Success in obtaining higher-order twistarenes with precise structures is very important for fundamentally understanding the relationship between the structures and physical properties/optoelectronic applications. In this research, by using the advantages from a retro-Diels–Alder process (clean reaction) and the cross-conjugated nature of the pyrene unit, a novel dodeca-twistarene was prepared for the first time. Its structure, confirmed by single-crystal XRD analysis, indicates that it possesses a twisted angle (≈30°), and two neighboring molecules in the crystal lattice are perpendicular to each other because of the twisted character and the strong intermolecular CH–π interactions. However, its basic physicochemical properties suggest its instability in air derives from its elevated HOMO energy level, although NICS calculations confirm that the pyrene units contribution poorly to the π conjugation of the overall molecule. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim