Novel BODIPY-based near-infrared open type fluorescent probe for detecting hypochlorous acid as well as preparation method and application of novel BODIPY-based near-infrared open type fluorescent probe

Abstract

The invention discloses a novel BODIPY-based near-infrared open-type fluorescent probe for detecting hypochlorous acid as well as a preparation method and application of the novel BODIPY-based near-infrared open-type fluorescent probe. The fluorescent probe is (5, 5-difluoro-1, 3, 9-trimethyl-7-(4-sulfur morpholinyl styryl)-5H-dipyrrolo-diazaborolo-penta [10] alkene-10-yl)-4-phenol, which is abbreviated as BOD-Y. The invention also discloses a preparation method of the fluorescent probe. The probe BOD-Y can specifically recognize hypochlorous acid, the color of a probe BOD-Y solution added with hypochlorous acid is rapidly changed from green to blue under sunlight, and the fluorescence is also changed from colorless to strong red fluorescence. The detection limit of the probe on hypochlorous acid is as low as 21nM, the probe shows a good linear relation within 0-35 [mu] M, and the response time is within 13 seconds. The probe shows good selectivity on hypochlorous acid, and is not influenced and interfered by other anions, cations, biological mercaptan, active oxygen and the like.

Description

Technical Field

[0001] The invention belongs to the technical field of fluorescent probes and relates to a novel BODIPY-based near-infrared open-type fluorescent probe for detecting hypochlorous acid, a preparation method thereof and an application thereof. Background Art

[0002] Reactive oxygen species, as essential signaling molecules in cells, play an important role in human physiological activities. Hypochlorous acid is one of the most important reactive oxygen species in biological systems, mainly catalyzed by myeloperoxidase under physiological conditions. - It is produced by reacting with H2O2. However, excessive hypochlorous acid can cause oxidation of some biomolecules such as proteins, nucleic acids, lipids, and enzymes, inducing a range of diseases such as cardiovascular disease, neuronal degeneration, arthritis, cancer, and acute coronary syndrome. Due to its weak acidity and strong oxidizing properties, hypochlorous acid is widely used in fabric bleaching, water and environmental disinfection, and food processing. Therefore, developing a highly selective and sensitive method for real-time monitoring of hypochlorous acid in biological and environmental systems is a research task of great significance.

[0003] In recent years, a variety of methods have been developed for detecting hypochlorous acid, such as the starch-potassium iodide method, electroanalysis, chemiluminescence, and chromatography. Among them, fluorescent probes have been widely used in chemical analysis and biological monitoring due to their significant advantages, such as good selectivity and high sensitivity. Although a variety of fluorescent probes capable of analyzing hypochlorous acid fluorescence imaging have been reported, most of these probes have some shortcomings, such as wavelengths mostly in the visible light region, short response times, and poor water solubility, which will affect the application of the probes in biological systems. Therefore, it is very important to design a near-infrared open-type fluorescent probe for detecting hypochlorous acid. Summary of the Invention

[0004] In view of the shortcomings of the prior art, the technical problem to be solved by the present invention is to provide a new BODIPY-based near-infrared open-type fluorescent probe for detecting hypochlorous acid. The fluorescent probe has the advantages of long wavelength (in the near-infrared region), high sensitivity and high selectivity for detecting hypochlorous acid.

[0005] In order to solve the above technical problems, the technical solution adopted by the present invention is:

[0006] A novel BODIPY-based near-infrared open-type fluorescent probe for detecting hypochlorous acid, characterized in that the probe is (5,5-difluoro-1,3,9-trimethyl-7-(4-thiomorpholinylphenyl)-5H-dipyrrolodiazaborol

[10] ene-10-yl)-4-phenol, abbreviated as BOD-Y, with a molecular formula of: C 30 H 30 BF2N3OS, the structural formula is:

[0007]

[0008] The synthesis route of the novel BODIPY-based near-infrared open-type fluorescent probe for detecting hypochlorous acid is as follows:

[0009]

[0010] The specific steps include:

[0011] Dissolve 0.2-0.5 mmol of the compound BOD-OH, 0.8-1.0 mmol of 4-thiomorpholinecarboxaldehyde, and 2-5 mmol of p-toluenesulfonic acid in 10 mL of anhydrous toluene. Raise the temperature to 70°C and gradually add 0.5-1.5 mL of piperidine. Reflux the reaction mixture at 110°C for 5-7 hours (TLC monitoring). Wash the reaction mixture with water, dry it, and distill it. The resulting residue is purified on a silica gel column (petroleum ether:ethyl acetate = 5:1, v / v) to obtain a purple solid, the probe BOD-Y.

