EP1722803A1 - Extrapallial fluid of molluscs, method of obtaining same, formulation and use thereof - Google Patents

Extrapallial fluid of molluscs, method of obtaining same, formulation and use thereof

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Publication number
EP1722803A1
EP1722803A1 EP05717633A EP05717633A EP1722803A1 EP 1722803 A1 EP1722803 A1 EP 1722803A1 EP 05717633 A EP05717633 A EP 05717633A EP 05717633 A EP05717633 A EP 05717633A EP 1722803 A1 EP1722803 A1 EP 1722803A1
Authority
EP
European Patent Office
Prior art keywords
fluid
liquid
recovered
shell
mollusk
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Withdrawn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
EP05717633A
Other languages
German (de)
French (fr)
Inventor
Evelyne Lopez
Xavier Bourrat
Marthe Rousseau
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.)
D'innovation Et De Recherche Appliquee Sa (siera Sa) Ste
Original Assignee
D'innovation Et De Recherche Appliquee Sa (siera Sa) Ste
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 D'innovation Et De Recherche Appliquee Sa (siera Sa) Ste filed Critical D'innovation Et De Recherche Appliquee Sa (siera Sa) Ste
Publication of EP1722803A1 publication Critical patent/EP1722803A1/en
Withdrawn legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/56—Materials from animals other than mammals
    • A61K35/618—Molluscs, e.g. fresh-water molluscs, oysters, clams, squids, octopus, cuttlefish, snails or slugs

Definitions

  • the present invention relates to a process for obtaining a liquid comprising extracting from a mollusk the liquid located between the shell and the body of the mollusk, the liquid capable of being obtained by this process, its use in the pharmaceutical field, and in particular to improve skin healing, to regenerate tissues, or to promote osteogenesis or bone mineralization. It also relates to compositions, in particular pharmaceuticals, medical devices or food supplements containing it.
  • mother-of-pearl has made it possible to offer effective solutions in specific cases.
  • implants made from mother-of-pearl have been proposed for endosseous repairs.
  • Powdered mother-of-pearl has also been suggested as a filling material where it then shows biodissolution properties; there is a gradual replacement with newly formed compact and spongy bone depending on the nature of the recipient bone.
  • the use of mother-of-pearl as a biomaterial is unfortunately not always easy or possible in many cases, for example during large local repairs of degenerated tissue or loss of substances.
  • the applicant has just developed a technique for obtaining the extrapalleal fluid located between the shell and the body of a mollusk by extraction of said fluid (or liquid).
  • the fluid thus obtained can retain its self-organizing properties and that the latter can be used, even in a context external to the animal.
  • this fluid is of great industrial interest because of its physical nature, since it can be in many forms, such as for example in the form of a liquid, a film, a coating, fibers or in the form of a porous material. Thus, it can easily be handled and used in different forms, while retaining its biological properties.
  • the present invention therefore relates to a process for obtaining an extrapalleal fluid comprising recovering the extrapalleal fluid located between the interior of the shell and the body of a mollusk, in particular the mantle of said mollusk.
  • the fluid obtained is thus pure and taken in its entirety, in particular unmodified and unprocessed, after recovery, by a process modifying its intrinsic qualities. It can then be formulated, to be used in particular in a pharmaceutical composition, a medical device or food supplements.
  • the pharmaceutical composition according to the invention is in particular intended to improve the healing phenomena, to regenerate tissues, biological or bone, to promote osteogenesis or bone mineralization.
  • the method for preparing a liquid therefore comprises a step of recovering the extrapalleal fluid located between the shell and the body of a mollusk, in particular the mantle of said mollusk, said fluid having been extracted from 'a mollusk.
  • the fluid thus recovered corresponds to natural secretions of the mollusk which are found on the external surface of the mantle of the molluscs on the side of the shell.
  • this liquid even once extracted and therefore outside of its natural context, could be used for its regenerative and mineralizing properties, and thus promote the production of the shell (organomineralized material) of a mollusc, and more broadly substitute for, or regenerate, secretions from any other tissue known to form an extracellular matrix.
  • the liquid thus has the advantage of regenerating the tissues, biological or bone, in particular by stimulating the activity of the cells which form said tissues or by providing the basic materials useful for said regeneration, and by promoting osteogenesis or bone mineralization. .
  • tissue regeneration is meant the reconstruction and renewal of any tissue that has suffered damage or alteration (or injured tissue).
  • mineralization we mean the action (capacity) which consists in providing mineral elements or which favors the fixing of mineral elements on organic matter.
  • mineral species crystals, atoms, ions, molecules containing “mineral” elements and / or trace elements, as opposed to organic elements.
  • the regeneration of biological or bone tissue generally consists of reconstructing or renewing all or part of the altered tissue, in particular thanks to molecules of specific structure, to “signal” molecules and to organic and / or mineral nourishing elements present in the liquid obtained according to the invention. In addition, no intolerance was detected during the application of the liquid thus extracted and / or subsequently modified on or in a mammal.
  • the step of removing or extracting the liquid is generally carried out as follows.
  • the mollusc from which the liquid is extracted is generally a mono- or bi-valve mollusk, pearly or not, advantageously pearly.
  • the molluscs are in particular oysters and more specifically oysters of the genus Pinctada and more particularly of the genus Pinctada species margaritifera or maxima.
  • Oysters can also be chosen from those of the genus Ostrea, in particular of the species edulis, or of the genus Crassostrea, in particular of the species gigas. They may also be cephalopods, such as the nautilus, or gastropods, such as the periwinkle.
  • the mollusc used can come from aquaculture or land farming.
  • the mollusc can be sacrificed before, during or after the process according to the invention. It can be used dead or alive.
  • the mollusk for example can be sacrificed before the process according to the invention, taking care to keep at least partially (preferably completely) the liquid to be extracted in the shell.
  • the extraction process or step is a non-invasive method.
  • the liquid located between the interior of the shell and the body of the mollusk, in particular the mantle of said mollusk corresponds to natural secretions of the mollusk which are found on the external surface of the mantle of the molluscs on the side of the shell. and therefore which are outside the body of the mollusk proper.
  • the shell of the mollusc is preferably peeled off in whole or in part.
  • the mollusk is bi-valve, at least one of the two valves is raised so as to be able to access the body of the mollusk, prior to extraction.
  • the detachment (or separation) of the liquid from the shell can be carried out manually, generally using a suitable tool (such as: scraper or flexible spatula).
  • a suitable tool such as: scraper or flexible spatula
  • the detachment of the liquid from the shell is carried out chemically.
  • an enzymatic reaction advantageously non-denaturing, and generally at room temperature, can also be implemented so as to separate, more finely, the liquid from the shell.
  • this enzymatic reaction is carried out in the presence of at least one protease, such as for example trypsin.
  • the reaction can then be stopped by an anti-protease or by simple dilution.
  • the liquid is then withdrawn from the surface of the shell using an appropriate instrument, in particular using a syringe (in particular provided with a needle).
  • the liquid withdrawn is advantageously sterilized or decontaminated by any method known per se, in particular by filtration, for example on a Millipore® filter (example: 0.20 ⁇ m). It can also be dehydrated, in particular by lyophilization. Liquid can also be frozen, preferably just after recovery or after filtration.
  • the sampling can be carried out on living molluscs, for example in the case of large molluscs, such as pearly bivalves of the Pinctada family.
  • This variant preferably consists in keeping the shell of the living bivalve open using an instrument, such as a wedge and / or forceps forceps (or retractor).
  • the instrument is preferably chosen so as not to injure the animal, in particular it is chosen so as not to irreversibly damage its metabolism.
  • the mantle is preferably lifted in whole or in part to separate it from at least one of the shells.
  • the liquid is then taken from the surface of the shell using an appropriate instrument, such as in particular a scraper or flexible spatula.
  • a scraper plastic
  • the liquid is harvested using a syringe generally equipped with a needle, preferably with a diameter between 1 mm and 5 mm.
  • any other material suitable for harvesting the liquid can be considered.
  • the process for obtaining the fluid according to the invention may also comprise subsequent non-denaturing steps of the fluid thus recovered, such as steps of filtration, decontamination, sterilization, freezing / thawing, changes in viscosity, and / or various layouts.
  • the liquid obtained according to the method described above can be used such that, in its pure state (taken in its entirety), in particular it is unmodified and unprocessed, after recovery, by a process modifying its intrinsic qualities (denaturing stages), in particular by a demineralization process.
  • the liquid obtained can be formulated subsequently according to its subsequent use.
  • the subsequent use of the liquid may require a suitable method of preparation.
  • Experiments carried out with the liquid thus recovered show that it is very flexible to handle and that it adapts very easily to all kinds of constraints linked to its implementation. It can thus undergo several thawing and re-freezing without affecting its biological properties. It is the same for lyophilization.
  • a person skilled in the art will take care not to modify the intrinsic qualities of the liquid thus extracted and will therefore avoid subjecting this liquid to any operation detrimental to its interest, such as a demineralization operation.
  • the fluid may for example be in the form of a film or a coating.
  • Any method known per se for forming a liquid or a film can be used. It can for example be spread on a smooth or porous surface or any other support and form a film (film-forming character of the fluid obtained).
  • the film or coating thus formed retains the regenerating and healing character of the liquid. It can thus constitute an organic biological film on the surface, with a mineralizing power.
  • the thickness of the film can vary to a large extent depending on its end use.
  • a thick film can for example be obtained with the triple property: (i) sealing, (ii) self-supporting structure, (iii) intrinsic mineral contribution, while having the regenerating and healing character.
  • This material formed by an intimate association between organic matter and mineral matter, forms a film on any surface and crystallizes polymorphically depending on the condition and texture of the surface.
  • the film thus obtained can be used on its support or be removed from the support for later use.
  • the liquid can also be used later in the form of fibers. Any method known per se for forming a liquid into fibers can be used.
  • This liquid has the advantage of being spinnable over a wide range of viscosities. In particular, a very simple stretching technique, in particular using tweezers, makes it possible to stretch fibers from the liquid. It is possible in this way to use it in situ (locally) or in another way, as a porous, invasive filling material, to fill as needed a given volume.
  • the invasive nature of this material allows in particular biological materials, such as cells, to penetrate there and exert their effects in association with the properties of the shaped liquid.
  • the same material can be used to bridge two distant tissue banks.
  • the same principle can be applied to very large volumes, in particular by coating fibers of another chemical nature (for example carbon fibers).
  • the fibers thus coated have a coating on their surface comprising the constituent elements of the liquid according to the invention and can be used to provide, for example, a healing interface.
  • the fluid can thus come in different forms, including liquid, films, coatings and fibers.
  • it can also be in the form of a porous material, in particular by using, during its shaping, a porous support.
  • the porous material may be a film, a coating or be in any other form, such as a spherical shape.
  • the fluid according to the invention generally comprises a mixture of the constituent elements of living matter, in particular a mixture of proteins, proteoglycans, polysaccharides, lipids and mineral salts, this is more particularly the case for a mother-of-pearl mollusc, advantageously an oyster of the genus Pinctada (generally taken in its natural biotope).
  • a mother-of-pearl mollusc advantageously an oyster of the genus Pinctada (generally taken in its natural biotope).
  • the nature and quantity of these elements can vary to a large extent, particularly in function of the mollusc, its biotope, the season of the sample and the conditions of the sample.
  • composition of this liquid preferably comprises at least the following compounds: proteins in an average amount of 0.5 mg / mL, proteoglycans in an average amount of 3 mg / mL, polysaccharides in an average amount of 1.5 mg / mL , lipids in an average amount of 1 mg / mL and mineral salts in an average amount of 7.5 mg / mL.
  • a quantitative analysis of the trace elements present in a specific liquid (identical to that identified above) according to the invention was carried out, in particular by quantitatively measuring the trace elements present by analysis by neutron activation (ANA) and spectrometry of mass coupled to a plasma torch (ICP-MS).
  • ANA neutron activation
  • ICP-MS spectrometry of mass coupled to a plasma torch
  • composition of this liquid preferably comprises the trace elements chosen from: S, Mg, La, Zn, Br, Ce, Fe, Mn, Cl, Cu, K, Sr, Na and Ca
  • trace elements chosen from: S, Mg, La, Zn, Br, Ce, Fe, Mn, Cl, Cu, K, Sr, Na and Ca
  • the amounts of these trace elements are respectively as follows: 0.02; 2; 0.28; 0.4; 1.78; 5.5; 13.6; 50; 296; 143; 582; 1000; 5420 (expressed in ⁇ g / g) and 38.8 (expressed in ⁇ g / 100g).
  • a quantitative analysis of the amino acids present in free form or bound in a fluid according to the invention was carried out, in particular by quantitatively measuring the amino acids present by the amino acid analyzer Beckmann 6300 using the conventional method with ninhydrin. The result of this analysis is given in FIG. 1 (% by weight of amino acids relative to the total weight of amino acids).
  • the present invention also relates to the fluid capable of being obtained by the process as described above.
  • the liquid (or fluid) can be not only the liquid obtained directly after the recovery stage described above, but also that obtained after subsequent non-denaturing stages, such as stages of filtration, decontamination, sterilization, freezing / thawing, changes in viscosity, and / or various forms, such as in particular those identified above.
  • the fluid according to the invention is used as a medicament.
  • the present invention also relates to a composition, in particular pharmaceutical, characterized in that it comprises the fluid as defined above, in particular in combination with a pharmaceutically acceptable excipient.
  • the liquid used as a medicament or in the composition detailed below may be that obtained directly after the recovery stage described above, but also that obtained after subsequent stages of filtration, decontamination, sterilization, freezing / thawing, changes in viscosity, and / or various forms, such as in particular those identified above.
  • the administration of the pharmaceutical composition or of the medicament according to the invention can be carried out by topical, enteral or parenteral route.
  • the pharmaceutical composition is packaged in a form suitable for topical application, that is to say applied in situ to the place where the tissue is injured.
  • the pharmaceutical composition can be in the form of tablets, capsules, dragees, syrups, suspensions, solutions, powders, granules, emulsions, microspheres or nanospheres or vesicles lipidic or polymeric allowing controlled release.
  • the composition can be in the form of solutions or suspensions for infusion or for injection.
  • the pharmaceutical composition is intended for local use on or in the injured tissue.
  • the pharmaceutically acceptable carrier is in particular an excipient suitable for topical application.
  • the pharmaceutical composition according to the invention is more particularly intended for the treatment of the tissues mentioned above, in particular for the treatment of the skin and bones of a mammal, in particular human. It can be in the form of ointments, creams, milks, ointments, powders, soaked pads, solutions, gels, sprays, lotions or suspensions. It can also be in the form of microspheres or nanospheres or lipid or polymeric vesicles or of polymeric patches and hydrogels allowing controlled release.
  • This topical composition can be in anhydrous form, in aqueous form or in the form of an emulsion (water / oil, oil / water emulsion or multiple emulsion).
  • the liquid according to the invention is used, preferably topically, at a concentration generally between 0.02% and 20% by weight, preferably between 0.25 and 10% by weight, and advantageously between 0.5 and 5%, relative to the total weight of the composition.
  • the therapeutic composition according to the invention is characterized in that it contains, in association with inert excipients, a therapeutically effective amount of the liquid according to the invention, in particular for improving skin healing, for regenerating the tissues, or to promote osteogenesis or bone mineralization.
  • Tissues include soft tissue or skeletal, including in particular the skeleton, bones, cartilage, teeth, or any other tissue, in particular the skin and its appendages.
  • the composition according to the invention makes it possible to treat in particular bone, cartilaginous or dental disorders.
  • the invention also resides in a method for improving skin healing, for regenerating tissues, in particular those identified above, or for promoting osteogenesis or bone mineralization, comprising administration to subjects in need of such treatment a therapeutically effective amount of the liquid as defined above.
  • treatment designates preventive, curative, palliative treatment, as well as the management of patients (reduction of pain, improvement of the quality of life, slowing down of the progression of the disorder, trauma or illness), etc.
  • the method or treatment can also be carried out in combination with other ingredients or treatments, such as in particular other active compounds to improve skin healing, to regenerate tissues, or to promote osteogenesis or bone mineralization.
  • compositions or medicaments according to the invention can also comprise at least one other therapeutically active ingredient, for simultaneous, separate or spread over time use.
  • compositions or medicaments according to the invention advantageously comprise one or more inert excipients or vehicles, that is to say pharmaceutically inactive and non-toxic. Mention may be made, for example, of saline, physiological, isotonic, buffered solutions, etc., compatible with pharmaceutical use and known to those skilled in the art.
  • the essays may contain one or more agents or vehicles chosen from dispersants, solubilizers, stabilizers, preservatives, etc.
  • Agents or vehicles usable in formulations are in particular methylcellulose, Phydroxymethylcellulose, carboxymethylcellulose, cylclodextrins, polysorbate 80, mannitol, gelatin, lactose, vegetable or animal oils , acacia, etc.
  • the compositions can be formulated in the form of an injectable suspension, gels, oils, tablets, suppositories, powders, capsules, capsules, etc., optionally by means of dosage forms or devices ensuring sustained and / or delayed release.
  • at least one agent such as cellulose, carbonates or starches is used.
  • compositions as described above can of course also contain pharmacodynamically active additives or a combination of these additives, and in particular: wetting agents, emollients, hydrating agents, such as glycerol, PEG 400 or else urea .
  • compositions may also contain flavor enhancers, preservatives such as parahydroxybenzoic acid esters, stabilizers, moisture regulators, pH regulators, osmotic pressure modifiers , emulsifying agents, antioxidants, such as ⁇ -tocopherol, butylhydroxyanisole or butylhydroxytoluene.
  • flavor enhancers such as parahydroxybenzoic acid esters, stabilizers, moisture regulators, pH regulators, osmotic pressure modifiers , emulsifying agents, antioxidants, such as ⁇ -tocopherol, butylhydroxyanisole or butylhydroxytoluene.
  • the administration can be carried out by any method known to those skilled in the art, in particular by the oral, topical or injection route, typically by the intraperitoneal, intra-cerebral, intra-thecal, intravenous, intra-arterial or intramuscular route. Topical or oral administration is preferred. For long-term, non-topical treatment, the preferred route of administration will be sublingual, oral or transcutaneous.
  • the compounds are generally packaged in the form of liquid suspensions, which can be injected using syringes or infusions, for example. It is understood that the flow rate and / or the dose injected, or in general the dose to be administered, can be adapted by a person skilled in the art according to the subject to be treated, the disorder, the mode of administration, etc. It is understood that repeated administrations can be carried out, optionally in combination with other active ingredients or any pharmaceutically acceptable vehicle (buffers, saline, isotonic solutions, in the presence of stabilizing agents, etc.).
  • the invention relates to a device, more specifically suitable for subcutaneous or percutaneous injection, comprising the liquid as defined above and a physiologically acceptable excipient or support.
  • This device can in particular be in the form of syringes or infusions.
  • the medical device of the present invention may be intended to be implanted in or on the body of a mammal, the liquid as defined above may be located inside a membrane suitable for this use .
  • the invention relates to a food supplement comprising the liquid as defined above.
  • This food supplement is in particular intended to improve cutaneous healing, to regenerate tissues, or to favor osteogenesis or mineralization of bones or teeth.
  • the invention can be used in mammals, in particular in humans. Other aspects and advantages of the present invention will appear on reading the examples which follow, which should be considered as illustrative and not limiting.
  • Example 1 Recovery of the liquid from a sacrificed animal The mollusk is sacrificed so that the liquid remains in the shell. We take off the coat carefully; an enzymatic reaction is then used to completely detach the liquid from the shell. The reaction is then stopped by an anti-protease. The liquid is then taken from the surface of the shell. As soon as collected, the liquid can be frozen. It can be sterilized by filtration on a 0.20 ⁇ m millipore and can be freeze-dried while retaining its properties.
  • An oyster of the species of Pinctada margaritifera is half-opened thanks to a retractor according to the practices well known to those skilled in the art in pearl farming.
  • the excess water still present is discharged.
  • the animal is arranged horizontally.
  • the mantle is delicately separated from the lower shell using forceps, then the liquid is withdrawn using a syringe, for example.
  • the liquid is: - filtered - lyophilized - a few weeks later, the liquid is reformed with sterile water - it is spread in the form of a thick film.
  • the oyster is fixed and the coat is peeled off. In this way, it is possible to collect the liquid from the first valve. The animal is then turned over to operate on the other side. In this case, an amount of 50 to 100 ⁇ l per valve is obtained.
  • the liquid collected from a population of around twenty animals is subjected to conditioning similar to that of Example 2.
  • the liquid is: - filtered - lyophilized - a few weeks later, the liquid is reformed with sterile water - it is spread in the form of a thick film
  • the liquid is reconstituted even after lyophilization, that it forms a film on a polypropylene support.
  • This film solidifies by self-organization and self-healing.

