title: Oxime-based linker libraries as a general approach for the rapid generation and screening of multidentate inhibitors.
abstract: The described oxime-based library protocol provides detailed procedures for the linkage of aminooxy functionality with aldehyde building blocks that result in the generation of libraries of multidentate inhibitors. Synthesis of inhibitors for protein tyrosine phosphatases (PTPs) and antagonists directed against the human tumor susceptibility gene 101 (TSG101) are shown as examples. Three steps are involved: (i) the design and synthesis of aminooxy platforms; (ii) tethering with aldehydes to form oxime-based linkages with sufficient purity; and (iii) direct in vitro biological evaluation of oxime products without purification. Each coupling reaction is (i) performed in capped microtubes at room temperature (20-23 °C); (ii) diluted for inhibitory evaluation; and (iii) screened with targets in microplates to provide IC(50) or K(d) values. The synthesis of the aminooxy platforms takes 3-5 d; tethering with the aldehydes takes 24 h; and inhibition assay of enzymes and protein-protein interactions takes 30 min and 2 h, respectively.
authors.affiliation: Chemical Biology Laboratory, Molecular Discovery Program, Center for Cancer Research, National Cancer Institute, US National Institutes of Health, National Cancer Institute-Frederick, Frederick, Maryland, USA.
authors.forename: Medhanit
authors.initials: M
authors.lastname: Bahta
authors.forename: Fa
authors.initials: F
authors.lastname: Liu
authors.forename: Sung-Eun
authors.initials: SE
authors.lastname: Kim
authors.forename: Andrew G
authors.initials: AG
authors.lastname: Stephen
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Fisher
authors.forename: Terrence R
authors.initials: TR
authors.lastname: Burke
mesh_terms: D000447:Aldehydes / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D055808:Drug Discovery / Q000379:methods*; D056827:Endosomal Sorting Complexes Required for Transport / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D006801:Humans; D010091:Oximes / Q000737:chemistry*; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors* / Q000737:chemistry; D054852:Small Molecule Libraries; D014157:Transcription Factors / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000737:chemistry; D000099177:Tsg101 Protein
chemical_list: D000447:Aldehydes; D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D010091:Oximes; D017027:Protein Tyrosine Phosphatases; D054852:Small Molecule Libraries; D014157:Transcription Factors; D054301:gag Gene Products, Human Immunodeficiency Virus; D000099177:Tsg101 Protein