Photosensitizer, method for obtaining functional polymer surface using functional polymer material, and use thereof

Abstract

The present invention relates to a method for obtaining functional polymer surfaces (MPn-PSm) from polymers or copolymers with appropriate functional groups (X). Examples of functional groups (X) include polyvinyl chloride (MP1), (chloromethyl)polystyrene-Merrifield (MP2), curcumin photosensitizers (PS1), meso-tetra(aryl)porphyrins, chlorins, or basophilic groups (PS2), particularly halogen-containing leaving groups such as P2 in meso-imidazolyl-porphyrins, chlorins, or bacteriochlorins (PS3). All photosensitizer structures in this application incorporate a functional group (Y) that is a nucleophile, such as OH, SH, or NH2. The present invention also relates to a method for covalently bonding photosensitizers PS1-PS3 to functional polymer materials MP1-MP2 through a nucleophilic substitution reaction to produce MPn-PSm products. The resulting products can prevent the growth of microorganisms that can cause infections, a major cause of death in device-using patients.

Description

[Technical Field]

[0001] The present invention relates to the field of polymer functionality. It primarily consists of a method for attaching photosensitizing molecules to polymer surfaces, each containing appropriate chemical functional groups that allow for thermodynamically stable covalent chemical bonds, resulting in a pair of photosensitized polymer materials (MPn-PSm). These functional materials, when irradiated in the dark or, preferably, at an appropriate wavelength, promote photodynamic action and, when present in a medium, can prevent the formation of surface microbial colonies while eliminating microorganisms that come into contact with the surface.

[0002] More specifically, it applies to polymeric biomedical devices used in life support, such as valves, packaging, endotracheal tubes with or without device storage bags, catheters, probes, reservoirs, gloves, tracheostomy tubes, intravenous scalpels, nasal oxygen catheters, and tracheal suction tubes. [Background technology]

[0003] Immunosuppressed patients using these devices in hospitals typically receive standard antibiotic treatments for infections, leading to the development of bacteria that are multidrug-resistant to most antibiotics on the market.

[0004] Endotracheal tubes are a common device used to support mechanical ventilation in patients with respiratory disease, post-surgery, or trauma. Microbial colonies commonly grow on the surface of the tube, leading to the development of ventilator-associated pneumonia. According to data from Zeitoun, SS et al. (2001), the risk of death from microbial infection is 21 times higher in intubated patients than in non-intubated patients, making endotracheal tubes one of the leading causes of death in intubated patients.

[0005] Blood bags are devices used to transport and store donor blood until transfusion. Blood transfusion contamination is a very serious problem and can cause systemic infection in patients.

[0006] The transport of transplant organs must be carried out in packaging that does not pose a risk of microbial contamination and does not alter the integrity of the organ, so ensuring the sterility of medical equipment for transport is of paramount importance.

[0007] This invention describes a process for conjugating photosensitizers (PS) containing components of curcumin groups and derivatives (PS1), porphyrins and derivatives (chlorins, bacteriochlorins) (PS2 and PS3) to generate stable covalent chemical entities (ethers, amines, thioethers) with polymer surfaces (MP1 and MP2) containing appropriate functional groups (X) to generate (MPn-PSm) products.

[0008] The product developed in this invention (MPn-PSm) is stable in aqueous solutions of different pH after irradiation with light of appropriate wavelengths at physiological temperature.

[0009] Products with covalently bonded photosensitizers (PS) (MPn-PSm) functionalized polymer surfaces can prevent the onset of microbial infections and reduce the risk of death in the absence of light. Furthermore, when exposed to light of the appropriate wavelength, these products can inactivate microorganisms or biofilm formation through photodynamic action, thereby reducing the number of deaths caused by multidrug-resistant microbial infections.

[0010] Inactivation of microorganisms by photodynamic therapy (PDT) requires three components: a photosensitizer (PS), a light source of the appropriate wavelength, and oxygen.

[0011] Photosensitizers (PS) are chemicals that absorb light of specific wavelengths and change its chemical and / or physical properties. PS absorb energy from light by entering an excited state and transferring the energy. They can operate via two distinct mechanisms: Type I and Type II. In the Type I mechanism, PS react directly with molecules, generating reactive free radicals (ROS) and radical ions. In the Type II mechanism, energy is transferred to molecular oxygen, generating singlet oxygen, which can inactivate microorganisms (Pucelik et al. (2018) (PlosOne, 13(1):e0191777).

[0012] However, the development of microbial resistance due to the action of reactive oxygen species (ROS) generated by photodynamic therapy (PDT) has not yet been reported.

[0013] Bezman et al. (1978) (Photochemistry and Photobiology, 28, 325-329, 1978) reported the photodynamic inactivation of Escherichia coli using the photosensitizer (PS) Rose Bengal covalently bonded via an ester bond to beads of a copolymer of polystyrene and chloromethylstyrene. Furthermore, this ester covalent bond is susceptible to hydrolysis in biological media and the action of light. On the other hand, the preparation method or MPn-PSm product described in this invention allows the formation of stable covalent bonds (amine, ether, thioether) between different photosensitizers (PS) with selected functional groups (NH, OH, SH) and different polymers, such as PVC and Merrifield, which are stable to light irradiation in biological media.

[0014] Patent WO1993 / 000815 discloses a photobactericidal composition for surface sterilization, which contains a fiber-type polymer (cellulose) and a photosensitizer (porphyrin or phthalocyanine) linked solely by electrostatic interactions. The photosensitizer used is a non-functional mesotetrapyridylporphyrin or phthalocyanine type, which is adsorbed to the polymer solely by electrostatic bonding.

[0015] EP1203052 / US6420455B1 discloses a polymer composition and a product using it that exhibits surface antibacterial properties. However, the presence of a covalent bond with a photosensitizer or even xanthene was not mentioned. The interaction between the photosensitizer and the polymer material was merely a purely physical interaction (physical mixing) involving some electrostatic interactions, rather than a fundamental chemical interaction. The document also describes the drug's effect during periods without light stimulation, showing a significant difference in the interaction due to the mechanical requirements and other functions of the base polymer material being maintained, but not the biological surface properties maintained by the surface functionalization of the present invention.

[0016] Document RU2663061 relates to polymer-based antimicrobial agents with bactericidal properties. Their molecules contain at least one nitrogen atom with a pair of free electrons, allowing the biocide to coordinate to metal complexes through unstable electrostatic bonds. These complexes may contain magnesium porphyrin as a central atom, in addition to other chemical groups such as phthalocyanines. The only similarity with the present invention is the use of porphyrin-type photosensitizers, but with different structures and resulting photodynamic properties. Furthermore, the polymers described in this document are not intended for the functionalization of biomedical device surfaces through covalent bonding.

[0017] The novelty of this invention is based on the functionalization of the polymer surface of polyvinyl chloride, halomethylpolystyrene, or their copolymers with leaving groups (Br, Cl, I, or F), particularly chlorine (Cl) leaving groups, PVC (MP1) or Merrifield (MP2), which bind to photosensitizers. These compounds have appropriate functional groups (OH, N-, or SH) that, in the presence of light in biological media, form stable covalent bonds to form products such as MPn-PSm upon irradiation with light of the appropriate wavelength, which can be used in biomedical devices with antibacterial properties.

[0018] It is particularly important to note that the materials obtained by the present invention significantly reduce the risk of infection and pneumonia in hospitalized and intubated patients. Therefore, functionalized polymer materials with covalently attached photosensitizers (PS) can bring significant benefits to public health, such as improving the hospital environment. Summary of the Invention [Problem to be solved by the invention]

[0019] The present invention relates to a method for preparing polymeric products (MPn-PSm) containing appropriate functionalized polymers or copolymers. The polymeric products (MPn-PSm) developed in this invention reduce microbial growth in the absence of light and exhibit antimicrobial activity when exposed to light of an appropriate wavelength. Polymeric materials (MPn-PSm) bound to photosensitizers (PSm) via irreversible covalent bonds have demonstrated microbial inactivation functionality in medical devices, endotracheal tubes, organ storage and transport bags, blood storage and transport bags, and food storage and transport bags. [Brief explanation of the drawings]

