NuPAGE® is added to the inner vessel (cathode) to prevent the reduced proteins from reoxidizing during electrophoresis.
  • the protein migration is performed in NuPAGE® MES migration buffer in the presence of a molecular weight standard (SeeBlue Plus2). Protein staining is performed using Coomassie® Blue R-250.
  • the protein profile thus obtained shows that the peptide and protein compounds of the filtrate have molecular weights of between 50 and 10 kDa.
  • the carob germ intermediate protein extract is then dissolved in 100 volumes of water in the presence of 2% of POLYCLAR® 10.
  • the mixture is adjusted to a pH of between 4 and 5 with an aqueous solution of hydrochloric acid. 1 Mr.
  • a step of hydrolysis of the proteins is then carried out using an endoprotease.
  • 2% bromelain is added to the reaction medium.
  • Hydrolysis is obtained after 2 hours of stirring at 50 ° C.
  • the enzyme is inactivated by heating the solution at 80 ° C for 2 hours.
  • the carob germ extract is characterized by a dry weight of between 20 and 25 g / kg, a protein content of between 10 and 15 g / l, a sugar content of between 5 and 6 g / 1, an amino acid level of between 1 and 2 g / l and a total polyphenol content of between 0.5 and 1 g / l. Proteins are assayed by a specific colorimetric method (Lowry method)
  • the protein profile of this extract is analyzed by electrophoresis gel. Under the same conditions as previously described, two large families of proteins are observed: the first family, which is a minority, corresponds to proteins of molecular weight of 25 to 20 kDa and the second family, which is very predominant, corresponds to proteins of lower molecular weight. at 5kDa.
  • This extract is then purified by removing proteins of molecular weight greater than 5 kDa by tangential flow filtration steps.
  • the carob germ extract is pumped under pressure through a Pellicon® support equipped with Pellicon® 2 Biomax 30 kDa cassette.
  • the first filtrate obtained is recovered to be filtered again through another Pellicon® 2 Biomax 5 kDa cassette.
  • an orange-yellow carob seed extract brilliant and limpid. It is characterized by a dry weight of between 8 and 9 g / kg, a protein content of between 6 and 7 g / l, a sugar content between 0.3 and 0.5 g / l and a polyphenol content. totals less than 0.1 g / 1.
  • a dilution phase in a water-glycerol mixture in order to obtain a peptide extract characterized by a dry weight of 2 to 5 g / kg, and preferably of 2.5 g / kg, a concentration of compounds of a nature peptide of 1.5 to 3.5 g / l, a sugar concentration of 0.1 to 0.3 g / l (ie less than 15%) and a polyphenol concentration of less than 1%.
  • This purified and diluted extract corresponds to the peptide extract of locust bean germ according to the invention. It is characterized in that the compounds of peptide nature have a molecular weight of less than 5 kDa, that the polyphenol content is less than 1%.
  • the purpose of this study is to determine the influence of the carob extract according to Example 1 on the expression of aquaglyceroporin 7 in differentiated adipocyte cells 3T3-L1.
  • the adipocyte cells 3T3-L1 are cultured in DMEM medium 4.5 g / 1 glucose, 2 mM Glutamine and 10% fetal calf serum.
  • 3T3 cells into adipocytes 2 days after entering the cell confluence phase, the differentiation of 3T3 cells into adipocytes is induced by the addition of a 0.5 mM solution of IBMX, 1 ⁇ l of dexamethasone and 10 ⁇ l of insulin. (Sigma, St. Louis, MO, USA) in the culture medium for 3 days. Then, only the insulin is maintained for 3 to 4 days of additional culture. The cells are then maintained in culture, in standard medium, for a further 3 days.
  • IBMX 0.5 mM solution of IBMX, 1 ⁇ l of dexamethasone and 10 ⁇ l of insulin.
  • the treatment is carried out from the beginning of the induction phase of the differentiation by a daily application for 12 days, either of IX PBS (Lonza, Rockland USA) for the untreated control, or of locust bean extract. dry weight 2.5 g / kg, as obtained in Example 1, diluted to 1% in PBS.
