A fluorescent dye compound, and a preparation method and application thereof

Abstract

The application belongs to the technical field of fluorescent dyes, and particularly relates to a fluorescent dye compound, a preparation method and application thereof. The fluorescent dye compound provided by the application has a Cy-R structure, wherein R is a group obtained by reacting with an aldehyde group. By introducing the R group on the basis of the Cy structure, the photophysical performance of the compound is greatly improved compared with the compound without R, so that the saturation loss laser power of the dye can be reduced, and the saturation loss power in stimulated emission loss super-resolution imaging is reduced, the permeability and compatibility of the compound for living cells are enhanced, the compound can be better used for stimulated emission loss super-resolution imaging of living cells, the application range of the dye in biological imaging is improved, the compound with Cy-R has good light bleaching resistance and light stability, and the compound can be applied to super-resolution imaging of living cells to characterize more internal details of cells.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of fluorescent dye technology and biological imaging, and particularly relates to a fluorescent dye compound and a preparation method and application thereof. BACKGROUND

[0002] With the development of cell biology, molecular imaging diagnostic techniques have gradually emerged, including confocal fluorescence imaging microscopy, fluorescence spectrum / lifetime instrument and other technologies. In recent years, super-resolution imaging methods have been continuously developed, which can break through the limit of optical diffraction limit and realize high-resolution imaging, revealing microstructures smaller than 200 nm. Fluorescent dyes, as a key component of fluorescence imaging and super-resolution fluorescence imaging, play an important role in the development of imaging diagnostic analysis technology. Most of the existing super-resolution imaging dyes can only emit single-color fluorescence. However, dyes capable of simultaneously labeling and detecting multiple molecules, organisms or multiple organelles are more practical. With the progress of scientific research and technology, more and more dyes with multi-color luminescence and low energy loss characteristics have been developed, but the development of fluorescent dyes with both of these two characteristics and capable of realizing super-resolution imaging of living cells still has room for improvement and innovation. The multi-color luminescence performance of fluorescent dyes depends on the design of their molecular structure and emission energy level. By adjusting the structure and chemical functional groups of the dye, multiple emission energy levels and adjustable emission wavelengths can be achieved, thereby producing different colors of luminescence, which is of great significance for multi-color labeling and imaging in analysis and detection applications. At the same time, low energy loss is another key element in dye design. High-power loss laser can exacerbate the photobleaching phenomenon of fluorescent materials, and cause greater damage to biological tissues, which leads to certain technical bottlenecks in stimulated emission depletion microscopy (STED) super-resolution imaging of living cells. It is of great significance to develop dyes that can emit strong fluorescence under low-energy excitation, thereby realizing long-term fluorescence imaging, and to study super-resolution imaging of living cells for exploring real biological structures and processes. Therefore, it is of great significance to research and develop more effective fluorescent materials and their preparation techniques for super-resolution imaging of living cells.

[0003] Commonly used fluorescence imaging materials include several categories such as organic monomolecular fluorescent dyes, organic polymer fluorescent nanoballs, and inorganic nanoluminescent materials. Compared with organic polymer nanoballs and inorganic nanoluminescent materials, organic monomolecular fluorescent dyes have obvious advantages and are concerned, and are a large category of materials and are widely used. The dye types are various, and the commonly used organic fluorescent dyes have a wide range of emission wavelengths, which can be divided into blue (coumarin, naphthalimide), green (BODIPY, fluorescein), red (rhodamine, cyanine fluorescent dye), and the like. Due to the advantages such as suitable absorption and emission properties, low cytotoxicity, and the like, these dyes have a wide application in life science analysis. Among these fluorescent dyes, the fluorescent dyes with red emission wavelength such as cyanine fluorescent dyes have absorption, emission wavelength, large molar absorption coefficient (×10 5 ), suitable fluorescence quantum yield, and low cytotoxicity, and are widely used in many aspects such as single molecule level in cells, small animal living bodies, and the like, and have obtained good development and application in super-resolution imaging. However, due to the needs of biological imaging research, fluorescent dyes with different emission wavelengths can be used for multicolor labeling of cells, high-contrast imaging research of different biological functional regions, and the like, and therefore it is necessary to continue to develop luminescent fluorescent dyes suitable for live cell imaging to meet the application needs.

[0004] In summary, in the related art, the types of super-resolution imaging dyes are few, and the related dye biomolecule targeting labeling is difficult, the imaging spatial resolution of live cell substructures is low, and the observation of the dynamic change process of live cells is difficult. SUMMARY

[0005] The purpose of the present application is to overcome the problems in the related art that the types of super-resolution imaging dyes are few, and the related dye biomolecule targeting labeling is difficult, the imaging spatial resolution of live cell substructures is low, and the observation of the dynamic change process of live cells is difficult, and to provide a fluorescent dye compound, a derivative thereof, and a preparation method and application thereof.

[0006] A fluorescent dye compound has the structure shown in the following formula B:

[0007] Cy-R formula B;

[0008] wherein, wherein R is a group obtained by reacting with an aldehyde group; Cy- is selected from one of the following structures:

[0009]

[0010] R1, R2, R3are selected from any one of hydrogen, halogen, benzyl, ester group, amide group, carbonyl group, aldehyde group, carbonyl group, nitro group, substituted or unsubstituted C1-C30 alkyl group, substituted or unsubstituted C1-20 alkoxy group, substituted or unsubstituted C1-20 alkylamino group, substituted or unsubstituted C3-C60 cycloalkyl group, substituted or unsubstituted C1-C60 alkylthio group, substituted or unsubstituted C6-C60 aryl group, substituted or unsubstituted C3-C60 heteroaryl group;

[0011] Z - is a negative ion;

[0012] n is an integer of 1-2;

[0013] wherein the substituents in the substituted C1-C30 alkyl group, substituted C1-20 alkoxy group, substituted C1-20 alkylamino group, substituted C3-C60 cycloalkyl group, substituted C1-C60 alkylthio group, substituted C6-C60 aryl group, substituted C3-C60 heteroaryl group are selected from deuterium, halogen, cyano group, hydroxyl group, sulfonic acid group, arsinic acid group, boronic acid group, carboxyl group, amino group, mercapto group, C1-C60 alkyl group, C2-C60 alkenyl group, C3-C60 cycloalkyl group, C2-C60 alkynyl group, C3-C60 cycloalkyl group, C1-C60 heterocycloalkyl group, C3-C60 cycloalkenyl group, C1-C60 heterocycloalkenyl group, C6-C60 aryl group.

[0014] In the present application, the term "substituent" has the usual meaning known in the art, referring to a chemical moiety covalently attached to or, where appropriate, fused to a parent core group.

[0015] In the present application, the term "substituted or unsubstituted" means that the functional group recited after the term can or can not have a substituent (hereinafter, the substituents will be collectively referred to as Rcfor the sake of convenience of description). For example, "substituted or unsubstituted aryl group" means an aryl group having a substituent Rc, or an unsubstituted aryl group. The substituents Rcmentioned above, for example, can be deuterium, halogen, cyano group, C1-C60 alkyl group, C3-C60 cycloalkyl group, C6-C60 aryl group, C1-C60 heteroaryl group, and optionally, for example, deuterium, halogen group, cyano group, alkyl group, haloalkyl group, trialkylsilyl group, deuterated alkyl group, aryl group, heteroaryl group, and the like. Of course, the number of substituents Rcmay be one or more. When two substituents Rcare attached to the same atom, the two substituents Rcmay exist independently or be connected to each other to form a ring with the atom; when two substituents Rcexist adjacently on a functional group, the adjacent substituents Rcmay exist independently or be fused to the functional group to form a ring.

[0016] In the present application, the term "alkyl" whether used alone or as part of another term, means a saturated hydrocarbon group, which can be straight-chain or branched. The term "C1-C60 alkyl" is derived from a monovalent substituent of a straight-chain or branched saturated hydrocarbon having from 1 to 60 carbon atoms, preferably from 1 to 40 carbon atoms, and more preferably from 1 to 20 carbon atoms, and examples thereof include, but are not limited to, methyl, ethyl, propyl, isobutyl, sec-butyl, t-butyl, pentyl, isopentyl, and hexyl.

[0017] In the present application, the term "aryl" and "arylene" includes monocyclic, polycyclic or fused ring aryl groups, which can be interrupted by short non-aromatic units between the rings, and can contain a spiro structure, and aryl includes, but is not limited to, phenyl, biphenyl, terphenyl, naphthyl, phenanthryl, anthryl, fluorenyl, spirobifluorenyl, etc., and arylene includes, but is not limited to, phenylene, biphenylene, terphenylene, naphthylene, phenanthrylene, anthrylene, fluorenylene, spirobifluorenylene, etc.

[0018] As used in the present application, the term "substituted" means that a hydrogen atom in the compound is replaced by another substituent. The position is not limited to a specific position, as long as the hydrogen at that position can be replaced by the substituent. When two or more substituents are present, the two or more substituents can be the same or different.

[0019] In the present application, the term "halogen" means an atom selected from fluorine, chlorine, bromine, and iodine.

[0020] In the present application, unless otherwise specified, a hydrogen atom includes protium, deuterium, and tritium.

[0021] In the present application, the range of the number of carbon atoms is defined in the definition of the group, and the number of carbon atoms thereof is any integer within the defined range, for example, C6-C60 aryl, which represents the number of carbon atoms of the aryl group, can be any integer within the range of 6-60, such as 6, 8, 10, 13, 15, 17, 20, 22, 25, or 30, etc.

