Identification of Novel Mast Cell Activators Using Cell-Based High-Throughput Screening

• Published in: SLAS discovery Vol. 24; no. 6; pp. 628 - 640
• Main Authors: Choi, Hae Woong, Chan, Cliburn, Shterev, Ivo D., Lynch, Heather E., Robinette, Taylor J., Johnson-Weaver, Brandi T., Shi, Jianling, Sempowski, Gregory D., Kim, So Young, Dickson, John K., Gooden, David M., Abraham, Soman N., Staats, Herman F.
• Format: Journal Article
• Language: English
• Published: Los Angeles, CA SAGE Publications 01.07.2019
• Subjects: Animals; Arachidonic Acid - metabolism; Biomarkers; Cell Degranulation - drug effects; Cell Line; Cell Survival - drug effects; Cytokines - metabolism; Dose-Response Relationship, Drug; Drug Discovery - methods; High-Throughput Screening Assays - methods; High-Throughput Screening Assays - standards; Humans; Mast Cells - drug effects; Mast Cells - immunology; Mast Cells - metabolism; Mice; Quality Control; Small Molecule Libraries; mast cell; degranulation; mast cell activator; high-throughput screening
• ISSN: 2472-5552; 2472-5560
• DOI: 10.1177/2472555219834699

Mast cells (MCs) are known to regulate innate and adaptive immunity. MC activators have recently been described as safe and effective vaccine adjuvants. Many currently known MC activators are inadequate for in vivo applications, however, and research on identifying novel MC activators is limited. In this study, we identified novel MC activators by using high-throughput screening (HTS) assays using approximately 55,000 small molecules. Data sets obtained by the primary HTS assays were statistically evaluated using quality control rules and the B-score calculation, and compounds with B-scores of >3.0 were chosen as mast cell activators (hits). These hits were re-evaluated with secondary and tertiary HTS assays, followed by further statistical analysis. From these hits, we selected 15 compounds that caused degranulation in murine and human MCs, with potential for flexible chemical modification for further study. Among these 15 compounds, ST101036, ST029248, and ST026567 exhibited higher degranulation potency than other hit compounds in both human and mouse MCs. In addition, the 15 compounds identified promote de novo synthesis of cytokines and induce the release of eicosanoids from human and mouse MCs. HTS enabled us to identify small-molecule MC activators with unique properties that may be useful as vaccine adjuvants.