O
HN
HN
O
Br
N
N
HN
N
N
N
O
O
S
N
H
N
H
NH
O
AQZ01
NH 2
N
N
HN
O
N
N
N
N
HN
N
Br
N
N
N
H
NH
N
O
Br-BX
CZ03
AQZ01
CZ11
HN
N
N
N
O
S
N
H
NH 2
N
N
N
CZ03
CZ11
Cl
Cl
NH 2
NH 2
N
N
N
N
N
N
NH 2
N
N
N
CZ22/23-dMB-PP1
N
CZ17/2-ClB-PP1
N
N
CZ23/34-dMB-PP1
N
N
N
N
N
N
CZ26/34-dClB-PP1
Cl
N
CZ49
N
N
N
N
N
N
N
N
O
O
CZ61
O
CZ62
O
CZ63
NH 2
N
N
O
CZ64
N
CZ25/25-dMB-PP1
Cl
N
N
N
N
N
N
NH 2
N
N
CZ50
NH 2
NH 2
N
N
N
CZ61
O
CZ62
O
CZ63
N
N
N
N
O
O
N
Cl
Cl
N
N
N
CZ49
CZ40
N
NH 2
N
N
N
Cl
NH 2
N
N
N
N
N
NH 2
N
N
N
Cl
NH 2
N
N
CZ26/34-dClB-PP1
CZ50
N
CZ24/35-dMB-PP1
N
Cl
NH 2
N
N
N
N
N
N
N
NH 2
NH 2
N
N
Cl
NH 2
N
N
Cl
NH 2
N
N
NH 2
N
N
N
CZ19/4-ClB-PP1
NH 2
CZ23/34-dMB-PP1
Cl
N
N
Cl
CZ22/23-dMB-PP1
N
CZ17/2-ClB-PP1
N
N
N
N
N
N
NH 2
N
NH 2
N
N
N
CZ15/4-MB-PP1
N
N
Cl
N
CZ40
N
NH 2
N
NH 2
N
N
N
N
N
NH 2
N
Cl
NH 2
NH 2
N
N
CZ25/25-dMB-PP1
Cl
Cl
NH 2
N
CZ13/2-MB-PP1
N
N
CZ24/35-dMB-PP1
Cl
N
N
NH 2
N
N
NH 2
N
CZ19/4-ClB-PP1
NH 2
N
N
N
N
N
NH 2
N
N
NH 2
N
N
CZ15/4-MB-PP1
CZ13/2-MB-PP1
Cl
NH 2
N
N
N
N
Br-BX
Cl
N
N
N
N
O
CZ64
Supplementary Figure 1: Chemical structures of the compounds screened in Figure 6.
N
Synthetic Methods
Materials obtained commercially were reagent grade and were used without further
purification. Reactions were monitored by thin layer chromatography (TLC) and/or
mass spectroscopy (LC-MS) using a Waters Acquity UPLC/ESI-TQD with an Acquity
UPLC. 1H NMR spectra were obtained on a Brucker 400 spectrometer at 400 MHz.
Synthesis of GXJ1-76
1-(tert-butyl)-3-(m-tolyloxy)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (GXJ1-76). To a
solution of 3-bromo-1-(tert-butyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (50mg,
0.186mmol), cesium carbonate (121.2mg, 0.372mmol), and pyridine (3.07µL,
0.038mmol) in DMF (0.9mL) was added m-cresol (23.3µL, 0.223mmol) and copper (I)
iodide (3.62mg, 0.019mmol). The reaction was heated to 140ºC for 24hrs, filtered, then
added to 20mL water and extracted with DCM. The combined organic layers were dried
with sodium sulfate, filtered and concentrated. The resulting oil was purified by reverse
phase HPLC to yield pure GXJ1-76 (1.9mg, 3.4% yield). 1H NMR (400MHz, DMSO-d6)
8.17 (1H, s), 7.28 (1H, t, J=7.85 Hz), 7.17 (1H, s), 7.13 (1H, d, J=8.16 Hz), 6.98 (1H,
d, J=7.43 Hz), 2.32 (3H, s), 1.65 (9H, s).
