CN107141840A - Novel near-infrared high-quantum-yield dye and preparation and application thereof - Google Patents

Abstract

The invention discloses a novel near-infrared high quantum yield dye, which has the following structural formula I: R ₁ , R ₂ , R ₆ , and R ₇ are independently hydrogen, lower hydrocarbon group, ether group, substituted alkyl group or acyl group; R ₃ , R ₄ , R ₅ , R ₈ , R ₉ , and R ₁₀ are independently Hydrogen, lower alkyl or halogen; R ₁₁ is hydrogen, methyl, cyano, or trifluoromethyl; It is an anion; the preparation method of the dye is also disclosed. The dye has good biocompatibility, low toxicity, long fluorescence emission and high fluorescence quantum yield, avoids background autofluorescence, and can obtain fluorescence imaging images with high signal-to-noise ratio, and is used for nucleoli in biological systems Fluorescent labeling of RNA plays an important role in the study of physiological processes related to nucleolar RNA.

Description

A new near-infrared dye with high quantum yield and its preparation and application

technical field

The invention relates to a near-infrared nucleolus RNA dye, and provides a novel near-infrared dye with high quantum yield, preparation and application.

Background technique

Owing to its high spatiotemporal resolution and fast and non-invasive operation, small molecule fluorescent dye-based fluorescence imaging has become a powerful tool for visual observation of biological phenomena (such as biomolecular detection, labeling, cancer diagnosis and treatment, etc.) in living systems. . RNA is a particularly important biomolecule in living systems, and RNA in the nucleus is located in the nucleolus, where they play crucial roles in gene coding, transcription, and expression. Many physiological processes in biochemical processes, including cell proliferation and differentiation, are closely related to the function of RNA. Therefore, real-time imaging of RNA is of great significance in biochemical research. Many fluorescent imaging techniques (RNA microinjection, fluorescence in situ hybridization (FISH) or labeling of green fluorescent protein (GFP) and its derivative yellow fluorescent protein (YFP)) have been utilized to visualize RNA in living cells, but these techniques have Poor cell permeability and may affect normal cell function. Therefore, the development of small molecule dyes with good photophysical properties, high cell penetration and easy manipulation is of great significance for RNA selective imaging.

So far, Invitrogen's RNASelect with green fluorescence emission is the only commercial RNA probe for cell imaging, but its application is not very wide, the dyeing selectivity is poor, and the fluorescence wavelength is relatively short. The molecular structure and The method of synthesis has not been disclosed so far. In recent years, researchers have developed many dyes for RNA labeling through various efforts. In most cases, obtaining a satisfactory image of RNA in living cells requires a long incubation time (tens of minutes or even hours), and only a few positively charged dyes can smoothly enter cells and rapidly image RNA , and the reported dye incubation concentration is high and the quantum yield is also low, and these high concentration dyes may affect the normal physiological activities of living cells and interfere with the background fluorescence in the cells. Importantly, RNA dyes emitting deep red, especially near-infrared, are in high demand due to minimal photodamage, minimal background fluorescence interference, reduced Light scattering and enhanced tissue penetration depth. Therefore, it is of great significance to develop new RNA dyes with rapid localization, low incubation concentration, high quantum yield, high photostability and near-infrared emission.

Compared with commonly used dyes (coumarin, naphthalene diimide, BODIPY, cyanine dyes, etc.), xanthene-based fluorescent dyes have attracted much attention due to their unique properties: long wavelength, good photostability, and high quantum yield , large molar extinction coefficient, simple synthesis, easy modification, etc., are widely used in the field of biomedicine. However, its short emission wavelength and strong pH sensitivity limit its application in the in vivo field. The new long-wavelength xanthene fluorescent dyes reported in recent years are mainly red-shifted by expanding the electronic conjugate system and introducing electron-withdrawing groups. However, it is very difficult to obtain structures with longer emission and high fluorescence quantum yield.

Contents of the invention

The first technical problem to be solved by the present invention is to provide a novel near-infrared dye with high quantum yield. This dye has good biocompatibility, low toxicity, long fluorescence emission wavelength, excellent chemical stability and high fluorescence quantum yield.

The second technical problem to be solved by the present invention is to provide a method for preparing a novel near-infrared dye with high quantum yield.

The third technical problem to be solved by the present invention is to provide a new near-infrared dye with high quantum yield as a near-infrared nucleolar RNA dye.

In order to solve the first technical problem, a novel near-infrared high quantum yield dye of the present invention is characterized in that it has the following structural formula I:

R ₁ , R ₂ , R ₆ , and R ₇ are independently hydrogen, lower hydrocarbon group, ether group, substituted alkyl group or acyl group; R ₃ , R ₄ , R ₅ , R ₈ , R ₉ , and R ₁₀ are independently Hydrogen, lower alkyl or halogen; R ₁₁ is hydrogen, methyl, cyano, or trifluoromethyl; is an anion;

The lower hydrocarbon groups in R ₁ , R ₂ , R ₆ , and R ₇ are linear, branched or cyclic; the lower hydrocarbon groups contain 1-6 carbon atoms;

The number of carbon atoms in the ether group is 3-6, and the number of oxygen atoms is ≤2;

The substituted alkyl group is a straight chain or branched chain; the substituted alkyl group is ω-formate group substituted 1-6 carbon atom alkyl, ω-formamide group substituted 1-6 carbon atom alkyl, ω -Halogen substituted 1-6 carbon atom alkyl, ω-hydroxyl substituted 1-6 carbon atom alkyl, ω-amino substituted 1-6 carbon atom alkyl or ω-mercapto substituted 1-6 carbon atom alkyl ; Wherein, the formate group in the ω-formyl group substituting 1-6 carbon atom alkyl groups is a 2-5 carbon atom alkyl formate group: the ω-formamide group replaces 1- The carboxamide group in the 6 carbon atom alkyl group is a 2-5 carbon atom hydrocarbon group carboxamide group;

The acyl group is an alkylacyl group with 2-6 carbon atoms, a tert-butoxycarbonyl group, a benzoyl group, a benzoyl group with 1-6 carbon atoms or a halogen-substituted benzoyl group;

The halogen is fluorine, chlorine, bromine or iodine;

The lower hydrocarbon group in R ₃ , R ₄ , R ₅ , R ₈ , R ₉ , R ₁₀ is methyl or ethyl;

said is any organic or inorganic anion that balances the charge.

Further, in the structural formula I, R ₁ and R ₃ , R ₁ and R ₁₀ , R ₂ and R ₃ , R ₂ and R ₁₀ , R ₆ and R ₅ , R ₆ and R ₈ , R ₇ and R ₅ , R ₇ and R ₈ , R ₁ and R ₂ or R ₆ and R ₇ can independently form the following Ia-In structure:

Wherein R is hydrogen, methyl or ethyl; Y is C, O or NR.

Further, the lower hydrocarbon groups in R ₁ , R ₂ , R ₆ , and R ₇ are methyl, ethyl, propyl, isopropyl, cyclopropyl, allyl, butyl, isobutyl or tert-butyl;

The ether group is CH ₂ CH ₂ OCH ₃ , CH ₂ CH ₂ OCH ₂ CH ₃ , CH ₂ CH ₂ OCH ₂ CH ₃ OH, CH ₂ CH ₂ OCH ₂ CH ₃ OCH ₃ or CH ₂ CH ₂ OCH ₂ CH _{3 OCH2CH3 ;}

The omega-2-5 carbon atom alkyl formate group is substituted with 1-6 carbon atom alkyl group as (CH ₂ ) _m COO(CH ₂ ) _n CH ₃ or (CH ₂ ) _m COOC(CH ₃ ) ₃ , where m is 1-4, n is 0-3;

The omega-2-5 carbon atom hydrocarbyl carboxamide group substituting 1-6 carbon atom alkyl group is (CH ₂ ) _m CONH(CH ₂ ) _n CH ₃ or (CH ₂ ) _m CON[(CH ₂ ) _n CH ₃ ] ₂ , wherein m is 1-4, n is 0-3;

The ω-halogen substituted alkyl group with 1-6 carbon atoms is (CH ₂ ) _m Cl, (CH ₂ ) _m Br or (CH ₂ ) _m I, wherein m is 1-6;

The ω-hydroxyl substituted alkyl group with 1-6 carbon atoms is (CH ₂ ) _m OH, where m is 1-6;

The ω-amino group substituted with an alkyl group of 1-6 carbon atoms is (CH ₂ ) _m NH ₂ , where m is 1-6;

The ω-mercapto substituted alkyl group with 1-6 carbon atoms is (CH ₂ ) _m SH, where m is 1-6;

The 2-6 carbon atom alkyl acyl group is acetyl, propionyl, butyryl or tert-butyryl;

The 1-6 carbon atom substituted benzoyl is methyl substituted benzoyl, ethyl substituted benzoyl, propyl substituted benzoyl, butyl substituted benzoyl or tert-butyl substituted benzoyl;

The halogen-substituted benzoyl is chlorine-substituted benzoyl, bromo-substituted benzoyl or iodine-substituted benzoyl.

_In order to solve the second technical problem, the present invention provides the preparation method of the high quantum yield dye of novel near-infrared, when R in structural formula I When being hydrogen or trifluoromethyl, comprise following preparation steps:

Add 1 mmol of compound II and 1 mmol of compound III to 5-15 ml of concentrated sulfuric acid, react under heating for 1-4 hours, and slowly add the reaction solution dropwise to 20-100 times the volume of ice water after cooling. Then add 0.01-1 times volume of perchloric acid with a mass percent concentration of 70%, stir well, precipitate solid, filter, vacuum dry, and separate the product by column chromatography.

Wherein, the heating temperature is 60-100°C;

The structural formula of the compound II is as follows:

The structural formula of the compound III is as follows:

Among them, R ₁ , R ₂ , R ₆ , and R ₇ are independently hydrogen, lower hydrocarbon group, ether group, substituted alkyl group or acyl group; R ₃ , R ₄ , R ₅ , R ₈ , R ₉ , and R ₁₀ are independently R is hydrogen, lower alkyl or halogen _; R is hydrogen or trifluoromethyl.

Or, when R in the structural _formula I is a methyl group, the following preparation steps are included:

1) Add 1 mmol of compound II and 1 mmol of compound III to 5-15 ml of concentrated sulfuric acid, react for 1-4 hours under heating conditions, and slowly add 20-100 times the volume of ice to the reaction solution after cooling Add 0.01-1 volume of perchloric acid with a mass percentage concentration of 70% to the water, stir well, precipitate solid, filter, vacuum dry, and separate by column chromatography to obtain product 1-a;

2) Dissolve 1 mmol of Ia obtained in 5 ml of anhydrous THF, add dropwise 3-6 ml of THF solution containing 1 mmol of CH ₃ MgI under N ₂ protection conditions, stir at room temperature for 24 hours, and then add 2-5 ml of a THF solution containing 1 mmol DDQ was continued to stir for 1-12 h, the reaction solution was spin-dried and separated by column chromatography to obtain the product.

