Identification of the target ANTXR1 and covalent acylation mechanism of 8-esterified cycloberberines against cancer

  • Eur J Med Chem. 2026 Oct 5:315:118965. doi: 10.1016/j.ejmech.2026.118965.
Jia Tang  1 ,  Xican Ma  1 ,  Xuelei Wang  1 ,  Guimin Xia  1 ,  Xintong Zhang  1 ,  Zhihui Yu  1 ,  Jingyang Zhu  1 ,  Xinyi Li  1 ,  Zhiyun Wu  1 ,  Runze Meng  1 ,  Yanan Ni  1 ,  Tianyun Fan  2 ,  Yinghong Li  3 ,  Danqing Song  4
Affiliations
  • 1. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China; State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China.
  • 2. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China; State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China. Electronic address: [email protected].
  • 3. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China; State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China. Electronic address: [email protected].
  • 4. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China; State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China. Electronic address: [email protected].
PMID: 42143941 DOI: 10.1016/j.ejmech.2026.118965
Abstract

An ester group can serve as a covalent warhead to acylate amino acid residues around the active cavity of the target protein, thereby silencing its biological function. In this study, taking 8-(benzoyloxy) cycloberberine (1) as the lead, twenty-seven 8-esterified cycloberberine (CBBR) derivatives were continuously synthesized and evaluated for their anti-tumor activities. Among them, compound 8a exhibited an appealing potency against a variety of tumor cells with IC50 values ranging 1.91-5.09 μM, significantly outperforming the lead 1. Cell cycle analysis showed 8a-induced G2/M phase arrest, which suggested ANTXR1 as a candidate target via thermal proteome profiling (TPP) assay. Further cellular thermal shift assay (CETSA) and surface plasmon resonance (SPR) analysis identified ANTXR1 as a direct target of 8a (KD = 2.88 μM). LC-MS/MS, molecule docking and molecular dynamics simulations (MD) demonstrated that the 8-adamantane acetyl in 8a could undergo an acylation reaction with Ser229 of ANTXR1, thereby inhibiting tumor cell proliferation. Therefore, compound 8a is a covalent inhibitor of ANTXR1, and introducing an ester group as the chemical warhead represents a highly effective structural modification strategy in medicinal chemistry.

Keywords
ANTXR1; Acylation covalent; Anti-tumor activity; Esterified cycloberberine; Synthesis.
Products