Fluorescent probe for detecting CEES as well as synthesis method and application of fluorescent probe

Abstract

The invention discloses a fluorescent probe for detecting CEES as well as a synthesis method and application of the fluorescent probe, and belongs to the technical field of fluorescent probe detection. According to the novel fluorescent probe disclosed by the invention, a preparation method of # imgabs0 # is as follows: a # imgabs1 # probe reacts with CEES through a nucleophilic reaction to generate a sensing product, so that opening fluorescence response is generated, and the novel fluorescent probe has relatively high detection sensitivity. The problems that an existing fluorescent probe for detecting SM is unstable, false positive, short in fluorescence lifetime, short in emission wavelength and the like are solved.

Description

Technical Field

[0001] The invention belongs to the technical field of organic synthesis and fluorescent probe detection, and particularly relates to a fluorescent probe for detecting a mustard gas simulant CEES, and a synthesis method and application thereof. Background Art

[0002] Mustard gas or sulfur mustard (SM) is a well-known chemical warfare agent (CWAs). SM is chemically active and can cause poisoning damage to various tissues and organs such as the skin, eyes and respiratory tract of humans and animals. SM is also an alkylating agent that can undergo alkylation reactions with glutathione, proteins, DNA and many other biomacromolecules. Alkylation leads to DNA damage and glutathione depletion leads to oxidative stress, which are all related to the main toxicological mechanism of SM. In addition, SM has a simple structure and is easy to synthesize, making it a chemical terrorist threat. Moreover, it is still very harmful when left behind, and reports of injuries caused by accidental exposure to SM are common. After sulfur mustard is spread into the environment, it will remain active for a few hours to a few weeks depending on the ambient temperature and soil pH. At present, there is no specific antidote for clinical treatment, and the treatment of sulfur mustard poisoning can only be taken after the symptoms appear. It is worth noting that after exposure to SM, patients usually have an incubation period of 30 minutes to 8 hours before symptoms appear, which brings great difficulties to treatment. Therefore, it is very important to develop a sensitive and rapid method for mustard gas detection.

[0003] Fluorescent probes have been widely studied by scholars for their advantages such as low development cost, simple operation, high selectivity, and real-time imaging. Since SM and its analogs have low electrophilicity, the corresponding chemical sensors need to be designed as strong nucleophiles. Most of the existing fluorescent probes for detecting SM have problems such as instability, false positives, short fluorescence lifetime, and short emission wavelength. Therefore, it is necessary to further design and develop SM fluorescent probes with longer wavelengths and more stable properties. Summary of the invention

[0004] In order to overcome the shortcomings of the prior art, a fluorescent probe for detecting CEES with high specificity is provided. The present invention designs a novel fluorescent probe, which generates a sensing product by reacting with CEES through a nucleophilic reaction, thereby generating an on-fluorescence response, and has a high detection sensitivity. The fluorescent probe and synthesis method for detecting mustard gas simulating CEES described in the present invention use compound A as a raw material, and react with Fisher's aldehyde to obtain an intermediate product; then the intermediate product reacts with Lawesson's reagent to obtain the final product fluorescent probe.

[0005] A fluorescent probe for detecting mustard gas simulating CEES, the structural formula of the fluorescent probe is:

[0006]

[0007] Wherein, n in the structural formula is an integer of 0 to 2; X in the structural formula is O, S, NH, an alkoxy group containing 1 to 5 carbon atoms, an alkylthio group containing 1 to 5 carbon atoms, or an alkylamino group containing 1 to 5 carbon atoms; R1 and R2 in the structural formula are H at different positions on the benzene ring, hydrocarbons, olefins, alkynyl groups, aromatic groups, hydroxyl groups, halogens, nitro groups, benzyloxy groups, alkoxy groups, benzene rings, or various water-soluble groups (e.g., SO3M); R3 in the structural formula is methyl, an alkyl group containing 3 to 20 carbon atoms, sulfonic acid, phosphoric acid, carboxylic acid, and carboxylic acid derivatives.

[0008] The preparation method comprises the following steps:

[0009] Step 1, compound A and Fischer's aldehyde B (0.2 to 2 times the amount of feed) are dissolved in a solvent, 0.01 to 5 times of a base is used as a catalyst, and 0.5 to 3 times of N, N-dicyclohexylcarboximide is used as a dehydrating agent, the above multiples are based on compound A, react at 50 to 140° C. for 10 to 40 hours to obtain a crude product D, remove the solvent, and obtain a pure compound D through column chromatography;

[0010] Step 2, compound D and Lawesson's reagent E (0.2 to 4 times the amount of feed) are dissolved in a solvent, protected by argon gas, and the above multiples are based on compound D. The reaction is carried out at 50 to 140° C. for 5 to 60 minutes to obtain a crude product F. The solvent is removed and the pure compound F is obtained by column chromatography, which is a fluorescent probe for detecting mustard gas simulating CEES.

[0011]

[0012] Further in step 1; the solvent required for the reaction is benzene, toluene, etc., and the base required is piperidine, pyridine, triethylamine, etc.;

[0013] In the further step 2, the solvent required for the reaction is benzene, toluene, etc.

[0014] Synthesis route:

[0015]

[0016] The probe of the invention is used for detecting mustard gas and mustard gas simulants.

[0017] The fluorescent probe of the method of the invention is easy to prepare, has a short synthesis route, mild reaction conditions and is suitable for large-scale production.

[0018] The CEES fluorescent probe prepared by the invention has good stability and specificity to CEES. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1The UV (left) and fluorescence (right) spectra of the probe Cy3C=S after interacting with CEES;

[0020] Figure 2 The compound Cy3C=O 1 H NMR spectrum;

[0021] Figure 3 The compound Cy3C=O 13 C NMR spectrum;

[0022] Figure 4 is the HRMS spectrum of compound Cy3C=O;

[0023] Figure 5 The compound Cy3C=S 1 H NMR spectrum;

[0024] Figure 6 The compound Cy3C=S 13 C NMR spectrum;

[0025] Figure 7 It is the HRMS spectrum of compound Cy3C=S. DETAILED DESCRIPTION

[0026] The following specific embodiments will help to understand the present invention, but will not limit the contents of the present invention.

[0027] Example 1

[0028] This embodiment is a near-infrared fluorescent probe Cy3C=S for mustard gas simulated CEES detection, and the synthesis method is:

[0029]

[0030] Compound A (628 mg) and Fischer's aldehyde B (500 mg) were dissolved in 10 mL of toluene, piperidine (700 μL) and N,N-dicyclohexylcarboximide (265 mg) were added, and the mixture was heated under reflux for 30 hours. Pure compound Cy3C=O was obtained by silica gel column chromatography with a yield of 74%.

[0031] 1H NMR(400MHz,Chloroform-d)δ8.20(dd,J=7.9,1.6Hz,1H),7.90(dd,J=14.6,12.4Hz,1H),7.68–7.60(m,1H),7.48(d,J=8.3Hz,1H),7.40–7.33(m,1H), 7.27–7.19(m,2H),6.96(t,J=7.4Hz,1H),6.73(d,J=7.8Hz,1H),6.16(s,1H ),5.96(d,J=14.6Hz,1H),5.51(d,J=12.4Hz,1H),3.23(s,2H),1.69(s,4H). 13 C NMR(101MHz,Chloroform-d)δ178.00,164.19,162.80,155.98,144.49,138.98,132.91,127.97,125.5 3,124.49,124.29,121.69,120.79,117.49,113.27,106.79,106.49,95.44,46.36,29.30,28.72.HRMS m / z:Calcd for C 23 H 21 NO2[M+H] + ,344.1645;found,344.1640.

[0032] Compound Cy3C=O (500 mg) and Lawesson's reagent E (323 mg) were dissolved in 15 mL of super dry benzene, protected by argon, heated under reflux for 30 minutes, and purified by silica gel column chromatography to obtain pure compound Cy3C=S with a yield of 47%.

