JP7264044B2 - Fluorescent Labeling Agents and Phosphorus Phthalocyanines - Google Patents

Description

The present invention relates to a fluorescent labeling agent and phosphorous phthalocyanine used therefor.

Bioimaging is a technique for visualizing proteins, cells, tissues, etc., and is widely used in the research fields of biology and medicine, such as the elucidation of in vivo molecules and cell functions and drug discovery research. Among them, the fluorescence bioimaging method is an imaging method that enables dynamic observation of phenomena, multicolor observation, and high-sensitivity observation. Furthermore, in recent years, the fluorescence bioimaging method has attracted attention as an imaging method that enables non-invasive diagnosis, and is expected to be applied in clinical settings such as image diagnosis with less burden on patients and real-time diagnosis during surgery.

Fluorescent bioimaging methods mainly use fluorescent dyes that either adsorb to the target site or emit light only at the target site, and detect the fluorescence emitted by the dye when the fluorescent dye is irradiated with light in the ultraviolet to near-infrared region. This is a method for visualizing the target.

Many dyes (fluorescent labeling agents) for fluorescent bioimaging have absorption in the visible light region, but there is a problem that the dyes are easily degraded by light irradiation because visible light has high energy. In addition, with dyes that absorb or fluoresce in the visible light region, not only the fluorescence derived from the dye, but also the fluorescence (autofluorescence) from the internal substances of the living body is detected, so there is a problem that good image quality cannot be obtained. was there. In addition, visible light is also absorbed by substances that make up the body, such as hemoglobin, so the light does not reach deep inside the body, making it difficult to observe blood vessels and organs in living organisms. There was a problem.

By the way, the wavelength region of 700 to 1800 nm is called the biological window and is said to be a region with high biological permeability. At present, indocyanine green, which emits fluorescence at a wavelength of 650 to 900 nm, which is said to be the first optical window of living organisms, is used as a fluorescent labeling agent for fluorescent bioimaging. However, the fluorescent labeling agent that emits fluorescence in the first optical window cannot ignore the adverse effects of autofluorescence and light scattering, and it has been difficult to obtain clear imaging of deep areas.

On the other hand, fluorescence imaging in the region of 1000 to 1400 nm, which is called the second optical window of the living body, is less susceptible to the adverse effects of autofluorescence and scattering from living tissue than the first optical window of the living body. is expected to become possible. Therefore, there is a need for fluorescent labeling agents that exhibit fluorescence in the second optical window of the body in order to achieve sharp and deep imaging. However, since the fluorescent labeling agent using the cyanine dye disclosed in Patent Document 1 has low fluorescence intensity and light resistance, it is difficult to obtain clear imaging, and the fluorescent labeling agent is easily deteriorated by light irradiation. was there.

On the other hand, phthalocyanine dyes are known as dyes having excellent optical properties, high fluorescence intensity, and high light resistance. For example, Patent Document 2 discloses a fluorescent labeling agent containing a phthalocyanine dye. However, all of them have a fluorescence wavelength of less than 1000 nm, and there is a problem that highly sensitive and stable fluorescence bioimaging cannot be obtained. Therefore, in fluorescence imaging, a fluorescent labeling agent that exhibits fluorescence in the second optical window of the living body has been desired in order to perform clear, deep, and stable imaging.

Japanese Unexamined Patent Application Publication No. 2014-231488 JP 2016-65205 A

The problem to be solved by the present invention is to provide a phthalocyanine that emits strong fluorescence in a long-wavelength region of 1000 nm or longer and a fluorescent labeling agent containing the same for performing clear and stable imaging of a deep region in vivo. That is.

As a result of intensive studies, the present inventors have found that the above problems can be solved by extending the conjugation of phthalocyanine and introducing a phosphorus element as a central element, and have completed the present invention.

That is, the present invention relates to a fluorescent labeling agent comprising a phosphorous phthalocyanine represented by the following general formula (1) or general formula (2).

General formula (1)

(Wherein, X ₁ to X ₈ each independently represent a Group 16 element; R ₁ to R ₈ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic Represents an aromatic hydrocarbon group R ₉ to R ₁₂ each independently represents a substituted or unsubstituted aromatic hydrocarbon ring or a substituted or unsubstituted heterocyclic ring R ₁₃ and R ₁₄ each independently hydrogen atom, substituted or unsubstituted aliphatic hydrocarbon group, -P(=O)R ₁₅ R ₁₆ , -C(=O)R ₁₇ , -S(=O) ₂ R ₁₈ , -SiR ₁₉ R ₂₀ R _21. R ₁₅ and R ₁₆ each independently represent a hydrogen atom, a hydroxyl group, a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic hydrocarbon group, a substituted or unsubstituted alkoxyl group, represents a substituted or unsubstituted aryloxy group, R ₁₇ represents a hydrogen atom, a substituted or unsubstituted aliphatic hydrocarbon group, or a substituted or unsubstituted aromatic hydrocarbon group, R ₁₈ represents a hydrogen atom or a hydroxyl group , represents a substituted or unsubstituted aliphatic hydrocarbon group, or a substituted or unsubstituted aromatic hydrocarbon group, and each of R ₁₉ to R ₂₁ independently represents a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted represents a substituted aromatic hydrocarbon group, and Y ₁ ^- represents an anion.)

general formula (2)

(In the formula, X ₉ to X ₁₆ each independently represent a Group 16 element; R ₂₂ to R ₂₉ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic each of R ₃₀ to R ₃₃ independently represents a substituted or unsubstituted aromatic hydrocarbon ring or a substituted or unsubstituted heterocyclic ring R ₃₄ represents a hydrogen atom or a substituted or unsubstituted -P(=O)R ₃₅ R ₃₆ , -C(=O)R ₃₇ , -S(=O) ₂ R ₃₈ , -SiR ₃₉ R ₄₀ R _41. R ₃₅ and Each R ₃₆ independently represents a hydroxyl group, a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic hydrocarbon group, a substituted or unsubstituted alkoxyl group, or a substituted or unsubstituted aryloxy group. R ₃₇ represents a hydrogen atom, a substituted or unsubstituted aliphatic hydrocarbon group or a substituted or unsubstituted aromatic hydrocarbon group, R ₃₈ represents a hydrogen atom, a hydroxyl group or a substituted or unsubstituted aliphatic hydrocarbon R ₃₉ to R ₄₁ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group or a substituted or unsubstituted aromatic hydrocarbon group. )

The present invention also relates to phosphorus phthalocyanine represented by the following general formula (1) or general formula (2).

