title: Discovery of a novel class of AKT pleckstrin homology domain inhibitors.
abstract: AKT, a phospholipid-binding serine/threonine kinase, is a key component of the phosphoinositide 3-kinase cell survival signaling pathway that is aberrantly activated in many human cancers. Many attempts have been made to inhibit AKT; however, selectivity remains to be achieved. We have developed a novel strategy to inhibit AKT by targeting the pleckstrin homology (PH) domain. Using in silico library screening and interactive molecular docking, we have identified a novel class of non-lipid-based compounds that bind selectively to the PH domain of AKT, with "in silico" calculated K(D) values ranging from 0.8 to 3.0 micromol/L. In order to determine the selectivity of these compounds for AKT, we used surface plasmon resonance to measure the binding characteristics of the compounds to the PH domains of AKT1, insulin receptor substrate-1, and 3-phosphoinositide-dependent protein kinase 1. There was excellent correlation between predicted in silico and measured in vitro K(D)s for binding to the PH domain of AKT, which were in the range 0.4 to 3.6 micromol/L. Some of the compounds exhibited PH domain-binding selectivity for AKT compared with insulin receptor substrate-1 and 3-phosphoinositide-dependent protein kinase 1. The compounds also inhibited AKT in cells, induced apoptosis, and inhibited cancer cell proliferation. In vivo, the lead compound failed to achieve the blood concentrations required to inhibit AKT in cells, most likely due to rapid metabolism and elimination, and did not show antitumor activity. These results show that these compounds are the first small molecules selectively targeting the PH domain of AKT.
authors.affiliation: College of Medicine, Arizona Cancer Center, University of Arizona, Tucson, AZ 85721-0038, USA.
authors.forename: Daruka
authors.initials: D
authors.lastname: Mahadevan
authors.forename: Garth
authors.initials: G
authors.lastname: Powis
authors.forename: Eugene A
authors.initials: EA
authors.lastname: Mash
authors.forename: Benjamin
authors.initials: B
authors.lastname: George
authors.forename: Vijay M
authors.initials: VM
authors.lastname: Gokhale
authors.forename: Shuxing
authors.initials: S
authors.lastname: Zhang
authors.forename: Kishore
authors.initials: K
authors.lastname: Shakalya
authors.forename: Lei
authors.initials: L
authors.lastname: Du-Cuny
authors.forename: Margareta
authors.initials: M
authors.lastname: Berggren
authors.forename: M Ahad
authors.initials: MA
authors.lastname: Ali
authors.forename: Umasish
authors.initials: U
authors.lastname: Jana
authors.forename: Nathan
authors.initials: N
authors.lastname: Ihle
authors.forename: Sylvestor
authors.initials: S
authors.lastname: Moses
authors.forename: Chloe
authors.initials: C
authors.lastname: Franklin
authors.forename: Satya
authors.initials: S
authors.lastname: Narayan
authors.forename: Nikhil
authors.initials: N
authors.lastname: Shirahatti
authors.forename: Emmanuelle J
authors.initials: EJ
authors.lastname: Meuillet
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000037:antagonists & inhibitors; D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology; D001667:Binding, Competitive / Q000187:drug effects; D015195:Drug Design; D004354:Drug Screening Assays, Antitumor; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D019073:HT29 Cells; D006801:Humans; D055504:Insulin Receptor Substrate Proteins; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D016415:Sequence Alignment; D013846:Thiepins / Q000138:chemical synthesis / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000970:Antineoplastic Agents; C526292:IRS1 protein, human; D055504:Insulin Receptor Substrate Proteins; C526295:Irs1 protein, mouse; D013846:Thiepins; D051057:Proto-Oncogene Proteins c-akt
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