title: Bioinformatics-based discovery and characterization of an AKT-selective inhibitor 9-chloro-2-methylellipticinium acetate (CMEP) in breast cancer cells.
abstract: AKT is a promising target for anticancer drug development. In this work, a bioinformatics approach was applied to search for AKT inhibitors based on the correlation analysis between phospho-Ser473 AKT expression level and the antiproliferative data of NCI small molecule compounds against NCI 60 cancer cell lines, the candidate compounds were then subject to AKT kinase assay. The possible effects of potent compound on PI3K/AKT, PDK1, and MAPK, its antiproliferative and apoptosis-inducing effects on breast cancer cells which have high-levels of AKT activation were assessed by Western blot analysis, cell viability assay, and apoptosis assay. One compound, CMEP (NSC632855, 9-chloro-2-methylellipticinium acetate) was identified with all three correlation algorithm, Pearson's, Sperman's, and Kendall's, showing a high-ranked correlation coefficient. CMEP inhibits only AKT, but does not inhibit PI3K, PDK1, or MAPK. CMEP also inhibits heregulin-induced AKT activation, does not inhibit heregulin-induced MAPK activation in MCF-7 breast cancer cells. Increased concentrations of ATP reverse the AKT inhibitory effect of CMEP. CMEP inhibits growth and induces apoptosis in breast cancer cells which have high-levels of AKT activation and lack functional PTEN; however, CMEP only shows a minimal activity in NIH3T3 cells which do not have AKT activation. In conclusion, a lead compound CMEP, as an AKT selective inhibitor has been identified started with a bioinformatics-based approach. CMEP inhibits growth and induces apoptosis in cancer cells which have high-levels of AKT activation and lack PTEN or harbor PTEN mutation.
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, West Virginia University, 1 Medical Center Drive, Morgantown, WV 26506-9142, USA. mzhang2@hsc.wvu.edu
authors.forename: Manchao
authors.initials: M
authors.lastname: Zhang
authors.forename: Xueliang
authors.initials: X
authors.lastname: Fang
authors.forename: Hongpeng
authors.initials: H
authors.lastname: Liu
authors.forename: Ribo
authors.initials: R
authors.lastname: Guo
authors.forename: Xiaojin
authors.initials: X
authors.lastname: Wu
authors.forename: Bihua
authors.initials: B
authors.lastname: Li
authors.forename: Feng
authors.initials: F
authors.lastname: Zhu
authors.forename: Yan
authors.initials: Y
authors.lastname: Ling
authors.forename: Brian N
authors.initials: BN
authors.lastname: Griffith
authors.forename: Shaomeng
authors.initials: S
authors.lastname: Wang
authors.forename: Dajun
authors.initials: D
authors.lastname: Yang
mesh_terms: D000465:Algorithms; D017209:Apoptosis; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000201:enzymology* / Q000473:pathology; D045744:Cell Line, Tumor; D019295:Computational Biology; D004611:Ellipticines / Q000494:pharmacology*; D004789:Enzyme Activation; D004797:Enzyme-Linked Immunosorbent Assay; D005454:Fluorescence Polarization; D006801:Humans; D020287:In Situ Nick-End Labeling; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: C110955:9-chloro-2-methylellipticinium acetate; D004611:Ellipticines; D047428:Protein Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt