Microliter-scale in vitro aerosol-cell delivery of inhalable antisense oligonucleotide (ASO) induces "burst-like" pharmacokinetics.
6 affiliations
- 1Institute of Lung Health and Immunity, Comprehensive Pneumology Center (CPC-M) with the CPC-M bioArchive, Ludwig-Maximilians-Universität München and Helmholtz Munich, Member of the German Center for Lung Research (DZL), Germany.
- 2Centre d'Etudes des Pathologies Respiratoires / Inserm UMR1100, Tours, France.
- 3Université de Tours, Tours, France.
- 4Secarna Pharmaceuticals GmbH & Co. KG, Planegg, Germany.
- 5Université catholique de Louvain, Louvain Drug Research Institute, Advanced Drug Delivery & Biomaterials, Belgium.
- 6CHRU de Tours, Service de Pneumologie et Explorations Fonctionnelles Respiratoires, Tours, France.
Abstract
Unlike traditional submerged cell cultures, nebulized drug delivery to air-liquid interface (ALI)-cultured lung cells provides a clinically relevant route for in vitro testing of inhalable drugs. Current nebulizer systems deliver non-physiologically large drug volumes. Here, we introduce the Cloud MAX technology, characterize its accuracy of dosimetry, and apply it to toxicity, efficacy, and biokinetics or permeability testing of a locked nucleic acid-modified antisense oligonucleotide (ASO). On average the Cloud MAX delivers 44% of 3-20 μL aerosolized liquid to ALI-cultured cells, the coefficient of variation (CV) of repeated drug dosing is <11%-rel. and as little as 1 μL of drug/insert (=0.25 μL/cm2) can be delivered to ALI-cultured cells. As proof of concept for dose-controlled in vitro drug testing, we demonstrated target engagement of a LIM and Calponin Homology domain 1 (LIMCH1)-specific ASO in primary lung fibroblasts from Idiopathic Pulmonary Fibrosis (IPF) patients "shielded" by an A549 alveolar epithelial layer mimicking inhalation therapy. LIMCH1-ASO was effective in LIMCH1-silencing even at the lowest dose (0.64 nmol/cm2). Nebulization did neither impair LIMCH1-ASO efficacy nor induce cytotoxicity for the highest tested dose (3.21 nmol/cm2). The permeability (biokinetics) of nebulized ASO through the A549 cell barrier was 14-fold faster ("burst-like") than for larger volume (pipetted) ASO application. Computational modeling revealed that this well-known "burst-like" pharmaco-/biokinetics profile is due to small-volume drug application, not due to nebulizer-induced changes on barrier tightness (apparent permeability Papp ). Thus, the Cloud MAX is a substance-efficient platform for in vitro pharmacokinetics/-dynamics testing of inhalable drugs for targeted pulmonary therapy.
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BibTeX
@article{kumar2026microliterscale,
title = {{Microliter-scale in vitro aerosol-cell delivery of inhalable antisense oligonucleotide (ASO) induces "burst-like" pharmacokinetics.}},
author = {Pramod Kumar and Ashesh Chakraborty and Elias Richter and Lin Yang and Andreas Schröppel and Coralie Mergault and Larissa Rothkirch Née Knüppel and Frank Jaschinski and Marion Blayac and Rita Vanbever and Laurent Plantier and Nathalie Heuzé-Vourch and Otmar Schmid},
journal = {International journal of pharmaceutics: X},
year = {2026},
month = jul,
doi = {10.1016/j.ijpx.2026.100588},
url = {https://europepmc.org/article/MED/42999459},
note = {CC BY},
}
RIS
TY - JOUR
TI - Microliter-scale in vitro aerosol-cell delivery of inhalable antisense oligonucleotide (ASO) induces "burst-like" pharmacokinetics.
AU - Pramod Kumar
AU - Ashesh Chakraborty
AU - Elias Richter
AU - Lin Yang
AU - Andreas Schröppel
AU - Coralie Mergault
AU - Larissa Rothkirch Née Knüppel
AU - Frank Jaschinski
AU - Marion Blayac
AU - Rita Vanbever
AU - Laurent Plantier
AU - Nathalie Heuzé-Vourch
AU - Otmar Schmid
PY - 2026
DA - 2026/07/01
JO - International journal of pharmaceutics: X
DO - 10.1016/j.ijpx.2026.100588
UR - https://europepmc.org/article/MED/42999459
AB - Unlike traditional submerged cell cultures, nebulized drug delivery to air-liquid interface (ALI)-cultured lung cells provides a clinically relevant route for in vitro testing of inhalable drugs. Current nebulizer systems deliver non-physiologically large drug volumes. Here, we introduce the Cloud MAX technology, characterize its accuracy of dosimetry, and apply it to toxicity, efficacy, and biokinetics or permeability testing of a locked nucleic acid-modified antisense oligonucleotide (ASO). On average the Cloud MAX delivers 44% of 3-20 μL aerosolized liquid to ALI-cultured cells, the coefficient of variation (CV) of repeated drug dosing is <11%-rel. and as little as 1 μL of drug/insert (=0.25 μL/cm2) can be delivered to ALI-cultured cells. As proof of concept for dose-controlled in vitro drug testing, we demonstrated target engagement of a LIM and Calponin Homology domain 1 (LIMCH1)-specific ASO in primary lung fibroblasts from Idiopathic Pulmonary Fibrosis (IPF) patients "shielded" by an A549 alveolar epithelial layer mimicking inhalation therapy. LIMCH1-ASO was effective in LIMCH1-silencing even at the lowest dose (0.64 nmol/cm2). Nebulization did neither impair LIMCH1-ASO efficacy nor induce cytotoxicity for the highest tested dose (3.21 nmol/cm2). The permeability (biokinetics) of nebulized ASO through the A549 cell barrier was 14-fold faster ("burst-like") than for larger volume (pipetted) ASO application. Computational modeling revealed that this well-known "burst-like" pharmaco-/biokinetics profile is due to small-volume drug application, not due to nebulizer-induced changes on barrier tightness (apparent permeability Papp ). Thus, the Cloud MAX is a substance-efficient platform for in vitro pharmacokinetics/-dynamics testing of inhalable drugs for targeted pulmonary therapy.
SN - 2590-1567
ER -