Large-scale Protein Expression
Having sufficient quantities of reagents is a basic prerequisite to successful high-throughput screening (HTS). Often, supplies of target proteins are limited because most academic labs do not have the capability to produce the necessary amount of proteins. We plan to collaborate with the X-ray Crystallography Core Facility to establish large-scale protein expression capability to improve the protein supply bottleneck.
Development of In Silico ADME Prediction and In Vitro ADME Testing
One of the major obstacles in drug discovery and development is the high failure rate of compounds during clinical trials. The major causes of failure could result from poor absorption, distribution, metabolization and excretion (ADME) properties. With the advancement of computational power and methodologies, it is now possible to perform in silico calculation of a number of important ADME properties including aqueous solubility, log P octane/water, Caco-2 cell permeability and human serum albumin binding. By combining the in silico calculation with in vitro ADME testing to confirm the predicted values, we can more accurately target selection of lead compounds for further development. The Biomedical Informatics Core will purchase ADME prediction software and make it available through City of Hope’s intranet. The HTS Core will establish in vitro ADME assays, including Biomimetic Artificial Membrane Permeation Assay (BAMPA), Transil Human Serum Albumin (HSA) Binding Assay and Cytochrome P450 Isoenzymes Assay, to obtain experimental data on key ADME properties for compounds of interest.
Interaction between Cores
The following image shows interactions between the High Throughput Screening (green), Biomedical Informatics (yellow) and Synthetic and Biopolymer Chemistry (purple).
Experimental screenings performed in the High Throughput Screening (HTS) Core complement virtual screenings carried out by the Biomedical Informatics Core toward identifying “hits” for biomolecular targets of interest.
Confirmed hits, also known as leads, identified through screenings are used as starting points for chemists in the Synthetic and Biopolymer Chemistry Core for designing focused libraries. Molecular modeling expertise in the Biomedical Informatics Core aids the design process.
Chemists synthesize the focused libraries accordingly, which in turn are tested by HTS.
Structure-activity relationship analysis is then carried out to evaluate newly synthesized compounds using appropriate in vitro or cell-based assays.
Iterative cycles of interactions of the three cores will speed up the “lead optimization” process and move the project forward.