Valerianofal A (2), a rare sesquiterpenoid-alkaloid hydrid, as well as two biosynthetic related compounds (1 and 3) was isolated from the roots of Valeriana officinalis var. latifolia Miq. Compound 2 represented the first germacrane-type sesquiterpenoid and tecostidine hydrid. Their structures as well as absolute configurations were elucidated by spectroscopic analysis, an alkaline hydrolysis method, and X-ray diffraction. The structure of volvalerenic acid A was revised by single-crystal X-ray diffraction. Compounds 2 and 3 displayed selective BuChE inhibitory activities with IC50 values ranging from 0.4 μg/ml to 2.8 μg/ml. The possible recognition mechanism between compound 2 and BuChE was predicted by molecular docking analysis. © 2022 Elsevier Ltd