A kind of hypoxia fluorescent probe and its application
A fluorescent probe and probe technology, which is applied in the field of cyclic hypoxia fluorescent probes and hypoxia fluorescent probes, can solve the problems of inability to detect cyclic hypoxia in real time and dynamically, and achieve high selectivity, fast response speed, and wide application foreground effect
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Embodiment 1
[0043] Embodiment 1: the preparation of compound 2
[0044]
[0045] Compound 2 (F.Kong, Z.Liang, D.Luan, X.Liu, K.Xu, and B.Tang, A Glutathione (GSH)-Responsive Near-Infrared (NIR) Theranostic Prodrug for Cancer Therapy was synthesized by known methods in the literature and Imaging Anal. Chem. 2016, 88, 6450-6456.).
[0046] Synthesis of compound 1: m-nitrophenol (7.61mmol, 1.059g) and NaH (7.61mmol, 0.304g) were dissolved in anhydrous DMF solution (80mL), stirred at room temperature under Ar atmosphere for 10min. IR780 (3mmol, 2g) was dissolved in anhydrous DMF (10mL), slowly added dropwise into the above solution with a syringe, reacted at room temperature in the dark for 24h, followed by mass spectrometry. Solvent removal by rotary evaporation, purification by silica gel column, developing solvent: CH 2 Cl 2 :CH 3 OH=50:1 (including 2‰TEA), 1.4 g of green solid was obtained, and the yield was about 60%.
[0047] Synthesis of Compound 2: SnCl 2 2H 2 O (36.411mmol,...
Embodiment 2
[0048] Embodiment 2: the preparation of HDMA probe (be the probe of electron-donating group on the azophenyl group)
[0049]
[0050] HDMA probes were synthesized by methods known in the literature (Tian, X., Li, Z., Sun, Y., Wang, P., Ma, H. Near-Infrared Fluorescent Probes for Hypoxia Detection via Joint Regulated Enzymes: Design, Synthesis, and Application in Living Cells and Mice. Anal Chem. 2018, 90, 13759-13766.). Compound 2 (83.6μmol, 45mg) was dissolved in acetonitrile / dichloromethane (5mL, 1:4, v / v) mixed solvent containing 2% (0.1mL, v / v) trifluoroacetic acid (TFA), Ar atmosphere Stir at 0°C, add sodium nitrite (162.7 μmol, 15 mg), stir at 0°C for 15 minutes, TCL detects that the raw material point disappears, add sulfamic acid (155.24 μmol, 15 mg), and continue stirring at 0°C for 5 minutes. N,N-Dimethylaniline (538.9 μmol, 65.3mg) was dissolved in acetonitrile solution (1mL), slowly added to the above solution in batches, reacted at 0°C for 1h, and detected b...
Embodiment 3
[0051] Embodiment 3: the preparation of HDSF probe
[0052]
[0053] 3,5-bis(trifluoromethyl)aniline (11.15mmol, 2.54g) was dissolved in dichloromethane (75mL) solution and stirred rapidly. Oxone (22.3mmol, 13.712g) was dissolved in secondary water (112mL) solution, added dropwise to the above dichloromethane solution, and stirred at room temperature for 22h in the dark. Separate the liquids, extract the aqueous phase with dichloromethane (50mL x 2), combine the organic phases, wash with 0.1M dilute hydrochloric acid (50mL x 2) and secondary water (50mL x 2), dry over anhydrous sodium sulfate, and reduce pressure The solvent was removed by rotary evaporation to obtain a green liquid (3,5-bis(trifluoromethyl)nitrosobenzene). This product was unstable and was directly used in the next step.
[0054] Compound HD-NH 2 (0.7mmol, 380mg) was dissolved in glacial acetic acid (260mL), added the above green product, stirred at room temperature in the dark, and followed the reaction...
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