Parallel solution-phase synthesis of targeted tyrphostin libraries with anticancer activity

Article abstract of DOI:10.1071/CH04143

The combination of semi-automation, an elegant synthesis, and parallel solution-phase synthesis approaches has allowed the development of five targeted, symmetrical tyrphostin compound libraries. These libraries on average are comprised of 12 compounds. Notwithstanding this, low micromolar potent growth inhibitors against HT29 (colorectal carcinoma) and G401 (renal carcinoma) cell lines were discovered. Additionally, significant SAR data was obtained. We noted that the most potent growth inhibitory activity was consistently observed for those analogues that possessed a 2-chlorophenyl (for 10: GI _50HT29 5.5 ± 0.4 μM, GI_50G401 2.6 ± 0.4 μM; for 23: GI_50HT29 2.4 ± 0.2 μM, GI_50G401 1.9 ± 1 μM; for 34: GI_50HT29 8.8 ± 3.1 μM. GI_50G401 6.2 ± 2.9 μM; for 46: GI_50HT29 5.2 ± 0.9 μM, GI_50G401 3.7 ± 0.6 μM; for 57: GI _50HT29 4.6 ± 0.8 μM, GI_50G401 2.1 ± 0.2 μM), a 3-chlorophenyl (for 11: GI_50HT29 3.8 ± 0.7 μM, GI_50G401 1.7 ± 0.7 μM; for 48: GI_50HT29 5.9 ± 0.1 μM, GI_50G401 3.4 ± 0.6 μM; for 58: GI _50HT29 4.8 ± 0.9 μM, GI_50G401 3.4 ± 0.2 μM), or a 3-methoxyphenyl substituent (for 13: GI_50HT29 7.4 ± 3.8 μM, GI_50G401 2.8 ± 0.5 μM; for 26: GI _50HT29 4.5 ± 0.5 μM, GI_50G401 4.9 ± 1 μM; for 37: GI_50HT29 3.7 ± 0.2 μM, GI_50G401 1.6 ± 0.2 μM; for 49: GI_50HT29 3.7 ± 0.4 μM, GI_50G401 3.4 ± 0.2 μM; for 60: GI_50HT29 4.1 ± 0.6 μM, GI_50G401 1.8 ± 0.3 μM). Finally, we noted that increasing the distance between the terminal aromatic rings had only a minimal effect on the 2-, 3-chlorophenyl, and 3-methoxyphenyl analogues, but did have a favourable effect on OH, COOH, and multiply substituted analogues. CSIRO 2005.

Full text of DOI:10.1071/CH04143

Full Paper
CSIRO PUBLISHING
Aust. J. Chem. 2005, 58, 94–103
www.publish.csiro.au/journals/ajc
Parallel Solution-Phase Synthesis of Targeted Tyrphostin Libraries
with Anticancer Activity
A
B
C
A,D
Timothy A. Hill, Jennette A. Sakoff, Phillip J. Robinson, and Adam McCluskey
A
Chemistry, School of Environmental and Life Sciences, University of Newcastle,
Callaghan NSW 2308, Australia.
B
Department of Medical Oncology, Mater Misericordiae Hospital, Waratah NSW 2298, Australia.
Children’s Medical Research Institute, 214 Hawkesbury Road, Westmead NSW 2145, Australia.
Corresponding author. Email: Adam.McCluskey@newcastle.edu.au
C
D
The combination of semi-automation, an elegant synthesis, and parallel solution-phase synthesis approaches has
allowed the development of five targeted, symmetrical tyrphostin compound libraries. These libraries on average
are comprised of 12 compounds. Notwithstanding this, low micromolar potent growth inhibitors against HT29
(colorectal carcinoma) and G401 (renal carcinoma) cell lines were discovered. Additionally, significant SAR data
was obtained. We noted that the most potent growth inhibitory activity was consistently observed for those ana-
logues that possessed a 2-chlorophenyl (for 10: GI50 HT29 5.5 ± 0.4 µM, GI50 G401 2.6 ± 0.4 µM; for 23: GI50 HT29
2
5
.4 ± 0.2 µM, GI50 G401 1.9 ± 1 µM; for 34: GI50 HT29 8.8 ± 3.1 µM, GI50 G401 6.2 ± 2.9 µM; for 46: GI50 HT29
.2 ± 0.9 µM, GI50 G401 3.7 ± 0.6 µM; for 57: GI50 HT29 4.6 ± 0.8 µM, GI50 G401 2.1 ± 0.2 µM), a 3-chlorophenyl
(
for 11: GI50 HT29 3.8 ± 0.7 µM, GI50 G401 1.7 ± 0.7 µM; for 48: GI50 HT29 5.9 ± 0.1 µM, GI50 G401 3.4 ± 0.6 µM;
for 58: GI50 HT29 4.8 ± 0.9 µM, GI50 G401 3.4 ± 0.2 µM), or a 3-methoxyphenyl substituent (for 13: GI50 HT29
7
3
4
.4 ± 3.8 µM, GI50 G401 2.8 ± 0.5 µM; for 26: GI50 HT29 4.5 ± 0.5 µM, GI50 G401 4.9 ± 1 µM; for 37: GI50 HT29
.7 ± 0.2 µM, GI50 G401 1.6 ± 0.2 µM; for 49: GI50 HT29 3.7 ± 0.4 µM, GI50 G401 3.4 ± 0.2 µM; for 60: GI50 HT29
.1 ± 0.6 µM, GI50 G401 1.8 ± 0.3 µM). Finally, we noted that increasing the distance between the terminal aromatic
rings had only a minimal effect on the 2-, 3-chlorophenyl, and 3-methoxyphenyl analogues, but did have a favourable
effect on OH, COOH, and multiply substituted analogues.
Manuscript received: 3 June 2004.
Final version: 25 October 2004.
Introduction
parallel solution-phase synthesis (PSPS). In this particular
instance, we have developed methodologies that allow the
rapid synthesis of bistyrphostin type molecules 1 (Scheme 1).
The tyrphostins are a class of benzylidene malonitrile based
More than five decades of research effort in cancer drug dis-
covery and development have provided only 86 new products
[1]
for the treatment of malignancy. Although major advances
have been made in the chemotherapeutic management of
some patients, particularly in haematologic malignancies,
one-halfofallcancerpatientseitherdonotrespondtotherapy,
or relapse following initial response and ultimately die from
[
3]
compounds developed by Levitski and coworkers. They
have been shown to have a diversity of molecular targets,
for example tyrosine kinases, GTP-using enzymes, calcin-
[
3–6]
urin, and the AP-2 adaptor complex.
From the initial
discovery of tyrphostins, extensive efforts have stemmed
to develop highly potent tyrosine kinase inhibitors such as
STI 571 (Glivec), which became the first signal transduc-
tion inhibitor to show efficacy in the clinic for chronic
[2]
their metastatic disease. For these patients, better systemic
therapy offers the only chance for cure or prolonged survival.
Discovery of such agents requires continued research into
novel therapeutic products that can be used in combination
with biological agents and immune therapies in order to erad-
icate systemic disease not curable by surgery or irradiation.
Understandably, there is considerable pressure to develop
new treatments and therapeutic approaches for the treatment
of cancer. As part of a series of on-going studies within
our laboratories to generate small targeted libraries, which
are the first synthetic iteration in the development of new
therapeutic agents, we have developed a series of simple
synthetic approaches that are amenable to semi-automated,
O
O
N
H
N
H
CN
CN
HO
OH
OH
OH
1
Scheme 1.
10.1071/CH04143
©
CSIRO 2005
0004-9425/05/020094

Synthesis of Targeted Tyrphostin Libraries with Anticancer Activity
95
[
7]
myeloid leukaemia. Bistyrphostin 1 inhibits EGFR tyro-
sine kinase (IC50 0.4 µM) and blocks EGF cell proliferation
the need to develop novel chemical methodologies to obtain
our compound libraries. Secondly, solution-phase synthe-
sis allows compounds to be produced on a larger scale.
Hence, this would allow us to generate around 300 mg of each
sample, and thereby allow us to build a comprehensive library
of compounds for use in several experiments and biological
screens. The main advantage of a solid-phase methodology,
namely the ease of purification, was not an issue with our
tyrphostin libraries because isolation of the final products
was simplified by careful selection of solvents. In particular,
the desired products were insoluble in the solvents chosen
and, therefore, could be isolated by filtration of the reaction
mixtures.
[8]
IC50 3 µM). It has also been reported to inhibit HIV
integrase, as well as voltage-operated calcium channel cur-
rents in vascular smooth muscle cells through inhibition of
(
[9]
[10]
src tyrosine kinase activity.
Given the oncology related
success of Glivec and other tyrosine kinase inhibitors, we
felt it appropriate to explore the potential of bistyrphostin
like compounds developed in our laboratory as anticancer
agents.
Tyrphostins were originally synthesized by traditional
methods that used individual reactions to yield the desired
[3,8]
targets in gramme quantities.
More recently, other groups
have synthesized tyrphostin libraries using solid-phase com-
binatorial synthesis to produce small quantity libraries
Herein we report the parallel, solution-phase synthesis of
compound libraries based upon bistyrphostin 1 and the anti-
tumour activity of these libraries in two human cancer cell
lines.
[11,12]
(
100 mg) of up to 4500 compounds.
However, we feel
that while there have been significant advances in solid-phase
synthesis, it is still not routinely available for small labo-
ratories that require specialized techniques, equipment, and
approaches. The added cost of materials used in the synthe-
ses, and inevitable wastage, is also a drawback. For these
reasons, solution-phase chemistry was deemed to be a more
desirable option to be pursued as it has a more traditional
organic chemistry basis.
Results and Discussion
In this preliminary study towards the development of targeted
libraries of tyrphostins, we pursued multiple modifications of
1
5
both the aromatic nucleus (R –R ) and the linker (X) in 1.
This is shown schematically in Scheme 2.
We and others have previously dissected molecules of
this type and developed synthetic pathways to a variety of
The advantages of using solution-phase chemistry in the
development of our bistyrphostin based libraries were two-
fold. Firstly, application of the Knoevenagel condensation
reaction in a similar manner to that used by Levitski in the
[
8,13]
targets.
