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

Abstract

The invention discloses a novel BODIPY-based near-infrared fluorescent probe for continuously detecting hydrogen sulfide and hypochlorous acid as well as a preparation method and application of the novel BODIPY-based near-infrared fluorescent probe. The fluorescent probe is 4-(3, 7-bis (2-(10-butyl-10H-phenothiazin-3-yl) vinyl)-5, 5-difluoro-1, 9-dimethyl-5H-4, 5-dipyrrolo [1, 3, 2] dinitroboron heterocyclic hexene-10-yl) phenyl 3, 5-dinitrobenzene sulfonate, which is called B-PHS-OP for short. The probe B-PHS-OP can specifically and continuously recognize hydrogen sulfide and hypochlorous acid, the color of a probe B-PHS-OP solution added with hydrogen sulfide is rapidly changed from light blue to green under sunlight, and the fluorescence color of the solution is changed from colorless to dark red under irradiation of 365nm ultraviolet light. Then hypochlorous acid is added, the solution is rapidly changed from green to cyan under sunlight, and the fluorescence color of the solution is rapidly changed from dark red to bright red under irradiation of 365nm ultraviolet light. Therefore, the compound can be used as a fluorescent probe for continuously detecting hydrogen sulfide and hypochlorous acid.

Description

Technical Field

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

[0002] Reactive sulfur and reactive oxygen species, as important intracellular signaling molecules, play crucial roles in the human body. Hydrogen sulfide (HS) is present in the human body and has diverse functions in numerous physiological and pathological processes. The physiological concentration of HS in mammalian blood ranges from 30 to 100 mM, while levels in the brain range from 50 to 160 mM. Abnormal HS concentrations can have various effects on biological processes, leading to numerous diseases such as Alzheimer's disease, diabetes, and Down syndrome. Hypochlorous acid (HOCl) is produced by the myeloperoxidase-catalyzed reaction of chloride ions and hydrogen peroxide in white blood cells under certain physiological conditions and is one of the most important reactive oxygen species in biological systems. In human physiological processes, HOCl protects the immune system from pathogens and bacteria. However, excessive HOCl can lead to the oxidation of biomolecules such as proteins, nucleic acids, lipids, and enzymes, leading to tissue damage, inflammation, and a range of diseases such as cardiovascular disease, kidney disease, and tumors. Therefore, developing efficient and continuous methods for the detection of HS and HOCl is crucial for a better understanding of their biological functions in living systems.

[0003] Currently, the main methods for detecting hydrogen sulfide and hypochlorous acid include colorimetry, iodine titration, and electrochemical methods. Compared to these detection methods, fluorescent probes have been widely used in fields such as life sciences, environmental science, and food science due to their advantages such as rapid detection, high sensitivity, good selectivity, and simple operation. Compared with traditional detection methods, fluorescent probes offer unique advantages such as simple synthesis, excellent sensitivity, high selectivity, and good biocompatibility, and have developed into powerful measurement tools. Although a variety of fluorescent probes capable of fluorescence imaging analysis of hydrogen sulfide and hypochlorous acid have been reported, most of these fluorescent probes are designed for separate detection of hydrogen sulfide and hypochlorous acid, and most of them emit only a single emission signal in the visible light region, which limits their application in biological systems. Therefore, the design of a near-infrared fluorescent probe that can continuously detect hydrogen sulfide and hypochlorous acid is particularly important. 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 BODIPY-based near-infrared fluorescent probe that can continuously detect hydrogen sulfide and hypochlorous acid. The fluorescent probe has a long wavelength (in the near-infrared region), high sensitivity and high selectivity for continuous detection of hydrogen sulfide and 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 fluorescent probe for continuous detection of hydrogen sulfide and hypochlorous acid is 4-(3,7-bis(2-(10-butyl-10H-phenothiazin-3-yl)vinyl)-5,5-difluoro-1,9-dimethyl-5H-4,5-dipyrrolo[1,3,2]diazaborin-10-yl)phenyl 3,5-dinitrobenzenesulfonate, abbreviated as B-PHS-OP, with a molecular formula of C 59 H 51 BF2N6O7S3, the structural formula is:

