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Organic & Biomolecular Chemistry
Page 5 of 5
Journal Name
ARTICLE
Figure 4).18 Additionally, more potential BChE inhibitors can
be detected with the naked eye. As shown in Figure 4(C),
2
(
Silman, M. Harel, T. L. Rosenberry, J. L. Sussman, Chem. Biol.
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showed clearly different fluorescence intensities with and
without BChE under UV light illumination at 365 nm. The
inhibition efficiencies of potential BChE inhibitors could be
compared by determining the relative fluorescence intensities.
When the assay was performed with 100 nM of tacrine and
galantamine, BChE activity was completely inhibited by tacrine
3
4
(c) B. M. Liederer, R. T. Borchardt, J. Pharm. Sci., 2006, 95
1177.
,
and
with galantamine showed weak fluorescence intensity. This
result demonstrates that assay based on could be used not
1 showed no change in fluorescence intensity, whereas 1
5
6
7
V. P. Chen, Y. Gau, L. Geng, R. J. Parks, Y. P. Pang, S. Brimijoin,
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1
2
002, 9, 88.
only for real-time monitoring of BChE activity, but also for
high-throughput screening of BChE inhibitor candidates,
enabled by the noticeable visual change in fluorescence
intensity.
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,
1
4
. Conclusions
8
9
A choline substrate labeled with 4,5-dimethoxyphthalimide
7
,
was designed and synthesized for the purpose of selective and
direct assay of BChE. Compound was hydrolyzed by BChE,
1
1
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and its fluorescence was quenched by an intramolecular PET
process. Based on this fluorescence “turn-off” phenomenon, a
facile real-time and continuous fluorometric assay for BChE
has been developed. Notably, it was selectively hydrolyzed by
BChE even in the presence of excess AChE, because of which, it
can be used for the selective assay of BChE. In view of the
presence of excess AChE over BChE in vivo, it can be utilized
for selectively monitoring in-vivo BChE activity. Furthermore,
we have confirmed that this assay system could be used for
the screening of potential BChE inhibitors. Given the nature of
2
013, 85, 308; (e) J. Qian, X. Yang, L. Jiang, C. Zhu, H. Mao, K.
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,
2
5
its simplicity, selectivity, and assay speed,
1 can be used in the
1
1
high-throughput screening of selective BChE inhibitors and
relevant drug discovery.
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Acknowledgements
(
c) D. C. Vellom, Z. Radic, Y. Li, N. A. Pickering, S. Camp, P.
This research was a part of the project titled ‘Smart bio
sensing technology for managing distribution and safety about
fishery products and processed fishery products’, funded by
the Ministry of Oceans and Fisheries, Korea.
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