title: 1,3-disubstituted-4-aminopyrazolo [3, 4-d] pyrimidines, a new class of potent inhibitors for phospholipase D.
abstract: Phospholipase D enzymes cleave lipid substrates to produce phosphatidic acid, an important precursor for many essential cellular molecules. Phospholipase D is a target to modulate cancer-cell invasiveness. This study reports synthesis of a new class of phospholipase D inhibitors based on 1,3-disubstituted-4-amino-pyrazolopyrimidine core structure. These molecules were synthesized and used to perform initial screening for the inhibition of purified bacterial phospholipase D, which is highly homologous to the human PLD1 . Initially tested with the bacterial phospholipase D enzyme, then confirmed with the recombinant human PLD1 and PLD2 enzymes, the molecules presented here exhibited inhibition of phospholipase D activity (IC50 ) in the low-nanomolar to low-micromolar range with both monomeric substrate diC4 PC and phospholipid vesicles and micelles. The data strongly indicate that these inhibitory molecules directly block enzyme/vesicle substrate binding. Preliminary activity studies using recombinant human phospholipase Ds in in vivo cell assays measuring both transphosphatidylation and head-group cleavage indicate inhibition in the mid- to low-nanomolar range for these potent inhibitory novel molecules in a physiological environment.
authors.affiliation: Department of Chemistry, University of Massachusetts Boston, 100 Morrissey Blvd, Boston, MA, 02125, USA.
authors.forename: Aditya
authors.initials: A
authors.lastname: Kulkarni
authors.forename: Phong
authors.initials: P
authors.lastname: Quang
authors.forename: Victoriana
authors.initials: V
authors.lastname: Curry
authors.forename: Renee
authors.initials: R
authors.lastname: Keyes
authors.forename: Weihong
authors.initials: W
authors.lastname: Zhou
authors.forename: Hyejin
authors.initials: H
authors.lastname: Cho
authors.forename: Jonathan
authors.initials: J
authors.lastname: Baffoe
authors.forename: Béla
authors.initials: B
authors.lastname: Török
authors.forename: Kimberly
authors.initials: K
authors.lastname: Stieglitz
mesh_terms: D001665:Binding Sites; D020134:Catalytic Domain; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D008823:Micelles; D056004:Molecular Dynamics Simulation; D010739:Phospholipase D / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011720:Pyrazoles / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D011743:Pyrimidines / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D004791:Enzyme Inhibitors; D008823:Micelles; D011720:Pyrazoles; D011743:Pyrimidines; C014175:pyrazolo(3,4-d)pyrimidine; C105606:phospholipase D2; D010739:Phospholipase D; C106801:phospholipase D1
keywords: chemical biology; drug design; protease and ligands (substrate/inhibitor)