NQO1-responsive Trimethyl lock benzoquinone: A cleavable linker strategy for antibody-drug conjugates

  • J Control Release. 2025 Dec 11:390:114534. doi: 10.1016/j.jconrel.2025.114534.
Zihao Wang  1 ,  Kai Liu  2 ,  Chao Shang  3 ,  Xiangyu Yan  4 ,  Yuchao Dong  1 ,  Dapeng Li  5 ,  Yiquan Li  3 ,  Cuiling Zhang  3 ,  Wei Ren  1 ,  Chengyuan Ma  5 ,  Huan Wang  6 ,  Xiao Li  7 ,  Shiyong Fan  8 ,  Wu Zhong  9
Affiliations
  • 1. National Engineering Research Center for the Emergency Drug, Beijing 100039, China.
  • 2. Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun, Jilin 130122, China; Department of Neurosurgery, The First Hospital of Jilin University, Changchun, Jilin 130000, China.
  • 3. Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun, Jilin 130122, China.
  • 4. State Key Laboratory of Powder Metallurgy, Central South University, Changsha, Hunan 410083, China.
  • 5. Department of Neurosurgery, The First Hospital of Jilin University, Changchun, Jilin 130000, China.
  • 6. State Key Laboratory of Rare Earth Resource Utilization and Laboratory of Chemical Biology, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin 130022, China. Electronic address: [email protected].
  • 7. Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun, Jilin 130122, China. Electronic address: [email protected].
  • 8. National Engineering Research Center for the Emergency Drug, Beijing 100039, China. Electronic address: [email protected].
  • 9. National Engineering Research Center for the Emergency Drug, Beijing 100039, China. Electronic address: [email protected].
PMID: 41389965 DOI: 10.1016/j.jconrel.2025.114534
Abstract

Antibody-drug conjugate (ADC) efficacy is hindered by premature systemic payload release and insufficient tumor activation. Here, an NQO1-responsive linker utilizing trimethyl lock benzoquinone is developed to create NRTL-24, a glioma-targeting ADC. In vitro studies showed NRTL-24 maintained plasma stability while achieving NQO1-dependent MMAE release. It demonstrated potent cytotoxicity in EGFR/NQO1-high glioma cells (U251 IC50 = 0.03 nM; T98G IC50 = 0.06 nM), comparable to free MMAE (U251 IC50 = 0.01 nM; T98G IC50 = 0.02 nM). Notably, toxicity dropped significantly in EGFR/NQO1-low normal cells (HMC3 IC50 = 0.58 mM), yielding >10,000 selectivity index. In vivo, NRTL-24 demonstrated superior tumor-targeting ability, significantly prolonging median survival compared to untreated and temozolomide-treated groups while maintaining favorable safety. This NQO1-responsive linker technology enables precise payload activation in tumor while minimizing off-target effects, establishing NRTL-24 as a promising therapeutic candidate. The strategy provides a blueprint for enzyme-targeted ADC development through tumor- selective linker design.

Keywords
Antibody-drug conjugates; Antitumor drug development; Cleavable linker; Drug delivery; Glioma therapy; NQO1-responsible linker; Targeted therapy; Trimethyl lock benzoquinone.
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