A fluorescent probe for detecting cees and its synthesis method and application

Abstract

The application relates to a fluorescent probe for detecting CEES and a synthesis method and application thereof, and belongs to the technical field of fluorescent probe detection. A novel fluorescent probe is prepared by the method: the probe is reacted with CEES through a nucleophilic reaction to generate a sensing product, thereby generating an open fluorescent response, and the fluorescent probe has high detection sensitivity. The application solves the problems of instability, false positive, short fluorescent lifetime and short emission wavelength of the existing fluorescent probe for detecting SM.

Description

TECHNICAL FIELD

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

[0002] Mustard gas or sulfur mustard (SM) is a famous chemical warfare agent (CWA). SM has chemical activity and can cause poisoning damage to various tissues and organs such as skin, eyes and respiratory tract of humans and animals. SM is also an alkylating agent that can undergo alkylation with glutathione, proteins, DNA and many other biological macromolecules. Alkylation leads to DNA damage and glutathione depletion, which are related to the main toxicological mechanism of SM. In addition, SM is simple in structure and easy to synthesize, and becomes a chemical terrorist threat. Moreover, it still has great harm left over, and reports of injuries caused by accidental contact with SM are not uncommon. After sulfur mustard is dispersed into the environment, it can remain active for a few hours to a few weeks depending on the environmental temperature and soil pH. At present, there is no specific antidote for clinical treatment, and the treatment of sulfur mustard poisoning can only take corresponding measures after the symptoms appear. It is worth noting that after contacting SM, the patient usually has a latent period of 30 minutes to 8 hours before symptoms appear, which brings great difficulty to treatment. Therefore, it is very important to develop a sensitive and rapid method for detecting mustard gas.

[0003] Fluorescent probes are widely studied by scholars due to their 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 wavelength and more stable properties. SUMMARY

[0004] In order to overcome the deficiencies of the prior art, a fluorescent probe with high specificity for detecting CEES is provided, and a new type of fluorescent probe is designed. The new type of fluorescent probe generates a sensing product through a nucleophilic reaction with CEES, thereby producing an on-off fluorescent response, and has high detection sensitivity. The fluorescent probe for detecting mustard gas analog CEES and the synthesis method disclosed by the application use compound A as a raw material, and add Fischer's aldehyde to obtain an intermediate product through an addition-elimination reaction; then the intermediate product is reacted with Lawson's reagent to obtain the final product fluorescent probe.

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

[0006]

[0007] wherein n in the structural formula is an integer from 0 to 2; X in the structural formula is O, S, NH, alkoxy containing 1-5 carbons, alkylthio containing 1-5 carbons, alkylamino containing 1-5 carbons; R1 and R2 in the structural formula are H, hydrocarbon, alkene, alkyne, aryl, hydroxyl, halogen, nitro, benzyloxy, alkoxy, benzene ring or various water-soluble groups (for example: SO3M) at different sites on the benzene ring; and R3 in the structural formula is methyl, alkyl containing 3-20 carbons, sulfonic acid, phosphoric acid, carboxylic acid and carboxylic acid derivatives.

[0008] The preparation method comprises the following steps: specifically comprising the following:

[0009] Step 1, compound A is dissolved in a solvent with Fisher aldehyde B (0.2-2 times the amount of raw material), 0.01 times-5 times of a base is used as a catalyst, and 0.5 times-3 times of N, N-dicyclohexyl carbodiimide is used as a dehydrating agent, the above times are based on compound A, and the crude product D is obtained by reacting at 50-140 DEG C for 10-40 hours, the solvent is removed, and column chromatography is performed to obtain pure compound D;

[0010] Step 2, compound D is dissolved in a solvent with Lawesson reagent E (0.2-4 times the amount of raw material), and argon protection is performed, the above times are based on compound D, and the crude product F is obtained by reacting at 50-140 DEG C for 5-60 minutes, the solvent is removed, and column chromatography is performed to obtain pure compound F, that is, a fluorescent probe for detecting mustard gas simulation CEES.

[0011]

[0012] In further step 1, the required solvent for the reaction is benzene, toluene and the like, and the required base is piperidine, pyridine, triethylamine and the like;

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

[0014] Synthetic route:

[0015]

[0016] The probe described in the application is used for detecting mustard gas and mustard gas simulation agents.

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

[0018] The CEES fluorescent probe prepared in the application has good stability and specificity for CEES. BRIEF DESCRIPTION OF DRAWINGS

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

[0020] Figure 2 is the H NMR spectrum of the compound Cy3C=O; 1

[0021] Figure 3 is the C NMR spectrum of the compound Cy3C=O; 13

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

[0023] Figure 5 is the H NMR spectrum of the compound Cy3C=S; 1

[0024] Figure 6 is the C NMR spectrum of the compound Cy3C=S; 13

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

[0026] The present application will be understood more clearly by the following detailed description, but is not limited by the content of the detailed description.

[0027] Example 1

[0028] The present embodiment is a near-infrared fluorescent probe Cy3C=S for detecting mustard gas analog CEES, and the synthesis method is as follows:

[0029]

[0030] Compound A (628 mg) and Fischer aldehyde B (500 mg) were dissolved in 10 mL of toluene, and piperidine (700 μL) and N,N-dicyclohexylcarbodiimide (265 mg) were added, and heated to reflux for 30 hours, and then column chromatography on silica gel was performed to obtain pure compound Cy3C=O, with a yield of 74%.

[0031] 1 ​​​​H NMR (400 MHz, Chloroform-d) δ 8.20 (dd, J = 7.9, 1.6 Hz, 1H), 7.90 (dd, J = 14.6, 12.4 Hz, 1H), 7.68 - 7.60 (m, 1H), 7.48 (d, J = 8.3 Hz, 1H), 7.40 - 7.33 (m, 1H), 7.27 - 7.19 (m, 2H), 6.96 (t, J = 7.4 Hz, 1H), 6.73 (d, J = 7.8 Hz, 1H), 6.16 (s, 1H), 5.96 (d, J = 14.6 Hz, 1H), 5.51 (d, J = 12.4 Hz, 1H), 3.23 (s, 2H), 1.69 (s, 4H). 13 C NMR (101 MHz, Chloroform-d) δ 178.00, 164.19, 162.80, 155.98, 144.49, 138.98, 132.91, 127.97, 125.53, 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=0 (500 mg) and Lawesson's reagent E (323 mg) were dissolved in 15 mL of super dry benzene, under argon protection, heated to reflux for 30 minutes, and the pure compound Cy3C=S was obtained by silica gel column chromatography with a yield of 47%.

[0033] 1 H NMR (400 MHz, Chloroform-d) δ 8.55 (d, J = 8.0 Hz, 1H), 8.08 - 7.99 (m, 1H), 7.65 - 7.58 (m, 1H), 7.42 (d, J = 8.0 Hz, 1H), 7.36 (d, J = 8.0 Hz, 1H), 7.27 - 7.22 (m, 3H), 7.00 (t, J = 7.2 Hz, 1H), 6.79 (d, J = 7.8 Hz, 1H), 5.92 (d, J = 14.3 Hz, 1H), 5.64 (d, J = 12.7 Hz, 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-simulated CEES fluorescent probe Cy3C=S prepared in Specific Example 1 was dissolved in methanol / isopropyl ether (1:9), and its absorption spectrum was measured using a UV-Vis spectrophotometer, as detailed in the attached figure. Figure 1 As shown on the left, the maximum UV absorption of the probe Cy3C=S 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 this compound in methanol / isopropyl ether (1:9) was measured using a fluorescence spectrometer, as detailed in the attached figure. Figure 1 As shown on the right, the probe Cy3C=S produces almost no fluorescence in the 625-850nm range. 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-simulated CEES, characterized in that, The structural formula of the fluorescent probe is: 。 2. The method for synthesizing a fluorescent probe for detecting mustard gas-simulated CEES according to claim 1, characterized in that, Specifically, it includes the following: Step 1: Compound A is dissolved in a solvent with 0.2 to 2 times the amount of Fisher's aldehyde B. 0.01 to 5 times the amount of base is used as a catalyst, and 0.5 to 3 times the amount of N,N-dicyclohexylcarboimide is used as a dehydrating agent. The above multiples are based on compound A. The reaction is carried out at 50 to 140°C for 10 to 40 hours to obtain crude product D. The solvent is removed, and pure compound D is obtained by column chromatography. Step 2: Compound D is dissolved in a solvent with 0.2 to 4 times the amount of Lawson's reagent E, under argon protection. 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 crude product F. The solvent is removed, and pure compound F is obtained by column chromatography, which is the fluorescent probe for detecting mustard gas simulating CEES. Synthesis route: 。 3. According to claim 2, the solvent required for the reaction in step 1 is benzene or toluene, and the base required is piperidine, pyridine, or triethylamine; The solvents required for the reaction in step 2 are benzene and toluene.

4. The application of the fluorescent probe for detecting mustard gas mimicking CEES according to claim 1, for the detection of mustard gas and mustard gas mimics for non-diagnostic and non-therapeutic purposes.

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