Inhalable dry powders of microRNA-laden extracellular vesicles prepared by thin-film freeze-drying

Khaled AboulFotouh; Gonzalo Almanza; Yu-Sheng Yu; Robert Joyce; Gregory J Davenport; Chris Cano; Robert O Williams Iii; Maurizio Zanetti; Zhengrong Cui

doi:10.1016/j.ijpharm.2023.123757

Inhalable dry powders of microRNA-laden extracellular vesicles prepared by thin-film freeze-drying

Int J Pharm. 2024 Feb 15:651:123757. doi: 10.1016/j.ijpharm.2023.123757. Epub 2023 Dec 30.

Authors

Khaled AboulFotouh¹, Gonzalo Almanza², Yu-Sheng Yu³, Robert Joyce⁴, Gregory J Davenport⁵, Chris Cano⁵, Robert O Williams Iii⁶, Maurizio Zanetti⁷, Zhengrong Cui⁸

Affiliations

¹ Division of Molecular Pharmaceutics and Drug Delivery, College of Pharmacy, The University of Texas at Austin, Austin, TX 78712, USA; Department of Pharmaceutics, Faculty of Pharmacy, Assiut University, Assiut 71526, Egypt.
² The Laboratory of Immunology, Department of Medicine and Moores Cancer Center, University of California San Diego, La Jolla, CA, 92093, USA; FutuRNA Pharmaceuticals, Inc., La Jolla, CA 92037, USA.
³ Division of Molecular Pharmaceutics and Drug Delivery, College of Pharmacy, The University of Texas at Austin, Austin, TX 78712, USA; Department of Chemical Engineering, National Taiwan University, Taipei 10617, Taiwan.
⁴ FutuRNA Pharmaceuticals, Inc., La Jolla, CA 92037, USA.
⁵ TFF Pharmaceuticals, Inc., 1751 River Run, Suite 400, Fort Worth, TX 76107, USA.
⁶ Division of Molecular Pharmaceutics and Drug Delivery, College of Pharmacy, The University of Texas at Austin, Austin, TX 78712, USA.
⁷ The Laboratory of Immunology, Department of Medicine and Moores Cancer Center, University of California San Diego, La Jolla, CA, 92093, USA; FutuRNA Pharmaceuticals, Inc., La Jolla, CA 92037, USA. Electronic address: mzanetti@health.ucsd.edu.
⁸ Division of Molecular Pharmaceutics and Drug Delivery, College of Pharmacy, The University of Texas at Austin, Austin, TX 78712, USA. Electronic address: Zhengrong.cui@austin.utexas.edu.

PMID: 38160992
DOI: 10.1016/j.ijpharm.2023.123757

Abstract

Extracellular vesicles (EVs) are endogenous vesicles that comprise a variety of submicron vesicular structures. Among these, exosomes have been widely investigated as delivery systems for small and large molecules. Herein, the thin-film freeze-drying technology was utilized to engineer aerosolizable dry powders of miR-335-laden induced EVs (iEV-335) generated in B cells for potential delivery into the lung to treat primary lung cancer and/or pulmonary metastases. The size distribution, structure, and morphology of iEV-335 were preserved after they were subjected to thin-film freeze-drying with the proper excipients. Importantly, iEV-335, in liquid or reconstituted from thin-film freeze-dried powders, were equally effective in downregulating SOX4 gene expression in LM2 human triple-negative mammary cancer cells. The iEV-335 dry powder compositions showed mass median aerodynamic diameters (MMAD) of around 1.2 µm with > 60 % of the emitted doses had an MMAD of ≤ 3 µm, indicating that the powders can potentially achieve efficient deposition within the alveolar region following oral inhalation, which is desirable for treatment of primary lung cancer and pulmonary metastases. Overall, it is concluded that it is feasible to apply thin-film freeze-drying to prepare aerosolizable dry powders of iEVs for pulmonary delivery.

Keywords: Cancer; Extracellular vesicles; Freeze drying; MicroRNA; Powders; Pulmonary delivery; SOX4.

MeSH terms

Administration, Inhalation
Dry Powder Inhalers
Extracellular Vesicles*
Freeze Drying
Humans
Lung Neoplasms*
MicroRNAs*
Particle Size
Powders / chemistry
Respiratory Aerosols and Droplets
SOXC Transcription Factors

Substances

Powders
MicroRNAs
SOX4 protein, human
SOXC Transcription Factors