BEZ235

生物活性

BEZ235 (NVP-BEZ235, Dactolisib) ATP竞争性的抑制PI3K和mTOR，对p110α/γ/δ/β和mTOR(p70S6K)均有抑制作用，IC50分别为4 nM/5 nM/7 nM/75 nM/6 nM，对ATR也有抑制作用，IC50为21 nM；但对Akt和PDK1抑制作用微弱。BEZ235是ATP竞争性的PI3K和mTOR抑制剂 ，作用于p110α, p110γ, p110δ 和 p110β的IC50分别为4, 5, 7和75 nM。BEZ235 (NVP-BEZ235)也是mTORC1/2 催化剂的抑制剂。BEZ235明显降低mTOR激活激酶p70S6K的磷酸化水平。

实验操作 来自于公开的文献，仅供相同实验参考（如实验材料、目的不同，请参考其他文献）

细胞实验
细胞系	MKN45, BT474, SNU216 and NCI-N87 cell lines
方法	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.
浓度	0~800nM
处理时间	72h

动物实验
动物模型	Xenograft models in non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice
配制	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.
剂量	45 mg/kg body weight, daily
给药处理	oral gavage

不同实验动物依据体表面积的等效剂量转换表（数据来源于FDA指南）

	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，将白藜芦醇用于小鼠的剂量22.4 mg/kg 换算成大鼠的剂量，需要将22.4 mg/kg 乘以小鼠的K_m系数（3），再除以大鼠的K_m系数（6），得到白藜芦醇用于大鼠的等效剂量为11.2 mg/kg。

化学数据

储备液配制

以下数据基于产品分子量，对于特殊产品，请参照COA中的储备液配制条件和说明进行操作。

Concentration / Solvent Volume / Mass	1 mg	5 mg	10 mg
1 mM	2.1297 mL	10.6485 mL	21.297 mL
5 mM	0.4259 mL	2.1297 mL	4.2594 mL
10 mM	0.213 mL	1.0648 mL	2.1297 mL

使用AbMole产品发表的文献

• 

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

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

• 

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

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

• 

  Apoptosis. 2018 May 18.

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

• 

  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购买自AbMole

客户产品验证及相关生物数据

	数据来源	Cancer Biol Ther (2018). Figure 3. BEZ235 (AbMole BioScience)
	方法	CCK-8 assay
	细胞系	RCC 786-0 cell line
	浓度	10 mmol/L
	处理时间	48 or 72 h
	实验结果	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.

	数据来源	Apoptosis (2018). Figure 6. NVP-BEZ235 (Abmole Bioscience. USA)
	方法	Western Blot
	细胞系	HT29 and SW480 cells
	浓度	96 nmol/l
	处理时间	24 h
	实验结果	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

	数据来源	Apoptosis (2018). Figure 5. NVP-BEZ235 (Abmole Bioscience. USA)
	方法	cell apoptosis assay
	细胞系	HT29 and SW480 cells
	浓度	96 nmol/l
	处理时间	24 h
	实验结果	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

	数据来源	Cell Death & Disease (2018). Figure 6. BEZ235 (Abmole Bioscience)
	方法	IHC analysis
	细胞系	Tumor-bearing nude mice
	浓度	15 mg/kg
	处理时间	4 weeks
	实验结果	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.

	数据来源	Cell Death & Disease (2018). Figure 5. BEZ235 (Abmole Bioscience)
	方法	oral gavage
	细胞系	Tumor-bearing nude mice
	浓度	15 mg/kg
	处理时间	4 weeks
	实验结果	Our results showed that combination therapy of BEZ235 and ABT263 significantly suppressed tumor growth, comparing with each drug treatment alone.

	数据来源	Cell Death & Disease (2018). Figure 4. BEZ235 (Abmole Bioscience)
	方法	CO-IP analysis
	细胞系	MDA-MB-231, MDA-MB-157, and MDA-MB-468 cells
	浓度	1 μM
	处理时间	24 h
	实验结果	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.

	数据来源	Cell Death & Disease (2018). Figure 3. BEZ235 (Abmole Bioscience)
	方法	Immunoblotting analysis
	细胞系	MDA-MB-231, MDA-MB-157, and MDA-MB-468 cells
	浓度	1 μM
	处理时间	24 h
	实验结果	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.

	数据来源	Cell Death & Disease (2018). Figure 2. BEZ235 (Abmole Bioscience)
	方法	colony formation assays
	细胞系	MDA-MB-231, MDA-MB-157, and MDA-MB-468 cells
	浓度	1 μM
	处理时间	24 h
	实验结果	The results showed that AZD8055 or BEZ235 in combination with ABT263 strongly inhibited cell proliferation in different TNBC cell lines.

	数据来源	Cell Death & Disease (2018). Figure 1. BEZ235 (Abmole Bioscience)
	方法	Immunoblotting analysis
	细胞系	TNBC cell lines
	浓度	1 μM
	处理时间	24 h
	实验结果	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.

	数据来源	Oncol Rep (2014). Figure 2. BEZ235
	方法	apoptosis assay
	细胞系	OCCC cells
	浓度	10 and 100 nM
	处理时间	72 h
	实验结果	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).

	数据来源	Oncol Rep (2014). Figure 1. BEZ235
	方法	Western blot
	细胞系	OVISE cells
	浓度	10 and 100 nM
	处理时间	6 or 24 h
	实验结果	Treatment with NVP-BEZ235 suppressed pAKT expression, while treatment with temsirolimus did not. Similar results were observed in the KK cells.

参考文献

NVP-BEZ235 alone and in combination in mantle cell lymphoma: an effective therapeutic strategy.
Civallero et al. Expert Opin Investig Drugs. 2012 Aug 25. PMID: 22920938.

Dual PI3K/mTOR inhibitor NVP-BEZ235 suppresses hypoxia-inducible factor (HIF)-1α expression by blocking protein translation and increases cell death under hypoxia.
Karar et al. Cancer Biol Ther. 2012 Sep 1;13(11). PMID: 22895065.

Combined PI3K/mTOR and MEK Inhibition Provides Broad Anti-Tumor Activity in Faithful Murine Cancer Models.
Roberts et al. Clin Cancer Res. 2012 Aug 7. PMID: 22872574.

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.
Lin et al. J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Jul 15;901:9-17. PMID: 22727754.