Classifications

• • 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
  • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
  • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
  • A61L27/54—Biologically active materials, e.g. therapeutic substances
• • 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
  • A61L29/00—Materials for catheters, medical tubing, cannulae, or endoscopes or for coating catheters
  • A61L29/14—Materials characterised by their function or physical properties, e.g. lubricating compositions
  • A61L29/16—Biologically active materials, e.g. therapeutic substances
• • 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
  • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
  • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
  • A61L31/16—Biologically active materials, e.g. therapeutic substances
• • G—PHYSICS
  • G02—OPTICS
  • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
  • G02B1/00—Optical elements characterised by the material of which they are made; Optical coatings for optical elements
  • G02B1/04—Optical elements characterised by the material of which they are made; Optical coatings for optical elements made of organic materials, e.g. plastics
  • G02B1/041—Lenses
  • G02B1/043—Contact lenses
• • 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
  • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
  • A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
  • A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
• • 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
  • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
  • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
  • A61L2300/404—Biocides, antimicrobial agents, antiseptic agents

Definitions

• This invention relates to coated devices.
• the invention provides biomedical devices on the surfaces of which antimicrobial coatings of a cationic peptide, a cationic protein, or both are formed.
• Devices for use in and on the human body are well known.
• the chemical composition of the surfaces of-such devices plays a pivotal role in dictating the overall efficacy of the devices. Additionally, it is known that providing such devices with an antimicrobial surface is advantageous.
• bactericidal and bacteriostatic coatings have been developed.
• cationic antibiotics such as polymyxin, vancomycin, and tetracycline have been used as coatings for contact lenses.
• metal chelating agents substituted and unsubstituted polyhydric phenols, aminophenols, alcohols,
• Chelating agent use fails to address the numbers of bacteria that adhere to the device.
• Some of the prior art coatings for example phenol derivatives and cresols, can produce ocular toxicity or allergic reactions.
• Quarternary ammonium compounds are problematic because of their irritancy.
• the present invention provides biomedical devices with an antimicrobial coating and processes for the production of the biomedical devices. It is an unexpected discovery of the invention that certain cationic peptides, cationic proteins, or both may be used to provide antimicrobial coatings for biomedical devices. In particular, it is one discovery of the invention that protamine, melittin, cecropin A, nisin, or combinations thereof, when used as surface coatings, reduce adherence of bacteria to a device's surface, reduce growth of bacteria adhered to a device, or both by greater than about 50 percent.
• the invention provides a biomedical device comprising, consisting essentially of, and consisting of at least one surface comprising, consisting essentially of, and consisting of a coating effective amount of one of protamine, melittin, cecropin A, nisin, or combinations thereof.
• a method for manufacturing biomedical devices comprising, consisting essentially of, and consisting of contacting at least one surface of a biomedical device with a coating effective amount of protamine, melittin, cecropin A, nisin, or combinations thereof is provided.
• biomedical device any device designed to be used while in or on either or both human tissue or fluid. Examples of such devices include, without limitation, stents, implants, catheters, and ophthalmic lenses.
• the biomedical device is an ophthalmic lens including, without limitation, contact or intraocular lenses. More preferably, the device is a contact lens, most preferably a soft contact lens.
• Protamine is isolatable from the sperm of a variety of animals including, without limitation, man. Melittin is isolatable from bee venom. Cecropin A and nisin are isolatable from Aedes aegypti and Lactoccucus lactis, respectively. All four are members of a broad group of cationic peptides and proteins which group includes, without limitation, defensins, magainins, and colicins. It is an unexpected discovery of this invention that only certain cationic peptides and proteins significantly reduce bacterial adherence, bacterial growth, or both on biomedical devices.
• Protamine, melittin, cecropin A, and nisin useful in the invention are all commercially available.
• these cationic peptides and proteins may be produced by known means.
• the purity of the cationic peptide used is at least about 75 %, preferably at least about 90 %.
• Protamine, melittin, cecropin A, nisin, or combinations thereof may be adsorbed to polymer surfaces of a biomedical device.
• the cationic peptides and proteins may be used on any surface, but most advantageously are used with negatively charged surfaces.
• the cationic peptides and proteins alternatively may be bound to the polymer surfaces.
• This may be either a direct reaction or, preferably, a reaction in which a coupling agent is used.
• a direct reaction may be accomplished by the use of a reagent of reaction that activates a group in the surface polymer or the cationic peptide making it reactive with a functional group on the peptide or polymer, respectively, without the incorporation of a coupling agent.
• one or more amine groups on the peptide may be reacted directly with isothiocyanate, acyl azide, N-hydroxysuccinimide ester, sulfonyl chloride, an aldehyde, glyoxal epoxide, carbonate, aryl halide, imido ester, or an anhydride group on the polymer.
• coupling agents may be used.
• Coupling agents useful for coupling the cationic peptide or protein to the device's surface include, without limitation, N, N'-carbonyldiimidazole, carbodiimides such as l-ethyl-3-(3- dimethylaminopropyl)carbodiimide (' ⁇ DC"), dicyclohexyl carbodiimide, 1- cylcohexyl-3-(2 ⁇ morpholinoethyl)carbodiimide, diisopropyl carbodiimide, or mixtures thereof.
• the carbod ⁇ mides also may be used with N-hydroxysuccinimide or N- hydroxysulfosuccinimide to form esters that can react with amines to form amides.
• Amino groups also may be coupled to the polymer by the formation of Schiff bases that can be reduced with agents such as sodium cyanoborohydride and the like to form hydrolytically stable amine links.
• Coupling agents useful for this purpose include, without limitation, N-hydroxysuccinimide esters, such as dithiobis(succinimidylpropionate), 3,3'-dithiobis(sulfosuccinimidylpropionate), disuccinimidyl suberate, bis(sulfosuccinimidyl) suberate, disuccinimidyl tartarate and the like, imidoesters, including, without limitation, dimethyl adipimate, difluorobenzene derivatives, including without limitation l,5-difluoro-2,4- dinitrobenzene, bromofunctional aldehydes, including without limitation gluteraldehyde, and bis epoxides, including without limitation 1,4-butanediol dig
• the device's surface does not contain suitable reactive groups
• suitable groups may be incorporated into the polymer by any conventional organic synthesis methods.
• the reactive groups may be introduced by the addition of polymerizable monomers containing reactive groups into the monomer mixture used to form the polymer.
• polymer surfaces onto which the cationic peptides and proteins may be adsorbed or bonded are surfaces formed from, without limitation, polymers and copolymers of styrene and substituted styrenes, ethylene, propylene, acrylates and methacrylates, N-vinyl lactams, acrylamides and methacrylamides, acrylonitrile, acrylic and methacrylic acids as well as polyurethanes, polyesters, polydimethylsiloxanes and mixtures thereof.
• Such polymers may include hydrogels and silicpne containing hydrogels.
• lightly crosslinked polymers and copolymers of 2-hydroxyethyimethacrylate (' ⁇ EMA”) are used.
• lightly crosslinked is meant that the polymer has a low enough crosslink density so that it is soft and elastic at room temperature.
• a lightly crosslinked polymer will have about 0.1 to about 1 crosslinking molecule per about 100 repeating monomer units.
• suitable lightly crosslinked HEMA polymers and copolymers include without limitation, etafilcon A and copolymers of glycerol methacrylate and HEMA.
• silicone hydrogels especially those of hydrophilic monomers, such as N,N-dimethylacrylamide, are used.
• the surface to be coated is contacted with the protamine, melittin, cecropin A, nisin or combinations thereof in any convenient manner.
• the device may be placed in a solution of protamine and solvent into which the medical device is placed.
• the device's surface may first be treated with a coupling agent and the surface then placed in a solution of the selected cationic peptide or protein.
• the peptide or protein may be reacted alone with the polymer surface.
• Suitable solvents for use in the invention are those that are capable of dissolving protamine, melittin, cecropin A, or nisin singly or in combination.
• the coating process is carried out in water, alcohol, or mixtures thereof.
• EDC is effective in aqueous solutions and, thus, is a preferred coupling agent.
• the coupling agents may be used alone or in combination with agents capable of stabilizing any reactive intermediate formed.
• EDC may be used with N-hydroxysuccinimide as a stabilizer.
• the pH is adjusted to about 6.5 to about 8.0, more preferably about JO to about 7.5.
• a coupling effective amount of a coupling agent is used which amount is an amount sufficient to couple the peptide or protein to the device surface.
• the precise amount of coupling agent used will depend on the surface's chemistry as well as the agent selected. Generally, about 0.01 to about 10 weight percent, preferably about 0.01 to about 5.0, more preferably about 0.01 to about 1 weight percent of the coupling agent is used based on the weight of the coating solution.
• coating solution is meant the peptide or protein with one or more of the solvent, coupling agent, buffer, and the like.
• the amount of coating solution used per lens will be about 1 ml to about 10 ml, preferably about 1 ml to about 5 ml, more preferably about 1 ml to about 2 ml per lens.
• a coating effective amount of protamine, melittin, cecropin A, nisin, or combinations thereof is used meaning an amount that when contacted with the surface is sufficient to coat the surface so as to impart the desired antimicrobial properties to the surface.
• antimicrobial properties is meant either or both the ability to significantly reduce, meaning by greater than about 50 percent, either or both the amount of bacteria adhering to the surface and the growth of bacteria adhered to the surface.
• the amount contacted with the lens is about 1 ⁇ g to about 10 mg, preferably about 10 ⁇ g to about 1 mg per lens.
• the amount of coating resulting per contact lens is about 50 to about 1000 ⁇ g.
• the amount of protamine used preferably is about 500 ⁇ g/ml or less.
• the contact time used will be a length of time sufficient to coat the surface to the extent desired. Preferably, contact time is about 60 seconds to about 24 hours.
• the surface may be washed with water or buffered saline solution to remove unreacted protamine, melittin, colicin and solvent.
• the polymer for producing the surface to be coated by the method of the invention may contain other monomers and additives.
• ultra-violet absorbing monomers, reactive tints, processing aids, and the like may be used.
• **By "CLARE” is meant contact lens induced red eye.
• CPU contact lens-induced peripheral ulcers
• P. aeruginosa and S. aureus are the most common bacteria causing eye inflammation or infections for contact lens wearers.
• Other strains were used to validate results or assess the effectiveness of the compounds over a range of bacteria.
• Example 1 To assess the effect of the cationic proteins/peptides in solution against rapid growing bacterial cells, bacteria were cultured in Trypticase soy broth (' SB") overnight at 35°C. Aliquots (20 ⁇ l) of this cell suspension were then added to fresh TSB (10ml). Different concentrations of the cationic proteins/peptides were added to the fresh broth and incubated for up to 48h at 35°C. Samples were taken at different time points and the optical density at 660nm measured as a measure of changes in bacterial numbers was measured.
• TSB Trypticase soy broth
• cationic proteins/peptides were grown as previously in TSB. The cells were then harvested by centrifugation and washed in phosphate buffered saline (PBS; NaCl 8g/l; KC1 0.2g/l; Na 2 HPO 4 1.15g/l; KH 2 PO 4 0.2g/l). The cells were then re-suspended to OD 0.1 (unless otherwise stated) at 660nm in PBS, different concentrations of cationic proteins/peptides were added and incubated for up to 48h at 35°C. Samples were taken a different points and numbers of bacteria analyzed using the Miles and
• Misra technique i.e. numbers that are viable after plating dilutions onto nutrient agar plates.
• a "- " sign indicates no reduction in bacterial growth
• a "+” sign indicates a 1 to 50 % reduction in growth
• a "++” sign indicates a 51 to 89 % reduction in growth
• a "-H-+” sign indicates a 90 to 98 % reduction in growth
• a "++++” sign indicates a greater than 98 % reduction in growth.
• Example 2 For conducting total counts, etafilcon A lenses were removed from the manufacturers vials, washed three times in 1ml PBS and then coated with various concentrations of cationic proteins/peptides overnight at 37°C either individually or in combination. After incubation, the lenses were washed three times in PBS and 0.5ml of lxl 0 8 bacterial cells/ml was added to the lenses. After incubation at ambient temperature for lOmin, the lenses were washed three times in PBS to remove non- adherent or loosely adherent bacteria and stained with crystal violet for 1 min. The number ofcells per lens was examined under the microscope. Five grids (0.005625 mm 2 ) per lens were counted and triplicate lenses for each treatment were assayed.
• etafilcon A lenses were removed from the manufacturers vials, washed three times in 1ml PBS and then coated with various concentrations of cationic proteins/peptides overnight at 37°C (either individually or in combination). After incubation, the lenses were washed three times in PBS and 0.5ml of lxlO 8 bacterial cells/ml was added to the lenses. After incubation at ambient temperature for lOmin, the lenses were washed three times in PBS to remove non- adherent or loosely adherent bacteria.
• Lenses were then homogenized using 1 ml PBS and a small magnetic stirring bar (octagonal cross-section, 0.5" X 0.125”) and stirred at maximum speed for one hour which was sufficient for lens disintegration. Serial dilutions were then made according to the technique of Miles and Misra and aliquots (20 ⁇ L) plated out on nutrient agar. After incubation overnight at 37°C, viable bacteria were determined and results expressed as colony forming units/mm 2 after calculation of the surface area of the lens (approximately 310mm 2 ).
• the lenses were incubated in concentrations of cationic proteins/peptides that were either effective in solution or the highest concentration available if there was no effect in solution. After rinsing, bacteria were added and numbers ofcells analyzed as the total cells per r m of the lens or the number of viable cells per mm of the lens. The results are shown on Table 4.

Landscapes

• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Medicinal Chemistry (AREA)
• Chemical & Material Sciences (AREA)
• Public Health (AREA)
• Biomedical Technology (AREA)
• Molecular Biology (AREA)
• Epidemiology (AREA)
• Animal Behavior & Ethology (AREA)
• General Health & Medical Sciences (AREA)
• Engineering & Computer Science (AREA)
• Veterinary Medicine (AREA)
• Physics & Mathematics (AREA)
• General Physics & Mathematics (AREA)
• Optics & Photonics (AREA)
• Heart & Thoracic Surgery (AREA)
• Surgery (AREA)
• Vascular Medicine (AREA)
• Dermatology (AREA)
• Oral & Maxillofacial Surgery (AREA)
• Transplantation (AREA)
• Eyeglasses (AREA)
• Materials For Medical Uses (AREA)

Applications Claiming Priority (1)

Publications (1)

Family

ID=21742336

Family Applications (1)

Country Status (2)

Families Citing this family (9)

Family Cites Families (9)

• 2001
  • 2001-02-09 EP EP01918163A patent/EP1357954A1/de not_active Withdrawn
  • 2001-02-09 WO PCT/US2001/004524 patent/WO2002064183A1/en not_active Application Discontinuation

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events