Medical devices with coatings to reduce biofilm formation and / or growth

Abstract

The present invention relates to a medical device comprising a coating, the coating comprising a polymer Z and a moiety of formula A attached to the medical device, wherein X is an amino group covalently attached to Z or an amino group capable of ionically bonding to free carboxyl groups, wherein the moiety according to formula A is present in the coating in an amount of 0.5 to 30 nmol / cm2, and the polymer Z comprises free carboxyl groups in an amount of 1 to 30 μmol / cm2. [Formula 1] JPEG2026502041000032.jpg19159

Description

[Technical Field]

[0001] The present invention relates to medical devices for reducing the formation and / or growth of biofilms on their surfaces. More specifically, the present invention relates to medical devices having a coating comprising 2-(pyridyldithio)ethylamine (PDEA) or a related compound covalently bonded to a polymer or non-covalently bonded to the coating. The amount of covalently or non-covalently bonded PDEA in the coating may be low and still enable the medical device to reduce the formation and / or growth of biofilms when exposed to microorganisms, such as those in the human body. [Background technology]

[0002] Biofilms are aggregates of microorganisms embedded in a matrix of extracellular polymeric substances that readily attach and grow on surfaces, such as solid surfaces. The microorganisms can be of various types, including bacteria, archaea, protozoa, fungi, and algae. The biofilm structure has been found to provide a natural barrier and protective layer for microorganisms, allowing them to thrive and enhancing their resistance to environmental stresses compared to non-interconnected planktonic organisms. Therefore, biofilm formation appears to be a way for microorganisms to survive exposure to external stressors, such as UV radiation, extreme temperatures and pH, chemicals, disinfectants, and antimicrobial agents. However, the protection conferred to microorganisms and the widespread occurrence of biofilms pose problems in many sectors, including the healthcare sector, where biofilms have been reported to be involved in 80% of chronic infections and 65% of other infectious diseases.

[0003] Biofilms are of particular concern because they can withstand or resist treatment with antimicrobial agents, such as antibiotics, making treatment of infections associated with the presence of biofilms difficult or even impossible. The difficulty in treating biofilm-associated infections with antibiotics has been reported, for example, due to poor antibiotic penetration into biofilms, which can lead to poor diffusion within the biofilm, resulting in low antibiotic concentrations and, as a result, bacterial survival and resistance. Furthermore, when incorporated into biofilms, bacteria undergo a transition from planktonic to sessile growth. This transition results in a decrease in metabolic activity and a decrease in susceptibility to antibiotics. As a result, the concentration of some antibiotics required to kill bacteria embedded in biofilms can be up to 1,000 times higher than the concentration required to kill planktonic bacteria.

[0004] Therefore, biofilm formation is an important cause of antibiotic resistance. Because antibiotics are one of the most important tools for treating bacterial infections and diseases, this means a reduced ability to properly treat patients, which leads to increased morbidity and mortality. Unfortunately, antimicrobial resistance has been increasing for many years, and the World Health Organization (WHO) has recognized it as one of the top 10 global health challenges.

[0005] Biofilm formation and growth frequently occur on the surfaces of medical devices used in modern medicine. Biofilms can form on both intermittently used and indwelling medical devices, such as implants and catheters. For example, catheter-associated urinary tract infections (CAUTIs) are one of the most common healthcare-associated infections affecting many hospitalized patients with urinary catheters. Given the widespread use of medical devices, this represents a major challenge for the healthcare sector, both in terms of human suffering and economic pressure. Therefore, measures to prevent biofilm formation and growth have attracted considerable attention and effort.

[0006] Over the past few years, a number of coating materials have been developed to modify surfaces with the aim of minimizing bacterial and biofilm adhesion, including antimicrobial agents such as small molecules, silver ions, and nanoparticles.

[0007] WO 2006 / 101438 discloses an antimicrobial agent comprising a cysteine ​​compound covalently bound to a substrate, particularly via an S-S spacer molecule. It is stated that the agent has excellent antimicrobial properties and can be used to coat the surface or substrate of various devices, such as medical devices, for the purpose of reducing microbial accumulation and / or growth and / or proliferation and / or viability and / or biofilm formation. Example 16 states that the presence of a cys moiety is essential for the antimicrobial effect. Odeberg et al. (A novel cysteine-linked antibacterial surface coating significantly inhibits bacterial colonization of nasal silicone prongs in a phase one pre-clinical trial, Mater Sci Eng C Mater Biol Appl 2018 Dec 1;93:782-789) disclose a phase 1 first-in-human trial of cysteine ​​compound-coated nasal prongs made by the method disclosed in WO 2006 / 101438. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] International Publication No. 2006 / 101438 [Non-patent literature]

[0009] [Non-Patent Document 1] Odeberg et. al., A novel cysteine-linked antibacterial surface coating significantly inhibits bacterial colonization of nasal silicone prongs in a phase one pre-clinical trial, Mater Sci Eng C Mater Biol Appl 2018 Dec 1;93:782-789 Summary of the Invention [Problem to be solved by the invention]

[0010] While many coatings have proven effective, there remains a need for improved coatings, such as coatings that prevent biofilm formation and / or growth when exposed to microorganisms for several days or longer. It is an object of the present invention to overcome or at least alleviate some of the problems associated with known coatings. It is a further object of the present invention to provide aspects and / or advantages not available with previously known techniques. [Means for solving the problem]

[0011] The present invention relates to a polymer containing free carboxyl groups, the moiety of formula A [ka] This invention is based on the unexpected discovery that a coating containing a compound of Formula A in a quantity and concentration of Formula A provides a previously undiscovered antibiofilm effect. This novel effect does not rely on inhibiting the growth of microorganisms on the coating, but still prevents biofilm formation on the coated surface, which is particularly advantageous from a regulatory perspective (see Example 8). Coating preparation and characterization are shown in Examples 1-3. Example 4 demonstrates that the coated surface is not cytotoxic. The amount of Formula A according to the present invention is significantly lower than that in previous techniques, as shown in Example 5 and Reference Example 6. The dose response of antibiofilm activity (versus growth inhibition) is further demonstrated in Example 7.

[0012] The present invention particularly relates to the following items: The subject matter disclosed in the following items should be considered as disclosed as if it were disclosed in the claims.

[0013] 1. A medical device comprising a coating, the coating comprising: Polymer Z attached to a medical device and part of formula A [ka] wherein X is an amino group covalently bonded to Z or capable of ionically bonding to a free carboxyl group. Including, The moiety according to formula A is 0.5-30 nmol / cm in the coating. 2 is present in an amount of Polymer Z is 1 to 30 μmol / cm 2 containing free carboxyl groups in an amount of Medical equipment.

[0014] 2. The coating has a structure of Formula I: [ka] [wherein the circle represents the surface of the medical device] The medical device according to any one of the preceding items, comprising:

[0015] 3. The coating is a compound in which X is a primary amino group -NH2, a secondary amino group -NRH, or a tertiary amino group -NR2, and each R is C 1~4 The medical device of any one of the preceding items, comprising a moiety of formula A independently selected from alkyl.

[0016] 4. Coating structure: [ka] The medical device according to any one of the preceding items, comprising:

[0017] 5. The medical device of any one of the preceding items, wherein at least 20 mol.% of the moiety according to formula A is attached to Z via a covalent bond.

[0018] 6. The medical device of any one of the preceding items, wherein at least 20 mol.% of the moiety according to formula A is ionically associated with the coating.

[0019] 7. The moiety according to formula A is 0.5-15 nmol / cm in the coating. 2 The medical device according to any one of the preceding items, wherein the amount of

[0020] 8. Polymer Z is 3-7 μmol / cm 2 The medical device of any one of the preceding items, comprising free carboxyl groups in an amount of

[0021] 9. Coating 2-(pyridyldithio)ethylamine (PDEA) covalently bound to a polymer that is covalently bonded to a medical device Including, PDEA is present in an amount of 0.001 wt% to 1 wt% based on the total weight of the coating, A medical device according to any of the preceding items.

[0022] 10. The medical device according to item 9, wherein the PDEA is present in an amount of 0.01 wt% to 1 wt% based on the total weight of the coating.

[0023] 11. The medical device of any one of the preceding items, wherein polymer Z is covalently attached to the medical device.

[0024] 12. The medical device of any one of the preceding items, wherein the coating comprises a polymer selected from the group consisting of polyacrylic acid, polymethacrylic acid, poly-4-vinylbenzoic acid, polyitaconic acid, any combination thereof, and esters or amides thereof.

[0025] 13. The medical device of any one of the preceding items, wherein polymer Z comprises an acrylate polymer or an acrylic polymer, such as polyacrylic acid.

[0026] 14. A medical device according to any one of the preceding items, wherein polymer Z of the coating is grafted from the medical device.

[0027] 15. The medical device of any one of the preceding items, wherein the coating contains water, thereby forming a hydrogel.

[0028] 16. The medical device of any one of the preceding items, selected from the group consisting of a catheter, an implant, a tracheal tube, a stent, a ventilator, a wound dressing, a face mask, a nasal prong, a hearing aid, and a syringe.

