The safety and efficacy of naturally occurring anticancer agents and their derivatives such as thymoquinone (TQ) and thymohydroquinone (THQ) have gained a rapidly growing interest. In an attempt to develop novel anticancer agents with superior activity, TQ was allowed to react with hydrazine hydrate, producing hydrazino thymohydroquinone 3. This new intermediate was subsequently reacted with various isocyanates, isothiocyanates, and acyl halides, affording three series of semicarbazone, semithiocarbazone, and acyl hydrazone derivatives, respectively. Subsequently, the anticancer activity of all the newly synthesised compounds against a panel of cancer cell lines was evaluated. Initial screening of the ability of the test compounds to inhibit cancer cell viability using the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay revealed that compounds 5d and 6 exerted better activity against breast cancer than TQ, with half-maximal inhibitory concentration (IC50) values of 9.6 and 10.0 μM, respectively. MTT results were confirmed by the ability of these compounds to elicit apoptotic cell death through the activation of caspase 3/7 enzymes. Together, the present work provided a novel class of THQ-based derivatives with potent anticancer and apoptosis properties, thereby warranting further optimisation of these derivatives as novel members in cancer treatment protocols.