Pentafluorophenyl ester-containing polymers and electrochemical biosensors containing the same

Abstract

The present invention relates to a polypentafluorophenyl ester-based polymer used in the manufacture of electrochemical sensors, which has high reactivity with amine functional groups, is resistant to hydrolysis, and has generally good solubility in organic solvents. In addition, compared to polymers having similar structures, it has low problems with steric hindrance, toxicity, and side effects, and is particularly useful for insertable devices in which part of the sensor is inserted into the human body, such as continuous blood glucose monitoring sensors. [Drawing] Figure 1

Description

[Technical Field]

[0001] [Cross-reference to related applications] This application claims the benefit of priority based on Korean Patent Application No. 10-2020-0189140 filed on December 31, 2020, and all contents disclosed in the documents of this Korean patent application are incorporated herein by reference.

[0002] The present invention relates to a polymer containing a pentafluorophenyl ester that is useful for immobilizing an electron transfer mediator and an oxidoreductase on the surface of an electrode in a device such as an electrochemical biosensor, and an electrochemical biosensor containing the same. [Background technology]

[0003] Diabetes is a disease that occurs when high blood sugar levels persist for a long period of time, leading to complications such as cardiovascular disease, stroke, and kidney disease. In response to high blood sugar levels, it is necessary to inject insulin to lower blood sugar levels, but if too much insulin is injected, hypoglycemia can occur, which can lead to shock or death. To prevent this, diabetic patients must continuously monitor their blood sugar levels using a blood glucose sensor to maintain appropriate blood sugar levels.

[0004] Recently, blood glucose sensors that use the Continuous Glucose Monitoring System (CGMS) have been extensively researched and commercialized. CGMS sensors are devices that are inserted into the subcutaneous tissue and continuously measure glucose levels through interstitial fluid, not blood. While it is difficult to accurately ascertain changes in blood glucose levels with fingertip blood sampling, which requires measuring blood glucose five to six times a day, CGMS has the advantage of being able to check changes and trends in blood glucose levels throughout the day. This allows patients to quickly recognize hyperglycemia or hypoglycemia, further improving their blood glucose management capabilities.

[0005] A biosensor is a device that selectively detects biological samples and converts them into specific signals. Enzyme-based biosensors using electrochemical methods are particularly popular due to their improved selectivity, miniaturization, and measurement accuracy. Electrochemical blood glucose biosensors can be broadly divided into first-generation and second-generation types. First-generation blood glucose sensors, first developed by Clark and Lyon, measure blood glucose levels based on the decrease in oxygen concentration and the change in the concentration of hydrogen peroxide generated through an enzymatic oxidation-reduction reaction. Second-generation blood glucose sensors transfer electrons generated by the enzymatic oxidation-reduction reaction to the electrode via an electron transfer mediator. Second-generation sensors have many advantages over first-generation sensors, including less error due to oxygen concentration and a more efficient and rapid electron transfer reaction via the mediator. For these reasons, second-generation sensor types, including electron transfer mediators, are used in CGMS blood glucose sensors.

[0006] Electrochemical enzyme-based blood glucose biosensors generally consist of an enzyme, an electron transfer mediator, and an electrode. First, glucose is oxidized to gluconolactone by the enzyme, then the reduced enzyme is oxidized and donates electrons to the electron transfer mediator. Next, the reduced mediator is oxidized and transfers electrons to the electrode. This series of processes allows blood glucose levels to be confirmed through an electrical signal.

[0007] The method for immobilizing an electron transfer mediator and enzyme on the electrode surface is crucial for fabricating a blood glucose sensor. Previous studies have focused on immobilizing an enzyme on the electrode by mixing a redox hydrated polymer containing an electron transfer mediator with an enzyme and adding a crosslinker. Among these, polyvinylpyridine and polyvinylimidazole are well-known as redox polymer matrices. However, as shown in Reaction Scheme 1 below, these existing redox polymers have long and complicated synthesis steps for the final material, low immobilization efficiency of transition metal complexes, and difficulty in incorporating other functionalities into the polymer. Therefore, the development of new materials that overcome the limitations of existing materials is currently required to develop redox polymers with superior performance.

[0008] Under these circumstances, the present inventors have conducted extensive research into polymers useful for immobilizing electron transfer mediators and enzymes for electrochemical biosensors. As a result, they have found that when a polymer containing pentafluorophenyl (PFP) ester is used, it has high reactivity with amine functional groups, is resistant to hydrolysis, has generally good solubility in organic solvents, and has less steric hindrance and toxicity compared to polymers with similar structures. They have confirmed that this polymer is particularly useful for insertable devices in which part of the sensor is inserted into the human body, such as continuous glucose monitoring sensors, and have thus completed the present invention. Summary of the Invention [Problem to be solved by the invention]

[0009] An object of the present invention is to provide a polymer containing a pentafluorophenyl ester for use in preparing a redox polymeric material.

[0010] It is still another object of the present invention to provide a thin film of an oxidation-reduction polymer material and an electrochemical biosensor, which contain the transition metal complex and the polymer for preparing the oxidation-reduction polymer material. [Means for solving the problem]

[0011] In one aspect to achieve the above object, the present invention relates to a polymer for preparing an oxidation-reduction polymer material containing a pentafluorophenyl ester, an oxidation-reduction polymer for an electrochemical biosensor comprising the polymer and a transition metal complex, a oxidation-reduction polymer thin film containing the oxidation-reduction polymer for an electrochemical biosensor prepared from the polymer, and an electrochemical biosensor, e.g., a blood glucose sensor, including the oxidation-reduction polymer thin film. [Effects of the Invention]

[0012] When used in electrochemical sensors, the polypentafluorophenyl ester-based polymers of the present invention have high reactivity with amine functional groups, are resistant to hydrolysis, have high solubility in organic solvents, and are less susceptible to steric hindrance, toxicity, and side effects, making them particularly useful for insertable devices in which part of the sensor is inserted into the human body, such as continuous blood glucose monitoring sensors. [Brief explanation of the drawings]

