AZD-9291 is an irreversible and mutant selective EGFR inhibitor with IC₅₀s of 12 and 1 nM against EGFR^L858R and EGFR^L858R/T790M, respectively.

IC50: 1 nM (EGFR^L858R/T790M), 12 nM (EGFR^L858R)^[1]

AZD-9291 (AZD9291) shows similar potency to early generation tyrosine kinase inhibitor (TKIs) in inhibiting EGFR phosphorylation in EGFR cells harboring sensitising EGFR mutants including PC-9 (ex19del), H3255 (L858R) and H1650 (ex19del), with mean IC₅₀ values ranging from 13 to 54 nM for AZD-9291. AZD-9291 also potently inhibits phosphorylation of EGFR in T790M mutant cell lines (H1975 (L858R/T790M), PC-9VanR (ex19del/T790M), with mean IC₅₀ potency less than 15 nM^[1].

The tumor-bearing mice are treated with AZD-9291 (5 mg/kg/day) for one to two weeks. Within days of treatment, 5 of 5 C/L858R mice displays nearly 80% reduction in tumor volume by magnetic resonance imaging MRI after therapy with AZD-9291, while 5 of 5 mice treated with vehicle shows tumor growth^[1]. AZD-9291 demonstrates improved rat PK, reduced hERG affinity, and improved IGF1R margins relative to the previously described compounds, and so this compound is selected for further investigation. AZD-9291 also offers an additional degree of broader chemical and profile diversity when compared to the previously described lead compounds. Upon dosing AZD-9291 in three efficacy models, The comparable efficacy is observed at relatively low doses (10 mg/kg per day). The excellent efficacy is also observed when AZD-9291 is dosed at 5 mg/kg per day^[2].

• [1]. Cross DA, et al. AZD9291, an irreversible EGFR TKI, overcomes T790M-mediated resistance to EGFR inhibitors in lung cancer. Cancer Discov. 2014 Sep;4(9):1046-61.

  [2]. Finlay MR, et al. Discovery of a potent and selective EGFR inhibitor (AZD9291) of both sensitizing and T790M resistancemutations that spares the wild type form of the receptor. J Med Chem. 2014 Oct 23;57(20):8249-67.

AZD-9291 is dissolved in DMSO and stored, and then diluted with appropriate medium before use^[1].

PC-9 cells are seeded into T75 flasks (5×10⁵ cells/flask) in RPMI growth media and incubated at 37°C, 5% CO₂. The following day the media is replaced with media supplemented with a concentration of EGFR inhibitor equal to the EC₅₀ concentration predetermined in PC-9 cells. Media changes are carried out every 2-3 days and resistant clones allowed to grow to 80% confluency prior to the cells being trypsinised and reseeded at the original seeding density in media containing twice the concentration of EGFR inhibitor. Dose escalations are continued until a final concentration of 1.5 μM Gefitinib, 1.5 μM Afatinib, 1.5 μM WZ4002 or 160 nM AZD-9291 are achieved^[1]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

AZD-9291 is suspended in 1% Polysorbate 80 (Mice)^[1].
AZD-9291 is suspended in 0.5% w/v HPMC/0.1% w/v Tween in deionized water at a concentration of 20 mg/mL (Rat)^[2].

Mice^[1]
The EGFR^L858R and EGFR^L858R+T790M mice (male and female) are used. AZD-9291 is suspended in 1% Polysorbate 80 and administered via oral gavage once daily at the doses of 7.5 mg/kg and 5 mg/kg, respectively. Mice are imaged weekly at the Vanderbilt University Institute of Imaging Science. For immunoblot analysis, mice are treated for eight hours with drug as described before dissection and flash freezing of the lungs. Lungs are pulverized in liquid nitrogen before lysis.
Rat^[2]
The male RccHan:WIST rats (10-week-old) are received a single oral dose of AZD-9291 (200 mg/kg). Blood glucose levels are measured using an Accuchek Active meter. Serum insulin concentrations are determined using a commercial rat ELISA kit. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Cross DA, et al. AZD9291, an irreversible EGFR TKI, overcomes T790M-mediated resistance to EGFR inhibitors in lung cancer. Cancer Discov. 2014 Sep;4(9):1046-61.

  [2]. Finlay MR, et al. Discovery of a potent and selective EGFR inhibitor (AZD9291) of both sensitizing and T790M resistancemutations that spares the wild type form of the receptor. J Med Chem. 2014 Oct 23;57(20):8249-67.

499.61

C₂₈H₃₃N₇O₂

1421373-65-0

Room temperature in continental US; may vary elsewhere

DMSO: ≥ 30 mg/mL

* "<1 mg/ml"="" means="" slightly="" soluble="" or="" insoluble.="" "≥"="" means="" soluble,="" but="" saturation="">