METHODS FOR COATING SURFACES WITH ANTIMICROBIAL AGENTS

Abstract

Disclosed are methods for coating or impregnating a surface with an antimicrobial agent that involve contacting the surface with a composition that includes an antimicrobial agent and a solvent, and curing the surface by applying heat. Also disclosed are methods for reducing the risk of development or progression of an infection in a subject in need of a medical device, that involve coating or impregnating a surface of the medical device with an antimicrobial agent and then curing the surface by applying heat, wherein the risk of development or progression of an infection is reduced.

Claims

1. A method for coating or impregnating a non-organic surface with minocyline and rifampin, comprising: a) combining the minocycline and rifampin in a solvent to form a mixture and heating the mixture sufficient to dissolve the minocycline and rifampin in the solvent to form an antimicrobial composition; b) coating the non-organic surface with the antimicrobial composition; c) curing the coated non-organic surface at a temperature that is at least about 40 C. to remove at least some of the solvent, and wherein the antimicrobial agent coats or impregnates the surface; d) drying the impregnated surface; and e) heating the dried surface at a temperature of at least 50 C. for at least 12 hours to provide the coated or impregnated non-organic surface.

2. The method of claim 1, further defined as a method for coating or impregnating a medical device with an antimicrobial agent, wherein the non-organic surface is the surface of a medical device.

3. The method of claim 2, wherein the medical device is an endotracheal tube, a tracheotomy tube, chest tube, a vascular catheter, an urinary catheter, a nephrostomy tube, a biliary stent, a peritoneal catheter, an epidural catheter, a central nervous system catheter, an orthopedic device, a prosthetic valve, a gastric tube, an intestinal tube, or drug-delivery implant.

4.-5. (canceled)

6. The method of claim 1, wherein the non-organic surface is a surface of a glove, a condom, a gown, hospital equipment, a table, a container, a countertop, a floor, a bag, a suture, a device used in food-processing, a sponge, or a mop.

7.-32. (canceled)

33. The method of claim 1, wherein the surface comprises a micropore or a nanopore.

34. The method of claim 1, wherein heating the surface is carried out at a temperature that is 50 C. to 100 C.

35. The method of claim 34, wherein heating the surface is carried out at a temperature that is 50 C. to 80 C.

36. The method of claim 35, wherein heating the surface is carried out at a temperature that is 50 C. to 60 C.

37. (canceled)

38. The method of claim 1, wherein the surface is heated for about 12 hours to about 96 hours.

39. The method of claim 38, wherein the surface is heated for about 24 hours to about 72 hours.

40. (canceled)

41. The method of claim 1, further comprises washing the surface after heating the surface.

42. The method of claim 41, wherein washing is further defined as contacting the surface with a composition comprising a detergent and water.

43. The method of claim 41, further comprising heating the surface at a temperature of at least 40 C. after washing the surface.

44. The method of claim 43, wherein the surface is heated for at least 1 hour to at least 96 hours.

45. The method of claim 44, wherein the surface is heated for at least 24 hours to at least 72 hours.

46. The method of claim 1, wherein the surface is composed of a polymer or silicone.

47. The method of claim 46, wherein the polymer is polyvinyl chloride, polyurethane, polyethylene, silastic elastomers, polytetrafluoroethylene, dacron, collodion, carboethane or nylon.

48. The method of claim 2, further comprising packaging the medical device in a container.

49. The method of claim 2, further comprising sterilizing the medical device.

50. (canceled)

51. A method for reducing the risk of development or progression of an infection in a subject in need of a medical device, comprising coating or impregnating a surface of the medical device with an antimicrobial agent in accordance with the method of claim 1 prior to contacting the medical device with the subject, wherein the risk of development or progression of an infection is reduced.

52. The method of claim 51, wherein the subject is a human.

53. The method of claim 51, wherein the antimicrobial agent is further defined as an antibiotic, an antiseptic, an antiviral agent, an antifungal agent, or a disinfectant.

54. The method of claim 53, wherein the antimicrobial agent is an antibiotic.

55.-67. (canceled)

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0053] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present invention. The invention may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

[0054] FIG. 1: Heating reduces Gendine leaching.

[0055] FIG. 2: Heating reduces bacteria (Pseudomonas aerugenosa) adherence.

[0056] FIG. 3: Zones of inhibitions formed by Gendine coated PVC endotracheal tubes in methicillin resistant Staphylococcus aureus.

[0057] FIG. 4: Effects of heating on the leaching of Gendine from coated polyurethane CVC.

[0058] FIG. 5: Adherence of bacteria to polyurethane CVC surfaces.

[0059] FIG. 6: Zones of inhibitions formed by Gendine coated cook polyurethane catheters in methicillin resistant Staphylococcus aureus.

[0060] FIG. 7: Zones of inhibitions formed by Gendine coated cook polyurethane catheters in Pseudamonas aeruginosa.

[0061] FIG. 8: Zones of inhibitions formed by Gendine coated cook polyurethane catheters Candida parapsilosis.

[0062] FIG. 9: Effects of heating on the leaching of Gendine from coated cook silicone urinary catheters.

[0063] FIG. 10: Adherence of bacteria to silicone urinary catheter surfaces.

[0064] FIG. 11: Zones of inhibitions formed by Gendine coated cook silicone urinary catheters in vancomycin resistant Enterococcus (VRE).

[0065] FIG. 12: Zones of inhibitions formed by Gendine coated cook silicone urinary catheters in E. coli.

[0066] FIG. 13: Zones of inhibitions formed by Gendine coated cook silicone urinary catheters in Candida parapsilosis.

[0067] FIG. 14. Durability of Spectrum (unheated Mino-Rifampin catheters) compared to heated mino-rifampin catheters over 12 weekstested against MRSA 4798.

[0068] FIG. 15. Durability of Spectrum compared to heated Mino-Rifampin catheters over 12 weekstested against S. malto 5075.

[0069] FIG. 16. Durability of Spectrum compared to heated Mino-Rifampin catheters over 12 weekstested against S. malto 4807.

[0070] FIG. 17. Durability of Spectrum compared to heated Mino-Rifampin catheters over 12 weekstested against S. malto 4709.

[0071] FIG. 18. Adherent of biofilm to coated polymers.

DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0072] The present invention provides for novel methods of coating or impregnating a surface with an antimicrobial agent. By way of example, the inventors have found that coating a medical device with a composition that includes an antimicrobial agent and a solvent following by curing the medical device at a temperature of greater than 40 C. results in a decrease in staining potential of the medical device, decrease in leaching of the antimicrobial into tissue, and prolonged antimicrobial efficacy. Medical devices, such as indwelling catheters, are used routinely in hospitals on a diverse group of patients. A common cause of failure of these medical devices is infection. Pathogens often attach to and proliferate in such devices and eventually invade the patient leading to nosocomial infections. Microorganisms usually migrate along the surfaces of devices to invade sterile environments, such as the bronchoalveolar space leading to pneumonia, the bloodstream leading to bacteremia, or the urinary bladder leading to urinary tract infections.

[0073] For example, in certain embodiments, the present invention relates to the use of antiseptic compositions with broad-spectrum activity against various nosocomial microorganisms, including resistant bacteria and fungi. For example, the antiseptic compositions are effective against resistant staphylococci, vancomycin-resistant enterococci, resistant Pseudomonas aeruginosa and Candida species. These antiseptics also have unique properties that enable penetration/impregnation of various polymers, such as polyvinyl chloride, polyethylene, silastic elastomers, polytetrafluoroethylene, dacron, collodion, carboethane, nylon, polymers used in the formation of endotracheal tubes, silicone and polyurethane polymers used in the formation of vascular catheters and surgical silk sutures. Thus, they are suitable for coating a wide range of device surfaces.

[0074] The present invention provides safe antimicrobial-treated surfaces wherein the coated or impregnated surface has a durability that may last through the life-span of the device. For example, with regard to medical devices, this significantly decreases patient mortality and morbidity associated with the various nosocomial infections such as nosocomial pneumonias, nosocomial bacteremias, nosocomial urinary tract infections and nosocomial surgical wound infections.

A. ANTIMICROBIAL AGENTS

[0075] For the purposes of this disclosure, the phrase effective amount or therapeutically effective amount is defined as a dosage sufficient to induce a microbicidal or microbistatic effect upon the microbes contacted by the composition on a surface.