[0022] In the present application, unless a group is defined as substituted or unsubstituted, it means unsubstituted.

[0023] It can be understood that, for the present application and the like, which contain the following structure or the like:

[0024]

[0025] Preferably, the fluorescent dye compound has the structure shown below:

[0026]

[0027] wherein R1-R3, Z -, n is as defined above.

[0028] Preferably, Z － selected from Cl - , Br - , I - , OTs － , CIO4 - , PF6 - ;

[0029] Preferably, R1, R2, R3 are selected from any one of hydrogen, halogen, benzyl, ester group, amide group, carbonyl, aldehyde group, carbonyl, nitro, substituted or unsubstituted C1-C15 alkyl, substituted or unsubstituted C1-10 alkoxy, substituted or unsubstituted C1-10 alkylamino, substituted or unsubstituted C3-C30 cycloalkyl, substituted or unsubstituted C1-C30 alkylthio, substituted or unsubstituted C6-C30 aryl, substituted or unsubstituted C3-C30 heteroaryl;

[0030] Preferably, R1, R2, R3 are selected from any one of hydrogen, halogen, benzyl, ester group, amide group, carbonyl, aldehyde group, carbonyl, nitro, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-5 alkoxy, substituted or unsubstituted C1-5 alkylamino, substituted or unsubstituted C3-C15 cycloalkyl, substituted or unsubstituted C1-C150 alkylthio, substituted or unsubstituted C6-C15 aryl, substituted or unsubstituted C3-C15 heteroaryl;

[0031] Preferably, R1, R2, R3 are selected from any one of hydrogen, halogen, benzyl, ester group, amide group, carbonyl, aldehyde group, carbonyl, nitro, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-5 alkoxy, substituted or unsubstituted C1-5 alkylamino, substituted or unsubstituted C3-C15 cycloalkyl, substituted or unsubstituted C1-C150 alkylthio, substituted or unsubstituted C6-C15 aryl, substituted or unsubstituted C3-C15 heteroaryl;

[0032] wherein the substituents of the substituted C1-C6 alkyl, substituted C1-5 alkoxy, substituted C1-5 alkylamino, substituted C3-C15 cycloalkyl, substituted C1-C15 alkylthio, substituted C6-C15 aryl, substituted C3-C15 heteroaryl are selected from the group consisting of deuterium, halogen, cyano, hydroxyl, sulfonic acid group, arsinic acid group, boronic acid group, carboxyl, amino, mercapto, C1-C60 alkyl, C2-C60 alkenyl, C3-C60 cycloalkyl, C2-C60 alkynyl, C3-C60 cycloalkyl, C1-C60 heterocycloalkyl, C3-C60 cycloalkenyl, C1-C60 heterocycloalkenyl, C6-C60 aryl;

[0033] Preferably, R1 is selected from the group consisting of H, D, F, Cl, Br, I, benzyl, ester, amide, carbonyl, aldehyde, carbonyl, nitro, substituted or unsubstituted A group, wherein the A group is selected from the group consisting of phenyl, naphthyl, biphenyl, terphenyl, phenanthryl, fluoranthene, triphenylene, dimethylfluorenyl, diphenylfluorenyl, spirobifluorenyl, benzo dimethylfluorenyl, benzo diphenylfluorenyl, benzo spirobifluorenyl, benzofuranyl, dibenzofuranyl, benzothiophenyl, dibenzothiophenyl, carbazolyl, benzo carbazolyl, dibenzo carbazolyl;

[0034] Preferably, R1 is selected from the group consisting of H, D, F, Cl, Br, I, benzyl, ester, amide, carbonyl, aldehyde, carbonyl, nitro, substituted or unsubstituted A group, wherein the A group is selected from the group consisting of phenyl, naphthyl, biphenyl, terphenyl, phenanthryl, fluoranthene, triphenylene, dimethylfluorenyl, diphenylfluorenyl, spirobifluorenyl, benzo dimethylfluorenyl, benzo diphenylfluorenyl, benzo spirobifluorenyl, benzofuranyl, dibenzofuranyl, benzothiophenyl, dibenzothiophenyl, carbazolyl, benzo carbazolyl, dibenzo carbazolyl;

[0035] Preferably, R2 and R3 are each independently selected from the group consisting of hydrogen, hydroxyl, substituted or unsubstituted B group, wherein the B group is selected from the group consisting of methyl, phenyl, naphthyl, biphenyl, terphenyl, phenanthryl, fluoranthene, triphenylene, dimethylfluorenyl, diphenylfluorenyl, spirobifluorenyl, benzo dimethylfluorenyl, benzo diphenylfluorenyl, benzo spirobifluorenyl, benzofuranyl, dibenzofuranyl, benzothiophenyl, dibenzothiophenyl, carbazolyl, benzo carbazolyl, dibenzo carbazolyl, or a group having the following structure:

[0036]

[0037] Preferably, R2 and R3 are each independently selected from the group consisting of hydrogen, hydroxyl, substituted or unsubstituted B group, wherein the B group is selected from the group consisting of methyl, phenyl, naphthyl, biphenyl, terphenyl, phenanthryl, fluoranthene, triphenylene, dimethylfluorenyl, diphenylfluorenyl, spirobifluorenyl, benzo dimethylfluorenyl, benzo diphenylfluorenyl, benzo spirobifluorenyl, benzofuranyl, dibenzofuranyl, benzothiophenyl, dibenzothiophenyl, carbazolyl, benzo carbazolyl, dibenzo carbazolyl, or a group having the following structure:

[0038] Preferably, R is selected from one of the following: -C-N-S1, -C=C-S2;

[0039] wherein S1is selected from hydrogen, substituted or unsubstituted C1-C8alkyl, substituted or unsubstituted C3-C6cycloalkyl, substituted or unsubstituted C6-C12aryl, substituted or unsubstituted C5-C12heteroaryl, substituted or unsubstituted amido;

[0040] wherein each of the substituents in the substituted C1-C8alkyl, substituted C3-C6cycloalkyl, substituted C6-C12aryl, substituted C5-C12heteroaryl, substituted amido is independently selected from halogen, nitro, cyano, hydroxyl, sulfonic, arsine, boronic, carboxyl, amino, mercapto, C1-C6alkyl, C2-C6alkenyl, C3-C6cycloalkyl, C3-C6heterocycloalkyl, C2-C6alkynyl, C3-C10cycloalkenyl, C6-C10aryl, C3-C10heteroaryl, amido;

[0041] each S2is independently selected from substituted or unsubstituted onium, wherein each of the substituents in the substituted group is independently selected from substituted or unsubstituted C1-C8alkyl, substituted or unsubstituted C3-C6cycloalkyl, substituted or unsubstituted C6-C12aryl, substituted or unsubstituted C5-C12heteroaryl, substituted or unsubstituted amido;

[0042] wherein each of the substituents in the substituted C1-C8alkyl, substituted C3-C6cycloalkyl, substituted C6-C12aryl, substituted C5-C12heteroaryl, substituted amido is independently selected from halogen, cyano, hydroxyl, sulfonic, arsine, boronic, carboxyl, amino, mercapto, C1-C6alkyl, C2-C6alkenyl, C3-C6cycloalkyl, C3-C6heterocycloalkyl, C2-C6alkynyl, C3-C10cycloalkenyl, C6-C10aryl, C3-C10heteroaryl;

[0043] Preferably, S1is selected from substituted or unsubstituted D groups, wherein the D groups include: methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, cyclopropanyl, cyclopentanoyl, cyclohexanoyl, phenyl, tolyl, ethylphenyl, naphthyl, pyridyl, quinolyl, amido;

[0044] wherein each of the substituents in the substituted D groups is selected from halogen, nitro, cyano, hydroxyl, sulfonic, arsine, boronic, carboxyl, amino, mercapto, C1-C6alkyl, C2-C6alkenyl, C3-C6cycloalkyl, C3-C6heterocycloalkyl, C2-C6alkynyl, C3-C10cycloalkenyl, C6-C10aryl, C3-C10heteroaryl;

[0045] Preferably, the substituents in the substituted D group are selected from the group consisting of halogen, nitro, cyano, hydroxy, sulfonic acid, arsine acid, boronic acid, carboxylic acid, amino, mercapto, methyl, dimethylamino, tetrahydrothiophene, tetrahydrofuran, phalloidin, taxol, amide, or a group having the following structure:

[0046]

[0047] Preferably, each S2 is independently selected from the group consisting of substituted or unsubstituted pyridinium, substituted or unsubstituted quinolinium, substituted or unsubstituted indolium, substituted or unsubstituted benzindolium, substituted or unsubstituted benzothiazolium;

[0048] wherein each substituent in the substituted pyridinium, substituted quinolinium, substituted indolium, substituted benzindolium, substituted benzothiazolium is independently selected from the group consisting of halogen, cyano, hydroxy, sulfonic acid, arsine acid, boronic acid, carboxylic acid, amino, mercapto, C1-C6 alkyl, C2-C6 alkenyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, C2-C6 alkynyl, C3-C10 cycloalkenyl, C6-C10 aryl, C3-C10 heteroaryl;

[0049] Further preferably, each substituent in the substituted pyridinium, substituted quinolinium, substituted indolium, substituted benzindolium, substituted benzothiazolium is independently selected from the group consisting of halogen, cyano, hydroxy, sulfonic acid, arsine acid, boronic acid, carboxylic acid, amino, mercapto, or a group having the following structure:

[0050]

[0051] Preferably, S1 is selected from one of the following groups:

[0052]

[0053] wherein X is halogen;

[0054] In this application, phalloidin is a phalloidin group; taxol is a taxol group;

[0055] Preferably, S2 is selected from one of the following groups:

[0056]

[0057] Preferably, the fluorescent dye compound has one of the following structures:

[0058] Preferably, the structure of the fluorescent dye compound comprises the following structure:

[0059]

[0060]

[0061]

[0062]

[0063]

[0064]

[0065] wherein R1-R3, Z - , n are defined as above.

[0066] Preferably, R1, R2, R3 are selected from any one of hydrogen, halogen, benzyl, ester group, amide group, carbonyl group, aldehyde group, carbonyl group, nitro group, substituted or unsubstituted C1-C15 alkyl, substituted or unsubstituted C1-10 alkoxy, substituted or unsubstituted C1-10 alkylamino, substituted or unsubstituted C3-C30 cycloalkyl, substituted or unsubstituted C1-C30 alkylthio, substituted or unsubstituted C6-C30 aryl, substituted or unsubstituted C3-C30 heteroaryl;

[0067] wherein the substituents in substituted C1-C15 alkyl, substituted C1-10 alkoxy, substituted C1-10 alkylamino, substituted C3-C30 cycloalkyl, substituted C1-C30 alkylthio, substituted C6-C30 aryl, substituted C3-C30 heteroaryl are selected from deuterium, halogen, cyano, hydroxyl, sulfonic acid group, arsinic acid group, boronic acid group, carboxyl, amino, mercapto, C1-C60 alkyl, C2-C60 alkenyl, C3-C60 cycloalkyl, C2-C60 alkynyl, C3-C60 cycloalkyl, C1-C60 heterocycloalkyl, C3-C60 cycloalkenyl, C1-C60 heterocycloalkenyl, C6-C60 aryl;

[0068] Preferably, R1, R2, R3 are selected from any one of hydrogen, halogen, benzyl, ester group, amide group, carbonyl group, aldehyde group, carbonyl group, nitro group, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-5 alkoxy, substituted or unsubstituted C1-5 alkylamino, substituted or unsubstituted C3-C15 cycloalkyl, substituted or unsubstituted C1-C150 alkylthio, substituted or unsubstituted C6-C15 aryl, substituted or unsubstituted C3-C15 heteroaryl; and / or,

[0069] R2, R3are each independently selected from any one of hydrogen-substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-5 alkoxy, substituted or unsubstituted C1-5 alkylamino, substituted or unsubstituted C3-C15 cycloalkyl, substituted or unsubstituted C1-C150 alkylthio, substituted or unsubstituted C6-C15 aryl, substituted or unsubstituted C3-C15 heteroaryl;

[0070] wherein the substituents of the substituted C1-C6 alkyl, substituted C1-5 alkoxy, substituted C1-5 alkylamino, substituted C3-C15 cycloalkyl, substituted C1-C150 alkylthio, substituted C6-C15 aryl, substituted C3-C15 heteroaryl are selected from deuterium, halogen, cyano, hydroxyl, sulfonic acid group, arsinic acid group, boronic acid group, carboxyl, amino, mercapto, C1-C60 alkyl, C2-C60 alkenyl, C3-C60 cycloalkyl, C2-C60 alkynyl, C3-C60 cycloalkyl, C1-C60 heterocycloalkyl, C3-C60 cycloalkenyl, C1-C60 heterocycloalkenyl, C6-C60 aryl;

[0071] Preferably, R1is selected from H, D, F, Cl, Br, I, benzyl, ester, amide, carbonyl, aldehyde, carbonyl, nitro, substituted or unsubstituted A group, wherein the A group is selected from methyl, methoxy, methoxyphenyl, phenyl, naphthyl, biphenyl, terphenyl, phenanthryl, fluoranthryl, triphenylenyl, dimethylfluorenyl, diphenylfluorenyl, spirobifluorenyl, benzo dimethylfluorenyl, benzo diphenylfluorenyl, benzo spirobifluorenyl, benzofuranyl, dibenzofuranyl, benzothiophenyl, dibenzothiophenyl, carbazolyl, benzo carbazolyl, dibenzo carbazolyl;

[0072] wherein the substituents of the substituted A group are selected from deuterium, halogen, cyano, hydroxyl, sulfonic acid group, arsinic acid group, boronic acid group, carboxyl, amino, mercapto, C1-C50 alkyl, C2-C5 alkenyl, C3-C10 cycloalkyl, C2-C10 alkynyl, C3-C5 cycloalkyl, C1-C10 heterocycloalkyl, C3-C10 cycloalkenyl, C1-C10 heterocycloalkenyl, C6-C10 aryl;

[0073] Preferably, R2, R3are each independently selected from hydrogen, hydroxyl, substituted or unsubstituted B group, wherein the B group is selected from methyl, phenyl, naphthyl, biphenyl, terphenyl, phenanthryl, fluoranthryl, triphenylenyl, dimethylfluorenyl, diphenylfluorenyl, spirobifluorenyl, benzo dimethylfluorenyl, benzo diphenylfluorenyl, benzo spirobifluorenyl, benzofuranyl, dibenzofuranyl, benzothiophenyl, dibenzothiophenyl, carbazolyl, benzo carbazolyl, dibenzo carbazolyl, or a group having the following structure:

[0074]

[0075] wherein, the substituent in the substituted B group is deuterium, halogen, cyano, hydroxyl, sulfonic acid group, arsinic acid group, boric acid group, carboxyl, amino, mercapto, C1-C50 alkyl, C2-C5 alkenyl, C3-C10 cycloalkyl, C2-C10 alkynyl, C3-C5 cycloalkyl, C1-C10 heterocycloalkyl, C3-C10 cycloalkenyl, C1-C10 heterocycloalkenyl, C6-C10 aryl.

[0076] The application further provides a preparation method of the fluorescent dye compound, comprising the following steps:

[0077] performing an aldehyde group modification reaction on a compound with the structure shown in the following formula A to obtain the fluorescent dye compound;

[0078] Cy-CHO formula A;

[0079] wherein Cy- is selected from one of the following structures:

[0080]

[0081] wherein, R1-R3, Z - , n are defined as above;

[0082] Preferably, the aldehyde group modification reaction is selected from an aldehyde group-amino Schiff base reaction and an aldehyde group-methyl dehydration condensation reaction.

[0083] Preferably, the preparation method of the compound with the structure shown in formula A comprises the following steps:

[0084] adding sodium borohydride into a solution of the Cy compound, then performing evaporation of solvent, washing and drying to obtain a first reaction product, then adding the first reaction product into a solution of phosphorus oxychloride to react, then performing extraction to obtain a second reaction product, reacting the second product with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone, and then purifying to obtain Cy-CHO;

[0085] wherein, the Cy compound is selected from one of the compounds with the following structure:

[0086]

[0087] Preferably, the solvent in the solution of the Cy compound is selected from one or more of acetonitrile, ethyl acetate, methanol, ethanol, dichloromethane, chloroform, toluene and DMF;

[0088] Preferably, the molar ratio of the Cy compound to sodium borohydride is (2-4):1;

[0089] Preferably, the temperature for the reaction of adding sodium borohydride into the solution of the Cy compound is 20-40°C, and the reaction time is 5-10 min.

[0090] Preferably, the solvent in the phosphorus oxychloride solution is selected from DMF;

[0091] Preferably, the molar ratio of the first reaction product to the phosphorus oxychloride is 1:(30-50);

[0092] Preferably, the temperature for the reaction of the first reaction product with the phosphorus oxychloride solution is 60-80℃; and the reaction time is 4-6h;

[0093] Preferably, the molar ratio of the second product to 2,3-dichloro-5,6-dicyano-1,4-benzoquinone is (4-6):1;

[0094] Preferably, the temperature for the reaction of the second product with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone is 20-40℃; and the reaction time is 10-30min.

[0095] The application also provides the application of the fluorescent dye compound prepared by the above-mentioned method in live cell fluorescence imaging and super-resolution imaging.

[0096] The application also provides a preparation method of the Cy compound, comprising the following steps:

[0097] E01: heating and reacting a compound of formula VII with a nucleophile 2-bromoethyl-1,3-dioxolane to obtain a compound of formula V;

[0098] E02: heating and reacting a compound of formula VIII with a nucleophile 2-bromoethyl-1,3-dioxolane to obtain a compound of formula VI;

[0099] E03: preparing a compound of formula IV by reacting the compound of formula V or VI in acetic anhydride under the catalysis of sodium acetate;

[0100]

[0101] E04: dissolving the compound (n=1) of formula IV in chloroform, and performing a condensation ring-closing reaction under the condition of concentrated sulfuric acid to obtain a compound of formula IX;

[0102]

[0103] Optionally, when R1 is hydrogen, the reaction process of step E04 is as follows:

[0104]

[0105] Optionally, when R1 is methyl, the reaction process of step E04 is as follows:

[0106]

[0107] Optionally, when R1 is methoxy, the reaction process of step E04 is as follows:

[0108]

[0109] Optionally, when R1 is phenyl, the reaction process of step E04 is as follows:

[0110]

[0111] Optionally, when R1 is substituted phenyl, the reaction process of step E04 is as follows:

[0112]

[0113] wherein X is selected from F, Cl, Br, I or -OCH3.

[0114] The present application also provides a method for preparing other Cy compounds, comprising the following steps:

[0115]

[0116] S1 reacting a compound of formula VII with a compound having the structure of R3-Ar-Z at 80-120℃ for 6-36h to obtain a compound having the structure of formula V'; wherein the reaction solvent is selected from toluene or acetonitrile; the molar ratio of the compound of formula VII to the compound having the structure of R3-Ar-Z is 1:1-1:4;

[0117] S2 reacting a compound of formula VIII with a compound having the structure of R2-Ar-Z at 80-120℃ for 6-36h to obtain a compound having the structure of formula VI'; wherein the reaction solvent is selected from toluene or acetonitrile; the molar ratio of the compound of formula VII to the compound having the structure of R2-Ar-Z is 1:1-1:4;

[0118] S3 reacting the compound having the structure of formula V' with the compound having the structure of formula VI' to obtain a compound having the structure of formula IV';

[0119] wherein R1, R2, R3, Z are the same as defined above; Ar is selected from C1-C6 alkylene, C6-C15 arylene;

[0120] Preferably, Ar is selected from ethylmethyl, methylene, phenylene, naphthylene;

[0121] when Z is Cl - , Br - , the reaction in steps S1 and S2 is carried out in the presence of KI or KBr as catalyst.

[0122] Advantages of the present application:

[0123] The fluorescent dye compound provided by the present application has a Cy-R structure, wherein R is a group obtained by reacting with an aldehyde group; by introducing the R group on the basis of the Cy structure, the photophysical properties of the compound are greatly improved compared to the compound without R, thereby reducing the saturation loss laser power of the dye, further reducing the saturation loss power in stimulated emission loss super-resolution imaging, enhancing the permeability and compatibility of the compound for living cells, and enabling better use of the compound for stimulated emission loss super-resolution imaging of living cells, thereby improving the biological imaging application range of the dye; and the compound with Cy-R has good light bleaching resistance and light stability, thereby enabling the compound to be applied to super-resolution imaging of living cells to characterize more internal details of the cells.

[0124] Further, the compound provided by the present application can provide nanoscale imaging of living cells, provide more detailed structural information that cannot be obtained by conventional microscopic imaging, enable longer and clearer observation of nanoscale microstructures and dynamic changes, and have important significance for in-depth understanding of life phenomena and research on related diseases. BRIEF DESCRIPTION OF DRAWINGS

[0125] In order to more clearly illustrate the specific embodiments of the present application or the technical solutions in the prior art, the drawings needed in the specific embodiments or prior art description will be briefly introduced below. Obviously, the drawings in the following description are some embodiments of the present application, and those skilled in the art can also obtain other drawings according to these drawings without creative labor.

[0126] Figure 1 The STED super-resolution microscopic imaging diagram of the local mitochondria of COS-7 cells corresponding to the compound Cy3B-1 in this embodiment 4;

[0127] Figure 2 The STED super-resolution microscopic imaging diagram of the local mitochondria of COS-7 cells corresponding to the fluorescent dye compound Ia6-1 in this embodiment 4;

[0128] Figure 3 The STED super-resolution microscopic imaging diagram of the local mitochondria of COS-7 cells corresponding to the fluorescent dye compound Ia9-1 in this embodiment 5.

[0129] Figure 4 The STED super-resolution microscopic imaging diagram of the local microfilament microtubule of Hala cells corresponding to the fluorescent dye compound Ia29-1 in this embodiment 7.

[0130] Figure 5COS-7 cell local mitochondria STED super-resolution microscopy imaging map corresponding to the fluorescent dye compound Ia37-1 in this Example 8.

[0131] Figure 6 COS-7 cell mitochondria STED super-resolution microscopy imaging map corresponding to the fluorescent dye compound IIa5-1 in this Example 11.

[0132] Figure 7 COS-7 cell local mitochondria STED super-resolution microscopy imaging map corresponding to the fluorescent dye compound IIa46-1 in this Example 13.

[0133] Figure 8 COS-7 cell local mitochondria STED super-resolution microscopy imaging map corresponding to the fluorescent dye compound IIa53-1 in this Example 14. DETAILED DESCRIPTION

[0134] When specific experimental procedures or conditions are not mentioned in the examples, the operation or conditions according to the conventional experimental procedures described in the literature in the art can be used. When the used reagents or instruments are not mentioned the manufacturer, they are all the conventional reagent products that can be obtained by purchase.

[0135] Those skilled in the art will recognize that the chemical reactions described in this application can be used to prepare many of the heterocyclic compounds of this application, and that other methods for preparing the compounds of this application are also within the scope of this application. For example, the synthesis of those non-exemplified compounds according to this application can be successfully performed by modifications apparent to those skilled in the art, by analogy with the above described methods, by application of known chemical reactions, or by application of known chemical reagents other than those described above, or by application of known chemical reagents under conditions other than those described above. The compounds of this application for which no synthetic methods are described in the application are obtained by commercial sources.

[0136] Synthesis of some intermediates:

[0137] 1. Synthesis of compound with the following structure:

[0138]

[0139] Synthesis of compound VI-1, comprising the following steps:

[0140] To a suspension of KI (3.2 g, 20 mol) in distilled MeCN (50 mL), under nitrogen atmosphere, 2-bromoethyl-l,3-dioxolane (5.2 mL, 44 mmol) was added, and the reaction was stirred at 100 °C for 1 h in the dark. 2,3,3-Trimethyl-3H-indole (6.4 mL, 40 mmol) was added, and the reaction was stirred at reflux for 48 h. The reaction was allowed to cool to room temperature and filtered to remove the KBr salt. The solvent was removed under vacuum, and the product was purified by column chromatography (purification in MeOH / DCM and 0.5% Et3N) to give compound VI-1 as a dark red oil. Compound VI-1 structural characterization data are as follows: 1 H-NMR (400 MHz, DMSO-d6): 7.34 (t, 1H), 7.06 (t, 1H), 6.72 (t, 1H), 6.36 (t, 1H), 5.46 (s, 1H), 4.40 (d, 2H), 4.00 (m, 1H), 3.97 (d, 1H), 3.87 (t, 2H), 3.77 (t, 2H), 1.82 (d, 2H), 1.79 (m, 6H). 13 C-NMR (100 MHz, DMSO-d6): 26.4, 29.1, 37.2, 57.2, 64.7, 73.4, 106.9, 108.6, 16.8, 120.8, 126.7, 140.9, 146.6, 160.4. HRMS-ESI: m / z calcd for C + for C 16 H 22 NO2 + , 259.1572; found, 259.1576.

[0141] The synthesis of compound VI-2 was similar to the analogous, with the exception that 5-methyl-2,3,3-trimethyl-3H-indole was added instead of 2,3,3-trimethyl-3H-indole. Compound VI-2 structural characterization data are as follows: HRMS-ESI: m / z calcd for C + for C 16 H 22 NO2 + , 274.3835; found, 274.3830.

[0142] The synthesis of compound VI-3 was similar to the analogous, with the exception that 5-methoxy-2,3,3-trimethyl-3H-indole was added instead of 2,3,3-trimethyl-3H-indole. Compound VI-3 structural characterization data are as follows: HRMS-ESI: m / z calcd for C + for C 17 H 24 NO3 +, 290.3825, found, 290.3829.

[0143] The synthesis of compound VI-4 is similar to the above, except that 2,3,3- trimethyl-5-phenyl-3H-indole is added instead of 2,3,3-trimethyl-3H-indole. The structural characterization data of compound VI-4 are as follows: HRMS-ESI: m / z calcd M + for C 22 H 26 NO2 + , 336.4545, found, 336.4550.

[0144] When X is F in formula VI-5: the synthesis of compound VI-5-1 is similar to the above, except that 5-fluorophenyl-2,3,3-trimethyl-3H-indole is added instead of 2,3,3-trimethyl-3H-indole. The structural characterization data of compound VI-5-1 are as follows: HRMS-ESI: m / z calcd M + for C 22 H 25 FNO2 + , 354.4449, found, 354.4450.

[0145] When X is Cl in formula VI-5: the synthesis of compound VI-5-2 is similar to the above, except that 5-chlorophenyl-2,3,3-trimethyl-3H-indole is added instead of 2,3,3-trimethyl-3H-indole. The structural characterization data of compound VI-5-2 are as follows: HRMS-ESI: m / z calcd M + for C 22 H 25 ClNO2 + , 370.8965, found, 370.8962.

[0146] When X is Br in formula VI-5: the synthesis of compound VI-5-3 is similar to the above, except that 5-bromophenyl-2,3,3-trimethyl-3H-indole is added instead of 2,3,3-trimethyl-3H-indole. The structural characterization data of compound VI-5-3 are as follows: HRMS-ESI: m / z calcd M + for C 22 H 25 BrNO2 + , 415.3505, found, 415.3510.

[0147] When X is methoxyphenyl in Formula VI-5: the synthesis of compound VI-5-5 is similar to the above, except that 5-methoxyphenyl-2,3,3-trimethyl-3H-indolium is used to replace 2,3,3-trimethyl-3H-indolium. The structure characterization data of compound VI-5-5 is as follows: HRMS-ESI: m / z calcd for C + for C 22 H 26 NO2 + ,336.4545, found, 336.4550.

[0148] When X is methoxyphenyl in Formula VI-5: the synthesis of compound VI-5-5 is similar to the above, except that 5-methoxyphenyl-2,3,3-trimethyl-3H-indolium is used to replace 2,3,3-trimethyl-3H-indolium. The structure characterization data of compound VI-5-5 is as follows: HRMS-ESI: m / z calcd for C + for C 23 H 28 NO3 + ,366.4805, found, 366.4809.