[0013] [Figure 1] 1 is a graph showing the reaction monitoring results by 19F NMR after adding osmium complex 22 to PFPA polymer P1 according to the present invention. [Figure 2a] This is a graph showing the 19F NMR of the final reaction of the PFPA polymer (36) with the osmium complex (22, 33), and the predicted ratio of the redox polymer (37, 38) based on this. [Figure 2b] This is a graph showing the 19F NMR of the final reaction of the PFPA polymer (36) with the osmium complex (22, 33), and the predicted ratio of the redox polymer (37, 38) based on this. [Figure 3] 19F NMR data confirming the synthesis of a redox polymer including a transition metal complex containing a CN ligand and a PFP polymer in Examples 1-4. [Figure 4]19F NMR data confirming the synthesis of redox polymers including transition metal complexes containing CN ligands and PFP polymers in Examples 1-5. [Figure 5] Schematic diagram of the composition of the oxidation-reduction polymer and enzyme and the cross-linking method on the electrode surface of SPCEs. [Figure 6a] Graphs showing the change in current (top) when the glucose concentration was increased to 0.01, 0.05, 0.1, 0.5, 1, 2, 4, 8, 16, 32, 64, and 100 mM every 200 seconds, the change in current depending on the glucose concentration (bottom), the low concentration range (0 to 1 mM) (left), and the full range (0 to 100 mM) (right) for the oxidation-reduction polymers (37 and 38) of the present invention. [Figure 6b] 1 shows graphs illustrating the change in current (top left) and the change in current as a function of glucose concentration (top right) when the glucose concentration was increased to 0.01, 0.05, 0.1, and 0.5 mM every 200 seconds for an oxidation-reduction polymer according to the present invention (oxidation-reduction polymer 1 comprising a transition metal complex containing a CN ligand and a PFP polymer), as well as the change in current as a function of glucose concentration (bottom left) and the change in current as a function of glucose concentration (bottom right) when the glucose concentration was increased to 0, 1, 5, 10, 50, and 100 mM every 200 seconds. [Figure 6c] 1 shows graphs illustrating the change in current (top left) and the change in current as a function of glucose concentration (top right) when the glucose concentration is increased to 0.01, 0.05, 0.1, and 0.5 mM every 200 seconds for an oxidation-reduction polymer according to the present invention (oxidation-reduction polymer 1 comprising a transition metal complex containing a CN ligand and a PFP polymer), as well as the change in current as a function of glucose concentration (bottom left) and the change in current as a function of glucose concentration (bottom right) when the glucose concentration is increased to 0, 1, 5, 10, 50, and 100 mM every 200 seconds. [Figure 7a] 7A and 7B are graphs showing the results of measuring the change in current before and after cyclic voltammetry tests for redox polymers according to the present invention (redox polymers 1 and 2, which comprise transition metal complexes containing 37 and 38 and CN ligands, and a PFP polymer). The CV in Fig. 7A shows the value measured immediately after the electrode was assembled, while the "after" CV shows the value measured after stirring in PBS solution for 1 hour. [Figure 7b] 7b shows the results of measuring the change in current before and after cyclic voltammetry for redox polymers according to the present invention (redox polymers 1 and 2, which comprise transition metal complexes 37 and 38 and CN ligands, and PFP polymers). The CV in FIG. 7b shows the value measured immediately after electrode completion, and the "after CV" shows the value measured after stirring in PBS solution for 1 hour. [Figure 7c] 7c shows the results of measuring the change in current before and after cyclic voltammetry for redox polymers according to the present invention (redox polymers 1 and 2, which comprise transition metal complexes 37 and 38 and CN ligands, and PFP polymers). The CV in Fig. 7c shows the value measured immediately after electrode completion, while the "after" CV shows the value measured after stirring in PBS solution for 1 hour. [Figure 7d] 7(d) shows the results of measuring the change in current before and after cyclic voltammetry for redox polymers according to the present invention (redox polymers 1 and 2, which comprise transition metal complexes 37 and 38 and CN ligands, and PFP polymers). The CV in FIG. 7(d) shows the value measured immediately after the electrode was assembled, and the "after" CV shows the value measured after stirring in PBS solution for 1 hour. DETAILED DESCRIPTION OF THE INVENTION

[0014] The present invention will now be described in more detail.

[0015] The polymer for producing an oxidation-reduction polymer material according to the present invention is based on polypentafluorophenyl ester, specifically, it contains a repeating unit derived from pentafluorophenyl ester and is used as a polymer precursor for an oxidation-reduction polymer material. Preferably, the polymer can form an oxidation-reduction polymer material for an electron transfer mediator together with a crosslinker having a reactive group containing an amine group and a transition metal complex.

[0016] In one embodiment, the polymer for preparing an oxidation-reduction polymer according to the present invention may be a polymer for preparing an oxidation-reduction polymer, characterized in that a crosslinking agent containing an amine group is introduced into the polypentafluorophenyl ester by aminolysis using primary and secondary amine compounds.

[0017] Non-limiting examples of the polymer for the oxidation-reduction polymer material may be one or more polymers selected from the group consisting of polypentafluorophenyl acrylate (PPFPA) homopolymer, polypentafluorophenyl methacrylate (PPFPM) homopolymer, polypentafluorophenyl acrylate-polydimethylacrylamide (PPFPA-PDMA) copolymer, polypentafluorophenyl methacrylate-polydimethylacrylamide (PPFPM-PDMA) copolymer, polypentafluorophenyl acrylate-polyacrylamide (PPFPA-PAA) copolymer, and polypentafluorophenyl methacrylate-polyacrylamide (PPFPM-PAA) copolymer, but are not limited thereto.

[0018] In one embodiment, the polymer for preparing the redox polymer material according to the present invention may have the structure of the following formula 1 or 2: [Chemical formula 1] [ka] [Chemical formula 2] [ka] In the above Chemical Formula 1 or 2, R T and R Lare each independently selected from the group consisting of a substituted or unsubstituted alkylene group having 1 to 20 carbon atoms, a substituted or unsubstituted cycloalkylene group having 1 to 20 carbon atoms, a substituted or unsubstituted ethylene glycol group having 3 to 30 carbon atoms, a substituted or unsubstituted arylene group having 6 to 30 carbon atoms, a substituted or unsubstituted heteroarylene group having 3 to 30 carbon atoms, a substituted or unsubstituted alkenyl group having 2 to 40 carbon atoms, and a substituted or unsubstituted alkynyl group having 2 to 40 carbon atoms; The n is an integer of 10 to 300.

[0019] Unless otherwise specified in the present invention, "substituted" in the present invention means that at least one hydrogen atom is replaced with a halogen atom (e.g., F, Cl, Br, or I), a cyano group, a hydroxyl group, a thiol group, a nitro group, an amino group, an imino group, an azide group, an amidino group, a hydrazino group, a hydrazono group, an oxo group, a carbonyl group, a carbamyl group, an ester group, an ether group, a carboxyl group or a salt thereof, a sulfonic acid group or a salt thereof, a phosphoric acid group or a salt thereof, an alkyl group having 1 to 6 carbon atoms, a haloalkyl group having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, a haloalkenyl group having 2 to 6 carbon atoms, an alkoxy ... and aryl groups having 1 to 9 carbon ring members, heterocycloalkyl groups having 1 to 9 carbon ring members, aryl groups having 6 to 10 carbon atoms, aryloxy groups having 6 to 10 carbon atoms, arylthio groups having 6 to 10 carbon atoms, heteroalkyl groups having 1 to 9 carbon ring members, heteroaryloxy groups having 1 to 9 carbon ring members, and heteroarylthio groups having 1 to 9 carbon ring members.

[0020] Preferably, the R T and R Lmay each independently be selected from the group consisting of polydimethylacrylamide (PDMA), polyacrylamide (PAA), polystyrene (PS), polyethylene glycol (PEG), polyethylene oxide (PEO), polymethyl methacrylate (PMMA), polyvinylimidazole (PVI), polyvinylpyridine (PVP), and polysiloxane (PDMS), each having a molecular weight of 1,000 g / mol to 50,000 g / mol.

[0021] For example, the polymer for preparing the oxidation-reduction polymer material according to the present invention may have a weight average molecular weight ranging from 1,000 g / mol to 500,000 g / mol, but is not limited thereto.

[0022] In another aspect, the present invention relates to an oxidation-reduction polymer material for an electrochemical sensor, in which a transition metal complex is introduced into the polypentafluorophenyl ester-based polymer.

[0023] For example, such an oxidation-reduction polymer material can be produced by functionalizing the polypentafluorophenyl ester polymer by introducing a compound containing a functional group selected from the group consisting of an amine group, an ammonium group, a halogen group, an epoxy group, an azide group, an acrylate group, an alkenyl group, an alkynyl group, a thiol group, an isocyanate group, an alcohol group, and a silane group, and then bonding a transition metal complex to the functionalized polymer.

[0024] Alternatively, the oxidation-reduction polymer material may be prepared by functionalizing the polypentafluorophenyl ester-based polymer by introducing functional groups such as the crosslinking substances, such as amine groups, ammonium groups, thiol groups, and alcohol groups, into the ligands of a transition metal complex through substitution and addition reactions, and then bonding the functionalized transition metal complex with the polymer according to the present invention.

[0025] Alternatively, the polymer material can be prepared by functionalizing and bonding the polypentafluorophenyl ester-based polymer and the transition metal complex together using a crosslinking material containing a functional group selected from the group consisting of an amine group, an ammonium group, a halogen group, an epoxy group, an azide group, an acrylate group, an alkenyl group, an alkynyl group, a thiol group, an isocyanate group, an alcohol group, and a silane group.

[0026] Preferably, the oxidation-reduction polymer material according to the present invention may be a polypentafluorophenyl ester-based polymer to which a functional group selected from the group consisting of a primary and secondary amine group and an ammonium group and a transition metal complex are bound.

[0027] Specifically, the transition metal complex may have a structure represented by the following formula 3 or 4: [Chemical formula 3] [ka] [Chemical formula 4] [ka] In the above Chemical Formula 3 or 4, M is a transition metal selected from the group consisting of Os, Rh, Ru, Ir, Fe, and Co; In the above Chemical Formula 3, L G1 and L G2 are combined with each other to form a bidentate ligand selected from the following chemical formulas 5 and 6: L G3 and L G4 are combined with each other to form a bidentate ligand selected from the following chemical formulas 5 and 6; L G5 and L G6 are combined with each other to form a bidentate ligand selected from the following chemical formulas 5 and 6: [Chemical formula 5] [ka] [Chemical formula 6] [ka] In the above Chemical Formula 5, L C is a heterocyclic compound containing one or more nitrogen atoms, and can be linked to the benzene formula, preferably at the 2-position; In the above Chemical Formula 6, L N is a heterocyclic compound containing one or more nitrogen atoms, and preferably L N1 and L N2 are linked to each other at their second positions.

[0028] R L is a heterocyclic compound L C , L N1 and L N2 are all functional groups.

[0029] In one embodiment, R1, R2, R3, R4, R5 and R L are each independently selected from the group consisting of a substituted or unsubstituted alkyl group having 1 to 10 carbon atoms, a substituted or unsubstituted ethylene glycol group having 2 to 20 carbon atoms, a substituted or unsubstituted alcohol group having 1 to 20 carbon atoms, a substituted or unsubstituted alkyl halogen group having 1 to 20 carbon atoms, a substituted or unsubstituted thiol group having 1 to 20 carbon atoms, a substituted or unsubstituted alkyl azide group having 3 to 20 carbon atoms, a substituted or unsubstituted aryl azide group having 7 to 30 carbon atoms, a substituted or unsubstituted alkenyl group having 2 to 40 carbon atoms, a substituted or unsubstituted alkynyl group having 2 to 40 carbon atoms, a cyano group, a halogen group, deuterium, and hydrogen.

[0030] In one embodiment, L of Formula 4 H1 , L H3 and L H4 L H2 can combine with each other at the center to form a tetradentate ligand represented by the following chemical formula 7.

[0031] At this time, L H5 and L H6 form monodentate ligands for the transition metal M as shown in formula 8; L H5 and L H6are combined with each other to form a bidentate ligand for the transition metal M as represented by the following chemical formula 9: [Chemical formula 7] [ka] In the above Chemical Formula 7, C a , C b and C c are each independently a heterocyclic compound containing one or more nitrogen atoms, and preferably, the heterocyclic compound can be linked to methylamine at its 2-position, and three nitrogen atoms in the three heterocyclic rings and one central nitrogen atom linking the three heterocyclic rings together can be linked to a transition metal M.

[0032] R1, R2, and R3 are heterocyclic compounds C a , C b , C c means all functional groups of

[0033] In one embodiment, R1, R2, and R3 are each independently selected from the group consisting of a substituted or unsubstituted alkyl group having 1 to 10 carbon atoms, a substituted or unsubstituted ethylene glycol group having 2 to 20 carbon atoms, a substituted or unsubstituted alcohol group having 1 to 20 carbon atoms, a substituted or unsubstituted alkylhalogen group having 1 to 20 carbon atoms, a substituted or unsubstituted thiol group having 1 to 20 carbon atoms, a substituted or unsubstituted alkylazide group having 3 to 20 carbon atoms, a substituted or unsubstituted arylazide group having 7 to 30 carbon atoms, a substituted or unsubstituted alkenyl group having 2 to 40 carbon atoms, a substituted or unsubstituted alkynyl group having 2 to 40 carbon atoms, a cyano group, a halogen group, deuterium, and hydrogen. Preferably, a reactive group capable of linking to a polymer can be introduced into each of them independently. [Chemical formula 8] [ka] [Chemical formula 9] [ka]

[0034] In the above Chemical Formula 8, L H5 and L H6 are each monodentate ligands and may each independently be -H, -F, -Cl, -Br, -I, -NO2, -NCCH3, -CO, -OH2, -NH3, or a heterocyclic compound containing one or more nitrogen atoms.