In this work, we commenced from commer-
cially available 1,2-diaminoethane 3a, which was allowed
to react in a solvent-free medium with methyl cyanoacetate
[8]
original synthesis of dimeric tyrphostins would alleviate
2
to afford the corresponding biscyanoamide 4a in excel-
lent yield (>90%, Scheme 3). Next, in order to expedite
our library development, we conducted a Knoevenagel con-
densation with a small selection of commercially available
benzaldehydes (see Table 1 for details). To further accelerate
the library development, these manipulations were conducted
using the Büchi Syncore reaction station; this allowed semi-
automation of the process. Thus, 12 of the 24 reaction wells
available were charged with one equivalent of 3a in ethanol
and a catalytic amount of piperidine, and to these were added
R5
R5
O
O
R4
R4
X
N
H
N
H
CN
CN
R
R3
R1
1
R3
R2
R2
Scheme 2.
O
O
H
N
(a)
NH2
NC
ϩ
NC
H2N
(
)n
N
H
( )n
CN
OMe
O
2
3
4a–4d
a n ϭ 0; b n ϭ 1;
c n ϭ 2; d n ϭ 3
R5
R4
R3
(b)
O
R1
R2
R2
R1
R3
R5
O
CN
H
N
R4
R3
R4
N
H
( )n
R5
CN
O
R1
R2
5–63
Scheme 3. Reagents and conditions: (a) mix, solvent free, 5 min at room temperature; (b) piperidine (cat.), (substituted)benzaldehyde, ethanol,
reflux, 2 h.

9
6
T. A. Hill et al.
twoequivalentsofanappropriatebenzaldehyde.Themixtures
were then heated at reflux for two hours, allowed to cool, and
were subsequently filtered, so that the capabilities of the Syn-
core were exploited. The resultant solids were washed with
diethyl ether, refiltered, and dried in situ. Subsequent analysis
showed that all the products were greater than 95% pure by
NMR spectroscopy.
Upon having successfully accomplished the rapid syn-
thesis of our first 12-compound library, we screened the
compounds against two cancer cell lines at 50 µM drug
concentrations; HT29 (colorectal carcinoma) and G401
GIHT29 58 ± 16%, GIG401 72 ± 21%; respectively at 50 µM
drug concentration). These data suggest that either a
2-, 3-chlorophenyl, or a 3-methoxyphenyl substituent plays a
positive role in inhibiting cell growth in this type of analogue.
Table 1. Growth inhibition of library 1 against HT29 (colorectal
carcinoma) and G401 (renal carcinoma) cell lines
GI [%] is a measure of the percentage growth inhibition relative to
untreated control cells after a 72 h exposure to 50 µM of the test
compound. As presented, higher values indicate greater growth
inhibition. A value greater than 100% is indicative of complete growth
inhibition together with cell death.
(
renal carcinoma). As shown in Table 1, all compounds
except 12 and 15 displayed growth inhibition of greater
than 50% at this concentration, while compounds, 10, 11,
and 13 are of particular interest as they showed total
growth inhibition of greater than 100%. More detailed
analysis afforded GI50 values in the low micromolar
range (10: GI50 HT29 5.5 ± 0.4 µM, GI50 G401 2.6 ± 0.4 µM;
R2
R1
CN
R3
R5
O
H
N
R4
R4
N
H
R5
CN
O
R3
R1
1
1: GI50 HT29 3.8 ± 0.7 µM, GI50 G401 1.7 ± 0.1 µM;
3: GI50 HT29 7.4 ± 3.8 µM, GI50 G401 2.8 ± 0.5 µM).
Both 10 and 11 possess a single chloride substituent
R2
1
Compound R1 R2
R3
R4 R5 HT29 GI G401 GI
in the 2- and 3-position of the aromatic ring, respectively.
It is notable that the corresponding 4-chlorophenyl ana-
logue 12 is significantly less potent at inhibiting cell growth
[%]
[%]
5
6
7
8
9
1
1
12
13
1
1
1
H
H
H
H
H
Cl
H
H
H
H
H
H
H
OH
H
OH
OMe OMe
H
Cl
H
OMe
H
H
H
OH
OH
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
73 ± 4
61 ± 12
53 ± 10
57 ± 19
56 ± 8
96 ± 15
75 ± 4
(
GIHT29 34 ± 5% and GIG401 67 ± 5%). This apparent dis-
66 ± 4
72 ± 12
68 ± 4
favour of the 4-position is more apparent when the data
obtained for the chloride-substituted species is examined in
conjunction with that of the other active compound, namely
0
1
H
H
Cl
H
OMe
OMe
COOH
107 ± 4
115 ± 7
34 ± 5
105 ± 1
110 ± 5
67 ± 5
3-methoxyphenyl 13. The corresponding 4-methoxy 14,
3,4-dimethoxy 9, 3-hydroxy-4-methoxy 15, and 4-carboxy
16 compounds all display significantly poorer growth inhi-
104 ± 3
62 ± 3
106 ± 2
62 ± 2
4
5
6
OH
H
47 ± 8
74 ± 19
72 ± 21
bition (GIHT29 62 ± 3%, GIG401 62 ± 2%; GIHT29 56 ± 8%,
58 ± 16
GIG401 68 ± 4%; GIHT29 47 ± 8%, GIG401 74 ± 19%; and
Table 2. Growth inhibition of library 2 against HT29 (colorectal carcinoma) and G401 (renal carcinoma) cell lines
GI [%] is a measure of the percentage growth inhibition relative to untreated control cells after a 72 h exposure to 50 µM of
the test compound. As presented, higher values indicate greater growth inhibition. A value greater than 100% is indicative of
complete growth inhibition together with cell death. GI50 is the concentration of the test drug required to inhibit the cell
growth by 50%.
R5
R5
O
O
R4
R3
R4
R3
N
H
N
H
CN
CN
R1
R1
R2
R2
Compound R1 R2
R3
R4
R5 HT29 GI [%] G401 GI [%] HT29 GI50 [µM] G401GI50 [µM]
17
18
19
20
21
22
23
24
25
26
27
28
29
H
H
H
H
H
H
Cl
H
H
H
H
H
H
H
OH
H
OH
OH
OMe OH
H
Cl
H
OMe
H
H
H
OH
OH
OH
H
H
H
H
OH
OH
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
110 ± 5
110 ± 3
89 ± 4
55 ± 5
48 ± 8
27 ± 9
111 ± 3
102 ± 5
110 ± 3
113 ± 5
54 ± 3
94 ± 6
37 ± 5
106 ± 2
105 ± 1
94 ± 3
67 ± 6
88 ± 7
52 ± 12
107 ± 1
94 ± 4
107 ± 1
105 ± 1
66 ± 4
85 ± 4
30 ± 6
12 ± 7.4
9.7 ± 0.3
21 ± 3.1
43 ± 3.3
58 ± 4.4
103 ± 23
2.4 ± 0.2
28 ± 8.3
5.0 ± 0.6
4.5 ± 0.5
41 ± 9.8
32 ± 2.8
82 ± 19
6.7 ± 1.5
5.7 ± 1.4
11 ± 1.9
30 ± 8.2
25 ± 4.4
45 ± 10
1.9 ± 1
H
H
Cl
H
OMe
OMe
COOH
14 ± 0.7
5.1 ± 1.2
4.9 + 1
24 ± 8
OH
H
23 ± 3.3
80 ± 12

Synthesis of Targeted Tyrphostin Libraries with Anticancer Activity
97
Table 3. Growth inhibition of libraries 3–5 against HT29 (colorectal carcinoma) and G401 (renal carcinoma) cell lines
GI [%] is a measure of percentage growth inhibition relative to untreated control cells after a 72 h exposure to 50 µM of the test
compound. As presented, higher values indicate greater growth inhibition. A value greater than 100% is indicative of complete
growth inhibition together with cell death. GI50 is the concentration of test drug required to inhibit the cell growth by 50%.
R5
O
O
R5
R4
R3
R4
R3
N
H
N
H
n
CN
CN
R1
R1
R2
R2
Compound R1 R2
Library 3
R3
R4 R5
n
HT29 GI [%] G401 GI [%] HT29 GI50 [µM] G401 GI50 [µM]
3
3
3
3
3
3
3
3
3
3
4
0
1
2
3
4
5
6
7
8
9
0
H
H
H
H
Cl
H
H
H
H
H
H
H
OH
H
OH
H
Cl
H
OMe
H
OH
H
H
H
OH
OH
H
H
Cl
H
OMe
OMe
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
1
1
1
1
1
1
1
1
1
1
1
100 ± 7
98 ± 7
42 ± 7
46 ± 7
114 ± 7
69 ± 7
66 ± 7
103 ± 7
35 ± 7
39 ± 7
92 ± 7
107 ± 7
104 ± 7
57 ± 7
82 ± 7
110 ± 7
103 ± 7
89 ± 7
105 ± 7
62 ± 7
55 ± 7
99 ± 7
2.9 ± 0.2
2.6 ± 0.4
6.5 ± 0.3
3.4 ± 0.8
–
–
–
–
8.8 ± 3.1
6.2 ± 2.9
28 ± 8.3
14 ± 0.7
–
–
3.7 ± 0.2
1.6 ± 0.2
–
–
–
–
–
–
COOH
Library 4
4
4
4
4
4
4
4
4
4
5
5
5
1
2
3
4
5
6
7
8
9
0
1
2
H
H
H
H
H
Cl
H
H
H
H
H
H
H
OH
H
OH
OMe OMe
H
Cl
H
OMe
H
H
H
OH
OH
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
2
2
2
2
2
2
2
2
2
2
2
2
111 ± 7
99 ± 7
106 ± 7
107 ± 7
82 ± 7
4.9 ± 0.3
13 ± 5.9
–
3.2 ± 0.2
3.9 ± 1
–
49 ± 7
102 ± 7
42 ± 7
101 ± 7
81 ± 7
21 ± 2.9
6.2 ± 0.4
–
–
H
H
Cl
H
OMe
OMe
COOH
112 ± 7
117 ± 7
110 ± 7
110 ± 7
86 ± 7
106 ± 7
106 ± 7
108 ± 7
109 ± 7
89 ± 7
5.2 ± 0.9
7 ± 1.5
5.9 ± 0.1
3.7 ± 0.4
–
3.7 ± 0.6
4.7 ± 0.4
3.4 ± 0.6
3.2 ± 0.2
–
OH
H
102 ± 7
104 ± 7
94 ± 7
8.3 ± 1.2
3.4 ± 0.3
109 ± 7
23 ± 6.1
8.2 ± 1.2
Library 5
53
54
55
56
57
58
59
60
61
62
63
H
H
H
H
Cl
H
H
H
H
H
H
H
OH
H
OH
H
Cl
H
OMe
H
OH
H
H
H
OH
OH
H
H
Cl
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
3
3
3
3
3
3
3
3
3
3
3
107 ± 7
72 ± 7
64 ± 7
54 ± 7
107 ± 7
76 ± 7
66 ± 7
105 ± 7
28 ± 7
59 ± 7
36 ± 7
107 ± 7
84 ± 7
79 ± 7
70 ± 7
106 ± 7
99 ± 7
90 ± 7
105 ± 7
46 ± 7
77 ± 7
53 ± 7
6.4 ± 1.8
6.0 ± 0.6
–
–
–
–
–
–
4.6 ± 0.8
2.1 ± 0.2
4.8 ± 0.9
3.4 ± 0.2
–
–
4.1 ± 0.6
1.8 ± 0.3
OMe
OMe
COOH
–
–
–
–
–
–
= GI50 not determined.
