Classifications

• • C—CHEMISTRY; METALLURGY
  • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
  • C09J—ADHESIVES; NON-MECHANICAL ASPECTS OF ADHESIVE PROCESSES IN GENERAL; ADHESIVE PROCESSES NOT PROVIDED FOR ELSEWHERE; USE OF MATERIALS AS ADHESIVES
  • C09J175/00—Adhesives based on polyureas or polyurethanes; Adhesives based on derivatives of such polymers
  • C09J175/04—Polyurethanes
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
  • A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
  • A61L24/043—Mixtures of macromolecular materials
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
  • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/04—Antibacterial agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P41/00—Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
  • A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
  • A61P9/14—Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
  • C08G18/00—Polymeric products of isocyanates or isothiocyanates
  • C08G18/06—Polymeric products of isocyanates or isothiocyanates with compounds having active hydrogen
  • C08G18/08—Processes
  • C08G18/10—Prepolymer processes involving reaction of isocyanates or isothiocyanates with compounds having active hydrogen in a first reaction step
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
  • C08G18/00—Polymeric products of isocyanates or isothiocyanates
  • C08G18/06—Polymeric products of isocyanates or isothiocyanates with compounds having active hydrogen
  • C08G18/28—Polymeric products of isocyanates or isothiocyanates with compounds having active hydrogen characterised by the compounds used containing active hydrogen
  • C08G18/40—High-molecular-weight compounds
  • C08G18/42—Polycondensates having carboxylic or carbonic ester groups in the main chain
  • C08G18/4266—Polycondensates having carboxylic or carbonic ester groups in the main chain prepared from hydroxycarboxylic acids and/or lactones
  • C08G18/428—Lactides
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
  • C08G18/00—Polymeric products of isocyanates or isothiocyanates
  • C08G18/06—Polymeric products of isocyanates or isothiocyanates with compounds having active hydrogen
  • C08G18/28—Polymeric products of isocyanates or isothiocyanates with compounds having active hydrogen characterised by the compounds used containing active hydrogen
  • C08G18/40—High-molecular-weight compounds
  • C08G18/64—Macromolecular compounds not provided for by groups C08G18/42 - C08G18/63
  • C08G18/6415—Macromolecular compounds not provided for by groups C08G18/42 - C08G18/63 having nitrogen
  • C08G18/6446—Proteins and derivatives thereof
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
  • C08L89/00—Compositions of proteins; Compositions of derivatives thereof
• • C—CHEMISTRY; METALLURGY
  • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
  • C09J—ADHESIVES; NON-MECHANICAL ASPECTS OF ADHESIVE PROCESSES IN GENERAL; ADHESIVE PROCESSES NOT PROVIDED FOR ELSEWHERE; USE OF MATERIALS AS ADHESIVES
  • C09J189/00—Adhesives based on proteins; Adhesives based on derivatives thereof
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L2430/00—Materials or treatment for tissue regeneration
  • A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
  • Y10T428/00—Stock material or miscellaneous articles
  • Y10T428/29—Coated or structually defined flake, particle, cell, strand, strand portion, rod, filament, macroscopic fiber or mass thereof
  • Y10T428/2982—Particulate matter [e.g., sphere, flake, etc.]