Abstract

The invention relates to a fluid-preparation method comprising the recovery of the fluid that is located between the shell and the body of a mollusc. The invention also relates to the liquid obtained using said method and to the use thereof in the pharmaceutical field, for example, in order to improve cutaneous healing, to regenerate tissues or to promote osteogenesis or bone mineralisation. The invention further relates to compositions, particularly pharmaceutical compositions, medical devices or food supplements containing same.

Description

LIQUIDES EXTRAPALLEAUX DE MOLLUSQUES. OBTENTION. FORMULATION ET UTILISATION EXTRAPALLEL LIQUIDS OF MOLLUSCS. OBTAINING. FORMULATION AND USE
La présente invention concerne un procédé d'obtention d'un liquide comprenant l'extraction à partir d'un mollusque du liquide se situant entre la coquille et le corps du mollusque, le liquide susceptible d'être obtenu par ce procédé, son utilisation dans le domaine pharmaceutique, et notamment pour améliorer la cicatrisation cutanée, pour régénérer les tissus, ou pour favoriser l'ostéogenèse ou la minéralisation osseuse. Elle concerne également des compositions, en particulier pharmaceutiques, des dispositifs médicaux ou des compléments alimentaires le contenant.The present invention relates to a process for obtaining a liquid comprising extracting from a mollusk the liquid located between the shell and the body of the mollusk, the liquid capable of being obtained by this process, its use in the pharmaceutical field, and in particular to improve skin healing, to regenerate tissues, or to promote osteogenesis or bone mineralization. It also relates to compositions, in particular pharmaceuticals, medical devices or food supplements containing it.
Dans de nombreux cas de dégénérescence des tissus minéralisés, la réparation tissulaire reste un problème complexe non encore résolu, notamment à cause de la difficulté de mettre en œuvre des matériaux de substitution induisant une régénération. Parmi ces matériaux de substitution induisant une régénération, la nacre a déjà montré sa biocompatibilité, sa capacité ostéogénique et ostéoinductrice vis-à-vis des ostéoblastes humains [Lopez et al., Tissue & Cell, 1992 : 667-679 ; Lamghari et al., J. Bone Miner. Res. 2001 ; 16 : 2232-2237].In many cases of degeneration of mineralized tissues, tissue repair remains a complex problem that has not yet been resolved, in particular because of the difficulty of using substitute materials inducing regeneration. Among these substitution materials inducing regeneration, mother-of-pearl has already shown its biocompatibility, its osteogenic and osteoinductive capacity vis-à-vis human osteoblasts [Lopez et al., Tissue & Cell, 1992: 667-679; Lamghari et al., J. Bone Miner. Res. 2001; 16: 2232-2237].
Dans ce cadre, la nacre a permis de proposer des solutions efficaces dans des cas particuliers. Ainsi, des implants fabriqués à partir de nacre ont été proposés pour des réparations endo-osseuses. La nacre réduite en poudre a également été suggérée comme matériau de comblement où elle montre alors des propriétés de biodissolution ; on observe un remplacement progressif par de l'os néoformé compact et spongieux suivant la nature de l'os receveur. L'utilisation de la nacre comme biomatériau n'est malheureusement pas toujours aisée ou possible dans de nombreux cas, par exemple lors de larges réparations locales de tissus dégénérés ou de pertes de substances.In this context, mother-of-pearl has made it possible to offer effective solutions in specific cases. Thus, implants made from mother-of-pearl have been proposed for endosseous repairs. Powdered mother-of-pearl has also been suggested as a filling material where it then shows biodissolution properties; there is a gradual replacement with newly formed compact and spongy bone depending on the nature of the recipient bone. The use of mother-of-pearl as a biomaterial is unfortunately not always easy or possible in many cases, for example during large local repairs of degenerated tissue or loss of substances.
II n'existe donc pas à ce jour de biomatériau aisément formulable et permettant de régénérer les tissus biologiques ou osseux de manière totalement satisfaisante.To date, there is therefore no easily formulated biomaterial which makes it possible to regenerate biological or bone tissues in a completely satisfactory manner.
A ce titre, la demanderesse vient de mettre au point une technique d'obtention du fluide extrapalleal se situant entre la coquille et le corps d'un mollusque par extraction dudit fluide (ou liquide). La demanderesse vient de mettre en évidence que le fluide ainsi obtenu peut conserver ses propriétés auto-organisantes et que ces dernières peuvent être utilisées, même dans un contexte externe à l'animal. En outre, ce fluide présente un grand intérêt industriel du fait de sa nature physique, puisqu'il peut se présenter sous de nombreuses formes, comme par exemple sous la forme d'un liquide, d'un film, d'un revêtement, de fibres ou encore sous la forme d'un matériau poreux. Ainsi, il peut facilement être manipulé et utilisé sous différentes formes, tout en gardant ses propriétés biologiques.As such, the applicant has just developed a technique for obtaining the extrapalleal fluid located between the shell and the body of a mollusk by extraction of said fluid (or liquid). The Applicant has just demonstrated that the fluid thus obtained can retain its self-organizing properties and that the latter can be used, even in a context external to the animal. In addition, this fluid is of great industrial interest because of its physical nature, since it can be in many forms, such as for example in the form of a liquid, a film, a coating, fibers or in the form of a porous material. Thus, it can easily be handled and used in different forms, while retaining its biological properties.
La présente invention vise donc un procédé d'obtention d'un fluide extrapalleal comprenant la récupération du fluide extrapalleal se situant entre l'intérieur de la coquille et le corps d'un mollusque, en particulier le manteau dudit mollusque.The present invention therefore relates to a process for obtaining an extrapalleal fluid comprising recovering the extrapalleal fluid located between the interior of the shell and the body of a mollusk, in particular the mantle of said mollusk.
Elle a également pour objet le fluide susceptible d'être obtenu par ce procédé et son utilisation dans divers domaines. Le fluide obtenu est ainsi pur et pris dans son intégralité, en particulier non modifié et non transformé, ultérieurement à la récupération, par un procédé modifiant ses qualités intrinsèques. Il peut être par la suite formulé, pour être en particulier utilisé dans une composition pharmaceutique, un dispositif médical ou des compléments alimentaires. La composition pharmaceutique selon l'invention est en particulier destinée à améliorer les phénomènes de cicatrisation, à régénérer les tissus, biologiques ou osseux, favoriser l'ostéogenèse ou la minéralisation osseuse.It also relates to the fluid capable of being obtained by this process and its use in various fields. The fluid obtained is thus pure and taken in its entirety, in particular unmodified and unprocessed, after recovery, by a process modifying its intrinsic qualities. It can then be formulated, to be used in particular in a pharmaceutical composition, a medical device or food supplements. The pharmaceutical composition according to the invention is in particular intended to improve the healing phenomena, to regenerate tissues, biological or bone, to promote osteogenesis or bone mineralization.
Chez l'animal, ce liquide participe in situ à la minéralisation de l'exosquelette. Certains voient une grande unicité des processus de minéralisation développés par la nature, depuis les exosquelettes des mollusques jusqu'à l'endosquelette des mammifères supérieurs. Le champ d'application de ces bioproduits est donc potentiellement très étendu, dans les domaines squelettiques, os et cartilage, dents, ou tout autre tissu, en particulier la peau et ses phanères.In animals, this liquid participates in situ in the mineralization of the exoskeleton. Some see a great uniqueness of the mineralization processes developed by nature, from the exoskeletons of molluscs to the endoskeletons of higher mammals. The field of application of these bioproducts is therefore potentially very wide, in the skeletal fields, bones and cartilage, teeth, or any other tissue, in particular the skin and its integuments.
Selon l'invention, le procédé de préparation d'un liquide comprend donc une étape de récupération du fluide extrapalleal se situant entre la coquille et le corps d'un mollusque, en particulier le manteau dudit mollusque, ledit fluide ayant été extrait à partir d'un mollusque.According to the invention, the method for preparing a liquid therefore comprises a step of recovering the extrapalleal fluid located between the shell and the body of a mollusk, in particular the mantle of said mollusk, said fluid having been extracted from 'a mollusk.
Le fluide ainsi récupéré correspond à des sécrétions naturelles du mollusque qui se trouvent à la surface externe du manteau des mollusques du coté de la coquille. De manière surprenante, la demanderesse a constaté que ce liquide, même une fois extrait et donc en dehors de son contexte naturel, pouvait être utilisé pour ses propriétés régénératrices et minéralisantes, et promouvoir ainsi la fabrication de la coquille (matière organominéralisée) d'un mollusque, et de façon plus large se substituer à, ou régénérer, des sécrétions de tout autre tissu connu pour former une matrice extracellulaire. Le liquide présente ainsi l'avantage de régénérer les tissus, biologiques ou osseux, notamment en stimulant l'activité des cellules qui forment lesdits tissus ou en apportant les matériaux de base utiles à ladite régénération, et de favoriser l'ostéogenèse ou la minéralisation osseuse.The fluid thus recovered corresponds to natural secretions of the mollusk which are found on the external surface of the mantle of the molluscs on the side of the shell. Surprisingly, the Applicant has found that this liquid, even once extracted and therefore outside of its natural context, could be used for its regenerative and mineralizing properties, and thus promote the production of the shell (organomineralized material) of a mollusc, and more broadly substitute for, or regenerate, secretions from any other tissue known to form an extracellular matrix. The liquid thus has the advantage of regenerating the tissues, biological or bone, in particular by stimulating the activity of the cells which form said tissues or by providing the basic materials useful for said regeneration, and by promoting osteogenesis or bone mineralization. .
