Structure-Based Development of a Covalent Inhibitor Targeting Streptococcus Pyogenes over Staphylococcus Aureus Sortase A

  • Chemistry. 2025 Mar 12:e202500464. doi: 10.1002/chem.202500464.
Hailing Zhou  1  2 Ziqi Yuan  1  2 Xiang-Na Guan  1  2 Chuan Yue  3 Wei Wu  1  2 Lefu Lan  4  2 Jianhua Gan  5 Tao Zhang  1  2 Cai-Guang Yang  1  3  4  2  6
Affiliations
  • 1. State Key Laboratory of Drug Research, Centre for Chemical Biology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China.
  • 2. University of Chinese Academy of Sciences, Beijing, 100049, China.
  • 3. School of Chinese Materia Medica, Nanjing University of Chinese Medicine, Nanjing, 210023, China.
  • 4. School of Pharmaceutical Science and Technology, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou, 310024, China.
  • 5. School of Life Sciences, Fudan University, Shanghai, 200433, China.
  • 6. Shandong Laboratory of Yantai Drug Discovery, Bohai Rim Advanced Research Institute for Drug Discovery, Yantai, 264117, China.
Abstract

Sortase A (SrtA), a cysteine transpeptidase critical for surface protein anchoring in Gram-positive pathogens, represents an attractive antivirulence target. While covalent SrtA inhibitors show therapeutic potential, existing compounds lack species selectivity. Through structure-guided design, we developed T10, a covalent inhibitor selectively targeting Streptococcus pyogenes SrtA (SpSrtA) over Staphylococcus aureus SrtA (SaSrtA). Molecular docking revealed that shortening a "C=C" bond in lead compound ML346 eliminated SaSrtA inhibition due to steric hindrance from W194, while maintaining SpSrtA binding. X-ray crystallography confirmed T10's covalent modification of Cys208 in SpSrtA. T10 demonstrated two fold enhanced inhibitory potency and species-specific disruption of M-protein anchoring and biofilm formation in Streptococcus pyogenes, without affecting Staphylococcus aureus viability. In a Galleria mellonella Infection model, T10 conferred potent protection against lethal Infection. This work demonstrates the development of narrow-spectrum antivirulence agents through a structure-based rational strategy.

Keywords
Sortase A; Streptococcus pyogenes; covalent inhibitors; narrow-spectrum anti-virulence agent; structure-activity relationships.
Products