Lysosome targeted pH fluorescent probe for monitoring cell autophagy as well as preparation and application thereof
A fluorescent probe and lysosome technology, applied in the field of pH fluorescent probes, can solve the problems of unsuitability for long-term detection, large cytotoxic side effects, strong background fluorescence, etc., and achieves good cell membrane permeability and easy mass production. , the effect of highly sensitive detection
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0031] 3',6'-bis(diethylamino)-2-(2-((2-(2-methoxyethoxy)ethyl)amino)ethyl)spiro[isoindoline-1, Preparation and characterization of 9'-xanthene]-3-one (RML):
[0032]
[0033] In a round bottom flask, 2-(2-aminoethyl)-3',6'-bis(diethylamino)spiro[isoindole-1,9'-xanthogen]-3-one (170mg , 0.62mmol) and 2-(2-methoxyethoxy) ethyl 4-methylbenzenesulfonate (300mg, 0.93mmol) were dissolved in N,N-dimethylformamide, heated to reflux for 12h to react Complete; the system was cooled to room temperature, and the solvent was removed by rotary evaporation under reduced pressure to obtain a crude product; the crude product was purified by silica gel column (eluent chloroform:methanol=20:1 by volume) to obtain a light yellow solid as the target compound RML.
[0034] 1 H NMR (600MHz, DMSO-d6 , Figure 1a )δ(ppm):1.11-1.13(t,12H,-CH3-),2.44(s,2H,-CH2-),2.64(s,2H,-CH2-),3.32-3.30(m,13H,- CH 2 -),3.47(m,4H,-CH 2 -),3.52(m,2H,-CH 2 -),6.23-6.24(d,2H,Ar-H),6.34(s,2H,Ar-H),6.39-6.40(d,2H,...
Embodiment 2
[0038] The fluorescent probe RML in Example 1 was prepared as a stock solution with a concentration of 1 mM in dimethyl sulfoxide and preserved. In the experiment, the probe was diluted to a final concentration of 25 μM with Britton-Robinson buffer (BR) at different pH values, and the UV absorption spectrum ( image 3 ). As the pH value decreased from 7.4 to 4.5, the absorption peak at 561nm gradually increased. At the same time, the color of the solution changed from colorless to pink ( Figure 4 ).
Embodiment 3
[0040] Also dilute the probe to a final concentration of 10 μM with Britton-Robinson buffer with different pH values, fix the excitation wavelength at 560 nm, and record the change of the fluorescent probe RML with pH in the DMSO / BR (1 / 99, v / v) system Fluorescence emission spectrum changes. As the pH value decreased from 7.4 to 4.4, the fluorescence intensity at 583nm gradually increased ( Figure 5 ). At the same time, the fluorescent color of the solution changed from colorless to red ( Figure 6 ). According to the Singmoidal fitting curve calculation of the fluorescence intensity value of the fluorescent probe RML at 583nm as a function of pH, the pKa value is 4.96 ( Figure 7 ), the pH response linear range is 4.5-5.7, which is very suitable for the detection of pH changes in the weakly acidic environment of lysosomes.
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com