Wherein, the heating temperature is 60-100°C;

The structural formula of the compound II is as follows:

The structural formula of the compound III is as follows:

Among them, R ₁ , R ₂ , R ₆ , and R ₇ are independently hydrogen, lower hydrocarbon group, ether group, substituted alkyl group or acyl group; R ₃ , R ₄ , R ₅ , R ₈ , R ₉ , and R ₁₀ are independently R is hydrogen, lower hydrocarbon or halogen; R ₀ is hydrogen.

Or, when R in the structural formula I ₁₁ is a cyano group, comprising the following preparation steps:

① Add 1 mmol of compound II and 1 mmol of compound III to 5-15 ml of concentrated sulfuric acid, react for 1-4 hours under heating conditions, and slowly add the reaction solution dropwise to 20-100 times the volume of ice water after cooling , then add 0.01-1 times the volume of perchloric acid with a mass percentage concentration of 70%, stir well, precipitate solids, filter, vacuum dry, and separate by column chromatography to obtain product 1-a;

②Dissolve 1 mmol of I-a obtained in 5-20 ml of anhydrous DMF, then add 1 mmol of NaCN, stir at room temperature for 24 hours, then add 2-5 ml of THF solution containing 1 mmol of DDQ, and react The product was separated by column chromatography after spinning to dryness.

Wherein, the heating temperature is 60-100°C;

The structural formula of the compound II is as follows:

The structural formula of the compound III is as follows:

Among them, R ₁ , R ₂ , R ₆ , and R ₇ are independently hydrogen, lower hydrocarbon group, ether group, substituted alkyl group or acyl group; R ₃ , R ₄ , R ₅ , R ₈ , R ₉ , and R ₁₀ are independently R is hydrogen, lower hydrocarbon or halogen; R ₀ is hydrogen.

Further, in the above three preparation methods, the lower hydrocarbon group in R ₁ , R ₂ , R ₆ , and R ₇ is linear, branched or cyclic; the lower hydrocarbon group contains 1-6 carbon atoms;

The number of carbon atoms in the ether group is 3-6, and the number of oxygen atoms is ≤2;

The substituted alkyl group is a straight chain or branched chain; the substituted alkyl group is ω-formate group substituted 1-6 carbon atom alkyl, ω-formamide group substituted 1-6 carbon atom alkyl, ω -Halogen substituted 1-6 carbon atom alkyl, ω-hydroxyl substituted 1-6 carbon atom alkyl, ω-amino substituted 1-6 carbon atom alkyl or ω-mercapto substituted 1-6 carbon atom alkyl ; Wherein said ω-formyl formate group replaces 1-6 carbon atom alkyl formate group is 2-5 carbon atom alkyl formate group; described ω-formamide group replaces 1-6 The carboxamide group in the carbon atom alkyl group is 2-5 carbon atom hydrocarbon group carboxamide group;

The acyl group is an alkylacyl group with 2-6 carbon atoms, a tert-butoxycarbonyl group, a benzoyl group, a benzoyl group with 1-6 carbon atoms or a halogen-substituted benzoyl group;

The halogen is fluorine, chlorine, bromine or iodine;

The lower hydrocarbon group in R ₃ , R ₄ , R ₅ , R ₈ , R ₉ , and R ₁₀ is methyl or ethyl.

Further, the compound II and compound III can independently form the following IIa-IIg and IIIa-IIIg structures:

Wherein R is hydrogen, methyl or ethyl; Y is C, O or NR.

Preferably, the lower hydrocarbon group in R ₁ , R ₂ , R ₆ , and R ₇ is methyl, ethyl, propyl, isopropyl, cyclopropyl, allyl, butyl, isobutyl or tert Butyl;

Preferably, the ether group is CH ₂ CH ₂ OCH ₃ , CH ₂ CH ₂ OCH ₂ CH ₃ , CH ₂ CH ₂ OCH ₂ CH ₃ OH, CH ₂ CH ₂ OCH ₂ CH ₃ OCH ₃ or CH ₂ CH ₂ OCH _{2CH3OCH2CH3 ; _ _}

Preferably, the ω-2-5 carbon atom alkyl formate group is substituted with 1-6 carbon atom alkyl group as (CH ₂ ) _m COO(CH ₂ ) _n CH ₃ or (CH ₂ ) _m COOC( CH ₃ ) ₃ , wherein m is 1-4, n is 0-3;

Preferably, the ω-2-5 carbon atom hydrocarbyl carboxamide group substituting 1-6 carbon atom alkyl group is (CH ₂ ) _m CONH(CH ₂ ) _n CH ₃ or (CH ₂ ) _m CON[(CH ₂ ) _n CH ₃ ] ₂ , wherein m is 1-4, n is 0-3;

Preferably, the ω-halogen substituted alkyl group with 1-6 carbon atoms is (CH ₂ ) _m Cl, (CH ₂ ) _m Br or (CH ₂ ) _m I, wherein m is 1-6;

Preferably, the ω-hydroxyl is substituted with an alkyl group having 1-6 carbon atoms as (CH ₂ ) _m OH, where m is 1-6;

Preferably, the ω-amino group substituting an alkyl group with 1-6 carbon atoms is (CH ₂ ) _m NH ₂ , where m is 1-6;

Preferably, the ω-mercapto substituted alkyl group with 1-6 carbon atoms is (CH ₂ ) _m SH, where m is 1-6;

Preferably, the 2-6 carbon atom alkyl acyl group is acetyl, propionyl, butyryl or tert-butyryl;

Preferably, the 1-6 carbon atom substituted benzoyl is methyl substituted benzoyl, ethyl substituted benzoyl, propyl substituted benzoyl, butyl substituted benzoyl or tert-butyl substituted benzoyl Acyl;

Preferably, the halogen-substituted benzoyl is chlorine-substituted benzoyl, bromo-substituted benzoyl or iodine-substituted benzoyl.

To solve the third technical problem, the present invention protects the application of a novel near-infrared high quantum yield dye as a near-infrared nucleolar RNA dye.

Furthermore, the present invention protects a novel near-infrared dye with high quantum yield as a near-infrared nucleolar RNA dye, which is applied to the fluorescent labeling of nucleolar RNA.

Some new anticancer drugs, such as cisplatin, actinomycin D, α-amanitin, etc., act on RNA polymerase, thereby inducing cell death, and among RNA polymerases, RNA polymerase I and RNA polymerase III is located in the cytoplasm, and RNA polymerase II is located in the nucleus, so the dyes in the present invention can be used to screen anticancer drugs at different sites of action and the efficacy of anticancer drugs by fluorescently labeling nucleolar RNA evaluation of.

The present invention is based on the readily available 3,4-dihydro-1-naphthone derivatives, through heating the corresponding salicylaldehyde derivatives under the condensation conditions of concentrated sulfuric acid to obtain a series of asymmetric, nonlinear co- A novel xanthene-derived compound with a conjugated structure, which has a longer fluorescence emission (near-infrared fluorescence), especially the methylene structure introduced in the structure makes it have a higher fluorescence quantum yield. In addition, its good biocompatibility and low toxicity allow fluorescent labeling of macromolecules (nucleolar RNA) in biological systems, laying an important foundation for studying its role in physiological activities.

The beneficial effects of the present invention are as follows:

The novel near-infrared high quantum yield dye with structural formula I of the present invention has an asymmetric, non-linear conjugated structure, and has long fluorescence emission (near-infrared emission) and higher fluorescence quantum yield, and also With good biocompatibility and low toxicity, it can fluorescently label nucleolar RNA, an important biomacromolecule in biological systems, especially cells, which reduces the concentration of dye incubation and avoids the interference of high-concentration dyes on the normal physiological functions of cells. Avoiding the interference of background fluorescence and thus accurately locating biological macromolecules plays an important role in studying the physiological processes related to nucleolar RNA.

Description of drawings

The specific implementation manners of the present invention will be further described in detail below in conjunction with the accompanying drawings.

FIG. 1 shows a picture of A549 cells stained with fluorescent dye I-6 in Example 3 of the present invention.

Fig. 2 shows a picture of A549 cells stained with fluorescent dye I-15 in Example 7 of the present invention.

FIG. 3 shows the staining of A549 cells with fluorescent dye I-29 in Example 15 of the present invention.

Fig. 4 shows a picture of A549 cells stained with fluorescent dye I-30 in Example 16 of the present invention.

FIG. 5 shows the staining of A549 cells with fluorescent dye I-42 in Example 27 of the present invention.

FIG. 6 shows the staining of A549 cells with fluorescent dye I-43 in Example 28 of the present invention.

FIG. 7 shows the staining of A549 cells with fluorescent dye I-54 in Example 37 of the present invention.

FIG. 8 shows the staining of HeLa cells with fluorescent dye I-54 in Example 37 of the present invention.

FIG. 9 shows the staining of A549 cells with fluorescent dye I-78 in Example 40 of the present invention.

FIG. 10 shows the staining of HeLa cells with fluorescent dye I-78 in Example 40 of the present invention.

FIG. 11 shows the staining of HeLa cells with fluorescent dye I-95 in Example 42 of the present invention.

FIG. 12 shows the staining of HeLa cells with fluorescent dye I-191 in Example 54 of the present invention.

FIG. 13 shows the staining of HeLa cells with fluorescent dye I-255 in Example 62 of the present invention.

FIG. 14 shows the staining of HeLa cells with fluorescent dye I-393 in Example 77 of the present invention.

Figure 15 shows the fluorescent dye I-54 in Example 37 of the present invention and the fluorescent dye I-78 in Example 40 respectively and I-78 after digestion with DNase and RNase and compared with commercially available DNA and RNA dyes.

Figure 16 shows the comparison of the cell staining effect of the fluorescent dye I-54 in Example 37 of the present invention and the fluorescent dye I-78 in Example 40 with the commercially available RNA dye SYTO RNASelect.

detailed description

In order to illustrate the present invention more clearly, the present invention will be further described below in conjunction with preferred embodiments and accompanying drawings. Similar parts in the figures are denoted by the same reference numerals. Those skilled in the art should understand that the content specifically described below is illustrative rather than restrictive, and should not limit the protection scope of the present invention.

Example 1:

0.193 g (0.001 mol) of compound II-1 and 0.161 g (0.001 mol) of compound III-1 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 2 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.343g of compound I-1, with a yield of 82%. ESI MS: theoretical calculation m/z: 319.18, actual test m/z: 319.20. λ ^abs. _max /nm = 615nm, λ ^em _max /nm = 638nm, Ф _f = 0.80.

0.418g (0.001mol) of compound I-1, 0.202g (0.001mol) of di-tert-butyl dicarbonate and 0.101g (0.001mol) of triethylamine were added to 10 ml of anhydrous dichloromethane respectively, After stirring for 24 hours at room temperature, the solvent was removed under vacuum, dried and purified by column chromatography to obtain 0.440 g of compound I-2 with a yield of 85%. ESI MS: theoretical calculation m/z: 419.23, actual test m/z: 419.25. λ ^abs. _max /nm = 620nm, λ ^em _max /nm = 645nm, Ф _f = 0.82.

Dissolve 0.518g (0.001mmol) of compound I-2 in 5 ml of anhydrous THF, add dropwise to 3 ml of THF solution containing 1 mmol of CH ₃ MgI under nitrogen protection, react at room temperature for 24 hours, then add 2 ml of CH 3 MgI containing 1 mmol DDQ solution in THF was stirred for 1 h, the solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.223 g of compound I-3 with a yield of 42%. ESI MS: theoretical calculation m/z: 433.25, actual test m/z: 433.20. λ ^abs. _max /nm = 622nm, λ ^em _max /nm = 648nm, Ф _f = 0.85.

Example 2

Compound III-11 was prepared from commercial compound III-1 and ethyl iodide according to literature (Org. Lett., 2011, 13, 6488-6491), with a yield of 50%.

0.217 g (0.001 mol) of compound II-2 and 0.189 g (0.001 mol) of compound III-2 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 2 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.399 g of compound I-4, with a yield of 85%. ESI MS: Theoretical calculation m/z: 371.21, actual test m/z: 371.24. λ ^abs. _max /nm = 635nm, λ ^em _max /nm = 658nm, Ф _f = 0.73.

Dissolve 0.470 g (0.001 mmol) of compound I-4 in 5 ml of anhydrous THF, add dropwise to 3 ml of THF solution containing 1 mmol of CH ₃ MgI under nitrogen protection, react at room temperature for 24 hours, then add 2 ml of CH 3 MgI containing 1 mmol DDQ solution in THF was stirred for 1 h, the solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.198 g of compound I-5 with a yield of 41%. ESI MS: theoretical calculation m/z: 385.23, actual test m/z: 383.27. λ ^abs. _max /nm = 636nm, λ ^em _max /nm = 660nm, Ф _f = 0.75.

Example 3

Compound II-3 was prepared from 7-hydroxy-1-methyl-1,2,3,4-tetrahydroquinoline according to the literature (J.Am.Chem.Soc., 2007,129,9986-9998), the yield 76%. Compound III-3 was prepared from commercial compound III-1 and methyl iodide according to literature (Org. Lett., 2011, 13, 6488-6491), with a yield of 42%.

0.191 g (0.001 mol) of compound II-3 and 0.189 g (0.001 mol) of compound III-3 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 2 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.391 g of compound I-6, with a yield of 88%. ESI MS: Theoretical calculation m/z: 345.20, actual test m/z: 345.26. λ ^abs. _max /nm = 633nm, λ ^em _max /nm = 656nm, Ф _f = 0.78.

Dissolve 0.444g (0.001mmol) of compound I-6 in 5 ml of anhydrous THF, add dropwise to 3 ml of THF solution containing 1 mmol of CH ₃ MgI under nitrogen protection, react at room temperature for 24 hours, then add 2 ml of CH 3 MgI containing 1 mmol DDQ solution in THF was stirred for 1 h, the solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.202 g of compound I-7 with a yield of 44%. ESI MS: theoretical calculation m/z: 359.21, actual test m/z: 359.17. λ ^abs. _max /nm = 634nm, λ ^em _max /nm = 658nm, Ф _f = 0.80.

Example 4

Compound II-4 was synthesized from 7-hydroxyl-1-ethyl-2,2,4-trimethyl-1,2-dihydro Quinoline was prepared with a yield of 71%. Compound III-4 was prepared from commercial compound III-1 and 1-iodoethane according to literature (Org. Lett., 2011, 13, 6488-6491), with a yield of 45%.

0.231 g (0.001 mol) of compound II-4 and 0.217 g (0.001 mol) of compound III-4 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 2 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.446 g of compound I-8, with a yield of 87%. ESI MS: theoretical calculation m/z: 413.26, actual test m/z: 413.32. λ ^abs. _max /nm = 648nm, λ ^em _max /nm = 674nm, Ф _f = 0.76.

Dissolve 0.512g (0.001mmol) of compound I-8 in 5 ml of anhydrous THF, add dropwise to 3 ml of THF solution containing 1 mmol of CH ₃ MgI under nitrogen protection, react at room temperature for 24 hours, then add 2 ml of CH 3 MgI containing 1 mmol DDQ solution in THF was stirred for 1 h, the solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.226 g of compound I-9 with a yield of 43%. ESI MS: theoretical calculation m/z: 359.21, actual test m/z: 359.17. λ ^abs. _max /nm = 650nm, λ ^em _max /nm = 678nm, Ф _f = 0.73.

Example 5

Compound II-5 was prepared from 3-N,N-diethylaminophenol according to literature (J.Am.Chem.Soc., 2007, 129, 9986-9998), with a yield of 75%. Compound III-5 was prepared from commercial compound III-1 and 1-bromo-2-(2-methoxyethoxy)ethane according to literature (Org.Lett., 2011, 13, 6488–6491), yield 30 %.

0.165 g (0.001 mol) of compound II-5 and 0.365 g (0.001 mol) of compound III-5 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 2 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.440 g of compound I-10, with a yield of 74%. ESI MS: Theoretical calculation m/z: 495.29, actual test m/z: 495.23. λ ^abs. _max /nm = 622nm, λ ^em _max /nm = 648nm, Ф _f = 0.80.

Dissolve 0.594g (0.001mmol) of compound I-10 in 5 ml of anhydrous THF, add dropwise to 3 ml of THF solution containing 1 mmol of CH ₃ MgI under nitrogen protection, react at room temperature for 24 hours, then add 2 ml of CH 3 MgI containing The THF solution of 1 mmol DDQ was stirred for 1 h, the solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.255 g of compound I-11 with a yield of 42%. ESI MS: Theoretical calculation m/z: 509.30, actual test m/z: 509.27. λ ^abs. _max /nm = 623nm, λ ^em _max /nm = 650nm, Ф _f = 0.83.

Example 6

Compound II-6 was synthesized from 7-hydroxyl-1-(4-n-butyric acid)-1,2,3,4-tetrahydroquinone according to literature (J.Am.Chem.Soc., 2007,129,9986-9998) Phenol was prepared with a yield of 62%. Compound III-6 was prepared from commercial compound III-1 with 1-chloro-3-bromopropane and methyl iodide respectively according to literature (Org. Lett., 2011, 13, 6488-6491), with a yield of 28%.

0.291 g (0.001 mol) of compound II-6 and 0.215 g (0.001 mol) of compound III-6 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 2 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.393 g of compound I-12, with a yield of 69%. ESI MS: Theoretical calculation m/z: 471.26, actual test m/z: 471.21. λ ^abs. _max /nm = 632nm, λ ^em _max /nm = 658nm, Ф _f = 0.74.

Dissolve 0.570 g (0.001 mmol) of compound I-12 in 5 ml of anhydrous THF, add dropwise to 3 ml of THF solution containing 1 mmol of CH ₃ MgI under nitrogen protection, react at room temperature for 24 hours, then add 2 ml of CH 3 MgI containing 1 mmol DDQ solution in THF was stirred for 1 h, the solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.222 g of compound I-13 with a yield of 38%. ESI MS: Theoretical calculation m/z: 485.28, actual test m/z: 485.32. λ ^abs. _max /nm = 633nm, λ ^em _max /nm = 662nm, Ф _f = 0.76.

Example 7

Compound II-7 was prepared from 3-(1-piperidinyl)phenol according to literature (J.Am.Chem.Soc., 2007, 129, 9986-9998), with a yield of 83%. Compound III-7 was prepared from commercial compound III-1 and 1-chloro-3-bromopropane according to literature (Org. Lett., 2011, 13, 6488-6491), with a yield of 20%.

0.205 g (0.001 mol) of compound II-7 and 0.241 g (0.001 mol) of compound III-7 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 2 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.454 g of compound I-14, with a yield of 89%. ESI MS: theoretical calculation m/z: 411.24, actual test m/z: 411.20. λ ^abs. _max /nm = 630nm, λ ^em _max /nm = 656nm, Ф _f = 0.74.

Dissolve 0.510 g (0.001 mmol) of compound I-14 in 5 ml of anhydrous THF, add dropwise to 3 ml of THF solution containing 1 mmol of CH ₃ MgI under nitrogen protection, react at room temperature for 24 hours, then add 2 ml of CH 3 MgI containing 1 mmol DDQ solution in THF was stirred for 1 h, the solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.220 g of compound I-15 with a yield of 42%. ESI MS: Theoretical calculation m/z: 425.26, actual test m/z: 425.22. λ ^abs. _max /nm = 630nm, λ ^em _max /nm = 660nm, Ф _f = 0.75.

Example 8

Compound II-8 was prepared from 3-(4-morpholinyl)phenol according to literature (J.Am.Chem.Soc., 2007, 129, 9986-9998), with a yield of 83%. Compound III-8 was prepared from commercial compound III-1 and 1,5-dibromopentane according to literature (Org. Lett., 2011, 13, 6488-6491), with a yield of 14%.

0.207 g (0.001 mol) of compound II-8 and 0.229 g (0.001 mol) of compound III-8 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 2 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.400 g of compound I-16 with a yield of 80%. ESI MS: theoretical calculation m/z: 401.22, actual test m/z: 401.20. λ ^abs. _max /nm = 624nm, λ ^em _max /nm = 652nm, Ф _f = 0.83.

Dissolve 0.500 g (0.001 mmol) of compound I-16 in 5 ml of anhydrous THF, add dropwise to 3 ml of THF solution containing 1 mmol of CH ₃ MgI under nitrogen protection, react at room temperature for 24 hours, then add 2 ml of CH 3 MgI containing The THF solution of 1 mmol DDQ was stirred for 1 h, the solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.200 g of compound I-17 with a yield of 39%. ESI MS: Theoretical calculation m/z: 415.24, actual test m/z: 415.27. λ ^abs. _max /nm = 621nm, λ ^em _max /nm = 654nm, Ф _f = 0.84.

Example 9

Compound II-9 was prepared from 3-(1-piperazine)phenol according to literature (J.Am.Chem.Soc., 2007, 129, 9986-9998), with a yield of 68%. Compound III-9 was prepared from commercial compound III-1 and 1-bromo-2-(2-bromoethoxy)ethane according to literature (Org. Lett., 2011, 13, 6488-6491), with a yield of 11%.

0.206 g (0.001 mol) of compound II-9 and 0.231 g (0.001 mol) of compound III-9 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 2 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.381 g of compound I-18 with a yield of 76%. ESI MS: theoretical calculation m/z: 402.22, actual test m/z: 402.25. λ ^abs. _max /nm = 625nm, λ ^em _max /nm = 653nm, Ф _f = 0.85.

Dissolve 0.501 g (0.001 mmol) of compound I-18 in 5 ml of anhydrous THF, add dropwise to 3 ml of THF solution containing 1 mmol of CH ₃ MgI under nitrogen protection, react at room temperature for 24 hours, then add 2 ml of CH 3 MgI containing 1 mmol DDQ solution in THF was stirred for 1 h, the solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.191 g of compound I-19 with a yield of 37%. ESI MS: theoretical calculation m/z: 416.23, actual test m/z: 416.19. λ ^abs. _max /nm = 624nm, λ ^em _max /nm = 655nm, Ф _f = 0.87.

Example 10

Compound II-10 was prepared from m-aminophenol according to literature (J.Am.Chem.Soc., 2007, 129, 9986-9998), with a yield of 48%.

0.137 g (0.001 mol) of compound II-10 and 0.189 g (0.001 mol) of compound III-3 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 2 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.304 g of compound I-20, with a yield of 78%. ESI MS: Theoretical calculation m/z: 291.15, actual test m/z: 291.20. λ ^abs. _max /nm = 617nm, λ ^em _max /nm = 636nm, Ф _f = 0.82.

0.390g (0.001mol) of compound I-20, 0.202g (0.001mol) of di-tert-butyl dicarbonate and 0.101g (0.001mol) of triethylamine were added to 10 ml of anhydrous dichloromethane respectively, After stirring for 24 hours at room temperature, the solvent was removed under vacuum, dried, and purified by column chromatography to obtain 0.392 g of compound I-21 with a yield of 80%. ESI MS: theoretical calculation m/z: 391.20, actual test m/z: 391.25. λ ^abs. _max /nm = 620nm, λ ^em _max /nm = 644nm, Ф _f = 0.85.

Dissolve 0.490 g (0.001 mmol) of compound I-21 in 5 ml of anhydrous THF, add dropwise to 3 ml of THF solution containing 1 mmol of CH ₃ MgI under nitrogen protection, react at room temperature for 24 hours, then add 2 ml of CH 3 MgI containing 1 mmol DDQ solution in THF was continued to stir for 1 h, the solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.186 g of compound I-22 with a yield of 37%. ESI MS: Theoretical calculation m/z: 405.22, actual test m/z: 405.24. λ ^abs. _max /nm = 620nm, λ ^em _max /nm = 646nm, Ф _f = 0.87.

Example 11

Compound II-10 was prepared from 3-ethylamino-4-methylphenol according to literature (J.Am.Chem.Soc., 2007, 129, 9986-9998), with a yield of 56%.

0.179 g (0.001 mol) of compound II-11 and 0.217 g (0.001 mol) of compound III-4 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 2 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.359 g of compound I-23, with a yield of 78%. ESI MS: theoretical calculation m/z: 361.23, actual test m/z: 361.18. λ ^abs. _max /nm = 623nm, λ ^em _max /nm = 648nm, Ф _f = 0.87.

Dissolve 0.460 g (0.001 mmol) of compound I-23 in 5 ml of anhydrous THF, add dropwise to 3 ml of THF solution containing 1 mmol of CH ₃ MgI under nitrogen protection, react at room temperature for 24 hours, then add 2 ml of CH 3 MgI containing 1 mmol DDQ solution in THF was stirred for 1 h, the solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.166 g of compound I-24 with a yield of 35%. ESI MS: Theoretical calculation m/z: 375.24, actual test m/z: 375.29. λ ^abs. _max /nm = 621nm, λ ^em _max /nm = 650nm, Ф _f = 0.89.

Example 12

Compound II-12 was prepared from 4-chloro-3-diethylaminophenol according to the literature (J.Am.Chem.Soc., 2007,129,9986-9998), with a yield of 56%; Compound III-10 was prepared according to the literature ( Org. Lett., 2011, 13, 6488–6491) was prepared from commercial compound III-1 and bis(2-bromoethyl)amine with a yield of 8%.

0.227 g (0.001 mol) of compound II-11 and 0.230 g (0.001 mol) of compound III-4 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 2 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.375 g of compound I-25, with a yield of 72%. ESI MS: theoretical calculation m/z: 422.20, actual test m/z: 422.23. λ ^abs. _max /nm = 624nm, λ ^em _max /nm = 652nm, Ф _f = 0.83.

Dissolve 0.521g (0.001mmol) of compound I-25 in 5 ml of anhydrous THF, add dropwise to 3 ml of THF solution containing 1 mmol of CH ₃ MgI under nitrogen protection, react at room temperature for 24 hours, then add 2 ml of CH 3 MgI containing 1 mmol DDQ solution in THF was stirred for 1 h, the solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.176 g of compound I-26 with a yield of 33%. ESI MS: Theoretical calculation m/z: 436.22, actual test m/z: 436.19. λ ^abs. _max /nm = 622nm, λ ^em _max /nm = 655nm, Ф _f = 0.85.

Example 13

Dissolve 0.518g (0.001mmol) of compound I-2 in 5ml of anhydrous DMF, then add 1mmol of NaCN, stir at room temperature for 24h, then add 2ml of THF solution containing 1mmol of DDQ, and continue stirring for 1h. The solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.196 g of compound I-27, with a yield of 36%. ESI MS: theoretical calculation m/z: 444.23, actual test m/z: 444.21. λ ^abs. _max /nm = 690nm, λ ^em _max /nm = 720nm, Ф _f = 0.35.

Example 14

Dissolve 0.470 g (0.001 mmol) of compound I-4 in 5 ml of anhydrous DMF, then add 1 mmol of NaCN, stir at room temperature for 24 h, then add 2 ml of THF solution containing 1 mmol of DDQ, and continue stirring for 1 h. The solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.198 g of compound I-28, with a yield of 40%. ESI MS: theoretical calculation m/z: 396.21, actual test m/z: 396.23. λ ^abs. _max /nm = 698nm, λ ^em _max /nm = 732nm, Ф _f = 0.25.

Example 15

Dissolve 0.444 g (0.001 mmol) of compound I-6 in 5 ml of anhydrous DMF, then add 1 mmol of NaCN, stir at room temperature for 24 h, then add 2 ml of THF solution containing 1 mmol of DDQ, and continue stirring for 1 h. The solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.197 g of compound I-29, with a yield of 42%. ESI MS: theoretical calculation m/z: 370.19, actual test m/z: 370.17. λ ^abs. _max /nm = 705nm, λ ^em _max /nm = 730nm, Ф _f = 0.30.

Example 16

Dissolve 0.512 g (0.001 mmol) of compound I-8 in 5 ml of anhydrous DMF, then add 1 mmol of NaCN, stir at room temperature for 24 h, then add 2 ml of THF solution containing 1 mmol of DDQ, and continue stirring for 1 h. The solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.236 g of compound I-30, with a yield of 44%. ESI MS: theoretical calculation m/z: 438.25, actual test m/z: 438.29. λ ^abs. _max /nm = 715nm, λ ^em _max /nm = 750nm, Ф _f = 0.20.

Example 17

Dissolve 0.594 g (0.001 mmol) of compound I-10 in 5 ml of anhydrous DMF, then add 1 mmol of NaCN, stir at room temperature for 24 h, then add 2 ml of THF solution containing 1 mmol of DDQ, and continue stirring for 1 h. The solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.254 g of compound I-31, with a yield of 41%. ESI MS: theoretical calculation m/z: 520.28, actual test m/z: 520.30. λ ^abs. _max /nm = 694nm, λ ^em _max /nm = 612nm, Ф _f = 0.33.

Example 18

Dissolve 0.570 g (0.001 mmol) of compound I-12 in 5 ml of anhydrous DMF, then add 1 mmol of NaCN, stir at room temperature for 24 h, then add 2 ml of THF solution containing 1 mmol of DDQ, and continue stirring for 1 h. The solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.226 g of compound I-32 with a yield of 38%. ESI MS: Theoretical calculation m/z: 496.26, actual test m/z: 496.30. λ ^abs. _max /nm = 706nm, λ ^em _max /nm = 734nm, Ф _f = 0.26.

Example 19

Dissolve 0.510 g (0.001 mmol) of compound I-14 in 5 ml of anhydrous DMF, then add 1 mmol of NaCN, stir at room temperature for 24 h, then add 2 ml of THF solution containing 1 mmol of DDQ, and continue stirring for 1 h. The solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.235 g of compound I-33, with a yield of 44%. ESI MS: theoretical calculation m/z: 436.24, actual test m/z: 436.22. λ ^abs. _max /nm = 705nm, λ ^em _max /nm = 735nm, Ф _f = 0.25.

Example 20

Dissolve 0.500 g (0.001 mmol) of compound I-16 in 5 ml of anhydrous DMF, then add 1 mmol of NaCN, stir at room temperature for 24 h, then add 2 ml of THF solution containing 1 mmol of DDQ, and continue stirring for 1 h. The solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.221 g of compound I-34 with a yield of 42%. ESI MS: theoretical calculation m/z: 426.22, actual test m/z: 426.27. λ ^abs. _max /nm = 694nm, λ ^em _max /nm = 725nm, Ф _f = 0.34.

Example 21

Dissolve 0.501 g (0.001 mmol) of compound I-18 in 5 ml of anhydrous DMF, then add 1 mmol of NaCN, stir at room temperature for 24 h, then add 2 ml of THF solution containing 1 mmol of DDQ, and continue stirring for 1 h. The solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.200 g of compound I-35 with a yield of 38%. ESI MS: Theoretical calculation m/z: 427.21, actual test m/z: 427.19. λ ^abs. _max /nm = 696nm, λ ^em _max /nm = 725nm, Ф _f = 0.37.

Example 22

Dissolve 0.490 g (0.001 mmol) of compound I-21 in 5 ml of anhydrous DMF, then add 1 mmol of NaCN, stir at room temperature for 24 h, then add 2 ml of THF solution containing 1 mmol of DDQ, and continue stirring for 1 h. The solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.185 g of compound I-36, with a yield of 35%. ESI MS: Theoretical calculation m/z: 430.21, actual test m/z: 430.24. λ ^abs. _max /nm = 694nm, λ ^em _max /nm = 720nm, Ф _f = 0.37.

Example 23

Dissolve 0.460 g (0.001 mmol) of compound I-23 in 5 ml of anhydrous DMF, then add 1 mmol of NaCN, stir at room temperature for 24 h, then add 2 ml of THF solution containing 1 mmol of DDQ, and continue stirring for 1 h. The solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.175 g of compound I-37 with a yield of 36%. ESI MS: theoretical calculation m/z: 386.22, actual test m/z: 386.21. λ ^abs. _max /nm = 694nm, λ ^em _max /nm = 720nm, Ф _f = 0.39.

Example 24

Dissolve 0.521 g (0.001 mmol) of compound I-25 in 5 ml of anhydrous DMF, then add 1 mmol of NaCN, stir at room temperature for 24 h, then add 2 ml of THF solution containing 1 mmol of DDQ, and continue stirring for 1 h. The solvent was removed under vacuum, dried, and separated by column chromatography to obtain 0.191 g of compound I-38 with a yield of 35%. ESI MS: Theoretical calculation m/z: 447.19, actual test m/z: 447.17. λ ^abs. _max /nm = 694nm, λ ^em _max /nm = 726nm, Ф _f = 0.35.

Example 25

Compound II-4 was prepared from m-dimethylaminophenol and trifluoroacetic anhydride according to literature (Chem. Eur. J., 2013, 19, 16556-16565), with a yield of 45%.

