Focused library development of 2-phenylacrylamides as broad spectrum cytotoxic agents

Article abstract of DOI:10.1016/j.bmc.2012.10.003

With our lead compound (E)-3-(4-chlorophenyl)-2-(1H-pyrrole-2-carbonyl) acrylonitrile (1) inducing 50% growth inhibition of 11 cancer cell lines at 27-61 μM, potency enhancements were rapidly established through the synthesis of a series of focused compou

Full text of DOI:10.1016/j.bmc.2012.10.003

Bioorganic & Medicinal Chemistry 21 (2013) 333–347
Contents lists available at SciVerse ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Focused library development of 2-phenylacrylamides as broad spectrum
cytotoxic agents
Mark Tarleton a, Lauren Dyson a, Jayne Gilbert b, Jennette A. Sakoff b, Adam McCluskey a,
⇑
a Centre for Chemical Biology, Chemistry, The University of Newcastle, University Drive, Callaghan, NSW 2308, Australia
b Medical Oncology, Calvary Mater Newcastle, Edith Street, Waratah, NSW 2298, Australia
a r t i c l e i n f o
a b s t r a c t
Article history:
With our lead compound (E)-3-(4-chlorophenyl)-2-(1H-pyrrole-2-carbonyl)acrylonitrile (1) inducing 50%
growth inhibition of 11 cancer cell lines at 27–61 M, potency enhancements were rapidly established
Received 27 June 2012
Revised 19 September 2012
Accepted 3 October 2012
Available online 23 October 2012
l
through the synthesis of a series of focused compound libraries. Six highly focused libraries (46 compounds
in total) were synthesised. Each library allowed the identification of a new lead compound, viz Library A
identified (E)-3-(pentafluorophenyl)-2-(1H-pyrrole-2-carbonyl)acrylonitrile (11) and (E)-3-(1H-indol-3-
yl)-2-(1H-pyrrole-2-carbonyl)acrylonitrile (13) as inhibitors with improved cytotoxicity. Synthesis of
discrete libraries of amidoacrylamide analogues (Ar–C@C(CN)–ArzAr–C@C(CN)–C(@O)NH)–Ar) resulted
in a series of analogues significantly more potent that the lead, 1. Three furan three analogues: (E)-3-(5-
chlorofuran-2-yl)-2-cyano-N-(4-methoxybenzyl)acrylamide (33), (E)-3-(5-bromofuran-2-yl)-2-cyano-N-
(4-methoxybenzyl)acrylamide (34) and (E)-2-cyano-3-(furan-3-yl)-N-(4-methoxybenzyl)acrylamide
Keywords:
Focused library
Acrylamides
Cytotoxicity
Synthesis
(37) returned broad spectrum growth inhibition (GI50 values of 5–16 lM). Replacement of the furan moiety
with simple aromatics gave an additional three analogues: (E)-2-cyano-N-(4-methoxybenzyl)-3-phenyl-
acrylamide (39), (E)-3-(4-chlorophenyl)-2-cyano-N-(4-methoxybenzyl)acrylamide (41) and (E)-2-cyano-
N-(4-methoxyphenyl)-3-(naphthalen-1-yl)acrylamide (45) with GI50 values of 7–24 lM. The final library
retained the aromatic substituents but introduced a 3,4-dichlorbenzylamine moiety to afford the 1-naph-
thyl substituted 52, which was the most potent broad spectrum cytotoxic analogue produced here in with
an average GI50 = 8.6
lM. This represents a fivefold potency enhancement relative to 1 and a new cytotoxic
scaffold suitable for further development.
Ó 2012 Published by Elsevier Ltd.
1. Introduction
program facilitated the development of a family of (Z)-2-pheny-
lacrylonitriles. In this latter report we observed modest levels of
cytotoxicity with the related (E)-2-(1H-pyrrole-2-carbonyl)-3-
(aromatic) acrylonitriles such as 1 (Fig. 1).13
Over the past decade there has been an increasing prevalence in
the number of biologically active compounds that contain an
acrylonitrile moiety similar to those reported herein.14–23 The
acrylonitrile pharmacophore is active across multiple cellular
pathways known to be involved in the production of cytotoxic ef-
fects in cell line models of cancer. These pathways include tubulin
polymerisation inhibition,14–16,18 cell death via apoptosis,20–22 and
Cancer is a debilitating, life-threatening disease that will affect
1 in 3 people during the course of their lives.1 Current treatment
strategies for cancer have limited efficacy, especially in the com-
mon malignancies such as breast, colon and lung cancer. Since
the 1960’s cancer drug discovery development has provided about
100 approved products for the treatment of malignancy. While ma-
jor advances have been made in the chemotherapeutic manage-
ment of patients, one-half of all cancer patients either, do not
respond to therapy or relapse following initial response, and ulti-
mately die from their metastatic disease. Better targeted therapies
are clearly needed in the fight against cancer.
We have multiple research programs targeting novel modes of
action for anti-cancer drugs including protein phosphatase inhibi-
tors that accelerate the cell cycle leading to cell death and dynamin
inhibitors that affect cell cycle progression at the abscission
stage.2–12 Most recently our anti-cancer drug development
Figure 1. (E)-3-(4-Chlorophenyl)-2-(1H-pyrrole-2-carbonyl)acrylonitrile (1) inhib-
its the growth of eleven human cancer cell lines displaying a 31–84% growth
⇑
Corresponding author. Tel.: +61 249 216486; fax: +61 249 215472.
E-mail address: Adam.McCluskey@newcastle.edu.au (A. McCluskey).
inhibition at 100
examined were.
lM drug concentration. See Table 1 for details of cell lines
0968-0896/$ - see front matter Ó 2012 Published by Elsevier Ltd.
http://dx.doi.org/10.1016/j.bmc.2012.10.003

334
M. Tarleton et al. / Bioorg. Med. Chem. 21 (2013) 333–347
tyrosine kinase inhibition.17 Indeed it could be argued that our
leads are structurally related to the monomeric tyrphostins, which
are known kinase inhibitors. There are also reports of selectivity
between cytotoxicity and antifungal activity.19
be important and imparted favourable growth inhibition. Of the
other Library A analogues only the indole substituted 13 showed
even a moderate level of potency at the initial 25 lM concentra-
tion, and most notably a ꢀ50% growth inhibition against the
SW480, MCF-7 and BE2-C cell lines (Supplementary data, Table
S1). This level of growth inhibition while low was deemed encour-
aging enough to continue our studies through the development of
Library B.
Given the modest level potency, but apparent cell line specific-
ity, noted with (13) we chose to examine the effect of a simple pyr-
role to indole replacement through application of the same
chemistry (commencing with indole) and the same family of alde-
hydes as described in Scheme 1. This led to the synthesis of Library
B comprising the indolylcarbonylacrylamide analogues 14-23. The
growth inhibiton screening data is presented in Table 2.
Building on our medicinal chemistry paradigm of iterative fo-
cused library synthesis, biological screening, re-design and library
synthesis,2–13 we have identified (E)-3-(4-chlorophenyl)-2-(1H-
pyrrole-2-carbonyl)acrylonitrile (1) as a synthetically accessible
anti-cancer lead compound. Our lead acrylonitrile (1) was accessed
in two simple steps from pyrrole, cyanoacetic acid and p-chloro-
benzaldehyde. The commercial availability of a wide range of
aldehydes allowed for rapid exploration of structure activity rela-
tionships (SAR). Herein we report an exploration of the SAR associ-
ated with 1 via the synthesis and biological evaluation of a series of
focused compound libraries.
From Library B, three analogues, the 4-NO2 (18), pentafluoro
(21) and bis-indole (23) proceeded to GI50 determination. The
introduction of the indole nucleus saw a significant reduction in
cytotoxicity previously observed with the pyrroles (Table S2, Sup-
plementary data), cf. indoles 15 and 14 compared with pyrroles 1
and 5. As with the pyrrole analogue, the pentafluorophenyl
substituted 21 was the most active in this library, and we note that
the 4-NO2 substituted 18 was also active. As with Library A, we
note that electron withdrawing groups on the phenyl ring con-
ferred higher levels of cell growth inhibition compared with the
indolylacrylamide analogues of Library B.
We next turned our attention to the possible effect of introduc-
ing additional functionality to the linker chain with the synthesis
of Library C which introduced an amide moiety. In designing our
next library we combined the most favourable properties of the
pyrrole Library A with the 4-OCH3 and 3,4-dicholoro phenyl sub-
stituents that afforded high levels of activity in our original pheny-
lacrylonitrile compound series.13 Accordingly we set about
developing an expedient route to simple analogues that combined
theses characteristics with the additional requirement of introduc-
ing an amide moiety. Treatment of 4-methoxybenzylamine and
3,4-dichlorobenzylamine with methyl cyanoacetate afforded the
cyanoamides 25 and 26 in excellent yields. Subsequent treatment
of 25 and 26 with 1H-pyrrole-2-carbaldehyde and furfural afforded
Library C analogues 27–30 with the desired amide moiety in-
stalled.25–31
2. Results and discussion
Our focused compound library development commenced with
the synthesis of (E)-3-(4-chlorophenyl)-2-(1H-pyrrole-2-car-
bonyl)acrylonitrile (1). In a typical synthesis, cyanoacetic acid
was mixed with acetic anhydride followed by the addition of pyr-
role and the mixture heated at 75 °C for 35 min, extractive work up
and silica gel chromatography afforded 3-oxo-3-(1H-pyrrol-2-
yl)propanenitrile (4). Treatment of 4 in ethanol with p-chlorobenz-
aldehyde and a catalytic quantity of piperidine, after work up gave
1 in a 39% yield (two steps) (Scheme 1).24
In the development of our first focused library (Library A), 4 was
treated with ten aldehydes (see Table 1 for details),13,25–31 and the
resultant compounds (1, 5–13) were screened for growth inhibi-
tion against a panel of eleven cancer cell lines: HT29 and SW480
(colon carcinoma), MCF-7 (breast carcinoma), A2780 (ovarian car-
cinoma), H460 (lung carcinoma), A431 (skin carcinoma), DU145
(prostate carcinoma), BEC-2 (neuroblastoma), SJ-G2 (glioblas-
toma), MIA (pancreatic carcinoma) and SMA (spontaneous murine
astrocytoma).6 In vitro cytotoxicity assays have been used for dec-
ades as a tool to understand hypothesis driven questions regarding
drug action. Our efforts in this area do not include the use of nor-
mal cell lines as a single in vitro screening assay as normal cell
lines fail to provide a ‘go/no-go’ step in the drug development pro-
cess. Indeed, when used in a prospective manner, they have not
been highly predictive of in vivo toxicity.32 A single in vitro screen-
ing platform is unlikely to provide the data required to evaluate
risk and predict human toxicity. In this regard, it is widely accepted
that additional models using a tiered toxicity screening approach
are required in order to define and predict clinical toxicity. As part
of our drug development strategy small molecules that satisfy spe-
cific drugability characteristics as well as cytotoxicity will undergo
further biological evaluation in animal models.
Cytotoxicity screening of 27–30 (Library C, Table 3) gave rise to
a number of interesting outcomes. Firstly the 4-OCH3 furan ana-
logue 29 returned the highest level of activity noted thus far with
GI50 values of 16–33 lM. Secondly, analogues 27, 28 and 30
showed activity in specific cell lines rather than broad anti-cancer
activity across all cell lines. Analogue 27 returned GI50 values of 36
and 47
l
M for the MCF-7 and the BE2-C cell lines, respectively.
M). Pyrrole
Against all other cell lines 27 was inactive (GI50 >50
l
28 showed a similar cytotoxicity profile to 27, however, this
analogue displayed significant selectivity towards the MCF-7
breast cancer cell line; GI50 = 6 lM, six times more active than
Initial cytotoxicity efficacy was determined at 25 lM drug con-
centration. Two of the ten Library A analogues were sufficiently
active at (or near) our screening threshold (90% growth inhibition
the 4-OCH3 analogue, 27. This was the most potent analogue thus
far and represented a fivefold increase relative to the lead (1) and
15-fold MCF-7 selectivity. Of equal note was that the simple bio-
isosteric replacement of the pyrrole NH to the furan O atom gave
two analogues with good activity against the cell lines examined.
Furan 30 showed a similar selectivity profile to that observed with
at 25 lM drug concentration, see also Supplementary data Tables
S1–S6) to warrant the determination of GI50 values. These data
are presented in Table 1. The lead 4-Cl (1) and pentafluorophenyl
(11) returned average GI50 values of 40 and approximately
70
lM, respectively across the 11 cell lines examined. This sug-
gested that the presence of an electron withdrawing group may
Scheme 1. Reagents and conditions: (i) Ac2O, 85 °C, 5 min; (ii) RCHO (see table for details), piperidine (cat), EtOH reflux 2 h.

