JP2006219453A - Metal-discrimination-type two-color fluorescent molecule containing quinoline ring as parent nucleus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new fluorescent molecule discriminating a specific metal by fluorometric analysis of metal ion and generating new fluorescent light by the addition of the metal and provide a method for the use of the fluorescent molecule. <P>SOLUTION: The fluorescent molecule is a new quinoline compound or naphthalene compound expressed by general formula (I) (R<SP>1</SP>is a heterocyclic group or a carbocyclic group; R<SP>2</SP>is an oxygen functional group, a nitrogen functional group, a hydrogen atom, a halogen atom, a trifluoromethyl group or -(OC<SB>2</SB>H<SB>4</SB>)<SB>n</SB>-OCH<SB>3</SB>(n is 1-4); R<SP>3</SP>is -N(R<SP>31</SP>)(R<SP>32</SP>) (R<SP>31</SP>and R<SP>32</SP>are each a hydrogen atom or an alkyl group); and X is a nitrogen atom or a carbon atom). <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a novel fluorescent molecule having a so-called dual wavelength property that identifies a specific metal and exhibits new fluorescence by addition of the metal, and a method of using the same.

Analysis of metal ions, including alkali metals and transition metals, is diverse, including industrial and living environment analysis such as rivers and seawater, composition analysis of various industrial products, blood tests, and dynamic analysis of metal ions in vivo. One of the important technologies in the field. Methods for analyzing metal ions include emission spectroscopy, atomic absorption, electrochemical analysis, optical analysis, and chromatographic analysis, each of which has its own characteristics. Examples of a simple analysis method using an instrument include an electrochemical analysis method and an optical analysis method. The most frequently used optical analysis method is the fluorescence method. The fluorescence method basically analyzes a metal ion using a substance (fluorescence probe) that emits fluorescence by forming a complex with the metal ion, and has high sensitivity and high metal ion selectivity. Things are sought.

Currently, various quinoline compounds are known. In particular, 8-hydroxyquinoline compounds form a stable complex with metal ions, and thus are widely used as reagents for detecting metal ions using these compounds. . In addition, since many metal complexes obtained by the reaction of 8-hydroxyquinoline-based compounds and metal ions exhibit fluorescence, applications to fluorescent dyes using these are considered. However, it cannot be said that these fluorescent dyes still have sufficient characteristics. For example, metal complexes such as tris (8-quinolite) aluminum are currently chemically unstable during electroluminescence and have problems such as deterioration due to short-time emission.

In such a background, research on fluorescent molecules for identifying biologically relevant substances and visualizing the dynamics of metals in the living body has been actively conducted in recent years. Many fluorescent molecules known so far are those that emit fluorescence for the first time when a metal ion binds to them, or those that emit fluorescence when the compound itself fluoresces and its fluorescence intensity increases by binding to a metal. However, high-sensitivity fluorescent molecules that have the property of identifying a specific metal and exhibiting new fluorescence by the addition of the metal (hereinafter referred to as “dual wavelength”) have the advantage that the concentration of the target compound can be measured. ,Attention has been paid. In particular, it has become clear that zinc ions play an important role in vivo, but for detailed elucidation, they can selectively bind to zinc and measure its dynamics and have dual wavelength properties. It is important to develop fluorescent molecules with high sensitivity.

The present inventors have succeeded in synthesizing compounds in which a quinolinecarboxylic acid is used as a mother nucleus, an oxygen functional group is introduced at the 4-position, and a dimethylamino group is introduced at the 6-position, and these compounds were announced at the 2003 Pharmaceutical Society. (See Non-Patent Document 1). When these compounds coexist with zinc ions, the fluorescence at 422 nm decreases depending on the concentration of zinc ions, and has a dual wavelength property that emits new fluorescence at 503 nm. However, these compounds exhibit such dual wavelength properties only in non-polar solvents such as toluene, but hardly exhibit such behavior in polar solvents, and are inferior in metal coordination ability and selectivity. There were enough points, and there were problems in actual use. Further, as a compound having a similar function, Non-Patent Document 2 (J. Am. Chem. Soc.
2002, 124 , 10650-10651), Non-Patent Document 3 (J. Am. Chem. Soc. 2003, 125 , 11458-11459), Non-Patent Document 4 (J. Am. Chem. Soc.
2004, 126 , 712-713), etc. have been reported, but the shift in wavelength before and after the coordination of zinc is small, the background is high due to factors other than zinc, etc. There were various problems. For this reason, there has been a strong demand for the development of a completely new fluorescent molecule that improves the coordination ability and selectivity for metals, and exhibits a dual wavelength with high sensitivity even in polar solvents.

Specific examples of fluorescent molecules known so far are as follows.

The following compounds developed by the present inventors have dual wavelength properties as described above. However, this effect is only seen in nonpolar solvents. Further, structurally, it has a quinolinecarboxylic acid as a mother nucleus and has an oxygen functional group at the 4-position and a dimethylamino group at the 6-position.

Non-Patent Document 2 (J. Am. Chem. Soc. 2002, 124 , 10650-10651) reports the following compounds in which zinc ions are selectively bound and exhibit dual wavelength properties. It does not have a quinoline skeleton or a naphthalene skeleton.

Non-Patent Document 3 (J. Am. Chem. Soc. 2004, 126 , 712-713) reports the following compounds that selectively bind zinc ions and exhibit dual wavelength properties. It does not have a quinoline skeleton or a naphthalene skeleton.

Non-Patent Document 4 (J. Am. Chem. Soc. 2003, 125 , 11458-11459) reports the following compounds in which zinc ions are selectively bound and exhibit dual wavelength properties. The quinoline skeleton or naphthalene skeleton is not a basic skeleton.

Patent Document 1 (Japanese Patent Laid-Open No. 11-199581) discloses the following fluorescent light-emitting cyclic phenol sulfide derivative. This substance has a fluorescence intensity near 510 nm reduced by the presence of Al ions. It does not show dual wavelength. Further, it is not a compound having a quinoline skeleton and a naphthalene skeleton.

Patent Document 2 (Japanese Patent Application Laid-Open No. 2004-317398) discloses “having a quinoline backbone structure” as one of fluorescent labeling agents for biomolecules. Biomolecules target nucleic acids, proteins, and sugar chains, and do not describe metal ions.

Patent Document 3 (Japanese Patent Laid-Open No. 2004-196683) discloses a method for measuring the fluorescence intensity of a quinoline oxide derivative (the following formula) that oxidizes melatonin and emits fluorescence in order to measure the melatonin content as an in vivo substance. The quinoline oxide derivative is shown. The excitation spectrum is 245 nm and the fluorescence spectrum is 376-8 nm. There is no description about the measurement of the metal concentration or about the dual wavelength property.

Patent Document 4 (Japanese Patent Application Laid-Open No. 2002-138081) discloses a novel quinoline compound (the following formula) suitable as a raw material for a metal complex useful as a fluorescent dye or a material for an organic EL device. Examples of metals that can form complexes with quinoline compounds include Li, Na, K, Rb, Be, Mg, Ca, Sr, Ba, Zn, Al, Ga, In, Sc, Y, and rare earth metals. Zn is listed in an enumeration but is not shown in the examples. Furthermore, there is no description of dual wavelength properties that show different fluorescence wavelengths by forming a complex.

［式中、Ｒ¹およびＲ²は、水素原子、アルキル基、アリール基、芳香族複素環基、またはシアノ基を表わすが、同時に水素原子となることはない。］
また、本発明は、一般式［１］において、Ｒ¹またはＲ²が、アリール基であることを特徴とする上記キノリン
系化合物に関する。また、本発明は、２−メチル−５−アリール−８−ヒドロキシキノリン
である上記キノリン 系化合物に関する

[Wherein, R ¹ and R ² represent a hydrogen atom, an alkyl group, an aryl group, an aromatic heterocyclic group, or a cyano group, but are not simultaneously hydrogen atoms. ]
The present invention also relates to the above quinoline compound, wherein in the general formula [1], R ¹ or R ² is an aryl group. The present invention also relates to the above quinoline compound which is 2-methyl-5-aryl-8-hydroxyquinoline.

Patent Document 5 (Japanese Patent Laid-Open No. 10-226691) discloses a quinoline compound represented by the following formula as a dye useful for a light emitting layer, a fluorescent material and an ultraviolet absorbing material of an electroluminescence element. However, there is no description about dual wavelength characteristics.

Indo 1, a doujin chemical product represented by the following formula, has the same excitation wavelength at around 330 nm for both Ca ²⁺ Free and Ca ²⁺ complexes, but the fluorescence wavelengths are 485 nm and 410 nm, respectively. And have different features. An increasing number of reports use Indo 1 to measure intracellular Ca ²⁺ trends by flow cytometry. However, this compound does not have a quinoline skeleton or a naphthalene skeleton (see, for example, Non-Patent Document 5).

Fura-2, an intracellular calcium ion measuring reagent represented by the following formula, is a fluorescent molecule that can measure Ca ²⁺ concentration up to around 1 μmol / l. The peak of the excitation wavelength is markedly blue shifted (362 nm → 335 nm) due to Ca ²⁺ binding, whereas the fluorescence intensity increases with increasing Ca ²⁺ concentration when excited near 335 nm. Conversely, when excited at around 370 to 380 nm, the fluorescence intensity decreases. Therefore, when appropriate two wavelengths are selected and excited, and the ratio of the fluorescence intensity at that time is taken, it can be correlated with the Ca ²⁺ concentration regardless of the concentration of the dye, the intensity of the light source, the size of the cell, and the like. The fluorescence wavelength does not change due to the binding of Ca ²⁺ (for example, see Non-Patent Document 6).

