2. Academic Validation
  3. Structure-guided design of potent tetrazolo[1,5-a]pyrimidine-based tubulin inhibitors with in vivo antitumor activity

Structure-guided design of potent tetrazolo[1,5-a]pyrimidine-based tubulin inhibitors with in vivo antitumor activity

  • Eur J Med Chem. 2026 Jan 15;302(Pt 1):118288. doi: 10.1016/j.ejmech.2025.118288.
Yaolin Guo 1,  Quanwei Yu 2,  Cuiyu Guo 1,  Chengyong Wu 1,  Lun Tan 1,  Ruofei Zhang 1,  Yang He 3,  Aiping Tong 4,  Yuxi Wang 5
Affiliations

Affiliations

  • 1 State Key Laboratory of Biotherapy and Cancer Center, Department of Pulmonary and Critical Care Medicine, Targeted Tracer Research and Development Laboratory, Institute of Respiratory Health, State Key Laboratory of Respiratory Health and Multimorbidity, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, China.
  • 2 Heilongjiang Institute for Drug Control, Harbin, 150088, China; Department of Respiratory and Critical Care Medicine, Molecularly Targeted Research and Development Laboratory, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, China.
  • 3 Frontiers Medical Center, Tianfu Jincheng Laboratory, Chengdu, 610212, Sichuan, China; Department of Respiratory and Critical Care Medicine, Molecularly Targeted Research and Development Laboratory, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, China.
  • 4 State Key Laboratory of Biotherapy and Cancer Center, Department of Pulmonary and Critical Care Medicine, Targeted Tracer Research and Development Laboratory, Institute of Respiratory Health, State Key Laboratory of Respiratory Health and Multimorbidity, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, China; State Key Laboratory of Biotherapy and Cancer Center, Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan Province, China.
  • 5 State Key Laboratory of Biotherapy and Cancer Center, Department of Pulmonary and Critical Care Medicine, Targeted Tracer Research and Development Laboratory, Institute of Respiratory Health, State Key Laboratory of Respiratory Health and Multimorbidity, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, China; Frontiers Medical Center, Tianfu Jincheng Laboratory, Chengdu, 610212, Sichuan, China. Electronic address: [email protected].
PMID: 41172644 DOI: 10.1016/j.ejmech.2025.118288
Abstract

Inhibitors targeting the colchicine-binding site of tubulin (CBSIs) represent a promising class of antitumor agents, offering the potential to overcome multidrug resistance. Through high-throughput virtual screening (HTVS) via molecular docking (Glide) followed by structural optimization, a novel hit compound Q1 (tetrazolo[1,5-a]pyrimidine scaffold; SKOV3 IC50 = 2.20 ± 0.05 μM) was identified from the ChemDiv library. Molecular dynamics (MD) simulations revealed that the cyclohexane moiety of Q1 exhibited high flexibility, which was identified as a key factor limiting its potency. Three stages of structural modification (Parts A-C) yielded 66 derivatives. The racemic lead compound rac-Q31 (hereinafter referred to as Q31 for brevity), which bears an optimized 3,5-dimethylphenyl group, exhibited potent antitumor activity with an IC50 of 24.6 ± 1.4 nM against SKOV3 cells. A co-crystal structure (PDB: 9LSE) confirmed that Q31 binds stably within the colchicine site, forming a hydrogen bond with α-Thr179 and key hydrophobic interactions, including those with α-Leu240 and β-Ile316. (S)-Q31 showed enhanced activity (SKOV3 IC50 = 17.2 ± 1.8 nM). In an SKOV3 xenograft model, (S)-Q31 exhibited significant antitumor efficacy (tumor growth inhibition, TGI = 74.12 % at 4 mg/kg, i.v.) with good tolerability. Mechanistically, (S)-Q31 inhibited microtubule polymerization, arrested the cell cycle at the G2/M phase, and induced Apoptosis. This study establishes (S)-Q31 as a promising CBSI lead compound and provides structural insights for novel microtubule inhibitor development.

Keywords

Colchicine-binding site inhibitor; Crystal structure; High-throughput virtual screening; Structure-based drug design; Tubulin.

Figures
Products