CN111423426A - Asymmetric 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10-five-membered heterocyclic anthracene compound and application thereof - Google Patents

Abstract

The invention belongs to the field of aggregation-induced luminescent materials and cell dyes, and relates to a method for synthesizing an asymmetric 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-five-membered heterocyclic anthracene compound. The purpose is to provide an asymmetric 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-five-membered heterocyclic anthracene fluorescent compound and a preparation method and application thereof. In the compound provided by the invention, 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(thiophen-2-yl)anthracene (BPTA) (that is, X is S in formula I) and 9-(N -tert-Butyloxycarbonyl-2-pyrrolyl)-10-(furan-2-yl)anthracene (BPFA) (that is, X is O in formula I) has significant AIE characteristics, and can stain biological cells, which can As a cell marker; and the compound provided by the present invention has a simple preparation method and low cost.

Description

Asymmetric 9-(N-tert-Butyloxycarbonyl-2-pyrrolyl)-10-Five-membered Heterocyclyl Anthracene Compounds and their applications

technical field

The invention belongs to the field of aggregation-induced luminescent materials and cell dyes, and relates to a synthesis method of an asymmetric 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-five-membered heterocyclic anthracene compound and its application in cell dyeing .

Background technique

Fluorescence is an important luminescence phenomenon that has attracted extensive attention in the past decade. Fluorescent molecules are ideal for imaging, sensing, biological process monitoring, optoelectronic devices, organic light-emitting diodes, and other fields due to their high sensitivity, non-invasiveness, and good spatiotemporal resolution. However, most fluorescent materials are inherently hydrophobic, which hinders their practical applications in the environment, water quality monitoring, and biological systems. Unfortunately, the vast majority of luminescent materials emit in dilute solutions, but show weak or no emission in solid or concentrated solutions, a phenomenon known as aggregation-induced fluorescence quenching (ACQ). This phenomenon has become a major obstacle for the application of organic luminophores, especially in microscopic imaging and biosensing. Encouragingly, some anti-ACQ materials were reported by Tang and Park in 2001 and 2002, respectively, and named them aggregation-induced fluorescence (AIE) and aggregation-induced fluorescence enhanced (AIEE) materials. Such materials exhibit strong fluorescence in concentrated solution and solid state, but do not emit light in dilute solution. Most AIE materials are based on the structural design and synthesis of silole, tetraphenylethylene (TPE), distyryl anthracene (DSA), triphenylamine and cyano-substituted diarylethene derivatives, with an intramolecular motion-limited mechanism. (RIR) as the mechanism by which the AIE phenomenon occurs. As an important class of π-conjugated molecules, anthracene derivatives have unique properties in the construction of organic light-emitting materials, and are also one of the earliest materials developed for use as organic light-emitting diodes. However, like traditional fluorescent materials, anthracene is also affected by the ACQ effect, so it is necessary to design and synthesize anthracene core materials with AIE properties with anthracene as the core.

SUMMARY OF THE INVENTION

In previous work, four symmetrical 9,10-diheterocyclic anthracene (DHA) compounds were designed and synthesized, namely 9,10-dithienylanthracene (DTA), 9,10-difuranthracene (DFA) , 9,10-bis-(N-tert-butyloxycarbonyl-pyridin-2-yl)anthracene (DBPA) and 9,10-bis(1H-pyridin-2-yl)anthracene (DPA). In addition to DBPA, these three DHA derivatives have typical aggregation-induced emission (AIE) properties. We found that the factors affecting the AIE properties of DHA series AIE molecules are the twisted configuration, steric hindrance and packing properties of the heterocycle. Among them, the ACQ phenomenon of DBPA molecule attracted our attention the most. According to the structural characteristics analyzed by the X-ray single crystal diffraction pattern, we found that the reason for its ACQ is the large steric hindrance at the heterocyclic N atom, which makes the twisted configuration of DBPA almost impossible to occur in solution Conformation planarization. However, among the four species, DBPA has the largest fluorescence quantum yield (Φ _f =0.860). We tried to improve the ACQ properties of DBPA by replacing one of the N-tert-butyloxycarbonyl-pyridin-2-yl groups in DBPA, and retain the N-tert-butyloxycarbonyl-pyridin-2-yl group in DBPA for DHA series molecules The effect of fluorescence quantum yield. We designed 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(thiophen-2-yl)anthracene (BPTA) and 9-(N-tert-butyloxycarbonyl-2-pyrrolyl) )-10-(furan-2-yl)anthracene (BPFA) two species. The ultraviolet fluorescence properties of its solid, liquid and single crystal tristates were tested, and its application in cell imaging technology was also tested.

In view of this, the present invention aims to provide an asymmetric 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-five-membered heterocyclic anthracene fluorescent compound and a preparation method and application thereof. Among the compounds provided by the present invention, 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(thiophen-2-yl)anthracene (BPTA) (that is, X in formula I is S) and 9-(N -tert-butyloxycarbonyl-2-pyrrolyl)-10-(furan-2-yl)anthracene (BPFA) (that is, X in formula I is O) has significant AIE properties, and can stain biological cells, which can be As a cell marker; and the compound provided by the present invention has a simple preparation method and low cost.

In order to achieve the above-mentioned purpose of the invention, the present invention provides the following technical solutions:

A 9,10-dipyrrolyl anthracene fluorescent compound, having the structure shown in formula I:

In formula I: X is S, or X is O.

The present invention provides a method for preparing the asymmetric 9,10-diheterocyclyl anthracene (DHA) fluorescent compound described in the above scheme, comprising the following steps:

(i) When X in formula I is S, the compound shown in formula I is 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(thiophen-2-yl)anthracene, and the preparation method includes the following step:

(1) Under protective atmosphere, mix 9,10-dibromoanthracene, 2-boronic acid-N-tert-butyloxycarbonylpyrrole, palladium catalyst, basic compound and solvent to carry out Suzuki coupling reaction to obtain 9-( N-tert-butyloxycarbonyl-2-pyrrolyl)-10-bromoanthracene; the molar ratio of the 9,10-dibromoanthracene and 2-boronic acid-N-tert-butyloxycarbonylpyrrole is 1:1～ 1.3;

(2) Under protective atmosphere, mix 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-bromoanthracene, 2-boronic acid-thiophene, palladium catalyst, basic compound and solvent to carry out Suzuki coupling combined reaction to obtain 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(thiophen-2-yl)anthracene; the 9-(N-tert-butyloxycarbonyl-2-pyrrolyl) The molar ratio of -10-bromoanthracene and 2-boronic acid-thiophene is 1:1～1.5;

(ii) when X is O in formula I, the compound shown in formula I is 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(furan-2-yl)anthracene, and the preparation method includes the following step:

(1) Under protective atmosphere, mix 9,10-dibromoanthracene, 2-boronic acid-N-tert-butyloxycarbonylpyrrole, palladium catalyst, basic compound and solvent to carry out Suzuki coupling reaction to obtain 9-( N-tert-butyloxycarbonyl-2-pyrrolyl)-10-bromoanthracene; the molar ratio of the 9,10-dibromoanthracene and 2-boronic acid-N-tert-butyloxycarbonylpyrrole is 1:1～ 1.3;

(2) Under protective atmosphere, mix 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-bromoanthracene, 2-boronic acid-furan, palladium catalyst, basic compound and solvent to carry out Suzuki coupling combined reaction to obtain 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(furan-2-yl)anthracene; the 9-(N-tert-butyloxycarbonyl-2-pyrrolyl) The molar ratio of -10-bromoanthracene and 2-boronic acid group-furan is 1:1～1.5;

Preferably, the palladium catalysts in (i) and (ii) independently comprise one of Pd(PPh ₃ ) ₄ , PdCl ₂ (dppf) ₂ , Pd(dppf)Cl ₂ and Pd(OAc) ₂ or several.

Preferably, the basic compounds in (i) and (ii) independently include Na ₂ CO ₃ , Ba(OH) ₂ , K ₃ PO ₄ , Cs ₂ CO ₃ , K ₂ CO ₃ , TiOH, KF, One or more of CsF, NaOH and i-PrNEt ₂ .

Preferably, the solvents in (i) and (ii) independently include one or more of THF, CH ₂ Cl ₂ , DMF and CH ₃ CN.

Preferably, the Suzuki coupling reaction in (i) and (ii) is carried out under heating and refluxing conditions, and the reaction time is independently 11-13 h.

The present invention provides the application of the asymmetric 9,10-di-five-membered heterocyclic anthracene fluorescent compound described in the above scheme in cell fluorescence imaging and organic photoluminescence materials.

The beneficial effects of the present invention

(1) The present invention provides two asymmetric 9,10-di-five-membered heterocyclic anthracene fluorescent compounds, which have the structure shown in formula I and have remarkable AIE characteristics, wherein 9-(N-tert-butyloxycarbonyl The fluorescence intensity of -2-pyrrolyl)-10-(thiophen-2-yl)anthracene (BPTA) in the aggregated state was 6.4 times higher than that in the non-aggregated solution, 9-(N-tert-butyloxycarbonyl- The aggregated fluorescence intensity of 2-pyrrolyl)-10-(furan-2-yl)anthracene (BPFA) compound was enhanced 2.4 times in dilute solution.

(2) The present invention provides a method for preparing the asymmetric 9,10-di-five-membered heterocyclic anthracene fluorescent compound described in the above scheme. The preparation method provided by the present invention comprises N-tert-butoxycarbonyl-2-pyrroleboronic acid, Using 2-boronic acid-furan, 2-boronic acid-thiophene and 9,10-dibromoanthracene as the main raw materials, the asymmetric 9,10-two-five-membered heterocycle of the present invention can be obtained through a simple Suzuki coupling reaction The anthracene compound has a simple preparation method. Compared with the common tetraphenylene and distyryl anthracene AIE compounds, the compound provided by the present invention has relatively lower preparation cost, simpler synthesis method and milder preparation conditions, and is more suitable for industrialization. production and application prospects.

(3) The present invention also provides the asymmetric 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-five-membered heterocyclic anthracene fluorescent compound described in the above scheme in cell fluorescence imaging and organic photoluminescence Applications in luminescent materials. All the asymmetric 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-five-membered heterocyclic anthracene fluorescent compounds provided by the present invention can effectively enter HeLa cells. Therefore, the compounds provided by the present invention can be used as cells Fluorescent markers have potential applications in the field of cell dyes.

Description of drawings

Fig. 1 is the fluorescence spectrogram of compound BPTA and the fluorescence photo of solution under the condition of different moisture volume percentage;

Fig. 2 is the fluorescence spectrogram of compound and BPFA and the fluorescence photo of solution under different moisture volume percentage conditions;

Fig. 3 is the fluorescence emission spectra of compounds BPTA and BPFA in different organic solvents;

Fig. 4 is the fluorescence emission spectra of compounds BPTA and BPFA in solid state;

Fig. 5 is the ultraviolet absorption spectrogram of compound (A) BPTA and (B) BPFA under different solvents;

Fig. 6 is the ultraviolet absorption spectra of compounds BPTA and BPFA in DMF;

Fig. 7 is the ultraviolet absorption spectrum of compound BPTA and BPFA in solid state;

Figure 8 is a particle size diagram of compounds (A) BPTA and (B) BPFA when aggregated at a volume ratio of DMF:H ₂ O=1:9;

Fig. 9 is the scanning electron microscope photograph of compound (A) BPTA and BPFA in aggregated state;

Figure 10 is a crystal ORTEP diagram of compounds (A) BPTA and (B) BPFA;

Figure 11 is a crystal unit cell packing diagram of compounds (A) BPTA and (B) BPFA;

Fig. 12 is the fluorescence imaging image of compound BPTA and BPFA cells.

Detailed ways

The present invention provides an asymmetric 9,10-two-five-membered heterocyclic anthracene fluorescent compound, which has the structure shown in formula I:

In formula I: X is S, or X is O.

In the present invention, when X in formula I is S, the compound represented by formula I is 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(thiophen-2-yl)anthracene (BPTA) , the structural formula is shown in formula II:

In the present invention, when X in formula I is O, the compound represented by formula I is 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(furan-2-yl)anthracene (BPFA) , the structural formula is shown in formula III:

All of the asymmetric 9,10-di-five-membered heterocyclic anthracene fluorescent compounds provided by the present invention have significant aggregation-induced luminescence enhancement properties, can stain biological cells, and can be used as cell markers. In addition, the asymmetric 9,10-two-five-membered heterocyclic anthracene fluorescent compound provided by the present invention has a simple molecular structure, is easy to synthesize, has low cost of raw materials, and is easy to modify, and can be continuously optimized for design to improve its aggregation-induced luminescence properties and cell staining. It has good application prospects in the fields of organic optoelectronic materials, biochemical detection and cell fluorescence imaging.

The present invention provides the preparation method of the asymmetric 9,10-di5-membered heterocyclic anthracene aggregation-induced luminescent compound described in the above scheme, comprising the following steps:

(i) when X in formula I is S, the compound represented by formula I is 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(thiophen-2-yl)anthracene (BPTA), prepared The method includes the following steps:

(1) Under protective atmosphere, mix 9,10-dibromoanthracene, 2-boronic acid-N-tert-butyloxycarbonylpyrrole, palladium catalyst, basic compound and solvent to carry out Suzuki coupling reaction to obtain 9-( N-tert-butyloxycarbonyl-2-pyrrolyl)-10-bromoanthracene; the molar ratio of the 9,10-dibromoanthracene and 2-boronic acid-N-tert-butyloxycarbonylpyrrole is 1:1～ 1.5.

(2) Under protective atmosphere, mix 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-bromoanthracene, 2-thiopheneboronic acid, palladium catalyst, basic compound and solvent to carry out Suzuki coupling reaction , to obtain 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(thiophen-2-yl)anthracene; the 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10 - The molar ratio of bromoanthracene and 2-thiopheneboronic acid is 1:1 to 1.5.

In the present invention, the reaction formula of the Suzuki coupling reaction in (i) is shown in formula V;

In the present invention, the protective atmosphere is preferably argon; the palladium catalyst preferably includes one of Pd(PPh ₃ ) ₄ , PdCl ₂ (dppf) ₂ , Pd(dppf)Cl ₂ and Pd(OAc) ₂ or several; the basic compounds preferably include Na ₂ CO ₃ , Ba(OH) ₂ , K ₃ PO ₄ , Cs ₂ CO ₃ , K ₂ CO ₃ , TiOH, KF, CsF, NaOH and i-PrNEt ₂ ( One or more of diethylisopropylamine); the solvent preferably includes one or more of THF, CH ₂ Cl ₂ , DMF and CH ₃ CN.

In the present invention, the molar concentration of the palladium catalyst in the reaction solution is preferably 10%; the addition ratio of the 9,10-dibromoanthracene and the solvent is preferably 10mmol～0.1mol: 80～250mL; more preferably 50mmol～80mmol: 100～200mL; the basic compound is preferably added in the form of an aqueous solution, and the normality of the basic compound solution is preferably 5equiv.; the addition of the 9,10-dibromoanthracene and the basic compound solution The amount ratio is preferably 10 mmol to 0.1 mol:60 to 120 mL.

In the present invention, the Suzuki coupling reaction in (i) is preferably carried out under heating and refluxing conditions, and the reaction time is preferably 11-13 hours, more preferably 12 hours.

In a specific embodiment of the present invention, it is preferable to add 9,10-dibromoanthracene and palladium catalyst into the solvent first, stir for 30 min, and then add 2-boronic acid-N-tert-butyloxycarbonylpyrrole and basic compound solution, Then heat to reflux.

After the Suzuki coupling reaction is completed, the present invention preferably carries out post-processing to the reaction solution, and the post-processing preferably comprises the following steps:

The Suzuki coupling reaction solution was cooled to room temperature and the solvent was removed by rotary evaporation to obtain a rotary evaporation residue;

The rotary evaporation residue is extracted, and the obtained organic phase is successively dried and filtered to obtain a filtrate;

The filtrate was rotary evaporated to remove the solvent and then subjected to column chromatography to obtain pure 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(thiophen-2-yl)anthracene.

In the present invention, the extraction agent for extraction is preferably dichloromethane; the drying agent for drying is preferably anhydrous magnesium sulfate; the eluent for column chromatography is preferably a petroleum ether-dichloromethane mixture; The volume ratio of petroleum ether and dichloromethane in the mixed solution is preferably 1:1; the present invention has no special requirements for the specific methods of operations such as rotary evaporation and filtration, and methods well known to those skilled in the art can be used. In the present invention, the obtained 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-bromoanthracene obtained after column chromatography is a pale yellow solid.

(ii) when X is O in formula I, the compound shown in formula I is 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(furan-2-yl)anthracene (BPFA), to prepare The method includes the following steps:

(1) Under protective atmosphere, mix 9,10-dibromoanthracene, 2-boronic acid-N-tert-butyloxycarbonylpyrrole, palladium catalyst, basic compound and solvent to carry out Suzuki coupling reaction to obtain 9-( N-tert-butyloxycarbonyl-2-pyrrolyl)-10-bromoanthracene; the molar ratio of the 9,10-dibromoanthracene and 2-boronic acid-N-tert-butyloxycarbonylpyrrole is 1:1～ 1.5.

(2) Under protective atmosphere, mix 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-bromoanthracene, 2-furanboronic acid, palladium catalyst, basic compound and solvent to carry out Suzuki coupling reaction , to obtain 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(furan-2-yl)anthracene; the 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10 - The molar ratio of bromoanthracene and 2-furanboronic acid is 1:1 to 1.5.

In the present invention, the reaction formula of Suzuki coupling reaction in described (i) is shown in formula VI;

In the present invention, the protective atmosphere is preferably argon; the palladium catalyst preferably includes one of Pd(PPh ₃ ) ₄ , PdCl ₂ (dppf) ₂ , Pd(dppf)Cl ₂ and Pd(OAc) ₂ or several; the basic compounds preferably include Na ₂ CO ₃ , Ba(OH) ₂ , K ₃ PO ₄ , Cs ₂ CO ₃ , K ₂ CO ₃ , TiOH, KF, CsF, NaOH and i-PrNEt ₂ ( One or more of diethylisopropylamine); the solvent preferably includes one or more of THF, CH ₂ Cl ₂ , DMF and CH ₃ CN.