Definitions

• the invention relates to combination of individual components of peptides, polyphenol oxidases and substituted polyhydroxy aromatics, their use J for modifying surfaces or for fixing reagents to surfaces and in the aqueous environment can be used adhesive bonding methods.
• MAPs muscle adhesive proteins
• Adhesives based on MAP are known in the art which can be used in an aqueous environment. MAP are thereby, e.g. linked directly by catechol oxidase
• EP 0 947 142 discloses increasing the molecular weight of proteins by their crosslinking by multi-copper enzymes, e.g. Laccases. The described increase in the molecular weight of the substrate proteins occurred after 17 hours of incubation.
• the proteins crosslinked by means of the method according to EP 0 947 142 are particularly suitable for use in foods, e.g. to change the consistency of sausage.
• crosslinking of proteins by substituted dihydroxyaromatics is not disclosed in EP 0 947 142.
• the object is to provide a combination which is particularly suitable both for bonding in an aqueous environment and for the functionalization of surfaces.
• Still difficult today is the acute supply of severely bleeding wounds. Here is the danger of bleeding in the foreground. If wound adhesives are to be used, there is a risk that adhesive components will be flushed away with the bloodstream and the bonding occurs at an undesirable location. This greatly increases the risk of thrombosis.
• a combination with a hemostatic mineral component is expedient.
• the synthesis products used hitherto for hemostasis are alkali and alkaline earth aluminum silicates of different composition.
• Synthetic lithium aluminosilicates (EP 1176991 A1, WO 00/69480) have been developed especially for wound care and are sold under the trade name CERDAC.
• CERDAC Synthetic lithium aluminosilicates
• this object has been achieved by various combinations of components - according to the features of the claims. Adhesive times in the range of minutes or less and an acceptable bond strength, e.g. allow use as a tissue adhesive.
• the combinations according to the invention are furthermore particularly suitable for the modification and / or coating of surfaces.
• the invention provides a combination comprising
• the components are preferably brought into contact simultaneously with a substrate carrying amino groups or functionalized with amino groups.
• the amino groups can be supported on surfaces, e.g.
• Implants according to methods known in the art, e.g.
• Plasma processes (Schröder, et al., Improved low- pressure raicrowave plasma assisted amino functionalization of polymers, plasma processes and polymers, Weinheim, Germany, Weinheim: Wiley-VCH, 2005; Schröder et al. , Plasma-Induced Surface Functionalization of Polymeric Biomaterials in Ammonia Plasma, Contrib. Plasma Phys. 41, 2001, 562-572; Meyer-Plath et al, Current trends in biomaterial surface functionalization - nitrogen-containing plasma assisted processes with enhanced selectivity, Vacuum 71, 2003, 391-406).
• Substrates or surfaces used in the methods of the invention may be e.g. have a surface containing at least about 2% free amino groups.
• the term surface describes not only flat surfaces, but boundary layers of a solid substrate in any shape. Substrates can thus e.g. also be beads or complex shaped surfaces.
• polyhydroxyaromatics can be applied to surfaces bearing free amino groups or functionalized with free amino groups.
• Polyhydroxyaromatics are aromatic compounds having more than one hydroxyl group attached to the aromatic ring or rings.
• the polyhydroxyaromatic is not itself a peptide component, but preferably a small organic molecule.
• the polyhydroxyaromatic may be a monocyclic dihydroxyaromatic, a bicyclic dihydroxyaromatic, a polycyclic dihydroxyaromatic, a bicyclic trihydroxyaromatic or a polycyclic trihydroxyaromatic. It preferably has a molecular weight of 110-1,100 g / mol.
• the polyhydroxyaromatic is a substituted monocyclic dihydroxyaromatic.
• the peptides are in the Polymerization achieved particularly favorable properties of the combination. Different substitutions can be used to achieve special properties adapted to the application.
• the substituted dihydroxyaromatic serves as a substrate of polyphenol oxidases, in particular laccases.
• Substituted means in the context of this invention that residues are bound to one molecule, in this case at the aromatic next to the two hydroxyl groups still 1, 2, 3 or 4 further radicals, one of these independently one or more radicals H, CH 3 , COH, COCH 3 , CONH 2 , CON-alkyl, CON-alkyl-OH, COOH, COO-alkyl, alkyl or may be a substituted aromatic, but not all radicals are H.
• the aromatic may also be part of a bicyclic or polycyclic aromatic ring system. With a substituted dihydroxyaromatic, 2 of the radicals H are necessary as available binding sites.
• This method can be used to modify surfaces, e.g. for modifying implants, surgical instruments, such as endoscopy devices, or vessels for cell culture or diagnostics.
• biofilms By modified by loading with polyphenols surface, the formation of biofilms can be prevented. This solves a major problem, especially in endoscopy.
• an in vivo or in vitro method of adhesion in which the components of the combination according to the invention or of the medical device are brought into contact with each other and with at least two substrates to be joined, which have hydroxyl groups, mercapto groups and / or amino groups.
• At least one of the substrates is a tissue, in particular an organ, connective tissue, skin, tendon, blood vessel and / or nerve (soft tissue) and / or tooth or tooth compartment (enamel, dentin and / or cement) and / or bone
• a tissue in particular an organ, connective tissue, skin, tendon, blood vessel and / or nerve (soft tissue) and / or tooth or tooth compartment (enamel, dentin and / or cement) and / or bone
• a particularly advantageous application is the fixation of drug-delivery systems in the tissue, the first step being the generation of amino groups on the system surface, preferably with plasma-assisted methods, for example, polydimethylsiloxane in an argon microwave plasma with an admixture of 1%. Ethyendiamin plasma-treated at a pressure of 0.5 mbar at a power of 500 W.
• the fixation in the tissue by means of polyphenols and polyphenol oxidase can therefore also be spheres, for example (beads) or complex shaped surfaces that are functionalized with amino groups.
• cytostatic drug delivery systems can be fixed in the environment of a tumor in order to reduce the side effect of systemic tumor therapy.
• the substrates are wound edges which are to be bonded.
• it may also be e.g. act to bond together inorganic or organic substrates that are not tissue, such as plastic or ceramic surfaces.
• a medical application is e.g. conceivable in the bonding of implants. Non-medical applications are found in the bonding of substrates under water or in an aqueous environment.
• substrates not only macroscopic substrates, but also other reagents containing amino groups (and / or hydroxyl groups), such as peptides, other drugs and / or cells. These can each other, but also be connected to solid carriers.
• reagents containing amino groups (and / or hydroxyl groups) such as peptides, other drugs and / or cells.
• These can each other, but also be connected to solid carriers.
