阅读说明：本技术 一种用于检测HOCl的双分子荧光化合物及其制备与应用 (A kind of bimolecular fluorescent chemicals and its preparation and application for detecting HOCl ) 是由 朱勍 瞿姣姣 窦言东 金卉敏 于 2019-05-31 设计创作，主要内容包括：本发明涉及一种用于检测HOCl的双分子荧光化合物(I)及其制备与应用。本发明提供了一种新的双光子性质荧光探针化合物,可用于检测HClO,在生理条件下,该探针对HClO的选择性和敏感性优于其他相关分析物,包括活性氧、离子和生物硫醇。此外,该探针还成功地应用于双光子显微镜成像活细胞中的HClO,表明该探针在分析活细胞中的生物学功能和病理作用方面具有很高的潜力。<Image he="257" wi="700" file="DDA0002080968990000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>(The bimolecular fluorescent chemicals (I) and its preparation and application that the present invention relates to a kind of for detecting HOCl.The present invention provides a kind of new two-phpton property fluorescent probe compounds, can be used for detecting HClO, and in physiological conditions, which is better than other analyte of interest, including active oxygen, ion and biological thiol to the selectivity and sensibility of HClO.In addition, the probe is also successfully applied to the HClO in Two Photon Fluorescence imaging living cells, show that the probe has very high potentiality in terms of the biological function and pathological effect in analysis living cells.)

1. it is a kind of for detecting the bimolecular fluorescent chemicals of HOCl, shown in structure such as formula (I):

2. the method for preparing bimolecular fluorescent chemicals described in claim 1, which comprises with 2- first shown in formula (II) Base quinoline compound and the thioether benzaldehyde of formula (III) they are substrate, in the presence of anhydrous oxalic acid iron and trifluoroacetic acid, 100~ Bimolecular fluorescent chemicals shown in formula (I) are made in 120 DEG C of 10~18h of reaction；

3. method according to claim 2, it is characterised in that the reaction carries out in toluene solution.

4. method according to claim 2, it is characterised in that the 2- methylquinoline compound, thioether benzaldehyde, anhydrous two The ratio between amount of ferric acetate and trifluoroacetic acid substance is 1:1:0.05:0.05.

5. bimolecular fluorescent chemicals described in claim 1 are preparing the application in bimolecular fluorescence probe.

6. application as claimed in claim 5, it is characterised in that the bimolecular fluorescence probe is used to detect the hypochlorous acid of cell Value.

7. application as claimed in claim 6, it is characterised in that the cell is human cervical carcinoma cell Hela cell.

(1) technical field

The bimolecular fluorescent chemicals and its preparation and application that the present invention relates to a kind of for detecting HOCl.

(2) background technique

Hypochlorous acid (HOCl) is the important component for the anti-infection activity oxygen (ROS) that immunocyte generates, another party Face, the HOCl in homeostasis causes oxidative damage to biomolecule, more with inflammation, neurodegenerative disease, cardiovascular disease etc. Kind disease is related.In microorganism invasion, bactericidal effect has very great help to the autoimmune tool of main body.But due to The high reaction activity of HOCl and natural diffusivity, being difficult to control for HOCl product will will lead to a variety of diseases in phagocyte Generation, therefore, urgent need set up the method that HOCl can be accurately detected in living cells.

Popular direction of the fluorescent technique probe as chemical research in recent years, in view of the simple of fluorescent probe technique, efficiently, It is convenient, and cause the concern of more and more drug research persons, therefore it is quantitative to design a kind of highly sensitive colorimetric fluorescence probe Monitoring HClO value fluctuation is reasonably necessary and urgent, although there is some inspections for being used for HClO by fluorescent probe technique at present Survey method, but its preparation is all complex, and higher cost, lacks a kind of simple, effective method of modifying.

(3) summary of the invention

The bimolecular fluorescent chemicals and its preparation and application that it is an object of the present invention to provide a kind of for detecting HOCl.

The technical solution adopted by the present invention is that:

It is a kind of for detecting the bimolecular fluorescent chemicals of HOCl, shown in structure such as formula (I):

Two photon imaging has apparent advantage, and since its excitation wavelength is longer, penetration depth can be improved, reduce sample Product light injury.Wherein, two-photon is lacked since its excitation wavelength is longer, is increased penetration depth, is reduced the light loss of sample Wound.The compounds of this invention (I) is applied successfully to the HClO variation of single photon and Two Photon Fluorescence monitoring HeLa cell, table Its bright potential application foreground in tumour cell detection.

The invention further relates to the methods for preparing the bimolecular fluorescent chemicals, which comprises shown in formula (II) 2- methylquinoline compound and the thioether benzaldehyde of formula (III) be substrate, exist in anhydrous oxalic acid iron and trifluoroacetic acid Under, bimolecular fluorescent chemicals shown in formula (I) are made in 100~120 DEG C of 10~18h of reaction；

Specifically, the reaction carries out in toluene solution.

Preferably, the 2- methylquinoline compound, thioether benzaldehyde, anhydrous oxalic acid iron and trifluoroacetic acid substance The ratio between amount is 1:1:0.05:0.05.

The invention further relates to the bimolecular fluorescent chemicals to prepare the application in bimolecular fluorescence probe.Thioether is The recognition group of HClO can be used to detect hypochlorous acid by thioether and hypochlorous selective reaction.

Specifically, the bimolecular fluorescence probe is used to detect the hypochlorous acid value of cell.

Preferably, the cell is human cervical carcinoma cell Hela cell.

Compound (I) of the present invention can be used as two-photon fluorescence probe, the fluorescence detection applied to HClO.Described The method of the fluorescence detection of HClO concentration are as follows: using compound (I) as fluorescence probe, (5 μM) of HAPH displays one absorb extensively Band concentrates on 340nm (λ abs=340nm) and fluorescent belt concentrates on 430nm.Fluorescent emission band reduces at 470nm, while Occur a new transmitting band at 430nm, provides a kind of colorimetric method for the detection of ClO-；Next, in order to assess HAPH Selectivity, have detected probe to the fluorescence response of various bio-related molecules, the fluorescence response of probe can be ignored. Secondly, being incubated for HeLa after 0.5mM ClO- handles HAPH with probe HAPH can be observed the apparent color change of cell, in addition, In physiological conditions, probe shows that stable fluorescence exports to the variation of hypochlorous acid value, these results indicate that HAPH is to secondary Chlorate ions have preferable selectivity.

The beneficial effects are mainly reflected as follows: the present invention provides a kind of new two-phpton property fluorescence probes Object is closed, can be used for detecting HClO, in physiological conditions, which is better than other related point to sensibility to the selectivity of HClO Analyse object, including active oxygen, ion and biological thiol.It lives carefully in addition, the probe is also successfully applied to Two Photon Fluorescence imaging HClO in born of the same parents shows that the probe has very high potentiality in terms of the biological function and pathological effect in analysis living cells.

(4) Detailed description of the invention

Fig. 1 is the nucleus magnetic hydrogen spectrum of formula (I) compound in the present invention.

Fig. 2 is the absorption spectrum of formula (I) compound in the present invention.

Fig. 3 is the emission spectrum of formula (I) compound in the present invention.

Fig. 4 is the reality for the selectivity that formula (I) compound is directed to different ions in PBS (pH 7.4,10mM) in the present invention Test result；Figure small molecular is respectively hypochlorous acid, formaldehyde, formic acid, hydrogen peroxide, tert-butyl hydroperoxide, NaHS, paddy Guang Sweet peptide, cysteine, homocysteine, glucose.

Fig. 5 is the time gradient curve that formula (I) compound detects hypochlorous acid in the present invention.

Fig. 6 is ratio fluorescent image (the λ ex=that formula (I) compound probe-type (1) is incubated for Hela cell in the present invention 700nm)。

(5) specific embodiment

The present invention is described further combined with specific embodiments below, but protection scope of the present invention and not only limits In this: