Molecular Weight (MW)

551.64

Formula

C34H29N7O

CAS No.

612847-09-3

Storage

-20℃ for 3 years in powder form

-80℃ for 2 years in solvent

Solubility (In vitro)

DMSO: 22 mg/mL (39.9 mM)

Water: <1 mg/mL

Ethanol: <1 mg/mL

Other info

Chemical Name: 3-[1-[[4-(7-phenyl-3H-imidazo[4,5-g]quinoxalin-6-yl)phenyl]methyl]piperidin-4-yl]-1H-benzimidazol-2-one

InChi Key: BIWGYFZAEWGBAL-UHFFFAOYSA-N

InChi Code: InChI=1S/C34H29N7O/c42-34-39-26-8-4-5-9-31(26)41(34)25-14-16-40(17-15-25)20-22-10-12-24(13-11-22)33-32(23-6-2-1-3-7-23)37-29-18-27-28(36-21-35-27)19-30(29)38-33/h1-13,18-19,21,25H,14-17,20H2,(H,35,36)(H,39,42)

SMILES Code: O=C1N(C2CCN(CC3=CC=C(C4=NC5=CC6=C(N=CN6)C=C5N=C4C7=CC=CC=C7)C=C3)CC2)C8=CC=CC=C8N1

Synonym

Akt Inhibitor-VIII; AKT inhibitor VIII; AKT-inhibitor-VIII; AKT inhibitor-8; AKT-inhibitor-8; Akt-I 1,2; Akti-1/2. Sigma-A6730

In Vitro

Kinase Assay: Briefly, all assays (25.5 μl at 21°C for 30 min) are performed using a Biomek 2000 Laboratory Automation Workstation in a 96-well format. Reactions contains 5–20 mU purified kinase along with substrate peptide or protein and are initiated by the addition of 10 mM MgAcetate and 5, 20, or 50 μM ATP ([γ-33P]-ATP, 800 cpm/pmol).

Cell Assay: The cell growth inhibitory effect of AKTi-1/2 is studied using the 96 h sulforhodamine B assay (SRB). Drug concentrations that inhibited 50% of cell growth (IC50) are calculated for each compound in GraphPad Prism 6.0 using non-linear regression analysis and sigmoidal dose–response (variable slope) equation. Cell lines: HCC827, NCI-H522, NCI-1651, and PC-9 cells

In a cell-based IPKA (C33A) assay, Akti-1/2 inhibits Akt1 and Akt2 with IC50 of 305 nM and 2086 nM, respectively. In HT29, MCF7, and A2780 cells, Akti-1/2 induces cell apoptosis by dramatically increasing caspase-3 activity. In liver cells, Akti-1/2 blocks insulin regulation of PEPCK, G6Pase expression, and FOXO1 activity. Akti-1/2 also strongly potentiates PAR-1-mediated platelet aggregation by blocking PKB. In HCC827, NCI-H522, NCI-1651, and PC-9 cells, Akti-1/2 inhibits cell growth with IC50 of 4.7 μM, 7.25 μM, and 9.5 μM, when in combination with gefitinib, Akti-1/2 causes enhanced inhibition of cell growth and apoptosis.

In Vivo

In mice, Akti-1/2 (50 mg/kg, i.p.) inhibits basal and IGF-stimulated Akt1 and Akt2 phosphorylation in lung. Mice are dosed with AKT inhibitor VIII (50 mpk, 3 doses, ip, every 90 min) achieving plasma concentrations of 1.5-2.0 μM, and then the animals are tail vein injected with IGF to stimulate Akt phosphorylation. By IP Western, both basal and IGF stimulated Akt1 and Akt2 phosphorylation are inhibited in mouse lung, with no effect on Akt3 phosphorylation

Animal model

Formulation & Dosage

References

Bioorg Med Chem Lett. 2005 Feb 1;15(3):761-4; Diabetes. 2007 Sep;56(9):2218-27.