MK-2206 2HCl

Biological Activity

MK-2206 is a novel allosteric inhibitor of Akt with IC50 values of 8 nM, 12 nM and 65 nM for Akt1, Akt2 and Akt3, respectively. An Akt inhibitor may be particularly useful for cancers in which increased Akt signaling is associated with reduced sensitivity to cytotoxic agents or receptor tyrosine kinase inhibitors. In vitro, MK-2206 synergistically inhibited cell proliferation of human cancer cell lines in combination with molecular targeted agents such as erlotinib (an epidermal growth factor receptor inhibitor) or lapatinib (a dual epidermal growth factor receptor/human epidermal growth factor receptor 2 inhibitor). In vivo, MK-2206 in combination with these agents exerted significantly more potent tumor inhibitory activities than each agent in the monotherapy setting.

Product Citations

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  Cell Rep. 2023 Apr 14;42(4):112378.

  AKT activity orchestrates marginal zone B cell development in mice and humans
  MK-2206 2HCl purchased from AbMole

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  Cell Rep. 2023 Jun 27;42(7):112690.

  Identification of XAF1 as an endogenous AKT inhibitor
  MK-2206 2HCl purchased from AbMole

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  Front Immunol. 2023 Mar 15;14:1127552.

  Giardia duodenalis-induced G0/G1 intestinal epithelial cell cycle arrest and apoptosis involve activation of endoplasmic reticulum stress in vitro
  MK-2206 2HCl purchased from AbMole

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  PLoS Pathog. 2022 Sep 6;18(9):e1010808.

  White spot syndrome virus directly activates mTORC1 signaling to facilitate its replication via polymeric immunoglobulin receptor-mediated infection in shrimp
  MK-2206 2HCl purchased from AbMole

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  Sci Rep. 2022 Jul 6;12(1):11444.

  PD-L1 regulates cell proliferation and apoptosis in acute myeloid leukemia by activating PI3K-AKT signaling pathway
  MK-2206 2HCl purchased from AbMole

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  Anticancer Res. 2022 Jul;42(7):3325-3340.

  PI3K/Akt and Wnt/β-catenin Signaling Cross-regulate NF-κB Signaling in TNF-α-induced Human Lgr5+ Intestinal Stem Cells
  MK-2206 2HCl purchased from AbMole

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  Cells. 2021 Feb 18;10(2):430.

  Knockdown of AKT3 Activates HER2 and DDR Kinases in Bone-Seeking Breast Cancer Cells, Promotes Metastasis In Vivo and Attenuates the TGFβ/CTGF Axis
  MK-2206 2HCl purchased from AbMole

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  BMC Biol. 2021 May 20;19(1):108.

  Very long intergenic non-coding (vlinc) RNAs directly regulate multiple genes in cis and trans
  MK-2206 2HCl purchased from AbMole

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  Biochem Biophys Res Commun. 2021 Sep 10;569:1-9.

  Rapid tumor recurrence in a novel murine GBM surgical model is associated with Akt/PD-L1/vimentin signaling
  MK-2206 2HCl purchased from AbMole

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  Anticancer Res. 2021 May;41(5):2257-2275.

  Combined Targeting of AKT and mTOR Synergistically Inhibits Formation of Primary Colorectal Carcinoma Tumouroids In Vitro: A 3D Tumour Model for Pre-therapeutic Dгυg Screening
  MK-2206 2HCl purchased from AbMole

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  Cell Biol Int. 2020 Mar 17.

  Fluid Shear Stress Induces Runx-2 Expression via Upregulation of PIEZO1 in MC3T3-E1 Cells
  MK-2206 2HCl purchased from AbMole

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  Cell. 2019 Mar 7;176(6):1379-1392.e14.

  Lateral Inhibition in Cell Specification Mediated by Mechanical Signals Modulating TAZ Activity.
  MK-2206 2HCl purchased from AbMole

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  ACS Nano. 2019 Aug 27;13(8):8597-8608.

  Gadofullerene Nanoparticles Reverse Dysfunctions of Pancreas and Improve Hepatic Insulin Resistance for Type 2 Diabetes Mellitus Treatment.
  MK-2206 2HCl purchased from AbMole

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  Mol Med Rep. 2019 May;19(5):3519-3526.