[0029] 17. The medical device according to any one of the preceding items, selected from the group consisting of a central venous catheter, a ureteral stent, a wound dressing, a ventilator, a face mask, a nasal prong and an implant.

[0030] 18. The medical device of any one of the preceding items, which is a catheter, such as a Foley catheter, or an endotracheal tube.

[0031] 19. Polymers, such as thermoplastic and / or thermosetting polymers, and / or metal The medical device according to any one of the preceding items, comprising:

[0032] 20. The polymer comprises one or more of silicone, polyethylene, polypropylene, polyurethane, polyvinyl chloride, polycaprolactone, polycarbonate, rubber such as latex rubber, polyetheretherketone (PEEK); and / or the metal comprises one or more of a steel such as stainless steel, an alloy such as a cobalt-based alloy or nitinol, or titanium; Item 19. The medical device according to item 19.

[0033] 21. A medical device according to any one of the preceding items, wherein the coating is present on at least a portion of the interior and / or exterior surface of the medical device.

[0034] 22. The medical device of any one of the preceding paragraphs, which is free or substantially free of biofilm after 4 days or more, e.g., 30 days, of clinical use.

[0035] 23. A coating for a medical device as defined in any one of the preceding paragraphs.

[0036] 24. Use of a coating to reduce the formation and / or growth of biofilms on a surface of a medical device, the coating comprising: Polymer Z attached to a medical device and part of formula A [ka] wherein X is an amino group covalently bonded to Z or capable of ionically bonding to a free carboxyl group. Including, Polymer Z is capable of forming a hydrogel when in contact with a fluid containing water, use.

[0037] 25. The use according to item 24, wherein polymer Z is covalently attached to the medical device.

[0038] 26. Use according to item 24 or 25, wherein Z contains a free carboxyl group.

[0039] 27. Z is 1-30 μmol / cm 2 27. Use according to item 26, containing free carboxyl groups in an amount of

[0040] 28. Z is 3-20 μmol / cm 2 28. Use according to item 27, containing free carboxyl groups in an amount of

[0041] 29. The coating has a structure of Formula I: [ka] [where the circle represents the surface of the medical device] 29. The use according to any one of items 24 to 28, comprising:

[0042] 30. A coating comprising: X is a primary amino group -NH2, a secondary amino group -NRH, or a tertiary amino group -NR2; and each R is C 1~4 30. The use according to any of items 24 to 29, comprising a moiety of formula A independently selected from alkyl.

[0043] 31. Coating structure: [ka] 31. The use according to any one of items 24 to 30, comprising:

[0044] 32. Use according to any of items 24 to 31, wherein at least 20 mol.% of moieties according to formula A are attached to Z via a covalent bond.

[0045] 33. Use according to any of items 24 to 32, wherein at least 20 mol.% of moieties according to formula A are ionically associated with the coating.

[0046] 34. The fraction following formula A is 0.5-30 nmol / cm 2 34. Use according to any one of items 24 to 33, wherein the coating is present in an amount of

[0047] 35. The fraction following formula A is 0.5-15 nmol / cm 2 35. Use according to item 34, wherein the coating is present in an amount of

[0048] 36. Use according to any one of items 24 to 35, wherein the coating comprises a polymer selected from the group consisting of polyacrylic acid, polymethacrylic acid, poly-4-vinylbenzoic acid, polyitaconic acid, any combination thereof or esters or amides thereof.

[0049] 37. Use according to any one of items 24 to 36, wherein polymer Z comprises an acrylate polymer or an acrylic polymer, such as polyacrylic acid.

[0050] 38. Use according to any one of items 24 to 37, wherein the polymer Z of the coating is grafted from the medical device.

[0051] 39. Use according to any one of items 24 to 38, wherein the coating is as defined in any one of items 1 to 22.

[0052] 40. Use according to any one of items 24 to 39, wherein the alleviation is not accompanied by a bactericidal or bacteriostatic effect.

[0053] The arrangement of this disclosure into sections with headings and subheadings is merely to improve readability and should not be construed as limiting in any way, and in particular this division does not in any way prevent or restrict features under different headings and subheadings from being combined with one another. All references are incorporated herein by reference. [Brief explanation of the drawings]

[0054] [Figure 1] FIG. 1 shows the optical density (OD) measured at a wavelength of 595 nm for three catheter samples described in Example 2. OD correlates with the EPS matrix and the number of embedded bacteria, demonstrating that the coating of the present invention results in near complete inhibition of biofilm formation. [Figure 2] FIG. 1 shows the measured cumulative amount of 2-mercaptopyridine, and therefore the amount of PDEA, in the coatings measured in Example 5. [Figure 3] FIG. 1 shows the measured cumulative amount of 2-mercaptopyridine, and therefore the amount of PDEA, in the coating measured in Reference Example 6. [Figure 4] Using cycle 1 as a baseline, a zoomed-in view from cycle 2 onwards is shown in Figure 3. This shows the amount of PDEA remaining in the previous generation product after coupling with the cysteine ​​moiety (corresponding to the situation after one cycle of coupling). [Figure 5] Figure 1 shows the number of attached bacteria present on the surface of the sample, expressed in log CFU / cm2. It can be seen that low concentrations have no effect on the number of bacteria. [Figure 6] Figure 1 shows the number of bacteria in artificial urine medium (AUM) expressed in log CFU / ml, showing that low concentrations have no effect on bacterial counts. [Figure 7] Figure 1 shows the number of bacteria in artificial urine medium (AUM) and on the catheter surface, expressed in log CFU / ml. It can be seen that the coating has no effect on the number of bacteria in the solution, indicating that it has no bactericidal or bacteriostatic effect (CytaCoat refers to the coating of the present invention). [Figure 8]Quantification of biofilm on catheter surfaces. A coating of the present invention (designated "CytaCoat") almost completely prevents biofilm formation during the first week (right panel). When bacteria from week 1 are placed in contact with a control catheter, they are still able to form biofilm during week 2 (left panel), indicating that exposure to the surface of the present invention did not alter their properties. [Figure 9] FIG. 1 shows the experimental setup for Example 8. DETAILED DESCRIPTION OF THE INVENTION

[0055] Medical devices with coatings In a first aspect, the present invention provides a medical device comprising a coating, the coating comprising: Polymer Z attached (preferably covalently attached) to the medical device and part of formula A [ka] wherein X is an amino group covalently bonded to Z or capable of ionically bonding to a free carboxyl group. Including, The moiety according to formula A is approximately 0.5-30 nmol / cm in the coating. 2 is present in an amount of Polymer Z is approximately 1 to 30 μmol / cm 2 containing free carboxyl groups in an amount of Provide medical equipment.

[0056] The present invention relates to a medical device comprising a coating, the coating comprising a polymer Z attached (preferably covalently attached) to the medical device, and a disulfide compound of formula (I) as defined in WO 2023 / 012305 (which is incorporated herein in its entirety) covalently attached or non-covalently associated with the polymer, the polymer Z preferably being present in a concentration of about 1 to 30 μmol / cm 2The present invention also relates to medical devices containing free carboxyl groups in an amount of 10 to 12. The disulfide compounds disclosed in claims 10, 11 or 12 of WO 2023 / 012305 are particularly preferred.

[0057] X can be a primary amino group -NH2, a secondary amino group -NRH, or a tertiary amino group -NR2, and each R is C 1~4 C is independently selected from alkyl. 1~4 Examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. In some embodiments, X is a primary amino group -NH2.

[0058] X can be covalently bonded to Z at a terminal functional group and / or to a functional group along the polymer backbone. An example of such a functional group is carboxy. Alternatively, X can be ionically bonded to a free group at the end or to a free group (e.g., a carboxyl group) along the polymer backbone. For example, if X is a primary amino group -NH2 and the polymer contains -COOH groups, these groups will have opposite charges (-NH3 + , -COO - ) and can therefore associate ionically. Similarly, the secondary amino group -NRH can be expressed in its protonated form -NRH2 + -COO - The tertiary amino group -NR2 can be ionically bonded to its protonated form -NR2H + -COO - can be ionically bonded to

[0059] The coating has a structure of Formula I: [ka] [wherein the circle represents the surface of the medical device] The coating may additionally or alternatively comprise a moiety of formula A, as defined above, where X is an amino group. In some embodiments, the coating has the structure: [ka] Includes:

[0060] At least about 1 mol.% or, in order of increasing preference, at least about 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 mol.% such as 2-100, 5-100, 10-100, 30-100, 50-100, 80-100, 2-90, 5-90, 10-90, 20-90, 30-90, 50-90, 80-90, 2-70, 5-70, 10-70, 30-70, 50-70, 20-80, 30-90, 40-80 mol.% of the moiety according to formula A may be attached to Z via a covalent bond. At least 1 mol.% or, in order of increasing preference, at least about 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 mol.% e.g. 2-100, 5-100, 10-100, 30-100, 50-100, 80-100, 2-90, 5-90, 10-90, 20-90, 30-90, 50-90, 80-90, 2-70, 5-70, 10-70, 30-70, 50-70, 20-80, 30-90, 40-80 mol.% of moieties according to Formula A may be ionically associated with the coating. For the avoidance of doubt, it is possible for both covalently bound and ionically associated moieties to be present simultaneously.

