EP3240565A1 - Procede d'obtention d'un extrait proteique concentre en superoxyde dismutase (sod) - Google Patents
Procede d'obtention d'un extrait proteique concentre en superoxyde dismutase (sod)Info
- Publication number
- EP3240565A1 EP3240565A1 EP15817894.7A EP15817894A EP3240565A1 EP 3240565 A1 EP3240565 A1 EP 3240565A1 EP 15817894 A EP15817894 A EP 15817894A EP 3240565 A1 EP3240565 A1 EP 3240565A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- sod
- limonium
- plant
- protein
- protein extract
- 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- Withdrawn
Links
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y115/00—Oxidoreductases acting on superoxide as acceptor (1.15)
- C12Y115/01—Oxidoreductases acting on superoxide as acceptor (1.15) with NAD or NADP as acceptor (1.15.1)
- C12Y115/01001—Superoxide dismutase (1.15.1.1)
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C19/00—Cheese; Cheese preparations; Making thereof
- A23C19/02—Making cheese curd
- A23C19/04—Making cheese curd characterised by the use of specific enzymes of vegetable or animal origin
- A23C19/043—Enzymes other than proteolytic enzymes or milk clotting enzymes, e.g. lipase, lysosyme
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C3/00—Preservation of milk or milk preparations
- A23C3/08—Preservation of milk or milk preparations by addition of preservatives
- A23C3/085—Inorganic compounds, e.g. lactoperoxidase - H2O2 systems
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/70—Fixation, conservation, or encapsulation of flavouring agents
- A23L27/72—Encapsulation
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3571—Microorganisms; Enzymes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/185—Vegetable proteins
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- 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
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/645—Proteins of vegetable origin; Derivatives or degradation products thereof
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- A—HUMAN NECESSITIES
- 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
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- 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]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
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- 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]
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- 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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/52—Stabilizers
- A61K2800/522—Antioxidants; Radical scavengers
Definitions
- the present invention relates to a process for obtaining a concentrated protein extract superoxide dismutase (SOD) and the extract obtained by this method and its use.
- SOD superoxide dismutase
- the present invention also relates to a composition comprising the extract obtained by the method of the invention.
- SOD Superoxide dismutases
- ROS reactive oxygen species
- SOD is therefore the first line of defense against these EORs, which are strongly implicated in various neurodegenerative diseases (Alzheimer's disease, Parkinson), atherosclerosis, rheumatoid arthritis, Crohn's disease and some cancers (Campana et al., 2004; Valdivia et al., 2009).
- SODs are metalloproteins that contain in their active site Cu / Zn, Mn or Fe atoms. These metals play an important role in the mechanism of action of SOD. For example, for Cu / Zn SOD, Cu 2+ is first reduced by the superoxide radical (0 2 " ), which is thus detoxified into oxygen.The reduced copper then yields an electron to a new superoxide molecule which in the presence of two protons, generates H 2 0 2 , much less toxic than 0 2 ' " .
- SODs Due to their antioxidant capacity, SODs are widely used in different fields:
- SODs protect the skin and hair by maintaining the integrity of the natural keratin structure (FR 7331354).
- This dietary supplement is particularly appropriate to fight against the overproduction of free radicals, especially when the natural defenses of the body are weakened: elderly subjects, sun exposure, smoking, stress, intense physical exercise, etc. It can prevent certain chronic conditions involving oxidative stress or slow their evolution, thus improving the patient's living conditions.
- plant extracts rich in SOD are incorporated in foods for sport animals such as racehorses or racing pigeons to increase their resistance to intense efforts.
- the same extracts are incorporated into feed before and during the transport of farmed fish and crustaceans (shrimp, fry, trout, salmon, etc.) to reduce the mortality rate during transport.
- extracts rich in SOD are introduced into the diet of females of dairy farms (cows, goats, etc.).
- Extracts rich in SOD are used in some food formulations to improve the growth performance of chickens and turkeys, animals often faced with poorly controlled digestive imbalances.
- SOD can be of very different origins: animal (for example bovine or porcine), human (for example placental or sanguine), microbial (for example bacteria or yeasts), etc.
- SODs are of plant origin (fungi, seaweeds, spinach, cereals, fruits and vegetables, etc.).