[0076] In some embodiments of the invention, the antimicrobial agent is an antibacterial agent. While any antibacterial agent may be used in the preparation of the instant antimicrobial solutions, some non-limiting exemplary antibacterial agent(s) include those classified as aminoglycosides, beta lactams, quinolones or fluoroquinolones, macrolides, sulfonamides, sulfamethaxozoles, tetracyclines, streptogramins, oxazolidinones (such as linezolid), clindamycins, lincomycins, rifamycins, glycopeptides, polymxins, lipo-peptide antibiotics, as well as pharmacologically acceptable sodium salts, pharmacologically acceptable calcium salts, pharmacologically acceptable potassium salts, lipid formulations, derivatives and/or analogs of the above.

[0077] Each of these classes of antibacterial agents have different mechanisms of action and are represented by several antibiotics a discussion of which is presented below. However, the skilled artisan will recognize that the invention is in no way limited to the agents set forth here and that these agents are described merely as examples.

[0078] The aminoglycosides are bactericidal antibiotics that bind to the 30S ribosome and inhibit bacterial protein synthesis. They are typically active against aerobic gram-negative bacilli and staphylococci. Exemplary aminoglycosides that may be used in some specific aspects of the invention include amikacin, kanamycin, gentamicin, tobramycin, or netilmicin.

[0079] Beta lactams are a class of antibacterials that inhibit bacterial cell wall synthesis. A majority of the clinically useful beta-lactams belong to either the penicillin group (penam) or cephalosporin (cephem) groups. The beta-lactams also include the carbapenems (e.g., imipenem), and monobactams (e.g., aztreonam). Inhibitors of beta-lactamase such as clavulanic acid and its derivatives are also included in this category.

[0080] Non-limiting examples of the penicillin group of antibiotics that may be used in the solutions of the present invention include amoxicillin, ampicillin, benzathine penicillin G, carbenicillin, cloxacillin, dicloxacillin, piperacillin, or ticarcillin, etc. Examples of cephalosporins include ceftiofur, ceftiofur sodium, cefazolin, cefaclor, ceftibuten, ceftizoxime, cefoperazone, cefuroxime, cefprozil, ceftazidime, cefotaxime, cefadroxil, cephalexin, cefamandole, cefepime, cefdinir, cefriaxone, cefixime, cefpodoximeproxetil, cephapirin, cefoxitin, cefotetan etc. Other examples of beta lactams include mipenem or meropenem which are extremely active parenteral antibiotics with a spectrum against almost all gram-positive and gram-negative organisms, both aerobic and anaerobic and to which Enterococci, B. fragilis, and P. aeruginosa are particularly susceptible.

[0081] Examples of beta lactamase inhibitors include clavulanate, sulbactam, or tazobactam. In some aspects of the present invention, the antibacterial solutions may comprise a combination of at least one beta lactam and at least one beta lactamase inhibitor.

[0082] Macrolide antibiotics are another class of bacteriostatic agents that bind to the 50S subunit of ribosomes and inhibit bacterial protein synthesis. These drugs are active against aerobic and anaerobic gram-positive cocci, with the exception of enterococci, and against gram-negative anaerobes. Exemplary macrolides include erythromycin, azithromycin, clarithromycin.

[0083] Quinolones and fluoroquinolones typically function by their ability to inhibit the activity of DNA gyrase. Examples include nalidixic acid, cinoxacin, trovafloxacin, ofloxacin, levofloxacin, grepafloxacin, trovafloxacin, sparfloxacin, norfloxacin, ciprofloxacin, moxifloxacin and gatifloxacin.

[0084] Sulphonamides are synthetic bacteriostatic antibiotics with a wide spectrum against most gram-positive and many gram-negative organisms. These drugs inhibit multiplication of bacteria by acting as competitive inhibitors of p-aminobenzoic acid in the folic acid metabolism cycle. Examples include mafenide, sulfisoxazole, sulfamethoxazole, and sulfadiazine.

[0085] The tetracycline group of antibiotics include tetracycline derivatives such as tigecycline which is an investigational new drug (IND), minocycline, doxycycline or demeclocycline and analogs such as anhydrotetracycline, chlorotetracycline, or epioxytetracycline. The present inventors have previously shown that minocycline has a higher penetration of the microbial biofilm layer than vancomycin and that EDTA is unique in effectively preventing and dissolving polysaccharide-rich microbial glycocalyx (U.S. Pat. No. 5,362,754).

[0086] The streptogramin class of antibacterial agents is exemplified by quinupristin, dalfopristin or the combination of two streptogramins.

[0087] Drugs of the rifamycin class typically inhibit DNA-dependent RNA polymerase, leading to suppression of RNA synthesis and have a very broad spectrum of activity against most gram-positive and gram-negative bacteria including Pseudomonas aeruginosa and Mycobacterium species. An exemplary rifamycin is rifampicin.

[0088] Other antibacterial drugs are glycopeptides such as vancomycin, teicoplanin and derivatives thereof. Yet other antibacterial drugs are the polymyxins which are exemplified by colistin.

[0089] In addition to these several other antibacterial agents such as prestinomycin, chloramphenicol, trimethoprim, fusidic acid, metronidazole, bacitracin, spectinomycin, nitrofurantion, daptomycin or other leptopeptides, oritavancin, dalbavancin, ramoplamin, ketolide etc. may be used in preparing the compositions described herein. Of these, metronidazole is active only against protozoa, such as Giardia lamblia, Entamoeba histolytica and Trichomonas vaginalis, and strictly anaerobic bacteria. Spectinomycin, is a bacteriostatic antibiotic that binds to the 30S subunit of the ribosome, thus inhibiting bacterial protein synthesis and nitrofurantoin is used orally for the treatment or prophylaxis of UTI as it is active against Escherichia coli, Klebsiella-Enterobacter species, staphylococci, and enterococci.

[0090] In other embodiments, the antimicrobial agent is an antifungal agent. Some exemplary classes of antifungal agents include imidazoles or triazoles such as clotrimazole, miconazole, ketoconazole, econazole, butoconazole, omoconazole, oxiconazole, terconazole, itraconazole, fluconazole, voriconazole (UK 109,496), posaconazole, ravuconazole or flutrimazole; the polyene antifungals such as amphotericin B, liposomal amphoterecin B, natamycin, nystatin and nystatin lipid formualtions; the cell wall active cyclic lipopeptide antifungals, including the echinocandins such as caspofungin, micafungin, anidulfungin, cilofungin; LY121019; LY303366; the allylamine group of antifungals such as terbinafine. Yet other non-limiting examples of antifungal agents include naftifine, tolnaftate, mediocidin, candicidin, trichomycin, hamycin, aurefungin, ascosin, ayfattin, azacolutin, trichomycin, levorin, heptamycin, candimycin, griseofulvin, BF-796, MTCH 24, BTG-137586, pradimicins (MNS 18184), benanomicin; ambisome; nikkomycin Z; flucytosine, or perimycin.

[0091] In still other embodiments of the invention, the antimicrobial agent is an antiviral agent. Non-limiting examples of antiviral agents include cidofovir, amantadine, rimantadine, acyclovir, gancyclovir, pencyclovir, famciclovir, foscarnet, ribavirin, or valcyclovir. In some embodiments the antimicrobial agent is an innate immune peptide or proteins. Some exemplary classes of innate peptides or proteins are transferrins, lactoferrins, defensins, phospholipases, lysozyme, cathelicidins, serprocidins, bacteriocidal permeability increasing proteins, amphipathic alpha helical peptides, and other synthetic antimicrobial proteins.

[0092] In other embodiments of the invention, the antimicrobial agent is an antiseptic agent. Several antiseptic agents are known in the art and these include a taurinamide derivative, a phenol, a quaternary ammonium surfactant, a chlorine-containing agent, a quinaldinium, a lactone, a dye, a thiosemicarbazone, a quinone, a carbamate, urea, salicylamide, carbanilide, a guanide, an amidine, an imidazoline biocide, acetic acid, benzoic acid, sorbic acid, propionic acid, boric acid, dehydroacetic acid, sulfurous acid, vanillic acid, esters of p-hydroxybenzoic acid, isopropanol, propylene glycol, benzyl alcohol, chlorobutanol, phenylethyl alcohol, 2-bromo-2-nitropropan-1,3-diol, formaldehyde, glutaraldehyde, calcium hypochlorite, potassium hypochlorite, sodium hypochlorite, iodine (in various solvents), povidone-iodine, hexamethylenetetramine, noxythiolin, 1-(3-choroallyl)-3,5,7-triazol-azoniaadamantane chloride, taurolidine, taurultam, N(5-nitro-2-furfurylidene)-1-amino-hydantoin, 5-nitro-2-furaldehyde semicarbazone, 3,4,4-trichlorocarbanilide, 3,4,5-tribromosalicylanilide, 3-trifluoromethyl-4,4-dichlorocarbanilide, 8-hydroxyquinoline, 1-cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-(1-piperazinyl)-3-quinolinecarboxylic acid, 1,4-dihydro-1-ethyl-6-fluoro-4-oxo-7-(1-piperazinyl)-3-quinolinecarboxylic acid, hydrogen peroxide, peracetic acid, phenol, sodium oxychlorosene, parachlorometaxylenol, 2,4,4-trichloro-2-hydroxydiphenol, thymol, chlorhexidine, benzalkonium chloride, cetylpyridinium chloride, silver sulfadiazine, or silver nitrate.

B. SOLVENTS

[0093] Non-limiting examples of solvents are set forth in Table 1.

TABLE-US-00001 TABLE 1 Examples of Solvents Name Structure water HOH methanol CH.sub.3OH ethanol CH.sub.3CH.sub.2OH 1-propanol CH.sub.3CH.sub.2CH.sub.2OH 1-butanol CH.sub.3CH.sub.2CH.sub.2CH.sub.2OH formic acid [00001] acetic acid [00002] formamide [00003] acetone [00004] tetrahydrofuran (THF) [00005] methyl ethyl ketone [00006] ethyl acetate [00007] acetonitrile H.sub.3CCN N,N-dimethylformamide (DMF) [00008] diemthyl sulfoxide (DMSO) [00009] hexane CH.sub.3(CH.sub.2).sub.4CH.sub.3 benzene [00010] diethyl ether CH.sub.3CH.sub.2OCH.sub.2CH.sub.3 methylene chloride CH.sub.2Cl.sub.2 carbon tetrachloride CCl.sub.4

C. DYES

[0094] In some embodiments of the present invention, the surface is contacted with a composition that includes a dye. A dye is defined herein to refer to an agent that is used to impart color. The dye may or may not have antimicrobial activity. In some embodiments, the composition includes a dye and a basic reagent. The dye may or may not be bonded to the basic reagent. The dye may be ionically or covalently bonded to the basic reagent. The dye bonded to the basic reagent may or may not have antimicrobial activity

[0095] The dye may be obtained from any source known to those of ordinary skill in the art. The dye may be obtained, for example, from natural sources, from commercial sources, or may be chemically synthesized.

[0096] The dyes that may be used to synthesize certain antiseptic compounds of the invention include but are not limited to, gentian, or crystal violet, ethyl violet, brilliant green, etc., and the FD&C dyes such as Blue No. 1 and Green No. 3. In addition, other dyes include the following FD&C and D&C colors: (1) monoazo dyes such as, but not limited to, FD&C Yellow No. 5, FD&C Yellow No. 6, (2) diazo dyes such as, but not limited to, D&C Red No. 17, (3) indigoid dyes such as, but not limited to, FD&C Blue No. 2, (4) xanthene (Fluorescein) dyes such as, but not limited to, FD&C Red No. 3, (5) anthraquinone dyes such as, but not limited to, D&C Green No. 6, (6) quinoline dyes such as, but not limited to, D&C Yellow No. 1. An extensive list of dyes and stains that may be employed is also provided in Table 1.

TABLE-US-00002 TABLE 1 The Color Index (C.I.) Number and/or Chemical Abstracts Service Registry CAS) Number for Selected Dyes and Stains: No. C.I. # CAS # 1 15670 2092-55-9 2 26370 3071-73-6 3 20460 5850-35-1 4 62130 2666-17-3 5 61585 4474-24-2 6 26360 3351-05-1 7 62058 6397-02-0 8 42685 3244-88-0 9 61580 6408-57-7 10 15575 5850-86-2 11 22870 15792-50-4 12 18050 3734-67-6 13 14900 4787-93-3 14 18070 12167-45-2 15 22890 10169-02-5 16 23635 6459-94-5 17 18800 6408-31-7 18 18055 4321-69-1 19 18965 6359-98-4 20 18900 6359-91-7 21 25135 13390-47-1 22 22910 6375-5-9 23 18850 6359-88-2 24 46005:1 494-38-2 25 8048-52-0 26 58000 72-48-0 27 3952-78-1 28 61710 6408-63-5 29 42750 30586-13-1 30 569-58-4 31 52417-22-8 32 520-10-5 33 48035 3056-93-7 34 4431-00-9 35 50090 25360-72-9 36 52010 531-55-5 37 61111 12217-43-5 38 42500 569-61-9 39 11460 42373-04-6 40 23500 992-59-6 41 298-95-3 42 21010 5421-66-9 43 1871-22-3 44 28440 2519-30-4 45 42660 6104-59-2 46 27290 5413-75-2 47 24890 3051-11-4 48 76-60-8 49 115-40-2 50 115-39-9 51 65005 1328-24-1 52 62055 6408-78-2 53 62125 6424-85-7 54 63010 2861-02-1 55 13390 3861-73-2 56 26400 3529-01-9 57 15706 12392-64-2 58 61570 4403-90-1 59 62560 4430-16-4 60 26550 8003-88-1 61 18745 10127-27-2 62 14710 5858-39-9 63 17045 6360-07-2 64 15620 1658-56-6 65 18110 6844-74-2 66 26900 6406-56-0 67 18125 10130-48-0 68 42650 4129-84-4 69 18835 6359-85-9 70 18890 6359-90-6 71 18950 6372-96-9 72 14170 6408-90-8 73 13900 10343-58-5 74 46025 135-49-9 75 12840 61968-76-1 76 63615 1324-21-6 77 58005 130-22-3 78 14025 584-42-9 79 42080 3486-30-4 80 16185 915-67-3 81 42780 82 1668-00-4 83 41000 2465-27-2 84 43810 13186-45-3 85 52005 531-53-3 86 51004 33203-82-6 87 11075 94233-04-2 88 42510 632-99-5 89 48055 4208-80-4 90 26905 4196-99-0 91 2315-97-1 92 21000 10114-58-6 93 16180 5858-33-3 94 42655 6104-58-1 95 81029-05-2 96 42040 633-03-4 97 102185-52-4 98 62625-32-5 99 62625-30-3 100 62625-28-9 101 14337-53-2 102 76-59-5 103 40070-59-5 104 3147-14-6 105 24410 2610-05-1 106 43825 1667-99-8 107 16575 548-80-1 108 43820 3564-18-9 109 24895 2870-32-8 110 18972 50662-99-2 111 596-27-0 112 2303-01-7 113 1733-12-6 114 10510-54-0 115 15970 1934-20-9 116 15391-59-0 117 76-54-0 118 4727-50-8 119 54-88-6 120 6473-13-8 121 23655 6420-03-7 122 25380 2829-43-8 123 27905 5489-77-0 124 13950 10190-68-8 125 29025 3214-47-9 126 64500 2475-45-8 127 61500 2475-44-7 128 1005 730-40-5 129 31482-56-1 130 11115 3180-81-2 131 11855 2832-40-8 132 26090 6300-37-4 133 45400 548-24-3 134 45380 548-26-5 135 15086-94-9 136 14640 3564-14-5 137 42090 3844-45-9 138 45430:2 15905-32-5 139 45386 6359-05-3 140 76058-33-8 141 23860 314-13-6 142 11160 97-56-3 143 13015 2706-28-7 144 11285 6416-57-5 145 45350:1 2321-07-5 146 596-09-8 147 3326-34-9 148 51030 1562-85-2 149 1634-82-8 150 3737-95-9 151 165660-27-5 152 16574-43-9 153 34722-90-2 154 617-19-6 155 51050 1562-90-9 156 4430-20-0 157 14720 3567-69-9 158 16570 4197-07-3 159 11270 532-82-1 160 18105 17681-50-4 161 22120 573-58-0 162 2411-89-4 163 62625-31-4 164 62625-29-0 165 41830-80-2 166 42555 548-62-9 167 45370:1 596-03-2 168 620-45-1 169 45425:1 31395-16-1 170 73688-85-4 171 34140 4399-55-7 172 29160 3441-14-3 173 28160 2610-11-9 174 13920 10130-29-7 175 19556 6537-66-2 176 36900 6409-90-1 177 61505 2475-46-9 178 11080 2581-69-3 179 26080 6253-10-7 180 11110 2872-52-8 181 11130 2734-52-3 182 12790 6439-53-8 183 518-82-1 184 56360-46-4 185 45380:2 15086-94-9 186 14645 1787-61-7 187 18760 3618-63-1 188 45430 568-63-8 189 1239-45-8 190 62758-12-7 191 42600 2390-59-2 192 37190 64071-86-9 193 42053 2353-45-9 194 12010 6535-42-8 195 18820 6359-82-6 196 45350 518-47-8 197 3326-32-7 198 51649-83-3 199 42085 4680-78-8 200 75290 517-28-2 201 90-33-5 202 73000 482-89-3 203 73015 860-22-0 204 12210 4569-88-4 205 11050 2869-83-2 206 44090 3087-16-9 207 42000 2437-29-8 208 13065 587-98-4 209 52041 2516-05-4 210 45385 23391-49-3 211 13025 547-58-0 212 32469-43-5 213 14855 3624-68-8 214 11335 6247-27-4 215 11880 6370-46-3 216 11300 6232-53-7 217 26520 3564-27-0 218 18735 1934-24-3 219 14010 6054-99-5 220 44530 5715-76-4 221 11350 131-22-6 222 16255 2611-82-7 223 52030 6586-05-6 224 7385-67-3 225 74-39-5 226 60760 6409-77-4 227 26120 4477-79-6 228 16230 1936-15-8 229 15705 2538-85-4 230 19010 10127-05-6 231 42045 129-17-9 232 34487-61-1 233 101-75-7 234 11800 1689-82-3 235 45410 18472-87-2 236 16680 1058-92-0 237 27190 6226-78-4 238 49000 30113-37-2 239 16593-81-0 240 85531-30-2 241 45005 92-32-0 242 58500 81-61-8 243 47000 8003-22-3 244 20505 17095-24-8 245 61205 13324-20-4 246 17908 25489-36-5 247 635-78-9 248 45170 81-88-9 249 45160 989-38-8 250 45440 632-69-9 251 50240 477-73-6 252 61552 6994-46-3 253 7423-31-6 254 3599-32-4 255 146-68-9 256 42095 5141-20-8 257 42000:1 510-13-4 258 129-16-8 259 52015 61-73-4 260 50206 4569-86-2 261 42590 7114-03-6 262 13020 493-52-7 263 11020 60-11-7 264 20110 3564-15-6 265 11875 6247-28-5 266 13250 3618-62-0 267 14030 2243-76-7 268 26560 6406-37-7 269 6408-91-9 270 14045 6470-98-0 271 20470 1064-48-8 272 50040 553-24-2 273 42520 3248-91-7 274 51180 3625-57-8 275 14890 5423-07-4 276 56431-61-9 277 61555 2646-15-3 278 26125 1320-06-5 279 15510 633-96-5 280 15711 5610-64-0 281 12070 6410-10-2 282 143-74-8 283 11000 60-09-3 284 16201-96-0 285 975-17-7 286 2768-90-3 287 27195 6226-79-5 288 67627-18-3 289 58205 (75410) 81-54-9 290 115-41-3 291 45010 2150-48-3 292 117-92-0 293 58050 81-64-1 294 47005 8004-92-0 295 61211 12236-82-7 296 17757 12225-82-1 297 61200 2580-78-1 298 123333-76-6 299 45170:1 509-34-2 300 13161-28-9 301 43800 603-45-2 302 61554 17354-14-2 303 61565 128-80-3 304 12055 842-07-9 305 12140 3118-97-6 306 26105 85-83-6 307 11920 2051-85-6 308 123359-42-2 309 23647-14-5 310 45100 3520-42-1 311 19140 1934-21-0 312 108321-10-4 313 62637-91-6 314 6262-21-1 315 632-73-5 316 42798-98-1 317 19540 1829-00-1 318 52000 78338-22-4 319 81012-93-3 320 123359-43-3 321 12120 2425-85-6 322 23850 72-57-1 323 44045 2580-56-5 324 42595 2390-60-5 325 125-31-5 326 16150 3761-53-3 327 135-52-4 328 26100 85-86-9 329 26150 4197-25-5 330 26050 6368-72-5 331 68504-35-8 332 123333-78-8 333 45220 5873-16-5 334 4430-25-5 335 1301-20-8 336 123333-63-1 337 386-17-4 338 4430-24-4 339 1719-71-7 340 49005 2390-54-7 341 76-61-9 342 125-20-2 343 52040 92-31-9 344 14270 547-57-9 345 14541-90-3 346 44040 2185-86-6 347 45190 6252-76-2 348 63721-83-5 349 14936-97-1 350

