产品描述

Everolimus is a potent mTOR inhibitor that binds to FKBP-12. It is used alone or in combination with calcineurin inhibitors.

靶点活性

mTOR (FKBP12)

体外活性

Everolimus competes with immobilized FK 506 for binding to biotinylated FKBP12 (IC50: 0.12-1.8 nM) [1]. RAD001 inhibited proliferation in vitro (IC50 values<1 nM to >1 μM), and pS6 kinase and 4E-BP1 were inhibited. In vitro, RAD001 inhibited the proliferation of VEGF-stimulated and fibroblast growth factor-stimulated human endothelial cells [2]. Everolimus exhibited a dose-dependent inhibition in both the total cells and the stem cells from the BT474 cell line and the primary breast cancer cells. The IC50 values of everolimus for BT474 and the primary CSCs were 2,054 and 3,227 nM, or 29 times and 21 times greater than the IC50 values for their corresponding total cells, respectively [3].

体内活性

In vivo, in tumor models derived from either sensitive or insensitive cells, RAD001 reduced Tie-2 levels, the amount of mature and immature vessels, total plasma, and tumor VEGF. RAD001 did not affect blood vessel leakiness in normal vasculature acutely exposed to VEGF nor did it affect tumor vascular permeability[2]. Compared to the control group, the everolimus, trastuzumab, and drug combination groups showed significant reductions in mean tumor sizes. Compared to the mean xenograft tumor size in the trastuzumab group, the mean tumor size in the everolimus group was larger. When the two drugs were combined, the xenograft tumor size was smaller than those of the groups treated with everolimus or trastuzumab alone [3].

激酶实验

FKBP12 binding assay & Mixed lymphocyte reaction (MLR) : FKBP12 binding assay: Binding to the FK 506 binding protein (FKBP12) is indirectly assessed by means of an ELISA-type competition assay. FK 506 is included in each individual experiment as a standard, and the inhibitory activity is expressed as relative IC50 compared to FK 506 (rIC50 = IC50 Everolimus/IC50 FK 506). Mixed lymphocyte reaction (MLR): The immunosuppressive activities of RAP and its derivatives are assessed in a two-way MLR, using spleen cells of BALB/c and CBA mice. RAP is included in each individual experiment as a standard, and the inhibitory activity is expressed as relative IC50 compared to RAP (rIC50 = IC50 Everolimus/IC50 RAP).

细胞实验

The 3-(4-5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide dye reduction assay (MTT assay) is used to compare the effects of Everolimus or trastuzumab on total breast cancer cells and breast CSCs. The total cells and the stem cells from the BT474 cell line and the primary breast cancer cells are respectively seeded into 96-well plates with different concentrations of the drugs, with five wells for each concentration, and the cells are cultured at 37 °C with 5 % CO2 in an incubator for 24 hours. The concentrations of Everolimus are 1 nM, 10 nM, 100 nM, 1 μM and 10 μM, and the concentrations of trastuzumab are 0.5 μg/mL, 1 μg/mL, 10 μg/mL, 50 μg/mL, and 100 μg/mL. The combinatorial inhibitory effect of Everolimus and Trastuzumab on the in vitro growth of breast CSCs is examined by MTT assay as well using 10 μg/mL trastuzumab in combination of increasing concentrations of everolimus (1 nM, 10 nM, 100 nM and 1 μM). After drug treatment for 24 hours, 20 μL MTT [5 mg/mL in phosphate buffered saline (PBS)] is added to each well, and the cells are incubated at 37 °C with 5 % CO2 and saturated humidity for 4 hours. Following the subsequent removal of the supernatant, 150 μL dimethyl sulfoxide (DMSO) is added to each well, and the cells are vortexed for 10 minutes. The light absorbance (OD value) is measured for each well using an ELISA reader. Each experiment is repeated in triplicate, and dose–response curves are plotted. The probit software of the statistical software package SPSS 17.0 for Windows is used to calculate the inhibitory concentration (IC50) of Everolimus.(Only for Reference) Cell lines: BT474 cell line and the primary breast cancer cells

别名

依维莫司 , SDZ-RAD , RAD001

纯度

98%

分子量

958.22

分子式

C53H83NO14

CAS No.

159351-69-6

存储

Powder: -20°C for 3 years

In solvent: -80°C for 2 years

溶解度

Ethanol: 50 mg/mL

DMSO: 50 mg/mL

Water: Insoluble

( < 1 mg/ml refers to the product slightly soluble or insoluble )

参考文献

1. Sedrani R, et al. Chemical modification of rapamycin: the discovery of SDZ RAD. Transplant Proc. 1998 Aug;30(5):2192-4.

2. Lane HA, et al. mTOR inhibitor RAD001 (everolimus) has antiangiogenic/vascular properties distinct from a VEGFR tyrosine kinase inhibitor. Clin Cancer Res. 2009 Mar 1;15(5):1612-22.

3. Zhu Y, et al. Antitumor effect of the mTOR inhibitor everolimus in combination with trastuzumab on human breast cancer stem cells in vitro and in vivo. Tumour Biol. 2012 Oct;33(5):1349-62.

Note
For research use only.