[0020] For a complete understanding of the subject matter of this patented invention, please refer to the following diagram: [Figure 1] Figure 1 shows a functionalization scheme for a polymeric material (MPn) containing a leaving group (X), which may be a halogen atom, reacting with a photosensitizer (PSm), which may be a curcumin-type derivative, a tetrapyrrole macrocycle (porphyrin, or derivative (chlorin or bacteriochlorin) with a nucleophilic group (Y). The nucleophilic group (Y) may be of the type -OH, -N-, or -SH, generated from the functionalized polymeric material (MPn-PSm). [Figure 2] Figure 2 shows control data for the polymeric material of the tube (MP1) (black curve) and the curcumin photosensitizer (PS1) (blue curve) in a trachea functionalized with curcumin photosensitizer (MP1-PS1) (green). [Figure 3]Figure 3 shows examples of infrared spectroscopy (FT-IR) characterization of a functionalized endotracheal tube (PVC-curcumin, MP1) (red curve) and a functionalized endotracheal tube (ET) (black curve). Infrared spectroscopy characterization was measured in the range of 500-4000 cm-1 on a spectrophotometer with a Smart Orbit accessory. [Figure 4] Figure 4 shows examples of scanning electron microscopy (SEM) characterization. (a) An endotracheal tube composed of a solid-state polymer material (MP1). (b) An endotracheal tube composed of a curcumin (PS1) (MP1-PS1) covalently functionalized polymer material (MP1). SEM images were obtained with a ZEISS LLE O440 (Cambridge, England) instrument equipped with an OXF or D detector (model no. 7060) using a 15 kV electron beam, a 2.82 A current, and a 200 pA AI probe. The samples were coated with 6 nm of gold using a BAL-TEC MED020 Coating System Metallizer (BAL-TEC, Liechtenstein) and stored in a desiccator until analysis. Metallization conditions were: chamber pressure = 2.00 x 10-2 mbar; current = 60 mA; deposition rate = 0.60 nm / s. [Figure 5] Figure 5 shows an example of fluorescence spectroscopy characterization of an endotracheal tube composed of a covalently bonded functional material (MP1-PS1) in the 350-600 nm excitation wavelength range. a) Fluorescence image acquired from the surface of the functionalized endotracheal tube (MP1-PS1). [Figure 6] Figure 6 shows the microbiological analysis results for the reduction of S. aureus biofilm on curcumin-functionalized endotracheal tubes by covalently bonded MP1-PS1. Orange ( / / / ) indicates the percentage reduction in bacterial growth of biofilm on TE endotracheal tubes (control) and curcumin-functionalized endotracheal tubes by covalently bonded MP1-PS1 (orange) in the dark. Red ( / / / ) indicates the percentage reduction in bacterial growth of biofilm on TE endotracheal tubes (control) and curcumin-functionalized endotracheal tubes by covalently bonded MP1-PS1 (red) after 12 minutes of irradiation with a total dose of 50 J / cm2 at 450 nm using an LED light source. [Figure 7] Figure 7 shows a graph of the UV-Vis spectra of the polymer material MP1-PS1 immersed in solutions under various pH conditions (2, 7, 10).

[0021] Detailed Description of the Invention The present invention relates to functionalizing polymeric and copolymeric materials by covalently bonding photosensitizers (PSs) in the dark, preferably under light irradiation (PDT), to reduce microbial adhesion and / or inactivate biomedical devices fabricated from these materials. These polymers are preferably from the vinyl polyhalide class. Halides or halogens are diatomic molecules of elements from Group 17 of the periodic table, containing the leaving group fluorine (F), chlorine (Cl), bromine (Br), or iodine (I), preferably chlorine (Cl).

[0022] The present invention relates to a method for obtaining functional polymer surfaces (MPn-PSm) from polymers or copolymers bearing appropriate functionalities (X). This functionality (X) consists of halogen or leaving groups, in particular polyvinyl chloride (MP1) and (chloromethyl)polystyrene-Merrifield (MP2), curcumin-type photosensitizers (PSs) and derivatives (PS1), bacteriochlorins, meso-tetraaryls, porphyrins and derivatives, in particular meso-imidazoyl-porphyrin type and derivatives (chlorins and bacteriochlorins) (PS3), halogenated and functionalized with chlorine or nucleophilic groups (PS2). All photosensitizers (PSs) of the present invention incorporate a functional group (Y) in their structure, which is a nucleophile of the OH, SH or NH type.

[0023] Specifically, the present invention relates to a method for producing MPn-PSm substances by covalently bonding (D) ether, amine, or thioether types of PS1, PS2, and PS3 to functional polymer materials MP1 and MP2 through a nucleophilic substitution reaction. Furthermore, the resulting MPn-PSm substances enable the inactivation and reduction of microbial adhesion rates through photodynamic therapy (PDT). These substances are applicable to human and veterinary hospital facilities and devices, such as probes, catheters, reservoirs, tracheostomy tubes, intravenous infusion devices, oxygen catheters, hemodialysis catheters, rectal probes, organ transport and storage packages, urethral probes, and tracheal suction probes.

