EP0891422A1 - Produit complexe a base de superoxyde dismutase - Google Patents

Produit complexe a base de superoxyde dismutase

Info

Publication number
EP0891422A1
EP0891422A1 EP97919477A EP97919477A EP0891422A1 EP 0891422 A1 EP0891422 A1 EP 0891422A1 EP 97919477 A EP97919477 A EP 97919477A EP 97919477 A EP97919477 A EP 97919477A EP 0891422 A1 EP0891422 A1 EP 0891422A1
Authority
EP
European Patent Office
Prior art keywords
sod
peroxidase
complex
cofactor
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.)
Withdrawn
Application number
EP97919477A
Other languages
German (de)
English (en)
French (fr)
Inventor
Delphine Bresson-Rival
Patrick Boivin
Guy Linden
Eric Perrier
Gérard HUMBERT
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.)
BASF Beauty Care Solutions France SAS
Original Assignee
Coletica SA
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.)
Filing date
Publication date
Application filed by Coletica SA filed Critical Coletica SA
Publication of EP0891422A1 publication Critical patent/EP0891422A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
    • A23L11/30—Removing undesirable substances, e.g. bitter substances
    • A23L11/33—Removing undesirable substances, e.g. bitter substances using enzymes; Enzymatic transformation of pulses or legumes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/06—Enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
    • A23L5/25—Removal of unwanted matter, e.g. deodorisation or detoxification using enzymes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/44—Oxidoreductases (1)
    • A61K38/446—Superoxide dismutase (1.15)
    • 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
    • A61K8/60—Sugars; Derivatives 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
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/66—Enzymes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K8/67—Vitamins
    • A61K8/676—Ascorbic acid, i.e. vitamin C
    • 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
    • A61K8/67—Vitamins
    • A61K8/678—Tocopherol, i.e. vitamin E
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • 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
    • A61Q19/08—Anti-ageing preparations
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0065—Oxidoreductases (1.) acting on hydrogen peroxide as acceptor (1.11)
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0089—Oxidoreductases (1.) acting on superoxide as acceptor (1.15)

