Fluorescent markers and compounds

Abstract

The present application relates to a fluorescent marker and compound. The fluorescent marker comprises a fluorescent pigment represented by the following general formula (1). General formula (1): Q-Z-R1-R2-R3 (wherein, Q represents the residue of the fluorescent pigment. Z represents a direct bond, an alkylene group, or an arylene group. R1 represents a direct bond, ‑O-, ‑OP(=O)R4-, ‑OC(=O)-, ‑OS(=O)2-, ‑OSiR5R6-, ‑C(=O)-, or ‑C(=O)NH-. R2 represents a group selected from the group consisting of an alkylene group, an arylene group, and a heterocyclic group, or a group composed of these groups. R3 represents ‑COOM1, ‑NR7R8, or ‑N + R9R 10 R 11 Here, R4 represents a hydrogen atom, a hydroxyl group, an alkyl group, an aryl group, an alkoxy group, an aryloxy group, or a heterocyclic group. R5 and R6 each independently represent an alkyl group, an aryl group, or a heterocyclic group. 11 (M1 represents a monovalent cation).

Description

Technical Field

[0001] Embodiments of the present invention relate to a fluorescent marking agent and a fluorescent pigment used in the fluorescent marking agent. Background Art

[0002] Bioimaging is a technology that visualizes proteins, cells, and tissues within a living organism. It is widely used in biological and medical research fields, such as elucidating the functions of molecules and cells within a living organism and pharmaceutical research.

[0003] Among them, fluorescence bioimaging is an imaging method that enables dynamic observation of phenomena, multi-color observation, and high-sensitivity observation. In recent years, fluorescence bioimaging has also attracted attention as a non-invasive diagnostic imaging method, with promising applications in clinical practice, such as imaging diagnosis with minimal burden on patients and real-time diagnosis during surgery.

[0004] Fluorescence bioimaging is a method of visualizing a target using a fluorescent dye that specifically binds to a target substance or accumulates at the target site. In this method, the fluorescence emitted by the fluorescent dye when it is irradiated with light in the ultraviolet to near-infrared region is detected.

[0005] Compared to binding-type fluorescence bioimaging, which relies on specific binding to the target substance, the labeling method using aggregation-type fluorescence bioimaging, which involves the accumulation of fluorescence at the target site, is simpler and faster. Furthermore, aggregation-type fluorescence bioimaging offers the advantage of requiring no specific binding to the target substance, resulting in a longer waiting time for fluorescence intensity to stabilize, minimizing the impact on the target substance.

[0006] Patent Documents 1 and 2 disclose an aggregation-type fluorescent dye characterized by being aggregated in phospholipids forming cell membranes.

[0007] Phospholipids form the surface of various biological substances such as cells, liposomes, and extracellular vesicles. In recent years, as used for liposome imaging of drug delivery systems (DDS) and exosome imaging described in non-patent literature 1, the imaging of microscopic substances with phospholipids has attracted attention. In order to perform such in vitro and in vivo imaging, the fluorescent pigments disclosed in patent literature 1 and patent literature 2 are used, and there is a problem of low fluorescence intensity.

[0008] Prior art literature

[0009] Patent Literature

[0010] Patent Document 1: Japanese Patent Application Laid-Open No. 2009-524580

[0011] Patent Document 2: Japanese Patent Application Laid-Open No. 2008-209361

[0012] Non-technical literature

[0013] Non-Patent Document 1: Drug Delivery System, Vol. 29, No. 2, published on March 25, 2014, pp. 116-124 Summary of the Invention

[0014] Problems to be solved by the invention

[0015] In view of the above circumstances, an embodiment of the present invention provides a fluorescent dye that has excellent aggregation properties to phospholipids and exhibits high fluorescence intensity, particularly a fluorescence intensity suitable for a fluorescent labeling agent used for in vitro and in vivo imaging.

[0016] Technical means to solve the problem

[0017] The present inventors have diligently researched and studied to solve the above-mentioned problems, and as a result, have discovered an excellent fluorescent pigment, thereby completing the present invention. That is, embodiments of the present invention relate to the following. However, the present invention is not limited to the following embodiments and includes various embodiments.

[0018] One embodiment relates to a fluorescent labeling agent comprising a fluorescent dye represented by the following general formula (1).

[0019] General formula (1):

[0020] QZ-R1-R2-R3

[0021] In the formula, Q represents the residue of the fluorescent pigment.

[0022] Z represents a direct bond, a substituted or unsubstituted alkylene group, or a substituted or unsubstituted arylene group.

[0023] R1 represents a direct bond, -O-, -OP(=O)R4-, -OC(=O)-, -OS(=O)2-, -OSiR5R6-, -C(=O)-, or -C(=O)NH-.

[0024] R2 represents one group selected from the group consisting of a substituted or unsubstituted alkylene group, a substituted or unsubstituted arylene group, and a substituted or unsubstituted heterocyclic group, or a group consisting of a combination of these groups.

[0025] R3 represents -COOM1, -NR7R8, -N + R9R 10 R11 , -OM2, or -P(=O)(OM3)OM4.

[0026] In the above, R4 represents a hydrogen atom, a hydroxyl group, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted alkoxy group, a substituted or unsubstituted aryloxy group, or a substituted or unsubstituted heterocyclic group.

[0027] R5 and R6 each independently represent a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, or a substituted or unsubstituted heterocyclic group.

[0028] R7~R 11 Each independently represents a hydrogen atom, a substituted or unsubstituted alkyl group, or a substituted or unsubstituted aryl group.

[0029] M1, M2, M3, and M4 each independently represent a hydrogen atom or a monovalent cation.

[0030] In one embodiment, the fluorescent labeling agent is preferably a phospholipid aggregation type fluorescent labeling agent.

[0031] In one embodiment, the fluorescent dye preferably includes a phthalocyanine dye represented by the following general formula (2).

[0032] General formula (2):

[0033]

[0034] Where, X1~X 16 Each independently represents -Z-R1-R2-R3, a hydrogen atom, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted alkenyl group, a substituted or unsubstituted heterocyclic group, -AB, -SO3M5, or -COOM6.

[0035] In the above, A represents a Group 16 element. B represents a hydrogen atom, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted cycloalkyl group, or a substituted or unsubstituted heterocyclic group. M5 and M6 each independently represent a monovalent cation.

[0036] X1~X 16 Adjacent substituents may be linked to each other to form a ring.

[0037] X 17 represents -Z-R1-R2-R3, hydroxyl, halogen, substituted or unsubstituted alkoxy, substituted or unsubstituted aryloxy, -OP(=O)X 18 X 19、-OC(=O)X 20 、-OS(=O)2X 21 , or -OSiX 22 X 23 X 24 .

[0038] In the above, X 18 and X 19 Each independently represents a hydrogen atom, a hydroxyl group, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted alkoxy group, a substituted or unsubstituted aryloxy group, or a substituted or unsubstituted heterocyclic group.

[0039] X 20 represents a hydrogen atom, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, or a substituted or unsubstituted heterocyclic group.

[0040] X 21 represents a hydroxyl group, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, or a substituted or unsubstituted heterocyclic group.

[0041] X 22 ~X 24 Each independently represents a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, or a substituted or unsubstituted heterocyclic group.

[0042] Y represents a divalent to pentavalent metal atom, and k is an integer. When Y is a divalent metal atom, k is 0, when Y is a trivalent metal atom, k is 1, and when Y is a tetravalent or pentavalent metal atom, k is 2.

[0043] Among them, X1~X 17 At least one of them is -Z-R1-R2-R3.

[0044] In one embodiment, the fluorescent marker preferably comprises X of the general formula (2) 17 It is a fluorescent pigment of -Z-R1-R2-R3.

[0045] One embodiment relates to a compound represented by the following general formula (3).

[0046] General formula (3):

[0047]

[0048] Where, X1~X 16Each independently represents -Z-R1-R2-R3, a hydrogen atom, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted alkenyl group, a substituted or unsubstituted heterocyclic group, -AB, -SO3M5, or -COOM6.

[0049] In the above, A represents a Group 16 element. B represents a hydrogen atom, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted cycloalkyl group, or a substituted or unsubstituted heterocyclic group. M5 and M6 each independently represent a monovalent cation.

[0050] X1~X 16 Adjacent substituents may be linked to each other to form a ring.

[0051] X 17 represents -Z-R1-R2-R3, hydroxyl, halogen, substituted or unsubstituted alkoxy, substituted or unsubstituted aryloxy, -OP(=O)X 18 X 19 、-OC(=O)X 20 、-OS(=O)2X 21 , or -OSiX 22 X 23 X 24 .

[0052] In the above, X 18 and X 19 Each independently represents a hydrogen atom, a hydroxyl group, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted alkoxy group, a substituted or unsubstituted aryloxy group, or a substituted or unsubstituted heterocyclic group.

[0053] X 20 represents a hydrogen atom, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, or a substituted or unsubstituted heterocyclic group.

[0054] X 21 represents a hydroxyl group, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, or a substituted or unsubstituted heterocyclic group.

[0055] X 22 ~X 24 Each independently represents a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, or a substituted or unsubstituted heterocyclic group.

[0056] Y represents a divalent to pentavalent metal atom, and k is an integer. When Y is a divalent metal atom, k is 0, when Y is a trivalent metal atom, k is 1, and when Y is a tetravalent or pentavalent metal atom, k is 2.

[0057] Among them, X1~X 17 At least one of them is -Z-R1-R2-R3, as shown below.

[0058] Z represents a direct bond, a substituted or unsubstituted alkylene group, or a substituted or unsubstituted arylene group.

[0059] R1 represents a direct bond, -O-, -OP(=O)R4-, -OC(=O)-, -OS(=O)2-, -OSiR5R6-, -C(=O)-, or -C(=O)NH-.

[0060] R2 represents one group selected from the group consisting of a substituted or unsubstituted alkylene group, a substituted or unsubstituted arylene group, and a substituted or unsubstituted heterocyclic group, or a group consisting of a combination of these groups.

[0061] R3 represents -COOM1, -NR7R8, -N + R9R 10 R 11 , -OM2, or -P(=O)(OM3)OM4.

[0062] In the above, R4 represents a hydrogen atom, a hydroxyl group, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted alkoxy group, a substituted or unsubstituted aryloxy group, or a substituted or unsubstituted heterocyclic group. R5 and R6 each independently represent a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, or a substituted or unsubstituted heterocyclic group. R7 to R 11 Each independently represents a hydrogen atom, a substituted or unsubstituted alkyl group, or a substituted or unsubstituted aryl group. M1, M2, M3, and M4 each independently represent a hydrogen atom or a monovalent cation.

[0063] The disclosure of this application is related to the subject matter described in Japanese Patent Application No. 2020-066924 filed on April 2, 2020, the entire disclosure of which is incorporated herein by reference.

[0064] Effects of the Invention

[0065] According to an embodiment of the present invention, by introducing a functional group that has a high ability to aggregate to phospholipids, it is possible to provide a fluorescent dye having a fluorescence intensity suitable for use as a fluorescent labeling agent for in vitro and in vivo imaging. BRIEF DESCRIPTION OF THE DRAWINGS

[0066] Figure 1 This is a graph showing the evaluation results of the fluorescence intensities of the fluorescent labeling agent 1, the fluorescent labeling agent 15, the fluorescent labeling agent 19, the fluorescent labeling agent 24, the fluorescent labeling agent 25, the fluorescent labeling agent 68, and the fluorescent labeling agent 75.

[0067] Figure 2 This is a fluorescence microscope photograph of cells labeled with fluorescent labeling agent 1.

[0068] Figure 3 This is a fluorescence microscope photograph of cells labeled with the fluorescent labeling agent 15 .

[0069] Figure 4 This is a fluorescence microscope photograph of cells labeled with the fluorescent labeling agent 19.

[0070] Figure 5 This is a fluorescence microscope photograph of cells labeled with the fluorescent labeling agent 24 .

[0071] Figure 6 This is a fluorescence microscope photograph of cells labeled with the fluorescent labeling agent 25.

[0072] Figure 7 This is a fluorescence microscope photograph of cells labeled with the fluorescent labeling agent 68.

[0073] Figure 8 This is a fluorescence microscope photograph of cells labeled with the fluorescent labeling agent 75.

[0074] Figure 9 This is a fluorescence microscope photograph of cells labeled with the fluorescent labeling agent 42.

