CN-122011027-A - Preparation and application of hydrogen peroxide fluorescent probe based on furonitrile-xanthene

Abstract

The invention designs synthesis and application of a furonitrile-xanthene-based hydrogen peroxide fluorescent probe, which has the structural formula: The invention provides a preparation method for synthesizing a fluorescent probe by taking a compound FX and diphenyl phosphoryl chloride as raw materials, wherein the fluorescent probe is a hydrogen peroxide (H 2 O 2 ) fluorescent probe with high sensitivity, high selectivity and low cytotoxicity, firstly, the fluorescent probe has better sensitivity to the fluorescent response of the system H 2 O 2 change, the linear range is 1 mu M-70 mu M, secondly, the fluorescent probe is not influenced by other various cations, anions, amino acids, biological thiols and active oxygen, only shows good selectivity to H 2 O 2 , and in addition, the fluorescent probe has rapid action with H 2 O 2 , the response time is within 10min, when the pH value is 7.0-8.0, the fluorescent response signal of the probe is strongest, which indicates that the fluorescent probe can detect H 2 O 2 under a complex physiological environment, and in addition, the fluorescent probe has small cytotoxicity and can be applied to the detection of living cells H 2 O 2 .

Inventors

• LI CHUNYAN
• XU JIA
• CHEN JING

Assignees

• 湘潭大学

Dates

Publication Date

20260512

Application Date

20260318

Claims (3)

1. 1. A furonitrile-xanthene-based hydrogen peroxide fluorescent probe, FX-P, characterized by the following structure: 。
2. 2. The preparation of a furonitrile-xanthene-based hydrogen peroxide fluorescent probe according to claim 1, characterized by the following reaction steps: Sequentially adding 1.2-1.5 equivalent of compound FX and 1.0 equivalent of 4-dimethylaminopyridine into a 50 mL round bottom flask at a temperature of minus 4 ℃, then adding 20-30 mL of tetrahydrofuran, adding 100 mu L of triethylamine under the protection of nitrogen, stirring for 10-30 min, then adding 1.0 equivalent of diphenyl phosphoryl chloride, reacting for 1-3 h, gradually heating to room temperature, reacting for 8-12 h, removing solvent from the crude product under the condition of reduced pressure after the reaction is finished, and purifying by column chromatography with methylene dichloride/methanol eluent with the volume ratio of 100:1-50:1 to obtain dark purple solid FX-P, namely the fluorescent probe.
3. 3. The use of a furonitrile-xanthene based hydrogen peroxide fluorescent probe according to claim 1, wherein the fluorescent probe is applicable for the detection of hydrogen peroxide in living cells.

Description

Preparation and application of hydrogen peroxide fluorescent probe based on furonitrile-xanthene Technical Field The invention belongs to the technical field of fluorescent probes, and particularly relates to preparation and application of a furonitrile-xanthene-based hydrogen peroxide fluorescent probe. Background Most of the oxidation process （Wang S, Yao J, Wang B, et al. A ratiometric and two-photon fluorescent probe for imaging hydrogen peroxide in living cells [J]. Luminescence 2022, 37, 1037–1043.）,, in which hydrogen peroxide (H 2O2) is widely present in biological systems, is closely related to many physiological and pathological processes. As an important active oxygen and intracellular second messenger, H 2O2 is not only an oxidative stress marker, but also plays a key role in redox signal paths and immune response, and many researches on regulating cell proliferation, differentiation, migration and disease development （Peng Z, Cui M, Chu J, et al. A novel AIE fluorescent probe for the detection and imaging of hydrogen peroxide in living tumor cells and in vivo [J]. Bioorganic Chemistry 2024, 150, 107592; Ren M, Deng B, Zhou K, et al. Single fluorescent probe for dual-imaging viscosity and H2O2 in mitochondria with different fluorescence signals in living cells [J]. Analytical Chemistry 2017, 89, 552–555.）. show that the level of H 2O2 in cells is closely related to various diseases such as cardiovascular diseases, neurodegenerative diseases and liver injury （Qiu X, Xin C, Qin W, et al. A novel pyrimidine based deep-red fluorogenic probe for detecting hydrogen peroxide in Parkinson's disease models [J]. Talanta 2019, 199, 628–633; Fan X, Sun Y, Fu J, et al. MRI-responsive nanoprobes for visualizing hydrogen peroxide in diabetic liver injury [J]. Biomaterials, 2025, 321, 123292.）. because hydrogen peroxide has important physiological and clinical significance, and it is important to design an effective method to realize accurate detection in a complex biological system. In recent years, various analytical methods for detecting hydrogen peroxide, including chromatography, electrochemical methods, spectrophotometry, and the like, have been reported successively by researchers. The sample pretreatment operations of the methods are complicated, and the dynamic change characteristics （Zhou Y, Yang X, Lee H, et al. Small-molecule fluorescent probes for detecting hydrogen peroxide in biological systems [J]. Coordination Chemistry Reviews, 2025, 541: 216785）. of hydrogen peroxide in the body cannot be effectively reflected, so that it is important to detect the hydrogen peroxide by adopting an effective method. Fluorescent imaging technology has received much attention （Zhou R, Sha H, Fu S, et al. Hyperspectral Fluorescence Imaging with a New Polarity-Ultrasensitive Fluorescent Probe[J]. Advanced Science, 2025, e08792.）. from researchers because of its high sensitivity, high resolution, simplicity and real-time detection capability, and has gradually become an important research tool （Wang F, Zhong Y, Bruns O, et al. In vivo NIR-II fluorescence imaging for biology and medicine[J]. Nature Photonics, 2024, 18, 535-547.）. in the fields of chemistry, biology and medicine, so far, many fluorescent probes have been developed to detect H2O2（Li W, Fu T, Zheng M, et al. Discovery of a highly selective fluorescent probe for hydrogen peroxide and its biocompatibility evaluation and bioimaging applications in cells and zebrafish [J]. Bioorganic Chemistry, 2024, 150, 107552; Peng Z, Cui M, Chu J, et al. A novel AIE fluorescent probe for the detection and imaging of hydrogen peroxide in living tumor cells and in vivo [J]. Bioorganic Chemistry, 2024, 150, 107592.）., but these probes have some problems of short fluorescence emission wavelength and weak tissue penetration. Therefore, it is very interesting to design and synthesize near infrared fluorescent probes with long wavelength emission that can achieve high sensitivity detection of hydrogen peroxide. Furonitrile-xanthene is taken as a novel fluorescent dye, has long emission wavelength, thus shows deeper tissue penetration depth and is more beneficial to biological imaging. There is currently no probe based on a furannitrile-xanthene dye to detect H 2O2. Thus, the present invention designs and synthesizes a furonitrile-xanthene based fluorescent probe to detect hydrogen peroxide. Disclosure of Invention In light of the requirements set forth, the inventors have conducted intensive studies on this, and after a great deal of creative effort, they have provided a furonitrile-xanthene-based hydrogen peroxide fluorescent probe. The technical scheme of the invention is that the hydrogen peroxide fluorescent probe based on the furonitrile-xanthene has the following structure: a preparation method of a furonitrile-xanthene-based hydrogen peroxide fluorescent probe comprises the following steps: Sequentially adding 1.2-1.5 equivalent of compound FX and 1.0 equivalent of 4-d