Amine substitution of quinazolinones leads to selective nanomolar AChE inhibitors with 'inverted' binding mode

Article abstract of DOI:10.1016/j.bmc.2014.06.045

Selective and nanomolar acetylcholinesterase inhibitors were obtained by connecting tri- and tetracyclic quinazolinones - previously described as moderately active and unselective cholinesterase (ChE) inhibitors - via a hydroxyl group in para position to an anilinic nitrogen with different amines linked via a three carbon atom spacer. These tri- and tetracyclic quinazolinones containing different alicyclic ring sizes and connected to tertiary amines were docked to a high-resolution hAChE crystal structure to investigate the preferred binding mode in relation to results obtained by experimental structure-activity relationships. While the 'classical orientation' locating the heterocycle in the active site was rarely found, an alternative binding mode with the basic aliphatic amine in the active center ('inverted' orientation) was obtained for most compounds. Analyses of extended SARs based on this inverted binding mode are able to explain the compounds' binding affinities at AChE.

Full text of DOI:10.1016/j.bmc.2014.06.045

Bioorganic & Medicinal Chemistry 22 (2014) 4867–4881
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Amine substitution of quinazolinones leads to selective nanomolar
AChE inhibitors with ‘inverted’ binding mode
Fouad H. Darras , Sarah Wehle b, , Guozheng Huang a,b, Christoph A. Sotriffer , Michael Decker
a,
b
a,b,
⇑
a
Institut für Pharmazie, Universität Regensburg, Universitätsstraße 31, D-93053 Regensburg, Germany
Pharmazeutische und Medizinische Chemie, Institut für Pharmazie und Lebensmittelchemie, Julius-Maximilians-Universität Würzburg, Am Hubland, D-97074 Würzburg, Germany
b
a r t i c l e i n f o
a b s t r a c t
Article history:
Selective and nanomolar acetylcholinesterase inhibitors were obtained by connecting tri- and tetracyclic
quinazolinones—previously described as moderately active and unselective cholinesterase (ChE) inhibi-
tors—via a hydroxyl group in para position to an anilinic nitrogen with different amines linked via a three
carbon atom spacer. These tri- and tetracyclic quinazolinones containing different alicyclic ring sizes and
connected to tertiary amines were docked to a high-resolution hAChE crystal structure to investigate the
preferred binding mode in relation to results obtained by experimental structure–activity relationships.
While the ‘classical orientation’ locating the heterocycle in the active site was rarely found, an alternative
binding mode with the basic aliphatic amine in the active center (‘inverted’ orientation) was obtained for
most compounds. Analyses of extended SARs based on this inverted binding mode are able to explain the
compounds’ binding affinities at AChE.
Received 17 April 2014
Revised 18 June 2014
Accepted 22 June 2014
Available online 1 July 2014
Keywords:
Alzheimer’s disease
Cholinesterase inhibitors
Computational study
Heterocycles
Quinazolinones
Ó 2014 Elsevier Ltd. All rights reserved.
1
. Introduction
Alzheimer’s disease (AD) is a progressive, degenerative disorder
of ACh in the brain by inhibiting the neurotransmitter degrading
enzyme acetylcholinesterase (AChE), which is primarily responsi-
ble for its hydrolysis and termination of action. Therefore, acetyl-
cholinesterase inhibitors (AChEIs) reached the pharmaceutical
market and despite their purely symptomatic mode of action still
represent the first-line treatment of AD. Although tacrine was the
first of the cholinesterase inhibitors approved for the treatment
of AD in 1993, its use has been largely abandoned because of a high
incidence of side effects, mainly hepatotoxicity; nevertheless, there
are efforts to counteract tacrine’s liver toxicity using hybrid and
of the brain and the most common form of dementia. With
improvements in health care, longer life expectancy, and continued
rise of the average age of the population, the prevalence of AD is
projected to increase dramatically over the coming years. The glo-
bal prevalence of dementia is estimated to be as high as 24 million,
1
and is predicted to double every 20 years through to 2040. The
main symptoms associated with AD involve a decline in cognitive
function and primarily memory loss. In the later stage of the dis-
ease language deficits, depression, agitation, mood disturbances
and psychosis are often seen. The etiology of AD is not completely
understood; however, there are several diverse pathological factors
7
–10
co-drug molecules.
Three other acetylcholinesterase inhibitors
Ò
(AChEIs) are in clinical use today: donepezil (Aricept ), rivastig-
mine (Exelon ), and galantamine (Razadyne ) are approved for
2
Ò
Ò
1
1
mild to moderate AD. AChEIs prevent the breakdown of ACh
and therefore boost cholinergic neurotransmission in forebrain
regions, which is thought to constitute their clinical benefits.
Numerous novel AChEIs have been described in recent years,12
and especially natural products and alkaloids turned out as a valu-
3
4
such as amyloid-b (Ab) deposits,
tive stress, and decreased levels of acetylcholine (ACh).
s
-protein (
s
) aggregation, oxida-
5
6
According to the cholinergic hypothesis, AD’s pathophysiology
and especially its later stage development of cognitive deficits is
associated with a loss of cholinergic neurotransmission with
decreased levels of acetylcholine in the brain areas dealing with
learning, memory, behavior and emotional responses. Symptom-
atic treatment of AD can be achieved by increasing synaptic levels
1
3–15
able source of novel lead structures.
It has to be emphasized
that the huge majority of novel compounds described—and this
is especially true for natural products and natural lead struc-
tures–are only moderately active and compounds rarely reach
1
2–15
inhibitory activities below the micromolar range.
Thus nano-
molar inhibitors are still desirable and so is information of how
to yield them from lead structures with lower inhibitory activities.
Quinazolines are fused heterocycles that are of considerable
interest in medicinal chemistry because of the diverse range of
⇑
Corresponding author. Tel.: +49 931 31 89676; fax: +49 931 31 85494.
E-mail address: michael.decker@uni-wuerzburg.de (M. Decker).
These authors contributed equally to the publication, F. Darras (synthesis) and
S. Wehle (computational studies).
http://dx.doi.org/10.1016/j.bmc.2014.06.045
0
968-0896/Ó 2014 Elsevier Ltd. All rights reserved.

4
868
F. H. Darras et al. / Bioorg. Med. Chem. 22 (2014) 4867–4881
their biological properties.16,17 The most important derivatives of
quinazoline systems known so far are keto-quinazolines, also
(aliphatic, cyclic , aromatic amines)
(5-, 6- and 7- membered rings)
R1
O
N
1
8,19
called quinazolinones.
The quinazolinone moiety is an impor-
N
O
R2
N
n
tant pharmacophore showing many types of pharmacological
1
8
activities, and the quinazolinones are considered to be ‘privileged
structures’ for drug development.20 The chemistry of 4-oxo
compounds (4(3H)-quinazolinones) has received an increasing
interest because of its biological significance. Derivatives of this
(n= 1, 4)
(tri- and tetracyclic moieties)
Figure 2. Structural modification on the quinazolinone moiety for structure-
2
1
22
21
system show antifungal, antimicrobial, anti-bacterial, anti-
activity relationships.
2
3
24
25
inflammatory, anticonvulsant, and antiproliferative activities.
SARs of the quinazolinone ring system examined in various studies
suggest position 6 suitable for connection to other moieties
this novel class of potent AChE inhibitors and to identify the puta-
tive binding mode at the molecular level for further compound
development.
1
8,26
(Fig. 1).
Our group had synthesized novel tri- and tetracyclic quinazoli-
nones and their corresponding reduced analogs (quinazolines) as
ChEIs derived from the naturally occurring alkaloids deoxyvasici-
SAR studies (Fig. 2) have been performed on two different
classes of heterocyclic compounds, that is, tri- and tetracyclic qui-
nazolinones connected to various tertiary amines by different
spacer lengths. For each moiety, firstly the effect of the nature of
the connected amines was evaluated by connecting the phenolic
quinazolinones with a cyclic amine such as pyrrolidine, piperidine,
and azepane, with diethylamine as aliphatic amine, or 1,2,3,4-
tetrahydroisoquinoline and imidazole as heterocyclic amines,
respectively. Differences in inhibitory activity provide information
on the role of this moiety with regards to molecular interaction
with the enzyme. Secondly, the influence of the ring size of the
quinazolinone’s fused alicyclic ring was evaluated (five-, six-,
seven-membered ring; tri- and tetracyclic fused systems). A puta-
tive correlation of SARs on the heterocycles of this class of
compounds with the ones of unsubstituted quinazolinones and
none 1, dehydroevodiamine chloride 2, and rutaecarpine
3
2
7,28
(Fig. 1).
Deoxyvasicinone 1 (a five-membered ring tricyclic qui-
nazolinone) exhibited a moderate inhibitory activity towards AChE
with an IC50 of 82.5 M, the corresponding six membered-tricyclic
quinazolinone (the alkaloid mackinazolinone) showed an IC50
AChE) of 38.6 M, and the seven membered-tricyclic quinazoli-
none showed an IC50 (AChE) of 279
l
(
l
29
l
M. Low basicity combined
with very poor water solubility and poor to moderate inhibitory
activities made structural modifications of the heterocycle neces-
sary for further development.
Tri- and tetracyclic moieties served as a starting point for stud-
ies to synthesize a novel series of dual acting compounds as AChE
inhibitors/H
shows a high similarity to the aromatic and/or heterocyclic parts
of many known H antagonist ligands. By bridging the phenolic
receptor antagonists.30 The quinazolinone scaffold
3
2
7,29
related heterocycles was investigated.
Thirdly, we also evalu-
3
ated the influence of the spacer length between the phenolic het-
erocycle and the basic amine (three and six carbon atoms spacer
heterocycle via an ether differently sized tertiary amines could
be attached to the heterocycle. This was carried out because the
piperidinylpropoxy phenyl structure represents the key pharmaco-
3
0
length) (Fig. 2).
According to a computational study by AbdulHameed et al.33 on
quinazolinimine-based BChEIs synthesized in our group, the core
heterocycle of quinazolinones, quinazolines and quinazolinimines
binds to the catalytic active site (CAS) at the end of the cholinester-
ase binding gorge. Herein, we assume that the quinazolinones con-
nected to a tertiary amine as phenol ethers (i.e. amino-propoxy
phenyl compounds) might not bind to the CAS in the ‘classical’
binding mode of quinazolinones, but with the tertiary amine being
placed in or near the CAS, which will be referred to as an ‘inverted’
binding mode. An intensive computational study with all synthe-
sized inhibitors was performed to investigate and understand the
binding mode of these sets of compounds, and to provide feedback
to experimentally prove this assumption by synthesizing addi-
tional inhibitors and provide further information for optimization
of this novel class of AChE inhibitors.
phore of a whole class of H
3
antagonists.31 It was found that the
tricyclic basic heterocycle connected to a piperidine via a three car-
bon atom spacer (compound 4; Fig. 1) was the most potent com-
pound at both targets with K
3
i
(hH
3.9 nM, and IC50 (eeAChE) = 66.5 nM, respectively.
In accordance with SARs on H
antagonists32 reduced basicity of
affinity. This was indeed also the case
affinities of quinazolinones are
3
) = 76.2 nM, IC50 (hAChE) =
30
3
the heterocycle decreases H
3
for our quinazolinones, where H
3
around 40 fold lower than the affinity of their corresponding
totally reduced forms (reduction of the carbonyl group and the
3
0
imine double bond).
The dual-acting compounds also exhibit considerable inhibitory
activities at butyrylcholinesterase. For the quinazolinones con-
nected to tertiary amines and presented herein, a good to excellent
selectivity over BChE was observed (cf. Table 2). Therefore, the
peculiar selectivity both over H
3
as well as BChE combined with
2. Chemistry
the high affinity at AChE prompted us to investigate the SARs of
A straightforward synthesis for 6-substituted quinazolinones
was developed using 6-(benzyloxy)-1H-benzo[d][1,3]oxazine-2,4-
dione 5 (Scheme 1) as starting material. It was synthesized in four
O
5
O
O
steps starting from 2-amino-5-hydroxybenzoic acid with overall
4
a
3
N
6
7
N
N
N
N
30,34
4
almost quantitative yield as previously described.
The result-
ing isatoic anhydride 5 represents an activated form of anthranilic
acid with a benzyl protected phenolic OH. The benzylation of the
phenolic group in compound 5 is very advantageous for subse-
quent synthetic steps by improvement of solubility of the resulting
isatoic anhydride.
Tetracyclic compounds (Scheme 1A) were synthesized by sol-
ventless fusion of isoindolin-1-one, 3,4-dihydroisoquinolin-1(2H)-
one or 2,3,4,5-tetrahydro-1H-benzo[c]azepin-1-one, respectively,
with compound 5 under microwave conditions with 200 W irradi-
ation power at 220 °C for one hour to get the benzylated tetracyclic
8
a N
2
N
H
N
H
8
1
H C
3
1
2
Cl
3
N
O
N
N
4
Figure 1. Chemical structures of deoxyvasicinone 1, dehydroevodiamine chloride 2,
rutaecarpine 3 and dual-acting AChEI and hH antagonist 4.
3

F. H. Darras et al. / Bioorg. Med. Chem. 22 (2014) 4867–4881
4869
AO
B
O
n
O
O
N
n
O
HN
O
N
O
N
n
HO
O
N
O
O
(ii)
HO
O
N
N
N
HN
O
O
n
N
H
O
(ii)
N
N
H
5
O
O
n
n
(i)
5
(i)
33, n = 0
34, n = 1
35, n = 2
6
7
8
, n = 0
, n = 1
, n = 2
36, n = 0
37, n = 1
38, n = 2
9
, n = 0
(iii)
1
0, n = 1
(iii)
1
1, n = 2
O
Br
O
1
5, n = 0, R = piperidine
6, n = 0, R = azepane
7, n = 0, R = pyrrolidine
8, n = 0, R = diethylamine
42, n = 0, R = piperidine
43, n = 0, R = azepane
44, n = 0, R = pyrrolidine
N
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
O
N
n
N
Br
O
39, n = 0
40, n = 1
1, n = 2
n
N
45, n = 0, R = diethylamine
4
9, n = 0, R = 1,2,3,4-tetrahydroisoquinoline
0, n = 0, R = imidazole
46, n = 0, R = 1,2,3,4-tetrahydroisoquinoline
47, n = 0, R = imidazole
1
1
1
2, n = 0
3, n = 1
4, n = 2
1, n = 1, R = piperidine
48, n = 1, R = piperidine
(iv)
2, n = 1, R = azepane
49, n = 1, R = azepane
3, n = 1, R = pyrrolidine
50, n = 1, R = pyrrolidine
4, n = 1, R = diethylamine
5, n = 1, R = 1,2,3,4-tetrahydroisoquinoline
6, n = 1, R = imidazole
(iv)
n
51, n = 1, R = diethylamine
52, n = 1, R = 1,2,3,4-tetrahydroisoquinoline
53, n = 1, R = imidazole
O
N
R
O
7, n = 2, R = piperidine
54, n = 2, R = piperidine
N
O
N
8, n = 2, R = azepane
55, n = 2, R = azepane
9, n = 2, R = pyrrolidine
R
O
56, n = 2, R = pyrrolidine
n
N
0, n = 2, R = diethylamine
1, n = 2, R = 1,2,3,4-tetrahydroisoquinoline
2, n = 2, R = imidazole
57, n = 2, R = diethylamine
58, n = 2, R = 1,2,3,4-tetrahydroisoquinoline
59, n = 2, R = imidazole
Scheme 1. (A) Synthesis of tetracyclic quinazolinones, reagents and conditions: (i) MW, 200 W, 200 °C, 1 h; (ii) H
2
, Pd/C, ethanol, rt, 24 h; (iii) 1,3-dibromopropane, K
2
CO
3
, KI,
acetonitrile, 90 °C, 3 h; (iv) amine, K CO , KI, acetonitrile, 90 °C, 3 h. (B) Synthesis of tricyclic quinazolinones, reagents and conditions: (i) MW, 200 W, 200 °C, 1 h; (ii) H
2
3
2
, Pd/C,
ethanol, rt, 24 h; (iii) 1,3-dibromopropane, K
2
3 2 3
CO , KI, acetonitrile, 90 °C, 3 h; (iv) amine, K CO , KI, acetonitrile, 90 °C, 3 h.
Table 1
3
. AChE and BChE inhibition
hAChE inhibition results of selected tri- and tetracyclic compounds
Compd hAChE inhibition [IC50
pIC50 ± SEM)]
Compd hAChE inhibition [IC50
(pIC50 ± SEM)]
All target compounds were tested for their ability to inhibit ace-
tylcholinesterase (eeAChE, EC.3.1.1.7 from electric eel) and butyr-
ylcholinesterase (eqBChE, EC.3.1.1.8 from equine serum) (Table 2)
(cf. SI for sequence alignment).
Additionally, selected compounds (quinazolinones with a pipe-
ridinyl, diethylamino or one imidazole substituent, respectively)
were additionally tested for inhibition of human acetylcholinester-
ase (hAChE) to confirm binding profiles directly at the human
enzyme (Table 1). It was found that tacrine shows a threefold
lower potency (IC50 = 71.5 nM) on the human enzyme compared
to the IC50 value of 24.1 nM measured on eeAChE. The piperidinyl
inhibitors 15, 21, 27, 42, 48 and 54 showed 7 to 39 fold higher
inhibitory activities toward the electric eel derived AChE (Tables
(
Tacrine 71.5 nM (7.145 ± 0.038)
42
45
48
51
54
57
61
1.1
1.2
l
l
M (5.966 ± 0.183)
M (5.905 ± 0.136)
15
18
21
24
27
30
32
142 nM (6.845 ± 0.144)
200 nM (6.697 ± 0.11)
216 nM (6.664 ± 0.085)
669 nM (6.174 ± 0.149)
535 nM (6.272 ± 0.087)
225 nM (6.648 ± 0.211)
1.5 M (5.815 ± 0.253)
820 nM (6.086 ± 0.219)
12.1 M (4.918 ± 0.045)
35% inhibition at 100 lM
l
l
4.2
l
M (5.379 ± 0.338)
M (4.248 ± 0.167)
56.5
l
compounds 6, 7 and 8, respectively, with different fused ring sizes.
Quantitative debenzylation by catalytic hydrogenation over Pd/C in
ethanol at room temperature gave the corresponding phenolic tet-
racyclic compounds 9, 10 and 11. Williamson ether synthesis by
reaction of compounds 9, 10 and 11 with 1,3-dibromopropane in
presence of potassium carbonate as a base with a catalytic amount
of potassium iodide in acetonitrile as solvent under reflux for three
hours gave compounds 12, 13, and 14, respectively. The last syn-
thetic step was nucleophilic substitution by reaction of bromo
compounds 12, 13, 14 with different secondary amines using the
above alkylation conditions. The tetracyclic five-membered ring
target compounds 15–20 were obtained by reaction of compound
1
and 2). This difference in potency was not seen before for related
nitrogen-bridgehead compounds and is not immediately plausible
from the structural point of view due to highly conserved amino
3
0
acids in the binding site of these two species. A difference in inhi-
bition data on these two AChEs from different species was previ-
ously also described for pyridonepezils and quinolinodonepezils
3
5,36
and a set of tacrine dimers.
4. Computational binding mode analysis
1
2 with secondary amines (piperidine, azepane, pyrrolidine, dieth-
ylamine, 1,2,3,4-tetrahydroisoquinoline and imidazole). The tetra-
cyclic six-membered ring target compounds 21–26 were obtained
by reaction of compound 13 with the respective secondary amines,
and tetracyclic seven-membered ring target compounds 27–32 by
reaction of compound 14 (Scheme 1A).
Similarly, tricyclic compounds (Scheme 1B) were synthesized
by solventless fusion of pyrrolidin-2-one, piperidin-2-one or aze-
pan-2-one with compound 5 under microwave conditions to yield
the benzylated tricyclic compounds 33, 34 and 35, respectively.
Quantitative debenzylation by catalytic hydrogenation gave the
corresponding phenolic tricyclic compounds 36, 37, 38. William-
son ether synthesis under the same conditions described above
gave compounds 39, 40, 41, respectively. Tricyclic target com-
pounds 42–59 were synthesized by nucleophilic substitution reac-
tion of the bromo spacer compounds 39, 40 and 41, respectively,
with the corresponding secondary amines using the same condi-
tions described above for the tetracyclic target compounds
Docking studies were carried out to investigate whether the
compounds of the inhibitor library show a common binding mode
preference with respect to AChE. In principle, two different modes
of inhibition are imaginable: first, a ‘classical’ binding mode
corresponding to the placement of the tri- or tetracyclic scaffold
near the catalytic active site (CAS), assumed for tri- and tetracyclic
ChE inhibitors without an alkylamine, for example, unsubstituted
quinazolinones (cf. Fig. 4); and, second, an inverted binding
mode where instead of the aromatic scaffold the protonated ter-
tiary amine is positioned at the CAS deep in the binding gorge.
Interestingly, as further detailed below, docking shows a clear
preference for the inverted binding mode for the vast majority of
the library compounds: among the top-five docking poses obtained
for each compound, inverted binding was observed in 77%
(tricyclic compounds) and 99% (tetracyclic compounds) of the
cases, respectively.
As shown in Figure 3 for inhibitor 15 as an example, the
inverted binding mode is characterized by placement of the alicy-
(Scheme 1B).

Table 2
eeAChE and eqBChE inhibition results and the resulting ChE selectivity of tetra- and tricyclic target compounds
O
O
N
n
n
R
O
R
O
N
N
N
Compd
Tacrine
n
R
eeAChEa [IC50 (pIC50 ± SEM)]
eqBChEa [IC50 (pIC50 ± SEM)]
Selectivityb
0.2
Compd
n
R
eeAChEa [IC50 (pIC50 ± SEM)]
eqBChEa [IC50 (pIC50 ± SEM)]
Selectivityb
24.1 nM (7.619 ± 0.081)
3.7 nM (8.436 ± 0.012)
1
5
6
0
0
N
6.9 nM (8.161 ± 0.03)
5.0 nM (8.305 ± 0.065)
4.14
1.87
l
M (3.383 ± 0.028)
M (5.727 ± 0.069)
600
374
42
43
0
0
N
69.2 nM (7.160 ± 0.152)
50.2 nM (7.299 ± 0.072)
122
l
M (3.912 ± 0.145)
1767
104
1
N
l
N
5.22
l
M (5.282 ± 0.024)
M (4.977 ± 0.039)
1
7
8
0
0
N
16.2 nM (7.790 ± 0.027)
11.0 nM (7.961 ± 0.036)
2.40
1.88
l
M (5.620 ± 0.007)
M (5.726 ± 0.145)
148
171
44
45
0
0
N
50.0 nM (7.301 ± 0.073)
101 nM (6.997 ± 0.059)
10.55
l
211
8
1
N
l
N
829 nM (6.082 ± 0.066)
776 nM (6.110 ± 0.053)
N
N
N
N
N
N
1
9
0
227 nM (6.644 ± 0.047)
1.73
9.11
l
M (5.763 ± 0.043)
M (5.041 ± 0.06)
8
46
0
868 nM (6.062 ± 0.066)
1
2
0
1
0
1
N
454 nM (6.343 ± 0.013)
5.5 nM (8.263 ± 0.065)
l
20
47
48
0
1
N
1.90
lM (5.721 ± 0.017)
14.65
lM (4.834 ± 0.040)
8
N
N
N
N
N
N
2
N
955 nM (6.020 ± 0.051)
584 nM (6.233 ± 0.021)
955 nM (2.380 ± 0.008)
174
N
33.3 nM (7.478 ± 0.052)
58.5 nM (7.233 ± 0.033)
6.55
1.77
l
M (5.184 ± 0.028)
M (5.573 ± 0.021)
197
2
2
1
N
13.0 nM (7.886 ± 0.128)
45
49
1
N
l
30
2
3
4
1
1
N
N
91.1 nM (7.041 ± 0.026)
47.5 nM (7.323 ± 0.052)
11
46
50
51
1
1
N
N
210 nM (6.679 ± 0.059)
43.0 nM (7.367 ± 0.038)
16.08
l
M (4.794 ± 0.013)
77
2
2.16
1.83
l
M (5.666 ± 0.047)
M (5.737 ± 0.051)
9.14
5.49
l
M (5.039 ± 0.015)
M (5.261 ± 0.024)
213
2
5
1
625 nM (6.204 ± 0.038)
l
3
52
1
723 nM (6.141 ± 0.035)
l
8
2
6
7
1
2
N
2.44
lM (5.613 ± 0.103)
8.68
2.67
l
M (5.061 ± 0.092)
M (5.573 ± 0.027)
4
53
54
1
2
N
4.24
lM (5.373 ± 0.013)
50.05
12.73
l
M (4.229 ± 0.035)
M (4.895 ± 0.032)
12
2
N
57.1 nM (7.244 ± 0.021)
53.4 nM (7.272 ± 0.042)
l
47
N
43.0 nM (7.366 ± 0.055)
39.2 nM (7.407 ± 0.030)
l
296
2
8
2
N
865 nM (6.063 ± 0.017)
16
55
2
N
1.80
l
M (5.744 ± 0.048)
46
2
9
0
2
2
N
N
529 nM (6.277 ± 0.054)
72.1 nM (7.142 ± 0.046)
3.61
5.10
l
M (5.443 ± 0.021)
M (5.293 ± 0.024)
7
56
57
2
2
N
N
295 nM (6.531 ± 0.050)
50.6 nM (7.296 ± 0.069)
12.19
10.75
l
M (4.914 ± 0.067)
M (4.969 ± 0.059)
41
3
l
71
l
213
3
3
6
1
2
1
2
2
0
1.19
lM (5.925 ± 0.021)
1.84
lM (5.735 ± 0.032)
2
58
2
2
0
653 nM (6.185 ± 0.029)
3.03
l
M (5.519 ± 0.067)
5
N
10.27
lM (4.989 ± 0.018)
52.01
l
M (4.284 ± 0.045)
5
59
N
3.95
0.39
l
M (5.403 ± 0.048)
M30 (6.41 ± 0.05)
59.75
lM (4.224 ± 0.086)
15
22
6030
N
l
8.6 l
M30 (5.07 ± 0.01)
34% inhibition at. 50
l
M
No inhibition at. 50
lM
—
6
a
AChE from electric eel and BChE from equine serum; data are the means of at least three independent determinations.
Selectivity ratio [IC50 (BChE)/IC50 (AChE)].
b

F. H. Darras et al. / Bioorg. Med. Chem. 22 (2014) 4867–4881
4871
clic amine at the CAS near Ser203 and His447 (hAChE numbering)
and within cation– -interaction distance to Trp86, a residue
known for attracting the quaternary amine of the natural substrate
p
3
7
acetylcholine. The tetracyclic scaffold is sandwiched between
Trp286 and Tyr341, showing ring-ring distances suitable for
p–p
interactions with Trp286, while the carboxamide carbonyl oxygen
is stabilized by interactions with conserved water molecules.
Analysis of the experimental SARs support this binding mode, as
discussed in detail below.
5
. Results and discussion
During our intensive SAR studies on the heterocyclic moiety we
had been surprised to observe that connecting the tricyclic quinaz-
olinone moiety with different cyclic amines by three carbon atom
spacers improved AChE inhibition dramatically compared to the
unsubstituted quinazolinones (Fig. 4).30
Different factors such as spacer length, the nature of the alicy-
Figure 3. Inhibitor 15 is illustrated as an example of the smallest tetracyclic
clic amine, calculated pK
a
values, synthesis of a carbocyclic analog
scaffold with the typical interactions found in the inverted binding mode: a 4.0 Å
cation–
carbonyl with water molecule 954, an additional 3.7 Å contact to water molecule
52, and a 3.5 Å -interaction with Trp286. Distances are specified in Å as italic
p interaction with Trp86, a 2.6 Å hydrogen bond of the carboxamide
of 15, and docking studies helped to understand the characteristics
of the amine-substituted quinazolinone class of compounds as
AChE inhibitors and their putative binding mode.
9
p–p
38
numbers. All figures showing docking poses were created with Pymol.
The spacer length of three carbon atoms for the compounds of
the inhibitor library was chosen on the basis of results obtained
for heterocycles with dual-acting H
3
antagonist/AChE inhibiting
properties. To support the suitability of this choice, compound
0 was prepared as an analog of 42 with a six- instead of a
3
0
6
O
O
N
three-carbon-atom spacer between the quinazolinone and the
basic center. Compound 60 was found to be less active, with an
IC50 (eeAChE) of 390 nM compared to 69.2 nM for 42 (Table 2).
The target compounds obtained by connection of tri- and
tetracyclic quinazolinone moieties with different cyclic amines
N
O
N
N
N
n
n
n = 1, IC50 (AChE) = 82.5 µM
n = 2, IC50 (AChE) = 38.6 µM
n = 3, IC50 (AChE) = 279 µM
42, n = 1, IC50 (AChE) = 69.2 nM
48, n = 2, IC50 (AChE) = 33.3 nM
54, n = 3, IC50 (AChE) = 43.0 nM
(
pyrrolidine, piperidine, azepane) as basic center showed inhibi-
tion activity between one and three digit nanomolar IC50 values
at eeAChE (from IC50 = 5.0 nM for compound 16 to 529 nM for com-
pound 29, cf. Table 2). Connection of the corresponding quinazoli-
nones with diethylamine as basic center led to a similar inhibition
range. In contrast, all target compounds from both tri- and
tetracyclic quinazolinones with different fused ring sizes
connected to 1,2,3,4-tetrahydroisoquinoline or imidazole showed
only submicromolar to ten micromolar inhibition at AChE (from
IC50 = 0.65
Figure 4. Acetylcholinesterase inhibitory activities of quinazolinones and their
corresponding piperidinylpropoxy derivatives.
29,30
mined activity. Calculations with MoKa39 provide the following
pK values for the amine substituents: N,N-diethylamine 9.84, pyr-
a
rolidine 9.84, piperidine 9.71, azepane 9.58, 1,2,3,4-tetrahydroiso-
quinoline 8.68, and imidazole 7.07. These values correlate well
with the pIC50 trends observed in homologous compound series
(e.g., cf. the pIC50 values of the tricyclic compounds 42–47 with
fused 6-membered ring: 7.00–7.30 with N,N-diethylamine, pyrrol-
idine, piperidine, or azepane as substituent; 6.06 with tetrahydro-
isoquinoline; and 5.72 with imidazole). Obviously, the influence of
the basicity is modulated by steric factors determining the position
of the charged amine with respect to the aromatic side chain to
lM for compound 58 to 10.27
lM for compound 32) (Table 2).
Interestingly, compounds with piperidine as amine substituent
display the highest potency for AChE and, in particular, the best
selectivity over BChE. The seven-membered ring compounds show
activity in the same range, while the pyrrolidino compounds (with
exception of compound 44) have lower potencies, irrespective of
the connected heterocycle. The fact that the nature of the con-
nected tertiary amine has great influence on the inhibitory activity
of the molecule (one to two digit nanomolar in case of aliphatic
amine; submicromolar to two digit micromolar in case of imidaz-
ole or tetrahydroisoquinoline) is likely to be due to a binding mode
in which the amine is placed in the CAS region.
form cation–p interactions. While the small amines (pyrrolidine,
piperidine, azepane, diethylamine) can all place their nitrogen at
well-suited distances with respect to Trp86, steric hindrance pre-
vents an optimal position in case of the 1,2,3,4-tetrahydroisoquin-
oline substituent (19, 25, 31, 46, 52, 58), leading to less favorable
interactions and weaker activity than expected on the basis of
the amine basicity.
To provide further experimental evidence for the role of the
basic center in the inverted binding mode, compound 61 was
synthesized as carbon analog of compound 15 by replacing
the piperidine by a cyclohexyl ring. Impressively, 61 loses most
of the classes’ inhibitory potency, showing only 34% inhibition
Docking studies indeed point to an inverted binding mode as
the clearly preferred orientation for the vast majority of the
compounds. The notion of such an inverted binding mode is sup-
ported by various observations with respect to the experimental
SARs: First of all, a positively charged tertiary amine is important
for high-affinity binding. Compounds with a neutral imidazole
39
substituent (pK
weaker affinity. This can be interpreted in terms of an essential cat-
ion- -interaction, most likely with Trp86 which is known to play a
similar role in other AChE-complexes.
a
= 7.07 based on a MoKa calculation ) show much
of eeAChE at 50 lM, while compound 15 displays an IC50
p
(eeAChE) of 6.9 nM (Fig. 5). This dramatic activity difference
strongly supports the essential role of the basic nitrogen, which
can most plausibly be rationalized by a position at the CAS of
AChE.
37,40
The importance of amine basicity for potent AChE inhibition is
reflected by a qualitative correlation with experimentally deter-

4
872
F. H. Darras et al. / Bioorg. Med. Chem. 22 (2014) 4867–4881
O
N
O
N
O
O
N
N
N
6
1
1
5
IC50 (AChE) = 6.9 nM
IC50 (BChE) = 4.14 µM
IC50 (AChE) = 34 % inhibition at 50 µM
IC50 (BChE) = no inhibition at 50 µM
Figure 5. Effect of presence and absence of the basic center connected to the
quinazolinone scaffold on AChE inhibitory activity.
Docking of compound 61 showed that, in principle, a binding
mode as obtained for 15 may also be assumed for 61 (rms
deviation: 0.90 Å), with the cyclohexyl ring located in the CAS
region near Trp86. However, in comparison to 15 (score 103.06)
compound 61 is scored much less favourably by GoldScore (score
9
2.20), pointing to the lack of the cation–p interaction and a
considerably weaker affinity. While the other scoring functions
Figure 7. Inhibitor 42 is illustrated as an example of a compound with the smallest
tricyclic scaffold and the inverted binding mode. The most important interactions
available in GOLD besides GoldScore (ChemPLP, ASP, Chem-
4
1–46
Score)
were not able to reflect the affinity difference between
consist of a 4.1 Å cation–
p distance in the CAS region, a 2.8 Å distance of the
the two compounds, the GoldScore values obtained for the docked
carboxamide carbonyl oxygen to water molecule 954 and a hydrogen bridge of
poses of 15 and the other piperidinyl substituted compounds (21,
3.3 Å to water molecule 952. Distances are specified in Å as italic numbers.
2
7, 42, 48, 54) in the inverted binding mode correlate very well
2
(
R = 0.92) with the pIC50, indicating that the modulation of the
oxygen) and water molecule 954. This might contribute to their
weaker inhibition potency compared to the other compounds of
the same scaffold group.
affinity by the different scaffolds is well captured by this scoring
function and the binding mode obtained from docking (cf. SI Fig. 1).
For the tetracyclic scaffolds (15–32) a very clear preference for
the inverted binding mode is found by docking. Within the top ten
scored poses of each of these 18 inhibitors, the majority shows the
The hydrophobic interaction with Trp286 seen for most of the
tetracyclic compounds (with exception of 18, 20, 25, 28 and 31)
is in accordance with published distance preferences (3.4–
4
7
inverted binding mode with cation–
.3–4.6 Å between the protonated alicyclic amine and Trp86, sta-
bilization of the quinazolinone scaffold by water molecule 954
cf. the Experimental part and SI Fig. 2 for details regarding the
structural water molecules) and—in most of the cases—putative
interactions (3.2–3.5 Å) between the fused benzene moieties
p
interaction distances of
3.6 Å), supporting the assumption of a p–p interaction. With
3
the sterically more demanding quinazolinones (with fused six-
and seven-ring) this interaction seems to be particularly favored
and shows best geometries in cases where the scaffold nitrogen
instead of the carbonyl group is pointing to the water molecules
(Fig. 6).
(
p–p
of the inhibitors and the indole side chain of Trp286 located at
the entrance of the binding site. In more detail, inhibitors 15–20
with the smallest tetracyclic scaffold (corresponding to the most
potent compounds of the library) show the carboxamide carbonyl
pointing toward the water molecules, thus forming a 2.4 Å (18),
Also for the inhibitors with the tricyclic scaffolds 42–59 the
inverted binding mode is preferred over the classical one (Fig. 7).
Due to the smaller scaffold size compared to the one of the tetra-
cycles, however, slightly more poses with the classical binding
mode are found here. For the top-ten poses of the inhibitors bear-
ing the azepane, pyrrolidine, or tetrahydroisoquinoline ring, the
inverted binding mode is found exclusively. For the other inhibi-
tors (42, 45, 47, 48, 51, 53, 54, 57, 58 and 59) between one and nine
poses with the classical binding mode are found among the ten
top-scoring poses.
2
.6 Å (15, 16) and weak 3.5 Å (19, with an additional 2.9 Å interac-
tion to water 952) hydrogen bond to water molecule 954 (cf.
Fig. 3). Inhibitors 17 and 20 show a turned scaffold orientation in
the binding pocket, forming a 2.9 Å and 2.8 Å interaction, respec-
tively, between the scaffold nitrogen (instead of the carbonyl
Figure 6. Due to the improved
p–p interactions of the fused benzene and Trp286, the carboxamide carbonyl of the inhibitors 21 (a) and 27 (b) does not point to the water
molecules, but to the opposite direction. Distances are specified in Å as italic numbers.

F. H. Darras et al. / Bioorg. Med. Chem. 22 (2014) 4867–4881
4873
The chosen poses with inverted binding mode show a 2.5–3.1 Å
distance of the carboxamide carbonyl oxygen to water molecule
showing the inverted binding mode similar to the five- and six-
membered ring analogs 42–53.
9
54 and a 3.3–4.5 Å distance to water molecule 952, as shown
As described in the Discussion, from the four scoring functions
4
5,46
exemplarily for ligand 42 in Figure 7. For the piperidine-,
azepane- and pyrrolidine-rings the distance to Trp86 of the choline
binding site varies only between 3.9 and 4.2 Å, whereas for the
N,N-diethylamine substituent the observed distances range from
provided by GOLD, the GoldScore function
was chosen for the
docking studies because it performed best with respect to relative
affinity ranking. In all redocking experiments using GoldScore the
top-ranked docking pose was deviating less than 0.91 Å from the
corresponding crystal structure. All docking poses were clustered
with a 1.5 Å cluster-cut-off by applying the complete linkage
method.
3
.9 to 4.7 Å. Due to the bulkiness of the substituent, a close contact
between the basic nitrogen and Trp86 is prevented for the tetrahy-
droisoquinoline-bearing inhibitors, possibly also explaining their
comparatively weak inhibition potency (IC50(eeAChE) from 653 to
In some cases not the absolute top pose of the inverted binding
mode was chosen, but a pose displaying better hydrogen bond dis-
tances (2.6–3.0 Å) in agreement with experimentally observed
8
68 nM).
4
7
6
. Conclusion
preferences. These poses vary on average by 0.91 score units
and 1.33 Å with respect to the top pose, i.e. they belong to the same
cluster. Distance measurements were carried out in Pymol.38 For
The inhibitory activity and selectivity profile at AChE of previ-
ously described moderately active and unselective tri- and tetracy-
clic quinazolinones were dramatically improved by connecting
them with different amines linked by three carbon atoms. Based
on the experimental findings and the docking studies we assume
that the inverted binding mode is preferred in terms of protein–
aromatic rings putatively involved in p–p or cation–p interactions
the geometric centers were used for distance measurements.
7.2. Enzyme inhibition (acetyl- and butyrylcholinesterase
inhibition assay)
ligand interactions. Besides cation–p and p–p interactions with
Trp residues, structural water molecules can form hydrogen bonds
with the ligand, especially with the carboxamide carbonyl oxygen,
to mediate interactions with the AChE binding site.
AChE (E.C.3.1.1.7, Type VI-S, from Electric Eel) and BChE
(E.C.3.1.1.8, from equine serum) were purchased from Sigma-
Aldrich (Steinheim, Germany). DTNB (Ellman’s reagent), ATC and
BTC iodides were obtained from Fluka (Buchs, Switzerland). The
assay was performed as described in the following procedure:2
Based on the affinity loss for the cyclohexyl-substituted
inhibitor and the much weaker affinity for the (neutral) imidazole
derivative compared to the corresponding piperidinyl analog, a
protonated nitrogen is found to be essential for high affinity
binding at the choline binding site. For the development of novel
AChE inhibitors with a basic center it seems therefore crucial to
investigate the putative binding mode by molecular docking and
by SARs in order to be able to apply the most promising structural
modifications to the inhibitors for further compound development.
7,29
Stock solutions of the test compounds were prepared in ethanol,
À3
100
l
L of which gave a final concentration of 10 M when diluted
to the final volume of 3.32 mL. The highest concentration of the
À4
test compounds applied in the assay was 10 M (10% EtOH in
the stock solution did not influence enzyme activity). In order to
obtain an inhibition curve, at least five different concentrations
À4
À9
(normally 10 –10 M) of the test compound were measured at
5 °C and 412 nm, each concentration in triplicate. For buffer
2
7
7
. Experimental part
preparation, 1.36 g of potassium dihydrogen phosphate (10 mmol)
were dissolved in 100 mL of water and adjusted with NaOH
.1. Computational docking
to pH = 8.0 ± 0.1. Enzyme solutions were prepared to give
À1
2
.5 units mL in 1.4 mL aliquots. Furthermore, 0.01 M DTNB solu-
4
8
Docking studies were performed with GOLD v5.1 and v5.2 fol-
tion, 0.075 M ATC and BTC solutions, respectively, were used. A
cuvette containing 3.0 mL of phosphate buffer, 100 L of the
respective enzyme, and 100 L of the test compound solution
was allowed to stand for 4.5 min, then 100 L of DTNB were added,
and the reaction was started by addition of 20 L of the substrate
lowing the previously described setup.30 50 docking poses were
generated for each compound of the inhibitor library using the
crystal structure 4EY7 of human AChE and seven conserved water
molecules, as described in detail in a preceding study (cf. SI Fig. 1
for illustration of the hydrogen bond network formed by the water
molecules in the AChE binding site). The protonation of the ligands
l
l
l
l
solution (ATC/BTC). The solution was mixed immediately, and
exactly 2.5 min after substrate addition the absorption was mea-
(
15–32 and 42–59) was set according to the expected ionization
sured. For the reference value, 100
compound solution. For determining the blank value, additionally
100 L of water replaced the enzyme solution. The inhibition curve
was obtained by plotting the percentage enzyme activity (100% for
the reference) versus logarithm of test compound concentration.
The same testing protocol was used for hAChE purchased from
Sigma–Aldrich (Steinheim, Germany).
lL of water replaced the test
state at pH 8 (corresponding to the pH of the inhibition assay). This
implied protonation at the tertiary amine for all compounds, con-
sistent with their pK
with MoKa. Based on a titration curve calculation, also performed
with the program MoKa, the imidazole moiety is protonated to
1
l
a
values between 8.68 and 9.84, as calculated
4
9
0.6% and neutral to 89.4% at pH = 8. For the binding mode discus-
sion only the predominant neutral form of the imidazole ring is
considered. For the tricyclic inhibitors 54–59, bearing a fused
seven-membered alicycle, a conformational search was performed
7.3. Chemistry
4
9
in MOE, which resulted in six different conformational families
cf. SI Fig. 3). The MMFF94s force field was used and the conform-
General methods
(
Common reagents and solvents were obtained from commercial
suppliers and used without further purification. Reactions were
conducted using dried flasks under a positive atmosphere of nitro-
gen. Reaction progress was monitored by analytical thin layer
chromatography (TLC) on precoated silica gel GF254 plates. For
detection iodine vapor, or UV light (254 nm), were used. Melting
points are uncorrected and were measured in open capillary tubes,
using a Barnstead Electrothermal IA9100 melting point apparatus.
ers were generated in a stochastic search with 10,000 iterations
and optimized to an rms gradient of 0.01 kcal/(mol Å). Each con-
former was additionally locally minimized with the MMFF94x
force field to an rms gradient of 0.001 kcal/(mol Å) for consistency
with the general ligand setup.30 All six conformers were docked
with all of the tertiary amine moieties to the 4EY7 binding site.
The conformer with a twist-like orientation of the seven mem-
bered ring proved to give consistent docking results of the ligand
1
13
H and C NMR spectral data were obtained from a Bruker

4
874
F. H. Darras et al. / Bioorg. Med. Chem. 22 (2014) 4867–4881
Advance spectrometer (300 MHz and 75 MHz, respectively). Chem-
ical shifts are expressed in ppm relative to CDCl or DMSO-d
7.26:2.50 and 77.16:39.52 ppm for H and C NMR, respectively).
7.3.5. 7-(Benzyloxy)-2,3-dihydropyrrolo[2,1-b]quinazolin-
9(1H)-one (33)
3
6
1
13
(
Starting from pyrrolidin-2-one (1.00 g, 12.25 mmol) and 6-ben-
zyloxyisatoic anhydride 5 (1.00 g, 3.71 mmol). The title compound
was obtained as a yellow solid (450 mg, 78% yield). Mp 188–
ESI-MS samples were analyzed using electrospray ionisation ion-
trap mass spectrometry in nanospray mode using a Thermo Finni-
gan LCQ Deca. The CHN analyses were undertaken using Perkin
Elmer Elemental Analyser PE2400CHNS. In the elemental analysis,
small amounts of solvent were taken into account which had been
applied in previous column chromatography or for transferring the
compounds to another vessel. For column chromatography, silica
gel 60, 230–400 mesh (Merck) was used.
+
1
3
190 °C. ESI-MS: 293.0 m/z [MH] . H NMR (300 MHz, CDCl ) d:
7.75 (d, J = 2.9 Hz, 1H), 7.65–7.58 (m, 1H,), 7.47 (dd, J = 8.2,
1.2 Hz, 1H), 7.44–7.29 (m, 5H), 5.17 (s, 2H), 4.30–4.11 (m, 2H),
1
3
3.25–3.08 (m, 2H), 2.39–2.20 (m, 2H) ppm. C NMR (75 MHz,
CDCl ) d: 160.76, 157.41, 157.18, 149.10136.34, 131.25, 128.66,
3
128.24, 128.20, 127.71, 124.90, 107.14, 70.51, 46.57, 32.25,
19.69 ppm.
6
-(Benzyloxy)-1H-benzo[d][1,3]oxazine-2,4-dione 5 was syn-
thesized in four steps with almost quantitative overall yield, start-
ing from 2-amino-5-hydroxybenzoic acid.3
0,34
7.3.6. 2-(Benzyloxy)-8,9-dihydro-6H-pyrido[2,1-b]quinazolin-
1
1(7H)-one (34)
7
3
.3.1. General synthetic procedure (I) for compounds 6, 7, 8, 33,
4 and 35
The respective lactame or benzolactame and 6-benzyloxyisatoic
Starting from piperidin-2-one (1.11 g, 11.17 mmol) and 6-ben-
zyloxyisatoic anhydride 5 (1.50 g, 5.58 mmol). The title compound
was obtained as a yellow oil (900 mg, 80% yield). R : 2.314 min,
+H) , m/z calcd: 307.1402; found:
307.1458. H NMR (300 MHz, CDCl ) d: 7.69 (d, J = 2.9 Hz, 1H),
t
+
anhydride 5 were placed in a 10 mL crimp-sealed thick-walled
glass tube equipped with a pressure sensor and a magnetic stirrer.
The sealed reaction tube was placed inside the cavity of a CEM Dis-
cover focused microwave synthesis system, operated at 200 °C
18 2 2
HRESIMS (C19H N O
1
3
7.52 (t, J = 6.8 Hz, 1H), 7.47–7.41 (m, 2H), 7.41–7.27 (m, 4H), 5.13
(s, 2H), 4.04 (t, J = 6.2 Hz, 2H), 2.94 (t, J = 6.6 Hz, 2H), 2.11–1.77
1
3
(
temperature monitored by a built-in infrared sensor), power
(m, 4H) ppm.
3
C NMR (75 MHz, CDCl ) d: 161.98, 156.93,
2
00 W, for 1 h. The residue was purified by column chromatogra-
152.64, 142.22, 136.42, 128.63, 128.14, 128.08, 127.67, 125.10,
121.04, 107.02, 70.43, 42.44, 31.75, 22.15, 19.39 ppm.
phy using dichloromethane/methanol (20:1) as eluent system to
afford the title compounds.
7
.3.7. 2-(Benzyloxy)-7,8,9,10-tetrahydroazepino[2,1-
7
.3.2. 8-(Benzyloxy)isoindolo[1,2-b]quinazolin-10(12H)-on (6)
b]quinazolin-12(6H)-one (35)
Starting from isoindolin-1-one (410 mg, 3.4 mmol) and 6-benzyl-
Starting from azepan-2-one (1.26 g, 11.17 mmol) and 6-benzyl-
oxyisatoic anhydride 5 (1.50 g, 5.58 mmol). The title compound
oxyisatoic anhydride 5 (915 mg, 3.4 mmol).The title compound was
obtained as a beige solid (740 mg, 64% yield). Mp = 161–163 °C. R
t
:
was obtained as a yellow solid (900 mg, 80% yield). Mp = 127–
+
+
2
3
7
2
1
1
1
.938 min, HRESIMS (C22
H
16
N
2
O
2
+H) , m/z calcd: 342. 1245; found:
42.1286. H NMR (300 MHz, CDCl ) d: 8.23 (d, J = 6.7 Hz, 1H),
.90–7.78 (m, 2H), 7.70–7.55 (m, 3H), 7.53–7.33 (m, 7H), 5.21 (s,
128 °C. R
t
:
2.683 min, HRESIMS (C20
20
H N
2
O
2
+H) , m/z calcd:
1
1
3
3
321.1603; found: 321.1602. H NMR (300 MHz, CDCl ) d: 7.69 (d,
J = 2.9 Hz, 1H), 7.52 (t, J = 6.9 Hz, 1H), 7.49–7.41 (m, 2H), 7.41–
7.27 (m, 4H), 5.13 (s, 2H), 4.36 (d, J = 5.5 Hz, 2H), 3.15–2.88 (m,
1
3
H), 5.18 (s, 2H) ppm.
3
C NMR (75 MHz, CDCl ) d: 158.94,
1
3
54.33, 151.29, 132.93, 132.91, 132.23, 132.00, 131.40, 130.59,
29.62, 129.02, 128.70, 128.25, 127.74, 126.25, 125.09, 123.52,
07.32, 70.58, 49.91 ppm.
3
2H), 1.82 (d, J = 4.1 Hz, 6H) ppm. C NMR (75 MHz, CDCl ) d:
161.74, 157.55, 157.19, 142.16, 136.41, 128.64, 128.43, 128.15,
127.66, 124.94, 120.90, 107.57, 70.44, 42.98, 37.52, 29.58, 28.14,
2
5.53 ppm.
7
.3.3. 10-(Benzyloxy)-5H-isoquinolino[1,2-b]quinazolin-8(6H)-
one (7)
Starting from 3,4-dihydroisoquinolin-1(2H)-one (0.5 g, 3.4 mmol)
7.3.8. General synthetic procedure (II) for compounds 9, 10, 11,
36–38
and 6-benzyloxyisatoic anhydride 5 (915 mg, 3.4 mmol). The title
To a mixture of the benzylated quinazolinones and 10% Pd-C
ethanol was added. The mixture was stirred at room temperature
and under an atmosphere of H for 4 h. The mixture then was fil-
2
tered over celit and then the solvent removed under reduced pres-
sure to afford the title phenolic compounds.
compound was obtained as a beige solid (870 mg, 72% yield).
+
Mp = 157–159 °C. R
calcd: 355.1402; found: 355.1441. H NMR (300 MHz, CDCl
(
t
: 3.393 min, HRESIMS (C23
H
18
N
2
O
2
+H) , m/z
1
3
) d: 8.45
d, J = 7.9 Hz, 1H), 7.78 (d, J = 2.9 Hz, 1H), 7.74 (d, J = 8.9 Hz, 1H),
7
.53–7.26 (m, 9H), 5.19 (s, 2H), 4.51–4.36 (m, 2H), 3.20–3.00 (m, 2H)
1
3
ppm. C NMR (75 MHz, CDCl
3
) d: 161.49, 157.45, 147.54, 136.74,
7.3.9. 8-Hydroxyisoindolo[1,2-b]quinazolin-10(12H)-one (9)
1
1
36.37, 131.42, 129.31, 128.68, 128.21, 127.75, 127.72, 127.65,
27.51, 125.18, 121.46, 107.40, 70.53, 39.78, 27.53 ppm.
Starting
from
8-(benzyloxy)isoindolo[1,2-b]quinazolin-
10(12H)-one 6 (700 mg, 2.06 mmol) and 10% Pd-C (80 mg). The
title compound was obtained as a yellow solid (450 mg, 87%).
+
7
.3.4. 11-(Benzyloxy)-6,7-dihydrobenzo[3,4]azepino[2,1-
Mp = 306–308 °C. R
t
: 1.72 min, HRESIMS (C15
H
10
N
2
O
2
+H) , m/z
1
b]quinazolin-9(5H)-one (8)
calcd: 251.0776; found: 251.0815. H NMR (300 MHz, DMSO) d:
10.12 (s, 1H), 8.03 (d, J = 7.5 Hz, 1H), 7.74 (t, J = 6.7 Hz, 1H), 7.72–
7.63 (m, 2H), 7.60 (t, J = 7.4 Hz, 1H), 7.52 (t, J = 5.2 Hz, 1H), 7.31
(dd, J = 8.8, 2.9 Hz, 1H), 5.14 (s, 2H) ppm. C NMR (75 MHz, DMSO)
d: 159.17, 155.85, 152.14, 142.10, 139.94, 132.28, 131.67, 128.68,
128.49, 123.98, 123.55, 122.23, 121.27, 108.78, 49.58 ppm.
Starting from 2,3,4,5-tetrahydro-1H-benzo[c]azepin-1-one
(
3
0.55 g, 3.4 mmol) and 6-benzyloxyisatoic anhydride 5 (915 mg,
.4 mmol). The title compound was obtained as a yellow thick oil
1
3
+
(
750 mg, 60% yield). R
m/z calcd: 369.1603; found: 369.1610. H NMR (300 MHz, CDCl
d: 7.90–7.82 (m, 1H), 7.80 (d, J = 2.9 Hz, 1H), 7.73 (d, J = 8.9 Hz,
t 20 2 2
: 3.298 min, HRESIMS (C24H N O +H) ,
1
3
)
1
2
H), 7.53–7.29 (m, 9H), 7.22 (dd, J = 12.2, 10.8 Hz, 1H), 5.20 (s,
H), 4.97–3.18 (m, 2H), 2.76 (t, J = 7.1 Hz, 2H), 2.48–2.06 (m, 2H)
7.3.10. 10-Hydroxy-5H-isoquinolino[1,2-b]quinazolin-8(6H)-
one (10)
1
3
ppm. C NMR (75 MHz, CDCl
3
) d: 161.46, 157.56, 154.39, 142.62,
Starting from 10-(benzyloxy)-5H-isoquinolino[1,2-b]quinazo-
lin-8(6H)-one 7 (800 mg, 2.26 mmol) and 10% Pd-C (80 mg). The
title compound was obtained as a yellow solid (530 mg, 89%).
1
1
2
37.92, 136.35, 134.96, 131.11, 129.34, 128.95, 128.69, 128.61,
28.22, 127.70, 127.48, 125.13, 121.20, 107.36, 70.53, 41.59,
9.78, 28.79 ppm.
+
1
Mp = 255–257 °C. ESI-MS: 264.7 m/z [M] . H NMR (400 MHz,

