Abstract / Summary
Abstract Background. Ferroptosis resistance in KRAS-mutant colorectal cancer (CRC) runs through at least two independent axes — the GPX4–glutathione pathway and the FSP1–CoQ10–NAD(P)H pathway — and KRAS-mutant cells specifically upregulate FSP1 via MAPK and NRF2. Selective inhibition of GPX4 or SLC7A11 alone leaves FSP1 resistance intact, and no single compound with dual GPX4 and FSP1 activity has been reported. Methods. We ran cross-target docking of eleven compounds against the crystal structure of human FSP1 (PDB 8WIK, 2.00 Å, with 6-OH-FAD and NAD) using AutoDock Vina 1.2.7 [8], targeting the CoQ10/catalytic pocket and the NAD-binding allosteric cavity. The set comprised two GPX4 candidates (JKE-1674, ClAcNH_001) from prior docking against PDB 7KTC, one SLC7A11 candidate (PipAryl_005) from PDB 7P9U, two reference FSP1 inhibitors (iFSP1, FSEN1), and six de novo CoQ10-pharmacophore scaffolds. All compounds were profiled for Lipinski compliance, PAINS status, and ADMET in RDKit. Vertical electron affinities were computed at r2SCAN-3c/CPCM(water) in ORCA 5.0 [26] to assess the redox liability of quinone scaffolds. Results. JKE-1674 (GPX4 − 8.83 kcal/mol) scored − 9.90 at the FSP1 CoQ pocket, approaching the iFSP1 reference (− 10.10). PipAryl_005 (SLC7A11 − 8.47 kcal/mol) scored − 10.60, exceeding iFSP1 at both FSP1 sites. Among the de novo scaffolds, Quinone_002 (QED 0.916, − 9.80) and CoQ_Mimic_002 (QED 0.899, − 9.20) ranked highest. Quantum-chemical calculations confirmed that naphthoquinone scaffolds have electron affinities within the CoQ0 substrate range, validating their quinone_A PAINS flags; the dimethoxybenzene CoQ_Mimic_001 (EA 1.26 eV) is redox-inert. Four PAINS-clean, Lipinski-compliant compounds were advanced as priority candidates. Conclusions. JKE-1674 and PipAryl_005 carry in silico dual GPX4–FSP1 and triple SLC7A11–FSP1 profiles against the experimental 8WIK receptor, both with better drug-likeness than iFSP1. FSP1 enzymatic assays (DCPIP) and SPR are the required next steps before either compound is described as a confirmed multi-pathway inhibitor.