[0024] The product formed on the surface of the MPn-PSm polymer developed in this invention can prevent microbial growth. Microbial growth is one of the leading causes of death in hospitalized patients and is also the cause of many serious infections. These MPn-PSm polymer materials are also finding great applications in the widespread use of food storage and packaging.

[0025] The method for obtaining these is simple: MPn-PSm (n1-2, m1-3, i.e., MP1-PS1, MP1-PS2, MP1-PS1; MP1-PS2; MP1-PS3; MP2-PS1; MP2-PS2, and MP2-PS3, see Table 1). The MPn-PSm is obtained by direct nucleophilic substitution of the leaving groups of polymer materials with halogens (fluorine, chlorine, bromine, iodine) and the respective PS1-type photosensitizers, PS2, and lipophilic groups, OH, SHN-, PS3.

[0026] The structures of the polymer materials MP1 (polyvinyl chloride (PVC) with the general formula [C2H3Cl]n) and MP2 ((chloromethyl)polystyrene with the general formula [C9H9Cl]n) allow easy identification of the presence of halogen leaving groups.

[0027] [ka]

[0028] Origin of photosensitizers The photosensitizer PS1, which has a hydroxyl-type nucleophilic group with nucleophilic function, is abbreviated as curcumin. Its IUPAC name is (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione. Its derivatives:

[0029] [ka]

[0030] Photosensitizers of the PS2 and PS3 families include porphyrins (formulas II-IX, with two beta-pyrrole positions (---) in double bond form) functionalized with hydroxyl, amine, and thiol-type nucleophiles, and their reduced derivatives, chlorins (one position (---) in single bond form and the other position (---) in double bond form), and bacteriochlorins (both positions (---) in single bond form).

[0031] Symmetrical porphyrins (with two double bond positions (---) in formulas II-IV) were synthesized from four equivalents of a halogenated aldehyde according to the nitrobenzene or nitrobenzene-NaY method. In this method, four equivalents of a halogenated aldehyde containing a nucleophile Y (OH, N-, or SH) are mixed. The nucleophile Y (OH, N-, or SH) is at the other position, and a mixture of acetic acid, propionic acid, and nitrobenzene is used as the solvent at 100-140 °C in NaY zeolite, with or without a reusable catalyst.

[0032] After hot NaY filtration and cooling, the porphyrins are either precipitated directly from the reaction medium or purified by methanol or flash chromatography to obtain the symmetrical photosensitizers of the PS2 family (Formulas II-IV). Unsymmetrical halogenated porphyrins (Formulas V-VII) in the double bond (---) form are synthesized according to the nitrobenzene or nitrobenzene-NaY method, which involves mixing two equivalents of a halogenated aldehyde Y (OH, N-, or SH) with a nucleophile Y. In this method, the nucleophile (OH, N-, or SH) is in the other position, and the selected structure is selected. Two equivalents of formaldehyde or its respective acetal and four equivalents of pyrrole are used, and the mixture of acetic or propionic acid and nitrobenzene is used as the solvent at 100-140 °C under aerobic conditions, with or without a reusable catalyst, in NaY zeolite. After hot NaY filtration, the porphyrins are purified by preparative flash chromatography to obtain the asymmetrical photosensitizers of the PS2 family (Formulas V-VII).

[0033] [ka]

[0034] Under aerobic conditions, using a mixture of acetic acid or propionic acid with nitrobenzene as the solvent, with or without a reusable zeolite-type NaY catalyst, at 100-140 °C, symmetrical porphyrins (with two positions (---) in the form of a double bond) were synthesized according to the nitrobenzene or nitrobenzene-NaY method, which consisted of mixing four equivalents of 1-methyl-2-(2-methyl- ...

[0035] Unsymmetrical halogenated porphyrins (Formulas V-VII) in the double bond (---) form were synthesized according to the nitrobenzene or nitrobenzene-NaY method, which involves mixing two equivalents of formaldehyde or its acetal with a nucleophile Y, four equivalents of pyrrole, and two equivalents of formaldehyde or its acetal. Under aerobic conditions, a mixture of acetic or propionic acid and nitrobenzene is used as the solvent at 100-140 °C, with or without a reusable catalyst of the NaY zeolite type. After hot NaY filtration, the porphyrins are purified by preparative flash chromatography to obtain the asymmetric photosensitizers of the PS2 group (Formulas V-VII).

