In vitro activity of novel dual action MDR anthranilamide modulators with inhibitory activity at CYP-450

Article abstract of DOI:10.1016/j.bmc.2006.07.055

Synthesis and in vitro cytotoxicity assays of new anthranilamide MDR modulators have been performed to assess their inhibition potency of the P-glycoprotein (P-gp) transporter. The aromatic spacer group between nitrogen atoms (N¹ and N²

Full text of DOI:10.1016/j.bmc.2006.07.055

Bioorganic & Medicinal Chemistry 14 (2006) 7972–7987
In vitro activity of novel dual action MDR anthranilamide
modulators with inhibitory activity at CYP-450
c
a
d
Philippe Labrie,a,b, Shawn. P. Maddaford, Jacques Lacroix, Concettina Catalano,
*
d
c
a
´
David K. H. Lee, Suman Rakhit and Rene C. Gaudreault
aInstituts de Biomateriaux et des Biotechnologies, Hopital St-Franc¸ois d’Assise, Laval University, Que., Canada G1L 3L5
bFaculty of Pharmacy, Laval University, Quebec City, Que., Canada, G1K 7P4
cNeurAxon Inc. MaRS Centre, South Tower, 101 College Street, Suite 200, Room 230, Toronto, Ont., Canada M5G 1L7
dNoAb BioDiscoveries Inc., 2820 Argentia Road, Unit 8, Mississauga, Ont., Canada L5N 8G4
Received 28 February 2006; revised 11 July 2006; accepted 26 July 2006
Available online 10 August 2006
Abstract—Synthesis and in vitro cytotoxicity assays of new anthranilamide MDR modulators have been performed to assess their
inhibition potency of the P-glycoprotein (P-gp) transporter. The aromatic spacer group between nitrogen atoms (N1 and N2) in the
known inhibitor XR9576 was replaced with a flexible alkyl chain of 2 to 6 carbon atoms in length. 6,7-Dimethoxy-1,2,3,4-tetrahy-
droisoquinoline and their open-chain N-methylhomoveratrylamine counterparts were shown to be potent P-gp inhibitors. The max-
imal inhibition was obtained when using an ethyl or propyl spacer. Several compounds were more potent than verapamil and
intrinsically less cytotoxic than XR9576. In addition, in vitro metabolism studies of 23a with a subset of human CYP-450 isoforms
revealed that, unlike XR9576, 23a inhibited CYP3A4, an enzyme that colocalizes with P-gp in the intestine and contributes to tumor
cell chemoresistance by enhancing the biodisposition of anticancer drugs such as paclitaxel toward metabolism. In this context, 22a
might be a suitable candidate for further drug development.
ꢀ 2006 Elsevier Ltd. All rights reserved.
1. Introduction
exhibiting excretory functions such as liver, kidney,
and intestine.4 Expression of P-gp in such tissues results
in reduced drug absorption from the gastrointestinal
tract and enhanced drug elimination into bile and urine.
Moreover, expression of P-gp in endothelial cell mem-
branes of the blood–brain barrier prevents the diffusion
of drugs into the central nervous system. Therefore,
P-gp plays a key role in tumor chemoresistance via sig-
nificant alterations of anticancer drugs’ pharmacokinet-
ics (e.g., decrease of bioavailability) and biodistribution.
Furthermore, P-gp can act synergistically with cyto-
chrome P450 (CYP) 3A4 to enhance drug metabolism
and elimination.5
A major limitation to the successful chemotherapeutic
treatment of cancer is the resistance, both natural and
acquired, of tumor cells to cytotoxic drugs.1 The overex-
pression of drug transport proteins is a major mecha-
nism for this multiple drug resistance (MDR).2
P-glycoprotein (P-gp) is a member of the ATP-binding
cassette (ABC) family of membrane transporters that
transfer substrates out of cells in an unidirectional fash-
ion.3 In tumor cells overexpressing P-gp, this results in
reduced intracellular drug concentrations which in turn
decrease the efficacy of a broad spectrum of antitumor
drugs including 5-12-anthracyclinediones (e.g., dauno-
rubicin), vinca alkaloids (e.g., vincristine), podophyllo-
toxins (e.g., etoposide), and taxus alkaloids (e.g., taxol).
The inhibition of P-gp and other proteins related to
MDR is of utmost importance in modern cancer chemo-
therapy. It is hoped that P-gp inhibitors could restore
the therapeutic activity of anticancer agents. In that
context, the MDR phenotype can be reversed in P-gp
-positive cells by a large number of structurally hetero-
geneous molecules including notably calcium-channel
P-gp is increasingly recognized as a key determinant in
drug disposition given its expression in normal tissues
Keywords: Multidrug resistance; P-glycoprotein modulators; Anthra-
nilamide; Cytochrome P-450.
blockers
(e.g.,
verapamil)6
immunomodulators
(e.g., cyclosporin A, FK506)7 cardiac antiarrhythmic
agents (e.g., quinidine)8 hypotensives (e.g., reserpine,9
*
Corresponding author. Tel.: +1 418 656 2131x7989; fax: +1 418 525
4272; e-mail: plabrie16@hotmail.com
0968-0896/$ - see front matter ꢀ 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2006.07.055

P. Labrie et al. / Bioorg. Med. Chem. 14 (2006) 7972–7987
7973
piperazine analogues10), dihydropyridines (e.g., nicardi-
pine)11 and many others.12 Consequently, clinical trials
using therapeutically relevant drugs such as verapamil,13
tamoxifen,14 progesterone15 and cyclosporin A16 have
been conducted albeit with very limited successes owing
to their intrinsic toxicity and inability to achieve phar-
macologically effective plasma levels. A second genera-
tion of P-gp modulators such as cyclosporin D
analogues (e.g., PSC 833),17 dexverapamil, biricodar
(VX-710), GF120918,18 and MS209 exhibited improved
P-gp selectivity but remained unselective toward other
ABC transporters leading to deleterious drug-drug inter-
actions. More recently, a third generation of inhibitors
such as XR9576 (tariquidar),19,20 cyclopropyldibenzos-
uberane derivatives (LY335979),21 and diarylimidazoles
(OC144-093)22 has been developed. The anthranilamide
derivative XR9576 is able to reverse primary doxorubi-
cin, vinblastine, and paclitaxel resistance in advanced
breast cancer. Unfortunately, a recent phase II clinical
trial on breast carcinoma and a phase III on lung carci-
noma assessing the efficacy of tariquidar in combination
with vinorelbine versus vinorelbine alone were discon-
tinued due to a significant proportion of adverse events
in the tariquidar cohort. In addition a third clinical trial
testing paclitaxel/carboplatin/tariquidar in combination
was also terminated due to poor response rates and tox-
icity.23 Therefore, there are unmet needs for newer P-gp
inhibitors with reduced intrinsic toxicity.
N1 and N2 on the in vitro activity (Fig. 1). To this
end, we replaced the aromatic linker by alkyl chains of
increasing length. In addition, we synthesized
conformationally flexible open A-ring analogues corre-
sponding to the N-methylhomoveratrylamine deriva-
tives (Fig. 2). All compounds were compared to
verapamil (VRP) and XR9576. Herein we describe our
efforts to identify new anthranilamide-based P-gp
inhibitors.
