Work

Development of a high-throughput assay format to identify inhibitors of HAX1–CLPB interaction

Year: 2026

Type: article

Abstract: The HAX1–CLPB protein–protein interaction (PPI) is critical for maintaining mitochondrial proteostasis. Since perturbation of this important axis is associated with premature death in myeloid progenitor cells and severe congenital neutropenia, we hypothesized that targeting HAX1-CLPB may have therapeutic value in myeloid malignancies. Here, we established a fluorescence polarization (FP)-based high-throughput screening (HTS) platform to identify HAX1–CLPB inhibitors. Primary screening of 40,640 compounds yielded 112 initial hits, one of them being Z16519514 ( N -methyl-2-(7-thia-9,11-diazatricyclo[6.4.0.02,6]dodeca-1(12), 2(6),8,10-tetraen-12-yloxy)acetamide).Orthogonal validation using a microscale thermophoresis (MST) competition assay confirmed this PPI inhibitor while ligand-observed Nuclear Magnetic Resonance (NMR) spectroscopy and surface plasmon resonance (SPR) confirmed target engagement on CLPB. This work provides the first biochemically validated small-molecule inhibitor of the HAX1–CLPB interaction, establishing a foundation for developing more potent and cell-active modulators targeting mitochondrial proteostasis and cell death in cancer.

Source: Results in Chemistry

DOI: 10.1016/j.rechem.2026.103765