The Drug Discovery Seed Compounds Exploratory Unit aims to identify seed and lead compounds for drug development that are active against drug target molecules through high-throughput screening (HTS) of large compound libraries.
Assay design and development for target molecules and phenotypes are one of the most important tasks in HTS, and we propose and construct assay systems appropriate for drug targets. Semi-automation of assay systems for HTS and hit compound validation via molecular interaction analysis are also conducted in our unit. Furthermore, we can contribute to the selection of true hit compounds and the generation of lead candidate compounds through profiling assays.
To find seed compounds that match the drug discovery concept of research projects, we conduct HTS campaigns by constructing assay systems using appropriate materials and measurement indicators. We use a variety of assay systems based on the family and type of target molecules and cell-free or cell-based systems that are more relevant.
| Assay types | In vitro biochemical assay (Cell-free assay) |
Cell-based assay | |
|---|---|---|---|
| Target-oriented assay | Phenotypic assay | ||
| Targets | Enzymes, Receptors, Protein-protein interaction, etc. | GPCRs, Receptors, Ion channels, Nuclear hormone receptors, Protein-protein interactions, Signaling pathways, etc. | Morphology, localization, numbers, and intensities of cells, organelles, cellular structures, and molecules |
| Methods | Absorbance, Fluorescence Intensity, Fluorescence Polarization (FP), Fluorescence Resonance Energy Transfer (FRET), Time-Resolved FRET, Alpha Screen/AlphaLISA, Surface plasmon resonance (SPR), Microscale thermophoresis (MST), Differential Scanning Fluorimetry (DSF), Calorimetry | Reporter assay (Luciferase, SEAP etc.), Cell imaging (In cell FRET, Fluorescent proteins, Immunostaining), Yeast system, FLIPR/FDSS (Aequorin, Ca2+ sensor, other sensors), Split luciferase complementation, etc. | Cell imaging (Fluorescent proteins, Organella-specific staining, Immunostaining) |
We have various high-throughput dispensers, microplate imagers, high-end microplate readers, and other measurement systems, as shown in Figure 1. HTS with all assay methods shown in Table 1 can be performed in our unit. The large amounts of data generated by HTS are analyzed and managed via a screening database (Figure 2).
Click on the following image to enlarge
Click on the following image to enlarge
We evaluate hit compounds by biochemical assays and physical binding analysis using surface plasmon resonance (SPR) and microscale thermophoresis (MST) and use isothermal titration calorimetry (ITC) analysis to identify true hits. We also contribute to the profiling of hit compounds for the generation of lead candidate compounds.
When the target of a promising compound is unknown, target identification can be performed in collaboration with the Chemical Genomics Research Group/Molecular Ligand Target Research Team.
The business development office handles alliances for collaboration with companies, universities, and external research institutes. It is also involved in licensing intellectual property rights to companies in collaboration with RIKEN Innovation.
Mission of Translational Research Office (TRO), the competent team which consists of number of experts of translational research (TR) from industry, academic TR institutes, regulatory organizations, is to support projects establishing clinical development plan.