General formula (1)

(Wherein, X ₁ to X ₈ each independently represent a Group 16 element; R ₁ to R ₈ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic Represents an aromatic hydrocarbon group R ₉ to R ₁₂ each independently represents a substituted or unsubstituted aromatic hydrocarbon ring or a substituted or unsubstituted heterocyclic ring R ₁₃ and R ₁₄ each independently hydrogen atom, substituted or unsubstituted aliphatic hydrocarbon group, -P(=O)R ₁₅ R ₁₆ , -C(=O)R ₁₇ , -S(=O) ₂ R ₁₈ , -SiR ₁₉ R ₂₀ R _21. R ₁₅ and R ₁₆ each independently represent a hydrogen atom, a hydroxyl group, a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic hydrocarbon group, a substituted or unsubstituted alkoxyl group, represents a substituted or unsubstituted aryloxy group, R ₁₇ represents a hydrogen atom, a substituted or unsubstituted aliphatic hydrocarbon group, or a substituted or unsubstituted aromatic hydrocarbon group, R ₁₈ represents a hydrogen atom or a hydroxyl group , represents a substituted or unsubstituted aliphatic hydrocarbon group, or a substituted or unsubstituted aromatic hydrocarbon group, and each of R ₁₉ to R ₂₁ independently represents a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted represents a substituted aromatic hydrocarbon group, and Y ₁ ^- represents an anion.)

general formula (2)

(In the formula, X ₉ to X ₁₆ each independently represent a Group 16 element; R ₂₂ to R ₂₉ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic each of R ₃₀ to R ₃₃ independently represents a substituted or unsubstituted aromatic hydrocarbon ring or a substituted or unsubstituted heterocyclic ring R ₃₄ represents a hydrogen atom or a substituted or unsubstituted -P(=O)R ₃₅ R ₃₆ , -C(=O)R ₃₇ , -S(=O) ₂ R ₃₈ , -SiR ₃₉ R ₄₀ R _41. R ₃₅ and Each R ₃₆ independently represents a hydroxyl group, a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic hydrocarbon group, a substituted or unsubstituted alkoxyl group, or a substituted or unsubstituted aryloxy group. R ₃₇ represents a hydrogen atom, a substituted or unsubstituted aliphatic hydrocarbon group or a substituted or unsubstituted aromatic hydrocarbon group, R ₃₈ represents a hydrogen atom, a hydroxyl group or a substituted or unsubstituted aliphatic hydrocarbon R ₃₉ to R ₄₁ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group or a substituted or unsubstituted aromatic hydrocarbon group. )

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a fluorescent labeling agent capable of performing clear and stable imaging in vivo.

The present invention will be described in detail below, but the present invention is not limited to these.
First, groups in general formulas (1) and (2) are described. First, unsubstituted aliphatic hydrocarbon groups and unsubstituted aromatic hydrocarbon groups for R ₁ to R ₈ and R ₂₂ to R ₂₉ are explained. Both refer to monovalent aliphatic and aromatic hydrocarbon groups.

An alkyl group, a cycloalkyl group, etc. are mentioned as an aliphatic hydrocarbon group.
Here, the alkyl group includes a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, a pentyl group, an isopentyl group, a hexyl group, a heptyl group and an octyl group. , decyl group, dodecyl group, pentadecyl group, octadecyl group and the like. The number of carbon atoms in the alkyl group is preferably 1-18.

Moreover, the cycloalkyl group includes a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, a cyclooctyl group, a cyclooctadecyl group and the like. The cycloalkyl group preferably has 3 to 18 carbon atoms.

Aromatic hydrocarbon groups include aromatic hydrocarbon groups consisting of single rings, condensed rings, ring assemblies and combinations thereof.

Here, the monocyclic aromatic hydrocarbon group includes phenyl group, o-tolyl group, m-tolyl group, p-tolyl group, 2,4-xylyl group, p-cumenyl group, mesityl group and the like. The monocyclic aromatic hydrocarbon group preferably has 6 to 18 carbon atoms.

Further, as the condensed ring aromatic hydrocarbon group, 1-naphthyl group, 2-naphthyl group, 1-anthryl group, 2-anthryl group, 9-anthryl group, 1-phenanthryl group, 9-phenanthryl group, 1-acenaphthyl and the like. The condensed ring aromatic hydrocarbon group preferably has 10 to 18 carbon atoms.

Examples of the ring-assembled aromatic hydrocarbon group include o-biphenylyl group, m-biphenylyl group, p-biphenylyl group and the like. The number of carbon atoms in the ring-assembled aromatic hydrocarbon group is preferably 12-18.

Next, the unsubstituted aromatic hydrocarbon ring and unsubstituted heterocyclic ring for R ₉ to R ₁₂ and R ₃₀ to R ₃₃ will be explained. Each of these means a ring condensed with the phthalocyanine ring represented by general formula (1) or general formula (2). Here, the aromatic hydrocarbon ring includes aromatic hydrocarbon rings such as benzene ring and naphthalene ring.

Moreover, an aliphatic heterocyclic ring and an aromatic heterocyclic ring are mentioned as a heterocyclic ring. Here, the aliphatic heterocycles include aliphatic heterocycles such as imidazolidine ring, 2-imidazoline ring, pyrrolidine ring, 2-pyrroline ring, piperidine ring and piperazine ring.

Moreover, aromatic heterocycles, such as an imidazole ring, a thiazole ring, a pyridine ring, a pyrazine ring, are mentioned as an aromatic heterocycle.
Next, groups for R ₁₃ to R ₂₁ and R ₃₄ to R ₄₁ will be explained. The unsubstituted aliphatic hydrocarbon _group and the unsubstituted aromatic hydrocarbon group _{for R 13 to R 21 and R 34 to R 41 are the unsubstituted} is synonymous with an aliphatic hydrocarbon group and an unsubstituted aromatic hydrocarbon group.

Alkoxyl groups include linear or branched alkoxyl groups, and specific examples include methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, and neopentyloxy groups. , 2,3-dimethyl-3-pentyloxy group, n-hexyloxy group, n-octyloxy group, stearyloxy group, 2-ethylhexyloxy group and the like. The alkoxyl group preferably has 1 to 6 carbon atoms.

Examples of the aryloxy group include aryloxy groups in which the aryl group in the aryloxy group is monocyclic or condensed. A monocyclic aryloxy group is preferred.