[0035] In the above Chemical Formula 9, L H5 -L H6 is a bidentate ligand and can be catechol, acetylacetone, 2-picolinic acid, 2-pyridinecarboxamide, 2,2-bipyridine, or 2,2-bithiazole.

[0036] L1 is independently selected from the group consisting of a substituted or unsubstituted alkylene group having 1 to 20 carbon atoms, a substituted or unsubstituted cycloalkylene group having 1 to 20 carbon atoms, a substituted or unsubstituted ethylene glycol group having 2 to 30 carbon atoms, a substituted or unsubstituted arylene group having 6 to 30 carbon atoms, and a substituted or unsubstituted heteroarylene group having 3 to 30 carbon atoms; The above A a is selected from the group consisting of an amine group and an ammonium group.

[0037] One aspect of the present invention relates to a method for manufacturing an oxidation-reduction polymer thin film, which includes coating an electrode with an oxidation-reduction polymer material containing the polymer and then curing the coated electrode, as well as an electrochemical biosensor including the oxidation-reduction polymer thin film manufactured thereby.

[0038] Another aspect of the present invention relates to a sensing film for an electrochemical biosensor, which comprises an enzyme capable of oxidizing and reducing a liquid biological sample and an electron transfer mediator including the transition metal complex.

[0039] Oxidoreductase is a general term for enzymes that catalyze oxidation-reduction reactions in living organisms. In the present invention, it refers to an enzyme that reacts with and reduces a target substance to be measured, such as a target substance in the case of a biosensor. The reduced enzyme reacts with an electron transfer mediator, and the target substance is quantified by measuring a signal, such as a change in current, generated during this reaction. The oxidoreductase that can be used in the present invention can be one or more enzymes selected from the group consisting of various dehydrogenases, oxidases, esterases, etc. Depending on the oxidation-reduction or target substance to be detected, an enzyme that uses the target substance as a substrate can be selected from the above enzyme group and used.

[0040] More specifically, the oxidoreductase 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, bilirubin oxidase, and the like.

[0041] Meanwhile, the oxidoreductase may contain a cofactor that serves to store the hydrogen removed by the oxidoreductase from the target substance (e.g., target substance) to be measured. For example, the cofactor may be one or more selected from the group consisting of flavin adenine dinucleotide (FAD), nicotinamide adenine dinucleotide (NAD), pyrroloquinoline quinone (PQQ), etc.

[0042] For example, when measuring the glucose concentration in blood, glucose dehydrogenase (GDH) can be used as the oxidoreductase, and the glucose dehydrogenase can be flavin adenine dinucleotide-glucose dehydrogenase (FAD-GDH) containing FAD as a cofactor and / or nicotinamide adenine dinucleotide-glucose dehydrogenase containing FAD-GDH as a cofactor.

[0043] Specific examples of the usable oxidoreductase include 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.), ascorbate 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.), bilirubin oxidase (e.g., EC 1.3.3.5, etc.) and the like.

[0044] Most preferably, the oxidoreductase is a glucose dehydrogenase that can maintain 70% or more of its activity in a buffer solution at 37°C for one week.

[0045] The sensing membrane 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 the oxidation-reduction polymer based on 100 parts by weight of the oxidation-reductase. The content of the oxidation-reduction polymer can be appropriately adjusted depending on the activity of the oxidation-reductase.

[0046] Furthermore, the sensing film according to the present invention may further include carbon nanotubes to enhance the film performance. Specifically, when carbon nanotubes are used together with a transition metal complex, especially osmium, the electron transfer rate increases, thereby further enhancing the performance of the sensing film.

[0047] The sensing membrane according to the present invention may further include a cross-linking agent.

[0048] Meanwhile, the sensing membrane according to the present invention may further contain one or more additives selected from the group consisting of surfactants, water-soluble polymers, quaternary ammonium salts, fatty acids, thickeners, etc., to serve as a dispersant when dissolving the reagent, an adhesive when preparing the reagent, a stabilizer for long-term storage, etc.

[0049] The surfactant may help the composition to spread evenly and uniformly on the electrode when dispensed. The surfactant may be at least one selected from the group consisting of Triton X-100, sodium dodecyl sulfate, perfluorooctane sulfonate, sodium stearate, etc. The reagent composition according to the present invention may contain the surfactant in an amount of 3 to 25 parts by weight, e.g., 10 to 25 parts by weight, based on 100 parts by weight of the oxidoreductase, to ensure that the reagent spreads evenly and uniformly on the electrode when dispensed. For example, when an oxidoreductase with an activity of 700 U / mg is used, the surfactant may be added in an amount of 10 to 25 parts by weight based on 100 parts by weight of the oxidoreductase. If the activity of the oxidoreductase is higher than this, the surfactant content may be adjusted to a lower amount.

[0050] The water-soluble polymer may serve as a polymer support for the reagent composition, helping to stabilize and disperse the enzyme. The water-soluble polymer may be at least one selected from the group consisting of polyvinyl pyrrolidone (PVP), polyvinyl alcohol (PVA), polyperfluorosulfonate, hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), carboxymethyl cellulose (CMC), cellulose acetate, and polyamide. The reagent composition according to the present invention may contain the water-soluble polymer in an amount of 10 to 70 parts by weight, for example, 30 to 70 parts by weight, based on 100 parts by weight of the redox enzyme, to adequately and appropriately stabilize and disperse the redox enzyme. For example, when an oxidoreductase with an activity of 700 U / mg is used, the water-soluble polymer may be contained in an amount of 30 to 70 parts by weight based on 100 parts by weight of the oxidoreductase. If the activity of the oxidoreductase is higher than this, the content of the water-soluble polymer may be adjusted to be lower than this.

[0051] The water-soluble polymer may have a weight-average molecular weight of about 2,500 g / mol to 3,000,000 g / mol, for example, about 5,000 g / mol to 1,000,000 g / mol, in order to effectively stabilize and disperse the support and enzyme.

[0052] The thickener serves to firmly attach the reagent to the electrode. Examples of the thickener include at least one selected from the group consisting of Natrosol, diethylaminoethyl-dextran hydrochloride (DEAE-Dextran hydrochloride), and the like. The electrochemical sensor according to the present invention may contain the thickener in an amount of 10 to 90 parts by weight, e.g., 30 to 90 parts by weight, based on 100 parts by weight of the oxidoreductase, to firmly attach the oxidation-reduction polymer according to the present invention to the electrode. For example, when an oxidoreductase having an activity of 700 U / mg is used, the sensor may contain 30 to 90 parts by weight of the thickener based on 100 parts by weight of the oxidoreductase. If the activity of the oxidoreductase is higher than this, the amount of the thickener may be adjusted to be lower.

[0053] In another aspect, the present invention provides a device, preferably an insertable device, such as a device insertable into the human body, that includes such an organic electron transfer mediator. The device may also preferably be an electrochemical biosensor, more preferably an electrochemical glucose (blood sugar) sensor.

[0054] Specifically, there is no limitation on the type of the electrochemical biosensor, but it may preferably be a continuous blood glucose monitoring sensor.

[0055] The continuous glucose monitoring sensor of the present invention may include, for example, an electrode, an insulator, a substrate, a sensing layer containing the oxidation-reduction polymer and the redox enzyme, a diffusion layer, and a protection layer. The electrodes may include two types of electrodes, such as a working electrode and a counter electrode, or three types of electrodes, such as a working electrode, a counter electrode, and a reference electrode. In one embodiment, the biosensor of the present invention may be an electrochemical biosensor fabricated by coating a substrate having at least two, preferably two or three, electrodes with a reagent composition containing the oxidation-reduction polymer containing the organic electron transfer mediator of Formula 1 and an enzyme capable of oxidizing and reducing a liquid biological sample, followed by drying. For example, a planar electrochemical biosensor is provided, in which a working electrode and a counter electrode are provided on opposite sides of a substrate, a sensing layer containing the oxidation-reduction polymer having the organic electron transfer mediator of the present invention is laminated on the working electrode, and an insulator, a diffusion layer, and a protection layer are laminated, in order, on both sides of the substrate on which the working electrode and counter electrode are provided.

[0056] In a specific embodiment, the substrate may be made of one or more materials selected from the group consisting of PET (polyethylene terephthalate), PC (polycarbonate), and PI (polyimide).

[0057] The working electrode may be a carbon, gold, platinum, silver, or silver / silver chloride electrode.

[0058] In addition, in the case of an electrochemical biosensor having two electrodes, the counter electrode also serves as the reference electrode, so a gold, platinum, silver, or silver / silver chloride electrode can be used as the counter electrode, and in the case of an electrochemical biosensor having three electrodes including a reference electrode, a gold, platinum, silver, or silver / silver chloride electrode can be used as the reference electrode, and a carbon electrode can be used as the counter electrode.

[0059] The diffusion membrane may be made of Nafion, cellulose acetate, or silicone rubber, and the protective membrane may be made of silicone rubber, polyurethane, or polyurethane copolymer, but is not limited thereto.

[0060] As a non-limiting example, in the case of a two-electrode system, the counter electrode also serves as the reference electrode, so silver chloride or silver can be used, and in the case of a three-electrode system, the reference electrode can be silver chloride or silver, and the counter electrode can be a carbon electrode.

[0061] Although the specific example of the present invention illustrates a biosensor for measuring glucose as an example of an applicable electrochemical biosensor, by varying the type of enzyme contained in the reagent composition of the present invention, the present invention can be applied to a biosensor for quantifying various substances such as cholesterol, lactate, creatinine, hydrogen peroxide, alcohol, amino acids, and glutamate. [Example]

[0062] The present invention will be described in more detail below with reference to the following examples, but the following examples are merely illustrative of the present invention and are not intended to limit the scope of the present invention.

[0063] Experimental materials Reference Example 1. Reagents and samples

[0064] Commercially purchased reagents from Aldrich, Acros, TCI, and Alfa Aesar were used without further purification. Ethylenediamine, pyridine, 1-methylimidazole, and triethylamine were used after filtering a 10 mL pipette filled with basic alumina. The following solvents, purchased from Daejung and Samjun, were used after purification: anhydrous dichloromethane and anhydrous N,N-dimethylformamide were obtained by distillation over calcium hydride, and anhydrous acetonitrile was obtained by distillation over P2O5. Diethyl ether was distilled over sodium and benzophenone. Analytical thin-layer chromatography was performed using Merck silica gel 60 F. 254 Fluorescence was observed using a glass plate with a dual short-wavelength (254 nm) / long-wavelength (365 nm) UV lamp. Column chromatography was performed using Aldrich basic aluminum oxide or Alfa neutral aluminum oxide. Zensor SE102 screen-printed carbon electrodes (SPCEs) were used.

[0065] Reference example 2. Equipment

[0066] Hydrogen nuclear magnetic resonance ( 1 H NMR), carbon nuclear magnetic resonance ( 13 C NMR) and fluorine nuclear magnetic resonance ( 19 F NMR spectra were obtained using a Varian Inova 400 (400 MHz for 1 H, and 100 MHz for 13 All chemical transfers were measured using a δ 0.00 spectrometer and tetramethylsilane (δ 7.26 for CDCl3 in 1 H NMR, δ77.2 for CDCl3in 13 C NMR), deuterated dimethyl sulfoxide (δ 2.50 for DMSO in 1 H NMR, δ39.52 for DMSO in 13C NMR), deuterated acetonitrile (δ 1.94 for CD3CN in 1 The values are shown in ppm based on 1H NMR. 19 In the case of F NMR, deuterated methanol is converted to trifluorotoluene (CF3-Ar, δ-69.27 for CD3OD in 19 The values are expressed in ppm using a small amount of F NMR as the reference. Cyclic voltammograms (CVs) were measured using a CH Instruments CHI1040C model. A well-cleaned 3 mm diameter carbon glass electrode was used as the working electrode, an Ag / AgCl electrode as the reference electrode, and a Pt electrode as the counter electrode, with scan rates of 10–50 mV / s. Mass spectra were obtained at the Sogang University Organic Chemistry Research Center using a ThermoFisher Scientific LTQ XL model ESI-Iontrap mass analyzer for low resolution and an ESI-orbitrap mass analyzer for high resolution. Single-crystal X-ray diffraction analysis was performed at the Korea Research Institute of Chemical Technology using a Bruker SMART APEX II model to determine the crystal structure.

[0067] Example 1. Synthesis of redox polymers using PFPA polymer matrix The purpose of this experiment was to synthesize a redox polymer by reacting a pentafluorophenyl acrylate (PFPA)-based polymer matrix with an electron transfer mediator containing an amine (-NH2) group at the terminal. First, a DMA:PFPA polymer (P1) was synthesized by radical polymerization of pentafluorophenyl acrylate (PFPA) and dimethylacrylamide (DMA) in a 5:5 ratio using AIBN.

[0068] 1-1.Synthesis of Poly(N,N-dimethyl acrylamide-co-pentafluorophenyl acrylate) (P1) [ka]

[0069] [Synthesis of DMA:PFPA polymer] N,N-dimethylacrylamide (150 mg, 1.51 mmol), pentafluorophenyl acrylate (360 mg, 1.51 mmol), and azobisisobutyronitrile (AIBN, 7 mg, 0.043 mmol) were placed in a glass culture tube, and acetonitrile (8 mL) was added to prepare a mixed solution. Argon gas was then bubbled through the tube for 10 minutes to create an argon atmosphere, and the mixture was then refluxed at 80°C for 12 hours. After the reaction was complete, the solvent was removed using a rotary evaporator, and CHCl was added to prepare a mixed solution. The resulting polymer was purified by slowly dropping the mixed solution into n-hexane to obtain a precipitate, which was repeated three times. The precipitate was filtered and washed with n-hexane, and the resulting polymer was dried under vacuum for at least 6 hours to obtain product P1 as a white solid. (482 mg, 95%) 1 H NMR (400 MHz, CDCl3) δ 2.81 to 3.34 ppm (br, 7H, -CH- in the backbone of PPFPA and -N(CH3)2from PDMA), 2.47 to 2.65 ppm (br, 1H, -CH- in the backbone of PDMA), 1.13 to 2.22 ppm (br, 4H, -CH2- in the backbone of PDMA and PPFPA), 19 F NMR (400 MHz, CDCl3) δ -152.90 (br, ortho), -157.21 (br, para), -162.15 (br, meta)

[0070] The synthesized polymer showed good solubility in methanol, acetonitrile, acetone, and dichloromethane, but was completely insoluble in water and diethyl ether.

[0071] 1-2. Confirmation of whether PFPA polymer can function as a polymer matrix capable of immobilizing electron transfer mediators and GDH In order to confirm whether or not the reaction between the polymer P1 synthesized in Example 1-1 and the osmium complex 22 containing an amine group at its terminal proceeded, 19F NMR was used to observe the structure. [ka]

[0072] Complex 22 has a zwitterion of Cl - The NMR peak was confirmed in deuterated methanol, which can dissolve the complex and polymer in the form of a deuterated methanol solvent. In addition, the amine group of complex 22 is ammonium (-NH4 + Since the complex cannot react with the PFP ester when it exists in the form of tetraethylamine (TEA), an equivalent amount of triethylamine (TEA) was added to the complex. The results are shown in Figure 1. As can be seen in Figure 1, first 19 F NMR analysis of polymer P1 alone revealed three broad peaks corresponding to PFPA. Next, upon addition of complex 22, three additional peaks of sharply pointed pentafluorophenol (PFPOH) were observed. These are PFPOH peaks, a by-product formed by the reaction of the amine at the end of complex 22. Over time, the PFPOH peaks increased in size, while the PFPA peaks decreased. After approximately 36 hours, all PFPA peaks disappeared, leaving only the PFPOH peaks. This suggests that PFP esters can react with the amine groups contained in the TPMA-based osmium complex, and that the PFPA polymer can function as a polymer matrix capable of immobilizing the electron transfer mediator and GDH.

[0073] 1-3. Synthesis of Redox Polymers polymers (36) and (37) Oxidation-reduction polymers were synthesized by reacting TPMA-based osmium complexes 22 and 33, which contain amine functional groups, with PFPA polymer 36 (Figure 2). The reaction was carried out by calculating the equivalent ratio to obtain an oxidation-reduction polymer with an osmium complex content of approximately 25–30% in the entire polymer chain.

[0074] 1) Synthesis of oxidation-reduction polymer (36) [ka]

[0075] In an NMR tube, starting material 22 (6 mg, 0.009 mmol) was dissolved in deuterated methanol (0.5 mL). The solution was added to starting material P1 (5 mg) dissolved in deuterated methanol (0.5 mL), followed by triethylamine (1.24 μL, 0.009 mmol). The reaction progress was monitored. 19 After confirming the reaction by F NMR, the polymer was purified by slowly dropping the reaction solution into diethyl ether three times to obtain a precipitate. The precipitate was filtered, washed with diethyl ether, and then dried under vacuum for more than 4 hours to obtain product 36 as a brown solid (6 mg, 64%). 19 F NMR (400 MHz, CD3OD) δ -153.38 (br, ortho), -159.74 (br, para), -163.53 (br, meta)

[0076] 2) Synthesis of oxidation-reduction polymer (37) [ka]

[0077] In an NMR tube, starting material 33 (6.5 mg, 0.009 mmol) was dissolved in deuterated methanol (0.5 mL). The solution was added to starting material P1 (5 mg) dissolved in deuterated methanol (0.5 mL), followed by triethylamine (1.22 μL, 0.009 mmol). The reaction progress was monitored. 19 After confirming the reaction by F NMR, the polymer was purified by slowly dropping the reaction solution into diethyl ether three times to obtain a precipitate. The precipitate was filtered, washed with diethyl ether, and dried under vacuum for more than 4 hours to obtain product 37 as a brown solid (7.2 mg, 73%). 19 F NMR (400 MHz, CD3OD) δ -154.57 (br, ortho), -160.27 (br, para), -164.95 (br, meta)

[0078] The reaction between the added complex and the PFPA polymer is 19 The ratio of the redox polymer was determined by F NMR until no further changes in the peaks occurred. It took approximately 24 to 36 hours for all the added osmium complex to react. 19 The ratios of the two redox polymers were confirmed by F NMR. Redox polymer 37 had a 2:3 PFPA:PFPOH ratio, with the osmium complex accounting for 30% of the total chain. Redox polymer 38 had a 2.2:2.8 PFPA:PFPOH ratio, indirectly indicating that approximately 28% of the osmium complex was contained in the polymer matrix. Both polymers were obtained by precipitation in diethyl ether. Unlike the starting polymer 36, both redox polymers were soluble in water. Furthermore, as confirmed below, CV measurements showed similar redox potentials to the osmium complex monomer. These results indicate that we have successfully synthesized redox polymers containing TPMA-based osmium complexes.

