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.
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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)
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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
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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
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