4-Hydroxyphenylpyruvate dioxygenase
(HPPD) is a functional protein
existing in almost all aerobic organisms. In the field of agricultural
chemicals, HPPD is acknowledged to be one of the crucial targets for
herbicides at present due to its unique bio-function in plants. In
the Auto Core Fragment in silico Screening (ACFIS)
web server, a potential HPPD inhibitor featuring 1,2,3-benzotriazine-4-one
was screened out via a pharmacophore-linked fragment virtual screening
(PFVS) method. Molecular simulation studies drove the process of “hit-to-lead”
optimization, and a family of 1,2,3-benzotriazine-4-one derivatives
was synthesized. Consequently, 6-(2-hydroxy-6-oxocyclohex-1-ene-1-carbonyl)-5-methyl-3-(2-methylbenzyl)benzo[d][1,2,3]triazin-4(3H)-one (15bu) was identified to be the best HPPD inhibitor (IC50 =
36 nM) among the 1,2,3-benzotriazine-4-one derivatives, which had
over 8-fold improvement of enzyme inhibition compared with the positive
control mesotrione (IC50 = 289 nM). Crystallography information
for the AtHPPD–15bu complex revealed
several important interactions of the ligand bound upon the target
protein, i.e., the bidentate chelating interaction of the triketone
motif with the metal ion of AtHPPD, a tight π–π
stacking interaction consisting of the1,2,3-benzotriazine-4-one moiety
and two benzene rings of Phe-424 and Phe-381, and the polydirectional
hydrophobic contacts consisting of the ortho-CH3-benzyl group of the core scaffold and some hydrophobic residues.
Furthermore, compound 15bu displayed 100% inhibition
against the five species of target weeds at the tested dosage, which
was comparable to the weed control of mesotrione. Collectively, the
fused 1,2,3-benzotriazine-4-one-triketone hybrid is a promising chemical
tool for the development of more potent HPPD inhibitors and provides
a valuable lead compound 15bu for herbicide innovation.