- SOD is a fragile protein, it is necessary to have processes that make it possible to obtain it in large quantities or better still, in a smaller quantity but with a high specific activity, without increasing the use of chemical compounds capable of alter the functionality of this protein and decrease its specific activity.
- SOD can be obtained simply by protein extraction from plants comprising it in large quantities and having a high specific activity. It is preferable that such SOD extraction processes do not involve the use of a multitude of solvents, especially organic solvents, which require the implementation of additional purification steps and degrade part of the SOD. .
- the extracts obtained must be clean, that is to say devoid of the chemical reagents that were necessarily used during the extraction process, which will optimize the use of these products. extracts especially in food and feed. It is also desirable that the extract obtained by these methods contain other beneficial active substances, naturally present in plants, such as polyphenols which have antioxidant properties.
- the present invention therefore aims to implement a SOD extraction process having a very high specific activity.
- This process is simple, fast, inexpensive; it makes it possible to obtain an extract devoid of chemical reagents, it contains other natural substances having a beneficial effect and thus overcomes the disadvantages of the SOD extraction methods known in the prior art.
- the inventors have surprisingly discovered that the protein extracts obtained from the species of a family of littoral plants are so rich in SOD activity that a simple aqueous extraction from the crushed plant, without the addition of other solvents (for example for example, alcoholic solvents) and without additional purification steps, is sufficient to obtain extracts where the specific activity of SOD is at least 700 IU / mg of protein. This level greatly exceeds the highest level (126 IU / mg) reported so far in the literature for a variety of melon (Vouldoukis et al., 2004) considered as a reference among natural sources of SOD.
- the inventors have implemented a simple process for extracting SOD, the cost of which is low and which makes it possible to obtain very active protein extracts in SOD devoid of chemical reagents frequently used in conventional extraction processes and which contains other natural substances having a beneficial effect, thus overcoming the disadvantages of SOD plant extraction processes of the prior art.
- a first subject of the present invention therefore relates to a process for obtaining a concentrated superoxide dismutase (SOD) protein extract having a specific activity of at least 700 IU / mg of protein comprising a single protein extraction step from of a plant selected from any of the species of the Plombaginaceae family.
- SOD superoxide dismutase
- extraction process means the implementation of a set of steps for obtaining a protein extract from a source, preferably a plant source. said steps comprising at least one step of recovering a liquid phase after contacting (under appropriate conditions) the solvent with said source, and then a step of purification comprising a precipitation step, in order to obtain the desired protein extract.
- the expression "in a single extraction step” means that a single step is carried out among the steps mentioned above, namely the recovery of the liquid phase after contacting. solvent with the source, preferably the vegetable source.
- the protein extraction method according to the present invention therefore does not include SOD protein purification steps and / or SOD protein enrichment steps.
- the term “specific activity” refers to the SOD activity related to the amount of protein, that is to say the activity of the SOD in the protein extract.
- the inventors have discovered that, on average, the extracts obtained directly (without purification and / or enrichment steps) by a simple step of contacting the water with crushed plants of the family Plombaginaceae, family of plants commonly represented in the littoral environment, have a specific activity of at least 700 IU / mg of protein.
- this specific activity may be at least 1000 IU / mg, more particularly at least 1200 IU / mg and even more particularly at least 1400 IU / mg.
- IU designates the enzyme unit of SOD in the international system.
- the enzymatic unit of SOD is defined by Me Cord and Fridovitch (1969).
- catalytic activity refers to the SOD activity reported to the source plant biomass.
- This catalytic activity is, in the species of the family Plombaginaceae, in a range between 3000 and 14000 IU / g MS (activity units / g of material dry), particularly between 5000 and 13000 IU / g MS and even more particularly between 7000 and 12000 IU / g MS.
- the inventors have also carried out comparative tests (see comparative examples) which demonstrate that the species of the Apiaceae, Aizoaceae, Brassicaceae and Chenopodiaceae families exhibit a SOD catalytic activity of between 120 and 1250 IU / g MS, which is much lower than that of species of the family Plombaginaceae.
- the Plombaginaceae family is a cosmopolitan family of dicotyledonous plants.
- the species of this family are present everywhere, particularly in saline and cold environments where their adaptability gives them a selective advantage.
- the Plombaginaceae family is represented by the three genera Plumbago, Limonium and Armeria. Among them, the genera Limonium and Armeria are the most frequently encountered of this family. They are found on the entire coastline, from the Channel to the Mediterranean, via the Atlantic coasts.
- the process of the invention is carried out from species of the genera Limonium and Armeria found throughout the French coast, preferably on the French Atlantic coast.
- the genus Armeria contains only one species which is named Armeria maritima.
- the genus Limonium includes perennial herbaceous plants with purple flowers that grow on sandy or poor soils of the Atlantic coast and in salt marshes.
- the species of the genus Armeria are perennial plants of the dunes or littoral rocks, whose flowering lasts from July to September.
- the extraction of SOD is carried out from the plants selected from the group consisting of the species Limonium latifolium, Limonium normannicum, Limonium vulgare, Limonium tunetanum, Limonium densiflorum, Limonium Pruinosum, Limonium delicatulum, Limonium spathulatum, Limonium boitardii, Limonium wrightii and Armeria maritima.
- the SOD extraction process is carried out from the species Limonium latifolium which is easily cultivable or accessible in the region where the present invention has been implemented.
- Limonium latifolium or sea lavender (also called statice) is a perennial herb that grows on sandy or poor soils of the Atlantic coast and in salt marshes. This sea lavender has antioxidant properties due to its phenolic compounds and contains coloring agents (flowers and roots).
- Limonium latifolium is much richer in SOD specific activity than the plant species previously exploited in the agri-food industry as sources of SOD. This means that it is not necessary to have a large quantity of the plant to obtain a very enzymatically active SOD, hence a lower cost of producing a finished product enriched with SOD.
- the species L. latifolium is an easily cultivable species, with a faster development cycle than melon, without known predator, hence the possibility of producing this plant under conditions corresponding to the regulations on organic crops.
- Limonium latifolium is a halophytic salt-resistant species
- salt stress is known to stimulate the synthesis of antioxidant compounds including enzymes (Jithesh et al., 2006, Ben Hamed et al. Li et al., 2008, Hameed et al., 2014)
- the inventors have thus carried out additional tests which demonstrate that the SOD activity can be increased by 10 to 20% in plants of the species L. latifolium grown in the presence of salt at a concentration of 20 g / L NaCl (ie 2/3 of the salt concentration in seawater).
- the inventors have demonstrated that the species Limonium latifolium has particularly advantageous properties and advantages as described above.
- the inventors have also demonstrated that other species of the genus Limonium have these same properties and advantages and can thus be easily cultivated under controlled conditions in the presence of salt in order to increase the specific activity of SOD.
- the extraction is carried out from the aerial parts of the plant, preferably from the leaves which are the fleshier organs and the richest in SOD.
- the method of the invention comprises a single extraction step consisting in the recovery of an aqueous phase containing the plant extract after contacting the crushed plant of the family Plombaginaceae with water only.
- the inventors have demonstrated that plants of the family Plombaginaceae are so rich in SOD that the simple aqueous extraction without addition of other solvents or chemical substances makes it possible to obtain an extract whose specific SOD activity is from minus 700 IU / mg of protein.
- the extraction is carried out in the presence of water or an aqueous buffer of the phosphate buffer type, therefore in the absence of toxic organic solvents of the alcohol type (eg: ethanol, methanol, propanol, isopropanol, butanol), aromatic solvents (eg pyridine, toluene, chlorobenzene or xylene), carbonyl solvents (eg acetone, dimethyl formamide or methyl pyrrolidone), esters (eg: ethyl acetate), sulfoxides (eg dimethyl sulfoxide), nitriles (eg acetonitrile), halogenated solvents (eg dichloromethane, dichloroethane, chloroform, carbon tetrachloride, chlorobenzene), ether-oxides (eg diethylene glycol, tetrahydrofuran, ethyl ether).
- the alcohol type eg: ethanol, methanol, prop
- glycerin is added, which acts as a stabilizer.
- said plant prior to the extraction step, said plant can be washed, cut, crushed, micronized and sieved.
- the extraction can be carried out from the plant material previously frozen or dried so as to allow its storage and / or storage.
- the drying of the plant is carried out by lyophilization in order to avoid degradation of the enzyme.
- a second subject of the present invention relates to an SOD concentrated plant protein extract having a specific activity of at least 700 IU / mg proteins, the extract being obtained by the method of the invention as defined above.
- the extract obtained according to the method of the invention has all or part of the characteristics of said process as described below.
- the extract obtained by the process of the invention is an extract which does not contain chemical reagents usually used in conventional extraction processes and the elimination of which is frequently linked to the reduction in the quality of the extract since it causes the decline of the specific activity of SOD and the destruction of certain substances naturally present in the plant and having a beneficial effect.
- the extract of the invention can be kept cold, that is to say, at a temperature of between 4 ° C and -30 ° C, preferably between -2 ° C and -30 ° C, more particularly between -18 ° C and -27 ° C.
- the preservation of the extract can be carried out by drying it, preferably drying by freeze-drying.
- the extract of the present invention may comprise other substances naturally present in the family of Plombaginaceae species having a beneficial effect, such as polyphenols which are known for their antioxidant properties.
- the extract of the invention mainly contains the SOD Cu / Zn isoform.
- the extract of the present invention has the advantage of being very rich in SOD activity as well as in other antioxidant substances, such as polyphenols, and because of its process for obtaining, not to contain or to contain very few chemical reagents usually used during extraction.
- the extract of the invention is therefore perfectly suitable for use in medicine and / or in human and / or animal cosmetology.
- a third subject of the present invention thus relates to a cosmetic or pharmaceutical composition
- a cosmetic or pharmaceutical composition comprising the extract obtained according to the process of the present invention and a cosmetically or pharmaceutically acceptable vehicle, respectively.
- the term "pharmaceutically acceptable carrier” means any material suitable for use in a pharmaceutical product.
- a pharmaceutically acceptable vehicle mention may be made of lactose, optionally modified starch, cellulose, hydroxypropylmethylcellulose, mannitol, sorbitol, xylitol, dextrose, calcium sulphate, calcium phosphate, calcium lactate, dextrates, inositol, calcium carbonate, glycine, bentonite and mixtures thereof.
- the pharmaceutically acceptable vehicles are chosen from the group consisting of prolamines, that is to say reserve proteins accumulated by the plants of the Poaceae family such as cereals.
- compositions and the medicaments according to the invention may be in various forms, in particular in a form chosen from the group consisting of injectable solutions, tablets, capsules, dragees, syrups, suspensions, solutions, powders, granules, emulsions, microspheres.
- sustained release dosage forms such as capsules, or optionally coated tablets for sustained release will be preferred.
- the pharmaceutical composition and the medicaments according to the invention are administrable by different routes.
- administration routes that can be used for the pharmaceutical composition and the medicinal products according to the invention, mention may be made of the oral route, the rectal route, the cutaneous route, the nasal route, the sublingual route, the parenteral route, especially intradermal route. subcutaneous, intramuscular, intravenous, intraarterial, intra-articular, intra-pleural and intraperitoneal.
- the pharmaceutical composition and the medicaments according to the invention can be administered in one or more times or in continuous release.
- the pharmaceutical composition or the medicaments according to the invention are administered in continuous release, such as in the form of an infusion or by means of a pharmaceutical form, such as an optionally coated capsule or tablet, sustained and / or delayed release.
- the term "cosmetically acceptable vehicle” means a vehicle adapted for use in contact with human and animal cells, in particular the cells of the epidermis, without toxicity, irritation, allergic response and proportionate to a reasonable benefit / risk ratio.
- the cosmetic composition according to the invention may comprise one or more formulation agents or additives of known and conventional use in cosmetic and dermatological compositions such as, by way of examples and in a nonlimiting manner, softeners, colorants, film-forming active agents, surfactants, perfumes, preservatives, emulsifiers, oils, glycols, vitamins such as vitamin E, UV filters, etc.
- formulation agents or additives such as, by way of examples and in a nonlimiting manner, softeners, colorants, film-forming active agents, surfactants, perfumes, preservatives, emulsifiers, oils, glycols, vitamins such as vitamin E, UV filters, etc.
- the cosmetic composition according to the invention may be in any form known to those skilled in the field of cosmetology and dermatology with no other pharmaceutical restriction than the application on the face and body.
- the compositions according to the invention are in the form of a gel, a cream, a lotion, an oil, a milk, a spray, etc.
- the extract of the invention can be obtained by aqueous extraction without the use of other chemical reagents (alcohols, organic solvents, buffers), it is particularly suitable for use in food and feed.