M. Tarleton et al. / Bioorg. Med. Chem. 21 (2013) 333–347
335
Table 1
Evaluation of the cytotoxicity (GI50
(l
M)) of Library A derivatives 1, and 5–13 against a panel of eleven cancer cell lines
Compound
HT29a
SW480a
MCF-7b
A2780c
H460d
A431e
DU145f
61 10
BE2-Cg
SJ-G2h
MIAi
SMAj
36
5
34
3
27
—
3
35
—
4
43
—
7
52 11
34
—
3
37
—
7
48 17
38
—
6
1
5
k
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
6
7
—
—
—
—
—
—
—
—
—
—
—
—
——
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
8
—
9
—
—
10
11
224
—
>100
—
23 14
64 24
31 10
64
—
9
>100
—
14
—
3
23 13
25
—
4
>100
—
—
—
—
—
12
—
43%l
56%l
—
—
—
—
49%l
—
—
—
13
a
b
c
d
e
f
HT29 and SW480 (colon carcinoma).
MCF-7 (breast carcinoma).
A2780 (ovarian carcinoma).
H460 (lung carcinoma).
A431 (skin carcinoma).
DU145 (prostate carcinoma).
BEC-2 (neuroblastoma).
g
h
i
SJ-G2 (glioblastoma).
MIA (pancreatic carcinoma).
SMA (spontaneous murine astrocytoma).
‘—’ = Insufficient growth inhibition at 25
j
k
lM drug concentrations to proceed to full dose response evaluation, percentage growth inhibition values are tabulated in
Table S1 (Supplementary data).
l
Percentage growth inhibition at 25
lM drug concentration.
28. It thus appeared that the 3,4-dicholorophenyl moiety played a
role in imparting cancer cell line specificity. Overall the furans 29
and 30 are 2- to 3-fold more potent than 1.
Given the potency we noted with the furans 29 and 30 we ex-
panded the Library C to developed a new focused library, Library
D, in which we retained the 4-OCH3Ph moiety of 29 as it had given

336
M. Tarleton et al. / Bioorg. Med. Chem. 21 (2013) 333–347
Table 2
Evaluation of the cytotoxicity (GI50
(l
M)) of Library B analogues 14–23 against a panel of eleven cancer cell lines
Compound
HT29a
SW480a
—
MCF-7b
—
A2780c
—
H460d
—
A431e
—
DU145f
—
BE2-Cg
—
SJ-G2h
—
MIAi
—
SMAj
—
k
—
14
15
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
16
17
—
—
—
—
—
—
—
—
—
—
46
—
—
41
—
1
42
33%
—
3
35
3
48
6
47
—
4
46
—
6
59
—
0
40
5
37
—
4
9
60
—
6
18
19
42%l
32%l
33%l
l
—
—
—
—
—
—
—
—
—
—
20
21
18
—
3
18
—
3
13
4
11
—
0
35
—
7
19
—
1
30
—
1
10
—
2
14
—
1
21
—
5
20
—
1
40%l
22
6.0 0.1
8.3 0.4
11
1
9.3 0.2
7.6 0.3
7.6 0.1
17
1
12
0
13
0
14
0
15
1
23
a
b
c
d
e
f
HT29 and SW480 (colon carcinoma).
MCF-7 (breast carcinoma).
A2780 (ovarian carcinoma).
H460 (lung carcinoma).
A431 (skin carcinoma).
DU145 (prostate carcinoma).
BEC-2 (neuroblastoma).
g
h
i
SJ-G2 (glioblastoma).
MIA (pancreatic carcinoma).
SMA (spontaneous murine astrocytoma).
‘—’ = Insufficient growth inhibition at 25
j
k
l
M drug concentrations to proceed to full dose response evaluation, percentage growth inhibition values are tabulated in
Table S2 (Supplementary data).
l
Percentage growth inhibition at 25
lM drug concentration.

M. Tarleton et al. / Bioorg. Med. Chem. 21 (2013) 333–347
337
Table 3
Evaluation of the cytotoxicity (GI50
(l
M)) of Library C analogues 27–30 against a panel of eleven cancer cell lines
Compound
HT29a
SW480a
MCF-7b
A2780c
H460d
A431e
DU145f
>50
BE2-Cg
SJ-G2h
>50
MIAi
>50
SMAj
>50
>50
>50
34
6
6
>50
>50
>50
47
2
4
4
27
28
29
85 12
>50
1
>50
>50
>50
>50
43
>50
>50
34
>50
22
20
5
6
31
2
27
20
6
5
24
4
32
1
16
1
29
24
1
7
24
31
1
3
33
38
2
7
>50
>50
>50
>50
>50
>50
>50
30
a
HT29 and SW480 (colon carcinoma).
MCF-7 (breast carcinoma).
A2780 (ovarian carcinoma).
H460 (lung carcinoma).
A431 (skin carcinoma).
DU145 (prostate carcinoma).
BEC-2 (neuroblastoma).
SJ-G2 (glioblastoma).
MIA (pancreatic carcinoma).
SMA (spontaneous murine astrocytoma).
b
c
d
e
f
g
h
i
j
Scheme 2. Reagents and conditions: (i) MeOH, MW, 200 W, 120 °C, 15 min; (ii) 1H-pyrrole-2-carbaldehyde or furfural, piperidine (cat), EtOH, MW, 200 W, 120 °C, 15 min.
rise to the broad spectrum cytotoxicity and modified the furan nu-
cleus. Library D was prepared as per Scheme 2, and screened for
cytotoxicity as before and these data are presented in Table 4.
On examination of the data presented in Table 4 it was immedi-
ately apparent that bulky or electron donating 5-substituted-
(furan-2yl)acrylamide derivatives (31, 32, and 35) lost the broad
spectrum activity of the furan lead, 29. Furan 31 was largely inac-
tive, but did show a toxicity profile similar to indole 13. Analogue
35 showed a 20-fold specificity towards the MCF-7 breast cancer
Having established that multiple variations in the furan moiety
were permitted we extended the scope of these studies and ex-
plored replacement of the furan ring with other simple aromatic
systems while retaining the 4-OCH3Ph moiety, using the same
chemistry as in Scheme 2 to afford Library E analogues 39-45,
which were subsequently screened against our panel of eleven
cancer cell lines. These data are presented in Table 5.
As can be seen from the data presented in Table 5, again the po-
sition and type of substituent on the newly introduced aromatic
ring had a pronounced effect on the observed cytotoxicity. Those
analogues containing a small electron withdrawing group or posi-
tioned the electron density at the C20/C30 region returned a signif-
icantly higher level of broad spectrum cytotoxicity than those
lacking such a substituent. This is particularly evident on compar-
ison of the C40-Cl (41) with the C40-OCH3 (43) with the former
cell line (GI50 = 3
1 lM). Fusing the phenyl ring in the form of
the benzofuran analogue 38 resulted in a loss of MCF-7 selectivity
with 38 inactive across all cell lines examined. Electron withdraw-
ing substituents were well tolerated with the 5-Cl (33) and 5-Br
(34) highly cytotoxic with average GI50 values of 12 and 13
respectively. Interestingly the 4-bromo (36) (cf. 5-bromo 34) saw
a marked reduction in potency from an average GI50 = 13 M to
P53 M, indicating a high level of substituent pattern dependence.
lM,
l
returning an average GI50 value of 11 lM, while the latter was
l
essentially inactive. Also of note again, was the positional depen-
This positional dependence was also apparent with the furan-3-
dence of a fused aromatic system such as the C10- and C20-naphthyl
ylacrylamide analogue (37) which was the most active analogue
analogues 45 and 44 with average GI50 values of P51 and 15 lM,
generated thus far with an average GI50 = 7
tent than the equivalent furan-2-ylacrylamide (29, Table 3).
l
M, four times more po-
respectively. The C1’ analogue 45 is roughly threefold more potent
than the corresponding C20 44. These findings are in keeping with

338
M. Tarleton et al. / Bioorg. Med. Chem. 21 (2013) 333–347
Table 4
Evaluation of the cytotoxicity(GI50
(
lM)) of Library D analogues 31–38 against a panel of eleven cancer cell lines
O
R
N
H
CN
O
Compound
HT29a
SW480a
MCF-7b
A2780c
60 11
H460d
>100
A431e
DU145f
BE2-Cg
SJ-G2h
MIAi
51
SMAj
92
6
29
—
2
21
—
1
58 11
39
—
6
21
—
4
75 14
4
69 14
31
32
33
34
k
—
—
—
—
—
—
—
9
1
13
14
0
1
9
1
9
1
15
16
0
1
9
9
1
1
12
12
1
1
11
13
0
1
13
13
1
0
15
16
0
0
14
14
0
0
10
1
8
11
2
12
0
56
36
>100
>100
3
1
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
35
8
20
7
2
28
6
14
4
61 17
36 11
23 0.3
65
8
3
36
37
6
1
8
0
1
6
0
8
0
5
0
7
0
7
0
8
0
0
7
0
—
—
40%
—
—
—
—
—
—
34%
—
38
a
b
c
d
e
f
HT29 and SW480 (colon carcinoma).
MCF-7 (breast carcinoma).
A2780 (ovarian carcinoma).
H460 (lung carcinoma).
A431 (skin carcinoma).
DU145 (prostate carcinoma).
BEC-2 (neuroblastoma).
SJ-G2 (glioblastoma).
MIA (pancreatic carcinoma).
SMA (spontaneous murine astrocytoma).
‘—’ = insufficient growth inhibition at 25
g
h
i
j
k
lM drug concentrations to proceed to full dose response evaluation, percentage growth inhibition values are tabulated in
Table S4 (Supplementary data).
the general trends observed with the substituted furan analogues
shown in Table 5.
3. Conclusion
From the data obtained thus far a pharmacophore of broad
spectrum cytotoxicity for these acrylamides has emerged that
has identified the acrylamide moiety as essential, but permits
the introduction of an amide spacer, considerably extending the
scope for the development of these analogues. This pharmacophore
was applied in the design and development of our final focused
library, Library F, in which the 3,4-dichlorophenyl moiety was
re-introduced. Note that with the parent pyrrole series this 3,4-
dichlorophenyl moiety afforded high levels of MCF-7 specificity,
but with this extended pharmacophore we anticipated more broad
spectrum activity would be observed. As anticipated, the data in
Table 6 shows four analogues (46, 48, 51 and 52) with average
Replacement of the 4-ClPh moiety of the lead 1, typically re-
moved all vestiges of activity against the panel of eleven cell lines
examined herein. Only the pentafluorophenyl substituted 11 saw
a similarly modest level of potency, but not across all the cell
lines examined. Pentafluoro 11 was inactive against the SW480,
DU145 and the SMA cell lines. Indole 13 showed modest
specificity towards the SW480, MCF-7 and BE2-C cell lines.
Replacement of the pyrrole moiety with an indole gave three ana-
logues; 18, 21 and 23; with average GI50 values of 46, 19 and
11
lM, respectively.
The introduction of an amide moiety was explored with the
synthesis of Library C, and this produced an immediate increase
in potency with the furan analogue 29 displaying good broad spec-
GI50 values ranging from ca. 10 to 70
matched that predicted, viz the most active analogue was
C1’-naphthyl (52), with an average GI50 value of 8.6 M.
lM. The trends in activity
trum activity with an average GI50 value of 27
lM. Within Library C
l
the pyrrole analogues displayed preferential activity towards the

M. Tarleton et al. / Bioorg. Med. Chem. 21 (2013) 333–347
339
Table 5
Evaluation of the cytotoxicity (GI50
(l
M)) of Library E analogues 39–45, against a panel of eleven cancer cell lines
Compound
HT29a
SW480a
MCF-7b
A2780c
H460d
A431e
DU145f
BE2-Cg
SJ-G2h
MIAi
24
SMAj
9
1
14
34
0
3
18
36
2
2
9
1
18
41
1
4
9
1
13
34
2
3
15
35
0
3
17
39
1
6
3
6
16
42
1
5
39
40
36
3
33
2
26
5
38
16
7
0
10
—
0
11
—
1
7
0
14
—
1
6
0
7
1
12
—
0
11
—
0
1
17
—
0
41
42
k
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
43
44
54
9
1
41
1
1
33
3
0
40
2
0
>100
37
3
1
80 16
42
2
2
40
4
1
49
8
1
43
4
0
13
14
16
16
15
0
15
16
0
14
15
15
15
45
a
b
c
d
e
f
HT29 and SW480 (colon carcinoma).
MCF-7 (breast carcinoma).
A2780 (ovarian carcinoma).
H460 (lung carcinoma).
A431 (skin carcinoma).
DU145 (prostate carcinoma).
BEC-2 (neuroblastoma).
SJ-G2 (glioblastoma).
MIA (pancreatic carcinoma).
SMA (spontaneous murine astrocytoma).
‘—’ = insufficient growth inhibition at 25
g
h
i
j
k
lM drug concentrations to proceed to full dose response evaluation, percentage growth inhibition values are tabulated in
Table S5 (Supplementary data).
MCF-7 cell line with pyrrole 27 a 34
l
M and 28 a 6
l
M potent
specificity towards the MCF-7 cell line (3 lM). The development
inhibitor of cell growth.
Exploration of the furan moiety with Library D afforded three
highly potent analogues. Furans 33, 34 and 37 were 12, 13 and
7 lM potent across the cell lines examined. The 3-furanyl 37,
was the most active furan analogue in this series. The phenyl
of the analogues herein clearly demonstrated that the cytotoxicity
of these acrylamide analogues was not derived solely from the
electron withdrawing effect, with other characteristics such as ori-
entation of electron density and lone electron pairs having a pro-
nounced effect.
substituted was MCF-7 specific returning the best activity yet
noted with GI50MCF-7 = 3 lM. Replacement of the furan moiety with
Library E gave five analogues that proceeded to GI50 determination.
The most noteworthy analogues in this library were the phenyl 39,
4-chlorphenyl 41 and 1-naphthyl 45 with average GI50 values of
While some of our molecules show potent in vitro cytotoxicity
in a range of cancer cell lines additional biological models will need
to be utilised in order to define the in vivo efficacy and toxicity of
these small molecules. These findings will be reported in due
course.
15, 11 and 15
of the 3,4-dichlorobenzyl moiety produced a further increase in
potency for the 1-naphthyl substituted 52 to 8.6 M.
Herein from six focused libraries comprising a total of 46 ana-
logues we have identified eleven analogues with GI50 <15 M; an
average of threefold more active than the lead. The most potent
analogue were 28 (6 M), 37 (7 M) and 52 (8.6 M). Of the
remaining analogues, the furan 35 displayed the highest level of
lM, respectively. In the final library introduction
4. Experimental section
4.1. Materials
l
l
All starting materials were purchased from Aldrich Chemical Co.
and Lancaster Synthesis. Solvents were bulk, and distilled from
glass prior to use. Reaction progress was monitored by TLC, on alu-
l
l
l

340
M. Tarleton et al. / Bioorg. Med. Chem. 21 (2013) 333–347
Table 6
Evaluation of the cytotoxicity (GI50
(l
M)) of Library F analogues 46–52 against a panel of eleven cancer cell lines
Compound
HT29a
SW480a
MCF-7b
A2780c
H460d
A431e
DU145f
71 15
BE2-Cg
SJ-G2h
MIAi
47
SMAj
70 15
49
—
7
29
—
5
49
—
6
61
—
6
43
—
6
32
—
1
43
—
2
2
43
—
8
46
47
k
—
—
—
33
—
1
30
—
0
21
—
1
32
—
1
33
—
0
30
—
0
30
—
1
31
—
0
30
—
0
31
1
33
—
2
48
49
—
>100
95
50
0
4
45
29
7
1
86 11
>100
>100
90
50
8
6
70
77
6
7
93
40
2
0
>100
79 11
>100
50
51
44
7
2
44
7
4
49
13
8
44
8
1
64 20
0
11
0
8
1
0
7
0
8
0
12
1
7
0
1
0
7
0
52
a
b
c
d
e
f
HT29 and SW480 (colon carcinoma).
MCF-7 (breast carcinoma).
A2780 (ovarian carcinoma).
H460 (lung carcinoma).
A431 (skin carcinoma).
DU145 (prostate carcinoma).
BEC-2 (neuroblastoma).
SJ-G2 (glioblastoma).
MIA (pancreatic carcinoma).
SMA (spontaneous murine astrocytoma).
‘—’ = insufficient growth inhibition at 25
g
h
i
j
k
lM drug concentrations to proceed to full dose response evaluation, percentage growth inhibition values are tabulated in
Table S2 (Supplementary data).
minium plates coated with silica gel with fluorescent indicator
(Merck 60 F254) and flash chromatography was conducted utiliz-
ing SNAP Biotage KP-SIL columns. 1H and 13C spectra were re-
corded on a Bruker Advance AMX 300 MHz spectrometer at
300.13 and 75.48 MHz, respectively. Chemical shifts are relative
to TMS as internal standard. All compounds returned satisfactory
mass spectra were obtained using a micromass liquid chromatog-
raphy Z-path (LCZ) platform spectrometer. Mass to charge ratios
(m/z) are stated with their peak intensity as a percentage in paren-
theses. All mass spectra were obtained via the ES method thus
fragmentation patterns were not observed. The University of
Wollongong, Australia, Biomolecular Mass Spectrometry
Laboratory, analyzed samples for HRMS. The spectra were run on
a micromass QTof2 spectrometer using polyethylene glycol or
polypropylene glycol as the internal standard. Compound purity
was confirmed by a combination of LC–MS (HPLC), micro and/or
high resolution mass spectrometry and NMR analysis. All
analogues are P95% purity.
4.1.1. Cell culture and stock solutions22,23
Stock solutions were prepared as follows and stored at ꢁ20 °C:
drugs were prepared as 40 mM solutions in DMSO. All cell lines
were cultured at 37 °C, under 5% CO2 in air and were maintained
in Dulbecco’s modified Eagle’s medium (Trace Biosciences,
Australia) supplemented with 10% foetal bovine serum, 10 mM
sodium bicarbonate penicillin (100
l
g mLꢁ1), streptomycin
(100
l
g mLꢁ1), and glutamine (4 mM).
4.1.2. In vitro growth inhibition assays
Cells in logarithmic growth were transferred to 96-well plates.
Cytotoxicity was determined by plating cells in duplicate in med-
ium (100
after plating), when the cells were in logarithmic growth, medium
(100 L) with or without the test agent was added to each well.
lL) at a density of 2500–4000 cells/well. On day 0 (24 h
l
After 72 h of drug exposure, growth inhibitory effects were evalu-
ated using the MTT (3-[4,5-dimethyltiazol-2-yl]-2,5-diphenyl-
tetrazolium bromide) assay and their absorbance was read at

M. Tarleton et al. / Bioorg. Med. Chem. 21 (2013) 333–347
341
540 nm. Percentage growth inhibition was determined at a fixed
drug concentration of 25 M. A value of 100% is indicative of total
13C NMR (DMSO-d6) (75 MHz): d 174.8, 153.2, 143.5, 130.6
(2 ꢂ Ar), 129.7 (2 ꢂ Ar), 129.3, 129.0, 127.9, 119.3, 117.7, 110.8,
107.8, 21.2;
l
cell growth inhibition. Those analogues showing appreciable per-
centage growth inhibition underwent further dose response analy-
sis to allow the calculation of GI50 values. The GI50 value is defined
as the drug concentration at which cell growth is 50 % inhibited
based on the difference between the optical density values on
day 0 and those at the end of drug exposure.33
IR (KBr) cmꢁ1: 3291 (NH), 2210 (CN), 1622 (C@O);
LRMS: (APCI M+1) 237. HRMS: Calcd for C15H12N2O; Exact
mass: 236.0950, found (ACPI M+1) 237.1028.
4.2.4. (E)-3-(4-Hydroxyphenyl)-2-(1H-pyrrole-2-carbonyl)acry
lonitrile (6)
4.2. Chemistry
Synthesized using the general procedure as for (1), from 3-oxo-
3-(1H-pyrrol-2-yl)propanenitrile and 4-hydroxybenzaldehyde to
afford (6) as an orange solid; 43%; 240–243 °C.
1H NMR (DMSO-d6) (300 MHz): d = 12.07 (br, NH), 8.68 (br, OH),
8.18 (s, 1H, CH@C), 7.99 (d, J = 8.7 Hz, 2H, Ar H2 + Ar H6), 7.28–7.23
(m, 2H, Pyr H5 + Pyr H3), 6.94 (d, J = 8.7 Hz, 2H, Ar H3 + Ar H5),
6.30–6.27 (m, 1H, Pyr H4);
4.2.1. General methods
THF was freshly distilled from sodium–benzophenone. Flash
chromatography was carried out using silica gel 200–400 mesh
(60 Å). 1H and 13CNMR were recorded at 300 and 75 MHz, respec-
tively using a Bruker Avance 300 MHz spectrometer in CDCl3, ace-
tone-d6 and DMSO-d6. GCMS was performed using a Shimadzu
GCMS-QP2100. The instrument uses a quadrupole mass spectrom-
eter and detects samples via electron impact ionization (EI). The
Children’s Medical Research Institute, Cell Signalling Unit Mass
Spectrometry Laboratory analyzed samples for HRMS. The spectra
were run on the VG Autospec-oa-tof tandem high resolution mass
spectrometer using CI (chemical ionization), with methane as the
carrier gas and PFK (perfluorokerosene) as the reference. All sam-
ples returned satisfactory analyses.
13C NMR (DMSO-d6) (75 MHz): d 175.0, 162.3, 153.3, 133.6
(2 ꢂ Ar), 129.2, 127.3, 123.1, 118.6, 118.4, 116.2 (2 ꢂ Ar), 110.5,
104.1;
IR (KBr) cmꢁ1: 3419 (OH), 3290 (NH), 2218 (CN), 1617 (C@O),
1603 (Ar);
LRMS: (APCI M+1) 239. HRMS: Calcd for C14H10N2O2; Exact
mass: 239.0829; found (ACPI M+1) 239.0829.
4.2.5. (E)-3-(4-Methoxyphenyl)-2-(1H-pyrrole-2-carbonyl)acry
lonitrile (7)
4.2.2. (E)-3-(4-Chlorophenyl)-2-(1H-pyrrole-2-
carbonyl)acrylonitrile (1)
Synthesized using the general procedure as for (1), from 3-oxo-
3-(1H-pyrrol-2-yl)propanenitrile and 4-methoxybenzaldehyde to
afford (7) as a yellow solid; 83%; 166–168 °C.
Cyanoacetic acid (1.360 g, 16 mmol) was added to Ac2O (8 mL)
and the resultant suspension was stirred and heated to 50 °C upon
which the solid material dissolved. Pyrrole (1.073 g, 16 mmol) was
then added and the solution was heated at 75 °C for 35 min. The
solution was then diluted with EtOAc (20 mL) and washed with
0.1 M NaOH (3 ꢂ 10 mL). The organic layer was then collected
and dried using MgSO4. The solvent was then removed under
vacuum and the residue purified by flash silica chromatography
(1:10 EtOAc/Hexanes to 1:1 EtOAc/Hexanes) to afford 3-oxo-3-
(1H-pyrrol-2-yl)propanenitrile, 70%. Next, to an ethanolic solution
(10 mL) of 4-methylbenzaldehyde (1.57 mmol) was added an eth-
anolic solution (10 mL) of 3-oxo-3-(1H-pyrrol-2-yl)propanenitrile
(1.49 mmol). This mixture was heated to 70 °C at which time,
piperidine (2 drops) was added, and the solution was then heated
under reflux for an additional 2 h. After this time, the solution was
cooled and the solvent removed in vacuo to afford a brown oil
which was purified by flash chromatography (1:10 EtOAc/Hex-
anes) to afford (1), as a yellow solid; 39%; 192–194 °C.
1H NMR (Acetone-d6) (300 MHz): d 11.30 (br, NH), 8.28 (s, 1H,
CH@C), 8.13–8.11 (m, 2H, Ar H2 + Ar H6), 7.66–7.63 (m, 2H, Ar
H3 + Ar H5), 7.45–7.44 (m, 1H, Pyr H5), 7.35–7.34 (m, 1H, Pyr
H3), 6.38–6.36 (m, 1H, Pyr H4);
1H NMR (DMSO-d6) (300 MHz): d 12.14 (br, NH), 8.24 (s, 1H,
CH@C), 8.08 (d, J = 8.9 Hz, 2H, Ar H2 + Ar H6), 7.28–7.25 (m, 2H,
Pyr H5 + Pyr H3), 7.13 (d, J = 8.9 Hz, 2H, Ar H3 + Ar H5), 6.31 (s,
1H, Pyr H4), 3.85 (s, 3H, OCH3);
13C NMR (DMSO-d6) (75 MHz): d 174.9, 163.0, 153.0, 133.1
(2 ꢂ Ar), 129.1, 127.5, 124.6, 118.9, 118.2, 114.8 (2 ꢂ Ar), 110.6,
105.5, 55.6;
IR (KBr) cmꢁ1: 3306 (NH), 2209 (CN), 1617 (C@O), 1507 (Ar);
LRMS: (APCI M+1) 253. HRMS: Calcd for C15H12N2O2; Exact
mass: 252.0899, found (ACPI M+1) 253.0979.
4.2.6. (E)-3-(4-Nitrophenyl)-2-(1H-pyrrole-2-carbonyl)acry
lonitrile (8)
Synthesized using the general procedure as for (1), from 3-oxo-
3-(1H-pyrrol-2-yl)propanenitrile and 4-nitrobenzaldehyde to af-
ford (8) as a purple solid; 37%; 199–200 °C.
1H NMR (DMSO-d6) (300 MHz): d 12.32 (br, NH), 8.40–8.38 (m,
3H, Ar H3 + Ar H5 + Pyr H5), 8.24–8.21 (m, 2H, Ar H2 + Ar H6), 7.24
(m, 2H, Pyr H5 + Pyr H3), 6.34 (s, 1H, Pyr H4);
13C NMR (DMSO-d6) (75 MHz): d 174.2, 150.6, 148.8, 138.2,
131.3 (2 ꢂ Ar), 130.6, 128.7, 124.0 (2 ꢂ Ar), 120.3, 116.6, 113.1,
111.1;
13C NMR (Acetone-d6) (75 MHz): d 173.6, 151.1, 137.4, 131.7
(2 ꢂ Ar), 130.8, 128.8 (2 ꢂ Ar), 126.9, 119.7, 118.7, 116.6, 110.4,
109.5;
IR (KBr) cmꢁ1: 3308 (NH), 2228 (CN), 1633 (C@O), 1517 (NO)
1343 (NO);
LRMS: (APCI M+1) 268. HRMS: Calcd for C14H9N3O3; Exact
mass: 267.0644, found (ACPI M+1) 268.0717.
IR (KBr) cmꢁ1: 3284 (NH), 2211 (CN), 1627 (C@O);
LRMS: (APCI M+1) 257. HRMS: Calcd for C14H9ClN2O: 256.0403,
found (ACPI M+1) 257.0478.
4.2.7. (E)-3-(3,4-Dichlorophenyl)-2-(1H-pyrrole-2-
carbonyl)acrylonitrile (9)
4.2.3. (E)-2-(1H-Pyrrole-2-carbonyl)-3-p-tolylacrylonitrile (5)
Synthesized using the general procedure as for (1), from 3-oxo-
3-(1H-pyrrol-2-yl)propanenitrile and 4-chlorobenzaldehyde to
afford (5) as a yellow solid; 81%; 242–244 °C.
Synthesized using the general procedure as for (1), from 3-oxo-
3-(1H-pyrrol-2-yl)propanenitrile and 3,4-dichlorobenzaldehyde to
afford (9) as a yellow solid; 66%; 178–181 °C.
1H NMR (DMSO-d6) (300 MHz): d 12.22 (br, NH), 8.24 (s, 1H,
CH@C), 7.95 (d, J = 8.0 Hz, 2H, Ar H2 + Ar H6), 7.37 (d, J = 8.0 Hz,
2H, Ar H3 + Ar H5), 7.29–7.26 (m, 2H, Pyr H3 + Pyr H5),
6.33–6.30 (m, 1H, Pyr H4), 2.37 (s, 3H, CH3);
1H NMR (Acetone-d6) (300 MHz): d 11.31 (br, NH), 8.29–8.27
(m, 2H, Ar H5 + CH@C), 8.11–8.08 (m, 1H, Ar H6), 7.84–7.81 (m,
1H, Ar H2), 7.45–7.36 (m, 2H, Pyr H5 + Pyr H3), 6.39–6.37 (m,
1H, Pyr H4);

342
M. Tarleton et al. / Bioorg. Med. Chem. 21 (2013) 333–347
13C NMR (Acetone-d6) (75 MHz): d 173.4, 149.6, 135.0, 132.4,
IR (KBr) cmꢁ1: 3260 (NH), 2212 (CN), 1611 (C@O);
LRMS: (ESI Mꢁ1) 260. HRMS: Calcd for C16H11N3O; Exact mass:
261.0902, found (ESI MꢁH) 260.0807.
132.1, 131.7, 130.8, 129.3, 127.2, 127.0, 119.0, 116.3, 111.0, 110.5;
IR (KBr) cmꢁ1: 3310 (NH), 2222 (CN), 1632 (C@O);
LRMS: (APCI M+1) 290. HRMS: Calcd for C14H8Cl2N2O; Exact
mass: 290.0014, found (ACPI M+1) 291.0079.
4.2.12. (E)-3-(4-Chlorophenyl)-2-(1H-indole-3-carbonyl)acrylo
nitrile (14)
4.2.8. (E)-2-(1H-Pyrrole-2-carbonyl)-3-(4-(trifluoromethyl)
phenyl)acrylonitrile (10)
Cyanoacetic acid (0.363, 4.26 mmol) was added to Ac2O
(8 mL) and the resultant solution was stirred and heated to
50 °C upon which the solid material dissolved. Indole (0.5 g,
4.26 mmol) was then added and the solution was heated at
85 °C for 5 min. The solution was cooled to 0 °C and the solid
was collected under suction and washed with ice cold MeOH
(2 ꢂ 5 mL) to afford 3-(1H-indol-3-yl)-3-oxopropanenitrile in a
76% yield. Next, to an ethanolic solution (10 mL) of 4-chloro-
benzaldehyde (1.14 mmol) was added an ethanolic solution
Synthesized using the general procedure as for (1), from 3-oxo-
3-(1H-pyrrol-2-yl)propanenitrile and 4-trifluoromethylbenzalde-
hyde to afford (10) as a yellow solid; 41%; 160–162 °C.
1H NMR (DMSO-d6) (300 MHz): d 12.30 (br, 1H), 8.37 (s, 1H,
CH@C), 8.19 (d, J = 8.1 Hz, 2H, Ar H2 + Ar H6), 7.94 (d, J = 8.1 Hz,
2H, Ar H3 + Ar H6), 7.32 (s, 2H, Pyr H5 + Pyr H3), 6.34 (s, 1H, Pyr
H4);
13C NMR (DMSO-d6) (75 MHz): d 174.4, 151.4, 136.0, 131.2,
130.8 (2 ꢂ Ar), 128.8, 128.5 (2 ꢂ Ar), 125.9, 125.8, 120.1, 116.8,
112.1, 111.0;
(10 mL)
of
3-oxo-3-(1H-indol-3-yl)-3-oxopropanenitrile
(1.08 mmol). This mixture was heated to 70 °C at which time,
piperidine (2 drops) was added, and the solution was then
heated under reflux for an additional 2 h. After this time, the
solution was cooled and the solvent removed in vacuo to yield
a yellow solid. The crude solid was then recrystallised from EtOH
to afford (14) as a yellow solid; 80%; 230–232 °C.
IR (KBr) cmꢁ1: 3295 (NH), 2228 (CN), 1637 (C@O), 1561 (Ar),
1325 (C–F);
LRMS: (ESI Mꢁ1) 289. HRMS: Calcd for C15H9F3N2O; Exact
mass: 290.0667, found (ESI MꢁH) 289.0673.
1H NMR (Acetone-d6) (300 MHz): d 11.31 (br, NH), 8.54 (s, 1H,
Ind H2), 8.34–8.30 (m, 1H, Ind H4), 8.22 (s, 1H, CH@C), 8.11 (d,
J = 8.7 Hz, 2H, Ar H2 + Ar H6), 7.66–7.57 (m, 3H, Ar H3 + Ar
H5 + Ind H7), 7.32–7.27 (m, 2H, Ind H5 + Ind H6);
4.2.9. (E)-3-(Pentafluorophenyl)-2-(1H-pyrrole-2-carbonyl)acry
lonitrile (11)
Synthesized using the general procedure as for (1), from 3-oxo-
3-(1H-pyrrol-2-yl)propanenitrile and 2,3,4,5-pentafluorobenzalde-
hyde to afford (11) as a white solid; 22%; 178–180 °C.
1H NMR (Acetone-d6) (300 MHz): d 11.01 (br, NH), 7.35–7.32
(m, 1H, Pyr H5), 7.22–7.19 (m, 1H, Pyr H3), 6.41–6.38 (m, 1H, Pyr
H4), 5.67 (s, 1H, CH@C);
13C NMR (Acetone-d6) (75 MHz): d 180.0, 149.8, 137.0, 136.3,
134.3, 131.4 (2 ꢂ Ar), 131.0, 128.7 (2 ꢂ Ar), 126.1, 123.2, 122.0,
121.4, 116.9, 113.9, 111.9, 111.6;
IR (KBr) cmꢁ1: 3175 (NH), 2211 (CN), 1636 (C@C), 1600 (C@O),
822 (Ar–Cl);
13C NMR (Acetone-d6) (75 MHz): d 174.3, 154.3, 142.5, 135.0,
134.7, 124.6 (2 ꢂ Ar), 120.2, 118.4, 114.7 (2 ꢂ Ar), 110.5, 106.3,
106.1;
LRMS: (ESI Mꢁ1) 305. HRMS: Calcd for C18H11ClN2O; Exact
mass: 306.0560, found (ESI MꢁH) 305.0533.
IR (KBr) cmꢁ1: 3367 (NH), 2209 (CN), 1656 (C@C), 1608 (C@O);
LRMS: ESI (Mꢁ1) 311. HRMS: Calcd for C14H5F5N2O; Exact
mass: 312.0322, found (ESI MꢁH) 311.1706.
4.2.13. (E)-2-(1H-Indole-3-carbonyl)-3-p-tolylacrylonitrile (15)
Synthesized using the general procedure as for (14), from 3-
oxo-3-(1H-indol-3-yl)-3-oxopropanenitrile and 4-chlorobenzalde-
hyde to afford (15) as a yellow solid; 88% 216–219 °C.
4.2.10. (E)-3-(Naphthalen-2-yl)-2-(1H-pyrrole-2-carbonyl)acry
lonitrile (12)
Synthesized using the general procedure as for (1), from 3-oxo-
3-(1H-pyrrol-2-yl)propanenitrile and 2-naphthaldehyde to afford
(12) as a brown solid; 35%; 140–142 °C.
1H NMR (DMSO-d6) (300 MHz): d 12.26 (br, NH), 8.43 (s, 1H, Ind
H2), 8.18–8.16 (m, 2H, Ind H4 + CH@C), 7.94 (d, J = 8.1 Hz, 2H, Ar
H2 + Ar H6), 7.55–7.52 (m, 1H, Ind H7), 7.38 (d, J = 8.1 Hz, 2H, Ar
H3 + Ar H5), 7.30–7.21 (m, 2H, Ind H5 + Ind H6), 2.37 (s, 3H, OCH3);
13C NMR (DMSO-d6) (75 MHz): d 181.3, 152.1, 142.9, 136.6,
135.7, 130.3 (2 ꢂ Ar), 129.7 (2 ꢂ Ar), 126.0, 123.4, 122.4, 122.3,
121.3, 117.8, 113.5, 112.4, 110.1, 21.2;
1H NMR (DMSO-d6) (300 MHz): d 12.27 (br, NH), 8.57 (s, 1H, Ar
H1), 8.44 (s, 1H, CH@C), 8.23–8.21 (m, 1H, Ar H5), 8.11–7.99 (m,
3H, Ar H3 + Ar H4 + Ar H8), 7.70–7.60 (m, 2H, Ar H7 + Pyr H5),
7.35–7.31 (m, 2H, Ar H6 + Pyr H3), 6.35–6.34 (m, 1H, Pyr H4);
13C NMR (DMSO-d6) (75 MHz): d 174.8, 153.2, 134.4, 133.4,
132.4, 129.7, 129.0, 128.9, 128.8, 128.7, 128.0, 127.7, 127.2,
124.7, 119.5, 119.2, 110.8, 109.1;
IR (KBr) cmꢁ1: 3262 (NH), 2216 (CN), 1636 (C@C), 1623 (C@O);
LRMS: (ESI M+1) 287. HRMS: Calcd C19H14N2O; Exact mass:
286.1106, found (ESI M+H) 287.1210.
IR (KBr) cmꢁ1: 3291 (NH), 2215 (CN), 1617 (C@O);
LRMS: (ACPI M+1) 273. HRMS: Calcd for C18H12N2O; Exact
mass: 272.0950, found (ACPI M+1) 273.1028.
4.2.14. (E)-3-(4-Hydroxyphenyl)-2-(1H-indole-3-carbonyl)acry
lonitrile (16)
Synthesized using the general procedure as for (14), from 3-
(1H-indol-3-yl)-3-oxopropanenitrile and 4-hydroxybenzaldehyde
to afford (16) as a yellow solid; 90%; 252–253 °C.
4.2.11. (E)-3-(1H-Indol-3-yl)-2-(1H-pyrrole-2-carbonyl)acry
lonitrile (13)
1H NMR (DMSO-d6) (300 MHz): d 12.17 (br, NH), 8.40 (s, 1H, Ind
H2), 8.17–8.13 (m, 2H, Ind H4 + CH@C), 7.98 (d, J = 8.8 Hz, 2H, Ar
H2 + Ar H6), 7.53–7.51 (m, 1H, Ind H7), 7.27–7.23 (m, 2H, Ind
H5 + Ind H6), 6.94 (d, J = 8.8 Hz, 2H, Ar H3 + Ar H5);
13C NMR (DMSO-d6) (75 MHz): d 181.5, 162.0, 152.3, 136.5,
135.0, 133.2 (2 ꢂ Ar), 126.1, 123.3, 122.1, 121.7, 121.3, 118.6,
116.1 (2 ꢂ Ar), 113.7, 112.3, 106.4;
Synthesized using the general procedure as for (1), from 3-oxo-
3-(1H-pyrrol-2-yl)propanenitrile and 1H-indole-3-carbaldehyde to
afford (13) as an orange solid; 95%; 300–302 °C.
1H NMR (DMSO-d6) (300 MHz): d 12.49 (br, NH), 12.00 (br, NH),
8.67 (s, 1H, CH@C), 8.63 (s, 1H, Ind H2), 7.92–7.89 (m, 1H, Ind H7),
7.59–7.56 (m, 1H, Ind H4), 7.36–7.20 (m, 4H, Ind H5 + Ind H6 + Pyr
H3 + Pyr H5), 6.31–6.29 (m, 1H, Pyr H4);
IR (KBr) cmꢁ1: 3351 (OH), 3232 (NH), 1606 (C@O);
LRMS: (ESI M+1) 289. HRMS: Calcd for C18H12N2O2; Exact mass:
288.0899, found (ESI M+H) 289.0929
13C NMR (DMSO-d6) (75 MHz): d 174.4, 145.7, 136.2, 131.9,
129.7, 127.2, 126.4, 123.5, 121.9, 120.3, 118.4, 117.5, 112.8,
110.4, 110.3, 99.4;

