JP7421268B2 - new mediator - Google Patents

Description

The present invention relates to the use of a phenylenediamine compound as a mediator, an electrode modifier containing the compound, an electrode containing the compound, an enzyme sensor containing the electrode, and a battery. The present invention also relates to an electrochemical measurement using a phenylenediamine compound and an oxidoreductase, a composition containing a phenylenediamine compound and an oxidoreductase, and an electrode containing a phenylenediamine compound.

In an enzyme electrode, a mediator is used as a mediating substance for transferring electrons generated by an enzyme-mediated redox reaction to an electrode. For efficient electron transfer from the enzyme to the electrode, it is desirable that the mediator be localized near the electrode. Therefore, various methods are used to immobilize the mediator on the electrode, such as a method of chemically bonding the electrode and the mediator, and a method of polymerizing the mediator itself. However, the chemical bonding method has limitations on the side chain functional groups of the mediator, and the method of polymerizing the mediator itself may change the redox potential of the mediator, so conventional methods lacked versatility. . It has also been reported that treating a glassy carbon electrode with a strong acid activates the functional groups on the surface and adsorbs and fixes mediators such as quinones, but this has hardly been put to practical use in industry.

Furthermore, both methods require complicated steps to immobilize the mediator on the electrode, resulting in high costs. Therefore, there has been a desire for an easily usable electronic mediator that does not require complicated steps for immobilization on electrodes.

Patent Document 1 (JP-A-7-234201) describes a p-phenylenediamine compound as an electron mediator used in an electrochemical measurement method. The disclosed p-phenylenediamine compound is a p-phenylenediamine derivative having one or more groups selected from the group consisting of a hydroxyl group, a mercapto group, a carboxyl group, a phosphonooxy group, and a sulfo group.

Patent Document 2 (International Publication No. 2004/011929 pamphlet) discloses that carbon particles are treated with an acid to activate their surfaces, and then N,N'-diphenyl-p-phenylenediamine (DPPD) is added to the acid-treated carbon powder. ), and the immobilization of this carbon powder on a carbon electrode, and the detection of hydrogen sulfide and thiol in a solution using this electrode.

Patent Document 3 (Japanese Patent Publication No. 2007-526474) describes an electrode containing carbon derivatized with N,N'-diphenyl-p-phenylenediamine, and a pH sensor using the electrode.

Patent Document 4 (JP 2008-185534) discloses 2,3,5,6-tetramethyl-1,4-phenylenediamine or N,N-dimethyl-p-phenylenediamine, which is a phenylenediamine compound, as a mediator, and The ethanol measurement using this is described.

Patent Document 5 (JP 2016-042032) describes N,N,N',N'-tetramethyl-1,4-phenylenediamine as a mediator and glucose measurement using the same.
N-isopropyl-N'-phenyl-p-phenylenediamine (IPPD), N,N'-diphenyl-p-phenylenediamine (DPPD), N-(1,3-dimethylbutyl)-N'-phenyl-p- Phenyl diamine (6PPD) is known as an anti-aging agent for rubber (Non-Patent Document 2, Journal of the Japan Rubber Association, Vol. 82, No. 2, 2009, p. 45-49).

Non-Patent Document 1 (Analyst, 2003, 128, 473-479) discloses that after acid-treating carbon powder with 0.1 M hydrochloric acid to activate the carbon surface, DPPD is added to the acid-treated carbon powder, and this carbon We report the immobilization of powder onto a carbon electrode (basal plane pyrolytic graphite electrode, BPPG electrode) and the detection of sulfides using this electrode. US Pat. No. 6,001,300 describes a flow battery.

JP 7-234201 International Publication No. 2004/011929 pamphlet Special table 2007-526474 (International Publication No. 2005/085825 pamphlet) JP2008-185534 JP2016-042032 JP2019-003928

Analyst, 2003, 128, 473-479 Japan Rubber Association Journal, Volume 82, No. 2, 2009, p. 45-49

The present invention aims to provide a new mediator that can at least partially solve the above problems.

As a result of our earnest efforts to solve the above problems, the present inventors have surprisingly found that phenylenediamine compounds can be adsorbed onto the electrode surface without the need for special treatment such as acid treatment. They also discovered that this compound can function as a mediator, and completed the present invention. As far as the present inventors know, it has not been reported that IPPD, DPPD, and 6PPD are mediators that adsorb to electrodes, and this is a surprising finding. Additionally, the present inventors surprisingly discovered a further phenylenediamine compound that can be adsorbed onto the electrode surface without requiring special treatment such as acid treatment.

［式中、
R¹は、-NR⁷R⁸、-N=N-R⁹、又は-N⁺≡Nであり、
R²は、-NR¹⁰R¹¹又は、-N=N-R¹²であり、
R⁷及びR⁸は、それぞれ独立に、水素、場合により1以上のX又はVにより置換されてもよい、直鎖又は分枝鎖のC_1-7アルキル、C_1-7アルケニル、C_1-7アルキニル、C_3-9シクロアルキル、フェニル、1-ナフチル、2-ナフチル、アントラセニル、フェナントレニル、アセチル、カルボキシ、フラニルホルミル、ピラゾリルホルミル、1-メチル-1H-ピラゾール-5-イルホルミル、9,9-ジメチルフルオレン-2-イル、-N⁺≡N、-N=N-フェニル、ベンジル、

であり、
R¹⁰は、水素、場合により1以上のX又はVにより置換されてもよい、直鎖又は分枝鎖のC_1-7アルキル、C_1-7アルケニル、C_1-7アルキニル、C_3-9シクロアルキル、フェニル、1-ナフチル、2-ナフチル、アントラセニル又はフェナントレニル、であり、
R³、R⁴、R⁵及びR⁶は、それぞれ独立に、水素、場合により1以上のYにより置換されてもよい、直鎖又は分枝鎖のC_1-7アルキル、C_1-7アルケニル、C_1-7アルキニル、C_1-7アルコキシ、ハロ、ニトロ、シアノ、カルボキシ、スルホ、ヒドロキシ又はアミノであり、
或いは、R³及びR⁴、又は、R⁵及びR⁶は、それらを含有するベンゼン環と一緒になって、場合により1以上のオキソ、X又はWにより置換されてもよい、ベンゼン環、又は

を形成し、但し＊はR³が結合する炭素原子に結合し、＊＊はR⁴が結合する炭素原子に結合するか、又は、＊はR⁵が結合する炭素原子に結合し、＊＊はR⁶が結合する炭素原子に結合し、
R¹¹は、場合により１以上のX又はZにより置換されてもよい、フェニル、1-ナフチル、2-ナフチル、アントラセニル及びフェナントレニルからなる群より選択され、
R⁹は、水素、場合により１以上のXにより置換されてもよい、フェニル、1-ナフチル、2-ナフチル、アントラセニル及びフェナントレニルからなる群より選択され、
R¹²は、場合により１以上のX、W、イソチオシアネート、又はハロスルホニルにより置換されてもよい、フェニル、1-ナフチル、2-ナフチル、アントラセニル、フェナントレニル、及び

からなる群より選択され、
ここでVは、場合によりC1-7アルキルに置換されてもよい、-O-アクリロイル、アセチルアミノ、又はフェニルであり、
Wは、場合により1以上のアミノ、C1-7アルキル、アミノアルキルにより置換されてもよい、D若しくはL-アラニルスルホニル、D若しくはL-バリルスルホニル、D若しくはL-ロイシルスルホニル、D若しくはL-メチオニルスルホニル、D若しくはL-プロリルスルホニル、D若しくはL-トリプトフィルスルホニル、D若しくはL-グリシルスルホニル、D若しくはL-システイニルスルホニル、D若しくはL-イソロイシルスルホニル、D若しくはL-フェニルアラニルスルホニル、D若しくはL-チロシルスルホニル、D若しくはL-セリルスルホニル、D若しくはL-トレオニルスルホニル、D若しくはL-アスパラギニルスルホニル、D若しくはL-グルタミルスルホニル、D若しくはL-アルギニルスルホニル、D若しくはL-ヒスチジルスルホニル、D若しくはL-リジルスルホニル、D若しくはL-アスパラギルスルホニル、D若しくはL-グルタミニルスルホニル、-C(=O)-O-スクシンイミジル、アセチル、トリフルオロアセチル、ベンゾイルアミノ、-N=N-フェニル、フェニルアミノ、ジアミノフェニルアゾフェニル、又は、場合により１以上のXにより置換されてよいフェニルアゾ、場合により１以上のYにより置換されてもよいナフチルアゾ、アセチルアミノ、

であり、
Xは、場合により1以上のハロ、アミノ、シアノ、カルボキシ、カルボニル、アルコキシ、アルキルアミノ、ニトロソ、ニトロ及びスルホからなる群より選択される置換基により置換されてもよい、直鎖又は分枝鎖の、C_1-7アルキル、C_1-7アルケニル、C_1-7アルキニル、C_1-7アルコキシ、ハロ、ヒドロキシ、ニトロ、カルボキシ、シアノ、スルホ、アミノ又はアルキルアミノであり、
Yはハロ、アミノ、シアノ、カルボキシ、カルボニル、ヒドロキシ、アルコキシ及びスルホからなる群より選択され、
Zは-SO₂-CH=CH₂、又は-SO₂-C₂H₄-O-SO₃H、4,6-ジクロロトリアジン-2-イルアミノである］
を有する化合物、またはその塩、無水物若しくは溶媒和物である、
実施形態１に記載の電極改変剤、
実施形態２に記載の電子伝達促進剤、
実施形態３～６のいずれかに記載の電池、
実施形態７、９又は１０に記載の組成物、
実施形態８～１１のいずれかに記載の電極、又は
実施形態１２に記載の酵素センサー。
[１４] 化合物が、式Ia若しくはIIaの構造

［式中、
R^1aは、-NR^7aR^8a、-N=N-R^9a、又は-N⁺≡Nであり、
R^2aは、-NR^10aR^11a、又は-N=N-R^12aであり、
R^7a及びR^8aは、それぞれ独立に、水素、場合により1以上のXaにより置換されてもよい、直鎖又は分枝鎖のC_1-6アルキル、C_1-6アルケニル、C_1-6アルキニル、C_3-9シクロアルキル、フェニル、1-ナフチル、2-ナフチル、アントラセニル、又はフェナントレニルであり、
R¹⁰は、水素、場合により1以上のXaにより置換されてもよい、直鎖又は分枝鎖のC_1-6アルキル、C_1-6アルケニル、C_1-6アルキニル、C_3-9シクロアルキル、フェニル、1-ナフチル、2-ナフチル、アントラセニル又はフェナントレニルであり、
R^3a、R^4a、R^5a及びR^6aは、それぞれ独立に、水素、場合により1以上のYにより置換されてもよい、直鎖又は分枝鎖のC_1-6アルキル、C_1-6アルケニル、C_1-6アルキニル、C_1-6アルコキシ、ハロ、ニトロ、シアノ、カルボキシ、スルホ、ヒドロキシ又はアミノであり、
或いは、R^3a及びR^4a、又は、R^5a及びR^6aは、ベンゼン環、又は

を形成し、但し＊はR^3aが結合する炭素原子に結合し、＊＊はR^4aが結合する炭素原子に結合するか、又は、＊はR^5aが結合する炭素原子に結合し、＊＊はR^6aが結合する炭素原子に結合し、
R^11aは、場合により１以上のXaにより置換されてもよい、フェニル、1-ナフチル、2-ナフチル、アントラセニル及びフェナントレニル、からなる群より選択され、
R^9aおよびR^12aは、それぞれ独立に、水素、場合により１以上のXaにより置換されてもよい、フェニル、1-ナフチル、2-ナフチル、アントラセニル及びフェナントレニル、からなる群より選択され、
Xaは、場合により1以上のハロ、アミノ、シアノ、カルボキシ、カルボニル、ヒドロキシ、アルコキシ、アルキルアミノ、ニトロソ、ニトロ及びスルホからなる群より選択される置換基により置換されてもよい、直鎖又は分枝鎖の、C_1-6アルキル、C_1-6アルケニル、C_1-6アルキニル、C_1-6アルコキシ、ハロ、ヒドロキシ、ニトロ、カルボキシ、シアノ、スルホ、アミノ又はアルキルアミノであり、
Yはハロ、アミノ、シアノ、カルボキシ、カルボニル、アルコキシ及びスルホからなる群より選択される］
を有する、又は、式Ib若しくはIIｂの構造

［式中、
R^7b、R^8b及びR^10bは、それぞれ独立に、水素、場合により1以上のXbにより置換されてもよい、直鎖又は分枝鎖のC_1-6アルキル、C_1-6アルケニル、C_1-6アルキニル、C_3-9シクロアルキル、フェニル、1-ナフチル、2-ナフチル、アントラセニル又はフェナントレニル、であり、R^3b、R^4b、R^5b及びR^6bは、それぞれ独立に、水素、場合により1以上のYにより置換されてもよい、直鎖又は分枝鎖のC_1-6アルキル、C_1-6アルケニル、C_1-6アルキニル、C_1-6アルコキシ、ハロ、ニトロ、シアノ、カルボキシ、スルホ又はアミノであり、
R^11bは、場合により１以上のXbにより置換されてもよい、フェニル、1-ナフチル、2-ナフチル、アントラセニル及びフェナントレニル、からなる群より選択され、
ここでXbは、場合により1以上のハロ、アミノ、シアノ、カルボキシ、カルボニル、アルコキシ及びスルホからなる群より選択される置換基により置換されてもよい、直鎖又は分枝鎖の、C_1-6アルキル、C_1-6アルケニル、C_1-6アルキニル、C_1-6アルコキシ、ハロ、ヒドロキシ、ニトロ、カルボキシ、シアノ、スルホ又はアミノであり、
Yはハロ、アミノ、シアノ、カルボキシ、カルボニル、アルコキシ及びスルホからなる群より選択される］
を有する化合物、またはその塩、無水物若しくは溶媒和物である、実施形態１３に記載の電極改変剤、電子伝達促進剤、電池、組成物、電極、又はセンサー。
[１５] 化合物が、

からなる群より選択される、実施形態１３又は１４に記載の電極改変剤、電子伝達促進剤、電池、組成物、電極又はセンサー。
[１６] 実施形態１に記載の電極改変剤又は実施形態２に記載の電子伝達促進剤を用いる工程を含む、電池の製造方法。
[１７] 前記電極改変剤、又は電子伝達促進剤を、電池の電極と接触させる工程を含む、実施形態１６に記載の方法。
[１８] 実施形態３～６及び１３～１５のいずれかに記載の電池を用いる、発電方法。
[１９] 実施形態７、９、１０及び１３～１５のいずれかに記載の組成物、
実施形態８～１１、及び１３～１５のいずれかに記載の電極、又は
実施形態１２～１５のいずれかに記載のセンサー
を用いる、電気化学的測定方法。
[２０] 実施形態１及び１３～１５のいずれかに記載の電極改変剤、
実施形態２及び１３～１５のいずれかに記載の電子伝達促進剤、又は
実施形態７、９、１０及び１３～１５のいずれかに記載の組成物、
を電極に接触させる工程を含む、電極を改変又は修飾する方法。
本明細書は本願の優先権の基礎となる日本国特許出願番号２０１８－０７８７０１号の開示内容を包含する。 The invention includes the following embodiments:
[1] An electrode modifying agent containing a compound that has the property of adsorbing to a non-acid-treated electrode without being bonded to a polymer or being polymerized.
[2] An electron transfer promoter containing a compound that has the property of being adsorbed to an unacid-treated electrode without being bonded to a polymer or polymerized.
[3] A battery comprising the electrode modifier according to Embodiment 1 or the electron transfer enhancer according to Embodiment 2.
[4] The battery according to Embodiment 3, wherein the compound is immobilized on an electrode of the battery.
[5] The battery according to Embodiment 3 or 4, which contains an oxidoreductase.
[6] The battery according to Embodiment 5, wherein the oxidoreductase is immobilized on the electrode.
[7] A composition comprising the electrode modifier according to Embodiment 1 or the electron transfer enhancer according to Embodiment 2.
[8] An electrode comprising the electrode modifier according to Embodiment 1 or the electron transfer enhancer according to Embodiment 2.
[9] The electrode according to Embodiment 8, which has an enzyme, or the composition according to Embodiment 7, which includes an enzyme.
[10] The electrode or composition according to Embodiment 9, wherein the enzyme is an oxidoreductase.
[11] The electrode according to Embodiment 10, in which an oxidoreductase is immobilized.
[12] A sensor comprising the electrode modifier according to Embodiment 1 or the electron transfer enhancer according to Embodiment 2, or an enzyme sensor having the electrode according to Embodiment 10 or 11.
[13] The compound has the structure of formula I or formula II,

[In the formula,
R ¹ is -NR ⁷ R ⁸ , -N=NR ⁹ , or -N ⁺ ≡N,
R ² is -NR ¹⁰ R ¹¹ or -N=NR ¹² ,
R ⁷ and R ⁸ are each independently hydrogen, a straight chain or branched C _1-7 alkyl, C _1-7 alkenyl, C _1- which may optionally be substituted with one or more X or V; ₇ alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, acetyl, carboxy, furanylformyl, pyrazolylformyl, 1-methyl-1H-pyrazol-5-ylformyl, 9,9 -dimethylfluoren-2-yl, -N ⁺ ≡N, -N=N-phenyl, benzyl,

and
R ¹⁰ is hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _3-9 optionally substituted by one or more X or V; cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl,
R ³ , R ⁴ , R ⁵ and R ⁶ are each independently hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl, optionally substituted with one or more Y; , C _1-7 alkynyl, C _1-7 alkoxy, halo, nitro, cyano, carboxy, sulfo, hydroxy or amino,
Alternatively, R ³ and R ⁴ or R ⁵ and R ⁶ together with the benzene ring containing them are optionally substituted with one or more oxo, X or W, or

, where * is bonded to the carbon atom to which R ³ is bonded, ** is bonded to the carbon atom to which R ⁴ is bonded, or * is bonded to the carbon atom to which R 5 is bonded, and ** is bonded to the carbon atom to which R ⁵ is bonded. is bonded to the carbon atom to which R ⁶ is bonded,
R ¹¹ is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X or Z;
R ⁹ is selected from the group consisting of hydrogen, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X;
R ¹² is phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, optionally substituted with one or more of X, W, isothiocyanate, or halosulfonyl;

selected from the group consisting of
Here, V is -O-acryloyl, acetylamino, or phenyl, which may be optionally substituted with C1-7 alkyl,
W is D or L-alanylsulfonyl, D or L-valylsulfonyl, D or L-leucylsulfonyl, D or L, optionally substituted with one or more amino, C1-7 alkyl, aminoalkyl -methionylsulfonyl, D or L-prolylsulfonyl, D or L-tryptophylsulfonyl, D or L-glycylsulfonyl, D or L-cysteinylsulfonyl, D or L-isoleuylsulfonyl, D or L-phenylalanylsulfonyl, D or L-tyrosylsulfonyl, D or L-serylsulfonyl, D or L-threonylsulfonyl, D or L-asparaginylsulfonyl, D or L-glutamylsulfonyl, D or L- Arginylsulfonyl, D or L-histidylsulfonyl, D or L-lysylsulfonyl, D or L-asparagylsulfonyl, D or L-glutaminylsulfonyl, -C(=O)-O-succinimidyl, acetyl, tri- fluoroacetyl, benzoylamino, -N=N-phenyl, phenylamino, diaminophenylazophenyl, or phenylazo optionally substituted by one or more X, naphthylazo optionally substituted by one or more Y, acetylamino,

and
X is a straight or branched chain, optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy, alkylamino, nitroso, nitro, and sulfo. is C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _1-7 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo, amino or alkylamino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, hydroxy, alkoxy and sulfo;
Z is -SO ₂ -CH=CH ₂ or -SO ₂ -C ₂ H ₄ -O-SO ₃ H, 4,6-dichlorotriazin-2-ylamino]
or a salt, anhydride or solvate thereof,
the electrode modifier according to Embodiment 1;
The electron transfer enhancer according to Embodiment 2,
The battery according to any one of embodiments 3 to 6,
The composition according to embodiment 7, 9 or 10,
The electrode according to any of embodiments 8 to 11 or the enzyme sensor according to embodiment 12.
[14] The compound has a structure of formula Ia or IIa

[In the formula,
R ^1a is -NR ^7a R ^8a , -N=NR ^9a , or -N ⁺ ≡N,
R ^2a is -NR ^10a R ^11a or -N=NR ^12a ,
R ^7a and R ^8a are each independently hydrogen, linear or branched C _1-6 alkyl, C _1-6 alkenyl, C _1-6 alkynyl, optionally substituted with one or more Xa; , C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, or phenanthrenyl,
R ¹⁰ is hydrogen, linear or branched C _1-6 alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _3-9 cycloalkyl, optionally substituted with one or more Xa , phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl,
R ^3a , R ^4a , R ^5a and R ^6a are each independently hydrogen, linear or branched C _1-6 alkyl, C _1-6 alkenyl, optionally substituted with one or more Y; , C _1-6 alkynyl, C _1-6 alkoxy, halo, nitro, cyano, carboxy, sulfo, hydroxy or amino,
Alternatively, R ^3a and R ^4a or R ^5a and R ^6a are benzene rings, or

, where * is bonded to the carbon atom to which R ^3a is bonded, ** is bonded to the carbon atom to which R ^4a is bonded, or * is bonded to the carbon atom to which R ^{5a is bonded, and ** is bonded to the carbon atom to which R 5a} is bonded. is bonded to the carbon atom to which R ^6a is bonded,
R ^11a is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more Xa;
R ^9a and R ^12a are each independently selected from the group consisting of hydrogen, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted with one or more Xa;
Xa is a linear or branched chain, optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, hydroxy, alkoxy, alkylamino, nitroso, nitro and sulfo. branched C _1-6 alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _1-6 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo, amino or alkylamino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo]
or has a structure of formula Ib or IIb

[In the formula,
R ^7b , R ^8b and R ^10b are each independently hydrogen, a linear or branched C _1-6 alkyl, C _1-6 alkenyl, C ₁ which may optionally be substituted with one or more Xb _-6 alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl, and R ^3b , R ^4b , R ^5b and R ^6b are each independently hydrogen, optionally 1 Straight chain or branched C _1-6 alkyl, C _1-6 alkenyl, C _{1-6 alkynyl, C 1-6 alkoxy} , halo, nitro, cyano, carboxy, sulfo, which may be substituted by the above Y or amino;
R ^11b is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more Xb;
Here, Xb is a linear or branched C 1- which may be optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo _{. 6} alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _1-6 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo or amino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo]
or a salt, anhydride, or solvate thereof, the electrode modifier, electron transfer promoter, battery, composition, electrode, or sensor according to Embodiment 13.
[15] The compound is

The electrode modifier, electron transfer enhancer, battery, composition, electrode or sensor according to Embodiment 13 or 14, selected from the group consisting of:
[16] A method for manufacturing a battery, including a step of using the electrode modifier according to Embodiment 1 or the electron transfer enhancer according to Embodiment 2.
[17] The method according to Embodiment 16, comprising the step of bringing the electrode modifier or electron transfer promoter into contact with an electrode of a battery.
[18] A power generation method using the battery according to any one of Embodiments 3 to 6 and 13 to 15.
[19] The composition according to any one of Embodiments 7, 9, 10 and 13 to 15,
An electrochemical measurement method using the electrode according to any one of Embodiments 8 to 11 and 13 to 15, or the sensor according to any one of Embodiments 12 to 15.
[20] The electrode modifier according to any one of Embodiments 1 and 13 to 15,
The electron transfer enhancer according to any one of Embodiments 2 and 13 to 15, or the composition according to any one of Embodiments 7, 9, 10, and 13 to 15,
A method of altering or modifying an electrode, the method comprising the step of contacting an electrode with an electrode.
This specification includes the disclosure content of Japanese Patent Application No. 2018-078701, which is the basis of the priority of this application.

Unlike conventional mediators such as p-phenylenediamine, the phenylenediamine compound of the present invention is adsorbed directly onto the electrode surface without requiring any special treatment such as acid treatment of the electrode or polymerization of the mediator itself. Therefore, it can be easily immobilized on an electrode. Such electrodes can also be used for electrochemical measurements. Further, such electrodes can be applied to batteries.

Cyclic voltammetry was performed using IPPD and GDH, and the results of plotting the sweep speed, I _Omax , and I _Rmax are shown. The results are shown using DPPD instead of IPPD. The results using 6PPD instead of IPPD are shown. The results are shown using p-phenylenediamine instead of IPPD. The results are shown using p-phenylenediamine instead of IPPD. The horizontal axis is the square root of the sweep speed. The results using BGLB instead of IPPD are shown. The results using TDPA instead of BGLB are shown. A cyclic voltammogram using IPPD is shown. The relationship between the final glucose concentration and the oxidation current value at 300 mV is shown when IPPD is used. The relationship between the final glucose concentration and the oxidation current value at 300 mV is shown when using DPPD. The relationship between the final glucose concentration and the oxidation current value at 300 mV when using 6PPD is shown. The relationship between the final glucose concentration and the oxidation current value at 300 mV when using FADGDH-AA and IPPD is shown. The relationship between the final glucose concentration and the oxidation current value at 300 mV is shown when GLD1 and IPPD are used. The relationship between the final glucose concentration and the oxidation current value at 300 mV when using GOD and IPPD is shown. The relationship between the final glucose concentration and the oxidation current value at 300 mV when using FPOX-CE IPPD is shown. A comparison of oxidation current values when +300 mV is applied using BGLB with FADGDH-AA is shown. A comparison of oxidation current values when +300 mV is applied using TDPA with FADGDH-AA is shown. The results of plotting the relationship between the final glucose concentration and the oxidation current value at +300 mV using IPPD with PQQ-GDH are shown. The results of plotting the relationship between the final glucose concentration and the oxidation current value at +300 mV using BGLB together with PQQ-GDH are shown. The response current before and after glucose addition is shown when NAD-GDH is used in the presence or absence of DPPD. The results of plotting the relationship between the final lactic acid concentration and the oxidation current value at +150 mV using IPPD with LOD are shown. The results of plotting the relationship between the final fructose concentration and the oxidation current value at +100 mV when FDH is used in the presence or absence of IPPD are shown. The current when no fructose was added was set to 0. The relationship between the final glucose concentration and the oxidation current value at 200 mV is shown when using an electrode with IPPD adsorbed and GDH immobilized. The oxidation current value at +100 mV is shown when an electrode modified using DPPD at a final concentration of 10 pM is placed in a solution containing FADGDH-AA and glucose. The current when no glucose was added was defined as 0.

(Phenylene diamine compound)
(Electrode with phenylenediamine compound adsorbed)
In certain embodiments, the invention provides phenylenediamine-based compounds. The compounds of the present invention also include azo compounds and diazonio compounds, but in this specification, when referring to a phenylenediamine compound or the phenylenediamine compound of the present invention, for convenience, it includes not only compounds having a phenylenediamine skeleton, but also compounds having a phenylenediamine skeleton. Azo compounds and diazonio compounds are also included. In another embodiment, the invention provides an electrode modifier comprising a phenylenediamine-based compound. This electrode modifying agent can be adsorbed onto the electrode surface and can modify the electron accepting properties of the electrode. In another embodiment, the present invention provides an electrode on which a phenylenediamine-based compound or an electrode modifier of the present invention is adsorbed. In another embodiment, the present invention provides a composition for electrochemical measurement comprising a phenylenediamine-based compound or an electrode modifier of the present invention. The composition may further include an enzyme. The enzyme can be an oxidoreductase. In another embodiment, the present invention provides an electrochemical measurement kit comprising a phenylenediamine compound or an electrode modifier of the present invention. In this specification, the electrode modifying agent is sometimes referred to as an electrode adsorbent.

The phenylenediamine compound of the present invention can be adsorbed onto the electrode surface by bringing it into contact with the electrode. At this time, special operations such as activating the electrode surface by acid treatment are not required in order to cause it to be adsorbed onto the electrode. In one embodiment, the property of the compound of the present invention to be adsorbed to an electrode refers to the property of the compound to be physically adsorbed to an electrode, and the electrode may be made of a material such as carbon, gold, or platinum. Furthermore, the present invention includes an embodiment in which the compound is adsorbed onto a primary material such as carbon powder or a carbon material, and then the primary material such as the carbon powder or carbon material is immobilized on an electrode. However, this is an explanation of the properties that the compound of the present invention has, and does not limit the method of using the compound. That is, in some embodiments, the present invention provides a method of adsorbing a compound of the present invention onto a primary material such as a carbon powder or carbon material, and then immobilizing the primary material such as the carbon powder or carbon material on an electrode.

In certain embodiments, the present invention provides a primary material, such as a carbon powder or carbon material, on which a compound of the present invention is adsorbed. This primary material, such as carbon powder or carbon material, can be further coated or applied to the electrode. Primary materials include, but are not limited to, carbon, platinum, gold, and the like. Examples of the carbon material include carbon black, carbon fiber, single-walled or multi-walled carbon nanotubes, graphene, Ketjenblack, and the like.

In one embodiment, the property of the compound of the present invention to adsorb to an electrode, carbon powder, or carbon material refers to the property of the compound to physically adsorb directly to the electrode, carbon powder, or carbon material; This does not refer to the property of bonding to an electrode, carbon powder, or carbon material by covalently bonding to a polymer or linker. In this specification, such a property is defined as a property of being adsorbed to an electrode without being bound to a polymer, or a property of being adsorbed to a carbon powder or carbon material without being bound to a polymer or without the compound being polymerized. There is a thing called nature. However, this is an explanation of the properties that the compound of the present invention has, and does not limit the method of using the compound. Thus, in certain embodiments, the present invention provides a method of bonding a compound of the present invention to an electrode, carbon powder, or carbon material by covalently bonding it to a polymer or linker. Furthermore, the fact that the compound is physically adsorbed directly to the electrode, carbon powder, or carbon material does not mean that the compound of the present invention itself (as a monomer) can be covalently bonded to the carbon powder or carbon material. In this specification, the term "polymer" refers to a polymer composed of, for example, 10 or more identical units.

In certain embodiments, the carbon powder or carbon material, carbon electrode, gold electrode, platinum electrode for adsorbing the compounds of the invention is not acid-treated. That is, in one embodiment, the acid-treated carbon powder or carbon material and carbon electrode are excluded from the carbon powder or carbon material on which the compound of the present invention is adsorbed.

In certain embodiments, compounds of the invention can be used in combination with enzymes. In certain embodiments, compounds of the invention can be used in combination with oxidoreductases. In certain embodiments, compounds of the invention can be used as electron transfer enhancers. In one embodiment, the phenylenediamine compound of the present invention functions as a mediator in a redox reaction catalyzed by an oxidoreductase. Examples of oxidoreductases include various oxidoreductases classified into EC Group 1, such as glucose oxidase, glucose dehydrogenase, amadoriase (also referred to as fructosyl peptide oxidase or fructosyl amino acid oxidase), peroxidase, galactose oxidase, bilirubin oxidase, Pyruvate oxidase, D- or L-amino acid oxidase, amine oxidase, cholesterol oxidase, choline oxidase, xanthine oxidase, sarcosine oxidase, D- or L-lactate oxidase (LOD), ascorbate oxidase, cytochrome oxidase, alcohol dehydrogenase, cholesterol dehydrogenase , aldehyde dehydrogenase, aldehyde oxidase, fructose dehydrogenase (FDH), sorbitol dehydrogenase, D- or L-lactate dehydrogenase, malate dehydrogenase, glycerol dehydrogenase, 17B hydroxysteroid dehydrogenase, estradiol 17B dehydrogenase, D- or L-amino acid dehydrogenase, glycerol Aldehyde 3-phosphate dehydrogenase, 3-hydroxysteroid dehydrogenase, diaphorase, catalase, glutathione reductase, cytochrome B5 reductase, adrenoxine reductase, cytochrome B5 reductase, adrenodoxin reductase, nitrate reductase, phosphate dehydrogenase, bilirubin oxidase, laccase, Examples include, but are not limited to, polyamine oxidase, formate dehydrogenase, pyranose oxidase, pyranose dehydrogenase, tauropine dehydrogenase, and the like. Alternatively, it may be a combination of multiple enzymes. Coenzymes for the above enzymes include nicotinamide adenine dinucleotide (NAD), nicotinamide adenine dinucleotide phosphate, flavin adenine dinucleotide (FAD), pyrroloquinoline quinone, and the like. That is, examples of glucose dehydrogenase (GDH) include FAD-dependent GDH, NAD-dependent GDH, and PQQ-dependent GDH. The activity of the above-mentioned oxidoreductases can be measured using various substrates, for example, by the method described in Methods in Enzymology (Volumes 1 to 602).

In this specification, functioning as a mediator means that the phenylenediamine compound participates in electron transfer. For example, in a system using an electrode, the phenylenediamine compound of the present invention receives electrons from an oxidoreductase and reduces the form, transfers electrons to the electrode, and returns to the oxidized form. From this point of view, the phenylenediamine compound of the present invention that functions as a mediator can also be referred to as an electron transfer mediator, an electron transfer promoter, or an electron mediator (also simply referred to as a mediator). In this specification, these terms are used synonymously.

In certain embodiments, the electrodes of the present invention exclude electrodes having acid-treated carbon powder or particles.

In certain embodiments, the oxidoreductase used with the electrode of the present invention may be immobilized on the electrode. That is, in this embodiment, the present invention provides an electrode on which an oxidoreductase is immobilized and an electrode modifying agent of the present invention is adsorbed. In one embodiment, the present invention provides an enzyme sensor comprising an electrode on which an oxidoreductase is immobilized and an electrode modifier of the present invention is adsorbed. In some embodiments, sensors are provided that include electrode modifiers or electron transfer enhancers of the invention.

The phenylenediamine-based compound of the present invention or the electrode modifier, electrode adsorbent, or electron transfer promoter containing the phenylenediamine-based compound can be adsorbed onto the electrode surface without requiring any special treatment. Therefore, in some embodiments, in order to produce a modified electrode, the phenylenediamine compound, electrode modifier, electrode adsorbent, or electron transfer promoter of the present invention may be adsorbed onto the electrode in advance. In another embodiment, when performing an electrochemical measurement, a measurement solution (composition) containing the phenylenediamine compound of the present invention, an electrode modifier, an electrode adsorbent or an electron transfer promoter, and an oxidoreductase is used. can be used. In this embodiment, when a composition comprising a phenylenediamine compound of the present invention, an electrode modifier, an electrode adsorbent or an electron transfer promoter, and an oxidoreductase is brought into physical contact with an electrode, the phenylenediamine of the present invention A system compound, an electrode modifier, an electrode adsorbent, or an electron transfer promoter is adsorbed to the electrode, and an electrode with modified electrochemical properties is produced in situ at the time of measurement.

In one embodiment, the electrode is not pretreated with an acid when adsorbing the phenylenediamine compound, electrode modifier, electrode adsorbent, or electron transfer enhancer of the present invention to the electrode. In another embodiment, the electrode may be pretreated with an acid before the phenylenediamine compound, electrode modifier, electrode adsorbent, or electron transfer promoter of the present invention is adsorbed onto the electrode. Note that the pretreatment with an acid herein includes bringing an electrolyte solution containing an acid into contact with the electrode. Further, in this specification, acid refers to an acid having a pH of 4 or less, for example, less than pH 4, less than pH 3, less than pH 3, less than pH 2, less than pH 2, or pH 1. In one embodiment, the phenylenediamine compound, electrode modifier, electrode adsorbent, or electron transfer promoter of the present invention is converted into an oxidized or reduced form by applying a specific potential, and then adsorbed onto the electrode. Also good. Note that an oxidizing agent or a reducing agent may be used as a method for converting into an oxidized form or a reduced form, and the method is not particularly limited. In one embodiment, the phenylenediamine compound, electrode modifier, electrode adsorbent, or electron transfer promoter of the present invention may be incorporated into a polymer and adsorbed onto the electrode. Such adsorption may be referred to herein as embedded or entrapped. This is distinguished from adsorption fixation, that is, fixation by physical adsorption. In inclusion fixation or entrapping fixation, the compound of the invention can diffuse in the polymer, whereas in adsorption fixation, the compound of the invention does not or hardly diffuses. In another embodiment, the compound, electrode modifier, electrode adsorbent, or electron transfer enhancer of the present invention is applied to an ionic polymer, such as a cationic polymer such as polyethyleneimine or polylysine, an anionic polymer such as polyaniline, or polyacrylic acid. It may be adsorbed to the electrode by electrostatic interaction using, for example. In another embodiment, the compound of the present invention, electrode modifying agent, electrode adsorbent, or electron transfer enhancer may be adsorbed onto an enzyme or immobilized using a crosslinking agent, and may be immobilized on the electrode together with the enzyme. In another embodiment, the adsorbed compounds of the invention may be polymerized by redox reactions or by crosslinking agents.