[0012] The BOD-Y probe selectively reacts with hypochlorous acid. When hypochlorous acid is added to a solution of the probe, the color of the solution rapidly changes from green to blue, and the fluorescence changes from colorless to red under sunlight. The linear range for hypochlorous acid is 0-35 μM, with a detection limit as low as 21 nM and a response time within 10 seconds.

[0013] Beneficial effects: Compared with the prior art, the BODIPY-based near-infrared open-type fluorescent probe BOD-Y of the present invention changes the color of the probe solution rapidly from green to blue and the fluorescence color from colorless to red in the presence of hypochlorous acid. The intensity of the fluorescence emission spectrum increases with the increase in the concentration of hypochlorous acid and has a significant linear correlation, which can be used for high-sensitivity detection of hypochlorous acid. At the same time, the fluorescent probe responds quickly to hypochlorous acid. In addition, the fluorescent probe shows good selectivity for hypochlorous acid and is not affected by other interfering substances such as active oxygen, active thiols and anions. As a fluorescent probe for detecting hypochlorous acid, BOD-Y has the advantages of convenient synthesis, rapid response, high sensitivity, good selectivity, and large emission wavelength. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is the fluorescence spectrum of the fluorescence intensity at 598-830nm after BOD-Y reacts with different concentrations of hypochlorous acid;

[0015] Figure 2 This is a linear relationship diagram of the fluorescence intensity change after BOD-Y reacts with different concentrations of hypochlorous acid;

[0016] Figure 3It is the fluorescence spectrum of the change of fluorescence intensity after BOD-Y interacts with other test substances (other test substances include blank, Cu 2+ , NO 3- , Na + , HS - , Fe 2+ , Fe 3+ , Cl - , CO3 2- , Br - , K + , I - , SO4 2- , SO3 2- ,HCO3 - , Mg 2+ NO2 - , Zn 2+ , ACO - , H2PO4 - ,SCN - ,Cys,Hcy,GSH,NO,H2O2,*OH,O 2- , ONOO - , 1 O2,) DETAILED DESCRIPTION

[0017] The present invention will be further described below with reference to specific embodiments.

[0018] Example 1

[0019] Preparation of fluorescent probe BOD-Y:

[0020] Compound BOD-OH (0.43 mmol, 0.195 g), 4-thiomorpholinecarboxaldehyde (0.9 mmol, 0.186 g), and p-toluenesulfonic acid (5 mmol, 0.8 g) were dissolved in 50 mL of anhydrous toluene and added to a three-necked flask. The temperature was then raised to 70°C, and piperidine (1 mL) was gradually added. The reaction system was refluxed and stirred at 110°C for 5-7 hours. After the reaction, the reaction mixture was rotary evaporated, washed, and dried. The crude product was purified by silica gel column chromatography (PE:EA = 5:1) to obtain the target compound BOD-Y as a purple solid (35% yield). 1 H NMR (600MHz, DMSO-d6) δ9.81 (s, 1H), 7.46 (s, 3H), 7.27 (s, 1H), 7.13 (s, 2H), 6.99 (s, 2H), 6.8 9(s, 3H), 6.14(s, 1H), 3.71(s, 4H), 2.66(s, 4H), 2.46(s, 3H), 1.48(s, 3H), 1.43(s, 3H); [M+H] +calcd for C 30 H 30 BF2N3OS, 529.22; found, 529.2178.

[0021] Example 2

[0022] BOD-Y was made into 1.5×10 -5 M PBS buffer solution (containing 30% DMF), 10 M sodium hypochlorite solution was diluted with distilled water to concentrations of 0, 0.4×10 -5 , 1.8×10 -5 , 3.8×10 -5 , 6×10 -5 , 8.8×10 -5 , 11.5×10 -5 、14.5×10 -5 , 18×10 -5 , 22×10 -5 , 26×10 -5 M, 30×10 -5 M, 35×10 -5 M solution, add different concentrations of hypochlorous acid solution to the probe solution. Fluorescence emission spectrum of BOD-Y in the presence of different concentrations of hypochlorous acid was measured on a fluorescence spectrophotometer using fluorescence spectrum titration method, with an excitation wavelength of 560nm and collection of fluorescence emission peaks at 598nm-830nm. Figure 1 It can be seen that with the addition of hypochlorous acid, the fluorescence intensity of BOD-Y continues to increase. Then, a linear fit is performed on the relationship between the peak fluorescence intensity of the probe (fluorescence intensity at 694 nm) and the concentration change of hypochlorous acid, and the following equation is obtained: Figure 2 The linear relationship shown (y = 21.8x + 234.5, R 2 =0.983), and further calculation (LOD = 3Sb / m) yielded a detection limit of 21×10 -9 M. Furthermore, upon exposure to hypochlorous acid, the solution's color changes from green to blue. Therefore, the probe can be used as a fluorescent switch-on probe for sensitive detection of changes in hypochlorous acid concentration.

[0023] Example 3

[0024] BOD-Y was prepared to 1.5×10 -5 M solution, and different active oxygen, active sulfur and other anions were dissolved in the aqueous solution to prepare a concentration of 1×10 -2 The fluorescence emission intensity of the probe at 694 nm in the presence of different active oxygen, active sulfur, metal ions and anions was measured using a fluorescence spectrophotometer using a fluorescence spectrophotometer. Figure 3 As shown in Figure 2, only the addition of hypochlorous acid can significantly increase the fluorescence intensity of the probe within 10s, while other analytes (Cu 2+ , NO 3- , Na + , HS - , Fe 2+ , Fe 3+ , Cl - , CO3 2- , Br - , K + , I - , SO4 2- , SO3 2- ,HCO3 - ,Mg 2+ , NO2 - ,Zn 2+ , ACO - , H2PO4 - , SCN - ,Cys,Hcy,GSH,NO,H2O2,·OH,O 2- ,ONOO - , 1 Compared with the reference of hypochlorous acid (HO2, HClO), the fluorescence intensity ratio of the probe did not change significantly. This shows that the compound can be used as a fluorescent probe for the specific and sensitive detection of hypochlorous acid.

Claims

1. A novel BODIPY-based near-infrared open-type fluorescent probe for detecting hypochlorous acid, characterized in that: Its molecular formula is C 30 H 30 BF2N3OS, the structural formula is:

2. The preparation method of a novel BODIPY-based near-infrared open-type fluorescent probe for detecting hypochlorous acid according to claim 1, characterized in that 4-(5,5-difluoro-1,3,7,9-tetramethyl-5H-diazaborolin-10-yl)phenol (referred to as compound BOD-OH) reacts with 4-thiomorpholine carboxaldehyde to obtain the probe (5,5-difluoro-1,3,9-trimethyl-7-(4-thiomorpholinylphenyl)-5H-dipyrrolodiazaborolin-10-yl)-4-phenol, referred to as BOD-Y.

3. The method for preparing a novel BODIPY-based near-infrared open-type fluorescent probe for detecting hypochlorous acid according to claim 2, characterized in that: The specific preparation method includes: Dissolve 0.3-0.5 mmol of the compound BOD-OH, 0.8-1.0 mmol of 4-thiomorpholinecarboxaldehyde, and 2-5 mmol of p-toluenesulfonic acid in 10 mL of anhydrous toluene. Raise the temperature to 70°C and gradually add 0.5-1.5 mL of piperidine. Reflux the reaction mixture at 110°C for 5-7 hours (TLC monitoring). Wash the reaction mixture with water, dry it, and distill it. Purify the residue on a silica gel column (petroleum ether:ethyl acetate = 5:1, v / v) to obtain the purple solid compound, probe BOD-Y.

4. Use of the novel BODIPY-based near-infrared open-type fluorescent probe BOD-Y for detecting hypochlorous acid according to claim 1 in detecting hypochlorous acid.

5. The use according to claim 4, characterized in that The BOD-Y probe selectively reacts with hypochlorous acid. When hypochlorous acid is added to a solution of the probe, the color of the solution rapidly changes from green to blue, and the fluorescence changes from colorless to red under sunlight. The linear range for hypochlorous acid is 0-35 μM, with a detection limit as low as 21 nM and a response time within 10 seconds.

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