[0061] The moiety according to formula A is approximately 0.5-15 nmol / cm in the coating. 2 In certain embodiments, the moiety according to Formula A may be present in an amount of about 0.5 to 12 nmol / cm 2 , about 0.5~10nmol / cm 2 , about 0.5~9nmol / cm 2 , about 0.5~8nmol / cm 2 , about 0.5~7nmol / cm 2 , about 0.5~6nmol / cm2 , about 0.5~5nmol / cm 2 , about 0.5~4nmol / cm 2 , about 0.5~3nmol / cm 2 , about 0.5~2nmol / cm 2 , about 0.5~1nmol / cm 2 , about 1~12nmol / cm 2 , about 1~10nmol / cm 2 , about 1~9nmol / cm 2 , about 1~8nmol / cm 2 , about 1~7nmol / cm 2 , about 1~6nmol / cm 2 , about 1~5nmol / cm 2 , about 1~4nmol / cm 2 , about 1~3nmol / cm 2 , about 1~2nmol / cm 2 , or approximately 1 nmol / cm 2 may be present in an amount of

[0062] The polymer Z preferably has a concentration of at least 3 μmol / cm 2 , for example, about 3 to 20 μmol / cm 2 In certain embodiments, the surface concentration of carboxylic acid groups may be about 1-20 μmol / cm 2 , about 1~15μmol / cm 2 , about 1~10μmol / cm 2 , about 1~7μmol / cm 2 or approximately 1-5 μmol / cm 2 It can be about 3 to 7 μmol / cm 2 A surface concentration of carboxylic acid groups of which is most preferred.

[0063] In one set of alternative embodiments of the first aspect, the invention is a medical device comprising a coating, the coating comprising: 2-(pyridyldithio)ethylamine (PDEA) covalently bound to a polymer that is covalently bonded to a medical device Including, PDEA is present in an amount of 0.001 wt% to 35 wt% based on the total weight of the coating, Provide medical equipment.

[0064] PDEA can be present in an amount of about 0.001 wt% to about 35 wt%, e.g., about 0.001 wt% to about 30 wt%, about 0.001 wt% to about 25 wt%, about 0.001 wt% to about 20 wt%, about 0.001 wt% to about 15 wt%, about 0.001 wt% to about 10 wt%, about 0.001 wt% to about 5 wt%, 0.001 wt% to about 3 wt%, 0.001 wt% to about 2 wt%, about 0.001 wt% to about 1 wt%, or 0.001 wt% to about 0.5 wt%, based on the total weight of the coating. For example, PDEA can be present in an amount of about 0.001 wt% to about 2 wt%, or about 0.001 wt% to about 1 wt%, based on the total weight of the coating. In a further example, the amount of PDEA can be about 0.5 wt% to about 2 wt%, e.g., about 0.5 wt% to about 1.5 wt%, based on the total weight of the coating. In a further example, the amount of PDEA can be about 0.001 wt% to about 1 wt%, 0.01 wt% to 1 wt%, 0.05 wt% to 1 wt%, 0.08 wt% to 1 wt%, or 0.1 wt% to 1 wt%, based on the total weight of the coating.

[0065] It will be understood that the compound 2-(pyridyldithio)ethylamine may be abbreviated as PDEA. Furthermore, the compound structure of 2-(pyridyldithio)ethylamine (PDEA) may be represented as shown below. In this document, the chemical name 2-(pyridyldithio)ethylamine may be used interchangeably with 2-(pyridin-2-yl)disulfanamine, 2-(pyridin-2-yldisulfanyl)ethanamine, 2-(2-pyridyldithio)ethylamine and / or PDEA. [ka]

[0066] In certain contexts herein, the term "PDEA" may also be used more broadly to refer to chemical structures containing a moiety according to Formula A. PDEA is known to react with thiol-containing nucleophiles, such as cysteine, to replace the sulfur attached to the pyridine ring of PDEA with the sulfur from the thiol group of the nucleophile, cleaving the disulfide bond of PDEA. As a result, a compound containing -S-(CH)-NH derived from PDEA is formed along with pyridine-2-thiol. This is illustrated in Scheme 1 below, where the nucleophile R-SH is reacted with PDEA. [ka]

[0067] Similarly, the reaction of thiol-containing nucleophiles with PDEA has been used in applications where, for example, the amino group of PDEA is covalently bonded to a carbonyl group of a polymer, and the polymer is then covalently or otherwise bonded to the surface of a substrate such as a medical device. This is illustrated in Scheme 2, where the circle represents a surface such as a medical device surface, Z represents the polymer, and the carbonyl group is part of the polymer. [ka]

[0068] For example, WO 2006 / 101438 A1 describes such reactions using cysteine ​​or cysteine ​​analogues as thiol-containing nucleophiles that result in surfaces with excellent antimicrobial properties.

[0069] Preventing or reducing biofilm In this context, the term biofilm refers to complex three-dimensional structures on surfaces (e.g., medical devices) formed by a community of microorganisms (bacteria, fungi, or protists) embedded in self-produced extracellular polymeric substances (EPS). Biofilm formation protects microorganisms, making them difficult to eradicate and potentially significantly affecting their pathogenicity. Biofilm formation is a multi-step process involving attachment, EPS production, and biofilm maturation.

[0070] The terms prevention and reduction in the context of biofilm formation and / or growth refer to any degree of reduction in biofilm formation, including slight, substantial, or large reductions in biofilm formation, as well as complete prevention. Preferably, the degree of reduction is at least slight. The degree of reduction can be at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100%. For example, the degree of reduction can be about 10% to about 100%, e.g., about 20% to about 100%, about 40% to about 100%, about 60% to about 100%, about 80% to about 100%, about 20% to about 40%, about 20% to about 60%, about 20% to about 80%, about 20% to about 100%, about 40% to about 60%, about 40% to about 80%, about 40% to about 100%, about 60% to about 80%, about 60% to about 100%, or about 80% to about 100%. "Anti-biofilm effect" refers to a reduction in biofilm formation and / or growth.

[0071] The present invention is based on the unexpected discovery that reduction of biofilm formation and / or growth on a surface, such as a medical device surface, can be achieved by covalently attaching PDEA or other moieties according to Formula A to a polymer that is covalently or otherwise attached to the surface of the medical device. Alternatively, PDEA or other compounds containing a moiety according to Formula A can be non-covalently (e.g., ionically) associated with the polymer.

[0072] Therefore, reaction of PDEA or a moiety according to Formula A with additional chemical compounds is not required to achieve mitigation of biofilm formation and / or growth. A possible chemical structure of such a functionalized surface is shown by Formula I, where circle and Z have the meanings described herein and the carbonyl group is part of the polymer. [ka]

[0073] In particular, it has been found that the amount of the moiety of Formula A can be low, such as provided in the amounts described herein, and still allow for the reduction of biofilm formation and / or growth on the surface of a medical device. At low amounts, the coating has neither a bacteriostatic nor a bactericidal effect, yet the anti-biofilm effect prevents fouling of the medical device.

[0074] Of course, using small amounts of PDEA or other structures containing Formula A as the sole anti-biofilm agent is a significant advantage because it simplifies manufacturing and / or minimizes chemical use. Additionally, using small amounts of a structure according to Formula A covalently or non-covalently attached to a polymer minimizes the risk of toxicity. As shown herein, testing of the coating did not demonstrate cytotoxicity. Thus, there is no or minimal release of PDEA (or other compounds containing a structure according to Formula A) in therapeutically effective amounts. This, in turn, simplifies the regulatory process for medical devices bearing the coating.

[0075] It will be understood that biofilms include microorganisms such as bacteria. The bacteria can be gram-positive and / or gram-negative bacteria. For example, the bacteria can include one or more of the following bacteria: Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus viridans, Enterococcus faecium, Enterococcus faecium, Streptococcus spp, E. coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, and cinetobacter baumannii. In one example, the Gram-positive bacteria can be one or more of the following: Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus viridans, Enterococcus faecium, Enterococcus faecium, Streptococcus spp. In a further example, the Gram-negative bacteria can be one or more of the following: E. coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa. Additionally or alternatively, the biofilm may include a fungus such as Candida Albicans.

[0076] It will be understood that, as used herein, mitigating biofilm formation and / or growth can involve minimizing, reducing, and / or preventing biofilm formation and / or growth. Without wishing to be bound by any particular theory, it is believed that some bacteria may still adhere to surfaces but not aggregate into biofilms, thereby making them more susceptible to treatment with antimicrobial agents such as antibiotics, or simply allowing them to be washed or flushed away. It is also known that microorganisms without biofilm / EPS are less likely to cause infectious diseases (i.e., less aggressive as pathogens). The lack of biofilm also makes them more susceptible to the body's immune system or more susceptible to drug treatment.