Quin 2 is an intracellular calcium ion measuring reagent that is a doujin chemical product represented by the following formula, and is a fluorescent indicator specific to calcium synthesized by RY Tsien et al., And exhibits strong fluorescence when complexed with calcium. Reaction with magnesium is negligible. By incorporating a fluorescent quinoline skeleton into the molecular structure of calcium-selective GEDTA, it was possible to select about 10 ⁵ units with respect to Mg ²⁺ (logK _Ca = 7.1, logK _Mg = 2.7). Due to the presence of the methoxy group, high fluorescence quantum yield is exhibited (Ca complex φ = 0.14). The calcium complex has a fluorescence excitation wavelength of 339 nm and a fluorescence wavelength of 492 nm, and does not exhibit a so-called dual wavelength property (see, for example, Non-Patent Document 7).

JP-A-11-199581 JP 2004-317398 A Japanese Patent Laid-Open No. 2004-196683 JP 2002-138081 A Japanese Patent Laid-Open No. 10-226691 Abstracts of Annual Meeting of the Pharmaceutical Society of Japan 30 [P1] I-126 Satoko Maruyama, Kazuya Kikuchi, Tomoya Hirano, Yasuteru Urano, and Tetsuo Nagano, A Novel, Cell-Permeable, Fluorescent Probe for Ratiometric Imaging of Zinc Ion. J. Am. Chem. Soc., 124, 10650-10651 (2002) Masayasu Taki, Janet L. Wolford, and Thomas V. O'Halloran, EmissionRatiometric Imaging of Intracellular Zinc: Design of a Benzoxazole FluorescentSensor and Its Application in Two-Photon Microscopy.J. Am. Chem. Soc., 126, 712-713 (2004) Carolyn C. Woodroofe and Stephen J. Lippard, A Novel Two-Fluorophore Approach to Ratiometric Sensing of Zn2 + .J. Am. Chem. Soc., 125, 11458-11459 (2003) E. M, U. Panne, Reinhard Niessner, A Fiber Optic Sensor Array for the Fluorimetric Detection of Heavy Metals, Anal.Methods Instrum., 2 (4), 182 (1995). Hiroshi Ozaki, Koichi Sato, Takumi Satoh, Hideaki Karaki, Simultaneous Recordings of Calcium Signals and Mechanical Activity Using Fluorescent Dye Fura 2 in Isolated Strips of Vascular Smooth Muscle, Jpn. J. Pharmacol., 45, 429 (1987). E. Muller-Ackermann, U. Panne, R. Niessner, A Fiber Optic SensorArray for the Fluorimetric Detection of Heavy Metals, Anal.Methods Instrum., 2 (4), 182 (1995).

In the analysis of metal ions by the fluorescence method, it has a completely new structure that can overcome the above-mentioned problems, can identify a specific metal, and has a dual wavelength property that shows a new fluorescence by adding the metal It is to provide a novel fluorescent molecule and a method for using the same.

In order to solve the above-mentioned problems, the present inventors have conducted extensive research and as a result, completed the present invention. That is, the present invention is as follows.
(1) A quinoline compound or naphthalene compound represented by the following general formula <I>.
General formula <1>

［式中、Ｒ^１は、窒素原子、イオウ原子及び酸素原子から選ばれるヘテロ原子を１乃至４個含む５員乃至６員の複素環基、又は―Ｃ_３−１０炭素環基を表し、これら複素環基および炭素環基は、オキソ基、水酸基またはＣ_１−６アルコキシ基から選ばれる酸素官能基によって置換されてもよい。Ｒ^２は、オキソ基、水酸基またはＣ_１−６アルコキシ基から選ばれる酸素官能基、アミノ基、Ｃ_１−６アルキルアミノ基、Ｃ_１−６ジアルキルアミノ基から選ばれる窒素官能基、水素原子、ハロゲン原子、トリフルオロメチル基、又は―（ＯＣ_２Ｈ_４）n−ＯＣＨ_３（ここでn＝１〜４）を表す。Ｒ^３は―Ｎ（Ｒ^３１）（Ｒ^３２)（ここでＲ^３１ とＲ^３２は、同一又は異なってもよく、水素原子または―Ｃ_１−６アルキル基を示し、Ｒ^３１
とＲ^３２は、隣接する窒素原子と一緒になって、４乃至７員の複素環を形成してもよい。）を表す。Ｘは窒素原子または炭素原子を表す。］
（２）Ｒ^１が上記酸素官能基で置換されてもよいフェニル基、ナフチル基又はチエニル基であり、Ｒ^２がハロゲン原子、Ｃ_１−６アルコキシ基又は―（ＯＣ_２Ｈ_４）nーＯＣＨ_３（ここでn＝１〜２）であり、Ｒ^３がピペラジノ基又はピペリジノ基である（１）に記載のキノリン化合物又はナフタレン化合物。
（３）（１）又は（２）に記載のキノリン化合物又はナフタレン化合物のうち少なくとも１種を含むことを特徴とする金属イオンセンシング剤。
（４）（３）に記載の金属イオンセンシング剤を用いて金属イオン濃度を測定することを特徴とする金属イオン分析方法。
（５）金属イオンが１２族金属イオン（亜鉛、カドミウム、水銀）である（４）に記載の金属イオン分析方法。

[Wherein, R ¹ represents a 5- to 6-membered heterocyclic group containing 1 to 4 heteroatoms selected from a nitrogen atom, a sulfur atom and an oxygen atom, or a —C _3-10 carbocyclic group, and The heterocyclic group and carbocyclic group may be substituted with an oxygen functional group selected from an oxo group, a hydroxyl group, or a C _1-6 alkoxy group. R ² represents an oxygen functional group selected from an oxo group, a hydroxyl group or a C _1-6 alkoxy group, an amino group, a C _1-6 alkylamino group, a nitrogen functional group selected from a C _1-6 dialkylamino group, a hydrogen atom, A halogen atom, a trifluoromethyl group, or — (OC ₂ H ₄ ) n —OCH ₃ (where n = 1 to 4) is represented. R ³ is ^{-N (R 31) (R 32} ) ( wherein ^{R 31} and ^{R 32} may be the same or different, a hydrogen atom or a _{-C 1-6} alkyl ^{group, R 31}
And R ³² together with the adjacent nitrogen atom may form a 4- to 7-membered heterocyclic ring. ). X represents a nitrogen atom or a carbon atom. ]
(2) R ¹ is a phenyl group, a naphthyl group, or a thienyl group that may be substituted with the oxygen functional group, and R ² is a halogen atom, a C _1-6 alkoxy group, or — (OC ₂ H ₄ ) n —OCH. ₃ (where n = 1 to 2), and R ³ is a piperazino group or piperidino group. The quinoline compound or naphthalene compound according to (1).
(3) A metal ion sensing agent comprising at least one of the quinoline compound or naphthalene compound according to (1) or (2).
(4) A metal ion analysis method, wherein the metal ion concentration is measured using the metal ion sensing agent according to (3).
(5) The metal ion analysis method according to (4), wherein the metal ion is a group 12 metal ion (zinc, cadmium, mercury).

The novel fluorescent molecules of the present invention, for example, in both polar and nonpolar solvents, depending on the concentration of Zn ²⁺ , for example, the fluorescence at 422 nm decreases and new fluorescence appears at 503 nm. It proved to be useful for the measurement of ²⁺ concentration. In this way, it exhibits dual wavelength not only in nonpolar solvents but also in polar solvents (methanol, hydrous dioxane, water, HEPES buffer, etc.) as a solvent, so it is possible to measure a wide range of metal ion concentrations in various solvents. It is possible and convenient. It was also revealed that strong fluorescence was emitted at sites where metal ions were localized by acting on a biological tissue slice as well as in a solvent. Furthermore, since the difference between the newly appearing fluorescence wavelength and the original fluorescence wavelength is as large as about 80 nm, the metal ion concentration can be measured correctly. It was also revealed that Ni ²⁺ and Cu ²⁺ did not show such dual wavelength properties and had high metal ion discrimination.

The definition of each substituent used in the present specification is as follows.
The “C _1-6 alkyl group” is a linear or branched alkyl group having 1 to 6 carbon atoms, such as a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, or an isobutyl group. , Sec-butyl group, tert-butyl group, pentyl group, isopentyl group, neopentyl group, tert-pentyl group, hexyl group, etc., preferably a linear or branched alkyl having 1 to 4 carbon atoms It is a group. Particularly preferred is a methyl group, an ethyl group, or an isopropyl group.

The “C _1-6 alkylamino group” is a group in which the “C _1-6 alkyl group” is bonded to an amino group, such as methylamino, ethylamino, n-propylamino, isopropylamino, n-butyl. Amino, isobutylamino, s-butylamino, tert-butylamino, n-pentylamino, isopentylamino, 2-methylbutylamino, neopentylamino, 1-ethylpropylamino, n-hexylamino, isohexylamino, 4 -Methylpentylamino, 3-methylpentylamino, 2-methylpentylamino, 1-methylpentylamino, 3,3-dimethylbutylamino, 2,2-dimethylbutylamino, 1,1-dimethylbutylamino, 1,2 -Dimethylbutylamino, 1,3-dimethylbutylamino, 2,3-dimethyl Chiruamino, and 2-ethyl-butylamino group. R ² is preferably a C _1-4 alkylamino group such as methylamino, ethylamino, n-propylamino, or isopropylamino group.