2. Chemistry
The preparation of the target compounds was
achieved from commercially available veratrylamine
1a (Scheme 1) and dimethoxytetrahydroisoquinoline
1b (Scheme 2). The nucleophilic substitution of the
secondary amines 1a or 1b and the chloronitriles 2–6
in the presence of base generates the corresponding
aminonitriles 7a–11a and 7b–11b in good yields.
Hydrogenation of 7a–11a or 7b–11b with Raney nickel
at 40 psi produced the amines 12a–16a and 12b–16b,
respectively. The synthesis of 17a–21a or 17b–21b
was carried out by the acylation of isatoic anhydride
with amines 12a–16a or 12b–16b in acetonitrile. After-
wards, anilines 17a–21a or 17b–21b were reacted with
3-quinoloyl chloride in the presence of Dowex 1X8
basic resin to produce compounds 22a–26a.
We report here the synthesis of new P-gp inhibitors
based on anthranilamide derivatives such as XR9576
(Fig. 1). As aforementioned, it is known that anthranila-
mides abrogate cell chemoresistance at low nanomolar
levels. While the anthranilamide nucleus tolerates a vari-
ety of substitutions19 with modest reduction of potency,
due to the potential toxicity of the anilide core,20 we
decided to investigate the replacement of the anilide
linker and investigate the effect of the distance between
An alternative synthetic approach to the synthesis of
new anthranilamides such as compound 24b was investi-
gated (Scheme 2). The synthetic pathway of 24b was
identical except that the acylation reaction was per-
formed using 2-nitrobenzoyl chloride instead of isatoic
anhydride to produce the corresponding nitro derivative
19. The nitro group was reduced by zinc metal in pres-
ence of ammonium chloride in refluxing methanol to
give the aniline 19b in 72% yields. Finally, the synthesis
of 24b was completed as by acylation of 19b by 3-quino-
loyl chloride using the basic resin Dowex 1X8 as an acid
scavenger.
3. In vitro biological evaluation
The P-gp inhibition potency of compounds 22a–26a and
22b–26b was assessed using CEM/VLB500 human leuke-
mia cells. The assay is based on the ability of the drug to
revert the chemoresistance of these cells to daunorubicin
Figure 1. Structure of XR9576.
Figure 2. Molecular structures of target molecules.

7974
P. Labrie et al. / Bioorg. Med. Chem. 14 (2006) 7972–7987
Scheme 1. Preparation of the N-methylhomoveratrylamine derivatives 22a–26a. Reagents and conditions: (a) Method A. K2CO3 or Na2CO3, NaI,
t-BuOH, reflux 24 h. Method B. Acetonitrile or DMF, K2CO3, reflux 24 h; (b) H2, Raney Ni, 40 psi, 1.4 M NaOH/EtOH, 12 h; (c) isatoic anhydride,
acetonitrile, reflux, 18 h; (d) 3-quinoline acid chloride, CH2Cl2, 0–25 ꢁC, Dowex 1X8 resin (100–200 mesh), 30 h.
Scheme 2. Preparation of the tetrahydroisoquinoline derivatives 22b–26b. Reagents and conditions: (a) Method A. K2CO3 or Na2CO3, NaI, t-BuOH,
reflux 24 h; Method B. Acetonitrile or DMF, K2CO3, reflux 24 h; (b) H2, Raney Ni, 40 psi, 1.4 M NaOH/EtOH, 12 h; (c) i—isatoic anhydride,
acetonitrile, reflux, 18 h or (d) 2-nitrobenzoylchloride, DOWEX 1X8 resin (100–200 mesh), ethyl acetate (e) NH4Cl, Zn, MeOH, reflux, 90 min.
(f) 3-quinoline acid chloride, CH2Cl2, 0–25 ꢁC, Dowex resin 1X8 (100–200 mesh), 30 h.
(DNR) and vinblastine (VBL). These cells exhibit a high
level of P-gp overexpression. As depicted in Figure 3,
CEM/VLB500 cells were more than 5500-fold resistant
to VLB than their wild-type counterpart (CEM).
(0–15 lM) of the inhibitor either alone or in the presence
of vinblastine (100 nM) or daunorubicin (100 nM), which
are both P-gp substrates. After three days of treatment,
the GI50 of the inhibitor (Table 1) (concentration of inhib-
itor necessary to kill 50% of the cells) was determined
using the resazurin assay.24 Also, a ‘difference score’
(EC50) based on the difference between the number of cells
The ability to reverse MDR in resistant cells was deter-
mined by treating the cells with escalating concentrations

P. Labrie et al. / Bioorg. Med. Chem. 14 (2006) 7972–7987
7975
Table 2. Inhibition activity of new anthranilamide P-gp inhibitors on
CEM/VBL500 cell incubated in the presence of vinblastine or
daunorubicin
CEM_DNR
CEM/VLB500_DNR
CEM_VLB
na
CEM/VLB500–VLB
EC50 (nM)
CEM/VLB500–DNR
EC50 (nM)e
100
90
80
70
60
50
40
30
20
10
0
Compound
CEM/VLB500_VLB
22a
23a
1
270 84b
490 90b
338 27
293 92
352 121
843 260
539 32
1680 200
652 152
860 24
741 171
769 376
—
2
24a
25a
3
781 177b
1345 113d
904 384c
899 127d
500 147d
945 346d
1274 27d
1084 255d
68 40c
4
26a
22b
5
1
23b
24b
2
3
25b
26b
4
5
XR9576
VRP
—
—
1041 443b
799 59
0,1
1
10
100
1000 10000 10000
0
a Number of linker carbon atoms between the nitrogen atoms is n + 1.
b Average of four experiments performed in quadruplicate.
c Average of five experiments performed in quadruplicate.
d Average of two experiments performed in quadruplicate.
e Average of three experiments performed in quadruplicate.
Concentration of DNR or VLB (nM)
Figure 3. Comparison of cytotoxic activity of VLB and DNR against
CEM and CEM/VLB500 MDR cells.
Table 3. In vitro cytochrome-P450 inhibition assays of 22a and
XR9576
Table 1. Cytotoxic activity of new anthranilamides on CEM/VBL500
cells
Reference
Reference XR9576 22a
IC50
Compound
na
CEM/VLB500 GI50 (nM)
IC50
(lM)
IC50
(lM)
22a
23a
1
27707 15909c
29177 13401c
19063 7858c
17845 10300b
10201 2147d
42298 4588b
29339 11787b
26353 4588b
17210 4952b
17060 9197b
11350 3100b
>15,000
(lM)
2
CYP1A2
CYP2A6
CYP2B6
CYP2C8
CYP2C9
CYP2C19
CYP2D6
CYP2E1
Furafylline
Tranylcypromine
4.98
0.98
27.2
>100
>100
45.3
7.1
>100
>100
>100
5.82
24a
25a
3
4
Tranylcypromine 10.3
Quercetin
26a
22b
5
1.56
0.56
1
Sulfaphenazole
Tranylcypromine
Quinidine
2.08
14.7
23b
24b
2
8.74
0.0074
>10
100
3
>100
>100
1.62
25b
26b
4
DDTC
17.1
>100
>100
>100
5
CYP3A4/BFC Ketoconazole
CYP3A4/BQ Ketoconazole
0.096
0.26
XR9576
VRP
—
—
7.75
BFC, 7-benzyloxy-4-trifluoromethylcoumarin; BQ, 7-benzyloxyquin-
oline; DDTC, diethyldithiocarbamic acid.