  Overexpression of ribonuclease inhibitor induces autophagy in human colorectal cancer cells via the Akt/mTOR/ULK1 pathway.
  MK-2206 2HCl purchased from AbMole

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  Int J Biol Sci. 2018 Oct 3;14(13):1769-1781.

  Inhibition of AKT suppresses the initiation and progression of BRCA1-associated mammary tumors.
  MK-2206 2HCl purchased from AbMole

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  Am J Pathol. 2017 Jun;187(6):1301-1312.

  Tight Junction Proteins Claudin-1 and Occludin Are Important for Cutaneous Wound Healing
  MK-2206 2HCl purchased from AbMole

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  Oncotarget. 2016 Feb 22.

  Mechanosensitive caveolin-1 activation-induced PI3K/Akt/mTOR signaling pathway promotes breast cancer motility, invadopodia formation and metastasis in vivo.
  MK-2206 2HCl purchased from AbMole

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  Biochem Pharmacol. 2015 Aug 15;323-36.

  Arctigenin exerts anti-colitis efficacy through inhibiting the differentiation of Th1 and Th17 cells via an mTORC1-dependent pathway.
  MK-2206 2HCl purchased from AbMole

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  Cell Signal. 2015 Nov;2191-200.

  Discontinuing MEK inhibitors in tumor cells with an acquired resistance increases migration and invasion.
  MK-2206 2HCl purchased from AbMole

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  J Cancer. 2015 Sep 16;1195-205.

  Vertical Targeting of AKT and mTOR as Well as Dual Targeting of AKT and MEK Signaling Is Synergistic in Hepatocellular Carcinoma.
  MK-2206 2HCl purchased from AbMole

• 

  2015 Apr.

  THE MECHANOBIOLOGICAL MECHANISMS OF CAVEOLAE/CAVEOLIN-1 IN LOW SHEAR STRESS-INDUCED BREAST CARCINOMA CELL MIGRATION AND INVASION
  MK-2206 2HCl purchased from AbMole

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  Invest New Drugs. 2014 Dec;32(6):1144-54.

  Dual Inhibition of PI3K-AKT-mTOR- and RAF-MEK-ERK-signaling is synergistic in cholangiocarcinoma and reverses acquired resistance to MEK-inhibitors.
  MK-2206 2HCl purchased from AbMole

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  Int J Cancer. 2013 Nov;133(9):2065-76.

  Combined targeting of AKT and mTOR using MK-2206 and RAD001 is synergistic in the treatment of cholangiocarcinoma.
  MK-2206 2HCl purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Int J Biol Sci (2018 Oct). Figure 6. MK-2206 (Abmole Bioscience, Houston, USA)
	Method	oral
	Cell Lines	mice
	Concentrations	240 mg/kg
	Incubation Time	-
	Results	We identified downstream pathways that might specifically affect the regulation of MK-2206 resistance or sensitivity.

	Source	Int J Biol Sci (2018 Oct). Figure 4. MK-2206 (Abmole Bioscience, Houston, USA)
	Method	oral
	Cell Lines	mice
	Concentrations	240 mg/kg
	Incubation Time	-
	Results	Three weeks after the initiation of treatment, the volume of tumors in vehicle-treated mice had increased ~5.8-fold relative to baseline, whereas tumors in MK-2206-treated mice were significantly smaller, exhibiting only an approximate 2.8-fold increase.

	Source	Int J Biol Sci (2018 Oct). Figure 5. MK-2206 (Abmole Bioscience, Houston, USA)
	Method	oral
	Cell Lines	mice
	Concentrations	240 mg/kg
	Incubation Time	-
	Results	The overall RTVs (treated vs. non-treated tumors) for mice bearing Brca1-mutant tumor allografts were 60.8% and 62.6% for mice treated with MK-2206 and olaparib, respectively.

	Source	Am J Pathol (2017). Figure 3. MK-2206 (Abmole Bioscience)
	Method	Western blot
	Cell Lines	Human keratinocytes
	Concentrations	2 or 5 mmol/L
	Incubation Time	3 h
	Results	To investigate whether indeed decreased p-AKT activity can result in impaired scratch WH under our low calcium conditions we inhibited AKT phosphorylation by using MK-2206 (Figure 3E). This resulted in significantly impaired scratch WH in a dose-dependent manner (Figure 3F).

	Source	J Cancer (2015). MK220- Figure 6. (AbMole Bio Science (Kowloon, Hongkong, China)
	Method
	Cell Lines	In HepG2R cells
	Concentrations	2 μM
	Incubation Time
	Results	In HepG2R cells only the combination of MK-2206 and AZD8055 resulted in a stronger inhibition of proliferation, whereas the combination of AZD6244 with MK-2206, or AZD8055, even had strong antagonistic effects compared to MK-2206 or AZD8055 alone.

	Source	J Cancer (2015). MK220- Figure 5. (AbMole Bio Science (Kowloon, Hongkong, China)
	Method
	Cell Lines	HCC cell lines
	Concentrations	2 μM
	Incubation Time	72 h
	Results	As shown in Figure 5B, combined MK2206 or knockdown of AKT1/2 with AZD8055 results in an additional inhibition of proliferation compared to inhibition of mTOR or AKT alone

	Source	J Cancer (2015). MK220- Figure 3. (AbMole Bio Science (Kowloon, Hongkong, China)
	Method	western blot
	Cell Lines	HCC cell lines
	Concentrations	2 μM
	Incubation Time
	Results	"Neither the AKT inhibitor MK-2206, nor the mTOR inhibitor AZD8055 alone were able to significantly reduce the level of pGSK3β (S9) in any of the cell lines analyzed, even though both had a distinct impact on pAKT (S473)."

	Source	J Cancer (2015). MK220- Figure 2. (AbMole Bio Science (Kowloon, Hongkong, China)
	Method
	Cell Lines	HCC cell lines
	Concentrations	2 μM
	Incubation Time	24 h
	Results	The combination of MK-2206 and AZD8055, which has shown the most robust impact on proliferation, caused an almost complete reduction of cells in S and G2 phase in Hep3B and Huh-7.

	Source	J Cancer (2015). MK220- Figure 1. (AbMole Bio Science (Kowloon, Hongkong, China)
	Method
	Cell Lines	HCC cell lines
	Concentrations
	Incubation Time	72 h
	Results	Combined targeting of AKT and MEK synergistically inhibited proliferation of all HCC cell lines, although the synergistic effect appears less prominent in HepG2 cell line because of its high susceptibility to AZD6244.

	Source	Invest New Drugs (2014). MK-2206, Figure 5. (AbMole BioScience, Kowloon, Hongkong, China)
	Method
	Cell Lines	EGI-1R and TFK-1R cells
	Concentrations
	Incubation Time	72 h
	Results	Surprisingly, combining MEK and AKT, or MEK and mTOR inhibition was highly synergistic, and completely reversed the acquired resistance against AZD6244 in these cells. Furthermore, combining MK-2206 and AZD8055 was still highly effective in these cells.

	Source	Invest New Drugs (2014). MK-2206, Figure 4. (AbMole BioScience, Kowloon, Hongkong, China)
	Method
	Cell Lines	TFK-1 SCR and TFK-1 AKT1/2 knock-down cells
	Concentrations	2 μM
	Incubation Time	72 h
	Results	Synergistic effects of MK-2206 and AZD8055 were confirmed when AKT1/2 knockdown cells or control cells, were counted after incubationwith AZD8055, MK-2206, or the combination of both for 72 h, as indicated in Fig. 4.

	Source	Invest New Drugs (2014). MK-2206, Figure 2. (AbMole BioScience, Kowloon, Hongkong, China)
	Method	Western blot
	Cell Lines	CCA cell lines
	Concentrations	2 μM
	Incubation Time	24 h
	Results	MK-2206, AZD6244 and AZD8055 clearly suppress the phosphorylation of their respective targets or downstream effectors, i.e.

	Source	Invest New Drugs (2014). MK-2206, Figure 1. (AbMole BioScience, Kowloon, Hongkong, China)
	Method	MTT assay
	Cell Lines	CCA cell lines
	Concentrations
	Incubation Time	72 h
	Results	We then analyzed the effect of combined AKT and mTOR inhibition by combiningMK-2206 and AZD8055 (Fig. 1).We observed strong synergistic effects, with CI values below 0.3 in all cell lines tested.

	Source	Biochem Pharmacol (2015). MK-2206, Figure 5. (Abmole Bioscience Kowloon, Hong Kong, China)
	Method	western blot
	Cell Lines	CD4+ T cells
	Concentrations	20 mM
	Incubation Time	24 h
	Results	Similar results were obtained with AKT inhibitor MK-2206, which abolished the activation of AKT as well as mTORC1-mediated RPS6 phosphorylation.

	Source	Oncotarget (2016). Figure 11. Inhibitors of PI3K (LY294002), Akt (MK-2206), Src (PP2), FAK (PF573228) and mTOR (rapamycin) were purchased from Abmole Bioscience (Houston, TX, USA).
	Method	western blot
	Cell Lines	MDA-MB-231 cells
	Concentrations	PI3K (10 µM LY294002), Akt (10 µM MK-2206) or mTOR (10 µM rapamycin)
	Incubation Time	1h
	Results	As shown in Figure 11. Pretreating MDA-MB-231 cells with the specific inhibitors of PI3K (10 µM LY294002), Akt (10 µM MK-2206) and mTOR (10 µM rapamycin) completely abolished the LSS-induced MT1-MMP expression, indicating that the PI3K/Akt/mTOR pathway is required for LSS-induced MT1-MMP expression.

	Source	Oncotarget (2016). Figure 1. Inhibitors of PI3K (LY294002), Akt (MK-2206), Src (PP2), FAK (PF573228) and mTOR (rapamycin) were purchased from Abmole Bioscience (Houston, TX, USA).
	Method	Cell motility assay
	Cell Lines	MDA-MB-231 cells
	Concentrations	PI3K (10 μM LY294002), Akt (20 μM MK-2206), mTOR (10 μM rapamycin, Rap), FAK (10 μM PF573228) or Src (10 μM PP2)
	Incubation Time	1h
	Results	"Notably, inhibitors of PI3K (LY294002), Akt (MK-2206) and mTOR (rapamycin) markedly decreased the LSS-induced wound closure activity. However, pretreatment with inhibitors of FAK (PF573228) and Src (PP2) has no effect on the LSS-induced cell motility (Figure 1B). It is suggested that PI3K, Akt and mTOR might be participated LSS-induced cell motility in an FAK and Src-independent manner. "

	Source	Biochemical Pharmacology (2015). Figure 7. LY294002, MK-2206 and GSK1120212 were gifts from Abmole Bioscience (Kowloon, Hong Kong).
	Method	Western blot
	Cell Lines	Purified CD4+ T cells
	Concentrations	20 mM LY294002, 10 mM MK-2206 as well as 10 mM GSK1120212
	Incubation Time	3 h
	Results	Arctigenin could inhibit mTORC1 activation via a way independent of PI3K/AKT and ERK.

	Source	Int J Cancer (2013). Figure 3.MK-2206 was obtained from AbMole BioScience (Kowloon, Hongkong)
	Method	Western blotting, Brdu proliferation assay,
	Cell Lines	CCA cells
	Concentrations	1.7µM
	Incubation Time	24 or 72h
	Results	The novel AKT inhibitor MK-2206 augments the antiproliferative effect of RAD001 in CCA cell lines.

	Source	Invest New Drugs (2014). Figure 2.MK-2206 was obtained from AbMole BioScience (Kowloon, Hongkong)
	Method	Cell cycle analysis, Cell Death Detection ELISA, western blot
	Cell Lines	EGI-1, Sk-ChA-1, TFK-1 cells
	Concentrations	2µM
	Incubation Time	24 or 48h
	Results	Treating CCA cell lines with a combination of two inhibitors significantly increased the amount of cells in G1-phase compared to each respective compound alone. A significant induction of apoptosis was observed in TFK-1 cells for the treatment with AZD6244 in combination with either MK-2206 or AZD8055. MK-2206, AZD6244 and AZD8055 clearly suppress the phosphorylation of their respective targets or downstream effectors, i.e. AKT, ERK and pS6.