［式中、
R¹は、-NR⁷R⁸、-N=N-R⁹、又は-N⁺≡Nであり、
R²は、-NR¹⁰R¹¹又は、-N=N-R¹²であり、
R⁷及びR⁸は、それぞれ独立に、水素、場合により1以上のX又はVにより置換されてもよい、直鎖又は分枝鎖のC_1-7アルキル、C_1-7アルケニル、C_1-7アルキニル、C_3-9シクロアルキル、フェニル、1-ナフチル、2-ナフチル、アントラセニル、フェナントレニル、アセチル、カルボキシ、フラニルホルミル、ピラゾリルホルミル、1-メチル-1H-ピラゾール-5-イルホルミル、9,9-ジメチルフルオレン-2-イル、-N⁺≡N、-N=N-フェニル、ベンジル、

であり、
R¹⁰は、水素、場合により1以上のX又はVにより置換されてもよい、直鎖又は分枝鎖のC_1-7アルキル、C_1-7アルケニル、C_1-7アルキニル、C_3-9シクロアルキル、フェニル、1-ナフチル、2-ナフチル、アントラセニル又はフェナントレニル、であり、
R³、R⁴、R⁵及びR⁶は、それぞれ独立に、水素、場合により1以上のYにより置換されてもよい、直鎖又は分枝鎖のC_1-7アルキル、C_1-7アルケニル、C_1-7アルキニル、C_1-7アルコキシ、ハロ、ニトロ、シアノ、カルボキシ、スルホ、ヒドロキシ又はアミノであり、
或いは、R³及びR⁴、又は、R⁵及びR⁶は、それらを含有するベンゼン環と一緒になって、場合により1以上のオキソ、X又はWにより置換されてもよい、ベンゼン環、又は

を形成し、但し＊はR³が結合する炭素原子に結合し、＊＊はR⁴が結合する炭素原子に結合するか、又は、＊はR⁵が結合する炭素原子に結合し、＊＊はR⁶が結合する炭素原子に結合し、
R¹¹は、場合により１以上のX又はZにより置換されてもよい、フェニル、1-ナフチル、2-ナフチル、アントラセニル及びフェナントレニルからなる群より選択され、
R⁹は、水素、場合により１以上のXにより置換されてもよい、フェニル、1-ナフチル、2-ナフチル、アントラセニル及びフェナントレニルからなる群より選択され、
R¹²は、場合により１以上のX、W、イソチオシアネート、又はハロスルホニルにより置換されてもよい、フェニル、1-ナフチル、2-ナフチル、アントラセニル、フェナントレニル、及び

からなる群より選択され、
ここでVは、場合によりC1-7アルキルに置換されてもよい、-O-アクリロイル、アセチルアミノ、又はフェニルであり、
Wは、場合により1以上のアミノ、C1-7アルキル、アミノアルキルにより置換されてもよい、D若しくはL-アラニルスルホニル、D若しくはL-バリルスルホニル、D若しくはL-ロイシルスルホニル、D若しくはL-メチオニルスルホニル、D若しくはL-プロリルスルホニル、D若しくはL-トリプトフィルスルホニル、D若しくはL-グリシルスルホニル、D若しくはL-システイニルスルホニル、D若しくはL-イソロイシルスルホニル、D若しくはL-フェニルアラニルスルホニル、D若しくはL-チロシルスルホニル、D若しくはL-セリルスルホニル、D若しくはL-トレオニルスルホニル、D若しくはL-アスパラギニルスルホニル、D若しくはL-グルタミルスルホニル、D若しくはL-アルギニルスルホニル、D若しくはL-ヒスチジルスルホニル、D若しくはL-リジルスルホニル、D若しくはL-アスパラギルスルホニル、D若しくはL-グルタミニルスルホニル、-C(=O)-O-スクシンイミジル、アセチル、トリフルオロアセチル、ベンゾイルアミノ、-N=N-フェニル、フェニルアミノ、ジアミノフェニルアゾフェニル、又は、場合により１以上のXにより置換されてよいフェニルアゾ、場合により１以上のYにより置換されてもよいナフチルアゾ、アセチルアミノ、

であり、
Xは、場合により1以上のハロ、アミノ、シアノ、カルボキシ、カルボニル、アルコキシ、アルキルアミノ、ニトロソ、ニトロ及びスルホからなる群より選択される置換基により置換されてもよい、直鎖又は分枝鎖の、C_1-7アルキル、C_1-7アルケニル、C_1-7アルキニル、C_1-7アルコキシ、ハロ、ヒドロキシ、ニトロ、カルボキシ、シアノ、スルホ、アミノ又はアルキルアミノであり、
Yはハロ、アミノ、シアノ、カルボキシ、カルボニル、ヒドロキシ、アルコキシ及びスルホからなる群より選択され、
Zは-SO₂-CH=CH₂、又は-SO₂-C₂H₄-O-SO₃H、4,6-ジクロロトリアジン-2-イルアミノである］。 In certain embodiments, the phenylenediamine-based compound of the present invention can be a compound of general formula I or II below, or a salt, anhydride, or solvate thereof:

[In the formula,
R ¹ is -NR ⁷ R ⁸ , -N=NR ⁹ , or -N ⁺ ≡N,
R ² is -NR ¹⁰ R ¹¹ or -N=NR ¹² ,
R ⁷ and R ⁸ are each independently hydrogen, a straight chain or branched C _1-7 alkyl, C _1-7 alkenyl, C _1- which may optionally be substituted with one or more X or V; ₇ alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, acetyl, carboxy, furanylformyl, pyrazolylformyl, 1-methyl-1H-pyrazol-5-ylformyl, 9,9 -dimethylfluoren-2-yl, -N ⁺ ≡N, -N=N-phenyl, benzyl,

and
R ¹⁰ is hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _3-9 optionally substituted by one or more X or V; cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl,
R ³ , R ⁴ , R ⁵ and R ⁶ are each independently hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl, optionally substituted with one or more Y; , C _1-7 alkynyl, C _1-7 alkoxy, halo, nitro, cyano, carboxy, sulfo, hydroxy or amino,
Alternatively, R ³ and R ⁴ or R ⁵ and R ⁶ together with the benzene ring containing them are optionally substituted with one or more oxo, X or W, or

, where * is bonded to the carbon atom to which R ³ is bonded, ** is bonded to the carbon atom to which R ⁴ is bonded, or * is bonded to the carbon atom to which R 5 is bonded, and ** is bonded to the carbon atom to which R ⁵ is bonded. is bonded to the carbon atom to which R ⁶ is bonded,
R ¹¹ is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X or Z;
R ⁹ is selected from the group consisting of hydrogen, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X;
R ¹² is phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, optionally substituted with one or more of X, W, isothiocyanate, or halosulfonyl;

selected from the group consisting of
Here, V is -O-acryloyl, acetylamino, or phenyl, which may be optionally substituted with C1-7 alkyl,
W is D or L-alanylsulfonyl, D or L-valylsulfonyl, D or L-leucylsulfonyl, D or L, optionally substituted with one or more amino, C1-7 alkyl, aminoalkyl -methionylsulfonyl, D or L-prolylsulfonyl, D or L-tryptophylsulfonyl, D or L-glycylsulfonyl, D or L-cysteinylsulfonyl, D or L-isoleuylsulfonyl, D or L-phenylalanylsulfonyl, D or L-tyrosylsulfonyl, D or L-serylsulfonyl, D or L-threonylsulfonyl, D or L-asparaginylsulfonyl, D or L-glutamylsulfonyl, D or L- Arginylsulfonyl, D or L-histidylsulfonyl, D or L-lysylsulfonyl, D or L-asparagylsulfonyl, D or L-glutaminylsulfonyl, -C(=O)-O-succinimidyl, acetyl, tri- fluoroacetyl, benzoylamino, -N=N-phenyl, phenylamino, diaminophenylazophenyl, or phenylazo optionally substituted by one or more X, naphthylazo optionally substituted by one or more Y, acetylamino,

and
X is a straight or branched chain, optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy, alkylamino, nitroso, nitro, and sulfo. is C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _1-7 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo, amino or alkylamino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, hydroxy, alkoxy and sulfo;
Z is _-SO2 -CH= _CH2 , or _-SO2 - _C2H4 - _O - _SO3H , 4,6-dichlorotriazin-2-ylamino].

［式中、
R^1aは、-NR^7aR^8a、-N=N-R^9a、又は-N⁺≡Nであり、
R^2aは、-NR^10aR^11a、又は-N=N-R^12aであり、
R^7a及びR^8aは、それぞれ独立に、水素、場合により1以上のXaにより置換されてもよい、直鎖又は分枝鎖のC_1-6アルキル、C_1-6アルケニル、C_1-6アルキニル、C_3-9シクロアルキル、フェニル、1-ナフチル、2-ナフチル、アントラセニル、又はフェナントレニルであり、
R¹⁰は、水素、場合により1以上のXaにより置換されてもよい、直鎖又は分枝鎖のC_1-6アルキル、C_1-6アルケニル、C_1-6アルキニル、C_3-9シクロアルキル、フェニル、1-ナフチル、2-ナフチル、アントラセニル又はフェナントレニルであり、
R^3a、R^4a、R^5a及びR^6aは、それぞれ独立に、水素、場合により1以上のYにより置換されてもよい、直鎖又は分枝鎖のC_1-6アルキル、C_1-6アルケニル、C_1-6アルキニル、C_1-6アルコキシ、ハロ、ニトロ、シアノ、カルボキシ、スルホ、ヒドロキシ又はアミノであり、
或いは、R^3a及びR^4a、又は、R^5a及びR^6aは、ベンゼン環、又は

を形成し、但し＊はR^3aが結合する炭素原子に結合し、＊＊はR^4aが結合する炭素原子に結合するか、又は、＊はR^5aが結合する炭素原子に結合し、＊＊はR^6aが結合する炭素原子に結合し、
R^11aは、場合により１以上のXaにより置換されてもよい、フェニル、1-ナフチル、2-ナフチル、アントラセニル及びフェナントレニル、からなる群より選択され、
R^9aおよびR^12aは、それぞれ独立に、水素、場合により１以上のXaにより置換されてもよい、フェニル、1-ナフチル、2-ナフチル、アントラセニル及びフェナントレニル、からなる群より選択され、
Xaは、場合により1以上のハロ、アミノ、シアノ、カルボキシ、カルボニル、ヒドロキシ、アルコキシ、アルキルアミノ、ニトロソ、ニトロ及びスルホからなる群より選択される置換基により置換されてもよい、直鎖又は分枝鎖の、C_1-6アルキル、C_1-6アルケニル、C_1-6アルキニル、C_1-6アルコキシ、ハロ、ヒドロキシ、ニトロ、カルボキシ、シアノ、スルホ、アミノ又はアルキルアミノであり、
Yはハロ、アミノ、シアノ、カルボキシ、カルボニル、アルコキシ及びスルホからなる群より選択される］。 In some embodiments, the phenylenediamine-based compound of the present invention may be a compound having the structure of general formula Ia or IIa below, or a salt, anhydride, or solvate thereof:

[In the formula,
R ^1a is -NR ^7a R ^8a , -N=NR ^9a , or -N ⁺ ≡N,
R ^2a is -NR ^10a R ^11a or -N=NR ^12a ,
R ^7a and R ^8a are each independently hydrogen, linear or branched C _1-6 alkyl, C _1-6 alkenyl, C _1-6 alkynyl, optionally substituted with one or more Xa; , C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, or phenanthrenyl,
R ¹⁰ is hydrogen, linear or branched C _1-6 alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _3-9 cycloalkyl, optionally substituted with one or more Xa , phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl,
R ^3a , R ^4a , R ^5a and R ^6a are each independently hydrogen, linear or branched C _1-6 alkyl, C _1-6 alkenyl, optionally substituted with one or more Y; , C _1-6 alkynyl, C _1-6 alkoxy, halo, nitro, cyano, carboxy, sulfo, hydroxy or amino,
Alternatively, R ^3a and R ^4a or R ^5a and R ^6a are a benzene ring, or

, where * is bonded to the carbon atom to which R ^3a is bonded, ** is bonded to the carbon atom to which R ^4a is bonded, or * is bonded to the carbon atom to which R ^{5a is bonded, and ** is bonded to the carbon atom to which R 5a} is bonded. is bonded to the carbon atom to which R ^6a is bonded,
R ^11a is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more Xa;
R ^9a and R ^12a are each independently selected from the group consisting of hydrogen, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted with one or more Xa;
Xa is a linear or branched chain, optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, hydroxy, alkoxy, alkylamino, nitroso, nitro and sulfo. branched C _1-6 alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _1-6 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo, amino or alkylamino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo].

［式中、
R^7b、R^8b及びR^10bは、それぞれ独立に、水素、場合により1以上のXbにより置換されてもよい、直鎖又は分枝鎖のC_1-6アルキル、C_1-6アルケニル、C_1-6アルキニル、C_3-9シクロアルキル、フェニル、1-ナフチル、2-ナフチル、アントラセニル又はフェナントレニル、であり、R^3b、R^4b、R^5b及びR^6bは、それぞれ独立に、水素、場合により1以上のYにより置換されてもよい、直鎖又は分枝鎖のC_1-6アルキル、C_1-6アルケニル、C_1-6アルキニル、C_1-6アルコキシ、ハロ、ニトロ、シアノ、カルボキシ、スルホ又はアミノであり、
R^11bは、場合により１以上のXbにより置換されてもよい、フェニル、1-ナフチル、2-ナフチル、アントラセニル及びフェナントレニル、からなる群より選択され、
ここでXbは、場合により1以上のハロ、アミノ、シアノ、カルボキシ、カルボニル、アルコキシ及びスルホからなる群より選択される置換基により置換されてもよい、直鎖又は分枝鎖の、C_1-6アルキル、C_1-6アルケニル、C_1-6アルキニル、C_1-6アルコキシ、ハロ、ヒドロキシ、ニトロ、カルボキシ、シアノ、スルホ又はアミノであり、
Yはハロ、アミノ、シアノ、カルボキシ、カルボニル、アルコキシ及びスルホからなる群より選択される］。別の実施形態では、上記一般式Ｉ若しくはIIで表される本発明の化合物又は上記一般式Ｉa若しくはIIaで表される本発明の化合物から、上記一般式Ib若しくはIIｂの構造を有する化合物、その塩若しくは溶媒和物の一部又は全部は除かれる。 In some embodiments, the phenylenediamine-based compound of the present invention may be a compound having the structure of general formula Ib or IIb below, or a salt, anhydride, or solvate thereof:

[In the formula,
R ^7b , R ^8b and R ^10b are each independently hydrogen, a linear or branched C _1-6 alkyl, C _1-6 alkenyl, C ₁ which may optionally be substituted with one or more Xb _-6 alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl, and R ^3b , R ^4b , R ^5b and R ^6b are each independently hydrogen, optionally 1 Straight chain or branched C _1-6 alkyl, C _1-6 alkenyl, C _{1-6 alkynyl, C 1-6 alkoxy} , halo, nitro, cyano, carboxy, sulfo, which may be substituted by the above Y or amino;
R ^11b is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more Xb;
Here, Xb is a linear or branched C 1- which may be optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo _{. 6} alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _1-6 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo or amino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo]. In another embodiment, from the compound of the present invention represented by the above general formula I or II or the compound of the present invention represented by the above general formula Ia or IIa, a compound having the structure of the above general formula Ib or IIb, Some or all salts or solvates are excluded.

Alkyl, as used herein, refers to a straight-chain or branched hydrocarbon having, for example, 1 to 7 carbon atoms, such as 1 to 6 carbon atoms. Examples of alkyl include, but are not limited to, methyl, ethyl, propyl, isopropyl, isobutyl, n-butyl, tert-butyl, isopentyl, n-pentyl, heptyl.

As used herein, the number of atoms (such as carbon atoms) is expressed, for example, as "Cx-Cy alkyl," which refers to an alkyl group having x to y carbon atoms. Similar notations are used for other substituents and ranges.

As used herein, alkenyl refers to a straight or branched chain aliphatic hydrocarbon having one or more carbon-carbon double bonds. Examples include, but are not limited to, vinyl, allyl, and the like.

As used herein, alkynyl refers to a straight or branched chain aliphatic hydrocarbon having one or more carbon-carbon triple bonds. Examples include, but are not limited to, ethynyl.

Cycloalkyl, as used herein, refers to a substituted or unsubstituted non-aromatic cyclic hydrocarbon ring. Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

As used herein, halo refers to a Group 17 element group, such as -Cl, -Br, -I, and the like. As used herein, halogen refers to fluorine, chlorine, bromine, or iodine.

As used herein, haloalkyl refers to an alkyl group substituted with at least one halogen. Haloalkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, and t- independently substituted with one or more halogens, such as fluoro, chloro, bromo, and iodo. Examples include butyl.

Halosulfonyl as used herein refers to a sulfonyl group substituted with at least one halogen, such as chlorosulfonyl (Cl-SO ₂ -), bromosulfonyl (Br-SO ₂ -), etc. Not limited to.

As used herein, phenyl refers to substituted or unsubstituted benzene ring systems. Naphthyl, as used herein, refers to substituted or unsubstituted naphthalene ring systems and includes 1-naphthyl and 2-naphthyl. As used herein, anthracenyl refers to a substituted or unsubstituted anthracene ring system. As used herein, phenanthrenyl refers to substituted or unsubstituted phenanthrene ring systems.

Acetyl as used herein refers to CH ₃ CO-. As used herein, trifluoroacetyl refers to CF ₃ CO-. Carboxy as used herein refers to -COOH. As used herein, furanyl refers to a monovalent group of furan, such as 2-furanyl or 3-furanyl. In this specification, formyl is also referred to as aldehyde and refers to -COH. In this specification, furanylformyl refers to a formyl group (furanyl-CO-) to which a furanyl group is bonded. As used herein, pyrazolyl refers to a pyrazole ring system. In this specification, pyrazolylformyl refers to a formyl group to which a pyrazolyl group is bonded (pyrazolyl-CO-). Fluorenyl, as used herein, refers to substituted or unsubstituted fluorene ring systems.

In this specification, -N ⁺ ≡N is also referred to as azide, and is also expressed as -N ₂ group. As used herein, benzyl refers to C ₆ H ₅ CH ₂ -. As used herein, benzoyl refers to C ₆ H ₅ -C(=O)-. In this specification, azo is also expressed as R'-N=N-R'', and R' and R'' may be the same or different. For example, naphthylazo represents naphthyl-N=N-.

As used herein, nitro refers to the _-NO2 group. As used herein, nitroso refers to the group -N=O. As used herein, cyano refers to the -CN group. As used herein, sulfo refers to -SO ₃ H. As used herein, hydroxy refers to -OH. As used herein, oxo refers to =O.

In this specification, amino refers to a -NR'R'' group, and R' and R'' may be the same or different. R' and R'' can be, for example, but not limited to, H, alkyl, alkenyl, alkynyl, cycloalkyl, phenyl, naphthyl, anthracenyl, phenanthrenyl, and the like. As used herein, aminoalkyl has an alkylene linker to which an amino group is attached. Examples of aminoalkyl include, but are not limited to, -(CH ₂ ) _n NH ₂ and the like. In this specification, alkylamino refers to amino to which an alkyl group is linked. Examples of alkylamino include, but are not limited to, C _1-7 alkyl-NH-. In this specification, acetylamino refers to amino to which an acetyl group is linked. Examples of acetylamino include, but are not limited to, acetyl-NH-.

In this specification, acryloyl refers to H ₂ C=CH-C(=O)-. In this specification, -O-acryloyl refers to H ₂ C=CH-C(=O)-O-. In this specification, isothiocyanate refers to -N=C=S. As used herein, succinimidyl refers to (CH ₂ CO) ₂ N-. In this specification, carbonyl refers to -C(=O)-. As used herein, alkoxy refers to an -O-alkyl group.

As used herein, "optionally substituted" and "substituted or unsubstituted" mean any substitution with one or more substituents, and this also includes multiple substitutions. It will be done.

であり得る（CAS No. 101-72-4）。別の実施形態では、本発明の化合物から、N-イソプロピル-N'-フェニル-p-フェニレンジアミン（IPPD）は除かれる。 In certain embodiments, the phenylenediamine-based compound of the invention is N-isopropyl-N'-phenyl-p-phenylenediamine (IPPD).

(CAS No. 101-72-4). In another embodiment, the compounds of the invention exclude N-isopropyl-N'-phenyl-p-phenylenediamine (IPPD).

であり得る（Cas No. 74-31-7）。別の実施形態では、本発明の化合物から、N,N'-ジフェニル-p-フェニレンジアミン（DPPD）は除かれる。 In certain embodiments, the phenylenediamine-based compound of the invention is N,N'-diphenyl-p-phenylenediamine (DPPD).

(Cas No. 74-31-7). In another embodiment, the compounds of the invention exclude N,N'-diphenyl-p-phenylenediamine (DPPD).

であり得る（CAS No. 793-24-8）。別の実施形態では、本発明の化合物から、N-(1,3-ジメチルブチル)-N'-フェニル-p-フェニレンジアミン（6PPD）は除かれる。 In certain embodiments, the phenylenediamine-based compound of the present invention is N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD).

(CAS No. 793-24-8). In another embodiment, the compounds of the invention exclude N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD).

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is Bindschaedler's Green Leuco Base (BGLB, CAS No. 637-31-0).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is Variamine Blue B Base (CAS No. 101-64-4).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is 2-nitro-4-aminodiphenylamine (CAS No. 2784-89-6).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the present invention is N-methyl-N'-phenyl-p-phenylenediamine.

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the present invention is N-ethyl-N'-phenyl-p-phenylenediamine.

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the present invention is N-isopropyl-N'-(4-aminophenyl)-p-phenylenediamine.