The “C _1-6 dialkylamino group” is a group obtained by substituting two identical or different “C _1-6 alkyl groups” with an amino group, for example, N, N-dimethylamino, N, N— Diethylamino, N, N-di-n-propylamino, N, N-diisopropylamino, N, N-di-n-butylamino, N, N-diisobutylamino, N, N-di-s-butylamino, N, N- Di-tert-butylamino, N, N-di-n-pentylamino, N, N-diisopentylamino, N, N-di2-methylbutylamino, N, N-dineopentylamino, N, N-di 1-ethylpropylamino, N, N-di-n-hexylamino, N, N-diisohexylamino, N, N-di4-methylpentylamino, N, N-di3-methylpentylamino, N, N -Di-2-methyl pliers Amino, N, N- di 1-methylpentylamino, N, N- ethylmethylamino, N, may be mentioned N- isopropyl-methyl-amino group. R ² is preferably a C _1-4 dialkylamino group such as N, N-dimethylamino, N, N-diethylamino, N, N-di-n-propylamino, or N, N-diisopropylamino group.

The “halogen atom” is a chlorine atom, a bromine atom, a fluorine atom or the like. R ³ is preferably a fluorine atom.

The “C _1-6 alkoxy group” is an alkyl-oxy group whose alkyl moiety is the above-defined “C _1-6 alkyl group”, specifically, a methoxy group, an ethoxy group, a propoxy group, an isopropyloxy group. Group, butoxy group, isobutyloxy group, tert-butyloxy group, pentyloxy group, hexyloxy group and the like. R ¹ is preferably a C _1-6 alkoxy group such as a methoxy group, an ethoxy group, a propoxy group, or an isopropyloxy group.

“C _3-10 carbocyclic group” is a saturated or unsaturated cyclic hydrocarbon group having 3 to 10 carbon atoms, and means an aryl group, a cycloalkyl group, a cycloalkenyl group, or a condensed ring thereof. An aryl group is preferable. Specific examples of the aryl group include a phenyl group, a naphthyl group, a pentarenyl group, and an azulenyl group, preferably a phenyl group and a naphthyl group, and particularly preferably a phenyl group. Specific examples of the “cycloalkyl group” include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, a cyclooctyl group, an adamantyl group, a norbornanyl group, etc., preferably a cyclopropyl group, a cyclobutyl group , C _3-6 cycloalkyl groups such as a cyclopentyl group and a cyclohexyl group. The “cycloalkenyl group” includes at least one, preferably 1 or 2, double bonds, specifically, a cyclopropenyl group, a cyclobutenyl group, a cyclopentenyl group, a cyclopentadienyl group, a cyclohexenyl group, Examples thereof include a cyclohexadienyl group (2,4-cyclohexadien-1-yl group, 2,5-cyclohexadien-1-yl group, etc.), cycloheptenyl group, cyclooctenyl group and the like. Specific examples of the ring in which these “aryl group”, “cycloalkyl group” and “cycloalkenyl group” are condensed include indenyl group, indanyl group, 1,4-dihydronaphthyl group, 1,2,3,4-tetrahydro Examples thereof include a naphthyl group (1,2,3,4-tetrahydro-2-naphthyl group, 5,6,7,8-tetrahydro-2-naphthyl group, etc.), a perhydronaphthyl group, and the like. Preferred is a condensed ring of a phenyl group and another ring, such as an indenyl group, an indanyl group, a 1,4-dihydronaphthyl group, a 1,2,3,4-tetrahydronaphthyl group, and particularly preferably an indanyl group.

Next, various substituents or substitution positions will be described in more detail as follows.

Examples of the “5- to 6-membered aromatic heterocyclic group containing 1 or 2 heteroatoms selected from a nitrogen atom, a sulfur atom and an oxygen atom” in R ¹ include furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, Isothiazolyl, imidazolyl, pyrazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, furazanyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1, 3,4-thiadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, tetrazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc. 5- to 6-membered aromatic monocyclic heterocyclic groups, etc. Can be mentioned.

Examples of the “4- to 7-membered saturated heterocyclic ring formed together with the adjacent nitrogen atom” in R ³ include a piperazino group and a piperidino group.

R ¹ is preferably a pyridyl group, a phenyl group, or a thienyl group, which may be substituted with an oxygen functional group selected from an oxo group, a hydroxyl group, or a C _1-6 alkoxy group, and more preferably a methoxy group. It is a pyridyl group which may be substituted, and most preferably a pyridyl group. R ² is preferably a hydrogen atom, a halogen atom, a methoxy group, or — (OC ₂ H ₄ ) n —OCH ₃ (where n = 2), more preferably a hydrogen atom, a fluorine atom, or a methoxy group. is there. The substitution position is preferably the 8-position of the quinoline ring. R ³ is preferably a dimethylamino group or a piperazino group, and more preferably a piperazino group. X is preferably a nitrogen atom.

Preferred specific examples of the quinoline compound or naphthalene compound of the present invention are listed below:
Compound (a): Pyridin-2-yl-6-pyrrolidine-1-ylquinoline,
Compound (b): 8-fluoro-2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline,
Compound (c): 8-methoxy-2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline,
Compound (d): 8- [2- (2-methoxyethoxy) -ethoxy] -2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline,
Compound (e): 2-phenyl-6-pyrrolidin-1-ylquinoline,
Compound (f): 6-pyrrolidin-1-yl-2-thiophen-2-ylquinoline,
Compound (g): 2- (4-methoxypyridin-2-yl) -6-pyrrolidin-1-ylquinoline

Among the compounds listed above, the following compounds are particularly preferable.
Compound (c): 8-methoxy-2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline,
Compound (d): 8- [2- (2-methoxyethoxy) -ethoxy] -2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline,

  The metal ion sensing agent of this invention may contain only 1 type of the quinoline compound or naphthalene compound represented by general formula <I>, or may contain 2 or more types. The metal ion sensing agent of the present invention may be a solution by dissolving the quinoline compound or naphthalene compound represented by the general formula <I> in a solvent. In the method for analyzing metal ions using the metal ion sensing agent of the present invention, a solution in which a metal is dissolved is added to a solution in which one or more quinoline compounds or naphthalene compounds represented by the general formula <I> are dissolved. Then, by measuring and comparing the fluorescence intensity of a specific wavelength before and after the addition, the metal ion concentration can be known. As a solvent for dissolving the quinoline compound or naphthalene compound represented by the general formula <I>, a nonpolar solvent such as toluene and a polar solvent such as methanol or hydrous dioxane can be used.

As these solvents, a solvent that dissolves the fluorescent molecule represented by the general formula <I> and does not affect the fluorescence measurement is used. Suitable examples include toluene, methanol, hydrous dioxane, water, HEPES buffer, DMSO and the like.

In addition, the metal ion analysis method of the present invention can be applied to a biological tissue section or the like. Thereby, the localization site | part and quantity of the metal ion in a tissue section can be measured.

Examples of metals that can be analyzed by the metal ion sensing agent of the present invention include group 12 metals, and zinc and cadmium are particularly suitable.

The metal ion concentration is not particularly limited, and is usually 1 × 10 ⁻⁶ M or more, and may be present in a large excess as compared with the quinoline compound or naphthalene compound of the present invention.

Next, although an example of the manufacturing method of the quinoline compound or naphthalene compound represented by general formula <I> is demonstrated, the manufacturing method of this invention is not limited to this. Moreover, when performing the reaction mentioned later, functional groups other than the site may be protected in advance if necessary, and this may be deprotected at an appropriate stage. Furthermore, in each step, the reaction may be carried out by a usual method, and isolation and purification are carried out by appropriately selecting or combining conventional methods such as crystallization, recrystallization, column chromatography and preparative HPLC. Just do it.

For general preparation method of manufacturing the general formula <1> a compound represented by (X = N) illustrate an example below.

Process 1

Process 2

(Wherein R1, R2 and R3 are the same as above)

Process 1
Under an argon atmosphere at 0 ° C. to room temperature, preferably at 0 ° C., a methanol solution of benzyltrimethylammonium hydroxide is added to an anhydrous THF solution of 2-acetylpyridine, and an anhydrous THF solution of compound [1] is further added, Stir at 0 ° C. for 1 to 2 hours and further at room temperature for 10 to 25 hours. The reaction mixture is extracted with ethyl acetate, the organic layer is dried over anhydrous magnesium sulfate and filtered, and the filtrate is concentrated under reduced pressure. The residue can be purified by normal phase silica gel column chromatography (methanol: water = 9: 1) to give compound [2].

Process 2
Under an argon atmosphere, an anhydrous toluene solution of R ³ -H compound such as pyrrolidine and sodium-tert-butoxide is added to an anhydrous toluene suspension of the above compound [2], palladium (II) acetate, sodium-tert-butoxide for 2 to 5 hours. Stir. To the reaction mixture are further added palladium (II) acetate and an anhydrous toluene solution of sodium-tert-butoxide, and the mixture is stirred at room temperature for about 30 minutes and at 50 ° C. for several hours. The reaction mixture is diluted with ethyl acetate and filtered, and the filtrate is concentrated under reduced pressure. The residue is purified by reverse phase silica gel column chromatography (methanol: water = 4: 1), and further purified by normal phase flash silica gel column chromatography (hexane: ether = 3: 1) to obtain compound [3]. Can do.