- the present invention further relates to a food or nutraceutical composition
- a food or nutraceutical composition comprising a protein extract obtained according to the method of the invention and a food additive or a nutraceutically acceptable vehicle, respectively.
- the food additives may be chosen from all the food additives well known to those skilled in the art, for example, in the group comprising preservatives, food colorants, antioxidants, lactates, citrates, orthophosphates, malates, adipates, alginates, gums, di- and triphosphates, etc.
- the term "nutraceutical composition” is intended to mean a composition relating to a product made from food and marketed in the form of a tablet, powder or potion having a physiological beneficial effect against chronic diseases.
- nutraceutically acceptable carrier in the sense of the present invention, any material that is suitable for use in a nutraceutical product.
- a nutraceutically acceptable carrier mention may be made of dicalcium phosphate, calcium alginate, magnesium carbonate, magnesium stearate, silicon dioxide, calcium chloride, and the like.
- the nutraceutical composition according to the invention is administrable especially orally, for example, in the form of breakable tablets or not, film-coated or not, granules, capsules, capsules, or in the form of free powders preferably packaged in single sachets. , or compressed powder.
- the extract obtained by the process of the invention is intended for use in a food and / or nutraceutical composition
- said extract comprises of the order of 50000 to 200000 IU / g of solids, preferably of the order of 100000 to 150000 IU / g, or, by weight, approximately 1 to 4% preferably 2 to 3% of superoxide dismutase in the dry extract.
- dry extract is intended to mean the dry residue which remains after desiccation of an extract and, for example, by vacuum evaporation at 40 ° C. or by freeze-drying.
- composition according to the invention may comprise from 0.1% to 20%, more particularly from 0.1% to 10% or from 0.5% to 5%, in particular from 1 to 3% of superoxide dismutase. by weight relative to the total weight of the composition.
- the pharmaceutical composition according to the invention comprises from 5% to 20%, particularly from 5% to 15% and more particularly from 5% to 10% of superoxide dismutase by weight relative to the total weight of the composition. .
- the food composition and / or the nutraceutical composition according to the invention may comprise from 0.1% to 5%, particularly from 0.5% to 3% and more particularly from 1 to 2% of superoxide. dismutase by weight relative to the total weight of the composition.
- a fourth aspect of the present invention relates to a cosmetic use of a protein extract as defined according to the invention as an antioxidant cosmetic agent, in particular for combating aging of the skin and / or integuments, against the rays. UV, etc.
- a fifth aspect of the invention relates to the use of a SOD protein extract as defined above for the preparation of cheeses and / or the preservation of dairy products.
- SOD can couple to catalase and in this way effectively inhibit the anarchic oxidation of lipids in dairy products, allowing prolonged storage of milk.
- a sixth aspect of the present invention relates to the SOD protein extract as defined above as a medicament.
- the invention relates to a SOD protein extract according to the invention for its use in the prevention and / or treatment of diseases selected from the group comprising:
- psychiatric diseases preferably schizophrenia
- - neurodegenerative diseases preferably Alzheimer 's disease or Parkinson' s disease
- cardiovascular diseases preferably myocardial infarction or atherosclerosis
- inflammatory diseases preferably Crohn's disease or rheumatoid arthritis
- - chronic diseases preferably diabetes
- cancerous diseases preferably selected from the group consisting of skin cancer, breast cancer, esophageal cancer, stomach cancer, liver cancer, colon cancer and lung cancer .
- Species tested are usually found on sand dunes, rocks or salt marshes along the Breton coastline.
- the species tested are Limonium normannicum, L. vulgare, L. latifolium, L. tunetanum, L. densiflorum, L. pruinosum, L. delicatulum, L. spathulatum and Armeria maritima.
- plants of the species L. latifolium were cultivated under the following controlled conditions: Photoperiod: 8h / 16h (night / day); Temperature: 14/23 ° C (night / day); Relative humidity: 50-70%; Salinity: 0, 5, 10, 20 g / L NaCl.
- Extraction of SOD proteins is carried out immediately after the leaves are taken from the mother plants or after preservation at -80 ° C. For this, the leaves are ground in water or a 100 mM pH phosphate buffer. 7.8 for 15 minutes at 4 ° C. After filtration or centrifugation at 14,000 g for 30 minutes at 4 ° C., the supernatant is collected (liquid phase). The soluble protein content as well as the enzymatic activities are determined from the supernatant.
