GDC-0349

Biological Activity

GDC-0349 is a potent small molecule mTOR inhibitor with Ki of 3.8 nM. GDC-0349 inhibits downstream markers of mTOR, including phospho-4EBP1 and phospho-Akt(S473) in an in vivo PK/PD study in mouse, consistent with an inhibition of both mTORC1 and mTORC2 complexes. GDC-0349 inhibits tumor growth in a dose-dependent manner when dosed orally once daily in athymic mice in a MCF7-neo/Her2 tumor xenograft model. GDC-0349 is also efficacious in other xenograft models, including PC3 (PTEN null) and 786-O (VHL mutant). GDC-0349 is currently in a Phase I clinical trials in the treatment with solid tumors or Non-Hodgkin's lymphoma.

Protocol

Cell Experiment
Cell lines
Preparation method	Compound activity is tested in 10 point dose curves starting at the highest final concentration of 10 uM. They are serially diluted in 100% DMSO prior to further dilution with assay buffer. The reaction mixture (8 uls) containing 0.25 nM mTOR+GBL enzyme, 400 nM GFP-4E-BP1, 8 uM ATP, 50 mM Hepes pH 7.5, 0.01% Tween 20, 10 mM MnCl2, 1 mM EGTA, 1 mM DTT, 1% DMSO (+/- compound) is incubated at room temperature for 30 minutes. 8 uL of solution containing 2 nM Tb-anti-p4E-BP1 antibody & 10 mM EDTA diluted TR-FRET buffer is then added and incubated for 30 minutes to stop the reaction. The plate is scanned with the Envision plate reader. Ki values are calculated in Assay Explorer using the Morrison ATP-competitive tight binding equation for Ki apparent determination.
Concentrations
Incubation time

Animal Experiment
Animal models	MCF7-neo/Her-2 mouse xenograft breast cancer model
Formulation	0.5% methylcellulose/0.2% Tween 80 (MCT)
Dosages	10, 20, 30, 40, 50, 60, 70, and 80 mg/kg QD for 24 days
Administration	orally

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

Discovery and Biological Profiling of Potent and Selective mTOR Inhibitor GDC-0349.
Pei Z, et al. ACS Med Chem Lett. 2012 Nov 29;4(1):103-7. PMID: 24900569.

Pyrimidoaminotropanes as potent, selective, and efficacious small molecule kinase inhibitors of the mammalian target of rapamycin (mTOR).
Estrada AA, et al. J Med Chem. 2013 Apr 11;56(7):3090-101. PMID: 23473235.