BEZ235

Biological Activity

BEZ235 (NVP-BEZ235) is a potent dual inhibitor of PI3K and mTOR. BEZ235 (NVP-BEZ235) is able to effectively and specifically block the dysfunctional activation of the PI3K pathway and induce G (1) arrest. It also inhibits the growth of human cancer in animal models. For Class I PI3K family, NVP-BEZ235 biochemical IC50 are 4nM against p110α, 75nM against p110β, 7nM against p110σ, 5nM against p110γ.

Product Citations

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  J Stroke Cerebrovasc Dis. 2023 Jan 16;32(3):106984.

  Inhibition of PI3K/Akt/mTOR signaling by NDRG2 contributes to neuronal apoptosis and autophagy in ischemic stroke
  BEZ235 purchased from AbMole

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  Cell Reports. 2019 Feb 5;1518-1532.e9.

  Pre-existing Functional Heterogeneity of Tumorigenic Compartment as the Origin of Chemoresistance in Pancreatic Tumors.
  BEZ235 purchased from AbMole

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  Cancer Biol Ther. 2018 May 25;1-20.

  Preclinical evaluation of novel PI3K/mTOR dual inhibitor SN202 as potential anti-renal cancer agent
  BEZ235 purchased from AbMole

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  Apoptosis. 2018 May 18.

  Down-regulating IL-6/GP130 targets improved the anti-tumor effects of 5-fluorouracil in colon cancer.
  BEZ235 purchased from AbMole

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  Cell Death and Disease. 2018 Jan 26;9:137-52.

  Downregulation of MCL-1 and upregulation of PUMA using mTOR inhibitors enhance antitumor efficacy of BH3 mimetics in triple-negative breast cancer
  BEZ235 purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Cancer Biol Ther (2018). Figure 3. BEZ235 (AbMole BioScience)
	Method	CCK-8 assay
	Cell Lines	RCC 786-0 cell line
	Concentrations	10 mmol/L
	Incubation Time	48 or 72 h
	Results	These IC50 values were obviously lower than that of the compounds BEZ235 and BKM120, which may be promising candidates for development as new drugs targeting the PI3K pathway.

	Source	Apoptosis (2018). Figure 6. NVP-BEZ235 (Abmole Bioscience. USA)
	Method	Western Blot
	Cell Lines	HT29 and SW480 cells
	Concentrations	96 nmol/l
	Incubation Time	24 h
	Results	Resembling the WB results from the BZA treatment, the inhibitors AG490, NVP-BEZ23 and AZD6244 imitated BZA function, down-regulating the pivotal protein of the JAK/STAT3, PI3K/AKT/mTOR and Ras/Raf/MEK/ERK signal pathways

	Source	Apoptosis (2018). Figure 5. NVP-BEZ235 (Abmole Bioscience. USA)
	Method	cell apoptosis assay
	Cell Lines	HT29 and SW480 cells
	Concentrations	96 nmol/l
	Incubation Time	24 h
	Results	In addition, using the respective AG490 (50 μmol/l), NVP-BEZ235 (96 nmol/l) and AZD6244 (32 μmol/l) inhibitors for 24 h to mimic the BZA effect in colon cancer cells, we observed that the Bad/Bcl-2 ratio was significantly up-regulated and that Ki-67 mRNA expression was substantially down-regulated in this treatment group compared with the control group

	Source	Cell Death & Disease (2018). Figure 6. BEZ235 (Abmole Bioscience)
	Method	IHC analysis
	Cell Lines	Tumor-bearing nude mice
	Concentrations	15 mg/kg
	Incubation Time	4 weeks
	Results	Following BEZ235 treatment with or without ABT263, the expression of MCL-1 was suppressed while PUMA was increased; correspondingly, the expressions of p-AKT and p-4EBP1 were significantly suppressed.

	Source	Cell Death & Disease (2018). Figure 5. BEZ235 (Abmole Bioscience)
	Method	oral gavage
	Cell Lines	Tumor-bearing nude mice
	Concentrations	15 mg/kg
	Incubation Time	4 weeks
	Results	Our results showed that combination therapy of BEZ235 and ABT263 significantly suppressed tumor growth, comparing with each drug treatment alone.

	Source	Cell Death & Disease (2018). Figure 4. BEZ235 (Abmole Bioscience)
	Method	CO-IP analysis
	Cell Lines	MDA-MB-231, MDA-MB-157, and MDA-MB-468 cells
	Concentrations	1 μM
	Incubation Time	24 h
	Results	BIM and BAK were obviously released to trigger apoptosis once MCL-1 was suppressed by adding BEZ235 in the presence of ABT-263. Of note, although PUMA was upregulated by BEZ235, less PUMA was bound to MCL-1 than control group due to inhibition of MCL-1.

