Lysosomal Targeting of β‐Cyclodextrin

β‐Cyclodextrin (β‐CD) and derivatives are approved therapeutics in >30 clinical settings. β‐CDs have also shown promise as therapeutics for treatment of some lysosomal storage disorders, such as Niemann‐Pick disease type C, and other disease states which involve metabolite accumulation in the lysosome. In these cases, β‐CD activity relies on transport to the lysosome, wherein it can bind hydrophobic substrate and effect extraction. The post‐translational attachment of N‐glycans terminated in mannose‐6‐phosphate (M6P) residues is the predominant method by which lysosomal enzymes are targeted to the lysosome. In this work we covalently attach a synthetic biantennary bis‐M6P‐terminated N‐glycan to β‐CD and study the effect of the added glycans in a mammalian cell line. The formation of a host guest complex with a Cy5 fluorophore allows study of both cellular internalisation and transport to the lysosome by fluorescence microscopy. Results indicate that the rates of both internalisation and lysosomal transport are increased by the attachment of M6P‐glycans to β‐CD, indicating that M6P‐glycan conjugation may improve the therapeutic effectiveness of β‐CD for the treatment of disorders involving hydrophobic metabolite accumulation in the lysosome. Cyclodextrins. Attachment of a bis‐mannose‐6‐phosphate terminated glycan to β‐cyclodextrin increases both cellular internalisation and transport to the lysosome, indicating potential application for improved treatments for lysosomal metabolite accumulation