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US-12617793-B2 - Organic electron transfer mediator and device comprising same

US12617793B2US 12617793 B2US12617793 B2US 12617793B2US-12617793-B2

Abstract

The present invention relates to a novel organic electron-transfer mediator showing an excellent oxidation-reduction potential and a device such as an electrochemical biosensor having improved performance comprising the same.

Inventors

  • Hyunseo Shin
  • Young Jea KANG
  • Seok-Won Lee
  • Bongjin Moon
  • Myeonghwa JEONG
  • Sangeun YOON

Assignees

  • I-SENS, INC.
  • SOGANG UNIVERSITY RESEARCH & BUSINESS DEVELOPMENT FOUNDATION

Dates

Publication Date
20260505
Application Date
20201228
Priority Date
20191226

Claims (17)

  1. 1 . An organic electron-transfer mediator having the structure of Chemical formula 1 below: in the formula, R is —H, —F, —Cl, —Br, —I, —NO 2 , —CN, —CO 2 H, —SO 3 H, —NHNH 2 , —SH, —OH, —NR 1 R 2 , an unsubstituted or substituted alkyl group having 1 to 6 carbon atoms, an unsubstituted or substituted alkenyl group having 24 to 6 carbon atoms, or an unsubstituted or substituted aryl group having 6 to 10 carbon atoms, and the R 1 and R 2 may be each independently H, alkyl having 1 to 3 carbon atoms, or —COOR 3 , and the R 3 may be alkyl having 1 to 6 carbon atoms, L (linker) may be one or more selected from the group consisting of a bond, a substituted or unsubstituted alkyl group having 1 to 20 carbon atoms, a substituted or unsubstituted alkenyl group having 2 to 20 carbon atoms, a substituted or unsubstituted ethylene oxide group having 2 to 50 carbon atoms, a substituted or unsubstituted ethylene amine group having 2 to 50 carbon atoms, a substituted or unsubstituted alkoxy group having 1 to 30 carbon atoms, a substituted or unsubstituted aryl or aryloxy group having 6 to 30 carbon atoms, or a substituted or unsubstituted heteroaryl group or heteroaryloxy group having 5 to 30 carbon atoms.
  2. 2 . The organic electron-transfer mediator according to claim 1 , wherein the unsubstituted alkyl group having 1-20 carbon atoms in the L is one or more kinds selected from the group consisting of a methyl group, an ethyl group, a propyl group, an isopropyl group, an n-butyl group, a sec-butyl group, a t-butyl group, a pentyl group, a hexyl group, a cyclohexyl group, a heptyl group, an octyl group and a decane group; and the substituted or unsubstituted ethylene oxide group having 2 to 50 carbon atoms is one or more kinds selected from the group consisting of ethylene oxide groups in which the number of n in (—OCH 2 CH 2 —) n is 1-20; and the substituted or unsubstituted ethylene amine group having 2 to 50 carbo atoms is one or more kinds selected from the group consisting of ethylene amine groups in which the number of n in (—NHCH 2 CH 2 —) n is 1-20; and the substituted or unsubstituted alkoxy group having 1 to 30 carbon atoms may be one selected from the group consisting of methoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, pentanoxy, hexanoxy, heptanoxy, octanoxy, decanoxy, alkyl-decanoxy, dodecanoxy, alkyl-dodecanoxy, undecanoxy, alkyl-undecanoxy, allyloxy, cycloalkyloxy and cyclohexyloxy; and the substituted or unsubstituted aryl or aryloxy having 6 to 30 carbo atoms is one selected from the group consisting of a phenyl group, a tolyl group, a naphthalene group, a phenanthrene group, an alkyl phenyl group and a phenyloxy group, a benzyloxy group, a tolyloxy group, a naphthalene oxy group, a phenanthrene oxy group, and an alkoxyphenyl group; and the substituted or unsubstituted heteroaryl group or heteroaryloxy group having 5 to 20 carbon atoms is one selected from the group consisting of furyl, thiophenyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, thiadiazolyl, isothiazolyl, isooxazolyl, oxazolyl, oxadiazolyl, triazinyl, tetrazinyl, trazolyl, tetrazolyl, furazanyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, heteroaryl, benzofuranyl, benzothiophenyl, isobenzofuranyl, benzoimidazolyl, benzothiazolyl, benzoisothiazolyl, benzoixoazolyl, benzooxazolyl, isoindolyl, indolyl, indazolyl, benzothiadiazolyl, quinolyl, isoquinolyl, quinoxalinyl, carbazolyl, phenanthridinyl and benzodioxolyl.
  3. 3 . The organic electron-transfer mediator according to claim 1 , wherein the R is —H, —F, —Cl, —Br, —I, —NO 2 , —CN, —CO 2 H, —SO 3 H, —NHNH 2 , —SH, —OH, —NR 1 R 2 , an unsubstituted or substituted alkyl group having 1 to 3 carbon atoms, an unsubstituted or substituted alkenyl group having 2 to 3 carbon atoms, or a phenyl group, and the R 1 and R 2 is each independently H or Boc (t-butoxycarbonyl).
