Electrochemical biosensors or sensing films for electrochemical biosensors containing transition metal complexes or redox polymers

Abstract

The present invention relates to a transition metal complex having a bidentate ligand including pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole, which is used as an electron transfer mediator in a continuous blood glucose meter for measuring blood glucose concentration, and an oxidation-reduction polymer containing the same. The transition metal complex and the oxidation-reduction polymer can rapidly and smoothly exchange electrons between an enzyme and an electrode, and are therefore useful for a continuous blood glucose biosensor.

Description

[Technical Field]

[0001] The present invention relates to a transition metal complex having a bidentate ligand, such as pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole, and an electrochemical biosensor utilizing the same. [Background technology]

[0002] Recently, interest in the development of biosensors for quantitative and qualitative analysis of target analytes has been increasing day by day in fields ranging from medicine to the environment and food. In particular, enzyme-based biosensors are chemical sensors used to selectively detect and measure chemical substances contained in a sample by utilizing the biological sensing function of living organisms, such as functional substances in living organisms or microorganisms, which sensitively react to specific substances. They have been developed primarily for medical measurement applications such as blood glucose sensors, and active research is also being conducted into their applications in food engineering and environmental measurement.

[0003] Periodic measurement of blood glucose is very important in diabetes management, and therefore, various blood glucose monitors have been developed to allow easy measurement of blood glucose using portable measuring devices. The operating principle of such biosensors is based on optical or electrochemical methods. Unlike conventional optical biosensors, electrochemical biosensors have the advantages of being able to reduce the influence of oxygen and allowing samples to be used without additional pretreatment even if they are turbid. Therefore, various types of electrochemical biosensors with high accuracy and precision are widely used.

[0004] Currently available electrochemical blood glucose sensors mainly use enzyme electrodes. Specifically, they have a structure in which glucose oxidase is immobilized chemically or physically on an electrode that can convert glucose into an electrical signal. These electrochemical blood glucose sensors measure the concentration of glucose in an analyte, such as blood, by transferring electrons generated when glucose in the analyte is oxidized by an enzyme to the electrode and measuring the resulting current. Biosensors that use enzyme electrodes have the problem that the distance from the active center of the enzyme is too great to directly transfer electrons generated by the oxidation of the substrate to the electrode. Therefore, a redox mediator, i.e., an electron transfer mediator, is required to facilitate this electron transfer reaction. Therefore, the type of enzyme used and the characteristics of the electron transfer mediator are the most important factors that determine the performance of electrochemical biosensors for measuring blood glucose.

[0005] The development of blood glucose sensors has progressed to the point where, in order to prevent changes in measurement values ​​due to differences in oxygen partial pressure (pO2) that vary depending on the blood (venous blood, capillary blood, etc.), there has been a shift to using GDH, in which oxygen is excluded from the enzymatic reaction, instead of GOX, in which oxygen participates in the enzymatic reaction with glucose in the blood.In the case of electron transfer mediators, ferricyanide, which is sensitive to stability due to humidity, has been replaced by organic compounds such as quinone derivatives (phenanthroline quinone, quineonediimine, etc.), which are highly stable under temperature and humidity, and organometallic compounds such as Ru complexes (ruthenium hexamine, etc.) and osmium complexes.

[0006] The most commonly used electron transfer mediator is potassium ferricyanide [K3Fe(CN)6], which is inexpensive, highly reactive, and useful for sensors using FAD-GOX, PQQ-GDH, or FAD-GDH. However, sensors using this electron transfer mediator require special care in manufacturing and storage because they are susceptible to measurement errors due to interfering substances such as uric acid and gentisic acid present in blood and are susceptible to deterioration due to temperature and humidity. Furthermore, after long-term storage, changes in background current make it difficult to accurately detect low concentrations of glucose.

[0007] Hexaammineruthenium chloride [Ru(NH3)6Cl3] has higher redox stability than ferricyanide, and biosensors using this electron transfer mediator are easy to manufacture and store, and have the advantage of being highly stable with little change in background current even when stored for long periods of time. However, they have the disadvantage of being incompatible with FAD-GDH in terms of reactivity, making them difficult to manufacture into commercially useful sensors.

[0008] Furthermore, when using such biosensors, obtaining accurate and fast measurements with a small amount of sample is a very important issue in order to maximize user convenience.

[0009] Therefore, there is still a demand for the development of new electron transfer mediators that can overcome the shortcomings of conventional electron transfer mediators and achieve a reduction in measurement time.

[0010] Continuous glucose monitoring (CGM) systems are used to continuously monitor blood glucose levels and manage diseases such as diabetes. However, conventional enzyme sensors, which collect blood from the fingertip, are not used in CGM because the needle causes considerable pain during blood collection, limiting the frequency of measurements. To address this issue, improved enzyme sensors that can be attached to the body and minimize invasiveness have recently been developed. In the case of enzyme sensors for continuous glucose monitoring, part of the sensor is inserted into the human body, and electron transfer mediators containing transition metals, as mentioned above, are immobilized with polymers such as polyvinylpyridine or polyvinylimidazole to prevent absorption by the human body and cause toxicity and side effects, thereby preventing problems caused by the loss of electron transfer mediators within the body.

[0011] Thus, in order to develop a novel electron transfer mediator suitable for CGMS sensors, transition metal electron transfer mediators containing bipyridine and bisimidazole ligands have been immobilized onto polymer backbones to prepare and use oxidation-reduction polymers for enzyme sensors. Under these circumstances, the present inventors have confirmed that all of these requirements can be met by using transition metal complexes with bidentate ligands containing heterocyclic compounds such as pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole in addition to bipyridine and bisimidazole ligands, and have thus completed the present invention. Summary of the Invention [Problem to be solved by the invention]

[0012] The present invention provides a transition metal complex or a salt compound thereof having a bidentate ligand containing a heterocyclic structure such as pyrazole, triazole, tetrazole, oxadiazole or thiadiazole, which allows for easy synthesis of various derivatives and introduction of functional groups.

[0013] As an example, the transition metal complex may be a transition metal complex or a salt compound thereof having a bidentate ligand containing one structure selected from the group consisting of pyridine; and pyrazole, triazole, tetrazole, oxadiazole, and thiadiazole.

[0014] Another object of the present invention is to provide an oxidation-reduction polymer containing the transition metal complex or a salt compound thereof.

[0015] It is still another object of the present invention to provide a device comprising the transition metal complex or a salt compound thereof and / or an oxidation-reduction polymer containing the same.

[0016] In one example, the device may be an electron transfer mediator device, specifically an electrochemical biosensor. In one example, the device may be insertable into the body. In one example, the electrochemical biosensor may be a blood glucose sensor.

[0017] Yet another object of the present invention is to provide a sensing membrane for an electrochemical biosensor, which comprises an enzyme capable of oxidizing-reducing a liquid biological sample; and the transition metal complex or its salt compound and / or an oxidation-reduction polymer containing the same. [Means for solving the problem]

[0018] In one aspect, the present invention provides a transition metal complex or a salt compound thereof having a bidentate (hereinafter also referred to as "bidentate ligand") ligand containing one structure selected from the group consisting of pyridine, and pyrazole, triazole, tetrazole, oxadiazole, and thiadiazole.

[0019] Specifically, the transition metal complex may be a compound represented by the following Chemical Formula 1: [Chemical formula 1] [M(L)a (X1) b ] c d(X2) In the above formula, M is a transition metal selected from the group consisting of Fe, Ru, and Os; L is a bidentate ligand comprising one structure selected from the group consisting of pyridine; and pyrazole, triazole, tetrazole, oxadiazole, and thiadiazole; a is 2 or 3, X1 is one halogen atom selected from the group consisting of F, Cl, Br, and I; b is 0, 1 or 2; c is an integer selected from 1 to 3 (e.g., 1, 2, or 3); X2 is a counter ion selected from the group consisting of F, Cl, Br, I, and PF6; d is 0, 1 or 2.

[0020] The present invention will be described in detail below.

[0021] Unless otherwise defined, all technical terms used herein are used in the sense commonly understood by those skilled in the art in the field related to the present invention. Furthermore, while preferred methods and samples are described herein, similar or equivalent methods and samples are also included within the scope of the present invention. Furthermore, numerical values ​​described herein are considered to include the meaning of "about" even if not explicitly stated. The contents of all publications referenced herein are incorporated herein by reference.

[0022] The definitions of residues used herein are explained in detail below. Unless otherwise specified, each residue has the following definition and is used in the sense that it is commonly understood by a person of ordinary skill in the art.

[0023] As used herein, "halo" or "halogen" refers to, for example, fluorine, chlorine, bromine and iodine.

[0024] As used herein, "alkyl" refers to an aliphatic hydrocarbon group, including all straight-chain and branched hydrocarbon groups. For example, aliphatic hydrocarbons having 1 to 6 carbon atoms include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, 1-ethylpropyl, hexyl, isohexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, and 2-ethylbutyl. Unless otherwise defined, alkyl refers to alkyls having 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, 1 to 2 carbon atoms, 2 to 6 carbon atoms, 2 to 5 carbon atoms, 2 to 4 carbon atoms, 2 to 3 carbon atoms, 3 to 6 carbon atoms, 3 to 5 carbon atoms, 3 to 4 carbon atoms, 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms.

[0025] As used herein, "alkoxy" refers to an -O-alkyl or alkyl-O- group, where the alkyl group is as defined above. Examples include, but are not limited to, methoxy, ethoxy, n-propoxy, n-butoxy, t-butoxy, etc. The alkoxy group can be substituted or unsubstituted with one or more suitable groups.

[0026] As used herein, the terms "hydroxy" or "hydroxyl," alone or in combination with other terms, means --OH.

[0027] As used herein, "amino" refers to -NH2 and "nitro" refers to -NO2.

[0028] As used herein, "aryl" refers to a monovalent aromatic hydrocarbon derived by removing a hydrogen atom from a carbon atom in a parent aromatic ring system, e.g., having 6 to 20 carbon atoms, 6 to 12 carbon atoms, or 6 to 10 carbon atoms. The aryl can include bicyclic radicals containing an aromatic ring fused with a saturated or partially unsaturated ring.

[0029] Examples of aryl groups include, but are not limited to, radicals derived from benzene (phenyl), substituted phenyl, biphenyl, naphthyl, tetrahydronaphthyl, fluorenyl, toluyl, naphthalenyl, anthracenyl, indenyl, indanyl, etc. Aryl groups can be substituted or unsubstituted with one or more suitable groups.

[0030] In this specification, unless otherwise specified in the present invention, "substituted" means that at least one hydrogen atom has been 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, a hydroxyl group having 2 to 6 carbon atoms, a hydroxyl ... alkenyl groups having 2 to 6 carbon atoms, haloalkenyl groups having 2 to 6 carbon atoms, alkynyl groups having 2 to 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, cycloalkyl groups having 3 to 20 carbon atoms, 5- to 12-membered heterocycloalkyl groups, 5- to 12-membered heteroaryl groups, aryl groups having 6 to 10 carbon atoms, aryloxy groups having 6 to 10 carbon atoms, and arylthio groups having 6 to 10 carbon atoms.

[0031] The transition metal complexes provided herein can be compounds of Formula 1: [Chemical formula 1] [M(L) a (X1) b ] c d(X2) In the above formula, M is a transition metal selected from the group consisting of Fe, Ru, and Os; L is a bidentate ligand comprising one structure selected from the group consisting of pyridine; and pyrazole, triazole, tetrazole, oxadiazole, and thiadiazole; a is 2 or 3, X1 is one halogen atom selected from the group consisting of F, Cl, Br, and I; b is 0, 1 or 2; c is an integer selected from 1 to 3 (e.g., 1, 2, or 3); X2 is a counter ion selected from the group consisting of F, Cl, Br, I, and PF6; d is 0, 1 or 2.

[0032] The pyridine is unsubstituted or 1-4 Alkyl group, C 1-4 Alkoxy group, -(CH2)-OC 1-4 Alkyl group, -(CH2CH2)-OC 1-4 Alkyl groups, and C 1-4 It may be substituted with one or more (for example, 1 to 4, 1, 2, 3, or 4) alkylamino groups selected from the group consisting of alkylamino groups.

[0033] The pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole is each unsubstituted or 1-4 Alkyl group, C 1-4 Alkoxy group, -(CH2)-OC 1-4 Alkyl group, -(CH2CH2)-OC 1-4 alkyl groups, [ka] , and C 1-4 It may be substituted with one or more (for example, 1 to 3, 1, 2, or 3) alkylamino groups selected from the group consisting of alkylamino groups.

[0034] The R'4 is hydrogen or substituted or unsubstituted C 1-4 alkyl, wherein n' is an integer selected from 1 to 4, for example, 1, 2, 3, or 4.

[0035] Said C 1-4The alkyl group, alkoxy group, or alkylamino group means an alkyl group, alkoxy group, or alkylamino group having 1 to 4, 1 to 3, 1 to 2, 2 to 4, 2 to 3, 3 to 4, 1, 2, 3, or 4 carbon atoms.

[0036] Said C 1-4 Alkyl group, C 1-4 Alkoxy group, -(CH2)-OC 1-4 Alkyl group, -(CH2CH2)-OC 1-4 Alkyl group, or C 1-4 The alkylamino group can be unsubstituted or substituted. 1-4 Alkyl group, C 1-4 Alkoxy group, -(CH2)-OC 1-4 Alkyl group, -(CH2CH2)-OC 1-4 Alkyl group, or C 1-4 In the alkylamino group, the hydrogen atom may be substituted with a halogen atom such as F, Cl, Br, or I, a cyano group, a hydroxyl group, a thiol group, a nitro group, an amino group, an imino group, an azido 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, or a phosphoric acid group or a salt thereof.

[0037] Specifically, the transition metal complex may be a compound represented by the following formula 2: [Chemical formula 2] [ka] In the above formula, R1, R2, R3 and R4 are each independently hydrogen, C 1-4 Alkyl group, C 1-4 Alkoxy group, -(CH2)-OC 1-4 Alkyl group, -(CH2CH2)-OC 1-4 Alkyl group, or C 1-4 is an alkylamino group, n is 0, At least one of W', Y', Z', and V' is nitrogen (N); W' is nitrogen (N) or carbon (C), Y', Z', and V' are each independently nitrogen (N), sulfur (S), oxygen (O), or carbon (C); R'1, R'2, and R'3 are each independently hydrogen, C 1-4 Alkyl group, C 1-4 Alkoxy group, -(CH2)-OC 1-4 Alkyl group, -(CH2CH2)-OC 1-4 alkyl groups, [ka] , or C 1-4 is an alkylamino group, The dotted line represents a bond or no bond, The R'4 is hydrogen or substituted or unsubstituted C 1-4 is alkyl, n' is an integer selected from 1 to 4, for example, 1, 2, 3, or 4; M, a, X1, b, c, X2 and d are as defined in Chemical Formula 1 above.

[0038] Said C 1-4 Alkyl group, C 1-4 Alkoxy group, -(CH2)-OC 1-4 Alkyl group, -(CH2CH2)-OC 1-4 Alkyl group, or C 1-4 Each alkylamino group can be unsubstituted or substituted, and if substituted, is as described above.

[0039] Specifically, the transition metal complex may be a compound selected from the following Chemical Formula 3 to Chemical Formula 25. [Chemical formula 3] [ka] [Chemical formula 4] [ka] [Chemical formula 5]

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[0040] For example, the transition metal complex according to the present invention may include an oxidized transition metal complex, specifically a trivalent osmium complex or a divalent osmium complex. The oxidizing agent used in the oxidation treatment may be any commonly used oxidizing agent, such as NaOCl, HO, O, O, PbO, MnO, KMnO, ClO, F, Cl, HCrO, NO, AgO, OsO, HSO, ceric ammonium nitrate (CAN), pyridinium chlorochromate, and 2,2'-dipyridyldisulfide. Furthermore, when the transition metal complex includes a compound in both an oxidized and reduced state, the oxidized transition metal complex or its salt compound can be obtained by oxidation treatment.

[0041] The transition metal complex according to the present invention is in the form of a salt compound having a suitable counter ion and / or ion, and the salt compound may have high solubility in water, an aqueous solution or an organic solvent. - , Cl - and Br - When composed of small opposite anions such as I, they tend to be highly soluble in water or aqueous solutions. - , hexafluorophosphate (PF6 - ) and tetrafluoroborate (BF4 - ), they tend to be highly soluble in organic solvents. Examples of the counter anion may be one or more of halides selected from the group consisting of F, Cl, Br, and I, hexafluorophosphate, and tetrafluoroborate.

[0042] In another aspect, the present invention provides an oxidation-reduction polymer comprising the transition metal complex or a salt compound thereof and including a polymer backbone such as polyvinylimidazole (PVI) and polyvinylpyridine (PVP).

[0043] Specifically, the oxidation-reduction polymer may be a compound of the following formula 26 or 27: [Chemical formula 26] [ka] [Chemical formula 27] [ka] In the above formula, M is a transition metal selected from the group consisting of Fe, Ru, and Os; L is a bidentate ligand comprising one structure selected from the group consisting of pyridine; and pyrazole, triazole, tetrazole, oxadiazole, and thiadiazole; a is 2 or 3, X1 is one halogen atom selected from the group consisting of F, Cl, Br, and I; X2 is a counter ion selected from the group consisting of F, Cl, Br, I, and PF6; m and o are each an integer selected from 10 to 600.

[0044] The pyridine is unsubstituted or 1-4 Alkyl group, C 1-4 Alkoxy group, -(CH2)-OC 1-4 Alkyl group, -(CH2CH2)-OC 1-4 Alkyl groups, and C 1-4 It may be substituted with one or more (for example, one, two, three, or four) alkylamino groups selected from the group consisting of alkylamino groups.

[0045] The pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole is each unsubstituted or 1-4 Alkyl group, C 1-4 Alkoxy group, -(CH2)-OC 1-4 Alkyl group, -(CH2CH2)-OC 1-4 alkyl groups, [ka] , and C 1-4 It may be substituted with one or more (for example, one, two, or three) alkylamino groups selected from the group consisting of alkylamino groups.

[0046] Specifically, the oxidation-reduction polymer may be a compound selected from the following formulas 28 to 45: [Chemical formula 28] [ka] [Chemical formula 29] [ka] [Chemical formula 30]

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[0047] In yet another aspect of the present invention, the oxidation-reduction polymer further comprises a crosslinkable functional group and may be a compound of Formula 46 or 47 below. [Chemical formula 46] [ka] [Chemical formula 47] [ka] In the above formula, M is a transition metal selected from the group consisting of Fe, Co, Ru, Os, Rh, and Ir; L is a bidentate ligand comprising one structure selected from the group consisting of pyridine; and pyrazole, triazole, tetrazole, oxadiazole, and thiadiazole; a is 2 or 3, X1 is one halogen atom selected from the group consisting of F, Cl, Br, and I; X2 is a counter ion selected from the group consisting of F, Cl, Br, I, and PF6; A D are primary and secondary amine groups, ammonium groups, halogen groups, epoxy groups, azide groups, acrylate groups, alkenyl groups, alkynyl groups, thiol groups, isocyanates, alcohol groups, silane groups, and [ka] is one selected from the group consisting of The R5' is hydrogen or substituted or unsubstituted C 1-4 is alkyl, wherein n'' is an integer selected from 1 to 4, q is an integer selected from 1 to 10; m, o, and p are each an integer selected from 10 to 600.

[0048] The pyridine is unsubstituted or 1-4 Alkyl group, C 1-4 Alkoxy group, -(CH2)-OC 1-4 Alkyl group, -(CH2CH2)-OC 1-4 Alkyl groups, and C 1-4 It may be substituted with one or more (for example, one, two, three, or four) alkylamino groups selected from the group consisting of alkylamino groups.

[0049] The pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole is each unsubstituted or 1-4 Alkyl group, C 1-4 Alkoxy group, -(CH2)-OC 1-4 Alkyl group, -(CH2CH2)-OC 1-4 alkyl groups, [ka] , and C 1-4 It may be substituted with one or more (for example, one, two, or three) alkylamino groups selected from the group consisting of alkylamino groups.

[0050] Specifically, the oxidation-reduction polymer may be a compound selected from the following formulas 48 to 60. [Chemical formula 48] [ka] [Chemical formula 49] [ka] [Chemical formula 50] [ka] [Chemical formula 51] [ka] [Chemical formula 52] [ka] [Chemical formula 53] [ka] [Chemical formula 54] [ka] [Chemical formula 55] [ka] [Chemical formula 56] [ka] [Chemical formula 57] [ka] [Chemical formula 58] [ka] [Chemical formula 59] [ka] and [Chemical formula 60] [ka]

[0051] Yet another aspect provides a device comprising the transition metal complex or a salt compound thereof; or the oxidation-reduction polymer.

[0052] As an example, the device may be an electrochemical biosensor.

[0053] As an example, the device may be insertable into the human body, and in particular may be an electrochemical biosensor insertable into the human body.

[0054] As an example, the electrochemical biosensor can be a blood glucose sensor, for example, an electrochemical glucose (blood sugar) sensor.

[0055] As an example, the electrochemical biosensor can be a continuous glucose monitoring sensor.

[0056] The continuous blood 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, a protection layer, etc. In the case of electrodes, the sensor 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.

[0057] As an example, the biosensor according to the present invention may be an electrochemical biosensor fabricated by applying a reagent composition containing the transition metal complex or its salt compound, or the oxidation-reduction polymer and an enzyme capable of oxidizing and reducing a liquid biological sample, to a substrate having at least two, preferably two or three, electrodes, and then drying the applied composition.

[0058] 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 film containing the transition metal complex or redox polymer of the present invention is laminated on the working electrode, and an insulator, a diffusion film, and a protective film are laminated in this order on both sides of the substrate on which the working electrode and the counter electrode are provided.

[0059] 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).

[0060] The working electrode may be a carbon, gold, platinum, silver, or silver / silver chloride electrode.

[0061] 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.

[0062] 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 a polyurethane-based copolymer, but is not limited to these.

[0063] 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.

[0064] By varying 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, alcohol, amino acids, and glutamate.

[0065] In yet another embodiment, there is provided a sensing membrane for an electrochemical biosensor, comprising an enzyme capable of oxidizing-reducing a liquid biological sample; and the transition metal complex or a salt compound thereof; or the oxidation-reduction polymer as an electron transfer mediator.

[0066] The liquid biological sample may be, for example, but is not limited to, one or more, two or more, three or more, four or more, or five or more selected from the group consisting of tissue fluid, blood, cells, plasma, serum, urine, cyst fluid, and saliva of a patient.

[0067] In one example, the enzyme is one or more oxidoreductases selected from the group consisting of dehydrogenases, oxidases, and esterases; or The enzyme may comprise 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).

[0068] 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 the signal generated, such as a change in current. 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.

[0069] 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.

[0070] Meanwhile, the oxidoreductase may contain a cofactor that serves to store the hydrogen removed by the oxidoreductase from the target substance (e.g., the 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.

[0071] For example, when measuring blood glucose concentration, 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.

[0072] 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.

[0073] 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.

[0074] 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.

[0075] 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.

[0076] The sensing membrane according to the present invention may further include a cross-linking agent.

[0077] 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.

[0078] The surfactant may serve to ensure that the composition is evenly spread and dispensed at a uniform thickness 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 composition is evenly spread and dispensed at a uniform thickness 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.

[0079] The water-soluble polymer may function as a polymer support for the reagent composition, stabilizing and dispersing 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.

[0080] 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.

[0081] 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 ensure that the oxidation-reduction polymer according to the present invention firmly attaches 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. [Effects of the Invention]

[0082] The transition metal complexes and redox polymers according to the present invention can easily adjust the potential value depending on the type of ligand introduced. Compared to conventional bipyridine systems, the size of the ligand is smaller and the electron transfer rate is increased, so electrochemical biosensors using these have the advantages of rapid detection and economical cost. [Brief explanation of the drawings]

[0083] [Figure 1a] 1A is a cyclic voltammetry curve showing the electrochemical properties of a transition metal complex having a bidentate ligand containing pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention {Chemical Formula 3 (FIG. 1a)}. [Figure 1b] 1B is a cyclic voltammetry curve showing the electrochemical properties of a transition metal complex having a bidentate ligand containing pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention. {Chemical Formula 4 (FIG. 1B)} [Figure 1c]1c is a cyclic voltammetry curve showing the electrochemical properties of a transition metal complex having a bidentate ligand containing pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention. [Figure 1d] 1d is a cyclic voltammetry curve showing the electrochemical properties of a transition metal complex having a bidentate ligand containing pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention. [Figure 1e] 1 is a cyclic voltammetry curve showing the electrochemical properties of a transition metal complex having a bidentate ligand containing pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention. {Chemical Formula 14 (FIG. 1e)} [Figure 1f] 1f shows cyclic voltammetry curves illustrating the electrochemical properties of transition metal complexes having bidentate ligands including pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention. [Figure 1g] 1g shows cyclic voltammetry curves illustrating the electrochemical properties of transition metal complexes having bidentate ligands including pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention. [Figure 1h] 1 is a cyclic voltammetry curve showing the electrochemical properties of a transition metal complex having a bidentate ligand containing pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention {Chemical Formula 17 (FIG. 1h)}. [Figure 1i] 1 is a cyclic voltammetry curve showing the electrochemical properties of a transition metal complex having a bidentate ligand containing pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention {Chemical Formula 18 (FIG. 1i)}. [Figure 1j]1 is a cyclic voltammetry curve showing the electrochemical properties of a transition metal complex having a bidentate ligand containing pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention. {Chemical Formula 20 (FIG. 1j)} [Figure 1k] 1 is a cyclic voltammetry curve showing the electrochemical properties of a transition metal complex having a bidentate ligand containing pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention. {Chemical Formula 22 (FIG. 1k)} [Figure 1l] 11 is a cyclic voltammetry curve showing the electrochemical properties of a transition metal complex having a bidentate ligand containing pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention. {Chemical Formula 23 (FIG. 1l)} [Figure 1m] 1m is a cyclic voltammetry curve showing the electrochemical properties of a transition metal complex having a bidentate ligand containing pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention. [Figure 1n] 1 is a cyclic voltammetry curve showing the electrochemical properties of a transition metal complex having a bidentate ligand containing pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention {Chemical Formula 25 (FIG. 1n)}. [Figure 1o] 10 shows cyclic voltammetry curves illustrating the electrochemical properties of transition metal complexes having bidentate ligands including pyrazole, triazole, tetrazole, oxadiazole, or thiadiazole according to the present invention (Chemical Formulas 3, 4, 11, 14, 15, and 16 (FIG. 1o)). [Figure 2] 1 is a cyclic voltammetry curve showing the electrochemical properties of a redox polymer containing a transition metal complex according to the present invention. [Figure 3] 1 is a cyclic voltammetry curve showing the electrochemical properties of a transition metal complex and an oxidation-reduction polymer comprising a crosslinkable functional group according to the present invention. [Figure 4] FIG. 1 is a diagram showing the potential of an electrode to which an oxidation-reduction polymer according to the present invention is applied. [Figure 5] 1 is a graph showing that all electrodes using the oxidation-reduction polymer according to the present invention exhibit linearity with respect to glucose concentrations of 10 mM or less, and show similar sensitivity to the control electrodes despite the application of a lower voltage. [Figure 6] 1 is a graph showing that all electrodes using the oxidation-reduction polymer according to the present invention exhibit linearity with respect to glucose concentrations of 10 mM or less, and show similar sensitivity to the control electrodes despite the application of a lower voltage. [Figure 7] 1 is a graph showing that all electrodes to which the oxidation-reduction polymer according to the present invention is applied respond to glucose at a lower voltage than the control electrodes. DETAILED DESCRIPTION OF THE INVENTION

[0084] 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.

[0085] Example 1: Preparation of transition metal complexes according to the present invention Example 1.1. Synthesis of transition metal complexes of formula 3 1) Synthesis of 2-(1H-pyrazol-1-yl)pyridine [ka]

[0086] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 4.7 g (69 mmol) of pyrazole and 9.3 g (83 mmol) of potassium tert-butoxide, dissolved in 40 mL of anhydrous dimethyl sulfoxide under an argon atmosphere. To this reaction mixture, 8.0 g (83 mmol) of 2-fluoropyridine was added and heated to 100 °C under an argon atmosphere and stirred for 4 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was dried over magnesium sulfate and concentrated under reduced pressure to give a clear, colorless solid (7.2 g, 72%).

[0087] 2) Synthesis of Os(pzpy)2Cl2 [chemical formula 3] [ka] 5.0 g (10 mmol) of potassium hexachloroosmate(IV) and 2.9 g (20 mmol) of 2-(1H-pyrazol-1-yl)pyridine prepared in 1) were placed in a 500 mL Schlenk flask and dissolved in 200 mL of ethylene glycol under an argon gas atmosphere. The mixture was then degassed with argon for 15 minutes. The reaction mixture was heated to 180°C and stirred for 1 hour. After the reaction was completed, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to a 1.0 M aqueous solution of sodium dithionite (250 mL) to obtain a reduced osmium complex precipitate. The resulting solid was filtered under vacuum, washed several times with water, and dried in a vacuum oven at 40°C to obtain the final green osmium complex (4.0 g, 75%). HRMS ( 192 Os): m / z 552.0240 ([M + ]required 552.0261)

[0088] The overall method for preparing the compound of Chemical Formula 3 is shown in Reaction Scheme 1 below. [Reaction Scheme 1] [ka]

[0089] Example 1.2. Synthesis of transition metal complexes of formula 4 1) Synthesis of 2-methyl-6-(1H-pyrazol-1-yl)pyridine [ka]

[0090] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 2.0 g (30 mmol) of pyrazole and 4.0 g (36 mmol) of potassium tert-butoxide, dissolved in 20 mL of anhydrous dimethyl sulfoxide under an argon atmosphere. 5.0 g (36 mmol) of 2-fluoro-6-methylpyridine was added to the reaction mixture, which was heated to 100 °C under argon atmosphere and stirred for 4 h. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was concentrated under reduced pressure to remove the solvent, and then purified by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 5:1). Finally, a transparent solid, 2-methyl-6-(1H-pyrazol-1-yl)pyridine, was obtained (1.4 g, 30%).

[0091] 2) Synthesis of Os(pz-2-Me-py)2Cl2 [chemical formula 4] [ka]

[0092] In a 250 mL Schlenk flask, 1.5 g (3.1 mmol) of potassium hexachloroosmate(IV) and 1.0 g (6.3 mmol) of 2-methyl-6-(1H-pyrazol-1-yl)pyridine prepared in 1) above were placed and dissolved in 50 mL of ethylene glycol under an argon gas atmosphere. The mixture was then degassed with argon for 15 minutes. The reaction mixture was heated to 180°C and stirred for 1 hour. After the reaction was completed, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to a 1.0 M aqueous solution of sodium dithionite (250 mL) to obtain a reduced osmium complex precipitate. The resulting solid was filtered under vacuum, washed multiple times with water and acetonitrile, and then dried in a vacuum oven to obtain the final green osmium complex (0.27 g, 15%). HRMS ( 192 Os): m / z 580.0569 ([M + ]required 580.0574)

[0093] The overall method for producing the compound of Chemical Formula 4 is shown in Reaction Scheme 2 below. [Reaction Scheme 2] [ka]

[0094] Example 1.3. Synthesis of transition metal complexes of formula 5 1) Synthesis of 4-methoxy-2-(1H-pyrazol-1-yl)pyridine [ka] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 2.0 g (30 mmol) of pyrazole and 4.0 g (36 mmol) of potassium tert-butoxide, dissolved in 20 mL of anhydrous dimethyl sulfoxide under an argon atmosphere. 6.7 g (36 mmol) of 2-bromo-4-methoxypyridine was added to the reaction mixture, which was heated to 100 °C under an argon atmosphere and stirred for 8 h. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was concentrated under reduced pressure to remove the solvent, and then purified by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 3:1). Finally, a transparent solid, 4-methoxy-2-(1H-pyrazol-1-yl)pyridine, was obtained (4.0 g, 63%).

[0095] 2) Synthesis of Os(pz-4-Meo-py)2Cl2 [Chemical Formula 5] [ka] 2.0 g (4.2 mmol) of potassium hexachloroosmate(IV) and 1.5 g (8.3 mmol) of 4-methoxy-2-(1H-pyrazol-1-yl)pyridine prepared in 1) above were placed in a 250 mL Schlenk flask and dissolved in 50 mL of ethylene glycol under an argon gas atmosphere. The mixture was then degassed with argon for 15 minutes. The reaction mixture was heated to 180°C and stirred for 1 hour. After the reaction was completed, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to 250 mL of a 1.0 M aqueous solution of sodium dithionite to obtain a reduced osmium complex precipitate. The resulting solid was filtered under vacuum, washed multiple times with water and acetonitrile, and dried in a vacuum oven to obtain the final green osmium complex (2.0 g, 78%). HRMS ( 192 Os): m / z 612.0460 ([M + ]required 612.0472)

[0096] The overall method for preparing the compound of Chemical Formula 5 is shown in Reaction Scheme 3 below. [Reaction Scheme 3] [ka]

[0097] Example 1.4. Synthesis of transition metal complexes of formula 6 1) Synthesis of 4-methyl-2-(1H-pyrazol-1-yl)pyridine [ka] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 2.0 g (30 mmol) of pyrazole and 4.0 g (36 mmol) of potassium tert-butoxide, dissolved in 20 mL of anhydrous dimethyl sulfoxide under an argon atmosphere. 6.2 g (36 mmol) of 2-bromo-4-methylpyridine was added to the reaction mixture, which was heated to 100 °C under an argon atmosphere and stirred for 8 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was concentrated under reduced pressure to remove the solvent, followed by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 3:1). Finally, a transparent solid, 4-methoxy-2-(1H-pyrazol-1-yl)pyridine, was obtained (3.5 g, 61%).

[0098] 2) Synthesis of Os(pz-4-Me-py)2Cl2 [Chemical Formula 6] [ka] 2.0 g (4.2 mmol) of potassium hexachloroosmate(IV) and 1.3 g (8.3 mmol) of 4-methyl-2-(1H-pyrazol-1-yl)pyridine prepared in 1) above were placed in a 250 mL Schlenk flask and dissolved in 50 mL of ethylene glycol under an argon gas atmosphere. The mixture was then degassed with argon for 15 minutes. The reaction mixture was heated to 180°C and stirred for 1 hour. After the reaction was completed, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to a 1.0 M aqueous solution of sodium dithionite (250 mL) to obtain a reduced osmium complex precipitate. The resulting solid was filtered under vacuum, washed multiple times with water and acetonitrile, and dried in a vacuum oven to obtain the final green osmium complex (1.0 g, 42%). HRMS ( 192 Os): m / z 580.0561 ([M + ]required 580.0574)

[0099] The overall method for preparing the compound of Chemical Formula 6 is shown in Reaction Scheme 4 below. [Reaction Scheme 4] [ka]

[0100] Example 1.5. Synthesis of transition metal complex of formula 7 1) Synthesis of 4-methyl-2-(3-methyl-1H-pyrazol-1-yl)pyridine [ka] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 2.5 g (30 mmol) of 3-methylpyrazole and 4.0 g (36 mmol) of potassium tert-butoxide, dissolved in 20 mL of anhydrous dimethyl sulfoxide under an argon atmosphere. 6.2 g (36 mmol) of 2-bromo-4-methylpyridine was added to the reaction mixture, which was heated to 100 °C under argon atmosphere and stirred for 18 h. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was concentrated under reduced pressure to remove the solvent, followed by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 3:1). Finally, a transparent solid, 4-methyl-2-(3-methyl-1H-pyrazol-1-yl)pyridine, was obtained (4.2 g, 80%).

[0101] 2) Synthesis of Os(3-Me-pz-4-Me-py)2Cl2 [Chemical Formula 7] [ka] 2.0 g (4.2 mmol) of potassium hexachloroosmate(IV) and 1.4 g (8.3 mmol) of 4-methyl-2-(3-methyl-1H-pyrazol-1-yl)pyridine prepared in 1) above were placed in a 250 mL Schlenk flask and dissolved in 50 mL of ethylene glycol under an argon gas atmosphere. The mixture was then degassed with argon for 15 minutes. The reaction mixture was heated to 180°C and stirred for 1 hour. After the reaction was completed, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to 250 mL of a 1.0 M aqueous solution of sodium dithionite to obtain a reduced osmium complex precipitate. The resulting solid was filtered under vacuum, washed multiple times with water and acetonitrile, and dried in a vacuum oven to obtain the final green osmium complex (2.0 g, 78%). HRMS ( 192 Os): m / z 608.0875 ([M + ]required 608.0887)

[0102] The overall method for preparing the compound of Chemical Formula 7 is shown in Reaction Scheme 5 below. [Reaction Scheme 5] [ka]

[0103] Example 1.6. Synthesis of transition metal complex of formula 8 1) Synthesis of 4-methoxy-2-(3-methyl-1H-pyrazol-1-yl)pyridine [ka] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 2.5 g (30 mmol) of 3-methylpyrazole and 4.0 g (36 mmol) of potassium tert-butoxide, dissolved in 20 mL of anhydrous dimethyl sulfoxide under an argon atmosphere. 6.7 g (36 mmol) of 2-bromo-4-methoxypyridine was added to the reaction mixture, which was heated to 100 °C under an argon atmosphere and stirred for 18 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was concentrated under reduced pressure to remove the solvent, followed by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 3:1). Finally, a transparent solid, 4-methoxy-2-(3-methyl-1H-pyrazol-1-yl)pyridine, was obtained (3.0 g, 53%).

[0104] 2) Synthesis of Os(3-Me-p-4-MeO-py)2Cl2 [Chemical Formula 8] [ka] 2.0 g (4.2 mmol) of potassium hexachloroosmate(IV) and 1.5 g (8.3 mmol) of 4-methoxy-2-(3-methyl-1H-pyrazol-1-yl)pyridine prepared in 1) above were placed in a 250 mL Schlenk flask and dissolved in 50 mL of ethylene glycol under an argon gas atmosphere. The mixture was then degassed with argon for 15 minutes. The reaction mixture was heated to 180°C and stirred for 1 hour. After the reaction was completed, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to a 1.0 M aqueous solution of sodium dithionite (250 mL) to obtain a reduced osmium complex precipitate. The resulting solid was filtered under vacuum, washed multiple times with water and acetonitrile, and dried in a vacuum oven to obtain the final green osmium complex (1.0 g, 37%). HRMS ( 192 Os): m / z 640.0775 ([M + ]required 640.0785)

[0105] The overall method for producing the compound of Chemical Formula 8 is shown in Reaction Scheme 6 below. [Reaction Scheme 6] [ka]

[0106] Example 1.7. Synthesis of transition metal complex of formula 9 1) Synthesis of 4-methyl-2-(4-methyl-1H-pyrazol-1-yl)pyridine [ka] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 2.5 g (30 mmol) of 4-methylpyrazole and 4.0 g (36 mmol) of potassium tert-butoxide, dissolved in 20 mL of anhydrous dimethyl sulfoxide under an argon atmosphere. 6.2 g (36 mmol) of 2-bromo-4-methylpyridine was added to the reaction mixture, which was heated to 100 °C under an argon atmosphere and stirred for 18 h. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was concentrated under reduced pressure to remove the solvent, followed by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 3:1). Finally, a transparent solid, 4-methyl-2-(4-methyl-1H-pyrazol-1-yl)pyridine, was obtained (4.5 g, 86%).

[0107] 2) Synthesis of Os(4-Me-pz-4-Me-py)2Cl2 [Chemical Formula 9] [ka] 5.0 g (10 mmol) of potassium hexachloroosmate(IV) and 4.1 g (21 mmol) of 4-methyl-2-(4-methyl-1H-pyrazol-1-yl)pyridine prepared in 1) above were placed in a 250 mL Schlenk flask and dissolved in 100 mL of ethylene glycol under an argon gas atmosphere. The mixture was then degassed with argon for 15 minutes. The reaction mixture was heated to 180°C and stirred for 1 hour. After the reaction was completed, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to a 1.0 M aqueous solution of sodium dithionite (250 mL) to obtain a reduced osmium complex precipitate. The resulting solid was filtered under vacuum, washed multiple times with water and acetonitrile, and dried in a vacuum oven to obtain the final green osmium complex (5.5 g, 91%). HRMS ( 192 Os): m / z 608.0871 ([M + ]required 608.0887)

[0108] The overall method for preparing the compound of Chemical Formula 9 is shown in Reaction Scheme 7 below. [Reaction Scheme 7] [ka]

[0109] Example 1.8. Synthesis of transition metal complex of formula 10 1) Synthesis of 4-methoxy-2-(4-methyl-1H-pyrazol-1-yl)pyridine [ka] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 2.5 g (30 mmol) of 4-methylpyrazole and 4.0 g (36 mmol) of potassium tert-butoxide, dissolved in 20 mL of anhydrous dimethyl sulfoxide under an argon atmosphere. 6.7 g (36 mmol) of 2-bromo-4-methoxypyridine was added to the reaction mixture, which was heated to 100 °C under an argon atmosphere and stirred for 18 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was concentrated under reduced pressure to remove the solvent, followed by purification by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 3:1). Finally, a transparent solid, 4-methoxy-2-(4-methyl-1H-pyrazol-1-yl)pyridine, was obtained (2.8 g, 50%).