[0036] [ka]

[0037] Porphyrin-PS metal complexes of PS2 and PS3 (formulas II-VII, VIII, and IX) are prepared by mixing solutions of the respective porphyrins dissolved in a suitable solvent, preferably chloroform or DMF, with the appropriate metal salt (Zn(OAc)2, Pd(OAc)2, or AlCl3, 40-150°C). After the complexation reaction is complete, the reaction mixture is purified by successive washes with saturated aqueous sodium bicarbonate. If necessary, the metal complexes can also be purified by flash chromatography.

[0038] According to the method described by Pereira MM, one of the porphyrin-type precursors (Formulas II-IV) obtained by the previous method was mixed in the solid state with a small excess of p-toluenesulfonylhydrazine (15 equivalents) in a Schlenk tube. The mixture was then placed under vacuum at 0.1 bar for approximately 1 hour. The mixture was then heated to 120-140 °C for a time optimized for each substrate. The reaction mixture was dissolved in a minimum amount of organic solvent and washed successively with sodium hydroxide and water. The resulting solid was then dissolved in DME and FeCl3.6H2O (1 equivalent). Hydrogen peroxide (3% in water) was then slowly added. The reaction was stopped upon the disappearance of the bacteriochlorin absorption peak (approximately 750 nm). The corresponding chlorine atoms (single bond form at one position (---) and double bond form at the other position (---), Formulas II-VII) were purified by washing and flash chromatography.

[0039] Bacteriochlorins with two (---) positions as single bond forms of PS2 and PS3 were synthesized according to the method described by Pereira M.M. The porphyrin was mixed with excess p-toluenesulfonylhydrazine (40 equivalents) in a Schlenk tube and placed under vacuum (0.1 bar) for 1 hour. The mixture was then heated to 140 °C for a time optimized for the porphyrin (Formulas II–VII, where (---) represents a double bond). After cooling to room temperature, the corresponding bacteriochlorins (Formulas II–IX, where (---) represents a single bond) were purified by washing or flash chromatography.

[0040] Table 1 shows all possible bond structures between the polymeric material and the photosensitizer (MPn-PSm). The D component is always an ether (O), thioether (S), or amine (N-) covalent bond (Table 1):

[0041] Possible bonding and possible structures of polymer materials and photosensitizers (MPn-PSm)

[0042] [Table 1]

[0043] TIFF0007761230000006.tif222153

[0044] TIFF0007761230000007.tif191153

[0045] Table 2 shows possible substituents (R1, R2, Z, Z′, M) for the MPn-PSm bond described in Table 1.

[0046] MPn-PSm with substitutable R1, R2, Z, Z′, and M.

[0047] [Table 2]

[0048] TIFF0007761230000009.tif108137

[0049] Preparation of functional polymer products using MPn-PSm photosensitizer (PS) All PSs used in this invention (PS1, PS2, PS3) each have a specific chemical nucleophilic (OH, N-, or SH) structure. They can be easily and effectively linked to polymeric materials (MP1, MP2) through nucleophilic substitution reactions to form covalent bonds, incorporating halogen-type leaving groups into their structures. The specific method is as follows:

[0050] A photosensitizer (PS) selected from the PS1, PS2, or PS3 group is dissolved in an organic base (triethylamine, 1,8-diazabicyclo[5.4.0]und-7-ene (DBU), 1,5-diazabicyclo[4.3.0]non-5-ene (DBN), or pyridine) or preferably an inorganic base (CaCO3, CeCO3, NaOH, KOH, Ba(OH)3, Al(OH), Mg(OH), Be(OH), dimethylformamide (DMF), dimethoxyethane (DME), tetrahydrofuran (THF) and derivatives, dimethylpyrrolidone, dichloromethane, ethyl acetate, or preferably dimethyl sulfoxide (DMSO)) to form a SolA solution.

[0051] The polymeric materials MP1 and MP2 are immersed in the SolA solution and left at temperatures between 0°C and 40°C for 0.5 to 48 hours, preferably in the absence of oxygen. The covalently bonded polymeric materials are then removed to the desired photosensitizer (PS) (MPn-PSm) and washed 1-10 times with an organic solvent, preferably DMSO. Preferably, a low-boiling organic solvent such as ethanol is used.