F. H. Darras et al. / Bioorg. Med. Chem. 22 (2014) 4867–4881
4875
DMSO) d: 10.03 (s, 1H), 8.31 (dd, J = 7.8, 1.2 Hz, 1H), 7.62 (d,
J = 8.8 Hz, 1H), 7.45 (m, 4H), 7.30 (dd, J = 8.8, 2.9 Hz, 1H), 4.36–
4
organic solvent was removed under reduced pressure. The resulting
crude product was purified by column chromatography using
dichloromethane/methanol (30:1) as eluent system to afford the
title compounds.
.18 (m, 2H), 3.09 (t, J = 6.4 Hz, 2H) ppm. 13C NMR (101 MHz,
DMSO) d: 160.18, 156.08, 146.27, 140.43, 137.10, 131.04, 129.23,
1
2
28.97, 127.63, 127.02, 126.82, 123.90, 121.31, 109.03, 39.15,
6.44 ppm.
7.3.16. 8-(3-Bromopropoxy)isoindolo[1,2-b]quinazolin-
1
0(12H)-one (12)
Starting from 1,3-dibromopropane (810 mg, 4.0 mmol), 8-
7
.3.11. 11-Hydroxy-6,7-dihydrobenzo[3,4]azepino[2,1-
b]quinazolin-9(5H)-one (11)
hydroxyisoindolo[1,2-b]quinazolin-10(12H)-one
9
(400 mg,
Starting from 11-(benzyloxy)-6,7-dihydrobenzo[3,4]azepi-
no[2,1-b]quinazolin-9(5H)-one 8 (700 mg, 1.9 mmol) and 10%
Pd-C (80 mg). The title compound was obtained as a yellow solid
1
.60 mmol), potassium carbonate (415 mg, 3.0 mmol), and cata-
lytic amount of potassium iodide. The title compound was
obtained as a yellow solid (440 mg, 74%). Mp = 133–134 °C. ESI-
MS: 372.8 m/z [M] . H NMR (300 MHz, CDCl
J = 10.4, 4.5 Hz, 1H), 7.80–7.70 (m, 2H), 7.67–7.51 (m, 3H), 7.40–
7
2
1
1
2
(
470 mg, 89%). Mp = 257–259 °C. R
t
: 2.015 min, HRESIMS (C17
+H) , m/z calcd: 279.1128; found: 279.1127.
H
14
+
1
3
) d: 8.12 (dd,
+
1
N
O
2 2
H NMR
(
300 MHz, DMSO) d: 10.16 (s, 1H), 7.75 (dd, J = 7.4, 1.2 Hz, 1H),
.32 (m, 1H), 5.13 (s, 2H), 4.31–4.07 (m, 2H), 3.76–3.27 (m, 2H),
.47–2.21 (m, 2H) ppm. C NMR (75 MHz, CDCl ) d: 158.15,
3
54.97, 150.91, 141.82, 137.00, 130.50, 129.74, 126.72, 126.63,
22.34, 121.23, 120.96, 119.08, 104.61, 63.64, 47.56, 29.97,
7.59 ppm.
7
.60 (d, J = 8.8 Hz, 1H), 7.55–7.38 (m, 3H), 7.37–7.25 (m, 2H),
.92 (s, 2H), 2.70 (t, J = 6.5 Hz, 2H), 2.18–2.07 (m, 2H) ppm.
C NMR (75 MHz, DMSO) d: 159.14, 155.16, 151.83, 139.56,
36.54, 133.67, 129.62, 127.97, 127.67, 127.40, 125.74, 122.76,
19.86, 107.90, 39.84, 27.81, 27.01 ppm.
13
3
1
3
1
1
7
.3.17. 10-(3-Bromopropoxy)-5H-isoquinolino[1,2-
7
.3.12. 7-Hydroxy-2,3-dihydropyrrolo[2,1-b]quinazolin-9(1H)-
b]quinazolin-8(6H)-one (13)
Starting from 1,3-dibromopropane (810 mg, 4.0 mmol), 10-
hydroxy-5H-isoquinolino[1,2-b]quinazolin-8(6H)-one 10 (500 mg,
one (36)
Starting from 7-(benzyloxy)-2,3-dihydropyrrolo[2,1-b]quinazo-
lin-9(1H)-one 33 (750 mg, 2.57 mmol) and 10% Pd-C (80 mg). The
title compound was obtained as a beige solid (450 mg, 87% yield).
Mp = 284–287 °C. H NMR (300 MHz, DMSO) d: 9.95 (s, 1H),
1
.89 mmol), potassium carbonate (415 mg, 3.0 mmol), and cata-
lytic amount of potassium iodide. The title compound was
obtained as a yellow solid (550 mg, 76%). Mp = 136–138 °C. R
1
t
:
7
.47 (d, J = 8.8 Hz, 1H), 7.39 (d, J = 2.8 Hz, 1H), 7.23 (dd, J = 8.8,
+
3
.278 min, HRESIMS (C19
H17BrN
2
O
2
+H) , m/z calcd: 385.0552;
) d: 8.54–8.39 (m, 1H),
.79–7.65 (m, 2H), 7.51–7.40 (m, 2H), 7.36 (dd, J = 8.9, 2.9 Hz,
2
.9 Hz, 1H,), 4.12–3.94 (m, 2H), 3.08–2.94 (m, 2H), 2.21–2.10
1
found: 385.0541. H NMR (300 MHz, CDCl
3
(
m, 2H) ppm.
7
1
2
H), 7.32–7.26 (m, 1H), 4.50–4.38 (m, 2H), 4.22 (m, 2H), 3.28 (m,
7
1
.3.13. 2-Hydroxy-8,9-dihydro-6H-pyrido[2,1-b]quinazolin-
1(7H)-one (37)
H), 3.11 (t, J = 6.4 Hz, 2H), 2.43–2.28 (m, 2H) ppm. 13C NMR
(
3
75 MHz, CDCl ) d: 150.96, 149.70, 147.64, 143.69, 140.25,
Starting
quinazolin-11(7H)-one 34 (750 mg, 2.45 mmol) and 10% Pd-C
80 mg). The title compound was obtained as a beige solid (430 mg,
from
2-(benzyloxy)-8,9-dihydro-6H-pyrido[2,1-b]
1
1
37.83, 136.78, 131.56, 129.16, 127.84, 127.70, 127.54, 124.91,
07.07, 67.50, 39.81, 32.84, 27.51, 6.36 ppm.
(
+
8
t 12 2 2
1%). Mp = 252–253 °C. R : 0.957 min, HRESIMS (C12H N O +H) ,
1
7.3.18. 11-(3-Bromopropoxy)-6,7-
m/z calcd: 217.0932; found: 217.0973. H NMR (300 MHz, DMSO)
d: 10.04 (s, 1H), 7.46 (d, J = 8.8 Hz, 1H), 7.39 (d, J = 2.8 Hz, 1H), 7.25
dihydrobenzo[3,4]azepino[2,1-b]quinazolin-9(5H)-one (14)
Starting from 1,3-dibromopropane (810 mg, 4.0 mmol), 11-
hydroxy-6,7-dihydrobenzo[3,4]azepino[2,1-b]quinazolin-9(5H)-
one 11 (555 mg, 2.0 mmol), potassium carbonate (415 mg,
(
dd, J = 8.8, 2.9 Hz, 1H), 3.92 (t, J = 6.1 Hz, 2H), 2.88 (t, J = 6.6 Hz,
H), 2.00–1.74 (m, 4H) ppm. 1 C NMR (75 MHz, DMSO) d: 160.52,
3
2
1
2
55.47, 152.63, 127.00, 123.89, 120.52, 108.63, 79.08, 41.71, 30.47,
1.28, 18.40 ppm.
3
.0 mmol), and catalytic amount of potassium iodide. The title
compound was obtained as beige solid (600 mg, 75%).
Mp = 138–140 °C. 3.184 min, HRESIMS (C20
m/z calcd: 399.0703; found: 399.0696. H NMR (300 MHz, CDCl
d: 7.91–7.81 (m, 1H), 7.76–7.65 (m, 2H), 7.52–7.32 (m, 3H), 7.24
a
+
t
R :
2 2
H19BrN O +H) ,
7
1
.3.14. 2-Hydroxy-7,8,9,10-tetrahydroazepino[2,1-b]quinazolin-
2(6H)-one (38)
1
3
)
Starting from 2-(benzyloxy)-7,8,9,10-tetrahydroazepino[2,1-b]-
(
(
d, J = 1.3 Hz, 1H), 4.27 (m, 2H), 3.53 (m, 2H), 3.36 (m, 2H), 2.76
t, J = 7.1 Hz, 2H), 2.26 (m, 4H) ppm. C NMR (75 MHz, CDCl
quinazolin-12(6H)-one 35 (700 mg, 3.04 mmol) and 10% Pd-C
13
3
) d:
(
(
80 mg). The title compound was obtained as a beige solid
14 2
430 mg, 81%). Mp = 249–251 °C. R : 1.268 min, HRESIMS (C13H N
1
1
3
61.33, 157.18, 152.63, 140.36, 138.44, 136.67, 129.73, 128.02,
27.30, 127.11, 126.75, 121.86, 119.06, 108.41, 41.60, 35.88,
5.38, 29.79, 28.78, 6.35 ppm.
t
+
1
O
2
+H) , m/z calcd: 231.1128; found: 231.1133. H NMR (400 MHz,
DMSO) d: 9.97 (s, 1H), 7.44 (d, J = 8.8 Hz, 1H), 7.38 (d, J = 2.8 Hz,
H), 7.22 (dd, J = 8.8, 2.9 Hz, 1H), 4.39–4.20 (m, 2H), 3.01 (dd,
1
7
.3.19. 7-(3-Bromopropoxy)-2,3-dihydropyrrolo[2,1-
13
J = 13.6, 5.9 Hz, 2H), 1.83–1.60 (m, 6H) ppm. C NMR (101 MHz,
DMSO) d: 160.62, 156.60, 155.68, 140.42, 128.16, 123.60, 120.51,
b]quinazolin-9(1H)-one (39)
Starting from 1,3-dibromopropane (810 mg, 4.0 mmol), 7-hydr-
oxy-2,3-dihydropyrrolo[2,1-b]quinazolin-9(1H)-one 36 (400 mg,
1
09.04, 41.83, 36.32, 28.69, 27.61, 25.01 ppm.
2
.0 mmol), potassium carbonate (415 mg, 3.0 mmol), and catalytic
7
1
.3.15. General synthetic procedure (III) for compounds 12, 13,
4, 39, 40 and 41
A mixture of 1,3-dibromopropane, free phenolic quinazolinone,
amount of potassium iodide. The title compound was obtained as a
yellow thick oil (430 mg, 67%). R : 2.135 min, HRESIMS (C14H15
BrN O +H) , m/z calcd: 323.0390; found: 323.0392. H NMR
2 2
t
+
1
potassium carbonate, and catalytic amount of potassium iodide
were refluxed in absolute acetonitrile for 2 h. After cooling the inor-
ganic salts were filtered off and the filtrate was concentrated under
reduced pressure. The remaining residue was taken up in brine, and
extracted with methylene chloride. The combined organic extracts
were washed with brine and dried over magnesium sulfate. The
(300 MHz, CDCl ) d: 7.55 (d, J = 2.5 Hz, 1H), 7.49 (dd, J = 8.9,
1.4 Hz, 1H), 7.29–7.16 (m, 1H), 4.23–4.02 (m, 4H), 3.66–3.47 (m,
3
2H), 3.08 (t, J = 7.9 Hz, 2H), 2.36–2.24 (m, 2H), 1.97 (m, 2H) ppm.
1
3
C NMR (75 MHz, CDCl ) d: 160.71, 157.41, 156.96, 143.67,
3
128.20, 124.38, 121.12, 106.71, 65.81, 46.50, 32.16, 32.14, 29.82,
19.59 ppm.