D. BASIC REAGENTS

[0097] Some embodiments of the present invention involve contacting a surface, such as a medical device, with a composition that includes a basic reagent. Any basic reagent known to those of ordinary skill in the art is contemplated. For example, the basic reagents can be alkyl and aryl oxides, thiols, sulfides, phosphorous, aliphatic and aromatic amines, guanidines and halides such as F.sup., Br.sup. and I.sup.. Some examples of the basic reagents that can be used include phenoxide antiseptics (such as clofoctol, chloroxylenol, triclosan) or guanidium compounds (such as chlorhexidine, alexidine, hexamidine) or bipyridines (such as octenidines).

[0098] Other examples include a guanidium compound, a biguanide, a bipyridine, a phenoxide antiseptic, an alkyl oxide, an aryl oxide, a thiol, a halide, an aliphatic amine, or an aromatic amine. Non-limiting examples of guanidium compounds include chlorhexidine, alexidine, hexamidine. In other specific embodiments, the basic reagent is a bipyridine. One example of a bipyridine is octenidine. In yet other aspects, the basic reagent is a phenoxide antiseptic.

E. ANTISEPTICS

[0099] In some embodiments set forth herein, the antimicrobial is an antiseptic. The composition that includes an antiseptic agent may be applied to the surface by any method known to those of ordinary skill in the art. For example, if the surface is a surface of a medical device, the device may be immersed in the composition, or the composition may be painted or sprayed onto the device. The composition may include a dye, as set forth above. The self-impregnating property of the dyes such as for example, the triarylmethane dyes, removes the need for another binding agent. This is another feature of the composition provided by this invention which is a considerable improvement over certain other known compositions. Certain previously known compositions require other impregnating/coating agents and/or must typically be extruded into the device as it is made. Both these methods are time consuming and involve additional steps and techniques.

[0100] For example, one method of coating devices first requires application or absorption of a layer of surfactant, such as tridodecylmethyl ammonium chloride (TDMAC) followed by the antibiotic coating layer, to the surface of the medical device. Another method used to coat surfaces of medical devices with antibiotics involves first coating the selected surfaces with benzalkonium chloride followed by ionic bonding of the antibiotic composition (Solomon and Sherertz, 1987; U.S. Pat. No. 4,442,133). Other methods of coating surfaces of medical devices with antibiotics are taught in U.S. Pat. No. 4,895,566 (a medical device substrate carrying a negatively charged group having a pH of less than 6 and a cationic antibiotic bound to the negatively charged group); U.S. Pat. No. 4,917,686 (antibiotics are dissolved in a swelling agent which is absorbed into the matrix of the surface material of the medical device); U.S. Pat. No. 4,107,121 (constructing the medical device with ionogenic hydrogels, which thereafter absorb or ionically bind antibiotics); U.S. Pat. No. 5,013,306 (laminating an antibiotic to a polymeric surface layer of a medical device); and U.S. Pat. No. 4,952,419 (applying a film of silicone oil to the surface of an implant and then contacting the silicone film bearing surface with antibiotic powders). Furthermore, most of the methods previously employed to coat the surfaces of medical devices use antibiotics such as tetracyclines, penicillins, cephalosporins and the beta-lactam antibiotics. The main drawback with antibiotics is the emergence of resistant strains.

[0101] In certain embodiments, antiseptic derivative compounds with broad-spectrum antiseptic activity against bacteria and fungi including nosocomial and multidrug-resistant varieties may be used to impregnate, bind, coat, adhere and/or attach to various device surfaces without the assistance of impregnating vehicles such as tridodecylmethylammonium chloride (TDMAC). Furthermore, antiseptic compounds of the invention also have an extended antimicrobial efficacy that can cover the life of the device.