M. Tarleton et al. / Bioorg. Med. Chem. 21 (2013) 333–347
343
4.2.15. (E)-2-(1H-Indole-3-carbonyl)-3-(4-methoxyphenyl)acry
lonitrile (17)
4.2.19. (E)-2-(1H-Indole-3-carbonyl)-3-(perfluorophenyl)acrylo
nitrile (21)
Synthesized using the general procedure as for (14), from 3-
(1H-indol-3-yl)-3-oxopropanenitrile and 4-methoxybenzaldehyde
to afford (17) as a yellow solid; 92%; 256–258 °C.
1H NMR (DMSO-d6) (300 MHz): d 12.21 (br, NH), 8.42 (s, 1H, Ind
H2), 8.18–8.16 (m, 2H, Ind H4 + CH@C), 8.07 (d, J = 8.9 Hz, 2H, Ar
H2 + Ar H6), 7.54–7.52 (m, 1H, Ind H7), 7.26–7.24 (m, 2H, Ind
H5 + Ind H6), 7.14 (d, J = 8.9 Hz, 2H, Ar H3 + Ar H6), 3.86 (s, 3H,
OCH3);
Synthesized using the general procedure as for (14), from 3-
(1H-indol-3-yl)-3-oxopropanenitrile and 2,3,4,5,6-pentafluoro-
benzaldehyde to afford (21) as a yellow solid; 71%; 240–242 °C.
1H NMR (DMSO-d6) (300 MHz): d 12.18 (br, NH), 8.39-8.38 (m,
1H, Ind H2), 7.97–7.94 (m, 1H, Ind H4), 7.56–7.52 (m, 1H, Ind H7),
7.31–7.17 (m, 3H, Ind H5 + Ind H6 + CH@C);
13C NMR (DMSO-d6) (75 MHz): d 159.9, 138.2, 136.8, 136.0,
134.9, 130.4, 124.5, 123.9, 122.9, 122.7 (2 ꢂ Ar), 121.5 (2 ꢂ Ar),
121.2, 120.6, 119.2, 112.6, 105.1;
13C NMR (DMSO-d6) (75 MHz): d 181.4, 162.6, 151.9, 136.5,
135.3, 132.7 (2 ꢂ Ar), 126.2, 124.8, 123.4, 122.2, 121.3, 118.3,
114.7 (2 ꢂ Ar), 113.6, 112.3, 107.9, 55.6;
IR (KBr) cmꢁ1: 3283 (NH), 2212 (CN), 1655 (C@C), 1596 (C@O),
1207 (C–F);
IR (KBr) cmꢁ1: 3220 (NH), 2221 (CN), 1591 (C@O), 1176 (C–O);
LRMS: (ESI Mꢁ1) 301. HRMS: Calcd for C19H14N2O2; Exact mass:
302.1055, found (ESI MꢁH) 301.1074.
LRMS: (ESI Mꢁ1) 361. HRMS: Calcd for C18H7F5N2O; Exact
mass: 362.0479, found (ESI MꢁH) 316.0465.
4.2.20. (E)-2-(1H-Indole-3-carbonyl)-3-(naphthalen-2-yl)acrylo
nitrile (22)
4.2.16. (E)-2-(1H-Indole-3-carbonyl)-3-(4-nitrophenyl)acrylo
nitrile (18)
Synthesized using the general procedure as for (14), from 3-
(1H-indol-3-yl)-3-oxopropanenitrile and 2-naphthaldehyde to af-
ford (22) as a yellow solid; 72%; 302–304 °C.
Synthesized using the general procedure as for (14), from 3-
(1H-indol-3-yl)-3-oxopropanenitrile and 4-nitrobenzaldehyde to
afford (18) as a red solid; 75%; 286–288 °C.
1H NMR (DMSO-d6) (300 MHz): d 12.29 (br, NH), 8.54–8.50 (m,
2H, Ind H2 + Ind H4), 8.38 (s, 1H, CH@C), 8.24–8.20 (m, 2H, Ar
H5 + Ar H8), 8.11–7.99 (m, 3H, Ind H7 + Ar H1 + Ar H4), 7.70–
7.54 (m, 3H, Ar H3 + Ar H6 + Ar H7), 7.32–7.24 (m, 2H, Ind
H5 + Ind H6);
1H NMR (DMSO-d6) (300 MHz): d 12.37 (br, NH), 8.50 (s, 1H, Ind
H2), 8.40 (d, J = 8.8 Hz, 2H, Ar H3 + Ar H5), 8.34 (s, 1H, CH@C),
8.23–8.16 (m, 3H, Ar H2 + Ar H6 + Ind H4), 7.56–7.53 (m, 1H, Ind
H7), 7.30–7.27 (m, 2H, Ind H5 + Ind H6);
13C NMR (DMSO-d6) (75 MHz): d 180.7, 149.3, 148.6, 138.6,
136.8, 136.7, 131.0 (2 ꢂ Ar), 125.9, 123.9 (2 ꢂ Ar), 123.7, 122.5,
121.2, 116.7, 115.0, 113.3, 112.5;
13C NMR (DMSO-d6) (75 MHz): d 181.3, 152.0, 136.6, 135.9,
134.3, 132.8, 132.4, 130.0, 128.9, 128.6, 128.5, 127.7, 127.2,
126.1, 124.8, 123.5, 122.4, 121.3, 117.8, 112.5, 112.4, 111.4;
IR (KBr) cmꢁ1: 3174 (NH), 2214 (CN), 1597 (C@O), 1516 (Ar);
LRMS: (ESI Mꢁ1) 321. HRMS: Calcd for C22H14N2O; Exact mass:
322.1106, found (ESI MꢁH) 321.1104.
IR (KBr) cmꢁ1: 3331 (NH), 2219 (CN), 1625 (C@C), 1599 (Ar),
1344 (N–O);
LRMS: (ESI Mꢁ1) 316. HRMS: Calcd for C18H11N3O3; Exact mass:
317.0800, found (ESI MꢁH) 316.0791.
4.2.21. (E)-3-(1H-Indol-3-yl)-2-(1H-indole-3-carbonyl)acryloni
trile (23)
Synthesized using the general procedure as for (14), from 3-
(1H-indol-3-yl)-3-oxopropanenitrile 1H-indole-3-carbaldehyde to
afford (23) as a yellow solid; 79%; 292–294 °C.
4.2.17. (E)-2-(1H-Indole-3-carbonyl)-3-(3,4-dichlorophenyl)
acrylonitrile (19)
Synthesized using the general procedure as for (14), from 3-
(1H-indol-3-yl)-3-oxopropanenitrile and 3,4-dichlorobenzalde-
hyde to afford (19) as a yellow solid; 88%; 288–290 °C.
1H NMR (DMSO-d6) (300 MHz): d 12.34 (br, NH), 8.46 (s 1H, Ar1
H2), 8.24–8.20 (m, 3H, Ar1 H4 + Ar1 H7 + CH@C), 8.05–8.01 (m, 1H,
Ar2 H5), 7.89–8.86 (m, 1H, Ar2 H2), 7.54–7.52 (m, 1H, Ar2 H6),
7.34–7.25 (m, 2H, Ar1 H5 + Ar1 H6);
1H NMR (DMSO-d6) (300 MHz): d 12.14 (br, 2H), 8.63 (s, 1H, Ind
H2), 8.60 (s, 1H, CH@C), 8.47 (s, 1H, Ind2 H2), 8.24–8.22 (m, 1H, Ind
H4), 7.94–7.91 (m, 1H, Ind2 H4), 7.59–7.53 (m, 2H, Ind H7 + Ind2
H7), 7.31–7.21 (m, 4H, Ind H5 + Ind H6 + Ind2 H5 + Ind2 H6);
13C NMR (DMSO-d6) (75 MHz): d 180.7, 145.0, 136.3, 136.1,
133.7, 131.2, 127.2, 126.4, 123.3, 123.1, 121.9, 121.7, 121.5,
120.5, 118.5, 114.3, 112.7, 112.2, 110.3, 101.9;
13C NMR (DMSO-d6) (75 MHz): d 180.8, 149.0, 136.7, 136.6,
134.3, 133.0, 132.0, 131.7, 131.2, 129.4, 125.9, 123.6, 122.5,
121.2, 116.9, 116.8, 113.3, 112.4;
IR (KBr) cmꢁ1: 3262 (NH), 2200 (CN), 1612 (C@O);
LRMS: (ESI Mꢁ1) 310. HRMS: Calcd for C20H13N3O; Exact mass:
311.1059, found (ESI MꢁH) 310.0987.
IR (KBr) cmꢁ1: 3230 (NH), 2223 (CN), 1635 (C@C), 1594 (C@O),
755 (Ar–Cl);
LRMS: (ESI Mꢁ1) 339. HRMS: Calcd for C18H10Cl2N2O; Exact
mass: 340.0170, found (ESI MꢁH) 339.0198.
4.2.22. (E)-2-Cyano-N-(4-methoxybenzyl)-3-(1H-pyrrol-2-yl)
acrylamide (27)
4.2.18. (E)-2-(1H-Indole-3-carbonyl)-3-(4-(trifluoromethyl)
phenyl)acrylonitrile (20)
Methyl 2-cyanoacetate (24) (0.72, 7.28 mmol) was added to a
solution of (4-methoxybenzylamine (1.0 g, 7.28 mmol) in MeOH
(4 mL). The resultant solution was heated under microwave radia-
tion for 15 min at 200 W and 120 °C. After this period, the reaction
was cooled in the freezer for 30 min upon which a crystalline solid
was formed. The solid was collected by filtration, washed with ice
cold MeOH (2 ꢂ 5 mL) and dried under vacuum to afford N-(4-
methoxybenzyl)propionamide (25); 65%. Next, N-(4-methoxyben-
zyl)propionamide (3.77 mmol) was added to an ethanolic solution
(4 mL) of 1H-pyrrole-2-carbaldehyde (4.15 mmol) and piperidine
(2 drops). This mixture was heated under microwave radiation
for 15 min at 200 W and 120 °C. After this period, the solution
was cooled and the solvent removed in vacuo to yield a brown
crude solid, which was purified by flash chromatography (2:8
Synthesized using the general procedure as for (14), from 3-
(1H-indol-3-yl)-3-oxopropanenitrile and 4-(trifluoromethyl)benz-
aldehyde to afford (20) as a yellow solid; 73%; 242–243 °C.
1H NMR (DMSO-d6) (300 MHz): d 12.30 (br, NH), 8.48 (s, 1H, Ind
H2), 8.31 (s, 1H, CH@C), 8.20–8.18 (m, 3H, Ar H2 + Ar H6 + Ind H4),
7.95 (d, J = 7.8 Hz, 2H, Ar H3 + Ar H5), 7.57–7.53 (m, 1H, Ind H7),
7.31–7.26 (m, 2H, Ind H5 + Ind H6);
13C NMR (DMSO-d6) (75 MHz): d 180.8, 150.0, 136.7, 136.5,
136.4, 130.6 (2 ꢂ Ar), 125.9 (2 ꢂ Ar), 125.8, 125.7, 125.6, 123.6,
122.5, 121.2, 116.9, 114.2, 113.4, 112.6;
IR (KBr) cmꢁ1: 3211 (NH), 2223 (CN), 1596 (C@O), 1336 (C–F);
LRMS: (ESI Mꢁ1) 339. HRMS: Calcd for C19H11F3N2O; Exact
mass: 340.0823, found ESI (MꢁH) 339.0829.

