High-throughput identification of inhibitors of human mitochondrial peptide deformylase.
The human mitochondrial peptide deformylase (HsPDF) provides a potential new target for broadly acting antiproliferative agents. To identify novel nonpeptidomimetic and nonhydroxamic acid-based inhibitors of HsPDF, the authors have developed a high-throughput screening (HTS) strategy using a fluorescence polarization (FP)-based binding assay as the primary assay for screening chemical libraries, followed by an enzymatic-based assay to confirm hits, prior to characterization of their antiproliferative activity against established tumor cell lines. The authors present the results and performance of the established strategy tested in a pilot screen of 2880 compounds and the identification of the 1st inhibitors. Two common scaffolds were identified within the hits. Furthermore, cytotoxicity studies revealed that most of the confirmed hits have antiproliferative activity. These findings demonstrate that the designed strategy can identify novel functional inhibitors and provide a powerful alternative to the use of functional assays in HTS and support the hypothesis that HsPDF inhibitors may constitute a new class of antiproliferative agent.
Antczak
Christophe
C
High Throughput Screening Core Facility, Memorial Sloan-Kettering Cancer Center, New York, New York 10021, USA.
Shum
David
D
Escobar
Sindy
S
Bassit
Bhramdeo
B
Kim
Earl
E
Seshan
Venkatraman E
VE
Wu
Nian
N
Yang
Guangli
G
Ouerfelli
Ouathek
O
Li
Yue-Ming
YM
Scheinberg
David A
DA
Djaballah
Hakim
H
D000581:Amidohydrolases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000378:metabolism*; D018922:HL-60 Cells; D006801:Humans; D019169:Jurkat Cells; D020014:K562 Cells; D008928:Mitochondria / Q000187:drug effects / Q000201:enzymology*; D021241:Spectrometry, Mass, Electrospray Ionization
D004791:Enzyme Inhibitors; D000581:Amidohydrolases; C084507:peptide deformylase