New lupeol long-chain alkanoic ester 1 and lupeol β-hydroxy fatty acid esters 2c,d (laevigatins I and II) in a mixture with the previously isolated procrims a and b (2a,b) were isolated together with lupeol 3 and lupeol acetate 4 from the latex of Periploca laevigata collected in Tunisia. Their structures were elucidated by extensive spectroscopic methods including 1D (1H, 13C and DEPT 135), 2D-NMR experiments, (1H–1H COSY and NOESY), EI–MS, MALDI-TOF and GC analysis. Anti-acetylcholinesterase activity of most isolated compounds was evaluated and showed that lupeol (3) was the best inhibitor of AChE.