[0075] Figure 10 This is a fluorescence microscope photograph of cells labeled with the fluorescent labeling agent 53. DETAILED DESCRIPTION

[0076] Hereinafter, embodiments of the present invention will be described in detail. However, the embodiments of the present invention are not limited to the following description and include various embodiments.

[0077] A fluorescent labeling agent according to one embodiment of the present invention includes a compound represented by the following general formula (1): The compound represented by the following general formula (1) is a fluorescent dye.

[0078] General formula (1):

[0079] QZ-R1-R2-R3

[0080] In the formula, Q represents the residue of a fluorescent pigment. In this specification, a fluorescent pigment refers to a pigment that emits fluorescence when irradiated with light in the ultraviolet region to the near-infrared region (for example, light with a wavelength of 560nm to 900nm), and can be a known compound. As fluorescent pigments, there are no particular limitations, but for example, pigments such as fluoresceins, rhodamines, coumarins, cyanines, phthalocyanines, diketopyrrolopyrroles, boron dipyrromethene (BODIPY), xanthenes, pyrenes, merocyanines, perylenes, acridines, stilbenes, pyrromethenes, and umbelliferones can be listed.

[0081] In one embodiment, the compound represented by the general formula (1) (fluorescent dye) may have, for example, the skeleton of the dye exemplified above as the residue Q of the fluorescent dye. That is, the compound represented by the general formula (1) may be a compound having a structure in which at least one substituent (functional group) represented by -Z-R1-R2-R3 is introduced into the skeleton of the dye exemplified above.

[0082] In one embodiment, from the perspective of stability and fluorescence wavelength, the fluorescent pigment is preferably a phthalocyanine. In one embodiment, a compound represented by the following general formula (2) (phthalocyanine pigment) can be preferably used as a fluorescent pigment. 17 It is a prerequisite that at least one substituent is represented by -Z-R1-R2-R3.

[0083] When the fluorescent dye constituting the fluorescent labeling agent comprises a compound represented by the following general formula (2), a fluorescent labeling agent having a phthalocyanine-derived skeleton and excellent durability can be readily obtained. Furthermore, luminescence at wavelengths suitable for in vitro and in vivo bioimaging (e.g., 650 nm to 900 nm) can be readily obtained from a phthalocyanine-derived skeleton.

[0084] General formula (2):

[0085]

[0086] In the fluorescent dye of the embodiment, "-Z-R1-R2-R3" is a substituent having a hydrophilic group, which can enhance the fluorescent dye's ability to aggregate to phospholipids through electrostatic interaction with the hydrophilic group in phospholipids. The specific structure of the substituent is as follows.

[0087] Z represents a direct bond, a substituted or unsubstituted alkylene group, or a substituted or unsubstituted arylene group. In one embodiment, Z is preferably a direct bond.

[0088] R1 represents a direct bond, -O-, -OP(=O)R4-, -OC(=O)-, -OS(=O)2-, -OSiR5R6-, -C(=O)-, or -C(=O)NH-. In one embodiment, R1 is preferably -OP(=O)R4-, -OS(=O)2-, or -OSiR5R6-.

[0089] R2 represents a group selected from the group consisting of a substituted or unsubstituted alkylene group, a substituted or unsubstituted arylene group, and a substituted or unsubstituted heterocyclic group, or a group composed of a combination of these groups. In one embodiment, R2 is preferably a substituted or unsubstituted alkylene group or a substituted or unsubstituted arylene group, more preferably a substituted or unsubstituted alkylene group. In one embodiment, R2 is preferably an alkylene group. The main chain of the alkylene group preferably has 1 to 10 carbon atoms.

[0090] R3 represents -COOM1, -NR7R8, or -N + R9R 10 R 11 In addition, R3 represents -OM2, or -P(=O)(OM3)OM4. In one embodiment, R3 is preferably -COOM1, -NR7R8, -OM2, or -P(=O)(OM3)OM4.

[0091] In the above, R4 represents a hydrogen atom, a hydroxyl group, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted alkoxy group, a substituted or unsubstituted aryloxy group, or a substituted or unsubstituted heterocyclic group. In one embodiment, R4 is preferably a hydrogen atom, a hydroxyl group, a substituted or unsubstituted alkyl group, or an unsubstituted aryl group.

[0092] R5 and R6 each independently represent a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, or a substituted or unsubstituted heterocyclic group. In one embodiment, R5 and R6 each independently represent a substituted or unsubstituted alkyl group or an unsubstituted aryl group. The alkyl group is more preferably a linear or branched alkyl group having 1 to 5 carbon atoms.

[0093] R7~R 11 Each independently represents a hydrogen atom, a substituted or unsubstituted alkyl group, or a substituted or unsubstituted aryl group. 11 Preferably, each independently represents a hydrogen atom or a substituted or unsubstituted alkyl group. More preferably, the alkyl group is a linear or branched alkyl group having 1 to 5 carbon atoms.

[0094] M1, M2, M3, and M4 each independently represent a hydrogen atom or a monovalent cation. Examples of monovalent cations include alkali metals and quaternary amines. Examples of alkali metals include lithium, sodium, potassium, rubidium, and cesium. In one embodiment, M1, M2, M3, and M4 are each preferably a hydrogen atom.

[0095] X1~X 16 Each independently represents a hydrogen atom, or a substituent selected from the group consisting of -Z-R1-R2-R3, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted heterocyclic group, -AB, -SO3M5, and -COOM6.

[0096] In the above, M5 and M6 each independently represent a monovalent cation. Examples of the monovalent cation include alkali metals and quaternary amines. Examples of the alkali metal include lithium, sodium, potassium, rubidium, and cesium.

[0097] In one embodiment, preferably, X1 to X 16 At least one, preferably four or more, of the dye skeleton are the substituents. In one embodiment, the substituents on the dye skeleton are preferably independently substituted or unsubstituted alkyl groups, substituted or unsubstituted aryl groups, or -AB.

[0098] In the -AB, A represents a Group 16 element. Examples of Group 16 elements include oxygen, sulfur, selenium, and tellurium. In one embodiment, A is preferably oxygen, sulfur, or selenium. In terms of ease of synthesis and stability, oxygen or sulfur is more preferred. In terms of fluorescence intensity, oxygen is further preferred. B represents a hydrogen atom, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted cycloalkyl group, or a substituted or unsubstituted heterocyclic group, each as described above. In one embodiment, B is preferably a hydrogen atom, a substituted or unsubstituted alkyl group, or a substituted or unsubstituted aryl group. Therefore, in one embodiment, -AB is preferably -OR, -OAr, -SR, or -SAr. Here, R represents an alkyl group, and Ar represents an aryl group.

[0099] Y represents a divalent to pentavalent metal atom, and k is an integer. When Y is a divalent metal atom, k is 0, when Y is a trivalent metal atom, k is 1, and when Y is a tetravalent or pentavalent metal atom, k is 2. Examples of divalent metal atoms include Mg, Cu, and Zn. Examples of trivalent metal atoms include Al, Ga, and In. Examples of tetravalent metal atoms include Si, Mn, Sn, Cr, and Zr. Examples of pentavalent metal atoms include P. From the perspective of fluorescence intensity, Y is preferably Al, Si, or P, and more preferably Al. From the perspective of light resistance, Y is preferably Al or Si.

[0100] In one embodiment, X1 to X 16 Adjacent substituents may be linked to form a ring. The ring structure may be any of a cycloalkyl, cycloalkenyl, aryl, and heteroaryl group, forming a condensed ring with the aromatic ring in the phthalocyanine skeleton. The ring structure may further have substituents or may be unsubstituted. The number of carbon atoms in the ring structure may be 2 to 30, preferably 4 to 6. The ring is preferably a 5-membered ring or a 6-membered ring.

[0101] In one embodiment, it is preferred that adjacent substituents are linked to each other to form a phenylene group. In such a case, the phenylene group is bonded to the aromatic ring in the phthalocyanine skeleton to form a naphthalene structure. In other embodiments, adjacent substituents may be linked to each other to form a ring containing a nitrogen atom. In such a case, the ring containing a nitrogen atom is bonded to the aromatic ring in the phthalocyanine skeleton to form, for example, an imidazole structure. The ring structure such as the naphthalene structure or the imidazole structure may also have substituents such as alkyl or aryl groups.

[0102] X 17 represents -Z-R1-R2-R3, hydroxyl, halogen, substituted or unsubstituted alkoxy, substituted or unsubstituted aryloxy, -OP(=O)X 18 X 19 、-OC(=O)X 20 、-OS(=O)2X 21 、-OSiX 22 X 23 X 24 In one embodiment, X 17 Preferably, it is -Z-R1-R2-R3 or hydroxyl. Z, R1, R2, and R3 are as described above.

[0103] In the above, X 18 and X 19 Each independently represents a hydrogen atom, a hydroxyl group, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted alkoxy group, a substituted or unsubstituted aryloxy group, or a substituted or unsubstituted heterocyclic group.

[0104] X 20 represents a hydrogen atom, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, or a substituted or unsubstituted heterocyclic group.

[0105] X 21 represents a hydroxyl group, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, or a substituted or unsubstituted heterocyclic group.

[0106] X 22 ~X 24 Each independently represents a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, or a substituted or unsubstituted heterocyclic group.

[0107] Here, the R4 to R 11 , and X1~X 24 The alkyl groups in can be selected independently. The alkyl group may have a substituent or may be unsubstituted.

[0108] Examples of the alkyl group include linear or branched alkyl groups. Specific examples include methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, octadecyl, isopropyl, isobutyl, isopentyl, 2-ethylhexyl, sec-butyl, tert-butyl, sec-pentyl, tert-pentyl, tert-octyl, and neopentyl. The alkyl group preferably has a carbon number of 1 to 30. The carbon number is more preferably in the range of 1 to 20, and even more preferably in the range of 1 to 10.

[0109] Examples of substituents in the alkyl group include halogen atoms such as fluorine, chlorine, and bromine, hydroxyl groups, amino groups, nitro groups, formyl groups, cyano groups, and carboxyl groups, as well as the aforementioned alkyl groups, aryl groups described below, cycloalkyl groups, and heterocyclic groups. Furthermore, when a portion of the structure is substituted with an amide bond (-NHCO-), an ester bond (-COO-), an ether bond (-O-), a urea bond (-NHCONH-), or a carbamate bond (-NHCOO-), the substituted portion is also considered a "substituent."

[0110] Therefore, a substituted alkyl group refers to an alkyl group substituted with the aforementioned substituent. A substituted alkyl group may be an alkyl group substituted with one or more substituents. For example, specific examples of alkyl groups substituted with a halogen atom include trifluoromethyl, 2,2,2-trifluoroethyl, -(CF2)4CF3, -(CF2)5CF3, -(CF2)6CF3, -(CF2)7CF3, -(CF2)8CF3, trichloromethyl, and 2,2-dibromoethyl.

[0111] Specific examples of the alkyl group substituted with an amide bond include: -CH2-CH2-CH2-NHCO-CH2-CH3, -CH2-CH(-CH3)-CH2-NHCO-CH2-CH3, -CH2-CH2-CH2-NHCO-CH2-CH3, -CH2-CH2-CH2-NHCO-CH2-CH3, -CH2-CH2-CH2-CH2-NHCO-CH2-CH(CH2-CH3)-CH2-CH2-CH2-CH3, -(CH2)5-NHCO-(CH2) 11 -CH3, -CH2-CH2-CH2-C(-NHCO-CH2-CH3)3, etc. The number of carbon atoms in the alkyl group substituted with an amide bond is preferably in the range of 2 to 30. The carbon number is more preferably in the range of 2 to 10, and even more preferably in the range of 2 to 5.

[0112] Specific examples of the alkyl group substituted with an ester bond include -CH2-CH2-CH2-COO-CH2-CH3, -CH2-CH(-CH3)-CH2-COO-CH2-CH3, -CH2-CH2-CH2-OCO-CH2-CH3, -CH2-CH2-CH2-CH2-COO-CH2-CH(CH2-CH3)-CH2-CH2-CH2-CH3, and -(CH2)5-COO-(CH2) 11 -CH3, -CH2-CH2-CH2-CH-(COO-CH2-CH3)2, etc. The number of carbon atoms in the alkyl group substituted with an ester bond is preferably in the range of 2 to 30. The carbon number is more preferably in the range of 2 to 10, and even more preferably in the range of 2 to 5.