Definitions

  • the present invention relates to a complex product based on SOD, and cosmetic, pharmaceutical or agrifood compositions containing it, and to an extraction process.
  • the present invention also relates to a process for extracting / purifying a superoxide dismutase from plant seeds after germination, and to its combination with an H2O2 trapping agent, in particular a peroxidase, preferably accompanied by its enzymatic cofactor.
  • This enzymatic complex is very stable and can be used in compositions, in particular cosmetic, pharmaceutical or agrifood compositions.
  • the invention aims to obtain a superoxide dismutase
  • SOD styrene dioxide
  • H2O2 hydrogen peroxide
  • the invention thus finds an advantageous application in cosmetic compositions, pharmaceutical or dermatological compositions, or agrifood compositions.
  • free radicals are atoms or molecules that have an unpaired electron on their external orbital. These are extremely unstable compounds that can react with the most stable molecules to pair their electrons.
  • Oxygenated free radicals are constantly formed in the body, in the mitochondrics or during phagocytosis processes. Physical factors such as exposure to ultraviolet light as well as the environment in which humans evolve are also factors of high production of free radicals at the level of biological compounds. These factors are, for example, automobile pollution, tobacco, ionizing radiation, etc. Oxygenated free radicals are formed by partial reduction of molecular oxygen. The capture of an electron by oxygen, will generate the radical O2 " 1" superoxide. This anion is not very reactive but it can generate very reactive radical species.
  • free radicals Due to their high reactivity, free radicals are capable of attacking all cellular constituents and of causing serious alterations: in the skin, free radicals, in particular the superoxide anion, have as major target the collagen but also elastin fibers, glycosaminoglycans and proteoglycans, intracellular DNA, phospholipids from cell membranes, etc.
  • Collagen and elastic fibers directly capture the free radicals formed, at the cost of various degradations: breaking of chains and release of small peptides degraded by non-specific proteases, interchain bonds reducing elasticity.
  • compositions based on antioxidants in all fields, and in particular in cosmetics, pharmacy or agrifood.
  • cosmetics such compositions have been put on the market and are called anti-aging or anti-aging products.
  • antioxidants currently used in cosmetics or in pharmacies are either chemical molecules that collect free radicals, or enzyme systems, the major drawback of which is very poor stability even at room temperature.
  • the lipophilic sensors used are generally vitamin E and ⁇ -carotene, components of cell membranes which protect them from lipid peroxidation by reducing the peroxide radicals formed.
  • the hydrophilic antioxidants used generally consist of vitamin C, which acts in aqueous media with respect to super ⁇ and hydroxyl radicals, but also by enzymatic cofactors such as glutathione and zinc.
  • Glutathione is the cofactor of many enzymes involved in anti-radical defense (such as glutathione peroxidase, glutathione transferase, etc.) while zinc is the cofactor of copper-zinc SOD.
  • SOD The enzyme used in cosmetics or pharmacy is SOD, the role of which is to ensure the almost instantaneous destruction of superoxide radicals which are potentially harmful to the body and the tissues of the skin.
  • the reaction catalyzed by SOD is the disproportionation of the superoxide anion into hydrogen peroxide or hydrogen peroxide according to the following chemical reaction:
  • the Cu-Zn SOD present in the cutaneous tissues, constitutes the first line of enzymatic defense against the free radicals generated by the ultraviolet rays.
  • the enzyme sees its detoxification potential decrease sharply after irradiation with ultraviolet rays. This observation has, moreover, led professionals in cosmetology or pharmacy to incorporate SOD into formulations in order to reduce the level of free radicals present in the skin tissues.
  • the main object of the present invention is to solve the new technical problem consisting in providing a solution which makes it possible to obtain SOD of plant origin in large quantities with a very good yield so as to use it on an industrial scale, in particular in the cosmetic, pharmaceutical or food industry.
  • the present invention also aims to solve the new technical problem consisting in the supply of a solution which makes it possible to obtain an SOD of plant origin which is extremely active and also extremely stable, preferably by being capable of retaining 80% of activity for 1 month (35 days) at room temperature and an activity of the order of 50% at 45 ° C., thus allowing effective incorporation into anti-free radical compositions, in particular in cosmetic, pharmaceutical or agrifood compositions.
  • the present invention relates to a complex product based on superoxide dismutase or SOD stabilized by an H2O2 scavenger, advantageously a peroxidase, preferably vegetable, with more preferably a peroxidase cofactor also called a specific reducing substrate. peroxidase.
  • the SOD is of vegetable origin.
  • the SOD was obtained by extraction of plant seeds after germination.
  • the plant seeds are cereal seeds or seeds of leguminous plants or seeds of oleaginous plants.
  • grains or seeds are equivalent.
  • cereals all cereals can be used, in particular rye, corn, wheat or barley, preferably barley, among the varieties of barley that can be used, spring or winter varieties can be used .
  • legumes seeds of any legume plant will be used, but preferably lentil or pea seeds.
  • oilseeds seeds of any oilseed plant will be used, but preferably soybeans.
  • the above-mentioned germination is controlled, it is carried out in aqueous atmosphere or moist medium, preferably in the presence of an agent promoting the germination at a temperature between 4 ° C and 50 ° C, advantageously at room temperature or cold for one or more days.
  • H2O2 is sufficient to trap I ⁇ 2O2 formed by the SOD used, when the H2O2 trapping agent is a peroxidase, the amount of peroxidase is between approximately 0.01 and approximately 1 peroxidase unit per SOD unit, advantageously between
  • the SOD / H2O2 trapping agent complex in particular a peroxidase
  • an enzymatic cofactor in particular an enzymatic cofactor of peroxidase, present at a concentration of between 0.001M and 1M.
  • the aforementioned peroxidase is chosen from the group consisting of a horseradish peroxidase, a lactoperoxidase, a glutathione peroxidase or a spinal cord peroxidase or myelo peroxy ⁇ dase, and the specific reducing substrate of peroxidase is preferably chosen from glutathione, phenol, guaiacol, pyrogallol, mesitol, 3,5-dichloro-2-hydroxybenzene sulfonic acid (DCHBS), aniline, p-toluidine, o-phenylene diamine, mesidine, ascorbic acid, dihydroxymaleic acid, cytochrome C, iodides, uric acid, phenolphthalein, 2,2'-azido-di (3-ethylbcnzo-thiazoline (6) sulfonic acid (ABTS) and compounds such as SCN-, Cl " , Br- or I-.
  • ABTS
  • the H2O2 trapping agent is a vegetable peroxidase, preferably extracted from the black radish.
  • the above-mentioned peroxidase cofactor is chosen from the group consisting of uric acid, ascorbic acid or dihydroxymaleic acid.
