Effective annotation of in vivo drug metabolites using liquid chromatography-mass spectrometry (LC-MS) remains a formidable challenge. Herein, a metabolic reaction-based molecular networking (MRMN) strategy is introduced, which enables the "one-pot" discovery of prototype drugs and their metabolites. MRMN constructs networks by matching metabolic reactions and evaluating MS² spectral similarity, incorporating innovations and improvements in feature degradation of MS² spectra, exclusion of endogenous interference, and recognition of redundant nodes. A minimum 75% correlation between structural similarity and MS² similarity of neighboring metabolites was ensured, mitigating false negatives due to spectral feature degradation. At least 79% of nodes, 49% of edges, and 97% of subnetworks were reduced by an exclusion strategy of endogenous ions compared to the Global Natural Products Social Molecular Networking (GNPS) platform. Furthermore, an approach of redundant ions identification was refined, achieving a 10%-40% recognition rate across different samples. The effectiveness of MRMN was validated through a single compound, plant extract, and mixtures of multiple plant extracts. Notably, MRMN is freely accessible online at https://yaolab.network, broadening its applications.

With the increasing difficulty of traditional chemical drug research and development,peptide drugs have gradually become a hot spot in drug research and development due to their advantages of high specificity,significant efficacy,easy metabolism and low toxicity.This review systematically expounds the physicochemical properties,main advantages and limitations of peptide drugs,and summarizes the currently common strategies for structural modification and delivery.It focuses on the application and target of approved peptide drugs in various diseases such as diabetes,cancer,bacterial and viral infections,multiple sclerosis,and osteoporosis.Furthermore,the research analyzes the challenges in the research and development of peptide drugs,including poor in vivo stability,low bioavailability,and limited routes of administration.It also discusses the prospects of new technologies based on molecular modification,nanodelivery systems,and computer-aided design.In summary,peptide drugs have shown unique advantages in multi-field therapy,but they still need to break through bottlenecks in preparation,delivery and drug resistance to provide new ideas and directions for future precision therapy.