a Number of linker carbon atoms between the nitrogen atoms is n + 1.
b Average of six experiments performed in quadruplicate.
c Average of nine experiments performed in quadruplicate.
d Average of eight experiments performed in quadruplicate.
inhibit their catalytic activity. The assays are based on
the capacity of a given drug to inhibit or to increase
the metabolism of a specific substrate of the CYP yield-
ing in a fluorescent metabolite. In that context, micro-
that survived in presence of the modulator alone (cytotox-
icity of the inhibitor) minus the number of cells that sur-
vived in presence of the modulator and the cytotoxic
agent25 was calculated (Table 2).
somes from an insect cell transfected with
a
baculovirus expressing individual human CYP450 sub-
types were prepared.27 The drug was incubated with
the microsomes expressing a specific CYP. Afterwards,
the CYP substrate was added and the kinetics of the
substrate metabolism was established using fluores-
cence. Of interest, inhibition constants are substrate
dependent for CYP3A4 and to avoid potential misinter-
pretation of the results two substrates (7-benzyloxy-4-
trifluoromethylcoumarin and 7-benzyloxyquinoline)
were used. The IC50 are listed in Table 3.
4. In vitro CYP450 inhibition studies
Many drug–drug interactions are initiated by the metab-
olism of drugs by cytochrome P-450 (CYP). Eleven
CYPs metabolizing xenobiotics namely CYP1A2,
CYP2A6, CYP2B6, CYP2C8/9/18/19, CYP2D6,
CYP2E1, and CYP 3A4/5 are expressed in a typical hu-
man liver. However, CYP1A2, CYP2C8, CYP2C9,
CYP2C19, CYP2D6, and CYP3A4 appear to be in-
volved in the metabolism, and the drug–drug interac-
tions of commonly used drugs.26 Our hit P-gp
inhibitor 22a along with XR9576 were examined in a
CYP450 inhibition assay to evaluate their ability to
5. Results and discussion
Our results have shown that compounds 22a–26a and
22b–26b exhibit potent MDR reversal activity. In addi-
tion, most of our compounds were at least as potent as

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P. Labrie et al. / Bioorg. Med. Chem. 14 (2006) 7972–7987
VRP as P-gp inhibitor. Three compounds 22a, 23a,
and23b were more active than verapamil. Compound
22a was 4-fold more potent than verapamil when tested
in combination with VLB and twice as potent when
combined with DNR. In addition 23a and 23b were
twice as potent as verapamil with VLB. In combination
with DNR, 23a was about 3-fold more potent than
verapamil and 23b was equipotent to verapamil. The
selection of a ‘hit’ or a ‘lead compound’ cannot be based
solely on its ability to inhibit the activity of its target.
Therefore, we have evaluated the cytotoxicity of the
drug itself and its ability to be involved in drug–drug
interactions through CYP inhibition or induction. These
preliminary experiments aimed to determine if a drug
candidate will potentially trigger deleterious effects in
later animal experiments. As shown in Figure 4 and
Table 1, our compounds exhibited minimal cytotoxicity
at concentrations up to 10 lM while the evaluated
XR9576 was significantly toxic at this concentration.
At concentrations of up to 10 lM, 22a is non-toxic
alone, but killed cells in combination with vinblastine,
while verapamil has half of the activity of 22a and is tox-
ic by itself.
tetrahydroisoquinoline series, the most active compound
bears a 3-carbon atom spacer (n = 2, 23b). Similar trends
are observed for both vinblastine and daunorubicin.
Studies of the effect of 22a and XR9576 on human CYP
were also conducted using several human cytochrome
P-450 enzymes, notably CYP1A2, CYP2A6, CYP2B6,
CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1 or
CYP3A4, to determine the ability of these compounds
to inhibit the metabolism of cytotoxic drugs thereby
increasing their accumulation in cells. The results in
Table 3 show that 22a has a CYP inhibition profile sig-
nificantly different from the one exhibited by XR9576.
Indeed, 22a inhibited CYP2C8 (5.82 lM), CYP2C9
(2.08 lM), CYP2C19 (14.7 lM), and CYP3A4 (1.62
for BFC and 7.75 lM for BQ) at lM level, while
XR9576 inhibited CYP1A2, CYP2C8, and CYP2C9 at
27.2, 45.3 and 7.1 lM, respectively. These differences
suggest that 22a may have a greater impact on the
metabolism of cytotoxic drugs such as paclitaxel, dau-
norubicin, vinblastine, and etoposide in comparison to
XR9576. CYP3A4 accounts for approximately 25% of
the total hepatic CYP enzymes and are responsible for
the metabolism of the majority of cytotoxic drugs.28
Fortuitously, P-gp shares a broad substrate overlap with
CPY3A4 and is colocalized in the small intestine, which
can also limit the gastrointestinal absorption of several
drugs.29 The apparent colocalization of CYP3A4 and
P-gp could act synergistically to enhance drug metaboli-
zation via a recycling process in the gut. Therefore, an
inhibitor of P-gp that is also an inhibitor of CYP3A4
might provide a therapeutical advantage by enhancing
the bioavailability of the anticancer drugs through a
decrease of its excretion and a decrease of its biotrans-
formation. Several investigators reported that poor bio-
availability of paclitaxel might be due to CYP and P-gp
present in the gut wall.30–32 To show the importance
CYP450 inhibition in improving the absorption of pac-
litaxel, administration of known P-gp inhibitor
LY335979 was shown to reduce paclitaxel clearance by
approximately 25% in a recent study.33 This improve-
ment, although modest, is likely due to its weak inhibi-
tion of CYP3A4, as LY335979 is about 60-fold more
selective for P-gp than for CYP3A4.34 In our studies,
22a inhibited both CYP3A4 and P-gp, while XR9576
inhibited only P-gp. Both 22a and XR9576 may increase
the accumulation of drugs in the cell, but 22a may also
enhance their bioavailability due to its inhibitory effect
on CYP3A4 at lM levels.
Some general trends seem to emerge from the new com-
pounds in terms of the effect of the modifications on the
A-ring versus the P-gp inhibition. Indeed, the series
containing the N-methylhomoveratrylamine group
(22a–26a) are generally more active compared to the
tetrahydroisoquinoline series (22b–26b). These results
suggest that the flexibility of the linker between N1
and N2 might be important to the inhibitory activity.