[0079] 1-4. Synthesis of transition metal complexes containing CN ligands and redox polymers containing PFP polymers 1 [ka]

[0080] In an NMR tube, the starting material, an osmium complex containing a CN ligand, [Os(2-(2-pyridinyl-κN)-5-methaneamine-phenyl-κC)(4,4'-dimethyl-2,2'-bipyridine)2]Cl2 (11 mg, 0.014 mmol), was dissolved in deuterated methanol (0.5 mL). The solution was added to the starting material, PFP polymer (7:3) (17 mg) dissolved in deuterated methanol (0.5 mL), followed by triethylamine (3.8 μL, 0.028 mmol). The progress of the reaction was monitored. 19After confirming the results by F NMR (Figure 3), the polymer was purified by slowly dropping the reaction solution into diethyl ether three times to obtain a precipitate. The precipitate was filtered, washed with diethyl ether, and dried under vacuum for more than 4 hours to obtain a brown solid product.

[0081] 1-5. Synthesis of redox polymers containing transition metal complexes with CN ligands 2 [ka]

[0082] In an NMR tube, dissolve the starting material [Os(2-(2-pyridinyl-κN)-5-methaneamine-phenyl-κC)(4,4'-dimethoxy-2,2'-bipyridine)2]Cl2 (9.7 mg, 0.012 mmol) in deuterated methanol (0.5 mL). Add the dissolved solution to the starting material PFP polymer (7:3) (15 mg) dissolved in deuterated methanol (0.5 mL), followed by triethylamine (3.1 μL, 0.024 mmol). Check the reaction progress. 19 After confirming the reaction by F NMR (Figure 4), the polymer was purified by slowly dropping the reaction solution into diethyl ether three times to obtain a precipitate. The precipitate was filtered, washed with diethyl ether, and dried under vacuum for more than 4 hours to obtain a brown solid product.

[0083] Example 2. Immobilization of an oxidation-reduction polymer and an enzyme and a glucose sensitivity test The following test was carried out to confirm the sensitivity to glucose concentration by immobilizing a solution of a mixture of an osmium complex-containing redox polymer and glucose dehydrogenase (GDH) on an SPCEs electrode.

[0084] 2-1. Fabrication of an electrode with an immobilized redox polymer and sugar dehydrogenase A mixed solution was prepared by dissolving 0.4 mg of redox polymer in 50 μL of distilled water and 0.8 mg of glucose dehydrogenase (GDH) in 50 μL of distilled water (v / v 1:1). This mixed solution was uniformly applied to SPCEs (0.83 μL for redox polymers 37 and 38, and 1 μL for redox polymers 1 and 2 containing transition metal complexes with CN ligands) and slowly dried at room temperature. After drying was complete, 0.83 μL of the mixed solution was applied to redox polymers 37 and 38, and 1 μL for redox polymers 1 and 2 containing transition metal complexes with CN ligands. This process was repeated twice more. A total of 2.5 μL (polymers 37 and 38) and 3 μL (redox polymers 1 and 2 containing transition metal complexes with CN ligands) of the mixed solution were applied to the electrodes, and the electrodes were dried at room temperature for at least 12 hours. The composition of the mediator and enzyme and the method of cross-linking on the SPCEs electrode surface are shown schematically in Figure 5.

[0085] 2-2. Measurement of current due to glucose concentration The electrode with the redox polymer and glucose dehydrogenase immobilized as prepared in Example 2-1 was placed in 50 mL of 10 mM PBS solution with a stir bar and stirred at 150 rpm to measure the I / O curve. A fixed 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 change in current. The results are shown in Figure 6a.

[0086] The test results showed that the electrodes with the 37 and 38 oxidation-reduction polymers cross-linked to the enzyme showed a gradual increase in current as the glucose concentration increased. First, the current-time graph showed that both electrodes showed a gradual, stepwise increase up to a concentration of 100 mM, and this change was also clearly evident in the low concentration range (0-1 mM). Furthermore, when the average current generated according to concentration was plotted as a current-concentration graph, a correlation was confirmed between the magnitude of the current generated by each glucose concentration. In the low concentration range, the amount of 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, and the current value remained constant. This is because the electrode reached the maximum value at which sugar could be oxidized, and the increase in current gradually decreased, resulting in a limiting catalytic current (i max ) was reached. Furthermore, the electrode cross-linked with the 37th oxidation-reduction polymer showed a maximum current of about 1.5 μA, while the electrode cross-linked with the 38th oxidation-reduction polymer showed a maximum current of about 0.5 μA, which was one-third lower than the 37th electrode.

[0087] The electrode with the oxidation-reduction polymer and glucose dehydrogenase immobilized prepared in Example 2-1 was placed in 50 mL of 10 mM PBS solution with a stir bar and stirred at 150 rpm, and an I-T curve was measured. For the electrode with the enzyme crosslinked to oxidation-reduction polymers 1 and 2 containing transition metal complexes with CN ligands, a fixed amount of glucose dissolved in 10 mM PBS was added every 200 seconds. The current was measured under low concentration ranges of 0.01, 0.05, 0.1, and 0.5 mM, and under high concentration ranges of 1, 5, 10, 50, and 100 mM, respectively, to confirm the change in current. The results are shown in Figures 6b and 6c.

[0088] The test results showed that both electrodes showed a gradual, stepwise increase in current magnitude in both the low concentration range (0-0.5 mM) and the high concentration range (0-100 mM). In the low concentration range, the current increased linearly, maintaining a constant amount, whereas in the high concentration range, the concentration gradually increased, maintaining a constant current value above 50 mM. This indicates that the electrode reached the maximum value at which sugar can be oxidized. The electrode crosslinked with oxidation-reduction polymer 1 containing a transition metal complex with a CN ligand showed a maximum current of approximately 1.0 μA, while the electrode crosslinked with oxidation-reduction polymer 2 containing a transition metal complex with a CN ligand showed a maximum current of approximately 0.2 μA.

[0089] Furthermore, we measured the current change before and after the cyclic voltage-current test. Specifically, the current change was measured from -0.5 to +0.1 V in 10 mM PBS, and from -0.3 to +0.4 V for the electron transfer mediator containing the CN ligand and the redox polymers 1 and 2 containing PFPA. The results are shown in Figures 7a to 7d. In Figures 7a to 7d, the CVs were measured immediately after the electrode was assembled, while the "after" CVs were measured after stirring in PBS for 1 hour.

[0090] As shown above, the redox polymer containing PFPA was well immobilized on the enzyme and electrode surface, demonstrating clear sensitivity even at low concentrations and a linear current change up to a certain concentration. Furthermore, the cross-linked electrode showed limiting catalytic currents of 1.5 μA, 0.5 μA, 1.0 μA, and 0.2 μA at a concentration of 100 mM. This demonstrates that the immobilization method using a TPMA-osmium complex-based electron transfer mediator or a CN ligand-osmium complex-based electron transfer mediator with a PFPA polymer matrix can function as a blood glucose sensor.