- the soluble protein content is determined in the supernatant of each enzymatic extract according to the method of Bradford (1976). 1.4 Measurement of SOD activities
- the substrate of SOD is very unstable and has a very short life span (10 "9 s), so the assay methods are very indirect.
- the superoxide radicals produced by this system reduce the nitroblue tetrazolium (NBT) to formazan blue.Two sets of tubes were prepared The former are controls, maintained in the dark and containing a mixture in 50 mM phosphate buffer pH 7.8 composed of 0.1 mM EDTA, methionine (13 mM), NBT (75 ⁇ l), riboflavin (2
- the second series of tubes is intended for the determination of the SOD activity and these tubes contain the same reaction mixture but they are maintained for 15 minutes under a lighting of 15 W.
- the measurement of the absorbance is carried out at 560 nm.
- SOD nzymatic correspond
- the catalytic activity of the enzyme is then related to the mass of tissues that have been extracted while the specific activity is related to the amount of protein in the extract.
- Table 1 Results of measurements of catalytic activity and specific activity of SOD in Plombaginaceae species.
- Table 2 Comparative test results of catalytic activity and specific activity of SOD in species of different families of halophyte plants.
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Abstract
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Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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FR1463385A FR3031040B1 (fr) | 2014-12-29 | 2014-12-29 | Procede d'obtention d'un extrait proteique concentre en superoxydase dismutase (sod) |
PCT/EP2015/081303 WO2016107850A1 (fr) | 2014-12-29 | 2015-12-28 | Procede d'obtention d'un extrait proteique concentre en superoxyde dismutase (sod) |
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EP3240565A1 true EP3240565A1 (fr) | 2017-11-08 |
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EP15817894.7A Withdrawn EP3240565A1 (fr) | 2014-12-29 | 2015-12-28 | Procede d'obtention d'un extrait proteique concentre en superoxyde dismutase (sod) |
Country Status (4)
Country | Link |
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US (1) | US20180051349A1 (fr) |
EP (1) | EP3240565A1 (fr) |
FR (1) | FR3031040B1 (fr) |
WO (1) | WO2016107850A1 (fr) |
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FR3052978B1 (fr) | 2016-06-23 | 2020-01-31 | Biotechmarine | Procede d'obtention d'un extrait d'armeria maritima enrichi en diglycosides et son utilisation en cosmetique |
FR3102365B1 (fr) * | 2019-10-24 | 2021-11-12 | Oreal | Utilisation d’un extrait deLimonium vulgarepour hydrater la peau et améliorer la fonction barrière. |
CN114507751B (zh) * | 2022-03-16 | 2023-07-21 | 新疆农业大学 | 一种与小麦籽粒超氧化物歧化酶活性相关的分子标记及其应用 |
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LU85910A1 (fr) * | 1985-05-22 | 1986-12-05 | Oleofina Sa | Procede pour eliminer l'oxygene dans les aliments et les boissons,et composition enzymatique utilisee a cet effet |
FR2729296B1 (fr) | 1995-01-12 | 1997-03-28 | Europlanaire | Compositions pharmaceutiques comprenant une superoxyde dismutase |
FR2747044B1 (fr) * | 1996-04-03 | 1998-06-26 | Coletica | 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 |
CN102289862B (zh) | 2011-08-17 | 2013-06-26 | 广州广电运通金融电子股份有限公司 | 闸门机构 |
EP2722075B1 (fr) * | 2012-10-22 | 2018-02-28 | BASF Beauty Care Solutions France S.A.S. | Prévention de conditions de peau sèche et apaisement pour peaux sensibles |
-
2014
- 2014-12-29 FR FR1463385A patent/FR3031040B1/fr active Active
-
2015
- 2015-12-28 US US15/540,723 patent/US20180051349A1/en not_active Abandoned
- 2015-12-28 EP EP15817894.7A patent/EP3240565A1/fr not_active Withdrawn
- 2015-12-28 WO PCT/EP2015/081303 patent/WO2016107850A1/fr active Application Filing
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Also Published As
Publication number | Publication date |
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FR3031040A1 (fr) | 2016-07-01 |
US20180051349A1 (en) | 2018-02-22 |
FR3031040B1 (fr) | 2018-09-07 |
WO2016107850A1 (fr) | 2016-07-07 |
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