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.
Pfizer Research Technology Center, 620 Memorial Drive, Cambridge, MA 02139, USA. haile.tecle@pfizer.com
Haile
H
Tecle
Frederic
F
Feru
Hu
H
Liu
Cyrille
C
Kuhn
Glen
G
Rennie
Mark
M
Morris
Jiangxing
J
Shao
Alan C
AC
Cheng
Diana
D
Gikunju
Juan
J
Miret
Rocco
R
Coli
Simon Hualin
SH
Xi
Susan L
SL
Clugston
Simon
S
Low
Steven
S
Kazmirski
Yuan-Hua
YH
Ding
Qing
Q
Cao
Theresa L
TL
Johnson
Gayatri D
GD
Deshmukh
Jonathan P
JP
DiNitto
Joe C
JC
Wu
Jessie M
JM
English
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