In addition, the distance between N1 and N2 may also
influence the activity of the drug; the optimal activity
being observed with an ethyl chain (n = 1). In the N-meth-
ylhomoveratrylamine series 22a–26a, the activity steadily
decreases as the number of carbon atoms in the linking
arm increases to up to 5 atoms. The activity of 26a
with 6carbonatoms distance betweenN1 and N2 is similar
to the 4-carbon atom chain of compound 24a. In the
There is increasing evidence suggesting that the presence
of intratumoral enzymes, such as CYP450, may play an
important role in the development of cancer chemoresis-
tance.35 Paclitaxel is a taxus derivative metabolized by
CYP3A4. In a recent study on 23 patients having breast
cancers, the analysis of CYP3A4 mRNA levels in tumor
biopsies before docetaxel treatment revealed that pa-
tients with low CYP3A4 mRNA levels had significantly
higher response rates compared to those with high
CYP3A4 mRNA levels. This suggests that overexpres-
sion of CYP3A4 may provide a protective survival
advantage to cancer cells by the inactivation of cytotoxic
agents.
Figure 4. Comparison of the chemoresistance of 22a, 23a, 24a,
verapamil, and XR9576 in vinblastine treated CEM/VLB500 MDR
cells. Difference score = (percent survival of cells treated with test agent
alone) À (percent survival of cells treated with test compound and
100 nM vinblastine). The maximum variation of these data points was
10%. See Section 7 for details.

P. Labrie et al. / Bioorg. Med. Chem. 14 (2006) 7972–7987
7977
6. Conclusions
plicate. The maximum variation of these data point was
10%. The difference score was plotted in function of
the concentration of the modulator to determine an
EC50 that represents the concentration that is necessary
to revert chemoresistance by 50%.37
In this work, we have demonstrated that the N-methylho-
moveratrylamine series 22a–26a exhibits a more potent
P-gp inhibition than the tetrahydroisoquinoline series
22b–26b. These results suggest that the flexibility of the
linker arm between N1 and N2 might be an important fac-
tor modulating the potency. We have also shown that a
two- or three-carbon atom linker between N1 and N2
seems optimal for P-gp inhibition. Compound 22a was
the most potent of the series of molecules that were pre-
pared; it was four times more active than verapamil, but
four times less active than XR9576. Although less active
than XR9576 as P-gp inhibitor, compound 22a has been
shown to be less intrinsically cytotoxic than VRP and
XR9576. In addition, 22a has a different CYP inhibition
profile compared to XR9576. In contrast to XR9576,
22a inhibited CYP 3A4, which is known to colocalize with
P-gp and that is a major impediment to chemotherapy.
Compound 22a might be a therapeutically interesting
molecule that is able to improve the bioavailability and
half-life of a number of anticancer drugs such as paclitaxel
through inhibition of P-gp and bioinactivation through
metabolism, respectively.
7.3. CYP450 inhibition assay
A general description of the assays follows, with enzyme-
specific parameters listed separately below for each CYP
subtype. The CYP450 inhibition assays were conducted
using microsomes (Supersomesꢂ, BD GENTEST,
Woburn, MA) prepared from insect cells, each expressing
a specific CYP subtype (CYP1A2, CYP2C9, CYP2C19,
CYP2D6, or CYP3A4) expressed from the corresponding
human CYP cDNA using a baculovirus expression vec-
tor. The microsomes also incorporate supplemental
cDNA-expressed human reductase and/or cytochrome
b5, as these enzymes stimulate the activity of the CYPs,
allowing for a reduction in the amount of enzyme required
per reaction (BD GENTEST). The assays monitored, via
fluorescence detection, the formation of a fluorescent
metabolite following incubation of the microsomes with
a specific CYP substrate. Two CYP substrates were tested
for CYP3A4 (7-benzyloxy-4-trifluoromethylcoumarin
and 7-benzyloxyquinoline), as this enzyme has been
shown to exhibit complex inhibition kinetics36 Reactions
(0.2 mL) are performed in 96-well microtiter plates at
37 ꢁC in the presence of an NADPH regenerating system
[b-nicotinamide adenine dinucleotide phosphate
(NADP+), glucose-6-phosphate (G6P), glucose-6-phos-
phate dehydrogenase (G6PDH)] and MgCl2. Inhibition
of metabolic product formation by the test compound
for each enzyme was tested in the absence and presence
of 0.0457–100 lM of test compound. An enzyme-selective
inhibitor was also tested (at eight concentrations) in each
assay as a positive control. All determinations were
performed in duplicate. Assays for all enzymes were per-
formed in the following manner: the NADPH regenerat-
ing system, appropriate buffer solution and vehicle,
inhibitor (positive control) solution or test compound
solution were dispensed into 96-well microtiter plates.
Eight inhibitor and test compound concentrations were
tested using 3-fold serial dilutions. The microtiter plates
containing 0.1 mL/well of the latter mixture was pre-
warmed to 37 ꢁC in an incubator. A solution of buffer,
microsomes, and substrate were separately prepared
and vortex mixed to disperse the protein. The reactions
were initiated by the addition of the microsome/substrate
solution (0.1 mL) to the wells of the microtiter plates con-
taining the pre-warmed NADPH regenerating system,
buffer, and inhibitor ortest compound solutions. The final
concentration of acetonitrile, the vehicle used for the
known inhibitors, was 2%. Following specified incuba-
tion times, the reactions were stopped by the addition of
0.075 mL of a STOP solution (see below). Blank (back-
ground noise) samples were also assayed by adding the
STOP solution prior to the addition of the microsome/
substrate mix to the NADPH regenerating system. The
amount of metabolic product formed was quantified by
fluorescence detection in a fluorescence plate reader utiliz-
ing excitation and emission filters that have been opti-
mized for the detection of each metabolite.
7. Experimental
7.1. Cell lines and cell culture
CEM and CEM/VLB500 human lymphoma cells were
kindly provided by Dr. W. T. Beck (Department of
Biopharmaceutical Sciences, College of Pharmacy, Uni-
versity of Illinois at Chicago). Both cell lines were grown
in RPMI 1640 medium supplemented with 2.0 mM
glutamine and 10% fetal bovine serum (Hyclone, Road
Logan, Utah). Cells were cultured in a moisture-saturat-
ed atmosphere at 37 ꢁC in 5% CO2.
7.2. MDR inhibition assay
Cells were plated in 96-well microtiter plates at 1 · 104
cells/well in media with and without vinblastine or dau-
norubicin (100 nM). Then, anthranilamide modulators
dissolved in DMSO were added at escalating concentra-
tions ranging from 0.025 to 15 lM to the cells. The final
concentration of DMSO in each well was maintained at
maximum 0.5%. The plates were incubated for 3 days.
The cell survival was assessed using the resazurin
assay.24 Briefly, cells were washed three times with
200 lL of PBS solution. Fifty microliters of a PBS solu-
tion containing 20% resazurin in RPMI-1640 were add-
ed. Cell survival was calculated from fluorescence
(excitation, 485 nm; emission, 590 nm) measured with
a FL 600 Reader (Bio-Tek Instruments). Cytotoxicity
was expressed as the concentration of the drug required
to inhibit cell growth by 50% (GI50). Values are the
means of at least three independent determinations.