The above method is a method of forming a quinoline ring using compound [1] as a starting material, but is not limited to the above method, and R ¹ , R ² , R ³ groups are appropriately introduced into known quinoline derivatives according to known methods. May be. Even if the compound [1] is not known, since the compound [1] is a simple compound, those skilled in the art will be able to appropriately synthesize it by a known synthesis method. A compound in which X is a carbon atom can be produced by introducing R ¹ , R ² , and R ³ groups into a known naphthalene derivative according to a known method.

EXAMPLES Hereinafter, although an Example demonstrates this invention further in detail, this invention is not restrict | limited to these Examples.

Production of compound (a)

Production of 6-bromo-2-pyridin-2-ylquinoline (compound 2) :

Under an argon atmosphere, 40% benzyltrimethylammonium hydroxide in methanol (727 mL, 1.6 mmol) was added to a solution of 2-acetylpyridine (44.8 mL, 0.40 mmol) in anhydrous THF (4.0 mL) at 0 &ordm; C. Then, a solution of 2-amino-5-bromobenzaldehyde (88.0 mg, 0.44 mmol) in anhydrous THF (0.5 mL) was added, and the mixture was stirred at the same temperature for 1 hour. Furthermore, after stirring at room temperature for 20 hours, a saturated aqueous ammonium chloride solution (4.0 mL) was added and stirred for 5 minutes. The reaction mixture was extracted three times with ethyl acetate, the organic layer was dried over anhydrous magnesium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by normal phase silica gel column chromatography (hexane: ethyl acetate = 5: 1) to give compound 2 as white crystals (115.9 mg, 0.40 mmol, 100% yield).

White crystal mp 122-123 &ordm; C.IR (KBr) ncm ^-1 ; 1595, 1487, 1432, 785. ¹ H NMR (300 MHz,
CD ₃ OD) d; 7.52 (1H, ddd, J = 1.0, 4.8, 7.5 Hz), 7.89 (1H, dd, J =
2.2, 9.1 Hz), 8.02 (1H, ddd, J = 1.8, 7.5, 8.0 Hz), 8.06 (1H, d, J =
9.1 Hz), 8.18 (1H, d, J = 2.2 Hz), 8.38 (1H, d, J = 8.7 Hz), 8.52
(1H, d, J = 8.7 Hz), 8.59 (1H, dt, J = 1.0, 8.0 Hz), 8.73 (1H,
ddd, J = 1.0, 1.8, 4.8 Hz). 13 C NMR (75 MHz, CD 3 OD)
d; 120.9, 121.8, 123.2, 125.8, 130.8, 131.0, 132.3, 134.4, 137.5,
138.7, 147.7, 150.3, 157.0, 157.7.

Production of 2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline (compound (a)):

Under argon atmosphere, Compound 2 (122.0 mg, 0.43 mmol), Palladium (II) acetate (3.84 mg, 17.1
mmol), sodium-tert-butoxide (53.5 mg, 0.56 mmol) in anhydrous toluene (0.6 mL) suspension, pyrrolidine (47.1 mL, 0.56 mmol), tri-tert-butylphosphine (3.84 mL, 15.4 mmol) Solution of anhydrous toluene (0.1 mL) was added and stirred at room temperature for 3 hours. Further palladium (II) acetate (3.84
mg, 17.1 mmol), sodium-tert-butoxide (53.5 mg, 0.56 mmol), pyrrolidine (47.1 mL, 0.56 mmol), tri-tert-butylphosphine (3.84 mL, 15.4 mmol) in anhydrous toluene (0.1 mL) And stirred at room temperature for 30 minutes and 50 &ordm; C for 1 hour. After cooling to room temperature, saturated brine (3 mL) and ethyl acetate (3 mL) were added to the reaction mixture, and the mixture was stirred for 5 min. After separating the organic layer, the aqueous layer was extracted twice with ethyl acetate. The obtained organic layers were combined, dried over anhydrous magnesium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by normal phase silica gel column chromatography (hexane: ethyl acetate = 10: 1) to obtain yellow crystals (89.4 mg, 0.32 mmol, 76% yield) of compound (a).

Yellow crystal mp 185-188 &ordm; C. IR (KBr) ncm ^-1 ; 1622, 1590, 1509, 1390,
813. ¹ H NMR (300 MHz, C ₆ D ₆ ) d; 1.50-1.67
(4H, m), 2.95-3.10 (4H, m), 6.60 (1H, d, J = 2.7 Hz), 6.85 (1H, ddd, J
= 1.2, 4.8, 7.4 Hz), 7.03 (1H, dd, J = 2.7, 9.2 Hz), 7.42 (1H, ddd, J
= 1.8, 7.4, 8.0 Hz), 7.94 (1H, d, J = 8.7 Hz), 8.45 (1H, d, J =
9.2 Hz), 8.74 (1H, ddd, J = 0.9, 1.8, 4.8 Hz), 9.13 (2H, d, J =
^{. 8.7 Hz) 13 C NMR (} 75 MHz, C 6 D 6) d; 25.4,
47.5, 104.1, 119.2, 119.7, 121.1, 123.1, 130.8, 131.2, 134.3, 136.4, 142.4,
146.3, 149.2, 152.2, 157.6. HRESIMS Calcd for C ₁₈ H ₁₈ N ₃ :
276.1501 (M + H) ⁺ , Found: 276.1520. Anal. Calcd for C ₁₈ H ₁₇ N ₃ :
C, 78.40; H, 6.23; N, 15.60. Found: C, 78.52; H, 6.22; N, 15.26.

Production of compound (b)

Preparation of 2-fluorophenyl-carbamic acid-tert-butyl ester (compound 4) :

Di-tert-butyl dicarbonate (8.06 mL, 35.1 mmol) was added to a solution of 2-fluoroaniline (2.61 mL, 27.0 mmol) in anhydrous 1,4-dioxane (5 mL) under an argon atmosphere, and at room temperature for 4 hours. The mixture was stirred at 50 &ordm; C for 48 hours, and concentrated under reduced pressure. The residue was purified by normal phase silica gel column chromatography (hexane: ethyl acetate = 20: 1) to give compound 4 as white crystals (5.70 mg, 27.0 mmol, 100% yield).

White crystal mp 44-45 &ordm; C. IR (KBr) ncm ^-1 ; 3300, 2983, 1708, 1525,
1260, 1158, 754. ¹ H NMR (300 MHz, CDCl ₃ ) d; 1.53 (9H,
s), 6.72 (1H, bs ), 6.91-7.13 (3H, m), 8.08 (1H, t, J = 7.9 Hz). 13 C
NMR (75 MHz, C ₆ D ₆ ) d; 28.2, 80.9, 114.6 (d,
J = 19.2 Hz), 120.0 (d, J = 7.1 Hz), 122.8 (dd, J = 2.0,
7.4 Hz), 124.4 (d, J = 3.6 Hz), 126.8 (dd, J = 6.0, 10.0 Hz),
152.0 (dd, J = 4.8, 242.1 Hz), 152.3 (d, J = 4.5 Hz).

Preparation of 2-fluoro-6-formylphenyl-carbamic acid-tert-butyl ester (compound 5) :

Under an argon atmosphere, tert-BuLi (1.47 M pentane solution, 90.7 mL, 133.3 mmol) was added dropwise to a solution of compound 4 (12.8 g, 60.6 mmol) in anhydrous THF (90.0 mL) at -78 &ordm; C and stirred for 1 hour. Then, the mixture was further stirred at −20 &ordm; C for 3 hours. Anhydrous DMF (10.3 mL, 133.3 mmol) was added dropwise to the reaction mixture, and the mixture was stirred at the same temperature for 30 minutes. A saturated aqueous ammonium chloride solution (150 mL) was added, and the mixture was stirred at room temperature for 20 minutes. After separating the organic layer, the aqueous layer was extracted twice with ethyl acetate. The obtained organic layers were combined, dried over anhydrous magnesium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by normal phase silica gel column chromatography (hexane: ethyl acetate = 10: 1) to obtain white crystals of compound 5 (9.12 g, 38.1 mmol, 63% yield).

White crystal mp 85-86 &ordm; C. IR (KBr) ncm ^-1 ; 3277, 1720, 1702, 1528,
1268, 1157. ¹ H NMR (300 MHz, CDCl ₃ ) d; 1.49 (9H,
s), 7.22-7.38 (2H, m), 7.56 (1H, ddd, J = 0.8, 1.4, 7.5 Hz), 7.89 (1H,
bs), 9.98 (1H, d , J = 1.4 Hz). 13 C NMR (75 MHz, CDCl 3)
d; 28.0, 81.7, 121.6 (d, J = 21.0 Hz), 125.4 (d, J =
7.8 Hz), 127.4 (d, J = 12.6 Hz), 127.6 (d, J = 3.2 Hz), 129.5 (d,
J = 1.1 Hz), 153.0, 155.4 (d, J = 250.9 Hz), 191.3 (d, J =
3.1 Hz) .HRESIMS Calcd for C ₁₂ H ₁₅ FNO ₃ : 240.1036
(M + H) ⁺ , Found: 240.1021. Anal. Calcd for C ₁₂ H ₁₄ FNO ₃ :
C, 60.26; H, 6.07; N, 5.72. Found: C, 60.24; H, 5.90; N, 5.85.