Claims

1. An oxidation-reduction polymer material for an electrochemical sensor, comprising a polypentafluorophenyl ester-based polymer having the structure of the following chemical formula 1 or 2: the electrochemical sensor is a continuous glucose monitoring device; The polymer is characterized in that a functional group selected from the group consisting of a primary and secondary amine group and an ammonium group and a transition metal complex are bound to the polymer, and the transition metal complex has the structure of the following chemical formula 3 or 4: [Chemical formula 1] 【Chemical 1】 [Chemical formula 2] 【Chemistry 2】 In the above Chemical Formula 1 or 2, R Ｔ and R Ｌ are each independently selected from the group consisting of polydimethylacrylamide (PDMA) and polyacrylamide (PAA); [Chemical formula 3] 【Chemistry 3】 [Chemical formula 4] 【Chemistry 4】 In the above Chemical Formula 3 or 4, M is a transition metal selected from the group consisting of Os, Rh, Ru, Ir, Fe, and Co; In the formula 3, L G1 and L G2 combine with each other to form a bidentate ligand selected from the following formulas 5 and 6: L G3 and L G4 combine with each other to form a bidentate ligand selected from the following chemical formulas 5 and 6: L G5 and L G6 combine with each other to form a bidentate ligand selected from the following chemical formulas 5 and 6; [Chemical formula 5] 【Chemistry 5】 [Chemical formula 6] 【Chemistry 6】 In Formula 5, L C is a heterocyclic compound containing one or more nitrogen atoms, and is connected to the benzene ring at the 2-position; In Formula 6, L N is a heterocyclic compound containing one or more nitrogen atoms, and L N1 and L N2 are connected to each other at the 2-position; R L is any functional group of the heterocyclic compounds L C , L N1 and L N2 ; wherein R 1 , R 2 , R 3 , R 4 , R 5 and R L are each independently selected from the group consisting of a substituted or unsubstituted alkyl group having 1 to 10 carbon atoms, a substituted or unsubstituted ethylene glycol group having 2 to 20 carbon atoms, a substituted or unsubstituted alcohol group having 1 to 20 carbon atoms, a substituted or unsubstituted alkyl halogen group having 1 to 20 carbon atoms, a substituted or unsubstituted thiol group having 1 to 20 carbon atoms, a substituted or unsubstituted alkyl azide group having 3 to 20 carbon atoms, a substituted or unsubstituted aryl azide group having 7 to 30 carbon atoms, a substituted or unsubstituted alkenyl group having 2 to 40 carbon atoms, a substituted or unsubstituted alkynyl group having 2 to 40 carbon atoms, a cyano group, a halogen group, deuterium and hydrogen; L H1 , L H3 and L H4 in Formula 4 combine with each other around L H2 to form a tetradentate ligand represented by Formula 7 below; In this case, L H5 and L H6 each form a monodentate ligand for the transition metal M as represented by Chemical Formula 8; or L H5 and L H6 each combine with each other to form a bidentate ligand for the transition metal M as represented by Chemical Formula 9 below; [Chemical formula 7] 【Chemistry 7】 In the above Chemical Formula 7, C a , C b , and C c are each independently a heterocyclic compound containing one or more nitrogen atoms, the heterocyclic compound is connected to an amine group at the 2-position by a methylene group, three nitrogen atoms in the three heterocyclic rings and one central nitrogen atom connecting the three heterocyclic rings are connected to a transition metal M, R 1 , R 2 , and R 3 are each a functional group of the heterocyclic compound C a , C b , and C c , the R 1 , R 2 , and R 3 are each independently selected from the group consisting of a substituted or unsubstituted alkyl group having 1 to 10 carbon atoms, a substituted or unsubstituted ethylene glycol group having 2 to 20 carbon atoms, a substituted or unsubstituted alcohol group having 1 to 20 carbon atoms, a substituted or unsubstituted alkyl halogen group having 1 to 20 carbon atoms, a substituted or unsubstituted thiol group having 1 to 20 carbon atoms, a substituted or unsubstituted alkyl azide group having 3 to 20 carbon atoms, a substituted or unsubstituted aryl azide group having 7 to 30 carbon atoms, a substituted or unsubstituted alkenyl group having 2 to 40 carbon atoms, a substituted or unsubstituted alkynyl group having 2 to 40 carbon atoms, a cyano group, a halogen group, deuterium, and hydrogen; [Chemical formula 8] 【Chemistry 8】 [Chemical formula 9] 【Chemistry 9】 In the above Chemical Formula 8, L H5 and L H6 are each a monodentate ligand, and each independently is —H, —F, —Cl, —Br, —I, —NO 2 , —NCCH 3 , —CO, —OH 2 , —NH 3 or a heterocyclic compound containing one or more nitrogen atoms; In the above Chemical Formula 9, L H5 -L H6 is a bidentate ligand and is catechol, acetylacetone, 2-picolinic acid, 2-pyridinecarboxamide, 2,2-bipyridine, or 2,2-bithiazole; L 1 is independently selected from the group consisting of a substituted or unsubstituted alkylene group having 1 to 20 carbon atoms, a substituted or unsubstituted cycloalkylene group having 1 to 20 carbon atoms, a substituted or unsubstituted ethylene glycol group having 2 to 30 carbon atoms, a substituted or unsubstituted arylene group having 6 to 30 carbon atoms, and a substituted or unsubstituted heteroarylene group having 3 to 30 carbon atoms; A a is selected from the group consisting of an amine group and an ammonium group.

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

3. A method for producing a thin film of an oxidation-reduction polymer material for an electrochemical sensor, comprising the steps of coating an electrode with the oxidation-reduction polymer material for an electrochemical sensor according to claim 1, and then curing the coated electrode.

4. an enzyme capable of oxidizing or reducing a liquid biological sample; and A sensing membrane for an electrochemical biosensor, comprising the oxidation-reduction polymer material for an electrochemical sensor according to claim 1.

5. The enzyme is one or more oxidoreductases selected from the group consisting of dehydrogenases, oxidases, and esterases; or 5. The sensing membrane for an electrochemical biosensor according to claim 4, comprising 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).

6. A continuous blood glucose monitoring sensor comprising the sensing membrane for an electrochemical biosensor according to claim 4.

7. A continuous blood glucose monitoring device comprising the oxidation-reduction polymer material for an electrochemical sensor according to claim 1 or 2.

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