Here, the monocyclic aryloxy group includes a phenoxy group, p-methylphenoxy group and the like. Moreover, a naphthyloxy group, an anthryloxy group, etc. are mentioned as an aryloxy group of a condensed ring. The aryloxy group preferably has 6 to 12 carbon atoms.

X ₁ to X ₈ are group 16 elements such as oxygen, sulfur, selenium and tellurium. Among these, oxygen, sulfur and selenium are preferred, and oxygen and sulfur are more preferred in terms of ease of synthesis and stability.
Y ₁ ^— is a monovalent anion, and includes, but is not limited to, halogen ions, hydroxide ions, non-nucleophilic anions, and the like. Examples of non-nucleophilic anions include BF ₄ ⁻ , ^PF ₆ ⁻ , B ₍ C ₆ H ₅ ) ₄ ⁻ , B(C ₆ F ₅ ) ₄ ⁻ and the like. preferable.

The aliphatic hydrocarbon group, aromatic hydrocarbon group, aromatic hydrocarbon ring, aliphatic heterocyclic ring, aromatic heterocyclic ring, alkoxyl group, and aryloxy group described above are further substituted with other substituents. can be Examples of such substituents include halogen atoms, silyl groups, cyano groups, amino groups, and the aforementioned aliphatic hydrocarbon groups, aromatic hydrocarbon groups, alkoxyl groups, and aryloxy groups. In addition, part of the structure may be substituted with an atomic group such as an ether group (ether bond) or a carbonyloxy group (ester bond). For example, substituted aliphatic heterocycles include the following.

Moreover, the following can be mentioned as a substituted aromatic heterocycle.

The phosphorus phthalocyanine of the present invention is characterized by having phosphorus as a central site element and having a structure in which a Group 16 element is bonded to the α-position of the phthalocyanine ring. When an electron-donating group is introduced at the α-position, the orbital energy level of the highest occupied molecular orbital (HOMO) of phthalocyanine rises compared to having no electron-donating group at the α-position, and the lowest unoccupied orbital of phthalocyanine Since the difference (energy gap) between the (LUMO) energy levels becomes smaller, it is considered that absorption and fluorescence wavelengths are exhibited in the long wavelength region of 1000 nm or longer. As the electron-donating group, it is effective to have a structure in which a Group 16 element is directly bonded to the α-position. As such a group 16 element, oxygen, sulfur and selenium are preferable, and oxygen and sulfur are more preferable in terms of ease of synthesis and stability.

The phosphorus phthalocyanine represented by the general formula (1) is obtained by reacting a metal-free phthalocyanine represented by the general formula (3) synthesized by a known method with a phosphorus oxyhalide and then reacting it with an alcohol or an acidic compound. Furthermore, it can be synthesized by reacting with an alkali metal salt.

When the starting phthalonitrile derivative has an asymmetric structure, the obtained phthalocyanine is obtained as a mixture of isomers having different substituent positions. An example of the structure of the isomer is shown herein.

General formula (3)

(Wherein, X ₁ to X ₈ each independently represent a Group 16 element; R ₁ to R ₈ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic represents an aromatic hydrocarbon group, and each of R ₉ to R ₁₂ independently represents a substituted or unsubstituted aromatic hydrocarbon ring or a substituted or unsubstituted heterocyclic ring.)

Phosphorus oxyhalides include phosphorus oxychloride and phosphorus oxybromide. The phosphorus oxyhalide is used in an amount of 1 to 50-fold mol, preferably 5 to 30-fold mol, per 1 mol of the metal-free phthalocyanine represented by the general formula (3). The reaction temperature is 0 to 120°C, preferably 10 to 100°C. The reaction time is 10 minutes to 20 hours, preferably 1 hour to 5 hours. It is preferred to use a solvent in the reaction. Solvents used in the reaction include pyridine, quinoline, triethylamine, dimethylacetamide (DMAC), dimethylformamide (DMF), dimethylimidazolidinone (DMI), dimethylsulfoxide (DMSO) and the like. The amount of the solvent to be used is 1 to 200 times the volume of the metal-free phthalocyanine represented by general formula (3), preferably 10 to 100 times the volume.

The alcohol or acidic compound is used in an amount of 5 to 5,000 mols, preferably 10 to 3,000 mols per 1 mol of the metal-free phthalocyanine represented by the general formula (3). The reaction temperature is 0-65°C, preferably 10-40°C. The reaction time is 5 minutes to 10 hours, preferably 15 minutes to 2 hours. It is preferred to use a solvent in the reaction. Solvents used in the reaction include halogen solvents such as chloroform, dichloromethane, dichloroethane, trichloroethane, chlorobenzene and dichlorobenzene, and aromatic solvents such as benzene, toluene and xylene. The amount of the solvent used is 1 to 300 times the volume of the metal-free phthalocyanine represented by general formula (3), preferably 5 to 200 times the volume. The alkali metal salt is used in an amount of 1 to 10 mol, preferably 1 to 5 mol, per 1 mol of the metal-free phthalocyanine represented by the general formula (3). The reaction temperature is 0-100°C, preferably 10-40°C. The reaction time is 5 minutes to 24 hours, preferably 1 hour to 15 hours.

The phosphorus phthalocyanine represented by the general formula (2) is synthesized by reacting a metal-free phthalocyanine represented by the general formula (4) synthesized by a known method with phosphorus oxyhalide and then reacting it with water. can. Subsequent reaction with alcohols or acidic compounds can introduce axial substituents.

When the starting phthalonitrile derivative has an asymmetric structure, the obtained phthalocyanine is obtained as a mixture of isomers having different substituent positions. An example of the structure of the isomer is shown herein.

general formula (4)

(In the formula, X ₉ to X ₁₆ each independently represent a Group 16 element; R ₂₂ to R ₂₉ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic each of R ₃₀ to R ₃₃ independently represents a substituted or unsubstituted aromatic hydrocarbon ring or a substituted or unsubstituted heterocyclic ring.)

