Acenaphtho[1,2‐a]acenaphthylene (1) was sulfurated with elemental sulfur to give a pentathiepane derivative (2). A dynamic NMR spectrum analysis revealed that the two naphthalene rings of 2 are chemically nonequivalent up to 100 °C. The pentathiepane ring of 2 was shown to adopt a chair conformation both in solution and crystals. In accordance with the NMR spectrum analysis, the sulfuration of 5‐phenylacenaphtho[1,2‐a]acenaphthylene (12) gave a pair of conformers (16a and 16b), which were isolated in pure form. The Friedel–Crafts acetylation of 2, followed by dithioacetalization with 1,2‐ethanedithiol, also gave a pair of isolable conformers (18a and 18b). These two pairs of conformers isomerized, as a result of the ring inversion, to each other in solution at room temperature in the first‐order kinetics. The activation parameters for this process, Ea, ΔH#, and ΔS#, were determined. Based on the experimental observations, a probable mechanism for the inversion process is proposed.