Diastereoselective synthesis of cyclic tetrapeptide pseudoxylallemycin A illuminates the impact of base during macrolactamization

• Published in: Organic & biomolecular chemistry Vol. 21; no. 5; pp. 156 - 169
• Main Authors: Fumo, Vincent M, Roberts, R. Charlie, Zhang, Jieyu, O'Reilly, Matthew C
• Format: Journal Article
• Language: English
• Published: England Royal Society of Chemistry 01.02.2023
• Subjects: Antibacterial activity; Antibiotics; Biological Products; Chemical compounds; Chemical reactions; Chemical synthesis; Diastereoisomers; Molecular Conformation; Molecular Structure; Multidrug resistance; Natural products; NMR; Nuclear magnetic resonance; Peptides; Peptides, Cyclic - chemistry; Pharmacology; Racemization; Reagents; Selectivity; Stereoisomerism; Stereoisomers; Stereoselectivity
• ISSN: 1477-0520; 1477-0539; 1477-0539
• DOI: 10.1039/d2ob02126a

Therapeutic agents with unique molecular structures and new mechanisms of action are needed to confront the phenomenon of multidrug resistance among bacteria. Pseudoxylallemycins, cyclic tetrapeptide (CTP) natural products, have exhibited modest antibiotic activity, but their synthesis has proven challenging. Inherent ring strain in CTPs decreases the rate of cyclization in lieu of polymerization and racemization pathways, which has resulted in previous syntheses describing mixtures of diastereomers containing predominantly an undesired epimer. We have optimized the cyclization step of pseudoxylallemycin A to favor production of the natural diastereomer; notably, variation of the base, temperature, and solvent with peptide coupling reagent propylphosphonic anhydride (T3P) afforded exquisite selectivity for the natural product in as high as 97 : 3 DR, and our conditions can provide the natural product in up to 32% overall yield through 8 steps. Employing weaker bases than those typically used in peptide coupling reactions led to the greatest improvement in diastereoselectivity, and these studies demonstrated that the identity of the amine base has enormous impact on the rate of C-terminal epimerization when T3P is used, a variable usually considered of lesser consequence when combined with typical amide coupling reagents. Toward fully characterizing pseudoxylallemycin stereoisomers, variable temperature NMR was described as a tool to more clearly analyze CTPs that exhibit multiple conformational states. These synthetic and spectroscopic insights were applied toward synthesizing several natural product analogues, and their antibacterial activity was examined using microdilution assays. Amine bases go beyond their typical proton shuttle role in the macrolactamization of pseudoxylallemycin, as we diastereoselectively synthesize the natural product and analogues thereof to explore their characterization and biological activity.