	Source	Invest New Drugs (2014). Figure 1.MK-2206 was obtained from AbMole BioScience (Kowloon, Hongkong)
	Method	BrdU proliferation assay
	Cell Lines	EGI-1, Sk-ChA-1, TFK-1 cells
	Concentrations	0,7,21,62,185,556,1667, 5000nM / 0, 6.3, 12.5, 25, 50, 75, 100nM
	Incubation Time	72h
	Results	Combined targeting of AKT and MEK as well as AKT and mTOR is highly synergistic in CCA cell lines.

	Source	Cellular Signalling 27 (2015) 2191–2200.Figure 4. MK-2206 was obtained from AbMole BioScience (Kowloon, HongKong).
	Method	Cell viability was analyzed by using Alamar blue assay(EGI-1).
	Cell Lines	MDA-MB-231/MDA-MB-231R, A375/A375R
	Concentrations	2µM
	Incubation Time	72 hours
	Results	Treatment of resistant cells with a combination of AKTi MK-2206 and mTORi AZD8055 was unable to suppress migration. However, adding low dose AZD6244 (500 nM) to the combination of MK-2206 and AZD8055 significantly suppresses both, migration and proliferation.

	Source	Cellular Signalling 27 (2015) 2191–2200.Figure 1. MK-2206 was obtained from AbMole BioScience (Kowloon, HongKong).
	Method	Cell viability was analyzed by using MTT or Alamar blue assay(EGI-1).
	Cell Lines	MDA-MB-231(HTB-26), EGI-1, HT-29, A549, HCT116, A375 and SW480
	Concentrations	5µM
	Incubation Time	72 hours
	Results	Acquired resistance in these cell lines was independent of AKT and mTOR signaling. However, both parental and resistant cell lines were highly susceptible to mTORi AZD8055 and even more to the combination of MK-2206 and AZD8055.

Protocol (for reference only)

Cell Experiment
Cell lines	CNE-1,CNE-2,HONE-1,SUNE-1
Preparation method	MK-2206 is dissolved in DMSO as a stock solution and diluted by culture media before use. Cells are seeded at an appropriate density per well in 96-well plates and incubated for 24 hours. Then MK-2206 (0~10μM) is added to the cells. Cell proliferation is determined after 72 or 96 hours.
Concentrations	0,0.08,0.16,0.31,0.63,1.25,2.5,5,10μM
Incubation time	72 or 96 hours

Animal Experiment
Animal models	CNE-2 model in male BALB/c nude mice
Formulation	Formulated in 30% Captisol
Dosages	MK-2206 (240 mg/kg, three times a week), MK-2206 (480 mg/kg, once a week), and 30% Captisol (Cydex) diluents
Administration	Oral gavage for 2 weeks

Chemical Information

Molecular Weight	480.39
Formula	C25H21N5O.2HCl
CAS Number	1032350-13-2
Solubility (25°C)	DMSO 12 mg/mL
Storage	Powder          -20°C   3 years ;  4°C   2 years In solvent       -80°C   6 months ;  -20°C   1 month

References

[1] Nishida et al. Hypertension. Phosphatidylinositol 3-Kinase/Akt Signaling Pathway Activates the WNK-OSR1/SPAK-NCC Phosphorylation Cascade in Hyperinsulinemic db/db Mice.

[2] Sangai et al. Clin Cancer Res. Biomarkers of Response to Akt Inhibitor MK-2206 in Breast Cancer.

[3] Pant et al. PLoS One. Inhibition of AKT with the Orally Active Allosteric AKT Inhibitor, MK-2206, Sensitizes Endometrial Cancer Cells to Progestin.

[4] Bressanin et al. Oncotarget. Harnessing the PI3K/Akt/mTOR pathway in T-cell acute lymphoblastic leukemia: Eliminating activity by targeting at different levels.

[5] Lai et al. Biochem J. A novel PKB/Akt inhibitor, MK-2206, effectively inhibits insulin-stimulated glucose metabolism and protein synthesis in isolated rat skeletal muscle.

[6] Simioni et al. Leukemia. Cytotoxic activity of the novel Akt inhibitor, MK-2206, in T-cell acute lymphoblastic leukemia.