4
876
F. H. Darras et al. / Bioorg. Med. Chem. 22 (2014) 4867–4881
7
.3.20. 2-(3-Bromopropoxy)-8,9-dihydro-6H-pyrido[2,1-
t 27 3
(115 mg, 48%). Mp = 145–148 °C. R : 1.63 min, HRESIMS (C24H N
+
1
b]quinazolin-11(7H)-one (40)
O
2
+H) , m/z calcd: 390.2176; found: 390.2175. H NMR (300 MHz,
Starting from 1,3-dibromopropane (480 mg, 2.37 mmol), 2-
hydroxy-8,9-dihydro-6H-pyrido[2,1-b]quinazolin-11(7H)-one 37
CDCl ) d: 8.13 (dd, J = 10.4, 4.7 Hz, 1H), 7.74 (t, J = 6.0 Hz, 2H), 7.58
(m, 3H), 7.37 (dd, J = 8.9, 3.0 Hz, 1H), 5.15 (s, 2H), 4.16 (t, J = 6.2 Hz,
3
(
400 mg, 1.19 mmol), potassium carbonate (415 mg, 3.0 mmol),
2H), 2.93–2.70 (m, 6H), 2.21–2.03 (m, 2H), 1.83–1.54 (m, 8H) ppm.
C NMR (75 MHz, CDCl ) d: 160.50, 157.55, 153.08, 143.97,
3
1
3
and catalytic amount of potassium iodide. The title compound
was obtained as a beige solid (500 mg, 63%). Mp = 133–134 °C.
139.24, 132.85, 131.96, 128.94, 128.89, 124.61, 123.51, 123.20,
121.38, 106.93, 66.77, 55.40, 54.76, 49.80, 27.20, 27.01,
+
R
t
: 2.133 min, HRESIMS (C15
H
17BrN
2
O
2
+H) , m/z calcd: 337.0552;
1
found: 337.0548. H NMR (300 MHz, CDCl
3
) d: 7.63 (d, J = 2.9 Hz,
26.87 ppm. Elem. Anal. (C24
H, 6.53; Cl, 8.21; N, 9.73; O, 7.41 found: C, 68.10; H, 6.67; N, 9.65.
H
27
N
3
O
2
 0.5ÁCH
2 2
Cl ) calcd: C, 68.12;
1
4
2
H), 7.55 (d, J = 8.9 Hz, 1H), 7.35–7.27 (m, 1H), 4.18 (m, 2H),
.09 (t, J = 6.2 Hz, 2H), 3.69–3.33 (m, 2H), 2.99 (t, J = 6.6 Hz, 2H),
.48–2.24 (m, 2H), 2.10–1.87 (m, 4H) ppm. 13C NMR (75 MHz,
7.3.25. 8-(3-(Pyrrolidin-1-yl)propoxy)isoindolo[1,2-
b]quinazolin-10(12H)-one (17)
3
CDCl ) d: 156.93, 156.42, 155.87, 128.00, 125.35, 124.86, 124.31,
1
06.69, 65.85, 42.51, 32.23, 31.73, 29.82, 22.17, 19.38 ppm.
Starting from 8-(3-bromopropoxy)isoindolo[1,2-b]quinazolin-
1
0(12H)-one 12 (230 mg, 0.62 mmol) and pyrrolidine (66 mg,
0.93 mmol). The title compound was obtained as beige solid
23 3
(100 mg, 45%). Mp = 234–236 °C. R : 1.47 min, HRESIMS (C22H N
7
.3.21. 2-(3-Bromopropoxy)-7,8,9,10-tetrahydroazepino[2,1-
b]quinazolin-12(6H)-one (41)
t
+
1
Starting from 1,3-dibromopropane (480 mg, 2.37 mmol), 2-
hydroxy-7,8,9,10-tetrahydroazepino[2,1-b]quinazolin-12(6H)-one
O
2
+H) , m/z calcd: 362.1863; found: 362.1862. H NMR (300 MHz,
CDCl ) d: 8.13 (dd, J = 10.2, 4.6 Hz, 1H), 7.80–7.70 (m, 2H), 7.66–
3
3
8 (300 mg, 1.30 mmol), potassium carbonate (415 mg, 3.0 mmol),
and catalytic amount of potassium iodide. The title compound was
obtained as a beige solid (340 mg, 74%). Mp = 137–139 °C. R
7.52 (m, 3H), 7.38 (dd, J = 9.0, 2.9 Hz, 1H), 5.15 (s, 2H), 4.17 (t,
J = 6.4 Hz, 2H), 2.75–2.65 (m, 2H), 2.64–2.54 (m, 4H), 2.10 (m,
2H), 1.92–1.71 (m, 4H) ppm. C NMR (75 MHz, CDCl ) d: 160.50,
3
157.62, 153.04, 143.93, 139.24, 132.86, 131.93, 128.90, 128.87,
124.69, 123.49, 123.18, 121.37, 106.86, 66.96, 54.26, 53.11, 49.80,
1
3
t
:
+
2
.532 min, HRESIMS (C16
H
19BrN
2
O
2
+H) , m/z calcd: 351.0703;
) d: 7.61 (dd, J = 10.6,
.9 Hz, 2H), 7.36–7.28 (m, 1H), 4.41 (d, J = 5.5 Hz, 2H), 4.18 (m,
H), 3.71–3.55 (m, 2H), 3.08 (d, J = 6.4 Hz, 2H), 2.47–2.26 (m,
H), 1.97–1.75 (m, 6H) ppm. 13C NMR (75 MHz, CDCl
) d: 161.77,
1
found: 351.0706. H NMR (300 MHz, CDCl
3
5
2
2
1
4
28.62, 23.48 ppm. Elem. Anal. (C22
64.78; H, 5.84; Cl, 11.79; N, 9.98; O, 7.60 found: C, 64.81; H,
5.88; N, 10.15.
H
23
N
3
O
2
 0.7ÁCH
2 2
Cl ) calcd: C,
3
57.61, 157.16, 128.40, 124.67, 122.75, 120.22, 107.22, 65.88,
3.11, 32.77, 32.19, 29.79, 29.57, 28.09, 25.48 ppm.
7.3.26. 8-(3-(Diethylamino)propoxy)isoindolo[1,2-b]quinazolin-
1
0(12H)-one (18)
7
.3.22. General synthetic procedure (IV) of 3-substituted
Starting from 8-(3-bromopropoxy)isoindolo[1,2-b]quinazolin-
quinazolinones
10(12H)-one 12 (230 mg, 0.62 mmol) and diethylamine (68 mg,
The corresponding amine, 3-bromopropoxy quinazolinone,
potassium carbonate, and catalytic amount of potassium iodide
were refluxed in absolute acetonitrile for 3 h. After cooling the
inorganic salts were filtered off and the filtrate was concentrated
under reduced pressure. The remaining residue was taken up in
aq solution of NaCl, and extracted with methylene chloride. The
combined organic extracts were washed with brine and dried over
magnesium sulfate. The organic solvent was removed under
reduced pressure. The resulting crude product was purified by col-
umn chromatography using dichloromethane/methanol (15:1) as
eluent system to afford the corresponding target compounds:
0.93 mmol). The title compound was obtained as a beige solid
t 25
(100 mg, 44%). Mp = 120–123 °C. R : 1.508 min, HRESIMS (C22H
+
1
N
3
O
2
+H) , m/z calcd: 364.202; found: 364.2021.
H NMR
(300 MHz, CDCl ) d: 8.13 (dd, J = 10.2, 4.6 Hz, 1H), 7.79–7.70 (m,
3
2H), 7.65–7.50 (m, 3H), 7.43–7.32 (m, 1H), 5.15 (s, 2H), 4.15 (t,
J = 6.2 Hz, 2H), 2.72–2.64 (m, 2H), 2.59 (q, J = 7.2 Hz, 4H), 2.11–
1
3
3
1.89 (m, 2H), 1.06 (t, J = 7.2 Hz, 6H) ppm. C NMR (75 MHz, CDCl )
d: 160.51, 157.66, 153.02, 143.91, 139.24, 132.87, 131.93, 128.89,
128.87, 124.66, 123.49, 123.18, 121.38, 106.88, 66.91, 49.79,
49.34, 46.98, 26.75, 11.62 ppm. Elem. Anal. (C22
Cl ) calcd: C, 70.09; H, 6.73; Cl, 3.73; N, 11.05; O, 8.41 found: C,
0.02; H, 6.73; N, 10.82.
H
25
N
3
O
2
 0.2ÁCH
2
2
7
7
.3.23. 8-(3-(Piperidin-1-yl)propoxy)isoindolo[1,2-
b]quinazolin-10(12H)-one (15)
7.3.27. 8-(3-(3,4-Dihydroisoquinolin-2(1H)-
yl)propoxy)isoindolo[1,2-b]quinazolin-10(12H)-one (19)
Starting from 8-(3-bromopropoxy)isoindolo[1,2-b]quinazolin-
1
0
0(12H)-one 12 (230 mg, 0.62 mmol) and piperidine (80 mg,
.93 mmol). The title compound was obtained as beige solid
Starting from 8-(3-bromopropoxy)isoindolo[1,2-b]quinazolin-
10(12H)-one 12 (230 mg, 0.62 mmol) and 1,2,3,4-tetrahydroiso-
quinoline (124 mg, 0.93 mmol). The title compound was obtained
as beige solid (155 mg, 59%). Mp = 158–160 °C. ESI-MS: 424.0 m/z
(
t 25 3
105 mg, 45%). Mp = 193–196 °C. R : 1.555 min, HRESIMS (C23H N
+
1
O
CDCl
(
2
+H) , m/z calcd: 376.202; found: 376.2029. H NMR (300 MHz,
) d: 8.18–8.09 (m, 1H), 7.74 (dd, J = 5.9, 4.5 Hz, 2H), 7.66–7.52
m, 3H), 7.38 (dd, J = 8.9, 3.0 Hz, 1H), 5.15 (s, 2H), 4.13 (q, J = 6.7 Hz,
+
1
3
3
[MH] . H NMR (300 MHz, CDCl ) d: 8.15 (d, J = 6.9 Hz, 1H), 7.82–
7.70 (m, 2H), 7.67–7.51 (m, 3H), 7.40 (dd, J = 9.0, 2.9 Hz, 1H),
7.11 (d, J = 3.8 Hz, 5H), 5.14 (s, 2H), 4.19 (m, 2H), 3.65 (t,
J = 13.4 Hz, 4H), 2.94 (dd, J = 14.6, 10.6 Hz, 4H), 2.16 (m, 2H)
2
H), 2.50 (dd, J = 21.3, 13.5 Hz, 6H), 2.15–1.95 (m, 2H), 1.61
1
3
(
m, 4H), 1.53–1.37 (m, 2H) ppm. C NMR (75 MHz, CDCl
3
) d:
60.51, 157.67, 153.02, 143.91, 139.24, 132.87, 131.93, 128.89,
28.88, 124.74, 123.50, 123.18, 121.37, 106.80, 67.15, 55.96, 54.67,
9.80, 26.72, 25.92, 24.41 ppm. Elem. Anal. (C23
 0.06ÁCH
) calcd: C, 72.78; H, 6.65; Cl, 1.12; N, 11.04; O, 8.41 found: C,
3.02; H, 6.66; N, 10.85.
1
3
1
1
4
Cl
7
ppm.
3
C NMR (101 MHz, CDCl ) d: 160.47, 157.67, 153.04,
143.95, 139.27, 134.32, 134.24, 132.91, 131.92, 128.93, 128.86,
128.66, 126.63, 126.17, 125.64, 124.64, 123.49, 123.19, 121.42,
106.99, 66.93, 56.24, 54.92, 50.96, 49.79, 28.99, 27.06 ppm. Elem.
H N O
25 3 2
2
2
Anal. (C27
H
25
N
3
O
2
 0.1ÁCH
2 2
Cl ) calcd: C, 75.34; H, 5.88; Cl, 1.64;
N, 9.73; O, 7.41 found: C, 75.42; H, 5.89; N, 9.78.
7
1
.3.24. 8-(3-(Azepan-1-yl)propoxy)isoindolo[1,2-b]quinazolin-
0(12H)-one (16)
7.3.28. 8-(3-(1H-Imidazol-1-yl)propoxy)isoindolo[1,2-
b]quinazolin-10(12H)-one (20)
Starting from 8-(3-bromopropoxy)isoindolo[1,2-b]quinazolin-
10(12H)-one 12 (230 mg, 0.62 mmol) and imidazole (63 mg,
Starting from 8-(3-bromopropoxy)isoindolo[1,2-b]quinazolin-
0(12H)-one 12 (230 mg, 0.62 mmol) and azepane (90 mg,
.93 mmol). The title compound was obtained as a beige solid
1
0

F. H. Darras et al. / Bioorg. Med. Chem. 22 (2014) 4867–4881
4877
0
.93 mmol). The title compound was obtained as a beige solid
7.3.32. 10-(3-(Diethylamino)propoxy)-5H-isoquinolino[1,2-
b]quinazolin-8(6H)-one (24)
Starting from 10-(3-bromopropoxy)-5H-isoquinolino[1,2-b]
quinazolin-8(6H)-one 13 (240 mg, 0.62 mmol) and diethylamine
(68 mg, 0.93 mmol). The title compound was obtained as a beige
(
110 mg, 50%). Mp = 223–226 °C. R : 1.443 min, HRESIMS (C21
t
H
18
NMR
+
1
N
O
4 2
+H) , m/z calcd: 359.1503; found: 359.1504.
H
(
300 MHz, CDCl
3
) d: 8.20–8.12 (m, 1H), 7.78 (d, J = 9.0 Hz, 1H),
7
3
.71 (d, J = 2.9 Hz, 1H), 7.67–7.53 (m, 4H), 7.39 (dd, J = 9.0,
.0 Hz, 1H), 7.08 (s, 1H), 6.95 (s, 1H), 5.16 (s, 2H), 4.23 (t,
t
solid (130 mg, 56%). Mp = 153–155 °C. R : 1.771 min, HRESIMS
+
1
J = 6.9 Hz, 2H), 4.08 (t, J = 5.7 Hz, 2H), 2.40–2.22 (m, 2H) ppm.
(C23
27
H N
3
O
2
+H) , m/z calcd: 378.2176; found: 378.2172. H NMR
1
3
C NMR (75 MHz, CDCl
3
) d: 160.41, 156.94, 153.24, 144.33,
(300 MHz, CDCl ) d: 8.48–8.38 (m, 1H), 7.66 (dd, J = 8.6, 5.9 Hz,
3
1
1
4
39.27, 137.26, 132.78, 132.08, 130.34, 129.49, 129.18, 128.93,
24.39, 123.53, 123.26, 121.42, 119.01, 106.98, 64.47, 49.83,
2H), 7.47–7.37 (m, 2H), 7.33 (dt, J = 5.5, 2.8 Hz, 1H), 7.29–7.23
(m, 1H), 4.52–4.28 (m, 2H), 4.16 (m, 2H), 3.06 (m, 2H), 2.69 (dd,
J = 8.2, 6.7 Hz, 2H), 2.61 (q, J = 7.2 Hz, 4H), 2.11–1.93 (m, 2H),
3.62, 30.70 ppm. Elem. Anal. (C21
H
18
N
4
O
2
2 2
 0.4ÁCH Cl ) calcd:
1
3
C, 65.51; H, 4.83; Cl, 7.23; N, 14.28; O, 8.16 found: C, 65.42; H,
5
1.07 (t, J = 7.2 Hz, 6H) ppm. C NMR (75 MHz, CDCl
157.68, 147.32, 142.38, 136.69, 131.31, 129.72, 129.20, 127.65,
27.60, 127.48, 124.81, 121.47, 107.00, 66.80, 49.31, 46.96, 39.74,
2
27.52, 26.57, 11.48 ppm. Elem. Anal. (C23 Cl )
3
) d: 161.50,
.04; N, 14.40.
1
7
.3.29. 10-(3-(Piperidin-1-yl)propoxy)-5H-isoquinolino[1,2-
H
27
N
3
O
2
 0.25ÁCH
2
b]quinazolin-8(6H)-one (21)
Starting from 10-(3-bromopropoxy)-5H-isoquinolino[1,2-b]
quinazolin-8(6H)-one 13 (240 mg, 0.62 mmol) and piperidine
calcd: C, 70.04; H, 6.95; Cl, 4.45; N, 10.54; O, 8.03 found: C,
70.09; H, 6.90; N, 10.49.
(
80 mg, 0.93 mmol). The title compound was obtained as beige
7.3.33. 10-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)propoxy)-5H-
isoquinolino[1,2-b]quinazolin-8(6H)-one (25)
Starting from 10-(3-bromopropoxy)-5H-isoquinolino[1,2-b]
quinazolin-8(6H)-one 13 (240 mg, 0.62 mmol) and 1,2,3,4-tetrahy-
droisoquinoline (124 mg, 0.93 mmol). The title compound was
solid (145 mg, 60%). Mp = 127–129 °C. R : 1.814 min, HRESIMS
(
(
2
4
t
+
1
C
24
H
27
N
3
O
2
+H) , m/z calcd: 390.2176; found: 390.2188. H NMR
300 MHz, CDCl ) d: 8.43 (m, 1H), 7.67 (m, 2H), 7.49–7.37 (m,
H), 7.34 (d, J = 2.9 Hz, 1H), 7.27 (dd, J = 6.3, 2.2 Hz, 1H), 4.50–
.34 (m, 2H), 4.13 (m, 2H), 3.09 (m, 2H), 2.50 (m, 6H), 2.14–1.95
3
t
obtained as a beige solid (170 mg, 63%). Mp = 109–111 °C. R :
m, 2H), 1.61 (m, 4H), 1.53–1.32 (m, 2H) ppm. 13C NMR (75 MHz,
2.009 min, HRESIMS (C28
H
27
N
+H) , m/z calcd: 438.2176; found:
+
(
3
O
2
1
CDCl
3
) d: 161.52, 157.75, 147.31, 142.36, 136.70, 131.31, 129.74,
438.2172. H NMR (300 MHz, CDCl ) d: 8.48–8.39 (m, 1H), 7.75–
3
1
5
29.19, 127.66, 127.61, 127.49, 124.92, 121.46, 106.92, 67.12,
5.96, 54.65, 39.74, 27.53, 26.69, 25.90, 24.40 ppm. Elem. Anal.
7.64 (m, 2H), 7.48–7.40 (m, 2H), 7.40–7.31 (m, 2H), 7.31–7.24
(m, 2H), 7.11 (dd, J = 4.6, 3.3 Hz, 2H), 4.48–4.34 (m, 2H), 4.20 (t,
J = 6.3 Hz, 2H), 3.69 (s, 2H), 3.09 (t, J = 6.4 Hz, 2H), 2.93 (t,
J = 5.8 Hz, 2H), 2.79–2.70 (m, 4H), 2.22–2.09 (m, 2H) ppm.
(
C
24
H
27
N
3
O
2
2 2
 0.28ÁCH Cl ) calcd: 70.56; H, 6.72; Cl, 4.80; N,
1
0.17; O, 7.74 found: C, 70.75; H, 6.66; N, 9.97.
1
3
3
C NMR (75 MHz, CDCl ) d: 161.50, 160.50, 157.72, 147.34,
7
.3.30. 10-(3-(Azepan-1-yl)propoxy)-5H-isoquinolino[1,2-
142.40, 136.71, 134.25, 131.32, 129.22, 128.68, 127.67, 127.61,
127.50, 127.11, 126.62, 126.19, 125.65, 124.85, 121.49, 107.05,
66.87, 56.18, 54.91, 51.02, 39.75, 29.06, 27.53, 26.99 ppm. Elem.
b]quinazolin-8(6H)-one (22)
Starting from 10-(3-bromopropoxy)-5H-isoquinolino[1,2-b]
quinazolin-8(6H)-one 13 (240 mg, 0.62 mmol) and azepane
Anal. (C28
H
27
N
3
O
2
2 2
 0.15ÁCH Cl ) calcd: C, 75.09; H, 6.11; Cl,
(
92 mg, 0.93 mmol). The title compound was obtained as beige
2.36; N, 9.33; O, 7.11 found: C, 74.93; H, 6.11; N, 9.22.
solid (164 mg, 66%). Mp = 101–103 °C. R : 1.922 min, HRESIMS
t
+
1
(
(
2
(
C
25
H
29
N
3
O
2
+H) , m/z calcd: 404.2333; found: 404.2338. H NMR
300 MHz, CDCl ) d: 8.48–8.38 (m, 1H), 7.68 (dd, J = 9.0, 5.9 Hz,
H), 7.49–7.37 (m, 2H), 7.34 (dd, J = 8.9, 3.0 Hz, 1H), 7.30–7.25
m, 1H), 4.51–4.34 (m, 2H), 4.14 (t, J = 6.3 Hz, 2H), 3.09 (t,
J = 6.4 Hz, 2H), 2.80–2.63 (m, 6H), 2.14–1.94 (m, 2H), 1.76–1.52
7.3.34. 10-(3-(1H-Imidazol-1-yl)propoxy)-5H-isoquinolino[1,2-
b]quinazolin-8(6H)-one (26)
Starting from 10-(3-bromopropoxy)-5H-isoquinolino[1,2-b]
quinazolin-8(6H)-one 13 (240 mg, 0.62 mmol) and imidazole
(63 mg, 0.93 mmol). The title compound was obtained as beige solid
3
1
3
(
m, 8H) ppm.
3
C NMR (75 MHz, CDCl ) d: 161.53, 157.76,
t 20 4
(110 mg, 48%). Mp = 164–166 °C. R : 1.693 min, HRESIMS (C22H N
+
1
1
1
2
47.32, 142.37, 136.70, 131.32, 129.74, 129.21, 127.67, 127.62,
27.49, 124.89, 121.48, 106.97, 66.94, 55.50, 54.72, 39.75, 27.75,
7.54, 27.23, 27.02 ppm. Elem. Anal. (C25
O
2
+H) , m/z calcd: 373.1659; found: 373.1658. H NMR (300 MHz,
CDCl ) d: 8.50–8.37 (m, 1H), 7.76–7.69 (m, 1H), 7.62 (d, J = 2.9 Hz,
3
H
29
N
3
O
2
 0.15ÁCH
2
Cl
2
)
1H), 7.50 (s, 1H), 7.49–7.38 (m, 2H), 7.35 (dd, J = 8.9, 3.0 Hz, 1H),
7.31–7.24 (m, 1H), 7.06 (t, J = 0.9 Hz, 1H), 6.94 (t, J = 1.2 Hz, 1H),
4.45–4.36 (m, 2H), 4.21 (t, J = 6.9 Hz, 2H), 4.05 (t, J = 5.7 Hz, 2H),
calcd: C, 72.57; H, 7.09; Cl, 2.56; N, 10.09; O, 7.69 found: C,
7
2.62; H, 7.01; N, 9.87.
1
3
3
.09 (t, J = 6.4 Hz, 2H), 2.40–2.20 (m, 2H) ppm. C NMR (75 MHz,
7
.3.31. 10-(3-(Pyrrolidin-1-yl)propoxy)-5H-isoquinolino[1,2-
CDCl ) d: 161.44, 157.03, 147.65, 142.76, 137.27, 136.74, 135.12,
3
b]quinazolin-8(6H)-one (23)
Starting from 10-(3-bromopropoxy)-5H-isoquinolino[1,2-b]
quinazolin-8(6H)-one 13 (240 mg, 0.62 mmol) and pyrrolidine
131.46, 129.63, 129.47, 127.71, 127.65, 127.53, 124.58, 121.49,
119.00, 107.07, 64.44, 43.56, 39.79, 30.69, 27.49 ppm. Elem. Anal.
(C22
20
H N
4
O
2
3
 0.125ÁCHCl ) calcd: C, 68.61; H, 5.24; Cl, 3.43; N,
(
66 mg, 0.93 mmol). The title compound was obtained as a beige
14.46; O, 8.26 found: C, 68.24; H, 5.42; N, 14.85.
solid (150 mg, 64%). Mp = 124–125 °C. R : 1.730 min, HRESIMS
t
+
1
(
(
7
4
6
C
23
H
25
N
3
O
2
+H) , m/z calcd: 376.202; found: 376.2017. H NMR
300 MHz, CDCl ) d: 8.43 (m, 3H), 7.67 (dd, J = 9.3, 5.9 Hz, 6H),
.49–7.37 (m, 6H), 7.34 (dd, J = 8.9, 3.0 Hz, 3H), 7.30–7.25 (m,
7.3.35. 11-(3-(Piperidin-1-yl)propoxy)-6,7-
3
dihydrobenzo[3,4]azepino[2,1-b]quinazolin-9(5H)-one (27)
Starting from 11-(3-bromopropoxy)-6,7-dihydrobenzo[3,4]aze-
pino[2,1-b]quinazolin-9(5H)-one 14 (250 mg, 0.62 mmol) and piper-
H), 4.50–4.34 (m, 6H), 4.16 (t, J = 6.3 Hz, 6H), 3.12 (dt, J = 12.9,
.2 Hz, 8H), 2.85–2.72 (m, 8H), 2.66 (d, J = 5.5 Hz, 12H), 2.20–2.06
idine (80 mg, 0.93 mmol). The title compound was obtained as a
m, 6H), 1.92–1.79 (m, 12H) ppm. 13C NMR (75 MHz, CDCl
) d:
yellow oil (153 mg, 61%). R
m/z calcd: 404.2333; found: 404.2336. H NMR (300 MHz, CDCl
: 1.734 min, HRESIMS (C25
H
N
O
+H) ,
+
(
3
t
29
3
2
1
1
1
5
0
61.50, 157.60, 147.38, 142.44, 136.71, 131.34, 129.72, 129.24,
27.67, 127.62, 127.50, 124.82, 121.47, 107.03, 66.75, 54.22,
3
) d:
7.91–7.76 (m, 1H), 7.76–7.60 (m, 2H), 7.50–7.37 (m, 2H), 7.34 (dd,
J = 8.8, 3.0 Hz, 1H), 7.26–7.20 (m, 1H), 5.15–3.85 (m, 3H), 2.81–2.65
(m, 2H), 2.52–2.33 (m, 6H), 2.31–1.92 (m, 4H), 1.60 (m, 4H),
3.12, 39.75, 28.29, 27.53, 23.48 ppm. Elem. Anal. (C23
H
25
N
3
O
2
Â
.3ÁCH Cl ) calcd: C, 69.80; H, 6.44; Cl, 5.31; N, 10.48; O, 7.98
2
2
1
3
found: C, 69.72; H, 6.58; N, 10.56.
3
1.50–1.34 (m, 2H) ppm. C NMR (75 MHz, CDCl ) d: 161.49,

