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: Aditya Kulkarni, Phong Quang, Victoriana Curry, Renee Keyes, Weihong Zhou, Hyejin Cho, Jonathan Baffoe, Béla Török, Kimberly Stieglitz
• Journal: Chemical biology & drug design (Chem Biol Drug Des) Vol. 84 Issue 3 Pg. 270-81 (Sep 2014) ISSN: 1747-0285 [Electronic] England
• PMID: 24641677 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2014 John Wiley & Sons A/S.

Chemical References

• Enzyme Inhibitors
• Micelles
• Pyrazoles
• Pyrimidines
• pyrazolo(3,4-d)pyrimidine
• phospholipase D2
• Phospholipase D
• phospholipase D1

Topics

• Binding Sites
• Catalytic Domain
• Drug Design
• Enzyme Inhibitors (chemical synthesis, chemistry, metabolism)
• HEK293 Cells
• Humans
• Kinetics
• Micelles
• Molecular Dynamics Simulation
• Phospholipase D (antagonists & inhibitors, biosynthesis, genetics, metabolism)
• Protein Binding
• Pyrazoles (chemical synthesis, chemistry, metabolism)
• Pyrimidines (chemical synthesis, chemistry, metabolism)
• Structure-Activity Relationship