This Research Topic was intended to focus on new insights, novel developments, current challenges, latest discoveries, recent advances, and future perspectives in Experimental Pharmacology and Drug Discovery. When launching this section, 12 years ago, we stated that it would deal with the analysis of those biological mechanisms targetable by drugs and/or affecting their action (Salomone, 2010). This analysis may be carried out at molecular, cellular, organ/organism level, with a further fourth level of analysis, individualized pharmacology, which takes into account the impact of genetics and epigenetics on drug efficacy and toxicity, to optimize drug therapy for each individual. Drug discovery may move from new chemical entities, conceived to target specific molecular and cellular processes relevant for a disease mechanism, but often takes the reverse way, which is moving from approved drugs to the study of their mechanisms, potentially exploitable to cure other, often unrelated, diseases (drug repurposing). Unexpectedly, our section has also seen a continuously growing number of submissions focusing on natural compounds, which are part of the traditional pharmacopeia in some cultures, particularly the Chinese one, but whose specific mechanisms are not known. These studies, when carried out according to the standards of modern pharmacology (pure substances, precisely known concentrations/doses, concentration-response relationship), may provide useful information not only to understand and implement the correct use of traditional medicine but, more importantly, to provide new lead compounds, specifically directed toward relevant pathophysiological targets.