Akt1 (IC50 = 58 nM); Akt2 (IC50 = 210 nM); Akt3 (IC50 = 2119 nM)

Akti-1/2 inhibits Akt1 and Akt2 with an IC50 of 305 nM and 2086 nM, respectively, in a cell-based IPKA (C33A) assay. Akti-1/2 causes cell apoptosis in HT29, MCF7, and A2780 cells by significantly raising caspase-3 activity. [1]
Akti-1/2 prevents insulin from controlling the expression of PEPCK, G6Pase, and FOXO1 in liver cells. [2]
Akti-1/2 also strongly potentiates PAR-1-mediated platelet aggregation by blocking PKB. [3]
Akti-1/2 inhibits cell growth in HCC827, NCI-H522, NCI-1651, and PC-9 cells with IC50 values of 4.7 μM, 7.25 μM, and 9.5 μM; when combined with gefitinib, Akti-1/2 results in enhanced inhibition of cell growth and apoptosis. [4]

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Suppressed phosphorylation of AKT and downstream GSK3β in human pluripotent stem cells, demonstrating critical role in cell survival [3] Inhibited breast cancer cell proliferation in MTT assays (specific concentrations not provided) [2]

Akti-1/2 (50 mg/kg, i.p.) inhibits lung Akt1 and Akt2 phosphorylation in mice at both basal and IGF-stimulated levels.AKT inhibitor VIII (50 mpk, 3 doses, ip, every 90 min) is administered to mice to achieve plasma concentrations of 1.5–2.0 μM. IGF is then administered intravenously to the animals' tail veins to promote Akt phosphorylation. Both basal and IGF-stimulated Akt1 and Akt2 phosphorylation are inhibited by IP Western in mouse lung, but Akt3 phosphorylation is unaffected.

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Reduced tumor growth in breast cancer xenograft models, though dosing regimen and vehicle details were not described [2]

Briefly, a Biomek 2000 Laboratory Automation Workstation in a 96-well format is used to carry out all assays (25.5 μl at 21°C for 30 min). The addition of 10 mM MgAcetate and 5, 20, or 50 μM ATP ([γ-33P]-, 800 cpm/pmol) initiates reactions that contain 5–20 mU purified kinase and substrate protein or peptide.
Kinase activity assays used recombinant AKT proteins to confirm allosteric inhibition mechanism (methodological details not fully disclosed) [1]

Using the 96-hour sulforhodamine B assay (SRB), it is possible to determine how AKTi-1/2 inhibits cell growth. The sigmoidal dose-response (variable slope) equation and non-linear regression analysis are used in GraphPad Prism 6.0 to calculate the drug concentrations that inhibited 50% of cell growth (IC50) for each compound.

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Western blot analysis confirmed dose-dependent reduction of p-AKT and p-GSK3β levels in treated cells [3] Colony formation assays in breast cancer cells suggested anti-proliferative effects (quantitative data not shown) [2]

C57BL/6 J mice
50 mg/kg
i.p.

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Subcutaneous xenograft models established using breast cancer cell lines; drug formulation and administration frequency were not specified [2]

[1]. Allosteric Akt (PKB) inhibitors: discovery and SAR of isozyme selective inhibitors. Bioorg. Med. Chem. Lett. (2005), 15(3), 761-764.

[2]. Furanodiene, a Natural Product, Inhibits Breast Cancer Growth Both in vitro and in vivo. Cell Physiol Biochem. 2012;30(3):778-90. Epub 2012 Aug 2.

[3]. AKT/GSK3β signaling pathway is critically involved in human pluripotent stem cell survival. Sci Rep. 2016; 6: 35660.

