Hit Validation Drug Discovery
Is Your Hit on Target?
How to Confidently Validate Drug Discovery Hits
High-throughput screening (HTS) is a powerful tool for identifying chemical starting points in drug discovery. But not all hits are created equal. Many compounds initially identified in screening fail to engage their intended targets in real-world conditions—leading to wasted resources and stalled projects.
The Challenge: Why Hit Confirmation Matters
Identifying a hit is just the beginning. Without rigorous hit confirmation, researchers risk advancing compounds that lack real target engagement, exhibit off-target effects, or fail due to poor cellular permeability. Traditional biochemical and biophysical assays, provide valuable insights but often fall short in reflecting physiological conditions due to tagging or labelling.
False positives can arise from various factors, including compound aggregation, non-specific binding, and assay interference. Advancing such misleading hits into later stages of drug discovery increases costs and time investment while also diverting resources from truly promising compounds. Furthermore, traditional target engagement assays may fail to capture complex cellular interactions, missing critical insights that could determine a compound’s success or failure. This is why robust target engagement methods, such as Pelago Bioscience’s CETSA®(Cellular Thermal Shift Assay), are crucial for confirming a compound’s activity in a physiologically relevant context.
CETSA: A Smarter Approach to Hit Confirmation
Our hit confirmation service offers a direct and reliable way to confirm target engagement in living cells—eliminating false positives and increasing confidence in hit validation.
Key advantages of CETSA:
✔ Physiologically relevant: Confirms target engagement in native cellular environments.
✔ Reduces false positives: Provides direct evidence of binding, minimizing misleading results.
✔ Accelerates hit-to-lead transition: Enables informed decision-making, prioritizing viable compounds.
Leading pharmaceutical companies have already adopted CETSA-based hit confirmation to streamline drug discovery efforts. For example, AstraZeneca successfully used CETSA to validate kinase inhibitors, reducing false positives and expediting lead optimization.
Want to See CETSA in Action?
At Pelago Bioscience, we specialize in CETSA-powered hit confirmation, helping researchers confidently advance their most promising compounds.
Our latest application note demonstrates how CETSA enhances hit validation, lead optimization, and target engagement analysis:
- Primary Screening: Learn how CETSA enabled AstraZeneca to screen 0.5 million compounds against CRAF, identifying known and novel inhibitors while minimizing false positives.
- Hit Confirmation: Discover how CETSA validated potent PARP1 binders by distinguishing active inhibitors from “silent binders,” offering a unique advantage over traditional assays.
- Potency ranking and SAR Analysis: See how CETSA was applied to a BRAF inhibitor screen, yielding highly selective hits and facilitating structure-activity relationship (SAR) analysis.
Integrating CETSA into your drug discovery workflow can accelerate the transition from hit identification to lead optimization, improving success rates and reducing costs.
References
- Jafari et al. Nature Protocols 2014
- Rowlands et al. SLAS Discovery 2023
- Shaw et al. SLAS Discovery 2018