4
878
F. H. Darras et al. / Bioorg. Med. Chem. 22 (2014) 4867–4881
1
1
2
0
57.89, 154.14, 142.39, 137.89, 135.04, 131.03, 129.20, 128.90,
28.57, 127.44, 124.89, 121.18, 106.88, 67.14, 55.93, 54.66, 41.54,
9.78, 28.77, 26.70, 25.93, 24.41 ppm. Elem. Anal. (C25
.6ÁCH Cl ) calcd: C, 67.65; H, 6.70; Cl, 9.36; N, 9.25; O, 7.04 found:
2H), 7.16–7.07 (m, 4H), 4.91–3.90 (m, 4H), 3.68 (d, J = 13.5 Hz,
2H), 2.94 (t, J = 5.7 Hz, 2H), 2.74 (m, 6H), 2.15 (qd, J = 14.3, 6.2 Hz,
4H) ppm. C NMR (75 MHz, CDCl ) d: 161.47, 157.83, 154.20,
3
142.44, 137.91, 135.05, 134.41, 134.17, 131.04, 129.25, 128.92,
128.68, 128.59, 127.45, 126.62, 126.23, 125.69, 124.81, 121.21,
1
3
H N O
29 3 2
Â
2
2
C, 67.62; H, 6.73; N, 9.39.
1
07.01, 66.85, 56.10, 54.84, 50.98, 41.55, 29.78, 28.97, 28.77,
7
.3.36. 11-(3-(Azepan-1-yl)propoxy)-6,7-
26.92 ppm. Elem. Anal. (C29 Cl ) calcd: C, 71.72;
H, 6.12; Cl, 7.18; N, 8.51; O, 6.48 found: C, 71.46; H, 6.08; N, 8.47.
H
29
N
3
O
2
 0.5ÁCH
2
2
dihydrobenzo[3,4]azepino[2,1-b]quinazolin-9(5H)-one (28)
Starting from 11-(3-bromopropoxy)-6,7-dihydrobenzo[3,4]aze-
pino[2,1-b]quinazolin-9(5H)-one 14 (250 mg, 0.62 mmol) and aze-
7.3.40. 11-(3-(1H-Imidazol-1-yl)propoxy)-6,7-
pane (92 mg, 0.93 mmol). The title compound was obtained as a
dihydrobenzo[3,4]azepino[2,1-b]quinazolin-9(5H)-one (32)
Starting from 11-(3-bromopropoxy)-6,7-dihydrobenzo[3,4]aze-
pino[2,1-b]quinazolin-9(5H)-one 14 (250 mg, 0.62 mmol) and
imidazole (63 mg, 0.93 mmol). The title compound was obtained
+
yellow oil (170 mg, 66%). R
m/z calcd: 418.2489; found: 418.2495. H NMR (300 MHz, CDCl
d: 7.88–7.76 (m, 1H), 7.68 (t, J = 5.5 Hz, 2H), 7.51–7.38 (m, 2H),
t
: 1.833 min, HRESIMS (C26
H
31
N
3
O
2
+H) ,
1
3
)
7
.35 (dd, J = 8.9, 3.0 Hz, 1H), 7.24 (dd, J = 6.9, 1.7 Hz, 1H), 4.98–
.96 (m, 4H), 2.75 (dd, J = 6.7, 2.7 Hz, 7H), 2.22 (s, 2H), 2.14–2.01
t
as a yellow semi-solid. (110 mg, 48%). R : 1.636 min, HRESIMS (C23
+
1
3
22
H N
4
O
2
+H) , m/z calcd: 387.1816; found: 387.1812. H NMR
(
m, 2H), 1.79–1.54 (m, 10H) ppm. 13C NMR (75 MHz, CDCl
3
) d:
(300 MHz, CDCl ) d: 7.90–7.78 (m, 1H), 7.78–7.58 (m, 3H), 7.50–
3
1
1
5
61.49, 157.80, 154.20, 142.44, 137.90, 135.02, 131.05, 129.24,
28.90, 128.59, 127.45, 124.80, 121.19, 106.96, 66.82, 55.44,
5.21, 54.74, 41.55, 29.78, 28.77, 27.40, 27.00 ppm. Elem. Anal.
7.39 (m, 2H), 7.36 (dd, J = 8.9, 3.0 Hz, 1H), 7.11 (s, 1H), 6.95 (dd,
J = 5.9, 3.4 Hz, 1H), 5.40–4.48 (m, 2H), 4.27 (t, J = 6.8 Hz, 2H),
4.08 (t, J = 5.6 Hz, 2H), 2.75 (t, J = 6.9 Hz, 2H), 2.46–2.10 (m, 4H)
1
3
(
C
26
H
31
N
3
O
2
2
 0.6ÁCH Cl
2
) calcd: C, 68.19; H, 6.93; Cl, 9.08; N,
ppm. C NMR (75 MHz, CDCl
137.90, 137.07, 134.95, 131.14, 129.52, 128.92, 128.62, 128.50,
27.48, 124.51, 121.24, 119.19, 107.06, 64.42, 43.93, 41.61, 30.59,
29.77, 28.77 ppm. Elem. Anal. (C23 Cl ) calcd: C,
3
) d: 161.40, 157.10, 154.54, 142.81,
8
.97; O, 6.83 found: C, 67.93; H, 7.11; N, 9.17.
1
7
.3.37. 11-(3-(Pyrrolidin-1-yl)propoxy)-6,7-
H
22
N
4
O
2
 0.2ÁCH
2
2
dihydrobenzo[3,4]azepino[2,1-b]quinazolin-9(5H)-one (29)
Starting from 11-(3-bromopropoxy)-6,7-dihydrobenzo[3,4]aze-
pino[2,1-b]quinazolin-9(5H)-one 14 (250 mg, 0.62 mmol) and pyr-
69.07; H, 5.60; Cl, 3.52; N, 13.89; O, 7.93 found: C, 69.16; H,
5.68; N, 14.20.
rolidine (66 mg, 0.93 mmol). The title compound was obtained as a
7.3.41. 7-(3-(Piperidin-1-yl)propoxy)-2,3-dihydropyrrolo[2,1-
b]quinazolin-9(1H)-one (42)
Starting from 7-(3-bromopropoxy)-2,3-dihydropyrrolo[2,1-
b]quinazolin-9(1H)-one 39 (200 mg, 0.62 mmol) and piperidine
(80 mg, 0.93 mmol). The title compound was obtained as a white
+
brown oil (110 mg, 46%). R
m/z calcd: 390.2176; found: 390.2174. H NMR (300 MHz, CDCl
d: 7.89–7.80 (m, 1H), 7.69 (t, J = 6.0 Hz, 2H), 7.50–7.39 (m, 2H),
.35 (dd, J = 8.9, 3.0 Hz, 1H), 7.24 (dd, J = 3.9, 2.8 Hz, 1H), 4.18 (t,
J = 6.2 Hz, 4H), 2.96–2.65 (m, 8H), 2.21 (dd, J = 14.3, 6.6 Hz, 4H),
t
: 1.677 min, HRESIMS (C24
H
27
N
3
O
2
+H) ,
1
3
)
7
t
solid (120 mg, 59%). Mp = 123–125 °C. R : 1.1060 min, HRESIMS
1
3
+
1
1
1
1
2
.91 (s, 4H) ppm. C NMR (75 MHz, CDCl
3
) d: 161.47, 157.60,
(C19H
25
N
3
O
2
+H) , m/zm/z calcd: 328.202; found: 328.202. H NMR
54.63, 142.48, 137.90, 135.01, 131.07, 129.30, 128.91, 128.60,
27.46, 124.70, 121.18, 107.03, 66.54, 54.16, 53.12, 41.57, 29.78,
8.77, 27.89, 23.49 ppm. Elem. Anal. (C24
(300 MHz, CDCl ) d: 7.62 (d, J = 3 Hz, 1H), 7.55 (d, J = 9.0 Hz, 1H),
3
7.30 (m, 1H), 4.20 (t, J = 7.5 Hz, 2H), 4.11 (t, J = 6.3 Hz, 2H), 3.15
(t, J = 7.8 Hz, 2H), 2.64–2.48 (m, 6H) 2.22–2.33 (m, 2H), 2.14–2.05
(m, 2H), 1.61–1.73 (m, 4H), 1.42–1.52 (m, 2H) ppm. 13C NMR
H
27
N
3
O
2
2 2
 0.85ÁCH Cl )
calcd: C, 64.65; H, 6.27; Cl, 13.05; N, 9.10; O, 6.93 found: C,
6
4.61; H, 6.22; N, 9.00.
(75 MHz, CDCl
21.18, 107.10, 65.66, 55.35, 53.72, 46.53, 32.34, 24.39, 23.09,
22.42, 19.71 ppm. Elem. Anal. (C19 ) calcd: C,
50.22; H, 5.46; Cl, 29.09; N, 8.64; O, 6.58 found: C, 49.90; H, 5.82;
N, 8.93 ppm.
3
) d: 160.72, 157.48, 156.57, 144.03, 128.52, 124.02,
1
7
.3.38. 11-(3-(Diethylamino)propoxy)-6,7-
25
H N
3
O
2
 1.33ÁCHCl
3
dihydrobenzo[3,4]azepino[2,1-b]quinazolin-9(5H)-one (30)
Starting from 11-(3-bromopropoxy)-6,7-dihydrobenzo[3,4]aze-
pino[2,1-b]quinazolin-9(5H)-one 14 (250 mg, 0.62 mmol) and
diethylamine (68 mg, 0.93 mmol). The title compound was
7.3.42. 7-(3-(Azepan-1-yl)propoxy)-2,3-dihydropyrrolo[2,1-
b]quinazolin-9(1H)-one (43)
Starting from 7-(3-bromopropoxy)-2,3-dihydropyrrolo[2,1-b]
quinazolin-9(1H)-one 39 (200 mg, 0.62 mmol) and azepane
t
obtained as dark yellow oil (130 mg, 56%). R : 1.708 min, HRESIMS
+
1
(
(
C
24
H
29
N
3
O
2
+H) , m/z calcd: 392.2333; found: 392.2329. H NMR
300 MHz, CDCl ) d: 7.90–7.78 (m, 1H), 7.69 (t, J = 6.0 Hz, 2H),
.50–7.39 (m, 2H), 7.35 (dd, J = 8.9, 3.0 Hz, 1H), 7.24 (dd, J = 3.8,
.8 Hz, 1H), 4.78–3.77 (m, 4H), 2.91–2.59 (m, 8H), 2.38–1.98 (m,
H), 1.24–1.04 (m, 6H) ppm. 13C NMR (75 MHz, CDCl
) d: 161.48,
3
7
2
4
1
1
2
(92 mg, 0.93 mmol). The title compound was obtained as yellow
+
viscous oil (164 mg, 77%). R
m/z calcd: 342.2182; found: 342.2176. H NMR (300 MHz, CDCl
t
: 1.210 min, HRESIMS (C20
H
27
N
3
O
2
+H) ,
1
3
3
) d:
57.65, 154.28, 142.52, 137.90, 135.02, 131.07, 129.30, 128.91,
28.60, 127.46, 124.70, 121.22, 107.02, 66.54, 49.26, 46.94, 41.57,
9.78, 28.77, 26.08, 10.98 ppm. Elem. Anal. (C24
) calcd: C, 72.36; H, 7.36; Cl, 1.77; N, 10.51; O, 8.00 found: C,
2.18; H, 7.33; N, 10.18.
7.61 (d, J = 2.9 Hz, 1H), 7.54 (d, J = 8.9 Hz, 1H), 7.33–7.25 (m, 1H),
4.23–4.14 (m, 2H), 4.10 (t, J = 6.3 Hz, 2H), 3.21–3.05 (m, 2H),
2.76–2.61 (m, 6H), 2.36–2.19 (m, 2H), 2.07–1.92 (m, 2H), 1.61 (m,
29
H N
3
O
2
 0.1ÁCH
2
1
3
13
Cl
7
2
8H) ppm. C NMR (75 MHz, CDCl
160.85, 157.43, 157.11, 143.59, 128.20, 124.54, 121.18, 106.70,
6.91, 55.49, 54.68, 46.49, 36.49, 32.24, 31.42, 27.78, 27.24, 26.99,
19.68 ppm. Elem. Anal. (C20 ) calcd: C, 60.48;
3
)
3
C NMR (75 MHz, CDCl ) d:
6
7
.3.39. 11-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)propoxy)-6,7-
27
H N
3
O
2
 0.56ÁCHCl
3
dihydrobenzo[3,4]azepino[2,1-b]quinazolin-9(5H)-one (31)
H, 6.80; Cl, 14.59; N, 10.29; O, 7.84 found: C, 60.36; H, 6.95; N,
10.02.
Starting from 11-(3-bromopropoxy)-6,7-dihydrobenzo[3,4]aze-
pino[2,1-b]quinazolin-9(5H)-one 14 (250 mg, 0.62 mmol) and
1
,2,3,4-tetrahydroisoquinoline (124 mg, 0.93 mmol). The title com-
pound was obtained as a yellow oil (170 mg, 63%). R : 1.951 min,
+H) , m/z calcd: 452.2333; found: 452.2328.
H NMR (300 MHz, CDCl ) d: 7.84 (dd, J = 8.7, 4.0 Hz, 1H), 7.68
dd, J = 8.8, 5.0 Hz, 2H), 7.44 (pd, J = 7.5, 3.8 Hz, 2H), 7.30–7.21 (m,
7.3.43. 7-(3-(Pyrrolidin-1-yl)propoxy)-2,3-dihydropyrrolo[2,1-
b]quinazolin-9(1H)-one (44)
Starting from 7-(3-bromopropoxy)-2,3-dihydropyrrolo[2,1-b]
quinazolin-9(1H)-one 39 (200 mg, 0.62 mmol) and pyrrolidine
(66 mg, 0.93 mmol). The title compound was obtained as a yellow
t
+
HRESIMS (C29
H
29
N
3
O
2
1
3
(

F. H. Darras et al. / Bioorg. Med. Chem. 22 (2014) 4867–4881
4879
viscous oil (150 mg, 77%). ESI-MS. R
t
: 1.034 min, HRESIMS (C18
H
23-
(80 mg, 0.93 mmol). The title compound was obtained as beige
solid (135 mg, 64%). Mp = 120–122 °C. R : 1.098 min, HRESIMS
+
1
N
(
3
O
2
+H) , m/z calcd: 314.1863; found: 314.1871.
300 MHz, CDCl ) d: 7.58 (d, J = 2.9 Hz, 1H), 7.50 (t, J = 6.8 Hz,
H), 7.30–7.22 (m, 1H), 4.12 (m, 4H), 3.10 (dd, J = 10.4, 5.5 Hz,
H
NMR
t
+
1
3
(C20H
27
N
3
O
2
+H) , m/z calcd: 342.2176; found: 342.2178. H NMR
1
2
2
(300 MHz, CDCl ) d: 7.59 (t, J = 3.8 Hz, 1H), 7.50 (d, J = 8.9 Hz,
3
H), 2.76–2.67 (m, 2H), 2.67–2.58 (m, 4H), 2.34–2.17 (m, 2H),
1H), 7.33–7.23 (m, 1H), 4.08 (m, 4H), 2.96 (t, J = 6.6 Hz, 2H),
.17–2.01 (m, 2H), 1.86–1.76 (m, 4H) pm. 13C NMR (75 MHz,
2.55–2.33 (m, 6H), 2.07–1.87 (m, 6H), 1.59 (m, 4H), 1.50–1.35
1
3
CDCl
3
) d: 160.79 (NCN), 157.23, 157.19, 143.62, 128.20, 124.45,
21.14, 106.70, 66.69, 54.19, 53.07, 46.49, 32.22, 28.25, 23.44,
9.65 ppm. Elem. Anal. (C18 ) calcd: C, 62.94;
 0.3ÁCHCl
(m, 2H) ppm.
3
C NMR (75 MHz, CDCl ) d: 162.07, 157.28,
1
1
152.42, 141.98, 127.93, 124.93, 121.03, 106.55, 67.06, 55.95,
54.64, 42.44, 31.77, 26.69, 25.91, 24.40, 22.18, 19.42 ppm. Elem.
H
23
N
3
O
2
3
H, 6.73; Cl, 9.14; N, 12.03; O, 9.16 found: C, 62.69; H, 6.93; N, 12.15.
Anal. (C20
H
27
N
3
O
2
 0.25ÁCH
2 2
Cl ) calcd: C, 67.06; H, 7.64; Cl,
4
.89; N, 11.59; O, 8.82 found: C, 67.15; H, 7.65; N, 11.60.
7
.3.44. 7-(3-(Diethylamino)propoxy)-2,3-dihydropyrrolo[2,1-
7
.3.48. 2-(3-(Azepan-1-yl)propoxy)-8,9-dihydro-6H-pyrido[2,1-
b]quinazolin-9(1H)-one (45)
Starting from 7-(3-bromopropoxy)-2,3-dihydropyrrolo[2,1-
b]quinazolin-9(1H)-one 39 (200 mg, 0.62 mmol) and diethylamine
b]quinazolin-11(7H)-one (49)
Starting from 2-(3-bromopropoxy)-8,9-dihydro-6H-pyrido[2,1-
b]quinazolin-11(7H)-one 40 (210 mg, 0.62 mmol) and azepane
(
68 mg, 0.93 mmol). The title compound was obtained as a yellow
(
92 mg, 0.93 mmol). The title compound was obtained as beige
solid (150 mg, 62%). Mp = 78–80 °C. R 1.067 min, HRESIMS
t
:
+
1
solid (145 mg, 66%). Mp = 117–120 °C. R : 1.212 min, HRESIMS
t
(
C
18
H
25
N
3
O
2
+H) , m/z calcd: 316.202; found: 316.2023. H NMR
300 MHz, CDCl ) d: 7.62 (d, J = 2.9 Hz, 1H), 7.54 (d, J = 8.9 Hz,
H), 7.30 (dd, J = 8.9, 2.9 Hz, 1H), 4.22–4.14 (m, 2H), 4.10 (t,
J = 6.3 Hz, 2H), 3.13 (dd, J = 10.3, 5.5 Hz, 2H), 2.67–2.60 (m, 2H),
+
1
(
C
21
29
H N
3
O
2
+H) , m/z calcd: 356.2333; found: 356.2333. H NMR
300 MHz, CDCl ) d: 7.56 (d, J = 2.9 Hz, 1H), 7.47 (t, J = 5.8 Hz,
H), 7.27 (dd, J = 8.9, 2.9 Hz, 1H), 4.17–3.96 (m, 4H), 2.92 (q,
J = 6.5 Hz, 2H), 2.75–2.30 (m, 6H), 2.04–1.82 (m, 6H), 1.74–1.48
(
1
3
(
1
3
2
1
1
4
.55 (q, J = 7.1 Hz, 4H), 2.35–2.16 (m, 2H), 2.06–1.83 (m, 2H),
1
3
.02 (t, J = 7.1 Hz, 6H) ppm. 13C NMR (75 MHz, CDCl
) d: 160.86,
(m, 8H) ppm.
3
C NMR (75 MHz, CDCl ) d: 162.03, 157.29,
3
1
5
52.40, 141.94, 127.90, 124.88, 121.02, 106.56, 66.89, 55.48,
4.66, 42.42, 31.74, 27.84, 27.28, 26.98, 22.17, 19.40 ppm. Elem.
57.44, 157.10, 143.58, 128.20, 124.54, 121.18, 106.70, 66.87,
9.31, 46.97, 46.49, 32.24, 26.78, 19.68, 11.67 ppm. Elem. Anal.
Anal. (C21
H
29
3
N O
2
2 2
 0.35ÁCH Cl ) calcd: C, 66.57; H, 7.77; Cl,
(
C
18
H
25
N
3
O
2
 0.46 ethylacetate) calcd: C, 66.95; H, 8.12; N,
6
.44; N, 10.91; O, 8.31 found: C, 66.55; H, 7.67; N, 10.97.
1
1.81; O, 13.13 found: C, 66.57; H, 7.86; N, 12.26.
7
.3.49. 2-(3-(Pyrrolidin-1-yl)propoxy)-8,9-dihydro-6H-
7
.3.45. 7-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)propoxy)-2,3-
pyrido[2,1-b]quinazolin-11(7H)-one (50)
Starting from 72-(3-bromopropoxy)-8,9-dihydro-6H-pyr-
ido[2,1-b]quinazolin-11(7H)-one 40 (210 mg, 0.62 mmol) and pyr-
rolidine (66 mg, 0.93 mmol). The title compound was obtained as
beige solid (140 mg, 69%). Mp = 121–123 °C. ESI-MS.
dihydropyrrolo[2,1-b]quinazolin-9(1H)-one (46)
Starting from 7-(3-bromopropoxy)-2,3-dihydropyrrolo[2,1-b]
quinazolin-9(1H)-one 39 (200 mg, 0.62 mmol) and 1,2,3,4-tetrahy-
droisoquinoline (124 mg, 0.93 mmol). The title compound was
t
R :
obtained as a pale yellow solid (165 mg, 71%). Mp = 140–142 °C. R
t
:
+
1
3
7
4
2
.021 min, HRESIMS (C19
28.202. H NMR (300 MHz, CDCl ) d: 7.58 (d, J = 2.9 Hz, 1H),
H
25
N
3
O
2
+H) , m/z calcd: 328.202; found:
+
1
.357 min, HRESIMS (C23
H
25
N
3
O
2
+H) , m/z calcd: 376.202; found:
1
1
3
3
76.2023. H NMR (300 MHz, CDCl ) d: 7.63 (d, J = 2.9 Hz, 1H), 7.56
3
.49 (d, J = 8.9 Hz, 1H), 7.33–7.25 (m, 1H), 4.11 (t, J = 6.4 Hz, 2H),
.05 (t, J = 6.2 Hz, 2H), 2.95 (t, J = 6.6 Hz, 2H), 2.69–2.59 (m, 2H),
.58–2.46 (m, 4H), 2.10–1.86 (m, 6H), 1.83–1.71 (m, 4H) ppm.
(
d, J = 8.9 Hz, 1H), 7.32 (dd, J = 8.9, 2.9 Hz, 1H), 7.17–6.94 (m, 5H),
4
2
.26–4.10 (m, 2H), 3.66 (d, J = 4.0 Hz, 2H), 3.13 (dd, J = 10.4, 5.5 Hz,
H), 2.98–2.85 (m, 2H), 2.73 (m 4H), 2.34–2.01 (m, 4H). C NMR
) d: 160.83, 157.41, 157.14, 143.63, 134.71, 134.30,
1
3
13
C NMR (75 MHz, CDCl
27.92, 124.90, 121.03, 106.58, 66.93, 54.25, 53.09, 42.42, 31.76,
8.70, 23.45, 22.18, 19.41 ppm. Elem. Anal. (C19
.15ÁCH Cl ) calcd: C, 67.62; H, 7.50; Cl, 3.13; N, 12.35; O, 9.41
3
) d: 162.04, 157.25, 152.41, 141.98,
(
75 MHz, CDCl
3
1
2
0
1
6
28.66, 128.23, 126.59, 126.13, 125.61, 124.53, 121.21, 106.76,
6.85, 56.23, 54.92, 51.04, 46.49, 32.25, 29.13, 27.02, 19.67 ppm.
H
25
N
3
O
2
Â
2
2
Elem. Anal. (C23
H N O
25 3 2
 0.56 ethylacetate) calcd: C, 71.36; H,
found: C, 67.62; H, 7.37; N, 12.34.
6
.99; N, 9.89; O, 11.75 found: C, 71.12; H, 6.60; N, 9.97.
7
.3.50. 2-(3-(Diethylamino)propoxy)-8,9-dihydro-6H-
7
.3.46. 7-(3-(1H-Imidazol-1-yl)propoxy)-2,3-
pyrido[2,1-b]quinazolin-11(7H)-one (51)
dihydropyrrolo[2,1-b]quinazolin-9(1H)-one (47)
Starting from 7-(3-bromopropoxy)-2,3-dihydropyrrolo[2,1-b]
quinazolin-9(1H)-one 39 (200 mg, 0.62 mmol) and imidazole
Starting from 2-(3-bromopropoxy)-8,9-dihydro-6H-pyrido[2,1-
b]quinazolin-11(7H)-one 40 (210 mg, 0.62 mmol) and diethyl-
amine (68 mg, 0.93 mmol). The title compound was obtained as
(
65 mg, 0.93 mmol). The title compound was obtained as beige
beige solid (133 mg, 65%). Mp = 113–115 °C. R
t
: 1.074 min, HRE-
+H) , m/z calcd: 330.2176; found: 330.2178.
H NMR (300 MHz, CDCl ) d: 7.58 (d, J = 2.9 Hz, 1H), 7.49 (d,
J = 8.9 Hz, 1H), 7.28 (dd, J = 8.9, 2.9 Hz, 1H), 4.07 (dt, J = 10.4,
+
solid (145 mg, 75%). Mp = 158–160 °C. R : 1.137 min, HRESIMS
t
27 3 2
SIMS (C19H N O
+
1
1
(
C
17
H
18
N
4
O
2
+H) , m/z calcd: 311.1503; found: 311.1508. H NMR
3
(
300 MHz, CDCl ) d: 7.63–7.54 (m, 2H), 7.50 (s, 1H), 7.31 (dd,
3
J = 8.9, 3.0 Hz, 1H), 7.06 (s, 1H), 6.92 (s, 1H), 4.20 (m, 4H), 4.10–
6
6
1
4
.3 Hz, 4H), 2.94 (t, J = 6.6 Hz, 2H), 2.56 (m, 6H), 2.05–1.83 (m,
H), 1.00 (t, J = 7.1 Hz, 6H). C NMR (75 MHz, CDCl ) d: 162.05,
3
57.32, 152.37, 141.95, 127.91, 124.87, 121.04, 106.58, 66.90,
9.33, 46.98, 42.41, 31.75, 26.91, 22.18, 19.42, 11.80. Elem. Anal.
1
3
3
2
1
1
.92 (m, 2H), 3.17 (dd, J = 18.3, 10.5 Hz, 2H), 2.28 (qd, J = 7.9,
.5 Hz, 4H) ppm. C NMR (75 MHz, CDCl ) d: 160.75, 157.49,
3
56.74, 144.01, 137.28, 129.65, 128.52, 124.25, 121.26, 118.93,
06.82, 64.41, 46.53, 43.53, 32.28, 30.69, 19.67 ppm. Elem. Anal.
1
3
(
C
19
27
H N
3
O
2
2 2
 0.15ÁCH Cl ) calcd: C, 67.22; H, 8.04; Cl, 3.11; N,
(
C
17
H
18
N
4
O
2
 0.4 ethylacetate) calcd: C, 64.64; H, 6.18; N,
1
2.28; O, 9.35 found: C, 67.23; H, 8.04; N, 12.26.
1
6.21; O, 12.96 found: C, 64.95; H, 5.96; N, 16.05.
7
.3.51. 2-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)propoxy)-8,9-
7
.3.47. 2-(3-(Piperidin-1-yl)propoxy)-8,9-dihydro-6H-
dihydro-6H-pyrido[2,1b] quinazolin-11(7H)-one (52)
pyrido[2,1-b]quinazolin-11(7H)-one (48)
Starting from 2-(3-bromopropoxy)-8,9-dihydro-6H-pyrido[2,1-
b]quinazolin-11(7H)-one 40 (210 mg, 0.62 mmol) and piperidine
Starting from 2-(3-bromopropoxy)-8,9-dihydro-6H-pyrido[2,1-
b]quinazolin-11(7H)-one 40 (210 mg, 0.62 mmol) and 1,2,3,4-tet-
rahydroisoquinoline (124 mg, 0.93 mmol). The title compound

