title: An NMR-Guided Screening Method for Selective Fragment Docking and Synthesis of a Warhead Inhibitor.
abstract: Selective hits for the glutaredoxin ortholog of Brucella melitensis are determined using STD NMR and verified by trNOE and (15)N-HSQC titration. The most promising hit, RK207, was docked into the target molecule using a scoring function to compare simulated poses to experimental data. After elucidating possible poses, the hit was further optimized into the lead compound by extension with an electrophilic acrylamide warhead. We believe that focusing on selectivity in this early stage of drug discovery will limit cross-reactivity that might occur with the human ortholog as the lead compound is optimized. Kinetics studies revealed that lead compound 5 modified with an ester group results in higher reactivity than an acrylamide control; however, after modification this compound shows little selectivity for bacterial protein versus the human ortholog. In contrast, hydrolysis of compound 5 to the acid form results in a decrease in the activity of the compound. Together these results suggest that more optimization is warranted for this simple chemical scaffold, and opens the door for discovery of drugs targeted against glutaredoxin proteins-a heretofore untapped reservoir for antibiotic agents.
authors.affiliation: Department of Chemistry and Biochemistry, The University of Akron, Akron, OH 44325, USA. rbk11@zips.uakron.edu.
authors.forename: Ram B
authors.initials: RB
authors.lastname: Khattri
authors.affiliation: Department of Chemistry and Biochemistry, The University of Akron, Akron, OH 44325, USA. dlm93@zips.uakron.edu.
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Morris
authors.affiliation: Department of Chemistry and Biochemistry, The University of Akron, Akron, OH 44325, USA. cmd71@zips.uakron.edu.
authors.forename: Caroline M
authors.initials: CM
authors.lastname: Davis
authors.affiliation: Department of Pathology and Laboratory Medicine, University of North Carolina, Chapel Hill, NC 27599, USA. sbilinovich@gmail.com.
authors.forename: Stephanie M
authors.initials: SM
authors.lastname: Bilinovich
authors.affiliation: Department of Chemistry and Biochemistry, The University of Akron, Akron, OH 44325, USA. ajc127@zips.uakron.edu.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Caras
authors.affiliation: Department of Chemistry and Biochemistry, The University of Akron, Akron, OH 44325, USA. mjp9@uakron.edu.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Panzner
authors.affiliation: Department of Chemistry and Biochemistry, The University of Akron, Akron, OH 44325, USA. mad151@zips.uakron.edu.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Debord
authors.affiliation: Department of Chemistry and Biochemistry, The University of Akron, Akron, OH 44325, USA. tleeper@gmail.com.
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Leeper
mesh_terms: D001665:Binding Sites; D055808:Drug Discovery / Q000379:methods; D004353:Drug Evaluation, Preclinical; D006801:Humans; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D008968:Molecular Conformation; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D011485:Protein Binding; D011506:Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry*; D054852:Small Molecule Libraries; D013329:Structure-Activity Relationship
chemical_list: D008024:Ligands; D011506:Proteins; D054852:Small Molecule Libraries
keywords: FBDD; HSQC; STD; docking; glutaredoxin; ortholog; trNOE; warhead