General procedure for the synthesis of AQZ01-AQZ06
In an oven-dried and argon-cooled disposable scintillation vial, 4-chloro-6,7dimethoxyquinazoline was dissolved in isopropanol. The appropriate substituted aniline
was then added and the reaction mixture was heated at reflux for at least 2 hours. The
reaction mixture was then cooled and the resulting solid collected by Büchner filtration
and washed with ice-cold isopropanol to yield the pure compound.
6,7-dimethoxy-N-phenylquinazolin-4-amine (AQZ01). 1H NMR (400MHz, DMSO-d6)
 11.31 (1H, br s), 8.82 (1H, s), 8.28 (1H, s), 7.69 (2H, d, J=7.55 Hz), 7.50 (2H, t,
J=7.85 Hz), 7.35 (1H, s), 4.03 (3H, s), 4.01 (3H, s).
N-(3-isopropylphenyl)-6,7-dimethoxyquinazolin-4-amine (AQZ02). 1H NMR
(400MHz, DMSO-d6) 11.28 (1H, br s), 8.81 (1H, s), 8.27 (1H, s), 7.52 (1H, dd, J=1.28
Hz, J=3.59 Hz), 7.41 (1H, t, J=7.85 Hz), 7.35 (1H, s), 7.22 (1H, d, J=7.67 Hz), 4.02 (3H,
s), 4.01 (3H, s), 1.53 (1H, s), 1.27 (3H, s), 1.25 (3H, s).
N-(3-(tert-butyl)phenyl)-6,7-dimethoxyquinazolin-4-amine (AQZ03). 1H NMR
(400MHz, DMSO-d6) 11.25 (1H, br s), 8.81 (1H, s), 8.25 (1H, s), 7.64 (1H, t, J=1.83
Hz), 7.53 (1H, m), 7.43 (1H, t, J=7.79 Hz), 7.34 (1H, s), 4.02 (3H, s), 4.01 (3H, s). 1.34
(9H, s).
6,7-dimethoxy-N-(naphthalene-1-yl)quinazolin-4-amine (AQZ04). 1H NMR (400MHz,
DMSO-d6) 11.67 (1H, br s), 8.65 (1H, s), 8.40 (1H, s), 8.05 (2H, m), 7.90 (1H, d,
J=8.40 Hz), 7.62 (3H, m), 7.36 (1H, s), 4.04 (3H, s), 4.03 (3H, s).
6,7-dimethoxy-N-(naphthalene-2-yl)quinazolin-4-amine (AQZ05). 1H NMR (400MHz,
DMSO-d6) 11.58 (1H, br s), 8.86 (1H, s), 8.40 (1H, s), 8.23 (1H, s), 8.03 (1H, d, J=8.89
Hz), 7.97 (2H, t, J=8.64 Hz), 7.87 (1H, dd, J=1.95 Hz, J=8.77 Hz), 7.57 (1H, m), 7.38
(1H, s), 4.05 (3H, s), 4.02 (3H, s).
N-(1H-indol-4-yl)-6,7-dimethoxyquinazolin-4-amine (AQZ06). 1H NMR (400MHz,
DMSO-d6) 11.36 (1H, br s), 8.68 (1H, s), 8.27 (1H, s), 7.46 (1H, d, J=8.04 Hz), 7.39
(1H, m), 7.34 (1H, s), 7.20 (1H, t, J=7.79 Hz), 7.13 (1H, d, J=7.06 Hz), 6.32 (1H, s), 4.02
(6H, s).
N-(3-bromophenyl)-6,7-dimethoxyquinazolin-4-amine (PD153035). 1H NMR
(400MHz, DMSO-d6) 11.36 (1H, br s), 8.89 (1H, s), 8.32 (1H, s), 8.03 (1H, s), 7.78
(1H, d, J=7.85 Hz), 7.46 (1H, t, J=7.97 Hz), 7.35 (1H, s), 4.03 (3H, s), 4.01 (3H, s).
Synthesis of 1
Scheme 1. Conditions: (a) formamidine acetate, EtOH, reflux, (b) acetic anhydride,
Et3N, DMAP, DCM, reflux, (c) POCl3, DIPEA, DCM, reflux, (d) 4-aminoindole, IPA, relux.
6-hydroxy-4-quinazolone (2). To a solution of 5-hydroxyanthranilic acid (500mg,
3.26mmol) in ethanol (20mL) was added formamidine acetate (510mg, 4.90mmol). The
reaction was heated under reflux until completed, as monitored by LCMS. The ethanol
was then removed by rotary evaporation and the solid product was filtered from water
as a grey solid (462.3mg, 85.7% yield). 1H NMR (400MHz, DMSO-d6) 7.90 (1H, s),
7.54 (1H, d, J=8.77 Hz), 7.41 (1H, d, J=2.80 Hz), 7.26 (1H, dd, J=2.92 Hz, J=8.77 Hz).
MS (ESI) m/z 163.76 (100%, 164.73 (20%) [M + H]+.
6-acetyl-4-quinazolone (3). To a solution of compound 2 (162mg, 1mmol) and
triethylamine (28µL, 0.2mmol) in dichloromethane (6.15mL) at 0ºC was added
dimethylaminopyridine (3.7mg, 0.03mmol). Acetic anhydride (283µL, 3.0mmol) was then
added dropwise. The reaction was then warmed to room temperature and monitored by
TLC until completed. The reaction was concentrated by rotary evaporation and the
resulting solid was purified by silica chromatography using a methanol/dichloromethane
gradient (0-10% MeOH/DCM) to yield pure 3 as a white solid (143.9mg, 70.6% yield).
1H
NMR (400MHz, DMSO-d6) 8.11 (1H, s), 7.83 (1H, d, J=2.43 Hz), 7.73 (1H, d, 8.77
Hz), 7.60 (1H, dd, J=2.68 Hz, J=8.77 Hz). MS (ESI) m/z 204.77 (100%), 205.41 (60%),
206.06 (100%), 206.71 (35%) [M + H]+.
6-acetyl-4-chloroquinazoline (4). To a solution of compound 3 (100mg, 0.49mmol) in
dichloromethane (3.5mL) at 0ºC was added diisopropylethylamine (214µL, 1.23mmol).
Phosphorus (V) oxychloride (54µL, 0.59mmol) was then added dropwise. The reaction
was allowed to warm to room temperature then heated to 45ºC under reflux. After
~24hours, the reaction was added to 25mL saturated sodium bicarbonate and extracted
with DCM. The combined organic layers were dried with Na2SO4, filtered, and
concentrated. The resulting oil was dissolved in 50% ethyl acetate/hexanes and
purified by silica chromatography (20-50% EtOAc/hexanes) to yield pure 4 as a white
solid (58.6mg, 53.9% yield). 1H NMR (400MHz, DMSO-d6) 9.13 (1H, s), 8.19 (1H, d,
J=9.13 Hz), 8.06 (1H, dd, J=0.43 Hz, J=2.50 Hz), 7.97 (1H, dd, J=2.50 Hz, J=9.07 Hz),
2.38 (3H, s). MS (ESI) m/z 222.78 (65%), 223.73 (90%), 225.34 (100%) [M + H]+.
4-((1H-indol-4-yl)amino)quinazolin-6-yl acetate (1). 4-aminoindole (25.9mg,
0.196mmol) was added to a solution of compound 4 (21.8mg, 0.098mmol) in methanol
(2mL) and the resulting solution was heated to 48ºC for 1hr. The solution was allowed to
cool to room temperature and then concentrated in vacuo. The resulting oil was
dissolved in 5% MeOH/DCM and purified by silica chromatography (2-5% MeOH/DCM)
to yield pure compound 1 as a pale yellow solid (5.1mg, 16.4% yield). 1H NMR
(400MHz, DMSO-d6) 11.17 (1H, s), 9.84 (1H, s), 8.42 (1H, s), 7.82 (1H, d, J=8.52 Hz),
7.66 (1H, d, J=8.77 Hz), 7.31 (1H, s), 7.17 (2H, m), 6.34 (1H, s), 2.38 (3H, s). MS (ESI)
m/z 318.48 (90%), 320.95 (100%), 321.94 (15%) [M + H]+.