4
880
F. H. Darras et al. / Bioorg. Med. Chem. 22 (2014) 4867–4881
was obtained as beige solid (154 mg, 64%). Mp = 167–169 °C. R
t
:
7.3.55. 2-(3-(Pyrrolidin-1-yl)propoxy)-7,8,9,10-
+
1
.367 min, HRESIMS (C24
found: 390.2177. 1H NMR (300 MHz, CDCl
J = 2.9 Hz, 1H), 7.52 (d, J = 8.9 Hz, 1H), 7.34–7.27 (m, 1H), 7.14–
H
27
N
3
O
2
+H) , m/z calcd: 390.2176;
tetrahydroazepino[2,1-b]quinazolin-12(6H)-one (56)
Starting from 2-(3-bromopropoxy)-7,8,9,10-tetrahydroazepi-
no[2,1-b]quinazolin-12(6H)-one 41 (220 mg, 0.62 mmol) and
pyrrolidine (66 mg, 0.93 mmol). The title compound was obtained
3
)
d: 7.60 (d,
7
2
2
.05 (m, 3H), 7.00 (dd, J = 6.9, 4.3 Hz, 1H), 4.16 (t, J = 6.3 Hz,
H), 4.06 (t, J = 6.2 Hz, 2H), 3.67 (s, 2H), 2.94 (m, 4H), 2.85–
t
as yellow viscous oil (150 mg, 71%). ESI-MS. R : 1.239 min, HRE-
1
3
+
1
.67 (m, 4H), 2.21–2.06 (m, 2H), 2.06–1.84 (m, 4H) ppm.
) d: 162.04, 157.24, 152.46, 142.02, 134.61,
34.25, 128.66, 127.96, 126.59, 126.15, 125.62, 124.85, 121.06,
06.67, 66.81, 56.17, 54.90, 51.01, 42.43, 31.77, 29.06, 26.98,
2.18, 19.42 ppm. Elem. Anal. (C24 Cl ) calcd:
 0.2ÁCH
C
SIMS (C20
H
27
N
3
O
2
+H) , m/z calcd: 342.2176; found: 342.2181.
H
NMR (75 MHz, CDCl
3
NMR (300 MHz, CDCl
3
) d: 7.59 (d, J = 2.9 Hz, 1H), 7.54–7.47 (m,
1
1
2
1H), 7.28 (dd, J = 8.9, 2.9 Hz, 1H), 4.37 (d, J = 5.5 Hz, 2H), 4.11 (t,
J = 6.4 Hz, 2H), 3.08–2.96 (m, 2H), 2.64 (dd, J = 9.1, 5.9 Hz, 2H),
2.59–2.48 (m, 4H), 2.12–1.98 (m, 2H), 1.90–1.73 (m, 10H) ppm.
H
27
N
3
O
2
2
2
1
3
C, 71.51; H, 6.79; Cl, 3.49; N, 10.34; O, 7.87 found: C, 71.28; H,
C NMR (75 MHz, CDCl
128.29, 124.73, 120.89, 107.14, 66.92, 54.24, 53.08, 42.97, 37.55,
9.59, 28.65, 28.17, 25.54, 23.46 ppm. Elem. Anal. (C20
Cl ) calcd: C, 67.69; H, 7.71; Cl, 3.96; N, 11.72; O, 8.93
3
) d: 161.81, 157.50, 157.35, 141.93,
6
.62; N, 10.30.
2
27 3 2
H N O -
7
.3.52. 2-(3-(1H-Imidazol-1-yl)propoxy)-8,9-dihydro-6H-
 0.2ÁCH
2
2
pyrido[2,1-b]quinazolin-11(7H)-one (53)
found: C, 67.52; H, 7.69; N, 11.85.
Starting from 2-(3-bromopropoxy)-8,9-dihydro-6H-pyrido[2,1-
b]quinazolin-11(7H)-one 40 (210 mg, 0.62 mmol) and imidazole
7.3.56. 2-(3-(Diethylamino)propoxy)-7,8,9,10-
(
65 mg, 0.93 mmol). The title compound was obtained as a beige
tetrahydroazepino[2,1-b]quinazolin-12(6H)-one (57)
solid (130 mg, 65%). Mp = 105–108 °C. R : 1.006 min, HRESIMS
t
Starting from 2-(3-bromopropoxy)-7,8,9,10-tetrahydroazepi-
no[2,1-b]quinazolin-12(6H)-one 41 (220 mg, 0.62 mmol) and
diethylamine (68 mg, 0.93 mmol). The title compound was
+
1
(
(
7
1
2
C
18
H
20
N
4
O
2
+H) , m/z calcd: 325.1659; found: 325.166. H NMR
300 MHz, CDCl ) d: 7.50–7.43 (m, 2H), 7.42 (d, J = 5.7 Hz, 1H),
.23 (dd, J = 8.9, 2.9 Hz, 1H), 7.00–6.92 (m, 1H), 6.85 (t, J = 1.2 Hz,
H), 4.12 (t, J = 6.9 Hz, 2H), 4.06–3.89 (m, 4H), 2.89 (t, J = 6.6 Hz,
3
obtained as yellow viscous oil (142 mg, 67%). R
t
: 1.286 min, HRE-
+H) , m/z calcd: 344.2333; found: 344.2341.
H NMR (300 MHz, CDCl ) d: 7.58 (d, J = 2.9 Hz, 1H), 7.50 (d,
+
29 3 2
SIMS (C20H N O
13
1
H), 2.28–2.10 (m, 2H), 2.00–1.78 (m, 4H) ppm.
d: 161.85, 156.52, 152.76, 142.27, 137.23,
29.58, 128.14, 124.47, 121.00, 118.90, 106.66, 64.35, 43.45,
2.41, 31.70, 30.62, 22.08, 19.31 ppm. Elem. Anal. (C18
Cl ) calcd: C, 62.71; H, 5.93; Cl, 6.27; N, 15.98; O, 9.12
found: C, 62.76; H, 6.02; N, 15.91.
C
NMR
3
(
75 MHz, CDCl
3
)
J = 8.9 Hz, 1H), 7.33–7.23 (m, 1H), 4.37 (d, J = 5.4 Hz, 2H), 4.09 (t,
J = 6.3 Hz, 2H), 3.11–2.94 (m, 2H), 2.72–2.42 (m, 6H), 1.93 (m, 2H),
1
4
1
3
H
20
4
N O
2
-
3
1.81 (s, 6H), 1.01 (t, J = 7.1 Hz, 6H) ppm. C NMR (75 MHz, CDCl )
 0.31ÁCH
2
2
d: 161.81, 157.55, 157.31, 141.89, 128.26, 124.70, 120.89, 107.12,
66.88, 49.31, 46.97, 42.96, 37.53, 29.58, 28.16, 26.85, 25.54,
1
1.75 ppm. Elem. Anal. (C20
H
29
N
3
O
2
 0.2ÁCH
2 2
Cl ) calcd: C, 67.31;
7
.3.53. 2-(3-(Piperidin-1-yl)propoxy)-7,8,9,10-
H, 8.22; Cl, 3.93; N, 11.66; O, 8.88 found: C, 67.45; H, 8.33; N, 11.65.
tetrahydroazepino[2,1-b]quinazolin-12(6H)-one (54)
Starting from 2-(3-bromopropoxy)-7,8,9,10-tetrahydroazepi-
no[2,1-b]quinazolin-12(6H)-one 41 (220 mg, 0.62 mmol) and
piperidine (80 mg, 0.93 mmol). The title compound was obtained
7.3.57. 2-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)propoxy)-
7,8,9,10-tetrahydroazepino[2,1-b]quinazolin-12(6H)-one (58)
Starting from 2-(3-bromopropoxy)-7,8,9,10-tetrahydroazepi-
no[2,1-b]quinazolin-12(6H)-one 41 (220 mg, 0.62 mmol) and
1,2,3,4-tetrahydroisoquinoline (124 mg, 0.93 mmol). The title com-
t
as pale yellow viscous oil (145 mg, 66%). R : 1.307 min, HRESIMS
+
1
(
(
1
C
21
H
29
N
3
O
2
+H) , m/z calcd: 356.2333; found: 356.2338. H NMR
300 MHz, CDCl ) d: 7.59 (d, J = 2.9 Hz, 1H), 7.51 (d, J = 8.9 Hz,
H), 7.29 (dd, J = 8.9, 2.9 Hz, 1H), 4.38 (d, J = 5.5 Hz, 2H), 4.09 (t,
J = 6.4 Hz, 2H), 3.07–2.93 (m, 2H), 2.47 (dd, J = 20.0, 12.2 Hz, 6H),
.10–1.95 (m, 2H), 1.83 (s, 6H), 1.59 (m, 4H), 1.51–1.36 (m, 2H)
3
t
pound was obtained as a yellow thick oil (170 mg, 68%). R :
+
1.573 min, HRESIMS (C25
H
29
N
3
O
2
+H) , m/z calcd: 404.2333; found:
1
404.2338. H NMR (300 MHz, CDCl ) d: 7.61 (t, J = 3.2 Hz, 1H),
3
2
7.53 (dd, J = 8.9, 3.2 Hz, 1H), 7.34–7.27 (m, 1H), 7.17–6.98 (m,
4H), 4.33 (dd, J = 25.4, 5.0 Hz, 2H), 4.16 (t, J = 6.3 Hz, 2H), 3.67 (s,
2H), 3.08–2.86 (m, 4H), 2.83–2.66 (m, 4H), 2.10 (m, 2H), 1.80
1
3
ppm. C NMR (75 MHz, CDCl
3
) d: 161.83, 157.54, 157.34, 141.93,
28.29, 124.77, 120.89, 107.11, 67.08, 55.94, 54.64, 42.98, 37.55,
9.60, 28.17, 26.69, 25.91, 25.55, 24.40 ppm. Elem. Anal. (C21
Cl ) calcd: C, 66.80; H, 7.80; Cl, 6.10; N, 10.96; O,
1
2
N
1
3
H
29
3
(dd, J = 7.6, 4.5 Hz, 6H) ppm. C NMR (75 MHz, CDCl ) d: 161.80,
3
O
2
 0.33ÁCH
2
2
157.50, 157.38, 141.96, 134.60, 134.25, 128.66, 128.32, 126.60,
126.16, 125.63, 124.69, 120.92, 107.22, 66.81, 56.17, 54.89, 51.00,
8
.34 found: C, 66.80; H, 7.81; N, 10.84.
4
(C25
2.97, 37.55, 29.59, 29.06, 28.17, 26.97, 25.55 ppm. Elem. Anal.
Cl ) calcd: C, 71.98; H, 7.05; Cl, 3.37; N,
7
.3.54. 2-(3-(Azepan-1-yl)propoxy)-7,8,9,10-
H
29
N
3
O
2
 0.2ÁCH
2
2
tetrahydroazepino[2,1-b]quinazolin-12(6H)-one (55)
9.99; O, 7.61 found: C, 71.79; H, 6.92; N, 9.97.
Starting from 2-(3-bromopropoxy)-7,8,9,10-tetrahydroazepi-
no[2,1-b]quinazolin-12(6H)-one 41 (220 mg, 0.62 mmol) and aze-
pane (92 mg, 0.93 mmol). The title compound was obtained as a
7.3.58. 2-(3-(1H-Imidazol-1-yl)propoxy)-7,8,9,10-
tetrahydroazepino[2,1-b]quinazolin-12(6H)-one (59)
Starting from 2-(3-bromopropoxy)-7,8,9,10-tetrahydroazepi-
no[2,1-b]quinazolin-12(6H)-one 41 (220 mg, 0.62 mmol) and
imidazole (65 mg, 0.93 mmol). The title compound was obtained
t 31 3
yellow thick oil (153 mg, 67%). R : 1.420 min, HRESIMS (C22H N
+
1
O
2
+H) , m/z calcd: 370.2489; found: 370.2493. H NMR (300 MHz,
CDCl ) d: 7.59 (d, J = 2.9 Hz, 1H), 7.51 (d, J = 8.9 Hz, 1H), 7.29 (dd,
J = 8.9, 2.9 Hz, 1H), 4.38 (d, J = 5.5 Hz, 2H), 4.10 (t, J = 6.4 Hz, 2H),
3
as beige solid (138 mg, 66%). Mp = 117–120 °C. R
t
: 1.223 min, HRE-
+H) , m/z calcd: 339.1816; found: 339.1820.
H NMR (300 MHz, CDCl ) d: 7.58–7.50 (m, 2H), 7.46 (s, 1H),
+
3
6
1
5
2
6
8
.09–2.96 (m, 2H), 2.78–2.61 (m, 6H), 2.08–1.93 (m, 2H), 1.83 (s,
22 4 2
SIMS (C19H N O
H), 1.71–1.51 (m, 8H) ppm. 13C NMR (75 MHz, CDCl
1
3
) d: 161.83,
3
57.56, 157.33, 141.92, 128.29, 124.73, 120.90, 107.15, 66.92,
5.50, 54.68, 42.97, 37.55, 29.60, 28.17, 27.85, 27.28, 27.00,
7.32–7.23 (m, 1H), 7.03 (s, 1H), 6.93–6.84 (m, 1H), 4.46–4.28 (m,
2H), 4.18 (t, J = 6.9 Hz, 2H), 4.00 (t, J = 5.7 Hz, 2H), 3.10–2.95 (m,
1
3
5.55 ppm. Elem. Anal. (C22
8.99; H, 8.19; Cl, 3.67; N, 10.87; O, 8.28 found: C, 68.89; H,
.13; N, 10.98.
H
31
N
3
O
2
 0.2ÁCH
2
Cl
2
)
calcd: C,
2H), 2.33–2.18 (m, 2H), 1.91–1.72 (m, 6H) ppm. C NMR (75 MHz,
CDCl ) d: 161.71, 157.71, 156.82, 142.30, 137.29, 129.67, 128.57,
124.39, 120.94, 118.93, 107.25, 64.37, 43.49, 43.01, 37.54, 30.66,
3

F. H. Darras et al. / Bioorg. Med. Chem. 22 (2014) 4867–4881
4881
2
9.56, 28.12, 25.51 ppm. Elem. Anal. (C19
H
22
N
4
O
2
 0.2ÁCH
2
Cl
2
)
8. Chen, X.; Zenger, K.; Lupp, A.; Kling, B.; Heilmann, J.; Fleck, C.; Kraus, B.; Decker,
M. J. Med. Chem. 2012, 55, 5231.
calcd: C, 64.89; H, 6.35; Cl, 3.99; N, 15.76; O, 9.00 found: C,
6
9.
Fang, L.; Kraus, B.; Lehmann, J.; Heilmann, J.; Zhang, Y.; Decker, M. Bioorg. Med.
Chem. Lett. 2008, 18, 2905.
4.66; H, 6.41; N, 15.77.
1
1
0. Tumiatti, V.; Minarini, A.; Bolognesi, M. L.; Milelli, A.; Rosini, M.; Melchiorre, C.
Curr. Med. Chem. 2010, 17, 1825.
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7
1
.3.59. 8-(3-Cyclohexylpropoxy)isoindolo[1,2-b]quinazolin-
0(12H)-one (61)
(
3-Chloropropyl)cyclohexane (100 mg, 0.62 mmol), 8-hydrox-
12. Singh, M.; Kaur, M.; Kukreja, H.; Chugh, R.; Silakari, O.; Singh, D. Eur. J. Med.
Chem. 2013, 70, 165.
yisoindolo[1,2-b]quinazolin-10(12H)-one 9 (100 mg, 0.40 mmol),
potassium carbonate (150 mg, 0.75 mmol) and catalytic amount
of potassium iodide were refluxed in absolute acetonitrile
13. Pinho, B. R.; Ferreres, F.; Valentão, P.; Andrade, P. B. J. Pharm. Pharmacol. 2013,
6
5, 1681.
14. Konrath, E. L.; dos Santos Passos, C.; Klein-Júnior, L. C.; Henriques, A. T. J. Pharm.
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(
50 mL) for 24 h. After cooling, the inorganic salts were filtered
1
5. Macabeo, A. P.; Vidar, W. S.; Chen, X.; Decker, M.; Heilmann, J.; Wan, B.;
Franzblau, S. G.; Galvez, E. V.; Aguinaldo, M. A.; Cordell, G. A. Eur. J. Med. Chem.
2011, 46, 3118.
off and the filtrate was concentrated under reduced pressure. The
remaining residue was taken up in aq solution of NaCl, and
extracted with methylene chloride. The combined organic extracts
were washed with brine and dried over magnesium sulfate. The
organic solvent was removed under reduced pressure. The result-
ing crude product was subjected to column chromatography to
afford the title compound as a white solid (115 mg, 77%).
Mp = 144 –146 °C. ESI-MS: 375.0 m/z [MH] . H NMR (300 MHz,
CDCl ) d: 8.14 (d, J = 7.0 Hz, 1H), 7.81–7.65 (m, 2H), 7.56 (m 3H),
.38 (dd, J = 8.9, 4.5 Hz, 1H), 5.14 (s, 2H), 4.06 (t, J = 6.6 Hz, 2H),
1
1
6. Connolly, D. J.; Cusack, D.; O’Sullivan, T. P.; Guiry, P. J. Tetrahedron 2005, 61,
10153.
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18, 4786.
1
2
2
9. Rajput, R.; Mishra, A. Int. J. Pharm. Pharm. Sci. 2012, 4, 66.
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+
1
3
2
2
2. Vanparia, S.; Patel, T.; Dixit, R.; Dixit, B. Med. Chem. Res. 2013, 22, 5184.
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7
1
.92–1.60 (m, 7H), 1.44–1.08 (m, 6H), 1.01–0.79 (m, 2H) ppm.
C NMR (75 MHz, CDCl
1
3
3
) d: 160.49, 157.80, 152.95, 139.23,
24. El-Azab, A.; Abdel-Hamide, S.; Sayed-Ahmed, M.; Hassan, G.; El-Hadiyah, T.; Al-
Shabanah, O.; Al-Deeb, O.; El-Subbagh, H. Med. Chem. Res. 2013, 22, 2815.
1
1
2
39.23, 132.82, 131.92, 128.87, 128.82, 124.80, 123.47, 123.21,
21.33, 106.67, 69.01, 49.80, 37.43, 33.66, 33.34, 26.68, 26.52,
6.38 ppm.
Information about crystallization of compound 26 (CCDC No.
83183) and its crystallographic data can be found in the Support-
25. Testard, A.; Picot, L.; Lozach, O.; Blairvacq, M.; Meijer, L.; Murillo, L.; Piot, J.-M.;
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ing information.
2
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0. Darras, F. H.; Pockes, S.; Huang, G.; Wehle, S.; Strasser, A.; Wittmann, H.-J.;
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Acknowledgments
3
We gratefully acknowledge the German Science Foundation
DFG) for financial support (DFG DE 1546/6-1), as well as the
32. Tozer, M.; Kalindjian, B. Expert Opin. Ther. Pat. 2000, 10, 1045.
33. AbdulHameed, M.; Liu, J.; Pan, Y.; Fang, L.; Silva-Rivera, C.; Zhan, C.-G. Theor.
Chem. Acc. 2011, 130, 69.
(
German Academic Exchange Service (DAAD) for awarding a Ph.D.
scholarship to F.H. Darras and the German Academic National
Foundation (Studienstiftung des deutschen Volkes) for awarding
a Ph.D. scholarship to S. Wehle. Appreciation is expressed to
Professor S. Elz of Regensburg University for providing facilities
and lab space to F.H. Darras.
3
4. Roos, G. H. P.; Drastlik, K. A. Heterocycles 2003, 60, 2023.
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Los Rios, C.; Iriepa, I.; Moraleda, I.; Gonzalez-Lafuente, L.; Mendes, E.; Perez, C.;
Rodriguez-Franco, M. I.; Marco-Contelles, J.; Carmo Carreiras, M. ACS Chem.
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Supplementary data
3
3
4
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the online version, at http://dx.doi.org/10.1016/j.bmc.2014.06.045.
4
4
4
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