Par régénération de tissus, on entend la reconstruction et le renouvellement de tout tissu ayant subi un dommage ou une altération (ou tissu lésé). Par minéralisation (minéralisateur ou minéralisant), on entend l'action (capacité) qui consiste à apporter des éléments minéraux ou qui favorise la fixation des éléments minéraux sur de la matière organique. Par espèces minérales, on entend les cristaux, les atomes, les ions, les molécules contenant des éléments « minéraux » et/ou oligo- éléments, par opposition aux éléments organiques. Ainsi, la régénération d'un tissu biologique ou osseux consiste généralement à reconstruire ou renouveler en tout ou partie le tissu altéré, notamment grâce aux molécules de structure spécifique, aux molécules « signal » et aux éléments nourriciers organiques et/ou minéraux présents dans le liquide obtenu selon l'invention. En outre, aucune intolérance n'a été détectée lors de l'application du liquide ainsi extrait et/ou modifié ultérieurement sur ou dans un mammifère.By tissue regeneration is meant the reconstruction and renewal of any tissue that has suffered damage or alteration (or injured tissue). By mineralization (mineralizer or mineralizer), we mean the action (capacity) which consists in providing mineral elements or which favors the fixing of mineral elements on organic matter. By mineral species is meant crystals, atoms, ions, molecules containing “mineral” elements and / or trace elements, as opposed to organic elements. Thus, the regeneration of biological or bone tissue generally consists of reconstructing or renewing all or part of the altered tissue, in particular thanks to molecules of specific structure, to “signal” molecules and to organic and / or mineral nourishing elements present in the liquid obtained according to the invention. In addition, no intolerance was detected during the application of the liquid thus extracted and / or subsequently modified on or in a mammal.
L'étape de prélèvement ou d'extraction du liquide est généralement réalisée de la manière suivante. Le mollusque à partir duquel est extrait le liquide est généralement un mollusque mono- ou bi-valve, nacrier ou non, avantageusement nacrier. Les mollusques sont en particulier des huîtres et plus spécifiquement des huîtres du genre Pinctada et plus particulièrement du genre Pinctada espèce margaritifera ou maxima. Les huitres peuvent être aussi choisies parmi celles du genre Ostrea, en particulier de l'espèce edulis, ou encore du genre Crassostrea, en particulier de l'espèce gigas. Il peut s'agir également de céphalopodes, comme par exemple le nautile, ou encore de gastéropodes, comme par exemple le bigorneau.The step of removing or extracting the liquid is generally carried out as follows. The mollusc from which the liquid is extracted is generally a mono- or bi-valve mollusk, pearly or not, advantageously pearly. The molluscs are in particular oysters and more specifically oysters of the genus Pinctada and more particularly of the genus Pinctada species margaritifera or maxima. Oysters can also be chosen from those of the genus Ostrea, in particular of the species edulis, or of the genus Crassostrea, in particular of the species gigas. They may also be cephalopods, such as the nautilus, or gastropods, such as the periwinkle.
Le mollusque utilisé peut provenir d'un élevage aquacole ou terrestre. Le mollusque peut être sacrifié avant, pendant ou après le procédé selon l'invention. Il peut être utilisé mort ou vivant. Le mollusque par exemple peut être sacrifié avant le procédé selon l'invention, en prenant garde de conserver au moins partiellement (de préférence totalement) le liquide à extraire dans la coquille.The mollusc used can come from aquaculture or land farming. The mollusc can be sacrificed before, during or after the process according to the invention. It can be used dead or alive. The mollusk for example can be sacrificed before the process according to the invention, taking care to keep at least partially (preferably completely) the liquid to be extracted in the shell.
Le prélèvement du fluide extrapalleal peut être réalisé sur des lots de mollusques exclusivement destinés à cet usage, sans nuire pour autant à leur intégralité. Les mollusques peuvent être éventuellement utilisés par la suite, ceux-ci restant saints et vivants.Sampling of extrapalleal fluid can be carried out on batches of molluscs exclusively intended for this use, without harming their entirety. The molluscs can eventually be used, these remaining holy and alive.
De manière plus spécifique, le procédé ou l'étape d'extraction est une méthode non invasive. En effet, le liquide se situant entre l'intérieur de la coquille et le corps du mollusque, en particulier le manteau dudit mollusque, correspond à des sécrétions naturelles du mollusque qui se trouvent à la surface externe du manteau des mollusques du coté de la coquille et donc qui se trouvent à l'extérieur du corps du mollusque proprement dit.More specifically, the extraction process or step is a non-invasive method. In fact, the liquid located between the interior of the shell and the body of the mollusk, in particular the mantle of said mollusk, corresponds to natural secretions of the mollusk which are found on the external surface of the mantle of the molluscs on the side of the shell. and therefore which are outside the body of the mollusk proper.
Afin d'accéder au liquide à extraire, le manteau du mollusque est de préférence décollé en tout ou partie. Lorsque le mollusque est bi-valve, au moins une des deux valves est soulevée de manière à pouvoir accéder au corps du mollusque, préalablement à l'extraction.In order to access the liquid to be extracted, the shell of the mollusc is preferably peeled off in whole or in part. When the mollusk is bi-valve, at least one of the two valves is raised so as to be able to access the body of the mollusk, prior to extraction.
Le décollement (ou séparation) du liquide de la coquille peut être réalisé manuellement, généralement à l'aide d'un outil approprié (comme par exemple : racloir ou spatule souple). Selon une autre méthode, éventuellement en complément dudit décollement manuel, le décollement du liquide de la coquille est réalisé chimiquement. Ainsi, une réaction enzymatique, avantageusement non dénaturante, et généralement à température ambiante, peut aussi être mise en œuvre de façon à séparer, plus finement, le liquide de la coquille. De préférence, cette réaction enzymatique est réalisée en présence d'au moins une protéase, telle que par exemple la trypsine. La réaction peut ensuite être stoppée par une anti- protéase ou par simple dilution.The detachment (or separation) of the liquid from the shell can be carried out manually, generally using a suitable tool (such as: scraper or flexible spatula). According to another method, possibly in addition to said manual detachment, the detachment of the liquid from the shell is carried out chemically. Thus, an enzymatic reaction, advantageously non-denaturing, and generally at room temperature, can also be implemented so as to separate, more finely, the liquid from the shell. Preferably, this enzymatic reaction is carried out in the presence of at least one protease, such as for example trypsin. The reaction can then be stopped by an anti-protease or by simple dilution.
Le liquide est alors prélevé à la surface de la coquille à l'aide d'un instrument approprié, notamment à l'aide d'une seringue (en particulier munie d'une aiguille). Le liquide prélevé est avantageusement stérilisé ou décontaminé par toute méthode connue en soi, notamment par filtration, par exemple sur filtre Millipore® (exemple : 0,20μm). Il peut être aussi déshydraté, notamment par lyophilisation. Le liquide peut également être congelé, de préférence juste après la récupération ou après filtration.The liquid is then withdrawn from the surface of the shell using an appropriate instrument, in particular using a syringe (in particular provided with a needle). The liquid withdrawn is advantageously sterilized or decontaminated by any method known per se, in particular by filtration, for example on a Millipore® filter (example: 0.20 μm). It can also be dehydrated, in particular by lyophilization. Liquid can also be frozen, preferably just after recovery or after filtration.
Selon une variante du procédé, le prélèvement peut être effectué sur des mollusques vivants, par exemple lorsqu'il s'agit de gros mollusques, comme les bivalves nacriers de la famille des Pinctada.According to a variant of the process, the sampling can be carried out on living molluscs, for example in the case of large molluscs, such as pearly bivalves of the Pinctada family.
Cette variante consiste de préférence à maintenir la coquille du bivalve vivant ouverte à l'aide d'un instrument, tel qu'une cale et/ou une pince forceps (ou ecarteur). L'instrument est de préférence choisi de manière à ne pas blesser l'animal, en particulier il est choisi de manière à ne pas léser de façon irréversible son métabolisme. Le manteau est préférentiellement soulevé en tout ou partie pour le séparer d'au moins de l'une des coquilles. Le liquide est alors prélevé à la surface de la coquille à l'aide d'un instrument approprié, tel que notamment racloir ou spatule souple. Avantageusement, un racloir (en plastique) est utilisé pour récolter le liquide se trouvant entre le manteau et la coquille. Alternativement, la récolte du liquide s'opère à l'aide d'une seringue généralement munie d'une aiguille, de préférence de diamètre compris entre 1 mm et 5 mm. Bien entendu, tout autre matériel adapté à la récolte du liquide peut être envisagé.This variant preferably consists in keeping the shell of the living bivalve open using an instrument, such as a wedge and / or forceps forceps (or retractor). The instrument is preferably chosen so as not to injure the animal, in particular it is chosen so as not to irreversibly damage its metabolism. The mantle is preferably lifted in whole or in part to separate it from at least one of the shells. The liquid is then taken from the surface of the shell using an appropriate instrument, such as in particular a scraper or flexible spatula. Advantageously, a scraper (plastic) is used to collect the liquid located between the mantle and the shell. Alternatively, the liquid is harvested using a syringe generally equipped with a needle, preferably with a diameter between 1 mm and 5 mm. Of course, any other material suitable for harvesting the liquid can be considered.
Le procédé d'obtention du fluide selon l'invention peut comprendre en outre des étapes ultérieures non dénaturantes du fluide ainsi récupéré, telles que des étapes de filtration, de décontamination, de stérilisation, de congélation/décongélation, de modifications de viscosité, et/ou de mises en formes variées.The process for obtaining the fluid according to the invention may also comprise subsequent non-denaturing steps of the fluid thus recovered, such as steps of filtration, decontamination, sterilization, freezing / thawing, changes in viscosity, and / or various layouts.
Le liquide obtenu selon la méthode décrite ci-dessus peut être utilisé tel que, à l'état pur (pris dans son intégralité), en particulier il est non modifié et non transformé, ultérieurement à la récupération, par un procédé modifiant ses qualités intrinsèques (étapes dénaturantes), notamment par un procédé de déminéralisation. Le liquide obtenu peut être formulé par la suite en fonction de son usage ultérieur. Ainsi, l'utilisation ultérieure du liquide peut nécessiter un mode de préparation adapté. Les expériences conduites avec le liquide ainsi récupéré montrent qu'il est d'un maniement très souple et qu'il s'adapte très facilement à toutes sortes de contraintes liées à sa mise en oeuvre. Il peut ainsi subir plusieurs décongélations et re-congélations sans que ses propriétés biologiques en soient affectées. Il en est de même pour la lyophilisation. Bien entendu, l'homme du métier veillera à ne pas modifier les qualités intrinsèques du liquide ainsi extrait et évitera donc de faire subir à ce liquide toute opération nuisant à son intérêt, tel qu'une opération de déminéralisation.The liquid obtained according to the method described above can be used such that, in its pure state (taken in its entirety), in particular it is unmodified and unprocessed, after recovery, by a process modifying its intrinsic qualities (denaturing stages), in particular by a demineralization process. The liquid obtained can be formulated subsequently according to its subsequent use. Thus, the subsequent use of the liquid may require a suitable method of preparation. Experiments carried out with the liquid thus recovered show that it is very flexible to handle and that it adapts very easily to all kinds of constraints linked to its implementation. It can thus undergo several thawing and re-freezing without affecting its biological properties. It is the same for lyophilization. Of course, a person skilled in the art will take care not to modify the intrinsic qualities of the liquid thus extracted and will therefore avoid subjecting this liquid to any operation detrimental to its interest, such as a demineralization operation.
