Inhibition of cellular Shp2 activity by a methyl ester analog of SPI-112.
The protein tyrosine phosphatase (PTP) Shp2 (PTPN11) is an attractive target for anticancer drug discovery because it mediates growth factor signaling and its gain-of-function mutants are causally linked to leukemias. We previously synthesized SPI-112 from a lead compound of Shp2 inhibitor, NSC-117199. In this study, we demonstrated that SPI-112 bound to Shp2 by surface plasmon resonance (SPR) and displayed competitive inhibitor kinetics to Shp2. Like some other compounds in the PTP inhibitor discovery efforts, SPI-112 was not cell permeable, precluding its use in biological studies. To overcome the cell permeation issue, we prepared a methyl ester SPI-112 analog (SPI-112Me) that is predicted to be hydrolyzed to SPI-112 upon entry into cells. Fluorescence uptake assay and confocal imaging suggested that SPI-112Me was taken up by cells. Incubation of cells with SPI-112Me inhibited epidermal growth factor (EGF)-stimulated Shp2 PTP activity and Shp2-mediated paxillin dephosphorylation, Erk1/2 activation, and cell migration. SPI-112Me treatment also inhibited Erk1/2 activation by a Gab1-Shp2 chimera. Treatment of Shp2(E76K) mutant-transformed TF-1 myeloid cells with SPI-112Me resulted in inhibition of Shp2(E76K)-dependent cell survival, which is associated with inhibition of Shp2(E76K) PTP activity, Shp2(E76K)-induced Erk1/2 activation, and Bcl-XL expression. Furthermore, SPI-112Me enhanced interferon-gamma (IFN-gamma)-stimulated STAT1 tyrosine phosphorylation, ISRE-luciferase reporter activity, p21 expression, and the anti-proliferative effect. Thus, the SPI-112 methyl ester analog was able to inhibit cellular Shp2 PTP activity.
Chen
Liwei
L
Department of Molecular Oncology, H. Lee Moffitt Cancer Center and Research Institute, 12902 Magnolia Drive, Tampa, FL 33612, United States.
Pernazza
Daniele
D
Scott
Latanya M
LM
Lawrence
Harshani R
HR
Ren
Yuan
Y
Luo
Yunting
Y
Wu
Xin
X
Sung
Shen-Shu
SS
Guida
Wayne C
WC
Sebti
Said M
SM
Lawrence
Nicholas J
NJ
Wu
Jie
J
D001667:Binding, Competitive; D002461:Cell Line, Transformed; D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D019073:HT29 Cells; D006801:Humans; D007211:Indoles / Q000737:chemistry* / Q000494:pharmacology*; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11 / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*; D013449:Sulfonamides / Q000494:pharmacology*; D013451:Sulfonic Acids / Q000737:chemistry* / Q000494:pharmacology*
C531229:3-(2-(2-nitrophenyl)hydrazono)-2-oxoindoline-5-sulfonic acid; C551405:3-(2-(5-(N-(4-fluorobenzyl)sulfamoyl)-2-oxoindolin-3-ylidene)hydrazinyl)benzoic acid methyl ester; D004791:Enzyme Inhibitors; D007211:Indoles; D013449:Sulfonamides; D013451:Sulfonic Acids; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11