Synthesis of highly functionalized 2,4-diaminoquinazolines as anticancer and anti-HIV agents

Article abstract of DOI:10.1016/j.bmcl.2010.06.056

Novel polyhalo 2,4-diaminoquinazolines 3a-3d were prepared by reacting polyhaloisophthalonitriles with guanidine carbonate under solvent-free conditions and in the absence of a catalyst with good yields (74-95%). A series of highly functionalized 2,4-diaminoquinazolines 4-5 were then synthesized based on 3a-3c. The anticancer activities of compounds 3-5 were evaluated in vitro against human cell lines such as Skov-3, HL-60, A431, A549, and HepG-2. Some of the compounds showed excellent cytotoxic activity and 5a was found to be the most potent derivative, with an IC₅₀ value lower than 2.5 μg/mL against the five tumor cell lines, making it more active than cisplatin. Representative compounds were also preliminarily evaluated as HIV-1 inhibitors in vitro, and 3c showed the most potent anti-HIV-1 activity with EC₅₀ values of 0.6 and 1.6 μg/mL, and TI values of >59.6 and 66.6, respectively.

Full text of DOI:10.1016/j.bmcl.2010.06.056

Bioorganic & Medicinal Chemistry Letters 20 (2010) 4432–4435
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Synthesis of highly functionalized 2,4-diaminoquinazolines as anticancer
and anti-HIV agents
*
Sheng-Jiao Yan , Han Zheng , Chao Huang, Yu-Yun Yan, Jun Lin
Key Laboratory of Medicinal Chemistry for Natural Resources (Yunnan University), Ministry of Education, School of Chemical Science and Technology,
Yunnan University, Kunming 650091, PR China
a r t i c l e i n f o
a b s t r a c t
Article history:
Novel polyhalo 2,4-diaminoquinazolines 3a–3d were prepared by reacting polyhaloisophthalonitriles
with guanidine carbonate under solvent-free conditions and in the absence of a catalyst with good yields
(74–95%). A series of highly functionalized 2,4-diaminoquinazolines 4–5 were then synthesized based on
3a–3c. The anticancer activities of compounds 3–5 were evaluated in vitro against human cell lines such
as Skov-3, HL-60, A431, A549, and HepG-2. Some of the compounds showed excellent cytotoxic activity
Received 30 March 2010
Revised 21 May 2010
Accepted 9 June 2010
Available online 15 June 2010
and 5a was found to be the most potent derivative, with an IC50 value lower than 2.5 lg/mL against the
five tumor cell lines, making it more active than cisplatin. Representative compounds were also prelim-
inarily evaluated as HIV-1 inhibitors in vitro, and 3c showed the most potent anti-HIV-1 activity with
Keywords:
Polyhalo 2,4-diaminoquinazoline
Polyhaloisophthalonitrile
Solvent-free synthesis
Anticancer activity
EC50 values of 0.6 and 1.6
l
g/mL, and TI values of >59.6 and 66.6, respectively.
Ó 2010 Elsevier Ltd. All rights reserved.
Anti-HIV activity
Derivatives of 2,4-diaminoquinazoline are widely used as inhib-
itors of dihydrofolate reductase1 to treat opportunistic infections2 in
acquired immuno-deficiency syndrome (AIDS) patients and are
bioavailability is seen in some systems where fluorine substitution
leads to improved hydrolytic stability. Due to the unique effects of
F-substituents in pharmaceutical formulations, the use of fluori-
nated heterocyclic compounds as bioactive or functional molecules
has recently increased.
To explore the biological activity of this type of derivative, an
efficient and concise approach to constructing fluorine-containing
or highly functionalized 2,4-diaminoquinazolines is necessary. At
the same time, the halo-atoms of polyhalo 2,4-diaminoquinazoline
3 can be employed in nucleophilic substitution with nucleophiles.
The cyano group can also be modified into an amido or carboxyl
group, providing many opportunities for constructing more molec-
ular libraries for biological activity screening.