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is tris[4-(diethylamino)phenyl]amine (TDPA, CAS No. 47743-70-4).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the present invention is 4-(dimethylamino)-4'-nitrosodiphenylamine (CAS No. 7696-70-0).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is N-phenyl-o-phenylenediamine (CAS No. 534-85-0).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the present invention is N-(4-chlorophenyl)-1,2-phenylenediamine (CAS No. 68817-71-0).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is 4-diazodiphenylamine sulfate (CAS No. 4477-28-5).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the present invention is Acid Yellow 36 (CAS number 587-98-4).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is 2-amino-4-isopropylamino-diphenylamine.

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the present invention is N-isopropyl-N'-(4-hydroxyphenyl)-p-phenylenediamine.

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the present invention is N,N'-di-2-naphthyl-1,4-phenylenediamine (CAS No. 93-46-9).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is 4-(2-octylamino)diphenylamine (CAS No. 15233-47-3).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is N-(2-Amino-4-chlorophenyl)anthranilic acid (CAS No. 67990-66-3).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is 4-diazo-3-methoxydiphenylamine sulfate (CAS No. 36305-05-2).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is 4-(phenylazo)diphenylamine (CAS No. 101-75-7).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is Acid Orange 5 (CAS RN: 554-73-4).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is Alizarin Cyanin Green F (CAS No. 4403-90-1).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is Alizarin Astrol (CAS RN: 6408-51-1)

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is Disperse Yellow 9 (CAS No. 6373-73-5).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the present invention is 5-Sulfo-4'-diethylamino-2,2'-dihydroxyazobenzene (CAS No. 1563-01-5).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the present invention is Alizarin Cyanin Green F (CAS number 4403-90-1).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the present invention is Alphamine Red R Base (CAS number 57322-42-6).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is Crocein scarlet 3B (CAS number 5413-75-2).

It can be.

であり得る。 In certain embodiments, the phenylenediamine-based compound of the invention is p-phenylphenylenediamine (CAS No. 2198-59-6).

It can be.

In certain embodiments, the phenylenediamine-based compounds of the invention can be the compounds in the table below.

を有するp-フェニレンジアミン及びo-フェニレンジアミンは除かれる。また、本発明のフェニレンジアミン系化合物からN,N-ジメチル-p-フェニレンジアミン、N,N-ジメチル-o-フェニレンジアミン及びN,N,N',N'-テトラメチルフェニレンジアミンは除かれる。 From the phenylenediamine compound of the present invention, the following structure

Excludes p-phenylenediamine and o-phenylenediamine with Furthermore, N,N-dimethyl-p-phenylenediamine, N,N-dimethyl-o-phenylenediamine and N,N,N',N'-tetramethylphenylenediamine are excluded from the phenylenediamine compounds of the present invention.

（式中、Ｘ₁～Ｘ₉は、それぞれ独立して、水素、弗素、塩素、臭素、シアノ基、ニトロ基、－Ｎ(Ｒ1)2、鎖状飽和炭化水素、鎖状不飽和炭化水素、環状飽和炭化水素、または環状不飽和炭化水素を表し、Ｒ1は、水素、鎖状飽和炭化水素、鎖状不飽和炭化水素、環状飽和炭化水素、環状不飽和炭化水素、シアノ基、ニトロ基およびそれらの組み合わせからなる群より選ばれる少なくとも１種である。ただし、Ｘ1～Ｘ9がすべて水素である場合を除く。）。別の実施形態では、電池に用いる本発明の化合物から、該構造を有するトリフェニルアミン誘導体は除かれる。 In certain embodiments, the compounds of the invention exclude triphenylamine derivatives having the following structure of general formula (1):

(In the formula, X ₁ to X ₉ are each independently hydrogen, fluorine, chlorine, bromine, cyano group, nitro group, -N(R1)2, chain saturated hydrocarbon, chain unsaturated hydrocarbon, Represents a cyclic saturated hydrocarbon or a cyclic unsaturated hydrocarbon, and R1 represents hydrogen, a chain saturated hydrocarbon, a chain unsaturated hydrocarbon, a cyclic saturated hydrocarbon, a cyclic unsaturated hydrocarbon, a cyano group, a nitro group, and the like. At least one type selected from the group consisting of combinations of (However, this excludes cases where all of X1 to X9 are hydrogen.) In another embodiment, triphenylamine derivatives having this structure are excluded from the compounds of the invention for use in batteries.

In another embodiment, the compounds of the invention exclude N,N'-diphenyl-N,N'-bis(p-tolyl-1,4-phenylenediamine). The compounds of the invention exclude N,N'-diphenyl-N,N'-bis(p-tolyl-1,4-phenylenediamine).

The phenylenediamine compound of the present invention may be modified with a functional group for covalently bonding it to an enzyme. A C1 to C20 alkyl, amino acid, peptide, etc. may be inserted as a linker between the phenylenediamine compound and the functional group. A hydroxyl group, an amino group, an alkene, etc. may be included between the linkers as appropriate. Examples of functional groups include hydroxyl group, carboxyl group, amino group, aldehyde group, hydrazino group, thiocyanate group, epoxy group, vinyl group, halogen group, acid ester group, phosphoric acid group, thiol group, disulfide group, dithiocarbamate group. , dithiophosphate group, dithiophosphonate group, thioether group, thiosulfate group, succinimide group, maleimide group, and thiourea group.

A phenylenediamine compound can be in a redox state and an ionized state. In the above chemical formula, the phenylenediamine compound of the present invention is described in a neutral and reduced form. However, in addition to this form, the phenylenediamine compound of the present invention may be in an oxidized type (diimine type), a semi-oxidized type, or a reduced type (diamine type). The azo compounds or diazonio compounds of the present invention may similarly be in the oxidized, semi-oxidized, or reduced form. Further, the phenylenediamine compound of the present invention may be a neutral type or a cationic type. For convenience, when referring to the phenylenediamine compound of the present invention, for example, the phenylenediamine compound of the present invention represented by the above chemical formula, this refers to a neutral type, a cationic type, an oxidized type, a half-oxidized type, or a reduced type. It shall include things of the form. For example, after adding a neutral oxidized compound as the phenylenediamine compound of the present invention to the measurement system, it may change to an oxidized cationic compound due to the pH of the solution or electron transfer. However, such compounds are also included in the phenylenediamine compounds of the present invention. Furthermore, when referring to the phenylenediamine compound of the present invention, this includes its salts, acid addition salts, anhydrides, and solvates. Examples of the salt include, but are not limited to, salts of Group 1 elements and salts of Group 17 elements, such as Na salts, K salts, Cl salts, and Br salts. Acid addition salts include, but are not limited to, hydrochlorides, sulfates, sulfites, and nitrates.

The phenylenediamine compound of the present invention may be artificially synthesized or may be obtained from a natural product. It may also be a commercially available one. In the case of synthesis, organic synthesis can be performed using conventional organic synthesis techniques, and the product can be confirmed by NMR, IR, mass spectrometry, etc. The practice of the present invention will employ, unless otherwise indicated, conventional techniques of chemistry, organic synthesis, biochemistry, molecular biology, and electrochemistry, which are within the ability of those skilled in the art. Such techniques are explained in the literature. For example, Organic Chemistry (Jonathan Clayden (editor), Nick Greeves (author), Stuart Warren (author), Peter Wothers (author)), Oxford Univ Pr, 2000, and March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure (Michael B. Smith (author), Jerry March (author)) Wiley-Interscience, 6th edition, 2007. Each of these general texts is incorporated herein by reference.

(Method for immobilizing enzyme or oxidoreductase)
Enzymes or oxidoreductases can be immobilized on a solid phase by any known method. Enzymes, such as oxidoreductases, may be immobilized on beads, membranes, carbon particles, gold particles, platinum particles, polymers, and electrode surfaces. Immobilization methods include methods using crosslinking reagents, encapsulation in polymer matrices, coating with dialysis membranes, photocrosslinkable polymers, conductive polymers, and redox polymers. It may be fixed by suction on the surface, or a combination of these may be used. Typically, after a redox enzyme is immobilized on a carbon electrode using glutaraldehyde, the glutaraldehyde is blocked by treatment with a reagent having an amine group. The amount of the oxidoreductase to be immobilized can be determined as appropriate so as to generate the current necessary for electrochemical measurement or fuel cell power generation.

(Method for adsorbing phenylenediamine compounds of the present invention)
In some embodiments, the phenylenediamine compound of the present invention may exist in a free state in a solution, or may be adsorbed on beads, membranes, carbon particles, gold particles, platinum particles, polymers, or electrode surfaces. For example, it may be physically adsorbed. The fact that the phenylenediamine compound of the present invention is adsorbed on the electrode surface may also be referred to as being immobilized on the electrode surface, but in this specification, these terms are synonymous. Examples of the adsorption method include a method including a step of dissolving the phenylenediamine compound of the present invention in a suitable medium and bringing the solution into physical contact with an electrode. In another embodiment, the phenylenediamine-based compounds of the present invention may be sprayed onto the electrode. The amount of the phenylenediamine compound to be adsorbed can be determined as appropriate so as to generate the current necessary for electrochemical measurement or battery power generation.

In one embodiment, the phenylenediamine compound of the present invention may be adsorbed on the electrode, and an enzyme, such as an oxidoreductase, may also be immobilized thereon. In this case, the phenylenediamine compound of the present invention may be adsorbed first, and then an enzyme such as an oxidoreductase may be immobilized, or the enzyme such as an oxidoreductase may be first immobilized and then the phenylenediamine compound of the present invention is adsorbed. Alternatively, a phenylenediamine-based compound may be adsorbed at the same time in the process of immobilizing an enzyme, such as an oxidoreductase.

The final concentration of the phenylenediamine compound of the present invention added to the sample solution is not particularly limited, and for example, 1 pM or more, 2 pM or more, 3 pM or more, 4 pM or more, 5 pM or more, 6 pM or more, 7 pM or more, 8 pM or more, 9 pM or more, 10pM or more, 1M or less, 100mM or less, 20mM or less, 10mM or less, 5mM or less, 1mM or less, 800μM or less, 600μM or less, 500μM or less, 400μM or less, 300μM or less, 200μM or less, 100μM or less, 50μM or less, such as 1pM to 1M, 1pM to 100mM, 1pM to 20mM, 1pM to 10mM, 1pM to 5mM, 2pM to 1mM, 3pM to 800μM, 4pM to 600μM, 5pM to 500μM, 6pM to 400μM, 7pM to 300μM, 8pM to 200μM, 9pM to 100μM, 10pM to It can be in the range of 50 μM. The final concentration of the phenylenediamine compound of the present invention added to the sample solution is not particularly limited, and is, for example, 0.000001 to 0.5% (w/v), 0.000003 to 0.3% (w/v), or 0.000005 to 0.1% (w/v). ), 0.00001-0.05% (w/v), 0.00002-0.03% (w/v), 0.00003-0.01% (w/v). Note that the order of addition of the phenylenediamine compound and other reagents is not limited, and may be added simultaneously or sequentially.

In certain embodiments, the time for performing the redox reaction or the time for performing the electrochemical measurement can be 60 minutes or less, 30 minutes or less, 10 minutes or less, 5 minutes or less, or 1 minute or less. Alternatively, for enzyme sensors, batteries, etc. that are used for long-term measurements, the redox reaction time is 60 minutes or more, 120 minutes or more, 1 day or more, 2 days or more, 3 days or more, 1 week or more, 2 weeks or more, It can be for more than 3 weeks. For example, when a phenylenediamine compound is used together with an oxidoreductase, the concentration of each component of the electrochemical measurement reagent is determined in accordance with the concentration range of the reduced mediator contained in the sample or estimated to be generated in the sample. can be adjusted.

Unless otherwise specified, the enzyme contained in the composition of the present invention or the enzyme immobilized on the electrode of the present invention is a purified enzyme. Cell extracts, cell disruption solutions, and crude enzyme extracts containing enzymes contain various contaminants in addition to enzymes. For example, it has been reported that the amount of riboflavin in a crude enzyme extract of microorganisms is approximately 53 to 133 μM (J Indust Micro Biotech 1999, 22, pp. 8-18). If such a crude enzyme extract is subjected to electrochemical measurement as it is, contaminants will receive electrons and interfere with the transfer of electrons to the electrode. Accurate electrochemical measurements are difficult. Therefore, in the electrochemical measurement method of the present invention, enzymes from which contaminants have been removed can be used. In this specification, the term "enzyme is purified" or "purified enzyme" does not necessarily mean that the protein is pure, but that contaminants have been removed from the enzyme preparation to the extent that electrochemical measurement is possible. It means to be there.

(Measurement of oxidoreductase activity)
In explaining the measurement of oxidoreductase activity, glucose dehydrogenase (GDH) will be used as an example of a specific enzyme. GDH (EC 1.1.99.10) catalyzes the reaction that oxidizes the hydroxyl group of glucose to produce glucono-δ-lactone. At this time, the electron acceptor receives electrons and becomes a reduced electron acceptor. The activity of GDH can be measured using this principle of action, for example, using the following measurement system using phenazine methosulfate (PMS) and 2,6-dichloroindophenol (DCIP) as electron acceptors. .
(Reaction 1) D-glucose + PMS (oxidized type)
→ D-glucono-δ-lactone + PMS (reduced form)
(Reaction 2) PMS (reduced type) + DCIP (oxidized type)
→ PMS (oxidized type) + DCIP (reduced type)

Specifically, first, in (reaction 1), PMS (reduced type) is generated as D-glucose is oxidized. In the subsequent reaction (reaction 2), DCIP is reduced as PMS (reduced form) is oxidized. The degree of disappearance of this "DCIP (oxidized form)" is detected as the amount of change in absorbance at a wavelength of 600 nm, and the enzyme activity can be determined based on this amount of change.

GDH activity can be measured according to the following procedure. Mix 2.05mL of 100mM phosphate buffer (pH 7.0), 0.6mL of 1M D-glucose solution, and 0.15mL of 2mM DCIP solution, and incubate at 37°C for 5 minutes. Next, 0.1 mL of 15 mM PMS solution and 0.1 mL of enzyme sample solution are added to start the reaction. The absorbance at the start of the reaction and over time is measured, the amount of decrease in absorbance at 600 nm per minute (ΔA600) as the enzyme reaction progresses is determined, and the GDH activity is calculated according to the following formula. In this case, GDH activity is defined as 1 U, which is the amount of enzyme that reduces 1 μmol of DCIP per minute in the presence of D-glucose at a concentration of 200 mM at 37°C.

In addition, 3.0 in the formula is the volume of reaction reagent + enzyme reagent (mL), 16.3 is the millimolar molecular extinction coefficient (cm ² /μmol) under this activity measurement condition, 0.1 is the volume of enzyme solution (mL), and 1.0 is the volume of enzyme solution (mL). The optical path length of the cell (cm), ΔA600 _blank is the amount of decrease in absorbance per minute at 600 nm when the reaction is started by adding 100mM phosphate buffer (pH 7.0) instead of the enzyme sample solution, and df is the dilution. Represents a multiple.

The electrochemical measurement kit of the present invention contains the phenylenediamine compound of the present invention or an electrode modifier containing the phenylenediamine compound in an amount sufficient for at least one assay. Typically, the electrochemical measurement of the present invention includes, in addition to the phenylenediamine compound of the present invention, an oxidoreductase, a buffer necessary for the assay, a substrate standard solution for preparing a calibration curve, and guidelines. including. For example, the oxidoreductase may be GDH, in which case the substrate standard solution may be a glucose standard solution.

In one embodiment, the electrochemical measurement kit of the present invention contains the phenylenediamine compound of the present invention and an oxidoreductase as the same reagent. In another embodiment, the electrochemical measurement kit of the present invention contains a phenylenediamine compound and an oxidoreductase as separate reagents. In another embodiment, the oxidoreductase may be immobilized on an electrode, and the electrochemical measurement kit of the present invention used for such an electrode contains a phenylenediamine-based compound as a single reagent. However, the term "single reagent" as used herein does not mean that the reagent does not contain substances other than the phenylenediamine compound. The single reagent may contain a suitable medium so that the phenylenediamine-based compound of the present invention is dissolved in the single reagent. The medium may be any medium in which the phenylenediamine compound of the present invention can be dissolved, and examples include, but are not limited to, water, methanol, ethanol, propanol, acetone, and acetonitrile. The phenylenediamine compounds of the present invention may be present in various forms, for example as a powdered solid reagent, as a reagent immobilized on a bead or electrode surface, or as a solution in a suitable storage solution, such as a solution protected from light. can be provided.

An example of an electrochemical measurement is the measurement of glucose concentration. In the case of colorimetric electrochemical measurement, the glucose concentration can be measured, for example, as follows. Glucose dehydrogenase (GDH) was used in the reaction layer for electrochemical measurements, and N-(2-acetamido)imidodiacetic acid (ADA) and bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane (Bis) were used as reaction accelerators. -Tris), sodium carbonate, and imidazole. Here, a pH buffer and a coloring reagent (color-changing reagent) are added as necessary. A sample containing glucose is added to this and allowed to react for a certain period of time. During this time, the absorbance corresponding to the maximum absorption wavelength of the dye that is polymerized or reduced by receiving electrons directly from GDH is monitored. For the rate method, the rate of change in absorbance over time is used, and for the endpoint method, the change in absorbance up to the time when all the glucose in the sample has been oxidized is determined using a calibration prepared in advance using a glucose solution at a standard concentration. Based on the tion curve, the glucose concentration in the sample can be calculated.

As a coloring reagent (color-changing reagent) that can be used in this method, for example, 2,6-dichloroindophenol (DCIP) is added as an electron acceptor, and glucose can be quantified by monitoring the decrease in absorbance at 600 nm. . Furthermore, by adding nitrotetrazolium blue (NTB) as a coloring reagent and measuring the absorbance at 570 nm, it is possible to determine the amount of diformazan produced and calculate the glucose concentration. Needless to say, the coloring reagent (color-changing reagent) used is not limited to these.

In one embodiment, a sensor capable of detecting the compound of the present invention can be produced by utilizing the property of the compound of the present invention to be adsorbed to an electrode. An electrode, such as a carbon electrode, is inserted into a solution containing the compound of the present invention, such as DPPD, and then inserted into the solution or into another measurement solution after a certain period of time, and electrochemical measurements, such as CV or chronoamperometry, are carried out. The amount of the compound of the present invention can be detected quantitatively or qualitatively.

(enzyme sensor)
In one embodiment, the present invention provides an enzyme sensor comprising an electrode on which an oxidoreductase is immobilized and an electrode modifier of the present invention is adsorbed. Examples of the electrode of the enzyme sensor include a carbon electrode, a gold electrode, a platinum electrode, and the like, on which the redox enzyme can be coated or immobilized. Furthermore, conductive materials such as Co, Pd, Rh, Ir, Ru, Os, Re, Ni, Cr, Fe, Mo, Ti, Al, Cu, V, Nb, Zr, Sn, In, Ga, Mg, Pb, Fine metal particles containing at least one element among Au, Pt, and Ag may be included, and these may be alloyed or plated. Examples of carbon include carbon nanotubes, carbon black, graphite, fullerene, and derivatives thereof. For methods of immobilizing oxidoreductases, see the above paragraph of "Methods for immobilizing enzymes or oxidoreductases."

In certain embodiments, an example of an enzyme sensor of the present invention is a glucose sensor. This glucose sensor may have the phenylenediamine-based compound of the present invention adsorbed on the electrode and GDH or glucose oxidase (GOD) immobilized on the electrode. The glucose sensor can be used for continuous blood glucose measurements and continuous glucose monitoring.

The electrode, enzyme sensor, and electrochemical measurement composition of the present invention can be used in various electrochemical measurement techniques by using a potentiostat, a galvanostat, or the like. Electrochemical measurement methods include various techniques such as amperometry, such as chronoamperometry, potential step chronoamperometry, voltammetry, such as cyclic voltammetry, differential pulse voltammetry, potentiometry, and coulometry. For example, if the substrate to be measured is glucose, the glucose concentration in the sample can be calculated by measuring the current when glucose is reduced by amperometry. The applied voltage depends on the conditions and device settings, but can be, for example, -1000 mV to +1000 mV (vs. Ag/AgCl).

Note that whether or not the test compound is adsorbed to the electrode can be confirmed by cyclic voltammetry. The sweep speed is varied, for example in the range of 10 mV/sec to 50 mV/sec, and how the maximum value of oxidation current (I _Omax ) and the maximum value of reduction current (I _Rmax ) change is investigated. Generally, it is known that when a mediator is adsorbed to an electrode, the sweep speed of cyclic voltammetry and the values of I _Omax and I _Rmax are in a proportional relationship. When the mediator is diffused, I _Omax and I _Rmax are proportional to the 0.5th power of the sweep speed. From the relationship between I _Omax and I _Rmax and the sweep speed, it is determined whether the compound is adsorbed or diffused.

An example of electrochemical measurement is the electrochemical measurement of glucose. In certain embodiments, the present invention provides a method for generating glucose electricity, comprising contacting a sample that may contain glucose, a phenylenediamine-based compound, and purified glucose oxidase or purified glucose dehydrogenase, and measuring an electric current. Provides a chemical measurement method. The phenylenediamine compound may be present in solution or adsorbed on the electrode, and the enzyme may be immobilized on the electrode.

For example, electrochemical measurement of glucose concentration can be performed as follows. Fill the thermostatic cell with a buffer solution and maintain it at a constant temperature. An electrode on which GDH or GOD is immobilized is used as a working electrode, and a counter electrode (for example, a platinum electrode) and a reference electrode (for example, an Ag/AgCl electrode or an Ag/Ag+ electrode) are used. The phenylenediamine compound of the present invention is added to the reaction solution. After a constant voltage is applied to the carbon electrode and the current becomes steady, a sample containing glucose is added and the increase in current is measured. The glucose concentration in the sample can be calculated according to a calibration curve prepared using a standard concentration glucose solution. The potential to be applied can be, for example, +800mV or less, +700mV or less, +600mV or less, +500mV or less, +400mV or less, +300mV or less, +200mV or less, +100mV or less, +50mV or less, and -200mV or less. , -100mV or more, -50mV or more, for example 0mV or more, for example +800mV to -200mV, +800mV to -100mV, +800mV to -50mV, +600mV to 0mV, +500mV to 0mV, +400mV ~0mV, +300mV to 0mV, +200mV to 0mV (all relative to the silver-silver chloride reference electrode). The pH of the measurement solution containing glucose can be in the range of pH 3-10. For example, the pH is 5, pH 6, pH 7, pH 8, pH 9, or pH 10, and the solution may contain glycine, acetic acid, citric acid, phosphoric acid, carbonic acid, Good's buffer, etc. as a buffer. Even when enzymes other than GDH and GOD and substrates other than glucose are used, the pH can be changed as appropriate for measurement.

As a specific example, a phenylenediamine compound, such as IPPD, is immobilized on a glassy carbon (GC) electrode in advance, and then 0.2U to 150U, for example 0.5U to 100U of GDH or GOD is immobilized, and the response current to glucose concentration is Measure the value. Add 10.0 ml of 100 mM potassium phosphate buffer (pH 6.0) into the electrolysis cell. Connect the GC electrode to a potentiostat BAS100B/W (manufactured by BAS), stir the solution at 37 °C, and apply +500 mV to the silver-silver chloride reference electrode. Add 1M D-glucose solution to these systems to a final concentration of 5, 10, 20, 30, 40, and 50mM, and measure the steady-state current value after each addition. This current value is plotted against known glucose concentrations (5, 10, 20, 30, 40, 50 mM) to create a calibration curve. This makes it possible to quantify glucose using the GDH or GOD enzyme-immobilized electrode.

Furthermore, printed electrodes can also be used for electrochemical measurements. This makes it possible to reduce the amount of solution required for measurement. The electrodes can be formed on an insulating substrate. Specifically, the electrodes can be formed on the substrate by photolithography, screen printing, gravure printing, flexographic printing, or other printing techniques. Examples of the material for the insulating substrate include silicon, glass, ceramic, polyvinyl chloride, polyethylene, polypropylene, polyester, etc., and materials with high resistance to various solvents and chemicals can be used. The area of the working electrode can be set depending on the desired response current. For example, in some embodiments, the area of the working electrode is 1 mm ² or more, 1.5 mm ² or more, 2 mm ² or more, 2.5 mm ² or more, 3 mm ^{2 or more, 4 mm 2 or more, 5 mm 2 or more} , 6 mm ² or more, 7 mm ² or more, 8 mm ² or more, 9mm ² or more, 10mm ² or more, 12mm 2 or more, 15mm ² or more, 20mm ² or more, 30mm ² or more, 40mm ^{2 or} more, 50mm ² or more, 1cm 2 or more, 2cm ^{2 or} more, 3cm ² or more, 4cm ² or more , 5 cm ² or more, for example 10 cm ² or more. In some embodiments, the area of the working electrode can be less than or equal to 10 cm ² , such as less than or equal to 5 cm ² , such as less than or equal to 1 cm ² . The same can be said of the opposite. Furthermore, carbon nanotubes, graphene, Ketjenblack, etc. can be immobilized on the working electrode to increase the apparent surface area. In this case, the apparent area can increase by 10 times or more, 50 times or more, 100 times or more, or 1000 times or more.

In certain embodiments, the electrode modifier, electrode adsorbent, or electron transfer enhancer of the present invention, when used with an electrode, is 0.1 pmol or more, 0.2 pmol or more, 0.3 pmol or more, per 1 cm ² of working electrode area. 0.4 pmol or more, 0.5 pmol or more, 1 pmol or more, 10 mmol or less, 5 mmol or less, 1 mmol or less, 800 μmol or less, 600 μmol or less, 500 μmol or less, 400 μmol or less, 300 μmol or less, 200 μmol or less, 100 μmol or less, 50 μmol or less, for example 0.1 pmol to 10 mmol, 0.1 pmol to 5 mmol, 0.2 pmol to 1 mmol, 0.3 pmol to 800 μmol, 0.4 pmol to 600 μmol, 0.5 pmol to 500 μmol, 0.6 pmol to 400 μmol, 0.7 pmol to 300 μmol, 0.8 pmol to 200 μmol, 0.9 pmol to 100 μmol, 1 pmol to 50 μmol Can be used in quantity. These values are based on an assumption that the area of the working electrode is 1cm ² .If the area of the working electrode is increased or decreased, or if carbon nanotubes, graphene, etc. with a large specific surface area are used, the apparent surface area will increase. However, corresponding amounts of the electrode modifier of the invention may be used.

(Battery of the present invention)
In one embodiment, the present invention provides a battery comprising the phenylenediamine compound, electrode modifier, electrode adsorbent, or electron transfer enhancer of the present invention. In certain embodiments, in the battery of the present invention, the compound of the present invention is immobilized on the electrode of the battery. In certain embodiments, the cells of the invention include an oxidoreductase. In one embodiment, the oxidoreductase may be immobilized on an electrode of the battery. In one embodiment, the present invention provides a method of generating electricity using the battery of the present invention.

(Fuel cell of the present invention)
In one embodiment, the present invention provides an anode or cathode for a fuel cell comprising the phenylenediamine compound, electrode modifier, electrode adsorbent, or electron transfer promoter of the present invention, and a fuel cell equipped with the anode or cathode. do. In one embodiment, the present invention provides a power generation method using the phenylenediamine compound of the present invention, an electrode to which the phenylenediamine compound is adsorbed, and an oxidoreductase such as GDH or GOD is immobilized on an anode electrode. A power generation method using a corresponding substrate such as glucose as fuel is provided. If appropriate, when the oxidoreductase shown above is immobilized, a compound serving as a substrate for the immobilized oxidoreductase can be used as fuel.

In one embodiment, the fuel cell of the present invention includes an anode or a cathode to which the phenylenediamine compound of the present invention is adsorbed, a fuel tank, a cathode, an anode having an oxidoreductase, and an electrolyte. Furthermore, the fuel cell of the present invention can have a load resistor placed between the anode and the cathode, if necessary, and can be provided with wiring therefor. In some embodiments, the load resistor is part of the fuel cell of the present invention. In some embodiments, the load resistor is not part of the fuel cell of the present invention, and the fuel cell of the present invention is configured such that it can be connected to a suitable load resistor. In the fuel cell of the present invention, the oxidoreductase constitutes a part of the anode. For example, the oxidoreductase may be in close proximity to or in contact with the anode, may be immobilized, or may be adsorbed. The fuel tank contains a compound immobilized on the electrode that serves as a substrate for an oxidoreductase. For example, if glucose dehydrogenase is immobilized on the electrode, the fuel can be glucose. In certain embodiments, fuel cells of the present invention may have an ion exchange membrane separating the anode and cathode. Ion exchange membranes may have pores between 1 nm and 20 nm. The anode can be a common electrode such as a carbon electrode. For example, an electrode made of conductive carbonaceous material such as carbon black, graphite, or activated carbon, or an electrode made of metal such as gold or platinum can be used. Specific examples include carbon paper, carbon cloth, carbon felt, glassy carbon, and HOPG (highly oriented pyrolytic graphite). As the paired cathode, for example, an electrode catalyst commonly used in fuel cells such as platinum or platinum alloy, carbonaceous material such as carbon black, graphite, carbon cloth, carbon felt, activated carbon, or gold , an electrode supported on a conductor made of platinum, etc., or a conductor made of the electrode catalyst itself, such as platinum or a platinum alloy, is used as the cathode electrode, and an oxidizing agent (cathode side substrate, oxygen, etc.) is supplied to the electrode catalyst. It can take any form. In some embodiments, the electrodes of the present invention exclude mercury electrodes.

In another embodiment, a substrate-reducing enzyme electrode may be used as a cathode paired with an anode comprising a substrate-oxidizing enzyme electrode as described above. Examples of oxidoreductases that reduce oxidizing agents include known enzymes such as laccase and bilirubin oxidase. When using an oxidoreductase as a catalyst for reducing an oxidizing agent, a known electron transfer mediator may be used as necessary. The cathode mediator may be the same as or different from the anode mediator. Examples of the oxidizing agent include oxygen, hydrogen peroxide, and the like.

In some embodiments, an oxygen-selective membrane (e.g., a dimethylpolysiloxane membrane) is placed around the cathode electrode to avoid the influence of impurities (such as ascorbic acid or uric acid) that interfere with the electrode reaction at the cathode. Can be done.

The power generation method of the present invention includes the step of supplying a compound that serves as a substrate for an oxidoreductase and serves as a fuel to an anode having an oxidoreductase. When the anode containing the oxidoreductase is supplied with fuel, the substrate is oxidized and the oxidoreductase concomitantly transfers the generated electrons to an electron transfer mediator that mediates electron transfer between the oxidoreductase and the electrode, e.g. The electrons are transferred to the phenylenediamine compound, and the electrons are transferred to the conductive substrate (anode electrode) by the electron transfer mediator. Electric current is generated when electrons reach the cathode electrode from the anode electrode through wiring (external circuit).

Protons (H ⁺ ) generated in the above process move within the electrolyte solution to the cathode electrode. Then, at the cathode electrode, the protons that have moved from the anode in the electrolyte solution, the electrons that have moved from the anode side via an external circuit, and the oxidizing agent (cathode side substrate) such as oxygen or hydrogen peroxide react. water is produced. This can be used to generate electricity.

(Organic battery of the present invention)
In one embodiment, the present invention provides an organic battery having a phenylenediamine-based compound, an electrode modifier, an electrode adsorbent, or an electron transfer promoter of the present invention. The electrode modifier, electrode adsorbent, or electron transfer enhancer may be adsorbed to the electrode. Examples of electrode materials used in organic batteries include, but are not limited to, quinone, indigo derivatives, benzoquinone compounds having a methoxy group, indigo carmine, pentacentetron, and the like. Electrode materials used in organic radical batteries include electrode materials in which a compound having a nitroxy radical, such as 2,2,6,6-tetramethylpiperidine-N-oxyl, is bonded to a polymer, such as polymethacrylate or acrylate, and lithium. Examples include, but are not limited to. See, for example, Kobunshi, Vol. 54, December issue, 2005, p.886. In organic batteries, the electrode modifier of the present invention can be used as an electron mediator on the anode and cathode sides.

The electrode modifier containing the phenylenediamine compound of the present invention and the electrode to which the electrode modifier is adsorbed can be used for various electrochemical measurements. Further, the electrode can be used as an enzyme sensor by immobilizing an oxidoreductase. Further, the electrode modifying agent containing the phenylenediamine compound of the present invention and the electrode to which the electrode modifying agent is adsorbed can be used in fuel cells and organic cells. These are just examples, and the use of the electrode modifier containing the phenylenediamine compound of the present invention or the electrode to which the electrode modifier is adsorbed is not limited thereto.

The invention is further illustrated by the following examples. However, the technical scope of the present invention is not limited in any way by these examples.

Materials and Methods Materials and reagents, unless otherwise specified, are either commercially available or obtained or prepared according to techniques conventional in the art or procedures in known literature. Single-walled carbon nanotubes manufactured by Sigma, Meijo Nano Carbon, or Zeon Nano Technology were used. The multi-walled carbon nanotubes used were those manufactured by Sigma, Meijo Nano Carbon, or Kanto Kagaku. The carbon nanotubes used were appropriately dispersed by ultrasonication in an aqueous solution containing a low-molecular-weight surfactant, a water-soluble polymer, a water-soluble polysaccharide, and the like. As the surfactant, Triton X-100, sodium dodecyl sulfate, etc. are used, but the surfactant is not limited thereto. Compounds N-methyl-N'-phenyl-p-phenylenediamine, N-ethyl-N'-phenyl-p-phenylenediamine, N-isopropyl-N'-(4-aminophenyl)-p-phenylenediamine, 2- Amino-4-isopropylamino-diphenylamine and N-isopropyl-N'-(4-hydroxyphenyl)-p-phenylenediamine were obtained from NARD Research Institute, Inc. (NARD). The manufacturing procedure for these is as follows.

生成物はシリカゲルカラム（展開溶媒 ヘプタン/酢酸エチル）にて精製し、質量分析及びNMRにより確認した。化合物1-1の[M+H]⁺イオンのm/zは理論値199.1、実測値199.1であり、化合物1-2の[M+H]⁺イオンのm/zは理論値213.1、実測値213.1であった。 Using the scheme below, N-methyl-N'-phenyl-p-phenylenediamine (denoted as compound 1-1 in the scheme) and N-ethyl-N'-phenyl-p-phenylenediamine (in the scheme Compound 1-2) was synthesized.

The product was purified using a silica gel column (developing solvent: heptane/ethyl acetate) and confirmed by mass spectrometry and NMR. The m/z of the [M+H] ⁺ ion in compound 1-1 is the theoretical value 199.1 and the measured value 199.1, and the m/z of the [M+H] ⁺ ion in compound 1-2 is the theoretical value 213.1 and the measured value It was 213.1.

生成物はシリカゲルカラム（展開溶媒 ヘプタン/酢酸エチル）にて精製し、質量分析及びNMRにより確認した。化合物2-2の[M+H]⁺イオンのm/zは理論値242.1、実測値242.1であり、化合物2-3の[M+H]⁺イオンのm/zは理論値243.1、実測値243.0であった。 Using the scheme below, N-isopropyl-N'-(4-aminophenyl)-p-phenylenediamine (denoted as compound 2-2 in the scheme) and N-isopropyl-N'-(4-hydroxyphenyl) -p-phenylenediamine (denoted as compound 2-3 in the scheme) was synthesized.

The product was purified using a silica gel column (developing solvent: heptane/ethyl acetate) and confirmed by mass spectrometry and NMR. The m/z of the [M+H] ⁺ ion in compound 2-2 is the theoretical value 242.1 and the measured value 242.1, and the m/z of the [M+H] ⁺ ion in compound 2-3 is the theoretical value 243.1 and the measured value It was 243.0.

生成物はシリカゲルカラム（展開溶媒 ヘプタン/酢酸エチル）にて精製し、質量分析及びNMRにより確認した。化合物2-1の[M+H]⁺イオンのm/zは理論値242.1、実測値242.2であった。 2-Amino-4-isopropylamino-diphenylamine (denoted as compound 2-1 in the scheme) was synthesized using the scheme below.

The product was purified using a silica gel column (developing solvent: heptane/ethyl acetate) and confirmed by mass spectrometry and NMR. The m/z of the [M+H] ⁺ ion of compound 2-1 was a theoretical value of 242.1 and an actual value of 242.2.

Other compounds were commercially available from TCI or Sigma-Aldrich.

Example 1. Introduction of Mucor-derived GDH gene into host and confirmation of GDH activity The amino acid sequence of Mucor-derived GDH (MpGDH) described in Patent No. 4648993 is shown in SEQ ID NO: 1, and the base sequence is shown in SEQ ID NO: 2. A gene encoding a modified GDH (MpGDH-M2) in which each of the following mutations was introduced into MpGDH was obtained. The amino acid sequence of MpGDH-M2 is shown in SEQ ID NO: 3, and the base sequence of the gene is shown in SEQ ID NO: 4. A DNA construct was prepared by inserting the target gene, MpGDH-M2 gene, into the multiple cloning site of plasmid pUC19 using a conventional method. Specifically, pUC19 linearized Vector included in the In-Fusion HD Cloning Kit (Clontech) was used. At the In-Fusion Cloning Site located at the multi-cloning site of pUC19, the MpGDH-M1 gene was ligated using the above-mentioned In-Fusion HD Cloning Kit according to the protocol attached to the kit, and the construct plasmid (pUC19 -MpGDH-M2) was obtained.

These genes were expressed in Aspergillus sojae and their GDH activity was evaluated.

Specifically, for the purpose of obtaining MpGDH-M2, Double-joint PCR (Fungal Genetics and Biology, 2004, Vol. 41, p.973-981) was performed using the GDH gene, and the 5' A cassette consisting of arm region - pyrG gene - TEF1 promoter gene - flavin-binding GDH gene - 3' arm region was constructed, and the pyrG disrupted strain derived from Aspergillus sawjae NBRC4239 strain (48 bp upstream of pyrG gene, 896 bp coding region) was constructed using the following procedure. , downstream 240bp deletion strain). Note that the pyrG gene is a uracil auxotrophic marker. Conidia of the pyrG-disrupted strain derived from Aspergillus sawjae NBRC4239 were inoculated into 100 ml of a polypeptone dextrin liquid medium containing 20 mM uridine in a 500 ml Erlenmeyer flask, and after culturing with shaking at 30°C for about 20 hours, the bacterial cells were isolated. Recovered. Protoplasts were prepared from the collected cells. Transformation was performed by the protoplast PEG method using the obtained protoplasts and 20 μg of the target gene inserted DNA construct, and then Czapek-Dox minimal medium (Difco; pH 6) containing 0.5% (w/v) agar and 1.2 M sorbitol was used. ) and incubated at 30°C for 5 days or more to obtain transformed Aspergillus sojae as having colony forming ability.

The resulting transformed Aspergillus sojae was able to grow in uridine-free medium by introducing pyrG, a gene that complements uridine auxotrophy, and was selected as a strain in which the desired gene had been introduced. did it. From the obtained strains, the desired transformants were confirmed by PCR and selected.

