Abstract
Purpose :
We have generated and tested a synthetic library of novel cannabinoid-based molecules with the potential to treat dry eye disease (DED). Here, we present a rigorous in vitro screening platform based on activity or inhibition of pathways relevant to cannabinoid biology in the context of ocular pathologies.
Methods :
A compound library with approximately 100 novel cannabinoid-based molecules were chemically synthesized and interrogated for their biological activity. An ocular screening platform was developed using immortalized human ocular cell lines (conjunctival epithelial, corneal epithelial/endothelial, keratocytes, retinal endothelial), stable HEK cell lines engineered to measure activation/inhibition of specific cannabinoid receptors (CB1, CB2, GPR55, TRPV1), as well as transient transfection of various elements to measure specific biological activities such as cytotoxicity, wound healing, cellular antioxidant/hypoxic activity and induction of Nrf2, PPARa/g NF-kB signaling.
Results :
To evaluate various biological parameters, activity/inhibition was evaluated via fold induction over the control basal level. Using a 2X threshold of relevant biological activity, 38 compounds were selected for additional testing to determine EC50/IC50 in key pathways related to DED such as inhibition of NF-kB, activation of CB2 signaling, inhibition of ROS/HIF-1 activity and inhibition of TRPV-1 signaling. In addition to biological testing, these 38 compounds were additionally interrogated for in silico attributes such as synthesis complexity, lipophilicity, water solubility and application of drug-likeness rules. Ultimately, cannabinerol, a trace constituent of cannabis, and two non-electrophilic indolic adducts of cannabidiolquinone and cannabigeroquinone were selected for further pharmaceutical development.
Conclusions :
Here we report data in support of a novel DED therapeutic, using a robust in vitro screening platform to efficiently interrogate ocular-specific biology. Coupled with in silico analysis, this approach may rapidly advance pharmaceutical development of synthetic cannabinoid-based therapeutics with the potential to treat ocular pathologies like DED or chronic ocular pain.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.