Il est également possible de varier la viscosité du liquide ainsi obtenu en modifiant sa concentration en eau. On peut y incorporer un gélifiant ou tout autre substance facilitant son utilisation ultérieure, pour le rendre plus ou moins liquide (visqueux).It is also possible to vary the viscosity of the liquid thus obtained by modifying its water concentration. One can incorporate a gelling agent or any other substance facilitating its subsequent use, to make it more or less liquid (viscous).
Le fluide peut par exemple être mis sous forme de film ou de revêtement. Toute méthode connue en soi pour mettre sous forme de film ou de revêtement un liquide peut être mise en oeuvre. Il peut par exemple être étalé sur une surface lisse ou poreuse ou tout autre support et former un film (caractère filmogène du fluide obtenu). Le film ou revêtement ainsi formé conserve le caractère régénérant et cicatrisant du liquide. Il peut constituer ainsi un film biologique organique en surface, avec un pouvoir minéralisant. L'épaisseur du film peut varier dans une large mesure en fonction de son usage final. Un film épais peut être par exemple obtenu avec la triple propriété : (i) étanchéité, (ii) structure auto-supportée, (iii) apport minéral intrinsèque, tout en présentant le caractère régénérant et cicatrisant. Ce matériau, constitué par une association intime entre matière organique et matière minérale, forme un film sur toute surface et cristallise de façon polymorphe suivant l'état et la texture de la surface. Le film ainsi obtenu peut être utilisé sur son support ou être retiré du support pour un usage ultérieur. Le liquide peut aussi être utilisé ultérieurement sous forme de fibres. Toute méthode connue en soi pour mettre sous forme de fibres un liquide peut être mise en oeuvre. Ce liquide présente en effet l'avantage d'être filable dans un domaine large de viscosités. En particulier, une technique très simple d'étirement, notamment à l'aide d'une pince brucelles, permet d'étirer des fibres à partir du liquide. Il est possible de cette manière de l'utiliser in situ (localement) ou d'une autre manière, comme matériau de comblement poreux, invasif, pour remplir selon le besoin un volume donné. Le caractère invasif de ce matériau permet notamment à des matériaux biologiques, tels que des cellules, d'y pénétrer et d'y exercer leurs effets en association avec les propriétés du liquide mis en forme. Le même matériau peut être utilisé pour ponter deux berges tissulaires éloignées. Le même principe peut s'appliquer sur de très gros volumes, notamment par enduction de fibres d'une autre nature chimique (par exemple des fibres de carbone). Les fibres ainsi enduites présentent un revêtement à leur surface comprenant les éléments constitutifs du liquide selon l'invention et peuvent être utilisées pour assurer par exemple une interface cicatrisante.The fluid may for example be in the form of a film or a coating. Any method known per se for forming a liquid or a film can be used. It can for example be spread on a smooth or porous surface or any other support and form a film (film-forming character of the fluid obtained). The film or coating thus formed retains the regenerating and healing character of the liquid. It can thus constitute an organic biological film on the surface, with a mineralizing power. The thickness of the film can vary to a large extent depending on its end use. A thick film can for example be obtained with the triple property: (i) sealing, (ii) self-supporting structure, (iii) intrinsic mineral contribution, while having the regenerating and healing character. This material, formed by an intimate association between organic matter and mineral matter, forms a film on any surface and crystallizes polymorphically depending on the condition and texture of the surface. The film thus obtained can be used on its support or be removed from the support for later use. The liquid can also be used later in the form of fibers. Any method known per se for forming a liquid into fibers can be used. This liquid has the advantage of being spinnable over a wide range of viscosities. In particular, a very simple stretching technique, in particular using tweezers, makes it possible to stretch fibers from the liquid. It is possible in this way to use it in situ (locally) or in another way, as a porous, invasive filling material, to fill as needed a given volume. The invasive nature of this material allows in particular biological materials, such as cells, to penetrate there and exert their effects in association with the properties of the shaped liquid. The same material can be used to bridge two distant tissue banks. The same principle can be applied to very large volumes, in particular by coating fibers of another chemical nature (for example carbon fibers). The fibers thus coated have a coating on their surface comprising the constituent elements of the liquid according to the invention and can be used to provide, for example, a healing interface.
Le fluide peut ainsi se présenter sous différentes formes, incluant liquide, films, revêtements et fibres. Selon la mise en forme désirée, il peut également se présenter sous forme d'un matériau poreux, notamment en utilisant, lors de sa mise en forme, un support poreux. Le matériau poreux peut être un film, un revêtement ou se présenter sous tout autre forme, tel qu'une forme sphérique.The fluid can thus come in different forms, including liquid, films, coatings and fibers. According to the desired shaping, it can also be in the form of a porous material, in particular by using, during its shaping, a porous support. The porous material may be a film, a coating or be in any other form, such as a spherical shape.
Le fluide selon l'invention comprend généralement un mélange des éléments constitutifs de la matière vivante, en particulier un mélange de protéines, de proteoglycanes, de polysaccharides, de lipides et de sels minéraux, ceci est plus particulièrement le cas pour un mollusque nacrier, avantageusement une huître du genre Pinctada (généralement prise dans son biotope naturel). La nature et la quantité de ces éléments peuvent varier dans une large mesure, notamment en fonction du mollusque, de son biotope, de la saison du prélèvement et des conditions du prélèvement.The fluid according to the invention generally comprises a mixture of the constituent elements of living matter, in particular a mixture of proteins, proteoglycans, polysaccharides, lipids and mineral salts, this is more particularly the case for a mother-of-pearl mollusc, advantageously an oyster of the genus Pinctada (generally taken in its natural biotope). The nature and quantity of these elements can vary to a large extent, particularly in function of the mollusc, its biotope, the season of the sample and the conditions of the sample.
Pour donner un ordre de grandeur, ces éléments ont été analysés dans un liquide donné (huître adulte du genre Pinctada prise dans son biotope naturel). La composition de ce liquide comprend de préférence au moins les composés suivants : des protéines en quantité moyenne de 0,5 mg/mL, des proteoglycanes en quantité moyenne de 3 mg/mL, des polysaccharides en quantité moyenne de 1,5 mg/mL, des lipides en quantité moyenne de 1 mg/mL et des sels minéraux en quantité moyenne de 7,5 mg/mL.To give an order of magnitude, these elements were analyzed in a given liquid (adult oyster of the genus Pinctada taken in its natural biotope). The composition of this liquid preferably comprises at least the following compounds: proteins in an average amount of 0.5 mg / mL, proteoglycans in an average amount of 3 mg / mL, polysaccharides in an average amount of 1.5 mg / mL , lipids in an average amount of 1 mg / mL and mineral salts in an average amount of 7.5 mg / mL.
Une analyse quantitative des oligo-éléments présents dans un liquide spécifique (identique à celui identifié ci-dessus) selon l'invention a été réalisée, en particulier en mesurant quantitativement les oligo-éléments présents par analyse par activation neutronique (AAN) et spectrométrie de masse couplée à une torche à plasma (ICP-MS). La composition de ce liquide (directement obtenu : sans déshydratation et modification ultérieure) comprend de préférence les oligoéléments choisis parmi : S, Mg, La, Zn, Br, Ce, Fe, Mn, Cl, Cu, K, Sr, Na et Ca. De préférence, les quantités de ces oligo-éléments sont respectivement les suivantes : 0,02 ; 2 ; 0,28 ; 0,4 ; 1 ,78 ; 5,5 ; 13,6 ; 50 ; 296 ; 143 ; 582 ; 1000 ; 5420 (exprimés en μg/g) et 38,8 (exprimés en μg/100g).A quantitative analysis of the trace elements present in a specific liquid (identical to that identified above) according to the invention was carried out, in particular by quantitatively measuring the trace elements present by analysis by neutron activation (ANA) and spectrometry of mass coupled to a plasma torch (ICP-MS). The composition of this liquid (directly obtained: without dehydration and subsequent modification) preferably comprises the trace elements chosen from: S, Mg, La, Zn, Br, Ce, Fe, Mn, Cl, Cu, K, Sr, Na and Ca Preferably, the amounts of these trace elements are respectively as follows: 0.02; 2; 0.28; 0.4; 1.78; 5.5; 13.6; 50; 296; 143; 582; 1000; 5420 (expressed in μg / g) and 38.8 (expressed in μg / 100g).
Une analyse quantitative des acides aminés présents sous forme libre ou lié dans un fluide selon l'invention a été réalisée, en particulier en mesurant quantitativement les acides aminés présents par l'analyseur d'acides aminés Beckmann 6300 utilisant la méthode classique à la ninhydrine. Le résultat de cette analyse est donné à la figure 1 (% en poids d'acides aminés par rapport au poids total d'acides aminés).A quantitative analysis of the amino acids present in free form or bound in a fluid according to the invention was carried out, in particular by quantitatively measuring the amino acids present by the amino acid analyzer Beckmann 6300 using the conventional method with ninhydrin. The result of this analysis is given in FIG. 1 (% by weight of amino acids relative to the total weight of amino acids).