[0102] One example of the a broad-spectrum antiseptic is a composition that includes a combination of gentian violet and chlorhexidine (Gendine). Gentian violet, on its own, is a good impregnating triarylmethane dye. Bhatnager et al., 1993 have shown in an in vitro study that gentian violet alone can be used to impregnate the surface of CSF silicone shunts and prevent the colonization of S. epidermis on these surfaces. However, after impregnating the surfaces of various polymers, including polyvinylchloride, gentian violet on its own has no activity against Pseudomonas aeruginosa, which is the second most common cause of nosocomial pneumonia and the third most common cause of nosocomial urinary tract infections. Antiseptics such as chlorhexidine cannot attach on their own onto the surfaces of polyvinylchloride tubes or silicone catheters and silk sutures. They require an impregnating vehicle. Furthermore, on their own they are not highly active against Pseudomonas aeruginosa. On the other hand, upon the binding of gentian violet with chlorhexidine, the new antiseptic agent synthesized, is a potent and effective broad-spectrum antiseptic and has the additional ability to coat/impregnate various device surfaces. Gendine is unique in its ability to impregnate various device polymers, such as polyvinylchloride used in the formation of endotracheal tubes, silicone and polyurethane polymers used in the formation of vascular, as well as peritoneal, epidural, urinary and intraventricular catheters. In addition, gendine is able to impregnate the silk sutures used in surgical wounds.

[0103] Compositions with antiseptic properties that are specifically contemplated for use in the invention include, but are not limited to Gendine, Genlenol and Genfoctol.

F. MICROORGANISMS

[0104] In some embodiments, the methods set forth herein pertain to methods of reducing the risk of development or progression of an infection in a subject. For example, the subject may be a subject in need of a medical device. The infection to be prevented may be, for example bacteremia, pneumonia, meningitis, osteomyelitis, endocarditis, sinusitis, arthritis, urinary tract infections, tetanus, gangrene, colitis, acute gastroenteritis, bronchitis, an abscess, an opportunistic infection, or a nosocomial infection. Examples of bacterial pathogens include Gram-positive cocci such as Staphylococcus aureus, coagulase negative staphylocci such as Staphylococcus epidermis, Streptococcus pyogenes (group A), Streptococcus spp. (viridans group), Streptococcus agalactiae (group B), S. bovis, Streptococcus (anaerobic species), Streptococcus pneumoniae, and Enterococcus spp.; Gram-negative cocci such as Neisseria gonorrhoeae, Neisseria meningitidis, and Branhamella catarrhalis; Gram-positive bacilli such as Bacillus anthracis, Corynebacterium diphtherias and Corynebacterium species which are diptheroids (aerobic and anerobic), Listeria monocytogenes, Clostridium tetani, Clostridium difficile, Escherichia coli, Enterobacter species, Proteus mirablis and other spp., Pseudomonas aeruginosa, Klebsiella pneumoniae, Salmonella, Shigella, Serratia, and Campylobacterjejuni. The antibiotic resistant bacteria that can be killed by the antiseptic coated devices of the present invention include Staphylococci (methicillin-resistant strains), vancomycin-resistant enterococci (Enterococcus faecium), and resistant Pseudomonas aeruginosa.

[0105] Fungal infections may have cutaneous, subcutaneous, or systemic manifestations. Superficial mycoses include tinea capitis, tinea corporis, tinea pedis, onychomycosis, perionychomycosis, pityriasis versicolor, oral thrush, and other candidoses such as vaginal, respiratory tract, biliary, eosophageal, and urinary tract candidoses. Systemic mycoses include systemic and mucocutaneous candidosis, cryptococcosis, aspergillosis, mucormycosis (phycomycosis), paracoccidioidomycosis, North American blastomycosis, histoplasmosis, coccidioidomycosis, and sporotrichosis. Fungal infections include opportunistic fungal infections, particularly in immunocompromised patients such as those with AIDS. Fungal infections contribute to meningitis and pulmonary or respiratory tract diseases.

[0106] Other pathogenic organisms include dermatophytes (Microsporum canis and other M. spp.; and Trichophyton spp. such as T. rubrum, and T. mentagrophytes), yeasts (e.g., Candida albicans, C. Parapsilosis, C. glabrata, C. Tropicalis, or other Candida species including drug resistant Candida species), Torulopsis glabrata, Epidermophytonfloccosum, Malassezia fuurfur (Pityropsporon orbiculare, or P. ovale), Cryptococcus neoformans, Aspergillus fumigatus, and other Aspergillus spp., Zygomycetes (Rhizopus, Mucor), hyalohyphomycosis (Fusarium Spp.), Paracoccidioides brasiliensis, Blastomyces dermatitides, Histoplasma capsulatum, Coccidioides immitis, and Sporothrix schenckii. Other examples include Cladosporium cucumerinum, Epidermophyton floccosum, and Microspermum ypseum.

G. MEDICAL DEVICES

[0107] Non-limiting examples of medical devices are set forth herein. These include vascular devices such as grafts (e.g., abdominal aortic aneurysm grafts, etc.), stents, catheters (including arterial, intravenous, blood pressure, stent graft, etc.), valves (e.g., polymeric or carbon mechanical valves,), embolic protection filters (including distal protection devices), vena cava filters, aneurysm exclusion devices, artificial hearts, cardiac jackets, and heart assist devices (including left ventricle assist devices), implantable defibrillators, electro-stimulation devices and leads (including pacemakers, lead adapters and lead connectors), implanted medical device power supplies, peripheral cardiovascular devices, atrial septal defect closures, left atrial appendage filters, valve annuloplasty devices, mitral valve repair devices, vascular intervention devices, ventricular assist pumps, and vascular access devices (including parenteral feeding catheters, vascular access ports, central venous access catheters); surgical devices such as sutures of all types, anastomosis devices (including anastomotic closures), suture anchors, hemostatic barriers, screws, plates, clips, vascular implants, tissue scaffolds, cerebro-spinal fluid shunts, shunts for hydrocephalus, drainage tubes, catheters including thoracic cavity suction drainage catheters, abscess drainage catheters, biliary drainage products, and implantable pumps; orthopedic devices such as joint implants, acetabular cups, patellar buttons, bone repair/augmentation devices, spinal devices (e.g., vertebral disks and the like), bone pins, cartilage repair devices, and artificial tendons; dental devices such as dental implants and dental fracture repair devices; drug delivery devices such as drug delivery pumps, implanted drug infusion tubes, drug infusion catheters, and intravitreal drug delivery devices; ophthalmic devices such as scleral buckles and sponges, glaucoma drain shunts and intraocular lenses; urological devices such as penile devices (e.g., impotence implants), sphincter, urethral, prostate, and bladder devices (e.g., incontinence devices, benign prostate hyperplasia management devices, prostate cancer implants, etc.), urinary catheters including indwelling (Foley) and non-indwelling urinary catheters, and renal devices; synthetic prostheses such as breast prostheses and artificial organs (e.g., pancreas, liver, lungs, heart, etc.); respiratory devices including lung catheters; neurological devices such as neurostimulators, neurological catheters, neurovascular balloon catheters, neuro-aneurysm treatment coils, and neuropatches, splints, nasal tampons, ear wicks, ear drainage tubes, tympanostomy vent tubes, otological strips, laryngectomy tubes, esophageal tubes, esophageal stents, laryngeal stents, salivary bypass tubes, and tracheostomy tubes; oncological implants; and pain management implants.

[0108] Classes of suitable non-implantable devices can include dialysis devices and associated tubing, catheters, membranes, and grafts; autotransfusion devices; vascular and surgical devices including atherectomy catheters, angiographic catheters, intraaortic balloon pumps, intracardiac suction devices, blood pumps, blood oxygenator devices (including tubing and membranes), blood filters, blood temperature monitors, hemoperfusion units, plasmapheresis units, transition sheaths, dialators, intrauterine pressure devices, clot extraction catheters, percutaneous transluminal angioplasty catheters, electrophysiology catheters, breathing circuit connectors, stylets (vascular and non-vascular), coronary guide wires, peripheral guide wires; dialators (e.g., urinary, etc.); surgical instruments (e.g. scalpels and the like); endoscopic devices (such as endoscopic surgical tissue extractors, esophageal stethoscopes); and general medical and medically related devices including blood storage bags, umbilical tape, membranes, gloves, surgical drapes, wound dressings, wound management devices, needles, percutaneous closure devices, transducer protectors, pessary, uterine bleeding patches, PAP brushes, clamps (including bulldog clamps), cannulae, cell culture devices, materials for in vitro diagnostics, chromatographic support materials, infection control devices, colostomy bag attachment devices, birth control devices; disposable temperature probes; and pledgets.

H. EXAMPLES

[0109] The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.

Example 1

Method of Coating Medical Device Results in Superior Properties

Materials and Methods

[0110] Preparation of Pieces.