[0007]

[0008] The method for preparing a novel BODIPY-based near-infrared fluorescent probe for continuous detection of hydrogen sulfide and hypochlorous acid uses 4-(3,7-bis(2-(10-butyl-10H-phenothiazine-3-yl)vinyl)-5,5-difluoro-1,9-dimethyl-5H-4,5-dipyrrolo[1,3,2]diazaborohexin-10-yl)phenol (B-PHS-OH for short) as a raw material, and undergoes a substitution reaction with 2,4-dinitrobenzenesulfonyl chloride to obtain the probe 4-(3,7-bis(2-(10-butyl-10H-phenothiazine-3-yl)vinyl)-5,5-difluoro-1,9-dimethyl-5H-4,5-dipyrrolo[1,3,2]diazaborohexin-10-yl)phenyl 3,5-dinitrobenzenesulfonate, B-PHS-OP for short. The method specifically comprises the following steps:

[0009] (1) Dissolve 15 mmol of B-PHS-OH and 5-10 mL of triethylamine in anhydrous dichloromethane. Then add 20-60 mmol of 2,4-dinitrobenzenesulfonyl chloride to the mixture and stir at room temperature for 6 h. Monitor the reaction by TLC until the starting material spot disappears, then stop the reaction.

[0010] (2) 300-400 mL of dichloromethane was added to the reaction mixture, and then washed with distilled water. The organic phase was dried and distilled, and the resulting crude product was purified by silica gel column (dichloromethane: methanol = 150:1, v / v) to obtain compound B-PHS-OP as a brown solid.

[0011] The probe B-PHS-OP is able to react specifically and continuously with hydrogen sulfide and hypochlorous acid. When hydrogen sulfide is added to a solution of probe B-PHS-OP, the color of the solution rapidly changes from light blue to green in sunlight, and under 365nm ultraviolet light, the fluorescence color of the solution changes from colorless to dark red. Subsequently, when hypochlorous acid is added, the solution rapidly changes from green to cyan in sunlight, and under 365nm ultraviolet light, the fluorescence color of the solution also rapidly changes from dark red to intense red. This compound can specifically and continuously identify the content of hydrogen sulfide and hypochlorous acid, with detection limits as low as 89nM and 12nM, respectively.

[0012] Beneficial Effects: Compared with existing technologies, the BODIPY-based near-infrared fluorescent probe B-PHS-OP of the present invention can specifically and continuously detect hydrogen sulfide and hypochlorous acid. Furthermore, the fluorescent probe can sensitively and continuously detect the content of hydrogen sulfide and hypochlorous acid in solutions. As a fluorescent probe for the continuous detection of hydrogen sulfide and hypochlorous acid, B-PHS-OP offers numerous advantages, including ease of synthesis, rapid response, high sensitivity, good selectivity, and a wide emission wavelength, and possesses promising application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 This is the fluorescence spectrum of B-PHS-OP after reacting with different concentrations of hydrogen sulfide;

[0014] Figure 2 This is the fluorescence spectrum of B-PHS-OP after it reacted with 45 μM hydrogen sulfide and then with different concentrations of hypochlorous acid;

[0015] Figure 3 The fluorescence spectra of B-PHS-OP after reacting with hydrogen sulfide and other interfering analytes;

[0016] Figure 4 This is the fluorescence spectrum of B-PHS-OP after reacting with 45μM hydrogen sulfide and then with hypochlorous acid and other interfering analytes. DETAILED DESCRIPTION

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

[0018] Example 1

[0019] The preparation of the novel BODIPY-based near-infrared fluorescent probe for continuous detection of hydrogen sulfide and hypochlorous acid is as follows:

[0020]

[0021] The specific steps are as follows:

[0022] Preparation of B-PHS-OP:

[0023] B-PHS-OH (10 mmol, 8.8 g) and 5 mL of triethylamine were dissolved in 100 mL of anhydrous dichloromethane. 2,4-Dinitrobenzenesulfonyl chloride (10 mmol, 5.3 g) was then slowly added to the mixture at 0°C and stirred for 0.5 h, followed by 6 h at room temperature. After the reaction, the mixture was extracted with water and dichloromethane, washed, and dried over anhydrous sodium sulfate. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 150:1) to obtain a brown solid, the target probe B-PHS-OP, in a 35% yield. 1 H NMR (600MHz, DMSO-d6) δ9.13 (d, J=2.3Hz, 1H), 8.60 (dd, J=8.7, 2.3Hz, 1H), 8.18 (d, J=8.6Hz, 1H), 7. 52 (t, J=7.6Hz, 3H), 7.47 (d, J=8.4Hz, 3H), 7.42-7.37 (m, 3H), 7.37-7.32 (m, 3H), 7.22 (td, J=7.9, 7. 3, 1.6Hz, 2H), 7.17 (dd, J=7.6, 1.5Hz, 2H), 7.08 (dd, J=21.5, 8.4Hz, 4H), 7.00-6.92 (m, 4H), 3.91 (t, J=7.0Hz, 4H), 1.69 (h, J=6.6, 6.0Hz, 4H), 1.42 (h, J=7.4Hz, 4H), 1.34 (s, 6H), 0.90 (t, J=7.4Hz, 6H); 13 C NMR (150MHz, CDCl3) δ: 151.00, 149.20, 140.47, 139.25, 133.92, 132.91, 129.74, 129.4 1, 127.96, 127.79, 126.75, 126.12, 122.87, 120.38, 29.71, 20.16, 14.80, 13.84, 0.01.

[0024] Example 2

[0025] B-PHSD was made into 2×10 -5M PBS buffer solution (containing 20% THF). Dilute 10M sodium hydrosulfide solution with distilled water to concentrations of 0, 2, 4, 6, 8, 11, 14, 18, 22, 26, 30, 34, 38, 45 μM. Dilute 10M sodium hypochlorite solution with distilled water to concentrations of 0, 2, 4, 7, 10, 15, 25, 35, 45, 55, 70, 85, 100 μM. Sodium hydrosulfide solution of different concentrations was first added to the probe solution, and then sodium hypochlorite solution of different concentrations was added to the solution after the reaction was completed. The fluorescence emission spectrum of B-PHS-OP in the presence of different concentrations of hypochlorous acid was measured on a fluorescence spectrophotometer using fluorescence spectroscopy titration method, with an excitation wavelength of 580 nm, and the fluorescence emission peaks at 650 nm-850 nm were collected. From Figure 1 It can be seen that as the concentration of hydrogen sulfide in the solution gradually increases from 0M to 45μM, the emission peak of B-PHS-OP at 735nm gradually increases. Then, different concentrations of hypochlorous acid (0-100μM) were added to the solution. Figure 2 The emission peak of the B-PHS-OP solution at 685 nm begins to increase significantly. Therefore, this probe can be used as a near-infrared BODIPY-based fluorescent probe for sensitive and continuous detection of hydrogen sulfide and hypochlorous acid.

[0026] Example 3

[0027] B-PHS-OP was prepared into 2×10 -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 M solution. The fluorescence emission spectra of the probe at 735nm and 685nm were measured on a fluorescence spectrophotometer in the presence of different active oxygen, active sulfur, cations and anions using fluorescence spectrophotometry. The results are as follows Figure 3 As shown in Figure 2, only the addition of hydrogen sulfide can cause the fluorescence intensity of the probe to increase rapidly at 735 nm, while other analytes (Blank, Na + , K + , Cu 2+ , SO4 2- , SO3 2- , NO3 - , AcO - ,HCO3 - , N2H4, Cys, Hcy, GSH, TBHP, H2O2, NO, ONOO - , O2 - , 1 O2) and other references, the fluorescence intensity of the probe did not change significantly. Then, the analyte was added to the B-PHSD solution, such as Figure 4As shown in Figure 2, only the addition of hypochlorous acid can cause the fluorescence intensity of the probe to increase rapidly at 685 nm, while the other analytes (Blank, Na + , K + , Cu 2+ , SO4 2- , SO3 2- , NO3 - , AcO - ,HCO3 - , N2H4, Cys, Hcy, GSH, TBHP, H2O2, NO, ONOO - , O2 - , 1 Compared with the reference of HCl (H2O2), the fluorescence intensity of the probe did not change significantly. This shows that the compound can be used as a fluorescent probe for the specific and rapid continuous detection of hydrogen sulfide and hypochlorous acid.

Claims

1. A novel BODIPY-based near-infrared fluorescent probe for continuous detection of hydrogen sulfide and hypochlorous acid, characterized in that: The molecular formula of the probe is C 59 H 51 BF2N6O7S3, the structural formula is:

2. The method for preparing a novel BODIPY-based near-infrared fluorescent probe for continuous detection of hydrogen sulfide and hypochlorous acid according to claim 1, characterized in that: The steps include: 4-(3,7-bis(2-(10-butyl-10H-phenothiazin-3-yl)vinyl)-5,5-difluoro-1,9-dimethyl-5H-4,5-dipyrrolo[1,3,2]diazaborin-10-yl)phenol (abbreviated as B-PHS-OH) undergoes a substitution reaction with 2,4-dinitrobenzenesulfonyl chloride to obtain the probe 4-(3,7-bis(2-(10-butyl-10H-phenothiazin-3-yl)vinyl)-5,5-difluoro-1,9-dimethyl-5H-4,5-dipyrrolo[1,3,2]diazaborin-10-yl)phenyl 3,5-dinitrobenzenesulfonate, abbreviated as B-PHS-OP.

3. A novel BODIPY-based near-infrared fluorescent probe for continuous detection of hydrogen sulfide and hypochlorous acid according to claim 2, characterized in that: Under the action of triethylamine, B-PHS-OH undergoes a substitution reaction with 2,4-dinitrobenzenesulfonyl chloride to obtain the probe B-PHS-OP. The specific preparation method includes: (1) 15 mmol of B-PHS-OH and 5-10 mL of triethylamine were dissolved in anhydrous dichloromethane. 20-60 mmol of 2,4-dinitrobenzenesulfonyl chloride was then added to the mixture. The mixture was stirred at room temperature for 6 h. The reaction was monitored by TLC until the starting material spot disappeared, and the reaction was stopped. (2) 300-400 mL of dichloromethane was added to the reaction mixture, and then washed with distilled water. The organic phase was dried and distilled, and the resulting crude product was purified by silica gel column (dichloromethane: methanol = 150:1, v / v) to obtain compound B-PHS-OP as a brown solid.

4. Use of the novel BODIPY-based near-infrared fluorescent probe B-PHS-OP for continuous detection of hydrogen sulfide and hypochlorous acid according to claim 1 in the continuous detection of hydrogen sulfide and hypochlorous acid.

5. The use according to claim 4, characterized in that Probe B-PHS-OP reacts specifically and continuously with hydrogen sulfide and hypochlorous acid. When hydrogen sulfide is added to a solution of probe B-PHS-OP, the color of the solution rapidly changes from light blue to green under sunlight. Under 365nm ultraviolet light, the solution's fluorescence changes from colorless to dark red. Subsequently, when hypochlorous acid is added, the solution rapidly changes from green to cyan under sunlight. Under 365nm ultraviolet light, the fluorescence also rapidly changes from dark red to intense red.

6. The use according to claim 4, characterized in that B-PHS-OP can specifically and continuously identify the contents of hydrogen sulfide and hypochlorous acid, with detection limits as low as 89nM and 12nM, respectively.

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