The identification of GPR3 inverse agonist AF64394; the first small molecule inhibitor of GPR3 receptor function.

The identification of the novel and selective GPR3 inverse agonist AF64394, the first small molecule inhibitor of GPR3 receptor function, is described. Structure activity relationships and syntheses based around AF64394 are reported.

Profiling of multiple signal pathway activities by multiplexing antibody and GFP-based translocation assays.

Multiplexing of GFP based and immunofluorescence translocation assays enables easy acquisition of multiple readouts from the same cell in a single assay run. Immunofluorescence assays monitor translocation, phosphorylation, and up/down regulation of endogenous proteins. GFP-based assays monitor translocation of stably expressed GFP-fusion proteins.

High content translocation assays for pathway profiling.

This chapter describes the design and development of cell-based assays, in which quantitation of the intracellular translocation of a target protein--rather than binding or catalytic activity--provides the primary assay readout. These are inherently high content assays, and they provide feedback on cellular response at the systems level, rather than data on activities of individual, purified molecules. Multiple protein translocation assays can be used to profile cellular signaling pathways and they can play a key role in determination of mechanism of action for novel classes of compounds with therapeutic potential.

Identification of RAS-mitogen-activated protein kinase signaling pathway modulators in an ERF1 redistribution screen.

The RAS-mitogen-activated protein kinase (MAPK) signaling pathway has a central role in regulating the proliferation and survival of both normal and tumor cells. This pathway has been 1 focus area for the development of anticancer drugs, resulting in several compounds, primarily kinase inhibitors, in clinical testing. The authors have undertaken a cell-based, high-throughput screen using a novel ERF1 Redistribution assay to identify compounds that modulate the signaling pathway.

Non-anti-coagulant heparins: a promising approach for prevention of tumor metastasis (review).

The metastatic spread of solid tumors is responsible for most cancer-related deaths, and unfortunately no specific anti-metastatic therapy exists. Heparin is widely used in the clinic today as an anti-coagulant therapy for cancer patients at risk for venous thrombo-embolism. Recent clinical trials with low molecular weight heparin and meta-analyses of earlier clinical trials with unfractionated heparin indicate that heparin has an anti-metastatic activity.