In this study, an efficient method for the rapid generation of li-
braries of 2,4-diaminoquinazolines 3–5 based on the results of
reacting polyhaloisophthalonitrile 1 with guanidine carbonate un-
der solvent-free conditions was developed. Initially, polyhaloi-
sophthalonitrile 1a was reacted with guanidine carbonate 2 in
different solvents, including acetonitrile, tetrahydrofuran (THF),
1,4-dioxane, and N,N-dimethylformamide (DMF), under different
conditions. It was found that the intact starting materials were
completely recovered when the reaction mixture was refluxed in
certain solvents like acetonitrile, THF, 1,4-dioxane, whereas other
solvents such as DMF gave unidentified products without forming
the desired product 3a. This may be because the limited solubility
of 2 in acetonitrile, THF or 1,4-dioxane may have depressed the
reaction, and the temperature was not high enough to initiate
known to be effective as antitumor agents,3 inhibitors of
a1-adren-
o-receptors,4 anti-bacterial drugs,5 anti-microbials,6 and so on.7
The potent bioactivity and low toxicity of these derivatives have
meant that they have received increasing attention from research-
ers.8 Several methods have been used to synthesize 2,4-diaminoqui-
nazolines, including substitution of 2-aminobenzonitrile reacted
with cyanamide, cyanoguanidine, chloroformamidine hydrochlo-
ride, and guanidine,9 or 2-fluorobenzonitrile condensation with
guanidine.8,10 However, these procedures usually have shortcom-
ings such as low yields and impractical conditions, and the introduc-
tion of multiple substituents into the 2,4-diaminoquinazoline ring
often requires multistep reactions and complex experimental pro-
cesses. To our knowledge, the availability of procedures to prepare
highly functionalized and diverse 2,4-diaminoquinazoles is limited.
Polyhaloisophthalonitriles, particularly polyfluoro isophthalo-
nitriles, have been widely used as organic agents.11 Fluorine incor-
poration into biologically active compounds can alter drug
metabolism or enzyme substrate recognition.12 The hydrophobic
nature of fluorinated compounds has also been cited for its ability
to improve transport across the blood–brain barrier. Improved oral
* Corresponding author. Tel./fax: +86 871 5033215.
E-mail address: linjun@ynu.edu.cn (J. Lin).
These authors contributed equally to this Letter.
0960-894X/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmcl.2010.06.056

S.-J. Yan et al. / Bioorg. Med. Chem. Lett. 20 (2010) 4432–4435
4433
Table 2
the reaction in the case of a solvent with a low boiling point like
Synthesis of 3 in solvent-free conditions under microwave irradiation
acetonitrile. While the ease of forming intramolecular H-bonding
between 2 with a polar aprotic solvent such as DMF may retard
further reaction of 1a with 2, on the other hand, a high tempera-
ture would cause the reaction to become complicated, resulting
in side reactions, for instance, decomposition of the substrates, or
the products, and hydrolysis. We fortuitously found that a sol-
vent-free process works well when the reaction is performed by
simply grinding 1a with an almost-equal amount of guanidine car-
bonate 2 (ratio of 1:1.3) under solvent-free conditions at ca. 110 °C
for 60 min (Scheme 1 and Table 1). Such a reaction produced 3a at
an 85% isolated yield (Table 1, entry 1).
In order to facilitate the operational program, accelerate the
reaction, and improve the accuracy of the temperature, microwave
irradiation (MW) was used to examine the practicality of the sol-
vent-free synthetic route (Table 1). To this end, a set of experi-
ments was carried out using polyhaloisophthalonitrile 1a and
guanidine carbonate 2 as model substrates. The mixture, composed
of a 1:1.3 ratio of 1a to 2, was subjected to microwave irradiation
in solvent-free conditions (Table 1, entries 2–10). The results show
that the optimum reaction conditions required to obtain the prod-
uct 3a are a temperature of 110 °C and irradiation for 10 min at a
maximum power of 200 W (Table 1, entry 6).