344
M. Tarleton et al. / Bioorg. Med. Chem. 21 (2013) 333–347
EtOAc/Hexanes) to afford (27) as a brown solid; 81%; mp 203–
204 °C.
4.2.26. (E)-2-Cyano-N-(4-methoxybenzyl)-3-(5-methylfuran-2-
yl)acrylamide (31)
1H NMR (Acetone-d6) (300 MHz): d 10.94 (br, NH), 8.18 (s, 1H,
CH@C), 7.65 (br, NH), 7.33–7.28 (m, 4H, Ar H2 + Ar H6 + Pyr
H3 + Pyr H5), 6.87 (d, J = 8.7 Hz, 2H, Ar H3 + Ar H5), 6.43 (m, 1H,
Pyr H4), 4.49 (d, J = 6.0 Hz, 2H, CH2NH), 3.77 (s, 3H, OCH3);
13C NMR (Acetone-d6) (75 MHz): d 160.7, 158.4, 139.6, 131.3,
130.7, 128.4 (2 ꢂ Ar), 125.7, 117.1, 116.5, 113.1 (2 ꢂ Ar), 111.6,
94.8, 54.0, 42.4;
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and 5-methylfuran-2-carbaldehyde
to afford (31) as a white solid; 92%; 146–147 °C.
1H NMR (CDCl3) (300 MHz): d 7.94 (s, 1H, CH@O), 7.83 (br, NH),
7.32–7.26 (m, 3H, Ar H2 + Ar H6 + Fur H3), 6.88 (d, J = 8.7 Hz, 2H,
Ar H3 + Ar H5), 6.43–6.42 (m, 1H), Fur H4), 4.49 (d, J = 5.9 Hz, 2H,
CH2NH), 3.77 (s, 3H, OCH3), 2.42 (s, 3H, CH3);
IR (KBr) cmꢁ1: 3360 (NH), 3234 (NH), 2202 (CN), 1650 (C@O);
LRMS: (ESI M+1) 282. HRMS: Calcd for C16H15N3O2; Exact mass:
281.1164, found (ESI M+H) 282.1208.
13C NMR (CDCl3) (75 MHz): d 160.0, 158.4, 147.3, 135.4, 130.6,
128.5 (2 ꢂ Ar), 122.3, 115.6, 113.1 (2 ꢂ Ar), 110.0, 99.9, 98.3,
54.0, 42.5, 12.5;
IR (KBr) cmꢁ1: 3364 (NH), 2214 (CN), 1671 (C@O);
LRMS: (ESI M+1) 297. HRMS: Calcd for C17H16N2O3; Exact mass:
296.1161, found (ESI M+H) 297.1206.
4.2.23. (E)-2-Cyano-N-(3,4-dichlorobenzyl)-3-(1H-pyrrol-2-yl)
acrylamide (28)
Synthesized using the general procedure as for (27), from N-
(3,4-dichlorobenzyl)propioamide and 1H-pyrrole-2-carbaldehyde
to afford (28) as a yellow solid; 80%; 234–235 °C.
1H NMR (DMSO-d6) (300 MHz): d 11.88 (br, NH), 8.67 (t,
J = 5.9 Hz, NH), 8.05 (s, 1H, CH@C), 7.58–7.53 (m, 2H, Ar H5 + Ar
H2), 7.30–7.26 (m, 3H, Ar H6 + Pyr H5 + Pyr H3), 6.42–6.40 (m,
1H, Pyr H4), 4.36 (d, J = 5.9 Hz, 2H, CH2NH);
4.2.27. (E)-2-Cyano-3-(5-(hydroxymethyl)furan-2-yl)-N-(4-
methoxybenzyl)acrylamide (32)
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and 5-(hydroxymethyl)furan-2-carb-
aldehyde to afford (32) as a white solid; 45%; 130–132 °C.
1H NMR (Acetone-d6) (300 MHz): d 7.98 (s, 1H, CH@O), 7.82 (br,
NH), 7.33–7.29 (m, 3H, Ar H2 + Ar H6 + Fur H3), 6.88 (d, J = 8.7 Hz,
2H, Ar H3 + Ar H5), 6.62 (d, J = 3.5 Hz, 1H, Fur H4), 4.65 (s, 2H,
CH2OH), 4.50 (br, OH), 4.49 (d, J = 5.9 Hz, 2H, CH2NH), 3.77 (s,
3H, OCH3);
13C NMR (DMSO-d6) (75 MHz): d 161.8, 141.1, 140.5, 140.3,
130.7, 130.4, 129.3, 127.7, 126.5, 126.3, 117.7, 115.5, 112.5, 94.3,
42.0;
IR (KBr) cmꢁ1: 3342 (NH), 3232 (NH), 2209 (CN), 1643 (C@O),
749 (Ar–Cl);
13C NMR (Acetone-d6) (75 MHz): d 161.1, 159.8, 158.5, 147.8,
135.7, 130.5, 128.5 (2 ꢂ Ar), 121.1, 115.4, 113.2 (2 ꢂ Ar), 109.9,
99.6, 56.1, 54.1, 42.6;
LRMS: (ESI Mꢁ1) 318. HRMS: Calcd for C15H11Cl2N3O; Exact
IR (KBr) cmꢁ1: 3347 (NH), 3235 (OH), 2215 (CN), 1667 (C@O),
1248 (C–O);
mass: 319.0279, found (ESI MꢁH) 318.0198.
LRMS: (ESI M+1) 313. HRMS: Calcd for C17H16N2O4; Exact mass:
312.1110, found (ESI M+H) 313.1122.
4.2.24. (E)-2-Cyano-3-(furan-2-yl)-N-(4-methoxybenzyl)acryl
amide (29)
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and furan-2-carbaldehyde to afford
(29) as a orange solid; 67%; 119–120 °C.
4.2.28. (E)-3-(5-Chlorofuran-2-yl)-2-cyano-N-(4-methoxyben
zyl)acrylamide (33)
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and 5-chlorofuran-2-carbaldehyde to
afford (33) as a yellow solid; 71%; 164–165 °C.
1H NMR (Acetone-d6) (300 MHz): d 8.03 (s, 1H, CH@C), 7.94 (d,
J = 1.6 Hz, 1H, Fur H5), 7.86 (br, NH), 7.36 (d, J = 3.6 Hz, 1H, Fur H3),
7.30 (d, J = 8.6 Hz, 2H, Ar H2 + Ar H6), 6.88 (d, J = 8.6 Hz, 2H, Ar
H3 + Ar H5), 6.77 (dd, J = 1.6 Hz, 3.6 Hz, 1H, Fur H4), 4.50 (d,
J = 5.9 Hz, 2H, CH2NH), 3.77 (s, 3H, OCH3);
1H NMR (CDCl3) (300 MHz): d 7.99 (s, 1H, CH@C), 7.25 (d,
J = 8.5 Hz, 2H, Ar H2 + Ar H6), 7.20–7.19 (m, 1H, Fur H3), 6.88 (d,
J = 8.5 Hz, 2H, Ar H3 + Ar H6), 6.55 (br, NH), 6.44–6.43 (m, 1H,
Fur H4), 4.51 (d, J = 5.6 Hz, 2H, CH2NH), 3.81 (s, 3H, OCH3);
13C NMR (CDCl3) (75 MHz): d 159.3, 158.8, 147.9, 142.5, 135.6,
128.8 (2 ꢂ Ar), 128.6, 122.0, 115.8, 113.7 (2 ꢂ Ar), 110.1, 99.4,
54.8, 43.5;
13C NMR (Acetone-d6) (75 MHz): d 159.7, 158.5, 148.6, 147.4,
135.7, 130.4, 128.5 (2 ꢂ Ar), 120.1, 115.3, 113.2 (2 ꢂ Ar), 113.0,
100.6, 54.1, 42.4;
IR (KBr) cmꢁ1: 3325 (NH), 2225 (CN), 1659 (C@O);
LRMS: (ESI M+1) 283. HRMS: Calcd for C16H14N2O3; Exact mass
282.1004, found (ESI M+H) 283.0989.
IR (KBr) cmꢁ1: 3342 (NH), 2217 (CN), 1666 (C@O), 1251 (C–O);
LRMS: (ESI M+1) 317. HRMS: Calcd for C16H13ClN2O3; Exact
mass: 316.0615, found (ESI M+H) 317.0666.
4.2.25. (E)-2-Cyano-N-(3,4-dichlorobenzyl)-3-(furan-2-yl)acryl
amide (30)
4.2.29. (E)-3-(5-Bromofuran-2-yl)-2-cyano-N-(4-methoxyben
zyl)acrylamide (34)
Synthesized using the general procedure as for (27), from N-
(3,4-dichlorobenzyl)propioamide and furan-2-carbaldehyde to af-
ford (30) as a light yellow solid; 76%; 198–199 °C.
1H NMR (DMSO-d6) (300 MHz): d 8.92 (t, J = 5.9 Hz, NH), 8.12 (d,
J = 1.6 Hz, 1H, Fur H5), 7.99 (s, 1H, CH@C), 7.58 (d, J = 8.3 Hz, 1H, Ar
H5), 7.55 (d, J = 2.0 Hz, 1H, Ar H1), 7.37 (d, J = 3.6 Hz, 1H, Fur H3),
7.29 (dd, J = 2.0, 8.3 Hz, 1H, Ar H6), 6.81 (dd, J = 1.6, 3.6 Hz, 1H,
Fur H4), 4.37 (d, J = 5.9 Hz, 2H, CH2NH);
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and 5-bromofuran-2-carbaldehyde
to afford (34) as a yellow solid; 84%; 160–162 °C.
1H NMR (CDCl3) (300 MHz): d 8.01 (s, 1H, CH@C), 7.25 (d,
J = 8.6 Hz, 2H, Ar H2 + Ar H6), 7.16 (d, J = 3.7 Hz, 1H, Fur H3), 6.89
(d, J = 8.6 Hz, 2H, Ar H3 + Ar H5), 6.58-6.57 (m, 2H, Fur H4 + NH),
4.51 (d, J = 5.7 Hz, 2H, CH2NH), 3.80 (s, 3H, OCH3);
13C NMR (CDCl3) (75 MHz): d 159.3, 158.7, 150.2, 135.5, 129.1,
128.8 (2 ꢂ Ar), 128.6, 122.0, 115.7, 115.1, 113.7 (2 ꢂ Ar), 99.5,
54.8, 43.5;
13C NMR (DMSO-d6) (75 MHz): d 161.0, 148.7, 148.3, 140.1,
136.1, 130.7, 130.4, 129.4, 127.7, 121.8, 116.0, 113.8, 103.4,
100.3, 42.1;
IR (KBr) cmꢁ1: 3348 (NH), 2219 (CN), 1672 (C@O), 1247 (C–O),
807 (Ar–Br);
LRMS: (ESI M+1) 361. HRMS: Calcd for: C16H13BrN2O3; Exact
mass: 360.0110, found (ESI M+H) 361.0145.
IR (KBr) cmꢁ1: 3358 (NH), 2216 (CN), 1671 (C@O), 757 (Ar–Cl);
LRMS: (ESI Mꢁ1) 319. HRMS: Calcd for C15H10Cl2N2O2; Exact
mass: 320.0119, found (ESI MꢁH) 319.0039.

M. Tarleton et al. / Bioorg. Med. Chem. 21 (2013) 333–347
345
4.2.30. (E)-2-Cyano-N-(4-methoxybenzyl)-3-(5-phenylfuran-2-
yl)acrylamide (35)
IR (KBr) cmꢁ1: 3342 (NH), 2225 (CN), 1672 (C@O), 1513 (Ar),
1251 (C–O);
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and 5-phenylfuran-2-carbaldehyde
to afford (35) as a yellow solid; 85%; 203–205 °C.
1H NMR (DMSO-d6) (300 MHz): d 8.81 (t, J = 5.9 Hz, NH), 7.99 (s,
CH@C), 7.90 (d, J = 7.3 Hz, 2H, Ar2 H2 + Ar2 H6), 7.53–7.48 (m, 2H,
Ar2 H3 + Ar2 H5), 7.44–7.42 (m, 2H, Ar2 H4 + Fur H3), 7.34–7.33
(m, 1H, Fur H4), 7.23 (d, J = 8.6 Hz, 2H, Ar H2 + Ar H6), 6.88 (d
J = 8.6 Hz, 2H, Ar H3 + Ar H5), 4.32 (d, J = 5.9 Hz, 2H, CH2NH),
3.71 (s, 3H, OCH3);
LRMS: (ESI M+1) 333. HRMS: Calcd for C20H16N2O3; Exact mass:
322.1161, found (ESI M+H) 333.1231.
4.2.34. (E)-2-Cyano-N-(4-methoxybenzyl)-3-phenylacrylamide
(39)
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and benzaldehyde to afford (39) as a
white solid; 63%; 117–118 °C.
1H NMR (Acetone-d6) (300 MHz): d 8.27 (s, 1H, CH@C), 8.00 (m,
32H, Ar2 H2 + Ar2 H6, NH), 7.57 (m, 3H, Ar2 H3 + Ar2 H4 + Ar2 H5),
7.32 (d, J = 8.7 Hz, 2H, Ar H2 + Ar H6), 6.89 (d, J = 8.7 Hz, 2H, Ar
H3 + Ar H5), 4.52 (d, J = 5.9 Hz, 2H, CH2NH), 3.78 (s, 3H, OCH3);
13C NMR (Acetone-d6) (75 MHz): d 159.7, 158.5, 150.7, 131.8
(2 ꢂ Ar), 130.9, 130.3, 129.7 (2 ꢂ Ar), 128.6 (2 ꢂ Ar), 115.6, 113.2
(2 ꢂ Ar), 105.4, 104.4, 54.0, 42.6;
13C NMR (DMSO-d6) (75 MHz): d 160.9, 158.2, 157.7, 147.7,
135.1, 130.9, 129.5, 129.1 (2 ꢂ Ar), 128.7 (2 ꢂ Ar), 128.6, 124.7
(2 ꢂ Ar), 124.6, 116.6, 113.6 (2 ꢂ Ar), 109.6, 99.7, 54.9, 42.6;
IR (KBr) cmꢁ1: 3362 (NH), 2209 (CN), 1666 (C@O), 1607 (Ar),
1247 (C–O);
LRMS: (ESI M+1) 359. HRMS: Calcd for C22H18N2O3; Exact mass:
358.1317, found (ESI M+H) 359.1489.
IR (KBr) cmꢁ1: 3336 (NH), 2223 (CN), 1663 (C@O), 1259 (C–O);
LRMS: (ESI M+1) 293. HRMS: Calcd for C18H16N2O2; Exact mass:
292.1212, found (ESI M+H) 293.1215.
4.2.31. (E)-3-(4-Bromofuran-2-yl)-2-cyano-N-(4-methoxyben
zyl)acrylamide (36)
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and 4-bromofuran-2-carbaldehyde
to afford (36) as a white solid; 70%; 169–170 °C.
1H NMR (CDCl3) (300 MHz): d 8.04 (s, 1H, CH@C), 7.68 (s, 1H,
Fur H5), 7.26–7.23 (m, 3H, Ar H2 + Ar H6 + Fur H3), 6.88 (d,
J = 8.6 Hz, 2H, Ar H3 + Ar H5), 6.59 (br, NH), 4.51 (d, J = 5.7 Hz,
2H, CH2NH), 3.80 (s, 3H, OCH3);
4.2.35. (E)-2-Cyano-N-(4-methoxybenzyl)-3-p-tolylacrylamide
(40)
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and 4-methylbenzaldehyde to afford
(40) as a yellow solid; 70%; 115–117 °C.
1H NMR (Acetone-d6) (300 MHz): d 8.22 (s, 1H, CH@C), 7.90 (d,
J = 8.2 Hz, 2H, Ar2 H2 + Ar 2 H6), 7.38 (d, J = 8.2 Hz, 2H, Ar 2
H3 + Ar2 H5), 7.32 (d, J = 8.8 Hz, 2H, Ar H2 + Ar H6), 6.88 (d,
J = 8.8 Hz, 2H, Ar H3 + Ar H5), 4.51 (d, J = 5.9 Hz, 2H, CH2NH),
3.77 (s, 3H, OCH3), 2.41 (s, 3H, CH3);
13C NMR (CDCl3) (75 MHz): d 159.0, 158.8, 148.9, 144.8, 136.1,
128.8 (2 ꢂ Ar), 128.5, 121.7, 115.6, 113.7 (2 ꢂ Ar), 102.6, 101.1,
54.8, 43.6;
IR (KBr) cmꢁ1: 3332 (NH), 2223 (CN), 1660 (C@O), 1254 (C–O),
926 (Ar–Br);
LRMS: (ESI M+1) 361. HRMS: Calcd for C16H13BrN2O3; Exact
mass: 360.0110, found (ESI M+H) 361.0195.
13C NMR (Acetone-d6) (75 MHz): d 159.9, 158.4, 150.7, 142.8,
130.4, 129.9 (2 ꢂ Ar), 129.3 (2 ꢂ Ar), 129.1, 128.6 (2 ꢂ Ar), 115.7,
113.2 (2 ꢂ Ar), 103.9, 54.0, 42.6, 20.2;
IR (KBr) cmꢁ1: 3339 (NH), 2220 (CN), 1662 (C@O), 1259 (C–O);
LRMS: (ESI M+1) 307. HRMS: Calcd for C19H18N2O2; Exact mass:
306.1368, found (ESI M+H) 307.1435.
4.2.32. (E)-2-Cyano-3-(furan-3-yl)-N-(4-methoxybenzyl)acryl
amide (37)
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and furan-3-carbaldehyde to afford
(37) as a brown solid; 60%; 122–124 °C.
4.2.36. (E)-3-(4-Chlorophenyl)-2-cyano-N-(4-methoxybenzyl)
acrylamide (41)
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and 4-chlorobenzaldehyde to afford
(41) as a yellow solid; 28%; 134–135 °C.
1H NMR (Acetone-d6) (300 MHz): d 8.32 (s, 1H, Fur H2), 8.21 (s,
1H, CH@C), 7.84 (br, NH), 7.78-7.77 (m, 1H, Fur H5), 7.30 (d
J = 8.7 Hz, 2H, Ar H2 + Ar H6), 7.23–7.22 (m, 1H, Fur H4), 6.88 (d,
J = 8.7 Hz, 2H, Ar H3 + Ar H5), 4.49 (d, J = 6.0 Hz, 2H, CH2NH),
3.77 (s, 3H, OCH3);
1H NMR (Acetone-d6) (300 MHz): d 8.25 (s, 1H, CH@C), 8.01 (d,
J = 8.8 Hz, 2H, Ar2 H2 + Ar2 H6), 7.60 (d, J = 8.8 Hz, 2H, Ar2
H3 + Ar2 H5), 7.31 (d, J = 8.7 Hz, 2H, Ar H2 + Ar H6), 6.89 (d,
J = 8.7 Hz, 2H, Ar H3 + Ar H5), 4.51 (d, J = 5.9 Hz, 2H, CH2NH),
3.77 (s, 3H, OCH3);
13C NMR (Acetone-d6) (75 MHz): d 159.7, 158.5, 149.4, 145.0,
141.6, 130.4, 128.5 (2 ꢂ Ar), 120.4, 115.8, 113.2, (2 ꢂ Ar), 107.3,
103.8, 54.1, 42.5;
13C NMR (Acetone-d6) (75 MHz): d 159.5, 158.5, 149.3, 137.1,
131.3 (2 ꢂ Ar), 130.5, 130.3, 128.8 (2 ꢂ Ar), 128.6 (2 ꢂ Ar), 115.4,
113.2 (2 ꢂ Ar), 106.0, 54.1, 42.6;
IR (KBr) cmꢁ1: 3332 (NH), 2213 (CN), 1661 (C@O), 1611 (Ar),
1513 (Ar), 1251 (C–O). LRMS: (ESI M+1) 283;
HRMS: Calcd for Chemical Formula: C16H14N2O3; Exact mass:
282.1004, found (ESI M+H) 283.1032.
IR (KBr) cmꢁ1: 3361 (NH), 2216 (CN), 1664 (C@O), 1252 (C–O),
819 (Ar–Cl);
LRMS: (ESI M+1) 327. HRMS: Calcd for C18H15ClN2O2; Exact
mass: 326.0822, found (ESI M+H) 327.0846.
4.2.33. (E)-3-(Benzofuran-2-yl)-2-cyano-N-(4-methoxybenzyl)
acrylamide (38)
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and benzofuran-2-carbaldehyde to
afford (38) as a white solid; 87%; 150–153 °C.
4.2.37. (E)-2-Cyano-3-(4-hydroxyphenyl)-N-(4-methoxybenzyl)
acrylamide (42)
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and 4-hydroxybenzaldehyde to af-
ford (42) as a light yellow solid; 20%; 197–198 °C.
1H NMR (Acetone-d6) (300 MHz): d 8.16 (s, 1H, CH@C), 7.94 (d,
J = 8.8 Hz, 2H, Ar H3 + Ar H5), 7.81 (br, NH), 7.31 (d, J = 8.6 Hz, 2H,
Ar2 H2 + Ar2 H6), 7.00 (d, J = 8.8 Hz, 2H, Ar H2 + Ar H6), 6.88 (d,
J = 8.6 Hz, 2H, Ar2 H3 + Ar2 H5), 4.50 (d, J = 5.9 Hz, 2H, CH2NH),
3.77 (s, 3H, OCH3);
1H NMR (DMSO-d6) (300 MHz): d 8.96 (t, J = 5.8 Hz, NH), 8.14 (s,
1H, CH@C), 7.83–7.80 (m, 1H, Ar2 H4), 7.76 (s, 1H, Ar2 H3), 7.68–
7.65 (m, 1H, Ar2 H7), 7.54–7.49 (m, 1H, Ar2 H6), 7.38–7.32 (m, 1H,
Ar2 H5), 7.25 (d, J = 8.5 Hz, 2H, Ar H2 + Ar H6), 6.88 (d, J = 8.5 Hz,
2H, Ar H3 + Ar H5), 4.34 (d, J = 5.8 Hz, 2H, CH2NH), 3.72 (s, 3H, OCH3);
13C NMR (DMSO-d6) (75 MHz): d 160.4, 158.2, 155.3, 149.6,
136.3, 130.7, 128.8 (2 ꢂ Ar), 128.5, 127.3, 124.1, 123.0, 117.3,
115.7, 113.6 (2 ꢂ Ar), 111.6, 104.1, 55.0, 42.6;

