Permeability Enhancer Incorporated Oral Nanosuspension of ARV-825 PROTAC for Glioblastoma Treatment

Authors: Patel H, Yadav A, Dukhande V, Patel K
Publication: Journal of Pharmaceutical Investigation
Keywords: drug delivery, drug development, pharmaceutics, phase IV trials, therapeutics
Software: GastroPlus®

Abstract

Purpose
Glioblastoma(GBM) is an aggressive brain tumor with dismal prognosis, necessitating innovative therapeutic strategies. Proteolysis-targeting chimeras(PROTACs), such as ARV-825, hetero-bifunctional degrader of BRD4 offer targeted protein degradation but suffer from poor aqueous solubility, high molecular weight, and limited permeability, resulting in low oral bioavailability. This study aimed to develop nanosuspension formulation (A-NS) of ARV-825 to enhance its solubility, stability, and oral absorption for GBM therapy.

Methods
A-NS was prepared via dual centrifugation nano-milling using sodium lauryl sulfate, polyvinyl pyrrolidone, and SNAC as co-stabilizers and permeation enhancer. The formulation was characterized for particle size, zeta potential, and in-vitro dissolution. in-silico physiologically based pharmacokinetic(PBPK) simulations were used to predict oral bioavailability. ARV-825 and A-NS were evaluated for cytotoxicity and 3D spheroid disruption in TMZ-sensitive and resistant GBM cell lines. RNA sequencing was conducted to elucidate downstream genes and pathway alterations.

Results
A-NS achieved nanoscale size(~204 nm), enhanced solubility, and improved dissolution. PBPK modeling predicted substantial increase in oral bioavailability(~87%) versus parent ARV-825(~6%). In-vitro studies confirmed improved cytotoxicity in both TMZ-sensitive and resistant GBM cells. Western blot analysis revealed effective BRD4 degradation, RNA sequencing identified enrichment of apoptosis-related pathways and suppression of oncogenes. 3-D spheroid assay demonstrated significant spheroid disruption in TMZ-sensitive and resistant GBM cells by ~ 5 and ~ 3 folds respectively.

Conclusion
The developed A-NS formulation of ARV-825 overcomes key biopharmaceutical limitations, enabling effective oral delivery of PROTACs. Overall, oral delivery system offers promising avenue for GBM treatment, with notable efficacy in temozolomide-resistant cells. Further in-vivo validation is essential to establish its translational potential.

By Himaxi Patel, Anjali Yadav, Vikas Dukhande & Ketankumar Patel