  • a positive control is achieved by a 5-hour treatment with isoproterenol at 10 ⁇ , which is an adrenergic receptor agonist, causing very rapid lipolysis.
  • the cells are washed 3 times with IX PBS (Lonza, Rockland, USA) and fixed with 3.7% formaldehyde (Sigma Aldrich, USA) for 10 min at room temperature.
  • the cell membranes are permeabilized with acetone for 4 min at -20 ° C.
  • Non-specific sites are saturated with 3% bovine serum albumin (Sigma-Aldrich, Steinheim, Germany) for 15 min.
  • the primary antibody (anti-aquaglyceroporin polyclonal rabbit 7, Santa Cruz Biotechnology) diluted at 1/100) is applied for 1.5 hour.
  • Alexa Fluor 488 probe Alexa Fluor 488 donkey anti-rabbit (Invitrogen, Fisher) diluted 1/1000
  • the slides are then mounted in Fluoromount G (Electron Microscopy Science, Hatfield, UK).
  • the cells are examined under the Nikon Eclipse 80i microscope, the 40X objective, and the photos are taken using a Nikon Digital DXM1200C.
  • the carob extract according to Example 1 at 1% causes a significant increase in the expression of aquaglyceroporin 7 in differentiated adipocyte cells 3T3-L1.
  • the purpose of this study is to determine the influence of the carob extract according to Example 1 on the size and the number of lipid droplets contained in differentiated adipocyte cells 3T3-L1.
  • the culture, the differentiation of the 3T3-L1 cells and then the treatment with the test compounds are carried out as in Example 2.
  • Lipid detection is performed using Nile Red fluorescent dye, a phenoxazone that strongly labels intracellular lipids
  • the color of the fluorescence observed is directly dependent on the hydrophobicity of the surrounding medium. This specific property of the Red Nile makes it possible to differentiate the neutral lipids, marked in golden yellow, phospholipids, marked in red.
  • the cells are examined under a fluorescent microscope. Quantification and statistical analysis are performed in the same manner as in Example 2.
  • the intensity of Nile Red labeling, the size and number of the droplets in the differentiated 3T3-L1 cells treated with locust bean extract according to Example 1 at 1% is decreased compared to untreated cells.
  • Quantitative analyzes showed a 35.6% decrease in labeling intensity, a 21.7% decrease in droplet size, and a 29% drop in the number of untreated cells. This is illustrated in Figure 2.
  • the carob extract according to Example 1 at 1% causes a significant decrease in the size and number of lipid droplets in 3T3-L1 differentiated adipocyte cells.
  • phase A The components of phase A are melted at 75 ° C and the components of phase B heated to 75 ° C. Phase A is emulsified at B, then the mixture is cooled below 40 ° C. Phases C and D are then added with constant stirring.
  • phase A and phase B are heated separately at 65 ° C; phase B is incorporated in phase A with stirring.
  • the temperature of the mixture is raised to 83 ° C. and is then cooled to the phase inversion temperature.
  • Phase C is then added.
  • Phase D is incorporated when the temperature reaches less than 40 ° C.
  • the carbopol is dispersed in the water until perfect hydration.
  • the ingredients are then added in the order listed above, with stirring.
  • the mixture is then neutralized with the TEA.
  • the perfume and the dyes are added if necessary.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Birds (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Engineering & Computer Science (AREA)
  • Botany (AREA)
  • Dermatology (AREA)
  • Cosmetics (AREA)
  • Medicines Containing Plant Substances (AREA)
EP13818288.6A 2012-12-11 2013-12-10 Kosmetische verwendung eines johannisbrotsamenextrakts als schlankheitswirkstoff Withdrawn EP2931231A2 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FR1203363A FR2999079B1 (fr) 2012-12-11 2012-12-11 Utilisation cosmetique d'un extrait de germe de caroube en tant qu'agent actif amincissant
PCT/FR2013/053021 WO2014091146A2 (fr) 2012-12-11 2013-12-10 Utilisation cosmetique d'un extrait de germe de caroube en tant qu'agent actif amincissant

