Polymer including pentafluorophenyl ester and electrochemical biosensor including the same

Abstract

The present invention relates to poly(pentafluorophenyl ester)-based polymers for manufacturing electrochemical sensors, wherein the polymers have high reactivity with amine functional groups, cause less hydrolysis, maintain good solubility in organic solvents for general purposes, and have less steric hindrance and toxicity or side effects compared to polymers having similar structures, and thus are useful in insertion-type devices, especially continuous glucose monitoring systems whose part is inserted into the human body.

Description

Technical Field

[0001] Cross-reference to related applications

[0002] This application claims priority to Korean Patent Application No. 10-2020-0189140, filed on December 31, 2020, and all disclosures in the corresponding Korean patent application are incorporated herein by reference as a part thereof.

[0003] The present invention relates to polymers including pentafluorophenyl esters and electrochemical biosensors including such polymers, which can be used to immobilize electron transfer mediators and oxidoreductases on electrode surfaces in devices such as electrochemical biosensors.

[0004] Diabetes is a disease that occurs when high blood sugar levels are maintained for a long period and can lead to complications such as cardiovascular disease, stroke, and kidney disease. In response to high blood sugar levels, insulin injections are needed to lower them; however, if an excessive amount of insulin is injected, hypoglycemia can occur, leading to shock or death. To prevent this, it is essential for people with diabetes to continuously measure their blood glucose levels using a glucose sensor to maintain appropriate blood sugar levels.

[0005] Recently, a blood glucose sensor utilizing a continuous glucose monitoring system (CGMS) has been researched and commercialized. A CGMS sensor is a device inserted into subcutaneous tissue that continuously measures glucose concentration via intercellular fluid rather than blood. With finger-prick blood sampling, it is difficult to accurately monitor changes in blood glucose levels by measuring blood glucose 5 to 6 times daily, while CGMS has the advantage of monitoring changes and trends in blood glucose levels throughout the day. This allows patients to quickly identify hyperglycemia or hypoglycemia, and further improves their ability to manage blood glucose.

[0006] Biosensors are devices that selectively detect biological samples and convert them into specific signals. Enzyme-based biosensors using electrochemical methods are particularly preferred due to their improved selectivity, miniaturization, and measurement accuracy. Blood glucose biosensors using electrochemical methods are mainly divided into first-generation and second-generation methods. First-generation blood glucose sensors, initially developed by Clark and Lyon, measure blood glucose levels by detecting a decrease in oxygen concentration and a change in hydrogen peroxide concentration generated by an enzyme's redox reaction. Second-generation blood glucose sensors utilize electron transfer mediators to transfer electrons generated by the enzyme's redox reaction to electrodes. Compared to first-generation sensors, second-generation sensors offer several advantages: smaller errors dependent on oxygen concentration and more efficient and rapid electron transfer reactions via mediators. Therefore, second-generation sensor methods incorporating electron transfer mediators are being applied to CGMS blood glucose sensors.

[0007] Enzymatic blood glucose biosensors using electrochemical methods typically consist of an enzyme, an electron transfer mediator, and an electrode. First, glucose is oxidized by the enzyme to gluconolactone, and then the reduced enzyme is oxidized again and donates electrons to the electron transfer mediator. The reduced mediator is then oxidized and undergoes a process that transfers electrons to the electrode. From this series of processes, blood glucose levels can be identified as an electrical signal.

[0008] In the production of blood glucose sensors, methods for immobilizing electron transfer mediators and enzymes on electrode surfaces are crucial. To date, methods have been investigated for immobilizing redox hydrated polymers containing electron transfer mediators onto electrodes by mixing with enzymes and adding cross-linking agents. Polyvinylpyridine and polyvinylimidazole are well-known redox polymer matrices. However, these conventional redox polymers suffer from long and complex synthesis steps, exhibit low immobilization efficiency of transition metal complexes, and difficulty in incorporating other functional groups into the polymer. Therefore, even now, to develop redox polymers with superior performance, new materials that transcend the limitations of conventional materials are needed.

[0009] Against this backdrop, the inventors repeatedly studied polymers for immobilizing electron transfer mediators and enzymes for electrochemical biosensors. As a result, they demonstrated that when using polymers containing pentafluorophenyl (PFP) esters, they exhibit high reactivity with amine functional groups, less hydrolysis, and generally good solubility in organic solvents. Furthermore, they showed less steric hindrance and toxicity compared to polymers with similar structures. In particular, they have demonstrated that this is useful for implantable devices (e.g., continuous glucose monitoring sensors) in which a portion of the sensor is inserted into the human body, thus completing the present invention. Summary of the Invention

[0010] [Technical Issues]

[0011] The object of this invention is to provide a polymer comprising pentafluorophenyl ester for manufacturing redox polymer materials.

[0012] Another object of the present invention is to provide redox polymer material thin films and electrochemical biosensors comprising polymers used in the manufacture of transition metal complexes and redox polymer materials.

[0013] [Technical Solution]

[0014] As one aspect of achieving the above objectives, the present invention relates to polymers comprising pentafluorophenyl esters for manufacturing redox polymer materials, redox polymers comprising the polymer and transition metal complexes for electrochemical biosensors, redox polymer material films comprising redox polymers for electrochemical biosensors prepared therefrom, and electrochemical biosensors comprising the same, such as blood glucose sensors.

[0015] [Beneficial Effects]

[0016] When used in electrochemical sensors, the polypentafluorophenyl ester polymers according to the invention have high reactivity with amine functional groups, less hydrolysis, high solubility in organic solvents, and low steric hindrance and toxicity or side effects, and therefore, in particular, can be used in implantable devices in which a portion of the sensor is placed inside the human body, such as continuous glucose monitoring sensors. Attached Figure Description

[0017] Figure 1 To illustrate the effect of adding osmium compound 22 to the PFP polymer P1 according to the present invention, 19 A graph showing the reaction monitoring results of F NMR.

[0018] Figure 2 shows the confirmation of the final reaction of the PFP polymer (36) and the osmium complex (22, 33). 19 The F NMR curve, and the proportion of redox polymers (37, 38) predicted by the curve.

[0019] Figure 3 This confirms the synthesis of redox polymers comprising transition metal complexes containing CN ligands and PFP polymers as described in Examples 1-4. 19 F NMR data.

[0020] Figure 4 To confirm the synthesis of the redox polymers in Examples 1-5, which include transition metal complexes containing CN ligands and PFP polymers. 19 F NMR data.

[0021] Figure 5 This is a schematic diagram of a composition of redox polymers and enzymes, and a crosslinking method on the surface of SPCE electrodes.

