Objectives:
To develop a next-generation CBD analogue platform through rational scaffold engineering and targeted derivatization. The structural changes were designed to reduce CBD’s major liabilities of CYP inhibition, poor solubility, rapid metabolism, and hepatic concerns, while preserving the key molecular interactions responsible for its antiseizure activity. A central design principle was to achieve higher and more sustained therapeutic brain concentrations, enabling predictable pharmacokinetics and once-daily oral dosing.

Methods:
More than 50 CBD analogues were generated using a scaffold-engineering strategy guided by known metabolic hotspots and physicochemical liabilities of cannabidiol. Structural modifications were selected to improve solubility, metabolic stability, CYP interaction profiles, and off-target safety. Analogues were evaluated in vitro for solubility, microsomal stability, CYP inhibition, hepatocyte viability, and receptor-level safety screens. Lead candidates underwent in vivo plasma and brain pharmacokinetic studies in mice. Antiseizure efficacy was assessed following oral administration across three validated preclinical seizure models. Exploratory hepatic and tolerability assessments supported translational evaluation.

Results:
In vitro profiling showed that scaffold-engineered CBD analogues achieved substantial improvements in solubility, enhanced metabolic stability, and markedly reduced CYP inhibition across major isoforms, indicating a significantly lower drug–drug interaction risk. Exploratory safety assays demonstrated higher hepatocyte viability, cleaner hepatic biomarker profiles, and reduced off-target activity at cardiovascular, opioid, and cholinergic receptors, supporting a more favorable safety margin.
These in vitro gains translated into major in vivo pharmacokinetic advantages. Lead analogues produced multi-fold increases in CNS exposure (up to six-fold higher brain concentrations than CBD) and improved brain-to-plasma ratios, resulting in more sustained and therapeutically relevant brain levels. Importantly, PK profiles showed markedly lower inter-animal variability, yielding predictable exposure kinetics compatible with once-daily dosing.
Improved exposure and preserved mechanistic interactions produced greater antiseizure potency and efficacy across three validated preclinical seizure models, including the MES assay. The optimized analogues achieved robust oral efficacy at lower doses due to higher and longer-lasting therapeutic brain concentrations. Together, these improvements support a broader therapeutic index and validate the scaffold-engineering strategy as a viable route for creating drug-like cannabidiol analogues

Conclusions:
Rational synthetic derivatization and medicinal chemistry enabled the creation of next-generation CBD analogues that overcome major drug-like liabilities of cannabidiol while preserving the molecular features required for antiseizure activity. The resulting compounds show improved solubility, reduced CYP interactions, enhanced hepatic and off-target safety, and multi-fold increases in brain exposure with more predictable, once-daily pharmacokinetics. These advances produced superior oral antiseizure efficacy at lower doses and support this platform as a promising route for developing clinically viable cannabidiol-based therapeutics for refractory epilepsy.

Learning Objectives:

• Understand how scaffold-engineered CBD analogues improve solubility, metabolism, CYP and safety profiles, achieve higher and longer-lasting therapeutic brain levels, and support predictable once-daily oral dosing with strong anti-seizure efficacy