Phosphorus oxyhalides include phosphorus oxychloride and phosphorus oxybromide. The phosphorus oxyhalide is used in an amount of 1 to 50-fold mol, preferably 5 to 30-fold mol, per 1 mol of the metal-free phthalocyanine represented by general formula (4). The reaction temperature is 0 to 120°C, preferably 10 to 100°C. The reaction time is 10 minutes to 48 hours, preferably 5 hours to 30 hours. It is preferred to use a solvent in the reaction. Solvents used in the reaction include pyridine, quinoline, triethylamine, dimethylacetamide (DMAC), dimethylformamide (DMF), dimethylimidazolidinone (DMI), dimethylsulfoxide (DMSO) and the like. The amount of the solvent to be used is 1 to 300 times the volume of the metal-free phthalocyanine represented by general formula (4), preferably 10 to 150 times the volume.

The amount of water to be used is 5 to 5,000 mols, preferably 10 to 3,000 mols per 1 mol of the metal-free phthalocyanine represented by the general formula (4). The alcohol or acidic compound is used in an amount of 5 to 5,000 mols, preferably 10 to 3,000 mols per 1 mol of the metal-free phthalocyanine represented by the general formula (4). The reaction temperature is 0-65°C, preferably 10-40°C. The reaction time is 5 minutes to 10 hours, preferably 15 minutes to 2 hours. It is preferred to use a solvent in the reaction. Solvents used in the reaction include halogen solvents such as chloroform, dichloromethane, dichloroethane, trichloroethane, chlorobenzene and dichlorobenzene, and aromatic solvents such as benzene, toluene and xylene. The amount of the solvent used is 1 to 300 times the volume of the metal-free phthalocyanine represented by general formula (4), preferably 5 to 200 times the volume.

(Fluorescent labeling agent)
The fluorescent labeling agent of the present invention is characterized by comprising a phosphorous phthalocyanine represented by general formula (1) or general formula (2). The use of the fluorescent labeling agent of the present invention is not particularly limited, but examples include visualization of in vivo behavior by fluorescently labeling proteins and low-molecular-weight compounds in vivo. For these uses, the fluorescent labeling agent needs to emit fluorescence in water, and it is preferable that the fluorescent labeling agent is water-soluble or dispersed in water. For example, phosphorus phthalocyanine can be made water-soluble by introducing a sodium sulfonate group or a polyethylene glycol group. It is preferable to use as a dispersion in which the particles containing the particles are dispersed in water.

Furthermore, in order to impart target selectivity, it is preferable to modify the particles containing phosphorus phthalocyanine with a substance that binds to the target molecule. Substances that bind to target molecules include primary antibodies that specifically bind to target molecules, secondary antibodies that bind to the primary antibodies, avidin, biotin, sugar chains, and the like.

(Amphiphile)
When a water-insoluble organic dye is used in an aqueous system, a method of solubilizing it with an amphipathic substance or the like is known. An amphipathic substance is a general term for molecules having a hydrophilic group and a hydrophobic group in one molecule, and typical examples thereof include surfactants and phospholipids. Only one type of amphiphilic substance may be used, or two or more types may be mixed and used. The amphiphilic substance related to this embodiment is not particularly limited, and any amphiphilic substance can be used as long as it can solubilize the phosphorous phthalocyanine of the present invention.

Examples of surfactants include nonionic surfactants, cationic surfactants, anionic surfactants, and the like.
Examples of nonionic surfactants include polyoxyethylene sorbitan fatty acid esters such as Tween (registered trademark) 20, Tween (registered trademark) 40, Tween (registered trademark) 60 and Tween (registered trademark) 80, Cremophor ( EL, polyoxyethylene castor oil derivatives such as Cremophor® RH60, 12-hydroxystearic acid-polyethylene glycol copolymers such as Solutol® HS15, Triton® X-100, Triton® (trademark) X-114 and other octylphenol ethoxylates.

Examples of cationic surfactants include alkyltrimethylammonium salts such as stearyltrimethylammonium chloride and lauryltrimethylammonium chloride, alkylpyridinium salts such as cetylpyridinium chloride, distearyldimethylammonium chloride, dialkyldimethylammonium salts, poly(N ,N'-dimethyl-3,5-methylenepiperidinium), alkyldimethylbenzylammonium salts, alkylisoquinolinium salts, dialkylmorphonium salts, polyoxyethylenealkylamines, alkylamines Salt, polyamine fatty acid derivative, amyl alcohol fatty acid derivative, benzalkonium chloride, benzethonium chloride, polyvinyl alcohol, polyoxyethylene polyoxypropylene glycol, polyethylene glycol-polyalkyl, polyethylene glycol-polylactic acid, polyethylene glycol-polycaprolactone, polyethylene glycol block copolymers such as polyglycolic acid, polyethylene glycol-poly(lactide-glycolide), and the like.

Examples of anionic surfactants include sodium dodecylsulfate, dodecylbenzenesulfonate, decylbenzenesulfonate, undecylbenzenesulfonate, tridecylbenzenesulfonate, nonylbenzenesulfonate and sodium, potassium and ammonium salts thereof.

As long as the fluorescent labeling agent of the present invention contains the phosphorous phthalocyanine of the present invention, it may contain other components as necessary. Other components include, for example, water and amphiphilic substances.

(Method for producing fluorescent labeling agent)
The method for producing the fluorescent labeling agent of the present invention is not particularly limited, but for example, a method of dissolving the phosphorous phthalocyanine of the present invention and an amphiphilic substance in an organic solvent, distilling off the organic solvent, and redissolving in water. Alternatively, a method of injecting water into a solution in which the phosphorus phthalocyanine of the present invention and an amphiphilic substance are dissolved in an organic solvent and distilling off the organic solvent can be used.

The former method is advantageous in that the organic solvent can be easily distilled off and the concentrations of the phosphorus phthalocyanine of the present invention and the amphiphilic substance can be estimated relatively easily. A vacuum distillation device such as an evaporator is preferably used for distilling off the organic solvent. The temperature at which the solvent is distilled off can be arbitrarily set within the range from 15° C. to the boiling point temperature of the organic solvent. When re-dissolving in water, stirring by a propeller stirrer, turbine stirrer, vortex mixer, or magnetic stirrer using a stirrer, or dispersion by an ultrasonic irradiation device, or the like is preferably used. Also, a colloid mill or the like may be used in combination.

The latter method has the advantage of facilitating uniform preparation of micellar particles. When injecting water into a solution dissolved in an organic solvent, it is preferred that the solution is stirred or irradiated with ultrasonic waves, and the water is injected in a short period of time. Stirring can be performed with the same equipment as described above. The temperature at which water is injected can be arbitrarily set within a range from 15° C. to a temperature 5° C. lower than the boiling point of the organic solvent. Evaporation of the organic solvent is preferably carried out under normal pressure while being stirred or irradiated with ultrasonic waves, or under reduced pressure using an evaporator or the like. The temperature at which the solvent is distilled off can be arbitrarily set within the range from 15° C. to the boiling point temperature of the organic solvent.

Examples of organic solvents used in the above production method include hydrocarbons such as hexane, cyclohexane, and heptane, ketones such as acetone and methyl ethyl ketone, ethers such as diethyl ether and tetrahydrofuran, dichloromethane, chloroform, and tetrachloride. Carbon, halogenated hydrocarbons such as dichloroethane and trichloroethane, aromatic hydrocarbons such as benzene and toluene, esters such as ethyl acetate and butyl acetate, N,N-dimethylformamide, dimethylsulfoxide, N-methyl-2-pyrrolidone and aprotic polar solvents such as pyridine derivatives. These solvents may be used alone or in combination. Alternatively, the organic solvent may be removed by dialysis of the fluorescent labeling agent produced by the above method.

As water used in the above preparation method, ion-exchanged water, distilled water, or ultrapure water can be mentioned. When dealing with cells, physiological saline in which salt is added to water to approximate physiological conditions, phosphate-buffered saline in which a phosphate buffer is further added, and the like can be preferably used.

EXAMPLES The present invention will be described below based on examples, but the present invention is not limited by these. In the examples, unless otherwise specified, "part" represents "mole part" and "%" represents "% by mass".

(Production Example 1) Production of phthalonitrile derivative 1 Under a nitrogen atmosphere, 1 part of 1,4-dihydroxy-2,3-naphthalene dicarbonitrile and 2.1 parts of sodium hydride (paraffin-containing purity 60%) were combined with dehydrated dimethylacetamide. Stirred for 15 minutes at 25° C. in 50 parts. After that, 2 parts of ethyl iodide was added, and the mixture was stirred at 80° C. for 9 hours. After cooling, 350 parts of methanol was added, and the precipitate was taken out by filtration and dried at 80° C. to obtain 0.9 parts of phthalonitrile derivative 1.

(Production Examples 2 to 23) Production of phthalonitrile derivatives 2 to 23 1,4-dihydroxy-2,3-naphthalenedicarbonitrile and ethyl iodide used in Production Example 1 are listed in A, shown in Table 1, respectively. Phthalonitrile derivatives 2 to 23 shown in Table 1 were obtained in the same manner as in Production Example 1, except that the compound was changed to the compound listed in B and the compound listed in B. The compounds listed in A and B shown in Table 1 were used in the same molar amounts as 1,4-dihydroxy-2,3-naphthalenedicarbonitrile and ethyl iodide in Production Example 1.

(Production Example 24) Production of phthalonitrile derivative 24 1 part of 1,4-dihydroxy-2,3-naphthalenedicarbonitrile, 4 parts of potassium carbonate and 2 parts of tosyl chloride were mixed in 20 parts of acetone at an internal temperature of 55°C. Reflux for 5 hours. After cooling, 330 parts of water was added, and the mixture was stirred for 1 hour. The precipitate was filtered out and dried at 60°C to obtain 1,4-bis(p-toluenesulfonyloxy)-2,3-naphthalenedicarbonitrile. 0.7 parts of 0.7 parts of 1,4-bis(p-toluenesulfonyloxy)-2,3-naphthalenedicarbonitrile, 1.4 parts of benzenethiol, and 2.8 parts of potassium carbonate in 100 parts of dimethyl sulfoxide at 25° C. 23 Stirred for hours. The reaction was stopped with 330 parts of water and extracted with 80 parts of chloroform. After removing water from the extract with anhydrous magnesium sulfate, column chromatography was performed with ethyl acetate/hexane to obtain 0.3 part of phthalonitrile derivative 24 .

(Production Examples 25 to 39) Production of phthalonitrile derivatives 25 to 39 The 1,4-dihydroxy-2,3-naphthalenedicarbonitrile and benzenethiol used in Production Example 24 are shown in Table 2 and listed in C, respectively. Phthalonitrile derivatives 25 to 39 shown in Table 2 were obtained in the same manner as in Production Example 24 except that the compound and the compound listed in D were changed. The compounds listed in C and D shown in Table 2 were used in the same molar amounts as 1,4-dihydroxy-2,3-naphthalenedicarbonitrile and benzenethiol in Production Example 24.

(Production Example 40) Production of phthalonitrile derivative 40 1 part of 2,6-dimethoxy-4,5-diaminophthalonitrile and 1 part of paraformaldehyde were dissolved in 20 parts of acetonitrile, followed by 30 parts of 30% hydrogen peroxide solution and concentrated hydrochloric acid. 10 parts were added. After stirring at 25° C. for 2 hours, an aqueous solution prepared by dissolving 5 parts of sodium hydrogencarbonate in 500 parts of water was added, and the precipitated solid was collected by filtration and washed with 170 parts of water. After drying at 80°C, 4 parts of dehydrated dimethylacetamide and 0.9 parts of sodium hydride (paraffin-containing purity 60%) were added under nitrogen atmosphere, and stirred at 25°C for 15 minutes. After that, 0.9 part of ethyl iodide was added, and the mixture was stirred at 80° C. for 9 hours. After cooling, 170 parts of water was added, and the precipitate was taken out by filtration and dried at 80° C. to obtain 0.8 parts of phthalonitrile derivative 40 .

(Production Examples 41 to 43) Production of phthalonitrile derivatives 41 to 43 2,6-dimethoxy-4,5-diaminophthalonitrile, paraformaldehyde, and ethyl iodide used in Production Example 40 are shown in Table 3, E Phthalonitrile derivatives 41 to 43 shown in Table 3 were obtained in the same manner as in Production Example 40, except that the compounds listed in 1, the compounds listed in F, and the compounds listed in G were changed. In addition, the compounds listed in E, the compounds listed in F, and the compounds listed in G shown in Table 3 are 2,6-dimethoxy-4,5-diaminophthalonitrile, paraformaldehyde, and ethyl iodide in Production Example 40. was used in the same molar amount as

(Production Example 44) Production of phthalonitrile derivative 44 1 part of 2,6-dimethoxy-4,5-diaminophthalonitrile and 1 part of 1,1'-carbonyldiimidazole were dissolved in 20 parts of acetonitrile and mixed with 30% hydrogen peroxide. 30 parts of water and 10 parts of concentrated hydrochloric acid were added. After stirring at 25° C. for 2 hours, an aqueous solution prepared by dissolving 5 parts of sodium hydrogencarbonate in 500 parts of water was added, and the precipitated solid was collected by filtration and washed with 170 parts of water. After drying at 80°C, 4 parts of dehydrated dimethylacetamide and 2.0 parts of sodium hydride (paraffin-containing purity 60%) were added under nitrogen atmosphere and stirred at 25°C for 15 minutes. After that, 2.0 parts of ethyl iodide was added, and the mixture was stirred at 80° C. for 9 hours. After cooling, 170 parts of water was added, and the precipitate was taken out by filtration and dried at 80°C to obtain 0.8 parts of phthalonitrile derivative 44.