[0077] Coating polymer The polymer (which may be covalently attached to the moiety of Formula A) becomes a coating on the surface. In the presence of water, the coating can form a hydrogel. Thus, there is provided a coating described herein that includes or consists of a hydrogel. As used herein, a hydrogel is a cross-linked hydrophilic polymer that is insoluble in water.

[0078] It will be appreciated that the polymer of the coating may be bonded, such as by a covalent bond, to the surface of the medical device. This is believed to allow the coating to remain in place and increase resistance to abrasion. Advantageously, the coated medical devices of the present invention have been found to be abrasion resistant.

[0079] Functionalization of medical device surfaces involves polymerization of a monomer prior to reaction with PDEA or other compounds containing the structure of Formula A. The monomer may contain one or more functional groups, such as carboxyl groups, amino groups, halogens, etc.

[0080] The monomer may be selected from the group consisting of acrylic acid, methacrylic acid, 4-vinylbenzoic acid, itaconic acid, vinylpyrrolidone, any combination thereof, and esters or amides thereof. For example, the monomer may comprise or consist of an acrylate monomer or an acrylic monomer, such as acrylic acid. Thus, the polymer of the coating may be or comprise polyacrylic acid, polymethacrylic acid, poly-4-vinylbenzoic acid, polyitaconic acid, any combination thereof, and esters or amides thereof. The coating may further comprise polyvinylpyrrolidone. In particular, the polymer may comprise an acrylate polymer or an acrylic polymer, such as polyacrylic acid. For example, the polymer may comprise or consist of polyacrylic acid. For polymers containing carboxylic acid groups, the surface concentration of carboxylic acid may be greater than or equal to about 1 μmol / cm. 2 ~about 30μmol / cm 2 , for example, about 5 μmol / cm 2 ~about 25μmol / cm 2 , about 5μmol / cm 2 ~about 20μmol / cm 2 , about 5μmol / cm 2 ~about 10μmol / cm 2 , about 6μmol / cm 2 ~about 9 μmol / cm 2 For example, the surface concentration of carboxylic acid can range from about 3 μmol / cm 2 ~about 8μmol / cm 2 , about 4μmol / cm 2 ~about 7μmol / cm 2 or approximately 5 μmol / cm 2 ~about 6 μmol / cm 2 In certain embodiments, the surface concentration of carboxylic acid groups can range from about 1 to 20 μmol / cm 2 , about 1~15μmol / cm 2 , about 1~10μmol / cm 2 , about 1~7μmol / cm 2 or approximately 1-5 μmol / cm 2 It can be about 3 to 7 μmol / cm 2A surface concentration of carboxylic acid groups of which is most preferred.

[0081] The polymer of the coating can be grafted from the medical device. In this way, polymerization is initiated and propagated from the surface, with the resulting polymer being covalently attached to the surface. Additionally or alternatively, a preformed polymer can be covalently attached to the surface of the medical device by grafting the polymer onto the surface.

[0082] Appropriate medical equipment The medical devices described herein may be selected from the group consisting of catheters, implants, tracheal tubes, stents, ventilators, wound dressings, face masks, nasal prongs, hearing aids, and syringes. In a further example, the medical device may be selected from the group consisting of central venous catheters, ureteral stents, wound dressings, ventilators, face masks, nasal prongs, and implants. In a further example, the medical device may be a catheter, such as a Foley catheter, or an endotracheal tube.

[0083] The medical devices described herein may comprise or consist of a polymer, such as a thermoplastic and / or thermosetting polymer. For example, the polymer may comprise or consist of one or more of the following: silicone, polyethylene, polypropylene (PP), polyurethane, polyvinyl chloride (PVC), polycaprolactone, polycarbonate, rubber such as latex rubber, polyetheretherketone (PEEK). In one example, the polymer may comprise one or more of the following: silicone, polyvinyl chloride (PVC), polypropylene (PP). In a further example, the polymer may comprise or consist of silicone. In a further example, the polymer may comprise or consist of polyvinyl chloride (PVC). In a further example, the polymer may comprise or consist of polypropylene (PP).

[0084] Additionally or alternatively, the medical device may comprise or consist of a metal. For example, the metal may comprise one or more of the following: a steel, such as stainless steel, an alloy, such as a cobalt-based alloy or nitinol, or titanium. As used herein, nitinol is an alloy comprising or consisting of nickel and titanium.

[0085] The coating of the medical device can be present on at least a portion of the interior and / or exterior surface of the medical device.

[0086] For example, it has been found that the surfaces of the medical devices described herein can remain free or substantially free of biofilm after 4 or more days of clinical use, e.g., the medical devices can remain free of biofilm after 7, 14, 21, or 30 days of clinical use.

[0087] Coatings can be applied to medical devices using methods known in the art. For example, the methods described in WO 2006 / 101438 A1 can be used to prepare the coated medical devices described herein. It will be understood, however, that the bound PDEA or other structure containing the structure of Formula A is preferably not further reacted. For example, the medical device surface can be functionalized by polymerizing a monomer, such as the monomers described herein, from the device surface using irradiation, such as electron beam irradiation, gamma irradiation, UV irradiation, or gas plasma, in the presence of a photoinitiator. The formed polymer can then be further reacted with PDEA or other moieties of Formula A in the presence of a coupling agent, such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysulfosuccinimide (sulfo-NHS) or a salt thereof. Alternatively, instead of using a coupling agent, the carboxylic acid group can be converted to an acyl chloride or anhydride. This is illustrated in Scheme 3, where the circle represents a surface, such as a medical device surface, Z represents the polymer, and the carboxylic acid group is part of the polymer. [ka]

[0088] Coatings for medical devices In a second aspect, there is provided a coating for a medical device as defined in the first aspect. In particular, there is provided a coating comprising a polymer of formula II: [ka]

[0089] In the compound of Formula II, Z can be a polymer described herein. For example, Z can be an acrylate polymer or an acrylic polymer, such as polyacrylic acid. Additionally, the carbonyl group in the polymer of Formula I can be part of the polymer Z. The polymer of Formula II can be attached, such as by a covalent bond, to a surface, such as the surface of a medical device. For example, the polymer of Formula II can be attached to the surface via a covalent bond from Z to the surface. The coating can further include water, thereby providing the coating as a hydrogel. The coating can be applied to the surface of a medical device described herein.

[0090] Use of coatings to prevent biofilm formation on medical device surfaces In a third aspect, the present invention provides the use of a coating to reduce the formation and / or growth of biofilms on a surface of a medical device.

[0091] The coating is Polymer Z attached (preferably covalently attached) to the medical device and part of formula A [ka] wherein X is an amino group covalently bonded to Z or capable of ionically bonding to a free carboxyl group. and Polymer Z can form a hydrogel when in contact with a water-containing fluid, such as water, an aqueous buffer solution, saline, or a bodily fluid (including, but not limited to, urine, plasma, blood, wound fluid, saliva, and cerebrospinal fluid).

[0092] Polymer Z may contain free carboxyl groups. Polymer Z preferably has a concentration of about 1 to 30 μmol / cm 2 In certain embodiments, the surface concentration of carboxylic acid groups may be at least about 3 μmol / cm 2 , about 1~20μmol / cm 2 , about 3~20μmol / cm 2 , about 1~15μmol / cm 2 , about 1~10μmol / cm 2 , about 1~7μmol / cm 2 or approximately 1-5 μmol / cm 2 It can be about 3 to 7 μmol / cm 2 A surface concentration of carboxylic acid groups of which is most preferred.

[0093] X can be a primary amino group -NH2, a secondary amino group -NRH, or a tertiary amino group -NR2, and each R is C 1~4 C is independently selected from alkyl. 1~4 Examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. In some embodiments, X is a primary amino group -NH2.

[0094] X may be covalently bonded to Z at a terminal functional group and / or to a functional group along the polymer backbone. An example of such a functional group is carboxy. Alternatively, X may be ionically bonded to a terminal free carboxyl group or a free carboxyl group along the polymer backbone, as discussed in the first embodiment.

[0095] The coating has a structure of Formula I: [ka] [wherein the circle represents the surface of the medical device] The coating may additionally or alternatively comprise a moiety of formula A, where X is an amino group. Thus, the coating may have the structure: [ka] may include:

[0096] At least about 1 mol.%, or in order of increasing preference at least about 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 mol.%, for example 2-100, 5-100, 10-100, 30-100, 50-100, 80-100, 2-90, 5-90, 10-90, 20-90, 30-90, 50-90, 80-90, 2-70, 5-70, 10-70, 30-70, 50-70, 20-80, 30-90, 40-80 mol.% of the moiety according to formula A may be attached to Z via a covalent linker. At least about 1 mol.% or, in order of increasing preference, at least about 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 mol.% e.g. 2-100, 5-100, 10-100, 30-100, 50-100, 80-100, 2-90, 5-90, 10-90, 20-90, 30-90, 50-90, 80-90, 2-70, 5-70, 10-70, 30-70, 50-70, 20-80, 30-90, 40-80 mol.% of the moiety according to Formula A may be non-covalently associated with the coating. It is possible for both covalently bound and non-covalently associated moieties to be present simultaneously.

