Fig. 1: Discovery of KRAS(G12D) inhibitor and hit degrader.
a Chemical structure of a KRAS(G12D) degrader 2. A constructed ternary complex model of hit degrader 2 in complex with GDP-bound KRAS(G12D) and VHL. Inset on the right shows the interface of the drug-induced ternary complex. b Design and rational optimization of degrader. Relationship of chemical structure, degradation activity (DC50), and binding affinity to KRAS(G12D) or VHL. KRAS degradation activity was assessed by In-Cell ELISA assay in AsPC-1 cells treated with compounds for 24 h. Data are presented as the geometric mean (n = 3). Binding experiments were performed by KRAS(G12D)-immobilized or VHL/ElonginC/ElonginB(VCB)-immobilized SPR assay in multicycle or single-cycle kinetic format. KD values were calculated from fitted kinetic data (KD = kd/ka). Data are presented as the geometric mean (multicycle, n = 5; single-cycle, n = 3). Full details and representative sensorgrams for binary and ternary experiments were shown in Supplementary Fig. 2a, b. X-Ray crystal structure of ternary complex of ASP3082, GDP-bound KRAS(G12D) and VCB (PDB: 9L6F). The ternary complex structure (c), and insets depict interaction between VHL ligand and VHL protein (d), PPI interface of drug-induced ternary complex (e), and the 2Fo – Fc map for the ligand ASP3082 (1.5σ) in mesh (f). For X-ray crystallography data collection and refinement statistics, see Supplementary Table 7.
