A multicomponent reaction-enabled discovery platform reveals a TXNRD1-targeted ferroptosis inducer
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PUBLIC SUMMARY
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A multicomponent reaction-based platform enabled efficient discovery of covalent inhibitors.
HF-4aa was identified as a covalent probe targeting antioxidant enzyme TXNRD1 with nanomolar potency.
HF-4aa inhibits TXNRD1, triggers redox imbalance and ferroptosis, with preclinical antitumor activity.
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ABSTRACT
Covalent inhibitors offer durable target engagement, but their discovery is often serendipitous and constrained by limited structural diversity. We introduce a multicomponent reaction (MCR)–based platform that combines modular synthesis, phenotypic screening, and proteome-level target deconvolution to identify HF-4aa, a covalent inhibitor of thioredoxin reductase 1 (TXNRD1). Stereodivergent synthesis enabled access to all four HF-4aa stereoisomers using Rh(II)/chiral phosphoric acid cocatalysis. HF-4aa displays nanomolar potency in cancer cells and modifies TXNRD1 via selenocysteine/cysteine targeting, supported by chemoproteomics and intact-protein mass spectrometry. TXNRD1 inhibition is functionally linked to redox collapse characterized by ROS accumulation, Fe2+ elevation, and lipid peroxidation, thereby inducing ferroptotic cell death. In vivo, HF-4aa suppresses tumor growth, and its efficacy is reversed by Ferrostatin-1, confirming ferroptosis dependence. Liposomal encapsulation overcomes the short metabolic half-life of free HF-4aa, and the resulting formulation demonstrates potent antitumor activity including in paclitaxel-resistant tumor models. HF-4aa serves as a proof-of-concept covalent chemical probe for TXNRD1-directed ferroptosis induction, and this study establishes a scalable MCR-based framework for systematic covalent inhibitor discovery. -
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REFERENCES
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Fu X., Wu Q., Wu S., et al. (2026). A multicomponent reaction-enabled discovery platform reveals a TXNRD1-targeted ferroptosis inducer. The Innovation Drug Discovery 1:100011. https://doi.org/10.59717/j.xinn-drugdisc.2026.100011
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Fu X., Wu Q., Wu S., et al. (2026). A multicomponent reaction-enabled discovery platform reveals a TXNRD1-targeted ferroptosis inducer. The Innovation Drug Discovery 1:100011. https://doi.org/10.59717/j.xinn-drugdisc.2026.100011
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Figure 1.
MCR-driven Compound Synthesis and Hit Compound Discovery
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Figure 1.
Discovery of the anti-TNBC hit HF-4aa and possible mechanisms
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Figure 2.
Synthesis of Probes 6a and 6b
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Figure 2.
HF-4aa covalently targets TXNRD1 via propiolamide warhead engagement
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Figure 3.
HF-4aa promotes ferroptosis hallmarks in MDA-MB-231 cells
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Figure 4.
HF-4aa induces lipid peroxidation and ferroptotic phenotypes
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Figure 5.
HF-4aa induces ferroptosis and suppresses breast cancer growth in vivo
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Figure 6.
LLC Paclitaxel-Resistant Mouse Model. A-C Data are represented as mean ± SEM