[0097] The moiety according to formula A is approximately 0.5-30 nmol / cm in the coating. 2 In certain embodiments, the moiety according to Formula A may be present in an amount of about 0.5 to 15 nmol / cm 2 , 0.5-12 nmol / cm 2 , about 0.5~10nmol / cm 2 , about 0.5~9nmol / cm 2, about 0.5~8nmol / cm 2 , about 0.5~7nmol / cm 2 , about 0.5~6nmol / cm 2 , about 0.5~5nmol / cm 2 , about 0.5~4nmol / cm 2 , about 0.5~3nmol / cm 2 , about 0.5~2nmol / cm 2 , about 0.5~1nmol / cm 2 , about 1~12nmol / cm 2 , about 1~10nmol / cm 2 , about 1~9nmol / cm 2 , about 1~8nmol / cm 2 , about 1~7nmol / cm 2 , about 1~6nmol / cm 2 , about 1~5nmol / cm 2 , about 1~4nmol / cm 2 , about 1~3nmol / cm 2 , about 1~2nmol / cm 2 , or approximately 1 nmol / cm 2 may be present in an amount of

[0098] The coating may comprise a polymer Z selected from the group consisting of polyacrylic acid, polymethacrylic acid, poly-4-vinylbenzoic acid, polyitaconic acid, any combination thereof, and esters or amides thereof. The coating may further comprise polyvinylpyrrolidone. Polymer Z may comprise an acrylate polymer or an acrylic polymer, such as polyacrylic acid. Polymer Z of the coating may be grafted from the medical device.

[0099] The coating may be as defined in the first or second aspect.

[0100] Preferably, the reduction in biofilm formation and / or growth is not accompanied by a bactericidal or bacteriostatic effect.

[0101] Itemized embodiments In certain embodiments, the present invention relates to the following items:

[0102] 1. A medical device comprising a coating, the coating comprising: 2-(pyridyldithio)ethylamine (PDEA) covalently bound to a polymer that is covalently bonded to a medical device Including, PDEA is present in an amount of 0.001 wt% to 35 wt% based on the total weight of the coating, Medical equipment.

[0103] 2. The medical device according to item 1, wherein the PDEA is present in an amount of 0.001 wt% to 1 wt% based on the total weight of the coating.

[0104] 3. The medical device of item 1 or item 2, wherein the polymer is selected from the group consisting of polyacrylic acid, polymethacrylic acid, poly-4-vinylbenzoic acid, polyitaconic acid, polyvinylpyrrolidone, any combination of the foregoing, and esters or amides thereof.

[0105] 4. The medical device of any one of the preceding items, wherein the polymer comprises an acrylate polymer or an acrylic polymer, such as polyacrylic acid.

[0106] 5. The medical device of any one of the preceding items, wherein the polymer of the coating is grafted from the medical device.

[0107] 6. The medical device of any one of the preceding items, wherein the coating contains water, thereby forming a hydrogel.

[0108] 7. The medical device of any one of the preceding items, selected from the group consisting of a catheter, an implant, a tracheal tube, a stent, a ventilator, a wound dressing, a face mask, a nasal prong, a hearing aid, and a syringe.

[0109] 8. The medical device according to any one of the preceding items, selected from the group consisting of a central venous catheter, a ureteral stent, a wound dressing, a ventilator, a face mask, a nasal prong, and an implant.

[0110] 9. The medical device of any one of the preceding items, which is a catheter, such as a Foley catheter, or an endotracheal tube.

[0111] 10. Polymers, such as thermoplastic and / or thermosetting polymers, and / or metal The medical device according to any one of the preceding items, comprising:

[0112] 11. The polymer comprises one or more of silicone, polyethylene, polypropylene, polyurethane, polyvinyl chloride, polycaprolactone, polycarbonate, rubber such as latex rubber, polyetheretherketone (PEEK); and / or the metal comprises one or more of a steel such as stainless steel, an alloy such as a cobalt-based alloy or nitinol, or titanium; Item 10. The medical device according to item 10.

[0113] 12. The medical device of any one of the preceding items, wherein the coating is present on at least a portion of the interior and / or exterior surface of the medical device.

[0114] 13. The medical device of any one of the preceding paragraphs, which is free or substantially free of biofilm after 4 days or more, e.g., 30 days, of clinical use.

[0115] 14. A coating for a medical device as defined in any one of the preceding paragraphs.

[0116] 15. Use of a coating as defined in any one of the preceding paragraphs to reduce the formation and / or growth of biofilms on the surface of a medical device.

[0117] The term "comprising" is to be interpreted as including but not limited to.

[0118] Numerical values ​​in the specification and claims of this application should be understood to include numerical values ​​that would be the same if reduced to the correct number of significant digits, and numerical values ​​that differ from the stated value by less than the experimental error of conventional measurement techniques of the type described in this application for determining the value.

[0119] All ranges disclosed herein are inclusive of the recited endpoints and are independently combinable (e.g., the range "from 2 to 10" includes the endpoints 2 and 10, and all intermediate values).

[0120] The term "about" can be used to include any numerical value that can vary without changing the basic function of that value. When used in conjunction with a range, "about" also discloses the range defined by the absolute values ​​of the two endpoints, for example, "about 2 to about 4" discloses the range "2 to 4." The term "about" can refer to ±10% of the indicated number.

[0121] The present invention is further described with reference to the following examples, which are not intended to limit the scope of the invention. [Example]

[0122] As used herein, the chemical name "2-(pyridin-2-yldisulfanyl)ethanamine" and chemical structure of the compound of Formula I were generated using the program ChemDraw Ultra version 12.0.2.1076. In the event of a conflict between the drawing and the chemical name, the drawing shall be considered correct.

[0123] Abbreviation AUM artificial urine medium aq.water-based BP Benzophenone cfu colony forming unit cm 2 square centimeter DI Deionization g grams NHS N-hydroxysuccinimide; CAS number: 6066-82-6T EDC 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide; CAS number: 25952-53-8 EPS extracellular polymeric substance ISO International Organization for Standardization L liters L-Cysteine ​​CAS number: 52-90-4 mg milligram μmol micromol min. ml milliliter M molar concentration MQ Milli-Q nm nanometer nmol nanomol OD optical density PAA Polyacrylic acid; CAS number: 9003-01-4 PBS Phosphate Buffered Saline PDEA 2-(pyridyldithio)ethylamine; CAS number: 106139-15-5 Sulfo-NHS N-hydroxysulfosuccinimide UV ultraviolet wt% weight percent

[0124] The AUM used herein had the same composition as that described in Letters in Applied Microbiology 1997, 24, 203-206.

[0125] Example 1: Preparation of anti-biofilm silicone Foley catheters A coating process protocol for applying an anti-biofilm coating onto a catheter is described below, and it will be understood that this protocol can be applied to a single catheter.

[0126] PAA hydrogel 1. The catheter was washed in ethanol for 1 minute. 2. The catheter was allowed to dry at ambient temperature for 10 minutes. 3. The catheter was immersed in an ethanol solution of the photoinitiator BP (5%) and then irradiated with UV light for 30 seconds. 4. The catheter was washed in ethanol for 30 seconds to remove excess BP. 5. A 10% solution of acrylic acid in water was prepared: for each liter of acrylic acid solution: 900 ml DI water, 100 ml acrylic acid, Mohr's salt (1.65 g), and Cu(NO3)2 (1.65 g). The solution was degassed by adding a stir bar to the flask, placing the flask on a stir plate, and stirring under vacuum until no more bubbles were visible. Mohr's salt, as used herein, refers to ammonium iron(II) sulfate. 6. The catheter was immersed in the prepared aqueous monomer solution and then irradiated with UV light for approximately 4 minutes. 7. During sonication, the catheter was immersed in DI water for 10 minutes twice. 8. During the sonication, the catheter was immersed in ethanol for 60 minutes. 9. The catheter was allowed to dry for 30 minutes.

[0127] Hydrogel characterization: The surface concentration of grafted polyacrylic acid was determined by adding a known amount of NaOH (aq) to the catheter segment in a test tube and then shaking the test tube for 4 to 24 hours. The remaining NaOH (aq) was titrated with HCl (aq), and the HCl consumed in the titration was used as the basis for calculating the number of carboxylic acid groups. The number of carboxylic acid groups was then divided by the surface area of ​​the catheter segment to obtain the surface concentration of carboxylic acid groups on the polyacrylic acid. 1 μmol / cm 2 ~30 μmol / cm 2 It was found that a surface concentration of carboxylic acid groups of 0.01% could be achieved.