Entry
1
X
Y
R
3
MW
T (°C) Time (min) Yielda (%)
1
2
3
4
1a Cl Cl Cl
3a 200 W 110
3b 200 W 110
3c 200 W 110
10
10
10
10
89
92
95
74
1b
1c
F
F
Cl
F
F
F
1d Cl Cl NH2 3d 200 W 110
a
Isolated yield based on polyhaloisophthalonitrile 1.
X
NH2
N
NC
Nu
N
NH2
-
Y
X
Y
NH2
N
X=Cl, F
Y=Cl, F
t
NC
X
4
N
NH2
Nu NH2
N
NC
Nu
3a: X=Cl, Y=Cl
3b: X=F, Y=Cl
3c : X=F, Y=F
N
Y
NH2
5 Y=Cl, F
Scheme 3. Synthesis of highly functionalized 2,4-diaminoquinazoline 4–5.
The compounds 1b–1d were then reacted with guanidine car-
bonate under the same conditions (Scheme 2). The reactions took
only 10 min to complete at 110 °C with a maximum power of
200 W of MW, and gave moderate to good yields (Table 2, entries
1–4).13 The different structures of polyhaloisophthalonitrile had a
clear influence with regard to their reactivity and yield (Table 2,
entries 1–4).
The reactivity of 1 was increased by the electron-attracting
properties of the group on the aromatic ring. As such, 1c was gen-
erally the best substrate. Under the experimental conditions, the
reactivity ranking of the different structures of polyhaloisophthalo-
nitrile was 1c > 1b > 1a > 1d.
To construct the diverse molecular library of highly functional-
ized 2,4-diaminoquinazolines, polyhalo 2,4-diaminoquinazoline 3
was reacted with a number of nucleophiles such as alkylamines,
aryl amines, phenols, and mercaptans in DMF, at either room tem-
perature or 90 °C under basic conditions, in order to replace the
halo-atom with the nucleophile (Scheme 3 and Table 3). This type
of reaction features an SNAr mechanism and forms the compound
library 4–5 with good yields.14,15
In order to verify the structure of the highly functionalized 2,4-
diaminoquinazolines, 5b was selected as a representative com-
pound and characterized via X-ray crystallography, as shown in
Figure 1 (CCDC 760830).16
Cl
Cl
Cl
NH2
N
+
NC
Cl
CN
Cl
NC
Cl
NH2
Solvent-free
2-
CO3
H2N NH2
N
NH2
Cl
1a
2
2
3a
The novel 2,4-diaminoquinazolines 3–5 were evaluated for
in vitro cytotoxic activity against a series of human cells according
to a previously described method.17 The tumor cell lines included
ovarian carcinoma (Skov-3), myeloid leukemia (HL-60), epidermoid
carcinoma (A431), lung adenocarcinoma (A549), and laryngeal car-
Scheme 1. Synthesis of 3a under solvent-free conditions.
Table 1
Synthesis of 3a under solvent-free conditions
Entry
Power
T (°C)
Time (min)
Yielda (%)
1
2
3
4
5
6
7
8
9
Grinding
110
100
110
120
100
110
120
100
110
120
60
10
10
10
10
10
10
10
10
10
85
80
83
84
82
89
85
83
88
84
Table 3
MW/180 W
MW/180 W
MW/180 W
MW/200 W
MW/200 W
MW/200 W
MW/220 W
MW/220 W
MW/220 W
Synthesis of 2,4-diaminoquinazoline 4–5
Entry
3
X
Y
4
Nu
Time (h)
Yield (%)a
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
3b
3c
3b
3c
3b
3c
3b
3c
3b
3c
3b
3c
3b
3c
3b
3c
3b
3b
F
F
F
F
F
F
F
F
F
F
F
F
F
F
F
F
F
F
Cl
F
Cl
F
Cl
F
Cl
F
Cl
F
Cl
F
Cl
F
Cl
F
4a
4b
4c
4d
4e
4f
4g
4h
4i
PhNH
PhNH
10
10
10
10
10
10
10
10
10
10
10
20
30
15
10
30
30
15
61
64
70
69
87
89
82
88
83
85
85
90
60
78
79
82
82
73
p-MeOPhNH
p-ClPhNH
BnNH
BnNH
t-BuNH
t-BuNH
Et2N
10
a
Isolated yield based on polyhaloisophthalonitrile 1a.
4j
Et2N
5a
5b
5c
5d
5e
5f
n-BuNH
n-BuNH
BnNH
BnNH
PhO
BnO
PhS
n-PrS
X
X
Y
NH2
N
+
NC
R
CN
X
NC
R
NH2
Solvent-free
2-
CO3
H2N NH2
N
NH2
MW: 200W
110 ºC
Y
1
2
3
2
X=Cl, F; Y=Cl, F
R=Cl, F, NH2
X=Cl, F; Y=Cl, F
R=Cl, F, NH2
Cl
Cl
5g
5h
a
Isolated yield based on polyhalo 2,4-diaminoquinazoline 3.
Scheme 2. Synthesis of compounds 3a–3d by solvent-free reaction.

4434
S.-J. Yan et al. / Bioorg. Med. Chem. Lett. 20 (2010) 4432–4435
X
NH2
N
NC
R
F
NH2
N
NC
F
N
NH2
NH NH2
N
Y
N
NH2
NC
X = F > Cl > alkylamino >
arylamino
Cl
3b
N
H
N
Cl
5a
NH2
Y = Cl > F
R = F > Cl > alkylamino
>mercaptan >aryloxyl
more active compounds:
Figure 2. Structure–activity relationship of 2,4-diaminoquinazolines.
Figure 1. X-ray crystal structure of 5b.
We found that substitution at the 5- and 7-positions with n-
BuNH, and at the 8-position with a chlorine atom, also plays an
important role in regulating cytotoxic activity. This was evident
when we observed that 5a and 5b showed good activity against
the tumor cells used in our study (Table 4, entries 15–16 and
Fig. 2).
The representative compounds 3a–3c, 4a–4b and 5g–5h were
subsequently evaluated for their inhibitory activity against HIV-1
replication in acutely infected C8166 cells in vitro, according to
procedures described in the literature.18 AZT was used as the refer-
ence compound. The results of the cytotoxicity studies are summa-
rized in Table 5. The seven target compounds exhibited moderate
cinoma (HepG-2) cells. Cisplatin (DDP) was used as a positive con-
trol. The assay results are shown in Table 4 (where the IC50 value
is defined as the concentration of a compound that corresponds to
50% growth inhibition). As shown in Table 4, some of the compounds
showed good activity against the tumor cells. In fact, 3b was found to
be more active than cisplatin against Skov-3, HL-60, and HepG-2
cells. In addition, the derivative 5a was more active than cisplatin
against HL-60, A549 and HepG-2 cells.
Comparison of the activities of the polyhalo 2,4-diaminoquinaz-
olines 3–4 suggested that substitution at the 5- and 7-positions
with fluorine atoms, and at the 8-position with a chlorine atom,
plays a vital role in the modulation of cytotoxic activity. This would
explain why 3b showed excellent activity against the five tumor
cells (Fig. 2 and Table 4, entry 2). Significantly, 3b was up to 35
times more active than cisplatin against HL-60 cells (Table 4, entry
2 vs 23).
The polyhalo 2,4-diaminoquinazolines 3–4 showed poor activity
against the tumor cell line A549 (Table 4, entries 1–14), and moder-
ate or good activity against the Skov-3, HL-60, and A431 cells.
In addition, with the exception of 4c, 4f and 4h, these com-
pounds were found to be more active against HepG-2 cells than
cisplatin (Table 4, entries 1–14 and Fig. 2).
