Optimizing Ultrasonic-Assisted Extraction Process of <i>Paralepista flaccida</i>: A Comparative Study of Antioxidant, Anticholinesterase, and Antiproliferative Activities via Response Surface Methodology and Artificial Neural Network Modeling

In this study, extraction conditions were optimized to maximize the biological activities of extracts obtained from <i>Paralepista flaccida</i>, an edible mushroom species. Extraction processes were carried out using an ultrasonically assisted system, and two different optimization approaches were used as follows: Response Surface Methodology (RSM) and Artificial Neural Network–Genetic Algorithm (ANN-GA). The antioxidant potentials of the optimized extracts were evaluated using DPPH, FRAP, TAS, TOS, and OSI parameters; anticholinesterase activities were measured against AChE and BChE enzymes; and antiproliferative activities were investigated in A549, MCF-7, and DU-145 human cancer cell lines. In addition, phenolic contents were determined by LC-MS/MS analysis. The findings revealed that the extracts obtained by the RSM method exhibited a superior biological profile compared to ANN-GA extracts in terms of antioxidant, anticholinesterase, and antiproliferative activities. The high cytotoxicity observed, particularly in the MCF-7 line, supports the anticancer potential of this extract. These results demonstrate that optimization strategies are crucial for increasing not only extract yield but also biological functionality.