Structural insights into the potency and selectivity of covalent pan-FGFR inhibitors

Abstract

FIIN-2, TAS-120 (Futibatinib) and PRN1371 are highly potent pan-FGFR covalent inhibitors targeting the p-loop cysteine of FGFR proteins, of which TAS-120 and PRN1371 are currently in clinical trials. It is critical to analyze their target selectivity and their abilities to overcome gatekeeper mutations. In this study, we demonstrate that FIIN-2 and TAS-120 form covalent adducts with SRC, while PRN1371 does not. FIIN-2 and TAS-120 inhibit SRC and YES activities, while PRN1371 does not. Moreover, FIIN-2, TAS-120 and PRN1371 exhibit different potencies against different FGFR gatekeeper mutants. In addition, the co-crystal structures of SRC/FIIN-2, SRC/TAS-120 and FGFR4/PRN1371 complexes reveal structural basis for kinase targeting and gatekeeper mutations. Taken together, our study not only provides insight into the potency and selectivity of covalent pan-FGFR inhibitors, but also sheds light on the development of next-generation FGFR covalent inhibitors with high potency, high selectivity, and stronger ability to overcome gatekeeper mutations.

Characterization of the cholangiocarcinoma drug pemigatinib against FGFR gatekeeper mutants

“…Pemigatinib was redocked into FGFR1 to evaluate the reliability of this method. Then, pemigatinib was docked into FGFR2 (PDB ID: 6LVL) 28 , FGFR3 (PDB ID:7DHL) 29 and FGFR4 (PDB ID: 7F3M) 30 , respectively. Structural models predict that FGFR1-4 in complex with pemigatinib exhibit similar binding modes and similar interaction patterns (Supplementary Fig.…”

Discovery of 6-Formylpyridyl Urea Derivatives as Potent Reversible-Covalent Fibroblast Growth Factor Receptor 4 Inhibitors with Improved Anti-Hepatocellular Carcinoma Activity

“…All FGFR4 constructs were prepared as previously described. ,,− Briefly, the FGFR4 kinase domain (residues 445–753) was cloned into a modified pET28a vector with an N-terminal PreScission protease-cleavable 6xHis tag. The FGFR4 kinase domain (residues 442–802) with an N-terminal TEL (residues 1–183) was cloned into pMSCVpuro (Clontech).…”

“…All FGFR4 crystals were grown at 4 °C by hanging drop vapor diffusion with well solution (15–21% PEG-3350, 0.1 M Bis–Tris, pH 4.5, and 0.2 M Li 2 SO 4 ), then quickly transferred to mother liquor supplemented with 20% glycerol and flash-frozen in liquid nitrogen. FGFR4/ 8k , FGFR4/ 8z , FGFR4 V550L / 8z , and FGFR4 V550M / 8z crystal data sets were collected at the BL19U136 beamlines of Shanghai Synchrotron Radiation Facility (SSRF), while the FGFR4/ 8r crystal data set was collected using an in-house MicroMax-007 X-ray generator equipped with VariMax HR optics (Rigaku, Japan) and processed using the HKL3000. , All structures were solved by molecular replacement using PHENIX Phaser with FGFR4/ 10z (PDB ID: 7YBO) as the search model for FGFR4/ 8r and FGFR4/ 8z , FGFR4 V550L / 7v (PDB ID: 7WCW) for FGFR4 V550L / 8z , and FGFR4 V550M / 7v (PDB ID: 7WCX) for FGFR4 V550M / 8z . The ligands were modeled in PRODRG Web site and fitted by Phenix.…”

Design, Synthesis, and Biological Evaluation of 5-Formyl-pyrrolo[3,2-b]pyridine-3-carboxamides as New Selective, Potent, and Reversible-Covalent FGFR4 Inhibitors

“…All constructs were prepared as previously described. ,, Briefly, the FGFR4 kinase domain, containing an N-terminal 6xHis tag followed by a PreScission cleavage site, was cloned into pET28a vector. The N -terminal of TEL followed by FGFR4 kinase domain (TEL-FGFR4) constructs were cloned into pMSCVpuro (Clontech).…”