A comprehensive screening platform for aerosolizable protein formulations for intranasal and pulmonary drug delivery

Abstract

Aerosolized administration of biopharmaceuticals to the airways is a promising route for nasal and pulmonary drug delivery, but - in contrast to small molecules - little is known about the effects of aerosolization on safety and efficacy of biopharmaceuticals. Proteins are sensitive against aerosolization-associated shear stress. Tailored formulations can shield proteins and enhance permeation, but formulation development requires extensive screening approaches. Thus, the aim of this study was to develop a cell-based in vitro technology platform that includes screening of protein quality after aerosolization and transepithelial permeation. For efficient screening, a previously published aerosolization-surrogate assay was used in a design of experiments approach to screen suitable formulations for an IgG and its antigen-binding fragment (Fab) as exemplary biopharmaceuticals. Efficient, dose-controlled aerosol-cell delivery was performed with the ALICE-CLOUD system containing RPMI 2650 epithelial cells at the air-liquid interface. We could demonstrate that our technology platform allows for rapid and efficient screening of formulations consisting of different excipients (here: arginine, cyclodextrin, polysorbate, sorbitol, and trehalose) to minimize aerosolization-induced protein aggregation and maximize permeation through an in vitro epithelial cell barrier. Formulations reduced aggregation of native Fab and IgG relative to vehicle up to 50% and enhanced transepithelial permeation rate up to 2.8-fold.

Insights on animal models to investigate inhalation therapy: Relevance for biotherapeutics

“…Airways epithelial cells cultured at the air-liquid interface reproduce a physiological respiratory epithelium and may be combined with a dose-controlled aerosol-cell exposure system (ALICE CLOUD) to evaluate transepithelial absorption of aerosolized protein therapeutic (Röhm et al, 2017). Additional applications with this delivery system may include the impact of inhaled biotherapeutics on airways epithelium integrity.…”

In situ-Like Aerosol Inhalation Exposure for Cytotoxicity Assessment Using Airway-on-Chips Platforms

“…Note that due to the imaged plane of the device (i.e., PET membrane), only deposited particles are identified (compared with those deposited on the PDMS side walls). Altogether, our in vitro exposure assays capture realistic aerosol transport determinants that still remain widely absent in more traditional PM instillations or direct spraying techniques (Blank et al, 2006;Lenz et al, 2014;Röhm et al, 2017) on plates and inserts. Furthermore, our airway-on-chips highlight for the first time that exposure assays can be configured to explore localized deposition effects, capturing for example deposition outcomes pertinent to regional lung lobes (e.g., upper versus lower lobes) that span broad orientation angles across the chest cavity (Sauret et al, 2002).…”

“…Importantly, our in vitro inhalation assays deliver aerosols simultaneously across four airway-on-chips integrated within a larger conductive airway tree model to explore in situ-like aerosol deposition outcomes under physiological respiratory airflows and for various gravitational orientations. Given that current cytotoxicity assays are still widely based on either non-physiological liquid installations of PM suspensions (Hittinger et al, 2017) or alternatively direct spraying at an ALI (Blank et al, 2006;Lenz et al, 2014;Röhm et al, 2017), our airway-on-chip exposure setup is part of steadfast efforts (Artzy-schnirman et al, 2019a) leading toward a new generation of advanced in vitro lung toolkits for human inhalation toxicity assays.…”

How to characterize a nasal product. The state of the art of in vitro and ex vivo specific methods

“…In comparison, chamber configurations can deliver a uniform density through larger membranes (diameter ≥24 mm), with a reduced reading variation (Srinivasan et al, 2015). Both excised mucosa and cell layers require an equilibrium time (∼30 min or 60 min (Röhm et al, 2017;Lungare et al, 2016;Wang et al, 2017;Li et al, 2006) before performing the permeation study, which then consists of taking aliquots from the basal compartment at regular intervals (Gonçalves et al, 2016). The deposition of a reproducible and uniform amount of drug on the tissue is thus crucial as it will directly influence the gradient concentration and could thus introduce variation in the permeability (Upadhyay et al, 2017).…”