[0113] Specific examples of the alkyl group substituted with an ether bond include -CH2-O-CH3, -CH2-CH2-O-CH2-CH3, -CH2-CH2-CH2-O-CH2-CH3, -(CH2-CH2-O) n -CH3 (where n is an integer from 1 to 8), -(CH2-CH2-CH2-O) m The alkyl group substituted with an ether bond preferably has a carbon number in the range of 2 to 30. The carbon number is more preferably in the range of 2 to 10, and even more preferably in the range of 2 to 5.

[0114] Specific examples of the alkyl group substituted with a urea bond (-NHCONH-) include -CH2-NHCONH-CH3, -CH2-CH2-NHCONH-CH2-CH3, -CH2-CH2-CH2-NHCONH-CH2-CH3, -(CH2-CH2-NHCONH)n -CH3 (where n is an integer from 1 to 8), -(CH2-CH2-CH2-NHCONH) m -CH3 (where m is an integer from 1 to 5), -CH2-CH(CH3)-NHCONH-CH2-CH3, -CH2-CH-(NHCONHCH3)2, etc., but are not limited to these. The carbon number of the alkyl group substituted with a urea bond is preferably in the range of 2 to 30. The carbon number is more preferably in the range of 2 to 10, and even more preferably in the range of 2 to 5.

[0115] Specific examples of the alkyl group substituted with a carbamate bond include -CH2-CH2-CH2-NHCOO-CH2-CH3, -CH2-CH(-CH3)-CH2-NHCOO-CH2-CH3, -CH2-CH2-CH2-NHCOO-CH2-CH3, -CH2-CH2-CH2-NHCOO-CH2-CH3, -CH2-CH2-CH2-CH2-NHCOO-CH2-CH(CH2-CH3)-CH2-CH2-CH2-CH3, and -(CH2)5-NHCOO-(CH2) 11 -CH3, -CH2-CH2-CH2-CH-(NHCOO-CH2-CH3)2, etc. The number of carbon atoms in the alkyl group substituted with a carbamate bond is preferably in the range of 2 to 30. The carbon number is more preferably in the range of 2 to 10, and even more preferably in the range of 2 to 5.

[0116] Specific examples of the alkyl group substituted with two or more substituents selected from among an amide bond (-NHCO-), an ester bond (-COO-), an ether bond (-O-), a urea bond (-NHCONH-), and a carbamate bond (-NHCOO-) include -CH2-CH2-NHCO-CH2-CH2-O-CH2-CH(CH2-CH3)-CH2-CH2-CH2-CH3, -CH2-CH2-COO-CH2-CH2-O-CH2-CH2-NHCOO-CH2-CH(CH2-CH3)-CH2-CH2-CH2-CH3, and -CH2-CH2-NHCO-CH2(OCO-CH2)-CH2-. The number of carbon atoms in the alkyl group substituted with two or more substituents selected from among an amide bond (-NHCO-), an ester bond (-COO-), an ether bond (-O-), a urea bond (-NHCONH-), and a carbamate bond (-NHCOO-) is preferably within the range of 3 to 30. The carbon number is more preferably in the range of 3 to 10, and even more preferably in the range of 3 to 5.

[0117] The R4~R 11 , and X1~X 24 The aryl groups in can be selected independently. The aryl group may be substituted or unsubstituted.

[0118] Examples of the aryl group include monocyclic or condensed polycyclic aryl groups. Examples include phenyl, 1-naphthyl, 2-naphthyl, p-biphenyl, m-biphenyl, 2-anthryl, 9-anthryl, 2-phenanthrenyl, 3-phenanthrenyl, 9-phenanthrenyl, 2-fluorenyl, 3-fluorenyl, 9-fluorenyl, 1-pyrenyl, 2-pyrenyl, 3-peryl, o-tolyl, m-tolyl, p-tolyl, 4-methylbiphenyl, terphenyl, 4-methyl-1-naphthyl, 4-tert-butyl-1-naphthyl, 4-naphthyl-1-naphthyl, 6-phenyl-2-naphthyl, 10-phenyl-9-anthryl, spirofluorenyl, and 2-benzocyclobutenyl. The number of carbon atoms in the aryl group is preferably within the range of 6 to 18. More preferably, the number of carbon atoms is within the range of 6 to 10.

[0119] The substituent of the substituted aryl group may be the same as the substituent exemplified as the substituent of the alkyl group.

[0120] X1~X 16 The cycloalkyl groups in the cycloalkyl group can be independently selected. The cycloalkyl group may be substituted or unsubstituted. Examples of the cycloalkyl group include cyclopentyl, cyclohexyl, 2,5-dimethylcyclopentyl, and 4-tert-butylcyclohexyl. The cycloalkyl group preferably has a carbon number in the range of 3 to 12. More preferably, the carbon number is in the range of 3 to 6. The substituents of the substituted cycloalkyl group may be the same as those exemplified as the substituents of the alkyl group.

[0121] X1~X 16 The alkenyl groups in the group may be selected independently. The alkenyl group may have a substituent or may be unsubstituted. Examples of the alkenyl group include linear or branched alkenyl groups. An alkenyl group generally refers to a group having one double bond in its structure. In the present specification, an alkenyl group may have multiple double bonds in its structure. Specific examples of alkenyl groups include vinyl, 1-propenyl, allyl, 2-butenyl, 3-butenyl, isopropenyl, isobutenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 1,3-butadienyl, etc. The number of carbon atoms in the alkenyl group is preferably within the range of 2 to 18. The carbon number is more preferably 2 to 10, and even more preferably 2 to 5. The substituents of the substituted alkenyl group may be the same as the substituents exemplified as the substituents in the alkyl group.

[0122] R2, R4~R 11 , and X1~X 24 The heterocyclic group in can be independently selected. The heterocyclic group may be substituted or unsubstituted.

[0123] Examples of the heterocyclic group include aliphatic heterocyclic groups and aromatic heterocyclic groups. Specific examples include pyridyl, pyrazinyl, piperidinyl, pyranyl, morpholinyl, and acridinyl. In addition, groups represented by the following structural formulas may also be mentioned. The number of carbon atoms (the number of carbon atoms constituting the ring) in the heterocyclic group is preferably 4 to 12. The number of ring members is preferably 5 to 13.

[0124]

[0125] The substituents of the substituted heterocyclic group may be the same as those exemplified as the substituents of the alkyl group. Examples of the substituted heterocyclic group include 3-methylpyridyl, N-methylpiperidyl, and N-methylpyrrolyl.

[0126] R4, X 18 , and X 19 The alkoxy groups in can be selected independently. The alkoxy groups may have a substituent or may be unsubstituted.

[0127] Examples of the alkoxy group include linear or branched alkoxy groups. Specific examples include methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, neopentyloxy, 2,3-dimethyl-3-pentyloxy, n-hexyloxy, n-octyloxy, stearyloxy, and 2-ethylhexyloxy. The alkoxy group preferably has 1 to 6 carbon atoms.

[0128] The substituent of the substituted alkoxy group may be the same as the substituent exemplified as the substituent of the alkyl group.

[0129] The substituents of the substituted alkoxy group may be the same as those exemplified as the substituents of the alkyl group. Specific examples of the substituted alkoxy group include trichloromethoxy, trifluoromethoxy, 2,2,2-trifluoroethoxy, 2,2,3,3-tetrafluoropropoxy, 2,2-bis(trifluoromethyl)propoxy, 2-ethoxyethoxy, 2-butoxyethoxy, 2-nitropropoxy, and benzyloxy.

[0130] R4, X 18 , and X 19 The aryloxy groups in can be selected independently. The aryloxy group may have a substituent or may be unsubstituted.

[0131] Examples of the aryloxy group include monocyclic or condensed polycyclic aryloxy groups. Specific examples include phenoxy, p-methylphenoxy, naphthyloxy, and anthryloxy groups. The aryloxy group is preferably a monocyclic aryloxy group. Furthermore, an aryloxy group having 6 to 12 carbon atoms is preferred.

[0132] The substituent of the substituted aryloxy group may be the same as the substituents exemplified as the substituents of the aryl group, and examples of the substituted aryloxy group include p-nitrophenoxy, p-methoxyphenoxy, 2,4-dichlorophenoxy, pentafluorophenoxy, and 2-methyl-4-chlorophenoxy.

[0133] The alkylene groups in Z and R2 can be independently selected. The alkylene group may be substituted or unsubstituted. Examples of the alkylene group include divalent groups obtained by removing one hydrogen atom from the alkyl group. Specific examples of substituted or unsubstituted alkylene groups include -CH2-CH2-, -CH2-CH2-CH2-NHCO-CH2-CH2-, -CH2-CH2-CH2-OCO-CH2-CH2-, and -CH2-CH2-CH2-O-CH2-CH2-.

[0134] The arylene groups in Z and R2 can be independently selected. The arylene group may be substituted or unsubstituted. Examples of the arylene group include divalent groups obtained by removing one hydrogen atom from an aryl group. The arylene group preferably has a carbon number of 6 to 10. In one embodiment, the arylene group may be a phenylene group or a naphthylene group. Specific examples of substituted or unsubstituted arylene groups include groups represented by the following structural formulas.

[0135]

[0136] One embodiment of the present invention relates to a fluorescent labeling agent comprising the aforementioned fluorescent pigment. This fluorescent labeling agent can be used for fluorescent labeling in biological imaging across a wide range of fields, from biochemical research to medical diagnosis. For example, it can be used for fluorescent labeling in genetic diagnosis, immunodiagnosis, medical development, regenerative medicine, environmental testing, biotechnology, and fluorescence inspection.

[0137] The structure (substituent) represented by -Z-R1-R2-R3 in the fluorescent dye of the fluorescent labeling agent of the embodiment described above functions to interact with phospholipids. Therefore, it is preferably used as a phospholipid-aggregating fluorescent labeling agent. Phospholipid-aggregating fluorescent labeling agents are preferably used as fluorescent labeling agents for staining cell membranes, tracking exosomes, and imaging liposomes for drug delivery systems (DDS).

[0138] In the fluorescent labeling agent of the embodiment, the concentration of the fluorescent pigment is not particularly limited. For example, in the case of treating cells, if the effects on cell dysfunction and proliferation inhibition are taken into account, the concentration of the fluorescent pigment is preferably low. In one embodiment, for example, the concentration of the fluorescent pigment relative to 10,000 cells (cells) / well (well) seeded in a 96-well plate is preferably 100 μM or less. The concentration is more preferably 50 μM or less, and further preferably 10 μM or less. According to the fluorescent labeling agent of the embodiment, due to the excellent agglomeration of phospholipids, even low concentrations of fluorescent pigments can be imaged with high fluorescence intensity. Therefore, for example, even at low concentrations of 2 μM or less, more accurate detection can be performed.

[0139] The fluorescent marking agent of the embodiment described above only needs to contain the fluorescent dye of the embodiment described above, and may contain other components as needed. Such other components may be components well known in the art. For example, solvents and amphiphilic substances may be mentioned.

[0140] The solvent may be water or an organic solvent, more preferably water. Taking into account the solubility of the fluorescent pigment, water and an organic solvent may be mixed and used. For example, the organic solvent is preferably ethanol or dimethyl sulfoxide (DMSO).

[0141] An amphiphilic substance is a general term for compounds having a hydrophilic group and a hydrophobic group within a single molecule. Specific examples include surfactants and phospholipids. A single amphiphilic substance may be used, or two or more may be mixed and used. In the fluorescent marker of the embodiment, the amphiphilic substance is not particularly limited and may be any compound as long as it is a water-insoluble fluorescent pigment that can be dissolved in water and emits fluorescence in the near-infrared region. Specific examples of usable amphiphilic substances are listed below, without particular limitation.

[0142] Examples of the surfactant include nonionic surfactants, cationic surfactants, anionic surfactants, and polymer surfactants.

[0143] Examples of the nonionic surfactant 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; polyoxyethylene castor oil derivatives such as Cremophor (registered trademark) EL and Cremophor (registered trademark) RH60; 12-hydroxystearic acid-polyethylene glycol copolymers such as Solutol (registered trademark) HS15; and octylphenol ethoxylates such as Triton (registered trademark) X-100 and Triton (registered trademark) X-114.

