Case Story: CETSA® in Translational Drug Discovery
Bridging In Vitro Observations
with In Vivo Outcomes
The transition from simplified in vitro systems to physiologically complex in vivo models presents a persistent challenge in drug discovery. Conventional in vitro assays often fail to replicate the cellular heterogeneity, tissue-specific signaling, and systemic pharmacokinetics observed in living organisms. As a result, pharmacological activity measured in vitro may not translate predictably across tissues or species.
To improve translational accuracy, it is essential to select in vitro models that reflect relevant disease biology and target expression. Thoughtfully designed cellular systems can enhance predictive value and inform key decisions in early development.
Scientific Challenge
A client advancing a novel therapeutic target had established strong structure-activity relationships and favorable potency in vitro. However, critical translational questions remained: Does the compound engage its molecular target within a biologically relevant, complex environment? Can inhaled administration achieve therapeutic concentrations in lung tissue?
To address these questions. CETSA® was implemented in disease-relevant cell lines and ex vivo lung tissue from in vivo dosed mice.
CETSA-Driven Translational Workflow
The experimental strategy followed a staged design to generate functional and pharmacodynamic insight:
1. Cell-Based CETSA Assay: Target engagement was first confirmed in a relevant cell model. This demonstrated that the compound retained activity in a cellular context, accounting for intracellular dynamics and potential metabolic effects.
2. Ex Vivo CETSA in In Vivo-Dosed Lung Tissue: Following inhalational dosing in a murine model, lung tissue was collected and analyzed using CETSA. This provided direct evidence of target engagement at the site of action and validated the intended delivery route.
3. Pharmacodynamic Time-Course Analysis: Tissues were sampled at multiple time points post-dose to assess the dynamics of target engagement over time. These data informed the pharmacokinetic-pharmacodynamic relationship and supported dose scheduling decisions.
Translational Outcome: The CETSA-guided approach provided the client with high-confidence data:
- Target engagement was verified in both in vitro and in vivo contexts
- Inhalation was confirmed as an effective delivery route for the compound
- Time-resolved analysis revealed consistent pharmacodynamic behavior
These findings validated the therapeutic hypothesis and supported progression into advanced development.
STRATEGIC INSIGHTS: This case study illustrates how CETSA can serve as a robust translational tool. Confirming molecular target engagement across preclinical systems reduces uncertainty and increases the likelihood of successful translation from bench to clinic
Webinar: How CETSA® Enhances Translational Studies – A Sitryx Case Study
In this on-demand webinar, Jon Ellery, Director of Biology at Sitryx, shares how CETSA technology helped Sitryx assess target engagement and improve translational confidence in small molecule drug discovery.
Is your compound actually therapeutic?
Learn how CETSA confirms target engagement across cell lines, animal models, and human primary cells—providing physiologically relevant insights that streamline drug discovery.
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