Journal of Medicinal Chemistry
Drug Annotation
removed, and the aqueous portion was further extracted with DCM
(2 × 25 mL). The combined organics were dried over MgSO4, filtered,
and concentrated onto silica for purification. The crude product was
purified by flash silica chromatography, with elution gradient 0−4% 7 N
NH3/MeOH in DCM. Pure fractions were evaporated to dryness and
triturated with ether to afford N-[2-(2-dimethylaminoethylmethylamino)-
4-methoxy-5-[[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino]phenyl]prop-
2-enamide (75 mg, 39%) as a cream solid. 1H NMR (400 MHz, DMSO,
22 °C) δ 2.21 (6H, s), 2.29 (2H, t), 2.72 (3H, s), 2.89 (2H, t), 3.86 (3H,
s), 3.92 (3H, s), 5.77 (1H, dd), 6.27 (1H, dd), 6.43 (1H, dd), 7.04 (1H, s),
7.15 (1H, t), 7.2−7.27 (2H, m), 7.53 (1H, d), 7.91 (1H, s), 8.24 (1H, d),
8.33 (1H, d), 8.68 (1H, s), 9.14 (1H, s), 10.22 (1H, s); 13C NMR (176
MHz, DMSO, 22 °C) δ 32.8, 42.6, 45.1, 55.7, 56.0, 56.8, 105.3, 107.1,
110.4, 112.4, 113.3, 120.8, 121.2, 121.9, 125.3, 125.5, 125.9, 127.7, 132.4,
133.8, 137.3, 137.7, 145.8, 157.6, 158.9, 159.8, 161.5, 162.3. CHN analysis,
C28H33N7O2·H2O requires C = 64.97%, H = 6.82%, N = 18.94%. Found:
C = 64.6%, H = 6.7%, N = 18.8%. m/z (ES+) (M + H)+ = 500.42. HRMS:
(M + H)+ = C28H34N7O2 = 500.276 84, found 500.276 86.
AUTHOR INFORMATION
Corresponding Author
Notes
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The authors declare no competing financial interest.
ACKNOWLEDGMENTS
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The authors thank Didier Danillion, Gareth Hughes, Cath
Eberlein, and Scott Martin for expert technical assistance. We are
also grateful to Anne Galer and Paula Daunt for project team
leadership. We also acknowldege Susan Galbraith for her support
of the AZD9291 program. Jamie Scott, Jason Kettle, and Kevin
Foote are also gratefully acknowledged for their helpful
comments during the preparation of this manuscript.
ABBREVIATIONS USED
N-[2-(2-Dimethylaminoethylmethylamino)-4-methoxy-5-[[4-(1-
methylindol-3-yl)pyrimidin-2-yl]amino]phenyl]prop-2-enamide Me-
sylate Salt.
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EGFR, epidermal growth factor receptor; NSCLC, non-small-
cell lung cancer; TKI, tyrosine kinase inhibitor; GSH,
glutathione; PK, pharmacokinetics; WT, wild-type; AM,
activating mutant; DM, double mutant; LLE, ligand lipophilicity
efficiency; LCMS, liquid chromatography−mass spectrometry;
SAR, structure−activity relationship; HPLC, high-performance
liquid chromatography; DCM, dichloromethane; DMA, N,N-
dimethylacetamide; DMF, N,N-dimethylformamide; DIPEA,
N,N-diisopropylethylamine; DMSO, dimethyl sulfoxide;
TBAF, tetra-N-butylammonium fluoride; TFA, trifluoroacetic
acid; THF, tetrahydrofuran
To a stirred solution of N-[2-(2-dimethylaminoethylmethylamino)-4-
methoxy-5-[[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino]phenyl]-
prop-2-enamide (5g, 9.11 mmol) in acetone (45.5 mL) and water
(4.55 mL) at 50 °C was added methanesulfonic acid (0.893g, 9.11
mmol) as a solution in acetone (4.55 mL). The resulting mixture was
stirred for 1.5 h. The resulting solid was collected by filtration and
dried at 80 °C under vacuum to give the title salt (4.9 g, 94%) as a solid
(mp(DSC) = 246−259 °C). 1H NMR (400 MHz, acetone-d6, 22 °C) δ 2.72
(3H, s), 2.96 (3H, s), 3.01 (6H, s), 3.58 (3H, t), 3.87−3.90 (7H, m), 5.76
(1H, dd), 6.38−6.53 (2H, m), 7.12 (1H, t), 7.20 (1H, t), 7.29 (1H, s), 7.40
(2H, t), 8.07−8.16 (3H, m), 8.56 (1H, s), 9.30 (1H, s), 9.60 (1H, s), 9.66
(1H, s); 13C NMR (101 MHz, acetone-d6, 22 °C) δ 34.3, 39.9, 44, 45.5, 50.1,
55.2, 57, 105.9, 107.7, 112, 113.4, 123.4, 123.9, 124.4, 124.9, 125.1, 125.2,
126.9, 129.4, 131.5, 139.9, 140.2, 142.3, 144, 152.8, 153.7, 167.1, 169.3. CHN
analysis, C29H37O5N7S (0.1% w/w H2O) requires C = 58.41%, H = 6.27%,
N = 16.44%, S = 5.38%. Found: C = 58.6%, H = 6.3%, N = 16.5%, S = 5.2%.
m/z (ES+) (M + H)+ = 500.26. HRMS: (M + H)+ = C28H34N7O2 =
500.27684, found 500.2759.
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ASSOCIATED CONTENT
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S
* Supporting Information
Preparation and characterization for additional final compounds,
cell assay statistical analyses, pharmacodynamic assay procedure,
kinase selectivty data, binding mode computational modeling,
and enzyme/cell correlations. This material is available free of
R
dx.doi.org/10.1021/jm500973a | J. Med. Chem. XXXX, XXX, XXX−XXX