• the use of the combination according to the invention for the binding of peptides and / or reagents which contain amino groups and / or hydroxyl groups, and / or cells to surfaces which contain amino groups and / or hydroxyl groups or are functionalized with amino groups and / or hydroxyl groups is made possible.
• Such reagents may be, for example, diagnostics, medicinal agents (e.g., antibiotics, hormones, immune system affecting substances, cytotoxic agents), antibodies, antigens or other proteins.
• diagnostics e.g. Diagnostics with a covalent link between a carrier, e.g. an ELISA plate, and a diagnostically useful peptide.
• a carrier e.g. an ELISA plate
• diagnostically useful peptide e.g. Diagnostics with a covalent link between a carrier, e.g. an ELISA plate, and a diagnostically useful peptide.
• a carrier e.g. an ELISA plate
• diagnostically useful peptide e.g. Diagnostics with a covalent link between a carrier, e.g. an ELISA plate, and a diagnostically useful peptide.
• the sensitivity of detection reactions is improved. As more intensive rinses can be used, the specificity of the detection reaction is increased.
• surfaces containing reactive groups can be
• connection of the substrates takes place by crosslinking of hydroxyl groups and / or amino groups of the oligopeptides and the substrates by the dihydroxyaromatics.
• R 1 and / or R 2 independently of one another are H, CH 3 , COH, COCH 3 , CONH 2 , CON-alkyl, CON-alkyl-OH, COOH, COO-alkyl, alkyl or substituted aromatic, in particular H, CH 3 , Alkyl or substituted aromatic, not both are H.
• the components R4-NH 2 (peptide, and / or protein and / or amino group-containing reagent and / or cell, in particular amino acid, biological active substance (antibiotic, drugs, diagnostic reagent) are via the bridging molecule to an amino-functionalized surface / matrix R3 -NH 2 is bonded.
• the reaction can proceed at about 2-80 0 C, preferably, however, a temperature of about 10-60 0 C, or about 20-37 0 C or 25-3O 0 C. This means that all procedures is The particularly gentle reaction conditions allow the fixation of even very sensitive biological agents.
• the invention likewise relates to a bonding process carried out in an aqueous medium using the combination according to the invention described above, e.g. a method of bonding under water.
• the processes of the invention are preferably carried out with one or more aqueous solvents, but the consistency of the components of the combination used need not be liquid, but a pasty consistency is also suitable.
• the pH is preferably 2-10, especially 4-8 or 7.
• the invention also relates to products obtainable by the process according to the invention. In this product, two substrates can be crosslinked by the adhesive according to the invention.
• peptides and / or proteins and / or amino-containing reagents and / or cells are bound to a surface
• diagnostic agents eg antibodies or antigens
• medical agents eg antibiotics, cytostatics , Hormones, substances affecting the immune system
• / or other proteins bound to a surface eg plastic surfaces, surfaces of cell culture plates, ELISA plates or other vessels for diagnostic applications or surfaces of implants.
• the surface is modified by bridging molecules of the combination according to the invention, ie the surface is covalently bonded to the polymerized components of the combination according to the invention and optionally other reagents.
• the combination according to the invention comprises the individual components a) oligopeptides containing at least one diamino acid, preferably at least two diamino acids, and b) substituted dihydroxyaromatics or substrates of polyphenol oxidases (preferably of laccases) and c) polyphenol oxidases (preferably laccases).
• the polymer is preferably a peptide.
• the peptide used is both an oligopeptide (2 to about 100 amino acids in length, preferably about 4 to about 20 amino acids in length or about 6 to about 10 amino acids in length) and a protein (protein, about 100 to about 5,000 Amino acids length, preferably 100-1000 or 100-200 amino acids in length).
• the peptide has a length of 10-1000 amino acids.
• the peptide may have a molecular weight of about 1 to about 100 or about 200 kDa, more preferably about 2 to about 50 kDa, or about 5 to about 20 kDa. It may be modified or substituted, eg glycosylated.
• modified or atypical amino acids such as hydroxyoxy may also be present in the oligopeptide.
• D-amino acids instead of or in addition to L-amino acids is possible and slows down the degradation of the peptide.
• the amino groups may be primary or secondary amino groups. However, especially reactive are primary amino groups. At least one of the reactive groups of the peptide is preferably part of a diamino acid, e.g. from lysine.
• the peptide therefore preferably comprises at least one diamino acid, preferably at least 2, 3, 4, 5 or more diamino acids. Preference is given to using lysine-containing peptides (or oligopeptides or proteins).
• amino acids such as arginine, asparagine, glutamine or histidine, carry reactive amino groups, which can react with the dihydroxy aromatic.
• the reactive amino groups are particularly suitable for the crosslinking reaction between peptide and oligolactone.
• hydroxyl groups or mercapto groups in the peptide may also contribute to the crosslinking reaction.
• the peptide therefore preferably comprises at least one amino acid with a hydroxyl group, ie in particular serine, threonine or tyrosine or a mercapto group, for example cysteine.
• Hydroxylysine or polyphenolic amino acid building blocks, as present in the MAPs can also be present in the peptides used according to the invention.
• An advantage of the present Invention is, however, that the presence of these specific amino acid building blocks and thus the use of MAPs is not necessary.
• the peptides used can therefore be prepared, for example, without further recombinant.
• the crosslinking produced by the reaction of the bridging molecules with the peptide depends essentially on how high the proportion of the available reactive groups in the peptide is. Good adhesive properties can be achieved from at least 10% of the amino acid content of the peptide carrying a reactive amino group (e.g., diamino acids such as lysine). If the polymer is not a peptide, the proportion of polymer structure elements which carry an amino group is at least about 10%. However, the proportion of these amino acids or structural elements is preferably higher, at least 20%, at least 30%, at least 40 or at least 50% or even at least 80 to 100%. Naturally occurring peptides and proteins, e.g. Albumin or casein may be used, particularly suitable are e.g. MAPs.
• Naturally occurring peptides and proteins e.g. Albumin or casein may be used, particularly suitable are e.g. MAPs.
• amino acids of the peptide are lysine.
• about 50% of the amino acids of the peptide may be tyrosine.
• lysine and tyrosine may be arranged as a repeating dipeptide moiety.
• a different sequence or the inclusion of other amino acids in particular arginine, asparagine, glutamine or histidine (instead of lysine or in addition), serine or threonine (instead of tyrosine or in addition), of cysteine or other amino acids is possible.
• a length of the peptide of about 10-20 amino acids is particularly preferred.
