Highly fluorinated candidates containing anticancer pharmacophores like thiosemicarbazone (<b>5a</b>-<b>e</b>) and its cyclic analogues hydrazineylidenethiazolidine (<b>6a</b>-<b>e</b>), 2-aminothiadiazole (<b>7a</b>-<b>e</b>), and 2-hydrazineylidenethiazolidin-4-one (<b>8a</b>-<b>e</b>) were synthesized, and their cytotoxic activity was assayed against 60 tumor cell lines. Compounds <b>6c</b>, <b>7b</b>, and <b>8b</b> displayed the most potent activity with lower toxic effects on MCF-10a. <i>In vitro</i> phosphatidylinositol 3-kinase (PI3K) enzyme inhibition was performed. Compound <b>6c</b> displayed half-maximal inhibitory concentration (IC<sub>50</sub>, μM) values of 5.8, 2.3, and 7.9; compound <b>7b</b> displayed IC<sub>50</sub> values of 19.4, 30.7, and 73.7; and compound <b>8b</b> displayed IC<sub>50</sub> values of 77.5, 53.5, and 121.3 for PI3Kα, β, and δ, respectively. Moreover, cell cycle progression caused cell cycle arrest at the S phase for compounds <b>6c</b> and <b>8b</b> and at G1/S for compound <b>7b</b>, while apoptosis was induced. <i>In silico</i> studies; molecular docking; physicochemical parameters; and absorption, distribution, metabolism, excretion, and toxicity (ADMET) analysis were performed. The results showed that compound <b>6c</b> is the most potent one with a selectivity index (SI) of 39 and is considered as a latent lead for further optimization of anticancer agents.