阅读说明：本技术 一种哌啶基bodipy类红光荧光染料及其制备方法和应用 (Piperidyl BODIPY red-light fluorescent dye and preparation method and application thereof ) 是由 徐海军 蒋浩 杨剑 田蒋为 徐徐 于 2019-11-04 设计创作，主要内容包括：本发明公开了一种哌啶基BODIPY类红光荧光染料及制备方法和应用,该荧光染料通过1,3,5,7-四甲基-8-(2,4,6-三甲基苯基)氟硼二吡咯化合物与对氟苯甲醛的缩合反应制备。该染料能用于检测溶液pH值,对pH的检测范围在1-7。且该染料具有较好的细胞渗透性,能用于细胞内荧光成像。可作为性能优异的红光染料,在荧光成像、生物荧光分析、荧光传感器、荧光标记等领域具有广泛应用。(The invention discloses a piperidyl BODIPY red-light fluorescent dye and a preparation method and application thereof, wherein the fluorescent dye is prepared by condensation reaction of a 1, 3, 5, 7-tetramethyl-8- (2, 4, 6-trimethylphenyl) BODIPY compound and p-fluorobenzaldehyde. The dye can be used for detecting the pH value of a solution, and the detection range of the pH value is 1-7. And the dye has better cell permeability and can be used for intracellular fluorescence imaging. Can be used as a red dye with excellent performance and has wide application in the fields of fluorescence imaging, biological fluorescence analysis, fluorescence sensors, fluorescence labeling and the like.)

1. The piperidyl BODIPY red fluorescent dye (I) is characterized by having the following structural formula:

2. the preparation method of the piperidyl BODIPY red fluorescent dye (I) as described in claim 1, which is characterized by comprising the following steps: under the anhydrous condition, adding 1, 3, 5, 7-tetramethyl-8- (2, 4, 6-trimethylphenyl) BODIPY and p-fluorobenzaldehyde into dry toluene according to the molar ratio of 1: 0.8-1.0, adding p-toluenesulfonic acid and piperidine, stirring and heating, controlling the temperature at 120-130 ℃ and the reaction time at 16-18 hours, finishing the reaction, extracting, drying, separating and purifying by a silica gel chromatographic column to obtain the piperidyl BODIPY red-light fluorescent dye (I), wherein the reaction formula in the process is as follows:

3. the preparation method of the piperidinyl BODIPY red-light fluorescent dye according to claim 2, wherein the volume ratio of toluene to 1, 3, 5, 7-tetramethyl-8- (2, 4, 6-trimethylphenyl) BODIPY is 15-20 mL: 1 mmol.

4. The preparation method of the piperidinyl BODIPY red fluorescent dye according to claim 2, wherein the molar ratio of 1, 3, 5, 7-tetramethyl-8- (2, 4, 6-trimethylphenyl) BODIPY to p-toluenesulfonic acid is 2: 0.8-1.

5. The application of the piperidyl BODIPY red fluorescent dye as described in claim 1, wherein the piperidyl BODIPY red fluorescent dye has a fluorescence intensity effect on pH in a solution by adopting a fluorescence method, and can be used for pH detection and analysis in the solution, and the detection range is 1-7 at pH.

6. The use of the piperidinyl BODIPY-type red fluorescent dye of claim 1 as a red fluorescent dye in tumor cell imaging.

Technical Field

The invention relates to a fluorescent compound and the field of preparation method and application thereof, in particular to a piperidyl BODIPY red fluorescent dye and the preparation method and the application thereof.

Background

BODIPY (BODIPY) fluorescent substances are a new class of dyes, and have the advantages of small Stokes shift, good stability, insensitivity to acid and alkali, high fluorescence quantum yield, narrow fluorescence excitation and emission peaks, high solubility in a plurality of organic solvents and the like. Most of the absorption and emission wavelengths of the common BODIPY derivatives are about 500nm, so that the application range of the BODIPY derivatives is limited. The development of an efficient and convenient organic synthesis method for carrying out structural modification on BODIPY, and the preparation of BODIPY fluorescent compounds with absorption and emission wavelengths in red light or near infrared regions is a recent research hotspot.

The modified BODIPY derivative is widely applied to the fields of ion probes, molecular probes, fluorescent labeling, analytical chemistry, biological analysis and detection, environmental science and the like. The pH value is an important parameter, and the determination of the pH value has important significance in the aspects of environmental analysis, chemical reaction process control, biomedicine and the like. The physicochemical properties of substances, the reaction performance of the substances in aqueous solution and a plurality of physiological processes in a living system are closely related to the pH value of the environment in which the substances are positioned. Currently, the pH measurement methods mainly include an indicator method, a pH paper method, a glass electrode method, an optical sensor method, and the like. In recent years, monitoring pH changes by using changes in various fluorescence parameters (e.g., fluorescence intensity, fluorescence lifetime, wavelength of fluorescence, etc.) of fluorescent molecules has attracted much interest. The pH probe assay method established based on the change of optical signals has drawn much attention due to its advantages of high sensitivity, good selectivity, and short reaction time, as compared to other pH assay methods.

Cells play an important role in growth of a living body and the like as important life units, and changes in the tissue architecture of cells are closely related to the phenotype of a disease. Therefore, the method has important significance for researches such as cell fluorescence imaging and targeted cell positioning. This not only allows one to gain insight into genomic information, but also provides pathologists with early diagnosis and prognosis of lesion information. However, the existing fluorescent dyes capable of specifically locating cells have the problems of few types, high price, poor repairability and the like. Therefore, the research of cell dyes has been long paid attention by researchers as a hotspot and difficulty in the frontier field. In view of the important significance of cell fluorescence imaging in the fields of cell positioning, cell targeting drug loading, tumor cell photodynamic killing and the like, the synthesized novel organic dye molecule with fluorescence property has high scientific research value and application value in the field of cell fluorescence imaging.