[0110] 2) Synthesis of Os(4-Me-pz4-MeO-py)2Cl2 [Chemical Formula 10] [ka] In a 250 mL Schlenk flask, 3.0 g (6.2 mmol) of potassium hexachloroosmate(IV) and 2.4 g (12 mmol) of 4-methoxy-2-(4-methyl-1H-pyrazol-1-yl)pyridine prepared in 1) above were placed and dissolved in 60 mL of ethylene glycol under an argon gas atmosphere. The mixture was then degassed with argon for 15 minutes. The reaction mixture was heated to 180°C and stirred for 1 hour. After the reaction was completed, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to a 1.0 M aqueous solution of sodium dithionite (250 mL) to obtain a reduced osmium complex precipitate. The resulting solid was filtered under vacuum, washed multiple times with water and acetonitrile, and then dried in a vacuum oven to obtain the final green osmium complex (2.3 g, 58%). HRMS ( 192 Os): m / z 640.0792 ([M + ]required 640.0785)

[0111] The overall method for preparing the compound of Chemical Formula 10 is shown in Reaction Scheme 8 below. [Reaction Scheme 8] [ka]

[0112] Example 1.9. Synthesis of transition metal complex of formula 11 1) Synthesis of (pyridin-2-yl)amidrazone [ka] 5.2 g (50 mmol) of 2-cyanopyridine and 2.7 g (55 mmol) of hydrazine hydrate were placed in a 100 mL two-neck round-bottom flask, and 4 mL of ethanol was added. The mixture was stirred at room temperature for 24 hours. After the reaction was completed, the reaction mixture was filtered under reduced pressure to remove the residual solvent and washed with benzene. The filtered solid was recrystallized from toluene to give (pyridin-2-yl)amidrazone (4.2 g, 61%).

[0113] 2) Synthesis of 2-(1,3-dimethyl-1H-1,2,4-triazol-5-yl)pyridine [ka] A 50 mL Schlenk flask was charged with 2.0 g (15 mmol) of (pyridin-2-yl)amidrazone prepared in step 1) above and 1.6 g (15 mmol) of sodium carbonate. The solvents, dimethylacetamide (15 mL) and tetrahydrofuran (5 mL), were added and stirred at 0°C. A 10 mL round-bottom flask was charged with 5 mL of anhydrous dimethylacetamide and 1.1 mL (15 mmol) of acetyl chloride. The flask was closed with a rubber septum and added dropwise to the reaction mixture using a cannula under argon. The mixture was stirred at room temperature for 5 hours. After completion of the reaction, the reaction mixture was filtered under reduced pressure to remove residual solvent and washed with ethanol and distilled water to obtain a white solid. The white solid and 20 mL of ethylene glycol were added to a 50 mL single-neck flask and heated to 190°C. The mixture was stirred for 30 minutes. After the reaction was completed, the reaction mixture was cooled to room temperature, and the ethylene glycol solvent was removed by distillation under reduced pressure to finally obtain 2-(5-R-2H-1,2,4-triazol-3-yl)pyridine (0.22 g, 9%) as a yellow solid.

[0114] 0.22 g (1.4 mmol) of 2-(3-methyl-1H-1,2,4-triazol-5-yl)pyridine was placed in a 50 mL single-neck flask and dissolved in 5 mL of anhydrous dimethylformamide under an argon atmosphere. Then, 83 mg (2.0 mmol) of sodium hydride was added. The reaction mixture was stirred at room temperature for 20 minutes, and 0.3 g (2.0 mmol) of iodomethane was added under an argon atmosphere. The mixture was then stirred at room temperature for another 24 hours. After completion of the reaction, the reaction mixture was extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was concentrated under reduced pressure and purified by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 7:3). Finally, 2-(1,3-dimethyl-1H-1,2,4-triazol-5-yl)pyridine was obtained (83 mg, 34%).

[0115] 3) Synthesis of Os(Dmtz-py)2Cl2 [Chemical Formula 11] [ka] 14 mg (28.7 μmol) of potassium hexachloroosmate(IV) and 10 mg (57 μmol) of 2-(1,3-dimethyl-1H-1,2,4-triazol-5-yl)pyridine prepared in 2) above were placed in a 5 mL Corn Vial and dissolved in 2 mL of ethylene glycol under an argon atmosphere. The mixture was then degassed with argon for 15 minutes. The reaction mixture was heated to 180°C and stirred for 1 hour. After the reaction was completed, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to 10 mL of a 1.0 M aqueous solution of sodium dithionite to obtain a reduced osmium complex precipitate. The resulting solid was filtered under vacuum, washed several times with water, and dried in a vacuum oven at 40°C to obtain the final brown osmium complex (15 mg, 86%). HRMS ( 192 Os): m / z 610.0797([M + ]required 610.0792)

[0116] The overall method for preparing the compound of Chemical Formula 11 is shown in Reaction Scheme 9 below. [Reaction Scheme 9] [ka]

[0117] Example 1.10. Synthesis of transition metal complex of formula 12 1) Synthesis of 5-methyl-3-(pyridin-2-yl)-1,2,4-oxadiazole [ka] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 7.0 g (0.1 mol) of hydroxylammonium chloride and 6.0 g (0.1 mol) of potassium hydroxide in 100 mL of methanol, heated to 100 °C, and stirred for 30 minutes. The potassium chloride formed was removed by concentration under reduced pressure, and 7.0 g (60 mmol) of pyridinecarbonitrile was added to the filtered reaction solution, heated to 100 °C, and stirred for 1 hour. After completion of the reaction, the mixture was concentrated under reduced pressure and washed with distilled water to obtain a transparent solid, hydroxypicolinimide amide (9.0 g, 65%).

[0118] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 1.0 g (7.3 mmol) of hydroxypicolinimidamide, 1.0 g (12.3 mmol) of pyridine, and 0.7 g (8.8 mmol) of acetyl chloride in 60 mL of tetrahydrofuran. The mixture was heated to 110 °C and stirred for 8 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was concentrated under reduced pressure to remove the solvent, yielding 5-methyl-3-(pyridin-2-yl)-1,2,4-oxadiazole (0.85 g, 72%) as a clear solid.

[0119] 2) Synthesis of Os(Me-oxz-py)2Cl2 [Chemical formula 12] [ka] In a 250 mL Schlenk flask, 0.6 g (1.4 mmol) of potassium hexachloroosmate(IV) and 0.5 g (2.9 mmol) of 5-methyl-3-(pyridin-2-yl)-1,2,4-oxadiazole (prepared in 1) above were dissolved in 50 mL of ethylene glycol under an argon atmosphere and degassed with argon for 15 minutes. The reaction mixture was heated to 180 °C and stirred for 20 minutes. After the reaction was complete, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to 30 mL of 1.0 M aqueous sodium dithionite solution to obtain the reduced osmium complex precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with water and acetonitrile, and then dried in a vacuum oven to obtain the final tan osmium complex (0.6 g, 70%).

[0120] The overall method for preparing the compound of Chemical Formula 12 is shown in Reaction Scheme 10 below. [Reaction Scheme 10] [ka]

[0121] Example 1.11. Synthesis of transition metal complex of formula 13 1) Synthesis of 2-(1-butyl-1H-1,2,3-triazol-4-yl)pyridine [ka] 2.0 g (14 mmol) of 1-bromobutane and 0.9 g (14 mmol) of sodium azide were placed in a 250 mL round-bottom flask, and 50 mL of anhydrous dimethylformamide was added. The mixture was stirred at room temperature for 24 hours. After the reaction was complete, the reaction mixture was extracted with water (100 mL) and diethyl ether (100 mL x 3). The organic layer was collected, dried over magnesium sulfate, and concentrated under reduced pressure to remove the solvent. The mixture was then used for the next reaction without further purification. 1-Azidobutane and 1.5 g (14 mmol) of 2-ethynylpyridine were placed in a 250 mL two-neck round-bottom flask, and tetrahydrofuran / water (40 mL / 40 mL) was added and stirred at room temperature. 0.3 g (1.4 mmol) of sodium ascorbate and 23 mg (0.14 mmol) of copper sulfate were added to the reaction mixture, which was degassed with argon for 15 minutes and then stirred at room temperature for 2 hours. After the reaction was completed, the reaction mixture was extracted with water (100 mL) and ethyl acetate (100 mL × 3). The organic layer was collected and dried over magnesium sulfate. The solution was concentrated under reduced pressure to remove the solvent, and the residue was purified by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 1:4). Finally, 2-(1-(2-methoxyethynyl)-1H-1,2,3-triazol-4-yl)pyridine (1.5 g, 52%) was obtained.

[0122] 2) Synthesis of Os(3-Bu-tz-py)2Cl2 [Chemical Formula 13] [ka] In a 100 mL Schlenk flask, 0.5 g (1.0 mmol) of potassium hexachloroosmate(IV) and 0.4 g (2.0 mmol) of 2-(1-butyl-1H-1,2,3-triazol-4-yl)pyridine (prepared in step 1) above were dissolved in 15 mL of ethylene glycol under an argon atmosphere and degassed with argon for 15 minutes. The reaction mixture was heated to 180 °C and stirred for 1 hour. After the reaction was complete, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to 200 mL of a 1.0 M aqueous solution of sodium dithionite to obtain the reduced osmium complex precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with water, and dried in a vacuum oven at 40 °C to obtain the final osmium complex (0.4 g, 56%).

[0123] The overall method for preparing the compound of Chemical Formula 13 is shown in Reaction Scheme 11 below. [Reaction Scheme 11] [ka]

[0124] Example 1.12. Synthesis of transition metal complex of formula 14 1) Synthesis of 13-bromo-2,5,8,11-tetraoxatridecane [ka] A 250 mL round-bottom flask was charged with 2.0 g (9.6 mmol) of tetraethylene glycol monomethyl ether and 3.8 g (11.5 mmol) of tetrabromomethane, dissolved in 50 mL of dichloromethane, and stirred at 0 °C in an ice bath. Then, while maintaining the temperature at 0 °C, 3.0 g (11.5 mmol) of triphenylphosphine was slowly added in small portions over 15 minutes, followed by stirring at room temperature for 2 hours. After completion of the reaction, the reaction mixture was extracted with water (100 mL) and dichloromethane (100 mL x 3). The combined organic layer was concentrated under reduced pressure and purified by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 2:1 (8% methanol)). Finally, 13-bromo-2,5,8,11-tetraoxatridecane was obtained as a yellow oil (1.4 g, 54%).

[0125] 2) Synthesis of 2-(1-(2,5,8,11-tetraoxatridecan-13-yl)-1H-1,2,3-triazol-4-yl)pyridine [ka] 1.4 g (5.2 mmol) of 13-bromo-2,5,8,11-tetraoxatridecane prepared in step 1) above and 0.34 g (5.2 mmol) of sodium azide were placed in a 250 mL round-bottom flask, and 40 mL of anhydrous dimethylformamide was added. The mixture was stirred at room temperature for 24 hours. After the reaction was complete, the reaction mixture was extracted with water (100 mL) and diethyl ether (100 mL x 3). The organic layer was collected, dried over magnesium sulfate, and concentrated under reduced pressure to remove the solvent. The mixture was then used for the next reaction without further purification. 13-Azido-2,5,8,11-tetraoxatridecane and 0.8 g (7.7 mmol) of 2-ethynylpyridine were placed in a 250 mL two-necked round-bottom flask, and tetrahydrofuran / water (40 mL / 40 mL) was added. The mixture was stirred at room temperature. To this reaction mixture, 0.15 g (0.8 mmol) of sodium ascorbate and 12 mg (0.08 mmol) of copper sulfate were added, and the mixture was degassed with argon for 15 minutes and then stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was extracted with water (100 mL) and ethyl acetate (100 mL x 3). The organic layer was collected and dried over magnesium sulfate. The solution was concentrated under reduced pressure to remove the solvent, and the residue was purified by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 1:2 (5% methanol)). Finally, 2-(1-(2,5,8,11-tetraoxatridecan-13-yl)-1H-1,2,3-triazol-4-yl)pyridine (0.86 g, 43%) was obtained.

[0126] 3) Synthesis of Os(3-tz-teg-py)2Cl2 [Chemical Formula 14] [ka] In a 50 mL Schlenk flask, 0.1 g (0.21 mmol) of potassium hexachloroosmate(IV) and 0.14 g (0.42 mmol) of 2-(1-(2,5,8,11-tetraoxatridecan-13-yl)-1H-1,2,3-triazol-4-yl)pyridine (prepared in 2) above) were dissolved in 15 mL of ethylene glycol under an argon atmosphere and degassed with argon for 15 minutes. The reaction mixture was heated to 180 °C and stirred for 1 hour. After completion of the reaction, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to 10 mL of a 1.0 M aqueous solution of sodium dithionite to obtain a reduced osmium complex precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with water, and dried in a vacuum oven at 40 °C to obtain the final dark purple osmium complex (0.1 g, 56%).

[0127] The overall method for preparing the compound of Chemical Formula 14 is shown in Reaction Scheme 12 below. [Reaction Scheme 12] [ka]

[0128] Example 1.13. Synthesis of transition metal complex of formula 15 1) Synthesis of 2-(1-(2-methoxyethynyl)-1H-1,2,3-triazol-4-yl)pyridine [ka] 2.0 g (14 mmol) of 2-bromoethyl methyl ether and 0.1 g (14 mmol) of sodium azide were placed in a 250 mL round-bottom flask, and 50 mL of anhydrous dimethylformamide was added. The mixture was stirred at room temperature for 24 hours. After the reaction was complete, the reaction mixture was extracted with water (100 mL) and diethyl ether (100 mL x 3). The organic layer was collected, dried over magnesium sulfate, and concentrated under reduced pressure to remove the solvent. The mixture was then used for the next reaction without further purification. 2-Azidoethyl methyl ether and 1.5 g (14 mmol) of 2-ethynylpyridine were placed in a 250 mL two-neck round-bottom flask, and tetrahydrofuran / water (40 mL / 40 mL) was added and stirred at room temperature. 0.28 g (1.4 mmol) of sodium ascorbate and 0.02 g (0.14 mmol) of copper sulfate were added to the reaction mixture, which was degassed with argon for 15 minutes and then stirred at room temperature for 2 hours. After the reaction was completed, the reaction mixture was extracted with water (100 mL) and ethyl acetate (100 mL × 3). The organic layer was collected and dried over magnesium sulfate. The solution was concentrated under reduced pressure to remove the solvent, and the residue was purified by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 1:4). Finally, 2-(1-(2-methoxyethynyl)-1H-1,2,3-triazol-4-yl)pyridine (1.5 g, 52%) was obtained.

