A-674563 is an orally active Akt inhibitor (Ki=11nM). A-674563 binds to the Akt ATP site and reduces phosphorylation of downstream targets including GSK3 and FOXO to inhibit tumor cell proliferation and survival. A-674563 maintains chondrocyte marker expression by inhibiting Sox9 degradation. A-674563 can be used for research on tumors and cartilage degeneration[1-4].
In vitro, A549, A427, NCI-H23, NCI-H358, NCI-H1975 and NCI-H1650 cells were treated with 0.03-0.70μM A-674563 for 1-72 hours. A-674563 reduced cell viability and proliferation, decreased G0/G1 phase cell proportion and increased S phase cell proportion[5]. A375, WM-115 and SK-Mel-2 cells were treated with 10-1000nM A-674563 for 30-72 hours. A-674563 inhibited cell proliferation and colony formation, increased dead cell number, early and late apoptotic cell proportion, and histone-DNA apoptotic signal, blocked Akt(Thr-308) and S6K1 phosphorylation[6]. U937 cells were treated with 10-500nM A-674563 for 12-72 hours. A-674563 dose-dependently inhibited cell viability and proliferation, induced cell death and reduced colony formation, activated caspase-3 and caspase-9 and promoted apoptosis[7].
In vivo, CX3CR1-overexpressing C57BL/6J mice received a single tail vein injection of 100mg/kg A-674563. One hour after the injection, the mice were exposed to hyperoxia for 24-72 hours. A-674563 decreased LC3-II/LC3-I ratio and CD11b expression, increased p62 expression, reduced LC3 co-localization within CD11b positive cells and CX3CR1 co-localization with CD11b, alleviated pulmonary fibrosis[8]. C57BL/6 mice received a single intravenous injection of 100mg/kg A-674563 1 hour before ischemia. The mice were then exposed to 67% H2 and 33% O2 for 4 hours of reperfusion. A-674563 increased infarct size percentage, elevated serum creatine kinase, creatine kinase MB isoenzyme and cardiac troponin I levels, decreased p-Akt1 expression[9]. Immunocompromised male SCID mice bearing 3T3-Akt1, MiaPaCa-2 and PC-3 xenograft tumors received oral 40mg/kg A-674563 for 20 days. A-674563 slowed tumor growth in xenograft tumors model, enhanced paclitaxel antitumor efficacy in the PC-3 model, caused body weight loss and increased plasma insulin[10].
References:[1] Kobayashi T, Fujita K, Kamatani T, et al. A-674563 increases chondrocyte marker expression in cultured chondrocytes by inhibiting Sox9 degradation. Biochem Biophys Res Commun. 2018 Jan 1;495(1):1468-1475.
[2] Hirai T, Wang W, Murono N, et al. Potential role of Akt in the regulation of fibroblast growth factor 21 by berberine. J Nat Med. 2024 Jan;78(1):169-179.
[3] Goßmann M, Linder P, Thomas U, et al. Integration of mechanical conditioning into a high throughput contractility assay for cardiac safety assessment. J Pharmacol Toxicol Methods. 2020 Sep;105:106892.
[4] Chen O, Cao Z, Li H, et al. High-concentration hydrogen protects mouse heart against ischemia/reperfusion injury through activation of thePI3K/Akt1 pathway. Sci Rep. 2017 Nov 1;7(1):14871.
[5] Chorner PM, Moorehead RA, et al. A-674563, a putative AKT1 inhibitor that also suppresses CDK2 activity, inhibits human NSCLC cell growth more effectively than the pan-AKT inhibitor, MK-2206. PLoS ONE. 2018 Feb 22;13(2):e0193344.
[6] Zou Y, Fan G, Wang X. Pre-clinical assessment of A-674563 as an anti-melanoma agent. Biochem Biophys Res Commun. 2016;473(4):1243-1250.
[7] Xu L, Zhang Y, Gao M, et al. Concurrent targeting Akt and sphingosine kinase 1 by A-674563 in acute myeloid leukemia cells. Biochem Biophys Res Commun. 2016 Apr 8;472(4):662-668.
[8] Chen Y, Zhang H, Li F, et al. Inhibition of CX3C receptor 1-mediated autophagy in macrophages alleviates pulmonary fibrosis in hyperoxic lung injury. Life Sci. 2020;259:118286.
[9] Chen O, Cao Z, Li H, et al. High-concentration hydrogen protects mouse heart against ischemia/reperfusion injury through activation of the PI3K/Akt1 pathway. Sci Rep. 2017 Nov 1;7:14871.
[10] Luo Y, Shoemaker AR, Liu X, et al. Potent and selective inhibitors of Akt kinases slow the progress of tumors in vivo. Mol Cancer Ther. 2005 Jun;4(6):977-86.