[0128] PDEA Coupling Process 1. Prepare an EDC / NHS solution. The ratio is 4:1. 0.1M / 0.025M. 2. The catheter was immersed in the EDC / NHS solution for approximately 10 minutes. 3. The catheter was immersed in DI water for 15 seconds three times. 4. Prepare PDEA solutions in borate buffer solution: For each liter of borate buffer solution: Mix 6.18 g of H3BO3 in 1 L of DI water. Adjust the pH to 8.5 with 1 M NaOH. Add 21 mg of PDEA to each liter of borate buffer and mix in a glass flask. 5. The catheter was immersed in the PDEA solution for 10 minutes. 6. The catheter was immersed in DI water for 60 seconds three times. 7. The catheter was immersed in 1M phosphate buffer, pH 7.4, for 6 hours. 8. The catheter was immersed in DI water for 1 minute three times. 9. The catheter was allowed to dry at ambient temperature for at least 6 hours.

[0129] Rationale for the amount of bound PDEA In one example, the estimated loading of PDEA was 0.3 wt% based on the total weight of the coating. The rationale for this estimation is based on the fact that the ratio of EDC / NHS+PDEA to polyacrylic acid was 1:1000. In other words, one in every 1000 carboxylic acid groups on the polyacrylic acid reacted with PDEA, resulting in a loading of 0.3 wt% based on the total weight of the coating, which is approximately 6 nmol / cm. 2 is equivalent to

[0130] Example 2: Testing biofilm formation and / or growth Silicone Foley catheter segments were functionalized with polyacrylic acid and PDEA as described in Example 1. The size of the catheter segments was 2 cm (16F) each.

[0131] Three uncoated catheter sections were prepared as control samples. The control samples were subjected to ethylene oxide sterilization. The control samples were designated as Sample A.

[0132] Three catheter segments with coating were not subjected to sterilization. These samples were designated Sample B.

[0133] Three catheter segments with the coating were subjected to sterilization using ethylene oxide. These samples were designated Sample C.

[0134] Catheter segments from Samples A, B, and C were exposed to Klebsiella pneumoniae bacteria as described in the protocol below.

[0135] Protocol 1. On the first day of the test, 9 One ml of freshly prepared AUM solution containing cfu / ml of Klebsiella pneumoniae (AO15200) was added to the test tube. 2. The samples were incubated at 37°C for 7 days, with the AUM solution replaced every day except on weekends. 3. On day 8, samples were fixed as follows: a) 1% glutaraldehyde in PBS for 5 minutes; b) 60% ethanol for 15 minutes; c) 80% ethanol for 15 minutes; d) dried. 4. The samples were stained with 0.04% crystal violet in water for 5 minutes, then rinsed in MQ water and dried, thus staining any bacteria or biofilm present on the sample surface. 5. The sample was transferred to adhesive tape and photographed. 6. The crystal violet from the adhesive tape strip was dissolved in 2 ml of 95% ethanol and the optical density was determined at a wavelength of 595 nm using a spectrophotometer.

[0136] With this protocol, the photographs showed clear staining for the specimens of Sample A. In contrast, the specimens of Sample B and Sample C were virtually unstained.

[0137] Furthermore, the results of spectrophotometric measurements were as shown in Table 1. [Table 1]

[0138] The average optical density of the samples in Table 1 was calculated and is illustrated in Figure 1. As shown in Table 1 and Figure 1, the Sample B and Sample C specimens had significantly less staining than the Sample A specimens.

[0139] Based on photographic and spectrophotometric measurements, it was concluded that biofilm formation and / or growth was significantly reduced in the presence of the PDEA-containing coating, as observed for Sample B and Sample C, respectively.

[0140] Example 3: Abrasion resistance test In this example, the abrasion resistance of coated catheters prepared as described in Example 1 was tested. Four different test methods were used on coated catheters or coated tubing segments of catheters (e.g., 6 cm long): (1) dry pinch test, (2) wet pinch test, gel pinch test, and (4) bend test. The test method protocols were as follows:

[0141] Dry Pinch Test This was done on coated catheters or on sections of coated tubing from catheters (eg, 6 cm long).

[0142] 1. To hold the coated catheter (or coated catheter fragment), use a dry, gloved hand to gently pinch the coated catheter between your index finger and thumb. Gently rub the surface of the coated catheter longitudinally within a 4 cm area 10 times.

[0143] 2. Subject the abraded coated catheter (or coated catheter fragment) to the dye test described herein to visually assess the coating's resistance to damage and minor abrasion compared to a coated control (no dry abrasion).

[0144] Wet Pinch Test This was done on coated catheters or on sections of coated tubing from catheters (eg, 6 cm long).

[0145] 1. To hold the coated catheter (or coated catheter fragment), use a moistened gloved hand to gently pinch the coated catheter between your index finger and thumb. Gently rub the surface of the coated catheter longitudinally within a 4 cm area 10 times.

[0146] 2. Subject the abraded coated catheter (or coated catheter fragment) to the dye test described herein to visually assess the coating's resistance to damage and minor abrasion compared to a coated control (no wet abrasion).

[0147] Gel pinch test This was done on coated catheters or on sections of coated tubing from catheters (eg, 6 cm long).

[0148] 1. Apply a generous amount of lidocaine gel (or similar) between the thumb and index finger of a dry, gloved hand.

[0149] 2. To hold the coated catheter (or coated catheter fragment), gently pinch the coated catheter between your index finger and thumb using a gloved hand coated with lidocaine gel (or similar). Gently rub the surface of the coated catheter longitudinally 10 times within a 4 cm area.

[0150] 3. After abrasion, the catheter or catheter segment should be rinsed with deionized water to remove excess gel from the surface.

[0151] 4. Subject the abraded coated catheter (or coated catheter fragment) to the dye test described herein to visually assess the coating's resistance to damage and minor abrasion compared to a coated control (no gel abrasion).

[0152] Bending test This was done on coated catheters or on sections of coated tubing from catheters (eg, 6 cm long).

[0153] 1. To hold the coated catheter (or coated catheter fragment), use dry, gloved hands to gently bend and twist the coated catheter 10 times.

[0154] 2. Subject the coated catheter (or coated catheter segment) to the dye test described herein to visually assess the coating's resistance to damage and bending and twisting compared to a coated control (no bending or twisting).

[0155] Pigment test The dye test described herein was performed by immersing catheters or catheter tubing segments in an aqueous solution containing crystal violet for 60 seconds. The catheters or catheter tubing segments were then removed from the aqueous solution, immersed in deionized water for 60 seconds, and then removed from the solution. The color consistency, or uniformity of the color, was then visually evaluated. The coated areas were stained purple / dark blue, while the uncoated areas did not absorb the dye and therefore remained uncolored. The quality of the coating was also visually evaluated to determine whether there was any cracking, flaking, and / or delamination of the coating.

[0156] Two replicates were performed for each test method.

[0157] The results were as shown in Table 2. [Table 2]

[0158] As shown in Table 2, the coatings did not abrade in these tests. It was concluded that the coated medical devices described herein, such as the catheters described herein, are abrasion resistant. Thus, the coatings on the medical devices described herein resist abrasion, such as abrasion during use.

[0159] Example 4: Cytotoxicity test The catheter segments prepared according to Example 1 were subjected to an MTT cytotoxicity test in accordance with ISO standard ISO10993-5:2009. MTT is the salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. In this test, the yellow, water-soluble MTT is metabolically reduced to a blue-purple, insoluble formazan in viable cells. The number of viable cells correlates with the intensity of the color, determined photometrically after dissolving the formazan in alcohol. No cytotoxicity was observed when the catheter segments were subjected to the MTT test.

[0160] Example 5: Determination of the amount of PDEA in the coating To estimate the amount of bound PDEA in the coated catheter prepared in Example 1, L-cysteine ​​was added to the coated sample in solution. When L-cysteine ​​reacts with PDEA, 2-mercaptopyridine is released, which can be measured by UV spectrophotometry (342 nm). See Scheme A below. [ka]

[0161] As long as the reaction is complete, i.e., all PDEA has reacted with L-cysteine, the amount of 2-mercaptopyridine released represents the amount of PDEA present in the coating. This reaction also occurs when PDEA is non-covalently (e.g., ionically) bound (scheme not shown).

[0162] To ensure complete reaction, fresh L-cysteine ​​solution was added every 10 minutes until no further 2-mercaptopyridine was detected and the reaction was deemed complete. 1. Coated samples were prepared from FR-16 size coated catheters. Two 5 cm catheter pieces were cut in half to give four samples with a total coated area of ​​13.5 cm2 / sample. 2. 100 ml of L-cysteine ​​solution (0.072 M) was prepared. 3. 5 ml of L-cysteine ​​solution was added to the sample to initiate the reaction. 4. The L-cysteine ​​solution was removed after 10 minutes and 2-mercaptopyridine was measured by UV at 342 nm. 5. Steps (3) and (4) were repeated until 2-mercaptopyridine was no longer detectable.

[0163] The cumulative amount of 2-mercaptopyridine measured, and therefore of PDEA reacted, is shown in FIG.

[0164] A total of 12 consecutive 10-min coupling reactions were performed. The total amount of PDEA present in the coating was 2.22 nmol / cm 2 Approximately half of the total PDEA (1 nmol / cm) was determined to be in the first 10 min. 2 ) reacted with L-cysteine, and after 10 couplings, no additional 2-mercaptopyridine was detected.