Preparation of 2-fluoro-6-hydroxymethylphenyl-carbamic acid-tert-butyl ester (compound 6) :

To a solution of compound 5 (400.0 mg, 1.67 mmol) in methanol (4.0 mL) at room temperature under argon atmosphere, sodium borohydride (63.3 mg,
1.67 mmol) was gradually added and stirred at the same temperature for 15 minutes. Water was added to the reaction mixture, and the aqueous layer was extracted three times with ethyl acetate. The obtained organic layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by normal phase silica gel column chromatography (hexane: ethyl acetate = 2: 1) to give compound 6 as white crystals (398.5 mg, 1.65 mmol, 99% yield).

White crystal mp 64-66 &ordm; C. IR (KBr) ncm ^-1 ; 3327, 1711, 1512, 1468,
1251, 1161. ¹ H NMR (300 MHz, CDCl ₃ ) d; 1.48 (9H,
s), 3.72 (1H, bs), 4.53 (2H, d, J = 5.7 Hz), 6.61 (1H, bs), 6.97-7.07
(1H, m), 7.11-7.21 (2H, m). ¹³ C NMR (75 MHz, CDCl ₃ ) d; 28.1, 61.6
(d, J = 2.6 Hz), 81.3, 115.2 (d, J = 20.7 Hz), 123.4 (d, J =
12.7 Hz), 125.3, 127.2 (d, J = 8.3 Hz), 138.8, 154.8, 157.0 (d, J =
HRESIMS Calcd for C ₁₂ H ₁₇ FNO ₃ : 242.1192
(M + H) ⁺ , Found: 242.1213. Anal. Calcd for C ₁₂ H ₁₆ FNO ₃ :
C, 59.80; H, 6.52; N, 5.70. Found: C, 59.74; H, 6.68; N, 5.81.

Preparation of 2-amino-3-fluorophenyl-methanol (compound 7):

To a solution of compound 6 (8.35 g, 34.6 mmol) in chloroform (40.0 mL) at room temperature under argon atmosphere, trimethylsilyl iodide (5.62 mL,
39.5 mmol) was added and stirred at the same temperature for 30 minutes. The reaction mixture was adjusted to pH 9.0 with a saturated aqueous sodium hydrogen carbonate solution, and water was added to separate the organic layer. The aqueous layer was extracted three times with ethyl acetate, and the obtained organic layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in anhydrous THF (40 mL) at room temperature under an argon atmosphere, and tetrabutylammonium fluoride (1.0 M
THF solution, 19.8
mL, 19.8 mmol) was added, and the mixture was stirred at the same temperature for 30 minutes. Water was added to separate the organic layer, and the aqueous layer was extracted three times with ethyl acetate. The obtained organic layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure. . The residue was purified by normal phase silica gel column chromatography (hexane: ethyl acetate = 3: 1) to give compound 7 as white crystals (2.9 g, 20.5 mmol, 59% yield).

White crystals mp 70-71 &ordm; C. IR (KBr) ncm ^-1 ;
3413, 3336, 1638, 1491, 1233, 998, 778, 728. ¹ H NMR (300 MHz, CDCl ₃ )
d; 2.05 (1H, bs), 4.22 (2H, bs), 4.66 (2H, s), 6.63 (1H, dt, J =
5.1, 7.8 Hz), 6.84 (1H, d, J = 7.3 Hz), 6.96 (1H, ddd, J = 1.4,
^{. 8.2, 10.8 Hz) 13 C} NMR (75 MHz, CDCl 3) d; 63.7 (d, J
= 3.0 Hz), 115.0 (d, J = 19.2 Hz), 117.4 (d, J = 7.5 Hz),
124.1 (d, J = 2.8 Hz), 127.0 (d, J = 3.3 Hz), 134.1 (d, J =
13.0 Hz), 152.0 (d, J = 238.3 Hz). HRESIMS Calcd for C ₇ H ₉ FNO:
142.0668 (M + H) ⁺ , Found: 142.0659. Anal. Calcd for C ₇ H ₈ FNO:
C, 59.73; H, 5.84; N, 9.77. Found: C, 59.57; H, 5.71; N, 9.92.

Preparation of 2-amino-5-bromo-3-fluorophenyl-methanol (Compound 8) :

Under argon atmosphere, 0 &ordm; C was added 2,4,4,6-tetrabromo-2,5-cyclohexadienone (29.0 mg, 70.9 mmol) to a solution of compound 7 (10.0 mg, 70.9 mmol) in anhydrous dichloromethane (0.3 mL). ) Was added, and the mixture was stirred at the same temperature for 15 minutes, and further stirred at room temperature for 15 minutes. A 2N aqueous sodium hydroxide solution was added to the reaction mixture, and the organic layer was separated. The aqueous layer was extracted three times with ethyl acetate, and the obtained organic layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by normal phase silica gel column chromatography (hexane: ethyl acetate = 1: 1) to obtain white crystals of compound 8 (13.9 mg, 63.2 mmol, 89% yield).

White crystal mp 136-138 &ordm; C. IR (KBr) ncm ^-1 ; 3399, 3326, 1638, 1490,
1232, 1000, 865. ¹ H NMR (300 MHz, CDCl ₃ ) d; 1.68 (1H,
t, J = 5.5 Hz), 4.25 (2H, bs), 4.66 (2H, d, J = 5.5 Hz), 7.02
(1H, s), 7.13 ( 1H, dd, J = 2.2, 10.1 Hz). 13 C NMR (75 MHz,
CDCl ₃ ) d; 63.4 (d, J = 3.1 Hz), 107.8 (d, J = 9.5 Hz), 118.2
(d, J = 22.4 Hz), 126.9 (d, J = 3.0 Hz), 128.1 (d, J = 3.6
Hz), 133.6 (d, J = 12.9 Hz), 151.6 (d, J = 242.9 Hz). HRESIMS
Calcd for C ₇ H ₈ BrFNO: 219.9773 (M + H) ⁺ , Found:
219.9786. Anal.Calcd for C ₇ H ₇ BrFNO: C, 37.89; H, 3.58;
N, 6.00. Found: C, 38.21; H, 3.21; N, 6.37.

Preparation of 2-amino-5-bromo-3-fluorobenzaldehyde (Compound 9) :

Manganese dioxide (27.3 mg, 0.31 &#61549; mol) was added to a solution of compound 8 (23.0 mg, 0.10 &#61549; mol) in anhydrous dichloromethane (1.5 mL) at room temperature under an argon atmosphere and stirred at the same temperature for 13 hours. did. In addition, manganese dioxide (27.3 mg,
0.31 &#61549; mol) was added to the mixture and stirred for 2 hours. After filtration of the reaction mixture, the filtrate was concentrated under reduced pressure to obtain white crystals of compound 9 (21.8 mg, 0.10 &#61549; mol, 100% yield).

White crystals mp 74-76 &ordm; C. IR (KBr) ncm ^-1 ;
3444, 3337, 1669, 1629, 1551, 1475, 1230, 872.1 ¹ H NMR (300 MHz, CDCl ₃ )
d; 6.17 (2H, bs), 7.28 (1H, dd, J = 2.2, 10.5 Hz), 7.43 (1H,
t, J = 2.0 Hz), 9.81 (1H, d, J = 2.0 Hz). ¹³ C NMR (75
MHz, CDCl ₃ ) d; 105.3 (d, J = 8.8 Hz), 120.9 (d, J = 4.8 Hz), 122.6
(d, J = 21.2 Hz), 132.4 (d, J = 3.4 Hz), 138.0 (d, J =
13.7 Hz), 150.6 (d, J = 246.6 Hz), 192.2 (d, J = 2.7 Hz). HRESIMS
Calcd for C ₇ H ₆ BrFNO: 217.9617 (M + H) ⁺ , Found:
217.9635. Anal.Calcd for C ₇ H ₅ BrFNO: C, 38.79; H, 2.64;
N, 6.20. Found: C, 38.56; H, 2.31; N, 6.42.

Preparation of 6-bromo-8-fluoro-2-pyridin-2-ylquinoline (compound 10) :

A solution of potassium tert-butoxide (51.6 mg, 0.46 mmol) in tert-butanol (0.46 mL) in anhydrous THF (0.5 mL) in 2-acetylpyridine (28.1 mL, 0.25 mmol) at 0 &ordm; C in an argon atmosphere Further, Compound 9 (50.0 mg, 0.23 &#61549; mol) was added and stirred at the same temperature for 1.5 hours. After further stirring at room temperature for 30 minutes, the reaction mixture was concentrated under reduced pressure. The residue was purified by normal phase silica gel column chromatography (hexane: ethyl acetate = 5: 1) to give compound 10 as white crystals (59.1 mg, 0.17 mmol, 75% yield).

