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Identification of Novel BACE-1 Inhibitors through an Advanced Structure-Based Virtual Screening Protocol
Puneet Kacker, PhD Student, Italian Institute of Technolog, Genovay
BACE-1 is considered a promising therapeutic target for the treatment of Alzheimer’s disease. Computational studies performed on BACE-1 must address two main issues: (1) the protonation state of the catalytic dyad varies in the presence of different interacting molecules, and (2) the ligand binding site is extremely flexible. Here, a virtual ligand screening protocol that takes simultaneously into account the contributions of dyad protonation and binding pocket rearrangements to binding affinity is proposed. Combining quantum mechanical calculations, molecular dynamics simulations, and docking studies, an optimal conformational ensemble encompassing 10 conformers and 4 protonation states that captured the overall enzyme plasticity was identified. Thanks to the application of the reported protocol to a library of over 70,000 commercially available compounds, it was possible to prioritize a series of 13 analogues for in vitro testing. Among them, the most active compound (63.2 µM IC50) was selected for a hit improvement program.
Quantification of cytokines on a Multi-Mode Microplate Detection Platform using Alpha Technology
Carlene Whitehead, European Marketing Communications Specialist, Molecular Devices
Inflammation is accompanied by increased endothelial chemokine production and adhesion molecule expression, which may result in an extensive neutrophil infiltration. As such, the search for novel anti-inflammatory substances able to downregulate these parameters, as well as the tissue damage, holds therapeutic promise. Here we describe how Alpha Technology, a bead-based, homogeneous assay for studying molecular interactions in a microplate format, has been used to detect anti-inflammatory metabolites from cyanobacteria in human endothelial cell-based in vitro assays. We show the quantification of cytokines (TNF-alpha) down to picogram levels with Molecular Devices' SpectraMax® Paradigm® Modular Multi-Mode Reader. Alpha Technology can be performed on the SpectraMax Paradigm Reader in various plate formats ranging from 96-well to 1536-well.
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