Disclosure of Invention

The purpose of the invention is as follows: aiming at the defects in the prior art, the invention aims to provide a piperidine BODIPY red fluorescent dye and a preparation method and application thereof.

The technical scheme is as follows: in order to achieve the purpose of the invention, the invention adopts the technical scheme that:

the invention relates to a piperidyl BODIPY red fluorescent dye and a preparation method and application thereof, which are characterized in that the fluorescent probe has the following structural formula (I):

a preparation method of a piperidyl-containing BODIPY near-red fluorescent dye comprises the following steps:

1) adding 1, 3, 5, 7-tetramethyl-8- (2, 4, 6-trimethylphenyl) boron dipyrrole, p-fluorobenzaldehyde and p-toluenesulfonic acid into a round bottom flask provided with a Dean-stark device, then dissolving the mixture into 25-30 mL of toluene and 2-4 mL of piperidine, heating the reaction mixture to 120-130 ℃ and refluxing for 16-18 hours. Wherein the mol ratio of the 1, 3, 5, 7-tetramethyl-8- (2, 4, 6-trimethylphenyl) boron dipyrrole to the p-fluorobenzaldehyde is 1: 0.8-1.0.

2) Cooling the reaction mixture to room temperature, extracting with dichloromethane, washing with water, drying the organic layer, distilling under reduced pressure to remove the organic solvent, separating the residue by silica gel column chromatography, and eluting with dichloromethane-petroleum ether to obtain a black-green solid product.

The specific chemical reaction formula is as follows:

in the step (1), the volume ratio of the added 1, 3, 5, 7-tetramethyl-8- (2, 4, 6-trimethylphenyl) BODIPY to the toluene is 1 mmol: 15-20 mL.

In the step (1), the molar ratio of the 1, 3, 5, 7-tetramethyl-8- (2, 4, 6-trimethylphenyl) boron dipyrrole to the p-toluenesulfonic acid is 2: 0.8-1.

In the step (2), the eluent for silica gel column chromatography is petroleum ether and dichloromethane which are 3: 7.

The application of the piperidyl BODIPY red fluorescent dye (I) as a fluorescent probe in the aspect of pH value detection.

The application of the piperidyl BODIPY red fluorescent dye (I) as a red dye with long wavelength in the aspect of tumor cell imaging.

The invention has the advantages of

The invention has the following advantages: the structure, synthesis, fluorescence properties and application of the piperidyl BODIPY red-light fluorescent dye (I) are not reported in the prior art, the performance of the dye is excellent, red fluorescence is emitted in pure organic solvents such as dichloromethane and the like, and the maximum emission wavelength is 671 nm; and as the pH value is reduced, the emission wavelength is blue-shifted, the fluorescence is enhanced, and the probe can be used as a pH fluorescent probe; can also be used as organic fluorescent dye for tumor cell fluorescence imaging; the piperidyl BODIPY red-light fluorescent dye has extremely important application prospect in the fields of living cell imaging, fluorescence sensors, biological fluorescence analysis, fluorescence labeling and the like.

Drawings

FIG. 1 shows the piperidinyl BODIPY type red fluorescent dye (I) in dichloromethane (concentration 1X 10)^-5mol/L) fluorescence spectrum;

FIG. 2 shows the piperidinyl BODIPY-like red fluorescent dyes (I) (concentration 1X 10) at different pH values^-5mol/L) fluorescence spectrum;

FIG. 3 is a laser confocal fluorescence micrograph of piperidinyl BODIPY red-light fluorescent dye (I) incubated with hepatoma carcinoma cell HepG 2;

FIG. 4 is an electrospray high-resolution mass spectrum of the piperidinyl BODIPY red fluorescent dye (I).

Detailed Description

The present invention will be further described with reference to specific examples, but the present invention is not limited to the following examples. Example 1 preparation of a piperidinyl BODIPY-like Red fluorescent dye (I)

1, 3, 5, 7-tetramethyl-8- (2, 4, 6-trimethylphenyl) fluoro is added into a three-neck round-bottom flask under anhydrous conditionBoron dipyrromethene (2mmol, 732mg), p-fluorobenzaldehyde (2.0mmol, 248mg), p-toluenesulfonic acid (0.87mmol, 150mg) were dissolved with 30ml of toluene and 2ml of piperidine. The reaction mixture was heated to 125 ℃ under reflux for 16 hours. Cooling to room temperature, washing with saturated salt water for 2 times, extracting with dichloromethane for 3 times, and spin-drying to obtain crude product. Separation was performed using a silica gel column, and the eluent was dichloromethane and petroleum ether 7: 3, to obtain a dark green solidified compound (I) with a yield of 31%. Nuclear magnetic resonance hydrogen spectrum of compound (I):¹H-NMR(600MHz，CDCl₃，ppm)：δ7.53-7.48(m，4H)，7.19(d，J＝16.2Hz，1H)，6.95(s，2H)，6.88(d，J＝8.7Hz，2H)，6.58(s，1H)，5.96(s，1H)，3.27(t，J＝10.8Hz，3H)，2.59(s，3H)，2.34(s，3H)，2.11(s，6H)，1.72-1.68(m，4H)，1.64-1.54(m，2H)，1.43(s，3H)，1.39(s，3H).HRMS(ESI)：[M+H]⁺，C₃₄H₃₉BF₂N₃calculated：538.3205；found：538.3215.

the synthesis route of the piperidyl BODIPY red fluorescent dye (I) is as follows: