Preclinical drug development faces inherent limitations associated with conventional models, such as two-dimensional cell cultures and animal models, which exhibit limited physiological relevance and contribute to high attrition rates in clinical phases. NAMs, including human organoids, organ-on-chip systems, microphysiological platforms, physiologically based pharmacokinetic modeling, omics technologies, and artificial intelligence, provide an integrated and predictive framework for the assessment of organ-specific toxicities, mechanistic characterization, and human extrapolation. These tools enable dose optimization, improved risk monitoring, and reduction in animal use. Nevertheless, challenges remain regarding functional maturation, experimental reproducibility, regulatory validation, and population representativeness. The strategic and combined application of NAMs constitutes an advanced preclinical ecosystem that complements traditional approaches and holds significant potential to enhance the safety and efficacy of drug candidates in development.