We herein describe the identification of four new depsides present in methanol extract of Origanum dictamnus. O. dictamnus’ (dittany) aerial parts methanol extract was subjected to semi-preparative RP-HPLC fractionation followed by identification of individual compounds in each fraction using 1D/2D NMR, MS approaches and DFT calculations. The structural data revealed that 4 of the compounds were novel hitherto unidentified molecules, whereas the other 6 corresponded to known structures belonging to the groups of monoterpenes, phenolic acids and depsides. We have additionally estimated the antioxidant capacity of the methanol extract and individual fractions using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) in vitro assay. Methanol extract exhibited significant scavenging activity, which was attributed predominantly to the depside group of phytochemicals. The scavenging activity of the new compounds is reported herein for first time. Moreover, the antimitotic activities of the methanol extract as well as that of rosmarinic acid were examined and appeared to be several fold weaker comparing to their scavenging capacity.