  • the SOD / H2O2 trapping agent complex in particular peroxidase, with optionally a peroxidase cofactor, is further stabilized by at least one sugar, in particular a monosaccharide or a disaccharide, and / or at least one polyol in particular having a molecular weight of between 50 and 1000 g / mole, in particular at a concentration of between 10 and 50% by weight of SOD complex, in the form of crude extract or in purified form.
  • the complex product also comprises an antioxidant advantageously lipophilic, preferably from the tocopherol family and their derivatives, in particular their esters, such as acetates, linoleates or even phosphates, in an effective antioxidant amount.
  • an antioxidant advantageously lipophilic, preferably from the tocopherol family and their derivatives, in particular their esters, such as acetates, linoleates or even phosphates, in an effective antioxidant amount.
  • the present invention also relates to the use of the SOD-based complex as defined above as one of the principles or active ingredients of a composition in particular with anti-free radical activity, for example a cosmetic, pharmaceutical or food industry.
  • the present invention also relates to a composition in particular with anti-free radical activity, for example cosmetic, pharmaceutical or food composition, characterized in that it comprises as one of the active ingredients a complex product based on superoxide dismutase or SOD stabilized by an H2O2 trapping agent, advantageously a peroxidase, preferably vegetable, with more preferably a peroxidase cofactor also called a specific reducing substrate for peroxidase.
  • the proportion of SOD is between 0.01 and 10% by weight of the weight of the total composition.
  • the proportion of incorporation is variable and will depend on the application envisaged. Generally, the proportion of incorporation of SOD will be from 0.01 to 30% by weight, better from 0.1 to 10% by weight and even better still of the order of 1 to 5%.
  • the peroxydascs catalyze the destruction of oxygenated water (H2O2) produced during the disproportionation reaction catalyzed by the SOD of the superoxide anion in oxygenated water as recalled at the beginning of the description in the presence of a specific reducing substrate known as a peroxidase cofactor. .
  • Peroxidases are numerous and well known to those skilled in the art. They are currently extracted from the spinal cord or myelo peroxidase, milk or lactoperoxidase, bovine or possibly human erythrocytes or glutathione peroxidase or even preferably according to the invention from black radish or horseradish peroxidase.
  • the specific reducing substrates for peroxidases are well known to those skilled in the art and are preferably chosen from glutathione, phenol, guaiacol, pyrogallol, mcsitol, 3,5-dichloro-2-hydroxybenzene sulfonic acid (DCHBS ), aniline, p-toluidine, o-phenylene diamine, mesidine, ascorbic acid, dihydroxymaleic acid, cytochrome C, iodides, uric acid, phenolphtaleinc, 2,2'-azido-di (3-ethylbcnzo-thiazoline (6) sulfonic acid ) (ABTS) and compounds such as SCN-, Cl-, Br- or I-.
  • an SOD complex with a vegetable peroxidase preferably extracted from black radish, advantageously accompanied by its cofactor, preferably consisting of uric acid, ascorbic acid or dihydroxymaleic acid.
  • the amount of peroxidase added to the SOD is a function of the rate of hydrogen peroxide released by the SOD, therefore of its enzymatic activity.
  • the number of peroxidase units to be added to the SOD solution is between approximately
  • the peroxidase enzymatic cofactor as previously described which may preferably be uric acid, is added to the enzymatic complex at a concentration of between 0.001 M and 1 M.
  • the SOD or the SOD / peroxidase / peroxidase cofactor complex can be further stabilized by at least one sugar and / or at least one polyol added at a concentration of between 10 and 50% by weight of SOD or of the final complex, in the form of crude extract or in purified form.
  • sugar use will be made in particular of a monosaccharide or a disaccharide, and in particular trehalose, and as polyol, use in particular of a polyol having an average molecular weight of between 50 and 1000 g / mol, for example glycerol, sorbitol, maltitol and mannitol.
  • a sugar or a polyol allows the activity of SOD to be stabilized in a particularly unexpected manner.
  • an advantageously lipophilic antioxidant is added, preferably from the tocophcrol family and their derivatives, such as their esters such as acetates, linoleates or even phosphates since it has been unexpectedly discovered that such an antioxidant provides increased stability to SOD.
  • the tocopherol phosphates are, for example, described in US patent N'5,387,579 from LVMH RECHERCHE. This result is particularly unexpected since the stability of the SOD enzyme is not linked to oxidation problems and therefore the mechanism of action is currently unknown. Since this antioxidant is advantageously lipophilic, it will be added to the composition in an oily phase, that is to say in general in the context of the formation of an emulsion.
  • the concentration of antioxidant agent will generally be from 0.01 to 3%, preferably from 0.1 to 1%, by weight relative to the total weight of the final composition.
  • the present invention also relates to a method for extracting superoxide dismutase or SOD, characterized in that the seeds of plants having undergone a germination stage are used controlled so as to obtain a maximum SOD activity during germination and to extract the SOD activity at the time when this SOD activity is maximum by bringing the seeds of plants into contact with a humid aqueous atmosphere or medium during one or more days at room temperature, and in that a step of extracting the SOD from the germinated seeds is carried out comprising grinding the germinated seed followed by extraction in an aqueous solution at a temperature of between 4 ° C. and 50 ° C, preferably at room temperature, for a period of time sufficient to carry out the extraction of SOD, generally a few tens of minutes to one or more hours, followed by filtering and recovering the filtrate containing SOD.
  • the germination step comprises suspending the seeds of plants in an aqueous solution for several days at room temperature.
  • this germination step takes place in the presence of an agent promoting germination, again preferably consisting of a compound belonging to the family of gibbercllincs.
  • the germination step is preceded by a step of quenching in an aqueous solution at room temperature or cold for one or more days, for example two days.
  • the step of extracting the SOD from the germinated seeds comprises grinding the germinated seed followed by extraction in a buffered aqueous solution having a pH close to 8 at room temperature for a period of time sufficient to carry out the extraction of the SOD, generally from a few tens of minutes to one or more hours, followed by filtration and recovery of the filtrate containing the SOD.
  • further purification of the SOD can be carried out by treatment of the filtrate with a precipitation agent making it possible to remove the undesirable proteins including the proteases or even the lipoxygenase, and recovery of the fraction not precipitated or constituting the supernatant which contains SOD.
  • a membrane preferably having a cutoff threshold of between 6000 and 8000 Dalton.
  • the method according to the invention it is possible to carry out a stabilization of SOD obtained directly after the abovementioned extraction step, that is to say before further purification, or after purification, by addition of an H2O2 scavenger, such as a peroxidase, preferably with a peroxidase cofactor also known as a specific peroxidase reducing substrate.
  • an H2O2 scavenger such as a peroxidase
  • a peroxidase cofactor also known as a specific peroxidase reducing substrate.
  • the amount of peroxidase added to the SOD expressed in a ratio of peroxidase unit relative to the SOD units is between 10 and 400 according to the invention whereas for the enzymatic cofactor of peroxidase it is preferably present in a concentration between 0.001 M and 1 M.
  • the SOD by adding after the extraction step, or after the purification step, at least one sugar, in particular a monosaccharide or a disaccharide and / or at least one polyol, in particular a polyol having an average molecular weight of between 50 and 1000 g / mole.
  • the sugar or the polyol will be added at a concentration of between 10 and 50% by weight of the SOD or of the final SOD complex, in the form of a crude or purified extract.
  • This Nebot method is based on the property that any catalyst provided with an SOD activity of accelerating, at alkaline pH, the autoxidation of a reagent RI into a chromophorc absorbing visible light, according to the following reaction:
  • the SOD-525 method also uses a second reagent R2 which makes it possible to eliminate major interference due to the mercaptans which could be present in the sample to be analyzed, such as for example glutathione, using an alkylation reaction. very fast, depending on the reaction:
  • the SOD activity is measured at a pH of 8.8, which allows optimal sensitivity of the assay without inactivation of known natural SODs, such as, for example, copper-zinc, manganese or iron SODs.
  • This kit includes two reagents, RI and R2, and a buffer solution, buffer 3, which are respectively:
  • RI a solution of chromogenic reagent RI in 3.2.10 _ - M HC1 of the above formula
  • reaction speed is determined by evaluating the maximum slope of the curve obtained. This slope corresponds to an RI autoxidation phase. The results are expressed in absorbance units per minute.
  • Vc and Vs be the reaction rates corresponding to the control and to the sample respectively.
  • SOD activity of the sample to be analyzed is determined by calculating the correspondence between the experimental ratio Vs / Vc and the SOD activity, deduced from the following equation:
  • One SOD-525 activity unit defined by Oxis corresponds to a Vs / Vc ratio equal to 2 under the above conditions.
  • the value obtained is then multiplied by the dilution factor of the sample (factor 25) in the assay procedure.
  • the results are expressed in units of SOD-525 activity per ml of sample.
  • the SOD enzymatic activity is measured after dilution of the SOD solution obtained in order to remain within values of Vs / Vc ratio making it possible to establish a correct correspondence with the SOD activity.
  • the SOD solution obtained will generally be diluted, depending on the degree of purification resulting from the extraction process, up to a factor of 300, in order to obtain a Vs / Vc ratio of between 1 and 2.
  • the activity of the SOD solution is determined by taking into account the dilution factor of the sample of the assay procedure (factor 25) and the dilution factor of the SOD solution itself (for example up to 'at a factor of 300).
  • compositions in particular with free anti-radical activity for example cosmetic, pharmaceutical or food compositions, characterized in that they comprise, as one of the active ingredients, a vegetable SOD obtained from plant seeds having undergone a stage germination.
  • a vegetable SOD obtained from plant seeds having undergone a stage germination.
  • the extraction protocol from germinated barley is as follows: After germination of the barley grains, the malt is ground, then extracted into an aqueous buffer solution having a pH of the order of 8, which is preferably constituted by Tris HC1 50mM + EDTA ImM, for approximately 1 h at room temperature.
  • the extract is then filtered to remove the bark, then centrifuged at 4000 G for 20 minutes to remove the polysaccharide part and the filtrate or supernatant is recovered on which the SOD activity is determined by the aforementioned method of Nebot C, et al. . (1993) using a commercial kit from Oxis International, 94385 BONNEUIL / MARNE, FRANCE.
  • An SOD-525 activity / g of starting dry matter is obtained.
  • the SOD activity is measured after dilution of 1 ml of the SOD extract in 14 ml of water. 40 ⁇ l of the diluted solution are dosed.
  • the measured Vs / Vc ratio is 1.95.
  • the SOD activity is 0.95 SOD-525 units.
  • the filtrate or supernatant obtained at the end of the above centrifugation is subjected to precipitation by a precipitating agent eliminating the undesirable proteins such as the proteases or also the lipoxygenase, for example ammonium sulfate at a concentration of 390 g / 1.
  • Centrifugation is carried out at 4000 G for 20 minutes at 20 ° C to remove the precipitate and the liquid fraction consisting of the supernatant which contains the SOD is recovered.
  • the supernatant can advantageously be dialyzed to eliminate small molecules and salts present in high concentration having a molecular weight of less than 6000 Dalton. This dialysis is carried out against demineralized water with membranes whose cutoff threshold is 6000-
  • SOD has a molecular weight of the order of 30,000, which does not pass into pigs with such membranes, which makes it possible to increase the specific activity of the enzyme.
  • the SOD activity measured on the dialysate according to the preceding method is 7,500 SOD-525 units / g of starting dry matter.
  • step c) From the dialysate obtained in step c) above, it is still possible to carry out a much more thorough purification by working on a chromatographic column.
  • Example 2 The same protocol is used as that described in Example 1 and the SOD activity on the extraction product from step b is measured and 116 IUso ⁇ Vg of starting dry matter are obtained.
  • Example 2 The same protocol is used as that described in Example 1 and the SOD activity on the product obtained in step b is measured. We obtain a SOD activity of 80 UIsoD ⁇ S ⁇ c starting dry matter.
  • the SOD activity was measured from germinated barley of the Dallas variety by varying the germination time and by respecting the other general conditions of the germination protocol of example 1-a. The results indicated in Table I below are obtained.
  • Stabilization of SOD with a polvol SOD in the form of a crude extract obtained in Example 1 step b) retains 60% of its enzymatic activity after 46 days at 20 ° C.
  • the stability of the SOD enzyme is unexpectedly improved since 76% of the initial activity is found after 46 days at 20 ° C.
  • SOD by adding a sugar, in particular a monosaccharide or a disaccharide.