The values were electronically processed to determine
‘difference scores,’ that is defined as percent survival
of cells treated with test agent alone minus percent sur-
vival of cells treated with test compound and 100 nM
vinblastine. All experiments were performed in quadru-

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P. Labrie et al. / Bioorg. Med. Chem. 14 (2006) 7972–7987
CYP1A2
Microsomes: CYP1A2 (0.5 pmol) and supplemental cDNA-expressed human reductase
NADPH regenerating system: 1.3 mM NADP+, 3.3 mM G6P, 0.4 U/mL G6PDH, 3.3 mM MgCl2
Buffer: 100 mM potassium phosphate buffer, pH 7.4 (PPB)
Inhibitor: furafylline (0.0457–100 lM)
Substrate: 3-cyano-7-ethoxycoumarin (5 lM CEC)
Metabolic product: 3-cyano-7-hydroxycoumarin (CHC)
Incubation time: 15 min
STOP buffer: 80% acetonitrile/20% 0.5 M Tris base
Excitation filter: 400 nm
Emission filter: 460 nm
CYP2A6
Microsomes: CYP2A6 (1.0 pmol) and supplemental cDNA-expressed human reductase and human
cytochrome b5
NADPH regenerating system: 0.065 mM NADP+, 3.3 mM G6P, 0.4 U/mL G6PDH,
3.3 mM MgCl2
Buffer: 100 mM Tris buffer, pH 7.5
Inhibitor: tranylcypromine (0.0457–100 lM)
Substrate: coumarin (3 lM)
Metabolic product: 7-hydroxycoumarin (7-HC, umbelliferone)
Incubation time: 15 min
STOP buffer: 80% acetonitrile/20% 0.5 M Tris base
Excitation filter: 400 nm
Emission filter: 460 nm
CYP2B6
CYP2C8
CYP2C9
Microsomes: CYP2B6 (1.0 pmol) and supplemental cDNA-expressed human reductase and human
cytochrome b5
NADPH regenerating system: 1.3 mM NADP+, 3.3 mM G6P, 0.4 U/mL G6PDH, 3.3 mM MgCl2
Buffer: 100 mM PPB
Inhibitor: tranylcypromine (0.057–125 lM)
Substrate: 7-ethoxy-4-trifluoromethylcoumarin (2.5 lM EFC)
Metabolic product: 7-hydroxy-4-trifluoromethylcoumarin (HFC)
Incubation time: 30 min
STOP buffer: 80% acetonitrile/20% 0.5 M Tris base
Excitation filter: 400 nm
Emission filter: 530 nm
Microsomes: CYP2C8 (4.0 pmol) and supplemental cDNA-expressed human reductase and human
cytochrome b5
NADPH regenerating system: 1.3 mM NADP+, 3.3 mM G6P, 0.4 U/mL G6PDH, 3.3 mM MgCl2
Buffer: 50 mM PPB
Inhibitor: quercetin (0.009–20 lM)
Substrate: dibenzylfluorescein (1 lM DBF)
Metabolic product: fluorescein
Incubation time: 30 min
STOP Buffer: 2 mM NaOH
Excitation filter: 485 nm
Emission filter: 530 nm
Microsomes: CYP2C9*1 (1.0 pmol) and supplemental cDNA-expressed human reductase
and human cytochrome b5 (CYP2C9*1 is the most common allele in human populations which
has been studied to date)
NADPH regenerating system: 1.3 mM NADP+, 3.3 mM G6P, 0.4 U/mL G6PDH, 3.3 mM MgCl2
Buffer: 25 mM PPB
Inhibitor: sulfaphenazole (0.00457–10 lM)
Substrate: 7-methoxy-4-trifluoromethylcoumarin (75 lM MFC)
Metabolic product: 7-hydroxy-4-trifluoromethylcoumarin
Incubation time: 45 min
STOP buffer: 80% acetonitrile/20% 0.5 M Tris base

P. Labrie et al. / Bioorg. Med. Chem. 14 (2006) 7972–7987
7979
Excitation filter: 400 nm
Emission filter: 530 nm
CYP2C19
CYP2D6
CYP2E1
Microsomes: CYP2C19 (0.5 pmol) and supplemental cDNA-expressed human reductase and human
cytochrome b5
NADPH regenerating system: 1.3 mM NADP+, 3.3 mM G6P, 0.4 U/mL G6PDH, 3.3 mM MgCl2
Buffer: 50 mM PPB
Inhibitor: tranylcypromine (0.229–500 lM)
Substrate: 3-cyano-7-ethoxycoumarin (25 lM)
Metabolic product: 3-cyano-7-hydroxycoumarin
Incubation time: 30 min
STOP buffer: 80% acetonitrile/20% 0.5 M Tris base
Excitation filter: 400 nm
Emission filter: 460 nm
Microsomes: CYP2D6*1 (1.5 pmol) and supplemental cDNA-expressed human reductase
(CYP2D6*1 is the most common human CYP2D6 allele)
NADPH regenerating system: 8.2 lM NADP+, 0.41 mM G6P, 0.4 U/mL G6PDH, 0.41 mM MgCl2
Buffer: 100 mM PPB
Inhibitor: quinidine (0.00023–0.5 lM)
Substrate: 3-[2-(N,N-diethyl-N-methylamino)ethyl]-7-methoxy-4-methylcoumarin (1.5 lM AMMC)
Metabolic product: 3-[2-(N,N-diethylamino)ethyl]-7-hydroxy-4-methylcoumarin (AHMC)
Incubation time: 30 min
STOP buffer: 80% acetonitrile/20% 0.5 M Tris base
Excitation filter: 400 nm
Emission filter: 460 nm
Microsomes: CYP2E1 (2.0 pmol) and supplemental cDNA-expressed human reductase
and cytochrome b5
NADPH regenerating system: 1.3 mM NADP+, 3.3 mM G6P, 0.4 U/mL G6PDH, 3.3 mM MgCl2
Buffer: 100 mM PPB
Inhibitor: diethyldithiocarbamic acid (0.0457–100 lM DDTC)
Substrate: 7-methoxy-4-trifluoromethylcoumarin (100 lM MFC)
Metabolic product: HFC
Incubation time: 45 min
STOP buffer: 80% acetonitrile/20% 0.5 M Tris base
Excitation filter: 400 nm
Emission filter: 530 nm
CYP3A4/BFCMicrosomes: CYP3A4 (1.0 pmol) and supplemental cDNA-expressed human reductase
and cytochrome b5
NADPH regenerating system: 1.3 mM NADP+, 3.3 mM G6P, 0.4 U/mL G6PDH, 3.3 mM MgCl2
Buffer: 200 mM PPB
Inhibitor: ketoconazole (0.00229–5 lM)
Substrate: 7-benzyloxy-4-trifluoromethylcoumarin (50 lM BFC)
Metabolic product: 7-hydroxy-4-trifluoromethylcoumarin
Incubation time: 30 min
STOP buffer: 80% acetonitrile/20% 0.5 M Tris base
Excitation filter: 400 nm
Emission filter: 530 nm
CYP3A4/BQ Microsomes: CYP3A4 (3.0 pmol) and supplemental cDNA-expressed human reductase
and cytochrome b5
NADPH regenerating system: 1.3 mM NADP+, 3.3 mM G6P, 0.4 U/mL G6PDH, 3.3 mM MgCl2
Buffer: 200 mM PPB
Inhibitor: ketoconazole (0.00229–5 lM)
Substrate: 7-benzyloxyquinoline (40 lM BQ)
Metabolic product: quinolinol
Incubation time: 30 min

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P. Labrie et al. / Bioorg. Med. Chem. 14 (2006) 7972–7987
STOP buffer: 80% acetonitrile/20% 0.5 M Tris base
Excitation filter: 400 nm
Emission filter: 530 nm
7.4. Chemistry
ArH), 3.85 (s, 6H, OMe), 3.71 (s, 4H, CH2), 2.86 (s,
4H, CH2); 13C NMR (50.3 MHz, CDCl3): 147.8,
147.4, 125.0, 124.9, 114.7, 111.3, 109.3, 55.9, 55.8,
53.8, 49.8, 45.9, 28.5.