[0022] Figure 6aTo illustrate the changes in current (upper part) as the glucose concentration in the redox polymers (37 and 38) according to the invention increases to 0.01, 0.05, 0.1, 0.5, 1, 2, 4, 8, 16, 32, 64, and 100 mM per 200 seconds, the graphs are shown below, with the left side representing the low concentration portion (0–1 mM) and the right side representing the total portion (0–100 mM).

[0023] Figure 6b The graphs illustrate the changes in current (top left) as the glucose concentration in the redox polymer (including redox polymer 1 containing a transition metal complex with a CN ligand and a PFP polymer) increases to 0.01, 0.05, 0.1, and 0.5 mM per 200 seconds, the changes in current according to glucose concentration (top right), and the changes in current (bottom left) as the glucose concentration increases to 0, 1, 5, 10, 50, and 100 mM per 200 seconds.

[0024] Figure 6c The graphs illustrate the changes in current (top left) as the glucose concentration in the redox polymer (including redox polymer 1 containing a transition metal complex with a CN ligand and a PFP polymer) increases to 0.01, 0.05, 0.1, and 0.5 mM per 200 seconds, the changes in current according to glucose concentration (top right), and the changes in current (bottom left) as the glucose concentration increases to 0, 1, 5, 10, 50, and 100 mM per 200 seconds.

[0025] Figures 7a to 7d A graph illustrating the measurement results of the current changes before and after cyclic voltammetry testing of the redox polymers (37, 38 and redox polymers 1 and 2, including transition metal complexes containing CN ligands and PFP polymers) according to the present invention. Figures 7a to 7d In this context, CV represents the value measured immediately after electrode completion, and the value following CV represents the value measured after stirring in PBS solution for 1 hour. Detailed Implementation

[0026] The invention will now be described in more detail.

[0027] The polymers used in the manufacture of redox polymer materials according to the present invention are polymers based on polypentafluorophenyl esters, and specifically, polymer precursors used as redox polymer materials include repeating units derived from pentafluorophenyl esters. Preferably, the polymers can be crosslinked with reactive groups including amine groups and transition metal complexes to form redox polymer materials for electron transfer mediators.

[0028] As an example, the polymer for manufacturing redox polymer materials according to the present invention may be a polymer for manufacturing redox polymer materials, which is prepared by introducing primary and secondary amine compounds, which are crosslinking materials containing amine groups, into polypentafluorophenyl ester via ammonolysis.

[0029] Non-limiting examples of polymers used in redox polymer materials may be one or more polymers selected from the group consisting of: polypentafluorophenyl acrylate (PPFPA) homopolymer, polypentafluorophenyl methacrylate (PPFPM) homopolymer, polypentafluorophenyl acrylate-polydimethacrylamide (PPFPA-PDMA) copolymer, polypentafluorophenyl methacrylate-polydimethacrylamide (PPFPM-PDMA) copolymer, polypentafluorophenyl acrylate-polyacrylamide (PPFPA-PAA) copolymer, polypentafluorophenyl methacrylate-polyacrylamide (PPFPM-PAA) copolymer, but are not limited thereto.

[0030] In one example, the polymer according to the invention for manufacturing redox polymer materials may have the structure of either Chemical Formula 1 or Chemical Formula 2:

[0031] [Chemical Formula 1]

[0032]

[0033] [Chemical Formula 2]

[0034]

[0035] In chemical formula 1 or chemical formula 2,

[0036] R T and R L Each is independently selected from the group consisting of: substituted or unsubstituted alkylene groups having 1 to 20 carbon atoms, substituted or unsubstituted cycloalkylene groups having 1 to 20 carbon atoms, substituted or unsubstituted glycol groups having 3 to 30 carbon atoms, substituted or unsubstituted aryl groups having 6 to 30 carbon atoms, substituted or unsubstituted heteroaryl groups having 3 to 30 carbon atoms, substituted or unsubstituted alkenyl groups having 2 to 40 carbon atoms, and substituted or unsubstituted alkynyl groups having 2 to 40 carbon atoms; and

[0037] n is an integer from 10 to 300.

[0038] In this invention, "substitution" can be at least one hydrogen atom selected from one to three of the following groups: halogen atom (e.g., F, Cl, Br or I), cyano, hydroxyl, thiol, nitro, amino, imino, azide, amidine, hydrazine, hydrazone, oxo, carbonyl, carbamoyl, ester, ether, carboxylic acid or its salt, sulfonic acid or its salt, phosphate or its salt, alkyl having 1-6 carbon atoms, haloalkyl having 1-6 carbon atoms, alkenyl having 2 to 6 carbon atoms, haloalkenyl having 2 to 6 carbon atoms, and so on. Alkynyl groups having 6 carbon atoms, haloalkynyl groups having 2 to 6 carbon atoms, alkoxy groups having 1 to 6 carbon atoms, haloalkoxy groups having 1 to 6 carbon atoms, alkylthio groups having 1 to 6 carbon atoms, heterocyclic alkyl groups having 1 to 9 carbon rings, aryl groups having 6 to 10 carbon atoms, aryloxy groups having 6 to 10 carbon atoms, arylthiol groups having 6 to 10 carbon atoms, heteroalkyl groups having 1 to 9 carbon rings, heteroaryloxy groups having 1 to 9 carbon rings, and heteroarylthio groups having carbon rings containing 1 to 9 carbon atoms, unless otherwise mentioned.

[0039] Preferably, R T and R L Each of the following can be independently selected: polydimethylacrylamide (PDMA), polyacrylamide (PAA), polystyrene (PS), polyethylene glycol (PEG), polyethylene oxide (PEO), polymethyl methacrylate (PMMA), polyvinyl imidazole (PVI), polyvinylpyridine (PVP), and polydimethoxysiloxane (PDMS) with a molecular weight of 1,000 g / mol to 50,000 g / mol.

[0040] As an example, the polymers used in the manufacture of redox polymer materials according to the present invention may have a weight-average molecular weight in the range of 1,000 g / mol to 500,000 g / mol, but are not limited thereto.

[0041] In another aspect, the present invention relates to redox polymer materials for electrochemical sensors, wherein a transition metal complex is incorporated into a polypentafluorophenyl ester polymer.

[0042] For example, the redox polymer material can be prepared by introducing a compound comprising a functional group selected from the group consisting of amino, ammonium, halogen, epoxy, azide, acrylate, alkenyl, alkynyl, thiol, isocyanate, alcohol and silyl groups to functionalize the polypentafluorophenyl ester polymer, and incorporating a transition metal complex into the thus functionalized polymer.