• the substituted polyhydroxyaromatic serves as an organic bridging molecule for cross-linking the peptide.
• proteins are used as substrates of the enzymes.
• bridge molecules should be used that are not of proteinogenic origin. It has surprisingly been found that substituted polyhydroxyaromatics have particularly favorable polymerization properties compared to unsubstituted polyhydroxyaromatics, in particular rapid polymerisation and good bond strength are achieved. A suitable substitution of the aromatics can even lead to monohydroxyaromatics being suitable for crosslinking as bridging molecule.
• Substituted means in the context of this invention that residues are bound to one molecule, in this case at the aromatic next to the two hydroxyl groups still 1, 2, 3 or 4 further radicals, one of these independently one or more radicals H, CH 3 , COH, COCH 3 , CONH 2 , CON-alkyl, CON-alkyl-OH, COOH, COO-alkyl, alkyl or may be a substituted aromatic, but not all radicals are H.
• the aromatic may also be part of a bicyclic or polycyclic aromatic ring system. Preference is given to a substituted dihydroxyaromatic 2 or 3 of the radicals H.
• substituted dihydroxyaromatics there are e.g. 2, 5-dihydroxybenzamide, preferably 2, 5-dihydroxy-N- (2-hydroxyethyl) benzamide, into consideration. It has been found that substituted dihydroxyaromatics are much better suited for the combination according to the invention than unsubstituted ones.
• Alkyl denotes branched or unbranched aliphatic hydrocarbon chains, preferably with 1-20, more preferably 1-6 Carbons, for example methyl, ethyl, propyl, butyl, isobutyl, n-pentyl, n-hexyl.
• the substituted polyhydroxyaromatic is a substrate of a polyphenol oxidase.
• the polyphenol oxidase serves as a catalyst for the reaction between peptide and polyhydroxyaromatic.
• the polyphenol oxidase is a lignolytic polyphenol oxidase, in particular laccase (EC 1.10.3.2).
• laccase EC 1.10.3.2
• lignolytic polyphenol oxidases such as laccase have a particularly broad substrate spectrum.
• Laccases are known in the art. They can come from plants or fungi or derived from natural enzymes.
• the laccases used in the invention may be recombinantly produced or purified. In this case, a particular purity of the laccase is generally not required. It is also possible to use supernatants of lignolytic fungi. However, for medical applications, substantial separation of microbiological substances such as lipopolysaccharides is desirable.
• laccases from the species Aspergillus, Neurospore, Podospora, Botrytis, Collybia, Fomes, Lentinus, Pleurotus, Pycnoporus, Pyricularia, Trametes, Rhizoctonla, Coprinus, Psatyrella, Mycelophtera, Schtalidium, Polyporus, Phlebia or Coriolus.
• the production of laccases is e.g. in EP 0 947 142.
• the peptide is an oligopeptide with a length of about 10-20 amino acids, preferably with a proportion of about 50% diamino acids such as lysine, the bridging molecule is 2,5-dihydroxybenzamide, preferably 2, 5 Dihydroxy-N- (2-hydroxyethyl) -benzamide, and the catalyst is laccase.
• crosslinking reaction proceeds in the combination according to the invention corresponding to that in Reaction Scheme 1 by way of example illustrated scheme.
• Formula 1 or 2 schematically illustrates a product obtained in which peptides / polymers are linked by the substituted dihydroxyaromatic.
• R 1, R 2 , R 3 and / or R 4 is independently H, OH, CH 3 , COH, COCH 3 , CONH 2 , C 1-4 alkyl, C 1-4 alkyl, COOH, COO alkyl, alkyl or substituted aromatic , in particular H, CH 3 , alkyl, or substituted aromatic. At least one of these radicals is not H. Preferably, three of these radicals are H and the fourth radical is CON-alkyl-OH, in particular CON-C 2 H 4 -OH.
• R6-R5 (-NH 2) -R 7 represents the amino-carrying peptide or polymer, wherein R6 or R7 may be H or both represent residues amino acid units or polymer units.
• R5 (-NH 2 / - NH-) is a diamino acid or a terminal amino acid of the peptide / amino group of the polymer with a free amino group or their reaction product.
• R 1 and / or R 2 independently of one another are H, OH, CH 3 , COH, COCH 3 , CONH 2 , CON-alkyl, CON-alkyl-OH, COOH, COO-alkyl, alkyl, substituted aromatic, in particular H, CH 3 , alkyl, or substituted aromatic, wherein not both radicals are H.
• R4-R3-R5 represents the amino group-bearing peptide / polymer, where R4 or R5 may be H or both residues represent amino acid units / polymer units.
• R3 (-NH-) is the reaction product of a diamino acid or a terminal amino acid of the peptide / amino group of the polymer having a free amino group.
• the individual components of the combination are present separately from one another before use, in particular components a and b are present separately from one another.
• a premix of components preparing the use may also be used, for example, the peptide / polymer and catalyst may already be mixed.
• the bridging molecule and peptide / polymer are mixed only when the reaction between the two is desired.
• a mixture comprising both peptide / polymer and bridging molecule may still be storable to some extent.
• one or more components, taken together or separately, may be included in one or more solvents.
• the solvent is non-toxic and biocompatible.
• the solvent is a phosphate buffer such as calcium phosphate or sodium phosphate buffer or PBS, but it may also be an organic solvent such as DMSO or a mixture of an aqueous and an organic solvent.
• the combination or its components can only be included in the solvent prior to use.
• the combination is already prepared for use in a two-component syringe, preferably already with an attached mixing extruder, wherein one component comprises the peptide and optionally the catalyst and the other component comprises the bridging molecule. This allows a particularly accurate dosage and easy handling.
• a double-chamber syringe of the type Mixpac (Mixpac Systems AG, Rotnch, Switzerland) is preferred.
• the components of the combination are already present in a quantitative ratio which is appropriate for their use and avoids inconvenient dosing of the ingredients.
• the proportion of peptides in the combination can be e.g. about 20-99% (the percentages are based on wt .-%, based on the total mass without solvent), preferably about 40-90%, amount.
• the proportion of bridging molecules may be e.g. about 1-80%, preferably about 5-70%.
• the proportion of polyphenol oxidase is about 0.001-5%, preferably about 0.01-1% or about 0.05-0.5%.
• the proportions can be adjusted in relation to the desired application.
• a proportionately larger amount of bridge molecule leads to a higher degree of crosslinking.
• reactive amino, mercapto and / or hydroxyl groups of substrates to be bonded are increasingly included in the crosslinking, if a lower proportion of peptide is present.
• the amount of enzyme affects the rate of the reaction, with, depending on the application, a rapid achievement of the gel point or complete curing or a longer processability the combination may be desired. An optimization is possible through experiments.
• the combination may contain other additives and adjuvants, e.g. Fillers such as collagen, albumin, hyaluronic acid or the like.
• Fillers such as collagen, albumin, hyaluronic acid or the like.
• the total content of peptide and bridging molecule is about 25-100%, especially about 50-90%.
