Design and Synthesis of Holu-7, a Quinazolinone Derivative With an Active Stereo Geometry and Polar...

Quinazolin-4-one scaffolds offer rich anticancer chemistry but often trade potency for selectivity. Building on a tubulin-targeting lead (MJ-65), 12 new derivatives (26-37) were synthesized to dissect geometry-activity relationships and enhance the polarity of C2-substituents. Cytotoxicity was profiled against cancerous (HCT116 and CAL27) and normal (FHC) cells. In silico studies employed CDOCKER against tubulin (PDB 5NJH), GROMACS for molecular dynamics (MD), and ProTox-3.0 for toxicity estimation. The previously synthesized geometry-matched pairs 21-25 were active at 10 μM. In contrast, their isomers 26-30 were largely inactive, revealing a configuration-dependent binding mechanism. Optimizing polarity yielded Holu-7 (compound 35; 2,4-dihydroxyphenyl at the C2-position), which exhibited sub-50 nM IC₅₀ in HCT116/CAL27 cells and > 10 μM in FHC cells and induced robust G₂/M arrest. Docking/MD supported stable vinca-site engagement, with hydrogen bonds to ASP179/ASN206 and π-stacking with TYR224. ProTox-3.0 predicted a higher LD₅₀ and absence of major organ toxicities relative to MJ-65. This data indicates that configuration-aware, polarity-tuned design can deliver selective quinazolinone anticancer candidates; therefore, Holu-7 merits further preclinical evaluation.