0.261 g (0.001 mol) of compound II-13 and 0.161 g (0.001 mol) of compound III-1 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 4 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.087 g of compound I-39, with a yield of 18%. ESI MS: theoretical calculation m/z: 387.17, actual test m/z: 387.16. λ ^abs. _max /nm = 674nm, λ ^em _max /nm = 705nm, Ф _f = 0.48.

0.486g (0.001mol) of compound I-39, 0.202g (0.001mol) of di-tert-butyl dicarbonate and 0.101g (0.001mol) of triethylamine were added to 10 ml of anhydrous dichloromethane respectively, After stirring for 24 hours at room temperature, the solvent was removed under vacuum, dried, and purified by column chromatography to obtain 0.516 g of compound I-40 with a yield of 88%. ESI MS: theoretical calculation m/z: 487.22, actual test m/z: 487.25. λ ^abs. _max /nm = 680nm, λ ^em _max /nm = 710nm, Ф _f = 0.45.

Example 26

Compound II-4 was prepared from 8-hydroxyjulolidine and trifluoroacetic anhydride according to literature (Chem. Eur. J., 2013, 19, 16556-16565), with a yield of 52%.

0.285 g (0.001 mol) of compound II-14 and 0.189 g (0.001 mol) of compound III-2 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 4 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.113 g of compound I-41, with a yield of 21%. ESI MS: Theoretical calculation m/z: 439.20, actual test m/z: 439.18. λ ^abs. _max /nm = 686nm, λ ^em _max /nm = 720nm, Ф _f = 0.35.

Example 27

Compound II-4 was prepared from 7-hydroxy-1-methyl-1,2,3,4-tetrahydroquinoline and trifluoroacetic anhydride according to literature (Chem.Eur.J.,2013,19,16556-16565) , yield 51%.

0.259 g (0.001 mol) of compound II-15 and 0.189 g (0.001 mol) of compound III-3 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 4 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.128 g of compound I-42, with a yield of 25%. ESI MS: Theoretical calculation m/z: 413.18, actual test m/z: 413.19. λ ^abs. _max /nm = 695nm, λ ^em _max /nm = 720nm, Ф _f = 0.38.

Example 28

Compound II-4 was synthesized from 7-hydroxyl-1-ethyl-2,2,4-trimethyl-1,2-dihydroquinoline according to literature (Chem.Eur.J.,2013,19,16556-16565) Prepared with trifluoroacetic anhydride in 49% yield.

0.299 g (0.001 mol) of compound II-16 and 0.217 g (0.001 mol) of compound III-4 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 4 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.133 g of compound I-43, with a yield of 23%. ESI MS: theoretical calculation m/z: 481.25, actual test m/z: 481.27. λ ^abs. _max /nm = 705nm, λ ^em _max /nm = 738nm, Ф _f = 0.30.

Example 29

Compound II-4 was prepared from m-dimethylaminophenol and trifluoroacetic anhydride according to literature (Chem. Eur. J., 2013, 19, 16556-16565), with a yield of 44%.

0.233 g (0.001 mol) of compound II-17 and 0.365 g (0.001 mol) of compound III-5 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 4 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.086 g of compound I-44, with a yield of 13%. ESI MS: Theoretical calculation m/z: 563.27, actual test m/z: 563.29. λ ^abs. _max /nm = 686nm, λ ^em _max /nm = 602nm, Ф _f = 0.42.

Example 30

Compound II-4 was synthesized from 7-hydroxyl-1-(4-n-butyric acid ethyl ester)-1,2,3,4-tetrahydroquinone according to literature (Chem.Eur.J.,2013,19,16556-16565) morphine and trifluoroacetic anhydride, yield 41%.

0.359 g (0.001 mol) of compound II-18 and 0.215 g (0.001 mol) of compound III-6 were added to 5 ml of concentrated sulfuric acid, and heated and stirred at 100° C. for 4 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.064 g of compound I-45, with a yield of 10%. ESI MS: theoretical calculation m/z: 539.25, actual test m/z: 539.21. λ ^abs. _max /nm = 694nm, λ ^em _max /nm = 724nm, Ф _f = 0.35.

Example 31

Compound II-4 was prepared from 3-(1-piperidinyl)phenol and trifluoroacetic anhydride according to literature (Chem. Eur. J., 2013, 19, 16556-16565), with a yield of 47%.

0.273 g (0.001 mol) of compound II-19 and 0.241 g (0.001 mol) of compound III-7 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 4 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.104 g of compound I-46, with a yield of 18%. ESI MS: Theoretical calculation m/z: 479.23, actual test m/z: 479.21. λ ^abs. _max /nm = 695nm, λ ^em _max /nm = 724nm, Ф _f = 0.35.

Example 32

Compound II-4 was prepared from 3-(4-morpholinyl)phenol and trifluoroacetic anhydride according to literature (Chem. Eur. J., 2013, 19, 16556-16565), with a yield of 46%.

0.275 g (0.001 mol) of compound II-20 and 0.229 g (0.001 mol) of compound III-8 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 4 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.096 g of compound I-47, with a yield of 17%. ESI MS: theoretical calculation m/z: 469.21, actual test m/z: 469.27. λ ^abs. _max /nm = 682nm, λ ^em _max /nm = 715nm, Ф _f = 0.48.

Example 33

Compound II-4 was prepared from 3-(1-piperazine)phenol and trifluoroacetic anhydride according to literature (Chem. Eur. J., 2013, 19, 16556-16565), with a yield of 35%.

0.274 g (0.001 mol) of compound II-21 and 0.231 g (0.001 mol) of compound III-9 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 4 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.085 g of compound I-48, with a yield of 15%. ESI MS: theoretical calculation m/z: 470.20, actual test m/z: 470.18. λ ^abs. _max /nm = 685nm, λ ^em _max /nm = 715nm, Ф _f = 0.45.

Example 34

Compound II-4 was prepared from m-aminophenol and trifluoroacetic anhydride according to literature (Chem. Eur. J., 2013, 19, 16556-16565), with a yield of 33%.

0.205 g (0.001 mol) of compound II-22 and 0.189 g (0.001 mol) of compound III-3 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 4 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.055 g of compound I-49, with a yield of 12%. ESI MS: Theoretical calculation m/z: 359.14, actual test m/z: 359.18. λ ^abs. _max /nm = 680nm, λ ^em _max /nm = 708nm, Ф _f = 0.50.

0.458g (0.001mol) of compound I-49, 0.202g (0.001mol) of di-tert-butyl dicarbonate and 0.101g (0.001mol) of triethylamine were added to 10 ml of anhydrous dichloromethane respectively, After stirring for 24 hours at room temperature, the solvent was removed under vacuum, dried, and purified by column chromatography to obtain 0.463 g of compound I-50 with a yield of 83%. ESI MS: Theoretical calculation m/z: 459.19, actual test m/z: 459.25. λ ^abs. _max /nm = 684nm, λ ^em _max /nm = 710nm, Ф _f = 0.47.

Example 35

Compound II-4 was prepared from m-ethylaminophenol and trifluoroacetic anhydride according to literature (Chem. Eur. J., 2013, 19, 16556-16565), with a yield of 38%.

0.247 g (0.001 mol) of compound II-23 and 0.217 g (0.001 mol) of compound III-4 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 4 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.074 g of compound I-51, with a yield of 14%. ESI MS: theoretical calculation m/z: 429.21, actual test m/z: 429.27. λ ^abs. _max /nm = 682nm, λ ^em _max /nm = 715nm, Ф _f = 0.48.

Example 36

Compound II-4 was prepared from 4-chloro-3-diethylaminophenol and trifluoroacetic anhydride according to literature (Chem. Eur. J., 2013, 19, 16556-16565), with a yield of 46%.

0.295 g (0.001 mol) of compound II-24 and 0.230 g (0.001 mol) of compound III-10 were added to 5 ml of concentrated sulfuric acid, and then heated and stirred at 100° C. for 4 h. After cooling, the reaction solution was slowly poured into 200 ml of ice water, then 1 ml of perchloric acid (70%) was added dropwise under stirring, and a large amount of distilled water was added, and solids were precipitated after standing, filtered, and vacuum-dried , purified by column chromatography to obtain 0.094 g of compound I-52, with a yield of 16%. ESI MS: Theoretical calculation m/z: 490.19, actual test m/z: 490.21. λ ^abs. _max /nm = 684nm, λ ^em _max /nm = 715nm, Ф _f = 0.42.

Example 37

According to the method of Example 1, by 1 mmol compound II-1 and 1 mmol compound III-2, III-3, III-4, III-6, III-7, III-8, III-9, III -10 reaction, compounds I-53, I-54, I-55, I-56, I-57, I-58, I-59, I-60 can be prepared respectively, and the yields are 83%, 85% respectively , 84%, 87%, 88%, 83%, 81%, 80%.

Example 38

According to the method of Example 1, by 1 mmol compound I-53, I-54, I-55, I-56, I-57, I-58, I-59, I-60 and 1 mmol compound CH ₃ MgI reaction, compounds I-61, I-62, I-63, I-64, I-65, I-66, I-67, I-68 can be prepared respectively, and the yields are 41%, 43% respectively , 44%, 45%, 47%, 43%, 42%, 43%.

Example 39

According to the method of embodiment 1 and embodiment 13, by 1 mmol compound I-53, I-54, I-55, I-56, I-57, I-58, I-59, I-60 respectively with 1 Millimole compound NaCN reaction, can make compound I-69, I-70, I-71, I-72, I-73, I-74, I-75, I-76 respectively, productive rate is respectively 31%, 33%, 34%, 35%, 37%, 35%, 33%, 30%.

Example 40

According to the method of Example 25, by 1 mmol compound II-13 and 1 mmol compound III-2, III-3, III-4, III-6, III-7, III-8, III-9, III -10 reaction, compounds I-77, I-78, I-79, I-80, I-81, I-82, I-83, I-84 can be prepared respectively, and the yields are 11%, 14% respectively , 16%, 17%, 21%, 20%, 15%, 12%.

Example 41

According to the method of Example 1, by 1 mmol compound II-2 and 1 mmol compound III-1, III-3, III-4, III-6, III-7, III-8, III-9, III respectively -10 reaction, compounds I-85, I-86, I-87, I-88, I-89, I-90, I-91, I-92 can be prepared respectively, and the yields are 80%, 86% respectively , 89%, 85%, 88%, 83%, 82%, 81%.

Example 42

According to the method of Example 1, by 1 mmol compound I-85, I-86, I-87, I-88, I-89, I-90, I-91, I-92 and 1 mmol compound CH ₃ MgI reaction, compounds I-93, I-94, I-95, I-96, I-97, I-98, I-99, I-100 can be prepared respectively, and the yields are 38% and 46% respectively , 45%, 43%, 45%, 46%, 41%, 40%.

Example 43

According to the method of embodiment 1 and embodiment 13, by 1 mmol compound I-85, I-86, I-87, I-88, I-89, I-90, I-91, I-92 respectively with 1 Millimole compound NaCN reaction, can make compound I-101, I-102, I-103, I-104, I-105, I-106, I-107, I-108 respectively, productive rate is respectively 28%, 31%, 37%, 36%, 35%, 33%, 35%, 32%.