[0052] MPn-PSm should be dried under vacuum at room temperature if possible. It is best to store it in an oxygen-free, light-shielded location.

[0053] The functional polymer products prepared in this invention (hereinafter referred to as MP1-PS1, MP1-PS2 (Formulas II-VII), MP1-PS3 (VIII-IX), MP2-PS1, MP2-PS2 (Formulas II-VII), and MP2-PS3 (VIII-IX)) were characterized by Cary 5000 UV-Vis diffuse transmittance spectroscopy to confirm the binding of the photosensitizer (PS) to the polymer material. The polymer material has a typical absorption band corresponding to each photosensitizer (PS). Details are shown in Figure 2.

[0054] As shown in Figure 3, the functional polymer materials MP1-PS1, MP1-PS2, MP1-PS3, MP2-PS1, MP2-PS2, and MP2-PS3 prepared by the present invention were characterized by infrared spectroscopy using a Nicolet 5700 (Thermo Electron Corporation) FTIR spectrometer (equipped with a Smart Orbit accessory), and the vibrational bands of specific functional groups associated with the bond between the polymer and each photosensitizer (PS) were observed.

[0055] The functional polymer materials MP1-PS1, MP1-PS2, MP1-PS3, MP2-PS1, MP2-PS2, and MP2-PS3 prepared according to the present invention were characterized by scanning electron microscopy (SEM) operating at a 15 kV electron beam, a 2.82 A current, and a 200 pA I probe. The samples were coated with 6 nm of gold and stored in a desiccator until analysis. Analysis revealed the presence of a photosensitizer (PS) on the polymer surface (Figure 4).

[0056] MP1-PS1, MP1-PS2, MP1-PS3, MP2-PS1, MP2-PS2, and MP2-PS3 were characterized by fluorescence spectroscopy on the outside of the tubes. Figure 5 shows representative curcumin fluorescence emission on the surface of material MP1-PS1. Inactivation of biofilm growth by MPn-PSm polymeric materials The MP1-PS1, MP1-PS2, MP1-PS3, MP2-PS1, MP2-PS2, and MP2-PS3 photosensitizer-functionalized polymer products prepared by the present invention inactivated bacterial biofilm growth. Here, bacterial biofilm refers to Gram-positive bacteria, particularly Staphylococcus aureus (Figure 6), or Gram-negative bacteria, particularly in the dark, preferably in an environment emitting light of the appropriate wavelength, i.e., in the visible range (370-700 nm) or near-infrared range (700-850 nm). Biofilm-forming microorganisms were incubated in suspension with polymer materials (MP1 or MP2) covalently bonded with selected photosensitizers (PS) PS1, PS2, or PS3 (MPn-PSm materials) for 24 hours. They were then exposed to an environment emitting light of the appropriate wavelength. To confirm the efficiency of photodynamic inactivation, a cell recovery method based on colony-forming units (CFU / mL) was used. The results demonstrated that the MPn-PSm polymer material was effective in reducing adhesion rates and inactivating microbial biofilm growth, regardless of whether it was irradiated with light of a wavelength appropriate for each photosensitizer (PS). Figure 6 in Example 1 shows the inactivation of microorganisms in the dark (56%) and after irradiation with light of a 450 nm wavelength (98%). This example demonstrates that the MPn-PSm material efficiently promotes the inactivation of dark biofilms through photodynamic action. Example 1: Method for preparing polymer material MP1-PS1 by covalently attaching curcumin (PS1) to an endotracheal tube (ET) made of PVC (MP1) Preparation of a solution of the present invention, SolA: Dissolve curcumin (396 mg; 1.07 mmol) and CsCO (1.99 g; 6.01 mmol) in dimethyl sulfoxide (DMSO; 80 mL).

[0057] An endotracheal tube (ET) (11 g) made of polymer material MP1 was weighed and immersed in Sol A at 30°C to 40°C for 4 to 8 hours in a nitrogen or argon atmosphere.

[0058] Remove the MP1-PS1 functional TE from the solution and wash it first with DMSO (4 times, 20 mL) and finally with ethanol (4-10 times, 20 mL) until no curcumin is observed by UV-Vis.

[0059] If possible, dry the functional TE (MP1-PS1) at room temperature in a vacuum environment for 1-3 days.

[0060] Store at room temperature, away from oxygen and light.

[0061] Characterization of the curcumin-functionalized TE tubes (PS1), designated as MP1-PS1 material, was carried out by various techniques.