Also of note in this initial library is that neither of the free
hydroxy analogues, 6 nor 7, are comparatively potent cell
growth inhibitors (GIHT29 61 ± 12%, GIG401 75 ± 4%; and
GIHT29 53 ± 10%, GIG401 66 ± 4%; respectively).
of small-targeted chain elongation libraries of between 10
and 12 compounds.
Insertion of an additional methylene group proved to be
a trivial exercise. Simple substitution of 1,2-diaminoethane
(from the synthesis of library 1) with 1,3-diaminopropane
afforded the corresponding propane-bridged biscyanoamido-
acetate 3b in excellent yield (>90%). Subsequent manipula-
tions were conducted as described previously, and once again,
excellent yields and purities were obtained. Library 2 ana-
logues were then screened against both HT29 and G401 cell
lines at a single 50 µM compound dose, as well as at multiple
doses in order to produce a dose–response curve from which
Given the apparent roles of both Cl and OMe in library
1
, we were keen to explore other simple modifications that
might be expected to have an impact on growth inhibi-
tion. In considering potential modifications, we took into
account the ease of compound manipulation using our syn-
thetic approach (Scheme 3) and the utility (and applicability)
of the Syncore system. Thus, in this preliminary report we
will limit the chemistry and subsequent discussion to a series

9
8
T. A. Hill et al.
a GI50 value could be calculated. These data are presented in
Table 2.
is ongoing in our laboratories and new developments will be
reported in due course.
A preliminary analysis of the percentage growth inhibi-
tion values at 50 µM highlighted compounds 17 and 18, as
well as 23–26 as being of particular interest (>90% growth
inhibition). Moreover, at least one cell line registered com-
plete growth inhibition (>100%). Closer examination of the
substituent patterns revealed differences between libraries
Experimental
Biological Evaluation
Cell Culture and Stock Solutions
A 10 mM stock solution in 100% dimethyl sulfoxide [(CD3)2SO]
was prepared for each test compound and stored at 4 C. Further dilu-
◦
1
1
1
2
and 2. Most notable are the free phenyl 17 (GI50 HT29
2 ± 7.4 and GI50 G401 6.7 ± 1.5 µM), the 3-hydroxyphenyl
8 (GI50 HT29 9.7 ± 0.3 and GI50 G401 5.7 ± 1.4 µM), and the
- and 4-chlorophenyl 23 and 25 (GI50 HT29 2.4 ± 0.2 and
tions were made using cell culture media (DMEM, Trace Biosciences)
and the final DMSO content was <0.5%. The HT29 (human colon car-
cinoma) cell line was maintained in DMEM supplemented with 10 mM
sodium bicarbonate, while the G401 (human kidney carcinoma) cell
line was maintained in McCoys (Trace Biosciences). All culture media
was further supplemented with fetal bovine serum (10%), penicillin
GI50 G401 1.9 ± 1 µM; and GI50 HT29 5.0 ± 0.6 and GI50 G401
−
1
−1
5
.1 ± 1.2 µM, respectively), the last chlorophenyl analogues
(100 IU mL ), streptomycin (100 µg mL ), and glutamine (4 mM).
Cells were passaged every 3–7 days and all cell lines were routinely
tested and found to be mycoplasma-free. Both cell lines were cultured
being more potent than the corresponding 3-chlorophenyl
analogue 24 (GI50 HT29 28 ± 8.3 and GI50 G401 14 ± 0.7 µM).
However, the previous observation of excellent growth inhi-
bition associated with the 3-methoxyphenyl analogue 13 was
still observed with the methylene extended 26 (GI50 HT29
◦
in air at 37 C under 5% CO2.
Cytotoxicity Assay
Cells in logarithmic growth were transferred to 96-well plates. Cyto-
toxicity was determined by plating cells in triplicate in 100 µL of
medium at a density of 2500–3500 cells per well for each cell line.
On day zero (24 h after plating), when the cells were in logarith-
mic growth, 100 µL of medium with or without the test agent was
added to each well. After drug exposure for 72 h, growth inhibitory
effects were evaluated using the MTT [3-(4,5-dimethyltiazol-2-yl) 2,5-
diphenyltetrazolium bromide] assay and absorbance was read at 540 nm.
All drugs were initially evaluated at a single drug concentration of
4
.5 ± 0.5andGI50 G401 4.9 ± 1 µM), andtoalesserextent, the
corresponding 4-methoxy analogue 27 (GI50 HT29 41 ± 9.8
and GI50 G401 24 ± 8 µM). It is also noteworthy that two
oxygen-bearing substituents (20–22 and 28) could be tol-
erated and were essentially equipotent with 27.
Our final series of synthetic manipulations involved addi-
tional methylene insertions, and each subsequent library was
synthesized from 1,4-diaminobutane, 1,5-diaminopentane,
and 1,6-diaminohexane, respectively. All synthetic proce-
dures were conducted using the Syncore station as before,
and all compounds were synthesized rapidly in high yield
and purity. The results of screening against HT29 and G401
carcinoma cell lines are given in Table 3.
5
0 µM, from which a GI (%) value was calculated that represented the
percentage cell growth inhibition. Compounds that induced appreciable
growth inhibition underwent a multiple dose–response assessment from
which an IC50 value, which represents the drug concentration (µM) that
induces 50% growth inhibition, was calculated. All calculations were
based on the difference between the optical density values on day zero
and those at the end of the drug exposure period.
Examination of the data presented in Table 3 does
not show any major improvements in growth inhibi-
tion. Modest improvements in growth inhibition were
noted for the free phenyl analogues upon insertion of an
additional methylene (compare 17 GI50 HT29 12 ± 7.4 µM,
GI50 G401 6.7 ± 1.5 µM with 30 GI50 HT29 2.9 ± 0.2 µM,
GI50 G401 2.6 ± 0.4 µM). However, subsequent methylene
insertions had almost no further effect on growth inhibi-
tion (41: GI50 HT29 4.9 ± 0.3 µM, GI50 G401 3.2 ± 0.2 µM;
General
All starting materials were purchased from Aldrich Chemical Co.
and Lancaster Synthesis. 1H and 13C NMR spectra were recorded
on a Bruker Advance AMX 300 MHz spectrometer at 300.1315 and
75.4762 MHz, respectively. Chemical shifts are relative to trimethylsi-
lane (TMS) as internal standard. All compounds returned satisfactory
analyses.
Synthetic Methods
2
-Cyano-N-[3-(2-cyanoacetylamino)ethyl]acetamide 4a
5
3: GI50 HT29 6.4 ± 1.8 µM, GI50 G401 6.0 ± 0.6 µM). It
Ethylenediamine 3a (1.5 g, 25 mmol) and methyl cyanoacetate 2
should be noted that compound 44 was unusual as it dis-
played considerably improved potency despite the fact that
methylene insertions to give 8, 20, and 33 had little effect.
(
5 g, 50 mmol) were stirred at room temperature for 2 h. The resultant
white solid was then mixed with ethanol (10 mL) and collected by fil-
tration. Recrystallization from ethanol gave a white solid (6.3 g, 81%),
◦
[8]
◦
mp 182 C (lit. 183 C). δH [(CD3)2SO] 8.25 (2H, t, J 5.5), 3.56 (4H,
s), 3.13 (4H, br s). δC [(CD3)2SO] 162.3, 115.9, 38.4, 25.3.
Conclusions
2
-Cyano-N-[3-(2-cyanoacetylamino)propyl]acetamide 4b
We have developed a facile approach to the rapid synthe-
sis of small-targeted libraries of bioactive compounds. The
combination of a good lead compound, simple synthesis, and
parallel solution-phase synthesis has allowed the rapid iden-
tification of greater than 20 analogues with potent growth
inhibition in two cancer derived cell lines, HT29 and G401.
This work highlights the utility of PSPS approaches as well
as validating the use of small libraries (rather than libraries
of hundreds if not thousands of compounds) in the develop-
ment of more potent bioactive lead compounds. This work
Propanediamine 3b (2.2 g, 30 mmol) and methyl cyanoacetate 2
6.4 g, 65 mmol) were stirred at room temperature for 2 h. The resul-
(
tant white solid was then mixed with ethanol (20 mL) and collected by
filtration. Recrystallization from ethanol gave a white solid (5.0 g, 81%),
◦
[8]
◦
mp 146 C (lit. 148 C). δH [(CD3)2SO] 8.21 (2H, t, J 5.5), 3.59 (4H,
s), 3.07 (4H, q, J 6.7), 1.53 (2H, quin, J 6.7). δC [(CD3)2SO] 162.5,
1
16.6, 39.3, 28.9, 25.7.