Melting points were determined on an electrothermal
melting point apparatus. All nominal and accurate
mass electronic impact (EI) measurements were made
using a JEOL HX110 double focusing mass spectrom-
eter. All nominal and accurate mass electrospray ion-
ization (ESI) measurements were made with a Waters/
Micromass QTOF Ultima Global instrument of
QQTof geometry. IR spectra were recorded on a
7.4.1.3. Preparation of 3-[[2-(3,4-dimethoxyphenyl)-
ethyl](methyl)amino]propanenitrile (8a). Yield: 94%,
yellow oil; IR (NaCl, cmÀ1): 3060, 2939, 2801 (C–H),
2248 (CN); 1590, 1516, 1418 (C@C Ar); 1262 (tertiary
amine); 1H NMR (300 MHz, CDCl3); 6.54 (m, 3H,
ArH), 3.62 (s, 3H, OMe), 3.58 (s, 3H, OMe), 2.47 (m,
6H, CH2), 2.20 (t, 2H, CH2, J = 6.7 Hz), 2.09 (s, 3H,
CH3); 13C NMR (50.3 MHz, CDCl3); 148.8, 147.4,
132.6, 120.5, 119.0, 112.1, 111.3, 59.1, 55.9, 55.8, 52.7,
41.6, 33.4, 16.1.
1
Bomem MB-100 spectrometer. H NMR spectra were
determined at 200 MHz using a Varian XL-200 or at
300 MHz using a Bruker AC-300 spectrometer; 13C
NMR spectra were determined at 50.3 MHz using a
Varian XL-200 or at 75.5 MHz using a Bruker AC-
300 spectrometer; chemical shifts (d) given using
chloroform-d as a reference. The silical gel used for
chromatography is Kieselgel 60 70–230 mesh Merck.
Unless otherwise noted all reagents and solvents ob-
tained from commercial suppliers were used without
further purification.
7.4.1.4. Preparation of 3-(6,7-dimethoxy-3,4-dihydro-
isoquinolin-2(1H)-yl)propanenitrile (8b). Yield: 29%, yel-
low oil; IR (NaCl, cmÀ1); 3055, 2930, 2832 (C–H); 2249
(CN); 1604, 1517, 1464 (C@C Ar); 1264 (tertiary amine);
1H NMR (300 MHz, CDCl3); 6.50 (s, 1H, ArH), 6.43 (s,
1H, ArH), 3.73 (s, 6H, OMe), 3.50 (s, 2H, CH2), 2.66
(m, 6H, CH2), 2.48 (t, 2H, CH2, 6.9 Hz); 13C NMR
(50.3 MHz, CDCl3); 147.6, 147.3, 125.8, 125.7, 118.8,
111.4, 109.4, 55.9, 55.8, 54.9, 53.0, 50.5, 28.4, 16.2.
7.4.1. General procedure for the synthesis of compound
7a–11a and 7b–11b. To a solution of tert-butanol, DMF
or acetonitrile (20–40 mL) containing the amine
(1 equiv) were added sodium iodide (0.5 equiv), sodium
carbonate or potassium carbonate (2 equiv) and the cor-
responding chloronitrile (1.2 equiv). The mixture was
stirred under reflux for 24 h (TLC monitoring, dichloro-
methane/methanol 9:1). The mixture was poured into
20 mL of water and extracted with ethyl acetate. The
organic layers were combined and evaporated. The
crude product was dissolved in 1 N HCl (10–20 mL)
and extracted with ethyl acetate. The aqueous layers
were combined and NaOH pellets were added to alkalin-
ize the mixture. The white solution was extracted with
ethyl acetate. The organic layer was washed with brine,
dried with anhydrous sodium sulfate, filtered, and evap-
orated under vacuum. Pure product was obtained after
column chromatography on silica gel using a 0%–5%
methanol/dichloromethane as elution solvent.
7.4.1.5. Preparation of 4-[[2-(3,4-dimethoxyphenyl)-
ethyl](methyl)amino]butanenitrile (9a). Yield: 45%, yel-
low oil; IR (NaCl, cmÀ1); 2924, 2850 (C–H); 1591,
1
1516, 1460 (C@C Ar); 1264 (tertiary amine); H NMR
(200 MHz, CDCl3); 6.70 (m, 3H, ArH), 3.83 (s, 3H,
OMe), 3.80 (s, 3H, OMe), 2.65 (m, 2H, CH2), 2.55 (m,
2H, CH2), 2.48 (m, 2H, CH2), 2.29 (m, 2H, CH2), 2.23
(s, 3H, CH3), 1.72 (m, 2H, CH2); 13C NMR
(50.3 MHz, CDCl3); 148.8, 147.3, 132.9, 120.5, 119.8,
112.1, 111.3, 59.5, 55.9, 55.8, 55.4, 41.9, 33.4, 23.4, 14.6.
7.4.1.6. Preparation of 4-(6,7-dimethoxy-3,4-dihydro-
isoquinolin-2(1H)-yl) butanenitrile (9b). Yield: 65%, yel-
low solid; IR (NaCl, cmÀ1); 3046, 2929, 2846 (C–H);
1
1612, 1517, 1465 (C@C Ar); 1265 (tertiary amine); H
NMR (200 MHz, CDCl3); 6.58 (s, 1H, ArH), 6.50 (s,
1H, ArH), 3.82 (s, 3H, OMe), 3.81 (s, 3H, OMe), 3.52
(s, 2H, CH2), 2.82 (m, 2H, CH2), 2.68 (m, 4H, CH2),
2.45 (t, 2H, CH2, J = 6.9 Hz), 1.89 (m, 2H, CH2); 13C
NMR (50.3 MHz, CDCl3); 147.6, 147.3, 126.3, 126.1,
119.8, 111.4, 109.4, 55.9, 55.6, 50.9, 28.6, 23.1, 14.8.
7.4.1.1. Preparation of [[2-(3,4-dimethoxyphenyl)eth-
yl](methyl)amino]acetonitrile (7a). Yield: 84%, yellow oil;
IR (NaCl, cmÀ1); 2948, 2835, 2803 (C–H); 2229 (CN);
1591, 1516, 1464, 1418 (C@C Ar); 1264 (tertiary amine);
1H NMR (300 MHz, CDCl3); 6.70 (m, 3H, ArH), 3.74
(s, 3H, OMe), 3.70 (s, 3H, OMe), 3.41 (s, 2H, CH2),
2.58 (s, 4H, CH2), 2.26 (s, 3H, CH3); 13C NMR
(75.5 MHz, CDCl3); 148.9, 147.5, 131.9, 120.5, 114.8,
111.9, 111.4, 57.4, 55.8, 55.7, 45.1, 41.9, 33.5.
7.4.1.7. Preparation of 5-[[2-(3,4-dimethoxyphenyl)eth-
yl](methyl)amino]pentanenitrile (10a). Yield: 89%, sticky
yellow oil. IR (NaCl, cmÀ1); 3046, 2954, 2838, 2797
(C–H), 2247 (CN); 1591, 1516, 1465, 1421 (C@C Ar);
1262 (tertiary amine); 1H NMR (400 MHz, CDCl3);
6.75 (m, 3H, ArH), 3.87 (s, 3H, OMe), 3.85 (s, 3H,
OMe), 2.70 (m, 2H, CH2), 2.57 (m, 2H, CH2), 2.41 (m,
2H, CH2), 2.33 (m, 2H, CH2), 2.28 (s, 3H, CH3), 1.63
(m, 4H, CH2); 13C NMR (50.3 MHz, CDCl3); 148.8,
7.4.1.2. Preparation of (6,7-dimethoxy-3,4-dihydroiso-
quinolin-2(1H)-yl)acetonitrile (7b). Yield: 76%, yellow
oil; IR (NaCl, cmÀ1): 3051, 2929, 2850 (C–H); 1589,
1
1515, 1464 (C@C Ar); 1264 (amine tertiaire); H NMR
(200 MHz, CDCl3): 6.60 (s, 1H, ArH), 6.53 (s, 1H,

P. Labrie et al. / Bioorg. Med. Chem. 14 (2006) 7972–7987
7981
147.3, 133.0, 120.5, 119.7, 112.0, 111.3, 59.7, 56.4, 55.9,
55.8, 41.9, 33.4, 26.1, 23.3, 16.9.
2927, 2836 (C–H); 1659 (NH2 deformation); 1517,
1464, 1420 (C@C Ar); 1264 (tertiary amine); H NMR
1
(300 MHz, CDCl3) d: 6.71 (m, 3H, ArH), 3.86 (s, 3H,
OMe), 3.83 (s, 3H, OMe), 3.37 (s, 2H, NH2), 2.80 (t,
2H, CH2, J = 5.9 Hz), 2.73 (m, 2H, CH2), 2.70 (m, 2H,
CH2), 2.52 (m, 2H, CH2), 2.29 (s, 3H, CH3); 13C
NMR (75.5 MHz, CDCl3) d: 148.9, 147.3, 132.9,
120.6, 120.1, 112.1, 111.3, 59.7, 58.3, 55.9, 41.9, 38.7,
33.4.
7.4.1.8. Preparation of 5-(6,7-dimethoxy-3,4-dihydro-
isoquinolin-2(1H)-yl)pentanenitrile (10b). Yield: 46%, as
sticky yellow oil; IR (NaCl, cmÀ1); 3056, 2925, 2851
(C–H); 1611, 1517, 1465 (C@C Ar); 1265 (tertiary
amine); 1H NMR (300 MHz, CDCl3); 6.58 (s, 1H,
ArH), 6.50 (s, 1H, ArH), 3.82 (s, 3H, OMe), 3.81 (s,
3H, OMe), 3.54 (s, 2H, CH2), 2.79 (m, 2H, CH2), 2.68
(m, 2H, CH2), 2.52 (m, 2H, CH2), 2.39 (m, 2H, CH2),
1.72 (m, 4H, CH2); 13C NMR (50.3 MHz, CDCl3);
147.5, 147.1, 126.5, 126.1, 119.7, 111.4, 109.4, 56.9,
55.9, 55.8, 55.7, 50.9, 28.6, 25.9, 23.3, 16.9.
7.4.2.2. Preparation of 2-(6,7-dimethoxy-3,4-dihydro-
isoquinolin-2(1H)-yl)ethanamine (12b). To a 50 mL solu-
tion of 1.4 M NaOH/EtOH containing 7b (1.19 mmol)
was added Raney nickel (150 mg, wet) to give 0.243 g
of compound 12b as yellow oil after hydrogenation
(86%). IR (NaCl, cmÀ1); 3367 (NH2); 3053, 2926
(C–H); 1612 (NH2 deformation) 1517, 1465, (C@C
Ar); 1264 (tertiary amine); 738 (CH deformation Ar);
1H NMR (300 MHz, CDCl3) d: 6.47 (s, 1H, ArH),
6.40 (s, 1H, ArH), 3.71 (s, 3H, OMe), 3.70 (s, 3H,
OMe), 3.43 (s, 2H, CH2), 2.73 (m, 4H, CH2), 2.59 (m,
2H, CH2), 2.43 (m, 2H, CH2), 1.56 (s, 2H, NH2); 13C
NMR (75.5 MHz, CDCl3) d: 147.5, 147.2, 126.6,
126.2, 111.4, 109.5, 60.8, 55.9, 55.8, 51.0, 39.1, 29.6,
28.6.
7.4.1.9. Preparation of 6-[[2-(3,4-dimethoxyphenyl)-
ethyl](methyl)amino]hexanenitrile, (11a). Yield: 46%, yel-
low oil; IR (NaCl, cmÀ1); 2938, 2861, (C–H); 2243 (CN);
1592, 1517, 1465, 1422 (C@C Ar); 1264 (tertiary amine);
1H NMR (300 MHz, CDCl3); 6.69 (m, 3H, ArH), 3.82
(s, 3H, OMe), 3.79 (s, 3H, OMe), 2.67 (m, 2H, CH2),
2.55 (m, 2H, CH2), 2.37 (t, 2H, CH2, J = 6.7 Hz), 2.30
(m, 5H, CH2 and CH3), 1,61 (m, 2H, CH2), 1.43 (m,
4H, CH2); 13C NMR (75.5 MHz, CDCl3); 148.8,
147.3, 132.9, 120.5, 119.7, 112.1, 111.3, 59.7, 57.1,
55.9, 55.8, 42.0, 33.3, 26.5, 26.4, 25.3, 17.0.
7.4.2.3. Preparation of N-[2-(3,4-dimethoxyphenyl)eth-
yl]-N-methylpropane-1,3-diamine (13a). To a 75 mL
solution of 1.4 M NaOH/EtOH containing 8a
(1.95 mmol) was added 150 mg of Raney nickel wet to
give 0.489 g of compound 13a as yellow oil after hydro-
genation (99%). IR (NaCl, cmÀ1) d: 3425 (NH2); 2928,
2851 (C–H); 1649 (NH2 deformation); 1517, 1464,
1422 (C@C Ar); 1264 (tertiary amine); 1H NMR
(300 MHz, CDCl3) d: 6.46 (m, 3H, ArH), 3.57 (s, 3H,
OMe), 3.54 (s, 3H, OMe), 2.44 (m, 4H, CH2), 2.32 (m,
2H, CH2), 2.17 (t, 2H, CH2, J = 6.9 Hz), 2.01 (s, 3H,
CH3), 1.95 (s, 2H, NH2), 1.35 (m, 2H, CH2); 13C
NMR (75.5 MHz, CDCl3) d: 148.6, 147.1, 132.9,
120.3, 111.9, 111.2, 59.7, 55.6, 55.5, 55.3, 41.9, 40.4,
33.2, 30.6.