Publications (1)

Publication Number Publication Date
EP2931231A2 true EP2931231A2 (de) 2015-10-21

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EP13818288.6A Withdrawn EP2931231A2 (de) 2012-12-11 2013-12-10 Kosmetische verwendung eines johannisbrotsamenextrakts als schlankheitswirkstoff

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Country Link
US (1) US20150297504A1 (de)
EP (1) EP2931231A2 (de)
FR (1) FR2999079B1 (de)
WO (1) WO2014091146A2 (de)

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EP3468530A4 (de) * 2016-06-10 2020-03-11 Clarity Cosmetics Inc. Nichtkomedogene formuliereungen zur haar- und kopfhautpflege und verfahren zur verwendung
CN108484723B (zh) * 2018-01-19 2021-06-11 宁波大学 浒苔来源的血管紧张素转化酶抑制肽及其制备方法和应用
CN115414303A (zh) * 2022-06-29 2022-12-02 广州腾麟生物科技有限公司 一种身体舒悦滋养植物按摩霜
CN117297089A (zh) * 2023-10-11 2023-12-29 广东省农业科学院蚕业与农产品加工研究所 包含多酚提取物和低聚糖提取物的降脂组合物及制备方法

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Publication number Priority date Publication date Assignee Title
LU86934A1 (fr) 1987-06-30 1989-03-08 Oreal Composition cosmetique a action anti-cellulitique et amin-cissante dont le principe actif est une 1-hydroxyalkylxanthine
FR2609395B1 (fr) 1988-02-23 1992-01-10 Serobiologiques Lab Sa Utilisation de bases xanthiques comme principe actif d'une composition pharmaceutique, notamment dermatologique, ou cosmetique, composition notamment utile comme matiere de base pour la preparation de composition pharmaceutique ou cosmetique et composition pharmaceutique ou cosmetique ainsi preparee
FR2669537B1 (fr) 1990-11-28 1993-02-19 Oreal Composition amincissante a base d'alpha-2-bloqueurs.
FR2720604B1 (fr) * 1994-06-03 1996-09-06 Meyhall Chemical Ag Fraction de germe de caroube à haute teneur en protéine.
DE10009424A1 (de) 2000-02-28 2001-09-06 Henkel Kgaa Verwendung von Flavonen oder Isoflavonen zur Cellulite-Behandlung
FR2858769B1 (fr) 2003-08-13 2006-02-10 Soc Extraction Principes Actif Utilisation d'un peptide comme principe actif amincissant
FR2859104A1 (fr) * 2003-08-29 2005-03-04 Saint Laurent Parfums Molecules a activite anti-adipogenique sur cellules adipocytaires humaines
US20050186290A1 (en) * 2003-12-10 2005-08-25 L'oreal Use of aquaglyceroporin modulators as slimming agent
FR2867683B1 (fr) * 2004-03-22 2012-12-21 Lm Cosmetics Compositions cosmetiques ou dermatologiques et leurs applications
FR2879924B1 (fr) 2004-12-23 2007-06-15 Soc Extraction Principes Actif Composition cosmetique amincissante
FR2884418B1 (fr) 2005-04-15 2008-11-21 Soc Extraction Principes Actif Utilisation d'un peptide comme principe actif amincissant
US20080004283A1 (en) * 2005-12-06 2008-01-03 Menarini Ricerche S.P.A. Pharmaceutical Compositions for the Treatment of Cellulite
FR2954697B1 (fr) * 2009-12-24 2016-11-04 Isp Investments Inc Composition cosmetique et/ou pharmaceutique comprenant un extrait de caroube en tant qu'agent actif activateur de l'expression des aquaporines

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Title
None *
See also references of WO2014091146A2 *

Also Published As

Publication number Publication date
WO2014091146A3 (fr) 2014-08-14
WO2014091146A2 (fr) 2014-06-19
FR2999079A1 (fr) 2014-06-13
US20150297504A1 (en) 2015-10-22
FR2999079B1 (fr) 2015-04-24

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