[0144] Examples of the cationic surfactant include alkyltrimethylammonium salts such as stearyltrimethylammonium chloride and lauryltrimethylammonium chloride, alkylpyridinium salts such as cetylpyridinium chloride, distearyldimethylammonium chloride, dialkyldimethylammonium salts, and alkylquaternary ammonium salts such as poly(N,N'-dimethyl-3,5-methylenepiperidinium) chloride, alkyldimethylbenzylammonium salts, alkylisoquinolinium salts, dialkylmorpholinium salts, polyoxyethylenealkylamines, alkylamine salts, polyamine fatty acid derivatives, amyl alcohol fatty acid derivatives, benzalkonium chloride, and anilinium chloride.

[0145] Examples of the anionic surfactant include sodium lauryl sulfate, dodecylbenzenesulfonate, decylbenzenesulfonate, undecylbenzenesulfonate, tridecylbenzenesulfonate, and nonylbenzenesulfonate, as well as sodium, potassium, and ammonium salts thereof.

[0146] Examples of the high molecular surfactant include block copolymers such as polyvinyl alcohol, polyoxyethylene polyoxypropylene glycol, polyethylene glycol-polyalkylene glycol, polyethylene glycol-polylactic acid, polyethylene glycol-polycaprolactone, polyethylene glycol-polyglycolic acid, and polyethylene glycol-poly(lactide-glycolide).

[0147] In one embodiment, the fluorescent labeling agent of the embodiment may include one or more of the compounds exemplified above as amphiphilic substances. However, the fluorescent labeling agent of the embodiment has excellent aggregability to target sites such as phospholipids, thus enabling high-sensitivity detection without the need for an amphiphilic substance.

[0148] In the fluorescent marker of the embodiment, the fluorescent pigment preferably comprises a phthalocyanine pigment. The synthesis method of the phthalocyanine pigment is not particularly limited. For example, first, a pigment (phthalocyanine metal complex) with a phthalocyanine skeleton is synthesized using a phthalonitrile derivative as a raw material and a known method. Then, a component with a substituent (-Z-R1-R2-R3) is added to the pigment, and these are heated and stirred in a dimethyl sulfoxide solvent and reacted. By this reaction, the desired fluorescent pigment can be obtained. As the component with the substituent, for example, an acidic compound in the embodiment described later or a compound represented as an axial ligand can be used.

[0149] When the phthalonitrile derivative used as a raw material has an asymmetric structure, phthalocyanine can be obtained as a mixture of isomers with different substituent positions. The following is only an example of a phthalocyanine structure shown in this specification, but isomers with different substituent positions are not excluded.

[0150] Specific examples of the fluorescent dye according to one embodiment of the present invention include the following: However, the fluorescent dye according to the present invention is not limited to these.

[0151] [Table 1-1]

[0152]

[0153] [Table 1-2]

[0154]

[0155] [Table 1-3]

[0156]

[0157] [Table 1-4]

[0158]

[0159] Among the fluorescent dyes exemplified above, fluorescent dyes 1 to 37 each have a phthalocyanine dye skeleton (dye residue), fluorescent dye 38 has a diketopyrrolopyrrole dye skeleton, and fluorescent dyes 39 and 40 have a xanthene dye skeleton.

[0160] Fluorescent dye 41 is a fluorescent dye having a boron dipyrromethene dye skeleton (dye residue) containing the following structure. For example, in the substituent group "-Z-R1-R2-R3" of fluorescent dye 41, Z is -C2H4-, R1 is -C(=O)NH-, R2 is -C3H6-, and R3 is -N(CH3)2.

[0161]

[0162] [Table 1-5]

[0163]

[0164] [Table 1-6]

[0165]

[0166] [Table 1-7]

[0167]

[0168] [Table 1-8]

[0169]

[0170] Among the exemplified fluorescent dyes, fluorescent dyes 42 to 59 each have a phthalocyanine dye skeleton. Fluorescent dye 60 has a diketopyrrolopyrrole dye skeleton. Fluorescent dye 61 has a xanthene dye skeleton. Fluorescent dye 62 has a cyanine dye skeleton. Fluorescent dye 63 has a boron dipyrromethene dye skeleton.

[0171] Although not particularly limited, in one embodiment, the fluorescent labeling agent is preferably a fluorescent dye containing a phthalocyanine dye skeleton from the viewpoint of stability such as durability.

[0172] Example

[0173] Hereinafter, the present invention will be described based on Examples, but the present invention is not limited thereto. In the Examples, "parts" means "parts by mass."

[0174] (Quality Analysis)

[0175] The analysis was performed using a mass spectrometer (TOF-MS: Autoflex II manufactured by Bruker Daltonics).

[0176] Fluorescent pigments

[0177] [Production Example 1]

[0178] <Method for producing compound A-1>

[0179] Ammonia gas was introduced into a solution of 50 parts of quinoline and 1 part of anhydrous aluminum chloride, and 5 parts of 3-ethoxyphthalonitrile were added. The mixture was reacted at 180°C for 7 hours. After the reaction solution was cooled to room temperature, 200 parts of methanol and 200 parts of a 10% aqueous hydrochloric acid solution were added. The precipitated solid was then filtered and washed with 200 parts of water. The washed solid was dried at 80°C to obtain 4.7 parts of Compound A-1 (yield 88.4%) shown in Table 2.

[0180] [Manufacturing Examples 2 to 11]

[0181] <Methods for producing compounds A-2 to A-10>

[0182] Compounds A-2 to A-10 shown in Table 2 were produced in the same manner as in the production of Compound A-1, except that the 3-ethoxyphthalonitrile and anhydrous aluminum chloride used in the production method of Compound A-1 were replaced with the phthalonitrile derivatives and metal sources shown in Table 2. The phthalonitrile derivatives and metal sources were used in the same molar amounts as those of 3-ethoxyphthalonitrile and anhydrous aluminum chloride used in the production of Compound A-1.

[0183] [Table 2-1]

[0184]

[0185] [Table 2-2]

[0186]

[0187] [Production Example 11]

[0188] <Method for producing compound B-1>

[0189] To a solution obtained by dissolving 3 parts of compound A-1 in 10 parts of N-methyl-2-pyrrolidone (NMP), an aqueous solution obtained by dissolving 0.45 parts of potassium hydroxide in 1 part of water was added. These were reacted at 110°C for 7 hours. After the reaction solution was cooled to room temperature, 100 parts of water was added. Subsequently, the precipitated solid was filtered and washed with 100 parts of water. The washed solid was dried at 80°C to obtain 2.9 parts of compound B-1 shown in Table 3 (yield 99.2%).

[0190] [Manufacturing Examples 12 to 15]

[0191] <Methods for producing compounds B-2 to B-5>

[0192] Compounds B-2 to B-5 shown in Table 3 were prepared in the same manner as in the preparation of Compound B-1, except that Compound A-1 used in the preparation of Compound B-1 was replaced with Compound A shown in Table 3. Compound A was used in the same molar amount as Compound A-1 in the preparation of Compound B-1.

[0193] [Table 3]

[0194]

[0195]

[0196] [Production Example 16]

[0197] <Method for producing compound C-1>

[0198] Ammonia gas was introduced into a solution of 50 parts of quinoline and 1 part of anhydrous aluminum chloride, and 3.8 parts of 3-ethoxyphthalonitrile and 1.1 parts of 4-fluorophthalonitrile were added. These were reacted at 180°C for 7 hours. After the reaction solution was cooled to room temperature, 200 parts of methanol and 200 parts of a 10% aqueous hydrochloric acid solution were added. Subsequently, the precipitated solid was filtered out and washed with 200 parts of water. The washed solid (crude product) was refined using a medium-pressure fractionating liquid chromatograph (Smart Flash AKROS manufactured by Yamazen). The refined product obtained was dried at 80°C to obtain 1.6 parts of compound C-1 shown in Table 4 (yield 30.5%).

[0199] [Production Examples 17 to 19]

[0200] <Methods for producing compounds C-2 to C-4>

[0201] Phthalocyanines C-2 to C-4 shown in Table 4 were produced in the same manner as in the production of Compound C-1, except that the anhydrous aluminum chloride used in the production method of Compound C-1 was replaced with the metal sources shown in Table 4. The metal sources were used in the same molar amount as the anhydrous aluminum chloride used in the production of Compound C-1.

[0202] [Table 4]

[0203]

[0204]

[0205] [Example 1]

[0206] <Method for producing fluorescent dye 1>

[0207] 1 part of compound B-1 and 0.6 parts of 3-aminopropyldimethylethoxysilane were dissolved in pyridine, and the solution was refluxed at 115 ° C for 3 hours to obtain a reaction solution. After removing pyridine from the reaction solution using an evaporator, a mixed solution of 10 parts of ethanol and 50 parts of water was added. Then, the precipitated solid was filtered and washed with 50 parts of water. The washed solid was dried at 80 ° C to obtain 0.39 parts of the fluorescent dye 1 shown in Table 1 (yield 33.7%). As a result of mass analysis, a molecular ion peak was detected at m / z = 848.64 (theoretical value 847.99), which was identified as having the structure of the fluorescent dye 1 shown in Table 1.

[0208] [Example 2 to Example 5]

[0209] <Methods for producing fluorescent dyes 2 to 5>

[0210] Fluorescent pigments 2 to 5 shown in Table 1 were produced in the same manner as for the production of fluorescent pigment 1, except that compound B-1 used in the production method of fluorescent pigment 1 was replaced with compound B shown in Table 5. Compound B was used in the same molar amount as compound B-1 in the production of fluorescent pigment 1. The structures of the obtained fluorescent pigments 2 to 5 were identified by mass spectrometry and confirmed to have the structures shown in Table 1. Table 11 shows the results of mass spectrometry analysis.

[0211] [Table 5]

[0212] Fluorescent pigments Compound B Example 1 Fluorescent pigment 1 B-1 Example 2 Fluorescent pigment 2 B-2 Example 3 Fluorescent pigment 3 B-3 Example 4 Fluorochrome 4 B-4 Example 5 Fluorescent pigment 5 B-5

[0213] [Example 6]

[0214] <Method for producing fluorescent dye 6>

[0215] 0.7 parts of compound A-1 and 0.4 parts of 4-(3-aminopropyl)benzenesulfonic acid were dissolved in 50 parts of dimethyl sulfoxide, and 0.3 parts of 1,8-diazabicyclo[5.4.0]-7-undecene were added, and these were reacted at 90°C for 5 hours. After the reaction solution was cooled to room temperature, 100 parts of water and 10 parts of salt were added. Then, the precipitated solid was filtered and washed with 50 parts of water. The washed solid was dried at 80°C to obtain 0.36 parts of the fluorescent dye 6 shown in Table 1 (yield 41.6%). As a result of mass analysis, a molecular ion peak was detected at m / z=916.57 (theoretical value 915.96), which was identified as having the structure of the fluorescent dye 6 shown in Table 1.

[0216] [Example 7 to Example 14]

[0217] <Methods for producing fluorescent dyes 7 to 14>

[0218] Fluorescent pigments 7 to 14 shown in Table 1 were produced in the same manner as for the production of fluorescent pigment 6, except that Compound A-1 and 4-(3-aminopropyl)benzenesulfonic acid used in the production method of fluorescent pigment 6 were replaced with Compound A and the acidic compound shown in Table 6. Compound A and the acidic compound were used in the same molar amounts as Compound A-1 and 4-(3-aminopropyl)benzenesulfonic acid used in the production of fluorescent pigment 6. The structures of the obtained fluorescent pigments 7 to 14 were identified by mass spectrometry and confirmed to have the structures shown in Table 1. Table 11 shows the results of mass spectrometry analysis.

[0219] [Table 6]

[0220]

[0221] [Example 15]

[0222] <Method for producing fluorescent dye 15>

[0223] 0.5 parts of compound A-1 and 0.29 parts of (2-carboxyethyl)phenylphosphinic acid were dissolved in 20 parts of dimethyl sulfoxide, and the solution was reacted at 80°C for 8 hours. After the reaction solution was cooled to room temperature, 50 parts of water and 10 parts of salt were added. Then, the precipitated solid was filtered and washed with 50 parts of water. The washed solid was dried at 80°C to obtain 0.46 parts of the fluorescent dye 15 shown in Table 1 (yield 74.4%). As a result of mass analysis, a molecular ion peak was detected at m / z=929.46 (theoretical value 928.88), which was identified as having the structure of the fluorescent dye 15 shown in Table 1.

