The design, synthesis and potential utility of fluorescence probes that target DFG-out conformation of p38alpha for high throughput screening binding assay.
The design, synthesis and utility of fluorescence probes that bind to the DFG-out conformation of p38alpha kinase are described. Probes that demonstrate good affinity for p38alpha, have been identified and one of the probes, PF-04438255, has been successfully used in an high throughput screening (HTS) assay to identify two novel non-classical p38alpha inhibitors. In addition, a cascade activity assay was utilized to validate the selective binding of these non-classical kinase inhibitors to the unactive form of the enzyme.
Tecle
Haile
H
Pfizer Research Technology Center, 620 Memorial Drive, Cambridge, MA 02139, USA. haile.tecle@pfizer.com
Feru
Frederic
F
Liu
Hu
H
Kuhn
Cyrille
C
Rennie
Glen
G
Morris
Mark
M
Shao
Jiangxing
J
Cheng
Alan C
AC
Gikunju
Diana
D
Miret
Juan
J
Coli
Rocco
R
Xi
Simon Hualin
SH
Clugston
Susan L
SL
Low
Simon
S
Kazmirski
Steven
S
Ding
Yuan-Hua
YH
Cao
Qing
Q
Johnson
Theresa L
TL
Deshmukh
Gayatri D
GD
DiNitto
Jonathan P
JP
Wu
Joe C
JC
English
Jessie M
JM
D001665:Binding Sites; D003198:Computer Simulation; D018360:Crystallography, X-Ray; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D057166:High-Throughput Screening Assays; D007700:Kinetics; D048308:Mitogen-Activated Protein Kinase 14 / Q000737:chemistry / Q000378:metabolism*; D009281:Naphthalenes / Q000737:chemistry / Q000494:pharmacology; D047428:Protein Kinase Inhibitors / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D011720:Pyrazoles / Q000737:chemistry / Q000494:pharmacology; D013329:Structure-Activity Relationship; D010671:Phenylurea Compounds
D005456:Fluorescent Dyes; D048308:Mitogen-Activated Protein Kinase 14; D009281:Naphthalenes; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; C452139:doramapimod; D010671:Phenylurea Compounds