Design and Synthesis of Chemiluminescent Probes for the Detection of Cholinesterase Activity

Abstract

Acetylcholinesterase is one of the most widely used and studied enzymes. Not only does this enzyme regulate neurotransmission (and thus play a key role in neurodegenerative processes) but it is also a prime target for pest control agents and warfare agents. Above all, due to its particularly high turnover rate, acetylcholinesterase is among the most efficient reporter enzymes yet described (for use as enzymatic tracer in immunoassays, for instance). However, its activity is detected through a colorimetric reagent, the Ellman reagent, which displays low detection limits and is often subject to background perturbations. In the course of our search for a more sensitive detection assay, we describe here a first-generation 1,2-dioxetane chemiluminescent probe, based on chemically induced electron exchange luminescence, which has an approximately 10 times lower detection limit than the Ellman colorimetric assay (2.5 x 10(-19) mol for Electrophorus electricus AChE in its tetrameric form).

In situ induced metal-enhanced fluorescence: A new strategy for biosensing the total acetylcholinesterase activity in sub-microliter human whole blood

“…Several quantification techniques on AChE (Holas et al, 2012;Miao et al, 2010;Sabelle et al, 2002;Feng et al, 2007;Wang et al, 2009;Peng et al, 2009;Golub et al, 2013;Liao et al, 2013aLiao et al, , 2013bZhang et al, 2013;Garai-Ibabe et al, 2014) have been developed, each of which is suitable for different purposes and can be improved on user demand. The existing methods can solve problems in AChE activity determination.…”

Screening of acetylcholinesterase inhibitors by CE after enzymatic reaction at capillary inlet

“…In this method a yellow anion derived from the dithiobisnitrobenzoate (Ellman's reagent) when it reacts with TCh is monitored at 412 nm. Chemiluminiscent assays based on coupled enzymatic reactions have also been developed for the evaluation of AChE inhibitors [3,8,9].…”

Synthesis and structure–activity relationships of indole and benzimidazole piperazines as histamine H4 receptor antagonists

“…Commercially available 5-halo-2-indolecarboxylic acids were coupled with N-methylpiperazine using N,N 0 -carbonyldiimidazole (CDI) as coupling agent to form compound 1 and 2. 12 Halide exchange of compound 2 with potassium iodide and copper iodide in hexamethylphosphoric triamide (HMPA) 13,14 resulted in the iodo analogue 3. Using the same procedure as for the synthesis of 1 and 2, 5-chloro-2-indolecarboxylic acid was coupled to a range of amines to give analogues 4-9 as indicated in Scheme 2.…”