  4. 4 . The organic electron-transfer mediator according to claim 1 , wherein the L is one or more kinds selected from the group consisting of a bond, a substituted or unsubstituted alkylene having 1 to 8 carbon atoms, a substituted or unsubstituted alkenyl group having 2 to 6 carbon atoms, a substituted or unsubstituted ethylene oxide group having 2 to 6 carbon atoms, a substituted or unsubstituted ethylene amine group having 2 to 6 carbon atoms, a substituted or unsubstituted alkoxy group having 1 to 6 carbon atoms, a substituted or unsubstituted aryl or aryloxy group having 6 to 10 carbon atoms, or a substituted or unsubstituted heteroaryl group or heteroaryloxy group having 5 to 12 carbon atoms.
  5. 5 . The organic electron-transfer mediator according to claim 1 , wherein the -L-R is one selected from the following structures: H, —CH 2 CH 2 CH 2 SO 3 H, —CH 2 CH 2 CH 2 CH 2 CH 2 —NH(Boc), —CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 —NH(Boc), —CH 2 CH 2 —OH, —CH 2 CH 2 —Cl, —CH 2 CH 2 CH 2 CH 2 CH 2 —OH, —CH 2 CH 2 OCH 2 CH 2 —OH, and —CH 2 —CH═CH 2 .
  6. 6 . The organic electron-transfer mediator according to claim 1 , wherein the organic electron-transfer mediator having the structure of Chemical formula 1 is represented by any one structure of the following Chemical formulas 2 to 17:
  7. 7 . A method for preparation of an organic electron-transfer mediator of Chemical formula 2 below comprising i) reacting the compound of Chemical formula 18 below with 1,3-propanesultone to obtain the compound of Chemical formula 19 below; and ii) reacting the compound of Chemical formula 19 obtained in the i) with alloxan monohydrate and boric acid to obtain the compound of Chemical formula 2 below.
  8. 8 . The method for preparation according to claim 7 , wherein the amount of the 1,3-propanesultone used is 1.0 to 2.0 equivalents based on the compound of Chemical formula 18, and the amount of the alloxan monohydrate used is 1.0 to 1.5 equivalents based on the compound of Chemical formula 19.
  9. 9 . The method for preparation according to claim 7 , wherein the reaction temperature is 50 to 120° C. and the reaction time is 2 days to 4 days in the i), and the reaction temperature is 30 to 80° C. and the reaction time is 2 hours or more in the ii).
  10. 10 . A method for preparation of an organic electron-transfer mediator according to Chemical formula 1 comprising, i) reacting 1,2-dibromo-4,5-difluorobenzene of Chemical formula 20 below with polymethylhydrosiloxane, and then reacting with Zn(CN)2 under tris(dibenzylideneacetone)dipalladium(0) {Pd2(dba)3} and 1,1′-bis(diphenylphosphino)ferrocene (DPPF) to obtain the compound of Chemical formula 21 below; ii) reacting the compound of Chemical formula 21 obtained in the i) with ammonia water to obtain the compound of Chemical formula 22; iii) reacting the compound of Chemical formula 22 obtained in the ii) with the compound of Chemical formula 23 to obtain the compound of Chemical formula 24; and iv) reacting the compound of Chemical formula 24 obtained in the iii) with alloxan monohydrate and boric acid to obtain the compound of Chemical formula 1: in the formulas, L and R are same as defined in claim 1 .
  11. 11 . An oxidation-reduction polymer, comprising the organic electron-transfer mediator according to any one claim of claim 1 to claim 6 , and a polymer backbone selected from the group consisting of poly(vinylpyridine) (PVP), poly(vinylimidazole) (PVI) and poly allyl glycidyl ether (PAGE).
  12. 12 . The oxidation-reduction polymer according to claim 11 , b represented by Chemical formula 25: in the formula, X is 5 to 30.
  13. 13 . A device comprising the organic electron-transfer mediator according to claim 1 .
  14. 14 . The device according to claim 13 , wherein the device is an electrochemical biosensor.
  15. 15 . The device according to claim 13 , wherein the device is insertable.
  16. 16 . A sensing layer for an electrochemical biosensor comprising an enzyme capable of conducting oxidation reduction for a liquid biological sample; and the organic electron-transfer mediator according to claim 1 .
  17. 17 . The sensing layer according to claim 16 , wherein the enzyme comprises one or more kinds of oxidoreductases selected from the group consisting of dehydrogenase, oxidase and esterase; or one or more kinds of oxidoreductases selected from the group consisting of dehydrogenase, oxidase and esterase, and one or more kinds of cofactors selected from the group consisting of flavin adenine dinucleotide (FAD), nicotinamide adenine dinucleotide (NAD), and pyrroloquinoline quinone (PQQ).

Description

TECHNICAL FIELD The present invention relates to a novel organic electron-transfer mediator and an electrochemical biosensor comprising the same. BACKGROUND ART The operation principle of a glucose sensor is as FIG. 1. Specifically, glucose is oxidized by glucose oxidase to gluconic acid, and a reduced intermediate that receives electrons transfers electrons to an electrode. As a result, the flow of electrons generated by the reaction between blood glucose and enzyme is converted into an electrical signal, so that the blood glucose concentration can be known. A representative glucose dehydrogenase used in a blood glucose measurement strip is glucose dehydrogenase-flavin adenine dinucleotide (FAD-GDH). An electron-transfer mediator capable of helping movement of electrons between FAD-GDH and an electrode is required, and Fe(CN)6−3 is known as an appropriate compound because it is easily soluble in water, is inexpensive and has high sensitivity. However, due to low affinity of the active site, the rate constant (k2=kcat/kM) of FAD-GDH and Fe(CN)3 is as low as about 1×103 M−1s−1, so there is a limitation in that the reactivity between the two is not good. On the other hand, an osmium-based complex having a high rate constant is used as an electron-transfer mediator. When a ligand of the complex is modified, it can be adjusted to have an appropriate electrochemical potential, so it is very useful in terms of usability, but as the osmium metal is very expensive, it is not suitable for use in a disposable blood glucose measurement strip. Therefore, research on organic compounds as inexpensive and sustainable alternatives is in progress. Although the characteristics of various organic electron-transfer mediators including a naphthoquinone/phenanthrenequinone derivative, there is still a need for development of organic-based materials that can be used as such electron-transfer mediator. DISCLOSURE Technical Problem Under these circumstances, the present inventors have paid attention to oxidation reduction reaction of a phenothiazine organic electron transfer mediator. When the oxidation reduction reaction occurs in the corresponding phenothiazine organic electron transfer mediator as Reaction formula 1 below, the oxidation reduction potential is known as −0.1 V compared to the Ag/AgCl reference electrode when measured in an aqueous solution (See International Patent Publication No. WO 2008/036516, See U.S. Pat. No. 5,520,786). An important factor that the glucose sensor has poor accuracy in the hypoglycemic section (low response current section) is that there is a relatively high background current in this section. This background current occurs because there is a pathway for electrons from the surrounding environment in addition to the electrons from the enzyme. Therefore, in order to minimize this background current, the path through which the electron-transfer mediator receives electrons from the surrounding environment must be blocked, and for this purpose, it is optimal that the range of the standard reduction potential of the electron-transfer mediator has a value between about −0.2˜0.1 V compared to the Ag/AgCl reference electrode when measured in an aqueous solution. When measured in an organic solvent (CH3CN, DMSO, etc.), it may exhibit a value between −0.4˜0.1 V compared to the Ag/AgCl reference electrode. [Reaction Formula 1] Oxidation reduction reaction of phenothiazine organic electron transfer mediator On the other hand, the oxidation reduction potential of a flavin [or isoalloxazine]derivative such as Reaction formula 2 is known as −0.46 V compared to the Ag/AgCl reference electrode. This compound has a similar oxidation reduction potential to FAD, a derivative of FAD, well known as an oxidation reduction coenzyme, but it does not show an optimal oxidation reduction potential as an electron-transfer mediator for a glucose sensor. [Reaction Formula 2] Structure of riboflavin and flavin adenosine dinucleotide (FAD) and oxidation reduction reaction of derivatives thereof (isoalloxazine) Considering the oxidation reduction potential of the reported FAD-FDAH2 derivatives, it can be seen that the potential of the derivatives having an electron withdrawing substituent is shifted in a more positive direction (Reference J. Am. Chem. Soc. 1998, 120, 2251-2255) [Reaction formula 3]. According to this document, when electron withdrawing groups at positions 7 and 8 of the flavin framework, F, Cl, CN and the like are substituted, the oxidation reduction potential shifts in a more positive direction. However, when it is measured in an aqueous solution in the flavin derivatives described in this document, there is no compound having an optimal oxidation reduction potential (−0.2˜0.1V) of a glucose electron-transfer mediator. It can be expected to have a desired oxidation reduction potential value only when it has a CN group at least at the 7th and 8th positions or a substituent with a stronger electron withdrawing g