[0043] Alternatively, redox polymer materials can be prepared by introducing functional groups, such as crosslinking materials (e.g., amine, ammonium, thiol, and alcohol groups), into the ligands of the transition metal complex via substitution and addition reactions, and then combining the so-functionalized transition metal complex with the polymer according to the invention, thereby functionalizing the polypentafluorophenyl ester polymer rather than functionalizing the polymer.

[0044] Alternatively, polymeric materials can be prepared by functionalizing all polypentafluorophenyl ester polymers and transition metal complexes using crosslinking materials containing functional groups selected from the group consisting of amino, ammonium, halogen, epoxy, azide, acrylate, alkenyl, alkynyl, thiol, isocyanate, alcohol, and silane, and then bonding them together.

[0045] Preferably, in the redox polymer material according to the invention, a functional group and a transition metal complex selected from the group consisting of primary amines, secondary amines and ammonium groups are incorporated into the polypentafluorophenyl ester polymer.

[0046] Specifically, transition metal complexes may have the following structures: chemical formula 3 or chemical formula 4.

[0047] [Chemical Formula 3]

[0048]

[0049] [Chemical Formula 4]

[0050]

[0051] In chemical formula 3 or chemical formula 4

[0052] M is a transition metal selected from the group consisting of: Os, Rh, Ru, Ir, Fe, and Co; and

[0053] In chemical formula 3, L G1 and L G2 They combine with each other to form bidentate ligands selected from chemical formulas 5 to 6 below; and

[0054] L G3 and L G4 They combine to form bidentate ligands selected from chemical formulas 5 to 6 below; and L G5 and L G6 They combine with each other to form bidentate ligands selected from chemical formulas 5 to 6 below; and

[0055] [Chemical Formula 5]

[0056]

[0057] [Chemical Formula 6]

[0058]

[0059] In chemical formula 5, L C It is a heterocyclic compound comprising one or more nitrogen atoms, and is connected to benzene at the 2-position; and

[0060] In chemical formula 6, L N For heterocyclic compounds comprising one or more nitrogen atoms, and L N1 and L N2 They are connected to each other in two positions.

[0061] R L L is a heterocyclic compound C L N1 and L N2 All functional groups.

[0062] In one aspect, R1, R2, R3, R4, R5 and R L Each can be independently selected from the group consisting of: substituted or unsubstituted alkyl groups having 1 to 10 carbon atoms, substituted or unsubstituted glycol groups having 2 to 20 carbon atoms, substituted or unsubstituted alcohol groups having 1 to 20 carbon atoms, substituted or unsubstituted haloalkyl groups having 1 to 20 carbon atoms, substituted or unsubstituted thiol groups having 1 to 20 carbon atoms, substituted or unsubstituted alkyl azide groups having 3 to 20 carbon atoms, substituted or unsubstituted aryl azide groups having 7 to 30 carbon atoms, substituted or unsubstituted alkenyl groups having 2 to 40 carbon atoms, substituted or unsubstituted alkynyl groups having 2 to 40 carbon atoms, cyano groups, halogen groups, deuterium, and hydrogen.

[0063] In one aspect, L of chemical formula 4 H1 L H3 and L H4 L H2 They bond together around a central point to form a tetradentate ligand represented by the following chemical formula 7.

[0064] Then, L H5 and L H6 They can respectively form monodentate ligands of transition metal M represented by chemical formula 8; or L H5 and L H6 They can combine with each other to form bidentate ligands of transition metal M, as represented by the following chemical formula 9; and

[0065] [Chemical Formula 7]

[0066]

[0067] In chemical formula 7,

[0068] C a C b and C c It is a heterocyclic compound comprising one or more nitrogen atoms, and preferably, the heterocyclic compound may be linked to methylamine at its 2 position, and the three nitrogen atoms of the three heterocycles and the one nitrogen atom at the center connecting the three heterocycles may be linked to the transition metal M.

[0069] R1, R2, and R3 refer to heterocyclic compounds C a C b C c All functional groups.

[0070] In one aspect, R1, R2, and R3 may each be independently selected from the group consisting of: substituted or unsubstituted alkyl groups having 1 to 10 carbon atoms, substituted or unsubstituted glycol groups having 2 to 20 carbon atoms, substituted or unsubstituted alcohol groups having 1 to 20 carbon atoms, substituted or unsubstituted haloalkyl groups having 1 to 20 carbon atoms, substituted or unsubstituted thiol groups having 1 to 20 carbon atoms, substituted or unsubstituted alkyl azide groups having 3 to 20 carbon atoms, substituted or unsubstituted aryl azide groups having 7 to 30 carbon atoms, substituted or unsubstituted alkenyl groups having 2 to 40 carbon atoms, substituted or unsubstituted alkynyl groups having 2 to 40 carbon atoms, cyano groups, halogen groups, deuterium groups, and hydrogen groups. Preferably, each may independently introduce a reactive group capable of being attached to the polymer.

[0071] [Chemical Formula 8]

[0072]

[0073] [Chemical Formula 9]

[0074]

[0075] In chemical formula 8,

[0076] L H5 and L H6 It can be a monodentate ligand independently, and each can be a heterocyclic compound comprising one or more -H, -F, -Cl, -Br, -I, -NO2, -NCCH3, -CO, -OH2, -NH3 or nitrogen atoms.

[0077] In chemical formula 9,

[0078] L H5 -L H6 It can be a bidentate ligand, and can be catechol, acetylacetone, 2-pyridinecarboxylic acid, 2-pyridinecarboxamide, 2,2-bipyridine, or 2,2-bithiazole.

[0079] L1 can be independently selected from the group consisting of: substituted or unsubstituted alkylene groups having 1 to 20 carbon atoms, substituted or unsubstituted cycloalkylene groups having 1 to 20 carbon atoms, substituted or unsubstituted glycol groups having 2 to 30 carbon atoms, substituted or unsubstituted arylene groups having 6 to 30 carbon atoms, and substituted or unsubstituted heteroarylene groups having 3 to 30 carbon atoms; and

[0080] A a Select from the group consisting of amino and ammonium groups.

[0081] One aspect of the present invention relates to a method for preparing a redox polymer material thin film, comprising coating a redox polymer material comprising a polymer onto an electrode and then curing the coated electrode; and the redox polymer material thin film thus prepared and an electrochemical biosensor comprising the thin film.

[0082] Additionally, another aspect of the invention relates to a sensing layer for an electrochemical biosensor, comprising a C enzyme capable of oxidizing and reducing liquid biological samples; and an electron transfer mediator comprising a transition metal complex.

