Exploring the Therapeutic Potential of Cannflavins and Stilbenes: From Pharmaceutical  Computational Modeling to Formulation Development
Creators: O'Croinin, Conor J.
Publisher: name: University of Alberta Library
Year: 2024

<p>Background: Cannabis sativa has been demonstrated to have numerous physiological effects. The plant contains hundreds of chemical constituents including well studied cannabinoids like cannabidiol and delta-9-tetrahydrocannabinol as well as poorly characterized compounds like polyphenols. Polyphenols in cannabis include stilbenes and flavonoids that are largely understudied for their therapeutic effects. Stilbenes present in cannabis include dihydrostilbenoids and some specific flavonoids are named cannflavins. These polyphenols have been characterized to various degrees, and further characterization can be done using predictive in silico pharmaceutical modeling and pharmacodynamic screening assays. Pharmaceutical investigations are needed to create prototype formulations specific to these compounds, with several potential routes of administration being considered for polyphenol delivery. Models have been established to determine the effectiveness of the penetrance, dissolution, lipolysis, and lymphatic uptake for the active flavonoid ingredient in a formulation that can be applied to novel formulation development and optimization.  Methods: In silico predictive modeling was performed using ADMET PredictorTM to obtain predictive data on the metabolism, permeability, physicochemical properties, and pharmacokinetics of the polyphenols. A method of analysis was developed and validated for the quantification of cannflavins A, B, and C using liquid chromatography-mass spectrometry (LCMS). Prototype formulations were developed for topical, transdermal patch, oral dissolving film, and oral capsule delivery. These formulations were evaluated for penetrance through synthetic and ex vivo porcine membranes using Franz diffusion cells and dissolution was assessed using a USP II dissolution apparatus. Colorimetric and fluorescent enzymatic screening assays were performed to determine potential novel pharmacological targets of the polyphenols. <br>
 Results: In silico data highlighted the pharmacokinetic properties of the polyphenols, potential activity against HIV-1 integrase and breast cancer resistance protein, while also predicting metabolic interactions with CYP and UGT enzymes in the body and revealed potential drug-drug interactions. We have validated the first method of analysis for the LCMS analysis of cannflavins A, B, and C simultaneously. The method is isocratic, simple, robust, and sensitive and permits the quantitative analysis using electrospray LCMS. Prototype formulations were successfully developed for topical, transdermal patch, oral dissolving film, and oral capsule routes of administration. The most effective forms of cannflavin A delivery developed were topical and oral capsule formulations. Oral capsules demonstrated effective dissolution with the addition of lipid based excipients to the formulation to take advantage of lipolysis and lymphatic uptake. Pharmacological screening assays further determined potential novel antiviral, anti-inflammatory, analgesic, and neuroprotective effects of the polyphenols.  Conclusion: In conclusion, we have demonstrated novel findings using in silico modeling and in vitro screening assays regarding the potential therapeutic properties and targets of cannabis-derived stilbenes and flavonoids. We have created and validated a novel method of analysis using LCMS to quantify cannflavins. Prototype pharmaceutical formulations for the delivery of cannflavins through multiple dosage forms have been created and proven effective at releasing cannflavins across synthetic and ex vivo membranes. </p>