Abstract
Intravitreal delivery of antibody-based therapeutics has revolutionized the treatment of intraocular vascular diseases involving the retina and choroid. Unfortunately, limited durability requires frequent retreatment placing an enormous burden on patients. We sought to solve this problem with a novel approach that uses an anchoring molecule characterized by two key molecular properties: (1) non-covalent binding to an antibody-based therapeutic, and (2) retention in the vitreous cavity. As an initial proof-of-principle, we chose an anchoring molecule composed of agarose microbeads functionalized with an Fc-binding domain. Bevacizumab was chosen as the antibody-based therapeutic. In vitro experiments demonstrated that bevacizumab was maximally bound to this anchoring molecule within 1 h, and was competitively released upon exposure to either polyclonal human (p < 0.0001) or rat (p = 0.0017) immunoglobulins. In silico modeling predicted prolonged intravitreal retention of an antibody-based therapeutic in the presence of this anchoring molecule, which was confirmed by in vivo experiments with this initial anchoring molecule in rats. This anchoring molecule increased the intraocular half-life of bevacizumab from 5.8 days to over 18 days and maintained therapeutic concentrations for over 80 days. Despite showing no evidence of direct cellular toxicity, this anchoring molecule collected in the anterior vitreous, partially obscuring retinal visualization and eliciting a mild chronic microglial/macrophage inflammatory response. These studies provide a plausible approach to the development of novel non-covalent methods of binding, retention, and release of antibody-based therapeutics in the vitreous.