[0083] Oxidoreductases are enzymes that catalyze redox reactions in living organisms, and in the context of this invention, in the case of a target material (e.g., a biosensor), they refer to enzymes that are reduced by reacting with the target material. The reduced enzyme then reacts with an electron transfer mediator, and the target material is then quantified by measuring a signal (e.g., a change in the generated current). The oxidoreductases available in this invention can be one or more selected from the group consisting of various dehydrogenases, oxidases, and esterases, and can be used by selecting enzymes from the group of enzymes that use the target material as a substrate, based on whether redox reactions are occurring or the target material is being detected.

[0084] More specifically, oxidoreductases may be one or more selected from the group consisting of: glucose dehydrogenase, glutamate dehydrogenase, glucose oxidase, cholesterol oxidase, cholesterol esterase, lactate oxidase, ascorbic acid oxidase, alcohol oxidase, alcohol dehydrogenase, and bilirubin oxidase, etc.

[0085] On the other hand, the oxidoreductase may include a cofactor that serves to store the hydrogen stolen by the oxidoreductase from the target material (e.g., the target material), and for example, it may be one or more selected from the group consisting of: flavin adenine dinucleotide (FAD), nicotinamide adenine dinucleotide (NAD), and pyrroloquinoline quinone (PQQ), etc.

[0086] For example, when measuring blood glucose concentration, glucose dehydrogenase (GDH) can be used as an oxidoreductase, and glucose dehydrogenase can be flavin adenine dinucleotide-glucose dehydrogenase (FAD-GDH) with FAD as a cofactor, and / or nicotinamide adenine dinucleotide-glucose dehydrogenase with FAD-GDH as a cofactor.

[0087] In specific embodiments, the available oxidoreductases may be one or more selected from the group consisting of: FAD-GDH (e.g., EC 1.1.99.10, etc.), NAD-GDH (e.g., EC 1.1.1.47, etc.), PQQ-GDH (e.g., EC 1.1.5.2, etc.), glutamate dehydrogenase (e.g., EC 1.4.1.2, etc.), glucose oxidase (e.g., EC 1.1.3.4, etc.), cholesterol oxidase (e.g., EC 1.1.3.6, etc.), cholesterol esterase (e.g., EC 3.1.1.13, etc.), lactate oxidase (e.g., EC 1.1.3.2, etc.), ascorbic acid oxidase (e.g., EC 1.10.3.3, etc.), alcohol oxidase (e.g., EC 1.1.3.13, etc.), alcohol dehydrogenase (e.g., EC 1.1.1.1, etc.), and bilirubin oxidase (e.g., EC 1.3.3.5, etc.), etc.

[0088] Most preferably, the oxidoreductase is a glucose dehydrogenase that can maintain 70% or higher activity for one week in a buffer solution at 37°C.

[0089] Based on 100 parts by weight of oxidoreductase, the sensing layer according to the present invention may contain 20 to 700 parts by weight, for example, 60 to 700 parts by weight, or 30 to 340 parts by weight of oxidoreductase. The content of oxidoreductase may be appropriately adjusted according to the activity of oxidoreductase.

[0090] Furthermore, the sensing layer according to the invention may further include carbon nanotubes for enhancing membrane performance. Specifically, when used with transition metal composites (particularly osmium), the carbon nanotubes can further enhance the performance of the sensing layer as the electron transfer rate increases.

[0091] In addition, the sensing layer according to the present invention may further include a crosslinking agent.

[0092] On the other hand, the sensing layer according to the invention may further include one or more additives selected from the group consisting of surfactants, aqueous polymers, tertiary ammonium salts, fatty acids and thickeners, etc., for acting as a dispersant during reagent dissolution, as a binder during reagent preparation, and as a stabilizer for long-term storage, etc.

[0093] When the composition is aliquoted, the surfactant can serve to uniformly distribute the composition on the electrode to achieve a uniform thickness. As a surfactant, one or more selected from the group consisting of Triton X-100, sodium dodecyl sulfate, perfluorooctane sulfonate, and sodium stearate can be used. Based on 100 parts by weight of oxidoreductase, the reagent composition according to the invention may contain 3 to 5 parts by weight, for example, 10 to 25 parts by weight of surfactant, so as to suitably achieve a uniform distribution of the reagent on the electrode to achieve a uniform thickness when the reagent is aliquoted. For example, when using an oxidoreductase with an activity of 700 U / mg, based on 100 parts by weight of oxidoreductase, 10 to 25 parts by weight of surfactant may be included, and when the activity of the oxidoreductase is higher than 700 U / mg, the surfactant content can be adjusted to a lower content than the above.

[0094] The aqueous polymer serves as the polymer carrier for the reagent composition, aiding in the stability and dispersion of the enzyme. As the aqueous polymer, one or more selected from the group consisting of: polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), polyperfluorosulfonate, hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), carboxymethyl cellulose (CMC), cellulose acetate, and polyamide, etc. Based on 100 parts by weight of the oxidoreductase, the reagent composition according to the invention may contain 10 to 70 parts by weight, for example, 30 to 70 parts by weight of the aqueous polymer, to adequately and appropriately demonstrate its role in stabilizing and dispersing the oxidoreductase. For example, when using an oxidoreductase with an activity of 700 U / mg, based on 100 parts by weight of the oxidoreductase, 30 to 70 parts by weight of the aqueous polymer may be contained, and when the activity of the oxidoreductase is higher than 700 U / mg, the content of the aqueous polymer can be adjusted to a content lower than the above.

[0095] Aqueous polymers may have a weight-average molecular weight of about 2,500 g / mol to 3,000,000 g / mol, for example, 5,000 g / mol to 1,000,000 g / mol, in order to effectively help stabilize and disperse the carrier and enzyme.

[0096] The thickener serves to firmly adhere the reagent to the electrode. One or more substances selected from natrosol and diethylaminoethyl dextran hydrochloride (DEAE-dextran hydrochloride) can be used as the thickener. Based on 100 parts by weight of the oxidoreductase, the electrochemical sensor according to the invention may contain 10 to 90 parts by weight, for example 30 to 90 parts by weight, of the thickener to firmly attach the redox polymer according to the invention to the electrode. For example, when using an oxidoreductase with an activity of 700 U / mg, based on 100 parts by weight of the oxidoreductase, 30 to 90 parts by weight of the thickener may be included, and when the activity of the oxidoreductase is higher than 700 U / mg, the content of the thickener can be adjusted to a content lower than the above.

[0097] In another aspect, the present invention can be a device, preferably an insertable device, for example, an insertable device for the human body, comprising the organic electron transfer mediator. Additionally, preferably, the device can be an electrochemical biosensor, more preferably, an electrochemical glucose (blood sugar) sensor.

[0098] Specifically, there are no restrictions on the type of electrochemical biosensor, but it can be a continuous blood glucose monitoring sensor.