White crystal mp 155-157 &ordm; C. IR (KBr) ncm ^-1 ; 1593, 1490, 861, 777. ¹ H
NMR (300 MHz, DMSO) d; 7.68 (1H, ddd, J = 1.0, 4.8, 7.4
Hz), 8.04 (1H, dd, J = 2.0, 10.3 Hz), 8.16 (1H, ddd, J = 1.8, 7.4,
9.0 Hz), 8.32 (1H, bs), 8.68 (1H, bt, J = 9.0 Hz), 8.70 (1H, d, J =
8.7 Hz), 8.79 (1H, d, J = 8.7 Hz), 8.89 (1H, bd, J = 4.8 Hz). 13 C
NMR (75 MHz, DMSO) d; 118.7 (d, J = 22.1 Hz), 119.6 (d, J = 9.7 Hz),
121.3, 122.4, 126.1, 127.1 (d, J = 4.8 Hz), 131.2 (d, J = 2.4
Hz), 137.2 (d, J = 11.5 Hz), 137.3 (d, J = 3.0 Hz), 138.5, 150.4,
155.4, 156.9, 158.1 (d, J = 260.4 Hz) .HRESIMS Calcd for C ₁₄ H ₉ BrFN ₂ :
302.9933 (M + H) ⁺ , Found: 302.9944. Anal.Calcd for C ₁₄ H ₈ BrFN ₂ :
C, 55.22; H, 3.04; N, 9.20. Found: C, 55.47; H, 2.66; N, 9.24.

Production of 8-fluoro-2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline (compound (b)) :

Under argon atmosphere, pyrrolidine (52.3 mL, 0.62 mmol), palladium (II) acetate (1.4 mg, 6.2 mmol), (S) -BINAP (5.8 mg,
A suspension of 9.3 mmol) in anhydrous toluene (1.0 mL) was stirred at 100 &ordm; C for 3 minutes. After cooling to room temperature, compound 10 is added to the reaction mixture.
A solution of (187.5 mg, 0.62 mmol) in anhydrous toluene (1.5 mL) and sodium-tert-butoxide (83.2 mg, 0.87 mmol) were added, and the mixture was stirred at 100 &ordm; C for 1.5 hours. Furthermore, pyrrolidine (52.3 mL, 0.62 mmol), palladium (II) acetate (1.4 mg, 6.2 mmol), (S) -BINAP (5.8 mg, 9.3 mmol), sodium-tert-butoxide (83.2 mg, 0.87 mmol) were added. In addition, the mixture was stirred at 100 &ordm; C for 30 minutes. The reaction mixture was diluted with ethyl acetate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase silica gel column chromatography (methanol: water = 4: 1) and further purified by normal phase flash silica gel column chromatography (hexane: ether = 3: 1) to give yellow crystals of compound (b). (148.4 mg, 0.51 mmol, 82% yield) was obtained.

Yellow crystal mp 205-207 &ordm; C. IR (KBr) ncm ^-1 ; 1635, 1501, 1484, 1441,
774. ¹ H NMR (300 MHz, CD ₃ OD) d; 2.10-2.16 (4H, m),
3.43-3.49 (4H, m), 6.63 (1H, d, J = 2.5 Hz), 7.08 (1H, dd, J =
2.5, 13.7 Hz), 7.46 (1H, ddd, J = 1.2, 4.9, 7.5 Hz), 7.98 (1H, ddd, J
= 1.8, 7.5, 8.0 Hz), 8.17 (1H, dd, J = 1.7, 8.8 Hz), 8.33 (1H, d J
= 8.8 Hz), 8.53 (1H, bd, J = 8.0 Hz), 8.67 (1H, ddd J = 0.8,
^{. 1.8, 4.9 Hz) 13 C} NMR (75 MHz, DMSO) d; 26.1, 48.7, 100.5 (d,
J = 2.8 Hz), 104.8 (d, J = 22.2 Hz), 120.5, 121.4, 124.9, 131.4
(d, J = 11.9 Hz), 132.0 (d, J = 3.6 Hz), 135.2 (d, J = 3.3
Hz), 138.2, 147.1 (d, J = 10.6 Hz), 150.1, 151.2, 156.5, 159.0 (d, J =
HRESIMS Calcd for C ₁₈ H ₁₇ FN ₃ : 294.1407
(M + H) ⁺ , Found: 294.1422. Anal.Calcd for C ₁₈ H ₁₆ FN ₃ :
C, 73.91; H, 5.58; N, 14.28.Found: C, 73.70; H, 5.50; N, 14.32.

Preparation of 6-bromo-8-methoxy-2-pyridin-2-ylquinoline (compound 11) :

A solution of benzyltrimethylammonium hydroxide in methanol (40%, 0.30 &#61549; L, 0.66 mmol) was added to a solution of compound 10 (50.0 mg, 0.17 mmol) in anhydrous THF (0.3 mL) at room temperature under an argon atmosphere. Stir at &ordm; C for 42 hours. Water was added to the reaction mixture, and the mixture was extracted 3 times with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase silica gel column chromatography (methanol: water = 9: 1) to obtain pale yellow crystals (52.0 mg, 0.17 mmol, 100% yield) of compound 11.

Alternatively, in an argon atmosphere at 0 &ordm; C, 2-acetylpyridine (56.1 mL, 0.50 mmol) in anhydrous THF (2.0 mL) in methanol (0.4 mL) and benzyltrimethylammonium hydroxide in methanol ( 40%, 0.83 &#61549; L, 1.83 mmol), a solution of compound 9 (100.0 mg, 0.46 mmol) described in Example 2 in anhydrous THF (1.5 mL) was added and stirred for 1 hour. After further stirring at room temperature for 2.5 hours and 40 &ordm; C for 17 hours, methanol (0.5 mL) was added to the reaction mixture, followed by stirring at 60 &ordm; C for 4 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by normal phase silica gel column chromatography (hexane: ethyl acetate = 5: 1) to give pale yellow crystals of compound 11.
(116.4 mg, 0.37 mmol, 81% yield) was obtained.

Light yellow crystal mp 112-116 &ordm; C. IR (KBr) ncm ^-1 ; 1618, 1484, 1452, 1385,
1115, 778. ¹ H NMR (300 MHz, CD ₃ OD) d; 4.10 (3H,
s) 7.26 (1H, d, J = 1.9 Hz), 7.49 (1H, ddd, J = 1.2, 4.9, 7.5
Hz), 7.67 (1H, d, J = 1.9 Hz), 7.98 (1H, ddd, J = 1.7, 7.5, 8.0
Hz), 8.26 (1H, d, J = 8.7 Hz), 8.46 (1H, d, J = 8.7 Hz), 8.58
(1H, bd, J = 8.0 Hz), 8.69 (1H, ddd, J = 0.9, 1.7, 4.9 Hz). 13 C
NMR (75 MHz, CD ₃ OD) d; 56.9, 113.4, 121.4, 121.9, 122.7, 123.5,
125.6, 131.3, 137.2, 138.5, 139.7, 150.1, 156.3, 157.1, 157.4. HRESIMS Calcd
for C ₁₅ H ₁₂ BrN ₂ O: 315.0133 (M + H) ⁺ ,
Found: 315.0120. Anal. Calcd for C ₁₅ H ₁₁ BrN ₂ O:
C, 57.33; H, 3.82; N, 8.60. Found: C, 57.16; H, 3.52; N, 8.89.

Production of 8-methoxy-2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline (compound (c)) :

Under an argon atmosphere, a suspension of compound 11 (21.5 mg, 68.2 mmol), palladium (II) acetate (0.3 mg, 1.36 mmol), sodium-tert-butoxide (8.5 mg, 88.7 mmol) in anhydrous toluene (0.2 mL) was suspended. Further, a solution of pyrrolidine (7.51 mL, 88.7 mmol) in anhydrous toluene (0.05 mL) and a solution of tri-tert-butylphosphine (0.31 mL, 1.23 mmol) in anhydrous toluene (0.05 mL) were added, and the mixture was stirred at room temperature for 2.5 days. In addition, palladium (II) acetate (0.3
mg, 1.36 mmol), sodium-tert-butoxide (8.5 mg, 88.7 mmol) and pyrrolidine (7.51 mL, 88.7 mmol) in anhydrous toluene (0.05 mL), tri-tert-butylphosphine (0.31 mL, 1.23 mmol) Of anhydrous toluene (0.05 mL) was added, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by reverse phase silica gel column chromatography (methanol: water = 9: 1) to obtain yellow crystals (19.8 mg, 64.8 mmol, 95% yield) of compound (c). It was.

Yellow crystal mp 55-60 &ordm; C. IR (KBr) ncm ^-1 ; 1618, 1588, 1484, 1452,
1385, 778. ¹ H NMR (300 MHz, CD ₃ OD) d; 2.08-2.15
(4H, m), 3.42-3.50 (4H, m), 4.11 (3H, s), 6.38 (1H, d, J = 2.1 Hz), 6.69
(1H, d, J = 2.1 Hz), 7.43 (1H, ddd, J = 1.2, 4.9, 7.5 Hz), 7.96
(1H, ddd, J = 1.8, 7.5, 8.0 Hz), 8.07 (1H, d, J = 8.7 Hz), 8.23
(1H, d, J = 8.7 Hz), 8.50 (1H, ddd, J = 0.8, 1.2, 8.0 Hz), 8.64
(1H, ddd, J = 0.8, 1.8, 4.9 Hz). ¹³ C NMR (75 MHz, CD ₃ OD)
d; 26.5, 48.8, 56.4, 97.2, 100.1, 120.8, 123.1, 124.5, 132.6, 134.8,
135.6, 138.5, 148.6, 149.9, 150.7, 157.2, 158.1. HRESIMS Calcd for C ₁₉ H ₂₀ N ₃ O:
306.1606 (M + H) ⁺ , Found: 306.1578. Anal.Calcd for C ₁₉ H ₁₉ N ₃ O:
C, 74.73; H, 6.20; N, 13.40. Found: C, 74.73; H, 6.27; N, 13.76.

