Validation of surface plasmon resonance screening of a diverse chemical library for the discovery of protein tyrosine phosphatase 1b binders.
We investigated the suitability of surface plasmon resonance (SPR) for providing quantitative binding information from direct screening of a chemical library on protein tyrosine phosphatase 1b (PTP1B). The experimental design was established from simulations to detect binding with K(D) < 10⁻⁴ M. The 1120 compounds (cpds) were injected sequentially at concentrations [C(cpd)] of 0.5 or 10 μM over various target surfaces. An optimized evaluation procedure was applied. More than 90% of cpds showed no detectable signal in four screens. The 30 highest responders at C(cpd)=10 μM, of which 25 were selected in at least one of three screens at C(cpd)=0.5 μM, contained 22 promiscuous binders and 8 potential PTP1B-specific binders with K(D) ~10⁻⁵ M. Inhibition of PTP1B activity was assayed and confirmed for 6 of these, including sanguinarine, a known PTP1B inhibitor. C(cpd) dependence studies fully confirmed screening conclusions. The quantitative consistency of SPR data led us to propose a structure-activity relationship (SAR) model for developing selective PTP1B inhibitors based on the ranking of 10 arylbutylpiperidine analogs.
Biosensor Group, UMR 7242, CNRS-University of Strasbourg, 67412 Illkirch, France.
Gabrielle
G
Zeder-Lutz
Laurence
L
Choulier
Marie
M
Besse
Alexandra
A
Cousido-Siah
Francesc Xavier
FX
Ruiz Figueras
Bruno
B
Didier
Marie-Louise
ML
Jung
Alberto
A
Podjarny
Danièle
D
Altschuh
D000595:Amino Acid Sequence; D055808:Drug Discovery / Q000379:methods*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D020128:Inhibitory Concentration 50; D016366:Open Reading Frames; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D054852:Small Molecule Libraries; D020349:Surface Plasmon Resonance / Q000379:methods*
D004791:Enzyme Inhibitors; D054852:Small Molecule Libraries; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1