  • Example 8 If one proceeds as described in Example 6, but adding 30% by weight of trehalose to the crude SOD extract obtained in Example 1 step b), relative to the weight of the final solution, that 75% of initial activity is retained after 46 days at 20 ° C., which constitutes a remarkable improvement in stability.
  • Example 8 If one proceeds as described in Example 6, but adding 30% by weight of trehalose to the crude SOD extract obtained in Example 1 step b), relative to the weight of the final solution, that 75% of initial activity is retained after 46 days at 20 ° C., which constitutes a remarkable improvement in stability.
  • the solution concentrated in SOD obtained in step 1-d or obtained in a previous step of Example 1 is complexed or combined with another enzyme capable of destroying the hydrogen peroxide formed.
  • the enzymes which can be used in this sense can be a catalase which transforms H2O2 into H2O and O2 but the catalase is not used insofar as it is included in the list of products not authorized in cosmetology.
  • the lists of peroxidases and cofactors have been given in the introduction to the description.
  • HRP horseradish peroxidase
  • SOD is approximately 10 peroxidase units per 400 SOD units, the SOD unit being measured according to the method previously described and the peroxidase unit being determined by the method described by Bergmcyer H.U. in Methods of
  • the peroxidase enzyme cofactor constituted here by uric acid is added to the SOD / pcroxydase enzyme complex in a concentration of between 0.01 and 1 M, and preferably 0.5 M.
  • the complex thus formed is still stabilized by the addition of sugar or polyol added at a concentration chosen in this example to 30% by weight of the final solution.
  • This SOD / pcroxydasc / cofactcur complex can be used as it is for formulating compositions with anti-free radical or free radical scavenging activity, whether cosmetic, pharmaceutical or agrifood or other compositions.
  • This complex is also the subject of tests of stability, anti-free radical capacity and toxicology which are the subject of Examples 9, 10 and 11 respectively below.
  • Example 8 The stability of the enzymatic activity of the complex formed in Example 8 was carried out at 20 ° C and 45 ° C respectively and the results are listed in Table IV below.
  • the first method uses an enzymatic system constituted by xanthine oxidase which, by oxidizing its substrate, xanthine, generates superoxide radicals.
  • the latter are capable of reducing cytochrome C (Fe ⁇ + ) to cytochrome C (Fe * - + ), a reaction the kinetics of which can be followed by UV-visible spectrophotometric at 550 nm. They are also the radical substrates SODs which can thus compete with cytochrome C for the capture and elimination of superoxide radicals. By adding a preparation containing SOD activity, this reduction kinetics is slowed down. The SOD activity is calculated by calculating the percentage of inhibition of the reduction of cytochrome C by the SOD tested.
  • SOD unit expressed in the International System (1 UIJJQD) corresponds to the activity required to reduce by 50% the rate of reduction of cytochrome C, at a temperature of 25 ° C and pH 7.8.
  • test sample is diluted so as to determine the conditions for which 50% inhibition of the reduction of cytochrome C is obtained, corresponding to 1 international system unit.
  • the total SOD activity of the test sample is then determined by reducing the volume of sample dosed to the total volume of the sample and taking into account the initial dilution factor of the sample.
  • the SOD activity of the complex formed in Example 8 determined by this method is 4455 UISOD units / " 1 ⁇
  • the second method consists of a dosing kit marketed by the company OXIS INTERNATIONAL, under the trade name kit SOD-525, based on the property that any catalyst provided with an SOD activity, of accelerating autoxidation of a tetracyclic catechol derivative (5, 6, 6a, 1 lb-tetrahydro-3, 9, 10-trihydroxybenzofluorènc).
  • This assay method makes it possible to measure the SOD activity in the form of a SOD-525 unit which is defined as the quantity of dismutase which multiplies by 2 the oxidation rate of the fluorinated derivative.
  • the SOD activity of the complex formed in Example 8, measured by this method, is 4375 SOD-525 units / ml
  • the third method used is the Paramagnetic Resonance of the electron or abbreviated as RPE.
  • EPR is a technique used to characterize the spin states of single electrons in molecules, more particularly used to characterize free radicals. Evaluation of the anti-radical activity of the complex
  • the spin trap used is 5,5-dimethyl-1-pyrroline-1-oxide (DMPO).
  • DMPO 5,5-dimethyl-1-pyrroline-1-oxide
  • the superoxide anion is produced by the xanthine enzyme system
  • Example 8 The SOD / pcroxydasc / cofactor complex obtained in Example 8 is dissolved in ultrapure water at various concentrations.
  • the hydroxyl radical is produced by photolysis (UVB) of 0.2% (v / v) hydrogen peroxide in aqueous solution, in the presence of DMPO (160 mM).
  • the SOD / pcroxydasc / cofactor complex obtained in Example 8 is dissolved in ultrapure water at various concentrations.
  • the intensities of the EPR signal are calculated by double integration of the lower field line.
  • the protection percentages of the test product are obtained from the values of the controls without the test product.
  • the SOD / peroxidase / cofactor complex obtained in Example 8 decreases in a dose-dependent manner the RPE signal of the superoxide anion. At 5 and 10% (v / v), the complex inhibits the RPE signal at 44% and 54%.
  • the SOD / peroxidase / cofactor complex obtained in Example 8 decreases in a dose-dependent manner the RPE signal of the hydroxyl radical. At a concentration of 0.3% (v / v), the effect is spectacular since it inhibits the RPE signal by 93%.
  • the SOD / peroxidase / cofactor complex obtained in Example 8 exhibits very interesting anti-free radical effects with respect to the superoxide anion and the hydroxyl radical even at low concentrations, generally used in cosmetic formulations.
  • the free radicals formed in a culture of human fibroblasts are quantified by an appropriate method.
  • the use of anti-free radical agents makes it possible to reduce the oxidative stress, and the calculation of the effectiveness of the plant SOD / pcroxydase / cofactor complex according to Example 8 at different concentrations is carried out.
  • Normal human fibroblasts are incubated for 1 hour in the presence of the vegetable SOD / pcroxydase / cofactor complex according to Example 8, tested at 0.01, 1 and 10% (v / v) in demineralized water.
  • the cells are subjected to radical stress caused by UVA light radiation (10 J / cm ⁇ ).
  • the free radicals formed (hydroperoxides) are detected using an appropriate probe, which transforms in the presence of hydroperoxides, into a quantifiable fluorescent derivative.
  • E (%) [(FTI-FI) / (FI-FNI)] x (-100) where: E: Efficiency in percentage
  • FNI Fluorescence observed with Non-Irradiated Fibroblasts
  • FI Fluorescence observed with Irradiated Fibroblasts
  • An aqueous solution containing 1% of plant SOD / peroxidase / cofactor complex formed according to the invention is capable of reducing by more than 80% the harmful effects linked to oxidative stress generated by UVA.
  • the free radicals formed following UVA irradiation peroxidate the unsaturated lipids of the skin tissues. Stripping allows the corneocytes to be removed with the peroxidized entities. In the reaction medium, the peroxides are revealed using a fluorescent probe which emits a fluorescence directly proportional to the level of peroxides present.
  • An aqueous solution containing 1% of vegetable SOD complex / peroxidase / cofactor formed according to the invention is applied 4 times at 2 hour intervals (2 ⁇ l / cm ⁇ ).
  • the horny layer is removed by 2 successive stripping.
  • An aqueous solution containing 1% of plant SOD / peroxidase / cofactor complex formed according to the invention is capable of protecting, with an efficiency of 65%, the cutaneous radical stress induced by UVA irradiation.
  • Toxicology Toxicology tests were carried out on the complex according to the invention obtained in Example 8 on the evaluation of primary skin irritation in rabbits (9a), on the evaluation of ocular irritation in rabbits (9b), on the absence of abnormal toxicity by single oral administration in the rat (9c) and by the study of the sensitizing power on the guinea pig (9d).
  • Example 8 The preparation obtained in Example 8 was applied without dilution at a dose of 0.5 ml to the skin of 3 rabbits according to the method recommended by the directive.
  • Example 8 11b- Evolution of eye irritation in rabbits
  • the same preparation obtained in Example 8 was instilled pure all at once, at a rate of 0.1 ml, into the eyes of 3 rabbits according to the method recommended by the OECD directive 405 of 24 February 1987 on the study of "acute irritant / corrosive effect on the eyes".
  • Example 8 The results of this test make it possible to conclude that the preparation obtained in Example 8 according to the invention can be considered to be non-irritating to the eyes within the meaning of Directive 91/326 EEC used pure or without dilution. l ie- Test on the absence of abnormal toxicity by single oral administration in rats
  • Example 8 The preparation obtained in Example 8 according to the invention was administered once orally at a dose of 5 g / kg of body weight, to 5 male rats and 5 female rats according to a protocol inspired by the directive of OECD No. 401 of February 24, 1987 and adapted to cosmetic products.
  • the SOD complex solution obtained in Example 8 was the subject of a search for sensitizing power according to the method of Magnusson and Kligman published in J. invest. Dcrm. (1969), 52, pages 268-276.
  • This SOD complex solution was applied as such to the skin of 35 guinea pigs previously treated with Freund's adjuvant and divided into 2 experimental batches, respectively control batch and treated batch.
  • Example 8 The preparation obtained in Example 8 according to the invention is incorporated into a cream at a concentration of 5% by weight relative to the total weight of this cream.
  • This cream has the following composition:
  • Vegetable SOD / peroxidase / cofactcur according to the invention of Example 8 .... 5% The enzymatic activity in the cream is measured after separation of the aqueous and oily phases, carried out under specific conditions: 2 g of cream are diluted 5 times in 8 g of 0.2 M Tris (hydroxymethyl) aminomethane buffer, pH 8.5, then 10% NaCl are added. The mixture is subjected to violent stirring (of the type obtained with an Ultra-turax) for 5 minutes and then centrifuged at 5000 rpm for 25 minutes. The enzymatic activity is measured directly in the extracted aqueous phase.
  • Example 8 The stability of the enzymatic activity of the preparation obtained in Example 8 incorporated into a cream, whose description is given above, has been studied at 20 ° C and 45 ° C for 40 days. The results are listed in Table V.
  • the enzymatic activity found in the cream after 40 days at 20 ° C is 60% for the stabilized SOD form. Even more remarkable, the enzymatic activity found in the cream stored for 40 days at 45 # C is 50%, while the non-stabilized form does not withstand temperature.
  • Complex stability studies according to the invention, incorporated in a cream, show that the complex has a remarkable stability at 20 ° C and particularly unexpected # 45 C.
  • the SOD solution obtained in step le of Example 1 is used, in which a number of units of horseradish vegetable peroxidase, known as
  • HRP in a ratio of HRP units to SOD units between 10 and
  • the vegetable SOD / horseradish hydroxydase preparation thus obtained can be used as such or preferably combined with an enzyme cofactor, for example uric acid or ascorbic acid or dihydroxymaleic acid.
  • an enzyme cofactor for example uric acid or ascorbic acid or dihydroxymaleic acid.
  • Plant SOD complex / pcroxydasc of Arthromvces ramosus The procedure is as in Example 13A except that a peroxidase obtained from commercially available Arthromyccs ramosus is used, for example under the reference Sigma P 4794.
  • Example 14 The procedure is as described in Example 13A except that a peroxidase extracted from the soybean commercially available under the reference Sigma P 1462 is used.
  • Example 14
  • the stabilization of the vegetable SOD / peroxidase / cofactor enzyme complex obtained in Example 8 is improved by adding a sugar or polyol at least according to the following variant embodiments:
  • Example 14A Addition of Glvercerol To the preparation obtained in Example 8 according to the invention, the glycerol is added at a concentration chosen in this example to 30% by weight of the final solution, with stirring at room temperature.
  • Example 14A The procedure is as in Example 14A, except that sorbitol is used.
  • Example 14A The procedure is as in Example 14A, except that mannitol is used.
  • Example 15 The procedure is as in Example 14A, except that trehalose is used.
  • Example 8 The procedure is as described in Example 8, except that peroxidase cofactors other than the uric acid which was used in Example 8 are used.
  • Example 8 The procedure is as described in Example 8, except that ascorbic acid is used in place of uric acid at a concentration of between 0.01 and 1 M, and preferably 0, 5 M.
  • Example 8 The procedure is as described in Example 8, except that dihydroxymaleic acid is used in place of uric acid, at a concentration of between 0.01 and 1 M, and preferably 0 , 5 M.
  • Cosmetic composition of tvpc oil-in-water emulsion usable more particularly for anti-free radical cosmetic preparations. for antiaging purposes. anti-wrinkle and anti-stress of the skin
  • a composition is prepared in the form of an oil-in-water emulsion in a conventional manner from the following five fractions A, B, C, D and E having the percentage composition indicated:
  • A- Oily phase comprising:
  • phase B is mixed in phase A, then phase C, phase D and again phase E until an oil-in-water emulsion is obtained.
  • composition with anti-inflammatory activity comprises
  • This pharmaceutical composition comprises, in addition to a conventional pharmaceutical active principle, a preparation as obtained in Example 8 according to the invention, in an amount of 5% by weight, in mixture with a pharmaceutically acceptable excipient.
  • This agrifood composition has stability against rancidity and comprises, in addition to the conventional agrifood active principles, 5% by weight of preparation obtained in Example 8 according to the invention which are incorporated at the same time as the other active ingredients. .

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Epidemiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Birds (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Nutrition Science (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • Botany (AREA)
  • Gerontology & Geriatric Medicine (AREA)
  • Agronomy & Crop Science (AREA)
  • Dermatology (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Pain & Pain Management (AREA)
  • Rheumatology (AREA)
  • Enzymes And Modification Thereof (AREA)
EP97919477A 1996-04-03 1997-04-03 Produit complexe a base de superoxyde dismutase Withdrawn EP0891422A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
FR9604165 1996-04-03
FR9604165A FR2747044B1 (fr) 1996-04-03 1996-04-03 Utilisation de graines de vegetaux apres germination comme source de superoxyde dismutase et compositions cosmetiques, pharmaceutiques ou agroalimentaires contenant une telle superoxyde dismutase vegetale, et procede d'extraction
PCT/FR1997/000603 WO1997038095A1 (fr) 1996-04-03 1997-04-03 Produit complexe a base de superoxyde dismutase

Publications (1)

Publication Number Publication Date
EP0891422A1 true EP0891422A1 (fr) 1999-01-20

Family

ID=9490855

Family Applications (1)

Application Number Title Priority Date Filing Date
EP97919477A Withdrawn EP0891422A1 (fr) 1996-04-03 1997-04-03 Produit complexe a base de superoxyde dismutase

Country Status (6)

Country Link
US (1) US5904921A (enExample)
EP (1) EP0891422A1 (enExample)
JP (1) JP2000508530A (enExample)
KR (1) KR20000005196A (enExample)
FR (1) FR2747044B1 (enExample)
WO (1) WO1997038095A1 (enExample)

Families Citing this family (26)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE19653736C2 (de) * 1996-12-12 2002-11-21 Lancaster Group Gmbh Kosmetisches Präparat mit Peptidzusatz
TW466116B (en) 1997-03-04 2001-12-01 Hayashibara Biochem Lab Reduction inhibitory agent for active-oxygen eliminating activity, method for inhibiting the reduction of said activity, and composition containing said agent
FR2777785B1 (fr) * 1998-04-27 2001-03-30 Sederma Sa Utilisation de l'association peroxydase de raifort/acide cafeique dans des compositions cosmetiques ou dermopharma- ceutiques destinees a prevenir et/ou corriger les degats cutanes induits par les formes radicalaires de l'oxygene
US6309656B1 (en) * 1998-11-27 2001-10-30 Peter T. Pugliese Cosmetic and skin protective compositions
EP1262167A1 (de) * 2001-06-01 2002-12-04 Cognis France S.A. Kosmetische Zubereitungen enthaltend ein Extrakt von keimenden Pflanzen
JP4565311B2 (ja) * 2002-10-03 2010-10-20 東洋紡績株式会社 酵素の安定化方法および組成物
US7491509B2 (en) 2003-02-03 2009-02-17 Fraunhofer Usa, Inc. System for expression of genes in plants
US7683238B2 (en) * 2002-11-12 2010-03-23 iBio, Inc. and Fraunhofer USA, Inc. Production of pharmaceutically active proteins in sprouted seedlings
CN1313606C (zh) * 2003-07-23 2007-05-02 韩伯钧 一种从玉米中提取sod酶精粉的制造方法
NZ566918A (en) * 2005-08-26 2011-09-30 Protech Res Pty Ltd Process for enhancing hydrolase enzyme production in a grain by wounding the grain prior to germination
US7687650B2 (en) 2006-02-03 2010-03-30 Jr Chem, Llc Chemical compositions and methods of making them
US7927614B2 (en) * 2006-02-03 2011-04-19 Jr Chem, Llc Anti-aging treatment using copper and zinc compositions
US7897800B2 (en) 2006-02-03 2011-03-01 Jr Chem, Llc Chemical compositions and methods of making them
FR2908998A1 (fr) * 2006-11-24 2008-05-30 Bao Quoc Ho Nouveau produit enzymatique et son utilisation.
JP2008194030A (ja) * 2007-01-15 2008-08-28 Kitakyushu Foundation For The Advancement Of Industry Science & Technology 異常プリオンタンパク質検出用組成物ならびにそれを用いた検出方法、異常プリオンタンパク質増殖阻止方法、そのために使用できる異常プリオンタンパク質増殖阻止用飼料及び食品、およびスーパーオキシド生成防止方法およびプリオン由来ペプチドもしくはプリオンタンパク質検出用組成物ならびに検出方法
US8273791B2 (en) 2008-01-04 2012-09-25 Jr Chem, Llc Compositions, kits and regimens for the treatment of skin, especially décolletage
US20160184354A1 (en) 2009-01-23 2016-06-30 Jr Chem, Llc Rosacea treatments and kits for performing them
US8952057B2 (en) 2011-01-11 2015-02-10 Jr Chem, Llc Compositions for anorectal use and methods for treating anorectal disorders
FR3031040B1 (fr) 2014-12-29 2018-09-07 Universite De Bretagne Occidentale Procede d'obtention d'un extrait proteique concentre en superoxydase dismutase (sod)
JP2015147798A (ja) * 2015-04-20 2015-08-20 ビーエーエスエフ ビューティ ケア ソリューションズ フランス エスエーエス 弾性繊維の形成を刺激するための、ｌｏｘｌ（リシルオキシダーゼ類似）アイソフォームの合成及び活性の刺激
FR3072031B1 (fr) * 2017-10-11 2019-10-04 Alaena Cosmetiques Principe actif cosmetique a base d'extraits de graines germees
FR3094612B1 (fr) 2019-04-05 2022-08-19 Laboratoires Iphym Composition comprenant un extrait de cafe vert et une Superoxyde dismutase (SOD)
CN111920803B (zh) * 2020-09-29 2021-07-27 中国农业大学 用于提高超氧化物歧化酶的活性和/或热稳定性的制剂及其应用
CN113373122B (zh) * 2021-07-29 2025-03-07 辽宁未来生物科技有限公司 一种新型玉米超氧化物歧化酶多酶的制备方法
US20250195624A1 (en) * 2022-02-21 2025-06-19 Isocell S.A.S. Use of superoxide dismutase for the treatment and/or prevention of allergic asthma
KR20250071617A (ko) * 2023-11-15 2025-05-22 (주)바이오제닉스 단백질의 활성을 유지하기 위한 안정화 복합체 조성물 및 이의 제조방법

