CN108948032B

CN108948032B - Synthesis and application of 2, 5-diphenyl-2H-1, 2, 3-triazole-4-carboxylic acid rhodamine B derivative

Info

Publication number: CN108948032B
Application number: CN201811064931.2A
Authority: CN
Inventors: 何文英; 吴禄勇; 李建玲; 刘艳萍; 冯华杰; 丁国华
Original assignee: Hainan Normal University
Current assignee: Hainan Normal University
Priority date: 2018-09-12
Filing date: 2018-09-12
Publication date: 2020-08-25
Anticipated expiration: 2038-09-12
Also published as: CN108948032A

Abstract

The invention relates to a synthesis method and application of 2, 5-diphenyl-2H-1, 2, 3-triazole-4-carboxylic acid rhodamine B derivative, wherein the derivative has a structure shown in formula I：

Description

Synthesis and application of 2, 5-diphenyl-2H-1, 2, 3-triazole-4-carboxylic acid rhodamine B derivative

Technical Field

The invention belongs to the field of organic synthesis and cell imaging, and particularly relates to synthesis and application of a 2, 5-diphenyl-2H-1, 2, 3-triazole-4-carboxylic acid rhodamine B derivative.

Background

With the rapid development of the industry, the environmental pollution poses a great threat to the environment while the economic effect is brought. And heavy metal ion Hg²⁺The pollution of (2) is not negligible. Mercury, one of the most toxic heavy metal ions, is discharged into soil or water, and poses a threat to the human life by enriching the human life through a food chain, so that various diseases caused by mercury, such as alzheimer disease, water deficiency, acral pain and uremia, have serious influence on human life. Fluorescent probes based on 1,2, 3-triazole-rhodamine have been reported, but the application of the probes in cell imaging has not been reported. The invention provides a method for preparing high-sensitivity Hg from rhodamine hydrazide and triazole carboxylate by one step²⁺The probe is recognized by ions and applied to cell imaging.

Disclosure of Invention

The invention provides a 2, 5-diphenyl-2H-1, 2, 3-triazole-4-carboxylic acid rhodamine B derivative with a structure shown in a formula I, which is characterized in that the structure shown in the formula I is as follows:

another embodiment of the present invention provides a process for the preparation of a compound of formula I as described above, characterized by the steps of:

dissolving compound 1 and compound 2 in CH₂Cl₂In Fe (acac)₃And Et₃N to obtain a compound shown in the formula I;

wherein the mol ratio of the compound 1 to the compound 2 is 1:0.8-1.0, Fe (acac)₃Is used in a molar amount of 0.05-0.08 times the amount of compound 1, CH being used per millimole of compound 1₂Cl₂Volume of (2) is 10-15mL, using Et₃The volume of N is 0.4-0.5 mL. The reaction temperature is preferably at room temperature and the reaction time is preferably 20 to 24 hours.

Another embodiment of the present invention provides the use of a compound of formula I in the detection of Hg²⁺The use of (1).

Another embodiment of the present invention provides the use of a compound of formula I for imaging HeLa cells.

Compared with the prior art, the invention has the advantages that: the invention utilizes ferric acetylacetonate (Fe (acac)₃) The one-step synthesis of the 2, 5-diphenyl-2H-1, 2, 3-triazole-4-carboxylic acid rhodamine B derivative is successfully realized by taking the derivative as a catalyst, and the derivative synthesized by the invention has Hg-sensitive activity²⁺The detection sensitivity of the probe is high, and the probe can be successfully used for imaging HeLa cells.

Drawings

FIG. 1 is a drawing of a compound of formula I¹³A CNMR map;

FIG. 2 is a plot of the fluorescence selectivity spectrum of compounds of formula I for each ion;

FIG. 3 shows that the compounds of formula I recognize Hg at different pH²⁺A fluorescence intensity map of the ion of (a);

FIG. 4 is a photograph of fluorescence images of HeLa cells of example 5 of the present invention.

Detailed Description

In order to facilitate a further understanding of the invention, the following examples are provided to illustrate it in more detail. However, these examples are only for better understanding of the present invention and are not intended to limit the scope or the principle of the present invention, and the embodiments of the present invention are not limited to the following.

Example 1

Dissolving compound 1(1.0mmol) and compound 2(1.0mmol) in CH₂Cl₂(15mL), Fe (acac) was added at room temperature₃(0.08mmol) and Et₃N (0.5mL), stirring for reaction for 20 hours, detecting by TLC that the raw material completely disappears, concentrating the reaction solution, and then performing silica gel column chromatography (200-mesh 300-mesh silica gel) with petroleum ether/ethyl acetate (V: 3: 1-4: 1) as an eluent to obtain 619mg of the compound of the formula I with the yield of 87.9%. As shown in fig. 1:¹H NMR(400MHz,CDCl₃):(ppm)＝1.36(t,J＝7.2Hz,12H),3.34(q,J＝7.2Hz,8H),6.37(m,3H),6.77(m,2H),7.32(m,1H),7.35(m,4H),7.45(m,2H),7.53(m,2H),7.52(m,2H),7.84(m,3H),7.87(m,1H)；¹³CNMR(400MHz,CDCl₃)(ppm)＝171.07,164.97,158.32,153.82,151.18,149.44,148.93,138.96,137.32,133.15,129.46,129.34,129.20,129.04,129.01,128.78,128.33,128.25,128.02,124.13,123.51,119.18,107.99,104.44,97.59,77.32,77.00,76.68,66.32,60.31,44.31,29.62,20.96,14.12,12.53.HRMS(ESI)(m/z):[M+H]⁺calcd for C₄₃H₄₂N₇O₃,704.3349；found,704.3348.

example 2

Dissolving compound 1(1.0mmol) and compound 2(1.0mmol) in CH₂Cl₂To (15mL) was added Et at room temperature₃N (0.5mL), after stirring the reaction for 20 hours, TLC showed no new spots (i.e. no reaction) generated in addition to compounds 1 and 2.

Example 3 fluorescence selectivity Spectroscopy of Compounds of formula I

As shown in fig. 2, each ion (1.0 × 10) was assayed in DMF-water (v/v ═ 1:1, Tris-HCl, pH 7.4) solution^-6mol/L) in the presence of a compound of the formula I (2 × 10)^-7mol/L) fluorescence selectivity diagram, in which Hg is irradiated under 562nm exciting light²⁺Upon addition a distinct characteristic emission peak at 577nm appears and a color change is observed.

Example 4 Effect of pH on Compounds of formula I

As shown in FIG. 3, the fluorescence intensity (a) of the compound of formula I in DMF-water solution at various pH conditions was measured; and measuring the addition of Hg at different pH²⁺The change in fluorescence intensity after the reaction (b). The compound of formula I has no fluorescence signal at pH 4.0-10.0, and Hg is added²⁺After that, the fluorescence intensity is at pHThe enhancement was evident at 3.0-8.0 and was strongest at pH 7.4.

Example 5 cellular imaging applications

CO at 37 deg.C₂Experiments were performed by culturing HeLa cells in a cell incubator for 24 hours. As shown in fig. 4, a. bright field imaging of HeLa cells in DMEM culture; B. fluorescence imaging of HeLa cells in DMEM culture; C. bright field imaging of compounds of formula I in HeLa cells; D. fluorescence imaging of compounds of formula I in HeLa cells; E. the compound of formula I is administered to Hg in HeLa cells²⁺Bright field imaging of (1); F. the compound of formula I is administered to Hg in HeLa cells²⁺The concentration of the compound of formula I is 1.0 × 10^-5Hg is added in mol/L²⁺Has a concentration of 2.5 × 10^-5mol/L. The above experimental results indicate that the compounds of formula I of the present invention can be used for cellular imaging.

Claims

1. A preparation method of 2, 5-diphenyl-2H-1, 2, 3-triazole-4-carboxylic acid rhodamine B derivatives with a structure shown in formula I is characterized by comprising the following steps:

the mol ratio of the compound 1 to the compound 2 is 1:0.8-1.0, Fe (acac)₃Is used in a molar amount of 0.05-0.08 times the amount of compound 1, CH being used per millimole of compound 1₂Cl₂Volume of (2) is 10-15mL, using Et₃The volume of N is 0.4-0.5 mL; the reaction temperature is selected from room temperature, and the reaction time is 20-24 hours.