In:
ChemBioChem, Wiley, Vol. 14, No. 13 ( 2013-09-02), p. 1611-1619
Abstract:
Despite the recognized importance of membrane proteins as pharmaceutical targets, the reliable identification of fragment hits that are able to bind these proteins is still a major challenge. Among different 19 F NMR spectroscopic methods, n ‐fluorine atoms for biochemical screening ( n ‐FABS) is a highly sensitive technique that has been used efficiently for fragment screening, but its application for membrane enzymes has not been reported yet. Herein, we present the first successful application of n ‐FABS to the discovery of novel fragment hits, targeting the membrane‐bound enzyme fatty acid amide hydrolase (FAAH), using a library of fluorinated fragments generated based on the different local environment of fluorine concept. The use of the recombinant fusion protein MBP‐FAAH and the design of compound 11 as a suitable novel fluorinated substrate analogue allowed n ‐FABS screening to be efficiently performed using a very small amount of enzyme. Notably, we have identified 19 novel fragment hits that inhibit FAAH with a median effective concentration (IC 50 ) in the low m M –μ M range. To the best of our knowledge, these results represent the first application of a 19 F NMR fragment‐based functional assay to a membrane protein.
Type of Medium:
Online Resource
ISSN:
1439-4227
,
1439-7633
DOI:
10.1002/cbic.201300347
Language:
English
Publisher:
Wiley
Publication Date:
2013
detail.hit.zdb_id:
2019276-9
detail.hit.zdb_id:
2020469-3
SSG:
12