In the present invention, the molar concentration of the palladium catalyst in the reaction solution is preferably 10%; the addition ratio of the 9,10-dibromoanthracene and the solvent is preferably 10mmol～0.1mol: 80～250mL; more preferably 50mmol～80mmol: 100～200mL; the basic compound is preferably added in the form of an aqueous solution, and the normality of the basic compound solution is preferably 5equiv.; the addition of the 9,10-dibromoanthracene and the basic compound solution The amount ratio is preferably 10 mmol to 0.1 mol:60 to 120 mL.

In the present invention, the Suzuki coupling reaction in (i) is preferably carried out under heating and refluxing conditions, and the reaction time is preferably 11-13 hours, more preferably 12 hours.

In a specific embodiment of the present invention, it is preferable to add 9,10-dibromoanthracene and palladium catalyst into the solvent first, stir for 30 min, and then add 2-boronic acid-N-tert-butyloxycarbonylpyrrole and basic compound solution, Then heat to reflux.

After the Suzuki coupling reaction is completed, the present invention preferably carries out post-processing to the reaction solution, and the post-processing preferably includes the following steps:

The Suzuki coupling reaction solution was cooled to room temperature and the solvent was removed by rotary evaporation to obtain a rotary evaporation residue;

The rotary evaporation residue is extracted, and the obtained organic phase is successively dried and filtered to obtain a filtrate;

The filtrate was rotary evaporated to remove the solvent and then subjected to column chromatography to obtain pure 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(furan-2-yl)anthracene.

In the present invention, the extraction agent for extraction is preferably dichloromethane; the drying agent for drying is preferably anhydrous magnesium sulfate; the eluent for column chromatography is preferably a petroleum ether-dichloromethane mixture; The volume ratio of petroleum ether and dichloromethane in the mixed solution is preferably 1:1; the present invention has no special requirements for the specific methods of operations such as rotary evaporation and filtration, and methods well known to those skilled in the art can be used. In the present invention, the 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(furan-2-yl)anthracene obtained after separation by column chromatography is a pale yellow solid.

The preparation method of the above-mentioned asymmetric 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-five-membered heterocyclic anthracene aggregation-induced luminescent compound provided by the present invention only involves Suzuki coupling reaction and deprotection group reaction , and compared with the common tetrastyrene and distyryl anthracene AIE compounds, the cost of raw materials is low, the synthesis method is simpler, and the preparation conditions are milder, suitable for industrial production, and has broad application prospects.

The present invention also provides the asymmetric 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-five-membered heterocyclic anthracene aggregation-induced luminescent compound described in the above scheme in cell fluorescence imaging and organic photoluminescence Applications in luminescent materials. The asymmetric 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-five-membered heterocyclic anthracene aggregation-induced luminescent compounds provided by the present invention can all effectively enter HeLa cells. Therefore, the compounds provided by the present invention can be used as cell fluorescent markers and have potential application value in the field of cell dyes. And the compound provided by the present invention has high chemical stability and thermal stability, and can be used for preparing organic photoluminescent materials, especially blue light materials.

The solutions provided by the present invention will be described in detail below in conjunction with the examples, but they should not be construed as limiting the protection scope of the present invention.

Example 1

Preparation of 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(thiophen-2-yl)anthracene (BPTA):

(1) Preparation of 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-bromoanthracene:

Under argon protection, 9,10-dibromoanthracene (3.36 g, 10 mmol) and Pd(PPh ₃ ) ₄ (10 mol%) were dissolved in 80.0 mL of THF, stirred for 30 min, and then added 2-boronic acid-N-tertiary Butyloxycarbonylpyrrole (2.10 g, 10.0 mmol) and 60 mL of Na ₂ CO ₃ solution with a concentration of 5.0 equiv, heated to reflux for 12 h, stopped the reaction, and cooled to room temperature. The solvent was evaporated by rotary evaporation, extracted with dichloromethane, and the organic phases were combined and dried over anhydrous magnesium sulfate. Suction filtration, the filtrate was rotary evaporated to remove the solvent, and separated by column chromatography with petroleum ether:dichloromethane=2:1 as the eluent to obtain a white solid (2.80 g, 6.6 mmol), yield: 66%.

(2) 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(thiophen-2-yl)anthracene (BPTA)

Under argon, 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-bromoanthracene (2.1 g, 5.0 mmol) and Pd(PPh ₃ ) ₄ (0.2 g) were dissolved in 30.0 mL In THF, stir for 30 min, add 2-thiopheneboronic acid (0.9 g, 7.5 mmol) and 10 mL of 2.0 mol/L Na ₂ CO ₃ solution, heat to reflux for 12 h, stop the reaction, and cool to room temperature. The solvent was evaporated by rotary evaporation, extracted with dichloromethane, and the organic phases were combined and dried over anhydrous magnesium sulfate. Filtration with suction, rotary evaporation of the filtrate to remove the solvent, and separation by column chromatography with petroleum ether:dichloromethane=2:1 as the eluent to obtain a light yellow solid (1.8 g, 4.3 mmol), yield: 85%.

Structure identification: ¹ H NMR (400MHz, CDCl ₃ , ppm): δ0.53(s, 9H, Boc-CH ₃ ), 6.35(t, 1H), 6.44(t, 1H), 7.10(s, 1H), 7.24(s,1H),7.31(m,4H,J＝8.0Hz),7.54(d,1H,J＝8.0Hz), 7.62(m,3H,J＝8.0Hz),7.77(m,2H,J =8.0Hz); ¹³ C NMR (100 MHz, CDCl ₃ , TMS) δ=25.68, 81.81, 109.98, 110.09, 115.54, 115.63, 120.99, 121.08, 124.55, 121.49, 125.61, 126.15, 126.34, 138.5. 148.32

Example 2

Preparation of 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(furan-2-yl)anthracene (BPFA):

1. 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-bromoanthracene

Under argon protection, 9,10-dibromoanthracene (3.8 g, 11.3 mmol) and Pd(PPh ₃ ) ₄ (0.5 g) were dissolved in 80.0 mL of THF, stirred for 30 min, and then added 2-boronic acid-N-tertiary Butyloxycarbonylpyrrole (2.1 g, 10.0 mmol) and 60 mL of Na ₂ CO ₃ solution with a concentration of 2.0 mol/L were heated to reflux for 12 h to stop the reaction and cooled to room temperature. The solvent was evaporated by rotary evaporation, extracted with dichloromethane, and the organic phases were combined and dried over anhydrous magnesium sulfate. Suction filtration, the filtrate was rotary evaporated to remove the solvent, and separated by column chromatography with petroleum ether:dichloromethane=2:1 as the eluent to obtain a white solid (2.8 g, 6.6 mmol), yield: 66%.

Structure identification: ¹ H NMR (400MHz, CDCl ₃ , ppm): δ 0.51 (s, 9H, Boc-CH ₃ ), 6.30 (s, 1H, pyrrole-H), 6.43 (s, 1H, anthracene-H) ,7.35(t,2H,anthracene-H,J＝8.0Hz),7.51(t,2H,anthracene-H,J＝8.0Hz),7.61(t,2H,anthracene-H,J＝8.0Hz),8.49 (d,2H,pyrrole–H,J＝8.0Hz)

2. 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-(furan-2-yl)anthracene (BPFA)

Under argon, 9-(N-tert-butyloxycarbonyl-2-pyrrolyl)-10-bromoanthracene (2.1 g, 5.0 mmol) and Pd(PPh ₃ ) ₄ (0.2 g) were dissolved in 30.0 mL After stirring for 30 min in THF, 2-furanboronic acid (0.8 g, 7.5 mmol) and 10 mL of 2.0 mol/L Na ₂ CO ₃ solution were added, heated to reflux for 12 h, the reaction was stopped, and cooled to room temperature. The solvent was evaporated by rotary evaporation, extracted with dichloromethane, and the organic phases were combined and dried over anhydrous magnesium sulfate. Suction filtration, the filtrate was rotary evaporated to remove the solvent, and separated by column chromatography with petroleum ether:dichloromethane=2:1 as the eluent to obtain a light yellow solid (1.6 g, 4.3 mmol), yield: 81%.

Structure identification: ¹ H NMR (400MHz, CDCl ₃ , ppm): δ0.52(s, 9H, Boc-CH ₃ ), 6.33(s, 1H), 6.44(s, 1H), 6.60(s, 1H), 6.66 (s, 1H), 7.33 (t, 4H, J=8.0Hz), 7.63 (t, 2H, J=8.0Hz), 7.69 (s, 2H), 8.83 (d, 2H, J=8.0Hz); ¹³ C NMR (100MHz, CDCl ₃ , TMS) δ=25.46, 81.96, 109.73, 111.18, 115.80, 115.63, 120.91, 124.48, 124.71, 125.13, 125.54, 128.00, 130.31, 130.54;

Characterization data:

Figure 1 shows the fluorescence spectrum of the compound BPTA and the fluorescence photos of the solution under different water volume percentage conditions; the concentration of BPTA is 2.0×10 ^-5 mol/L, and the organic solvent used is DMF.

It can be seen from Figure 1 that BPTA is in a dilute solution state when there are many good solvents (DMF), and there is obvious fluorescence at this time. When the volume fraction of the poor solvent (water) in the solution exceeds 80%, BPTA aggregates in water. When nanoparticles are formed, the fluorescence intensity increases significantly, and the fluorescence intensity of the solution in the aggregated state is 6.4 times higher than that in the non-aggregated solution, indicating that the compound BPTA provided by the present invention has significant aggregation-induced emission enhancement (AIEE) properties.

Figure 2 shows the fluorescence spectra of the compound and BPFA and the fluorescence photos of the solution under different water volume percentage conditions; the concentration of BPFA is 2.0×10 ^-5 mol/L, and the organic solvent used is DMF.

It can be seen from Figure 2 that BPFA is in a dilute solution state when there are many good solvents (DMF), and there is obvious fluorescence emission at this time. When the volume fraction of the poor solvent (water) in the solution reaches 90%, BPFA aggregates in water. , forming nanoparticles, the fluorescence intensity of the solution in the aggregated state is 2.4 times higher than that in the non-aggregated solution, indicating that the compound BPFA provided by the present invention has remarkable aggregation-induced emission enhancement (AIEE) properties.

Figure 3 is the fluorescence emission spectrum of compounds BPTA and BPFA in different organic solvents, wherein (A) is the fluorescence emission spectrum of compound BPTA, (B) is the fluorescence emission spectrum of compound BPFA; the concentrations of compounds BPTA and BPFA are both It is 2.0×10 ^-5 mol/L. It can be seen from Figure 3 that the compounds BPTA and BPFA have obvious fluorescence emission in different solvents.

Figure 4 shows the fluorescence emission spectra of compounds BPTA and BPFA in solid state.

It can be seen from Figure 4 that the maximum emission peak positions of the two compounds in the solid state are close, and different five-membered heterocycles have little effect on the fluorescence properties of the two compounds.

Figure 5 shows the UV absorption spectra of compounds BPTA and BPFA in different solvents, in which (A) is the UV absorption spectrum of the compound BPTA, (B) is the UV absorption spectrum of the compound BPFA; the concentrations of the compounds BPTA and BPFA are both 2.0×10 ^-5 mol/L.

Figure 6 is the ultraviolet absorption spectra of compounds BPTA and BPFA in DMF, wherein the concentrations of the compounds BPTA and BPFA are both 2.0×10 ^-5 mol/L.

It can be seen from Figures 5-6 that with the increase of solvent polarity, the position of the maximum UV absorption peaks of the three compounds changes little, indicating that the solution polarity has little effect on the UV absorption performance of the compounds of the present invention.

Fig. 7 is the ultraviolet absorption spectrum of compound BPTA and BPFA in solid state;

Figure 8 is the particle size diagram of compound BPTA and BPFA when aggregated at the volume ratio of DMF:H ₂ O=1:9; wherein (A) is the particle size diagram of compound BPTA, (B) is the particle size diagram of compound BPFA.

It can be seen from Figure 8 that the average particle size of BPTA in the aggregated state is 673 nm, while the average particle size of BPFA in the aggregated state is 710 nm.

Fig. 9 is the scanning electron microscope photo of compound BPTA and BPFA in the aggregated state, wherein (A) is the scanning electron microscope photo of the compound BPTA in the aggregated state, (B) is the scanning electron microscope photo of BPFA in the aggregated state; the test method is: yes Three compound solutions with a concentration of 2.0×10 ^-5 mol/L and a solvent of DMF:H ₂ O=1:9 (volume ratio) were dropped on the conductive insulating adhesive, dried and plated with gold, and scanned with Zessi-SEM electron microscope observation;

It can be further shown from FIG. 9 that in the solution of DMF:H ₂ O=1:9 (volume ratio), all three compounds aggregated.

Figure 10 is the crystal ORTEP diagram of compound BPTA and BPFA, wherein (A) is the crystal ORTEP diagram of compound BPTA, (B) is the crystal ORTEP diagram of BPFA;

Figure 11 is the crystal unit cell packing diagram of compound BPTA and BPFA, wherein (A) is the crystal unit cell packing diagram of compound BPTA (b axis), (B) is the crystal unit cell packing diagram of BPFA (b axis). Cultivation method: Single crystals of BPTA and BPFA were prepared by slow vaporization of DCM and n-hexane mixed solvent at room temperature; test method: their crystal structures were determined by X-ray single crystal diffraction.

Cell imaging experiments:

HeLa cells were cultured in bovine serum supplemented with 10% (mass fraction) DMEM at a culture temperature of 37°C and a culture atmosphere of 5% CO ₂ -95% air. Cells were placed in 20mm cell culture dishes and allowed to persist overnight before experiments. After washing the HeLa cells with phosphate buffered saline (PBS), the substances that stain the cells (ie, BPTA and BPFA, both at a concentration of 20 μM) were incubated in the culture medium for 30 minutes. After washing the HeLa cells with PBS three times, the cells were imaged with an OLYMPUS FV1000 confocal laser scanning microscope, and the fluorescence emission at 460-560 nm was used as the collection channel. The results are shown in Figure 12.

Figure 12 is the fluorescence imaging images of compound BPTA and BPFA cells, wherein A ₁ is the fluorescent dark field imaging image of compound BPTA, B ₁ is the bright field imaging image of compound BPTA, C ₁ is the superimposed field imaging image of compound BPTA; A ₂ is Fluorescence dark-field imaging of compound BPFA, B2 is the bright _- field imaging of compound BPFA, _C2 is the superimposed field imaging of compound BPFA.

It can be seen from Figure 12 that both compounds provided by the present invention can effectively enter HeLa cells. Therefore, the compounds provided by the present invention can be used as cell fluorescent markers and have potential application value in the field of cell dyes.

The above are only the preferred embodiments of the present invention. It should be pointed out that for those skilled in the art, without departing from the principles of the present invention, several improvements and modifications can be made. It should be regarded as the protection scope of the present invention.

Claims (7)

1. An asymmetric 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10-pentanary heterocyclylanthracene compound, characterized in that: has a structure shown in formula I:

in formula I: x is S, or X is O.

2. The asymmetric 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10-penta-heterocyclic anthracene compound of claim 1, which is synthesized by:

(i) when X in the formula I is S, the compound shown in the formula I is 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10- (thiophene-2-yl) anthracene, and the preparation method comprises the following steps:

(1) mixing 9, 10-dibromoanthracene, 2-boric acid group-N-tert-butyloxycarbonyl pyrrole, a palladium catalyst, an alkaline compound and a solvent under the protection of nitrogen to perform Suzuki coupling reaction to obtain 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10-bromoanthracene; the molar ratio of the 9, 10-dibromoanthracene to the 2-boranyl-N-tert-butyloxycarbonyl pyrrole is 1: 1-1.3;

(2) mixing 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10-bromoanthracene, 2-boranyl-thiophene, a palladium catalyst, an alkaline compound and a solvent under the protection of nitrogen to carry out Suzuki coupling reaction to obtain 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10- (thiophene-2-yl) anthracene; the molar ratio of the 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10-bromoanthracene to the 2-boranyl-thiophene is 1: 1-1.5;

(ii) when X in the formula I is O, the compound shown in the formula I is 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10- (furan-2-yl) anthracene, and the preparation method comprises the following steps:

(1) mixing 9, 10-dibromoanthracene, 2-boric acid group-N-tert-butyloxycarbonyl pyrrole, a palladium catalyst, an alkaline compound and a solvent under the protection of nitrogen to perform Suzuki coupling reaction to obtain 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10-bromoanthracene; the molar ratio of the 9, 10-dibromoanthracene to the 2-boranyl-N-tert-butyloxycarbonyl pyrrole is 1: 1-1.3;

(2) mixing 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10-bromoanthracene, 2-boranyl-furan, a palladium catalyst, an alkaline compound and a solvent under the protection of nitrogen to carry out Suzuki coupling reaction to obtain 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10- (furan-2-yl) anthracene; the molar ratio of the 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10-bromoanthracene to the 2-boranyl-furan is 1: 1-1.5.

3. The method for synthesizing the asymmetric 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10-penta-heterocyclic anthracene compound according to claim 2, wherein the method comprises the following steps: the palladium catalyst in the steps (i) and (ii) is Pd (PPh)₃)₄、PdCl₂(dppf)₂、Pd(dppf)Cl₂、Pd(OAc)₂One or more of them are mixed.

4. The method for synthesizing the asymmetric 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10-penta-heterocyclic anthracene compound according to claim 2, wherein the method comprises the following steps: the alkaline compound in (i) and (ii) is Na₂CO₃、Ba(OH)₂、K₃PO₄、Cs₂CO₃、K₂CO₃TiOH, KF, CsF, NaOH and i-PrNEt₂One or more of them are mixed.

5. The method for synthesizing the asymmetric 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10-penta-heterocyclic anthracene compound according to claim 2, wherein the method comprises the following steps: the solvent in (i) and (ii) is THF or CH₂Cl₂DMF and CH₃One or more of CN.

6. The method for synthesizing the asymmetric 9- (N-tert-butyloxycarbonyl-2-pyrrolyl) -10-five-membered heterocyclic anthracene compound according to claim 2, wherein the method comprises the following steps: the Suzuki coupling reaction in the (i) and the (ii) is carried out under the condition of heating reflux, and the reaction time is independently 11-13 h.

7. The use of the method of claim 1 for the synthesis of 9- (N-t-butyloxycarbonyl-2-pyrrolyl) -10-pentanary heterocyclylanthracene compounds for cellular fluorescence imaging and organic photoluminescent materials.

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