Stereoselective Degradation of Diacylglycerol Kinases Potentiate T cell Activation and Tumor Cell Cytotoxicity

  • bioRxiv. 2025 Dec 12:2025.12.09.692983. doi: 10.64898/2025.12.09.692983.
Minhaj Shaikh  1 ,  Surya Pravo Mookherjee  1 ,  Claire C Weckerly  2 ,  Adam H Libby  3  4 ,  Aizhen Xiao  5 ,  Yunge Zhao  5 ,  Sagar D Vaidya  1 ,  AeRyon Kim  6 ,  Zhihong Li  1 ,  Madeleine L Ware  1 ,  Michelle Marants  1 ,  Olivia L Murtagh  1 ,  Wesley J Wolfe  1 ,  Timothy Nj Bullock  6 ,  Benjamin W Purow  5 ,  Gerald R V Hammond  2 ,  Ku-Lung Hsu  1
Affiliations
  • 1. Department of Chemistry, University of Texas at Austin, Austin, TX, 78712, USA.
  • 2. Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
  • 3. Department of Chemistry, University of Virginia, Charlottesville, VA 22904, USA.
  • 4. Drug Discovery Center, UVA Comprehensive Cancer Center, University of Virginia, Charlottesville, VA, 22904, USA.
  • 5. Department of Neurology, University of Virginia, Charlottesville, VA 22908, USA.
  • 6. Department of Pathology, University of Virginia, Charlottesville, VA 22908, USA.
PMID: 41427359 DOI: 10.64898/2025.12.09.692983
Abstract

Stereoselective recognition is a powerful means to differentiate selective versus non-specific activity of small molecules in complex biological systems. Here, we disclose stereochemically defined, sulfonyltriazole inhibitors of the lipid enzyme diacylglycerol kinase-alpha (DGKα), a key metabolic checkpoint for T cell effector function. Acute treatment with the covalent DGKα inhibitor AHL-7160 recruited endogenous DGKα to the plasma membrane in a stereoselective and isozyme-specific manner. The membrane translocation activity of AHL-7160 correlated with blockade of cellular phosphatidic acid production and potentiation of primary T cell-mediated killing of a glioblastoma cell line. Quantitative chemoproteomics revealed Y669 and K411 as sites of AHL-7160 modification on endogenous DGKα in cells. Extended treatments resulted in proteasome-dependent and proteome-wide selective degradation of DGKα in T cells. Collectively, these findings establish covalent DGKα ligands as potent Molecular Glues with translational potential in immunotherapy.

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