La présente invention a aussi pour objet le fluide susceptible d'être obtenu par le procédé tel que décrit-avant. Le liquide (ou fluide) peut être non seulement le liquide obtenu directement après l'étape de récupération décrite ci-dessus, mais aussi celui obtenu après des étapes ultérieures non dénaturantes, telles que des étapes de filtration, de décontamination, de stérilisation, de congélation/décongélation, de modifications de viscosité, et/ou de mises en formes variées, telles que notamment celles identifiées ci-dessus.The present invention also relates to the fluid capable of being obtained by the process as described above. The liquid (or fluid) can be not only the liquid obtained directly after the recovery stage described above, but also that obtained after subsequent non-denaturing stages, such as stages of filtration, decontamination, sterilization, freezing / thawing, changes in viscosity, and / or various forms, such as in particular those identified above.
Plus particulièrement, le fluide selon l'invention est utilisé à titre de médicament.More particularly, the fluid according to the invention is used as a medicament.
La présente invention a également pour objet une composition, en particulier pharmaceutique, caractérisée en ce qu'elle comprend le fluide tel que défini ci- dessus, en particulier en association avec un excipient pharmaceutiquement acceptable.The present invention also relates to a composition, in particular pharmaceutical, characterized in that it comprises the fluid as defined above, in particular in combination with a pharmaceutically acceptable excipient.
Le liquide utilisé à titre de médicament ou dans la composition détaillée ci-dessous peut être celui obtenu directement après l'étape de récupération décrite ci-dessus, mais aussi celui obtenu après des étapes ultérieures de filtration, de décontamination, de stérilisation, de congélation/décongélation, de modifications de viscosité, et/ou de mises en formes variées, telles que notamment celles identifiées ci-dessus.The liquid used as a medicament or in the composition detailed below may be that obtained directly after the recovery stage described above, but also that obtained after subsequent stages of filtration, decontamination, sterilization, freezing / thawing, changes in viscosity, and / or various forms, such as in particular those identified above.
L'administration de la composition pharmaceutique ou du médicament selon l'invention peut être effectuée par voie topique, entérale ou parentérale. De préférence, la composition pharmaceutique est conditionnée sous une forme convenant à une application par voie topique, c'est à dire appliquée in situ à l'endroit où le tissu est lésé.The administration of the pharmaceutical composition or of the medicament according to the invention can be carried out by topical, enteral or parenteral route. Preferably, the pharmaceutical composition is packaged in a form suitable for topical application, that is to say applied in situ to the place where the tissue is injured.
Par voie entérale, la composition pharmaceutique peut se présenter sous forme de comprimés, de gélules, de dragées, de sirops, de suspensions, de solutions, de poudres, de granulés, d'émulsions, de microsphères ou nano-sphères ou vésicules lipidiques ou polymeriques permettant une libération contrôlée. Par voie parentérale, la composition peut se présenter sous forme de solutions ou suspensions pour perfusion ou pour injection.By enteral route, the pharmaceutical composition can be in the form of tablets, capsules, dragees, syrups, suspensions, solutions, powders, granules, emulsions, microspheres or nanospheres or vesicles lipidic or polymeric allowing controlled release. By parenteral route, the composition can be in the form of solutions or suspensions for infusion or for injection.
Selon un mode particulier de l'invention, la composition pharmaceutique est destinée à un usage local sur ou dans le tissu lésé. Ainsi, le support pharmaceutiquement acceptable est en particulier un excipient approprié à une application topique. Par voie topique, la composition pharmaceutique selon l'invention est plus particulièrement destinée au traitement des tissus mentionnés ci-dessus, en particulier au traitement de la peau et des os d'un mammifère, en particulier humain. Elle peut se présenter sous forme d'onguents, de crèmes, de laits, de pommades, de poudres, de tampons imbibés, de solutions, de gels, de sprays, de lotions ou de suspensions. Elle peut également se présenter sous forme de microsphères ou nano-sphères ou vésicules lipidiques ou polymeriques ou de patches polymeriques et d'hydrogels permettant une libération contrôlée. Cette composition par voie topique peut se présenter sous forme anhydre, sous forme aqueuse ou sous la forme d'une émulsion (émulsion eau/huile, huile/eau ou émulsion multiple).According to a particular embodiment of the invention, the pharmaceutical composition is intended for local use on or in the injured tissue. Thus, the pharmaceutically acceptable carrier is in particular an excipient suitable for topical application. By topical route, the pharmaceutical composition according to the invention is more particularly intended for the treatment of the tissues mentioned above, in particular for the treatment of the skin and bones of a mammal, in particular human. It can be in the form of ointments, creams, milks, ointments, powders, soaked pads, solutions, gels, sprays, lotions or suspensions. It can also be in the form of microspheres or nanospheres or lipid or polymeric vesicles or of polymeric patches and hydrogels allowing controlled release. This topical composition can be in anhydrous form, in aqueous form or in the form of an emulsion (water / oil, oil / water emulsion or multiple emulsion).
Le liquide selon l'invention est utilisé, de préférence par voie topique, à une concentration généralement comprise entre 0,02 % et 20 % en poids, de préférence entre 0,25 et 10 % en poids, et avantageusement entre 0,5 et 5 %, par rapport au poids total de la composition.The liquid according to the invention is used, preferably topically, at a concentration generally between 0.02% and 20% by weight, preferably between 0.25 and 10% by weight, and advantageously between 0.5 and 5%, relative to the total weight of the composition.
En particulier, la composition thérapeutique selon l'invention est caractérisée en ce qu'elle contient, en association avec des excipients inertes, une quantité thérapeutiquement efficace du liquide selon l'invention, notamment pour améliorer la cicatrisation cutanée, pour régénérer les tissus, ou pour favoriser l'ostéogenèse ou la minéralisation osseuse. Les tissus comprennent les tissus mous ou squelettiques, incluant en particulier le squelette, les os, le cartilage, les dents, ou tout autre tissu, en particulier la peau et ses phanères. La composition selon l'invention permet de traiter en particulier les désordres osseux, cartilagineux ou dentaires.In particular, the therapeutic composition according to the invention is characterized in that it contains, in association with inert excipients, a therapeutically effective amount of the liquid according to the invention, in particular for improving skin healing, for regenerating the tissues, or to promote osteogenesis or bone mineralization. Tissues include soft tissue or skeletal, including in particular the skeleton, bones, cartilage, teeth, or any other tissue, in particular the skin and its appendages. The composition according to the invention makes it possible to treat in particular bone, cartilaginous or dental disorders.
L'invention réside également dans une méthode pour améliorer la cicatrisation cutanée, pour régénérer les tissus, notamment ceux identifiés ci-dessus, ou pour favoriser l'ostéogenèse ou la minéralisation osseuse, comprenant l'administration à des sujets nécessitant d'un tel traitement d'une quantité thérapeutiquement efficace du liquide tel que défini précédemment.The invention also resides in a method for improving skin healing, for regenerating tissues, in particular those identified above, or for promoting osteogenesis or bone mineralization, comprising administration to subjects in need of such treatment a therapeutically effective amount of the liquid as defined above.
Dans le contexte de l'invention, le terme « traitement » désigne le traitement préventif, curatif, palliatif, ainsi que la prise en charge des patients (réduction de la douleur, amélioration de la qualité de vie, ralentissement de la progression du désordre, du traumatisme ou de la maladie), etc.In the context of the invention, the term “treatment” designates preventive, curative, palliative treatment, as well as the management of patients (reduction of pain, improvement of the quality of life, slowing down of the progression of the disorder, trauma or illness), etc.
La méthode ou le traitement peut en outre être réalisé en combinaison avec d'autres ingrédients ou traitements, tels que notamment d'autres composés actifs pour améliorer la cicatrisation cutanée, pour régénérer les tissus, ou pour favoriser l'ostéogenèse ou la minéralisation osseuse.The method or treatment can also be carried out in combination with other ingredients or treatments, such as in particular other active compounds to improve skin healing, to regenerate tissues, or to promote osteogenesis or bone mineralization.
Les compositions pharmaceutiques ou médicaments selon l'invention peuvent comprendre en outre au moins un autre ingrédient thérapeutiquement actif, pour une utilisation simultanée, séparée ou étalée dans le temps.The pharmaceutical compositions or medicaments according to the invention can also comprise at least one other therapeutically active ingredient, for simultaneous, separate or spread over time use.
Les compositions pharmaceutiques ou médicaments selon l'invention comprennent avantageusement un ou plusieurs excipients ou véhicules inertes, c'est à dire pharmaceutiquement inactifs et non toxiques. On peut citer par exemple des solutions salines, physiologiques, isotoniques, tamponnées, etc., compatibles avec un usage pharmaceutique et connues de l'homme du métier. Les compositions peuvent contenir un ou plusieurs agents ou véhicules choisis parmi les dispersants, solubilisants, stabilisants, conservateurs, etc. Des agents ou véhicules utilisables dans des formulations (liquides et/ou injectables et/ou solides) sont notamment la méthylcellulose, Phydroxyméthylcellulose, la carboxyméthylcellulose, les cylclodextrines, le polysorbate 80, le mannitol, la gélatine, le lactose, des huiles végétales ou animales, l'acacia, etc. Les compositions peuvent être formulées sous forme de suspension injectable, de gels, huiles, comprimés, suppositoires, poudres, gélules, capsules, etc., éventuellement au moyen de formes galéniques ou de dispositifs assurant une libération prolongée et/ou retardée. Pour ce type de formulation, on utilise avantageusement au moins un agent tel que la cellulose, des carbonates ou des amidons.The pharmaceutical compositions or medicaments according to the invention advantageously comprise one or more inert excipients or vehicles, that is to say pharmaceutically inactive and non-toxic. Mention may be made, for example, of saline, physiological, isotonic, buffered solutions, etc., compatible with pharmaceutical use and known to those skilled in the art. The essays may contain one or more agents or vehicles chosen from dispersants, solubilizers, stabilizers, preservatives, etc. Agents or vehicles usable in formulations (liquids and / or injectables and / or solids) are in particular methylcellulose, Phydroxymethylcellulose, carboxymethylcellulose, cylclodextrins, polysorbate 80, mannitol, gelatin, lactose, vegetable or animal oils , acacia, etc. The compositions can be formulated in the form of an injectable suspension, gels, oils, tablets, suppositories, powders, capsules, capsules, etc., optionally by means of dosage forms or devices ensuring sustained and / or delayed release. For this type of formulation, advantageously at least one agent such as cellulose, carbonates or starches is used.
Les compositions telles que décrites précédemment peuvent bien entendu en outre contenir des additifs pharmacodynamiquement actifs ou une combinaison de ces additifs, et notamment : des agents mouillants, des émollients, des agents hydratants, comme le glycérol, le PEG 400 ou bien encore l'urée.The compositions as described above can of course also contain pharmacodynamically active additives or a combination of these additives, and in particular: wetting agents, emollients, hydrating agents, such as glycerol, PEG 400 or else urea .
Ces compositions peuvent également contenir des agents d'amélioration de la saveur, des agents conservateurs tels que les esters de l'acide parahydroxybenzoïque, les agents stabilisants, des agents régulateurs d'humidité, des agents régulateurs de pH, des agents modificateurs de pression osmotique, des agents émulsionnants, des antioxydants, tels que l'α-tocophérol, le butylhydroxyanisole ou le butylhydroxytoluène.These compositions may also contain flavor enhancers, preservatives such as parahydroxybenzoic acid esters, stabilizers, moisture regulators, pH regulators, osmotic pressure modifiers , emulsifying agents, antioxidants, such as α-tocopherol, butylhydroxyanisole or butylhydroxytoluene.
Bien entendu, l'homme du métier veillera à choisir le ou les éventuels composés à ajouter à ces compositions de telle manière que les propriétés avantageuses attachées intrinsèquement à la présente invention ne soient pas ou substantiellement pas altérées par l'addition envisagée.Of course, those skilled in the art will take care to choose the optional compound (s) to be added to these compositions in such a way that the advantageous properties intrinsically attached to the present invention are not or substantially not altered by the addition envisaged.
L'administration peut être réalisée par toute méthode connue de l'homme du métier, notamment par voie orale, topique ou par injection, typiquement par voie intra- péritonéale, intra-cérébrale, intra-thécale, intra-veineuse, intra-artérielle ou intramusculaire. L'administration par voie topique ou orale est préférée. S'agissant d'un traitement à long terme et non topique, la voie d'administration préférée sera sublinguale, orale ou transcutanée.The administration can be carried out by any method known to those skilled in the art, in particular by the oral, topical or injection route, typically by the intraperitoneal, intra-cerebral, intra-thecal, intravenous, intra-arterial or intramuscular route. Topical or oral administration is preferred. For long-term, non-topical treatment, the preferred route of administration will be sublingual, oral or transcutaneous.
Pour les injections, les composés sont généralement conditionnés sous forme de suspensions liquides, qui peuvent être injectées au moyen de seringues ou de perfusions, par exemple. Il est entendu que le débit et/ou la dose injectée, ou de manière générale la dose à administrer, peuvent être adaptés par l'homme du métier en fonction du sujet à traiter, du désordre, du mode d'administration, etc.. Il est entendu que des administrations répétées peuvent être réalisées, éventuellement en combinaison avec d'autres ingrédients actifs ou tout véhicule acceptable sur le plan pharmaceutique (tampons, solutions saline, isotonique, en présence d'agents stabilisants, etc.).For injections, the compounds are generally packaged in the form of liquid suspensions, which can be injected using syringes or infusions, for example. It is understood that the flow rate and / or the dose injected, or in general the dose to be administered, can be adapted by a person skilled in the art according to the subject to be treated, the disorder, the mode of administration, etc. It is understood that repeated administrations can be carried out, optionally in combination with other active ingredients or any pharmaceutically acceptable vehicle (buffers, saline, isotonic solutions, in the presence of stabilizing agents, etc.).
Selon un aspect particulier, l'invention concerne un dispositif, plus spécifiquement adapté à une injection sous-cutanée ou percutanée, comprenant le liquide tel que défini ci-avant et un excipient ou support physiologiquement acceptable. Ce dispositif peut notamment être sous forme de seringues ou de perfusions.According to a particular aspect, the invention relates to a device, more specifically suitable for subcutaneous or percutaneous injection, comprising the liquid as defined above and a physiologically acceptable excipient or support. This device can in particular be in the form of syringes or infusions.
Selon une variante, le dispositif médical de la présente invention peut être destiné à être implanté dans ou sur le corps d'un mammifère, le liquide tel que défini ci-avant pouvant se situer à l'intérieur d'une membrane adaptée à cette usage.According to a variant, the medical device of the present invention may be intended to be implanted in or on the body of a mammal, the liquid as defined above may be located inside a membrane suitable for this use .
Selon un autre aspect particulier, l'invention concerne un complément alimentaire comprenant le liquide tel que défini ci-avant. Ce complément alimentaire est en particulier destiné à améliorer la cicatrisation cutanée, à régénérer les tissus, ou à favoriser l'ostéogenèse ou la minéralisation des os ou de la denture.According to another particular aspect, the invention relates to a food supplement comprising the liquid as defined above. This food supplement is in particular intended to improve cutaneous healing, to regenerate tissues, or to favor osteogenesis or mineralization of bones or teeth.
L'invention est utilisable chez les mammifères, en particulier chez l'être humain. D'autres aspects et avantages de la présente invention apparaîtront à la lecture des exemples qui suivent, qui doivent être considérés comme illustratifs et non limitatifs.The invention can be used in mammals, in particular in humans. Other aspects and advantages of the present invention will appear on reading the examples which follow, which should be considered as illustrative and not limiting.
EXEMPLESEXAMPLES
Exemple 1. Récupération du liquide à partir d'un animal sacrifié Le mollusque est sacrifié de manière à ce que le liquide reste dans la coquille. On décolle avec précaution le manteau ; on utilise ensuite une réaction enzymatique pour décoller complètement le liquide de la coquille. La réaction est ensuite stoppée par une anti-protéase. Le liquide est ensuite prélevé à la surface de la coquille. Aussitôt prélevé, le liquide peut être congelé. Il peut être stérilisé par filtration sur millipore 0, 20μm et être lyophilisé tout en conservant ses propriétés.Example 1. Recovery of the liquid from a sacrificed animal The mollusk is sacrificed so that the liquid remains in the shell. We take off the coat carefully; an enzymatic reaction is then used to completely detach the liquid from the shell. The reaction is then stopped by an anti-protease. The liquid is then taken from the surface of the shell. As soon as collected, the liquid can be frozen. It can be sterilized by filtration on a 0.20μm millipore and can be freeze-dried while retaining its properties.
Exemple 2. Récupération du liquide à partir d'un animal vivantExample 2. Recovery of the liquid from a living animal
Une huître de l'espèce de Pinctada margaritifera est entre-ouverte grâce à un ecarteur suivant les pratiques bien connues de l'homme de l'art en perliculture. L'eau en excès encore présente est évacuée. L'animal est disposé de façon horizontale. Le manteau est délicatement séparé de la coquille inférieure à l'aide d'une pince, puis le liquide est prélevé à l'aide d'une seringue, par exemple.An oyster of the species of Pinctada margaritifera is half-opened thanks to a retractor according to the practices well known to those skilled in the art in pearl farming. The excess water still present is discharged. The animal is arranged horizontally. The mantle is delicately separated from the lower shell using forceps, then the liquid is withdrawn using a syringe, for example.
Pour le groupe 1 : taille moyenne: 77mm épaisseur moyenne : 26,12mm, on obtient 200 à 300 μL de liquide prélevé. Pour le groupe 2 : taille moyenne: 85,2mm épaisseur moyenne : 25,6mm, on obtient 300 à 500 μL de liquide prélevé.For group 1: average size: 77mm average thickness: 26.12mm, we obtain 200 to 300 μL of liquid sampled. For group 2: average size: 85.2mm average thickness: 25.6mm, we obtain 300 to 500 μL of liquid sampled.
Pour le groupe 3 : taille moyenne: 104mm épaisseur moyenne : 36,9mm, on obtient 500 à 800 μL de liquide prélevé. Pour le groupe 4 : taille moyenne: 108,6mm épaisseur moyenne : 36mm, on obtient 800 à 1000 μL de liquide prélevé.For group 3: average size: 104mm average thickness: 36.9mm, we obtain 500 to 800 μL of liquid sampled. For group 4: average size: 108.6mm average thickness: 36mm, we obtain 800 to 1000 μL of liquid sampled.
Le liquide est : - filtré - lyophilisé - quelques semaines plus tard, le liquide est reformé avec de l'eau stérile - il est étalé sous forme de film épais.The liquid is: - filtered - lyophilized - a few weeks later, the liquid is reformed with sterile water - it is spread in the form of a thick film.
Exemple 3. Récupération du liquide à partir d'un mollusque non nacrierExample 3. Recovery of the Liquid from a Non-Pearl Shellfish
Une huître de l'espèce Ostrea edulis est sacrifiée. Sa coquille est ouverte par cisaillement du muscle sans séparer les valves (par exemple à 60° l'une de l'autre).An oyster of the species Ostrea edulis is sacrificed. Its shell is opened by shearing the muscle without separating the valves (for example at 60 ° from one another).
L'huître est fixée et le manteau est décollé. De cette manière, il est possible de récolter le liquide de la première valve. L'animal est ensuite retourné pour opérer sur l'autre face. Dans ce cas, une quantité de 50 à 100μl par valve est obtenue.The oyster is fixed and the coat is peeled off. In this way, it is possible to collect the liquid from the first valve. The animal is then turned over to operate on the other side. In this case, an amount of 50 to 100μl per valve is obtained.
Le liquide récolté sur une population d'une vingtaine d'animaux subit un conditionnement similaire à celui de l'exemple 2.The liquid collected from a population of around twenty animals is subjected to conditioning similar to that of Example 2.
Le liquide est : - filtré - lyophilisé - quelques semaines plus tard, le liquide est reformé avec de l'eau stérile - il est étalé sous forme de film épaisThe liquid is: - filtered - lyophilized - a few weeks later, the liquid is reformed with sterile water - it is spread in the form of a thick film
On constate que le liquide se reconstitue même après lyophilisation, qu'il forme un film sur un support de polypropylene. Ce film se solidifie par auto-organisation et auto-cicatrisation.It is found that the liquid is reconstituted even after lyophilization, that it forms a film on a polypropylene support. This film solidifies by self-organization and self-healing.
L'examen au microscope optique montre qu'il est composé d'une fraction organique et d'une fraction minérale. Examination under an optical microscope shows that it is composed of an organic fraction and a mineral fraction.