[0149] The synthesis of Cy3-type compounds is shown in the following reaction formula:

[0150]

[0151] The synthesis of compound IV-3-1 (wherein Y is H) comprises the following steps: stirring 1-ethyldioxolane-2,3,3-trimethylindolium (compound VI-1 (Y is H)) (5.18 g, 20 mmol), N,N'-diphenylformamidine (1.96 g, 10 mmol) and sodium acetate (0.41 g, 5 mmol) in 5-8 mL acetic anhydride under nitrogen atmosphere and dark conditions at 80°C for 15 h, washing with DCM and saturated brine, drying with anhydrous sodium sulfate, removing the solvent in vacuum, and purifying by column chromatography (purified in MeOH / DCM = 1:20) to obtain a dark green metallic luster solid. The structure characterization data of compound IV-3-1 (n = 1) is as follows: 1H-NMR (400 MHz, DMSO-d6): 8.95 (d, 1H), 8.12 (d, 2H), 7.44 (d, 1H), 7.34 (t, 1H), 7.30 (d, 1H), 7.06 (t, 1H), 6.72 (t, 1H), 6.55 (d, 1H), 6.36 (t, 1H), 5.64 (t, 1H), 5.46 (m, 2H), 5.33 (d, 1H), 4.01 (d, 2H), 4.0 (s, 4H), 3.87 (m, 4H), 3.77 (t, 2H), 1.82 (d, 1H), 1.79 (m, 6H), 1.6 (s, 2H), 1.44 (m, 6H). 13 C-NMR (100 MHz, DMSO-d6): 173.1, 172.8, 149.5, 146.6, 141.4, 141.2, 140.9, 128.3, 126.7, 125.0, 120.8, 120.1, 116.8, 111.0, 108.6, 107.3, 106.9, 104.8, 104.5, 64.7, 55.0, 51.1, 43.1, 37.7, 31.7, 29.1, 26.8, 24.7. HRMS-ESI: m / z calcd M + for C 33 H 41 N2O4 + , 529.3061; found, 529.3066.

[0152] Compound IV-3-2 (Y is CH3) was synthesized by the following steps: stirring compound VI-2 (Y is CH3) (2.74 g, 10 mmol) and compound V (2.59 g, 10 mmol), N,N'-diphenylformamidine (1.96 g, 10 mmol) and sodium acetate (0.41 g, 5 mmol) in 5-8 mL acetic anhydride under nitrogen atmosphere and dark condition, 80 °C for 15 h, washing with DCM and saturated brine, drying with anhydrous sodium sulfate, removing solvent under vacuum, and purifying by column chromatography (purifying in MeOH / DCM = 1:20) to give dark green metallic luster solid compound IV-3-2 (n = 1), HRMS-ESI: m / z calcd M + for C 34 H 43 N2O4 + 543.7275; found, 543.7278.

[0153] Synthesis of compound IV-3-3 (Y is OCH3) includes the following steps which are similar to the synthesis of compound IV-3-1 except that compound VI-3 (Y is OCH3) is used in place of compound VI-1 (Y is H) to give compound IV-3-3 (n = 1), HRMS-ESI: m / z calcd M + for C 34 H 43 N2O5 + 559.7265; found, 559.7260.

[0154] Synthesis of compound IV-3-4 (Y is ph) includes the following steps which are similar to the synthesis of compound IV-3-1 except that compound VI-4 (Y is ph) is used in place of compound VI-1 (Y is H) to give compound IV-3-4 (n = 1), HRMS-ESI: m / z calcd M + for C 39 H 45 N2O4 + 605.7985; found, 605.7983.

[0155] Synthesis of compound IV-3-5 (Y is phOCH3) includes the following steps which are similar to the synthesis of compound IV-3-1 except that compound VI-5 (Y is phOCH3) is used in place of compound VI-1 (Y is H) to give compound IV-3-5 (n = 1), HRMS-ESI: m / z calcd M + for C 40 H 47 N2O5 + 635.8245; found, 635.8249.

[0156] Synthesis of Cy5-type compounds, as shown in the following reaction scheme:

[0157]

[0158] The synthesis method is similar to that of Cy3-type compounds, except that N, N'-diphenylformamidine is replaced by hydrochloric acid-N-(3-phenylamino-2-propenylidene) aniline to give the corresponding Cy5-type compound.

[0159] The synthesis of compound IV-5-1 (where Y is H) includes the following steps: stirring 1-ethyldioxolane-2,3,3-trimethylindole (compound VI-1 (Y is H)) (5.18 g, 20 mmol), hydrochloric acid-N-(3-phenylamino-2-propenylidene)aniline (2.58 g, 10 mmol) and sodium acetate (0.41 g, 5 mmol) in 5-8 mL of acetic anhydride under nitrogen atmosphere and dark condition at 80 °C for 15 h, washing with DCM and saturated brine, drying with anhydrous sodium sulfate, removing the solvent under vacuum, and purifying by column chromatography (purifying in MeOH / DCM = 1:20) to obtain dark green metallic luster solid. Compound IV-5-1 was obtained, and the structural characterization data are as follows: HRMS-ESI: m / z calcd for C + for C 35 H 43 N2O4 + ,555.7385; found, 555.7381.

[0160] The synthesis of compound IV-5-2 (Y is CH3) includes the following steps: stirring compound VI-2 (Y is CH3) (2.74 g, 10 mmol) and compound V (2.59 g, 10 mmol), hydrochloric acid-N-(3-phenylamino-2-propenylidene)aniline (2.58 g, 10 mmol) and sodium acetate (0.41 g, 5 mmol) in 5-8 mL of acetic anhydride under nitrogen atmosphere and dark condition at 80 °C for 15 h, washing with DCM and saturated brine, drying with anhydrous sodium sulfate, removing the solvent under vacuum, and purifying by column chromatography (purifying in MeOH / DCM = 1:20) to obtain dark green metallic luster solid compound IV-5-2, HRMS-ESI: m / z calcd for C + for C 34 H 43 N2O4 + 569.7655; found, 569.7659.

[0161] The synthesis of compound IV-5-3 (Y is OCH3) includes the following steps which are similar to the synthesis of compound IV-5-1 except that compound VI-3 (Y is OCH3) is used instead of compound VI-1 (Y is H) to obtain compound IV-5-3 (n = 2), HRMS-ESI: m / z calcd for C + for C 36 H 45 N2O5 + 585.7645; found, 585.7648.

[0162] Synthesis of compound IV-5-4 (Y = ph) includes the following steps similar to the synthesis of compound IV-5-1 except that compound VI-4 (Y = ph) is used in place of compound VI-1 (Y = H) to give compound IV-5-4 (n = 2). HRMS-ESI: m / z calcd for C + for C 41 H 47 N2O4 + 631.8365; found, 631.8361.

[0163] Synthesis of compound IV-5-5 (Y = phOCH3) includes the following steps similar to the synthesis of compound IV-5-1 except that compound VI-5 (Y = phOCH3) is used in place of compound VI-1 (Y = H) to give compound IV-5-5 (n = 2). HRMS-ESI: m / z calcd for C + for C 42 H 49 N2O5 + 661.8625; found, 661.8627.

[0164] Synthesis of compound Cy3B-1 includes the following steps:

[0165]

[0166] Synthesis of compound Cy3B-1 includes the following steps:

[0167]

[0168] Compound IV-3-1 was placed in a conical flask and anhydrous dichloromethane was added to dissolve it completely. 70-80% concentrated sulfuric acid was added dropwise while stirring vigorously. The reaction was allowed to proceed at room temperature for 15-60 min, which was monitored by TLC. A significant change in the solution was observed in terms of fluorescence color and intensity. The dichloromethane / chloroform layer was collected and the solvent was evaporated under vacuum to give the corresponding metallic luster solid, which was compound Cy3B-1. The structural characterization data of compound Cy3B-1 are as follows: 1H-NMR (400 MHz, DMSO-d6): 8.95 (d, 1H), 8.12 (d, 1H), 7.47 (m, 1H), 7.44 (d, 1H), 7.34 (t, 1H), 7.30 (d, 1H), 7.06 (t, 1H), 6.72 (t, 1H), 6.36 (t, 1H), 3.88 (t, 2H), 3.09 (d, 1H), 2.99 (d, 1H), 1.81 (t, 1H), 1.79 (m, 6H), 1.62 (d, 1H), 1.56 (d, 1H), 1.44 (m, 6H), 1.4 (d, 1H), 1.36 (s, 1H), 1.3 (s, 1H). 13 C-NMR (100 MHz, DMSO-d6): 183.5, 164.4, 146.6, 141.2, 141.4, 140.9, 135.2, 128.3, 127.9, 126.7, 125.0, 120.8, 120.1, 116.8, 111.0, 109.6, 108.6, 78.2, 76.8, 52.8, 50.9, 49.0, 43.3, 31.3, 27.1, 26.7, 25.0. HRMS-ESI: m / z calcd M + for C 29 H 31 N2O + , 423.2431, found, 423.2435.

[0169] Example 1

[0170] The present synthesis example provides a method for preparing compound Cy3-CHO-1, comprising the following steps:

[0171]

[0172] The compound Cy3-H and MeOH were mixed, sodium borohydride was added for reduction, and the reaction was carried out at room temperature for 15-30 min until the solution color was obviously light yellow. The solvent was evaporated, washed with DCM / H2O, dried with anhydrous sodium sulfate, and the solvent was evaporated to obtain the first reaction product. Phosphorus oxychloride and DMF were stirred in an ice bath for 30 min, and the first reaction product was gradually added dropwise. The mixture was stirred at room temperature for 25 min, and then the temperature was increased to 70°C for reaction for 5 h. After the reaction was completed, the mixture was poured into 40 mL of ice water, NaHCO3 was added to quench the reaction, and DCM was added for extraction. The mixture was dried with anhydrous sodium sulfate, the solvent was evaporated, DDQ was added for oxidation, and column chromatography was used for purification (purification was carried out in MeOH / DCM=1:20) to obtain a green metallic luster solid, which was compound Cy3-CHO-1. The structure characterization data of compound Cy3-CHO-1 are as follows: HRMS-ESI: m / z calcd M +for C 34 H 41 N2O5 + C 28 H 33 N2O + ,413.5845found,413.5840.

[0173] Other compounds similar to Cy3-CHO-1 (as the following reaction product) are prepared in a similar manner to the preparation of Cy3-CHO-1, except that different starting materials are used:

[0174]

[0175] wherein Y is selected from CH3, OCH3, Ph, PhOCH3, F, Cl, Br or I.

[0176] Example 2

[0177] This synthesis example provides a method for preparing compound Cy5-CHO-1,

[0178]

[0179] The preparation process is similar to synthesis example 1, except that compound Cy3-H is replaced by compound Cy5-H. The structural characterization data of compound Cy5-CHO-1 is as follows: HRMS-ESI: m / z calcd M + for C 36 H 43 N2O5 + C 36 H 38 N2O4S,594.7700found,594.7720.

[0180] Other compounds similar to Cy5-CHO-1 (as the following reaction product) are prepared in a similar manner to the preparation of Cy5-CHO-1, except that different starting materials are used:

[0181]

[0182] wherein Y is selected from CH3, OCH3, Ph, PhOCH3, F, Cl, Br or I.

[0183] Example 3

[0184] This synthesis example provides a method for preparing compound Cy3B-CHO-1, comprising the following steps:

[0185]

[0186] The preparation process is similar to that in Example 1, except that compound Cy3-H is replaced with compound Cy3B-H. The structural characterization data of compound Cy3B-CHO-1 are as follows: 1 H-NMR(400MHz,DMSO-d6):9.88(d,1H),8.95(t,1H),8.12(t,1H),7.85(m,1H),7.74(m,1H),7.44(t,1H),7.30(t,1H), 7.22(m,1H),6.86(m,1H),3.88(m,2H),3.09(d,2H),1.81(t,2H),1.79(m,6H),1.63(d,2H),1.44(m,6H),1.36(d,2H). 13 C-NMR(100MHz,DMSO-d6):183.5,164.4,146.6,141.6,141.2,140.9,135.2,128.3,127.9,126.7,125.0,120.8 ,120.1,116.8,111.0,109.6,108.6,78.2,76.8,52.8,50.9,49.0,43.3,26.7,31.3,27.0,25.0.HRMS-ESI:m / z calcdM + for C 30 H 31 N2O2 + ,451.2380found,451.2386.

[0187] Other substances similar to compound Cy3B-CHO-1 (reaction products shown below) are prepared using methods similar to those for compound Cy3B-CHO-1, the difference being the use of different starting materials:

[0188]

[0189] Wherein, Y is selected from CH3,OCH3,Ph,PhOCH3.

[0190] Example 4

[0191] This embodiment provides compound I a6 Preparation method of -1, compound I a6 The structure of -1 is as follows:

[0192]

[0193] The preparation method of the raw material compound Cy5-CHO-CH3-ph-OH having the following structure is similar to that of Cy5-CHO-1, except that different initial raw materials are used.

[0194]

[0195] Compound I a6 The preparation of -1 specifically comprises the following steps:

[0196] Compound Cy5-CHO-CH3-ph-OH (0.265 g, 0.5 mmol) and butylamine (0.073 g, 1 mmol) were dissolved in MeOH (6 mL), 2 drops of DIPEA were added dropwise, and stirred at 45-55°C under nitrogen atmosphere overnight. After the reaction was completed, KBH4 (0.54 g, 0.5 mmol) was added to the mixture for reduction reaction, the solvent was evaporated, extracted with DCM / H2O, and DDQ (0.036 g, 0.25 mmol) was added to the CH2Cl2 solution for oxidation, and purified by column chromatography (MeOH / DCM) to obtain compound I a6 -1. Compound I a6 The structure of -1 is characterized as follows: 1 H-NMR (400 MHz, DMSO-d6): 8.90 (d, 1H), 7.73 (t, 1H), 7.25 (d, 2H), 7.22 (d, 1H), 7.18 (m, 1H), 7.16 (d, 2H), 6.70 (m, 1H), 6.52 (d, 2H), 6.55 (s, 1H), 5.56 (s, 1H), 5.33 (s, 1H), 4.81 (s, 1H), 3.64 (t, 2H), 3.59 (t, 2H), 3.07 (d, 2H), 2.60 (d, 2H), 2.53 (d, 2H), 2.31 (t, 3H), 1.79 (m, 6H), 1.44 (t, 6H), 1.38 (t, 2H), 0.89 (d, 3H). 13 C-NMR (100 MHz, DMSO-d6): 173.1, 172.8, 148.0, 144.9, 140.7, 138.4, 138.1, 137.4, 134.5, 130.4, 128.0, 125.0, 127.7, 125.9, 126.2, 127.7, 128.6, 125.7, 124.5, 113.0, 104.8, 104.5, 63.4, 58.2, 55.3, 52.9, 51.4, 49.5, 49.2, 32.7, 26.8, 24.7, 21.6, 20.1, 13.8. HRMS-ESI: m / z calcd M + for C 40 H50 N3O + ,558.8595found,558.8591.。

[0197] Example 5

[0198] This example provides a method for preparing compound I a9 -1, the structure of which is as follows: a9

[0199]

[0200] The method for preparing starting compound Cy3-CHO-OCH3-CH3-CH2SO3is similar to that for preparing Cy3-CHO-1, except that different starting materials are used.

[0201]

[0202] Compound I a9 -1, specifically comprising the following steps:

[0203] Compound Cy3-CHO-OCH3-CH3-CH2SO3(0.275 g, 0.5 mmol) and cyclohexylamine (0.099 g, 1 mmol) were dissolved in MeOH (6 mL), 2 drops of DIPEA were added dropwise, and the mixture was stirred at 45-55°C under a nitrogen atmosphere overnight. After the reaction was completed, KBH4(0.54 g, 0.5 mmol) was added to the mixture for reduction, the solvent was evaporated, the mixture was extracted with DCM / H2O, DDQ (0.036 g, 0.25 mmol) was added to the CH2Cl2solution for oxidation, and the mixture was purified by column chromatography (MeOH / DCM) to obtain compound I a9 -1. The structure of compound I a9 -1 is characterized as follows: HRMS-ESI: m / z calcd M + for C 37 H 51 SN3O4 + ,633.8920found,633.8923.

[0204] Example 6

[0205] This example provides a method for preparing compound I a17 -1, the structure of which is as follows: a17

[0206]

[0207] ​​The preparation method of the starting compound Cy3-CHO--ph-H-COOH having the following structure is similar to that of Cy3-CHO-1, except that different initial starting material is used;

[0208]

[0209] Compound I a17 The preparation of -1 specifically comprises the following steps:

[0210] Compound Cy3-CHO--ph-H-COOH (0.266 g, 0.5 mmol) and p-aminophenylarsonic acid (0.2 g, 0.8 mmol) were dissolved in MeOH (6 mL), 2 drops of DIPEA were added dropwise, and the mixture was stirred at 45-55°C overnight. After the reaction was completed, KBH4 (0.54 g, 0.5 mmol) was added to the mixture for reduction reaction, the solvent was evaporated, extraction was performed with DCM / H2O, DDQ (0.036 g, 0.25 mmol) was added to the CH2Cl2 solution for oxidation, and purification was performed by column chromatography (MeOH / DCM) to obtain compound I a17 -1. Compound I a17 The structure of -1 is characterized as follows: HRMS-ESI: m / z calcd M + for C 43 H 47 AsS2N3O2 + ,776.9090; found 776.9096.

[0211] Example 7

[0212] This example provides a preparation method of compound I a29 -1. The structure of compound I a17 -1 is as follows:

[0213]

[0214] The preparation method of the starting compound Cy5-CHO-H-phB(OH)2-OH having the following structure is similar to that of Cy5-CHO-1, except that different initial starting material is used;

[0215]

[0216] Compound I a29 -1 is similar to the preparation method of compound I a6 -1 in Example 4, except that compound Cy5-CHO-CH3-ph-OH is replaced by compound Cy5-CHO-H-phB(OH)2--OH, and butylamine is replaced by an acylated phalloidin product.

[0217] Example 8

[0218] This example provides a method for preparing compound I a37 -1, which is shown below: a37 -1, which is shown below:

[0219]

[0220] A method for preparing starting compound Cy3-CHO-I-H-H having the following structure is similar to the method for preparing Cy3-CHO-I, except that a different starting material is used.

[0221]

[0222] Compound I a37 -1, which is shown below: a9 -1, which is shown below:

[0223] Example 9

[0224] This example provides a method for preparing compound I b41 -1, which is shown below: b41 -1, which is shown below:

[0225]

[0226] A method for preparing starting compound Cy5-CHO-Cl-phNO2-H having the following structure is similar to the method for preparing Cy5-CHO-I, except that a different starting material is used.