[0165] Reference Example 6: PDEA content in coating of previous generation products The previous coating specifications were applied to silicone nasal prongs of the type used in the first-in-man study (Odeberg et al., supra). The purpose of this experiment was to measure the amount of PDEA and cysteine ​​ligands present in the previous generation coating.

[0166] The main component of the previous coating was polyacrylic acid, and the previous coating used free radical electron beam (EB) induced polymerization instead of the UV induced polymerization used in the above examples.

[0167] The amount of PAA in the previous coating was approximately 4–15 μmol / cm 2 It was.

[0168] PDEA was covalently attached to the surface following a similar method to that described in Example 1, except that higher concentrations of EDC / NHS and PDEA were used.

[0169] After PDEA coupling, the cysteine ​​ligand was coupled according to Scheme B. [ka]

[0170] The amount of remaining unreacted PDEA was determined using the same method as in Example 5, except that 18 consecutive coupling reactions were performed.

[0171] The cumulative amount of 2-mercaptopyridine measured for the PDEA-coupled intermediate, and therefore the PDEA reacted, is shown in FIG.

[0172] The 10-minute data point represents the manufacturing procedure. Therefore, due to the high amount of PDEA and the relatively short L-cysteine ​​reaction time (10 minutes), not all of the PDEA was consumed in the previous coating process. Therefore, the previous coating contained both covalently bound PDEA and cysteine ​​ligands.

[0173] As can be seen from Figure 3, the total amount of PDEA ligand coupled in the production process was approximately 257 nmol / cm 2 It was.

[0174] The measured cumulative amount of 2-mercaptopyridine, representing unreacted PDEA, after the first 10 minutes of cysteine ​​coupling is shown in Figure 4. These results suggest that the remaining amount of PDEA ligand after the completion of the manufacturing process is at least about 65 nmol / cm. 2 It was estimated that:

[0175] In summary, the previous coatings were approximately 74% (191 nmol / cm 2 ) cysteine ​​ligand and 26% (66 nmol / cm 2 ) contained PDEA and

[0176] Compared to the coating of Example 1, the previous coating had a 2.2 nmol / cm 2At least 25 times more bound PDEA compared to PDEA, i.e., >65 nmol / cm 2 It contained:

[0177] Example 7: Dose response to various amounts of PDEA The purpose of this experiment was to determine a dose-response curve for different levels of bound PDEA in the coating. The limit of PDEA at which the coating will inhibit bacterial growth is of particular interest because the mode of action changes from anti-biofilm coating formulations that do not rely on bacterial growth inhibition (the present invention) to previous "bacterial growth inhibition" coating formulations.

[0178] Silicone-coated catheters, size Fr14, were grafted with PAA and the samples were cut into 2.5 cm pieces. By titration, the amount of PAA was determined to be approximately 6.5 μmol / cm 2 It was determined that (see Example 1).

[0179] Coupling was performed using the same EDC / NHS concentrations but varying the PDEA concentration as in Example 1. The amount of bound PDEA was determined by UV (342 nm) measuring the released 2-mercaptopyridine after reacting the sample with L-cysteine ​​(see Example 5 for method description). The results are shown in the table below. [Table 3]

[0180] Antibacterial Assay - Ahearn Test Bacterial strain: Escherichia coli CFT073 ·Starting inoculum: 15×10 5 CFU / ml Culture medium: Artificial Urine Medium (AUM) Three replicates were used for each test sample.

[0181] The Ahern test procedure was used to determine bacterial colony forming units on the surface and in the AUM after 3 hours of incubation with bacteria. 1. Test samples (2.5 cm, Fr 14, total surface area 5.8 cm2) were incubated in phosphate buffer overnight under ambient conditions with slow rotation. 2. The test piece was placed in a 2 ml tube. 3.E.coli (15×10 5 Two ml of AUM containing 100 CFU / ml was placed in the tube. 4. These were incubated at 37±2°C for 3 hours. 5. Samples were taken for dilution. 6. The samples were washed twice with 2 ml each of PBS to remove loosely attached bacteria and the bacterial suspension was discarded. 7. After washing, the sample was placed in a new 2 ml microcentrifuge tube. 8. To recover the attached bacteria, 1 ml of PBS containing 0.05% Tween-20 was added to each tube, and the tubes were vortexed for 30 seconds, then sonicated for 5 minutes (40 kHz in an ultrasonic bath Bransonic at 100 W power under ambient conditions) and vortexed again for 30 seconds. 9. Serial dilutions were prepared in PBS and spread onto LA plates with glass beads and plate counted. Dilutions 0, -10, -100 were plated.

[0182] The results are shown in Figure 5 (attached bacteria) and Figure 6 (bacteria in the medium). 2 A dose-response relationship is obtained showing that in the range above 6 nmol / cm, significant inhibition of bacteria begins both on the catheter surface and in artificial urine medium (AUM). In contrast, lower amounts of bound PDEA have no inhibitory effect on bacterial growth. In contrast, at 6 nmol / cm compared to the control. 2 Sample and 13 nmol / cm 2 In both samples, there are indeed slightly higher numbers of attached bacteria.

[0183] The half-maximal inhibitory concentration (IC50) of bound PDEA was estimated to be 39 nmol / cm on the surface. 2 , 30 nmol / cm in AUM 2 (interpolated value).

[0184] Bound PDEA is 77 nmol / cm 2 The log reduction is 1.55 log CFU / cm 2 and 2.77 log CFU / ml, which is consistent with the estimated amount of bound PDEA in a previous patent application (WO 2006 / 101438 A1) and in prior art coatings (65 nmol / cm 2 It matches super.

[0185] In conclusion, the "no bacterial growth inhibition" range for bound PDEA is 30 nmol / cm 2 It is defined as less than.

[0186] Example 8: The coating of the present invention has no metabolic or pharmacological effects. The purpose of this experiment was to demonstrate that the coating of the present invention (referred to herein as CytaCoat) has no metabolic or pharmacological effect on bacteria. The following gram-positive and gram-negative bacteria were included in the study: >Klebsiella pneumoniae AO15200 (clinical isolate) > Staphylococcus epidermidis Se19 (clinical isolate)

[0187] This study used uncoated and coated silicone size Fr20 catheters: >Untreated catheter, 2 cm piece ("control") >CytaCoat catheter sample, 2 cm piece ("Cytacoat").

[0188] The "CytaCoat" material was prepared as described in the previous example, so that the coating contained approximately 2.2 nmol / cm 2 had PDEA.

[0189] The experimental setup is illustrated in FIG.

[0190] Week 1: 1. Six catheter controls and six CytaCoat catheter samples were used in this study.

[0191] 2. On the first day of the test, 9 One ml of freshly prepared artificial urine (AUM) solution containing cfu / ml of K. pneumoniae or S. epidermidis cells was added to the tube.

[0192] 3. Samples were incubated at 37°C for 7 days, with AUM replaced daily except on weekends.

[0193] 4. When the AUM was changed, the sample was washed twice in 1.5 ml of PBS in a new microcentrifuge tube and transferred to 1.5 ml of fresh AUM in a new microcentrifuge tube.

[0194] On day 5 and day 8, three samples were collected for crystal violet biofilm quantification and fixed as follows: a) 1% glutaraldehyde in PBS for 5 min; b) 60% ethanol for 15 min; c) 80% ethanol for 15 min; and d) dried.

[0195] 6. These samples were stained with 0.04% crystal violet in water for 5 minutes, then rinsed in Milli-Q water and dried. The biofilm was removed with a clean cotton swab, and the crystal violet was extracted with 1.5 ml of 95% EtOH for 3 hours, and the OD595nm was measured (Figure 8).

[0196] 7. Aliquots were taken from the tubes containing the other three samples for serial dilutions and colony counts to assess colony forming units in liquid cultures (Figure 7).

[0197] 8. These samples were washed twice in 1.5 ml of PBS in a new microcentrifuge tube and transferred to 1.5 ml of fresh AUM in a new microcentrifuge tube. Biofilm was removed from these samples and disrupted by 30 seconds of treatment with a handheld ultrasonic homogenizer mixer. Aliquots were taken for serial dilution and colony counting to assess colony-forming units on the catheters (Figure 7).

[0198] 9. The resulting biofilm bacteria in the AUM were transferred to fresh Eppendorf tubes containing both control catheter pieces, and the experiment continued for another week (see Week 2 below).

[0199] Week 2: 10. Samples from the previous week's experiment were incubated at 37°C for 7 days with daily AUM changes except on weekends.

[0200] On day 11.15, two samples were taken for the crystal violet biofilm quantification procedure. They were fixed as follows: a) 1% glutaraldehyde in PBS for 5 min; b) 60% ethanol for 15 min; c) 80% ethanol for 15 min; and d) dried.

[0201] 12. These samples were stained with 0.04% crystal violet in water for 5 min, then rinsed in Milli-Q water and dried. The biofilm was removed with a clean cotton swab, and the crystal violet was extracted with 1.5 ml of 95% EtOH for 3 h, and the OD595nm was measured (Figure 8).