[0111] Gendine was prepared as described in U.S. Patent App. Pub. No. 20030078242. One-centimeter segments of endotracheal tube (ETT) made of polyvinylchloride (PVC-ETT) and urinary catheter (UC) made of silicone (SilUC) and central venous catheter (CVC) made of polyurethane (CVC-PU) were dipped into the Gendine solution, so that both internal and external surfaces were coated. The pieces were left to dry overnight, at 60 C. The pieces were then washed using mild detergent and de-ionized water to remove any possible loosely attached antiseptic off the surface of the coated segments. Some of the pieces were then left to dry for and additional 48 hours at a temperature of 60 C.

[0112] Absorbance and Quantitation of Leaching.

[0113] To determine leaching of the dye, sets of standards with known concentrations of Gentian violet were made in each biological fluid (bronchoalveolar lavage, plasma, and artificial urine). Absorbances were read for each of the standards and a standard curve was plotted. Absorbances were also read from 200 L biological fluids of each coated segments. Quantitation of leaching (in g/L) was derived from the regression line in the standard curve. Unknown absorbances were read at 550 nm and then figured into the standard curve equation to determine amount of leaching in g/L.

[0114] Adherence Testing.

[0115] The inventors evaluated bacterial adherence to the surface of Gendine-coated and control uncoated segments, as well as that of silver hydrogel urinary catheter and other antiinfective approved CVC, such as the antibiotic-coated CVC (Spectrum, Cook Critical Care, Bloomington, Ind.coated with minocycline and rifampin), CVC coated with chlorhexidine/silver sulfadiazine (Arrow Guard Plus, Arrow, Reading, Pa.) and silver/platinum CVC (Edwards, Edwards Life Sciences, Irvine, Calif.). Six segments of each device were tested per organism. A modification of a previously published method for testing adherence and biofilm formation on silicone disks was used (Chaiban et al., 2005).

[0116] The sterile device segments were placed into sterile 5 mL snap top tubes containing 1 mL of plasma. The tubes were then placed into the incubator for 24 hours at 37 C. The plasma was then removed from the tubes, leaving the pieces inside the tubes, and was replaced with 1 mL of Mueller Hinton Broth (MHB), that was inoculated with bacteria. The inoculum was prepared as follows: Five colonies of freshly subbed bacteria were placed in 50 mLs of MHB (0.5 McFarland, approximately 1.3108 CFU/mL), and used immediately. The tubes were then placed in the incubator for 24 hours at 37 C. The MHB was then removed and replaced with 1 mL of 0.9% saline solution and the tube was placed in the 37 C. incubator for 30 minutes as a washing step. The catheter segments were then removed from the washing saline, and placed into sterile 15 mL tubes containing 5 mLs of 0.9% sterile saline solution and sonicated for 15 minutes. After sonication, the tubes containing the catheters were vortexed for 60 seconds. A 100 L volume of the sample was pipetted and spread onto a trypticase soy agar plate with 5% sheep blood (this was the 1:50 dilution). The plates were placed in the incubator for 24 hours, and then the colonies were counted. A value of 100 CFU was used for any plate that had at least 100 counted colonies.

[0117] Zones of Inhibition and Antimicrobial Durability.

[0118] Using a modified Kirby-Bauer method, baseline antimicrobial activity was assessed by measuring the zones of inhibition created by duplicates of impregnated segments that were vertically embedded in Mueller Hinton agar plates coated with one of the following organisms: Methicillin-resistant staphylococcus aureus (MRSA), C. albicans, C. parapsilosis, Vancomycin-resistant Enterococci (VRE), P. aeruginosa, and E. coli. The zones of inhibition were measured and recorded as the diameter (mm) across the center of the embedded segments.

[0119] The antimicrobial durability of Gendine-coated ETT (GND-ETT), Gendine-coated UC (GND-UC) and Gendine-coated CVC (GND-CVC) segments was assessed over time by testing zones of inhibitions of segments soaked in body fluids. The antimicrobial durability of that GND-CVC was compared with other anti-infective CVC described above (Arrow Guard Plus, Edwards and Spectrum). The segments were placed in sterile 50 mL polystyrene tubes (Falcon), containing 10 mL sterile bronchoalveolar lavage (ETT) or urine (UC), or serum (CVC), respectively, and were incubated at 37 C. The 10 mL volume was used to ensure the complete immersion of all of the pieces contained in the tube. Segments were tested in duplicates at weekly intervals, as the soaking fluids were replaced with fresh fluids. Zones of inhibition were determined using the modified Kirby-Bauer method against the same organisms mentioned above.

[0120] Abbreviations.

[0121] MRSA (bacteria)=multidrug-resistant Staphylococcus aureus. PS (bacteria)=Pseudomonas aeruginosa. EC (bacteria)=Escherichia coli. VRE (bacteria)=vancomycin-resistant Enterococcus. CA (fungi)=Candida albicans.

Results

[0122] Endotracheal Tubes (ETT).

[0123] As shown in FIG. 1, heating of the Gendine-coated ETT decreased leaching of the antiseptic dye (Gendine) progressively and substantially (P<0.01). More than three-fold decrease in leaching of Gendine was quantitatively noted after three days of heating (Gendine H3) and by more than two-fold after one day of heating (Gendine H1). Furthermore, heating of the Gendine-coated ETT improved anti-infective anti-adherence effect of the Gendine-coated ETT against multidrug-resistant Pseudomonas aeruginosa as shown in FIG. 2. This improvement was significant (P<0.001) after one day of heating (Gendine H1) and three days of heating (Gendine H3). Heating did not compromise the antimicrobial durability of the Gendine-coated ETT against MRSA through the series of zones of inhibitions performed. This high level of antimicrobial durability was hence maintained as shown in FIG. 3.

[0124] Central Venous Catheters (CVC).

[0125] As shown in FIG. 4 below, heating progressively decreased the leaching of Gendine-coated CVC into the serum (P<0.05). Heating of Gendine-coated CVC for one day (Gendine H1) resulted in almost 1.6-fold decrease in leaching whereas three days of heating (Gendine H3) was associated with 3.5-fold decrease leaching into the serum when compared to unheated Gendine-coated CVC. Furthermore, as shown in FIG. 5, three-day heating of Gendine-coated CVC (Gendine H3) maintained a significantly improved anti-adherence effect of multidrug-resistant bacteria, such as Pseudomonas aeruginosa and MRSA when compared with uncoated CVC or other antimicrobial CVC, such as the spectrum coated with antibiotics (minocycline and rifampin) or Arrow Guard (coated with chlorhexidine and silver sulfadiazine) or the Edwards catheter (with electric silver and platinum ions) (p<0.04). Three day heating (Gendine H3) also maintained a significantly superior anti-adherence effect compared to uncoated CVC with the Spectrum and the Edwards catheter against fungi, (p<0.02) such as Candida albicans and Candida parapsilosis, which are known to be associated with catheter-related candidemia (FIG. 5). In addition, the heating of Gendine-coated CVC (Gendine H1 and Gendine H3) continued to maintain a superior antimicrobial durability against MRSA, multidrug-resistant Pseudomonas aeruginosa and Candida parapsilosis when compared to other antimicrobial catheters, such as the Spectrum, Arrow Guard and Edwards, as shown in FIG. 6, FIG. 7 and FIG. 8.

[0126] Urinary Catheters (UC).

[0127] FIG. 9 shows that heating of Gendine-coated UC (Gendine H1 and Gendine H3) progressively and substantially decreased leaching (P<0.001). The one-day heating (Gendine H1) decreased the leaching of silicone UC by 1.6-fold, whereas the three-day heating (Gendine H3) decreased the leaching into urine of the Gendine by 2.5-fold. Furthermore, as shown below, heating of Gendine-coated UC (Gendine H3-UC) maintained a significantly (P0.025) improved anti-adherence effect, compared to unocated to uncoated urinary catheters or silver-coated UC against two isolates of multidrug-resistant Pseudomonas and two isolates of E. coli (FIG. 10). It should be noted that the silver-coated UC has been shown effective in decreasing bacteruria clinically. In addition, as shown in FIG. 11, FIG. 12 and FIG. 13, heating of Gendine-coated silicone UC maintained a high level of antimicrobial durability over a 56-day period against vancomycin-resistant Enterococci (VRE), resistant E. coli and Candida parapsilosis tested through zones of inhibition and urine.

[0128] Heating of Gendine-coated ETT was associated with a significant decrease in leaching into bronchoalveolar lavage (p<0.05) and a significant improvement in the anti-adherence/antimicrobial activity of Gendine-coated ETT against multidrug-resistant Pseudomonas aeruginosa (p<0.001). The high level of antimicrobial durability of Gendine-coated ETT against methicillin resistant staphylococcus aureus (MRSA) was maintained through heating.