2-Cyano-N-[3-(2-cyanoacetylamino)butyl]acetamide 4c
1
,4-Diaminobutane 3c (3.0 g, 34 mmol) and methyl cyanoacetate 2
(7.0 g, 70 mmol) were stirred at room temperature for 2 h, after which

Synthesis of Targeted Tyrphostin Libraries with Anticancer Activity
time a white solid was formed. The solid was then mixed with ethanol
99
br s). δC [(CD3)2SO] 162.5, 151.0, 161.6, 146.2, 125.8, 123.5, 117.7,
(
10 mL) and collected by filtration. Recrystallization from ethanol gave
116.5, 116.3, 100.8, 39.6.
◦
[8]
◦
a white solid (6.0 g, 78%), mp 145 C (lit. 145 C). δH [(CD3)2SO]
.15 (2H, t, J 5.5), 3.56 (4H, s), 3.05 (4H, br s), 1.38 (4H, br s).
δC [(CD3)2SO] 161.8, 116.1, 38.6, 26.1, 25.2.
8
2-Cyano-N-{2-[2-cyano-3-(3,4-dimethoxyphenyl)-
acryloylamino]ethyl}-3-(3,4-dimethoxyphenyl)acrylamide 9
This was synthesized in the manner described for 5 from 4a and
2
-Cyano-N-[3-(2-cyanoacetylamino)pentyl]acetamide 4d
3
,4-dimethoxybenzaldehyde to yield a light yellow solid (69%), mp >
◦
1
,5-Diaminopentane 3d (2.0 g, 20 mmol) and methyl cyanoacetate
300 C. δH [(CD3)2SO] 8.43 (2H, t, J 5.5), 8.08 (2H, s), 7.65 (2H, d,
J 1.9), 7.55 (2H, dd, J 9.0 and 2.0), 7.14 (2H, d, J 8.6), 3.84 (6H, s),
3.77 (6H, s), 3.31 (4H, br s). δC [(CD3)2SO] 162.1, 152.9, 150.9, 149.1,
125.8, 124.9, 117.5, 112.7, 112.3, 103.0, 56.2, 55.9, 40.2.
2
(3.9 g, 40 mmol) were stirred at room temperature for 2 h, after which
time a white solid was formed. The solid was then mixed with ethanol
10 mL) and collected by filtration. Recrystallization from ethanol gave
a white solid (4.62 g, 98%), mp 125 C (lit. 125 C). δH [(CD3)2SO]
.14 (2H, t, J 5.4), 3.55 (4H, s), 3.03 (4H, q, J 6.4), 1.39 (4H, quin,
J 7.0), 1.23 (2H, quin, J 7.0). δC [(CD3)2SO] 161.8, 116.1, 38.8, 28.3,
5.2, 23.4
(
◦
[8]
◦
3
2
-(2-Chlorophenyl)-N-{2-[3-(2-chlorophenyl)-
-cyanoacryloylamino]ethyl}-2-cyanoacrylamide 10
8
2
This was synthesized in the manner described for 5 from 4a and
◦
2
[
-chlorobenzaldehyde to yield a yellow solid (83%), mp 248–250 C. δH
2
-Cyano-N-[3-(2-cyanoacetylamino)hexyl]acetamide 4e
,6-Diaminohexane 3e (3.0 g, 26 mmol) and methyl cyanoacetate 2
(CD3)2SO] 8.70 (2H, br t, J 5.6), 8.36 (2H, s), 8.00 (2H, dd, J 6.2 and
2.1), 7.62, (2H, m), 7.54 (4H, m), 3.41 (4H, br s). δC [(CD3)2SO] 160.4,
1
(
6.0 g, 60 mmol) were stirred at room temperature for 2 h, after which
time a white solid was formed. The solid was then mixed with ethanol
10 mL) and collected by filtration. Recrystallization from ethanol gave
146.8, 134.0, 133.2, 130.3, 130.0, 129.5, 127.7, 115.3, 110.4, 39.1.
3
-(3-Chlorophenyl)-N-{2-[3-(3-chlorophenyl)-
(
◦
[8]
◦
2-cyanoacryloylamino]ethyl}-2-cyanoacrylamide 11
awhitesolid(6.2 g, 95%), mp141 C(lit. 140 C). δH [(CD3)2SO]8.15
2H, t, J 5.5), 3.56 (4H, s), 3.04 (4H, q, J 6.1), 1.37 (4H, quin, J 5.9),
(
1
This was synthesized in the manner described for 5 from 4a and
.24 (4H, br s). δC [(CD3)2SO] 161.8, 116.1, 38.9, 28.6, 25.8, 25.2
3
1
1
-chlorobenzaldehyde to yield a light yellow solid (70%), mp 172–
74 C. δH [(CD3)2SO] 8.39 (2H, t, J 5.3), 7.97 (2H, s), 7.85 (2H, t, J
.9), 7.86 (2H, m), 7.60 (4H, m), 3.45 (4H, br s). δC [(CD3)2SO] 160.9,
◦
General Procedure for the Synthesis of Targeted Libraries
Each library was prepared from 0.3 M ethanolic stock solutions of
148.1, 133.8, 133.7, 131.6, 131.0, 129.0, 115.6, 108.5, 39.5.
each cyanoacetamide (4a–4e) that contained piperidine (catalytic, 3
drops): 4a library 1; 4b library 2; 4c library 3; 4d library 4; and 4e
library 5. Each Syncore reaction tube was charged with an aliquot of
the appropriate stock solution (5 mL) followed by the appropriate ben-
zaldehyde dissolved in ethanol (5 mL). An example of this is given in
the synthesis of 5, below.
3
2
-(4-Chlorophenyl)-N-{2-[3-(4-chlorophenyl)-
-cyanoacryloylamino]ethyl}-2-cyanoacrylamide 12
This was synthesized in the manner described for 5 from 4a and
-chlorobenzaldehyde to yield a yellow solid (76%), mp 244–246 C.
δH [(CD3)2SO] 8.58 (2H, br t, J 5.5), 8.16 (2H, s), 7.91 (4H, d, J 8.6),
◦
4
7
1
.63 (4H, d, J 6.8), 3.32 (4H, br s). δC [(CD3)2SO] 161.0, 149.1, 136.8,
31.6, 130.7, 129.3, 116.0, 106.9, 39.2.
2
3
-Cyano-N-[2-(2-cyano-3-phenylacryloylamino)ethyl]-
-phenylacrylamide 5
2
-Cyano-N-{2-[2-cyano-3-(3-methoxyphenyl)-
An aliquot of the stock solution generated from 2-cyano-N-[3-(2-
cyanoacetylamino)ethyl]acetamide 4a (5 mL) was placed in a Syncore
reaction tube followed by benzaldehyde (0.33 g, 3.0 mmol) and ethanol
acryloylamino]ethyl}-3-(3-methoxyphenyl)acrylamide 13
This was synthesized in the manner described for 5 from 4a and
3-methoxybenzaldehyde to yield a yellow solid (65%), mp 118–120 C.
[(CD ) SO] 8.57 (2H, br t, J 5.4), 8.14 (2H, s), 7.49 (2H, m), 7.47
(2H, m), 7.15 (4H, m), 3.79 (6H, br s), 3.98 (4H, br s). δC [(CD ) SO]
161.2, 159.4, 150.4, 133.1, 130.3, 122.3, 118.1, 116.3, 114.7, 106.5,
◦
(
5 mL). This mixture was then heated at reflux for 2 h, cooled, filtered,
and washed with cold diethyl ether (10 mL) to afford a yellow solid
δ
H
3 2
◦
(
0.45 g, 79%), mp 178–180 C. δH [(CD3)2SO] 8.57 (2H, br t, J 5.5),
3 2
8
.17 (2H, s), 7.92 (4H, m), 7.57 (6H, m), 3.42 (4H, br s). δC [(CD3)2SO]
1
61.2, 150.5, 132.2, 131.3, 129.9, 129.1, 116.3, 106.4, 38.9.
55.2, 39.4.
2
-Cyano-N-{2-[2-cyano-3-(3-hydroxyphenyl)-
2-Cyano-N-{2-[2-cyano-3-(4-methoxyphenyl)acryloylamino]-
ethyl}-3-(4-methoxyphenyl)acrylamide 14
acryloylamino]ethyl}-3-(3-hydroxyphenyl) acrylamide 6
Synthesized in the manner described for 5 from 4a and 3-
hydroxybenzaldehyde to yield a yellow solid (85%), mp 290–292 C.
δH [(CD3)2SO] 8.54 (2H, br t, J 5.3), 8.04 (2H, s), 7.36 (4H, m), 7.32
This was synthesized in the manner described for 5 from 4a and
◦
◦
4-methoxybenzaldehyde to yield a yellow solid (94%), mp 246–248 C.
H
δ [(CD ) SO] 8.43 (2H, br t, J 5.5), 8.09 (2H, s), 7.95 (4H, d, J 8.9),
3 2
(
1
2H, m), 6.97 (2H, m), 3.38 (4H, br s). δC [(CD3)2SO] 161.8, 158.2,
51.0, 133.5, 130.7, 121.8, 120.1, 116.7, 116.2, 106.5, 39.6.
7.12 (4H, d, J 8.9), 3.84 (6H, s), 3.36 (4H, br s). δC [(CD ) SO] 163.0,
162.1, 150.5, 132.8, 124.8, 117.4, 115.2, 103.1, 56.0, 40.2.
3 2
2
-Cyano-N-{2-[2-cyano-3-(4-hydroxyphenyl)acryloylamino]-
2-Cyano-N-{2-[2-cyano-3-(3-hydroxy-4-methoxyphenyl)-
acryloylamino]ethyl}-3-(3-hydroxy-4-methoxyphenyl)-
acrylamide 15
ethyl}-3-(4-hydroxyphenyl)acrylamide 7
This was synthesized in the manner described for 5 from 4a and
-hydroxybenzaldehyde to yield a yellow solid (78%), mp > 300 C. δH
◦
4
This was synthesized in the manner described for 5 from 4a and
[
(
(CD3)2SO] 8.36 (2H, br t, J 5.5), 8.02 (2H, s), 7.85 (4H, d, J 7.2), 6.89
4H, d, J 8.8), 3.36 (4H, br s). δC [(CD3)2SO] 162.3, 150.8, 133.2, 123.2,
3-hydroxy-4-methoxybenzaldehydetoyieldabrightyellowsolid(98%),
◦
mp 274–276 C. δH [(CD ) SO] 8.40 (2H, br t, J 5.6), 7.97 (2H, s), 7.53
3 2
1
17.6, 116.7, 101.5, 39.6.