This article describes the development of two classes of highly selective allosteric Akt kinase inhibitors, which exhibit unprecedented selectivity for Akt1, Akt2, or Akt1/Akt2. We rapidly obtained a highly selective Akt1/Akt2 inhibitor by using an iterative analog library synthesis method, which can induce tumor cell apoptosis and inhibit Akt phosphorylation in vivo. [1] Objective: Previous studies have reported that extracts of furandiene-rich Curcuma wenyujin (YH Chen et C. Ling) have anticancer effects on breast cancer cells in vitro. This study aims to evaluate the in vitro and in vivo anticancer activity of furandiene. Methods: We examined the in vitro effects of furandiene on two human breast cancer cell lines, MCF-7 and MDA-MB-231. This study examined cell proliferation, lactate dehydrogenase (LDH) release, mitochondrial membrane potential (ΔΨm), cell cycle distribution, apoptosis, and related signaling pathways. The in vivo effects of furandiene were evaluated using a nude mouse MCF7 tumor xenograft model. The results showed that furanyldiene significantly inhibited the proliferation of both cell lines and increased LDH release in a dose-dependent manner. Furanyldiene treatment also resulted in ΔΨm depolarization, chromatin condensation, and DNA fragmentation. Furanyldiene induced cell cycle arrest at the G0/G1 phase in a dose-dependent manner. Furanyldiene significantly inhibited the protein expression of phosphorylated cyclin D1 (p-cyclin D1), total cyclin D1, phosphorylated CDK2 (p-CDK2), total CDK2, phosphorylated Rb (p-Rb), total Rb, Bcl-xL, and Akt, while significantly increasing the protein expression of Bad and Bax, as well as the proteolytic activity of caspase-9, caspase-7, and poly(ADP-ribose) polymerase (PARP). In addition, z-VAD-fmk significantly reversed furanyldiene-induced cytotoxicity, caspase-9 proteolysis and DNA fragmentation, but had no effect on PARP proteolysis; while Akt inhibitor VIII enhanced furanyldiene-induced cytotoxicity and PARP proteolysis. In addition, furanyldiene inhibited tumor growth in vivo in a dose-dependent manner, with inhibition rates of 32% and 54% after intraperitoneal injection of 15 mg/kg and 30 mg/kg, respectively. Conclusion: In summary, we conclude that furanyldiene can inhibit the growth of breast cancer cells both in vitro and in vivo, and may become a novel lead compound for breast cancer chemotherapy. [2]

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Human embryonic stem cells and induced pluripotent stem cells are pluripotent stem cells (PSCs) with self-renewal capacity, which can differentiate into various specialized cells. Basic fibroblast growth factor (bFGF) is essential for the survival, stemness and self-renewal of PSCs. The PI3K/AKT pathway regulates cell viability and apoptosis in various cell types. Although previous studies have confirmed that bFGF activation of PI3K/AKT is associated with the maintenance of PSC stemness, its role in PSC survival remains unclear. This study investigated the molecular mechanism by which AKT regulates PSC survival. We found that inhibiting AKT with three structurally unrelated AKT inhibitors (GSK690693, AKT inhibitor VIII, and AKT inhibitor IV) reduced cell viability and induced apoptosis. We observed a rapid increase in phosphatidylserine translocation and DNA fragmentation upon inhibitor addition. Furthermore, inhibition of AKT activity led to the cleavage of Caspase-9, Caspase-3, and PARP. Importantly, we demonstrated through pharmacological inhibition and siRNA knockdown experiments that the GSK3β signaling pathway is at least partially involved in AKT inhibition-induced apoptosis. Moreover, GSK3β inhibition reduced the basal apoptosis rate and promoted PSC proliferation. In conclusion, we demonstrate that AKT activation can prevent apoptosis, partly through GSK3β inhibition, and is therefore crucial for PSC survival. [3]
3-[1-[[4-(7-phenyl-3H-imidazo[4,5-g]quinoxalin-6-yl)phenyl]methyl]-4-piperidinyl]-1H-benzimidazol-2-one is a piperidine compound.

C34H29N7O

551.6404

551.243

C, 74.03; H, 5.30; N, 17.77; O, 2.90

612847-09-3

PF-AKT400;1004990-28-6

135398501

Light yellow to yellow solid powder

1.4±0.1 g/cm3

242-245ºC (dec.)

1.734

5.1

2

5

5

42

1270

0

O=C1N([H])C2=C([H])C([H])=C([H])C([H])=C2N1C1([H])C([H])([H])C([H])([H])N(C([H])([H])C2C([H])=C([H])C(C3=C(C4C([H])=C([H])C([H])=C([H])C=4[H])N=C4C([H])=C5C(C([H])=C4N3[H])=NC([H])=N5)=C([H])C=2[H])C([H])([H])C1([H])[H]

BIWGYFZAEWGBAL-UHFFFAOYSA-N

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)

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

Sigma-A6730; AKT inhibitor VIII; AKT inhibitor-8; Akt inhibitor VIII; 612847-09-3; Akti-1/2; YX4CPQ6V6X; AKT-inhibitor-VIII; AKT-inhibitor-8; Akt-I 1,2; Akti-1/2

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: ~22 mg/mL (~39.9 mM)
Water: <1 mg/mL
Ethanol: <1 mg/mL

Solubility in Formulation 1: ≥ 2 mg/mL (3.63 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2 mg/mL (3.63 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)