	Source	Cell Death & Disease (2018). Figure 3. BEZ235 (Abmole Bioscience)
	Method	Immunoblotting analysis
	Cell Lines	MDA-MB-231, MDA-MB-157, and MDA-MB-468 cells
	Concentrations	1 μM
	Incubation Time	24 h
	Results	We found that, besides MCL-1, the expression of PUMA but not other members was significantly increased following treatment of BEZ235 with or without ABT263 in MDA-MB-231, MDA-MB-157, and MDA-MB-468 cells.

	Source	Cell Death & Disease (2018). Figure 2. BEZ235 (Abmole Bioscience)
	Method	colony formation assays
	Cell Lines	MDA-MB-231, MDA-MB-157, and MDA-MB-468 cells
	Concentrations	1 μM
	Incubation Time	24 h
	Results	The results showed that AZD8055 or BEZ235 in combination with ABT263 strongly inhibited cell proliferation in different TNBC cell lines.

	Source	Cell Death & Disease (2018). Figure 1. BEZ235 (Abmole Bioscience)
	Method	Immunoblotting analysis
	Cell Lines	TNBC cell lines
	Concentrations	1 μM
	Incubation Time	24 h
	Results	We found the mTOR inhibitors, especially the BEZ235 and AZD8055, could significantly inhibit the phosphorylation of AKT and 4EBP1 and efficiently led to decreasing MCL-1 expression.

	Source	Oncol Rep (2014). Figure 2. BEZ235
	Method	apoptosis assay
	Cell Lines	OCCC cells
	Concentrations	10 and 100 nM
	Incubation Time	72 h
	Results	OVISE cells were arrested at the G1 phase, but did not exhibit apoptosis (denoted by an increased proportion of cells in sub-G1), after 72 h of treatment with 10 and 100 nM NVP-BEZ235 (Fig. 2A).

	Source	Oncol Rep (2014). Figure 1. BEZ235
	Method	Western blot
	Cell Lines	OVISE cells
	Concentrations	10 and 100 nM
	Incubation Time	6 or 24 h
	Results	Treatment with NVP-BEZ235 suppressed pAKT expression, while treatment with temsirolimus did not. Similar results were observed in the KK cells.

Protocol (for reference only)

Cell Experiment
Cell lines	MKN45, BT474, SNU216 and NCI-N87 cell lines
Preparation method	Cell viability assay Cells were seeded at a density of 2000 cells per well in a 96-well plate and incubated overnight in complete medium. Cells were treated with either trastuzumab, BEZ235, Everolimus, AZD6244 alone, or trastuzumab combined with BEZ235 or Everolimus or AZD6244. After 72 h of incubation, cell viability was determined using the MTS tetrazolium substrate (CellTiter 96 Aqueous One Solution Cell Proliferation Assay, Promega, Madison, WI, USA) following the manufacturer’s instructions. The absorbance was measured at 490 nm using a spectrophotometer. All experiments were repeated three times with at least triplicate readings for each concentration.
Concentrations	0~800nM
Incubation time	72h

Animal Experiment
Animal models	Xenograft models in non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice
Formulation	BEZ235 was formulated in 0.9 % NaCl as a homogeneous suspension (9 mg/mL) and stored at 4 °C until further use in the in vivo experiments.
Dosages	45 mg/kg body weight, daily
Administration	oral gavage

Chemical Information

Molecular Weight	469.55
Formula	C30H23N5O
CAS Number	915019-65-7
Solubility (25°C)	DMSO 3 mg/mL
Storage	Powder          -20°C   3 years ;  4°C   2 years In solvent       -80°C   6 months ;  -20°C   1 month

Conversion of different model animals based on BSA (PMID: 27057123)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

References

[1] Civallero et al. Expert Opin Investig Drugs. NVP-BEZ235 alone and in combination in mantle cell lymphoma: an effective therapeutic strategy.

[2] Karar et al. Cancer Biol Ther. Dual PI3K/mTOR inhibitor NVP-BEZ235 suppresses hypoxia-inducible factor (HIF)-1α expression by blocking protein translation and increases cell death under hypoxia.

[3] Roberts et al. Clin Cancer Res. Combined PI3K/mTOR and MEK Inhibition Provides Broad Anti-Tumor Activity in Faithful Murine Cancer Models.

[4] Lin et al. J Chromatogr B Analyt Technol Biomed Life Sci. Determination of NVP-BEZ235, a dual PI3K and mTOR inhibitor, in human and mouse plasma and in mouse tissue homogenates by reversed-phase high-performance liquid chromatography with fluorescence detection.