(2H, d, J 2.0), 7.37 (4H, dd, J 8.6 and 2.1), 7.06 (4H, d, J 8.6), 3.84 (6H,
s), 3.36 (4H, br s). δH [(CD3)2SO] 161.8, 151.7, 150.3, 146.7, 124.5,
116.8, 115.4, 112.0, 102.2, 55.6, 39.1.
2
-Cyano-N-{3-[2-cyano-3-(3,4-dihydroxyphenyl)-
acryloylamino]ethyl}-3-(3,4-dihydroxyphenyl) acrylamide 8
2
-Cyano-N-{2-[2-cyano-3-(4-carboxyphenyl)acryloylamino]-
This was synthesized in the manner described for 5 from 4a and
◦
ethyl}-3-(4-carboxyphenyl)acrylamide 16
3
(
(
,4-dihydroxybenzaldehyde to yield a yellow solid (81%), mp 290 C
[8]
◦
lit. 295 C). δH [(CD3)2SO] 8.32 (2H, t, J 5.5), 7.92 (2H, s), 7.53
2H, d, J 2.1), 7.25 (2H, dd, J 8.2 and 2.1), 6.85 (2H, d, J 2.1), 3.45 (4H,
This was synthesized in the manner described for 5 from 4a and
4-carboxybenzaldehyde to yield a yellow solid (96%), mp > 300 C.
◦

1
00
T. A. Hill et al.
δH [(CD3)2SO] 8.68 (2H, t, J 5.3), 8.21 (2H, s), 8.00 (4H, d, J 8.0),
3-(4-Chlorophenyl)-N-{3-[3-(4-chlorophenyl)-
7
.97 (4H, d, J 8.1), 3.40 (4H, br s). δC [(CD3)2SO] 166.7, 162.4, 161.0,
2-cyanoacryloylamino]propyl}-2-cyanoacrylamide 24
1
49.4, 135.3, 134.3, 129.8, 129.8, 116.1, 115.9, 108.2.
This was synthesized in the manner described for 5 from 4b and
-chlorobenzaldehyde to yield a yellow solid (81%), mp 156–158 C.
◦
3
2
-Cyano-N-[3-(2-cyano-3-phenylacryloylamino)-
δ
H
[(CD ) SO] 8.49 (2H, t, J 5.6), 8.15 (2H, s), 7.95 (2H, t, J 1.8),
3 2
propyl]-3-phenylacrylamide 17
7.85 (2H, m), 6.96 (4H, m), 3.27 (4H, q, J 6.7), 1.76 (2H, quin, J 6.8).
δC [(CD3)2SO] 160.4, 148.9, 133.9, 133.7, 131.7, 131.0, 130.9, 129.2,
128.3, 115.8, 107.9, 37.6, 28.4.
This was synthesized in the manner described for 5 from 4b and ben-
zaldehyde to yield a yellow solid (81%), mp 138–140 C. δH [(CD3)2SO]
◦
8
(
1
.37 (2H, s), 7.95 (4H, d, J 6.8), 7.52 (6H, m) 7.12 (2H, br t, J 5.5), 3.53
4H, q, J 6.0), 1.70 (2H, br quin, J 6.1). δC [(CD3)2SO] 161.1, 153.2,
32.7, 131.7, 130.6, 129.2, 116.8, 103.8, 37.1, 29.5.
3
-(4-Chlorophenyl)-N-{3-[3-(4-chlorophenyl)-2-
cyanoacryloylamino]propyl}-2-cyanoacrylamide 25
This was synthesized in the manner described for 5 from 4b and
◦
4
-chlorobenzaldehyde to yield a yellow solid (87%), mp 168–172 C.
2
-Cyano-N-{3-[2-cyano-3-(3-hydroxyphenyl)acryloylamino]-
δH [(CD3)2SO] 8.47 (2H, t, J 5.3), 8.15 (2H, s), 7.92 (4H, d, J 8.4), 7.60
4H, d, J 8.4), 3.29 (4H, q, J 5.9), 1.76 (2H, quin, J 6.5). δC [(CD3)2SO]
propyl}-3-(3-hydroxyphenyl)acrylamide 18
(
1
This was synthesized in the manner described for 5 from 4b and
-hydroxybenzaldehyde to yield a yellow solid (71%), mp 212–216 C.
60.6, 149.1, 136.8, 131.6, 130.7, 129.2, 116.1, 106.8, 37.5, 28.5.
◦
3
δH [(CD3)2SO] 8.43 (2H, t, J 5.5), 8.05 (2H, s), 7.35 (4H, m), 7.33
2H, m), 6.96 (2H, m), 3.26 (4H, q, J 6.1), 1.75 (4H, quin, J 6.8).
δC [(CD3)2SO] 161.4, 158.2, 151.0, 133.5, 130.7, 121.8, 120.1, 116.7,
2-Cyano-N-{3-[2-cyano-3-(3-methoxyphenyl)acryloylamino]-
propyl}-3-(3-methoxyphenyl)acrylamide 26
(
This was synthesized in the manner described for 5 from 4b and
-methoxybenzaldehyde to yield a yellow solid (59%), mp 132–134 C.
1
16.2, 106.3, 37.9, 29.1.
◦
3
δH [(CD3)2SO] 8.46 (2H, t, J 5.4), 8.14, (1H, s), 7.50 (2H, s), 7.44 (4H,
m), 7.13 (2H, t, J 7.5), 3.78 (6H, s), 3.29 (4H, q, J 5.6), 1.77 (2H, quin,
2
-Cyano-N-{3-[2-cyano-3-(4-hydroxyphenyl)acryloylamino]-
propyl}-3-(4-hydroxyphenyl)acrylamide 19
J 6.3). δC [(CD ) SO] 160.8, 159.3, 150.4, 133.1, 130.2, 122.3, 118.1,
3 2
1
16.3, 106.4, 55.2, 37.5, 28.5.
This was synthesized in the manner described for 5 from 4b and
-hydroxybenzaldehyde to yield a yellow solid (58%), mp 250–252 C.
δH [(CD3)2SO] 8.28 (2H, t, J 5.5), 8.03 (2H, s), 7.85 (4H, d, J 8.8), 6.90
4H, d, J 8.7), 3.23 (4H, q, J 6.1), 1.71 (2H, quin, J 6.7). δC [(CD3)2SO]
62.2, 161.9, 150.8, 133.3, 123.3, 117.6, 116.6, 101.4, 37.8, 29.2.
◦
4
2
-Cyano-N-{3-[2-cyano-3-(4-methoxyphenyl)acryloylamino]-
propyl}-3-(4-methoxyphenyl)acrylamide 27
(
1
This was synthesized in the manner described for 5 from 4b and
-methoxybenzaldehyde to yield a yellow solid (88%), mp 190–192 C.
◦
4
δH [(CD3)2SO] 8.31 (2H, t, J 5.2), 8.08 (2H, s), 7.93 (4H, d, J 8.6), 7.10
4H, d, J 8.6), 3.83 (6H, s), 3.27 (4H, q, J 5.8), 1.74 (2H, quin, J 6.3).
δC [(CD3)2SO] 162.5, 161.2, 150.0, 132.4, 124.3, 116.9, 114.7, 102.4,
2
-Cyano-N-{3-[2-cyano-3-(3,4-dihydroxyphenyl)acryloylamino]-
(
propyl}-3-(3,4-dihydroxyphenyl)acrylamide 20
This was synthesized in the manner described for 5 from 4b and
5
5.5, 37.4, 28.7.
◦
3
(
(
,4-dihydroxybenzaldehyde to yield a yellow solid (85%), mp 274 C
[9]
◦
lit. 277 C). δH [(CD3)2SO] 8.24 (2H, br t, J 5.5), 7.92 (2H, s), 7.52
2H, d, J 2.1), 7.26 (2H, dd, J 8.2 and 2.1), 6.85 (2H, d, J 8.2), 3.23 (4H,
2-Cyano-N-{3-[2-cyano-3-(3-hydroxy-4-methoxyphenyl)-
acryloylamino]propyl}-3-(3-hydroxy-4-methoxyphenyl)-
acrylamide 28
q, J 6.0), 1.70 (2H, quin, J 6.7). δC [(CD3)2SO] 161.5, 150.6, 150.5,
1
25.1, 123.2, 117.1, 116.0, 115.8, 100.5, 37.3, 28.8.
This was synthesized in the manner described for 5 from 4b and
3
-hydroxy-4-methoxybenzaldehyde to yield a yellow solid (50%), mp
◦
2
-Cyano-N-{3-[2-cyano-3-(3,4,5-trihydroxyphenyl)acryloylamino]-
194–196 C. δH [(CD ) SO] 8.28 (2H, t, J 5.1), 7.98 (2H, s), 7.53 (2H,
3 2
propyl}-3-(3,4,5-trihydroxyphenyl)acrylamide 21
d, J 1.6), 7.36 (2H, dd, J 8.3 and 1.5), 7.04 (2H, d, J 8.5), 3.84 (6H, s),
3
1
28.7.
.26 (4H, q, J 5.6), 1.73 (2H, quin, J 6.5). δC [(CD3)2SO] 161.4, 151.7,
50.4, 146.7, 124.5, 124.5, 116.5, 114.5, 111.9, 102.1, 102.0, 55.6, 37.4,
This was synthesized in the manner described for 5 from 4b and
,4,5-trihydroxybenzaldehyde to yield a yellow solid (70%), mp >
00 C (lit. > 300 C). δH [(CD3)2SO] 8.18 (2H, t, J 5.5), 7.78 (2H, s),
.99 (4H, s), 3.21 (4H, q, J 6.8), 1.68 (2H, quin, J 6.8). δC [(CD3)2SO]
61.8, 150.7, 145.9, 140.3, 121.2, 117.3, 109.9, 99.5, 38.2, 28.9.