[0224] [Example 16 to Example 18]

[0225] <Methods for producing fluorescent dyes 16 to 18>

[0226] Fluorescent pigments 16 to 18 shown in Table 1 were produced in the same manner as for the production of fluorescent pigment 15, except that Compound A-1 and (2-carboxyethyl)phenylphosphinic acid used in the production method of fluorescent pigment 15 were replaced with Compound A and the acidic compound shown in Table 7. Compound A and the acidic compound were used in the same molar amounts as Compound A-1 and (2-carboxyethyl)phenylphosphinic acid used in the production of fluorescent pigment 15. The structures of the obtained fluorescent pigments 16 to 18 were identified by mass spectrometry and confirmed to have the structures shown in Table 1. Table 11 shows the results of mass spectrometry analysis.

[0227] [Table 7]

[0228]

[0229] [Example 19]

[0230] <Method for producing fluorescent dye 19>

[0231] 0.5 parts of fluorescent pigment 1, 0.8 parts of iodomethane, and 0.8 parts of potassium carbonate were dissolved in 50 parts of tetrahydrofuran, and the solution was reacted at 25 ° C for 5 hours. After removing tetrahydrofuran from the reaction solution using an evaporator, 20 parts of tetrahydrofuran and 60 parts of water were added. Then, the precipitated solid was filtered and washed with 60 parts of water. The washed solid was dried at 80 ° C to obtain 0.21 parts of the fluorescent pigment 19 shown in Table 1 (yield 33.3%). As a result of mass analysis, a molecular ion peak was detected at m / z (positive) = 892.25 (theoretical value 891.08), which was identified as having the structure of the fluorescent pigment 19 shown in Table 1.

[0232] [Example 20 to Example 23]

[0233] <Methods for producing fluorescent dyes 20 to 23>

[0234] Fluorescent pigments 20 to 23 shown in Table 1 were produced in the same manner as for producing fluorescent pigment 19, except that the iodinated compounds and amines shown in Table 8 were used instead of the methyl iodide and fluorescent pigment 1 used in the production method of fluorescent pigment 19. The iodinated compounds and amines were used in the same molar amounts as those of methyl iodide and fluorescent pigment 1 in the production of fluorescent pigment 19. The structures of the obtained fluorescent pigments 20 to 23 were identified by mass spectrometry and confirmed to have the structures shown in Table 1. Table 11 shows the results of mass spectrometry analysis.

[0235] [Table 8]

[0236]

[0237] [Example 24]

[0238] <Method for producing fluorescent dye 24>

[0239] 0.06 parts of fluorescent dye 1 and 0.007 parts of succinic anhydride were dissolved in 5 parts of N-methyl-2-pyrrolidone (NMP), and the solution was reacted at 90°C for 4 hours. After removing NMP from the reaction solution using a centrifugal evaporator, 5 parts of water were added. Then, the precipitated solid was filtered and washed with 5 parts of water. The washed solid was dried at 80°C to obtain 0.041 parts of fluorescent dye 24 shown in Table 1 (yield 61.1%). As a result of mass analysis, a molecular ion peak was detected at m / z=949.07 (theoretical value 948.06), which was identified as having the structure of fluorescent dye 24 shown in Table 1.

[0240] [Example 25 to Example 30]

[0241] <Methods for producing fluorescent dyes 25 to 30>

[0242] Fluorescent pigments 25 to 30 shown in Table 1 were produced in the same manner as for producing fluorescent pigment 24, except that succinic anhydride and fluorescent pigment 1 used in the production method of fluorescent pigment 24 were replaced with succinic anhydride derivatives and amines shown in Table 9. The succinic anhydride derivatives and amines were used in the same molar amounts as those of succinic anhydride and fluorescent pigment 1 used in the production of fluorescent pigment 24. The structures of the obtained fluorescent pigments 25 to 30 were identified by mass spectrometry and confirmed to have the structures shown in Table 1. Table 11 shows the results of mass spectrometry analysis.

[0243] [Table 9]

[0244]

[0245]

[0246] [Example 31]

[0247] <Method for producing fluorescent dye 31>

[0248] 1.0 parts of compound C-1 and 0.6 parts of (2-carboxyethyl)phenylphosphinic acid were dissolved in 50 parts of dimethyl sulfoxide, and 0.4 parts of 1,8-diazabicyclo[5.4.0]-7-undecene were added, and the solution was reacted at 90°C for 8 hours. After the reaction solution was cooled to room temperature, 100 parts of water was added. Then, the precipitated solid was filtered out and washed with 50 parts of water. The obtained solid (crude product) was purified using a medium-pressure fractionating liquid chromatograph (Smart Flash AKROS manufactured by Yamazen). The obtained purified product was dried at 80°C to obtain 0.72 parts of the fluorescent dye 31 shown in Table 1 (yield 60.1%). As a result of mass analysis, a molecular ion peak was detected at m / z=901.46 (theoretical value 900.82), and it was identified as having the structure of the fluorescent dye 31 shown in Table 1.

[0249] [Example 32 to Example 37]

[0250] <Methods for producing fluorescent dyes 32 to 37>

[0251] Fluorescent pigments 32 to 37 shown in Table 1 were produced in the same manner as for producing fluorescent pigment 31, except that compound C-1 and (2-carboxyethyl)phenylphosphinic acid used in the production method of fluorescent pigment 31 were replaced with compound C and an acidic compound shown in Table 10. Compound C and the acidic compound were used in the same molar amounts as compound C-1 and (2-carboxyethyl)phenylphosphinic acid used in the production of fluorescent pigment 31. The structures of the obtained fluorescent pigments 32 to 37 were identified by mass spectrometry and confirmed to have the structures shown in Table 1. Table 11 shows the results of mass spectrometry analysis.

[0252] [Table 10]

[0253]

[0254]

[0255] [Example 38]

[0256] <Method for producing fluorescent dye 38>

[0257] 1.0 part of diketopyrrolopyrrole pigment, Pigment Red 255 (manufactured by Tokyo Chemical Industry), 0.6 part of 4-bromobutyric acid, and 0.1 part of sodium hydride (60%) were dissolved in 50 parts of N,N-dimethylformamide, and the solution was reacted at 90°C for 4 hours. After the reaction solution was cooled to room temperature, 100 parts of water were added. Then, the precipitated solid was filtered and washed with 50 parts of water. The washed solid (crude product) was purified using a medium-pressure fractionating liquid chromatograph (Smart Flash AKROS manufactured by Yamazen). The purified product was dried at 80°C to obtain 0.70 parts of the fluorescent pigment 38 shown in Table 1 (yield 53.9%). As a result of mass analysis, a molecular ion peak was detected at m / z = 375.28 (theoretical value 374.40), and it was identified as having the structure of the fluorescent dye 38 shown in Table 1.

[0258] [Example 39]

[0259] <Method for producing fluorescent dye 39>

[0260] 1.0 parts of 5-carboxyfluorescein (manufactured by Tokyo Chemical Industry), 0.3 parts of N,N-dimethyl-1,3-propylenediamine, and 0.1 parts of p-toluenesulfonic acid were dissolved in 50 parts of xylene, and the solution was reacted at 140 ° C for 24 hours. After the reaction solution was cooled to room temperature, xylene was removed from the reaction solution using an evaporator, and 50 parts of petroleum ether were added. Then, after removing the insoluble matter by suction filtration, the petroleum ether was removed using an evaporator to obtain a solid. The solid was dried at 80 ° C to obtain 0.57 parts of the fluorescent pigment 39 shown in Table 1 (yield 46.6%). As a result of mass analysis, a molecular ion peak was detected at m / z = 461.37 (theoretical value 460.49), which was identified as having the structure of the fluorescent pigment 39 shown in Table 1.

[0261] [Example 40]

[0262] <Method for producing fluorescent dye 40>

[0263] 1.0 parts of rhodamin B (manufactured by Tokyo Chemical Industry), 0.2 parts of N,N-dimethyl-1,3-propylenediamine, and 0.1 parts of p-toluenesulfonic acid were dissolved in 50 parts of xylene, and the solution was reacted at 140 ° C for 24 hours. After the reaction solution was cooled to room temperature, xylene was removed from the reaction solution using an evaporator, and 50 parts of petroleum ether were added. Then, after removing the insoluble matter by suction filtration, the petroleum ether was removed using an evaporator to obtain a solid. The solid was dried at 80 ° C to obtain 0.46 parts of the fluorescent dye 40 shown in Table 1 (yield 39.1%). As a result of mass analysis, a molecular ion peak was detected at m / z = 564.02 (theoretical value 563.18), which was identified as having the structure of the fluorescent dye 40 shown in Table 1.

[0264] [Example 41]

[0265] <Method for producing fluorescent dye 41>

[0266] 1.0 parts of BODIPY pigment, namely BDP FL (manufactured by Tokyo Chemical Industry), and 0.3 parts of N,N-dimethyl-1,3-propylenediamine were dissolved in 50 parts of xylene, and the solution was reacted at 140°C for 24 hours. After the reaction solution was cooled to room temperature, xylene was removed from the reaction solution using an evaporator, and 50 parts of petroleum ether were added. Subsequently, after removing the insoluble matter by suction filtration, the petroleum ether was removed using an evaporator to obtain a solid. The solid was dried at 80°C to obtain 0.38 parts of the fluorescent pigment 41 shown in Table 1 (yield 29.3%). As a result of mass analysis, a molecular ion peak was detected at m / z=379.11 (theoretical value 378.27), which was identified as having the structure of the fluorescent pigment 41 shown in Table 1.

[0267] [Table 11]

[0268]

[0269] [Production Example 20]

[0270] <Method for producing compound A-11>

[0271] Ammonia gas was introduced into a solution of 50 parts quinoline and 1 part anhydrous aluminum chloride, followed by the addition of 5 parts 3,6-bis(phenylthio)phthalonitrile. The mixture was reacted at 180°C for 7 hours. After cooling the reaction solution to room temperature, 200 parts methanol and 200 parts 10% aqueous hydrochloric acid were added. The precipitated solid was then filtered and washed with 200 parts water. The washed solid was dried at 80°C to obtain Compound A-11 (yield 72.8%) shown in Table 12.

[0272] [Manufacturing Example 21, Manufacturing Example 22]

[0273] <Methods for producing Compound A-12 and Compound A-13>

[0274] Compounds A-12 and A-13 shown in Table 12 were produced in the same manner as in the production of Compound A-11, except that the 3,6-bis(phenylthio)phthalonitrile used in the production method of Compound A-11 was replaced with the phthalonitrile derivatives shown in Table 12. The phthalonitrile derivatives were used in the same molar amount as 3,6-bis(phenylthio)phthalonitrile in the production of Compound A-11.

[0275] [Table 12]

[0276]

[0277] [Example 42]

[0278] <Method for producing fluorescent dye 42>

[0279] 0.7 parts of compound A-1 and 0.4 parts of 1,2-ethylenediphosphonic acid were dissolved in 50 parts of dimethyl sulfoxide, and 0.3 parts of 1,8-diazabicyclo[5.4.0]-7-undecene were added, and the solution was reacted at 90°C for 5 hours. After the reaction solution was cooled to room temperature, 100 parts of water and 10 parts of salt were added. Then, the precipitated solid was filtered and washed with 50 parts of water. The washed solid was dried at 80°C to obtain 0.42 parts of the fluorescent dye 42 shown in Table 1 (yield 50.6%). As a result of mass analysis, a molecular ion peak was detected at m / z=905.35 (theoretical value 905.21), which was identified as having the structure of the fluorescent dye 42 shown in Table 1.

[0280] [Example 43 to Example 55]

[0281] <Methods for producing fluorescent dyes 43 to 55>

[0282] Fluorescent pigments 43 to 55, shown in Table 1, were produced in the same manner as for the production of fluorescent pigment 6, except that Compound A-1 and 1,2-ethylenediphosphonic acid used in the production method of fluorescent pigment 1 were replaced with Compound A and the axial ligand shown in Table 13. Compound A and the axial ligand were used in the same molar amounts as Compound A-1 and 1,2-ethylenediphosphonic acid used in the production of fluorescent pigment 42. The structures of the obtained fluorescent pigments 43 to 55 were identified by analysis using a mass spectrometer, confirming that they had the structures shown in Table 1. Table 13 shows the results of the mass spectrometry analysis.

[0283] [Table 13-1]

[0284]

[0285] [Table 13-2]

[0286]

[0287] [Table 13-3]

[0288]

[0289]

[0290] [Production Example 23]

[0291] <Production of Compound D-1>

[0292] To 200 parts of sulfolane and 15.7 parts of 1,8-diazabicyclo[5,4,0]-7-undecene (DBU) were added 5 parts of 4-butylthio-1,3-diiminoisoindoline and 8.8 parts of silicon tetrachloride. The mixture was heated and stirred at 160°C to 170°C for 8 hours. The reaction mixture was then cooled to room temperature (25°C) and 200 parts of methanol was added. The resulting precipitate (solid) was filtered, washed with a methanol:water (mass ratio 4:1) solution, and dried to obtain 2.6 parts of compound D-1 (63.6% yield) as shown in Table 14. As a result of mass spectrometry, a molecular ion peak was detected at m / z = 751.65 (theoretical value 751.24), confirming that the compound had the structure of Compound D-1 shown in Table 14.

[0293] [Table 14]

[0294]

[0295] [Example 56]

[0296] <Method for producing fluorescent dye 56>

[0297] 1.0 parts of compound B-5 and 0.5 parts of 1,2-ethylenediphosphonic acid were dissolved in 50 parts of dimethyl sulfoxide, and 0.3 parts of 1,8-diazabicyclo[5.4.0]-7-undecene were added, and these were reacted at 90°C for 5 hours. After the reaction solution was cooled to room temperature, 100 parts of water and 10 parts of salt were added. Then, the precipitated solid was filtered and washed with 50 parts of water. The washed solid was dried at 80°C to obtain 0.52 parts of the fluorescent dye 1 shown in Table 1 (yield 42.3%). As a result of mass analysis, a molecular ion peak was detected at m / z=923.47 (theoretical value 923.21), which was identified as having the structure of the fluorescent dye 56 shown in Table 1.

[0298] [Example 57]

[0299] <Method for producing fluorescent dye 57>

[0300] Fluorescent pigment 57 shown in Table 1 was produced in the same manner as for producing fluorescent pigment 56, except that compound B-5 used in the production method of fluorescent pigment 1 was replaced with compound D-1. Compound D-1 was used in the same molar amount as compound B-5 in the production of fluorescent pigment 56. The structure of the obtained fluorescent pigment 57 was identified by analysis using a mass spectrometer, and it was confirmed to have the structure shown in Table 1. Table 15 shows the results of mass spectrometry analysis.

[0301] [Table 15]

[0302]

[0303] [Example 58]

[0304] <Method for producing fluorescent dye 58>

[0305] 0.7 parts of compound C-3 and 0.7 parts of 1,2-hexamethylenediphosphonic acid were dissolved in 50 parts of dimethyl sulfoxide, and 0.3 parts of 1,8-diazabicyclo[5.4.0]-7-undecene were further added to the solution, and these were reacted at 90°C for 5 hours. After the reaction solution was cooled to room temperature, 100 parts of water and 10 parts of salt were added. Then, the precipitated solid was filtered and washed with 50 parts of water. The washed solid was dried at 80°C to obtain 0.39 parts of the fluorescent dye 1 shown in Table 1 (yield 42.3%). As a result of mass analysis, a molecular ion peak was detected at m / z=913.66 (theoretical value 913.24), which was identified as having the structure of the fluorescent dye 58 shown in Table 1.

[0306] [Example 59]

[0307] <Method for producing fluorescent dye 59>

[0308] Fluorescent Dye 59 shown in Table 1 was produced in the same manner as for the production of Fluorescent Dye 58, except that Compound C-3 and 1,2-hexamethylenediphosphonic acid used in the production method of Fluorescent Dye 58 were replaced with Compound C-4 and the ring substituents shown in Table 16. Compound C-4 was used in the same molar amount as Compound C-3 in the production of Fluorescent Dye 17. The structure of the obtained Fluorescent Dye 59 was identified by analysis using a mass spectrometer, and it was confirmed to have the structure shown in Table 1. Table 16 shows the results of the mass spectrometry analysis.

[0309] [Table 16]

[0310]

[0311] [Example 60]

[0312] <Method for producing fluorescent dye 60>

[0313] 1.0 part of diketopyrrolopyrrole pigment, Pigment Red 255 (Tokyo Chemical Industry), 0.6 part of 3-aminopropylphosphonic acid, and 0.1 part of sodium hydride (60% dispersion) were dissolved in 50 parts of N,N-dimethylformamide, and the solution was reacted at 90°C for 4 hours. After the reaction solution was cooled to room temperature, 100 parts of water were added. Then, the precipitated solid was filtered and washed with 50 parts of water. The washed solid (crude product) was refined using a medium-pressure fractionating liquid chromatograph (Smart Flash AKROS manufactured by Yamazen). The refined product obtained was dried at 80°C to obtain 0.65 parts of the fluorescent pigment 60 shown in Table 1 (yield 45.5%). As a result of mass analysis, a molecular ion peak was detected at m / z = 411.52 (theoretical value 411.10), and it was identified as having the structure of the fluorescent dye 60 shown in Table 1.

[0314] [Example 61]

[0315] <Method for producing fluorescent dye 61>

[0316] 1.0 parts of 5-carboxyfluorescein (Tokyo Chemical Industry), 0.7 parts of 3-aminopropylphosphonic acid, and 0.1 parts of p-toluenesulfonic acid were dissolved in 50 parts of xylene, and the solution was reacted at 140 ° C for 24 hours. After the reaction solution was cooled to room temperature, xylene was removed from the reaction solution using an evaporator, and 50 parts of petroleum ether were added. Then, after removing the insoluble matter by suction filtration, the petroleum ether was removed using an evaporator to obtain a solid. The solid was dried at 80 ° C to obtain 0.75 parts of the fluorescent pigment 61 shown in Table 1 (yield 51.2%). As a result of mass analysis, a molecular ion peak was detected at m / z = 551.02 (theoretical value 551.18), which was identified as having the structure of the fluorescent pigment 61 shown in Table 1.

[0317] [Example 62]

[0318] <Method for producing fluorescent dye 62>

[0319] 1.0 part of Cy5-NHS ester (Funakoshi), 0.5 part of 3-aminopropanol, and 0.5 part of triethylamine were dissolved in 50 parts of DMF, and the solution was reacted at room temperature for 12 hours. 50 parts of water were added to the reaction solution, and the precipitated precipitate (solid) was filtered, and the solid was washed with water. The washed solid was dried at 80°C to obtain 0.70 parts of the fluorescent pigment 62 shown in Table 1 (yield 86.4%). As a result of mass analysis, a molecular ion peak was detected at m / z=541.18 (theoretical value 541.36), which was identified as having the structure of the fluorescent pigment 62 shown in Table 1.

[0320] [Example 63]

[0321] <Method for producing fluorescent dye 63>

[0322] 1.0 parts of BODIPY pigment, namely BDP FL (manufactured by Tokyo Chemical Industry), and 0.3 parts of 3-aminopropanol were dissolved in 50 parts of xylene, and the solution was reacted at 140°C for 24 hours. After the reaction solution was cooled to room temperature, xylene was removed from the reaction solution using an evaporator, and 50 parts of petroleum ether were added. Subsequently, after removing the precipitated insoluble matter by suction filtration, petroleum ether was removed from the reaction solution using an evaporator to obtain a solid. The solid was dried at 80°C to obtain 0.64 parts of the fluorescent pigment 63 shown in Table 1 (yield 70.6%). As a result of mass analysis, a molecular ion peak was detected at m / z=350.01 (theoretical value 350.18), which was identified as having the structure of the fluorescent pigment 63 shown in Table 1.

[0323] [Comparative Example 1]

[0324] As comparative compound 1, compound A-1 was used.

[0325] [Comparative Example 2]

[0326] As comparative compound 2, compound A-9 was used.

[0327] In Comparative Examples 3 to 11 described below, Comparative Compounds 3 to 11 shown in Table 17 were produced.

[0328] [Table 17]

[0329]

[0330] [Comparative Example 3]

[0331] <Production Method of Comparative Compound 3>

[0332] 7.0 parts of aluminum chloride, 39 parts of urea, 0.2 parts of ammonium molybdate, and 25 parts of trimellitic anhydride were dissolved in 40 parts of N-methyl-2-pyrrolidone (NMP), and the solution was stirred at 139 ° C for 9 hours. After the reaction solution was cooled to room temperature, 100 parts of water was added. Then, the precipitated solid was filtered and washed with 50 parts of water. The washed solid was dried at 80 ° C to obtain 14.5 parts of comparative compound 3 shown in Table 17 (yield 59.3%). As a result of mass analysis, a molecular ion peak was detected at m / z = 751.84 (theoretical value 751.00), which was identified as having the structure of comparative compound 3 shown in Table 17.

[0333] [Comparative Example 4]

[0334] <Production Method of Comparative Compound 4>

[0335] Ammonia gas was introduced into a solution of 30 parts of quinoline and 0.7 parts of anhydrous aluminum chloride, and 1.5 parts of 3-ethoxyphthalonitrile and 2.1 parts of 4-octadecyloxyphthalonitrile were added, and the solution was reacted at 180°C for 7 hours. After the reaction solution was cooled to room temperature, 200 parts of methanol and 200 parts of a 10% aqueous hydrochloric acid solution were added. Subsequently, the precipitated solid was filtered out and washed with 200 parts of water. The washed solid (crude product) was refined using a medium-pressure fractionating liquid chromatograph (Smart Flash AKROS manufactured by Yamazen). The refined product obtained was dried at 80°C to obtain 0.36 parts of comparative compound 4 shown in Table 17 (yield 12.6%). As a result of mass analysis, a molecular ion peak was detected at m / z=976.44 (theoretical value 975.61), which was identified as having the structure of comparative compound 4 shown in Table 17.

[0336] [Comparative Example 5]

[0337] <Production Method of Comparative Compound 5>

[0338] 1.0 parts of compound A-9 and 0.35 parts of triphenylsilanol were dissolved in 20 parts of dimethyl sulfoxide, and the solution was reacted at 80 ° C for 8 hours. After the reaction solution was cooled to room temperature, 50 parts of water and 10 parts of salt were added. Then, the precipitated solid was filtered and washed with 50 parts of water. The washed solid was dried at 80 ° C to obtain 1.00 parts of comparative compound 5 shown in Table 17 (yield 80.5%). As a result of mass analysis, a molecular ion peak was detected at m / z = 1282.53 (theoretical value 1271.67), which was identified as having the structure of comparative compound 5 shown in Table 17.

[0339] [Comparative Examples 6 to 9]

[0340] <Methods for producing comparative compounds 6 to 9>

[0341] Comparative Compounds 6 to 9 shown in Table 17 were prepared in the same manner as for the preparation of Comparative Compound 5, except that Compound A-9 and triphenylsilanol used in the preparation of Comparative Compound 5 were replaced with the halogens and acidic compounds shown in Table 18. Halogens and acidic compounds were used in the same molar amounts as those used for Compound A-9 and triphenylsilanol in the preparation of Comparative Compound 5. The structures of the obtained Comparative Compounds 6 to 9 were identified by mass spectrometry and confirmed to have the structures shown in Table 17. Table 19 shows the results of mass spectrometry analysis.

[0342] [Table 18]

[0343]

[0344] [Comparative Example 10]

[0345] <Production Method of Comparative Compound 10>

[0346] 2.0 parts of compound A-9 and 1.0 parts of p-toluenesulfonic acid were dissolved in 50 parts of dimethyl sulfoxide, and 0.3 parts of 1,8-diazabicyclo[5.4.0]-7-undecene were added to the solution and reacted at 90°C for 5 hours. After the reaction solution was cooled to room temperature, 100 parts of water were added. Subsequently, the precipitated solid was filtered and washed with 50 parts of water. The washed solid was dried at 80°C to obtain 1.35 parts of comparative compound 10 (yield 60.0%) shown in Table 17. As a result of mass analysis, a molecular ion peak was detected at m / z=933.67 (theoretical value 932.74), which was identified as having the structure of comparative compound 10 shown in Table 17.

[0347] [Comparative Example 11]

[0348] <Production Method of Comparative Compound 11>

[0349] To a mixed solution of 9.2 parts of concentrated sulfuric acid and 5.5 parts of 25% fuming sulfuric acid was added 1 part of compound A-1, and the solution was heated and stirred at 50 ° C for 4 hours. After the reaction solution was cooled, it was added to 80 parts of ice, and the precipitated precipitate (solid) was filtered and separated. Furthermore, the solid after filtration and separation was suspended in 50 parts of tetrahydrofuran, and the precipitate was filtered and separated again. The solid after filtration and separation was washed with 50 parts of tetrahydrofuran, and the washed solid was dried to obtain 0.5 parts of crude product. The crude product was refined using a medium-pressure fractionating liquid chromatograph (Smart Flash AKROS manufactured by Yamazen) to obtain 0.2 parts of comparative compound 11 (yield 16.0%). As a result of mass analysis, a molecular ion peak was detected at m / z = 939.65 (theoretical value 940.75), which was identified as having the structure of comparative compound 11 shown in Table 17.

[0350] The results of mass spectrometric analysis of Comparative Compounds 3 to 11 produced in Comparative Examples 3 to 11 are shown.

[0351] [Table 19]

[0352] Theoretical value Measured value Comparative Compound 3 751.00 751.84 Comparative Compound 4 975.61 976.44 Comparative Compound 5 1271.67 1282.53 Comparative Compound 6 1127.54 1128.49 Comparative Compound 7 1169.62 1170.82 Comparative Compound 8 1231.45 1232.37 Comparative Compound 9 932.74 933.67 Comparative Compound 10 1167.46 1168.37 Comparative Compound 11 940.75 939.65

[0353] [Comparative Example 12]

[0354] A cyanine-based fluorescent dye (XenoLight DIR) (manufactured by Summit Pharma) was used as comparative compound 12. This compound corresponds to a conventional fluorescent labeling agent having a long-chain alkylene group and accumulates on phospholipids through hydrophobic interaction.

[0355]

[0356] [Comparative Example 13]

[0357] As comparative compound 13, Rhodamine B (manufactured by Tokyo Chemical Industry Co., Ltd.) was used.

[0358] <II> Pigment solution

[0359] [Example 64]

[0360] <Preparation of pigment solution 1>

[0361] 1.696 mg of fluorescent dye 1 was dissolved in 10 ml of dimethyl sulfoxide. The solution was filtered using a nylon membrane filter with a pore size of 0.2 μm, and then diluted 100-fold with dimethyl sulfoxide to prepare a dye solution 1 of fluorescent dye 1.

[0362] [Example 65 to Example 126]

[0363] <Preparation of Dye Solutions 2 to 63>

[0364] Dye solutions 2 to 63 were prepared in the same manner as in the preparation of dye solution 1, except that the fluorescent dye 1 and dimethyl sulfoxide used in the preparation of dye solution 1 were replaced with the fluorescent dyes and solvents shown in Table 20. Each fluorescent dye was used in the same molar amount as that of fluorescent dye 1, and the solvent was used in the same volume amount as that of dimethyl sulfoxide.

[0365] [Comparative Examples 12 to 24]

[0366] <Preparation of Dye Solutions 64 to 76>

[0367] Dye solutions 64 to 76 were prepared in the same manner as in the preparation of dye solution 1, except that the fluorescent dye 1 and dimethyl sulfoxide used in the preparation of dye solution 1 were replaced with the fluorescent dyes and solvents shown in Table 20. Each fluorescent dye was used in the same molar amount as that of fluorescent dye 1, and the solvent was used in the same volumetric amount as that of dimethyl sulfoxide.

[0368] [Table 20-1]

[0369] Pigment solution Fluorescent pigments solvent Example 64 Pigment solution 1 Fluorescent pigment 1 DMSO Example 65 Pigment solution 2 Fluorescent pigment 2 DMSO Example 66 Pigment solution 3 Fluorescent pigment 3 DMSO Example 67 Pigment solution 4 Fluorochrome 4 DMSO Example 68 Pigment solution 5 Fluorescent pigment 5 DMSO Example 69 Pigment solution 6 Fluorescent pigment 6 DMSO Example 70 Pigment solution 7 Fluorescent pigment 7 DMSO Example 71 Pigment solution 8 Fluorescent pigment 8 DMSO Example 72 Pigment solution 9 Fluorescent pigment 9 DMSO Example 73 Pigment solution 10 Fluorescent pigment 10 DMSO Example 74 Pigment solution 11 Fluorescent pigment 11 DMSO Example 75 Pigment solution 12 Fluorescent pigment 12 DMSO Example 76 Pigment solution 13 Fluorescent pigment 13 DMSO Example 77 Pigment solution 14 Fluorescent pigment 14 DMSO Example 78 Pigment solution 15 Fluorescent pigment 15 DMSO Example 79 Pigment solution 16 Fluorescent pigment 16 DMSO Example 80 Pigment solution 17 Fluorescent pigment 17 DMSO Example 81 Pigment solution 18 Fluorescent pigment 18 DMSO Example 82 Pigment solution 19 Fluorescent pigment 19 DMSO Example 83 Pigment solution 20 Fluorescent pigment 20 DMSO Example 84 Pigment solution 21 Fluorescent pigment 21 DMSO Example 85 Pigment solution 22 Fluorescent pigment 22 DMSO Example 86 Pigment solution 23 Fluorescent pigment 23 DMSO Example 87 Pigment solution 24 Fluorescent pigment 24 DMSO Example 88 Pigment solution 25 Fluorescent pigment 25 DMSO Example 89 Pigment solution 26 Fluorescent pigment 26 DMSO Example 90 Pigment solution 27 Fluorochrome 27 DMSO Example 91 Pigment solution 28 Fluorochrome 28 DMSO Example 92 Pigment solution 29 Fluorescent pigment 29 DMSO Example 93 Pigment solution 30 Fluorescent pigment 30 DMSO Example 94 Pigment solution 31 Fluorescent pigment 31 DMSO Example 95 Pigment solution 32 Fluorescent pigment 32 DMSO

[0370] [Table 20-2]

[0371] Pigment solution Fluorescent pigments solvent Example 96 Pigment solution 33 Fluorescent pigment 33 DMSO Example 97 Pigment solution 34 Fluorescent pigment 34 DMSO Example 98 Pigment solution 35 Fluorescent pigment 35 DMSO Example 99 Pigment solution 36 Fluorescent pigment 36 DMSO Example 100 Pigment solution 37 Fluorescent pigment 37 DMSO Example 101 Pigment solution 38 Fluorescent pigment 38 DMSO Example 102 Pigment solution 39 Fluorescent pigment 39 DMSO Example 103 Pigment solution 40 Fluorescent pigment 40 DMSO Example 104 Pigment solution 41 Fluorescent pigment 41 DMSO Example 105 Pigment solution 42 Fluorescent pigment 42 DMSO Example 106 Pigment solution 43 Fluorescent pigment 43 DMSO Example 107 Pigment solution 44 Fluorescent pigment 44 DMSO Example 108 Pigment solution 45 Fluorescent pigment 45 DMSO Example 109 Pigment solution 46 Fluorescent pigment 46 DMSO Example 110 Pigment solution 47 Fluorochrome 47 DMSO Example 111 Pigment solution 48 Fluorescent pigment 48 DMSO Example 112 Pigment solution 49 Fluorescent pigment 49 DMSO Example 113 Pigment solution 50 Fluorescent pigment 50 DMSO Example 114 Pigment solution 51 Fluorescent pigment 51 DMSO Example 115 Pigment solution 52 Fluorescent pigment 52 DMSO Example 116 Pigment solution 53 Fluorescent pigment 53 DMSO Example 117 Pigment solution 54 Fluorescent pigment 54 DMSO Example 118 Pigment solution 55 Fluorescent pigment 55 DMSO Example 119 Pigment solution 56 Fluorescent pigment 56 DMSO Example 120 Pigment solution 57 Fluorescent pigment 57 DMSO Example 121 Pigment solution 58 Fluorescent pigment 58 DMSO Example 122 Pigment solution 59 Fluorescent pigment 59 DMSO Example 123 Pigment solution 60 Fluorescent pigment 60 DMSO Example 124 Pigment solution 61 Fluorescent pigment 61 DMSO Example 125 Pigment solution 62 Fluorescent pigment 62 DMSO Example 126 Pigment solution 63 Fluorescent pigment 63 DMSO

[0372] [Table 20-3]

[0373] Pigment solution Fluorescent pigments solvent Comparative Example 12 Pigment solution 64 Comparative Compound 1 DMSO Comparative Example 13 Pigment solution 65 Comparative Compound 2 DMSO Comparative Example 14 Pigment solution 66 Comparative Compound 3 DMSO Comparative Example 15 Pigment solution 67 Comparative Compound 4 DMSO Comparative Example 16 Pigment solution 68 Comparative Compound 5 DMSO Comparative Example 17 Pigment solution 69 Comparative Compound 6 DMSO Comparative Example 18 Pigment Solution 70 Comparative Compound 7 DMSO Comparative Example 19 Pigment solution 71 Comparative Compound 8 DMSO Comparative Example 20 Pigment solution 72 Comparative Compound 9 DMSO Comparative Example 21 Pigment solution 73 Comparative Compound 10 DMSO Comparative Example 22 Pigment solution 74 Comparative Compound 11 DMSO Comparative Example 23 Pigment Solution 75 Comparative Compound 12 DMSO Comparative Example 24 Pigment solution 76 Comparative Compound 13 water

[0374] <Evaluation of fluorescence intensity of dye solution>

[0375] Fluorescence spectra were measured for each pigment solution using a fluorescence spectrophotometer (manufactured by JASCO Corporation, FP-6500). Fluorescence intensity was then calculated by adding the obtained measured values to the fluorescence intensity within the range of fluorescence wavelengths shown in Table 22. The excitation light used at this time had a wavelength corresponding to the maximum absorption wavelength on the longest wavelength side of the pigment.

[0376] <III> Fluorescent marking agent

[0377] [Example 127]

[0378] <Preparation of Fluorescent Labeling Agent 1>

[0379] 1.696 mg of fluorescent dye 1 was dissolved in 10 ml of dimethyl sulfoxide, filtered through a nylon membrane filter with a pore size of 0.2 μm, and then diluted 100-fold with Roswell Park Memorial Institute (RPMI) 1640 medium to prepare a fluorescent labeling agent 1 of fluorescent dye 1.

[0380] [Example 128 to Example 189]

[0381] <Preparation of Fluorescent Labeling Agents 2 to 63>

[0382] Fluorescent Labeling Agents 2 to 63 were prepared in the same manner as for Fluorescent Labeling Agent 1, except that the fluorescent dye 1 and dimethyl sulfoxide used in the preparation of Fluorescent Labeling Agent 1 were replaced with the fluorescent dyes and solvents shown in Table 21. Each fluorescent dye was used in the same molar amount as that of fluorescent dye 1, and the solvent was used in the same volumetric amount as that of dimethyl sulfoxide.

[0383] [Comparative Examples 25 to 37]

[0384] <Preparation of Fluorescent Labeling Agents 64 to 76>

[0385] Fluorescent labeling agents 64 to 76 were prepared in the same manner as in the preparation of fluorescent labeling agent 1, except that the fluorescent dye 1 and dimethyl sulfoxide used in the preparation of fluorescent labeling agent 1 were replaced with the fluorescent dyes and solvents shown in Table 21. Each fluorescent dye was used in the same molar amount as that of fluorescent dye 1, and the solvent was used in the same volumetric amount as that of dimethyl sulfoxide.

[0386] [Table 21-1]

[0387] Fluorescent markers Fluorescent pigments solvent Example 127 Fluorescent marker 1 Fluorescent pigment 1 DMSO Example 128 Fluorescent marker 2 Fluorescent pigment 2 DMSO Example 129 Fluorescent marker 3 Fluorescent pigment 3 DMSO Example 130 Fluorescent marker 4 Fluorochrome 4 DMSO Example 131 Fluorescent marker 5 Fluorescent pigment 5 DMSO Example 132 Fluorescent marker 6 Fluorescent pigment 6 DMSO Example 133 Fluorescent marker 7 Fluorescent pigment 7 DMSO Example 134 Fluorescent marker 8 Fluorescent pigment 8 DMSO Example 135 Fluorescent marker 9 Fluorescent pigment 9 DMSO Example 136 Fluorescent marker 10 Fluorescent pigment 10 DMSO Example 137 Fluorescent marker 11 Fluorescent pigment 11 DMSO Example 138 Fluorescent marker 12 Fluorescent pigment 12 DMSO Example 139 Fluorescent marker 13 Fluorescent pigment 13 DMSO Example 140 Fluorescent marker 14 Fluorescent pigment 14 DMSO Example 141 Fluorescent marker 15 Fluorescent pigment 15 DMSO Example 142 Fluorescent marker 16 Fluorescent pigment 16 DMSO Example 143 Fluorescent marker 17 Fluorescent pigment 17 DMSO Example 144 Fluorescent marker 18 Fluorescent pigment 18 DMSO Example 145 Fluorescent marker 19 Fluorescent pigment 19 DMSO Example 146 Fluorescent marker 20 Fluorescent pigment 20 DMSO Example 147 Fluorescent marker 21 Fluorescent pigment 21 DMSO Example 148 Fluorescent marker 22 Fluorescent pigment 22 DMSO Example 149 Fluorescent marker 23 Fluorescent pigment 23 DMSO Example 150 Fluorescent marker 24 Fluorescent pigment 24 DMSO Example 151 Fluorescent marker 25 Fluorescent pigment 25 DMSO Example 152 Fluorescent marker 26 Fluorescent pigment 26 DMSO Example 153 Fluorescent marker 27 Fluorochrome 27 DMSO Example 154 Fluorescent marker 28 Fluorochrome 28 DMSO Example 155 Fluorescent marker 29 Fluorescent pigment 29 DMSO Example 156 Fluorescent marker 30 Fluorescent pigment 30 DMSO Example 157 Fluorescent marker 31 Fluorescent pigment 31 DMSO Example 158 Fluorescent marker 32 Fluorescent pigment 32 DMSO

[0388] [Table 21-2]

[0389] Fluorescent markers Fluorescent pigments solvent Example 159 Fluorescent marker 33 Fluorescent pigment 33 DMSO Example 160 Fluorescent marker 34 Fluorescent pigment 34 DMSO Example 161 Fluorescent marker 35 Fluorescent pigment 35 DMSO Example 162 Fluorescent marker 36 Fluorescent pigment 36 DMSO Example 163 Fluorescent marker 37 Fluorescent pigment 37 DMSO Example 164 Fluorescent marker 38 Fluorescent pigment 38 DMSO Example 165 Fluorescent marker 39 Fluorescent pigment 39 DMSO Example 166 Fluorescent marker 40 Fluorescent pigment 40 DMSO Example 167 Fluorescent marker 41 Fluorescent pigment 41 DMSO Example 168 Fluorescent marker 42 Fluorescent pigment 42 DMSO Example 169 Fluorescent marker 43 Fluorescent pigment 43 DMSO Example 170 Fluorescent marker 44 Fluorescent pigment 44 DMSO Example 171 Fluorescent marker 45 Fluorescent pigment 45 DMSO Example 172 Fluorescent marker 46 Fluorescent pigment 46 DMSO Example 173 Fluorescent marker 47 Fluorochrome 47 DMSO Example 174 Fluorescent marker 48 Fluorescent pigment 48 DMSO Example 175 Fluorescent marker 49 Fluorescent pigment 49 DMSO Example 176 Fluorescent marker 50 Fluorescent pigment 50 DMSO Example 177 Fluorescent marker 51 Fluorescent pigment 51 DMSO Example 178 Fluorescent marker 52 Fluorescent pigment 52 DMSO Example 179 Fluorescent marker 53 Fluorescent pigment 53 DMSO Example 180 Fluorescent marker 54 Fluorescent pigment 54 DMSO Example 181 Fluorescent marker 55 Fluorescent pigment 55 DMSO Example 182 Fluorescent marker 56 Fluorescent pigment 56 DMSO Example 183 Fluorescent marker 57 Fluorescent pigment 57 DMSO Example 184 Fluorescent marker 58 Fluorescent pigment 58 DMSO Example 185 Fluorescent marker 59 Fluorescent pigment 59 DMSO Example 186 Fluorescent marker 60 Fluorescent pigment 60 DMSO Example 187 Fluorescent marker 61 Fluorescent pigment 61 DMSO Example 188 Fluorescent marker 62 Fluorescent pigment 62 DMSO Example 189 Fluorescent marker 63 Fluorescent pigment 63 DMSO

[0390] [Table 21-3]

[0391] Fluorescent markers Fluorescent pigments solvent Comparative Example 25 Pigment solution 64 Comparative Compound 1 DMSO Comparative Example 26 Pigment solution 65 Comparative Compound 2 DMSO Comparative Example 27 Pigment solution 66 Comparative Compound 3 DMSO Comparative Example 28 Pigment solution 67 Comparative Compound 4 DMSO Comparative Example 29 Pigment solution 68 Comparative Compound 5 DMSO Comparative Example 30 Pigment solution 69 Comparative Compound 6 DMSO Comparative Example 31 Pigment Solution 70 Comparative Compound 7 DMSO Comparative Example 32 Pigment solution 71 Comparative Compound 8 DMSO Comparative Example 33 Pigment solution 72 Comparative Compound 9 DMSO Comparative Example 34 Pigment solution 73 Comparative Compound 10 DMSO Comparative Example 35 Pigment solution 74 Comparative Compound 11 DMSO Comparative Example 36 Pigment Solution 75 Comparative Compound 12 DMSO Comparative Example 37 Pigment solution 76 Comparative Compound 13 RPMI1640 medium

[0392] <Evaluation of cytotoxicity of fluorescent labeling agents>

[0393] Human epithelial cancer cells A431 were seeded into 96-well plates (1×10 4 The A431 cells were then cultured for 24 hours in an incubator (37°C, 5% CO₂ in air, humidified) using RPMI1640 medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin. RPMI1640 medium manufactured by Sigma-Aldrich was used.

[0394] After the culture, the culture medium was removed, and the fluorescent markers prepared in Examples 83 to 123 and Comparative Examples 25 to 37 and RPMI1640 culture medium (DMSO culture medium solution) containing 1% dimethyl sulfoxide were added. After these were allowed to stand in an incubator for 1 hour, they were washed with RPMI1640 culture medium. 10 μL of a cell counting kit (Cell Counting Kit)-8 (manufactured by Tongren Chemical) was added to each well and allowed to stand in an incubator (37°C, air containing 5% CO2, humidified environment) for 1 hour. Subsequently, the absorbance at 450 nm was measured using a disk reader (manufactured by TECAN, SPARK).

[0395] The relative value of the absorbance of each fluorescent marker when the absorbance of the well to which the DMSO culture medium solution was added was calculated and evaluated based on the following benchmark. It can be said that if it is evaluated as "P", cytotoxicity will not be shown. In addition, when calculating the relative value of the absorbance of the fluorescent marker, the value obtained by subtracting the absorbance of the cell counting kit-8 (manufactured by Tongren Chemical) before addition from the measured absorbance is used. The evaluation results are shown in Table 22.

[0396] (Evaluation Criteria)

[0397] P (pass): 0.8 or above

[0398] F (failure): less than 0.8

[0399] <Evaluation of Fluorescence Intensity of Fluorescent Labeling Agents>

[0400] Human epithelial cancer cells A431 were seeded into 96-well plates (1×10 4 Then, the A431 cells were cultured in RPMI1640 medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin in an incubator (37° C., air containing 5% CO 2 , humidified environment) for 24 hours.

[0401] After the incubation, the culture medium was removed and the fluorescent labeling agents prepared in Examples 127 to 167 and Comparative Examples 25 to 37 were added. The cells were then allowed to stand in an incubator for 1 hour. The cells were then washed with RPMI1640 culture medium. Fluorescence intensity was evaluated within the fluorescence wavelength range shown in Table 22 using a plate reader (SPARK, manufactured by TECAN).

[0402] Figure 1 The following table shows the results of fluorescence intensity evaluation of fluorescent labeling agent 1, fluorescent labeling agent 15, fluorescent labeling agent 19, fluorescent labeling agent 24, fluorescent labeling agent 25, fluorescent labeling agent 68, and fluorescent labeling agent 75. It was confirmed that fluorescent labeling agent 1, fluorescent labeling agent 15, fluorescent labeling agent 19, fluorescent labeling agent 24, and fluorescent labeling agent 25 (Example), which are embodiments of the present invention, exhibited higher fluorescence intensities than fluorescent labeling agent 68 and fluorescent labeling agent 75 (Comparative Example), which were prepared using comparative compounds.

[0403] <Evaluation of the Phospholipid Aggregation of Fluorescent Dyes>

[0404] The phospholipid aggregation properties of each pigment were calculated using formula (1) based on the integrated fluorescence intensity values obtained from the fluorescence spectra of the pigment solutions shown in Table 22 and the fluorescence intensity obtained from the fluorescence intensity of the fluorescent labeling agent. The relative values of the phospholipid aggregation properties of each fluorescent labeling agent were calculated, with the phospholipid aggregation property of the comparative compound 12 being set to 1, and evaluated based on the following criteria. When the evaluation was 3 or higher, it can be said that the phospholipid aggregation properties of each fluorescent pigment were good.

[0405] (Evaluation Criteria)

[0406] 4: Phospholipid aggregation is 4 or more

[0407] 3: Phospholipid aggregation is 2 or more and less than 4

[0408] 2: Phospholipid aggregation is 1 or more and less than 2

[0409] 1: Phospholipid aggregation is less than 1

[0410]

[0411] Table 22 shows the evaluation results of phospholipid aggregation.

[0412] It was confirmed that the fluorescent labeling agent (Example) according to the embodiment of the present invention exhibited higher phospholipid aggregating properties than the fluorescent labeling agent (Comparative Example) prepared using the comparative compound.

[0413] <Evaluation of cell visibility>

[0414] Human epithelial cancer cells A431 were seeded into 96-well plates (1×10 4 The A431 cells were cultured in RPMI1640 medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin in an incubator (37° C., air containing 5% CO 2 , humidified environment) for 24 hours.

[0415] After the culture, the medium was removed, and the fluorescent labeling agents prepared in Examples 127 to 189 and Comparative Examples 25 to 37 were added, and the cells were left to stand in an incubator for 1 hour. Subsequently, the cells were washed with RPMI1640 medium.

[0416] Using a fluorescence microscope (Keyence BZ-X800) equipped with excitation filters and fluorescence filters of appropriate wavelengths, dark field images and fluorescent images of the cells were observed and evaluated based on the following criteria. The evaluation results are shown in Table 22.

[0417] (Evaluation Criteria)

[0418] P (Pass): Clear

[0419] F (Fail): Unclear [Table 22-1]

[0420]

[0421] [Table 22-2]

[0422]

[0423] [Table 22-3]

[0424]

[0425] The results of the evaluation of the visibility of cells labeled with the fluorescent labeling agent 1, the fluorescent labeling agent 15, the fluorescent labeling agent 19, the fluorescent labeling agent 24, the fluorescent labeling agent 25, the fluorescent labeling agent 68, and the fluorescent labeling agent 75 are shown in order. Figures 2 to 8 (Magnification: 10x, fluorescence acquisition time: 1 second).

[0426] The results of the evaluation of the visibility of cells labeled with the fluorescent labeling agent 42 and the fluorescent labeling agent 53 are shown in order. Figure 9 and Figure 10 (Magnification: 40x, fluorescence acquisition time: 1 second).

[0427] The corresponding comparative example Figure 7 and Figure 8 and corresponding to the embodiment Figures 2 to 6 、 Figure 9 and Figure 10 As shown in the comparison of the embodiment of the present invention, it is observed that the fluorescent marker according to the embodiment of the present invention exhibits a higher fluorescence intensity derived from a specific substituent. Thus, it can be seen that the fluorescent marker according to the embodiment of the present invention (Example) has excellent cell aggregation compared to the comparative compound, thereby obtaining better visibility. Based on the above, it is clear that the fluorescent marker according to the embodiment of the present invention has excellent properties as a fluorescent marker.

Claims

1. A fluorescent labeling agent comprising a fluorescent dye, wherein the fluorescent dye has a fluorescent dye residue and a substituent bonded to the fluorescent dye residue in its structure. in, The residue of the fluorescent pigment is a residue of a phthalocyanine pigment, a residue of a diketopyrrolopyrrole pigment, a residue of a xanthene pigment, a residue of a boron dipyrromethene pigment, or a residue of a cyanine pigment, and The fluorescent pigment is a compound selected from the group consisting of the following compounds 1 to 63, The fluorescent labeling agent according to claim 1 , which is a phospholipid aggregation type fluorescent labeling agent.

3. A compound having a phthalocyanine residue and a substituent bonded to the phthalocyanine residue in its structure, and being a compound selected from the group consisting of Compounds 1 to 37 and Compounds 42 to 59 below,

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