Abstract
The phosphatidylinositol-3-kinase (PI3K)-protein kinase B (Akt)-mammalian target of rapamycin (mTOR) signaling pathway is often constitutively activated in various human cancers, providing validated targets for cancer therapy. Among a series of 5-cyano-6-morpholino-4-substituted-pyrimidine analogs designed and synthesized based on PI3K target, 4-(2-(dimethylamino)vinyl)-2-(3-hydroxyphenyl)-6-morpholinopyrimidine-5-carbonitrile (WJD008) was selected for further pharmacological characterization because of its potent activity against PI3K signaling. WJD008 inhibited kinase activity of PI3Kα and mTOR with less activity against PIKK family members. In cellular settings, WJD008 abrogated insulin-like growth factor-I-activated PI3K-Akt-mTOR signaling cascade and blocked the membrane translocation of a pleckstrin homology domain containing enhanced green fluorescent protein-general receptor for phosphoinositides, isoform 1-pleckstrin homology fusion protein, suggesting down-regulation of phosphatidylinositol (3,4,5)-trisphosphate output induced by WJD008 resulted in inactivation of PI3K pathway. Consequently, WJD008 arrested cells in G1 phase without induction of apoptosis. Furthermore, WJD008 reversed the hyperactivation of the PI3K pathway caused by the oncogenic mutation of p110α H1047R and suppressed the proliferation and clonogenesis of transformed RK3E cells harboring this mutant. WJD008 was superior to the pan-PI3K inhibitor wortmannin against proliferation of a panel of cancer cells independently of their status of PI3K pathway or tissue originations. In summary, WJD008 is a potent dual PI3K/mTOR modulator with antiproliferative and anticlonogenic activity in tumor cells and transformed cells with PIK3CA mutant, which provides new clues for the design and development of this chemical scaffold as an anticancer drug.
Footnotes
This work was supported by the National Science and Technology Major Project Key New Drug Creation and Manufacturing Program [Grant 2009ZX09301-001]; the Knowledge Innovation Program of Chinese Academy of Sciences [Grant KSCX2-YW-R-25]; the National Natural Science Foundation of China [Grants 30721005, 90713034]; and the Science and Technology Commission of Shanghai Municipality Pujiang Talent Program (08PJ14114) [Grants 07dz05906, 09JC1416600].
Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
doi:10.1124/jpet.110.167940.
-
ABBREVIATIONS:
- PI3K
- phosphatidylinositol 3-kinase
- PTEN
- phosphatase and tensin homolog deleted from chromosome 10
- HR
- H1047R
- PIKK
- phosphatidylinositol 3-kinase-related protein kinase
- mTOR
- mammalian target of rapamycin
- Akt
- protein kinase B
- DMSO
- dimethyl sulfoxide
- CHO
- Chinese hamster ovary
- FBS
- fetal bovine serum
- p
- phosphorylated
- EGFP
- enhanced green fluorescent protein
- Grp1
- general receptor for phosphoinositides, isoform 1
- PH
- pleckstrin homology
- CHAPS
- 3-[(3-cholamidopropyl)-dimethyl-ammonio]-1-propanesulfonate
- ELISA
- enzyme-linked immunosorbent assay
- TOR
- target of rapamycin
- PBST
- phosphate-buffered saline containing 0.1% Tween 20
- OD
- optical density
- IGF
- insulin-like growth factor
- GAPDH
- glyceraldehyde-3-phosphate dehydrogenase
- SRB
- sulforhodamin B
- DNA-PK
- DNA-dependent protein kinase
- PKD1
- phosphatidylinositol 3-phosphate-dependent kinase 1
- ATM
- ataxia telangiectasia mutated
- ATR
- ataxia telangiectasia mutated- and Rad3-related
- LY294002
- 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
- CPT
- camptothecin
- 4E-BP1
- eukaryotic translation initiation factor 4E binding protein 1
- p70S6K
- the 70-kDa ribosomal S6 kinase
- GFP
- green fluorescent protein
- PtdIns(3,4,5)P3
- phosphatidylinositol (3,4,5)-trisphosphate
- NT
- nontransfected
- XL765
- N-(3-(N-(3-(3,5-dimethoxyphenylamino)quinoxalin-2-yl)sulfamoyl)phenyl)-3-methoxy-4-methylbenzamide
- SF1126
- 4-(17-carboxy-14-(carboxymethyl)-8-(3-guanidinopropyl)-18-hydroxy-3,6,9,12,15-pentaoxo-2-oxa-7,10,13,16-tetraazaoctadecyl)-4-(4-oxo-8-phenyl-4H-chromen-2-yl)morpholin-4-ium.
- Received March 22, 2010.
- Accepted June 2, 2010.
- Copyright © 2010 by The American Society for Pharmacology and Experimental Therapeutics
JPET articles become freely available 12 months after publication, and remain freely available for 5 years.Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page.
|