Abstract
Objective and Impact Statement. There is a need to develop platforms delineating inflammatory biology of the distal human lung. We describe a platform technology approach to detect in situ enzyme activity and observe drug inhibition in the distal human lung using a combination of matrix metalloproteinase (MMP) optical reporters, fibered confocal fluorescence microscopy (FCFM), and a bespoke delivery device. Introduction. The development of new therapeutic agents is hindered by the lack of in vivo in situ experimental methodologies that can rapidly evaluate the biological activity or drug-target engagement in patients. Methods. We optimised a novel highly quenched optical molecular reporter of enzyme activity (FIB One) and developed a translational pathway for in-human assessment. Results. We demonstrate the specificity for matrix metalloproteases (MMPs) 2, 9, and 13 and probe dequenching within physiological levels of MMPs and feasibility of imaging within whole lung models in preclinical settings. Subsequently, in a first-in-human exploratory experimental medicine study of patients with fibroproliferative lung disease, we demonstrate, through FCFM, the MMP activity in the alveolar space measured through FIB One fluorescence increase (with pharmacological inhibition). Conclusion. This translational in situ approach enables a new methodology to demonstrate active drug target effects of the distal lung and consequently may inform therapeutic drug development pathways.
Highlights
Respiratory diseases are one of the leading causes of morbidity and mortality worldwide [1]
To determine the clinical tractability of this approach and for the compound to be cleaved at physiologically relevant/disease levels of matrix metalloproteinase (MMP), we evaluated the propensity for FIB One to be cleaved by MMPs 2, 9, and 13 in primary human lung tissue and by stimulated human neutrophils
We report the first in-human pulmonary optical molecular imaging of enzyme activity, through fluorescence increase of an MMP reporter, and dynamic elucidation of drugtarget engagement in the distal alveolar regions of the human lung
Summary
Respiratory diseases are one of the leading causes of morbidity and mortality worldwide [1]. The recent COVID-19 global pandemic has highlighted the need for rapid approaches to assess drugs that cause respiratory morbidity and mortality [3]. The evaluation of early-phase clinical trials in human pulmonary disease has relied upon measurements, which include pulmonary function measurements, sixminute walk tests, and imaging such as computerized tomography (CT) [4]. These investigations can inform on functional and structural abnormalities but are delayed surrogates of active disease and offer little insight into. We developed a novel optical imaging approach using a combination of an existing miniaturized fibred confocal fluorescent microscopy (FCFM) system, a bronchoscope compatible delivery device, a new molecular probe, and a drug inhibitor to demonstrate real-time drug-target engagement in the distal lung of patients
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