The conformational analysis of a series of tri- and tetrahydroxyazepanes 1–6 designed as noeuromycin mimics has been carried out using ¹H NMR spectroscopy assisted by molecular mechanics, molecular dynamics and Monte Carlo calculations. A marked flexibility for these compounds has been found. Superimposition of the branched azepanes with known glycosidase inhibitors (DMJ, DNJ, IFG and noeuromycin) was in accordance with the glycosidase inhibition profile of the polyhydroxylated azepanes. Additional STD experiments confirmed the potency and competitive character of azepane 3 towards almond β-glucosidase.