• wound healing promoting substances e.g. Growth factors, or antibiotic active substances may be included.
• amino acid-containing antimicrobial agents modified by the action of the components of the invention and fixed to surfaces, resulting in an antimicrobial finish. Such modifications are e.g. for the treatment of infected wounds of interest.
• the combination according to the invention is in particular an adhesive, i.e. it is suitable for bonding substrates, e.g. of tissue.
• the combination according to the invention is preferably present as a medical product.
• the medical device is suitable for bonding tissue but may also be used to adhere other substrates, e.g. Implants are used. Classification as a pharmaceutical preparation is also possible, depending on national law. These terms are interchangeable for the purposes of describing the invention.
• the present invention provides a use of the combination for making a medical device for bonding tissue.
• a sterilization of the medical device or its individual components can be advantageously achieved without structural change. Possible, for example, is a sterile filtration of solutions. However, sterilization by means of gamma radiation is preferred since it can already be packed and therefore no aseptic filling is necessary. The loss of activity of laccase in gamma sterilization, which is up to 50% can be easily compensated by a corresponding higher initial concentration.
• the medical device can be used to glue hard or soft tissue.
• connective tissue, skin, tendon, organ, blood vessel and / or nerve (soft tissue) and / or tooth and / or bone (hard tissue) may be adhered.
• various tissues can be glued together, e.g. Tendons and bones.
• a particularly preferred application is in the bonding of wound edges or in the adhesion of ruptures of various organs, such as liver, kidney or spleen.
• the care of surgically placed wounds e.g. after the removal of tumors, is possible with the medical device according to the invention.
• the advantages of the combination according to the invention enable its use in particularly difficult cases, in particular also in the treatment of chronic wounds. With such wounds, after the necessary surgical treatment, a strong tension is created, which hampers wound healing.
• the high strength of the wound closure according to the invention also enables a firm seal in these cases and prevents wound infections.
• compressive bandages may be applied to the wounds so closed to prevent edema formation.
• the combination can be used for fixation of implants, eg vascular prostheses, biodegradable implants, catheters, stents and other materials.
• pharmaceutical active substances are incorporated into the adhesive system provided for fixing implants so as to prevent infections, prevent unwanted cell growth and accelerate healing. Absorption and healing can be coordinated with each other with the help of the stored pharmaceutical agents.
• An advantageous application of this combination is the flexible, over a certain period resorbable sealing of anastomoses and patches on vessels or on hollow organs.
• the combination is also suitable for the fixation of drug-delivery devices and porous carrier structures or membranes for potential use in regenerative medicine (tissue engineering).
• the developed medical products can also be used excellently for bonding hard tissue, for example for bone bonding.
• hard tissue for example for bone bonding.
• the acceptance of the implanted material by the bone cells is of fundamental importance.
• the fast colonization with osteoblasts and their subsequent maturation promotes new bone formation and shortens the healing process.
• Implants are therefore preferably made with microstructured surfaces.
• the tissue adhesive according to the invention preserves this structure and supports the formation of new bone.
• collagen which is indispensable for bone healing, the amino acids lysine and proline are necessary. Lysine is included at the splice site on a large scale.
• the extracellular matrix formed is recognized by integrins such as ⁇ l ⁇ 1 or ⁇ 3 ⁇ 1 from the RGD sequence (Arg-Gly-Asp). This initiates a signal cascade that triggers changes in the cytoskeleton in the cell and switches between the proliferation stage and the stage of differentiation.
• the protein matrix formed also serves to store calcium salts.
• the medical device according to the invention therefore preferably serves for fixing debris fractures, e.g. in the facial skull area or the extremities.
• the combination according to the invention is also particularly suitable for the production of medical devices for fixing muscles, ligaments and tendons to bones.
• Another application is in the field of dentistry, e.g. in the bonding of inlays and crowns or in bodywork.
• the viscosity and flowability of the combination used can be adjusted depending on, for example, the length of the wound to be fixed and the wound gap depth or the substrates to be bonded. In addition to the length of the peptides / polymers used, the amount of solvent also has an influence on these parameters. Also, additives such as thixotropic agents can be used to adjust viscosity and flowability. Typically, for bonding hard tissue, a higher viscosity medical device is used than for soft tissue bonding.
• An advantage of the combination according to the invention is that the compound formed by reaction of the components is biodegradable, in particular when using peptide as polymer.
• the crosslinked combination is resorbed in the body within about one year (biodegraded), preferably in a period of about 28 days to about 6 months.
• the mechanism of absorption may be e.g. be hydrolytic or enzymatic.
• the peptides can be cleaved and individual fragments removed and excreted. Alternatively, fragments or amino acids can also be incorporated into the regenerating tissue.
• the invention provides a process for the preparation of linked polymers in which the components of the combination according to the invention are brought into contact with one another.
• the resulting products are highly molecularly crosslinked and may, for example, depending on the starting components, in this case preferably peptides / proteins which are suitable for consumption, be used for the production of foodstuffs.
• the tissue-adhering combination of the present invention is used to make a medical device for treating bleeding wounds used in combination with a hemostatic agent.
• a hemostatic agent is provided which can be used together in bleeding wounds. It has It has been pointed out that a novel hemostatic agent is particularly suitable in this context.
• the present invention therefore provides a combination for the treatment of bleeding wounds comprising
• a hemostatic agent comprising a zeolite fine granules having a zeolite content of at least 70% and an average pore size of 0.3 to 0.5 nm, obtainable by dehydration at temperatures ⁇ 200 ° C.
• a combination for bonding tissue comprising a) a polymer, in particular a peptide which comprises at least two amino groups, and b) a substituted polyhydroxyaromatic, in particular dihydroxyaromatic, which is not a peptide component, and c) a polyphenol oxidase, in particular a laccase.
• the combination can be the combination according to the invention in all the embodiments described in detail above.
• the zeolite fine granules are preferably a natural zeolite granules containing clinoptilolite, chabazite and / or mordenite.
• the zeolite is fractionated after drying and pre-crushing. Hemostasis is achieved by natural zeolite granules having a micropore space with an average pore size of 0.3 to 0.5 nm. It has been found that in the microporous space of these minerals, certain factors involved in blood coagulation are selectively enriched. This leads to an activation of physiological blood coagulation, which leads to a hemostyptics significantly superior hemostasis. It is of particular advantage that no toxic substances are released to the wound. Selected Bacholithe can be processed according to the invention so that they can now be provided cost-effective direct way as hemostatic agents and means for wound care. The fraction is used with grain band 0.8 - 0.2 mm. Regarding purity, the requirements are US Pharmacop. or Europ. Pharmacorp. to fulfill. A gentle dehydration is preferably carried out at temperatures below 200 0 C by treatment at temperature intervals.
• the dehydration is preferably only partial so that upon ingestion of liquid, e.g. Blood, no strong, tissue-damaging exothermic reaction occurs.
• liquid e.g. Blood
• the Zeolithfeingranulat ⁇ 200 0 C is therefore by grinding, in particular from natural zeolite with a content of clinoptilolite, chabazite and / or mordenite and fractionation and gentle partial dehydration at temperatures pretreated.
• a granulated mineral granulate with strong electrostatic fields in the crystal lattice is obtained which is able to stop even heavy bleeding within 1 min.
• the product according to the invention on the market products based on microporous synthetic lithium silicates (EP 1176991 Al, WO 00/69480, US 6833486 Bl), z.
• CERDAC is superior by a 5-10-fold higher capillary force effect.
• no exothermic reaction occurs, which can cause tissue damage by combustion.
• the high capillary force is also of particular importance for use in wound care, in particular for the treatment of wounds that are difficult to heal and have already become chronic. Particularly indicated is the use in wounds with an exudation, ie an escape of blood components on an inflamed wound. Often an exudation is an indication of an infection on an injury.
• the microporous structure obtained according to the invention enables the uptake of wound exudate, At the same time, ingesting the excess exudate creates a humid atmosphere over the wound, which is important for wound healing.
• the hemostatic agent may further comprise biomass from aquatic organisms, e.g. Algae, included.
• biomass from aquatic organisms e.g. Algae
• the minerals with a pore width of 0.3 to 0.5 nm in the ratio 1:10 to 10: 1 can advantageously be mixed with the biomass from aquatic organisms.
• microalgae with antibacterial activity, lyophilizes and grinds their biomass and mixes them with the minerals (zeolites), so that germ colonization of treated wounds is prevented.
• microalgae are selected with a lipid content of> 20% and many unsaturated fatty acids, since these have a high antibacterial activity and / or contain poly-N-acetyl-glucosamine.
• microalgae promote cell proliferation and have an anti-inflammatory and immunostimulating effect.
• freeze-dried biomasses of the green alga Chlamydomonas and / or the microalgae Spirulina and / or Anabaena are used.
• the fine granules of natural zeolite and microalgae biomass can also be used for the production of medical products in order to treat severely infectious, weeping wounds, which can then be closed with tissue glue, if necessary.
• the hemostatic agents contained in the combination according to the invention may additionally comprise collagen, on one Collagen matrix can be applied or used to fix collagen.
• the components of the invention as well as further additives can be applied in different layers to a collagen matrix.
• the biomass of the aquatic organisms used contains lipids, which support the phagocytosis and thus lead to a complete absorption of the collagen parallel to the wound healing.
• the biomass may e.g. dissolved and / or suspended in collagen hydrolysates and applied to a collagen matrix by lyophilization.
• the mineral for hemostasis in a sheath e.g. based on cellulose or on a textile basis or based on a collagen fleece. After hemostasis, the mineral can be completely removed in this case. By activating the fibrin system, the wound edges are glued together. However, the tear strength of the wound closure is lower than with conventional fibrin adhesives.
• the tissue adhesive according to the invention described above is provided, with which the wound can be adhered in a second step according to the invention.
• the inventive products provide a particular advantage in their overall effect, which is that now hemostyptics and tissue adhesives can be provided, which complement each other in various applications advantageous.
• the present invention teaches the use of the described combination to produce a medical device.
• the medical device is suitable for supplying bleeding or weeping wounds. This supply is done by in particular by haemostasis and subsequent adhesion of the wound edges.
• the mineral components thereby create a blood-free surgical field.
• the wound adhesive therefore, the wound is clearly visible. This allows the exact local assignment of the wound edges.
• the fibrin system By activating the fibrin system, the wound edges are glued together. There is no danger that the components of the adhesive required for the gluing process, for example the polyphenol oxidases, will be diluted or washed away by the bleeding. It is also ruled out that components of the adhesive system are washed away by a strong bloodstream and lead to adhesive reactions in other tissue regions. The risk of thrombosis associated with conventional fibrin glue is drastically reduced.
• the adhesives of the invention are also much easier to handle. Special storage requirements such as frozen fibrin glue do not exist.
• the laccase may e.g. be used in dissolved form, then storage at refrigerator temperature is sufficient. It is also possible to deliver the laccase in powder form and to solve it before a foreseeable application.
• the adhesives according to the invention are an alternative both for electrocoagulation and for wound closure with needle and thread
• Example 1 Preparation of the natural zeolite fine granules
• Example 2 Testing of the microporous volume and the particle size distribution
• the products prepared according to Example 1 have 25-50% micropore space (based on the total volume) and average nanopore diameters in the range 0.3-0.5 nm.
• Nanopore size and fine granulate structure allow maximum blood adsorption and rapid penetration of the adsorber fine granules.
• the capillary force effect is 5 to 10 times higher than in the comparatively investigated commercial preparation CERDAC
• Example 3 Testing the fine granules for suitability for haemostasis
• Rats were anesthetized for this purpose. After opening the abdominal cavity was with the Scalpel made a cut about 1 cm long. The bleeding was filmed. The time to hemostasis was measured.
• hemostasis is achieved within 1 min (control without treatment up to 8 min). It forms a scab-like encrustation that protects the wound.
• Example 4 In vitro examination of the fine granules for suitability for haemostasis
• Acetate tissue as used for the preparation of wound dressings, is dripped with 2 mg of extract equivalent amount of extract solution and allowed to dry.
• no pathogens can be used Wound infections detect dreaded methicillin-resistant staphylococcal strains (MRSA). In other staphylococci, germ growth was reduced to 10 to 20% of the starting bacterial count.
• MRSA methicillin-resistant staphylococcal strains
• Example 6 Production of fine granules from natural zeolites and the biomass of microalgae
• the mineral granules produced according to Example 1 microalgae biomasses with a lipid content> 20% in the ratio 1:10 to 10: 1 are added. This prevents the germ colonization of the wounds.
• Example 7 Testing the fine granules for suitability for haemostasis and wound closure
• Rats were anesthetized for this purpose. After opening the abdomen, the scalpel was used to make a cut of about 1 cm. The bleeding was filmed. The time to hemostasis was measured. The wound closure was assessed visually.
• hemostasis is achieved within 1 min (control up to 8 min). It forms a scab-like encrustation that protects the wound. b) In further experiments, the encrustation was removed after 1 min with the scalpel.
• wound closure is better for fine granules based on mineral / algae biomass than for the preparations made exclusively on a mineral basis.
• Example 8 Preparation of the adhesive prepolymers and performing the adhesive reaction for soft tissue and hard tissue adhesion
• bridging molecule 2 2, 5-dihydroxy-N- (2-hydroxyethyl) benzamide
• laccase 5 the enzyme laccase
• Adhesion occurs through the enzymatic reaction of the laccase with the bridging molecule linked to the oligopeptide.
• the tissue parts to be examined had to be fixed beforehand. Porcine soft tissue was fixed to polymethylmethacrylate (PMMA) specimen holders with cyanoacrylate adhesive, or bone plates of bovine bone were clamped in mechanical clamps. Subsequently, the adhesive mixture to be examined (see Example 6) was applied to the fabric.
• PMMA polymethylmethacrylate
• the strength of the adhesive joint under tensile stress on the Zwick BZ2.5 / TN1S testing machine (test speed: 10 mm / min) was determined after 10 minutes.
• the mechanical testing of the adhesive joint after hard tissue bonding was carried out under tensile shear stress at a test speed of 5 mm / min. Result: Twice as strong as the comparatively tested commercial fibrin adhesives.
• Example 10 Diffusion-inhibition test with the inventively doped collagen fleece in comparison to a nonwoven doped with a conventional antibiotic.
• the agar diffusion test was carried out according to Burkhardt (Burkhardt, F. (ed.) Mikrobiologische Diagnostik, Georg-Thieme Verlag Stuttgart, New York 1992, p 724).
• Burkhardt Backhardt, F. (ed.) Mikrobiologische Diagnostik, Georg-Thieme Verlag Stuttgart, New York 1992, p 724).
• Mueller-Hinton II agar was used in Stacker petri dishes (Becton Dickinson Microbiology Systems, Cockeysville, USA).
• S. aureus strain ATCC 6538 was selected. The seeding of this test strain was chosen so that after 1620 h of incubation closely spaced but not confluent single colonies develop.
• the doped collagen fleeces are placed on the agar surface. After 18 + 2 h incubation at 36 0 C, the inhibition zones are measured
• Agar plate (Müller-Hinton II Agar ready-made plates from Becton Dickinson) distributed.
• Polymer biomaterials (1 cm 2 each) are named after Contrib. Plasma Phys. 41, 2001, 562-572 functionalized in ammonia plasma. Thereafter, the reaction was carried out with 2,5-dihydroxy-N- (2-hydroxyethyl) benzamide (12.5 mM) under the influence of laccase (0.32 U (156 nmol ml "1 min '1 ) Methanol was 10% (v / v).
• Example 12 Use of the combination according to the invention for the treatment of bleeding wounds.
• Rats are anesthetized as in Example 7, the abdomen is opened and a section is taken in the liver. The bleeding is quenched as in Example 7c. The forming crust is removed. Then the cut is glued with adhesive.
• the oligopeptides described in Example 6 are taken up in PBS (Phosphate Buffered Saline, 2.7 M NaCl, 54 mM KCl, 87 mM Na 2 HPO 4 , 30 mM KH 2 PO 4 , pH 7.4) and laccase added (Component 1).
• PBS Phosphate Buffered Saline, 2.7 M NaCl, 54 mM KCl, 87 mM Na 2 HPO 4 , 30 mM KH 2 PO 4 , pH 7.4
• laccase added Component 1
• 2, 5-Dihydroxy-N- (2-hydroxyethyl) benzamide is dissolved in PBS (component 2).
• oligopeptide / 2, 5-dihydroxy-N- (2-hydroxyethyl) benzamide / laccase are tested, eg 1/1 / 0.1; 1/2 / 0.5; 1/10/1)
• the amount of solvent is minimally selected in order to achieve the most concentrated possible solution of the components.
• the components are placed in a two-component syringe with mixpac mixing extruder (Mixpac Systems, Rotnch, Switzerland) and introduced into the wound gap.
• Collagen film (DOT GmbH) was fixed on a flat surface.
• the collagen film was divided into 8 segments (4 each in two rows) and the adhesive was mixed together directly on the film (taking advantage of the surface tension).
• the glue was dissolved in phosphate citrate buffer. Different concentrations of bridging molecule (laccase substrate) were tested.
• the adhesive protein had an MW of about 4000D). Subsequently, small collagen pieces were applied, lightly pressed and allowed to dry.
• Adhesive solution (buffer, laccase, laccase substrate) was applied to the
• the collagen has been permanently bonded to the ground.
• the cell adhesion was carried out on 60 mm TCPS cell culture dish, which was coated according to Example 15 with collagen. After staining the cells with crystal violet, the evaluation was carried out with the
• the FL cells become more bound.
• EXAMPLE 16 Coating for Short Percutaneous Catheters A catheter tube made of polyurethane, for example polyetherurethane, provided for a short-term catheter is plasma-treated in a microwave plasma at a pressure of 0.1 mbar in ammonia at a power of 500 W for 10 seconds. The resulting in the result 2% of amino groups on the surface are reacted with 10 mmol of 2,5-dihydroxy-N- (2-hydroxyethyl) benzamide under the influence of laccase (0.32 U (156 nmol ml "1 min " 1 ) Treated catheter tubing is incorporated as a percutaneous section in catheter systems, causing a reduction in the risk of infection.
• polyurethane for example polyetherurethane
• a thin flexible polydimethylsiloxane cannula designed for a long-standing fixed drug delivery system (for example for the local treatment of tumors) is placed in an argon microwave plasma with an admixture of 1% ethylene diamine at a pressure of 0.5 mbar at a power of 500 W plasma treated for one minute. This forms a crosslinked film of ethylene diamine, which covers the surface and contains amino groups.
• the inserted drug delivery system is permanently fixed at the destination for a long time.
• Example 18 Bindable titer plate coating
• Titer plates of optically transparent, inert plastics for example of polystyrene, polycarbonate, cyclic olefin copolymer or polypropylene are plasma-treated in a microwave plasma at a pressure of 0.1 mbar in ammonia at a power of 500 W for 10 seconds.
• a bond-chemical structure is carried out on amino groups.
• This is followed by the addition of the polyphenols and the lacase and the functional molecules.
• the resulting stereochemistry-unaffected surface is used to attach functional molecules.