[0099] As components of this continuous glucose monitoring sensor, the present invention may include, for example, electrodes, insulators, substrates, a sensing layer comprising a redox polymer and a redox enzyme, a diffusion layer, and a protective layer. In the case of electrodes, two electrodes may be included, such as a working electrode and a counter electrode, and three electrodes may be included, such as a working electrode, a counter electrode, and a reference electrode. In one embodiment, the biosensor according to the present invention may be an electrochemical biosensor, which is generated by applying a reagent composition comprising a redox polymer to a substrate having at least two, preferably two or three electrodes and drying it. The redox polymer comprises an organic series of electron transfer mediators of Formula 1 and an enzyme capable of redox-reducing liquid biological samples. For example, in the electrochemical biosensor, a planar electrochemical biosensor is provided, wherein the working electrode and the counter electrode are mounted on opposite sides of each other, and a sensing layer comprising a redox polymer having an organic series of electron transfer mediators according to the present invention is laminated on the working electrode, and an insulator, a diffusion layer, and a protective layer are sequentially stacked on both sides of the substrate equipped with the working electrode and the counter electrode.

[0100] In a specific aspect, the substrate may be made of one or more materials selected from the group consisting of PET (polyethylene terephthalate), PC (polycarbonate), and PI (polyimide).

[0101] Alternatively, the working electrode can be a carbon, gold, platinum, silver, or silver / silver chloride electrode.

[0102] Furthermore, in the case of an electrochemical biosensor with two electrodes, the counter electrode also serves as a reference electrode. Therefore, gold, platinum, silver, or silver / silver chloride electrodes can be used as the counter electrode. In the case of an electrochemical biosensor with three electrodes including a reference electrode, gold, platinum, silver, or silver / silver chloride electrodes can be used as the reference electrode, and a carbon electrode can be used as the counter electrode.

[0103] Nafion, cellulose acetate, and silicone rubber can be used as the diffusion layer, and silicone rubber, polyurethane, and polyurethane copolymers can be used as the protective layer, but are not limited to these.

[0104] As a non-limiting example, in the case of two electrodes, the counter electrode also serves as a reference electrode, so silver chloride or silver can be used. In the case of three electrodes, silver chloride or silver can be used as the reference electrode, and a carbon electrode can be used as the counter electrode.

[0105] The specific embodiments of the present invention illustrate a biosensor for measuring glucose as an example of an applicable electrochemical biosensor, but by changing the type of enzyme contained in the reagent composition of the present invention, it can be applied to biosensors for quantifying various substances (such as cholesterol, lactate, creatinine, hydrogen peroxide, alcohols, amino acids, and glutamate).

[0106] [Invention Method]

[0107] The present invention will be described in more detail below through the following embodiments. However, the following embodiments are merely illustrative of the invention, and the scope of the invention is not limited to the following embodiments.

[0108] [Example]

[0109] Experimental materials

[0110] Refer to Example 1. Reagents and Samples

[0111] Reagents commercially available from Aldrich, Acros, TCI, and Alfa Aesar were used without special purification. The solution was obtained by filling 10 mL pipettes with basic alumina and then filtering, using ethylenediamine, pyridine, 1-methylimidazole, and triethylamine. Purification was performed using solvents purchased from Daejung and Samchun. Anhydrous dichloromethane and anhydrous N,N-dimethylformamide were obtained by adding calcium hydride and distillation, and anhydrous acetonitrile was obtained by distillation using P2O5. Diethyl ether was distilled using sodium and benzophenone. For thin-layer chromatography used in the analysis, Merck silica gel 60F was used. 254A glass plate was used, and fluorescence was observed using dual short-wavelength (254 nm) / long-wavelength (365 nm) UV lamps. For column chromatography, Aldrich basic alumina or Alfa neutral alumina were used. Zensor SE102 was used as the screen-printed carbon electrode (SPCE).

[0112] Refer to Example 2. Device

[0113] Using Varian Inova 400 (for 1 H is 400MHz, and for 13 A spectrometer with a C value of 100 MHz obtained hydrogen nuclear magnetic resonance (NMR) 1 H NMR), carbon nuclear magnetic resonance (H NMR), 13 C NMR and fluorine nuclear magnetic resonance (C NMR) 19 F NMR) spectrum. Based on tetramethylsilane (δ 0.00) or deuterated chloroform (F NMR) 1 The δ of CDCl3 in H NMR is 7.26. 13 In C NMR, CDCl3 is δ 77.2, and deuterated dimethyl sulfoxide (DMS) is δ 77.2. 1 In H NMR, the value for DMSO is δ2.50. 13 In C NMR, δ for DMSO is 39.52 and for deuterated acetonitrile ( 1 In 1H NMR, the value for CD3CN is δ 1.94. All chemical shifts are expressed in ppm. In fluorine NMR (… 19 In the case of F NMR), a small amount of trifluorotoluene (CF3-Ar, 19 In F NMR, CD3OD (δ -69.27) was added to deuterated methanol and the results are expressed in ppm. Cyclic voltammetry (CV) measurements were performed using a CHI1040C model from CH Instruments. Measurements were taken at scan rates of 10–50 mV / s using a cleaned 3 mm diameter carbon glass electrode as the working electrode, an Ag / AgCl electrode as the reference electrode, and a Pt electrode as the counter electrode. Mass spectra were obtained at low resolution using an ESI-Iontrap model from Thermo Fisher Scientific at the Organic Chemistry Research Center, Sogang University, and at high resolution using an ESI-orbitrap model. Single-crystal X-ray diffraction analysis was performed by the Korea Institute of Chemical Technology, and the crystal structure was identified using a Bruker SMART APEX II model.

[0114] Example 1. Synthesis of redox polymers using a PFP polymer matrix

[0115] The aim of this experiment was to synthesize redox polymers from a pentafluorophenyl acrylate (PFPA) matrix via reaction with an electron transfer mediator containing an amine group (-NH2) at the end. First, using AIBN, a DMA:PFPA polymer (P1) was synthesized by free radical polymerization of pentafluorophenyl acrylate (PFPA) and dimethacrylamide (DMA) in a 5:5 ratio.

