Weak base drug-induced endolysosome iron dyshomeostasis controls the generation of reactive oxygen species, mitochondrial depolarization, and cytotoxicity

Peter W Halcrow; Darius N K Quansah; Nirmal Kumar; Rebecca L Solloway; Kayla M Teigen; Kasumi A Lee; Braelyn Liang; Jonathan D Geiger

doi:10.1515/nipt-2023-0021

Weak base drug-induced endolysosome iron dyshomeostasis controls the generation of reactive oxygen species, mitochondrial depolarization, and cytotoxicity

NeuroImmune Pharm Ther. 2024 Jan 11;3(1):33-46. doi: 10.1515/nipt-2023-0021. eCollection 2024 Mar.

Authors

Peter W Halcrow¹, Darius N K Quansah¹, Nirmal Kumar¹, Rebecca L Solloway¹, Kayla M Teigen¹, Kasumi A Lee¹, Braelyn Liang¹, Jonathan D Geiger¹

Affiliation

¹ Department of Biomedical Sciences, School of Medicine and Health Sciences, University of North Dakota, Grand Forks, ND, USA.

Abstract

Objectives: Approximately 75 % of marketed drugs have the physicochemical property of being weak bases. Weak-base drugs with relatively high pK_a values enter acidic organelles including endosomes and lysosomes (endolysosomes), reside in and de-acidify endolysosomes, and induce cytotoxicity. Divalent cations within endolysosomes, including iron, are released upon endolysosome de-acidification. Endolysosomes are "master regulators of iron homeostasis", and neurodegeneration is linked to ferrous iron (Fe²⁺)-induced reactive oxygen species (ROS) generation via Fenton chemistry. Because endolysosome de-acidification-induced lysosome-stress responses release endolysosome Fe²⁺, it was crucial to determine the mechanisms by which a functionally and structurally diverse group of weak base drugs including atropine, azithromycin, fluoxetine, metoprolol, and tamoxifen influence endolysosomes and cause cell death.

Methods: Using U87MG astrocytoma and SH-SY5Y neuroblastoma cells, we conducted concentration-response relationships for 5 weak-base drugs to determine EC₅₀ values. From these curves, we chose pharmacologically and therapeutically relevant concentrations to determine if weak-base drugs induced lysosome-stress responses by de-acidifying endolysosomes, releasing endolysosome Fe²⁺ in sufficient levels to increase cytosolic and mitochondria Fe²⁺ and ROS levels and cell death.

Results: Atropine (anticholinergic), azithromycin (antibiotic), fluoxetine (antidepressant), metoprolol (beta-adrenergic), and tamoxifen (anti-estrogen) at pharmacologically and therapeutically relevant concentrations (1) de-acidified endolysosomes, (2) decreased Fe²⁺ levels in endolysosomes, (3) increased Fe²⁺ and ROS levels in cytosol and mitochondria, (4) induced mitochondrial membrane potential depolarization, and (5) increased cell death; effects prevented by the endocytosed iron-chelator deferoxamine.

Conclusions: Weak-base pharmaceuticals induce lysosome-stress responses that may affect their safety profiles; a better understanding of weak-base drugs on Fe²⁺ interorganellar signaling may improve pharmacotherapeutics.

Keywords: ADMET; cell death; endolysosome ferrous iron; mitochondrial membrane potential; reactive oxygen species; weak base drugs.