[0033] 1 H NMR(400MHz,Chloroform-d)δ8.55(d,J=8.0Hz,1H),8.08–7.99(m,1H),7.65–7.58(m,1H),7.42(d,J=8.0Hz,1H),7.36(d,J=8.0Hz,1H),7 .27–7.22(m,3H),7.00(t,J=7.2Hz,1H),6.79(d,J=7.8Hz,1H),5.92(d,J=14.3Hz,1H),5.64(d,J=12.7Hz,1H),3.29(s,3H),1.67(s,6H). 13C NMR(101MHz,Chloroform-d)δ164.81,156.78,151.40,144.17,139.27,137.89,133.00,128.91,128.7 2,128.12,125.36,121.81,121.69,120.93,117.51,111.97,107.50,97.14,47.00,29.70,28.67.HRMS m / z:Calcd for C 23 H 21 NOS[M+H] + ,360.1416;found,360.1413.

[0034] Example 2

[0035] The mustard gas-simulating CEES fluorescent probe Cy3C=S prepared in Example 1 was dissolved in methanol / isopropyl ether (1:9), and its absorption spectrum was measured using a UV-visible spectrophotometer. Figure 1 (Left) As shown, the maximum ultraviolet absorption of the probe Cy3C=S itself is 535nm. When CEES is added to the probe solution, the absorption peak at 535nm decreases and a new absorption peak is generated at 653nm.

[0036] The fluorescence spectrum of the compound in methanol / isopropyl ether (1:9) was tested using a fluorescence spectrometer. Figure 1 As shown in (right), the probe Cy3C=S produces almost no fluorescence in the range of 625-850nm, but when the probe interacts with CEES, it produces strong fluorescence at 655nm, and the color changes from purple to blue.

Claims

1. A fluorescent probe for detecting mustard gas simulating CEES, characterized in that The structural formula of the fluorescent probe is: Wherein, n in the structural formula is an integer of 0 to 2; X in the structural formula is O, S, NH, an alkoxy group containing 1 to 5 carbon atoms, an alkylthio group containing 1 to 5 carbon atoms, or an alkylamino group containing 1 to 5 carbon atoms; R1 and R2 in the structural formula are H at different positions on the benzene ring, hydrocarbons, olefins, alkynyl groups, aromatic groups, hydroxyl groups, halogens, nitro groups, benzyloxy groups, alkoxy groups, benzene rings, or various water-soluble groups (e.g., SO3M); R3 in the structural formula is methyl, an alkyl group containing 3 to 20 carbon atoms, sulfonic acid, phosphoric acid, carboxylic acid, and carboxylic acid derivatives.

2. A method for synthesizing a fluorescent probe for detecting mustard gas simulating CEES according to claim 1, characterized in that: The details include: Step 1, compound A and Fischer's aldehyde B (0.2 to 2 times the amount of feed) are dissolved in a solvent, 0.01 to 5 times of a base is used as a catalyst, and 0.5 to 3 times of N, N-dicyclohexylcarboximide is used as a dehydrating agent, the above multiples are based on compound A, react at 50 to 140° C. for 10 to 40 hours to obtain a crude product D, remove the solvent, and obtain a pure compound D through column chromatography; Step 2, compound D and Lawesson's reagent E (0.2 to 4 times the amount of feed) are dissolved in a solvent, protected by argon, and the above multiples are based on compound D, reacting at 50 to 140° C. for 5 to 60 minutes to obtain a crude product F, removing the solvent, and subjecting the mixture to column chromatography to obtain a pure compound F, i.e., a fluorescent probe for detecting mustard gas simulating CEES; Synthesis route:

3. The method according to claim 2, wherein the solvent required for the reaction in step 1 is benzene, toluene, etc., and the base required is piperidine, pyridine, triethylamine, etc.; In step 2, the solvent required for the reaction is benzene, toluene, etc.

4. The fluorescent probe for detecting mustard gas simulating CEES according to claim 1, characterized in that: Its absorption spectrum wavelength ranges from 350 to 850 nm.

5. The use of the fluorescent probe for detecting mustard gas simulating CEES according to claim 1 is used for the detection of mustard gas and mustard gas simulants.

Images