[0227]

[0228] Compound I b41 -1, which is shown below:

[0229] Compound Cy5-CHO-Cl-phNO2-H (0.259 g, 0.5 mmol) was dissolved in MeOH (6 mL) with 4-methylquinolinium salt (0.158 g, 1 mmol) and 5 drops of piperidine, and the reaction was refluxed at 80 °C for 24 h. After the reaction was completed, the solvent was evaporated, and the product was purified by column chromatography (MeOH / DCM) to obtain compound I b41 -1. Compound I b41 -1, which is shown below: 1H-NMR (400 MHz, DMSO-d6): 9.01 (t, 1H), 8.95 (d, 1H), 8.60 (s, 1H), 8.42 (s, 1H), 8.23 (s, 1H), 8.12 (d, 1H), 8.10 (s, 1H), 7.94 (t, 1H), 7.71 (m, 1H), 7.44 (d, 1H), 7.34 (m, 1H), 7.30 (d, 1H), 6.62 (t, 1H), 4.39 (d, 3H), 7.47 (t, 1H), 6.95 (t, 2H), 3.88 (t, 2H), 3.09 (d, 2H), 1.81 (t, 2H), 1.79 (m, 6H), 1.63 (d, 2H), 1.44 (d, 6H), 1.36 (s, 2H). 13 C-NMR (100 MHz, DMSO-d6): 183.5, 164.4, 157.0, 146.3, 145.8, 141.4, 141.2, 140.8, 139.8, 136.0, 135.2, 133.4, 128.3, 127.9, 126.9, 126.8, 125.0, 124.8, 124.5, 123.9, 123.4, 122.1, 120.1, 118.6, 111.0, 109.6, 107.7, 78.2, 76.8, 52.8, 51.3, 49.0, 45.3, 43.3, 31.3, 27.1, 26.7, 25.0. RMS-ESI: m / z calcd M + for C 42 H 43 ClN4O2 2+ , 671.2809; found, 671.2806.

[0230] Example 10

[0231] This example provides a method for preparing compound II-1, the structure of which is as follows: b51 b51

[0232]

[0233] The specific steps are as follows:

[0234] Compound Cy3B-CHO-1 (0.225 g, 0.5 mmol) was dissolved in MeOH (6 mL) with 1,3,3, trimethylbenzoindole salt (0.223 g, 1 mmol), 5 drops of piperidine was added, and the reaction was carried out at 55-65 °C for 24 h. After the reaction was completed, the solvent was evaporated, and the compound II-1 was purified by column chromatography (MeOH / DCM) to obtain the compound II-1. b51 ​​-1. Compound II b51 The structure of -1 is characterized as follows: 1 H-NMR (400 MHz, DMSO-d6): 8.95 (d, 1H), 8.12 (d, 1H), 8.08 (s, 1H), 8.04 (s, 1H), 7.79 (m, 1H), 7.79 (m, 1H), 7.71 (m, 1H), 7.65 (d, 1H), 7.62 (d, 1H), 7.44 (d, 1H), 7.34 (m, 1H), 7.32 (d, 1H), 6.79 (d, 1H), 6.62 (m, 1H), 5.67 (d, 1H), 4.07 (d, 2H), 3.88 (t, 2H), 3.09 (d, 2H), 1.81 (t, 2H), 1.79 (m, 6H), 1.63 (d, 2H), 1.44 (d, 6H), 1.57 (d, 6H), 1.41 (m, 3H). 13 C-NMR (100 MHz, DMSO-d6): 167.5, 164.6, 164.4, 152.8, 150.1, 145.8, 141.0, 140.8, 140.1, 135.2, 129.5, 127.9, 127.2, 126.9, 126.8, 124.5, 123.3, 120.0, 115.1, 112.2, 109.6, 107.7, 77.9, 75.3, 58.0, 57.1, 56.0, 53.6, 51.3, 43.3, 34.2, 30.7, 30.0, 29.0, 26.7, 24.8, 27.1. HRMS-ESI: m / z calcd M + forC 47 H 49 N3O + C 43 H 47 N3O + ,671.9297 found,671.9293.

[0235] Example 11

[0236] This example provides a method for preparing compound II a5 -1. Compound II a5 The structure of -1 is as follows:

[0237]

[0238] The method for preparing starting compound Cy3B-CHO-ph is similar to that of Cy3B-CHO-1, except that different initial starting materials are used;

[0239]

[0240] Compound II a5 -1, specifically comprising the following steps:

[0241] Cy3B-CHO-ph (0.263 g, 0.5 mmol) and pentylamine (0.087 g, 1 mmol) were dissolved in MeOH (6 mL), 2 drops of DIPEA were added, stirred at 45-55°C under nitrogen atmosphere overnight, after the reaction was completed, KBH4 (0.54 g, 0.5 mmol) was added to the mixture for reduction reaction, the solvent was evaporated, extracted with DCM / H2O, DDQ (0.036 g, 0.25 mmol) was added to the CH2Cl2 solution for oxidation, and purified by column (MeOH / DCM) to obtain compound II a5 -1. Compound II a5 The structure of compound II-1 is characterized as follows: 1 H-NMR (400MHz, DMSO-d6): 9.14 (m, 1H), 8.06 (m, 1H), 7.77 (m, 1H), 7.75 (d, 2H), 7.75 (d, 2H), 7.41 (m, 1H), 7.41 (d, 1H), 7.37 (m, 1H), 7.18 (m, 1H), 6.70 (m, 1H), 4.16 (t, 1H), 3.88 (t, 2H), 3.64 (t, 2H), 3.09 (d, 2H), 2.53 (d, 2H), 1.81 (t, 2H), 1.79 (m, 6H), 1.63 (d, 2H), 1.44 (t, 6H), 1.38 (t, 2H), 1.36 (s, 2H), 1.29 (t, 2H), 1.28 (t, 2H), 0.88 (d, 3H). 13 C-NMR (100MHz, DMSO-d6): 183.5, 164.4, 148.6, 144.9, 141.4, 141.2, 140.7, 140.3, 135.2, 129.2, 127.9, 127.7, 125.9, 125.0, 124.5, 109.6, 78.2, 76.8, 53.1, 52.9, 51.2, 49.5, 49.0, 43.3, 31.3, 30.2, 29.2, 26.7, 27.1, 25.0, 22.4, 14.1. HRMS-ESI: m / z calcd M + for C 41 H 48 N3O + ,598.8545 found,598.8540.

[0242] Example 12

[0243] The present example provides a compound II a15 -1, and a preparation method of the compound II a15 The structure of the compound II

[0244]

[0245] A preparation method of a starting compound Cy3B-CHO-Cl having the following structure is similar to the preparation method of Cy3B-CHO-1, except that different initial starting materials are used.

[0246]

[0247] The compound II a5 -1, and a preparation method of the compound II

[0248] Cy3B-CHO-Cl (0.240 g, 0.5 mmol) and 4-aminophenol (0.109 g, 1 mmol) were added to MeOH (6 mL), 2 drops of DIPEA were added dropwise, and stirring was performed at 45-55°C under a nitrogen atmosphere overnight. After the reaction was completed, KBH4 (0.54 g, 0.5 mmol) was added to the mixture to perform a reduction reaction, the solvent was evaporated, extraction was performed using DCM / H2O, DDQ (0.036 g, 0.25 mmol) was added to the CH2Cl2solution to perform oxidation, and purification was performed by a column (MeOH / DCM) to obtain the compound II a15 -1. The compound II a15 The structure of the compound II 1 H-NMR (400 MHz, DMSO-d6): 9.44 (s, 1H), 8.95 (d, 1H), 8.12 (d, 1H), 7.47 (m, 1H), 7.44 (d, 1H), 7.37 (m, 1H), 7.35 (t, 1H), 7.30 (d, 1H), 7.18 (m, 2H), 7.08 (d, 1H), 6.83 (d, 3H), 6.70 (m, 1H), 6.67 (d, 1H), 4.20 (t, 2H), 3.88 (t, 1H), 3.09 (d, 2H), 1.81 (t, 2H), 1.79 (m, 6H), 1.63 (d, 2H), 1.44 (t, 6H), 1.36 (s, 2H). 13C-NMR(100MHz,DMSO-d6):183.5,164.4,149.3,144.6,141.6,141.2,140.7,135.2,129.7,129.5,128.3,127.9,126.7,125.0,1 24.9,120.8,120.1,113.5,113.0,111.0,109.6,78.2,76.8,52.8,51.2,49.0,48.3,43.3,31.3,27.1,26.7,25.0.HRMS-ESI:m / z calcdM + for C 36 H 37 N3ClO2 + ,579.1605found,579.1608.

[0249] Example 13

[0250] This embodiment provides compound IIb. 46 Preparation method of -1, compound IIb 46 The structure of -1 is as follows:

[0251]

[0252] The preparation method of the starting compound Cy3B-CHO-CH3 with the following structure is similar to that of Cy3B-CHO-1, except that different starting materials are used.