[0116] 1-1. Synthesis of poly(N,N-dimethylacrylamide-co-pentafluorophenyl acrylate) (P1)

[0117]

[0118] [Synthesis of DMA:PFP polymers]

[0119] N,N-dimethylacrylamide (150 mg, 1.51 mmol), pentafluorophenyl acrylate (360 mg, 1.51 mmol), and azobisisobutyronitrile (AIBN, 7 mg, 0.043 mmol) were added to a glass culture tube, and acetonitrile (8 mL) was added to prepare a mixed solution. Argon gas was then purged into the glass culture tube for 10 minutes to create an argon atmosphere, and the mixture was refluxed at 80 °C for 12 hours. After the reaction was complete, the solvent was removed using a rotary evaporator, and CH₂Cl₂ was added to prepare a mixed solution. The polymer obtained by precipitating the mixture three times by slowly adding the mixed solution dropwise to n-hexane was purified. The polymer obtained by drying the filtered precipitate and washing it under vacuum with n-hexane for 6 hours or longer yielded product P1 as a white solid (482 mg, 95%). 1 ¹H NMR (400MHz, CDCl₃) δ 2.81–3.34 ppm (br, 7H, -CH- in the PPFPA skeleton and -N(CH₃)₂ from PDMA), 2.47–2.65 ppm (br, 1H, -CH- in the PDMA skeleton), 1.13–2.22 ppm (br, 4H, -CH₂- in the PDMA and PPFPA skeletons). 19 F NMR (400MHz, CDCl3) δ -152.90 (br, ortho), -157.21 (br, para), -162.15 (br, meta)

[0120] The synthesized polymer exhibits good solubility in methanol, acetonitrile, acetone and dichloromethane, but is completely insoluble in water and diethyl ether.

[0121] 1-2. Confirm whether PFP polymers can be used as polymer matrices capable of immobilizing electron transfer mediators and GDH.

[0122] To confirm whether the reaction between the polymer P1 synthesized in Examples 1-1 and the osmium complex 22 comprising amine groups at the ends proceeded, by... 19Observations were made using F NMR.

[0123]

[0124] Complex 22 has a zwitterion Cl- as its zwitterion. - The reaction was carried out in the form of [missing information], and NMR peaks were confirmed in a deuterated methanol solvent capable of dissolving the complex and polymer. Additionally, when the amino group of complex 22 is in the form of ammonium (-NH4) [missing information], [missing information]. + When present in its original form, it cannot react with PFP ester, so triethylamine (TEA) in an equal amount to the complex was added. The results are as follows... Figure 1 As shown in the image. (If it can be obtained from...) Figure 1 It was confirmed that initially, through 19 F NMR confirmed only polymer P1, showing three broad peaks corresponding to PFPA. Then, upon the addition of complex 22, three additional peaks of pentafluorophenol (PFPOH) in a sharp form were observed. These are the peaks of PFPOH, a byproduct generated via the reaction of the amine at the end of complex 22. Over time, the PFPOH peaks gradually increased while the PFPA peaks gradually decreased; after approximately 36 hours, all PFPA peaks disappeared, leaving only the PFPOH peaks. These results confirm that the PFP ester can react with the amine group of a TPMA-like osmium complex, and that the PFPA polymer can be used as a polymer matrix capable of immobilizing electron transfer mediators and GDH.

[0125] 1-3. Synthesis of redox polymers (redox polymers (36) and (37))

[0126] Redox polymers were synthesized by reacting TPMA-based osmium complexes 22 and 33, which contain amine functional groups, with PFPA polymer 36 (Figure 2). To obtain redox polymers containing approximately 25%–30% osmium complexes throughout the polymer chain, the reaction was carried out by calculating the stoichiometric ratio.

[0127] 1) Synthesis of redox polymer (36)

[0128]

[0129] Starting material 22 (6 mg, 0.009 mmol) was dissolved in deuterated methanol (0.5 mL) in an NMR tube. The dissolved solution was added to starting material P1 (5 mg) dissolved in deuterated methanol (0.5 mL), and then triethylamine (1.24 μL, 0.009 mmol) was added. The NMR was then... 19After confirming the reaction progress by ¹H NMR, purification was achieved by repeatedly adding the reaction solution dropwise to diethyl ether three times to obtain a precipitate of the polymer. The resulting precipitate was filtered and washed with diethyl ether, then dried under vacuum for 4 hours or longer to obtain product 36 (6 mg, 64%) as a brown solid. 19 F NMR (400MHz, CD3OD) δ -153.38 (br, ortho), -159.74 (br, para), -163.53 (br, meta)

[0130] 2) Synthesis of redox polymer (37)

[0131]

[0132] Starting material 33 (6.5 mg, 0.009 mmol) was dissolved in deuterated methanol (0.5 mL) in an NMR tube. The dissolved solution was added to starting material P1 (5 mg) dissolved in deuterated methanol (0.5 mL), and then triethylamine (1.22 μL, 0.009 mmol) was added. 19 After confirming the reaction progress by ¹H NMR, purification was achieved by repeatedly adding the reaction solution dropwise to diethyl ether three times to obtain a precipitate of the polymer. The resulting precipitate was filtered and washed with diethyl ether, then dried under vacuum for 4 hours or longer to obtain product 37 (7.2 mg, 73%) as a brown solid. 19 F NMR (400MHz, CD3OD) δ -154.57 (br, ortho), -160.27 (br, para), -164.95 (br, meta)

[0133] pass 19 ¹H NMR confirmed the reaction between the added complex and the PFPA polymer until the peak no longer changed, and determined the redox polymer ratio. The reaction of all added osmium complexes took approximately 24–36 hours. As a result, [the reaction proceeded via]... 19 The FNMR results confirming the ratio of the two redox polymers indirectly determined that in redox polymer 37, the PFPA:PFPOH ratio was 2:3, with the osmium complex comprising 30% of the total chain, and in redox polymer 38, the PFPA:PFPOH ratio was 2.2:2.8, with 28% of the osmium complex contained in the polymer matrix. Both polymers were obtained by precipitation in diethyl ether, and unlike starting material polymer 36, all redox polymers were water-soluble. Furthermore, as confirmed below, the redox potential exhibited when measuring CV was similar to that of the osmium complex monomer. These results confirm the successful synthesis of a redox polymer containing a TPMA-like osmium complex.

[0134] 1-4. Synthesis of redox polymer 1, including a transition metal complex containing a CN ligand and a PFP polymer.

[0135]

[0136] As a starting material, an osmium complex [Os(2-(2-pyridyl-κN)-5-methaneamino-phenyl-κC)(4,4'-dimethyl-2,2'-bipyridine)2]Cl2 (11 mg, 0.014 mmol) including the CN ligand was dissolved in deuterated methanol (0.5 mL) in an NMR tube. The dissolved solution was added to a starting material PFP polymer (7:3) (17 mg) dissolved in deuterated methanol (0.5 mL), and then triethylamine (3.8 μL, 0.028 mmol) was added. The NMR was then... 19 F NMR ( Figure 3 After confirming the progress of the reaction, the polymer was purified by repeatedly adding the reaction solution dropwise to diethyl ether three times to obtain a precipitate. The resulting precipitate was filtered and washed with diethyl ether, then dried under vacuum for 4 hours or longer to obtain the product as a brown solid.

