Everolimus

Biological Activity

Everolimus (RAD001) also known as SDZ-RAD, Certican, Zortress and Afinitorm is an mTOR inhibitor with IC50 of 0.63 nM. Everolimus is an immunosuppressant analog of rapamycin (sirolimus) and inhibits growth factor-mediated proliferation of haematopoietic and nonhaematopoietic cells. Everolimus shows a potent inhibitory effect on posttransplant lymphoproliferative disorder (PTLD)-derived cells in vitro and in vivo.

Customer Product Validations & Biological Datas

	Source	Breast Cancer Res Treat (2015). Figure 2. Everolimus
	Method	flow cytometry
	Cell Lines	KPL-1 cell
	Concentrations	10 nM
	Incubation Time	5 d
	Results	Cells were exposed to everolimus at 10 nM for 5 days, and then flow cytometry was performed using Nim- DAPI staining. G0/G1 cell cycle arrest was observed in the two sensitive and the one intermediate cell line tested but was not observed in resistant lines

	Source	Breast Cancer Res Treat (2015). Figure 1. Everolimus
	Method	western blot
	Cell Lines	KPL-1 cell
	Concentrations	100 nM
	Incubation Time	10 min–48 h
	Results	We observed a significant inhibition of S6 phosphorylation in all tested cell lines after 30 min exposure to everolimus regardless of sensitivity to growth inhibitory effects.

Protocol

Cell Experiment
Cell lines	NCI-N87 and MKN45 cells
Preparation method	Proliferation assay. Gastric cancer cells (10,000 cells re-suspended in 1 ml medium) were seeded in 24-well plates. 24 h after seeding,0.5 ml medium containing RAD001 was added to yield desired concentrations of RAD001. At the time points indicated, gastric cancer cells were harvested and counted with a Coulter Z1 electronic cell counter (Beckman Coulter, UK).
Concentrations	2.5–160 n M
Incubation time	72 h

Animal Experiment
Animal models	NCI-N87 Tumor xenograft model with Pathogen-free, 4–6 week old, female CB-17 SCID mice
Formulation	saline
Dosages	5 mg/kg qd for 121 days
Administration	P.O

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

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 resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 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 resveratrol of 11.2 mg/kg.

Chemical Information

References

Everolimus and enteric-coated mycophenolate sodium ab initio after liver transplantation: midterm results.
Manzia et al. Transplant Proc. 2012 Sep;44(7):1942-5. PMID: 22974878.

Conversion from calcineurin inhibitors to everolimus with low-dose cyclosporine in renal transplant recipients with squamous cell carcinoma of the skin.
Caroti et al. Transplant Proc. 2012 Sep;44(7):1926-7. PMID: 22974873.

Everolimus and Erlotinib as Second- or Third-Line Therapy in Patients with Advanced Non-Small-Cell Lung Cancer.
Papadimitrakopoulou et al. J Thorac Oncol. 2012 Sep 10. PMID: 22968184.

Comparison of Outcomes in Patients With Versus Without Diabetes Mellitus After Revascularization With Everolimus- and Sirolimus-Eluting Stents (from the SORT OUT IV Trial).
Jensen et al. Am J Cardiol. 2012 Sep 4. PMID: 22959714.

Virtual Histology Assessment of Cardiac Allograft Vasculopathy Following Introduction of Everolimus-Results of a Multicenter Trial.
Arora et al. Am J Transplant. 2012 Sep 7. PMID: 22958738.

Mid-Term Results of Everolimus-Eluting Stent in a Japanese Population Compared With a US Randomized Cohort: SPIRIT III Japan Registry With Harmonization by Doing.
Saito et al. J Invasive Cardiol. 2012 Sep;24(9):444-50. PMID: 22954564.

A drug safety evaluation of everolimus in kidney transplantation.
Holdaas et al. Expert Opin Drug Saf. 2012 Sep 6. PMID: 22954349.