Example 44

According to the method of Example 25, by 1 mmol compound II-14 and 1 mmol compound III-1, III-3, III-4, III-6, III-7, III-8, III-9, III -10 reaction, compounds I-109, I-110, I-111, I-112, I-113, I-114, I-115, I-116 can be prepared respectively, and the yields are 8% and 18% respectively , 14%, 15%, 17%, 12%, 11%, 9%.

Example 45

According to the method of Example 1, by 1 mmol compound II-15 and 1 mmol compound III-1, III-2, III-4, III-5, III-7, III-8, III-9, III respectively -10 reaction, compounds I-117, I-118, I-119, I-120, I-121, I-122, I-123, I-124 can be prepared respectively, and the yields are 75%, 82% respectively , 83%, 73%, 88%, 83%, 84%, 82%.

Example 46

According to the method of Example 1, by 1 mmol compound I-117, I-118, I-119, I-120, I-121, I-122, I-123, I-124 respectively with 1 mmol compound CH ₃ MgI reaction, compounds I-125, I-126, I-127, I-128, I-129, I-130, I-131, I-132 can be prepared respectively, and the yields are 35% and 41% respectively , 43%, 45%, 44%, 46%, 43%, 41%.

Example 47

According to the method of embodiment 1 and embodiment 13, by 1 mmol compound I-117, I-118, I-119, I-120, I-121, I-122, I-123, I-124 respectively with 1 Millimole compound NaCN reaction, can make compound I-133, I-134, I-135, I-136, I-137, I-138, I-139, I-140 respectively, productive rate is respectively 25%, 30%, 33%, 35%, 37%, 34%, 31%, 33%.

Example 48

According to the method of Example 1, by 1 mmol compound II-15 and 1 mmol compound III-1, III-2, III-4, III-5, III-7, III-8, III-9, III respectively -10 reactions, compounds I-141, I-142, I-143, I-144, I-145, I-146, I-147, and I-148 can be prepared respectively, with yields of 9% and 16% respectively , 13%, 5%, 18%, 14%, 11%, 12%.

Example 49

According to the method of Example 1, by 1 mmol compound II-16 and 1 mmol compound III-1, III-2, III-3, III-5, III-7, III-8, III-9, III respectively -10 reaction, compounds I-149, I-150, I-151, I-152, I-153, I-154, I-155, I-156 can be prepared respectively, and the yields are 75%, 84% respectively , 85%, 87%, 72%, 89%, 85%, 83%.

Example 50

According to the method of Example 1, by 1 mmol compound I-149, I-150, I-151, I-152, I-153, I-154, I-155, I-156 respectively with 1 mmol compound CH ₃ MgI reaction, compounds I-157, I-158, I-159, I-160, I-161, I-162, I-163, I-164 can be prepared respectively, and the yields are 37% and 42% respectively , 44%, 40%, 48%, 45%, 41%, 41%.

Example 51

According to the method of embodiment 1 and embodiment 13, by 1 mmol compound I-149, I-150, I-151, I-152, I-153, I-154, I-155, I-156 respectively with 1 Millimole compound NaCN reaction, can make compound I-165, I-166, I-167, I-168, I-169, I-170, I-171, I-172 respectively, productive rate is respectively 26%, 31%, 33%, 32%, 36%, 35%, 32%, 33%.

Example 52

According to the method of Example 1, by 1 mmol compound II-16 and 1 mmol compound III-1, III-2, III-3, III-5, III-7, III-8, III-9, III respectively -10 reaction, compounds I-173, I-174, I-175, I-176, I-177, I-178, I-179, I-180 can be prepared respectively, and the yields are 8%, 12% respectively , 11%, 7%, 15%, 12%, 13%, 14%.

Example 53

According to the method of Example 1, by 1 mmol compound II-17 and 1 mmol compound III-2, III-3, III-4, III-6, III-7, III-8, III-9, III respectively -11 reaction, compounds I-181, I-182, I-183, I-184, I-185, I-186, I-187, I-188 can be prepared respectively, and the yields are 85%, 87% respectively , 86%, 89%, 89%, 85%, 83%, 80%.

Example 54

According to the method of Example 1, by 1 mmol compound I-181, I-182, I-183, I-184, I-185, I-186, I-187, I-188 and 1 mmol compound CH ₃ MgI reaction, compounds I-189, I-190, I-191, I-192, I-193, I-194, I-195, I-196 can be prepared respectively, and the yields are 39% and 42% respectively , 44%, 45%, 47%, 46%, 42%, 43%.

Example 55

According to the method of embodiment 1 and embodiment 13, by 1 mmol compound I-181, I-182, I-183, I-184, I-185, I-186, I-187, I-188 respectively with 1 Millimole compound NaCN reaction, can make compound I-197, I-198, I-199, I-200, I-201, I-202, I-203, I-204 respectively, productive rate is respectively 29%, 31%, 32%, 36%, 35%, 34%, 31%, 32%.

Example 56

According to the method of Example 1, by 1 mmol compound II-17 and 1 mmol compound III-1, III-2, III-4, III-5, III-7, III-8, III-9, III respectively -10 reaction, compounds I-205, I-206, I-207, I-208, I-209, I-210, I-211, I-212 can be prepared respectively, and the yields are 10%, 12% respectively , 14%, 17%, 20%, 15%, 13%, 12%.

Example 57

According to the method of Example 1, by 1 mmol compound II-18 and 1 mmol compound III-2, III-3, III-4, III-6, III-7, III-8, III-9, III respectively -11 reaction, compounds I-213, I-214, I-215, I-216, I-217, I-218, I-219, I-220 can be prepared respectively, and the yields are 72%, 81% respectively , 79%, 63%, 84%, 81%, 80%, 76%.

Example 58

According to the method of Example 1, by 1 mmol compound I-213, I-214, I-215, I-216, I-217, I-218, I-219, I-220 respectively with 1 mmol compound CH ₃ MgI reaction, compounds I-221, I-222, I-223, I-224, I-225, I-226, I-227, I-228 can be prepared respectively, and the yields are 33% and 38% respectively , 40%, 35%, 44%, 43%, 41%, 42%.

Example 59

According to the method of embodiment 1 and embodiment 13, by 1 mmol compound I-213, I-214, I-215, I-216, I-217, I-218, I-219, I-220 respectively with 1 Millimole compound NaCN reaction, can make compound I-229, I-230, I-231, I-232, I-233, I-234, I-235, I-236 respectively, productive rate is respectively 27%, 32%, 34%, 30%, 36%, 33%, 31%, 30%.

Example 60

According to the method of Example 1, by 1 mmol compound II-18 and 1 mmol compound III-1, III-2, III-4, III-5, III-7, III-8, III-9, III respectively -10 reaction, compounds I-237, I-238, I-239, I-240, I-241, I-242, I-243, I-244 can be prepared respectively, and the yields are 7%, 12% respectively , 13%, 8%, 16%, 12%, 9%, 10%.

Example 61

According to the method of Example 1, by 1 mmol compound II-19 and 1 mmol compound III-2, III-3, III-4, III-6, III-7, III-8, III-9, III respectively -11 reaction, compounds I-245, I-246, I-247, I-248, I-249, I-250, I-251, I-252 can be prepared respectively, and the yields are 81%, 84% respectively , 86%, 73%, 88%, 85%, 84%, 83%.

Example 62

According to the method of Example 1, by 1 mmol compound I-245, I-246, I-247, I-248, I-249, I-250, I-251, I-252 respectively with 1 mmol compound CH ₃ MgI reaction, compounds I-253, I-254, I-255, I-256, I-257, I-258, I-259, I-260 can be prepared respectively, and the yields are 37% and 42% respectively , 43%, 35%, 44%, 43%, 42%, 40%.

Example 63

According to the method of embodiment 1 and embodiment 13, by 1 mmol compound I-245, I-246, I-247, I-248, I-249, I-250, I-251, I-252 respectively with 1 Millimole compound NaCN reaction, can make compound I-261, I-262, I-263, I-264, I-265, I-266, I-267, I-268 respectively, productive rate is respectively 28%, 31%, 33%, 32%, 36%, 34%, 32%, 33%.

Example 64

According to the method of Example 1, by 1 mmol compound II-19 and 1 mmol compound III-1, III-2, III-4, III-5, III-7, III-8, III-9, III respectively -10 reactions, compounds I-269, I-270, I-271, I-272, I-273, I-274, I-275, I-276 can be prepared respectively, and the yields are 11%, 14% respectively , 13%, 9%, 16%, 15%, 13%, 11%.

Example 65

According to the method of Example 1, by 1 mmol compound II-20 and 1 mmol compound III-2, III-3, III-4, III-6, III-7, III-8, III-9, III -11 reaction, compounds I-277, I-278, I-279, I-280, I-281, I-282, I-283, I-284 can be prepared respectively, and the yields are 82%, 84% respectively , 85%, 71%, 86%, 88%, 85%, 82%.

Example 66

According to the method of Example 1, by 1 mmol compound I-277, I-278, I-279, I-280, I-281, I-282, I-283, I-284 and 1 mmol compound CH ₃ MgI reaction, compounds I-285, I-286, I-287, I-288, I-289, I-290, I-291, I-292 can be prepared respectively, and the yields are 36% and 42% respectively , 43%, 34%, 45%, 47%, 42%, 41%.

Example 67

According to the method of embodiment 1 and embodiment 13, by 1 mmol compound I-277, I-278, I-279, I-280, I-281, I-282, I-283, I-284 respectively with 1 Millimole compound NaCN reaction, can make compound I-293, I-294, I-295, I-296, I-297, I-298, I-299, I-300 respectively, and productive rate is respectively 28%, 32%, 33%, 31%, 37%, 38%, 34%, 32%.

Example 68

According to the method of Example 1, by 1 mmol compound II-20 and 1 mmol compound III-1, III-2, III-4, III-5, III-7, III-8, III-9, III -10 reactions, compounds I-301, I-302, I-303, I-304, I-305, I-306, I-307, and I-308 can be prepared respectively, with yields of 9% and 12% respectively , 13%, 8%, 14%, 16%, 13%, 10%.

Example 69

According to the method of Example 1, by 1 mmol compound II-21 and 1 mmol compound III-2, III-3, III-4, III-6, III-7, III-8, III-9, III -11 reaction, compounds I-309, I-310, I-311, I-312, I-313, I-314, I-315, I-316 can be prepared respectively, and the yields are 80% and 82% respectively , 81%, 69%, 84%, 85%, 82%, 78%.

Example 70

According to the method of Example 1, by 1 mmol compound I-309, I-310, I-311, I-312, I-313, I-314, I-315, I-316 respectively with 1 mmol compound CH ₃ MgI reaction, compounds I-317, I-318, I-319, I-320, I-321, I-322, I-323, I-324 can be prepared respectively, and the yields are 33% and 39% respectively , 41%, 32%, 43%, 45%, 40%, 38%.

Example 71

According to the method of embodiment 1 and embodiment 13, by 1 mmol compound I-309, I-310, I-311, I-312, I-313, I-314, I-315, I-316 respectively with 1 Millimole compound NaCN reaction, can make compound I-325, I-326, I-327, I-328, I-329, I-330, I-331, I-332 respectively, productive rate is respectively 27%, 30%, 32%, 31%, 34%, 36%, 32%, 30%.

Example 72

According to the method of Example 1, by 1 mmol compound II-21 and 1 mmol compound III-1, III-2, III-4, III-5, III-7, III-8, III-9, III respectively -10 reaction, compounds I-333, I-334, I-335, I-336, I-337, I-338, I-339, I-340 can be prepared respectively, and the yields are 7%, 11% respectively , 12%, 6%, 14%, 15%, 13%, 11%.

Example 73

According to the method of Example 1, by 1 mmol compound II-22 and 1 mmol compound III-2, III-3, III-4, III-6, III-7, III-8, III-9, III -11 reaction, compounds I-341, I-342, I-343, I-344, I-345, I-346, I-347, I-348 can be prepared respectively, and the yields are 75%, 74% respectively , 69%, 78%, 83%, 81%, 78%, 73%.

Example 74

According to the method of Example 1, by 1 mmol compound I-341, I-342, I-343, I-344, I-345, I-346, I-347, I-348 and 1 mmol compound CH ₃ MgI reaction, compounds I-349, I-350, I-351, I-352, I-353, I-354, I-355, I-356 can be prepared respectively, and the yields are 32% and 41% respectively , 30%, 42%, 44%, 40%, 42%, 33%.

Example 75

According to the method of embodiment 1 and embodiment 13, by 1 mmol compound I-341, I-342, I-343, I-344, I-345, I-346, I-347, I-348 respectively with 1 Millimole compound NaCN reaction, can make compound I-357, I-358, I-359, I-360, I-361, I-362, I-363, I-364 respectively, productive rate is respectively 23%, 28%, 25%, 30%, 33%, 34%, 30%, 28%.

Example 76

According to the method of Example 1, by 1 mmol compound II-22 and 1 mmol compound III-1, III-2, III-4, III-5, III-7, III-8, III-9, III respectively -10 reaction, compounds I-365, I-366, I-367, I-368, I-369, I-370, I-371, I-372 can be prepared respectively, and the yields are 6% and 11% respectively , 5%, 12%, 14%, 11%, 12%, 10%.

Example 77

According to the method of Example 1, by 1 mmol compound II-23 and 1 mmol compound III-2, III-3, III-4, III-6, III-7, III-8, III-9, III respectively -11 reaction, compounds I-373, I-374, I-375, I-376, I-377, I-378, I-379, I-380 can be prepared respectively, and the yields are 81%, 82% respectively , 79%, 85%, 87%, 84%, 82%, 83%.

Example 78

According to the method of Example 1, by 1 mmol compound I-373, I-374, I-375, I-376, I-377, I-378, I-379, I-380 respectively with 1 mmol compound CH ₃ MgI reaction, compounds I-381, I-382, I-383, I-384, I-385, I-386, I-387, I-388 can be prepared respectively, and the yields are 34% and 41% respectively , 33%, 44%, 47%, 45%, 41%, 42%.

Example 79

According to the method of embodiment 1 and embodiment 13, by 1 mmol compound I-373, I-374, I-375, I-376, I-377, I-378, I-379, I-380 respectively with 1 Millimole compound NaCN reaction, can make compound I-389, I-390, I-391, I-392, I-393, I-394, I-395, I-396 respectively, and productive rate is respectively 27%, 31%, 30%, 33%, 35%, 34%, 31%, 32%.

Example 80

According to the method of Example 1, by 1 mmol compound II-23 and 1 mmol compound III-1, III-2, III-4, III-5, III-7, III-8, III-9, III respectively -10 reactions, compounds I-397, I-398, I-399, I-400, I-401, I-402, I-403, and I-404 can be prepared respectively, with yields of 9% and 14% respectively , 13%, 7%, 16%, 15%, 11%, 13%.

Example 81

According to the method of Example 1, by 1 mmol compound II-24 and 1 mmol compound III-1, III-2, III-4, III-5, III-7, III-8, III-9, III respectively -10 reactions, compounds I-405, I-406, I-407, I-408, I-409, I-410, I-411, I-412 can be prepared respectively, and the yields are 81%, 85% respectively , 84%, 72%, 83%, 87%, 82%, 79%.

Example 82

According to the method of Example 1, by 1 mmol compound I-405, I-406, I-407, I-408, I-409, I-410, I-411, I-412 respectively with 1 mmol compound CH ₃ MgI reaction, compounds I-413, I-414, I-415, I-416, I-417, I-418, I-419, I-420 can be prepared respectively, and the yields are 35% and 43% respectively , 42%, 40%, 44%, 46%, 41%, 42%.

Example 83

According to the method of embodiment 1 and embodiment 13, by 1 mmol compound I-405, I-406, I-407, I-408, I-409, I-410, I-411, I-412 respectively with 1 Millimole compound NaCN reaction, can make compound I-421, I-422, I-423, I-424, I-425, I-426, I-427, I-428 respectively, productive rate is respectively 29%, 32%, 33%, 30%, 35%, 37%, 34%, 33%.

Example 84

According to the method of Example 1, by 1 mmol compound II-24 and 1 mmol compound III-1, III-2, III-4, III-5, III-7, III-8, III-9, III respectively -10 reactions, compounds I-429, I-430, I-431, I-432, I-433, I-434, I-435, I-436 can be prepared respectively, and the yields are 10%, 16% respectively , 17%, 11%, 19%, 21%, 15%, 13%.

Example 84

The method for preparing compound I-1 in Example 1 was repeated, with the difference that the heating temperature was changed from 100°C to "60°C", the reaction time was changed to 4h, and the rest of the conditions were kept unchanged to prepare a compound with structural formula I.

According to the preparation method of the above-mentioned Examples 1-84, in the product structure I that can be obtained, R ₁ , R ₂ , R ₆ , and R ₇ can be independently hydrogen, lower hydrocarbon group, ether group, substituted alkyl group or acyl group; R ₃ , R ₄ , R ₅ , R ₈ , R ₉ , and R ₁₀ are independently hydrogen, lower hydrocarbon or halogen; R ₁₁ is hydrogen, methyl, cyano or trifluoromethyl; is an anion;

Wherein, the lower hydrocarbon groups in R ₁ , R ₂ , R ₆ , and R ₇ can be linear, branched or cyclic; the hydrocarbon groups contain 1-6 carbon atoms;

Among them, the number of carbon atoms in the ether group in R ₁ , R ₂ , R ₆ , and R ₇ is 3-6, and the number of oxygen atoms is ≤2;

Among them, the substituted alkyl groups in R ₁ , R ₂ , R ₆ , and R ₇ are linear or branched; Substituting 1-6 carbon atom alkyl group, ω-halogen substituting 1-6 carbon atom alkyl group, ω-hydroxyl substituting 1-6 carbon atom alkyl group, ω-amino group substituting 1-6 carbon atom alkyl group or ω-mercapto substituted 1-6 carbon atom alkyl; wherein, the ω-formate group is 2-5 carbon atom alkyl formate group in the substituted 1-6 carbon atom alkyl ; The carboxamide group in the ω-formamide group substituting the 1-6 carbon atom alkyl group is a 2-5 carbon atom hydrocarbon group formamide group;

Among them, the acyl group in R ₁ , R ₂ , R ₆ , and R ₇ is an alkylacyl group with 2-6 carbon atoms, tert-butoxycarbonyl, benzoyl, benzoyl with 1-6 carbon atoms or halogen-substituted benzoyl Acyl; The R ₁₁ is hydrogen, methyl, cyano or trifluoromethyl;

Wherein halogen is fluorine, chlorine, bromine or iodine.

The obtained near-infrared high quantum yield dyes of the present invention are applied to nucleolar RNA fluorescent labeling experiments:

According to the method in the "Application of Near-infrared Bioluminescent Dyes in Live Cell Imaging" provided in the Chinese invention patent with publication number CN103725758A, the embodiments of the present invention 3, 7, 15, 16, 27, 28, 37, 40 Fluorescent dye compounds I-6, I-15, I-29, I-30, I-42, I-43, I-54, I-78, I-95, I-191, I-255, and I-393 are applied to the fluorescent labeling of nucleolar RNA in A549 cells and Hela cells. The difference is that the final concentration of the dye solution in step 3) is 400-450nM, and the results are shown in Fig. 1-14 shown. It can be seen from Figure 1-14 that under the condition of preparing a lower concentration of dye solution, this series of dyes stains the cytoplasm and also stains the nucleus and inner kernel. In the figure, the cytoplasm has fluorescence, and the nucleolus in the nucleus also has fluorescence. Fluorescence, the entire staining range is large. Moreover, the fluorescence quantum yield of the dye is high, and the dye concentration required for dyeing is also very low, which avoids certain interference of the high concentration dye on the normal physiological functions of living cells and reduces the toxicity of the cells.

Compound dyes I-54 and I-78 obtained by the present invention were digested with DNase and RNase respectively and compared with commercially available DNA and RNA dyes, as shown in Figure 15, the results show that compounds I-54 and I-78 enter the nucleus Stained for nucleolar RNA.

Comparing the cell staining effects of the obtained compound dyes I-54 and I-78 of the present invention with the commercially available RNA dye SYTO RNASelect respectively, as shown in Figure 16, the results show that I-54 and I-78 have better nuclei than SYTO RNASelect Kernel RNA staining, and the damage to the cells is small, reducing the interference of background fluorescence.

Apparently, the above-mentioned embodiments of the present invention are only examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. Those of ordinary skill in the art can also make It is impossible to exhaustively list all the implementation modes here, and any obvious changes or changes derived from the technical solutions of the present invention are still within the scope of protection of the present invention.

Claims (10)

1. a kind of high quantum production rate dyestuff of new near-infrared, it is characterised in that with following structural formula I：

R₁、R₂、R₆、R₇Independently for hydrogen, lower alkyl, ether, substitution alkyl or acyl group；R₃、R₄、R₅、 R₈、R₉、R₁₀Independently for hydrogen, lower alkyl or halogen；R₁₁For hydrogen, methyl, cyano group or trifluoromethyl；For Anion；

The R₁、R₂、R₆、R₇Middle lower alkyl is straight chain, side chain or ring-type；The lower alkyl is former comprising 1-6 carbon Son；

Carbon number is 3-6, oxygen atomicity≤2 in the ether；

The substitution alkyl is straight or branched；The substitution alkyl be ω-formic acid ester group replace 1-6 carbon atom alkyl, ω- Formamido replaces 1-6 carbon atom alkyl, ω-halogen to replace 1-6 carbon atom alkyl, ω-hydroxyl to replace 1-6 carbon atom Alkyl, omega-amino replace 1-6 carbon atom alkyl or ω-sulfydryl to replace 1-6 carbon atom alkyl；Wherein, the ω-formic acid Ester group replaces formic acid ester group in 1-6 carbon atom alkyl to be 2-5 carbon atom alkyl formic acid ester group；Described ω-formamido takes It is 2-5 carbon atom hydrocarbyl radical formamido for formamido in 1-6 carbon atom alkyl；

The acyl group is 2-6 carbon atom alkyl acyl group, tertbutyloxycarbonyl, benzoyl, 1-6 carbon atom substituted benzene formyl Base or halogen-substituted formoxyl；

The halogen is fluorine, chlorine, bromine or iodine；

The R₃、R₄、R₅、R₈、R₉、R₁₀In lower alkyl be methyl or ethyl；

It is describedFor any organic or inorganic anion of balancing charge.

