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Abstract
Synthetic anion transporters that mediate electroneutral (H+/Cl–) transport have demonstrated anti-cancer activity due to their ability to disrupt subcellular homeostatic environments. Elucidation of the cell death mechanism revealed the transporters’ ability to neutralize lysosomal pH gradients and inhibit autophagy. However, their effect on other subcellular compartments is unknown. Herein, we disclose the first subcellular targeted anionophores that accumulate in various membrane bound organelles to bias their natural propensity to depolarize lysosomes. Confocal microscopy revealed the ability of the naphthalimide-based transporters to localize within their intended membrane-bound organelles. Analogues that contained endoplasmic reticulum (ER) and lysosomal targeting motifs showed an enhanced H+/Cl– transport ability and cytotoxicity compared to non-targeted analogues. Moreover, ER and mitochondrial localization was found to enhance apoptosis in cancerous cells. Our work provides an alternative approach to the design of therapeutically focused synthetic anion transporters and an insight into possible subcellular compartment-specific effects on homeostasis.
Supplementary materials
Title
Electronic Supplementary Information
Description
Table of Contents
General methods and materials ........................................................................................................... S1
Synthesis ............................................................................................................................................... S2
Lysosomal-targeted anionophores ................................................................................................... S3
Endoplasmic reticulum-targeted anionophores ............................................................................. S11
Mitochondrial-targeted anionophores ........................................................................................... S18
Non-targeted anionophores and control ........................................................................................ S24
Characterisation data .......................................................................................................................... S31
X-Ray crystallography .......................................................................................................................... S53
1H NMR titration experiments ............................................................................................................ S55
Sample preparation ........................................................................................................................ S55
Titration procedure ......................................................................................................................... S55
Titration data fitting ........................................................................................................................ S55
Transport studies ................................................................................................................................ S68
General remarks and vesicle preparation ....................................................................................... S68
ISE-based studies ............................................................................................................................ S68
Fluorescence HPTS-based studies ................................................................................................... S74
NMDG-Cl assay ................................................................................................................................ S74
Photophysical studies ......................................................................................................................... S79
Cell culture .......................................................................................................................................... S81
Confocal microscopy ........................................................................................................................... S81
Organelle imaging ........................................................................................................................... S81
Colocalisation analysis .................................................................................................................... S81
Cell viability ......................................................................................................................................... S89
alamarBlue cell viability assay ......................................................................................................... S89
Flow cytometry ................................................................................................................................... S91
References .......................................................................................................................................... S91
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