[0129] 2) Synthesis of Os(3-mo-tz-py)2Cl2 [Chemical Formula 15] [ka] In a 100 mL Schlenk flask, 0.5 g (1.0 mmol) of potassium hexachloroosmate(IV) and 0.4 g (2.1 mmol) of 2-(1-(2-methoxyethynyl)-1H-1,2,3-triazol-4-yl)pyridine prepared in 1) above were placed and dissolved in 15 mL of ethylene glycol under an argon gas atmosphere. The mixture was then degassed with argon for 15 minutes. The reaction mixture was heated to 180°C and stirred for 1 hour. After the reaction was completed, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by filtration under reduced pressure. The filtrate was added dropwise to a 1.0 M aqueous solution of sodium dithionite (200 mL) to obtain a reduced osmium complex precipitate. The resulting solid was filtered under reduced pressure, washed several times with water, and dried in a vacuum oven at 40°C to obtain the final green osmium complex (0.4 g, 56%). HRMS ( 192 OS): m / z 670.09967([M + ]required 670.10)

[0130] The overall method for preparing the compound of Chemical Formula 15 is shown in Reaction Scheme 13 below. [Reaction Scheme 13] [ka]

[0131] Example 1.14. Synthesis of transition metal complex of formula 16 1) Synthesis of 2-(1H-tetrazol-5-yl)pyridine [ka] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 1.3 g (19.2 mmol) of sodium azide, 2.0 g (19.2 mmol) of 2-ethynylpyridine, and 96 mg (0.38 mmol) of copper sulfate. The resulting mixture was dissolved in 40 mL of anhydrous dimethyl sulfoxide under an argon atmosphere. The reaction mixture was degassed with argon for 15 minutes, then heated to 140 °C and stirred for 3 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was dried over magnesium sulfate. The solution was concentrated under reduced pressure to remove the solvent, ultimately yielding 2-(1H-tetrazol-5-yl)pyridine (1.1 g, 41%) as a yellow solid.

[0132] 2) Synthesis of 2-(1-methyl-1H-tetrazol-5-yl)pyridine [ka] A 100 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 1.0 g (6.8 mmol) of 2-(1H-tetrazol-5-yl)pyridine (prepared in 1) above. The solution was dissolved in anhydrous tetrahydrofuran (30 mL) under an argon atmosphere, and then 0.4 g (10 mmol) of sodium hydride was added. The reaction mixture was stirred at room temperature for 30 minutes, and then 1.5 g (10 mmol) of iodomethane was added under an argon atmosphere. The mixture was then heated to 80 °C and stirred for 3 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The organic layer was concentrated under reduced pressure to remove the solvent and purified by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 1:3). Finally, 2-(1-methyl-1H-tetrazol-5-yl)pyridine was obtained (0.4 g, 40%).

[0133] 3) Synthesis of Os(tetraz-py)2Cl2 [chemical formula 16] [ka] In a 50 mL Schlenk flask, 0.10 g (0.21 mmol) of potassium hexachloroosmate(IV) and 0.7 g (0.42 mmol) of 2-(1-methyl-1H-tetrazol-5-yl)pyridine prepared in 2) above were placed and dissolved in 5 mL of ethylene glycol under an argon atmosphere. The mixture was then degassed with argon for 15 minutes. The reaction mixture was heated to 180°C and stirred for 1 hour. After the reaction was completed, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to a 1.0 M aqueous solution of sodium dithionite (200 mL) to obtain a reduced osmium complex precipitate. The resulting solid was filtered under vacuum, washed multiple times with water, and dried in a vacuum oven at 40°C to obtain the final green osmium complex (0.1 g, 84%). HRMS ( 192 OS): m / z 584.0383([M + ]required 584.04)

[0134] The overall method for preparing the compound of Chemical Formula 16 is shown in Reaction Scheme 14 below. [Reaction Scheme 14] [ka]

[0135] Example 1.15. Synthesis of transition metal complex of formula 17 1) Synthesis of 2-(1H-1,2,4-triazol-1-yl)pyridine [ka] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 3.0 g (43 mmol) of 1H-1,2,4-triazole and 5.8 g (52 mmol) of potassium tert-butoxide, dissolved in 40 mL of anhydrous dimethyl sulfoxide under an argon atmosphere. 5.0 g (52 mmol) of 2-fluoropyridine was added to the reaction mixture, which was heated to 100 °C under an argon atmosphere and stirred for 4 h. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was concentrated under reduced pressure to remove the solvent, and then purified by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 5:1). Finally, a transparent solid, 2-(1H-1,2,4-triazol-1-yl)pyridine, was obtained (4.3 g, 57%).

[0136] 2) Synthesis of Os(1,2,4tz-py)2Cl2 [Chemical Formula 17] [ka] In a 250 mL Schlenk flask, 0.6 g (1.4 mmol) of potassium hexachloroosmate(IV) and 0.4 g (2.9 mmol) of 2-(1H-1,2,4-triazol-1-yl)pyridine (prepared in 1) above) were dissolved in 50 mL of ethylene glycol under an argon atmosphere and degassed with argon for 15 minutes. The reaction mixture was heated to 180 °C and stirred for 30 minutes. After the reaction was complete, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to 30 mL of a 1.0 M aqueous solution of sodium dithionite to obtain the reduced osmium complex precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with water and acetonitrile, and then dried in a vacuum oven to obtain the final red osmium complex (0.3 g, 62%).

[0137] The overall method for preparing the compound of Chemical Formula 17 is shown in Reaction Scheme 15 below. [Reaction Scheme 15] [ka]

[0138] Example 1.16. Synthesis of transition metal complex of formula 18 1) Synthesis of 2-(1H-1,2,3-triazol-1-yl)pyridine [ka] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 3.0 g (43 mmol) of 1H-1,2,3-triazole and 5.8 g (52 mmol) of potassium tert-butoxide, dissolved in 40 mL of anhydrous dimethyl sulfoxide under an argon atmosphere. 5.0 g (52 mmol) of 2-fluoropyridine was added to the reaction mixture, which was heated to 100 °C under an argon atmosphere and stirred for 4 h. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was concentrated under reduced pressure to remove the solvent, and then purified by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 5:1). Finally, a transparent solid, 2-(1H-1,2,3-triazol-1-yl)pyridine, was obtained (3.5 g, 56%).

[0139] 2) Synthesis of Os(1,2,3tz-py)2Cl2 [Chemical Formula 18] [ka] In a 250 mL Schlenk flask, 0.6 g (1.4 mmol) of potassium hexachloroosmate(IV) and 0.4 g (2.9 mmol) of 2-(1H-1,2,3-triazol-1-yl)pyridine (prepared in 1) above were dissolved in 50 mL of ethylene glycol under an argon atmosphere and degassed with argon for 15 minutes. The reaction mixture was heated to 180 °C and stirred for 30 minutes. After the reaction was complete, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to 30 mL of a 1.0 M aqueous solution of sodium dithionite to obtain the reduced osmium complex precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with water and acetonitrile, and then dried in a vacuum oven to obtain the green final osmium complex (0.4 g, 69%).

[0140] The overall method for preparing the compound of Chemical Formula 18 is shown in Reaction Scheme 16 below. [Reaction Scheme 16] [ka]

[0141] Example 1.17. Synthesis of transition metal complex of formula 19 1) Synthesis of 2-(1-methyl-1H-1,2,4-triazol-5-yl)pyridine [ka] 1.0 g (6.8 mmol) of 2-(1H-1,2,4-triazol-5-yl)pyridine and 0.4 g (0.01 mmol) of sodium hydride were dissolved in 50 mL of anhydrous dimethyl sulfoxide in a 100 mL two-neck round-bottom flask. 1.4 g (0.01 mmol) of methyl iodide was added dropwise to the reaction mixture using a dropping funnel, and the mixture was stirred at room temperature for 24 hours. After the reaction was complete, the reaction mixture was extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was concentrated under reduced pressure to remove the solvent, and the mixture was purified by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 5:1). Finally, a transparent solid, 2-(1-methyl-1H-1,2,4-triazol-5-yl)pyridine, was obtained (0.9 g, 86%).

[0142] 2) Synthesis of Os(4-Me-1,2,4tz-py)2Cl2 [Chemical Formula 19] [ka] In a 250 mL Schlenk flask, 0.6 g (1.4 mmol) of potassium hexachloroosmate(IV) and 0.5 g (2.9 mmol) of 2-(1-methyl-1H-1,2,4-triazol-5-yl)pyridine (prepared in 1) above) were dissolved in 50 mL of ethylene glycol under an argon atmosphere and degassed with argon for 15 minutes. The reaction mixture was heated to 180 °C and stirred for 1 hour. After the reaction was complete, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to 30 mL of 1.0 M aqueous sodium dithionite solution to obtain the reduced osmium complex precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with water and acetonitrile, and then dried in a vacuum oven to obtain the green final osmium complex (0.2 g, 25%).

[0143] The overall method for preparing the compound of Chemical Formula 19 is shown in Reaction Scheme 17 below. [Reaction Scheme 17] [ka]

[0144] Example 1.18. Synthesis of transition metal complex of formula 20 1) Synthesis of 2-(3,4-dimethyl-1H-pyrazol-1-yl)-4-methylpyridine [ka] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 2.1 g (22 mmol) of 3,4-dimethylpyrazole and 2.5 g (22 mmol) of potassium tert-butoxide, dissolved in 20 mL of anhydrous dimethyl sulfoxide under an argon atmosphere. 3.5 g (20 mmol) of 2-bromo-4-methylpyridine was added to the reaction mixture, which was heated to 100 °C under an argon atmosphere and stirred for 18 h. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was concentrated under reduced pressure to remove the solvent, followed by purification by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 3:1). Finally, a transparent solid, 4-methyl-2-(3-methyl-1H-pyrazol-1-yl)pyridine, was obtained (2.4 g, 63%).

[0145] 2) Synthesis of Os(3,4-DiMe-pz-4-Me-py)2Cl2 [Chemical Formula 20] [ka] In a 50 mL Schlenk flask, 0.5 g (1.0 mmol) of potassium hexachloroosmate(IV) and 0.4 g (2.0 mmol) of 2-(3,4-dimethyl-1H-pyrazol-1-yl)-4-methylpyridine prepared in 1) above were placed and dissolved in 15 mL of ethylene glycol under an argon gas atmosphere. The mixture was then degassed with argon for 15 minutes. The reaction mixture was heated to 180°C and stirred for 1 hour. After the reaction was completed, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to a 1.0 M aqueous solution of sodium dithionite (100 mL) to obtain a reduced osmium complex precipitate. The resulting solid was filtered under vacuum, washed multiple times with water and acetonitrile, and then dried in a vacuum oven to obtain the final green osmium complex (0.4 g, 62%). HRMS ( 192 Os): m / z 636.1205 ([M + ]required 636.1200)

[0146] The overall method for preparing the compound of formula 20 is shown in Reaction Scheme 18 below. [Reaction Scheme 18] [ka]

[0147] Example 1.19. Synthesis of transition metal complex of formula 21 1) Synthesis of 2-(3,4-dimethyl-1H-pyrazol-1-yl)-4-methoxypyridine [ka] A 50 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 0.6 g (6 mmol) of 3,4-dimethylpyrazole and 0.7 g (6 mmol) of potassium tert-butoxide, dissolved in 8 mL of anhydrous dimethyl sulfoxide under an argon atmosphere. 1.0 g (5 mmol) of 2-bromo-4-methoxypyridine was added to the reaction mixture, which was heated to 80 °C under an argon atmosphere and stirred for 6 h. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with water (50 mL) and ethyl acetate (50 mL x 3). The combined organic layer was concentrated under reduced pressure to remove the solvent, and then purified by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 3:1). Finally, a transparent solid, 4-methyl-2-(3-methyl-1H-pyrazol-1-yl)pyridine, was obtained (0.4 g, 37%).

[0148] 2) Synthesis of Os(3,4-DiMe-pz-4-MeO-py)2Cl2 [Chemical Formula 21] [ka] In a 50 mL Schlenk flask, 0.5 g (1.0 mmol) of potassium hexachloroosmate(IV) and 0.4 g (2.0 mmol) of 2-(3,4-dimethyl-1H-pyrazol-1-yl)-4-methoxypyridine prepared in 1) above were placed and dissolved in 15 mL of ethylene glycol under an argon gas atmosphere. The mixture was then degassed with argon for 15 minutes. The reaction mixture was heated to 180°C and stirred for 30 minutes. After the reaction was completed, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to a 1.0 M aqueous solution of sodium dithionite (100 mL) to obtain a reduced osmium complex precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with water and acetonitrile, and then dried in a vacuum oven to obtain the final brown osmium complex (0.4 g, 64%). HRMS ( 192 Os): m / z 668.1103 ([M + ]required 668.1098)

[0149] The overall method for preparing the compound of formula 21 is shown in Reaction Scheme 19 below. [Reaction Scheme 19] [ka]

[0150] Example 1.20. Synthesis of transition metal complex of formula 22 1) Synthesis of N,N-dimethyl-2-(4-methyl-1H-pyrazol-1-yl)pyridin-4-amine [ka] A 250 mL two-neck round-bottom flask equipped with a reflux condenser and a gas inlet was charged with 1.0 g (5.0 mmol) of 4-dimethylamino-2-bromopyridine, 1.2 g (15 mmol) of 4-methylpyrazole, 0.14 g (0.75 mmol) of copper iodide, 0.17 g (1.5 mmol) of L-proline, and 4.1 g (12.5 mmol) of cesium carbonate. The mixture was dissolved in 20 mL of anhydrous dimethylformamide under an argon atmosphere. The reaction mixture was heated to 120 °C and stirred for 20 h. After completion of the reaction, the mixture was cooled to room temperature and extracted with water (100 mL) and ethyl acetate (100 mL x 3). The combined organic layer was concentrated under reduced pressure to remove the solvent, and the mixture was purified by column chromatography using ethyl acetate and hexane as the developing solvent (hexane:ethyl acetate = 5:1). Finally, a white solid of N,N-dimethyl-2-(4-methyl-1H-pyrazol-1-yl)pyridin-4-amine was obtained (0.5 g, 50%).

[0151] 2) Synthesis of Os(4-Me-pz-4-DiAM-py)2Cl2 [Chemical Formula 22] [ka] In a 50 mL Schlenk flask, 0.6 g (1.2 mmol) of potassium hexachloroosmate(IV) and 0.5 g (2.4 mmol) of N,N-dimethyl-2-(4-methyl-1H-pyrazol-1-yl)pyridin-4-amine (prepared in 1) above were placed and dissolved in 15 mL of ethylene glycol under an argon atmosphere. The mixture was then degassed with argon for 15 minutes. The reaction mixture was heated to 180°C and stirred for 30 minutes. After the reaction was completed, the reaction mixture was cooled to room temperature, and the resulting red precipitate was removed by vacuum filtration. The filtrate was added dropwise to 100 mL of a 1.0 M aqueous solution of sodium dithionite to obtain a reduced osmium complex precipitate. The resulting solid was filtered under vacuum, washed multiple times with water and acetonitrile, and then dried in a vacuum oven to obtain the final red-black osmium complex (0.5 g, 75%). HRMS ( 192 Os): m / z 666.1421 ([M + ]required 666.1418)

[0152] The overall method for preparing the compound of formula 22 is shown in Reaction Scheme 20 below. [Reaction Scheme 20] [ka]

[0153] Example 1.21. Synthesis of transition metal complex of formula 23 1) Synthesis of Ru(DMSO)Cl [ka] Add RuCl3 to a 50 mL Schlenk flask. *0.9 g (4.3 mmol) of HO and 5 mL of anhydrous dimethyl sulfoxide were added and degassed under an argon atmosphere for 10 minutes. The dark red suspension was heated to 170 °C and stirred for 30 minutes. This temperature was maintained until the reaction solution turned dark yellow, after which the temperature was lowered to room temperature to terminate the reaction. 4 mL of acetone was added to the reaction solution, which was then cooled to 0 °C and left for 4-5 hours. The resulting solid was filtered under reduced pressure and washed with cold acetone. Finally, a yellow solid, Ru(DMSO)4Cl2, was obtained, which was used in the next reaction without further purification (1.5 g, 75%).

[0154] 2) Synthesis of Ru(pzpy)2Cl2 [Chemical Formula 23] [ka] A 50 mL Schlenk flask was charged with 0.4 g (0.86 mmol) of Ru(DMSO)4Cl2, 0.25 g (1.7 mmol) of 2-(1H-pyrazol-1-yl)pyridine prepared in the previous example, 1.8 g (43.0 mmol) of lithium chloride, and 15 mL of anhydrous dimethylformamide. The flask was degassed under an argon atmosphere for 10 minutes. The reaction vessel was wrapped in aluminum foil to protect from light and then stirred at 150 °C for 4 hours. After the reaction was complete, 50 mL of acetone was added to the dark purple reaction solution and cooled at 0 °C for 24 hours. The dark purple solid was filtered under reduced pressure, and the filtrate was washed with acetone to remove lithium chloride and by-products until the filtrate was clear. The remaining dark purple solid was dried in a vacuum oven to obtain the final ruthenium complex. The mixture was then left for 4-5 hours. The resulting solid was filtered under reduced pressure and washed with cold acetone. Finally, a yellow solid, Ru(DMSO)Cl, was obtained (0.25 g, 55%). HRMS: m / z 461.9710 ([M + ]required 461.9690)

[0155] The overall method for preparing the compound of formula 23 is shown in Reaction Scheme 21 below. [ka]

[0156] Example 1.22. Synthesis of transition metal complex of formula 24 1) Synthesis of Ru(4-Me-pz4-Me-py)2Cl2 [Chemical Formula 24] [ka] A 50 mL Schlenk flask was charged with 0.4 g (0.86 mmol) of Ru(DMSO)4Cl2, 0.3 g (1.7 mmol) of 4-methyl-2-(4-methyl-1H-pyrazol-1-yl)pyridine (prepared in the previous example), 1.8 g (43.0 mmol) of lithium chloride, and 15 mL of anhydrous dimethylformamide. The flask was degassed under an argon atmosphere for 10 minutes. The reaction vessel was wrapped in aluminum foil to protect from light and then stirred at 150°C for 4 hours. After the reaction was complete, 50 mL of acetone was added to the dark purple reaction solution and cooled at 0°C for 24 hours. The dark purple solid was filtered under reduced pressure and washed with acetone until the filtrate was clear to remove lithium chloride and by-products. The remaining black solid was dried in a vacuum oven to obtain the final ruthenium complex (0.1 g, 23%). HRMS: m / z 518.0321 ([M + ]required 518.0316)

[0157] The overall method for preparing the compound of formula 24 is shown in Reaction Scheme 22 below. [ka]

[0158] Example 1.23. Synthesis of transition metal complex of formula 25 1) Synthesis of Fe(pzpy)2Cl2 [Chemical Formula 25] [ka] A 100 mL Schlenk flask was charged with 1.6 g (10.0 mmol) of FeCl3, 1.5 g (10.0 mmol) of 2-(1H-pyrazol-1-yl)pyridine prepared in the previous example, 3.3 g (20.0 mmol) of terephthalic acid, 0.8 g (20 mmol) of sodium hydroxide, and 30 mL of absolute ethanol. The mixture was degassed under an argon atmosphere for 10 minutes. The reaction solution was stirred at 50°C for 96 hours. After the reaction was completed, the temperature was lowered to room temperature, and the resulting reddish-black solid was filtered under reduced pressure and washed with acetone. The reddish-black solid was dried in a vacuum oven to obtain the final iron complex (1.0 g, 24%). HRMS: m / z 415.9971 ([M + ]required 415.9995)

[0159] Example 2: Synthesis of an oxidation-reduction polymer containing a transition metal complex according to the present invention Example 2.1. Synthesis of the oxidation-reduction polymer of formula 28 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.12 g (0.22 mmol) of Os(pzpy)2Cl2 [Formula 3] prepared in Example 1.1 above and completely dissolved in 10 mL of ethanol under an argon gas atmosphere. 0.1 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 20 mL of ethanol was added to the reaction mixture, heated to 100 °C, and stirred for 2 days. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.2 g of a dark green oxidation-reduction polymer of Formula 25 (PVI-Os(pzpy)2Cl) (0.20 g, 91%).

[0160] Example 2.2. Synthesis of the oxidation-reduction polymer of formula 29 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.13 g (0.22 mmol) of Os(pz-2-Me-py)2Cl2 [Formula 4] prepared in Example 1.2 and completely dissolved in 10 mL of ethanol under an argon gas atmosphere. 0.1 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 20 mL of ethanol was added to the reaction mixture, heated to 100 °C, and stirred for 2 days. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.2 g of a dark green oxidation-reduction polymer of Formula 26 (PVI-Os(pz-2-Me-py)2Cl) (0.20 g, 87%).

[0161] Example 2.3. Synthesis of the oxidation-reduction polymer of formula 30 [ka] In a 100 mL Schlenk flask, 0.13 g (0.22 mmol) of Os(pz-4-MeO-py)2Cl2 [Formula 5] prepared in Example 1.3 was placed and completely dissolved in 10 mL of ethanol under an argon gas atmosphere. 0.1 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 20 mL of ethanol was added to the reaction mixture, which was then heated to 120 °C and stirred for 24 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.2 g of a dark green oxidation-reduction polymer of Formula 27 (PVI-Os(pz-4-MeO-py)2Cl) (0.21 g, 89%).

[0162] Example 2.4. Synthesis of the oxidation-reduction polymer of formula 31 [ka] In a 100 mL Schlenk flask, 0.31 g (0.53 mmol) of Os(pz-4-Me-py)2Cl2 [Formula 6] prepared in Example 1.4 was placed and completely dissolved in 20 mL of ethanol under an argon gas atmosphere. 0.2 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 30 mL of ethanol was added to the reaction mixture, which was then heated to 120 °C and stirred for 36 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.4 g of a dark green oxidation-reduction polymer of Formula 28 (PVI-Os(pz-4-Me-py)2Cl) (0.4 g, 78%).

[0163] Example 2.5. Synthesis of the oxidation-reduction polymer of formula 32 [ka] In a 100 mL Schlenk flask, 0.27 g (0.44 mmol) of Os(3-Me-pz-4-Me-py)2Cl2 [Formula 7] prepared in Example 1.5 was placed and completely dissolved in 20 mL of ethanol under an argon gas atmosphere. 0.2 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 30 mL of ethanol was added to the reaction mixture, heated to 120 °C, and stirred for 24 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, finally yielding 0.4 g of a dark green oxidation-reduction polymer of Formula 29 (PVI-Os(3-Me-pz-4-Me-py)2Cl) (0.41 g, 87%).

[0164] Example 2.6. Synthesis of the oxidation-reduction polymer of formula 33 [ka] In a 100 mL Schlenk flask, 0.14 g (0.22 mmol) of Os(3-Me-pz-4-MeO-py)2Cl2 [Formula 8] prepared in Example 1.6 was placed and completely dissolved in 10 mL of ethanol under an argon gas atmosphere. 0.1 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 20 mL of ethanol was added to the reaction mixture, which was then heated to 120 °C and stirred for 24 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.22 g of a dark green oxidation-reduction polymer of Formula 30 (PVI-Os(3-Me-pz-4-MeO-py)2Cl) (0.22 g, 92%).

[0165] Example 2.7. Synthesis of the oxidation-reduction polymer of formula 34 [ka] In a 250 mL Schlenk flask, 0.8 g (1.32 mmol) of Os(4-Me-pz-4-Me-py)2Cl2 [Formula 9] prepared in Example 1.7 was placed and completely dissolved in 50 mL of ethanol under an argon gas atmosphere. 0.5 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 50 mL of ethanol was added to the reaction mixture, which was then heated to 120 °C and stirred for 24 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 1.21 g of a dark green oxidation-reduction polymer of Formula 31 (PVI-Os(4-Me-pz-4-Me-py)2Cl) (1.21 g, 93%).

[0166] Example 2.8. Synthesis of the oxidation-reduction polymer of formula 35 [ka] In a 100 mL Schlenk flask, 0.14 g (0.22 mmol) of Os(4-Me-pz-4-MeO-py)2Cl2 [Formula 10] prepared in Example 1.8 was placed and completely dissolved in 10 mL of ethanol under an argon gas atmosphere. 0.1 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 20 mL of ethanol was added to the reaction mixture, heated to 120 °C, and stirred for 24 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.20 g of a dark green oxidation-reduction polymer of Formula 32 (PVI-Os(4-Me-pz-4-MeO-py)2Cl) (0.20 g, 83%).

[0167] Example 2.9. Synthesis of the oxidation-reduction polymer of formula 36 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.13 g (0.22 mmol) of Os(Dmtz-py)2Cl2 [formula 11] prepared in Example 1.9 and completely dissolved in 10 mL of ethanol under an argon gas atmosphere. 0.1 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 20 mL of ethanol was added to the reaction mixture, heated to 100 °C, and stirred for 2 days. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.15 g of a brown oxidation-reduction polymer of formula 33 (PVI-Os(Dmtz-py)2Cl) (0.15 g, 65%).

[0168] Example 2.10. Synthesis of the oxidation-reduction polymer of formula 37 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.12 g (0.18 mmol) of Os(3-Bu-tz-py)2Cl2 [formula 13] prepared in Example 1.11 above and completely dissolved in ethanol under an argon gas atmosphere. 85 mg of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 20 mL of ethanol was added to the reaction mixture, which was then heated to 100 °C and stirred for 2 days. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.2 g of red PVI-Os polymer (PVI-Os(3-Bu-tz-py)2Cl) of formula 34 (0.2 g, 95%).

[0169] Example 2.11. Synthesis of the oxidation-reduction polymer of formula 38 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.15 g (0.22 mmol) of Os(3-motz-py)2Cl2 [formula 15] prepared in Example 1.13 and completely dissolved in 10 mL of ethanol under an argon gas atmosphere. 0.1 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 20 mL of ethanol was added to the reaction mixture, heated to 100 °C, and stirred for 2 days. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.12 g of a brown oxidation-reduction polymer of formula 35 (PVI-Os(3-motz-py)2Cl) (0.12 g, 48%).

[0170] Example 2.12. Synthesis of the oxidation-reduction polymer of formula 39 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.12 g (0.22 mmol) of Os(1,2,3-tz-py)2Cl2 [formula 18] prepared in Example 1.16 and completely dissolved in 10 mL of ethanol under an argon gas atmosphere. 0.1 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 20 mL of ethanol was added to the reaction mixture, which was then heated to 110 °C and stirred for 2 days. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.22 g of red oxidation-reduction polymer of formula 36 (PVI-Os(1,2,3-tz-py)2Cl) (0.21 g, 96%).

[0171] Example 2.13. Synthesis of the oxidation-reduction polymer of formula 40 [ka] In a 100 mL Schlenk flask, 0.17 g (0.27 mmol) of Os(3,4-DiMe-pz-4-Me-py)2Cl2 [formula 20] prepared in Example 1.18 was placed and completely dissolved in 15 mL of ethanol under an argon gas atmosphere. 0.1 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 20 mL of ethanol was added to the reaction mixture, which was then heated to 120 °C and stirred for 24 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.22 g of a dark green oxidation-reduction polymer of formula 37 (PVI-Os(3,4-DiMe-pz-4-Me-py)2Cl) (0.22 g, 81%).

[0172] Example 2.14. Synthesis of the oxidation-reduction polymer of formula 41 [ka] In a 100 mL Schlenk flask, 0.29 g (0.43 mmol) of Os(3,4-Dime-pz-4-MeO-py)2Cl2 [Formula 21] prepared in Example 1.19 was placed and completely dissolved in 20 mL of ethanol under an argon gas atmosphere. 0.2 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 30 mL of ethanol was added to the reaction mixture, which was then heated to 120 °C and stirred for 48 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.40 g of a brown oxidation-reduction polymer of Formula 38 (PVI-Os(3,4-Dime-pz-4-MeO-py)2Cl) (0.40 g, 81%).

[0173] Example 2.15. Synthesis of the oxidation-reduction polymer of formula 42 [ka] In a 100 mL Schlenk flask, 0.18 g (0.27 mmol) of Os(4-Me-pz-4-DiAM-py)2Cl2 [formula 22] prepared in Example 1.20 was placed and completely dissolved in 20 mL of ethanol under an argon gas atmosphere. 0.1 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 20 mL of ethanol was added to the reaction mixture, which was then heated to 120 °C and stirred for 18 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.24 g of a brown oxidation-reduction polymer of formula 39 (PVI-Os(4-Me-pz-4-DiAM-py)2Cl) (0.24 g, 86%).

[0174] Example 2.16. Synthesis of the oxidation-reduction polymer of formula 43 [ka] In a 250 mL Schlenk flask, 0.5 g (0.76 mmol) of Os(4-Me-pz-4-DiAM-py)2Cl2 [formula 22] prepared in Example 1.20 was placed and completely dissolved in 60 mL of ethanol under an argon gas atmosphere. 0.32 g of polyvinylpyridine (Mn = 160,000 g / mol) completely dissolved in 30 mL of ethanol was added to the reaction mixture, which was then heated to 120 °C and stirred for 24 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.70 g of a brown oxidation-reduction polymer of formula 40 (PVP-Os(4-Me-pz-4-DiAM-py)2Cl) (0.70 g, 85%).

[0175] Example 2.17. Synthesis of the oxidation-reduction polymer of formula 44 [ka] In a 100 mL Schlenk flask, 0.14 g (0.27 mmol) of Ru(4-Me-pz4-Me-py)2Cl2 [Formula 24] prepared in Example 1.22 was placed and completely dissolved in 30 mL of ethanol under an argon gas atmosphere. 0.1 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 20 mL of ethanol was added to the reaction mixture, which was then heated to 100 °C and stirred for 12 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40 °C for 24 hours, yielding 0.24 g of a dark green oxidation-reduction polymer of Formula 44 (PVI-Ru(4-Me-pz4-Me-py)2Cl) (0.2 g, 83%).

[0176] Example 2.18. Synthesis of the oxidation-reduction polymer of formula 45 [ka] In a 100 mL Schlenk flask, 0.11 g (0.27 mmol) of Fe(pzpy)2Cl2 [formula 25] prepared in Example 1.23 was placed and completely dissolved in 25 mL of ethanol under an argon gas atmosphere. 0.1 g of polyvinylimidazole (Mn = 10,000 g / mol) completely dissolved in 20 mL of ethanol was added to the reaction mixture, which was then heated to 80°C and stirred for 24 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure, washed multiple times with diethyl ether, and dried in a vacuum oven at 40°C for 24 hours, yielding 0.1 g of reddish-brown oxidation-reduction polymer of formula 45 (PVI-Fe(pzpy)2Cl) (0.1 g, 47%).

[0177] Example 3. Synthesis of a redox polymer containing a transition metal complex and a crosslinkable functional group according to the present invention Example 3.1. Synthesis of the oxidation-reduction polymer of formula 48 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.2 g of the polymer of formula 28 prepared in Example 2.1 above and completely dissolved in methanol under an argon gas atmosphere. 20 mg (0.1 mmol) of 2-bromoethylamine was added to the reaction mixture, which was heated to 80°C and stirred for 24 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure and washed multiple times with diethyl ether. To exchange the Br ions contained in the reaction mixture for Cl ions, the filtered solid and 50 mL of water were placed in a 200 mL beaker and dissolved completely. 20 mL of ion exchange resin (AG1x4) was added and stirred for 24 hours. The reaction mixture was filtered under reduced pressure to remove the resin, and the filtered aqueous solution was lyophilized to remove the water, yielding 0.2 g of a green polymer of formula 48 (0.2 g, 90%).

[0178] Example 3.2. Synthesis of the oxidation-reduction polymer of formula 49 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.2 g of the polymer of formula 30 prepared in Example 2.3 above and completely dissolved in methanol under an argon gas atmosphere. 20 mg (0.1 mmol) of 2-bromoethylamine was added to the reaction mixture, which was heated to 80°C and stirred for 24 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure and washed multiple times with diethyl ether. To exchange the Br ions contained in the reaction mixture for Cl ions, the filtered solid and 50 mL of water were placed in a 200 mL beaker and dissolved completely. 20 mL of ion exchange resin (AG1x4) was added and stirred for 24 hours. The reaction mixture was filtered under reduced pressure to remove the resin, and the filtered aqueous solution was lyophilized to remove the water, yielding 0.2 g of a green polymer of formula 49 (0.2 g, 90%).

[0179] Example 3.3. Synthesis of the oxidation-reduction polymer of formula 50 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.4 g of the polymer of formula 31 prepared in Example 2.4 above and completely dissolved in methanol under an argon gas atmosphere. 50 mg (0.25 mmol) of 2-bromoethylamine was added to the reaction mixture, which was heated to 80°C and stirred for 24 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure and washed multiple times with diethyl ether. To exchange the Br ions contained in the reaction mixture for Cl ions, the filtered solid and 50 mL of water were placed in a 200 mL beaker and dissolved completely. 20 mL of ion exchange resin (AG1x4) was added and stirred for 24 hours. The reaction mixture was filtered under reduced pressure to remove the resin, and the filtered aqueous solution was lyophilized to remove the water, yielding 0.41 g of a green polymer of formula 50 (0.41 g, 91%).

[0180] Example 3.4. Synthesis of the oxidation-reduction polymer of formula 51 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.4 g of the polymer of formula 34 prepared in Example 2.7 above and completely dissolved in methanol under an argon gas atmosphere. 50 mg (0.25 mmol) of 2-bromoethylamine was added to the reaction mixture, which was heated to 80°C and stirred for 24 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure and washed multiple times with diethyl ether. To exchange the Br ions contained in the reaction mixture for Cl ions, the filtered solid and 50 mL of water were placed in a 200 mL beaker and dissolved completely. 20 mL of ion exchange resin (AG1x4) was added and stirred for 24 hours. The reaction mixture was filtered under reduced pressure to remove the resin, and the filtered aqueous solution was lyophilized to remove the water, yielding 0.4 g of a green polymer of formula 51 (0.4 g, 90%).

[0181] Example 3.5. Synthesis of the oxidation-reduction polymer of formula 52 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.2 g of the polymer of formula 35 prepared in Example 2.8 above and completely dissolved in methanol under an argon gas atmosphere. 20 mg (0.1 mmol) of 2-bromoethylamine was added to the reaction mixture, which was heated to 80°C and stirred for 24 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure and washed multiple times with diethyl ether. To exchange the Br ions contained in the reaction mixture for Cl ions, the filtered solid and 50 mL of water were placed in a 200 mL beaker and dissolved completely. 20 mL of ion exchange resin (AG1x4) was added and stirred for 24 hours. The reaction mixture was filtered under reduced pressure to remove the resin, and the filtered aqueous solution was lyophilized to remove the water, yielding 0.2 g of a green polymer of formula 52 (0.2 g, 90%).

[0182] Example 3.6. Synthesis of the oxidation-reduction polymer of formula 53 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.2 g of the polymer of formula 40 prepared in Example 2.13 above and completely dissolved in methanol under an argon gas atmosphere. 20 mg (0.1 mmol) of 2-bromoethylamine was added to the reaction mixture, which was heated to 80°C and stirred for 24 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure and washed multiple times with diethyl ether. To exchange the Br ions contained in the reaction mixture for Cl ions, the filtered solid and 50 mL of water were placed in a 200 mL beaker and dissolved completely. 20 mL of ion exchange resin (AG1x4) was added and stirred for 24 hours. The reaction mixture was filtered under reduced pressure to remove the resin, and the filtered aqueous solution was lyophilized to remove the water, yielding 0.2 g of a green polymer of formula 53 (0.2 g, 90%).

[0183] Example 3.7. Synthesis of the oxidation-reduction polymer of formula 54 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.2 g of the polymer of formula 41 prepared in Example 2.14 above and completely dissolved in methanol under an argon gas atmosphere. 20 mg (0.1 mmol) of 2-bromoethylamine was added to the reaction mixture, which was heated to 80°C and stirred for 24 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure and washed multiple times with diethyl ether. To exchange the Br ions contained in the reaction mixture for Cl ions, the filtered solid and 50 mL of water were placed in a 200 mL beaker and dissolved completely. 20 mL of ion exchange resin (AG1x4) was added and stirred for 24 hours. The reaction mixture was filtered under reduced pressure to remove the resin, and the filtered aqueous solution was lyophilized to remove the water, yielding 0.2 g of a green polymer of formula 54 (0.2 g, 90%).

[0184] Example 3.8. Synthesis of the oxidation-reduction polymer of formula 57 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.2 g of the polymer of formula 42 prepared in Example 2.15 above and completely dissolved in methanol under an argon gas atmosphere. 20 mg (0.1 mmol) of 2-bromoethylamine was added to the reaction mixture, which was heated to 80°C and stirred for 24 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure and washed multiple times with diethyl ether. To exchange the Br ions contained in the reaction mixture for Cl ions, the filtered solid and 50 mL of water were placed in a 200 mL beaker and dissolved completely. 20 mL of ion exchange resin (AG1x4) was added and stirred for 24 hours. The reaction mixture was filtered under reduced pressure to remove the resin, and the filtered aqueous solution was lyophilized to remove the water, yielding 0.2 g of a red polymer of formula 57 (0.2 g, 90%).

[0185] Example 3.9. Synthesis of the oxidation-reduction polymer of formula 58 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.4 g of the polymer of formula 43 prepared in Example 2.16 and completely dissolved in methanol under an argon gas atmosphere. 30 mg (0.15 mmol) of 2-bromoethylamine was added to the reaction mixture, which was heated to 80°C and stirred for 24 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure and washed multiple times with diethyl ether. To exchange the Br ions contained in the reaction mixture for Cl ions, the filtered solid and 50 mL of water were placed in a 200 mL beaker and dissolved completely. 20 mL of ion exchange resin (AG1x4) was added and stirred for 24 hours. The reaction mixture was filtered under reduced pressure to remove the resin, and the filtered aqueous solution was lyophilized to remove the water, yielding 0.41 g of a red polymer of formula 58 (0.41 g, 95%).

[0186] Example 3.10. Synthesis of the oxidation-reduction polymer of formula 59 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.4 g of the polymer of formula 34 prepared in Example 2.7 above and completely dissolved in methanol under an argon gas atmosphere. 24 mg (0.1 mmol) of diethylene glycol-2-bromoethyl methyl ether was added to the reaction mixture, which was heated to 80°C and stirred for 24 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure and washed multiple times with diethyl ether. To exchange the Br ions contained in the reaction mixture for Cl ions, the filtered solid and 50 mL of water were placed in a 200 mL beaker and dissolved completely. 20 mL of ion exchange resin (AG1x4) was added and stirred for 24 hours. The reaction mixture was filtered under reduced pressure to remove the resin, and the filtered aqueous solution was lyophilized to remove the water, yielding 0.4 g of a green polymer of formula 59 (0.4 g, 94%).

[0187] Example 3.11. Synthesis of the oxidation-reduction polymer of formula 60 [ka] A 100 mL three-neck round-bottom flask equipped with a reflux condenser, gas inlet, and thermometer was charged with 0.4 g of the polymer of formula 44 prepared in Example 2.17 and completely dissolved in methanol under an argon gas atmosphere. 30 mg (0.15 mmol) of 2-bromoethylamine was added to the reaction mixture, which was heated to 80°C and stirred for 24 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and added dropwise to diethyl ether to obtain a polymer precipitate. The resulting solid was filtered under reduced pressure and washed multiple times with diethyl ether. To exchange the Br ions contained in the reaction mixture for Cl ions, the filtered solid and 50 mL of water were placed in a 200 mL beaker and dissolved completely. 20 mL of ion exchange resin (AG1x4) was added and stirred for 24 hours. The reaction mixture was filtered under reduced pressure to remove the resin, and the filtered aqueous solution was lyophilized to remove the water, yielding 0.35 g of a red polymer of formula 60 (0.35 g, 81%).

[0188] Experimental Example 1: Confirmation of the electrochemical properties of the transition metal complex and redox polymer of the present invention using the cyclic voltammetry method In order to confirm the performance of the transition metal complexes having bidentate ligands including pyrazole, triazole, tetrazole, oxadiazole or thiadiazole according to the present invention and the oxidation-reduction polymers containing the same as electron transfer mediators, the electrochemical characteristics were measured using a cyclic voltammetry method according to the following experimental method.

[0189] Experimental Method 1. 20 mg of each of the compounds (transition metal complexes) of the chemical formulae 3, 4, 9, 11, 14, 15, 16, 17, 18, 20, 22, 23, 24, and 25 according to the present invention was dissolved in 2 mL of a 0.1 M solution of tetrabutylammonium perchlorate in dimethyl sulfoxide.

[0190] 20 mg of compounds of formulas 28, 34, 40, and 42 (oxidation-reduction polymers) according to the present invention and compounds of formulas 48, 51, 57, and 59 (oxidation-reduction polymers further containing crosslinkable functional groups) were each dissolved in 5 mL of deionized water and 0.1 M sodium chloride solution. As a comparison group, 20 mg of the compound of formula 61 below was dissolved in 5 mL of deionized water and 0.1 M sodium chloride solution.

[0191] [Chemical formula 61] [ka] 2. The solution was degassed with argon for 5 to 10 minutes to remove oxygen from the solution.

[0192] 3. A working electrode, a reference electrode, and a counter electrode were connected to the oxygen-deaerated solution, and the change in electrical signal corresponding to the change in voltage was measured in an argon gas atmosphere.

[0193] 4. The results of this experiment are shown in Tables 1 to 3, Figures 1a to 1o, Figures 2 and 3 below.

[0194] The experimental results for each compound are shown in the corresponding figures below: Chemical formula 3 (Figure 1a), chemical formula 4 (Figure 1b), chemical formula 9 (Figure 1c), chemical formula 11 (Figure 1d), chemical formula 14 (Figure 1e), chemical formula 15 (Figure 1f), chemical formula 16 (Figure 1g), chemical formula 17 (Figure 1h), chemical formula 18 (Figure 1i), chemical formula 20 (Figure 1j), chemical formula 22 (Figure 1k), chemical formula 23 (Figure 1l), chemical formula 24 (Figure 1m), chemical formula 25 (Figure 1n), chemical formulas 3, 4, 11, 14, 15, 16 (Figure 1o), Chemical formula 3 (comparison group), 28, 34, 40, 42 (Figure 2), Chemical formulas 28 (comparison group), 48, 51, 57, 59, and 61 (comparison group) (Figure 3).