Claims

REVENDICATIONS
1. Procédé d'obtention d'un fluide extrapalleal, caractérisé en ce qu'il comprend la récupération du fluide extrapalleal se situant entre la coquille et le corps d'un mollusque, en particulier le manteau dudit mollusque.1. Method for obtaining an extrapalleal fluid, characterized in that it comprises recovering the extrapalleal fluid located between the shell and the body of a mollusk, in particular the mantle of said mollusk.
2. Procédé selon la revendication 1, caractérisé en ce que le mollusque à partir duquel est récupéré le fluide est un mollusque bivalve, avantageusement nacrier.2. Method according to claim 1, characterized in that the mollusc from which the fluid is recovered is a bivalve mollusc, advantageously pearly.
3. Procédé selon la revendication 2, caractérisé en ce que le mollusque est choisi parmi des huîtres et plus spécifiquement des huîtres du genre Pinctada et plus particulièrement de l'espèce margaritifera ou maxima, du genre Ostrea, en particulier de l'espèce edulis, ou du genre Crassostrea, en particulier de l'espèce gigas.3. Method according to claim 2, characterized in that the mollusc is chosen from oysters and more specifically oysters of the genus Pinctada and more particularly of the species margaritifera or maxima, of the genus Ostrea, in particular of the species edulis, or of the genus Crassostrea, in particular of the species gigas.
4. Procédé selon la revendication 1, caractérisé en ce que le mollusque à partir duquel est récupéré le fluide est un céphalopode, comme par exemple le nautile, ou encore un gastéropode, comme par exemple le bigorneau.4. Method according to claim 1, characterized in that the mollusc from which the fluid is recovered is a cephalopod, such as the nautilus, or a gastropod, such as the periwinkle.
5. Procédé selon l'une des revendications précédentes, caractérisé en ce que, préalablement à la récupération, le manteau du mollusque est décollé en tout ou partie.5. Method according to one of the preceding claims, characterized in that, before recovery, the shell of the mollusc is detached in whole or in part.
6. Procédé selon l'une des revendications précédentes, caractérisé en ce que, préalablement à la récupération, lorsque le mollusque est bi-valve, au moins une des deux valves est soulevée de manière à pouvoir accéder au corps du mollusque. 6. Method according to one of the preceding claims, characterized in that, prior to recovery, when the mollusk is bi-valve, at least one of the two valves is raised so as to be able to access the body of the mollusk.
7. Procédé selon l'une des revendications précédentes, caractérisé en ce que, préalablement à la récupération, le décollement du fluide extrapalleal de la coquille est réalisé manuellement, généralement à l'aide d'un outil approprié.7. Method according to one of the preceding claims, characterized in that, prior to recovery, detachment of the extrapalleal fluid from the shell is carried out manually, generally using a suitable tool.
8. Procédé selon l'une des revendications précédentes, caractérisé en ce que le fluide extrapalleal est récupéré à la surface de la coquille à l'aide d'un instrument approprié, tel que notamment racloir, spatule ou seringue.8. Method according to one of the preceding claims, characterized in that the extrapalleal fluid is recovered on the surface of the shell using an appropriate instrument, such as in particular a scraper, spatula or syringe.
9. Procédé selon l'une des revendications précédentes, caractérisé en ce qu'il comprend en outre des étapes non dénaturantes du fluide ainsi récupéré, en particulier une ou des étapes de filtration, de décontamination, de stérilisation, de congélation/décongélation, de modifications de viscosité, et/ou de mises en formes variées.9. Method according to one of the preceding claims, characterized in that it further comprises non-denaturing steps of the fluid thus recovered, in particular one or more steps of filtration, decontamination, sterilization, freezing / thawing, viscosity changes, and / or various forms.
10. Procédé selon l'une des revendications précédentes, caractérisé en ce que le fluide récupéré est stérilisé ou décontaminé.10. Method according to one of the preceding claims, characterized in that the recovered fluid is sterilized or decontaminated.
11. Procédé selon l'une des revendications précédentes, caractérisé en ce que le fluide récupéré est déshydraté, notamment par lyophilisation.11. Method according to one of the preceding claims, characterized in that the recovered fluid is dehydrated, in particular by lyophilization.
12. Procédé selon l'une des revendications précédentes, caractérisé en ce que le fluide récupéré est congelé, de préférence juste après la récupération ou après filtration.12. Method according to one of the preceding claims, characterized in that the recovered fluid is frozen, preferably just after recovery or after filtration.
13. Procédé selon l'une quelconque des revendications précédentes, caractérisé en ce que le fluide récupéré est mis sous forme de film ou de revêtement.13. Method according to any one of the preceding claims, characterized in that the recovered fluid is put in the form of film or coating.
14. Procédé selon l'une quelconque des revendications 1 à 12, caractérisé en ce que le fluide récupéré est mis sous forme de fibres. 14. Method according to any one of claims 1 to 12, characterized in that the recovered fluid is put in the form of fibers.
15. Procédé selon l'une quelconque des revendications 1 à 12, caractérisé en ce que le fluide récupéré est mis sous forme d'un matériau poreux.15. Method according to any one of claims 1 to 12, characterized in that the recovered fluid is put in the form of a porous material.
16. Fluide susceptible d'être obtenu par le procédé tel que défini dans l'une quelconque des revendications précédentes.16. Fluid capable of being obtained by the process as defined in any one of the preceding claims.
17. Composition pharmaceutique, caractérisée . en ce qu'elle comprend le fluide défini dans la revendication précédente et un excipient pharmaceutiquement acceptable.17. Pharmaceutical composition, characterized. in that it comprises the fluid defined in the preceding claim and a pharmaceutically acceptable excipient.
18. Composition selon la revendication précédente, destinée à améliorer la cicatrisation cutanée, à régénérer les tissus mous ou squelettiques, ou à favoriser l'ostéogenèse ou la minéralisation osseuse.18. Composition according to the preceding claim, intended to improve skin healing, to regenerate soft or skeletal tissues, or to promote osteogenesis or bone mineralization.
19. Composition selon la revendication 17, destinée au traitement des désordres osseux, cartilagineux ou dentaires.19. Composition according to claim 17, intended for the treatment of bone, cartilaginous or dental disorders.
20. Composition pharmaceutique selon l'une des revendications précédentes 17 à 19, caractérisée en ce que le support pharmaceutiquement acceptable est un excipient approprié à une application topique, entérale ou parentérale.20. Pharmaceutical composition according to one of the preceding claims 17 to 19, characterized in that the pharmaceutically acceptable carrier is an excipient suitable for topical, enteral or parenteral application.
21. Composition selon la revendication précédente, caractérisée en ce que le support pharmaceutiquement acceptable est un excipient approprié à une application topique.21. Composition according to the preceding claim, characterized in that the pharmaceutically acceptable carrier is an excipient suitable for topical application.
22. Composition selon l'une quelconque des revendications précédentes 17 à 21 , caractérisée en ce qu'elle comprend en outre au moins un autre ingrédient thérapeutiquement actif pour une utilisation simultanée, séparée ou étalée dans le temps. 22. Composition according to any one of the preceding claims 17 to 21, characterized in that it also comprises at least one other therapeutically active ingredient for simultaneous, separate or spread over time use.
23. Fluide selon la revendication 16, à titre de médicament.23. Fluid according to claim 16, as a medicament.
24. Dispositif comprenant le fluide tel que défini dans la revendication 16 et un excipient ou support physiologiquement acceptable.24. Device comprising the fluid as defined in claim 16 and a physiologically acceptable excipient or support.
25. Dispositif selon la revendication précédente, caractérisé en ce que sa forme est adaptée à une injection sous-cutanée ou percutanée, en particulier sous. forme de seringue ou de perfusion.25. Device according to the preceding claim, characterized in that its shape is suitable for subcutaneous or percutaneous injection, in particular under. syringe or infusion.
26. Dispositif selon la revendication 24, caractérisé en ce que le fluide est à l'intérieur d'une membrane.26. Device according to claim 24, characterized in that the fluid is inside a membrane.
27. Complément alimentaire comprenant le fluide tel que défini à la revendication 16. 27. Food supplement comprising the fluid as defined in claim 16.
EP05717633A 2004-02-13 2005-02-14 Extrapallial fluid of molluscs, method of obtaining same, formulation and use thereof Withdrawn EP1722803A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FR0401474A FR2866238B1 (en) 2004-02-13 2004-02-13 EXTRAPALLEM LIQUIDS OF MOLLUSCS, PROCESSING, FORMULATION AND USE
PCT/FR2005/000344 WO2005077388A1 (en) 2004-02-13 2005-02-14 Extrapallial fluid of molluscs, method of obtaining same, formulation and use thereof

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EP1722803A1 true EP1722803A1 (en) 2006-11-22

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EP05717633A Withdrawn EP1722803A1 (en) 2004-02-13 2005-02-14 Extrapallial fluid of molluscs, method of obtaining same, formulation and use thereof

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US (1) US20070128293A1 (en)
EP (1) EP1722803A1 (en)
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WO (1) WO2005077388A1 (en)

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Publication number Priority date Publication date Assignee Title
US20100233111A1 (en) * 2008-04-06 2010-09-16 William Wang Gastropod biological fluid, method of making and refining and use
JP5768054B2 (en) 2009-10-28 2015-08-26 クリアウォーター シーフーズ リミテッド パートナーシップ Mollusk processing apparatus and related methods
US8205623B2 (en) * 2010-04-05 2012-06-26 Rovcal, Inc. Coatings for personal grooming apparatus containing calcium carbonate derived from mollusk shells or pearls

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FR2502007A1 (en) * 1981-03-18 1982-09-24 Simonin Renee Products contg. mollusc eggs prior to deposition - having cicatrising, revitalising and rejuvenating effects
FR2595247B1 (en) * 1986-03-06 1988-09-23 Sederma Sa USE IN COSMETOLOGY OF MUCUS EXTRACTS OR DIGESTIVE SUGAS OF GASTEROPODS AND PREPARATION OF SUCH EXTRACTS
FR2713482B1 (en) * 1993-12-10 1996-03-01 Georges Camprasse Injectable implant for the correction of wrinkles and dermal depressions.
WO1996035786A1 (en) * 1995-05-09 1996-11-14 Fujikawa, Yoshio Pearl protein (nacrein) and process for producing the same
FR2743075B1 (en) * 1995-12-28 1998-03-27 Centre Nat Rech Scient PROCESS FOR THE PREPARATION OF ACTIVE SUBSTANCES FROM NACRE, PRODUCTS OBTAINED, USEFUL IN PARTICULAR AS MEDICAMENTS
JPH1014438A (en) * 1996-07-01 1998-01-20 Tokuzo Hirose Pearl and its production
US20030027258A1 (en) * 2001-08-02 2003-02-06 Chang Fang-Tseh Frank Methods and compositions for pearl oyster cultivation
US7404378B2 (en) * 2006-02-17 2008-07-29 Batzer William B Pearl culture method and product

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See references of WO2005077388A1 *

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US20070128293A1 (en) 2007-06-07
FR2866238B1 (en) 2006-12-15
WO2005077388A1 (en) 2005-08-25
FR2866238A1 (en) 2005-08-19

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