3
3
6
1
◦
[9]
◦
2
-Cyano-N-{3-[2-cyano-3-(4-carboxyphenyl)acryloylamino]-
propyl}-3-(4-carboxyphenyl)acrylamide 29
This was synthesized in the manner described for 5 from 4b and
-carboxybenzaldehyde to yield a white solid (82%), mp 260–264 C.
δH [(CD3)2SO] 8.62 (2H, t, J 5.3), 8.31 (2H, s), 8.10 (4H, d, J 8.0), 7.98
4H, d, J 8.1), 3.23 (4H, q, J 5.7), 1.75 (2H, quin, J 6.6). δC [(CD3)2SO]
66.7, 162.4, 161.0, 149.4, 135.3, 134.3, 129.8, 129.8, 116.1, 115.9,
08.2, 39.4, 26.2.
2
-Cyano-N-{3-[2-cyano-3-(3,4-dihydroxy-5-methoxyphenyl)-
◦
4
acryloylamino]propyl}-3-(3,4-dihydroxy-5-methoxyphenyl)-
acrylamide 22
(
1
1
This was synthesized in the manner described for 5 from 4b and
,4-dihydroxy-5-methoxybenzaldehyde to yield an orange solid (42%),
mp > 300 C. δH [(CD3)2SO] 8.35 (2H, t, J 5.4), 7.95 (2H, s), 7.21 (2H,
d, J 1.9), 7.12 (2H, d, J 1.9), 3.21 (4H, q, J 6.8), 1.71 (2H, quin, J 6.8).
δC [(CD3)2SO] 161.3, 150.6, 147.2, 145.3, 121.0, 117.6, 110.6, 107.6,
3
◦
2-Cyano-N-[4-(2-cyano-3-phenylacryloylamino)butyl]-3-
phenylacrylamide 30
9
8.7, 38.4, 28.9.
Thiswassynthesizedinthemannerdescribedfor 5from 4c andbenz-
aldehyde to yield a yellow solid (85%), mp 188–190 C. δH [(CD3)2SO]
8.51 (2H, t, J 5.6), 7.94 (2H, s), 7.92 (4H, m), 7.56 (6H, m), 3.23 (4H,
◦
3
-(2-Chlorophenyl)-N-{3-[3-(2-chlorophenyl)-
2
-cyanoacryloylamino]propyl}-2-cyanoacrylamide 23
q, J 5.6), 1.53 (4H, br s). δC [(CD ) SO] 160.8, 150.3, 132.2, 131.9,
3 2
1
29.9, 129.2, 116.3, 106.4, 39.3, 26.2.
This was synthesized in the manner described for 5 from 4b and
-chlorobenzaldehyde to yield a yellow solid (80%), mp 172–174 C.
δH [(CD3)2SO] 8.57 (2H, br t, J 5.4), 8.36 (2H, s), 7.98 (2H, dd, J
.1 and 2.1), 7.52 (6H, m), 3.32 (4H, q, J 6.0), 1.77 (2H, quin, J 6.5).
δC [(CD3)2SO] 159.9, 146.8, 134.0, 133.2, 130.3, 130.0, 129.6, 127.7,
15.3, 110.3, 37.6, 28.4.
◦
2
2
-Cyano-N-{4-[2-cyano-3-(3-hydroxyphenyl)acryloylamino]-
butyl}-3-(3-hydroxyphenyl)acrylamide 31
4
This was synthesized in the manner described for 5 from 4c and
◦
1
3-hydroxybenzaldehyde to yield a yellow solid (80%), mp 224–228 C.

Synthesis of Targeted Tyrphostin Libraries with Anticancer Activity
101
δH [(CD3)2SO] 8.46 (2H, t, J 5.5), 8.03 (2H, s), 7.36 (4H, m), 7.32 (2H,
s), 6.96 (2H, m), 3.22 (4H, br s), 1.52 (4H, br s). δC [(CD3)2SO] 160.9,
2-Cyano-N-{4-[2-cyano-3-(3-hydroxy-4-methoxyphenyl)-
acryloylamino]butyl}-3-(3-hydroxy-4-methoxyphenyl)-
acrylamide 39
1
57.7, 150.4, 133.0, 130.2, 121.3, 119.5, 116.3, 115.7, 106.0, 39.3, 26.2.
This was synthesized in the manner described for 5 from 4c and
2
-Cyano-N-{4-[2-cyano-3-(4-hydroxyphenyl)acryloylamino]-
3-hydroxy-4-methoxybenzaldehyde to yield a yellow solid (91%), mp
294–296 C. δH [(CD3)2SO] 8.28 (2H, t, J 5.5), 7.96 (2H, s), 7.52 (2H,
◦
butyl}-3-(4-hydroxyphenyl)acrylamide 32
d, J 1.9), 7.37 (2H, dd, J 8.5 and 1.8), 7.06 (2H, d, J 8.5), 3.84 (6H, s),
This was synthesized in the manner described for 5 from 4c and
-hydroxybenzaldehyde to yield a yellow solid (80%), mp 290–292 C.
δH [(CD3)2SO] 8.28 (2H, t, J 5.6), 8.01 (2H, s), 7.84 (4H, d, J 8.9), 6.90
4H, d, J 8.8), 3.20 (4H, q, J 5.4), 1.49 (4H, br s). δC [(CD3)2SO] 161.7,
61.3, 150.2, 132.7, 122.8, 117.2, 116.1, 101.1, 39.3, 26.3.
3
1
.21 (2H, q, J 5.0), 1.51 (2H, br s). δC [(CD3)2SO] 161.3, 151.6, 150.1,
46.6, 124.5, 116.9, 115.4, 111.9, 102.2, 55.6, 39.3, 26.3.
◦
4
(
1
2-Cyano-N-{4-[2-cyano-3-(4-carboxyphenyl)acryloylamino]-
butyl}-3-(4-carboxyphenyl)acrylamide 40
2
-Cyano-N-{3-[2-cyano-3-(3,4-dihydroxyphenyl)acryloylamino]-
This was synthesized in the manner described for 5 from 4c and
◦
butyl}-3-(3,4-dihydroxyphenyl)acrylamide 33
4-carboxybenzaldehyde to yield a white solid (50%), mp 298–300 C.
δH [(CD3)2SO] 8.59 (2H, t, J 5.3), 8.35 (4H, d, J 8.5), 8.27 (2H, s), 8.10
(4H, d, J 8.6), 3.25 (2H, q, J 4.9), 1.55 (2H, br s). δC [(CD3)2SO] 166.6,
160.5, 149.1, 135.5, 134.0, 129.8, 129.7, 116.0, 108.4, 39.5, 28.6.
This was synthesized in the manner described for 5 from 4c and
◦
3
(
(
,4-dihydroxybenzaldehyde to yield a yellow solid (97%), mp 281 C
[8]
◦
lit. 283 C). δH [(CD3)2SO] 8.25 (2H, t, J 5.5), 7.91 (2H, s), 7.53
2H, d, J 1.9), 7.26 (2H, dd, J 8.3 and 1.9), 6.85 (2H, d, J 8.3), 3.20 (4H,
br s), 1.49 (4H, br s). δC [(CD3)2SO] 161.5, 150.9, 150.4, 145.7, 125.2,
2
-Cyano-N-[5-(2-cyano-3-phenylacryloylamino)pentyl]-3-
1
23.1, 117.2, 115.8, 100.5, 39.3.
phenylacrylamide 41
This was synthesized in the manner described for 5 from 4d and ben-
◦
3
-(2-Chlorophenyl)-N-{4-[3-(2-chlorophenyl)-2-
zaldehyde to yield a yellow solid (51%), mp 164–166 C. δ [(CD ) SO]
H 3 2
cyanoacryloylamino]butyl}-2-cyanoacrylamide 34
8.43 (2H, t, J 5.3), 8.14 (2H, s), 7.91 (4H, m), 7.55 (6H, m), 3.21 (4H,
q, J 6.7), 1.52 (4H, quin, J 7.3), 1.31 (2H, quin, J 6.7). δC [(CD3)2SO]
160.8, 150.2, 132.1, 131.1, 129.9, 129.1, 116.3, 106.5, 39.5, 28.4, 23.7.
This was synthesized in the manner described for 5 from 4c and
-chlorobenzaldehyde to yield a yellow solid (93%), mp 196–198 C.
◦
2
δH [(CD3)2SO] 8.61 (2H, t, J 5.6), 7.98 (2H, s), 7.63 (6H, dd, J 4.2 and
2
1
.0), 7.56 (6H, m), 3.25 (4H, q, J 5.6), 1.54 (4H, br s). δC [(CD3)2SO]
59.9, 146.7, 134.0, 133.1, 130.3, 130.0, 129.6, 127.7, 115.3, 110.5.
2
-Cyano-N-{5-[2-cyano-3-(3-hydroxyphenyl)acryloylamino]-
pentyl}-3-(3-hydroxyphenyl)acrylamide 42
This was synthesized in the manner described for 5 from 4d and
-hydroxybenzaldehyde to yield a yellow solid (92%), mp 230–232 C.
δH [(CD3)2SO] 8.42 (2H, t, J 5.4), 8.02 (2H, s), 7.34 (4H, m), 7.31 (2H,
s), 6.97 (2H, m), 3.19 (4H, q, J 6.0), 1.52 (4H, quin, J 7.0), 1.32 (2H,
3 2
quin, J 6.8). δC [(CD ) SO] 160.9, 157.7, 150.3, 133.0, 130.2, 121.3,
3
-(3-Chlorophenyl)-N-{4-[3-(2-chlorophenyl)-2-
◦
3
cyanoacryloylamino]butyl}-2-cyanoacrylamide 35
This was synthesized in the manner described for 5 from 4c and 3-
◦
chlorobenzaldehyde to yield a yellow solid (73%), mp 202–204 C. δH
(CD3)2SO] 8.48 (2H, t, J 5.5), 8.14 (2H, s), 7.95 (2H, t, J 1.7), 7.85 (2H,
m), 7.60 (4H, m), 3.25 (4H, q, J 5.6), 1.52 (4H, br s). δC [(CD3)2SO]
1
19.5, 116.3, 115.7, 106.1, 39.5, 28.4, 23.6.
[
1
3
60.9, 157.7, 150.4, 133.0, 130.2, 121.3, 119.5, 116.3, 115.7, 106.0,
9.3, 26.2.
2
-Cyano-N-{5-[2-cyano-3-(4-hydroxyphenyl)acryloylamino]-
pentyl}-3-(4-hydroxyphenyl)acrylamide 43
This was synthesized in the manner described for 5 from 4d and
-hydroxybenzaldehyde to yield a yellow solid (98%), mp 242–244 C.
3
-(4-Chlorophenyl)-N-{4-[3-(4-chlorophenyl)-2-
◦
4
cyanoacryloylamino]butyl}-2-cyanoacrylamide 36
δH [(CD3)2SO] 8.24 (2H, t, J 5.5), 7.99 (2H, s) 7.84 (4H, d, J 8.8), 6.88
4H, d, J 8.7), 3.18 (4H, q, J 6.9), 1.48 (4H, quin, J 7.3), 1.28 (2H, quin,
J 6.8). δC [(CD ) SO] 162.0, 161.4, 150.1, 132.7, 122.6, 117.2, 116.2,
(
This was synthesized in the manner described for 5 from 4c and 4-
chlorobenzaldehyde to yield a yellow solid (72%), mp 232–234 C. δH
(CD3)2SO] 8.47 (2H, t, J 5.4), 8.14 (2H, s), 7.93 (4H, d, J 6.9), 7.62
4H, d, J 6.8), 3.23 (4H, q, J 5.5), 1.53 (4H, br s). δC [(CD3)2SO] 160.6,
◦
3 2
1
01.0, 39.5, 28.5, 23.7.
[
(
1
49.0, 136.7, 131.6, 130.7, 129.3, 116.1, 107.0, 39.4, 26.2.
2
-Cyano-N-{3-[2-cyano-3-(3,4-dihydroxyphenyl)acryloylamino]-
pentyl}-3-(3,4-dihydroxyphenyl)acrylamide 44
2
-Cyano-N-{4-[2-cyano-3-(3-methoxyphenyl)acryloylamino]-
This was synthesized in the manner described for 5 from 4d and
butyl}-3-(3-methoxyphenyl)acrylamide 37
◦
3
,4-dihydroxybenzaldehyde to yield a yellow solid (90%), mp 252 C
[
8]
◦
This was synthesized in the manner described for 5 from 4c and
-methoxybenzaldehyde to yield a yellow solid (89%), mp 180–182 C.
δH [(CD3)2SO] 8.46 (2H, br t, J 5.6), 8.12 (2H, s), 7.45 (6H, m), 7.13
2H, m), 3.78 (6H, s), 3.23 (4H, br s), 1.54 (4H, br s). δC [(CD3)2SO]
60.7, 159.3, 150.2, 133.1, 130.2, 122.3, 118.1, 116.3, 114.7, 106.6,
5.2, 39.3, 26.2.
(lit. 248 C). δH [(CD3)2SO] 8.15 (2H, t, J 5.5), 7.85 (2H, s), 7.50
(2H, d, J 2.1), 7.20 (2H, dd, J 8.5 and 2.0), 6.75 (2H, d, J 8.5), 3.16 (4H,
q, J 6.2), 1.50 (4H, quin, J 7.1), 1.28 (2H, quin, J 6.9). δC [(CD3)2SO]
161.9, 153.9, 150.3, 146.3, 126.2, 121.5, 117.7, 115.7, 114.7, 98.4, 39.5,
28.6, 23.7
◦
3
(
1
5
2
-Cyano-N-{5-[2-cyano-3-(3,4-dimethoxyphenyl)acryloylamino]-
2
-Cyano-N-{4-[2-cyano-3-(4-methoxyphenyl)acryloylamino]-
pentyl}-3-(3,4-dimethoxyphenyl)acrylamide 45
butyl}-3-(4-methoxyphenyl)acrylamide 38
This was synthesized in the manner described for 5 from 4d and 3,4-
dimethoxybenzaldehyde to yield a yellow solid (80%), mp > 300 C. δH
◦
This was synthesized in the manner described for 5 from 4c and
-methoxybenzaldehyde to yield a yellow solid (81%), mp 200–202 C.
δH [(CD3)2SO] 8.32 (2H, t, J 5.6), 8.07 (2H, s), 7.94 (4H, d, J 8.9),
.10 (4H, d, J 8.9), 3.83 (6H, s), 3.22 (4H, q, J 5.5), 1.52 (4H, br s).
δC [(CD3)2SO] 162.4, 161.2, 149.8, 132.3, 124.4, 116.9, 114.7, 102.7,
5.5, 39.2, 26.3.
◦
4
[(CD3)2SO] 8.28 (2H, t, J 5.5), 8.05 (2H, s), 7.61 (2H, dd, J 1.9), 7.54
(2H, dd, J 8.5 and 1.8), 7.10 (2H, d, J 8.6), 3.83 (6H, s), 3.78 (6H,
s), 3.20 (4H, q, J 6.1), 1.51 (4H, quin, J 7.1), 1.31 (2H, quin, J 6.8).
δC [(CD3)2SO] 161.1, 152.3, 150.1, 148.5, 125.2, 124.4, 117.1, 112.2,
110.7, 102.6, 55.7, 55.3, 55.3, 39.4, 28.5, 23.6.
7
5

1
02
T. A. Hill et al.
3
-(2-Chlorophenyl)-N-{5-[3-(2-chlorophenyl)-2-
2-Cyano-N-[6-(2-cyano-3-phenylacryloylamino)-
hexyl]-3-phenylacrylamide 53
cyanoacryloylamino]pentyl}-2-cyanoacrylamide 46
This was synthesized in the manner described for 5 from 4d and
-chlorobenzaldehyde to yield a yellow solid (96%), mp 164–166 C.
This was synthesized in the manner described for 5 from 4e and ben-
zaldehyde to yield a yellow solid (72%), mp 166–168 C. δH [(CD3)2SO]
◦
◦
2
δH [(CD3)2SO] 8.56 (2H, t, J 5.5), 8.34 (2H, s), 7.98 (2H, m), 7.54 (6H,
m), 3.23 (4H, q, J 6.1), 1.55 (4H, quin, J 7.0), 1.32 (2H, quin, J 6.7).
δC [(CD3)2SO] 159.9, 146.6, 134.0, 133.1, 130.3, 129.9, 129.6, 127.7,
8.43 (2H, t, J 5.5), 8.14 (2H, s), 7.91 (4H, m), 7.54 (6H, m), 3.21
(4H, quin, J 6.3), 1.50 (4H, quin, J 6.2), 1.31 (4H, br quin, J 6.4).
δC [(CD3)2SO] 160.7, 150.2, 132.1, 131.9, 129.9, 129.1, 116.3, 106.5,
39.6, 28.7, 26.0.
1
15.4, 110.5, 39.6, 28.4, 23.7.
2
-Cyano-N-{6-[2-cyano-3-(3-hydroxyphenyl)acryloylamino]-
3
-(2-Chlorophenyl)-N-{5-[3-(3-chlorophenyl)-2-
hexyl}-3-(3-hydroxyphenyl)acrylamide 54
cyanoacryloylamino]pentyl}-2-cyanoacrylamide 47
This was synthesized in the manner described for 5 from 4e and
-hydroxybenzaldehyde to yield a white solid (70%), mp 234–236 C.
δH [(CD3)2SO] 8.39 (2H, t, J 5.5), 8.02 (2H, s), 7.33 (6H, m), 6.96 (2H,
m), 3.18 (4H, q, J 6.1), 1.50 (4H, br quin, J 6.1), 1.30 (4H, br quin,
J 6.2). δC [(CD3)2SO] 160.8, 157.7, 150.2, 133.0, 130.2, 121.3, 119.5,
This was synthesized in the manner described for 5 from 4d and
-chlorobenzaldehyde to yield a yellow solid (63%), mp 158–160 C.
δH [(CD3)2SO] 8.42 (2H, t, J 5.4), 8.12 (2H, s), 7.94 (2H, t, J 1.6),
.85 (2H, m), 7.58 (2H, m), 3.20 (4H, q, J 6.7), 1.52 (4H, quin, J 7.2),
.32 (2H, quin, J 5.2). δC [(CD3)2SO] 160.4, 148.6, 133.9, 133.7, 131.6,
30.9, 129.2, 128.3, 128.2, 115.9, 108.1, 39.2, 28.3, 23.6
◦
3
◦
3
7
1
1
1
16.3, 115.7, 106.1, 39.6, 28.7, 26.0.
2
-Cyano-N-{6-[2-cyano-3-(4-hydroxyphenyl)acryloylamino]-
3
-(4-Chlorophenyl)-N-{5-[3-(4-chlorophenyl)-2-
hexyl}-3-(4-hydroxyphenyl)acrylamide 55
cyanoacryloylamino]pentyl}-2-cyanoacrylamide 48
This was synthesized in the manner described for 5 from 4e and
◦
This was synthesized in the manner described for 5 from 4d and
-chlorobenzaldehyde to yield a yellow solid (84%), mp 160–162 C.
δH [(CD3)2SO] 8.45 (2H, t, J 5.3), 8.12 (2H, s), 7.90 (4H, d, J 6.9), 7.60
4H, d, J 8.6), 3.20 (4H, q, J 6.0), 1.51 (4H, quin, J 7.1), 1.32 (2H, quin,
J 6.7). δC [(CD3)2SO] 160.6, 148.8, 136.7, 131.5, 130.7, 129.2, 116.1,
4-hydroxybenzaldehyde to yield a yellow solid (65%), mp 272–276 C.
δH [(CD3)2SO] 8.21 (2H, t, J 5.4), 8.00 (2H, s), 7.84 (4H, d, J 8.7), 6.90
(4H, d, J 8.5), 3.18 (4H, q, J 6.2), 1.48 (4H, br quin, J 6.0), 1.29 (4H, br
quin, J 6.3). δC [(CD3)2SO] 161.7, 161.3, 150.1, 132.7, 132.7, 122.8,
117.2, 116.1, 101.2, 39.5, 28.8, 26.0.
◦
4
(
1
07.0, 39.5, 28.4, 23.6.