[0195] Experimental materials / conditions Working electrode: glassy carbon electrode (diameter: 3.0 mm) Reference electrode:Ag / AgCl electrode Counter electrode: Platinum rod Test parameters -Equipment: EmStat (PalmSens Co.) -Technique: cyclic voltammetry - Potential range: -1.0~1.0V -Scan rate: 10mV / s

[0196] [Table 1]

[0197] [Table 2]

[0198] [Table 3] As shown in Table 1 and FIGS. 1a to 1o, it was confirmed that the transition metal complexes according to the present invention have various potential values ​​depending on the type of ligand.

[0199] As shown in Table 2 above and FIG. 2, it was confirmed that when the transition metal complex according to the present invention was synthesized using an oxidation-reduction polymer, the potential value inherent to the complex changed.

[0200] Furthermore, as shown in Table 3 and Figure 3, it was confirmed that the introduction of a crosslinkable functional group into the oxidation-reduction polymer of the present invention does not affect the potential value of the oxidation-reduction polymer. Furthermore, it was confirmed that the compounds of Chemical Formulas 48, 51, 57, and 59 exhibit lower potential values ​​than the conventionally known compound of Chemical Formula 61, and can function as highly efficient oxidation-reduction mediators.

[0201] Experimental Example 2: Preparation of an electrochemical sensor for continuous blood glucose monitoring containing the oxidation-reduction polymer according to the present invention In order to prepare an electrochemical sensor (electrochemical biosensor) containing an electron transfer mediator of the oxidation-reduction polymer according to the present invention, the sensor was prepared by the following method.

[0202] Experimental Method 1. The compounds of Formulas 48, 51, 53, and 59 according to the present invention and the compound of Formula 61 as a comparative example were dissolved in aqueous solutions containing an oxidoreductase (glucose dehydrogenase), carbon nanotubes (CNTs), a nonionic surfactant (Triton-X), and a cross-linking agent (polyethylene glycol diglycidylether), and the solutions were prepared using stirring and ultrasonic dispersion methods.

[0203] 2. To fabricate a continuous blood glucose electrochemical sensor, each solution was dispensed onto a carbon paste-printed electrode and then cured by crosslinking at room temperature for 24 hours. After curing, the sensor was washed with distilled water.

[0204] 3. Cyclic voltammetry was used to compare the electron transfer performance of the electrode fabricated above with that of an electrode containing formula 61.

[0205] 4. The results of this experiment are shown in Table 4 below and Figure 4.

[0206] Experimental materials / conditions Working electrode: The electrode fabricated above Reference electrode:Ag / AgCl electrode Counter electrode: Platinum wire Electrolyte: Phosphate buffer with NaCl solution Test parameters -Equipment: EmStat (PalmSens Co.) -Technique: cyclic voltammetry - Potential range: -0.3~0.4V -Scan rate: 10mV / s

[0207] [Table 4] As shown in Table 4 and FIG. 4, the potential E of the electrode to which the oxidation-reduction polymer of the present invention is applied 0 has a lower potential than the comparison electrode. This is the same result as in Experimental Example 1, and this experiment confirmed that the electrochemical sensor for continuous blood glucose monitoring can be used stably even at an operating voltage lower than that of the comparison electrode.

[0208] Experimental Example 3: Comparison of sensitivity of electrochemical sensors for continuous blood glucose monitoring to changes in glucose concentration 1. The electrodes prepared in Experimental Example 2 using the compounds of Formulas 51 and 53 according to the present invention and the compound of Formula 61 as a comparative group were compared by chronoamperometry in 0 to 100 mM glucose solutions.

[0209] 2. When performing the time-zone amperometry, the voltage applied was 0.15 V for the electrodes using the compounds of formulas 51 and 53, and 0.25 V for the electrode using the compound of formula 61 as a comparison group.

[0210] 3. The glucose concentrations were 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 50, and 100 mM. Each experiment lasted 50 minutes.

[0211] 4. The results of this experiment are shown in Figures 5 and 6, respectively.

[0212] Experimental materials / conditions Working electrode: The electrode fabricated above Reference electrode:Ag / AgCl electrode Counter electrode: Platinum wire Electrolyte: Phosphate buffer with NaCl solution Test parameters -Equipment: EmStat (PalmSens Co.) -Technique: chronoamperometry -Potential range: 0.15V, 0.25V

[0213] As shown in Figures 5 and 6, the electrodes to which the oxidation-reduction polymers of the present invention were applied showed linearity to glucose concentrations of 10 mM or less, and showed similar sensitivity to the control electrodes despite the application of lower voltages than the control electrodes. In particular, the electrodes to which the compounds of formulas 51 and 53 were applied showed larger currents at a concentration of 100 mM than the control electrodes, indicating the maximum enzyme activity V max is considered to be 1.2 to 2 times larger.

[0214] Experimental Example 4: Comparison of glucose sensitivity by increasing voltage of electrochemical sensors for continuous blood glucose monitoring 1. The electrodes prepared in Experimental Example 2 using the compounds of Formulas 51 and 53 according to the present invention and the compound of Formula 61 as a comparative group were compared by performing a multi-potential step method in a 400 mM glucose solution.

[0215] 2. When performing the multi-potential step method, the voltage ranged from -0.2V to 0.35V, with 0.05V intervals between the voltages, and the current was observed while maintaining the voltage for 300 seconds.

[0216] 3. The results of this experiment are shown in Figure 7.

[0217] Experimental materials / conditions Working electrode: The electrode fabricated above Reference electrode:Ag / AgCl electrode Counter electrode: Platinum wire Electrolyte: Phosphate buffer with NaCl solution Test parameters -Equipment: EmStat (PalmSens Co.) -Technique:multi-potential step - Potential range: -0.2~0.35V

[0218] As shown in FIG. 7, it was confirmed that all of the electrodes using the oxidation-reduction polymer according to the present invention exhibited a glucose sensitivity at a lower voltage than the control electrodes.

Claims

1. A transition metal complex or a salt compound thereof, which is a compound represented by the following chemical formula 1: [Chemical formula 1] (L) ａ (X １ ) ｂ M c ・d(X ２ ) In the above formula, M is Os; L is a bidentate ligand comprising (i) pyridine and (ii) one structure selected from the group consisting of pyrazole, triazole, tetrazole, oxadiazole, and thiadiazole, and the structures (i) and (ii) are directly linked; The pyridine is unsubstituted or １－４ Alkyl group, C １－４ Alkoxy group, -(CH2)-O-C １－４ Alkyl group, -(CH2CH2)-O-C １－４ Alkyl groups, and C １－４ a pyridine substituted with 1 to 4 alkylamino groups; Each of the members selected from the group consisting of pyrazole, triazole, tetrazole, and oxadiazole is independently unsubstituted or C １－４ Alkyl group, C １－４ Alkoxy group, -(CH2)-O-C １－４ Alkyl group, -(CH2CH2)-O-C １－４ Alkyl groups, 【Chemistry 1】 , and C １－４ substituted with 1 to 3 substituents selected from the group consisting of alkylamino groups; R' ４ is hydrogen or substituted or unsubstituted C １－４ is alkyl, n' is an integer selected from 1 to 4; a is 2; The substituted C １－４ Alkyl group, C １－４ Alkoxy group, -(CH2)-O-C １－４ Alkyl group, -(CH2CH2)-O-C １－４ Alkyl group, or C １－４ The alkylamino group is one in which a hydrogen atom is substituted by a halogen atom such as 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, or a phosphoric acid group or a salt thereof; X １ is one halogen atom selected from the group consisting of F, Cl, Br and I, b is 2; c is +2 and d is 0, or c is +3 and d is 1; X ２ are F, Cl, Br, I and PF ６ is one counter ion selected from the group consisting of:

2. The transition metal complex according to claim 1 , which is a compound represented by the following formula 2: [Chemical formula 2] 【Chemistry 2】 In the above formula, R １ , R ２ , R ３ and R ４ are each independently hydrogen, C １－４ Alkyl group, C １－４ Alkoxy group, -(CH2)-O-C １－４ Alkyl group, -(CH2CH2)-O-C １－４ Alkyl group, or C １－４ is an alkylamino group, n is 0, At least one of W', Y', Z', and V' is nitrogen (N); W' is nitrogen (N) or carbon (C); Y', Z', and V' are each independently nitrogen (N), sulfur (S), oxygen (O), or carbon (C); R' １ , R' ２ and R' ３ are each independently hydrogen, C １－４ Alkyl group, C １－４ Alkoxy group, -(CH2)-O-C １－４ Alkyl group, -(CH2CH2)-O-C １－４ Alkyl groups, 【Chemistry 3】 , or C １－４ is an alkylamino group, Said C １－４ Alkyl group, C １－４ Alkoxy group, -(CH2)-O-C １－４ Alkyl group, -(CH2CH2)-O-C １－４ Alkyl group, R ４ ', n', or C １－４ The alkylamino group is as defined in claim 1, M, a, X １ , b, c, X ２ and d is as defined in claim 1.

3. The compound of Chemical Formula 2 is a compound selected from the group consisting of compounds of Chemical Formulas 3 to 22 below. The transition metal complex or salt thereof according to claim 2: [Chemical formula 3] 【Chemistry 4】 [Chemical formula 4] 【Chemistry 5】 [Chemical formula 5] 【Chemistry 6】 [Chemical formula 6] 【Chemistry 7】 [Chemical formula 7] 【Chemistry 8】 [Chemical formula 8] 【Chemistry 9】 [Chemical formula 9] 【Chemistry 10】 [Chemical formula 10] 【Chemistry 11】 [Chemical formula 11] 【Chemistry 12】 [Chemical formula 12] 【Chemistry 13】 [Chemical formula 13] 【Chemistry 14】 [Chemical formula 14] 【Chemistry 15】 [Chemical formula 15] 【Chemistry 16】 [Chemical formula 16] 【Chemistry 17】 [Chemical formula 17] 【Chemistry 18】 [Chemical formula 18] 【Chemistry 19】 [Chemical formula 19] 【Chemistry 20】 [Chemical formula 20] 【Chemistry 21】 [Chemical formula 21] 【Chemistry 22】 , and [Chemical formula 22] 【Chemistry 23】

4. An oxidation-reduction polymer represented by a compound of the following chemical formula 26 or 27: [Chemical formula 26] 【Chemical 27】 , and [Chemical formula 27] 【Chemistry 28】 In the above formula, m and o each represent an integer selected from 10 to 600; M, L, a, X １ and X ２ are each as defined in claim 1.

5. The compound of formula 26 or formula 27 is a compound selected from the group consisting of compounds of formulas 28 to 43 below. [Chemical formula 28] 【Chemical 29】 [Chemical formula 29] 【Chemistry 30】 [Chemical formula 30] 【Chemistry 31】 [Chemical formula 31] 【Chemistry 32】 [Chemical formula 32] 【Chemical 33】 [Chemical formula 33] 【Chemical 34】 [Chemical formula 34] 【Chemistry 35】 [Chemical formula 35] 【Chemical 36】 [Chemical formula 36] 【Chemical 37】 [Chemical formula 37] 【Chemical 38】 [Chemical formula 38] 【Chemical Formula 39】 [Chemical formula 39] 【Chemistry 40】 [Chemical formula 40] 【Chemistry 41】 [Chemical formula 41] 【Chemistry 42】 [Chemical formula 42] 【Chemistry 43】 , and [Chemical formula 43] 【Chemistry 44】 In the above formula, m and o are each defined as in claim 4.

6. The oxidation-reduction polymer according to claim 4, further comprising a crosslinkable functional group and represented by the following formula 46 or 47: [Chemical formula 46] 【Chemistry 47】 , and [Chemical formula 47] 【Chemistry 48】 A Ｄ are primary and secondary amine groups, ammonium groups, halogen groups, epoxy groups, azide groups, acrylate groups, alkenyl groups, alkynyl groups, thiol groups, isocyanates, alcohol groups, silane groups, and 【Chemistry 49】 is one selected from the group consisting of The R ５ ' is hydrogen or a substituted or unsubstituted C １－４ is alkyl, n″ is an integer selected from 1 to 4, q is an integer selected from 1 to 10; m, o, and p each represent an integer selected from 10 to 600; M, L, a, X １ and X ２ are each as defined in claim 1.

7. The compound of the formula 46 or 47 is a compound selected from the group consisting of compounds of the following formulas 48 to 59. The oxidation-reduction polymer according to claim 6, [Chemical formula 48] 【Chemistry 50】 [Chemical formula 49] 【Chemistry 51】 [Chemical formula 50] 【Chemistry 52】 [Chemical formula 51] 【Chemistry 53】 [Chemical formula 52] 【Chemical 54】 [Chemical formula 53] 【Chemistry 55】 [Chemical formula 54] 【Chemistry 56】 [Chemical formula 55] 【Chemistry 57】 [Chemical formula 56] 【Chemistry 58】 [Chemical formula 57] 【Chemical Formula 59】 [Chemical formula 58] 【Chemistry 60】 , and [Chemical formula 59] 【Chemistry 61】 In the above formula, m, o and p are as defined in claim 6.

8. An electrochemical biosensor comprising the transition metal complex or a salt compound thereof according to any one of claims 1 to 3; or the oxidation-reduction polymer according to any one of claims 4 to 7.

9. The electrochemical biosensor of claim 8 , wherein the electrochemical biosensor is insertable into a body.

10. an enzyme capable of oxidizing-reducing a liquid biological sample, and 8. A sensing film for an electrochemical biosensor, comprising the transition metal complex or a salt compound thereof according to claim 1, or the oxidation-reduction polymer according to claim 4 as an electron transfer mediator.

11. The sensing membrane according to claim 10, wherein the enzyme is at least one selected from the group consisting of dehydrogenase, oxidase, and esterase.

12. 12. The sensing membrane according to claim 11, wherein the enzyme is 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.

13. 12. The sensing membrane according to claim 11, further comprising one or more cofactors selected from the group consisting of flavin adenine dinucleotide (FAD), nicotinamide adenine dinucleotide (NAD), and pyrroloquinoline quinone (PQQ).

14. The sensing membrane of claim 10 further comprising carbon nanotubes.

15. The sensing membrane of claim 10 , wherein the liquid biological sample is one or more selected from the group consisting of tissue fluid, blood, cells, plasma, serum, urine, cyst fluid, and saliva of a patient.

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