Family Cites Families (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3637640A (en) * 1970-05-04 1972-01-25 Diagnostic Data Inc Orgotein stabilized with saccharide process and products
US3773929A (en) * 1972-11-02 1973-11-20 Diagnostic Data Inc Pharmaceutical compositions comprising orgotein and their use
FR2225443B1 (enExample) * 1973-04-16 1976-11-12 Anvar
DE2462631C2 (de) * 1973-04-16 1980-12-04 Agence Nationale De Valorisation De La Recherche (Anvar), Neuilly-Sur-Seine, Hauts-De-Seine (Frankreich) Superoxiddismutase, Verfahren zu deren Herstellung und deren Verwendung
JPS5587712A (en) * 1978-12-26 1980-07-02 Yoshihide Hagiwara Skin cosmetic
IL77817A (en) * 1986-02-06 1995-06-29 Yeda Res & Dev Process for controlling plant growth and herbicidal compositions therefor
FR2611495A1 (fr) * 1987-02-24 1988-09-09 Cochand Georges Produit cosmetologique destine a etre applique sur la peau
FR2656874B1 (fr) * 1990-01-11 1992-04-03 Commissariat Energie Atomique Procede de production et d'extraction d'anti-oxydants a partir d'une culture de micro-organismes et photobioreacteur pour la mise en óoeuvre de ce procede.
FR2657526B1 (fr) * 1990-01-31 1994-10-28 Lvmh Rech Utilisation d'un phosphate d'alpha-tocopherol, ou de l'un de ses derives, pour la preparation de compositions cosmetiques, dermatologiques, ou pharmaceutiques; compositions ainsi obtenues.
JP3008131B2 (ja) * 1990-11-14 2000-02-14 ロレアル グリセリンから誘導される非イオン両親媒性化合物、その調製方法、相応する中間体化合物及び前記化合物を含有する組成物
FR2675997B1 (fr) * 1991-05-03 1993-12-24 Oreal Composition topique anti radicaux libres a base de superoxyde-dismutase et d'un derive phosphonique.
JPH06199694A (ja) * 1992-03-06 1994-07-19 Yunie:Kk 血圧安定化治療剤
FR2693208A1 (fr) * 1992-07-02 1994-01-07 Inocosm Laboratoires Procédé d'obtention d'une composition enzymatique d'origine végétale ainsi que composition obtenue par la mise en Óoeuvre de ce procédé.
RU2046140C1 (ru) * 1992-07-08 1995-10-20 Акционерное общество закрытого типа - Химико-биологическое объединение при РАН Способ выделения ферментного комплекса из культивируемых растительных клеток
US5843641A (en) * 1993-02-26 1998-12-01 Massachusetts Institute Of Technology Methods for the daignosis, of familial amyotrophic lateral sclerosis
FR2706465B1 (fr) * 1993-06-11 1995-08-25 Heliosynthese Sa Procédé de production et d'extraction de superoxyde-dismutases thermostables à partir d'une culture de micro-organismes photosynthétiques.
JP3836891B2 (ja) * 1994-12-22 2006-10-25 リアル化学株式会社 頭髪用化粧料および毛染め毛髪の処理方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9738095A1 *

Also Published As

Publication number Publication date
US5904921A (en) 1999-05-18
JP2000508530A (ja) 2000-07-11
WO1997038095A1 (fr) 1997-10-16
KR20000005196A (ko) 2000-01-25
FR2747044A1 (fr) 1997-10-10
FR2747044B1 (fr) 1998-06-26

Similar Documents

Publication Publication Date Title
EP0891422A1 (fr) Produit complexe a base de superoxyde dismutase
Yokota et al. Scavenging of reactive oxygen species by Eriobotrya japonica seed extract
EP0504236B1 (fr) Utilisation d'extraits d'algues pour la preparation de compositions pharmaceutiques, cosmetiques, alimentaires ou a usage agricole
EP2018149B1 (fr) Principe actif cosmetique compose de ferrulate d'arginine et d'un extrait de microalgue et ses utilisations
EP1243586B1 (fr) Fraction phénolique riche en phloridzine et son utilisation en tant qu'agent cosmétique, alimentaire ou nutraceutique.
EP1820492B1 (fr) Composition protectrice et régénérante
EP2421884B1 (fr) Hydrolysats peptidiques activateurs du proteasome et compositions les contenant
EP0684944B1 (fr) Produit de couplage de l'histamine et d'un acide amine
WO2014167247A2 (fr) Extrait d'arthrobacter agilis pour son utilisation notamment en cosmetique
JP3502415B2 (ja) メイラード反応阻害剤
JP3643038B2 (ja) メイラード反応阻害剤、ヒスタミン遊離抑制剤、活性酸素生成抑制剤及び過酸化脂質生成抑制剤
CA2487693A1 (en) Method for producing preparations rich in tocotrienol
CN116196235B (zh) 一种抗氧化组合物及其制备方法、化妆品和应用
EP0582147A2 (fr) Composition cosmétique ou dermatologique anti-uréase
EP2419439A1 (fr) Composition cosmétique et/ou pharmaceutique comprenant un hydrolysat peptidique apaisant
FR2940113A1 (fr) Hydrolysat de gomme de caroube, son procede de preparation et son utilisation en cosmetique capillaire
WO2008145853A2 (fr) Utlisation d'un principe actif issu de l'amarante (amaranthus) pour preparer une composition destinee a activer l'energie cellulaire et a proteger la peau des dommages oxydatifs
EP3638203A1 (fr) Compositions cosmetiques comprenant des extraits naturels et leurs utilisations
FR3134515A1 (fr) Extraits de fleurs de Crocus sativus, compositions les comprenant et leurs utilisations dans les soins buccaux
CH711956A2 (fr) Composition cosmétique ayant des propriétés anti-radicalaires.
JP3199309B2 (ja) 化粧料
EP1778162A1 (fr) Utilisation d'un extrait de triticum monococcum comme principe actif dans une composition cosmetique et/ou pharmaceutique
FR2690343A1 (fr) Composition anti-radicalaire et anti-lipoperoxydante.
FR2943534A1 (fr) Utilisation cosmetique anti-radicalaire d'un principe actif issu de myrtus communis, notamment de tanins hydrolysables d'un principe actif issu de myrtus communis.
FR2855749A1 (fr) Procede d'obtention d'un principe actif dote de proprietes de photoprotection, principe actif obtenu et compositions l'incluant

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 19981001

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): CH DE ES FR GB LI

17Q First examination report despatched

Effective date: 20030701

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20040112