[0062] UV-Vis: Figure 2 shows the UV-Vis absorption spectrum of the curcumin-functionalized endotracheal tube (Figure 2, green curve). At 430 nm, the spectrum shows the typical absorption spectrum of unimmobilized curcumin in ethanol solution (Figure 2, blue curve) and of the unabsorbed endotracheal tube (TE) (Figure 2, black curve).

[0063] FTIR: In Figure 3, the binding of curcumin in the endotracheal tube (TE) was verified by infrared contrast analysis of curcumin-functionalized tube (MP1-PS1, blue curve), endotracheal tube (THE, black curve) and curcumin (PS1, red curve). The spectrum of curcumin is shown at 3509 cm -1 (hydroxyl group -OH), 1600-1650cm -1 (C=O), 1509 cm -1 (C=C ethylene), 1250 cm -1The spectrum of the curcumin-functionalized endotracheal tube (MP1-PS1, blue curve) shows a peak at 3506 cm compared to the spectra of non-immobilized curcumin (PS1) and the endotracheal tube (TE). -1 (hydroxyl group -OH), 1600-1650cm -1 (C=O), 1512 cm -1 (COC ethylene), confirming the presence of covalently bound curcumin in the endotracheal tube (TE).

[0064] SEM: Scanning electron microscopy (SEM) examination of the endotracheal tube (ET) (Figure 4, a) and the curcumin-functionalized endotracheal tube (MP1-PS1) (Figure 4, b) confirmed the binding of curcumin to the endotracheal tube (ET) and demonstrated the presence of curcumin on the surface of the endotracheal tube (Figure 4, b). The MP1-PS1 material was designated to maintain the stability of the curcumin-functionalized TE tube (PS1).

[0065] The stability of the curcumin-functionalized endotracheal tube (MP1-PS1) was confirmed by analyzing its UV-vis absorption spectra over time at various pH values ​​(2, 7, and 10), simulating a biological system (Figure 7a-c). No release of curcumin from the endotracheal tube was observed. Curcumin Tube Microbiology The microorganism used was Staphylococcus aureus (ATCC 25925). The inoculum was prepared in a 15 mL Falcon tube containing brain heart infusion (BHI) growth medium and bacterial inoculum in a 9:1 ratio. The incubation time for the pre-inoculum was 15 hours at 37°C and 140 rpm in a rotary oven. Biofilm formation was performed as follows: - The pre-inoculated microorganisms were separated from the medium by centrifugation (1500 rpm for 15 min). -Two successive washes with phosphate-buffered saline (PBS) were performed by centrifugation (15 min at 1500 rpm). -Sterile endotracheal tubes were cut into 1 cm long pieces in a sterile environment (laminar airflow, Esco class IIbsc). Each section of the cut endotracheal tube was inserted into a separate well of a 24-well plate. - 900 μL of liquid medium and 100 μL of bacterial inoculum were added to the side of the well containing the cut tube. -The solution was homogenized by pipetting 1000 μL into each well of the plate 6 times. -The formed biofilms were characterized by bacterial colony counts (CFU / mL).

[0066] The illumination for the experiments was performed with an LED light source developed by the Laboratory for Technical Support - LAT / USP (San Carlos Physics Institute, IFSC / USP). The light source used in the MP1-PS1 irradiation experiments emitted radiation at 450 nm and had an output of 70 mW / cm. 2 The device is designed to uniformly and continuously irradiate a 24-well plate for 12 minutes. The irradiation measurement is performed on a 0.4 cm radius, 0.5 cm total area. 2 This was done using a potentiometer with a collector of 1 / 2 cm. The following equation was used to calculate the irradiance cm 2 was calculated. I=P / A; In the formula, I = potentiometer irradiance, P = power measured by the potentiometer, and A = area of ​​the potentiometer.

[0067] The illumination time required to achieve the required energy dose was calculated using the following formula: T=D / I; In the formula, T = illumination time, D = required energy dose, and I is the calculated irradiance of the LED.

[0068] During illumination, the samples were protected with aluminum foil to avoid any external influences that may occur.

[0069] After incubating the samples at 37°C for 24 hours, the number of bacterial colonies in the solid BHI medium Petri dishes was visually observed. Each experiment was performed in triplicate, and 3 to 30 colonies were counted. The CFU / mL for each set was calculated using the following formula: CFU / mL = (# colonies x # dilution) / volume The number of colonies is the average of the colony counts measured in triplicate for each experimental set.

[0070] All tubes were washed with PBS to remove planktonic cells, and then divided into four experimental groups. Biofilms formed on the surface of endotracheal tubes (ET). Biofilms formed on the surface of curcumin-functionalized tubes (MP1-PS1). 50 J / cm 2 Biofilm formed on the surface of ET irradiated with a light dose of 50 J / cm 2 Biofilm formed on the surface of MP1-PS1 irradiated with a light dose of 1000 kJ / L. Bacterial biofilm was removed from the tube surface with PBS. To evaluate the antibacterial activity of the groups, bacterial colony counts were plated on solid agar.

[0071] Therefore, regardless of photodynamics, this example demonstrates that curcumin-functionalized endotracheal tubes (ETs) using MP1-PS1 materials can inactivate microorganisms and prevent biofilm formation, and are therefore applicable to patients who are bedridden in hospital beds and require mechanical ventilation.

[0072] In a preferred embodiment of the present invention, the curcumin-functionalized endotracheal tube (ET) using the MP1-PS1 material has the ability to reduce microbial adhesion and promote microbial inactivation at a wavelength light source (450 nm) that is advantageous for the catheter decontamination process.

Claims

1. 1. A method for obtaining a functional polymer surface, said method comprising: a) dissolving a photosensitizer PS1 or PS3 of the following formula together with a base in an organic solvent selected from dimethylformamide (DMF), dimethoxyethane (DME), tetrahydrofuran (THF), dimethylpyrrolidone, dichloromethane, ethyl acetate, or dimethyl sulfoxide (DMSO); 【Chemistry 1】 b) immersing a polymer or copolymer material MP1 or MP2 of the formula shown below in the solution obtained in step (a) at a temperature between 0°C and 40°C; 【Chemistry 2】 c) subjecting the leaving group (X) of Cl of the polymer material MP1 or MP2 and the nucleophilic group (Y) selected from OH and N- present in the photosensitizer molecule PS1 or PS3 to a nucleophilic substitution reaction in a selected organic solvent, the duration of which is 0.5 to 48 hours; d) drying the surface functional polymer (MPn-PSm) at room temperature and under vacuum.

2. The base in step (a) is selected from triethylamine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), 1,5-diazabicyclo[4.3.0]non-5-ene (DBN), pyridine, or CaCO ３ , NaOH, KOH, Ba(OH) ２ , Al(OH) ３ , Mg(OH) ２ , Be(OH) ２ , Ca(OH) ２ The method of claim 1, wherein the compound is selected from the group consisting of:

3. 2. The method of claim 1, wherein the organic solvent in step (a) is dimethyl sulfoxide (DMSO).

4. 10. The method of claim 1, wherein step (c) is followed by a supplemental wash with ethanol.

5. The functional polymer material obtained by the method of any one of claims 1 to 4 has at least one polymer surface (MP1 or MP2), characterized in that a halogen leaving group on the surface is bound to a photosensitizer molecule (PS1 or PS3) by a stable chemical bond (D) of the ether type (O) or amine (NH).

6. The functional polymer material according to claim 5, wherein the polymer surface is MP1 or MP2, and the photosensitizer molecule is PS1.

7. The functional polymer material according to claim 5 is characterized in that the photosensitizer molecule PS1 is (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione (curcumin).

8. The functional polymer material described in claim 5 is characterized in that the photosensitizer molecule PS3 is a porphyrin functionalized with an amine-type nucleophilic group, or its reduced derivative, a chlorin or bacteriochlorin.

9. The functional polymer material according to claim 5 is characterized by the fact that the photosensitizer molecules of curcumin type (PS1) and tetrapyrrole macrocycle type (PS3) have a nucleophilic group (Y), said nucleophilic group being OH or N-.

10. The functional polymer material according to claim 5 exhibits antibacterial activity in the absence of light, and exhibits a more pronounced effect in the presence of light of a specific wavelength of 400 to 850 nm.

11. Use of the functional polymer material according to any one of claims 5 to 10 for the preparation, manufacture of polymeric biomedical devices.

12. 12. The use of claim 11, wherein the polymeric biomedical device is a medical device selected from an endotracheal tube, a probe, a catheter, a reservoir, a tracheostomy cannula, an intravenous infusion scalpel, a nasal oxygen catheter, a hemodialysis catheter, a rectal probe, an organ transport storage package, a urethral probe, or a tracheal suction probe.

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