2. Academic Validation
  3. Discovery of 1-(4-cyanopyrimidin-2-yl)-1H-pyrazole-4-carboxylic acids as potent xanthine oxidase inhibitors via molecular cleavage and reassembly of allopurinol as a key strategy

Discovery of 1-(4-cyanopyrimidin-2-yl)-1H-pyrazole-4-carboxylic acids as potent xanthine oxidase inhibitors via molecular cleavage and reassembly of allopurinol as a key strategy

  • Bioorg Chem. 2026 Mar:170:109481. doi: 10.1016/j.bioorg.2026.109481.
Han Wang 1,  Jinxin Li 2,  Tong Liu 3,  Qingbo Song 2,  Qi Li 4,  Qing Mao 2,  Xuefeng Fu 2,  Xiwen Dai 2,  Jiahui Wang 2,  Xiang Shi 5,  Bing Zhang 6,  Yue Zhu 7,  Tingjian Zhang 8
Affiliations

Affiliations

  • 1 Department of Orthopedics, The First Affiliated Hospital, China Medical University, Shenyang 110000, China; School of Pharmacy, China Medical University, Shenyang 110122, China.
  • 2 Key Laboratory of Structure-Based Drugs Design & Discovery of Ministry of Education, School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016, China.
  • 3 School of Pharmacy, China Medical University, Shenyang 110122, China.
  • 4 Department of Pharmacology, Shenyang Pharmaceutical University, Shenyang 110016, China.
  • 5 School of Pharmacy, Anhui Medical University, Hefei 230032, China. Electronic address: [email protected].
  • 6 Key Laboratory of Structure-Based Drugs Design & Discovery of Ministry of Education, School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016, China. Electronic address: [email protected].
  • 7 Department of Orthopedics, The First Affiliated Hospital, China Medical University, Shenyang 110000, China. Electronic address: [email protected].
  • 8 School of Pharmacy, China Medical University, Shenyang 110122, China. Electronic address: [email protected].
PMID: 41520617 DOI: 10.1016/j.bioorg.2026.109481
Abstract

Xanthine Oxidase (XO) is a critical target for the treatment of hyperuricemia and gout. In this work, we employed a structural exploration of allopurinol via a molecular cleavage and reassembly strategy, leading to the development of 1-(4-cyanopyrimidin-2-yl)-1H-pyrazole-4-carboxylic acid derivatives as novel XO inhibitors. Structure-activity relationship analysis demonstrated that the coexistence of a polar group (e.g., OH or NH) at the 6-position of the pyrimidine and an amino group at the 5-position of the pyrazole plays a critical role for the XO inhibitory potency. Notably, the optimized compound B33 exhibited the highest XO inhibitory activity, with an IC50 value of 0.0034 μM, indicating an approximately two-fold improvement compared to febuxostat (IC50 = 0.0066 μM). Enzyme kinetic assay suggested that compound B33 exhibited a mixed-type inhibition mode toward XO. Molecular docking and molecular dynamics simulations reveal that the Ser876 forms a hydrogen bond with the NH group of B33, serving as a novel anchor residue. Furthermore, in vivo hypouricemic studies demonstrated that compound B33 significantly reduced serum uric acid level by 51.9% in mice following an oral dose of 5 mg/kg. Finally, in silico methods were employed to evaluate the drug-like properties of compound B33. Collectively, these findings demonstrate the value of the 1-(4-cyanopyrimidin-2-yl)-1H-pyrazole-4-carboxylic acid scaffold in designing non-purine XO inhibitors and identify compound B33 as a promising lead for developing anti-hyperuricemia agents.

Keywords

Molecular cleavage and reassembly; Pyrazole; Pyrimidine; Rational design; Xanthine oxidase.

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