Effects of Benzofuran–Chalcone, Benzofuran–Pyrazoline, and Bis-Pyrazoline Derivatives on Redox-Related Biomarkers and Fatty Acid, Vitamin, and Sterol Profiles in Saccharomyces cerevisiae

Benzofuran-based hybrid compounds have reported radical-scavenging and biological activities, but their cellular biochemical response profiles require further characterization. This study evaluated previously synthesized benzofuran–chalcone, benzofuran–pyrazoline, and bis-pyrazoline derivatives by combining cell-free assays with redox-related and lipid-associated biochemical measurements in Saccharomyces cerevisiae. Chemical radical-scavenging activity was assessed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, and in vitro lipid peroxidation-related responses were evaluated using the thiobarbituric acid-reactive substances (TBARS) assay. Cellular measurements included reduced glutathione (GSH), malondialdehyde (MDA), total protein, fatty acids, and lipophilic vitamins and sterols. Cellular experiments comprised four independent experimental sets, with statistical comparisons restricted to each treatment and its corresponding concurrently prepared control. Compounds 1d and 2b showed comparatively high DPPH radical-scavenging responses under the tested conditions, whereas the in vitro TBARS responses varied with compound and treatment level. Cellular GSH and MDA changes were not consistently parallel: 2e increased both parameters, 2f markedly decreased GSH without significantly altering MDA, and 1b and 2b decreased GSH while increasing MDA. Treatments also altered fatty acid, tocopherol, and sterol profiles relative to their respective controls. These findings show that chemical radical-scavenging responses did not consistently correspond to cellular redox-related responses and that lipid-associated measurements provided complementary biochemical information. The study therefore characterizes distinct treatment-associated response profiles rather than a uniform antioxidant or pro-oxidant effect. Unequal molar exposures and the absence of concurrent viability and mechanistic measurements preclude definitive potency rankings, structure–activity relationships, or mechanistic conclusions.