Recently, I had the opportunity to organize a roundtable at the Eighth Western Canadian Medicinal Chemistry Workshop hosted by my university.

Given the diverse expertise of our invited speakers—radiochemistry, natural products, synthetic methodology, and host-pathogen interactions—for the session, I organized a set of questions that, in my experience, are essential to address when evaluating biological experiments.

Below, I present these questions alongside applied examples:

Questions 1–3 aim to analyze:

First,  How do you unequivocally validate that a specific biological target, cell-surface receptor, or chemical signal is the true driver of an observed response?

Let’s examine how these questions apply to my own research:

During my PhD, I investigated α-synuclein aggregation. α-Synuclein is a cytosolic protein that misfolds into oligomers, protofibrils, mature fibrils, and amorphous aggregates that drive progressive neuronal atrophy.

In an in vitro assay involving α-synuclein overexpression, distinguishing the exact aggregate species—whether oligomers, protofibrils, or amorphous aggregates—is often challenging. Consequently, attributing an observed inhibitory signal directly to the suppression of protofibrils or amorphous fibrils remains difficult.

Second, what is the most common failure you have faced when transitioning from a
promising an analytical method-characterization and how did you adapt your methods when this occurred?

e.g. Transitioning an α-synuclein aggregation inhibitor into a functional molecular probe for cellular assays typically requires structural modification. For instance, incorporation of water-soluble tags within click-compatible tags to facilitate fluorescent visualization and enable target protein pull-down.

And third: what specific parameter, chemical structure, or thermodynamic parameter serves as your primary green light to move from an initial result to an advanced test?

e.g. How many experiments are required to validate a lead compound? 

As a conclusion I would like to appoint that: