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.
Department of Molecular Oncology, H. Lee Moffitt Cancer Center and Research Institute, 12902 Magnolia Drive, Tampa, FL 33612, United States.
Liwei
L
Chen
Daniele
D
Pernazza
Latanya M
LM
Scott
Harshani R
HR
Lawrence
Yuan
Y
Ren
Yunting
Y
Luo
Xin
X
Wu
Shen-Shu
SS
Sung
Wayne C
WC
Guida
Said M
SM
Sebti
Nicholas J
NJ
Lawrence
Jie
J
Wu
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