(Production Examples 45 to 47) Production of phthalonitrile derivatives 45 to 47 2,6-dimethoxy-4,5-diaminophthalonitrile and ethyl iodide used in Production Example 44 are shown in Table 4 and listed as H. Phthalonitrile derivatives 45 to 47 shown in Table 4 were obtained in the same manner as in Production Example 44 except that the compound and the compound listed in I were changed. The compounds listed for H and the compounds listed for I shown in Table 4 were used in the same molar amounts as 2,6-dimethoxy-4,5-diaminophthalonitrile and ethyl iodide in Production Example 44.

(Production Example 48) Production of metal-free phthalocyanine 1 1-To a lithium solution prepared by dissolving 7 parts of metallic lithium in 1 part of butanol, 1 part of phthalonitrile derivative 1 was added, and the mixture was stirred at an internal temperature of 110°C for 5.5 hours. bottom. After adding 10 parts of methanol and 1 part of hydrochloric acid and stirring at room temperature for 3 hours, the precipitate was taken out by filtration and dried at 60° C. to obtain 0.2 parts of metal-free phthalocyanine 1 having the structure shown in Table 5. rice field.

(Production Examples 49 to 94) Production of metal-free phthalocyanines 2 to 47 In the same manner as in Production Example 48, except that the phthalonitrile derivative 1 used in Production Example 48 was changed to an equimolar part of the phthalonitrile derivative shown in Table 5. were prepared to give the corresponding metal-free phthalocyanines 2-47, respectively.

(Example 1) Production of phosphorus phthalocyanine 1 Under a nitrogen atmosphere, 1 part of metal-free phthalocyanine 1 was dissolved in 10 parts of pyridine, 15.7 parts of phosphoric acid oxybromide was added, and the mixture was stirred at 80°C for 3 hours. After cooling, pyridine was removed using an evaporator, 150 parts of ethanol and 50 parts of dichloromethane were added, and the mixture was stirred at 25° C. for 1 hour. After that, 15 parts of potassium hexafluorophosphate was added, and the mixture was stirred at 25° C. for 12 hours. Recrystallization was performed using dichloromethane and hexane to obtain 0.4 parts of phosphorus phthalocyanine 1 having the structure shown in Table 6. The structure of the obtained phosphorous phthalocyanine 1 was identified by mass spectroscopy (Autoflex II, manufactured by Bruker Daltonics). In the mass spectrum, molecular ion peaks were detected at m/z (positive) = 1187.32 (theoretical value 1186.30) and m/z (negative) = 143.54 (theoretical value 114.96). It was confirmed to have the structure of phosphorus phthalocyanine 1 described.

(Examples 2 to 47) Production of phosphorus phthalocyanines 2 to 47 Except that the metal-free phthalocyanine 1 and ethanol used in Example 1 were changed to the metal-free phthalocyanine and the compounds listed in J shown in Table 6, the following procedures were carried out. Phosphorus phthalocyanines 2 to 47 shown in Table 6 were obtained in the same manner as in Example 1. The metal-free phthalocyanine shown in Table 6 and the compound listed in J were used in the same molar amounts as the metal-free phthalocyanine 1 and ethanol in Example 1. The structures of the obtained phosphorus phthalocyanines 2 to 47 were identified by mass spectroscopy and confirmed to have the structures shown in Table 6. Table 7 shows the results of mass spectrum analysis.

(Example 48) Production of phosphorus phthalocyanine 48 Under a nitrogen atmosphere, 1 part of metal-free phthalocyanine 1 was dissolved in 10 parts of pyridine, 15.7 parts of phosphoric acid oxybromide was added, and the mixture was stirred at 80°C for 3 hours. After cooling, pyridine was removed using an evaporator, 10 parts of diphenyl phosphate and 150 parts of dimethylsulfoxide were added, reacted at 85° C. for 3 hours, and then the reaction solution was added to 150 parts of water. The precipitate was filtered and dissolved in 80 parts of dichloromethane. After that, 15 parts of potassium hexafluorophosphate was added, and the mixture was stirred at 25° C. for 12 hours. Recrystallization was performed using dichloromethane and hexane to obtain 0.1 part of phosphorus phthalocyanine 48 having the structure shown in Table 8. The structure of the obtained phosphorus phthalocyanine 48 was identified by mass spectroscopy. In the mass spectrum, molecular ion peaks were detected at m/z (positive) = 1595.57 (theoretical value 1594.54) and m/z (negative) = 143.87 (theoretical value 144.96). It was confirmed to have the structure of phosphorus phthalocyanine 48 described.

(Examples 49 to 73) Production of phosphorus phthalocyanines 49 to 73 The metal-free phthalocyanine 1, diphenyl phosphate, and potassium hexafluorophosphate used in Example 48 were converted into metal-free phthalocyanine, K, and F, respectively, shown in Table 8. Phosphorus phthalocyanines 49 to 73 shown in Table 8 were obtained in the same manner as in Example 48, except that the listed compounds were used. The metal-free phthalocyanine and the compounds listed for K and F shown in Table 8 were used in the same molar amounts as the metal-free phthalocyanine 1, diphenyl phosphate and potassium hexafluorophosphate in Example 48. The structures of the obtained phosphorous phthalocyanines 49-73 were identified by mass spectroscopy and confirmed to have the structures shown in Table 8. Table 9 shows the results of mass spectrum analysis.

(Example 74) Production of phosphorus phthalocyanine 74 Under a nitrogen atmosphere, 1 part of metal-free phthalocyanine 1 was dissolved in 10 parts of pyridine, 15.7 parts of phosphoric acid oxybromide was added, and the mixture was stirred at 80°C for 1 hour. Next, the reactant was cooled to 25° C., 200 parts of water was added, and the precipitated solid was filtered and washed with 200 parts of water. After that, 200 parts of ion-exchanged water was added and stirred for 2 hours. After filtration, it was dried at 80° C. to obtain 0.6 parts of phosphorus phthalocyanine 74 shown in Table 10. The structure of the obtained phosphorus phthalocyanine 74 was identified by mass spectroscopy. In the mass spectrum, a molecular ion peak was detected at m/z (positive)=1129.98 (theoretical value: 1129.18), confirming that the compound had the structure of phosphorus phthalocyanine 74 shown in Table 10.

(Examples 75 to 77) Production of phosphorus phthalocyanines 75 to 77 Except that the metal-free phthalocyanine 1 and ion-exchanged water used in Example 74 were changed to the metal-free phthalocyanine and compounds listed in G shown in Table 10, Phosphorus phthalocyanines 75 to 77 shown in Table 10 were obtained in the same manner as in Example 74. The metal-free phthalocyanine and the compound listed as G shown in Table 10 were used in the same molar amounts as the metal-free phthalocyanine 1 and ion-exchanged water in Example 74. The structures of the obtained phosphorus phthalocyanines 75-77 were identified by mass spectroscopy and confirmed to have the structures shown in Table 10. Table 11 shows the results of mass spectrum analysis.

(Example 78) Production of phosphorus phthalocyanine 78 Under a nitrogen atmosphere, 1 part of metal-free phthalocyanine 1 was dissolved in 10 parts of pyridine, 15.7 parts of phosphoric acid oxybromide was added, and the mixture was stirred at 80°C for 1 hour. Next, the reactant was cooled to 25° C., 200 parts of water was added, and the precipitated solid was filtered and washed with 200 parts of water. After dissolving in 150 parts of dimethyl sulfoxide and adding 10 parts of diphenyl phosphate, reaction was carried out at 90° C. for 6 hours. Thereafter, the reactant was cooled to 25° C., 200 parts of water was added, and the precipitated solid was filtered and washed with 200 parts of water. Drying at 80° C. gave 0.6 parts of phosphorus phthalocyanine 78 shown in Table 10. The structure of the obtained phosphorus phthalocyanine 78 was identified by mass spectroscopy. In the mass spectrum, a molecular ion peak was detected at m/z (positive)=1362.24 (theoretical value: 1361.35), confirming that the compound had the structure of phosphorus phthalocyanine 78 shown in Table 12.

(Examples 79 to 82) Preparation of phosphorus phthalocyanines 79 to 82 Except for changing the metal-free phthalocyanine and diphenyl phosphate used in Example 78 to the metal-free phthalocyanine and the compounds listed in H shown in Table 12, the following steps were carried out. The phosphorus phthalocyanines 79 to 82 shown in Table 12 were obtained by the same preparation as in Example 78. The metal-free phthalocyanine and the compound listed for H shown in Table 12, respectively, were used in the same molar amounts as the metal-free phthalocyanine and diphenyl phosphate in Example 78. The structures of the obtained phosphorous phthalocyanines 79-82 were identified by mass spectroscopy and confirmed to have the structures shown in Table 12. Table 13 shows the results of mass spectrum analysis.

(Example 83) Preparation of fluorescent labeling agent D-1 containing phosphorous phthalocyanine 1 part by mass of phosphorous phthalocyanine 1, polyethylene glycol (PEG)/polycaprolactone (PCL) block copolymer (manufactured by Sigma-Aldrich Co., Ltd.) as an amphipathic substance , PEG molecular weight 5000, PCL molecular weight 5000) was dissolved in 20000 parts by weight of acetone. 25000 parts by mass of water was added dropwise to the obtained acetone solution while stirring at 25°C. Furthermore, after acetone was distilled off by stirring at 25 ° C. for 12 hours, it was filtered through a 0.45 μm nylon membrane filter, and the phosphor phthalocyanine 1 was solubilized in the PEG-PCL micelles. Agent D-1 was prepared.

(Examples 84 to 164) Preparation of phosphorous phthalocyanine-containing fluorescent labeling agents D-2 to D-82 Similarly, dispersions D2 to D82 corresponding to each were obtained. Phosphorus phthalocyanine was used in the same molar amount as phosphorus phthalocyanine 1 in Example 83.

(Comparative Example 1) Preparation of Fluorescent Labeling Material S-1 As a dye that emits fluorescence in a wavelength region of 1000 nm or more, phosphorus phthalocyanine was used as a comparative compound, cyanine dye IR1061 (2,6-diphenyl-4-[2-[2- Chloro-3-[2-(2,6-diphenyl-1-thia-2,5-cyclohexadien-4-ylidene)ethylidene]-1-cyclohexenyl]ethenyl]-1-thiabenzene-1-ium tetra Dispersion S-1 was obtained in the same manner as in Example 83, except that it was changed to fluoroborate). IR1061 was used in the same molar amount as phosphorus phthalocyanine 1 in Example 83.

(Evaluation of fluorescence intensity of fluorescent labeling agent)
HeLa cells were seeded in a 96-well plate (5 × 10 ³ cells/well), and 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin-containing Minimum Essential Media (MEM) was used to incubate (37 °C, 5% _CO2 -containing Air, humidified environment) for 24 hours. Thereafter, the medium was removed, and each dispersion prepared in Examples 83 to 164 and Comparative Example 1 was diluted 10-fold with water, added, left to stand for 24 hours in an incubator, and then phosphate-buffered saline ( PBS). Fluorescence intensity was evaluated using an in vivo fluorescence imaging system (SAI-1000 manufactured by Shimadzu Corporation). When the maximum fluorescence intensity of Dispersion S-1 is assumed to be 1, the fluorescence intensity of each fluorescent labeling agent was calculated. When the relative value of the fluorescence intensity is 2 or more, it can be said that each phosphorous phthalocyanine has good properties as a fluorescent labeling agent. Table 15 shows the fluorescence intensity evaluation results.
△: fluorescence intensity 1 or more, less than 2 (defective)
○: fluorescence intensity 2 or more, less than 4 (good)
◎: fluorescence intensity of 4 or more (extremely good)

(Evaluation of light resistance of fluorescent labeling agent)
For the prepared dispersions D-1 to D-82 and S-1, in order to evaluate the photostability, an in vivo fluorescence imaging system (manufactured by Shimadzu Corporation, SAI-1000) was used to irradiate light for 30 minutes. Absorption spectra before and after irradiation were measured using a spectrophotometer (U-4100, manufactured by Hitachi High-Technologies Corporation). The absorbance _I0 at the maximum absorption wavelength before light irradiation and the absorbance _I1 at the maximum absorption wavelength after light irradiation were recorded, and the _I1 / _I0 value was used as an index of light resistance. If the evaluation is 0, it is good. Table 11 shows the light resistance evaluation results.
○: I ₁ /I ₀ is 0.9 or more (good)
×: I ₁ /I ₀ is less than 0.9 (defective)

Dispersions D-1 to D-82 (Example 83164) prepared using the phosphorus phthalocyanine of the present invention were all observed to emit strong fluorescence in the long wavelength region of 1000 nm or longer. Fluorescence intensity and light resistance were better than those of Comparative Example 1). This demonstrates that the phthalocyanine of the present invention has excellent properties as a fluorescent labeling agent.

Claims (2)

A fluorescent labeling agent comprising a phosphorus phthalocyanine represented by the following general formula (1) or (2).
General formula (1)

(Wherein, X ₁ to X ₈ each independently represent a Group 16 element; R ₁ to R ₈ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic R ₉ to R ₁₂ each independently represent a substituted or unsubstituted aromatic hydrocarbon ring or a substituted or unsubstituted heterocyclic ring.
R ₁₃ and R ₁₄ each independently represent a hydrogen atom, a substituted or unsubstituted aliphatic hydrocarbon group, -P(=O)R ₁₅ R ₁₆ , -C(=O)R ₁₇ , -S(=O ) ₂ R ₁₈ , —SiR ₁₉ R ₂₀ R ₂₁ . R ₁₅ and R ₁₆ each independently represent a hydrogen atom, a hydroxyl group, a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic hydrocarbon group, a substituted or unsubstituted alkoxyl group, a substituted or unsubstituted represents an aryloxy group of R ₁₇ represents a hydrogen atom, a substituted or unsubstituted aliphatic hydrocarbon group, or a substituted or unsubstituted aromatic hydrocarbon group. R ₁₈ represents a hydrogen atom, a hydroxyl group, a substituted or unsubstituted aliphatic hydrocarbon group, or a substituted or unsubstituted aromatic hydrocarbon group. R ₁₉ to R ₂₁ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group or a substituted or unsubstituted aromatic hydrocarbon group. Y ₁ ^- represents an anion. )

general formula (2)

(In the formula, X ₉ to X ₁₆ each independently represent a Group 16 element; R ₂₂ to R ₂₉ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic R ₃₀ to R ₃₃ each independently represent a substituted or unsubstituted aromatic hydrocarbon ring or a substituted or unsubstituted heterocyclic ring.
R ₃₄ is a hydrogen atom, a substituted or unsubstituted aliphatic hydrocarbon group, -P(=O)R ₃₅ R ₃₆ , -C(=O)R ₃₇ , -S(=O) ₂ R ₃₈ , -SiR ₃₉ R ₄₀ R ₄₁ . R ₃₅ and R ₃₆ each independently represent a hydroxyl group, a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic hydrocarbon group, a substituted or unsubstituted alkoxyl group, a substituted or unsubstituted aryloxy represents a group. _R37 represents a hydrogen atom, a substituted or unsubstituted aliphatic hydrocarbon group, or a substituted or unsubstituted aromatic hydrocarbon group. _R38 represents a hydrogen atom, a hydroxyl group, a substituted or unsubstituted aliphatic hydrocarbon group, or a substituted or unsubstituted aromatic hydrocarbon group. R ₃₉ to R ₄₁ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group or a substituted or unsubstituted aromatic hydrocarbon group. ) Phosphorus phthalocyanine represented by the following general formula (1) or general formula (2).

General formula (1)

(Wherein, X ₁ to X ₈ each independently represent a Group 16 element; R ₁ to R ₈ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic Represents an aromatic hydrocarbon group R ₉ to R ₁₂ each independently represents a substituted or unsubstituted aromatic hydrocarbon ring or a substituted or unsubstituted heterocyclic ring R ₁₃ and R ₁₄ each independently hydrogen atom, substituted or unsubstituted aliphatic hydrocarbon group, -P(=O)R ₁₅ R ₁₆ , -C(=O)R ₁₇ , -S(=O) ₂ R ₁₈ , -SiR ₁₉ R ₂₀ R _21. R ₁₅ and R ₁₆ each independently represent a hydrogen atom, a hydroxyl group, a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic hydrocarbon group, a substituted or unsubstituted alkoxyl group, represents a substituted or unsubstituted aryloxy group, R ₁₇ represents a hydrogen atom, a substituted or unsubstituted aliphatic hydrocarbon group, or a substituted or unsubstituted aromatic hydrocarbon group, R ₁₈ represents a hydrogen atom or a hydroxyl group , represents a substituted or unsubstituted aliphatic hydrocarbon group, or a substituted or unsubstituted aromatic hydrocarbon group, and each of R ₁₉ to R ₂₁ independently represents a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted represents a substituted aromatic hydrocarbon group, and Y ₁ ^- represents an anion.)

general formula (2)

(In the formula, X ₉ to X ₁₆ each independently represent a Group 16 element; R ₂₂ to R ₂₉ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic each of R ₃₀ to R ₃₃ independently represents a substituted or unsubstituted aromatic hydrocarbon ring or a substituted or unsubstituted heterocyclic ring R ₃₄ represents a hydrogen atom or a substituted or unsubstituted -P(=O)R ₃₅ R ₃₆ , -C(=O)R ₃₇ , -S(=O) ₂ R ₃₈ , -SiR ₃₉ R ₄₀ R _41. R ₃₅ and Each R ₃₆ independently represents a hydroxyl group, a substituted or unsubstituted aliphatic hydrocarbon group, a substituted or unsubstituted aromatic hydrocarbon group, a substituted or unsubstituted alkoxyl group, or a substituted or unsubstituted aryloxy group. R ₃₇ represents a hydrogen atom, a substituted or unsubstituted aliphatic hydrocarbon group or a substituted or unsubstituted aromatic hydrocarbon group, R ₃₈ represents a hydrogen atom, a hydroxyl group or a substituted or unsubstituted aliphatic hydrocarbon R ₃₉ to R ₄₁ each independently represent a substituted or unsubstituted aliphatic hydrocarbon group or a substituted or unsubstituted aromatic hydrocarbon group.