[0202] 13. Control and one CytaCoat sample were selected for microscopic visualization. Sample sections were stained with Ebba Biotech Red (680 nm) / Concanavalin A-Alexa probe by the following method. a) A working solution of dye was prepared as follows: 1 μl Ebba Biolight Red (630 nm) and 1 μl Concanavalin A, Alexa Fluor™ 594 conjugate (50 μg / ml). b) The specimen was placed in 1 ml of staining solution and incubated in the dark at room temperature for 30 minutes. c) The stained specimens were briefly rinsed in water and further washed with 1 ml of water in the dark. d) After staining, the specimens were visualized under a fluorescent microscope.

[0203] Consideration After week 1, both K. pneumoniae and S. epidermidis formed EPS / biofilm on the control, while CytaCoat catheter samples showed little detectable biofilm. After week 2, when pre-exposed bacteria were transferred from week 1 to the control catheter, they formed even more EPS / biofilm compared to week 1. Biofilm formation on the control surface at week 2 was independent of whether the bacteria were exposed to a CytaCoat or control catheter at week 1.

[0204] conclusion While preventing normal biofilm formation, the CytaCoat surface does not impede the growth of K. pneumoniae AO15200 or S. epidermidis Se19. Additionally, bacteria growing on the CytaCoat surface show no change in their properties and retain the ability to form biofilms when transferred to a control silicone catheter surface.

[0205] These results indicate that the CytaCoat coating creates a local anti-biofilm environment and that it has no metabolic or pharmacologically supportive effect on the bacteria.

[0206] References 1. International Publication No. 2006 / 101438 A1 2.Letters in Applied Microbiology 1997,24,203-206 3.Jacob Odeberg,Anders Wirsen,Ake Norberg,Jakob Frie,Gordana Printz,Hugo Lagercrantz,Gudmundur H Gudmundsson,Birgitta Agerberth,Baldvin Jonsson;A novel cysteine-linked antibacterial surface coating significantly inhibits bacterial colonization of nasal silicone prongs in a phase one pre-clinical trial.Mater Sci Eng C Mater Biol Appl 2018 Dec 1;93:782-789

Claims

1. 1. A medical device comprising a coating, the coating comprising: Polymer Z attached to the medical device and the moiety of formula A 【Chemistry 1】 wherein X is an amino group covalently bonded to Z or capable of ionically bonding to a free carboxyl group. Including, The moiety according to Formula A is present in the coating at a concentration of 0.5 to 30 nmol / cm ２ is present in an amount of Polymer Z is 1 to 30 μmol / cm ２ containing free carboxyl groups in an amount of Medical equipment.

2. The coating has a structure of Formula I: 【Chemistry 2】 (I) wherein the circle represents the surface of the medical device.

10. The medical device of claim 1, comprising:

3. The coating is a compound in which X is a primary amino group -NH ２ , a secondary amino group -NRH, or a tertiary amino group -NR ２ and each R is C １～４ 3. The medical device of any one of claims 1 to 2, comprising a moiety of formula A independently selected from alkyl.

4. 10. The coating according to claim 1, wherein the coating has a structure: 【Transformation 3】 4. The medical device of claim 1 , comprising:

5. 5. The medical device of claim 1, wherein at least 20 mol. % of the moieties according to formula A are attached to Z via covalent bonds.

6. 6. The medical device of claim 1, wherein at least 20 mol. % of said moieties according to Formula A are ionically associated with said coating.

7. The moiety according to Formula A is present in the coating at 0.5 to 15 nmol / cm ２ 7. The medical device of claim 1, wherein the amount of

8. Polymer Z is 3 to 7 μmol / cm ２ 8. The medical device of claim 1, comprising free carboxyl groups in an amount of

9. The coating 2-(pyridyldithio)ethylamine (PDEA) covalently attached to a polymer that is covalently attached to the medical device. Including, 9. The medical device of claim 1, wherein the PDEA is present in an amount of 0.001 wt% to 1 wt% based on the total weight of the coating.

10. 10. The medical device of claim 9, wherein the PDEA is present in an amount of 0.01 wt% to 1 wt% based on the total weight of the coating.

11. 11. The medical device of claim 1, wherein the polymer Z is covalently attached to the medical device.

12. 12. The medical device of any one of claims 1 to 11, wherein the coating comprises a polymer selected from the group consisting of polyacrylic acid, polymethacrylic acid, poly-4-vinylbenzoic acid, polyitaconic acid, any combination thereof, and esters or amides thereof.

13. 13. The medical device of any one of claims 1 to 12, wherein the polymer Z comprises an acrylate polymer or an acrylic polymer, such as polyacrylic acid.

14. 14. The medical device of claim 1, wherein the polymer Z of the coating is grafted from the medical device.

15. 15. The medical device of any one of claims 1 to 14, wherein the coating comprises water, thereby forming a hydrogel.

16. 16. The medical device of any one of claims 1 to 15, selected from the group consisting of a catheter, an implant, a tracheal tube, a stent, a ventilator, a wound dressing, a face mask, a nasal prong, a hearing aid, and a syringe.

17. 17. The medical device of any one of claims 1 to 16, selected from the group consisting of a central venous catheter, a ureteral stent, a wound dressing, a ventilator, a face mask, a nasal prong and an implant.

18. 18. The medical device according to any one of claims 1 to 17, which is a catheter, such as a Foley catheter, or an endotracheal tube.

19. polymers, such as thermoplastic and / or thermosetting polymers, and / or metal 19. The medical device of any one of claims 1 to 18, comprising:

20. the polymer comprises one or more of silicone, polyethylene, polypropylene, polyurethane, polyvinyl chloride, polycaprolactone, polycarbonate, rubber such as latex rubber, polyetheretherketone (PEEK); and / or the metal comprises one or more of a steel such as stainless steel, an alloy such as a cobalt-based alloy or nitinol, titanium; 20. The medical device of claim 19.

21. 21. The medical device of any one of claims 1 to 20, wherein the coating is present on at least a portion of the interior and / or exterior surface of the medical device.

22. 22. A medical device according to any one of claims 1 to 21, which is free or substantially free of biofilm after 4 days or more, such as 30 days, of clinical use.

23. 23. A coating for a medical device as defined in any one of claims 1 to 22.

24. 1. Use of a coating to reduce the formation and / or growth of biofilms on a surface of a medical device, said coating comprising: Polymer Z attached to the medical device and the moiety of formula A 【Chemistry 4】 wherein X is an amino group covalently bonded to Z or capable of ionically bonding to a free carboxyl group. Including, Polymer Z is capable of forming a hydrogel when in contact with a fluid containing water; use.

25. 25. The use of claim 24, wherein the polymer Z is covalently attached to the medical device.

26. 26. The use according to claim 24 or claim 25, wherein Z comprises a free carboxyl group.

27. Z is 1 to 30 μmol / cm ２ 27. The use according to claim 26, wherein the amount of free carboxyl groups is

28. Z is 3 to 20 μmol / cm ２ 28. The use according to claim 27, wherein the amount of free carboxyl groups is

29. The coating has a structure of Formula I: 【Transformation 5】 wherein the circle represents the surface of the medical device. The use according to any one of claims 24 to 28, comprising:

30. The coating is a compound in which X is a primary amino group -NH ２ , a secondary amino group -NRH, or a tertiary amino group -NR ２ and each R is C １～４ 30. The use of any one of claims 24 to 29, comprising a moiety of formula A independently selected from alkyl.

31. 10. The coating according to claim 1, wherein the coating has a structure: 【Transformation 6】 The use according to any one of claims 24 to 30, comprising:

32. 32. The use according to any one of claims 24 to 31, wherein at least 20 mol. % of said moieties according to formula A are attached to Z via a covalent bond.

33. 33. The use according to any one of claims 24 to 32, wherein at least 20 mol. % of said moieties according to formula A are ionically associated with said coating.

34. The moiety according to Formula A is 0.5 to 30 nmol / cm ２ The use according to any one of claims 24 to 33, wherein the composition is present in the coating in an amount of

35. The moiety according to Formula A is 0.5 to 15 nmol / cm ２ 35. The use of claim 34, wherein the coating is present in an amount of

36. 36. The use of any one of claims 24 to 35, wherein the coating comprises a polymer selected from the group consisting of polyacrylic acid, polymethacrylic acid, poly-4-vinylbenzoic acid, polyitaconic acid, any combination thereof or esters or amides thereof.

37. The use according to any one of claims 24 to 36, wherein the polymer Z comprises an acrylate polymer or an acrylic polymer, such as polyacrylic acid.

38. The use according to any one of claims 24 to 37, wherein the polymer Z of the coating is grafted from the medical device.

39. The use according to any one of claims 24 to 38, wherein the coating is as defined in any one of claims 1 to 22.

40. Use according to any one of claims 24 to 39, wherein the reduction is not accompanied by a bactericidal or bacteriostatic effect.

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