This chapter presents a comprehensive study of Silene popovii Schischk., an endemic species of Uzbekistan, focusing on its phytochemical composition, particularly phytoecdysteroids, and their potential pharmacological applications. The work begins with an overview of the species’ botanical characteristics and ethnopharmacological background. Detailed extraction and chromatographic techniques led to the isolation of ten phytoecdysteroids and a newly identified derivative, 20-hydroxyecdysone-25-O-benzoate. Structural elucidation was achieved through NMR spectroscopy and chromatographic profiling. The chapter explores the biosynthetic pathways, structural diversity, and ecological functions of phytoecdysteroids, alongside their pharmacological effects such as anabolic, neuroprotective, antidiabetic, antioxidant, and immunomodulatory activities. Attention is given to SAR and QSAR analyses, which inform these compounds’ structure-activity relationship and drug-likeness, supported by molecular docking and ADMET predictions. The discovery of potential triterpenoids in S. popovii further expands its metabolic profile, highlighting a synergy of secondary metabolites with therapeutic value. Additionally, their agricultural relevance is addressed through their roles in promoting plant growth and offering eco-friendly pest resistance. This chapter positions S. popovii as a valuable natural source of bioactive compounds and a promising candidate for future developments in pharmacology, nutraceuticals, and sustainable agriculture.