[0253]

[0254] Compound IIb 46 The preparation of -1 includes the following steps:

[0255] Compound Cy3B-CHO-CH3 (0.233 g, 0.5 mmol) and 4-methylquinoline salt (0.158 g, 1 mmol) were reacted in MeOH (6 mL) solution with 5 drops of piperidine. The mixture was refluxed at 80 °C for 24 h. After the reaction was completed, the solvent was evaporated, and the mixture was purified by column chromatography (MeOH / DCM) to obtain compound IIb. 46 -1. Compound IIb 46 The structural characterization of -1 is as follows: 1H-NMR (400 MHz, DMSO-d6): 9.01 (t, 1H), 8.95 (d, 1H), 8.60 (s, 1H), 8.42 (s, 1H), 8.23 (s, 1H), 8.12 (d, 1H), 8.10 (s, 1H), 7.94 (t, 1H), 7.71 (m, 1H), 7.44 (d, 1H), 7.34 (m, 1H), 7.30 (d, 1H), 6.62 (t, 1H), 4.39 (d, 3H), 7.47 (t, 1H), 6.95 (t, 2H), 3.88 (t, 2H), 3.09 (d, 2H), 1.81 (t, 2H), 1.79 (m, 6H), 1.63 (d, 2H), 1.44 (d, 6H), 1.36 (s, 2H). 13 C-NMR (100 MHz, DMSO-d6): 183.5, 164.4, 157.0, 146.3, 145.8, 141.4, 141.2, 140.8, 139.8, 136.0, 135.2, 133.4, 128.3, 127.9, 126.9, 126.8, 125.0, 124.8, 124.5, 123.9, 123.4, 122.1, 120.1, 118.6, 111.0, 109.6, 107.7, 78.2, 76.8, 52.8, 51.3, 49.0, 45.3, 43.3, 31.3, 27.1, 26.7, 25.0. RMS-ESI: m / z calcd M + for C 42 H 43 N3O 2+ , 605.8249 found, 606.8244.

[0256] Example 14

[0257] This example provides a method for preparing compound I b53 -1. The structure of compound I b53 -1 is as follows:

[0258]

[0259] The method for preparing starting compound Cy3-CHO-phOCH3-O-CH2CCH is similar to that for preparing Cy3-CHO-1, except that different starting materials are used;

[0260]

[0261] The preparation of compound I b53 -1 is similar to that of compound II b51The preparation method of compound II-1 is similar to that of compound II, except that compound Cy3B-CHO-1 is replaced by compound Cy3B-CHO-OCH3, and 1,3,3, trimethylbenzoindole salt is replaced by morpholino group modified naphthylamine. The structure of compound II-1 is as follows:

[0262] Example 15

[0263] This example provides a compound II a26 The preparation method of compound II-1 is similar to that of compound II, except that compound Cy3B-CHO-1 is replaced by compound Cy3B-CHO-OCH3, and 1,3,3, trimethylbenzoindole salt is replaced by morpholino group modified naphthylamine. The structure of compound II-1 is as follows: a26

[0264]

[0265] The preparation method of starting compound Cy3B-CHO-I with the following structure is similar to that of compound Cy3B-CHO-1, except that different initial starting materials are used.

[0266]

[0267] Compound II a26 The preparation of compound II-1 is similar to that of compound II in Example 14. b51 The preparation method of compound II-1 is similar to that of compound II, except that compound Cy3B-CHO-1 is replaced by compound Cy3B-CHO-OCH3, and 1,3,3, trimethylbenzoindole salt is replaced by morpholino group modified naphthylamine. The structure of compound II-1 is as follows: a26 -1.HRMS-ESI: m / z calcd for C + for C 47 H 53 N4O4 + ,737.9645found,737.9641.

[0268] Example 16

[0269] This example provides a compound II a29 The preparation method of compound II-1 is similar to that of compound II, except that compound Cy3B-CHO-1 is replaced by compound Cy3B-CHO-OCH3, and 1,3,3, trimethylbenzoindole salt is replaced by morpholino group modified naphthylamine. The structure of compound II-1 is as follows: a29

[0270]

[0271] The preparation method of starting compound Cy3B-CHO-I with the following structure is similar to that of compound Cy3B-CHO-1, except that different initial starting materials are used.

[0272]

[0273] Compound II a29 The preparation of compound II-1 is similar to that of compound II in Example 14.​​b51 The preparation method for -1 is similar, except that compound Cy3B-CHO-1 is replaced with compound Cy3B-CHO-I, and 1,3,3,trimethylbenzoindole salt is replaced with amidated phalloidin peptide product.

[0274] Application examples

[0275] The preparation of biological samples is to achieve specific labeling of target structures and obtain high-resolution and high signal-to-noise ratio imaging results. This invention uses live cells (macrophages) as biological samples and employs cell staining labeling for imaging experiments. Among them, the macrophages used in the test include HeLa cells, COS-7 cells, and 4T1 cells. HeLa cells are derived from Henrietta Lacks' cervical cancer cell line. COS-7 cells are derived from African green monkey kidney fibroblasts and transformed by the SV40 virus gene. 4T1 cells are derived from mouse breast cancer cells.

[0276] Live cell imaging assay, the assay process is as follows:

[0277] Resuscitate and passage macrophages according to standard experimental methods, specifically including:

[0278] Cell thawing: Before thawing, thaw the frozen cells and culture medium in a water bath at 37°C. On a clean work surface, transfer the thawed cell suspension to a centrifuge tube, add culture medium, and centrifuge at 1500 rpm for 5 minutes. Discard the supernatant, add more culture medium to resuspend the cells at the bottom, and gently mix. Transfer the well-mixed cell solution to a cell culture flask and place it in a cell culture incubator (37°C, 5% CO2). Incubate for 24 hours, then change the medium or perform passage according to the cell density.

[0279] Cell passage: When the cell density reaches 80% or higher under a microscope, cell passage can be performed. First, wash the cells with PBS solution 1-2 times, add trypsin to digest them, blow off the adherent cells, centrifuge the cell solution, mix well, and transfer to a culture flask.

[0280] Cell cryopreservation: The procedure for cell cryopreservation is basically the same as that for cell passage, except that after centrifugation, the supernatant is removed, cryopreservation solution is added, the mixture is homogenized, transferred to cryovials, and stored in liquid nitrogen. The cell cryopreservation solution is prepared by mixing DMSO, FBS, and cell culture medium in a ratio of 1:2:7.

[0281] After the standard experimental method of resuscitation and subculture of good macrophages, prepare multiple cell culture media containing resuscitated and subcultured good macrophages, add DMSO solutions (concentration of 0.5 μM) of compounds Cy3B-1, compounds Ia6-1, Ia9-1, Ia29-1, Ia37-1, IIa5-1, IIa46-1, IIa53-1 to the culture medium respectively, after 30 minutes of culture at 37℃, 5% carbon dioxide concentration, place under laser confocal microscope, select excitation wavelength 561 nm, collect fluorescence above 575 nm, when STED super-resolution imaging is performed, the excitation wavelength is 561 nm, the loss light is 775 nm, the fluorescence above 575 nm is collected, and the confocal and STED imaging of living cells in the culture medium is observed; under the CLSM and STED two collection modes, the imaging results of the same target structure are obtained, Figure 1 STED super-resolution microscopic imaging diagram corresponding to compound CyB (the picture corresponds to COS-7 cells) Figure 2 COS-7 cell STED super-resolution microscopic imaging diagram corresponding to compound Ia6-1. Figure 3 COS-7 cell STED super-resolution microscopic imaging diagram corresponding to compound Ia9-1. Figure 4 Hala cell local microfilament microtubule STED super-resolution microscopic imaging diagram corresponding to compound Ia29-1. Figure 5 COS-7 cell local mitochondria STED super-resolution microscopic imaging diagram corresponding to compound Ia37-1. Figure 6 COS-7 cell mitochondria STED super-resolution microscopic imaging diagram corresponding to compound IIa5-1. Figure 7 Cell local mitochondria STED super-resolution microscopic imaging diagram corresponding to compound IIa46-1. Figure 8 Cell local mitochondria STED super-resolution microscopic imaging diagram corresponding to compound IIa53-1.

[0282] The probe can realize live cell staining within 5-10 min, which fully proves that it has good biocompatibility and excellent cell permeability. Based on Cy3B-1 dye for live cell imaging, the confocal imaging and STED super-resolution imaging effect of Cy3B-1 is poor, and only the dye entering the mitochondria can be vaguely seen, and the specific fine structure cannot be observed; the image structure of example compounds Ia6-1, Ia9-1, Ia29-1, Ia37-1, IIa5-1, IIa46-1, IIa53-1 shows that mitochondria can show more detailed information under the condition of STED super-resolution imaging than CLSM, the structure of mitochondrial cristae and the microfilament microtubules on the cytoskeleton can be seen, and the contrast is obviously improved, the structure of the cristae is very clear, the STED super-resolution imaging of mitochondria can be realized, and it has the performance of super-resolution imaging, realizes the adjustment of spectral performance, improves the light stability of the dye, and shows excellent performance such as low phototoxicity.

[0283] The STED super-resolution imaging effect of the corresponding cells of compounds Ia17-1, Ib41-1, IIb51-1, Ib53-1 IIa26-1, IIa29-1 can also observe the structure of mitochondrial cristae, which also has good super-resolution effect.

[0284] Obviously, the above examples are only examples for the purpose of clarity, and are not limitations on the embodiments. Based on the above description, other different forms of changes or variations can also be made by those of ordinary skill in the art. All embodiments do not need and cannot be exhausted here. The obvious changes or variations derived therefrom are still within the protection scope of the present application.

Claims

1. A fluorescent dye compound, characterized in that, The fluorescent dye compound has one of the following structures: ； S1 is selected from one of the following groups: S2 is selected from one of the following groups: Z － Selected from Cl - ,Br - I - OTs － ClO4 - PF6 - ; R1 is selected from any one of hydrogen, unsubstituted C1-5 alkoxy, or unsubstituted C6-C15 aryl.

2. The fluorescent dye compound according to claim 1, characterized in that, The fluorescent dye compound has the following structure: Among them, R1, Z - The definition is the same as that in claim 1.

3. The use of the fluorescent dye compound according to claim 1 or 2 in the preparation of live-cell fluorescent imaging agents and super-resolution imaging agents.

Images