346
M. Tarleton et al. / Bioorg. Med. Chem. 21 (2013) 333–347
13C NMR (Acetone-d6) (75 MHz): d 161.0, 160.2, 150.5, 132.5
1H NMR (Acetone-d6) (300 MHz): d 8.29 (s, 1H, CH@C), 8.22 (br,
NH), 8.02–7.99 (m, 2H, Ar2 H2 + Ar2 H6), 7.61–7.51 (m, 5H, Ar2
H3 + Ar2 H4 + Ar2 H5 + Ar H1 + Ar H5), 7.38 (dd, J = 2.0, 8.3 Hz,
1H, Ar H6), 4.60 (d, J = 6.0 Hz, 2H, CH2NH);
(2 ꢂ Ar), 130.6, 128.5 (2 ꢂ Ar), 123.4, 116.4, 115.6 (2 ꢂ Ar), 113.1
(2 ꢂ Ar), 102.8, 100.8, 54.0, 42.5;
IR (KBr) cmꢁ1: 3342 (NH), 3153 (OH), 2212 (CN), 1645 (C@O),
1172 (C–O);
13C NMR (Acetone-d6) (75 MHz): d 160.2, 151.2, 139.6, 131.9,
131.7, 131.1, 129.9, 129.8 (2 ꢂ Ar), 129.5, 129.3 (2 ꢂ Ar), 128.6,
127.3, 115.5, 104.9, 42.1;
LRMS: (ESI M+1) 309. HRMS: Calcd for C18H16N2O3; Exact mass:
308.3312, found (ESI M+H) 309.1234.
IR (KBr) cmꢁ1: 3364 (NH), 2215 (CN), 1678 (C@O), 679 (Ar–Cl);
LRMS: (ESI Mꢁ1) 329. HRMS: Calcd for C17H12Cl2N2O; Exact
mass: 330.0327, found (ESI MꢁH) 329.0273.
4.2.38. (E)-2-Cyano-N-(4-methoxyphenyl)-3-(4-methoxybenzyl)
acrylamide (43)
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and 4-methoxybenzaldehyde to af-
ford (43) as a yellow solid; 34%; 138–139 °C.
4.2.42. (E)-2-Cyano-N-(3,4-dichlorobenzyl)-3-p-tolylacrylamide
(47)
1H NMR (Acetone-d6) (300 MHz): d 8.19 (s, 1H, CH@C), 8.01 (d,
J = 8.9 Hz, 2H, Ar2 H2 + Ar2 H6), 7.84 (br, NH), 7.31 (d, J = 8.5 Hz,
2H, Ar H2 + Ar H6), 7.10 (d, J = 8.9 Hz, 2H, Ar2 H3 + Ar2 H5), 6.89
(d, J = 8.5 Hz, 2H, Ar H3 + Ar H5), 4.50 (d, J = 6.0 Hz, 2H, CH2NH),
3.91 (s, 3H, OCH3), 3.77 (s, 3H, OCH3);
Synthesized using the general procedure as for (27), from N-
(3,4-dichlorobenzyl)propioamide and 4-methylbenzaldehyde to
afford (47) as a light yellow solid; 62%; 161–162 °C.
1H NMR (DMSO-d6) (300 MHz): d 8.99 (t, J = 5.8 Hz, NH), 8.16 (s,
1H, CH@C), 7.85 (d, J = 8.1 Hz, 2H. Ar2 H2 + Ar2 H6), 7.59–7.56 (m,
2H, Ar H5 + Ar H1), 7.36 (d, J = 8.1 Hz, 2H, Ar2 H3 + Ar2 H6), 7.31
(dd, J = 1.9, 8.3 Hz, 1H, Ar H6), 4.40 (d, J = 5.8 Hz, 2H, CH2NH),
2.37 (s, 3H, OCH3);
13C NMR (Acetone-d6) (75 MHz): d 162.7, 160.2, 158.5, 150.3,
132.1 (2 ꢂ Ar), 130.5, 128.5 (2 ꢂ Ar), 124.3, 116.5, 114.1 (2 ꢂ Ar),
113.1 (2 ꢂ Ar), 101.6, 54.6, 54.0, 42.5;
IR (KBr) cmꢁ1: 3420 (NH), 2200 (CN), 1667 (C@O), 1250 (C–O),
1178 (C-O);
13C NMR (DMSO-d6) (75 MHz): d 161.2, 150.9, 143.0, 140.1,
130.8, 130.4, 130.1, 129.8 (2 ꢂ Ar), 129.7 (2 ꢂ Ar), 129.4, 129.1,
127.8, 116.4, 104.5, 42.1, 21.1;
LRMS: (ESI M+1) 323. HRMS: Calcd for C19H18N2O3; Exact mass:
322.1317, found (ESI M+H) 323.1378.
IR (KBr) cmꢁ1: 3366 (NH), 2217 (CN), 1681 (C@O), 813 (Ar–Cl);
LRMS: (ESI Mꢁ1) 343. HRMS: Calcd for C18H14Cl2N2O; Exact
mass: 344.0483, found (ESI MꢁH) 343.0446.
4.2.39. (E)-2-Cyano-N-(4-methoxybenzyl)-3-(naphthalen-2-yl)
acrylamide (44)
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and 2-naphthaldehyde to afford (44)
as a yellow solid; 45%; 143–144 °C.
4.2.43. (E)-3-(4-Chlorophenyl)-2-cyano-N-(3,4-
dichlorobenzyl)acrylamide (48)
Synthesized using the general procedure as for (27), from N-
(3,4-dichlorobenzyl)propioamide and 4-chlorobenzaldehyde to af-
ford (48) as a yellow solid; 30%; 156–158 °C.
1H NMR (Acetone-d6) (300 MHz): d 8.49 (s, 1H CH@C), 8.43 (s,
1H, Ar2 H1), 8.20 (dd, J = 1.7, 8.7 Hz, 1H, Ar2 H8), 8.09–7.99 (m,
4H, Ar2 H5 + Ar2 H4 + Ar2 H6 + NH), 7.70–7.60 (m, 2H, Ar2
H5 + Ar2 H3), 7.34 (d, J = 8.7 Hz, 2H, Ar H2 + Ar H6), 6.90 (d,
J = 8.7 Hz, 2H, Ar H3 + Ar H5), 4.54 (d, J = 5.9 Hz, 2H, CH2NH),
3.78 (s, 3H, OCH3);
1H NMR (Acetone-d6) (300 MHz): d 8.27 (s, 1H, CH@C), 8.24 (br,
NH), 8.02 (d, J = 8.5 Hz, 2H, Ar2 H2 + Ar2 H6), 7.63–7.60 (m, 3H, Ar2
H3 + Ar2 H5 + Ar H5), 7.53 (d, J = 8.3 Hz, 1H, Ar H1), 7.38 (dd,
J = 2.0, 8.3 Hz, 1H, Ar H6), 4.60 (d, J = 6.1 Hz, 2H, CH2NH);
13C NMR (Acetone-d6) (75 MHz): d 159.9, 149.7, 139.5, 138.3,
137.2, 131.3 (2 ꢂ Ar), 131.1, 130.5, 129.9, 129.3, 128.8, 127.3
(2 ꢂ Ar), 124.6, 115.3, 42.2;
13C NMR (DMSO-d6) (75 MHz): d 160.9, 158.2, 150.5, 134.2,
132.6, 132.3, 130.7, 129.4, 128.9 (2 ꢂ Ar), 128.8, 128.7, 128.6,
127.7, 127.2, 124.5, 116.4, 113.6 (2 ꢂ Ar), 106.1, 55.0, 42.6;
IR (KBr) cmꢁ1: 3371 (NH), 2212 (CN), 1676 (C@O), 1253 (C–O);
LRMS: (ESI M+1) 343. HRMS: Calcd for C22H18N2O2; Exact mass:
342.1368, found (ESI M+H) 343.1527.
IR (KBr) cmꢁ1: 3384 (NH), 2212 (CN), 1673 (C@O);
LRMS: (ESI Mꢁ1) 363. HRMS: Calcd for C17H11Cl3N2O; Exact
mass: 363.9937, found (ESI MꢁH) 363.9459.
4.2.44. (E)-2-Cyano-N-(3,4-dichlorobenzyl)-3-(4-
hydroxyphenyl)acrylamide (49)
4.2.40. (E)-2-Cyano-N-(4-methoxybenzyl)-3-(naphthalen-1-yl)
acrylamide (45)
Synthesized using the general procedure as for (27), from N-(4-
methoxybenzyl)propioamide and 1-naphthaldehyde to afford (45)
as a clear oil; 70%.
Synthesized using the general procedure as for (27), from N-
(3,4-dichlorobenzyl)propioamide and 4-hydroxybenzaldehyde to
afford (49) as a yellow solid; 61%; 230–232 °C.
1H NMR (DMSO-d6) (300 MHz): d 10.57 (br, OH), 8.86 (t,
J = 5.8 Hz, NH), 8.08 (s, 1H, CH@C), 7.87 (d, J = 8.7 Hz, 2H, Ar2
H2 + Ar2 H6), 7.58–7.54 (m, 2H, Ar H1 + Ar H5), 7.29 (dd, J = 2.0,
8.3 Hz, 1H, Ar H6), 6.92 (d, J = 8.7 Hz, 2H, Ar H3 + Ar H5), 4.38 (d,
J = 5.8 Hz, 2H, CH2NH);
1H NMR (Acetone-d6) (300 MHz): d 9.03 (s, 1H, CH@C), 8.14–
8.07 (m, 4H, Ar2 H5 + Ar2 H8 + Ar2 H4 + NH), 8.02–7.99 (m, 1H,
Ar2 H2), 7.67–7.58 (m, 3H, Ar2 H3 + Ar H6 + Ar2 H7), 7.37 (d,
J = 8.7 Hz, 2H, Ar H2 + Ar H6), 6.90 (d, J = 8.7 Hz, 2H, Ar H3 + Ar
H5), 4.57 (d, J = 6.0 Hz, 2H, CH2NH), 3.77 (s, 3H, OCH3);
13C NMR (Acetone-d6) (75 MHz): d 159.7, 158.6, 148.7, 133.1,
131.6, 130.9, 130.3, 128.7 (2 ꢂ Ar), 128.4, 127.0, 126.7, 126.2,
125.3, 124.8, 122.8, 115.3, 113.2 (2 ꢂ Ar), 109.3, 54.1, 42.7;
IR (KBr) cmꢁ1: 3357 (NH), 2227 (CN), 1676 (C@O), 1237 (C–O);
LRMS: (ESI M+1) 343. HRMS: Calcd for C22H18N2O2; Exact mass:
342.1368, found (ESI M+H) 343.1448.
13C NMR (DMSO-d6) (75 MHz): d 161.8, 161.7, 150.8, 140.3,
132.9 (2 ꢂ Ar), 130.7, 130.4, 129.4, 129.3, 127.7, 122.8, 117.0,
116.1 (2 ꢂ Ar), 100.6, 42.1;
IR (KBr) cmꢁ1: 3350 (NH), 3140 (OH), 2217 (CN), 1642 (C@O),
837 (Ar–Cl);
LRMS: (ESI Mꢁ1) 345. HRMS: Calcd for C17H12Cl2N2O2; Exact
mass: 346.0276, found (ESI MꢁH) 345.0249.
4.2.45. (E)-2-Cyano-N-(3,4-dichlorobenzyl)-3-(4-
methoxyphenyl)acrylamide (50)
4.2.41. (E)-2-Cyano-N-(3,4-dichlorobenzyl)-3-phenylacrylamide
(46)
Synthesized using the general procedure as for (27), from N-
(3,4-dichlorobenzyl)propioamide and 4-methoxybenzaldehyde to
afford (50) as a light yellow solid; 58%; 169–170 °C.
Synthesized using the general procedure as for (27), from N-
(3,4-dichlorobenzyl)propioamide and benzaldehyde to afford (46)
as a yellow solid; 57%; 159–160 °C.

M. Tarleton et al. / Bioorg. Med. Chem. 21 (2013) 333–347
347
1H NMR (DMSO-d6) (300 MHz): d 8.91 (t, J = 5.8 Hz, 1H, NH),
Supplementary data
8.14 (s, 1H, CH@C), 7.96 (d, J = 8.9 Hz, 2H, Ar2 H2 + Ar2 H6),
7.59–7.56 (m, 2H, Ar H1 + Ar H5), 7.30 (dd, J = 1.9, 8.2 Hz, 1H, Ar
H6), 7.11 (d, J = 8.9 Hz, 2H, Ar2 H3 + Ar2 H5), 4.39 (d, J = 5.8 Hz,
2H, CH2NH), 3.84 (s, 3H, OCH3);
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.bmc.2012.10.003.
13C NMR (DMSO-d6) (75 MHz): d 162.6, 161.5, 150.5, 140.2,
132.5 (2 ꢂ Ar), 130.7, 130.4, 129.4, 129.4, 127.7, 124.3, 116.8,
114.7 (2 ꢂ Ar), 102.1, 55.5, 42.1;
References and notes
1. www.cancer.org.au/Newsmedia/factsfigures.htm accessed 21/9/2011.
2. McCluskey, A.; Bowyer, M. C.; Collins, E.; Sim, A. T. R.; Sakoff, J. A.; Baldwin, M.
L. Bioorg. Med. Chem. Lett. 2000, 10, 1687.
3. McCluskey, A.; Ackland, S. P.; Gardiner, E.; Baldwin, M. L.; Bowyer, M. C.; Keane,
M. A.; Walkom, C.; Sakoff, J. A. Anti-Cancer Drug Des. 2001, 16, 291.
4. Sakoff, J. A.; Ackland, S. P.; Baldwin, M. L.; Keane, M. A.; McCluskey, A. Invest.
New Drugs 2002, 20, 1.
5. McCluskey, A.; Ackland, S. P.; Bowyer, M. C.; Baldwin, M. L.; Garner, J.; Walkom,
C. C.; Sakoff, J. A. Bioorg. Chem. 2003, 31, 68.
6. Sakoff, J. A.; Howitt, I. J.; Ackland, S. P.; McCluskey, A. Cancer Chemother.
Pharmacol. 2004, 53, 225.
7. Hart, M. E.; Chamberlin, A. R.; Walkom, C.; Sakoff, J. A.; McCluskey, A. Bioorg.
Med. Chem. Lett. 2004, 14, 1969.
8. Hill, T. A.; Stewart, S. G.; Sauer, B.; Gilbert, J.; Ackland, S. P.; Sakoff, J. A.;
McCluskey, A. Bioorg. Med. Chem. Lett. 2007, 17, 3392.
9. Hill, T. A.; Stewart, S. G.; Gordon, C. P.; Ackland, S. P.; Gilbert, J.; Sauer, B.;
Sakoff, J. A.; McCluskey, A. ChemMedChem 2008, 3, 1878.
10. Joshi, S.; Gaddipati, S.; Gilbert, J.; Smith, C. M.; Gordon, C. P.; Sakoff, J. A.;
McCluskey, A.; Robinson, P. J.; Braithwaite, A. W.; Chircop, M. Mol. Cancer Ther.
2010, 9, 1995.
11. Chircop, M.; Malladi, C. S.; Lian, A. T.; Zavortink, M.; Gordon, C. P.; McCluskey,
A.; Robinson, P. J. Cell. Mol. Life Sci. 2010, 21, 3725.
12. Chircop, C.; Perera, S.; Mariana, A.; Lau, H.; Ma, M. P. C.; Gilbert, J.; Jones, N. C.;
Gordon, C. P.; Young, K. A.; Morokoff, A.; Sakoff, J. A.; O’Brien, T. J.; McCluskey,
A.; Robinson, P. J. Mol. Cancer Ther. 2011, 10, 1553.
13. Tarleton, M.; Robertson, M. J.; Gilbert, J.; McCluskey, A.; Sakoff, J. A.
MedChemCommun 2011, 2, 31.
14. Carta, A.; Sanna, P.; Palomba, M.; Vargiu, L.; La Colla, M.; Loddo, R. J. Med. Chem.
2002, 37, 891.
IR (KBr) cmꢁ1: 3368 (NH), 2208 (CN), 1673 (C@O), 1182 (C–O),
832 (Ar–Cl);
LRMS: (ESI Mꢁ1) 359. HRMS: Calcd for C18H14Cl2N2O2; Exact
mass: 360.0432, found (ESI MꢁH) 359.0397.
4.2.46. (E)-2-Cyano-N-(3,4-dichlorobenzyl)-3-(naphthalen-2-
yl)acrylamide (51)
Synthesized using the general procedure as for (27), from N-
(3,4-dichlorobenzyl)propioamide and 2-naphthaldehyde to afford
(52) as a light yellow solid; 40%; 180–181 °C.
1H NMR (DMSO-d6) (300 MHz): d 9.08 (t, J = 5.6 Hz, NH), 8.45 (s,
1H, CH@C), 8.36 (s, 1H, Ar2 H1), 8.14–7.97 (m, 4H, Ar2 H5 + Ar2
H8 + Ar2 H4 + Ar H5), 7.69-7.58 (m, 4H, Ar2 H6 + Ar2 H7 + Ar2
H3 + Ar H1), 7.33 (dd, J = 1.8, 8.3 Hz, 1H, Ar H6), 4.43 (d,
J = 5.6 Hz, 2H, CH2NH);
13C NMR (DMSO-d6) (75 MHz): d 161.2, 151.0, 140.0, 134.3,
132.7, 132.3, 130.8, 130.4, 129.5, 129.4, 129.0, 128.8, 128.6,
127.8, 127.8, 127.7, 127.2, 124.5, 116.4, 105.7, 42.2;
IR (KBr) cmꢁ1: 3370 (NH), 2212 (CN), 1686 (C@O), 1257 (C–O),
739 (Ar–Cl);
RMS: (ESI Mꢁ1) 379. HRMS: Calcd for C21H14Cl2N2O; Exact
15. Carta, A.; Palomba, M.; Boatto, G.; Busonera, B.; Murreddu, M.; Loddo, R. IL
Farmaco 2004, 59, 637–644.
mass: 380.0483, found (ESI MꢁH) 379.0541.
16. Carta, A.; Briguglio, I.; Piras, S.; Boatto, G.; La Colla, P.; Loddo, R.; Tolomeo, M.;
Grimaudo, S.; Cristina, A. D.; Pipitone, R. M.; Laurini, E.; Paneni, M. S.; Posocco,
P.; Fermeglia, M.; Pricl, S. Eur. J. Med. Chem. 2011, 46, 4151.
17. Gazit, A.; Osherov, N.; Gilon, C.; Levitzki, A. J. Med. Chem. 1996, 39, 4905.
18. Ohsumi, K.; Nakagawa, R.; Fukuda, Y.; Hatanaka, T.; Morinaga, Y.; Nihei, Y.;
Osishi, K.; Suga, Y.; Akiyama, Y.; Tsuji, T. J. Med. Chem. 1998, 41, 3022.
19. Quiroga, J.; Cobo, D.; Insuasty, B.; Abonia, R.; Nogueras, M.; Cobo, J.; Vasquez,
Y.; Gupta, M.; Derita, M.; Zacchino, S. Arch. Pharm. 2007, 340, 603.
20. Repicky, A.; Jantova, S.; Cipak, L. Cancer Lett. 2009, 277, 55.
21. Saczewski, F.; Reszka, P.; Gdaneic, M.; Grunert, R.; Bednarski, P. J. J. Med. Chem.
2004, 47, 3449.
22. Saczewski, F.; Stencel, A.; Bienczak, A. M.; Langowska, K.; Michaelis, M.; Werel,
W.; Halasa, R.; Reszka, P.; Bednarski, P. J. Eur. J. Med. Chem. 2008, 43, 1847.
23. Yamazaki, R.; Nishiyama, Y.; Furuta, T.; Hatano, H.; Igarashi, Y.; Asakawa, N.;
Kodaira, H.; Takahashi, H.; Aiyama, R.; Matsuzaki, T.; Yagi, N.; Sugimoto, Y. Mol.
Cancer Ther. 2011, 10, 1252.
4.2.47. (E)-2-Cyano-N-(3,4-dichlorobenzyl)-3-(naphthalen-1-
yl)acrylamide (52)
Synthesized using the general procedure as for (27), from N-
(3,4-dichlorobenzyl)propioamide and 1-naphthaldehyde to afford
(52) as a light yellow solid; 57%; 173–175 °C.
1H NMR (DMSO-d6) (300 MHz): d 9.25 (t, J = 5.7 Hz, NH), 8.90
(s, 1H, CH@C), 8.14–8.02 (m, 4H, Ar2 H5 + Ar2 H4 + Ar2 H8 + Ar2
H2), 7.68–7.59 (m, 5H, Ar2 H3 + Ar2 H6 + Ar2 H7 + Ar H1 + Ar
H5), 7.37 (dd, J = 1.7, 8.3 Hz, 1H, Ar H6), 5.73 (d, J = 5.7 Hz, 2H,
CH2NH);
13C NMR (DMSO-d6) (75 MHz): d 161.0, 148.9, 140.0, 132.9,
131.9, 130.8, 130.7, 130.4, 129.5, 129.4, 129.2, 128.7, 127.8,
127.5, 127.2, 126.8, 125.4, 123.6, 115.9, 110.3, 42.1;
IR (KBr) cmꢁ1: 3370 (NH), 2214 (CN), 1680 (C@O), 779 (Ar–Cl);
LRMS: (ESI Mꢁ1) 379. HRMS: Calcd for C21H14Cl2N2O; Exact
mass: 380.0483, found (ESI MꢁH) 379.0469.
24. Slatt, J.; Bergman, J. Synthesis 2004, 16, 2760.
25. Alberghina, G.; Amato, M. E.; Bottino, F. A.; Corsaro, A.; Fisichella, S. J.
Heterocycl. Chem. 1986, 23, 1747–1752.
26. Herz, W.; Brasch, J. J. Org. Chem. 1958, 23, 711.
27. Wang, K.; Kim, D. D.; Dömling, A. J. Comb. Chem. 2010, 12, 111.
28. Radi, M.; Botta, L.; Casaluce, G.; Bernardini, M.; Botta, M. J. Comb. Chem. 2010,
12, 200.
29. McCluskey, A.; Robinson, P. J.; Hill, T.; Scott, J. L.; Edwards, J. K. Tetrahedron Lett.
2002, 43, 3117.
30. Inokuchi, T.; Kawafuchi, H. J. Org. Chem. 2006, 71, 947.
31. Ryabukhin, R. V.; Plaskon, A. S.; Volochnyuk, D. M.; Pipko, S. E.; Shivanyuk, A.
N.; Tolmachev, A. A. J. Comb. Chem. 2007, 9, 1073.
32. Bergman, A. M.; van Haperen, V. W. R.; Veerman, G.; Kuiper, C. M.; Peters, G. J.
Clin. Cancer Res. 1996, 2, 521.
Acknowledgements
This project was funded by the Australian Research Council. MT
acknowledges the receipt of scholarship support from the Univer-
sity of Newcastle, Australia.
33. Sakoff, J. A.; Ackland, S. P. Cancer Chemother. Pharmacol. 2000, 46, 477.