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Nitroreductase fluorescent probe based on nitro reduction and sulfur-nitrogen transposition, and preparation method thereof

A technology of nitroreductase and fluorescent probe, which is applied in the field of chemical analysis to achieve the effects of low biological toxicity, specific fluorescence imaging detection, sensitive and selective detection

Active Publication Date: 2020-04-28
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, a variety of nitroreductase fluorescent probes have been reported successively, but the nitroreductase probes of the prior art are still designed based on the domino decomposition reaction or the electronic push-pull effect after the nitro group is reduced to an amino group, and it is not yet possible To meet the needs of research and practical operation

Method used

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  • Nitroreductase fluorescent probe based on nitro reduction and sulfur-nitrogen transposition, and preparation method thereof
  • Nitroreductase fluorescent probe based on nitro reduction and sulfur-nitrogen transposition, and preparation method thereof
  • Nitroreductase fluorescent probe based on nitro reduction and sulfur-nitrogen transposition, and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Synthetic probe (1), its structure is:

[0045]

[0046] The synthesis technique of probe (1) is:

[0047]

[0048] Compound C (122mg, 0.338mmol) and Compound e (105mg, 0.667mmol) were dissolved in dry dichloromethane, DMAP (124mg, 1.016mmol) was added, stirred at room temperature for 5 hours under nitrogen protection, the reaction solution was concentrated and passed through a silica gel column Chromatography (petroleum ether: ethyl acetate: dichloromethane = 20:1:1) gave compound (1), 62 mg, yield 38.5%. LC-MS (ESI) m / z: 477.1 [M+H] + ; 1 H NMR (400MHz, DMSO-d 6 )δ8.21-8.15(m, 1H), 7.60-7.48(m, 4H), 7.46-7.42(m, 2H), 7.35(t, J=7.8Hz, 1H), 6.90(d, J=8.2Hz , 1H), 6.57(d, J=3.9Hz, 1H), 6.26(d, J=3.8Hz, 1H), 2.47(s, 3H), 2.30(q, J=7.6Hz, 2H), 1.38(s , 3H), 0.90(t, J=7.5Hz, 3H); 13 CNMR(151MHz,Chloroform-d)δ165.83,144.24,142.40,139.71,138.80,137.04,136.59,135.92,133.02,132.67,128.89,128.78,128.18,127.97,124.85,124.57,124.23,123.33,16.51,13.44, 13.02, 11.79.

Embodiment 2

[0049] Example 2. Changes in UV absorption spectra before and after the reaction of probe (1) with nitroreductase

[0050] NADH is formulated into 0.15M Tris solution, which is ready-to-use; the probe is formulated into 3mM acetonitrile solution, which is used for subsequent use; nitroreductase (1mg, 90%) is prepared into a 3g / L stock solution with deionized water , aliquoted and stored at -20°C for later use. Add 3 mL of Tris (pH 7.4, 50 mM) buffer solution containing 25% acetonitrile into a 3 mL quartz cuvette, then add 10 μL of probe, 10 μL of NADH and 5 μL of nitroreductase in sequence. React at 37°C for 180min, and measure its absorption spectrum. Depend on image 3 It can be seen that the maximum ultraviolet absorption intensity at 505 nm of the probe (1) becomes weaker before and after the reaction.

Embodiment 3

[0051] Embodiment 3. Fluorescent emission spectrum changes before and after the probe (1) reacts with nitroreductase

[0052] Add 3 mL of Tris (50 mM, pH 7.4) buffer solution containing 25% acetonitrile to a 3 mL quartz cuvette, then add 10 μL of probe, 10 μL of NADH and 5 μL of nitroreductase (NTR) in sequence. React at 37°C for 180 min, and measure its fluorescence emission spectrum. The excitation wavelength is 470nm, the excitation slit width is 10nm, the emission slit width is 10nm, and the gain is 700V. Depend on Figure 4 It can be seen that the fluorescence of the probe (1) itself is weak, and the maximum fluorescence emission is 560 nm after reacting with nitroreductase, and the fluorescence is enhanced by about 7 times.

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Abstract

The invention belongs to the technical field of chemical analysis, and relates to a small molecular fluorescent probe, particularly to a nitroreductase fluorescent probe based on nitro reduction and sulfur-nitrogen transposition, wherein the fluorescent probe is a BODIPY structure nitroreductase fluorescent probe. The invention discloses preparation and applications of the probe and a novel reaction mechanism for nitroreductase recognition. The fluorescent probe disclosed by the invention is stable in photophysical activity and high in nitroreductase response sensitivity, and can be applied tofluorescence imaging detection of hypoxic tumor cells.

Description

technical field [0001] The invention belongs to the technical field of chemical analysis, and relates to a small molecule fluorescent probe, in particular to a nitroreductase fluorescent probe based on nitro reduction and sulfur nitrogen translocation, and its preparation method and application, especially a fluoroborate dipyrrole (BODIPY) structural class nitroreductase fluorescent probe and its preparation method and application. Background technique [0002] The prior art discloses that hypoxia is related to various diseases such as cardiac ischemia, stroke, inflammatory diseases, tumors, etc., especially in solid tumors where the oxygen content is less than 4.4%, and the hypoxic state of the tumor is directly related to the development and metastasis of the tumor , drug resistance and treatment tolerance. [0003] Studies have shown that nitroreductase will be overexpressed in tissues or cells under hypoxic conditions. Nitroreductase can be used as an important indicato...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09K11/06C07F5/02G01N21/64G01N21/33
CPCC09K11/06C07F5/022G01N21/643G01N21/6428G01N21/33C09K2211/1007C09K2211/1029C09K2211/1055
Inventor 古险峰李咪咪高洁
Owner FUDAN UNIV
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