Aminodeoxychorismate Synthase Inhibitors from One-Bead One-Compound Combinatorial Libraries:  “Staged” Inhibitor Design

• Published in: Journal of medicinal chemistry Vol. 49; no. 25; pp. 7413 - 7426
• Main Authors: Dixon, Seth, Ziebart, Kristin T, He, Ze, Jeddeloh, Melissa, Yoo, Choong Leol, Wang, Xiaobing, Lehman, Alan, Lam, Kit S, Toney, Michael D, Kurth, Mark J
• Format: Journal Article
• Language: English
• Published: WASHINGTON American Chemical Society 14.12.2006; Amer Chemical Soc
• Subjects: Acetates - chemical synthesis; Acetates - chemistry; Anti-Infective Agents - chemical synthesis; Anti-Infective Agents - chemistry; Benzoates - chemical synthesis; Benzoates - chemistry; Binding Sites; Biological and medical sciences; Carbon-Nitrogen Ligases - antagonists & inhibitors; Carbon-Nitrogen Ligases - chemistry; Cations, Divalent; Chemistry, Medicinal; Chorismic Acid - chemistry; Combinatorial Chemistry Techniques; Drug Design; Fluorescent Dyes; Hydroxybenzoate Ethers; Kinetics; Life Sciences & Biomedicine; Magnesium - chemistry; Medical sciences; Miscellaneous; Peptide Library; Peptides - chemical synthesis; Peptides - chemistry; Pharmacology & Pharmacy; Pharmacology. Drug treatments; Phenoxyacetates - chemical synthesis; Phenoxyacetates - chemistry; Protein Binding; Resins, Synthetic; Science & Technology; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Transaminases; POTENT; METABOLITES; SMALL-MOLECULE LIBRARIES; CHORISMATE-UTILIZING ENZYMES; ANTHRANILATE SYNTHASE; LIGAND; PEPTIDE LIBRARIES; IDENTIFICATION; MASS-SPECTROMETRY; ENCODING METHOD; Automation; Peptide library; Carbon-nitrogen ligases; Enzyme; Combinatorial chemistry; Enzyme inhibitor; Matrix assisted laser desorption ionization; Derivatization; Screening; Structure activity relation; Ligases; Fluorescent labelling; Time of flight method; Chemical compound library; Classification; Mass spectrometry
• ISSN: 0022-2623; 1520-4804
• DOI: 10.1021/jm0609869

4-Amino-4-deoxychorismate synthase (ADCS) catalyzes the first step in the conversion of chorismate into p-aminobenzoate, which is incorporated into folic acid. We aim to discover compounds that inhibit ADCS and serve as leads for a new class of antimicrobial compounds. This report presents (1) synthesis of a mass-tag encoded library based on a “staged” design, (2) massively parallel fluorescence-based on-bead screening, (3) rapid structural identification of hits, and (4) full kinetic analysis of ADCS. All inhibitors are competitive against chorismate and Mg2+. The most potent ADCS inhibitor identified has a K i of 360 μM. We show that the combinatorial diversity elements add substantial binding affinity by interacting with residues outside of but proximal to the active site. The methods presented here constitute a paradigm for inhibitor discovery through active site targeting, enabled by rapid library synthesis, facile massively parallel screening, and straightforward hit identification.