Fluorescent dye containing azetidine spiro structure, and preparation method and application thereof

Abstract

The invention belongs to the technical field of fluorescence analysis, relates to a fluorescent dye containing a quaternary spiro structure, and a preparation method and application thereof, and concretely relates to a fluorescent dye containing an azetidine spiro structure, and a preparation method and application thereof. The fluorescent dye containing the azetidine spiro structure has a skeleton structure as shown in (I) or (II), and the fluorescent dye containing the azetidine spiro structure has excellent optical properties, good light stability and good biocompatibility. The invention further provides a novel azetidine spiro-structure-containing fluorescent dye with additional chemical connection sites, a preparation method thereof, and application of the novel azetidine spiro-structure-containing fluorescent dye in biological fluorescence analysis. The fluorescent molecule shows good water solubility, light stability and high fluorescence quantum yield, and can be used for fluorescence labeling, biological imaging and development of new fluorescent probes.

Description

technical field [0001] The invention belongs to the technical field of fluorescence analysis, and relates to a fluorescent dye containing a four-membered spiro ring structure, a preparation method and an application. It specifically relates to a fluorescent dye containing an azetidine spiro ring structure, and a preparation method and application thereof. Background technique [0002] The prior art discloses that fluorescence analysis technology has the advantages of high sensitivity, strong visualization, convenient operation, real-time detection of cells and organisms, etc., and has been widely used in ions, free radicals, molecules, polypeptides, enzymes, and even temperature, Identification of pH, etc.; small molecule fluorescent probes have been widely used in medicine, biology, environmental science and other disciplines for analysis, detection and biomarkers. [0003] Fluorescent dyes are an important part of small molecule fluorescent probes, and their photophysical...

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Problems solved by technology

[0004] With the deepening of life sciences, the detection of various active substances in the living body has become a research hotspot. The research practice shows that small molecule fluorescent detection is very important in terms of the selectivity, sensitivity, detection speed of detection substances, and the application of cells and living bodies. Needles also put forward higher requirements. However, the existing fluorescent dyes still have the defects of insufficient brightness and photostability for single molecule and live cell imaging.

Studies have disclosed that twisted intramolecular charge transfer (TICT) is one of the main non-radiative inactivation pathways in fluorescent molecules. In the TICT state, the dialkylamino donor generates a 90° twist left and right, forming a non-radiative, highly reactive chemical species, therefore, in order to avoid the formation of TICT, researchers have employed various modification strategies, such as Dylight, CF and Alexa dyes, which are commercially available fluorophores by making the amino Substituents were rigidized and complex structural modifications were carried out (J.Histochem.Cytochem., 1999,47,1179); Song Xiangzhi et al. used bulky 7-azacyclo[2.2.1]heptane to replace rhodamine dyes Dimethylamino substituent (J.Am.Chem.Soc., 2008, 130, 17652), a commonly used dimethylamino substituent, this structural modification greatly improves the quantum yield and photostability of rhodamine dyes, but its water solubility becomes poor; Lavis et al. N, N-difluorochrome in several types of fluorescent dyes such as naphthalimide, acridine compound, phenoxazine, coumarin, rhodamine, oxerhodamine, carbo-rhodamine and silyl-rhodamine The alkylamino substituent was replaced by azetidine, showing that the azetidine substitution significantly increased the brightness of the dye (Nat. Methods, 2015, 12, 244), this simple structural modification retained the physicochemical properties of the parent fluorophore , such as cell membrane permeability etc., have provided a good proof for the structural modification of fluorescent dyes to improve optical properties, however, studies have shown that the photostability of the fluorescent dyes substituted with azetidine is still not high (J.Am .Chem.Soc.,2019,141,981), therefore, it is necessary to further improve its photostability

[0005] The azetidine spiro ring structure is a special small molecule four-membered heterocyclic spiro ring, because the spiro ring structure relieves the electron density of the four-membered monocyclic ring and enhances the stability, and the introduction of the spiro ring can also be The multi-dimensional regulation of the properties of molecules such as lipidity, hydrophilicity, and spatial conformation has gradually attracted attention in drug research, but there have been no reports on fluorescent dyes based on the azetidine spiro ring structure.

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