Adamantoid Scaffolds for Multiple Cargo Loading and Cellular Delivery as β‐Cyclodextrin Inclusion Complexes

There is huge demand for developing guests that bind β-CD and can conjugate multiple cargos for cellular delivery. We synthesized trioxaadamantane derivatives, which can conjugate up to three cargos per guest. 1 H NMR titration and isothermal titration calorimetry revealed these guests form 1 : 1 inclusion complexes with β-CD with association constants in the order of 103 M-1 . Co-crystallization of β-CD with guests yielded crystals of their 1 : 1 inclusion complexes as determined by single-crystal X-ray diffraction. In all cases, trioxaadamantane core is buried within the hydrophobic cavity of β-CD and three hydroxyl groups are exposed outside. We established biocompatibility using representative candidate G4 and its inclusion complex with β-CD (β-CD⊂G4), by MTT assay using HeLa cells. We incubated HeLa cells with rhodamine-conjugated G4 and established cellular cargo delivery using confocal laser scanning microscopy (CLSM) and fluorescence-activated cell sorting (FACS) analysis. For functional assay, we incubated HeLa cells with β-CD-inclusion complexes of G4-derived prodrugs G6 and G7, containing one and three units of the antitumor drug (S)-(+)-camptothecin, respectively. Cells incubated with β-CD⊂G7 displayed the highest internalization and uniform distribution of camptothecin. β-CD⊂G7 showed higher cytotoxicity than G7, camptothecin, G6 and β-CD⊂G6, affirming the efficiency of adamantoid derivatives in high-density loading and cargo delivery.