With the exception of 5a and 5b, highly functionalized 2,4-dia-
minoquinazolines 5 showed poor or moderate activity against
A549, A431, and HepG-2 cells and good activity against Skov-3
and HL-60 cells (Table 4, entries 15–22).
activity against HIV-1IIIB with EC50 values of 0.1–17.3 lg/mL and
TI values of 7.0–66.6 (Table 5, entries 1–7). The number of fluorine
atoms in the compounds influenced the activity and the toxicity of
the tested compounds. The TI values of the corresponding com-
pounds increased sharply (3c vs 3b and 3a; 4b vs 4a) as the num-
ber of fluoro atoms increased. Thus the TI values were ranked as
3c > 3b > 3a (entries 1–3), 4b > 4a (entries 4 and 5), and 4g ꢀ 4h
(entries 6 and 7). Notably, 3c was found to have the most potent
anti-HIV-1 activity with EC50 values of 0.6/1.6 lg/mL and
TI = 59.6/66.6, respectively (entry 3). Therefore the introduction
of fluoro-atoms at the 5-, 7-, and 8-positions of polyhalo 2,4-diami-
noquinazolines could generally lead to more potent analogs.
In summary, an efficient, concise, and simple procedure for the
preparation of a highly functionalized 2,4-diaminoquinazoline li-
brary was developed, and the library compounds produced were
proven to have remarkably potent antitumor activity. The substitu-
tion at the 5- and 7-positions with n-BuNH or fluorine atoms, and
at the 8-position with a chlorine atom, on the 2,4-diaminoquinaz-
ole ring, plays an important role in regulating cytotoxic activity.
This makes the compounds 3b and 5a the most promising leads
for future applications, pending further structural modifications
based on the valuable information described here. In addition, 3c
exhibited the most significant anti-HIV-1 activity, which paves
the way to finding promising leads for anti-HIV-1 in the future.
Table 4
Cytotoxic activities of 2,4-diaminoquinazoles 3–5 in vitroa (IC50 g/mL)b
, l
No.
Compound
Skov-3
HL-60
A431
A549
HepG-2
1
2
3
4
5
6
7
8
3a
3b
3c
3d
4a
4b
4c
4d
4e
4f
4g
4h
4i
4j
5a
5b
1.3
0.14
8
0.8
3.7
6.5
1.6
4.8
0.8
11
2.1
2.4
0.4
1.0
0.6
1.6
3.7
2.5
14
19
0.04
51
6.7
47
40
2.2
35
0.3
14
5.2
18
2.3
6.4
0.7
1.4
8.5
7.2
88
62
26
7.7
1.4
3.3
1.5
105
9.5
4.8
7.0
5.1
4.5
1.4
12.9
6.5
11.9
7.3
16.4
2.5
1.0
716
20.7
>1000
68.3
32.4
535
0.6
29.7
30.8
2.5
4.8
5.0
3.7
5.9
1.4
10.2
1.0
1.4
27.8
5.4
16.2
2.0
1.3
1.3
>1000
188
>1000
>1000
103
115
34.1
>1000
313
>1000
7.1
Table 5
9
Anti-HIV-1 replication activity in HIV-1IIIB infected C8166 cell linesa,b
10
11
12
13
14
15
16
17
18
19
20
21
22
23
c
d
No.
Compound
CC50
(lg/mL)
EC50
TIe
1
2
3
4
5
6
7
8
3a
3b
3c
4a
4b
5g
14.3/14.6
3.9/4.0
36.4/105.8
6.3/8.6
1.0/1.8
0.1/0.6
0.6/1.6
0.4/0.6
0.4/0.2
3.2/4.4
13.3/17.3
0.004
8.0/14.0
7.0/32.8
59.6/66.6
13.5/15.7
16.3/31.2
10.6/15.8
11.5/15.1
286250
26.5
2.0
>1000
>1000
13.9
>1000
>1000
>1000
>1000
5.1
2.6
5c
5d
5e
5f
5g
5h
19.2
13.4
>1000
84
5.0
>1000
8.2
7.2/6.9
51.0/46.8
>200/>200
1145
5h
14
AZTf
9.4
1.2
0.5
a
b
c
All data presented are averages of at least three separate experiments.
The data are from Ref. 18.
CC50 (50% cytotoxic concentration), concentration of drug that causes 50%
Cisplatin (DDP)
a
The cytotoxicity, determined as IC50 values for each cell line, is the concentra-
reduction in total C8166 cell number.
d
tion of compound that can reduce the optical density of treated cells by 50% with
respect to untreated cells using the MTT assay.
EC50: concentration that inhibits HIV-1IIIB replication by 50%.
e
In vitro therapeutic index TI = CC50/EC50
.
b
f
Data represent the mean values of three independent measurements.
AZT was used as the reference compound.

S.-J. Yan et al. / Bioorg. Med. Chem. Lett. 20 (2010) 4432–4435
4435
10. Chan, J. H.; Hong, J. S.; Kuyper, L. F.; Baccanari, D. P.; Joyner, S. S.; Tansik, R. L.;
Boytos, C. M.; Rudolph, S. K. J. Med. Chem. 1995, 38, 3608.
Acknowledgments
11. (a) Yan, S.-J.; Huang, C.; Su, C.-X.; Ni, Y.-F.; Lin, J. J. Comb. Chem. 2010, 12, 91; (b)
Yan, S.-J.; Huang, C.; Zheng, X.-H.; Huang, R.; Lin, J. Bioorg. Med. Chem. Lett.
2010, 20, 48; (c) Shu, L.-J.; Mayor, M. Chem. Commun. 2006, 4134; (d) Kitazume,
T.; Nakajima, S. J. Fluor. Chem. 2004, 125, 1447; (e) Chan, P. K.; Leong, W. K.
Organometallics 2008, 27, 1247.
The authors wish to acknowledge financial support in the form
of grants from NSFCs (Nos. 30860342 and 20762013) and NSFYNs
(Nos. 2009cc017 and 2008cd063), and we thank Yong-Tang Zheng
at the Key Laboratory of Animal Models and Human Disease Mech-
anisms of the Chinese Academy of Sciences and Yunnan Province,
Kunming Institute of Zoology, Chinese Academy of Sciences, for
his help regarding the inhibitory activity against HIV-1.
12. Hagmann, W. K. J. Med. Chem. 2008, 51, 4359.
13. General procedure for the synthesis of polyhalo 2,4-diaminoquinazoline 3: A dry
mortar was charged with polyhaloisophthalonitrile
1 (1.5 mmol) and
guanidine carbonate (2.0 mmol). The mixture was mixed at room
temperature by vigorously grinding with a pestle for a few minutes (ca.
5 min). The mixture was placed in a microwave tube and irradiated in a
microwave reactor (Discover), with control of power and temperature by
infrared detection, at 110 °C for 10 min (maximum power 200 W). After
cooling, the reaction mixture was poured into 25 mL of water and filtered to
obtain the crude products, which were purified by column chromatography
(petrol/ethyl acetate = 1:1–1:2, v/v) on silica-gel to give the desired products 3.
Compound 3a: Yellow solid. Mp >300 °C. IR (KBr): 3457, 3392, 3323, 3204,
Supplementary data
Supplementary data associated with this article can be found, in
the online version, at doi:10.1016/j.bmcl.2010.06.056.
2225, 1532, 1562, 1086, 1172 cmÀ1 1H NMR (500 MHz, DMSO-d6) d 6.84 (br,
.
1H, NH2), 7.09 (br, 1H, NH2), 7.41 (br, 1H, NH2), 7.95 (br, 1H, NH2). 13C NMR
(125 MHz, DMSO-d6) d 104.2, 107.8, 115.0, 126.8, 135.2, 135.5, 161.0, 161.7,
162.3. HRMS (TOF ES+): m/z calcd for C9H5Cl3N5 [(M+H+)], 287.9605; found,
287.9608. Compound 3b: Yellow solid. Mp >300 °C. IR (KBr): 3479, 3377, 3160,
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(d) VanBrocklin, H. F.; Lim, J. K.; Coffing, S. L.; Hom, D. L.; Negash, K.; Ono, M. Y.;
Gilmore, J. L.; Bryant, I.; Riese, D. J., II J. Med. Chem. 2005, 48, 7445; (e) Bader, H.;
Freisheimt, J. H. J. Med. Chem. 1991, 34, 1447; (f) Elslager, E. F.; Johnson, J. L.;
Werbel, L. M. J. Med. Chem. 1983, 26, 1753.
