5′‐O‐P‐N Linkages of Phosphoramidate Nucleotides: Chemical Construction and Biological Impact

The development of chemically modified nucleotides has been a transformative area of research, driving innovations in medicinal chemistry, prebiotic chemistry, and nucleic acid‐based therapeutics. Among these modifications, 5′‐O‐P‐N phosphoramide nucleotides represent an emerging class of nucleotide analogs featuring a phosphoramidate (P‐N) linkage at the 5′‐hydroxyl position. This structural alteration significantly influences their chemical reactivity, enzymatic stability, and biological function, making them valuable tools for drug design, nucleotide‐based enzymatic studies, and synthetic biology applications. In this review, we explore the synthetic methodologies employed for constructing 5′‐O‐P‐N linkages, including phosphoramidation strategies, catalytic approaches, and emerging green chemistry techniques. Additionally, this study discusses the biological implications of these modifications, focusing on their role in nucleotide metabolism, enzyme interactions, and potential as antiviral and anticancer agents. Finally, it highlights current challenges, future directions, and the expanding scope of phosphoramide nucleotides in medicinal chemistry and biotechnology.