2
-Cyano-N-{3-[2-cyano-3-(3,4-dihydroxyphenyl)acryloylamino]-
hexyl}-3-(3,4-dihydroxyphenyl)acrylamide 56
2
-Cyano-N-{5-[2-cyano-3-(3-methoxyphenyl)acryloylamino]-
This was synthesized in the manner described for 5 from 4e and 3,4-
pentyl}-3-(3-methoxyphenyl)acrylamide 49
◦
[8]
dihydroxybenzaldehyde to yield a yellow solid (89%), mp 263 C (lit.
This was synthesized in the manner described for 5 from 4d and
-methoxybenzaldehyde to yield a yellow solid (96%), mp 124–126 C.
δH [(CD3)2SO] 8.43 (2H, t, J 5.6), 8.10 (2H, s), 7.47 (6H, m), 7.13 (2H,
m), 3.79 (6H, s) 3.21 (4H, q, J 6.7), 1.51 (4H, quin, J 7.1), 1.32 (2H,
quin, J 6.7). δH [(CD3)2SO] 160.7, 159.3, 150.1, 133.1, 130.2, 122.3,
◦
2
2
1
1
60 C). δH [(CD3)2SO] 8.18 (2H, t, J 5.5), 7.89 (2H, s), 7.51 (2H, d, J
◦
3
.0), 7.24 (2H, dd, J 2.0 and 8.3), 6.83 (2H, d, J 8.3), 3.17 (4H, q, J 6.1),
.47 (4H, quin, J 6.1), 1.28 (4H, br s). δC [(CD3)2SO] 161.5, 151.2,
50.3, 145.7, 125.2, 122.9, 117.2, 115.8, 115.7, 100.4, 39.5, 28.8, 26.0.
1
18.0, 116.3, 114.7, 106.7, 55.2, 39.5, 28.4, 23.6.
3
-(2-Chlorophenyl)-N-{6-[3-(2-chlorophenyl)-2-
cyanoacryloylamino]hexyl}-2-cyanoacrylamide 57
2
-Cyano-N-{5-[2-cyano-3-(4-methoxyphenyl)acryloylamino]-
This was synthesized in the manner described for 5 from 4e and
-chlorobenzaldehyde to yield a yellow solid (91%), mp 160–162 C.
pentyl}-3-(4-methoxyphenyl)acrylamide 50
◦
2
δH [(CD3)2SO] 8.56 (2H, t, J 5.5), 8.33 (2H, s), 7.98 (2H, m), 7.56 (6H,
m), 3.22 (4H, q, J 6.3), 1.52 (4H, quin, J 5.9), 1.32 (4H, br quin, J 6.2).
δC [(CD3)2SO] 159.9, 146.6, 134.0, 133.1, 130.4, 130.0, 129.6, 127.7,
This was synthesized in the manner described for 5 from 4d and
-methoxybenzaldehyde to yield a yellow solid (87%), mp 132–134 C.
δH [(CD3)2SO] 8.30 (2H, t, J 5.6), 8.06 (2H, s), 7.94 (4H, d, J 8.9), 7.08
4H, d, J 8.9), 3.83 (6H, s), 3.20 (4H, q, J 6.0), 1.50 (4H, quin, J 7.1),
◦
4
1
15.3, 110.6, 39.7, 28.6, 26.0.
(
1
1
.30 (4H, quin, J 5.6). δC [(CD3)2SO] 162.4, 161.1, 149.7, 132.3, 124.4,
16.9, 114.7, 102.7, 55.5, 39.5, 28.5, 23.6.
3
-(2-Chlorophenyl)-N-{6-[3-(2-chlorophenyl)-2-
cyanoacryloylamino]hexyl}-2-cyanoacrylamide 58
This was synthesized in the manner described for 5 from 4e and
-chlorobenzaldehyde to yield a yellow solid (85%), mp 192–194 C.
δH [(CD3)2SO] 8.44 (2H, t, J 5.5), 8.13 (2H, s), 7.95 (2H, t, J 1.8), 7.85
2H, m), 7.61 (2H, m), 3.19 (4H, q, J 6.8), 1.51 (4H, quin, J 6.5), 1.30
4H, br quin, J 6.4). δC [(CD3)2SO] 160.4, 148.6, 134.0, 133.7, 131.6,
2
-Cyano-N-{5-[2-cyano-3-(3-hydroxy-4-methoxyphenyl)-
◦
3
acryloylamino]pentyl}-3-(3-hydroxy-4-methoxyphenyl)-
acrylamide 51
(
(
1
This was synthesized in the manner described for 5 from 4d
and 3-hydroxy-4-methoxybenzaldehyde to yield a yellow solid (92%),
mp 196–198 C. δH [(CD3)2SO] 8.27 (2H, t, J 5.5), 7.94 (2H, s), 7.51
31.0, 129.2, 128.3, 115.9, 108.2, 39.2, 28.61, 25.9.
◦
(
2H, d, J 2.2), 7.36 (2H, dd, J 8.6 and 2.2), 7.05 (2H, d, J 8.6), 3.84 (6H,
3-(4-Chlorophenyl)-N-{6-[3-(4-chlorophenyl)-2-
cyanoacryloylamino]hexyl}-2-cyanoacrylamide 59
s), 3.19 (2H, q, J 6.6), 1.50 (2H, quin, J 7.0), 1.29 (2H, quin, J 4.7).
δC [(CD3)2SO] 161.3, 151.6, 150.0, 146.6, 124.5, 116.9, 115.4, 111.9,
1
This was synthesized in the manner described for 5 from 4e and
-chlorobenzaldehyde to yield a yellow solid (72%), mp 198–200 C.
02.3, 55.6, 39.5, 28.5, 23.7.
◦
4
δH [(CD3)2SO] 8.45 (2H, t, J 5.3), 8.12 (2H, s), 7.92 (2H, d, J 6.9), 7.62
(2H, d, J 6.9), 3.20 (4H, quin, J 6.2), 1.50 (4H, quin, J 6.0), 1.31 (4H, br
2
-Cyano-N-{5-[2-cyano-3-(4-carboxyphenyl)acryloylamino]-
pentyl}-3-(4-carboxyphenyl)acrylamide 52
quin, J 6.0). δC [(CD ) SO] 160.5, 148.9, 136.7, 131.5, 130.8, 129.2,
3 2
1
16.0, 107.0, 39.6, 28.6, 26.0.
This was synthesized in the manner described for 5 from 4d and
-carboxybenzaldehyde to yield a yellow solid (74%), mp 226–228 C.
δH [(CD3)2SO] 8.52 (2H, t, J 5.1), 8.18 (2H, s), 8.03 (4H, d, J 8.9), 7.98
4H, d, J 8.9), 3.21 (4H, q, J 6.4), 1.54 (4H, quin, J 6.9), 1.32 (2H, quin,
J 4.9). δC [(CD3)2SO] 166.6, 160.5, 149.1, 135.5, 134.0, 129.8, 129.7,
15.9, 108.4, 39.5, 28.4, 23.6.
◦
4
2
-Cyano-N-{6-[2-cyano-3-(3-methoxyphenyl)acryloylamino]-
hexyl}-3-(3-methoxyphenyl)acrylamide 60
(
This was synthesized in the manner described for 5 from 4e and
◦
1
3-methoxybenzaldehyde to yield a yellow solid (77%), mp 136–138 C.

Synthesis of Targeted Tyrphostin Libraries with Anticancer Activity
103
δH [(CD3)2SO] 8.42 (2H, t, J 5.5), 8.12 (2H, s), 7.46 (6H, m), 7.14 (2H,
m), 3.80 (6H, s), 3.22 (4H, q, J 6.0), 1.51 (4H, br quin, J 6.1), 1.31 (4H,
br quin, J 6.1). δC [(CD3)2SO] 160.7, 159.3, 150.1, 133.1, 130.2, 122.3,
References
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18.0, 116.3, 114.7, 106.7, 55.2, 39.6, 28.7, 26.0.
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-Cyano-N-{6-[2-cyano-3-(4-methoxyphenyl)acryloylamino]-
hexyl}-3-(4-methoxyphenyl)acrylamide 61
This was synthesized in the manner described for 5 from 4e and
-methoxybenzaldehyde to yield a yellow solid (98%), mp 182–184 C.
δH [(CD3)2SO] 8.29 (2H, t, J 5.5), 8.06 (2H, s), 7.93 (2H, d, J 7.2), 7.11
2H, d, J 7.2), 3.84 (6H, s), 3.17 (4H, q, J 5.7), 1.49 (4H, br quin, J 5.9),
.30 (4H, br quin, J 6.2). δC [(CD3)2SO] 162.4, 161.0, 149.7, 132.23,
[
[
[
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[
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4
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-Cyano-N-{6-[2-cyano-3-(3-hydroxy-4-methoxyphenyl)-
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This was synthesized in the manner described for 5 from 4e and
-hydroxy-4-methoxybenzaldehyde to yield a yellow solid (99%), mp
26–228 C. δH [(CD3)2SO] 8.26 (2H, t, J 5.6), 7.95 (2H, s), 7.52 (2H,
d, J 1.9), 7.36 (2H, dd, J 8.2 and 1.9), 7.06 (2H, d, J 8.2), 3.84 (6H, s),
.18 (4H, q, J 5.8), 1.49 (4H, br quin, J 6.0), 1.29 (4H, br quin, J 5.9).
δC [(CD3)2SO] 161.3, 151.6, 150.1, 146.7, 124.6, 124.5, 116.9, 115.4,
3
3
2
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CO;2-N
hexyl}-3-(4-carboxyphenyl)acrylamide 63
This was synthesized in the manner described for 5 from 4e and
-carboxybenzaldehyde to yield a yellow solid (67%), mp 288–292 C.
δH [(CD3)2SO] 8.53 (2H, t, J 5.1), 8.18 (2H, s), 8.07 (4H, d, J 8.3),
.98 (4H, d, J 8.3), 3.20 (4H, q, J 5.8), 1.51 (4H, br s), 1.31 (4H, br s).
δC [(CD3)2SO] 166.6, 160.5, 149.1, 135.5, 134.0, 129.8, 129.7, 116.0,
08.4, 39.5, 28.6, 26.0.
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◦
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0
0480-X
7
1
Acknowledgments
We acknowledge financial support from the University of
Newcastle and the NHMRC, Australia.