2. dyestuff according to claim 1, it is characterised in that R₁With R₃、R₁With R₁₀、R₂With R₃、R₂With R₁₀、 R₆With R₅、R₆With R₈、R₇With R₅、R₇With R₈、R₁With R₂Or R₆With R₇Following Ia-In structures can be individually formed：

Wherein R is hydrogen, methyl or ethyl；Y is C, O or NR.

3. dyestuff according to claim 1 or 2, it is characterised in that

Described R₁、R₂、R₆、R₇In lower alkyl for methyl, ethyl, propyl group, isopropyl, cyclopropyl, pi-allyl, Butyl, isobutyl group or the tert-butyl group；

The ether is CH₂CH₂OCH₃、CH₂CH₂OCH₂CH₃、CH₂CH₂OCH₂CH₃OH、 CH₂CH₂OCH₂CH₃OCH₃Or CH₂CH₂OCH₂CH₃OCH₂CH₃；

ω-the 2-5 carbon atom alkyl formic acid ester groups replace 1-6 carbon atom alkyl to be (CH₂)_mCOO(CH₂)_nCH₃Or (CH₂)_mCOOC(CH₃)₃, wherein m is 1-4, and n is 0-3；

ω-the 2-5 carbon atom hydrocarbyl radical formamidos replace 1-6 carbon atom alkyl to be (CH₂)_mCONH(CH₂)_nCH₃ Or (CH₂)_mCON[(CH₂)_nCH₃]₂, wherein m is 1-4, and n is 0-3；

The ω-halogen replaces 1-6 carbon atom alkyl to be (CH₂)_mCl、(CH₂)_mBr or (CH₂)_mI, wherein m For 1-6；

The ω-hydroxyl replaces 1-6 carbon atom alkyl to be (CH₂)_mOH, wherein m are 1-6；

The omega-amino replaces 1-6 carbon atom alkyl to be (CH₂)_mNH₂, wherein m is 1-6；

The ω-sulfydryl replaces 1-6 carbon atom alkyl to be (CH₂)_mSH, wherein m are 1-6；

The 2-6 carbon atom alkyl acyl group is acetyl group, propiono, bytyry or tertiary bytyry；

The 1-6 carbon atom substituted benzoyl is methyl substituted benzoyl, ethyl substituted benzoyl, propyl group substituted benzene Formoxyl, butyl substituted benzoyl or tert-butyl group substituted benzoyl；

The halogen-substituted formoxyl is chlorine substituted benzoyl, bromine substituted benzoyl or iodine substituted benzoyl.

4. the preparation method of the high quantum production rate dyestuff of new near-infrared as claimed in claim 1, it is characterised in that structure R in Formulas I₁₁Preparation for hydrogen or the dyestuff of trifluoromethyl comprises the following steps：

1 mM of compound II and 1 mM of compound III are added in the 5-15 milliliters of concentrated sulfuric acids, in a heated condition Reaction 1-4 hours, reaction solution is slowly added dropwise in the frozen water of 20-100 times of volume after cooling, adds 0.01-1 times of volume Mass percentage concentration is 70% perchloric acid, is sufficiently stirred for, and separates out solid, and filtering, vacuum drying, column chromatography for separation must be produced Thing；

The temperature of the heating is 60-100 DEG C；

The structural formula of the compound II is as follows：

The structural formula of the compound III is as follows：

Wherein, R₁、R₂、R₆、R₇Independently for hydrogen, lower alkyl, ether, substitution alkyl or acyl group；R₃、R₄、 R₅、R₈、R₉、R₁₀Independently for hydrogen, lower alkyl or halogen；R₀For hydrogen or trifluoromethyl.

5. the preparation method of the high quantum production rate dyestuff of new near-infrared as claimed in claim 1, it is characterised in that structure R in Formulas I₁₁Preparation for the dyestuff of methyl comprises the following steps：

1) 1 mM of compound II and 1 mM of compound III are added in the 5-15 milliliters of concentrated sulfuric acids, in fire-bar Reacted 1-4 hours under part, reaction solution is slowly added dropwise in the frozen water of 20-100 times of volume after cooling, 0.01-1 times of body is added Long-pending mass percentage concentration is 70% perchloric acid, is sufficiently stirred for, and separates out solid, filtering, vacuum drying, column chromatography for separation Obtain product 1-a；

2) 1 mM of I-a is dissolved in 5 milliliters of anhydrous THF, in N₂It is added dropwise in 3-6 milliliters and contains under protective condition There is 1 mM of CH₃After MgI THF solution, stirring at normal temperature 24h, 2-5 milliliters are added containing 1 mM of DDQ THF solution, continues to stir 1-12h, reaction solution is spin-dried for into rear column chromatography for separation and obtains product；

The temperature of the heating is 60-100 DEG C；

The structural formula of the compound II is as follows：

The structural formula of the compound III is as follows：

Wherein, R₁、R₂、R₆、R₇Independently for hydrogen, lower alkyl, ether, substitution alkyl or acyl group；R₃、R₄、 R₅、R₈、R₉、R₁₀Independently for hydrogen, lower alkyl or halogen；R₀For hydrogen.

6. the preparation method of the high quantum production rate dyestuff of new near-infrared as claimed in claim 1, it is characterised in that structure R in Formulas I₁₁Preparation for the dyestuff of cyano group comprises the following steps：

1. 1 mM of compound II and 1 mM of compound III are added in the 5-15 milliliters of concentrated sulfuric acids, in heating condition Reaction solution, is slowly added dropwise in the frozen water of 20-100 times of volume, adds 0.01-1 times of volume by lower reaction 1-4 hours after cooling Mass percentage concentration be 70% perchloric acid, be sufficiently stirred for, separate out solid, filtering, vacuum drying, column chromatography for separation obtains Product 1-a；

2. 1 mM of obtained I-a is dissolved in 5-20 milliliters of dry DMFs, adds 1 mM of NaCN, normal temperature Stir after 24h, add the THF solution containing 1 mM of DDQ in 2-5 milliliters, reaction solution is spin-dried for rear column chromatography point From obtaining product；

The temperature of the heating is 60-100 DEG C；

The structural formula of the compound II is as follows：

The structural formula of the compound III is as follows：

Wherein, R₁、R₂、R₆、R₇Independently for hydrogen, lower alkyl, ether, substitution alkyl or acyl group；R₃、R₄、 R₅、R₈、R₉、R₁₀Independently for hydrogen, lower alkyl or halogen；R₀For hydrogen.

7. the preparation method according to claim any one of 4-6, it is characterised in that

The R₁、R₂、R₆、R₇Middle lower alkyl is straight chain, side chain or ring-type；The lower alkyl is former comprising 1-6 carbon Son；

Carbon number is 3-6, oxygen atomicity≤2 in the ether；

The substitution alkyl is straight or branched；The substitution alkyl be ω-formic acid ester group replace 1-6 carbon atom alkyl, ω- Formamido replaces 1-6 carbon atom alkyl, ω-halogen to replace 1-6 carbon atom alkyl, ω-hydroxyl to replace 1-6 carbon atom Alkyl, omega-amino replace 1-6 carbon atom alkyl or ω-sulfydryl to replace 1-6 carbon atom alkyl；Wherein, the ω-formic acid Ester group replaces formic acid ester group in 1-6 carbon atom alkyl to be 2-5 carbon atom alkyl formic acid ester group；Described ω-formamido takes It is 2-5 carbon atom hydrocarbyl radical formamido for formamido in 1-6 carbon atom alkyl；

The acyl group is 2-6 carbon atom alkyl acyl group, tertbutyloxycarbonyl, benzoyl, 1-6 carbon atom substituted benzene formyl Base or halogen-substituted formoxyl；

The halogen is fluorine, chlorine, bromine or iodine；

The R₃、R₄、R₅、R₈、R₉、R₁₀In lower alkyl be methyl or ethyl.

8. the preparation method according to claim any one of 4-6, it is characterised in that the compound II and compound III Following IIa-IIg and IIIa-IIIg structures can be individually formed：

Wherein R is hydrogen, methyl or ethyl；Y is C, O or NR.

9. preparation method according to claim 7, it is characterised in that

The R₁、R₂、R₆、R₇In lower alkyl for methyl, ethyl, propyl group, isopropyl, cyclopropyl, pi-allyl, Butyl, isobutyl group or the tert-butyl group；

The ether is CH₂CH₂OCH₃、CH₂CH₂OCH₂CH₃、CH₂CH₂OCH₂CH₃OH、 CH₂CH₂OCH₂CH₃OCH₃Or CH₂CH₂OCH₂CH₃OCH₂CH₃；

ω-the 2-5 carbon atom alkyl formic acid ester groups replace 1-6 carbon atom alkyl to be (CH₂)_mCOO(CH₂)_nCH₃Or (CH₂)_mCOOC(CH₃)₃, wherein m is 1-4, and n is 0-3；

ω-the 2-5 carbon atom hydrocarbyl radical formamidos replace 1-6 carbon atom alkyl to be (CH₂)_mCONH(CH₂)_nCH₃ Or (CH₂)_mCON[(CH₂)_nCH₃]₂, wherein m is 1-4, and n is 0-3；

The ω-halogen replaces 1-6 carbon atom alkyl to be (CH₂)_mCl、(CH₂)_mBr or (CH₂)_mI, wherein m For 1-6；

The ω-hydroxyl replaces 1-6 carbon atom alkyl to be (CH₂)_mOH, wherein m are 1-6；

The omega-amino replaces 1-6 carbon atom alkyl to be (CH₂)_mNH₂, wherein m is 1-6；

The ω-sulfydryl replaces 1-6 carbon atom alkyl to be (CH₂)_mSH, wherein m are 1-6；

The 2-6 carbon atom alkyl acyl group is acetyl group, propiono, bytyry or tertiary bytyry；

The 1-6 carbon atom substituted benzoyl is methyl substituted benzoyl, ethyl substituted benzoyl, propyl group substituted benzene Formoxyl, butyl substituted benzoyl or tert-butyl group substituted benzoyl；

The halogen-substituted formoxyl is chlorine substituted benzoyl, bromine substituted benzoyl or iodine substituted benzoyl.

10. the high quantum production rate dyestuff of the new near-infrared as described in claim any one of 1-3 is used as the nucleolar RNA of near-infrared The application of dyestuff, it is characterised in that the application is in the fluorescence labeling of nucleolar RNA.