Preparation of 6-bromo-8- [2- (2-methoxyethoxy) -ethoxy] -2-pyridin-2-ylquinoline (Compound 12) :

2- (2-methoxyethoxy) ethanol (78.5 mL, 0.66 mmol), benzyltrimethylammonium hydroxide aqueous solution in anhydrous THF (0.2 mL) solution of compound 10 (20.0 mg, 66.0 mmol) at room temperature under argon atmosphere
(40%, 0.1 &#61549; L, 0.26 mmol) was added, and the mixture was stirred at the same temperature for 1.5 hours, and further at 50 &ordm; C for 40 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase silica gel column chromatography (methanol: water = 7: 3) to give Compound 12 as a pale yellow oil (22.4 mg, 55.5 mmol, 84% yield). Obtained.

Alternatively, in an argon atmosphere at 0 &ordm; C, 2-acetylpyridine (56.1 mL, 0.50 mmol) in 2- (2-methoxyethoxy) ethanol (1.0 mL) solution in benzyltrimethylammonium hydroxide aqueous solution (40 %, 0.72 &#61549; L, 1.83 mmol), and the compound 9 described in Example 2
(100.0 mg, 0.46 mmol) was added, and the mixture was stirred at the same temperature for 1 hour and further at room temperature for 18 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by normal phase silica gel column chromatography (hexane: ethyl acetate = 3: 1) and further purified by reverse phase silica gel column chromatography (methanol: water = 7: 3). A pale yellow oily substance of compound 12 (142.8 mg, 0.35 mmol, 77% yield) was obtained.

Pale yellow oil IR (neat) ncm ^-1 ; 2925, 2877, 1586, 1114, 782.1 ¹ H NMR (300
MHz, CD ₃ OD) d; 3.37 (3H, s), 3.58-3.64 (2H, m), 3.84-3.91 (2H, m), 3.99-4.06 (2H,
m), 4.34-4.41 (2H, m), 7.26 (1H, d, J = 1.8 Hz), 7.45 (1H, ddd, J =
1.1, 4.9, 7.5 Hz), 7.60 (1H, d, J = 1.8 Hz), 7.93 (1H, ddd, J =
1.7, 7.5, 7.9 Hz), 8.18 (1H, d, J = 8.7 Hz), 8.42 (1H, d, J = 8.7
Hz), 8.56 (1H, bd, J = 7.9 Hz), 8.66 (1H, ddd, J = 0.8, 1.7, 4.9
Hz). ¹³ C NMR (75 MHz, CD ₃ OD) d; 59.2, 70.6, 70.7,
71.9, 73.1, 115.1, 121.1, 121.8, 123.1, 123.3, 125.5, 131.4, 137.1, 138.4,
139.9, 150.1, 156.0, 156.8, 157.1. HRESIMS Calcd for C ₁₉ H ₂₀ BrN ₂ O ₃ :
403.0657 (M + H) ⁺ , Found: 403.0666. Anal. Calcd for C ₁₉ H ₁₉ BrN ₂ O ₃ :
C, 56.56; H, 4.85; N, 6.81. Found: C, 56.59; H, 4.75; N, 6.95.

Preparation of 8- [2- (2-methoxyethoxy) -ethoxy] -2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline (compound (d)) :

Under an argon atmosphere, a suspension of compound 12 (50.0 mg, 124.0 mmol), palladium (II) acetate (1.1 mg, 4.96 mmol), sodium-tert-butoxide (15.5 mg, 161.2 mmol) in anhydrous toluene (0.05 mL) was suspended. , Pyrrolidine (13.6 mL, 161.2 mmol) and tri-tert-butylphosphine (1.11 ml, 4.46 mmol) in anhydrous toluene (0.05 mL) were added, and the mixture was stirred at room temperature for 2 hours and 50 &ordm; C for 18 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by reversed-phase silica gel column chromatography (methanol: water = 1: 1) to obtain yellowish brown crystals (35.3 mg, 89.7 mmol, 72% yield) of compound (d). Obtained.

Tan crystals mp 80-85 &ordm; C. IR (KBr) ncm ^-1 ; 2874, 1610, 1440, 1388,
1103, 777. ¹ H NMR (300 MHz, CD ₃ OD) d; 2.03-2.11
(4H, m), 3.37-3.45 (4H, m), 3.38 (3H, s), 3.60-3.66 (2H, m), 3.86-3.97 (2H, m),
4.02-4.09 (2H, m), 4.40-4.46 (2H, m), 6.35 (1H, d, J = 2.3 Hz), 6.71 (1H,
d, J = 2.3 Hz), 7.41 (1H, ddd, J = 1.2, 4.9, 7.5 Hz), 7.93 (1H,
ddd, J = 1.7, 7.5, 8.1 Hz), 8.02 (1H, d, J = 8.7 Hz), 8.24 (1H,
d, J = 8.7 Hz), 8.52 (1H, bd, J = 8.1 Hz), 8.63 (1H, ddd, J =
. 0.8, 1.7, 4.9 Hz) 13 C NMR (75 MHz, CD 3 OD) d; 26.5,
48.8, 59.1, 70.4, 70.8, 71.9, 73.1, 97.7, 102.3, 120.5, 122.9, 124.5, 132.7,
135.1, 135.5, 138.4, 148.4, 149.8, 150.6, 156.6, 158.3. HRESIMS Calcd for C ₂₃ H ₂₈ N ₃ O ₃ : 394.2131
(M + H) ⁺ , Found: 394.2156.

2-Phenyl-6-pyrrolidine-1-ylquinoline
Production of (Compound (e)) The compound (e) was prepared in the same manner as in Examples 1 to 4 above.

Yellow crystal mp 174-175 &ordm; C. IR
(KBr) n cm ^-1 ; 1623, 1507, 1386, 823, 758, 691. ¹ H NMR
(300 MHz, C ₆ D ₆ ) d; 1.56-1.63 (4H, m),
3.01-3.08 (4H, m), 6.67 (1H, d, J = 2.7 Hz), 7.04 (1H, dd, J =
2.7, 9.2 Hz), 7.29-7.38 (1H, m), 7.42-7.51 (2H, m), 7.73 (1H, d, J = 8.7
Hz), 7.87 (1H, d, J = 8.7 Hz), 8.46 (1H, d, J = 9.2 Hz),
^{. 8.47-8.56 (2H, m) 13} C NMR (75 MHz, C 6 D 6) d; 25.4 (CH 2),
47.6 (CH ₂ ), 103.8 (CH), 119.0 (CH), 119.4
(CH), 127.4 (CH), 128.6 (CH), 128.8 (CH), 129.6 (C), 131.3 (CH), 134.2 (CH),
140.8 (C), 143.0 (C), 146.1 (C), 152.5 (C) .HRESIMS Calcd for C ₁₉ H ₁₉ N ₂ :
275.1548 (M + H) ⁺ , Found: 275.1569.

Production of 6-pyrrolidin-1-yl-2-thiophen-2-ylquinoline (compound (f)):
It created by the method similar to the said Examples 1-4.

Yellow crystal mp 210-213 &ordm; C. IR (KBr) n cm ^-1 ; 1619, 1501, 1387, 822,
706. ¹ H NMR (300 MHz, C ₆ D ₆ ) d; 1.52-1.62
(4H, m), 2.94-3.04 (4H, m), 6.59 (1H, d, J = 2.5 Hz), 6.94 (1H, dd, J
= 2.6, 9.2 Hz), 6.98 (1H, dd, J = 3.7, 5.0 Hz), 7.09 (1H, d, J =
5.0 Hz), 7.58 (1H, d, J = 8.6 Hz), 7.60 (1H, s), 7.74 (1H, d, J =
8.6 Hz), 8.34 (1H, d, J = 9.2 Hz). ¹³ C NMR (75 MHz, C ₆ D ₆ )
d; 25.4 (CH ₂ ), 47.5 (CH ₂ ), 104.0 (CH), 118.0 (CH), 119.4 (CH), 124.2 (CH), 127.3 (CH),
127.8 (CH), 129.5 (C), 130.7 (CH), 134.0 (CH), 142.5 (C), 145.9 (C), 147.3 (C),
148.3 (C). HRESIMS Calcd for C ₁₇ H ₁₇ N ₂ S: 281.1112
(M + H) ⁺ , Found: 281.1110.

Preparation of 2- (4-methoxypyridin-2-yl) -6-pyrrolidin-1-ylquinoline (compound (g)):
It created by the method similar to the said Examples 1-4.

Yellow crystal mp 192-195 &ordm; C. IR (KBr) n cm ^-1 ; 1617, 1594, 1565,1379,
1036, 904, 825. ¹ H NMR (300 MHz, C ₆ D ₆ ) d; 1.54-1.61
(4H, m), 2.96-3.05 (4H, m), 3.40 (3H, s), 6.62 (1H, d, J = 2.5 Hz), 6.67
(1H, dd, J = 2.6, 5.6 Hz), 7.02 (1H, dd, J = 2.6, 9.2 Hz), 7.96
(1H, d, J = 8.7 Hz), 8.46 (1H, d, J = 9.2 Hz), 8.63 (1H, d, J =
5.6 Hz), 8.79 (1H, d, J = 2.5 Hz), 9.22 (1H, d, J = 8.7 Hz). ¹³ C
NMR (75 MHz, C ₆ D ₆ ) d; 25.4 (CH ₂ ),
47.5 (CH ₂ ), 54.6 (CH ₃ ), 104.1 (CH), 105.7 (CH), 111.0
(CH), 119.2 (CH), 120.2 (CH), 130.9 (C), 131.1 (CH), 134.2 (CH), 142.4 (C),
146.3 (C), 150.4 (CH), 152.2 (C), 159.6 (C), 166.8 (C). HRESIMS Calcd for C ₁₉ H ₂₀ N ₃ O:
306.1606 (M + H) ⁺ , Found: 306.1580.

Excitation and fluorescence spectra of 8-methoxy-2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline
8-Methoxy-2-pyridin-2-yl-6-pyrrolidin-1-yl-quinoline (1 mg) is dissolved in THF, acetone, methanol, 50 mM HEPES buffer (pH 7.2) to make 10 mL. Measurements were performed using a 1000-fold diluted solution. The measurement wavelength was measured at the excitation wavelength when measuring the fluorescence spectrum, the maximum excitation wavelength near 400 nm, respectively, and the fluorescence wavelength when measuring the excitation spectrum at the maximum fluorescence wavelength. The wavelength shifted to the longer wavelength side due to the dielectric constant of the solvent (FIG. 1).

Dual wavelength in the presence of zinc in methanol.
Dissolve 8-methoxy-2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline in methanol to make 10 mL, and gradually add a methanol solution of zinc bromide dropwise to a 1000-fold diluted solution. Measured. The measurement wavelength was measured at an excitation wavelength of 330 nm. Depending on the concentration of zinc bromide, the fluorescence wavelength near 500 nm decreased and the fluorescence wavelength near 570 nm increased (FIG. 2).

Dual wavelength (1) in the presence of zinc in a buffer (50 mM HEPES buffer (pH 7.2))
8-Methoxy-2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline was dissolved in 50 mM HEPES buffer (pH 7.2) to make 10 mL, and further diluted 1000 times with 50 mM HEPES buffer of zinc chloride ( pH 7.2) The solution was gradually dropped and measured. The measurement wavelength was measured at an excitation wavelength of 433 nm. The fluorescence around 530 nm decreased and the fluorescence around 570 nm increased depending on the concentration of zinc chloride (FIG. 3).

Dual wavelength (2) in the presence of zinc in buffer (50 mM HEPES buffer (pH 7.2))
8-Methoxy-2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline (1 mg) in 50 mM HEPES buffer (pH 7.2)
To a solution diluted to 10 mL and further diluted 1000 times, a sufficient amount of a 50 mM HEPES buffer (pH 7.2) solution of zinc chloride was dropped and measured. The measurement wavelength was measured with the excitation wavelength when measuring the fluorescence spectrum being a maximum excitation wavelength around 400-500 nm, respectively, and the fluorescence wavelength when measuring the excitation spectrum being respectively the maximum fluorescence wavelength. With the addition of zinc ions, the wavelength shifted to the longer wavelength side (Fig. 4).

Metal selectivity, ratio measurement (1)
8-methoxy-2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline (1 mg) dissolved in 50 mM HEPES buffer (pH 7.2) to 10 mL, and further diluted 1000 times. Further, a 50 mM HEPES buffer (pH 7.2) solution of metal ions was added, and the transition metal was measured at 25 μM, and the alkali and alkaline earth metals were measured at 5 mM.
Excitation at 433 nm measured the ratio of fluorescence at 565 nm and 535 nm. In the presence of zinc ions and cadmium ions, the fluorescence wavelength shifted to the longer wavelength side, but no change was observed with other metal ions (FIG. 5).

Metal selectivity, ratio measurement (2)
6-Bromo-8- [2- (2-methoxyethoxy) -ethoxy] -2-pyridin-2-ylquinoline (1 mg) was dissolved in 50 mM HEPES buffer (pH 7.2) to make 10 mL, and further 1000 times A 50 mM HEPES buffer (pH 7.2) solution of the described metal ions was added to the diluted solution, and the transition metal (25 μM), alkali, and alkaline earth metal (5 mM) were measured. Excitation was performed at 433 nm and the ratio of fluorescence at 565 nm and 535 nm was measured. In the presence of zinc ion, the fluorescence wavelength shifted to the higher wavelength side, but no change was observed with other metal ions (FIG. 6).

Analysis of localization of zinc ion in brain tissue section A 300-um coronal slice of the hippocampus of a ddY mouse (6-7 weeks old) was prepared by a conventional method. After staining with a 20 uM solution of 8-methoxy-2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline for 60 minutes and washing for 30 minutes, excitation was performed at 480 nm and fluorescence at 590 nm was measured. The portion where zinc ions were localized emitted light (FIG. 7).

Analysis of zinc ion localization in brain tissue sections (ratio measurement)
A 300-um coronal slice of the hippocampus of a ddY mouse (6-7 weeks old) was prepared by a conventional method. Stained with 20 uM solution of 8-methoxy-2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline for 60 minutes, washed for 30 minutes, excited at 480 nm, and emitted fluorescence at 590 nm under fluorescence microscope-image processing system For 25 minutes. The baseline was measured for 5 minutes in the state of artificial cerebrospinal fluid, changed to ischemic Ringer (glucose-free state) for 10 minutes, then returned to artificial cerebrospinal fluid and measured for 10 minutes. The fluorescence images of each time zone were compared, and the zinc ion fluctuation of the cells on the slice was analyzed. Fluorescence microscope-Image processing system conditions are Filter1: 470-490 excitation Filter, Filter2: 400-440 excitation
Filter, dichroic mirror: DM505, fluorescence Filter: BA510 /
20, Ratio: F470-490 / F400-440. The fluctuation of zinc ion localization due to ischemia could be analyzed for each site. CA1 had the highest fluctuation rate (FIG. 8).

Excitation and fluorescence spectrum of 8-methoxy-2-pyridin-2-yl-6-pyrrolidin-1-ylquinoline: The wavelength shifted to the longer wavelength side depending on the dielectric constant of the solvent. Dual wavelength in the presence of zinc in methanol: The fluorescence wavelength near 500 nm decreased and the fluorescence wavelength near 570 nm increased depending on the concentration of zinc ions. Dual wavelength in the presence of zinc in a buffer (50 mM HEPES buffer (pH 7.2)) (1): The fluorescence wavelength near 530 nm decreased and the fluorescence wavelength near 570 nm increased depending on the concentration of zinc ions. Dual wavelength (2) in the presence of zinc in a buffer (50 mM HEPES buffer (pH 7.2)): A new fluorescence wavelength appeared on the longer wavelength side by the addition of zinc ions. Metal selectivity, ratio measurement (1): In the presence of zinc ions and cadmium ions, the fluorescence wavelength was shifted to the higher wavelength side, but no change was observed with other metal ions. Metal selectivity, ratio measurement (2): In the presence of zinc ions, the fluorescence wavelength shifted to the higher wavelength side, but no change was observed with other metal ions. Analysis of zinc ion localization in brain tissue section: The portion where zinc ions were localized emitted light. Analysis of zinc ion localization in brain tissue sections (ratio measurement): We were able to analyze the local variation of zinc ions due to ischemia.

Claims (5)

A quinoline compound or naphthalene compound represented by the following general formula <I>.
General formula <1>

[Wherein, R ¹ represents a 5- to 6-membered heterocyclic group containing 1 to 4 heteroatoms selected from a nitrogen atom, a sulfur atom and an oxygen atom, or a —C _3-10 carbocyclic group, and The heterocyclic group and carbocyclic group may be substituted with an oxygen functional group selected from an oxo group, a hydroxyl group, or a C _1-6 alkoxy group. R ² represents an oxygen functional group selected from an oxo group, a hydroxyl group or a C _1-6 alkoxy group, an amino group, a C _1-6 alkylamino group, a nitrogen functional group selected from a C _1-6 dialkylamino group, a hydrogen atom, A halogen atom, a trifluoromethyl group, or — (OC ₂ H ₄ ) n —OCH ₃ (where n = 1 to 4) is represented. R ³ is ^{-N (R 31) (R 32} ) ( wherein ^{R 31} and ^{R 32} may be the same or different, a hydrogen atom or a _{-C 1-6} alkyl ^{group, R 31}
And R ³² together with the adjacent nitrogen atom may form a 4- to 7-membered heterocyclic ring. ). X represents a nitrogen atom or a carbon atom. ] R ¹ is a phenyl group, a naphthyl group or a thienyl group which may be substituted with the oxygen functional group, and R ² is a halogen atom, a C _1-6 alkoxy group or — (OC ₂ H ₄ ) n —OCH ₃ (here N = 1-2), and R ³ is a piperazino group or a piperidino group. The quinoline compound or naphthalene compound according to claim 1. A metal ion sensing agent comprising at least one of the quinoline compound and the naphthalene compound according to claim 1. A metal ion analysis method comprising measuring a metal ion concentration using the metal ion sensing agent according to claim 3. The metal ion analysis method according to claim 4, wherein the metal ion is a group 12 metal ion (zinc, cadmium, mercury).

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