Each GDH was produced using transformed Aspergillus sojae transformed with the MpGDH mutant gene.

DPY liquid medium (1% (w/v) polypeptone, 2% (w/v) dextrin, 0.5% (w/v) yeast extract, 0.5% (w/v) KH ₂ PO4, 0.05 in a 200 ml Erlenmeyer flask) Conidia of each strain were inoculated into 40 ml of %(w/v) _MgSO4.7H20 ; pH not adjusted, and cultured with shaking at 160 rpm at 30° _C for 4 days. Next, the bacterial cells were filtered from the culture after culturing, and the obtained medium supernatant fraction was concentrated and desalted to 10 mL using Amicon Ultra-15, 30K NMWL (manufactured by Millipore), and added to 20 mM phosphorus containing 150 mM NaCl. It was replaced with acid potassium buffer (pH 6.5). Subsequently, it was applied to HiLoad 26/60 Superdex 200pg (manufactured by GE Healthcare) equilibrated with 20mM potassium phosphate buffer (pH 6.5) containing 150mM NaCl, and eluted with the same buffer to show GDH activity. Fractions were collected to obtain a purified sample of MpGDH-M2. Note that this enzyme is bound to FAD through its FAD binding site (holoenzyme).

Example 2. Confirmation of adsorption between phenylenediamine compounds and carbon electrode Three types of phenylenediamine compounds (N-isopropyl-N'-phenyl-p-phenylenediamine (IPPD, manufactured by Tokyo Kasei Kogyo Co., Ltd., product code P0327), N,N '-Diphenyl-p-phenylenediamine (DPPD, manufactured by Tokyo Chemical Industry Co., Ltd., product code D0609), N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD, manufactured by Tokyo Chemical Industry Co., Ltd.) , product code D3331) was used to perform cyclic voltammetry (CV) using printed electrodes.Specifically, the SCREEN-PRINTED, which has a carbon working electrode (12.6 mm ² ) and a silver reference electrode printed on it, was performed using a printed electrode. 3 μl of IPPD 10% ethanol aqueous solution with a final concentration of 10 μg/ml was applied onto ELECTRODES (manufactured by DropSens, product number DRP-C110) and dried at room temperature.The electrode was then washed with ultrapure water and connected to the dedicated connector ( The printed electrode was used as the working electrode, the silver-silver chloride electrode (manufactured by BAS) was used as the reference electrode, and the platinum electrode (BAS) was used as the reference electrode. The counter electrode was immersed in 10 mL of 100 mM potassium phosphate buffer (pH 7.0).While stirring this solution at 650 rpm, the voltage was varied from -200 mV to +400 mV (vs.Ag/AgCl). Cyclic voltammetry was performed with _a sweep rate _of It is generally known that when the mediator is adsorbed to the electrode, the sweep speed of cyclic voltammetry and the values of I _Omax and I _Rmax are proportional to each other. If so, I _Omax and I _Rmax are proportional to the 0.5th power of the sweep speed.

Cyclic voltammetry was performed using IPPD, and the results of plotting the sweep speed, I _Omax , and I _Rmax are shown in Figure 1. As a result, it was confirmed that the sweep speed and I _Omax and I _Rmax were in a proportional relationship, indicating that IPPD was adsorbed to the carbon electrode.

The results of similar tests using DPPD and 6PPD instead of IPPD are shown in FIGS. 2 and 3. It was confirmed that the sweep speed and I _Omax and I _Rmax were in a proportional relationship for all compounds, indicating that they were adsorbed to the carbon electrode like IPPD.

Comparative example (p-phenylenediamine)
Note that when similar measurements were performed using p-phenylenediamine, which is already known to function as a mediator, instead of the diphenylamine compound of the present invention, neither oxidation waves nor reduction waves were observed. This is thought to be because p-phenylenediamine was peeled off from the electrode by washing the electrode with ultrapure water or immersing it in potassium phosphate buffer.

Place 10 μl of p-phenylenediamine 10% ethanol solution with a final concentration of 100 μg/ml and 10 μl of 100 mM potassium phosphate buffer (pH 7.0) on the above printed electrode, and apply the voltage in the range of -200 mV to +400 mV (vs.Ag/Ag+). When cyclic voltammetry was performed by sweeping the voltage, it was confirmed that I _Omax and I _Rmax were proportional to the 0.5th power of the sweep speed, as shown in FIGS. 4 and 5. This indicates that p-phenylenediamine is not adsorbed to the electrode but is diffused.

Example 3. Confirmation of adsorption between the compound of the present invention and carbon electrode Bindschedler's green leuco base (BGLB, manufactured by Tokyo Kasei Kogyo Co., Ltd., product code B0482, CAS number 637-31-0) or tris[4-(diethylamino)phenyl] Cyclic voltammetry (CV) with a printed electrode was performed using amine (TDPA, manufactured by Sigma-Aldrich, product code 556394, CAS number 5981-09-9). Specifically, BGLB with a final concentration of 1 mg/ml was applied to a SCREEN-PRINTED ELECTRODES (manufactured by DropSens, product number DRP-C110), which was printed with a carbon working electrode (12.6 mm ² ) and a silver reference electrode. Compounds were adsorbed using an ethanol solution. Thereafter, cyclic voltammetry was performed in the same manner as in Example 2.

FIG. 6 shows the results of plotting the sweep speed, I _Omax , and I _Rmax in measurements using BGLB. As a result, it was confirmed that the sweep speed and I _Omax and I _Rmax were in a proportional relationship, indicating that BGLB was adsorbed to the carbon electrode.

FIG. 7 shows the results of a similar test using TDPA instead of BGLB. Since TDPA has two redox waves, they are expressed as I _O1 , I _O2 , I _R1 and I _R2 in the figure. Even when TDPA was used, it was confirmed that the sweep speed and I _Omax and I _Rmax were in a proportional relationship, indicating that it adsorbed to the carbon electrode like BGLB.

The following compounds were prepared using the same method as above using a round carbon electrode (manufactured by Biodevice Technology, DEP-Chip EP-PP) or a SCREEN-PRINTED ELECTRODES (manufactured by DropSens, product number DRP-C110). It was also confirmed that the same adsorption occurred on the electrode.

Example 4. Electrochemical evaluation using phenylenediamine compounds
Cyclic voltammetry using printed electrodes was performed using FAD-dependent GDH and three phenylenediamine compounds (IPPD, DPPD, and 6PPD). Specifically, a carbon working electrode (2.64 mm ² ) and a round carbon electrode (manufactured by Biodevice Technology, DEP-Chip EP-PP) with a printed silver-silver chloride reference electrode were connected to a special connector. Connected to ALS Electrochemical Analyzer 814D (manufactured by BAS), 2 μl of 2000 U/ml FADGDH-AA (manufactured by Kikkoman Biochemifa, product number 60100) solution and 50 mM phosphoric acid containing 1.5 M potassium chloride were added. 8 μl of potassium buffer (pH 7.0) and 10 μl of IPPD 10% ethanol aqueous solution were placed on the electrode. The final concentration of IPPD was 2.5 μM. Then, cyclic voltammetry was performed by sweeping the voltage in the range of -200 mV to 400 mV (vs. Ag/AgCl). The sweep speed was 10 mV/sec. Subsequently, a glucose solution was added to the final concentration of 200 mM, and cyclic voltammetry was performed in the same manner.
Figure 8 shows the cyclic voltammogram during measurement. The observation of a response current to glucose indicated that IPPD functions as a mediator.
Next, connect the SCREEN-PRINTED ELECTRODES (manufactured by DropSens, product number DRP-110), which is printed with a carbon working electrode (12.6 mm ² ) and a silver-silver chloride reference electrode, to a special connector (manufactured by DropSens, DRP). -CAC), connect to ALS Electrochemical Analyzer 814D (manufactured by BAS), and connect 5 μl of 2000 U/ml Glucose Dehydrogenase (FAD-dependent) (manufactured by BBI, Product Code: GLD3, hereinafter referred to as GLD3) solution. , 20 μl of a 50 mM potassium phosphate buffer (pH 7.0) containing 1.5 M potassium chloride, and 25 μl of a 10% aqueous ethanol solution of a phenylenediamine compound were placed on the electrode. The final concentrations of the compounds were 2.5 μM for IPPD and 0.5 μM for DPPD and 6PPD. Then, cyclic voltammetry was performed by sweeping the voltage in the range of -200 mV to 400 mV (vs. Ag/AgCl). The sweep speed was 30 mV/sec. Subsequently, glucose solutions were added to achieve various final concentrations, and cyclic voltammetry was performed in the same manner. In addition, as a control experiment, cyclic voltammetry was performed in the same manner as above under conditions that did not contain a phenylenediamine compound.

Furthermore, plots of the relationship between the final glucose concentration and the oxidation current value at 300 mV when using IPPD, DPPD, and 6PPD are shown in FIGS. 9, 10, and 11. A response current to glucose is observed regardless of whether IPPD, DPPD, or 6PPD is used. On the other hand, no response current was observed when these compounds were not included, indicating that IPPD, DPPD, and 6PPD all function as mediators. Therefore, it was shown that it can also be used as an anode electrode.

Next, in the same way as above, FADGDH-AA (manufactured by Kikkoman Biochemifa, product number 60100) or Glucose Dehydrogenase (FAD-dependent) (manufactured by BBI, product code: GLD1, hereinafter referred to as GLD1) and IPPD were mixed. , cyclic voltammetry was performed by sweeping the voltage in the range of -200 mV to +400 mV (vs. Ag/AgCl). However, the final concentration of IPPD was 5 μM in combination with FADGDH-AA and 2.5 μM in combination with GLD1. The results are shown in FIGS. 12 and 13. A response current to glucose was observed only when IPPD was added, indicating that IPPD functions as a mediator. Therefore, it was shown that it can also be used as an anode electrode.

Subsequently, GOD from A. niger Type The results are shown in FIG. A response current to glucose was observed only when IPPD was added, indicating that IPPD functions as a mediator. Therefore, it was shown that it can also be used as an anode electrode.

Example 5. Stability test of electrodes with immobilized compound of the present invention and GDH
Cyclic voltammetry (CV) using a printed electrode was performed using FAD-dependent GDH and N,N'-diphenyl-1,4-phenylenediamine (DPPD, manufactured by Tokyo Kasei Kogyo Co., Ltd., product code D0609). Specifically, DPPD was first adsorbed onto SCREEN-PRINTED ELECTRODES (manufactured by DropSens, product number DRP-C110) using the same procedure as in Example 2, and the electrodes were washed with ultrapure water and then air-dried. Thereafter, 3 μl of 4 mg/ml FADGDH-AA (manufactured by Kikkoman Biochemifa, product number 60100) solution was applied to the electrode and air-dried again. Next, the electrode was exposed to the vapor of 25% glutaraldehyde solution (Fujifilm Wako Pure Chemical Industries, Ltd., Wako 1st grade, product number 079-00533) for 30 minutes, and then the electrode was washed with pure water to form the DPPD/GDH immobilized electrode. was created. This electrode was connected to the ALS Electrochemical Analyzer 814D (manufactured by BAS) using a dedicated connector (manufactured by DropSens, DRP-CAC), and was prepared using a 20mM potassium phosphate buffer (pH 7) containing glucose at a final concentration of 100mM. 0) Immersed in 10ml. Using the printed electrode as the working electrode, the silver-silver chloride electrode (manufactured by BAS) as the reference electrode, and the platinum electrode (manufactured by BAS) as the counter electrode, the voltage was varied from -200 mV to +400 mV (vs. Ag/AgCl) while stirring the solution at 750 rpm. ) CV was performed by sweeping the voltage over the range. The sweep speed was 30 mV/sec. Subsequently, the electrode was taken out of the solution, washed with ultrapure water, and then immersed in a new measurement solution with the same composition, and the same measurement was repeated a total of three times. For the oxidation current in the third CV at +150 mV, the difference between the measured values in the solution containing glucose and the blank solution without glucose was 149 nA. The response current was higher than that of the blank, indicating that DPPD functions as an electron transfer promoter. Furthermore, when comparing the first, second and third CVs, the current values hardly changed, indicating that DPPD and GDH were stably immobilized on the electrode without being desorbed into the solution. It was shown that

Comparative example (methylene blue)
When methylene blue (MB), which is already known as a mediator, was used instead of the compound of the present invention, no glucose-dependent response current was observed. On the other hand, in the test system in which FADGDH-AA and MB were added to the electrolyte solution, a glucose-dependent response current was confirmed. From the above, it was shown that MB was not adsorbed to the electrode and was peeled off during the electrode fabrication process.

Comparative example (1-methoxy-5-methylphenazinium methyl sulfate)
In addition, a similar test was conducted using 1-methoxy-5-methylphenazinium methyl sulfate (mPMS), which is already known as a mediator like MB. Regarding the oxidation current in the third CV at +150 mV, there was almost no difference in the measured value between the glucose-containing solution and the glucose-free blank solution, and the value was significantly lower than when DPPD was used. This indicates that mPMS was easily peeled off from the electrode and diffused into the solution.

Example 6. Regarding other oxidoreductases Next, we performed chronoamperometry using printed electrodes using Fructosyl-peptide Oxidase (FPOX-CE) (manufactured by Kikkoman Biochemifa, product number 60123, hereinafter referred to as FPOX-CE) and IPPD. I did it. Specifically, 5 μl of 0.79 U/ml FPOX-CE solution, 35 μl of 100 mM potassium phosphate buffer (pH 8.0) containing 3 M sodium chloride, and 5 μl of 10% ethanol solution of 10 ng/μl IPPD were applied to the electrode. I put it on top. Furthermore, 1 μl of 9 mM fructosylglycine solution was added. Then, +250 mV (vs. Ag/AgCl) was applied, and the response current value change was observed for 120 seconds.

When the values were recorded 10 seconds after the application, it was found that the response current value increased as the concentration of fructosylglycine increased, as shown in FIG. 15. Therefore, it was shown that IPPD also functions as a mediator for FPOX-CE. Therefore, it was shown that it can also be used as an anode electrode.

CV using printed electrodes was performed using FADGDH-AA, BGLB, and TDPA. Specifically, a compound/GDH immobilized electrode was prepared in the same manner as in Example 2, and CV was performed. However, the CV sweep range was from -200 mV to +600 mV (vs. Ag/AgCl), and measurements were also performed using a solution that did not contain glucose as a control experiment.

A comparison of oxidation current values at +300 mV when using BGLB and TDPA is shown in FIGS. 16 and 17. A glucose-dependent response current was observed regardless of whether BGLB or TDPA was used. This revealed that both BGLB and TDPA can be used as mediators when FADGDH-AA is used.

Subsequently, CV was performed using PQQ-dependent GDH, N-isopropyl-N'-phenyl-1,4-phenylenediamine (IPPD, manufactured by Tokyo Kasei Kogyo Co., Ltd., product code P0327), and BGLB. Specifically, the printed electrode DRP-C110 was connected to the ALS Electrochemical Analyzer 814D using a dedicated connector (DRP-CAC), and 0.8 mg/ml of Glucose dehydrogenase (PQQ-dependent) (manufactured by Toyobo Co., Ltd., product code GLD-321) solution, 20 μl of 50 mM potassium phosphate buffer (pH 6.8) containing 1.5 M potassium chloride, and 25 μl of a 10% ethanol solution of 5 μM compound were dropped onto the electrode. As the enzyme solution, 50 mM potassium phosphate buffer (pH 6.8) containing 1 mM calcium chloride and 0.1% polyoxyethylene (10) octylphenyl ether was used. Then, CV was performed by sweeping the voltage in the range of -200 mV to +400 mV (vs. Ag/AgCl). The sweep speed was 30 mV/sec. Subsequently, glucose solutions were added to achieve various final concentrations, and CV was performed in the same manner.

The relationship between the final glucose concentration and the oxidation current value at +300 mV is plotted in FIGS. 18 and 19. Glucose concentration-dependent current was observed when using either compound IPPD or BGLB, indicating that these compounds function as mediators of PQQ-dependent GDH.

Subsequently, chronoamperometry using NAD-dependent GDH and DPPD was performed. Specifically, the printed electrode DRP-C110 was connected to the ALS Electrochemical Analyzer 814D using a dedicated connector (DRP-CAC), and 5 mg/ml of Glucose dehydrogenase (NAD(P)-dependent) (manufactured by Toyobo Co., Ltd., Place 5 μl of product code GLD-311) solution, 15 μl of 150 mM potassium phosphate buffer (pH 8.0) containing 1.5 M potassium chloride, 5 μl of 50 mM-NAD aqueous solution, and 25 μl of 10% ethanol solution of 5 μM-DPPD on the electrode. dripped. A 150 mM potassium phosphate buffer (pH 8.0) was used as the enzyme lysis solution. A potential of +150 mV (vs. Ag/AgCl) was applied, and 60 seconds after the start of measurement, 5 μl of a 500 mM glucose solution was added to measure the current change. In addition, as a control experiment, a similar test was conducted using a measurement solution that did not contain DPPD.
FIG. 20 shows the difference in response current before and after adding glucose, at the time when the current value became constant after adding the glucose solution. In the presence of DPPD, the current value significantly increased with the addition of glucose, whereas in the absence of DPPD, almost no response to glucose was observed. These results revealed that DPPD also functions as a mediator in NAD-dependent GDH.

Subsequently, CV using lactate oxidase (LOD) and IPPD was performed. Specifically, the printed electrode DRP-C110 was connected to the ALS Electrochemical Analyzer 814D using a dedicated connector (DRP-CAC), and 5 μl of 5 mg/ml Lactate Oxidase (manufactured by Toyobo Co., Ltd., product code LCO-301) solution was added. , 20 μl of 1 M potassium phosphate buffer (pH 7.0) containing 1.5 M potassium chloride, and 25 μl of 10% ethanol solution of 5 μM-IPPD were dropped onto the electrode. A 1M potassium phosphate buffer (pH 7.0) was used as the enzyme lysis solution. Then, CV was performed by sweeping the voltage in the range of -200 mV to +400 mV (vs. Ag/AgCl). The sweep speed was 30 mV/sec. Subsequently, lactic acid solutions were added to achieve various final concentrations, and CV was performed in the same manner.

FIG. 21 shows a plot of the relationship between the final lactic acid concentration and the oxidation current value at +150 mV. An increase in response current dependent on lactate concentration was observed, indicating that IPPD functions as a mediator of LOD.

Subsequently, CV using fructose dehydrogenase (FDH) and IPPD was performed. It is known that FDH can directly transfer electrons to the electrode even in the absence of a mediator (i.e., has direct electron transfer ability), and that a substrate-dependent response current can be observed even in the absence of a mediator. It is an enzyme. Specifically, the printed electrode DRP-C110 was connected to the ALS Electrochemical Analyzer 814D using a dedicated connector (DRP-CAC), and 10 mg/ml of D-Fructose dehydrogenase (manufactured by Toyobo Co., Ltd., product code FCD-302) was added. 5 μl of the solution, 20 μl of 150 mM sodium acetate buffer (pH 4.5) containing 1.5 M potassium chloride, and 25 μl of 10% ethanol solution of 5 μM-IPPD were dropped onto the electrode. A 150 mM sodium acetate buffer (pH 4.5) containing 0.1% polyoxyethylene (10) octylphenyl ether was used as the enzyme lysis solution. Then, CV was performed by sweeping the voltage in the range of -200 mV to +400 mV (vs. Ag/AgCl). The sweep speed was 30 mV/sec. Subsequently, fructose solutions were added to achieve various final concentrations, and CV was performed in the same manner.

FIG. 22 shows a plot of the relationship between the final fructose concentration and the oxidation current value at +100 mV, assuming that the current when no fructose is added is 0. Since FDH has direct electron transfer ability, a fructose-dependent response current is observed even in the absence of IPPD. However, in the presence of IPPD, a larger current was observed than in the absence of IPPD. This indicates that IPPD functions as a mediator even in FDH, which has direct electron transfer ability.

Example 7. Glucose measurement using phenylenediamine compound/GDH immobilized sensor
3 μl of 5% polyethyleneimine (average molecular weight 10,000) was applied onto SCREEN-PRINTED ELECTRODES (manufactured by DropSens, product number DRP-C110) and dried at room temperature. Subsequently, 3 μl of IPPD (dissolved in 10% ethanol) and 30 U of GDH-M2 were applied and dried at room temperature. Finally, 1 μl of 2.5 mg/dl polyethylene glycol diglycidyl ether (average molecular weight 500) was applied and left at 4° C. overnight. The obtained electrode was washed with pure water and used as an IPPD/GDH immobilized electrode. Next, the IPPD/GDH immobilized electrode, Ag/AgCl reference electrode, and platinum counter electrode were immersed in 100 mM potassium phosphate buffer (pH 7), and CV measurements were performed. The sweeping speed was 10 mV/sec. 2M glucose was added as appropriate, and the oxidation current value at +200 mV at each glucose concentration was calculated. The results are shown in FIG. It was found that as the glucose concentration increased, the response current value also increased. Therefore, it can be said that the glucose concentration can be quantified by measuring glucose whose concentration is known and creating a calibration curve. It was also shown that it can be used as an anode electrode.

Example 8. Adsorption confirmation test when using electrode materials other than carbon
CV measurements were performed by immersing a gold electrode (manufactured by BAS, Cat No. 002421), an Ag/AgCl reference electrode, and a platinum counter electrode in a pH 7 phosphate buffer containing IPPD and GDH-M2. The sweep speed was set to 10 to 100 mV/sec, and the sweep speed, I _Omax , and I _Rmax were plotted. As a result, it was confirmed that the sweep speed and I _Omax and I _Rmax were in a proportional relationship, indicating that IPPD was adsorbed to the gold electrode.
Similarly, when a platinum electrode (manufactured by BAS, Cat No. 002422) was used instead of a gold electrode, it was confirmed that the sweep speed was proportional to I _Omax and I _Rmax , and the IPPD was It was shown to be adsorbed.

Example 9. Electrochemical Detection of Compounds of the Invention at Low Concentrations Printed electrodes DRP-C110 were placed in 50 mM potassium phosphate buffer containing DPPD diluted to various concentrations from 10 pM to 100 nM. As a control experiment, a solution containing no DPPD was also performed. After allowing it to stand as appropriate for 1 hour to 5 days, connect it to the ALS Electrochemical Analyzer 814D using the dedicated connector (DRP-CAC) and add 50 mM phosphoric acid containing FADGDH-AA at a final concentration of 4 mg/ml without DPPD. CV was performed in 10 ml of potassium buffer (pH 6.8) with a voltage sweep in the range of -400 mV to +200 mV (vs. Ag/Ag+). The sweep speed was 30 mV/sec. Thereafter, 1 ml of 1M glucose solution was added and CV was performed in the same manner, and the value was calculated by subtracting the oxidation current value at +100 mV in the absence of glucose addition from the oxidation current value at +100 mV.

FIG. 24 shows the results using a 10 pM DPPD solution. The current value increased upon addition of glucose, suggesting that this system is useful as a sensor capable of detecting DPPD even at concentrations as low as 10 pM. Note that when a solution containing no DPPD was used, no increase in response current was observed upon addition of glucose. Similarly, when DPPD of 100 pM, 1 nM, 10 nM, and 100 nM was used, a significant increase in response current was confirmed when glucose was added compared to when no glucose was added.

Example 10. Building a fuel cell
80 μl of the multi-walled carbon nanotube solution was applied in several portions to a 5 mm x 5 mm carbon cloth (manufactured by Toyo Technica) and dried at 60°C. After washing with pure water, it was further dried to adsorb and fix IPPD. Subsequently, 20 μl of 12 mg/ml FAD-dependent glucose dehydrogenase (GLD1, manufactured by Funakoshi) was applied and dried at 25°C. GLD1 was cross-linked and fixed by exposing the above carbon cloth to 25% glutaraldehyde vapor for 30 minutes, and it was used as an anode electrode. Platinum (manufactured by BAS) was used as a cathode electrode, and was immersed in PBS containing 100 mM D-glucose together with the above anode electrode and Ag/AgCl reference electrode, and connected to a variable resistor and potentiostat. When measured with open circuit potential, a current of 60μA/cm2 flowed when connected to 10kΩ. Note that when using an electrode that did not adsorb and fix IPPD, it was not possible to observe a current when connected to 10 kΩ. Therefore, by using the electrode on which the phenylenediamine compound of the present invention is immobilized, it is possible to produce a battery that can conduct electricity without adding a mediator to the fuel tank.

Furthermore, even when single-walled carbon nanotubes were used instead of multi-walled carbon nanotubes, current could be passed in the same way.

Example 11. Construction of a glucose sensor using the compound of the present invention A solution was prepared in which 1 mg/ml of IPPD was adsorbed and immobilized on single-walled carbon nanotubes. 3 μL of this solution was applied onto the working electrode of the printed electrode used in Example 2, dried, and thoroughly washed with ultrapure water. Subsequently, 4 mg/ml of GLD1 was applied and dried, and cross-linked and fixed with glutaraldehyde in the same manner as above. After washing with ultrapure water, it was immersed in PBS, a silver-silver chloride reference electrode and a platinum counter electrode were also immersed, and chronoamperometry measurement was performed by applying +250 mV. As a result, a response current of 1.3 μA was observed when 100 μM glucose was added compared to when no glucose was added. Therefore, it was suggested that an enzyme sensor could be similarly produced even if a printed electrode was coated with a solution in which carbon was adsorbed in advance. Note that a similar enzyme sensor could be fabricated using Ketjenblack instead of single-walled carbon nanotubes.

By using an electrode modifying agent containing the phenylenediamine compound of the present invention or an electrode to which the electrode modifying agent is adsorbed, various electrochemical measurements of batteries and the like can be performed.

All publications, patents, and patent applications mentioned or cited herein are incorporated by reference in their entirety.

Brief description of the sequence SEQ ID NO: 1 GDH from Mucor prainii (aa)
SEQ ID NO: 2 Mucor prainii-derived GDH (DNA)
SEQ ID NO: 3 MpGDH-M2 (aa)
SEQ ID NO: 4 MpGDH-M2 (DNA)

Claims (17)

A battery comprising an electrode modifier, the battery comprising:
The electrode modifier is an electrode modifier containing a compound that has the property of adsorbing to an electrode that has not been acid-treated,
The compound is a compound that has the property of being adsorbed to the electrode without being bonded to a polymer or being polymerized,
Adsorption of the compound to the electrode is confirmed by changing the sweep rate in cyclic voltammetry and by establishing a proportional relationship between the maximum value of oxidation current (I _Omax ) and the maximum value of reduction current (I _Rmax ). and
The compound has a structure of formula I or formula II,
[In the formula,
R ¹ is -NR ⁷ R ⁸ , -N=NR ⁹ , or -N ⁺ ≡N,
R ² is -NR ¹⁰ R ¹¹ or -N=NR ¹² ,
R ⁷ and R ⁸ are each independently hydrogen, a straight chain or branched C _1-7 alkyl, C _1-7 alkenyl, C _1- which may optionally be substituted with one or more X or V; ₇ alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, acetyl, carboxy, furanylformyl, pyrazolylformyl, 1-methyl-1H-pyrazol-5-ylformyl, 9,9 -dimethylfluoren-2-yl, -N ⁺ ≡N, -N=N-phenyl, benzyl,
and
R ¹⁰ is hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _3-9 optionally substituted by one or more X or V; cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl,
R ³ , R ⁴ , R ⁵ and R ⁶ are each independently hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl , optionally substituted with one or more Y; , C _1-7 alkynyl, C _1-7 alkoxy, halo, nitro, cyano, carboxy, sulfo, hydroxy or amino,
Alternatively, R ³ and R ⁴ or R ⁵ and R ⁶ together with the benzene ring containing them are optionally substituted with one or more oxo, X or W, or
, where * is bonded to the carbon atom to which R ³ is bonded, ** is bonded to the carbon atom to which R ⁴ is bonded, or * is bonded to the carbon atom to which R 5 is bonded, and ** is bonded to the carbon atom to which R ⁵ is bonded. is bonded to the carbon atom to which R ⁶ is bonded,
R ¹¹ is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X or Z;
R ⁹ is selected from the group consisting of hydrogen, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X;
R ¹² is phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, optionally substituted with one or more of X, W, isothiocyanate, or halosulfonyl;
selected from the group consisting of
Here, V is -O-acryloyl, acetylamino, or phenyl, which may be optionally substituted with C1-7 alkyl,
W is D or L-alanylsulfonyl, D or L-valylsulfonyl, D or L-leucylsulfonyl, D or L, optionally substituted with one or more amino, C1-7 alkyl, aminoalkyl -methionylsulfonyl, D or L-prolylsulfonyl, D or L-tryptophylsulfonyl, D or L-glycylsulfonyl, D or L-cysteinylsulfonyl, D or L-isoleuylsulfonyl, D or L-phenylalanylsulfonyl, D or L-tyrosylsulfonyl, D or L-serylsulfonyl, D or L-threonylsulfonyl, D or L-asparaginylsulfonyl, D or L-glutamylsulfonyl, D or L- Arginylsulfonyl, D or L-histidylsulfonyl, D or L-lysylsulfonyl, D or L-asparagylsulfonyl, D or L-glutaminylsulfonyl, -C(=O)-O-succinimidyl, acetyl, tri- fluoroacetyl, benzoylamino, -N=N-phenyl, phenylamino, diaminophenylazophenyl, or phenylazo optionally substituted by one or more X, naphthylazo optionally substituted by one or more Y, acetylamino,
and
X is a straight or branched chain, optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy, alkylamino, nitroso, nitro, and sulfo. is C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _1-7 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo, amino or alkylamino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, hydroxy, alkoxy and sulfo;
Z is -SO ₂ -CH=CH ₂ or -SO ₂ -C ₂ H ₄ -O-SO ₃ H, 4,6-dichlorotriazin-2-ylamino]
or a salt, anhydride or solvate thereof,
However, from the above compound, the structure of formula Ib
[In the formula,
R ^7b , R ^8b and R ^10b are each independently hydrogen, a linear or branched C _1-6 alkyl, C _1-6 alkenyl, C ₁ which may optionally be substituted with one or more Xb _-6 alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl, and R ^3b , R ^4b , R ^5b and R ^6b are each independently hydrogen, optionally 1 Straight chain or branched C _1-6 alkyl, C _1-6 alkenyl, C 1-6 alkynyl, _{C 1-6} alkoxy, _halo , nitro, cyano, carboxy, sulfo, which may be substituted by the above Y or amino;
R ^11b is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more Xb;
Here, Xb is a linear or branched C 1- which may be optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo _{. 6} alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _1-6 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo or amino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo]
Compounds having the structure, their salts, anhydrides, and solvates are excluded.
The battery. A battery comprising an electron transfer promoter,
The electron transfer promoter is an electron transfer promoter containing a compound that has the property of adsorbing to an electrode that has not been acid-treated,
The compound is a compound that has the property of being adsorbed to the electrode without being bonded to a polymer or being polymerized,
Adsorption of the compound to the electrode is confirmed by changing the sweep rate in cyclic voltammetry and by establishing a proportional relationship between the maximum value of oxidation current (I _Omax ) and the maximum value of reduction current (I _Rmax ). and
The compound has a structure of formula I or formula II,
[In the formula,
R ¹ is -NR ⁷ R ⁸ , -N=NR ⁹ , or -N ⁺ ≡N,
R ² is -NR ¹⁰ R ¹¹ or -N=NR ¹² ,
R ⁷ and R ⁸ are each independently hydrogen, a straight chain or branched C _1-7 alkyl, C _1-7 alkenyl, C _1- which may optionally be substituted with one or more X or V; ₇ alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, acetyl, carboxy, furanylformyl, pyrazolylformyl, 1-methyl-1H-pyrazol-5-ylformyl, 9,9 -dimethylfluoren-2-yl, -N ⁺ ≡N, -N=N-phenyl, benzyl,
and
R ¹⁰ is hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _3-9 optionally substituted by one or more X or V; cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl,
R ³ , R ⁴ , R ⁵ and R ⁶ are each independently hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl , optionally substituted with one or more Y; , C _1-7 alkynyl, C _1-7 alkoxy, halo, nitro, cyano, carboxy, sulfo, hydroxy or amino,
Alternatively, R ³ and R ⁴ or R ⁵ and R ⁶ together with the benzene ring containing them are optionally substituted with one or more oxo, X or W, or
, where * is bonded to the carbon atom to which R ³ is bonded, ** is bonded to the carbon atom to which R ⁴ is bonded, or * is bonded to the carbon atom to which R 5 is bonded, and ** is bonded to the carbon atom to which R ⁵ is bonded. is bonded to the carbon atom to which R ⁶ is bonded,
R ¹¹ is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X or Z;
R ⁹ is selected from the group consisting of hydrogen, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X;
R ¹² is phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, optionally substituted with one or more of X, W, isothiocyanate, or halosulfonyl;
selected from the group consisting of
Here, V is -O-acryloyl, acetylamino, or phenyl, which may be optionally substituted with C1-7 alkyl,
W is D or L-alanylsulfonyl, D or L-valylsulfonyl, D or L-leucylsulfonyl, D or L, optionally substituted with one or more amino, C1-7 alkyl, aminoalkyl -methionylsulfonyl, D or L-prolylsulfonyl, D or L-tryptophylsulfonyl, D or L-glycylsulfonyl, D or L-cysteinylsulfonyl, D or L-isoleuylsulfonyl, D or L-phenylalanylsulfonyl, D or L-tyrosylsulfonyl, D or L-serylsulfonyl, D or L-threonylsulfonyl, D or L-asparaginylsulfonyl, D or L-glutamylsulfonyl, D or L- Arginylsulfonyl, D or L-histidylsulfonyl, D or L-lysylsulfonyl, D or L-asparagylsulfonyl, D or L-glutaminylsulfonyl, -C(=O)-O-succinimidyl, acetyl, tri- fluoroacetyl, benzoylamino, -N=N-phenyl, phenylamino, diaminophenylazophenyl, or phenylazo optionally substituted by one or more X, naphthylazo optionally substituted by one or more Y, acetylamino,
and
X is a straight or branched chain, optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy, alkylamino, nitroso, nitro, and sulfo. is C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _1-7 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo, amino or alkylamino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, hydroxy, alkoxy and sulfo;
Z is -SO ₂ -CH=CH ₂ or -SO ₂ -C ₂ H ₄ -O-SO ₃ H, 4,6-dichlorotriazin-2-ylamino]
or a salt, anhydride or solvate thereof,
However, from the above compound, the structure of formula Ib
[In the formula,
R ^7b , R ^8b and R ^10b are each independently hydrogen, a linear or branched C _1-6 alkyl, C _1-6 alkenyl, C ₁ which may optionally be substituted with one or more Xb _-6 alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl, and R ^3b , R ^4b , R ^5b and R ^6b are each independently hydrogen, optionally 1 Straight chain or branched C _1-6 alkyl, C _1-6 alkenyl, C 1-6 alkynyl, _{C 1-6} alkoxy, _halo , nitro, cyano, carboxy, sulfo, which may be substituted by the above Y or amino;
R ^11b is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more Xb;
Here, Xb is a linear or branched C 1- which may be optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo _{. 6} alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _1-6 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo or amino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo]
Compounds having the structure, their salts, anhydrides, and solvates are excluded.
The battery. The battery according to claim 1, wherein the compound is immobilized on an electrode of the battery. The battery according to any one of claims 1 to 3, comprising an oxidoreductase. The battery according to claim 4, wherein the oxidoreductase is immobilized on the electrode. A composition comprising an electron transfer enhancer,
The electron transfer promoter is an electrode modifying agent containing a compound that has the property of adsorbing to a non-acid-treated electrode,
The compound is a compound that has the property of being adsorbed to the electrode without being bonded to a polymer or being polymerized,
Adsorption of the compound to the electrode is confirmed by changing the sweep rate in cyclic voltammetry and by establishing a proportional relationship between the maximum value of oxidation current (I _Omax ) and the maximum value of reduction current (I _Rmax ). and
The compound has a structure of formula I or formula II,
[In the formula,
R ¹ is -NR ⁷ R ⁸ , -N=NR ⁹ , or -N ⁺ ≡N,
R ² is -NR ¹⁰ R ¹¹ or -N=NR ¹² ,
R ⁷ and R ⁸ are each independently hydrogen, a straight chain or branched C _1-7 alkyl, C _1-7 alkenyl, C _1- which may optionally be substituted with one or more X or V; ₇ alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, acetyl, carboxy, furanylformyl, pyrazolylformyl, 1-methyl-1H-pyrazol-5-ylformyl, 9,9 -dimethylfluoren-2-yl, -N ⁺ ≡N, -N=N-phenyl, benzyl,
and
R ¹⁰ is hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _3-9 optionally substituted by one or more X or V; cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl,
R ³ , R ⁴ , R ⁵ and R ⁶ are each independently hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl , optionally substituted with one or more Y; , C _1-7 alkynyl, C _1-7 alkoxy, halo, nitro, cyano, carboxy, sulfo, hydroxy or amino,
Alternatively, R ³ and R ⁴ or R ⁵ and R ⁶ together with the benzene ring containing them are optionally substituted with one or more oxo, X or W, or
, where * is bonded to the carbon atom to which R ³ is bonded, ** is bonded to the carbon atom to which R ⁴ is bonded, or * is bonded to the carbon atom to which R 5 is bonded, and ** is bonded to the carbon atom to which R ⁵ is bonded. is bonded to the carbon atom to which R ⁶ is bonded,
R ¹¹ is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X or Z;
R ⁹ is selected from the group consisting of hydrogen, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X;
R ¹² is phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, optionally substituted with one or more of X, W, isothiocyanate, or halosulfonyl;
selected from the group consisting of
Here, V is -O-acryloyl, acetylamino, or phenyl, which may be optionally substituted with C1-7 alkyl,
W is D or L-alanylsulfonyl, D or L-valylsulfonyl, D or L-leucylsulfonyl, D or L, optionally substituted with one or more amino, C1-7 alkyl, aminoalkyl -methionylsulfonyl, D or L-prolylsulfonyl, D or L-tryptophylsulfonyl, D or L-glycylsulfonyl, D or L-cysteinylsulfonyl, D or L-isoleuylsulfonyl, D or L-phenylalanylsulfonyl, D or L-tyrosylsulfonyl, D or L-serylsulfonyl, D or L-threonylsulfonyl, D or L-asparaginylsulfonyl, D or L-glutamylsulfonyl, D or L- Arginylsulfonyl, D or L-histidylsulfonyl, D or L-lysylsulfonyl, D or L-asparagylsulfonyl, D or L-glutaminylsulfonyl, -C(=O)-O-succinimidyl, acetyl, tri- fluoroacetyl, benzoylamino, -N=N-phenyl, phenylamino, diaminophenylazophenyl, or phenylazo optionally substituted by one or more X, naphthylazo optionally substituted by one or more Y, acetylamino,
and
X is a straight or branched chain, optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy, alkylamino, nitroso, nitro, and sulfo. is C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _1-7 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo, amino or alkylamino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, hydroxy, alkoxy and sulfo;
Z is -SO ₂ -CH=CH ₂ or -SO ₂ -C ₂ H ₄ -O-SO ₃ H, 4,6-dichlorotriazin-2-ylamino]
or a salt, anhydride or solvate thereof,
However, from the above compound, the structure of formula Ib
[In the formula,
R ^7b , R ^8b and R ^10b are each independently hydrogen, a linear or branched C _1-6 alkyl, C _1-6 alkenyl, C ₁ which may optionally be substituted with one or more Xb _-6 alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl, and R ^3b , R ^4b , R ^5b and R ^6b are each independently hydrogen, optionally 1 Straight chain or branched C _1-6 alkyl, C _1-6 alkenyl, C 1-6 alkynyl, _{C 1-6} alkoxy, _halo , nitro, cyano, carboxy, sulfo, which may be substituted by the above Y or amino;
R ^11b is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more Xb;
Here, Xb is a linear or branched C 1- which may be optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo _{. 6} alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _1-6 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo or amino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo]
Compounds having the structure, their salts, anhydrides, and solvates are excluded,
Furthermore, from the compound
and its salts, anhydrides, and solvates are excluded, and from said compounds, N-methyl-N-phenyl-1,4-phenylenediamine and its salts, anhydrides, and solvates are excluded. removed,
Said composition. An electrode comprising an electron transfer enhancer,
The electron transfer promoter is an electrode modifying agent containing a compound that has the property of adsorbing to a non-acid-treated electrode,
The compound is a compound that has the property of being adsorbed to the electrode without being bonded to a polymer or being polymerized,
Adsorption of the compound to the electrode is confirmed by changing the sweep rate in cyclic voltammetry and by establishing a proportional relationship between the maximum value of oxidation current (I _Omax ) and the maximum value of reduction current (I _Rmax ). and
The compound has a structure of formula I or formula II,
[In the formula,
R ¹ is -NR ⁷ R ⁸ , -N=NR ⁹ , or -N ⁺ ≡N,
R ² is -NR ¹⁰ R ¹¹ or -N=NR ¹² ,
R ⁷ and R ⁸ are each independently hydrogen, a straight chain or branched C _1-7 alkyl, C _1-7 alkenyl, C _1- which may optionally be substituted with one or more X or V; ₇ alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, acetyl, carboxy, furanylformyl, pyrazolylformyl, 1-methyl-1H-pyrazol-5-ylformyl, 9,9 -dimethylfluoren-2-yl, -N ⁺ ≡N, -N=N-phenyl, benzyl,
and
R ¹⁰ is hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _3-9 optionally substituted by one or more X or V; cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl,
R ³ , R ⁴ , R ⁵ and R ⁶ are each independently hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl , optionally substituted with one or more Y; , C _1-7 alkynyl, C _1-7 alkoxy, halo, nitro, cyano, carboxy, sulfo, hydroxy or amino,
Alternatively, R ³ and R ⁴ or R ⁵ and R ⁶ together with the benzene ring containing them are optionally substituted with one or more oxo, X or W, or
, where * is bonded to the carbon atom to which R ³ is bonded, ** is bonded to the carbon atom to which R ⁴ is bonded, or * is bonded to the carbon atom to which R 5 is bonded, and ** is bonded to the carbon atom to which R ⁵ is bonded. is bonded to the carbon atom to which R ⁶ is bonded,
R ¹¹ is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X or Z;
R ⁹ is selected from the group consisting of hydrogen, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X;
R ¹² is phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, optionally substituted with one or more of X, W, isothiocyanate, or halosulfonyl;
selected from the group consisting of
Here, V is -O-acryloyl, acetylamino, or phenyl, which may be optionally substituted with C1-7 alkyl,
W is D or L-alanylsulfonyl, D or L-valylsulfonyl, D or L-leucylsulfonyl, D or L, optionally substituted with one or more amino, C1-7 alkyl, aminoalkyl -methionylsulfonyl, D or L-prolylsulfonyl, D or L-tryptophylsulfonyl, D or L-glycylsulfonyl, D or L-cysteinylsulfonyl, D or L-isoleuylsulfonyl, D or L-phenylalanylsulfonyl, D or L-tyrosylsulfonyl, D or L-serylsulfonyl, D or L-threonylsulfonyl, D or L-asparaginylsulfonyl, D or L-glutamylsulfonyl, D or L- Arginylsulfonyl, D or L-histidylsulfonyl, D or L-lysylsulfonyl, D or L-asparagylsulfonyl, D or L-glutaminylsulfonyl, -C(=O)-O-succinimidyl, acetyl, tri- fluoroacetyl, benzoylamino, -N=N-phenyl, phenylamino, diaminophenylazophenyl, or phenylazo optionally substituted by one or more X, naphthylazo optionally substituted by one or more Y, acetylamino,
and
X is a straight or branched chain, optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy, alkylamino, nitroso, nitro, and sulfo. is C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _1-7 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo, amino or alkylamino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, hydroxy, alkoxy and sulfo;
Z is -SO ₂ -CH=CH ₂ or -SO ₂ -C ₂ H ₄ -O-SO ₃ H, 4,6-dichlorotriazin-2-ylamino]
or a salt, anhydride or solvate thereof,
However, from the above compound, the structure of formula Ib
[In the formula,
R ^7b , R ^8b and R ^10b are each independently hydrogen, a linear or branched C _1-6 alkyl, C _1-6 alkenyl, C ₁ which may optionally be substituted with one or more Xb _-6 alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl, and R ^3b , R ^4b , R ^5b and R ^6b are each independently hydrogen, optionally 1 Straight chain or branched C _1-6 alkyl, C _1-6 alkenyl, C 1-6 alkynyl, _{C 1-6} alkoxy, _halo , nitro, cyano, carboxy, sulfo, which may be substituted by the above Y or amino;
R ^11b is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more Xb;
Here, Xb is a linear or branched C 1- which may be optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo _{. 6} alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _1-6 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo or amino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo]
Compounds having the structure, their salts, anhydrides, and solvates are excluded,
Furthermore, from the compound
and its salts, anhydrides, and solvates are excluded, and from said compounds, N-methyl-N-phenyl-1,4-phenylenediamine and its salts, anhydrides, and solvates are excluded. removed,
The electrode. The electrode according to claim 7, comprising an enzyme, or the composition according to claim 6, comprising an enzyme. The electrode or composition according to claim 8, wherein the enzyme is an oxidoreductase. The electrode according to claim 9, wherein an oxidoreductase is immobilized. A sensor comprising an electrode modifier, the sensor comprising:
The electrode modifier is an electrode modifier containing a compound that has the property of adsorbing to an electrode that has not been acid-treated,
The compound is a compound that has the property of being adsorbed to the electrode without being bonded to a polymer or being polymerized,
Adsorption of the compound to the electrode is confirmed by changing the sweep rate in cyclic voltammetry and by establishing a proportional relationship between the maximum value of oxidation current (I _Omax ) and the maximum value of reduction current (I _Rmax ). and
The compound has a structure of formula I or formula II,
[In the formula,
R ¹ is -NR ⁷ R ⁸ , -N=NR ⁹ , or -N ⁺ ≡N,
R ² is -NR ¹⁰ R ¹¹ or -N=NR ¹² ,
R ⁷ and R ⁸ are each independently hydrogen, a straight chain or branched C _1-7 alkyl, C _1-7 alkenyl, C _1- which may optionally be substituted with one or more X or V; ₇ alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, acetyl, carboxy, furanylformyl, pyrazolylformyl, 1-methyl-1H-pyrazol-5-ylformyl, 9,9 -dimethylfluoren-2-yl, -N ⁺ ≡N, -N=N-phenyl, benzyl,
and
R ¹⁰ is hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _3-9 optionally substituted by one or more X or V; cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl,
R ³ , R ⁴ , R ⁵ and R ⁶ are each independently hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl , optionally substituted with one or more Y; , C _1-7 alkynyl, C _1-7 alkoxy, halo, nitro, cyano, carboxy, sulfo, hydroxy or amino,
Alternatively, R ³ and R ⁴ or R ⁵ and R ⁶ together with the benzene ring containing them are optionally substituted with one or more oxo, X or W, or
, where * is bonded to the carbon atom to which R ³ is bonded, ** is bonded to the carbon atom to which R ⁴ is bonded, or * is bonded to the carbon atom to which R 5 is bonded, and ** is bonded to the carbon atom to which R ⁵ is bonded. is bonded to the carbon atom to which R ⁶ is bonded,
R ¹¹ is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X or Z;
R ⁹ is selected from the group consisting of hydrogen, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X;
R ¹² is phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, optionally substituted with one or more of X, W, isothiocyanate, or halosulfonyl;
selected from the group consisting of
Here, V is -O-acryloyl, acetylamino, or phenyl, which may be optionally substituted with C1-7 alkyl,
W is D or L-alanylsulfonyl, D or L-valylsulfonyl, D or L-leucylsulfonyl, D or L, optionally substituted with one or more amino, C1-7 alkyl, aminoalkyl -methionylsulfonyl, D or L-prolylsulfonyl, D or L-tryptophylsulfonyl, D or L-glycylsulfonyl, D or L-cysteinylsulfonyl, D or L-isoleuylsulfonyl, D or L-phenylalanylsulfonyl, D or L-tyrosylsulfonyl, D or L-serylsulfonyl, D or L-threonylsulfonyl, D or L-asparaginylsulfonyl, D or L-glutamylsulfonyl, D or L- Arginylsulfonyl, D or L-histidylsulfonyl, D or L-lysylsulfonyl, D or L-asparagylsulfonyl, D or L-glutaminylsulfonyl, -C(=O)-O-succinimidyl, acetyl, tri- fluoroacetyl, benzoylamino, -N=N-phenyl, phenylamino, diaminophenylazophenyl, or phenylazo optionally substituted by one or more X, naphthylazo optionally substituted by one or more Y, acetylamino,
and
X is a straight or branched chain, optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy, alkylamino, nitroso, nitro, and sulfo. is C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _1-7 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo, amino or alkylamino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, hydroxy, alkoxy and sulfo;
Z is -SO ₂ -CH=CH ₂ or -SO ₂ -C ₂ H ₄ -O-SO ₃ H, 4,6-dichlorotriazin-2-ylamino]
or a salt, anhydride or solvate thereof,
However, from the above compound, the structure of formula Ib
[In the formula,
R ^7b , R ^8b and R ^10b are each independently hydrogen, a linear or branched C _1-6 alkyl, C _1-6 alkenyl, C ₁ which may optionally be substituted with one or more Xb _-6 alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl, and R ^3b , R ^4b , R ^5b and R ^6b are each independently hydrogen, optionally 1 Straight chain or branched C _1-6 alkyl, C _1-6 alkenyl, C 1-6 alkynyl, _{C 1-6} alkoxy, _halo , nitro, cyano, carboxy, sulfo, which may be substituted by the above Y or amino;
R ^11b is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more Xb;
Here, Xb is a linear or branched C 1- which may be optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo _{. 6} alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _1-6 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo or amino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo]
Compounds having the structure, their salts, anhydrides, and solvates are excluded,
Furthermore, from the compound
and its salts, anhydrides, and solvates are excluded, and from said compounds, N-methyl-N-phenyl-1,4-phenylenediamine and its salts, anhydrides, and solvates are excluded. said sensor being excluded;
Or
An enzyme sensor comprising the electrode according to claim 9 or 10. The compound has a structure of formula Ia or IIa
[In the formula,
R ^1a is -NR ^7a R ^8a , -N=NR ^9a , or -N ⁺ ≡N,
R ^2a is -NR ^10a R ^11a or -N=NR ^12a ,
R ^7a and R ^8a are each independently hydrogen, linear or branched C _1-6 alkyl, C _1-6 alkenyl, C _1-6 alkynyl, optionally substituted with one or more Xa; , C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, or phenanthrenyl, and R ¹⁰ is hydrogen, optionally substituted by one or more Xa, straight or branched chain is C _1-6 alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl,
R ^3a , R ^4a , R ^5a and R ^6a are each independently hydrogen, linear or branched C _1-6 alkyl, C _1-6 alkenyl, optionally substituted with one or more Y; , C _1-6 alkynyl, C _1-6 alkoxy, halo, nitro, cyano, carboxy, sulfo, hydroxy or amino,
Alternatively, R ^3a and R ^4a or R ^5a and R ^6a are a benzene ring, or
, where * is bonded to the carbon atom to which R ^3a is bonded, ** is bonded to the carbon atom to which R ^4a is bonded, or * is bonded to the carbon atom to which R ^{5a is bonded, and ** is bonded to the carbon atom to which R 5a} is bonded. is bonded to the carbon atom to which R ^6a is bonded,
R ^11a is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more Xa;
R ^9a and R ^12a are each independently selected from the group consisting of hydrogen, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted with one or more Xa;
Xa is a linear or branched chain, optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, hydroxy, alkoxy, alkylamino, nitroso, nitro and sulfo. branched C _1-6 alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _1-6 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo, amino or alkylamino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo]
or has the structure IIb
[In the formula,
R ^7b , R ^8b and R ^10b are each independently hydrogen, a linear or branched C _1-6 alkyl, C _1-6 alkenyl, C ₁ which may optionally be substituted with one or more Xb _-6 alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl, and R ^3b , R ^4b , R ^5b and R ^6b are each independently hydrogen, optionally 1 Straight chain or branched C _1-6 alkyl, C _1-6 alkenyl, C _{1-6 alkynyl, C 1-6 alkoxy} , halo, nitro, cyano, carboxy, sulfo, which may be substituted by the above Y or amino;
R ^11b is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more Xb;
Here, Xb is a linear or branched C 1- which may be optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo _{. 6} alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _1-6 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo or amino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo]
or a salt, anhydride or solvate thereof, the battery according to any one of claims 1 to 5 , the composition according to claims 6, 8 or 9 , claims 7 to 10 or the sensor according to claim 11 . The compound is
13. The battery, composition, electrode or sensor according to claim 12 , selected from the group consisting of: An electrode modifier comprising a compound that has the property of adsorbing to an electrode that has not been acid-treated,
The compound is a compound that has the property of being adsorbed to the electrode without being bonded to a polymer or being polymerized,
Adsorption of the compound to the electrode is confirmed by changing the sweep rate in cyclic voltammetry and observing that the maximum value of oxidation current (I _Omax ) and the maximum value of reduction current (I _Rmax ) become proportional to each other. ,
A method for manufacturing a battery, the method comprising the step of bringing the electrode modifier into contact with an electrode of the battery ,
The compound has a structure of formula I or formula II,
[In the formula,
R ¹ is -NR ⁷ R ⁸ , -N=NR ⁹ , or -N ⁺ ≡N,
R ² is -NR ¹⁰ R ¹¹ or -N=NR ¹² ,
R ⁷ and R ⁸ are each independently hydrogen, a straight chain or branched C _1-7 alkyl, C _1-7 alkenyl, C _1- which may optionally be substituted with one or more X or V; ₇ alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, acetyl, carboxy, furanylformyl, pyrazolylformyl, 1-methyl-1H-pyrazol-5-ylformyl, 9,9 -dimethylfluoren-2-yl, -N ⁺ ≡N, -N=N-phenyl, benzyl,
and
R ¹⁰ is hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _3-9 optionally substituted by one or more X or V; cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl,
R ³ , R ⁴ , R ⁵ and R ⁶ are each independently hydrogen, linear or branched C _1-7 alkyl, C _1-7 alkenyl , optionally substituted with one or more Y; , C _1-7 alkynyl, C _1-7 alkoxy, halo, nitro, cyano, carboxy, sulfo, hydroxy or amino,
Alternatively, R ³ and R ⁴ or R ⁵ and R ⁶ together with the benzene ring containing them are optionally substituted with one or more oxo, X or W, or
, where * is bonded to the carbon atom to which R ³ is bonded, ** is bonded to the carbon atom to which R ⁴ is bonded, or * is bonded to the carbon atom to which R 5 is bonded, and ** is bonded to the carbon atom to which R ⁵ is bonded. is bonded to the carbon atom to which R ⁶ is bonded,
R ¹¹ is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X or Z;
R ⁹ is selected from the group consisting of hydrogen, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more X;
R ¹² is phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, optionally substituted with one or more of X, W, isothiocyanate, or halosulfonyl;
selected from the group consisting of
Here, V is -O-acryloyl, acetylamino, or phenyl, which may be optionally substituted with C1-7 alkyl,
W is D or L-alanylsulfonyl, D or L-valylsulfonyl, D or L-leucylsulfonyl, D or L, optionally substituted with one or more amino, C1-7 alkyl, aminoalkyl -methionylsulfonyl, D or L-prolylsulfonyl, D or L-tryptophylsulfonyl, D or L-glycylsulfonyl, D or L-cysteinylsulfonyl, D or L-isoleuylsulfonyl, D or L-phenylalanylsulfonyl, D or L-tyrosylsulfonyl, D or L-serylsulfonyl, D or L-threonylsulfonyl, D or L-asparaginylsulfonyl, D or L-glutamylsulfonyl, D or L- Arginylsulfonyl, D or L-histidylsulfonyl, D or L-lysylsulfonyl, D or L-asparagylsulfonyl, D or L-glutaminylsulfonyl, -C(=O)-O-succinimidyl, acetyl, tri- fluoroacetyl, benzoylamino, -N=N-phenyl, phenylamino, diaminophenylazophenyl, or phenylazo optionally substituted by one or more X, naphthylazo optionally substituted by one or more Y, acetylamino,
and
X is a straight or branched chain, optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy, alkylamino, nitroso, nitro, and sulfo. is C _1-7 alkyl, C _1-7 alkenyl, C _1-7 alkynyl, C _1-7 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo, amino or alkylamino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, hydroxy, alkoxy and sulfo;
Z is -SO ₂ -CH=CH ₂ or -SO ₂ -C ₂ H ₄ -O-SO ₃ H, 4,6-dichlorotriazin-2-ylamino]
or a salt, anhydride or solvate thereof,
However, from the above compound, the structure of formula Ib
[In the formula,
R ^7b , R ^8b and R ^10b are each independently hydrogen, a linear or branched C _1-6 alkyl, C _1-6 alkenyl, C ₁ which may optionally be substituted with one or more Xb _-6 alkynyl, C _3-9 cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, anthracenyl or phenanthrenyl, and R ^3b , R ^4b , R ^5b and R ^6b are each independently hydrogen, optionally 1 Straight chain or branched C _1-6 alkyl, C _1-6 alkenyl, C 1-6 alkynyl, _{C 1-6} alkoxy, _halo , nitro, cyano, carboxy, sulfo, which may be substituted by the above Y. or amino;
R ^11b is selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, anthracenyl and phenanthrenyl, optionally substituted by one or more Xb;
Here, Xb is a linear or branched C 1- which may be optionally substituted with one or more substituents selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo _{. 6} alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _1-6 alkoxy, halo, hydroxy, nitro, carboxy, cyano, sulfo or amino,
Y is selected from the group consisting of halo, amino, cyano, carboxy, carbonyl, alkoxy and sulfo]
Compounds having the structure, their salts, anhydrides, and solvates are excluded.
The manufacturing method . A power generation method using the battery according to any one of claims 1 to 5 and 12 to 13 . The composition according to any one of claims 6, 8, 9 and 12 to 13 ,
An electrochemical measurement method using the electrode according to any one of claims 7 to 10 and 12 to 13 , or the sensor according to any one of claims 11 to 13 . A method of altering or modifying an electrode, comprising the step of contacting the electrode with a composition according to any one of claims 6, 8, 9 and 12-13 .

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