2239, 1630, 1561, 1399, 1092, 834 cmÀ1 1H NMR (500 MHz, DMSO-d6) d 7.05
.
(br, 1H, NH2), 7.34 (br, 2H, NH2), 7.41 (br, 1H, NH2), 8.19 (br, 1H, NH2). 13C NMR
(125 MHz, DMSO-d6)
d 82.1, 98.0, 109.7, 110.9, 155.8, 161.1, 159.2 (d,
J = 267.0 Hz), 160.6, 162.2 (d, J = 265.0 Hz), 163.9. HRMS (TOF ESÀ): m/z calcd
for C9H3ClF2N5 [(MÀH+)], 254.0051; found, 254.0050.
14. General procedure for the preparation of products 4: A 25 mL round-bottom flask
was charged with polyhalo 2,4-diaminoquinazoline
3 (1 mmol), N,N-
dimethylformamide (10 mL) and t-BuOK (1.5 mmol), then the solution was
added to nucleophile (1.1 mmol). The mixture was stirred at room temperature
until TLC indicated complete consumption of the starting 3. The mixture was
quenched by the addition of water (20 mL), and then EtOAc (30 mL) was added.
The organic phase was washed with water (10 mL Â 2), dried over Na2SO4,
concentrated and purified by flash column chromatography, to afford the final
product 4 in 61–89% yield. Compound 4a: light yellow solid. Mp 279–283.5 °C.
IR (KBr): 3471, 3384, 3140, 2224, 1662, 1600, 1555, 1504, 1468, 1366, 1242,
842 cmÀ1
.
1H NMR (500 MHz, DMSO-d6) d 6.97–7.26 (m, 8H, Ar, NH2), 7.83 (br,
86.2 (d,
1H, NH2), 8.45 (br, 1H, NH). 13C NMR (125 MHz, DMSO-d6)
d
4. Antonello, A.; Hrelia, P.; Leonardi, A.; Marucci, G.; Rosini, M.; Tarozzi, A.;
Tumiatti, V.; Melchiorre, C. J. Med. Chem. 2005, 48, 28.
J = 18.8 Hz), 96.0, 112.8, 114.4, 118.9, 1122.1, 129.3, 143.0, 143.2, 154.7,
160.2, 162.8 (d, J = 245.0 Hz), 163.2. HRMS (TOF ES+): m/z calcd for C15H11ClFN6
[(M+H+)], 329.0712; found, 329.0717.
5. (a) Liu, F.; Chen, X.; Allali-Hassani, A.; Quinn, A. M.; Wasney, G. A.; Dong, A.-P.;
Barsyte, D.; Kozieradzki, I.; Senisterra, G.; Chau, I.; Siarheyeva, A.; Kireev, D. B.;
Jadhav, A.; Herold, J. M.; Frye, S. V.; Arrowsmith, C. H.; Brown, P. J.; Simeonov,
A.; Vedadi, M.; Jin, J. J. Med. Chem. 2009, 52, 7950; (b) Kung, P.-P.; Casper, M. D.;
Cook, K. L.; Wilson-Lingardo, L.; Risen, L. M.; Vickers, T. A.; Ranken, R.; Blyn, L.
B.; Wyatt, J. R.; Cook, P. D.; Ecker, D. J. J. Med. Chem. 1999, 42, 4705.
