MK-2206 2HCl

生物活性

MK-2206是一种高度选择性的Akt1/2/3抑制剂，IC50分别为8 nM/12 nM/65 nM；对250种其他蛋白激酶没有抑制活性。MK-2206抑制Akt的苏氨酸308位点和丝氨酸473位点的自身磷酸化作用。另外，MK-2206阻止Akt调节的下游信号分子（包括TSC2, PRAS40，及核糖体S6蛋白）的磷酸化作用。与抑制Ras 突变型细胞系(如NCI-H358, NCI-H23, NCI-H1299, 和Calu-6)相比，MK-2206 更有效地抑制Ras野生型细胞系(如A431, HCC827, 和NCI-H292)。MK-2206和细胞毒素药剂如erlotinib 和lapatinib联用作用于肺部NCI-H460肿瘤细胞或者卵巢A2780肿瘤细胞，MK-2206也显示出协同效应。

使用AbMole产品发表的文献

• Cell Rep. 2023 Apr 14;42(4):112378.

  AKT activity orchestrates marginal zone B cell development in mice and humans

• Cell Rep. 2023 Jun 27;42(7):112690.

  Identification of XAF1 as an endogenous AKT inhibitor

• 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

• 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

• 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

• 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

• 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

• BMC Biol. 2021 May 20;19(1):108.

  Very long intergenic non-coding (vlinc) RNAs directly regulate multiple genes in cis and trans

• 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

• 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

• Cell Biol Int. 2020 Mar 17.

  Fluid Shear Stress Induces Runx-2 Expression via Upregulation of PIEZO1 in MC3T3-E1 Cells

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

  Lateral Inhibition in Cell Specification Mediated by Mechanical Signals Modulating TAZ Activity.

• 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.

• 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.

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

  Inhibition of AKT suppresses the initiation and progression of BRCA1-associated mammary tumors.

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

  Tight Junction Proteins Claudin-1 and Occludin Are Important for Cutaneous Wound Healing

• Oncotarget. 2016 Feb 22.

  Mechanosensitive caveolin-1 activation-induced PI3K/Akt/mTOR signaling pathway promotes breast cancer motility, invadopodia formation and metastasis in vivo.

• 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.

• Cell Signal. 2015 Nov;2191-200.

  Discontinuing MEK inhibitors in tumor cells with an acquired resistance increases migration and invasion.

• 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.

• 2015 Apr.

  THE MECHANOBIOLOGICAL MECHANISMS OF CAVEOLAE/CAVEOLIN-1 IN LOW SHEAR STRESS-INDUCED BREAST CARCINOMA CELL MIGRATION AND INVASION

• 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.

• 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.

产品使用成果展示

	数据来源	Int J Biol Sci (2018 Oct). Figure 6. MK-2206 (Abmole Bioscience, Houston, USA)
	方法	oral
	细胞系/动物模型	mice
	浓度	240 mg/kg
	处理时间	-
	实验结果	We identified downstream pathways that might specifically affect the regulation of MK-2206 resistance or sensitivity.

	数据来源	Int J Biol Sci (2018 Oct). Figure 4. MK-2206 (Abmole Bioscience, Houston, USA)
	方法	oral
	细胞系/动物模型	mice
	浓度	240 mg/kg
	处理时间	-
	实验结果	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.

	数据来源	Int J Biol Sci (2018 Oct). Figure 5. MK-2206 (Abmole Bioscience, Houston, USA)
	方法	oral
	细胞系/动物模型	mice
	浓度	240 mg/kg
	处理时间	-
	实验结果	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.

	数据来源	Am J Pathol (2017). Figure 3. MK-2206 (Abmole Bioscience)
	方法	Western blot
	细胞系/动物模型	Human keratinocytes
	浓度	2 or 5 mmol/L
	处理时间	3 h
	实验结果	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).

	数据来源	J Cancer (2015). MK220- Figure 6. (AbMole Bio Science (Kowloon, Hongkong, China)
	方法
	细胞系/动物模型	In HepG2R cells
	浓度	2 μM
	处理时间
	实验结果	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.

	数据来源	J Cancer (2015). MK220- Figure 5. (AbMole Bio Science (Kowloon, Hongkong, China)
	方法
	细胞系/动物模型	HCC cell lines
	浓度	2 μM
	处理时间	72 h
	实验结果	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

	数据来源	J Cancer (2015). MK220- Figure 3. (AbMole Bio Science (Kowloon, Hongkong, China)
	方法	western blot
	细胞系/动物模型	HCC cell lines
	浓度	2 μM
	处理时间
	实验结果	"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)."

	数据来源	J Cancer (2015). MK220- Figure 2. (AbMole Bio Science (Kowloon, Hongkong, China)
	方法
	细胞系/动物模型	HCC cell lines
	浓度	2 μM
	处理时间	24 h
	实验结果	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.

	数据来源	J Cancer (2015). MK220- Figure 1. (AbMole Bio Science (Kowloon, Hongkong, China)
	方法
	细胞系/动物模型	HCC cell lines
	浓度
	处理时间	72 h
	实验结果	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.

	数据来源	Invest New Drugs (2014). MK-2206, Figure 5. (AbMole BioScience, Kowloon, Hongkong, China)
	方法
	细胞系/动物模型	EGI-1R and TFK-1R cells
	浓度
	处理时间	72 h
	实验结果	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.

	数据来源	Invest New Drugs (2014). MK-2206, Figure 4. (AbMole BioScience, Kowloon, Hongkong, China)
	方法
	细胞系/动物模型	TFK-1 SCR and TFK-1 AKT1/2 knock-down cells
	浓度	2 μM
	处理时间	72 h
	实验结果	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.

	数据来源	Invest New Drugs (2014). MK-2206, Figure 2. (AbMole BioScience, Kowloon, Hongkong, China)
	方法	Western blot
	细胞系/动物模型	CCA cell lines
	浓度	2 μM
	处理时间	24 h
	实验结果	MK-2206, AZD6244 and AZD8055 clearly suppress the phosphorylation of their respective targets or downstream effectors, i.e.

	数据来源	Invest New Drugs (2014). MK-2206, Figure 1. (AbMole BioScience, Kowloon, Hongkong, China)
	方法	MTT assay
	细胞系/动物模型	CCA cell lines
	浓度
	处理时间	72 h
	实验结果	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.

	数据来源	Biochem Pharmacol (2015). MK-2206, Figure 5. (Abmole Bioscience Kowloon, Hong Kong, China)
	方法	western blot
	细胞系/动物模型	CD4+ T cells
	浓度	20 mM
	处理时间	24 h
	实验结果	Similar results were obtained with AKT inhibitor MK-2206, which abolished the activation of AKT as well as mTORC1-mediated RPS6 phosphorylation.

	数据来源	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).
	方法	western blot
	细胞系/动物模型	MDA-MB-231 cells
	浓度	PI3K (10 µM LY294002), Akt (10 µM MK-2206) or mTOR (10 µM rapamycin)
	处理时间	1h
	实验结果	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.

	数据来源	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).
	方法	Cell motility assay
	细胞系/动物模型	MDA-MB-231 cells
	浓度	PI3K (10 μM LY294002), Akt (20 μM MK-2206), mTOR (10 μM rapamycin, Rap), FAK (10 μM PF573228) or Src (10 μM PP2)
	处理时间	1h
	实验结果	"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. "

	数据来源	Biochemical Pharmacology (2015). Figure 7. LY294002, MK-2206 and GSK1120212 were gifts from Abmole Bioscience (Kowloon, Hong Kong).
	方法	Western blot
	细胞系/动物模型	Purified CD4+ T cells
	浓度	20 mM LY294002, 10 mM MK-2206 as well as 10 mM GSK1120212
	处理时间	3 h
	实验结果	Arctigenin could inhibit mTORC1 activation via a way independent of PI3K/AKT and ERK.

	数据来源	Int J Cancer (2013). Figure 3.MK-2206 was obtained from AbMole BioScience (Kowloon, Hongkong)
	方法	Western blotting, Brdu proliferation assay,
	细胞系/动物模型	CCA cells
	浓度	1.7µM
	处理时间	24 or 72h
	实验结果	The novel AKT inhibitor MK-2206 augments the antiproliferative effect of RAD001 in CCA cell lines.

	数据来源	Invest New Drugs (2014). Figure 2.MK-2206 was obtained from AbMole BioScience (Kowloon, Hongkong)
	方法	Cell cycle analysis, Cell Death Detection ELISA, western blot
	细胞系/动物模型	EGI-1, Sk-ChA-1, TFK-1 cells
	浓度	2µM
	处理时间	24 or 48h
	实验结果	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.

	数据来源	Invest New Drugs (2014). Figure 1.MK-2206 was obtained from AbMole BioScience (Kowloon, Hongkong)
	方法	BrdU proliferation assay
	细胞系/动物模型	EGI-1, Sk-ChA-1, TFK-1 cells
	浓度	0,7,21,62,185,556,1667, 5000nM / 0, 6.3, 12.5, 25, 50, 75, 100nM
	处理时间	72h
	实验结果	Combined targeting of AKT and MEK as well as AKT and mTOR is highly synergistic in CCA cell lines.

	数据来源	Cellular Signalling 27 (2015) 2191–2200.Figure 4. MK-2206 was obtained from AbMole BioScience (Kowloon, HongKong).
	方法	Cell viability was analyzed by using Alamar blue assay(EGI-1).
	细胞系/动物模型	MDA-MB-231/MDA-MB-231R, A375/A375R
	浓度	2µM
	处理时间	72 hours
	实验结果	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.

	数据来源	Cellular Signalling 27 (2015) 2191–2200.Figure 1. MK-2206 was obtained from AbMole BioScience (Kowloon, HongKong).
	方法	Cell viability was analyzed by using MTT or Alamar blue assay(EGI-1).
	细胞系/动物模型	MDA-MB-231(HTB-26), EGI-1, HT-29, A549, HCT116, A375 and SW480
	浓度	5µM
	处理时间	72 hours
	实验结果	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.

实验参考

体外实验*
细胞系	CNE-1,CNE-2,HONE-1,SUNE-1
方法	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.
浓度	0,0.08,0.16,0.31,0.63,1.25,2.5,5,10μM
处理时间	72 or 96 hours

*上述方法来自公开文献，仅供相同目的实验参考。如实验目的、材料、方法不同，请参考其他文献。

体内实验*
动物模型	CNE-2 model in male BALB/c nude mice
配制	Formulated in 30% Captisol
剂量	MK-2206 (240 mg/kg, three times a week), MK-2206 (480 mg/kg, once a week), and 30% Captisol (Cydex) diluents
给药处理	Oral gavage for 2 weeks

*上述方法来自公开文献，仅供相同目的实验参考。如实验目的、材料、方法不同，请参考其他文献。

化学性质

分子量	480.39
分子式	C25H21N5O.2HCl
CAS号	1032350-13-2
溶解性（25°C）	DMSO 12 mg/mL
储存条件	粉末型式       -20°C   3年；4°C   2年 溶于溶剂       -80°C   6个月；-20°C   1个月
运输方式	冰袋运输，根据产品的不同，可能会有相应调整。

储备液配制

*下述溶液配置方法仅为基于分子量计算出的理论值。不同产品在配置溶液前，需考虑其在不同溶剂中的溶解度限制。

Concentration / Solvent Volume / Mass	1 mg	5 mg	10 mg
1 mM	2.0816 mL	10.4082 mL	20.8164 mL
5 mM	0.4163 mL	2.0816 mL	4.1633 mL
10 mM	0.2082 mL	1.0408 mL	2.0816 mL

不同实验动物依据体表面积的等效剂量转换表（参考来源于公开文献）

	小鼠	大鼠	兔	豚鼠	仓鼠	狗
重量 (kg)	0.02	0.15	1.8	0.4	0.08	10
体表面积 (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km 系数	3	6	12	8	5	20

动物 A (mg/kg) = 动物 B (mg/kg) ×	动物 B的Km系数
	动物 A的Km系数

例如，依据体表面积折算法，将化合物用于小鼠的剂量20 mg/kg 换算成大鼠的剂量，需要将20 mg/kg 乘以小鼠的K_m系数（3），再除以大鼠的K_m系数（6），得到化合物用于大鼠的等效剂量为10 mg/kg。

参考文献

[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.