[0129] Heating of Gendine-coated CVC was progressively and substantially associated with decreased leaching of this antiseptic dye into serum while maintaining a significant anti-adherence activity against resistant bacteria and fungi compared to other antimicrobial catheters on the market. Furthermore, the antimicrobial durability of the heated Gendine-coated CVC was maintained in a superior fashion to other antimicrobial catheters placed in serum.

[0130] The heating of silicone UC coated with Gendine was associated with a progressive and substantial decrease of this antiseptic dye into urine while maintaining a significantly superior anti-adherence/antimicrobial activity against Multidrug-resistant gram-negative bacteria, such as Pseudomonas aeruginosa and E. coli compared to the only available anti-infective urinary catheter on the market, which is the silver hydrogel-coated Bard UC. Furthermore the high level of antimicrobial durability of Gendine-coated UC was maintained through heating for up to 56 days against fungi, such AS Candida parapsilosis, multidrug-resistant organisms, such as VRE and E. coli.

[0131] All medical devices coated with antiseptic dyes or antiseptic/antimicrobial agents including.

Example 2

Studies Demonstrating that Heating Enhances the Durability of Antimicrobial Efficacy and Adherence of Biofilm to Minocycline/Rifampin-Coated and Heated Silicone and CVCs

Materials and Method

[0132] Antimicrobial Coating Procedure.

[0133] Constantly stirring, 450 mg of NaOH was dissolved in a beaker containing 45 mL of methanol heated to 45 C. Once completely dissolved, 4.5 g of minocycline was added to the solution in small aliquots over 15 minutes and stirred until completely dissolved. Then, 9 g of rifampin were added to the solution in small aliquots over 15 minutes. Solution was stirred at 45 C. until completely clear. Finally 255 mL of prewarmed (45 C.) butyl acetate was added to the solution and stirred until thoroughly mixed.

[0134] Whole catheters (Cook silicone CVC) were coated for 1 hour at 45 C. Catheters were removed and allowed to dry for 1 day as described below.

[0135] 1 day heatingDried at 60 C. overnight (14-24 hours). Washed twice (ddH.sub.2O, ddH.sub.2O+detergent for 1 minute then dried for an additional 4 hours at 60 C. After drying, the catheters were cut into 0.5 cm segments for in vitro durability and adherence testing. Uncoated catheters and commercially available Spectrum catheters (coated catheters which were not heated) were also tested for comparison.

[0136] Efficacy and Durability of Antiseptic Activity in Coated CVCs.

[0137] Using the modified Kirby-Bauer method, coated catheter segments were vertically inserted in agar plates inoculated with a 0.5 McFarland dilution of microorganisms. Organisms to be tested against included:

[0138] 1. methicillin resistant Staphylococcus aureus (MRSA 4978)clinical isolate

[0139] 2. Stenotrophomonas maltophilia (5075, 4709, 4807)catheter site isolate

[0140] Plates were incubated overnight then zones of inhibition (ZOI) were measured in millimeters (mm). Remaining coated segments were incubated in donor calf serum. Weekly segments were removed for durability testing and the donor calf serum was replaced with fresh.

[0141] Adherence of Biofilm to Coated CVCs.

[0142] Following a modified Kuhn's method, coated catheter segments were incubated for 24 hours in plasma. The plasma was then replaced with 5.010.sup.5 cells in Muller Hinton Broth of either methicillin resistant Staphylococcus aureus (MRSA 4798) or Stenotrophomonas maltophilia (5075, 4709, 4807) and incubated for an additional 24 hours. After incubation, the bacterial innoculum was discarded and segments were washed shaking for 30 minutes in 1 mL of 0.9% sterile saline. The segments were then removed with sterile sticks placed in 5 mL of 0.9% sterile saline and sonicated for 15 minutes. After sonication, 100 L of liquid from each segment was spread onto trypticase soy agar with 5% sheep blood and incubated at 37 C. inverted for 24 hours. Plates were then counted for colony growth.

[0143] Statistical Methods.

[0144] For each bacterium strain, the numbers of viable organisms adhering to the catheter segments indicated by CFU were compared by Kruskal-Wallis test (P<0.05 was regarded statistically significant). If a significant result was detected for the test, a Wilcoxon rank sum tests for the following pairwise comparisons was made: comparing PU-Mino/Rifam (H1=heated) with PU-Spectrum (not heated) and comparing each of them with control, respectively; comparing Silicone-Mino/Rifam (H1=heated) with Silicone-Spectrum (not heated) and comparing each of them with control, respectively. The levels of the post-hoc pairwise comparisons were adjusted using a sequential Bonferroni adjustment to control type I error.

Results

[0145] Table 2 demonstrates results of studies evaluating the efficacy and durability of antiseptic activity in coated CVCs. Spectrum catheters are those coated with minocycline and rifampin without any heating to determine the antimicrobial durability of catheters coated with minocycline and rifampin. Segments of spectrum silicone catheters coated with minocycline and rifampin (without heating) and other silicone catheters coated with minocycline and rifampin that were heated were immersed in serum. Catheter segments were removed at weekly intervals and we determined zones of inhibition (ZOI) against resistant bacteria such as methicillin resistant staphylococci (MRSA) and strenotrophomas maltophilia (S. malta) strains. A ZOI of 10 mm is predictive of in vivo and clinical efficacy. It was found that heating of the catheters treated with Mino/Rifampin increased the efficacy (ZOI10) of coating against MRSA from 28 days to 70 days (Table 2). It was also found that heating of the catheters treated with Mino/Rifampin increased the efficacy (ZOI10) of coating against Stenotrophomonas maltophilia by 1 week.

TABLE-US-00003 TABLE 2 Efficacy and Durability of antiseptic activity in Coated CVCs as determined by zones of inhibition: S. malto S. malto S. malto MRSA 4798 5075 4807 4709 Silicone- Specturm 21-Feb-2006 Baseline 27 27 18 18 20 19 18 18 (Day 0) 28-Feb-2006 Day 7 21 21 0 0 12 12 12 12 7-Mar-2006 Day 14 18 16 0 0 0 0 0 0 14-Mar-2006 Day 21 15 14 0 0 0 0 0 0 21-Mar-2006 Day 28 13 13 0 0 0 0 0 0 28-Mar-2006 Day 35 8 8 0 0 0 0 0 0 4-Apr-2006 Day 42 8 7 0 0 0 0 0 0 11-Apr-2006 Day 49 7 7 0 0 0 0 0 0 18-Apr-2006 Day 56 5 5 0 0 25-Apr-2006 Day 63 0 0 0 0 2-May-2006 Day 70 0 0 0 0 9-May-2006 Day 77 0 0 0 0 Day 84 Silicone-Mino/ Rifampin 1 day heat 21-Feb-2006 Baseline 32 30 20 20 22 21 21 20 (Day 0) 28-Feb-2006 Day 7 25 24 8 7 16 15 15 12 7-Mar-2006 Day 14 22 21 0 0 12 12 8 7 14-Mar-2006 Day 21 21 21 0 0 0 0 0 0 21-Mar-2006 Day 28 20 19 0 0 0 0 0 0 28-Mar-2006 Day 35 14 14 0 0 0 0 0 0 4-Apr-2006 Day 42 10 10 0 0 0 0 0 0 11-Apr-2006 Day 49 10 10 0 0 0 0 0 0 18-Apr-2006 Day 56 10 10 0 0 25-Apr-2006 Day 63 10 10 0 0 2-May-2006 Day 70 10 10 0 0 9-May-2006 Day 77 7 7 0 0 Day 84

[0146] Table 3 demonstrates results of studies pertaining to an evaluation of the adherence of biofilm forming bacteria biofilm to coated CVCs.

[0147] Adherence of biofilm to coated CVC's:

[0148] Following a modified Kuhn's method coated silicone catheter segments were incubated for 24 hours in plasma. The plasma was then replaced with 5.010.sup.5 cells in Muller Hinton Broth of either methicillin resistant Staphylococcus aureus (MRSA 4798) or Stenotrophomonas maltophilia (5075, 4709, 4807) and incubated for an additional 24 hours. After incubation, the bacterial innoculum was discarded and segments were washed by shaking for 30 minutes in 1 mL of 0.9% sterile saline. The segments were then removed with sterile sticks placed in 5 mL of 0.9% sterile saline and sonicated for 15 minutes. After sonication, 100 L of liquid from each segment was spread onto trypticase soy agar with 5% sheep blood and incubated at 37 C inverted for 24 hours. Plates were them counted for colony growth.

[0149] It was found that heating of polyurethane and silicone CVCs increased the inhibition of biofilm forming bacteria when tested against MRSA and S. malto. When tested against MRSA Sil Mino/Rifampin-1 (heated at 60 C. for one day) significantly decreased adherence (p=0.0047) when compared to CVCs coated with minocycline and rifampin without heating. All coated catheters showed significant decrease of adherence of biofilm forming bacteria (p<0.0001) when compared to uncoated control catheters.

TABLE-US-00004 TABLE 3 Adherence of biofilm forming bacteria to coated CVCs: PU Mino/ Control Sil- Control Control Rifampin Silicone- Control Mino/Rifampin PU-Uncoated PU Spectrum 1 day heat Uncoated Sil-Spectrum 1 day heat Plate Plate Plate Plate Plate Plate count Dilution count Dilution count Dilution count Dilution count Dilution count Dilution Tested against adherence of methicillin resistant Staph aureus (MRSA 4798) 1 100 5000 0 0 0 0 100 5000 1 50 0 0 2 100 5000 4 200 1 50 100 5000 1 50 0 0 3 100 5000 5 250 4 200 100 5000 4 200 1 50 4 100 5000 13 650 10 500 100 5000 4 200 4 200 5 100 5000 45 2250 27 1350 100 5000 5 250 0 0 6 100 5000 6 300 46 2300 100 5000 5 250 0 0 7 100 5000 73 3650 14 700 100 5000 6 300 4 200 8 100 5000 49 2450 0 0 100 5000 11 550 4 200 9 100 5000 92 4600 2 100 100 5000 3 150 0 0 10 100 5000 47 2350 5 250 100 5000 31 1550 0 0 Average 5000 1670 545 5000 355 65 St Dev 0.00 1626.04 745.15 0.00 443.13 94.43 Tested against adherence of Stenotrophomas Maltophilia (PS 4807 & 4709) 1 100 5000 0 0 3 150 100 5000 0 0 0 0 2 100 5000 0 0 4 200 100 5000 0 0 0 0 3 100 5000 2 100 22 1100 100 5000 0 0 0 0 4 100 5000 15 750 42 2100 100 5000 1 50 2 100 5 100 5000 100 5000 14 700 100 5000 100 5000 0 0 6 100 5000 67 3350 3 150 100 5000 100 5000 0 0 7 100 5000 100 5000 6 300 100 5000 100 5000 6 300 8 100 5000 100 5000 2 100 100 5000 100 5000 11 550 9 100 5000 100 5000 6 300 100 5000 100 5000 4 200 10 100 5000 3 150 9 450 100 5000 100 5000 7 350 Average 5000 2435 555 5000 3005 150 St Dev 0.00 2415.81 624.26 0.00 2575.57 194.37
1) Results pertaining to adherence of S. maltophilia (PS 4807):

[0150] The Kruskal-Wallis test detected a significant difference for PS4807 CFU among the 3 types of CVCs (.sup.2.sub.(5)=43.97, p<0.0001).

[0151] According to the Wilcoxon rank sum tests, there was no significant difference in CFU between PU Mino/Rifam (H1=heated) and PU-Spectrum (not heated) (p=0.45), but both catheters significantly reduced CFU compared to the control (p<0.0001 for PU Mino/Rifam (H1=heated) vs control; p=0.0059 for PU-Spectrum (not heated) vs control). The CFU of Si-Mino/Rifam (H1=heated) was significantly less than that of Si-Spectrum (not heated) (p=0.022), which in turn was less than that of the control (p<0.0001).

TABLE-US-00005 TABLE 4 S. maltophilia 4807 biofilm CFU among different catheters Catheter Mean Median (Min-Max) N PU-Uncoated 5000 5000 (5000-5000) 10 PU-Spectrum 2435 2050 (0-5000) 10 PU-Mino/Rifam (H1) 555 300 (100-2100) 10 Silicone-Uncoated 5000 5000 (5000-5000) 10 Silicone-Spectrum 240 125 (0-1250) 10 Silicone-Mino/Rifam (H1) 30 0 (0-150) 10 Kruskal-Wallis test: Chi-square statistic with 5 d.f. = 43.97, p < .0001 PU = Cool Polyurethane Spectrum = CVC coated with minocycline & rifampin without heating. Mino/Rifam (H1) = CVC coated with minocycline & rifampin with 1 day heating at 60 C. Comparisons: (PS4807) Catheters: p-value 1. PU-Mino/Rifam (H1) vs Pu-Spectrum 0.45 2. PU-Mino/Rifam (H1) vs PU-Uncoated <.0001 3. PU-Spectrum vs PU-Uncoated 0.0059 1. Si-Mino/Rifam (H1) vs Si-Spectrum 0.022 2. Si-Mino/Rifam (H1) vs Si-Uncoated <.0001 3. Si-Spectrum vs Si-Uncoated <.0001
2) Results Pertaining to Adherence of S. maltophilia (PS 4709):

[0152] The Kruskal-Wallis test detected a significant difference for PS4709 CFU among the 3 types of CVCs (.sup.2.sub.(5)=34.36, p<0.0001).

[0153] According to the Wilcoxon rank sum tests, there was no significant difference in CFU between PU Mino/Rifam (H1=heated) and PU-Spectrum (not heated) (p=0.85), and between a trend towards reducing adherence when we compared Silicone-Mino/Rifam (H1=heated) and to Silicone-Spectrum (coated with minocycline and rifampin but not heated)(p=0.081), respectively. However all the Mino/Rifam coated catheters significantly reduced CFU than their corresponding controls, respectively (Table 4).

TABLE-US-00006 TABLE 5 S. maltophilia 4709, Biofilm CFU among different catheters Catheter Mean Median (Min-Max) N PU-Uncoated 5000 5000 (5000-5000) 10 PU-Spectrum 650 525 (0-2900) 10 PU-Mino/Rifam (H1) 1600 600 (0-5000) 10 1. Silicone-Uncoated 5000 5000 (5000-5000) 10 2. Silicone-Spectrum 3005 5000 (0-5000) 10 3. Silicone-Mino/Rifam (H1) 150 50 (0-550) 10 Kruskal-Wallis test: Chi-square statistic with 5 d.f. = 34.36, p < .0001 Comparisons: (PS4709) Catheters: p-value 1. PU-Mino/Rifam (H1) vs Pu-Spectrum 0.85 2. PU-Mino/Rifam (H1) vs PU-Uncoated 0.0007 3. PU-Spectrum vs PU-Uncoated <.0001 1. Si-Mino/Rifam (H1) vs Si-Spectrum 0.081 2. Si-Mino/Rifam (H1) vs Si-Uncoated <.0001 3. Si-Spectrum vs Si-Uncoated 0.034

3) Results Pertaining to MRSA 4798

[0154] The Kruskal-Wallis test detected a significant difference for MRSA 4798 CFU among the 6 CVCs (.sup.2.sub.(5)=47.86, p<0.0001). According to the Wilcoxon rank sum tests (see Table 6), there was no significant difference in CFU between PU Mino/Rifam (H1=heated) and PU-Spectrum (not heated) ((p=0.088), but both catheters significantly reduced CFU compared to the control (p<0.0001, respectively). The CFU of Si-Mino/Rifam (H1=heated) was significantly less than that of Si-Spectrum (not heated) (p=0.0047), which in turn was less than that of the control (p<0.0001).

TABLE-US-00007 TABLE 6 MRSA 4798 Biofilm CFU among different catheters Catheter Mean Median (Min-Max) N PU-Uncoated 5000 5000 (5000-5000) 10 PU-Spectrum 1670 1450 (0-4600) 10 PU-Mino/Rifam (H1) 545 225 (0-2300) 10 Silicone-Uncoated 5000 5000 (5000-5000) 10 Silicone-Spectrum 355 225 (50-1550) 10 Silicone-Mino/Rifam (H1) 65 0 (0-200) 10 Kruskal-Wallis test: Chi-square statistic with 5 d.f. = 47.86, p < .0001 Comparisons: (MRSA 4798)) Catheters: p-value 1. PU-Mino/Rifam (H1) vs Pu-Spectrum 0.088 2. PU-Mino/Rifam (H1) vs PU-Uncoated <.0001 3. PU-Spectrum vs PU-Uncoated <.0001 4. Si-Mino/Rifam (H1) vs Si-Spectrum 0.0047 5. Si-Mino/Rifam (H1) vs Si-Uncoated <.0001 6. Si-Spectrum vs Si-Uncoated <.0001

[0155] All of the methods disclosed and claimed herein can be executed without undue experimentation in light of the present disclosure. While the of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the methods or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

REFERENCES

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