[0137] 1-5. Synthesis of redox polymers 2, including transition metal complexes containing CN ligands.

[0138]

[0139] In an NMR tube, the starting material [Os(2-(2-pyridyl-κN)-5-methaneamine-phenyl-κC)(4,4'-dimethoxy-2,2'-bipyridine)2]Cl2 (9.7 mg, 0.012 mmol) was dissolved in deuterated methanol (0.5 mL). The dissolved solution was added to the starting material PFP polymer (7:3) (15 mg) dissolved in deuterated methanol (0.5 mL), and then triethylamine (3.1 μL, 0.024 mmol) was added. The NMR was then... 19 After F NMR confirmed the progress of the reaction ( Figure 4 The polymer was purified by repeatedly adding the reaction solution dropwise to diethyl ether three times to obtain a precipitate. The resulting precipitate was filtered and washed with diethyl ether, then dried under vacuum for 4 hours or longer to obtain the product as a brown solid.

[0140] Example 2. Immobilization of redox polymers and enzymes and testing of their response to glucose.

[0141] A solution of a redox polymer containing an osmium complex and glucose dehydrogenase (GDH) was immobilized on SPCE electrodes, and the following tests were performed to confirm the response depending on the glucose concentration.

[0142] 2-1. Preparation of an electrode in which redox polymers and glucose dehydrogenase are immobilized

[0143] A mixed solution was prepared by mixing a solution containing 0.4 mg of a redox polymer dissolved in 50 μL of distilled water and a solution containing 0.8 mg of glucose dehydrogenase (GDH) dissolved in 50 μL of distilled water (v / v 1:1). 0.83 μL of this mixed solution in the cases of redox polymers 37 and 38, and 1 μL in the cases of redox polymers 1 and 2 containing transition metal complexes with CN ligands, were uniformly placed on SPCEs and slowly dried at room temperature. After drying, the 0.83 μL of the mixed solution in the cases of redox polymers 37 and 38, and 1 μL in the cases of redox polymers 1 and 2 containing transition metal complexes with CN ligands, were again placed on the SPCEs, and this process was repeated more than twice. Electrodes were prepared by drying the electrodes with a mixed solution containing a total of 2.5 μL (polymers 37 and 38) and 3 μL (including redox polymers 1 and 2 containing transition metal complexes with CN ligands) at room temperature for 12 hours or longer. Figure 5 The composition of the mediator and enzyme, as well as the crosslinking method on the surface of SPCE electrodes, are schematically shown.

[0144] 2-2. Current measurement based on glucose concentration

[0145] After adding a stirring rod to 50 mL of 10 mM PBS solution, the it curve was measured while stirring the electrode prepared in Example 2-1, which contained the redox polymer and glucose dehydrogenase, at 150 rpm. A certain amount of glucose dissolved in 10 mM PBS was added every 200 seconds, and the current was measured at concentrations of 0.01, 0.05, 0.1, 0.5, 1, 2, 4, 8, 16, 32, 64, and 100 mM to confirm the current changes. The results were... Figure 6a As shown in the image.

[0146] As a result of the tests, the current gradually increased as the glucose concentration in the electrodes gradually increased, with redox polymers 37 and 38 cross-linked with the enzyme, respectively. Firstly, in the current-time plot, both electrodes showed a gradual increase up to a concentration of 100 mM, and the change was significant even in the low concentration range (0–1 mM). Furthermore, when the current generated according to concentration was averaged and a current-concentration plot was plotted, the correlation with the magnitude of the current generated at each glucose concentration was confirmed. In the low concentration range, the constant current increased linearly, but as the concentration gradually increased, the current increase gradually decreased in the high concentration range of 16 mM or higher, indicating that the current value became constant. This shows that the maximum value at which glucose can be oxidized in the electrodes was reached, and the current increase gradually decreased, reaching the limiting catalytic current (i). max Furthermore, the electrode crosslinked with redox polymer 37 showed a maximum current of approximately 1.5 μA, while the electrode crosslinked with redox polymer 38 showed a maximum current of approximately 0.5 μA, which is one-third lower than that of the 37 type electrode.

[0147] After adding a stirring rod to 50 mL of 10 mM PBS solution, the it curve was measured while stirring the electrode prepared in Example 2-1, which contained the redox polymer and glucose dehydrogenase, at 150 rpm. In the electrode comprising redox polymer 1 and redox polymer 2, which included transition metal complexes containing CN ligands, enzyme-crosslinked solutions of redox polymer 1 and redox polymer 2 were added every 200 seconds. Current changes were measured at low concentrations (0.01, 0.05, 0.1, 0.5 mM) and high concentrations (1, 5, 10, 50, 100 mM) to confirm the current changes. Results are as follows: Figures 6b to 6c As shown in the image.

[0148] As a result of the tests, the current in both electrodes gradually increased in both the low concentration range (0–0.5 mM) and the high concentration range (0–100 mM). In the low concentration range, the constant current increased linearly, while in the high concentration range, the concentration increased gradually, showing that the current value became constant starting from a concentration of 50 mM or higher. This reached the maximum value at which glucose can be oxidized in the electrodes, and the electrode in which redox polymer 1 containing a transition metal complex with CN ligands was crosslinked showed a maximum current of about 1.0 μA, while the electrode in which redox polymer 2 containing a transition metal complex with CN ligands was crosslinked showed a maximum current of about 0.2 μA.

[0149] In addition, the current changes before and after cyclic voltammetry testing were measured. Specifically, the current changes were measured in 10 mM PBS at -0.5 to +0.1 V, and in the case of redox polymers 1 and 2 including electron transfer mediators containing CN ligands and PFPA, at -0.3 to +0.4 V, and the results were... Figures 7a to 7d As shown in [the image]. Figures 7a to 7d In this context, CV represents the value measured immediately after electrode completion, and the value after CV represents the value measured after stirring in PBS solution for 1 hour.

[0150] As described above, the redox polymer containing PFPA was well immobilized onto the enzyme and electrode surfaces, exhibiting a clear response even at low concentrations, and linear current changes were confirmed even at certain concentrations. Furthermore, the limiting catalytic currents of the cross-linked electrode were 1.5 μA, 0.5 μA, 1.0 μA, and 0.2 μA, respectively. This demonstrates that the immobilization method using TPMA-osmium complex-like electron transfer mediators or CN ligand-osmium complex-like electron transfer mediators and PFP polymer matrices can be used as a blood glucose sensor.

Claims

1. A redox polymer material for use in an electrochemical sensor, comprising a polypentafluorophenyl ester polymer for manufacturing the redox polymer material, having a structure of either chemical formula 1 or chemical formula 2: Chemical Formula 1 Chemical formula 2 In chemical formula 1 or chemical formula 2, R T and R L Each was independently selected from the group consisting of polydimethylacrylamide (PDMA) and polyacrylamide (PAA); and n is an integer from 10 to 300, and The transition metal complex is bonded to the polymer. The transition metal complex has the following structure, either chemical formula 3 or chemical formula 4: Chemical formula 3 Chemical Formula 4 In chemical formula 3 or chemical formula 4 M is a transition metal selected from the group consisting of: Os, Rh, Ru, Ir, Fe, and Co; and In chemical formula 3, L G1 and L G2 They combine with each other to form bidentate ligands selected from chemical formulas 5 to 6 below; and L G3 and L G4 They combine to form bidentate ligands selected from chemical formulas 5 to 6 below; and L G5 and L G6 They combine with each other to form bidentate ligands selected from chemical formulas 5 to 6 below; and Chemical Formula 5 Chemical Formula 6 In chemical formula 5, L C It is a heterocyclic compound comprising one or more nitrogen atoms, and is connected to the chemical formula of benzene at the 2-position; and In chemical formula 6, L N For heterocyclic compounds comprising one or more nitrogen atoms, and L N1 and L N2 Each of the two positions is connected to the other; and R L L is a heterocyclic compound C L N1 and L N2 All functional groups; and R1, R2, R3, R4, R5 and R L Each is independently selected from the group consisting of: substituted or unsubstituted alkyl groups having 1 to 10 carbon atoms, substituted or unsubstituted glycol groups having 2 to 20 carbon atoms, substituted or unsubstituted alcohol groups having 1 to 20 carbon atoms, substituted or unsubstituted haloalkyl groups having 1 to 20 carbon atoms, substituted or unsubstituted thiol groups having 1 to 20 carbon atoms, substituted or unsubstituted alkyl azide groups having 3 to 20 carbon atoms, substituted or unsubstituted aryl azide groups having 7 to 30 carbon atoms, substituted or unsubstituted alkenyl groups having 2 to 40 carbon atoms, substituted or unsubstituted alkynyl groups having 2 to 40 carbon atoms, cyano groups, haloyl groups, deuterium groups, and hydrogen groups; and L of chemical formula 4 H1 L H3 and L H4 With L H2 They bond together around a central point to form a tetradentate ligand represented by the following chemical formula 7; and Then, L H5 and L H6 They respectively form monodentate ligands of transition metal M represented by chemical formula 8; or L H5 and L H6 They combine with each other to form bidentate ligands of the transition metal M, as represented by the following chemical formula 9; and Chemical Formula 7 In chemical formula 7, C a C b and C c The compound is a heterocyclic compound comprising one or more nitrogen atoms, wherein an amino group and a methylene group are attached at the 2-position of the heterocyclic compound, and the three nitrogen atoms of the three heterocycles and the one nitrogen atom at the center of the three heterocycles connecting them are attached to a transition metal M, and R1, R2, and R3 are C. a C b C c All functional groups, and R1, R2 and R3 are each independently selected from the group consisting of: substituted or unsubstituted alkyl groups having 1 to 10 carbon atoms, substituted or unsubstituted glycol groups having 2 to 20 carbon atoms, substituted or unsubstituted alcohol groups having 1 to 20 carbon atoms, substituted or unsubstituted haloalkyl groups having 1 to 20 carbon atoms, substituted or unsubstituted thiol groups having 1 to 20 carbon atoms, substituted or unsubstituted alkyl azide groups having 3 to 20 carbon atoms, substituted or unsubstituted aryl azide groups having 7 to 30 carbon atoms, substituted or unsubstituted alkenyl groups having 2 to 40 carbon atoms, substituted or unsubstituted alkynyl groups having 2 to 40 carbon atoms, cyano groups, haloyl groups, deuterium groups and hydrogen groups; Chemical Formula 8 Chemical formula 9 In chemical formula 8, L H5 and L H6 Each is independently a monodentate ligand, and each is independently a heterocyclic compound comprising one or more -H, -F, -Cl, -Br, -I, -NO2, -NCCH3, -CO, -OH2, -NH3, or a nitrogen atom; and In chemical formula 9, L H5 -L H6 It is a bidentate ligand, and is catechol, acetylacetone, 2-pyridinecarboxylic acid, 2-pyridinecarboxamide, 2,2-bipyridine, or 2,2-bithiazole; and The L1 is independently selected from the group consisting of: substituted or unsubstituted alkylene groups having 1 to 20 carbon atoms, substituted or unsubstituted cycloalkylene groups having 1 to 20 carbon atoms, substituted or unsubstituted glycol groups having 2 to 30 carbon atoms, substituted or unsubstituted arylene groups having 6 to 30 carbon atoms, and substituted or unsubstituted heteroarylene groups having 3 to 30 carbon atoms; and The A a Select from the group consisting of amino and ammonium groups.

2. The redox polymer material according to claim 1, having a weight-average molecular weight in the range of 1,000 g / mol to 500,000 g / mol.

3. The redox polymer material according to claim 1, wherein the functional groups of the transition metal complex selected from the group consisting of primary amines, secondary amines, and ammonium groups are bonded to the polymer.

4. A method for preparing a redox polymer material thin film, comprising coating an electrode with the redox polymer material according to claim 1, and then curing the coated electrode.

5. A redox polymer material film prepared by the method according to claim 4.

6. An electrochemical biosensor comprising a redox polymer material film prepared by the method according to claim 4.

7. A sensing layer for an electrochemical biosensor, comprising: Enzymes capable of oxidizing and reducing liquid biological samples; and The redox polymer material for electrochemical sensors according to claim 1.

8. The sensing layer for an electrochemical biosensor according to claim 7, wherein the enzyme comprises One or more oxidoreductases selected from the group consisting of dehydrogenases, oxidases, and esterases; or One or more oxidoreductases selected from the group consisting of dehydrogenases, oxidases and esterases, and one or more cofactors selected from the group consisting of flavin adenine dinucleotide (FAD), nicotinamide adenine dinucleotide (NAD) and pyrroloquinoline quinone (PQQ).

9. An electrochemical biosensor, comprising the sensing layer for an electrochemical biosensor according to claim 8.

10. The electrochemical biosensor according to claim 9, wherein the sensor is a continuous glucose monitoring sensor.

11. An apparatus comprising the redox polymer material according to claim 1.

12. The device according to claim 11, wherein the device is an electrochemical biosensor.

13. The device of claim 12, wherein the device is an implantable device.

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