Landscapes

• Health & Medical Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• Medicinal Chemistry (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Polymers & Plastics (AREA)
• Animal Behavior & Ethology (AREA)
• Veterinary Medicine (AREA)
• Public Health (AREA)
• General Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Pharmacology & Pharmacy (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Engineering & Computer Science (AREA)
• Bioinformatics & Cheminformatics (AREA)
• General Chemical & Material Sciences (AREA)
• Surgery (AREA)
• Dermatology (AREA)
• Epidemiology (AREA)
• Vascular Medicine (AREA)
• Heart & Thoracic Surgery (AREA)
• Diabetes (AREA)
• Hematology (AREA)
• Cardiology (AREA)
• Communicable Diseases (AREA)
• Oncology (AREA)
• Materials For Medical Uses (AREA)
• Adhesives Or Adhesive Processes (AREA)
• Peptides Or Proteins (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Preparation Of Compounds By Using Micro-Organisms (AREA)
• Medicinal Preparation (AREA)

Applications Claiming Priority (2)

Publications (1)

Family

ID=38261578

Family Applications (5)

Family Applications Before (2)

Family Applications After (2)

Country Status (10)

Families Citing this family (30)

Family Cites Families (45)

• 2006
  • 2006-02-09 DE DE102006006904A patent/DE102006006904A1/de not_active Withdrawn
• 2007
  • 2007-02-02 ES ES15202504T patent/ES2771824T3/es active Active
  • 2007-02-02 EP EP15202504.5A patent/EP3050579B1/fr active Active
  • 2007-02-02 ES ES07702422.2T patent/ES2566054T3/es active Active
  • 2007-02-02 WO PCT/DE2007/000192 patent/WO2007090373A2/fr active Application Filing
  • 2007-02-02 EP EP07702422.2A patent/EP1984032B1/fr active Active
  • 2007-02-09 AT AT07704469T patent/ATE504317T1/de active
  • 2007-02-09 AU AU2007213923A patent/AU2007213923C1/en active Active
  • 2007-02-09 WO PCT/EP2007/001131 patent/WO2007090673A2/fr active Application Filing
  • 2007-02-09 US US12/223,899 patent/US20090280179A1/en not_active Abandoned
  • 2007-02-09 EP EP07721909A patent/EP1987111A2/fr not_active Withdrawn
  • 2007-02-09 DE DE502007006879T patent/DE502007006879D1/de active Active
  • 2007-02-09 EP EP07711497.3A patent/EP1981553B1/fr active Active
  • 2007-02-09 WO PCT/EP2007/051251 patent/WO2007090880A2/fr active Application Filing
  • 2007-02-09 CA CA2637794A patent/CA2637794C/fr active Active
  • 2007-02-09 CN CN2007800051590A patent/CN101384284B/zh active Active
  • 2007-02-09 US US12/223,772 patent/US7923003B2/en active Active
  • 2007-02-09 JP JP2008553771A patent/JP2009525799A/ja not_active Withdrawn
  • 2007-02-09 CN CNA2007800047684A patent/CN101378789A/zh active Pending
  • 2007-02-09 JP JP2008553686A patent/JP2009525795A/ja active Pending
  • 2007-02-09 AU AU2007213663A patent/AU2007213663A1/en not_active Abandoned
  • 2007-02-09 WO PCT/DE2007/000241 patent/WO2007090384A2/fr active Application Filing
  • 2007-02-09 CA CA2640771A patent/CA2640771C/fr not_active Expired - Fee Related
  • 2007-02-09 EP EP07704469A patent/EP1981552B1/fr not_active Not-in-force
• 2011
  • 2011-03-01 US US13/037,850 patent/US8440209B2/en not_active Expired - Fee Related

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events