Integrated platform for accelerating therapeutic macrocyclic peptide purification in high-throughput medicinal...

Peptides are generating significant interest in drug discovery due to their unique therapeutic profile when compared to small molecules. As pharmaceutical companies establish more peptide-based discovery programs, innovative workflows that are both efficient and adaptable are necessary. Incorporating advanced digital tools and state-of-the-art instrumentation to accommodate high-throughput design, synthesis and purification have become increasingly valuable. Purification has historically been a bottleneck for peptide-based drug discovery programs and would benefit from a unified platform that integrates data across systems and automates processes. Here we demonstrate a streamlined platform for synthetic macrocyclic peptides from submission to final reporting using a commercial software tool that enables chromatographic method prediction based on chemical quantitative structure-retention relationships (QSRR) paired with a reversed-phase high-performance liquid chromatography (RP-HPLC) semi-preparative system. The system accommodates a wide range of crude peptide sample size from mini-scale to larger reaction scales (0.25 to 25.0 μmol) without sacrificing resolution, purity, or recovery. A triple detector configuration is utilized for obtaining a more comprehensive peptide impurity profile and direct quantitation with charged aerosol detection (CAD) is possible for low yielding samples. Example use cases with macrocyclic peptides that impact insulin sensitivity are demonstrated. The flexibility of this workflow is also applicable to many different types of modalities which require purification, from small molecules to bioconjugates. These optimized strategies result in a substantial reduction in organic solvent consumption, toxic waste production, and overall operational cost when compared with legacy methods and shorten the design-make-test (DMT) cycle of new drugs.