6. Kuyper, L. F.; Baccanari, D. P.; Jones, M. L.; Hunter, R. N.; Tansik, R. L.; Joyner, S.
S.; Boytos, C. .; Rudolph, S. K.; Knick, V.; Wilson, H. R.; Caddell, J. M.; Friedman,
H. S.; Comley, J. C. W.; Stables, J. N. J. Med. Chem. 1996, 39, 892.
7. (a) Thurmond, J.; Butchbach, M. E. R.; Palomo, M.; Pease, B.; Rao, M.; Bedell, L.;
Keyvan, M.; Pai, G.; Mishra, R.; Haraldsson, M.; Andresson, T.; Bragason, G.;
Thosteinsdottir, M.; Bjornsson, J. M.; Coovert, D. D.; Burghes, A. H. M.; Gurney,
M. E.; Singh, J. J. Med. Chem. 2008, 51, 449; (b) Okano, M.; Mito, J.; Maruyama,
Y.; Masuda, H.; Niwa, T.; Nakagawa, S.-I.; Nakamura, Y.; Matsuura, A. Bioorg.
Med. Chem. Lett. 2009, 17, 119; (c) Yokoyama, K.; Ishikawa, N.; Igarashi, S.;
Kawano, N.; Masuda, N.; Hattori, K.; Miyazaki, T.; Ogino, S.-I.; Orita, M.;
Matsumoto, Y.; Takeuchi, M.; Ohta, M. Bioorg. Med. Chem. Lett. 2008, 16, 7968;
(d) Häcker, H.-G.; de la Haye, A.; Sterz, K.; Schnakenburg, G.; Wiese, M.;
Gütschow, M. Bioorg. Med. Chem. Lett. 2009, 19, 6102; (e) Smits, R. A.; Adami,
M.; Istyastono, E. P.; Zuiderveld, O. P.; van Dam, C. M. E.; de Kanter, F. J. J.;
Jongejan, A.; Coruzzi, G.; Leurs, R.; de Esch, I. J. P. J. Med. Chem. 2010, 53, 2390.
8. Dener, J. M.; Lease, T. G.; Novack, A. R.; Plunkett, M. J.; Hocker, M. D.; Fantauzzi,
P. P. J. Comb. Chem. 2001, 3, 590.
15. General procedure for the preparation of products 5: A 25 mL round-bottom flask
was charged with polyhalo 2,4-diaminoquinazoline
3 (1 mmol), N,N-
dimethylformamide (10 mL) and t-BuOK (3.0 mmol), then the solution was
added to nucleophile (2.5 mmol). The mixture was stirred at 90 °C until TLC
indicated complete consumption of the starting 3. The mixture was quenched
by the addition of water (20 mL), and then EtOAc (30 mL) was added. The
organic phase was washed with water (10 mL Â 2), dried over Na2SO4,
concentrated and purified by flash column chromatography, to afford the
final product 5 in 73–90% yield. Compound 5a: Light yellow solid. Mp 127–
130 °C. IR (KBr): 3413, 3205, 2959, 2920, 2203, 1594, 1462, 1392, 1113,
796 cmÀ1 1H NMR (500 MHz, DMSO-d6) d 0.74–0.86 (m, 6H, CH3), 1.28–1.30
.
(m, 4H, CH2), 1.50–1.54 (m, 4H, CH2), 2.99–3.00 (m, 2H, NCH2), 3.52–3.54 (m,
2H, NCH2), 4.80 (br, 1H, NH), 5.64 (br, 1H, NH), 6.55 (br, 2H, NH2), 7.74 (br, 2H,
NH2). 13C NMR (125 MHz, DMSO-d6) d 14.0, 19.6, 20.0, 32.3, 32.6, 45.4, 51.1,
86.4, 98.4, 105.9, 118.0, 147.9, 153.3, 153.8, 162.3. HRMS (TOF ES-): m/z calcd
for C17H23ClN7 [(MÀH+)], 360.1709; found, 360.1709.
16. CCDC 760830 contains the supplementary crystallographic data for compound
5b. These data can be obtained free of charge from The Cambridge
Crystallographic Data Center via www.ccdc.cam.ac.uk/data_request/cif.
17. (a) Carmichael, J.; DeGraff, W. G.; Gazdar, A. F.; Minna, J. D.; Mitchell, J. B.
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