7.4.1.10. Preparation of 6-(6,7-dimethoxy-3,4-dihydro-
isoquinolin-2(1H)-yl)hexanenitrile (11b). Yield: 38%, yel-
low oil; IR (NaCl, cmÀ1); 2921, 2861 (C–H); 1517, 1464,
(C@C Ar); 1265 (tertiary amine); 737 (CH deformation
1
Ar); H NMR (300 MHz, CDCl3); 6.50 (s, 1H, ArH),
6.44 (s, 1H, ArH), 3.72 (s, 6H, OMe), 3.48 (s, 2H,
CH2), 2.73 (t, 2H, CH2, J = 4.9 Hz), 2.63 (t, 2H, CH2,
J = 5.7 Hz), 2.43 (t, 2H, CH2, J = 7.1 Hz), 2.25 (t, 2H,
CH2, J = 7.1 Hz), 1.57 (m, 4H, CH2), 1.41 (m, 2H,
CH2); 13C NMR (75.5 MHz, CDCl3); 147.6, 147.2,
126.1, 125.9, 119.7, 111.4, 109.5, 57.6, 55.9, 55.8, 55.5,
50.9, 28.4, 26.5, 26.2, 25.2, 17.0.
7.4.2. General procedure for the synthesis of compounds
12a–16a and 12b–16b. Compounds (7a–11a and 7b–11b)
(1 equiv) were dissolved in 20–100 mL of a solution of
freshly prepared 1.4 M NaOH in ethanol. To this solu-
tion was added wet Raney nickel (150–200 mg). The
bottle was placed in a Parr shaker apparatus and the
system was purged three times with hydrogen. The sys-
tem pressure was adjusted to 40 psi and agitated for
12 h. The reaction mixture was filtered through CeliteTM
to remove the Raney nickel. The CeliteTM pad was
washed three times with ethanol and the filtrate was
evaporated to a small volume and diluted with 40 mL
of distilled water, alkalized with NaOH pellets and
extracted with chloroform (3· 30 mL). The organic
phases were combined, dried over anhydrous Na2SO4,
filtered and evaporated under vacuum.
7.4.2.4. Preparation of 3-(6,7-dimethoxy-3,4-dihydro-
isoquinolin-2(1H)-yl)propan-1-amine (13b). To a 50 mL
solution of 1.4 M NaOH/EtOH containing 8b
(1.04 mmol) was added 150 mg of Raney nickel wet to
give 0.227 g of compound 13b as yellow oil after hydro-
genation (87%). IR (NaCl, cmÀ1); 3396 (NH2); 2928,
2856 (C–H); 1638 (NH2 deformation); 1517, 1463,
(C@C Ar); 1264 (tertiary amine); 737 (CH deformation
1
Ar); H NMR (300 MHz, CDCl3) d: 6.56 (s, 1H, ArH),
6.49 (s, 1H, ArH), 3.80 (s, 6H, OMe), 3.52 (s, 2H, CH2),
2.77 (m, 4H, CH2), 2.68 (t, 2H, CH2, J = 5.5 Hz), 2.54 (t,
2H, CH2, J = 7.2 Hz), 1.72 (m, 2H, CH2); 13C NMR
(75.5 MHz, CDCl3) d: 147.5, 147.2, 126.7, 126.3, 111.4,
109.5, 56.2, 55.9, 51.0, 40.7, 30.7, 29.6, 28.7.
7.4.2.1. Preparation of N-[2-(3,4-dimethoxyphenyl)eth-
yl]-N-methylethane-1,2-diamine (12a). To a 100 mL solu-
tion of 1.4 M NaOH in ethanol containing 7a
(6.447 mmol) was added Raney nickel wet (500 mg) to
give compound 12a as yellow oil after hydrogenation
(1.455 g, 95%). IR (NaCl, cmÀ1): 3381 (NH2); 3052,
7.4.2.5. Preparation of N-[2-(3,4-dimethoxyphenyl)eth-
yl]-N-methylbutane-1,4-diamine (14a). To a 50 mL solu-
tion of 1.4 M NaOH in ethanol containing 9a (1.10
mmol) was added Raney nickel (150 mg, wet) to give
0.268 g of compound 14a as yellow oil after hydrogena-
tion (91%). IR (NaCl, cmÀ1): 3357 (NH2); 3055, 2932,

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P. Labrie et al. / Bioorg. Med. Chem. 14 (2006) 7972–7987
2852, 2798 (C–H); 1590 (NH2 deformation); 1515, 1465,
1418 (C@C Ar); 1265 (tertiary amine); 737 (CH defor-
(C–H); 1615 (NH2 deformation), 1517, 1463, (C@C
Ar); 1264 (tertiary amine); 739 (CH deformation Ar);
1H NMR (300 MHz, CDCl3) d: 6.73 (m, 3H, ArH),
3.84 (s, 3H, OMe), 3.82 (s, 3H, OMe), 2.62 (m, 6H,
CH2), 2.36 (m, 2H, CH2), 2.27 (s, 3H, CH3), 1.32 (m,
10H, NH2 + CH2); 13C NMR (75.5 MHz, CDCl3) d:
148.8, 147.3, 132.9, 120.5, 119.7, 112.1, 111.3, 59.7,
57.1, 55.9, 55.8, 42.0, 33.3, 26.5, 26.4, 25.3, 17.0.
1
mation Ar); H NMR (200 MHz, CDCl3) d: 6.65 (m,
3H, ArH), 3.75 (s, 3H, OMe), 3.72 (s, 3H, OMe), 2.57
(m, 6H, CH2), 2.29 (m, 2H, CH2), 2.18 (s, 3H, CH3),
1.96 (bs, 2H, NH2), 1.36 (m, 4H, CH2); 13C NMR
(75.5 MHz, CDCl3) d: 148.7, 147.2, 133.0, 120.5, 112.0,
111.2, 59.7, 57.4, 55.8, 55.7, 42.1, 41.9, 33.3, 31.5, 24.6.
7.4.2.6. Preparation of 4-(6,7-dimethoxy-3,4-dihydro-
isoquinolin-2(1H)-yl)butan-1-amine (14b). To a 50 mL
solution of 1.4 M NaOH in ethanol containing 9b
(0.865 mmol) was added Raney nickel (150 mg, wet) to
give 0.229 g of compound 14b as yellow oil after hydro-
genation (99%). IR (NaCl, cmÀ1); 3430 (NH2); 2924,
2853 (C–H); 1630 (NH2 deformation); 1518, 1463,
(C@C Ar); 1264 (tertiary amine); 739 (CH deformation
7.4.2.10. Preparation of 6-(6,7-dimethoxy-3,4-dihydro-
isoquinolin-2(1H)-yl)hexan-1-amine (16b). To a 50 mL
solution of 1.4 M NaOH in ethanol containing 11b
(0.869 mmol) was added Raney nickel (150 mg, wet) to
give 0.219 g of compound 16b as yellow oil after hydro-
genation (86%). IR (NaCl, cmÀ1); 3445 (NH2); 2929,
2854 (C–H); 1612 (NH2 deformation), 1517, 1465,
(C@C Ar); 1264 (tertiary amine); 740 (CH deformation
1
1
Ar); H NMR (300 MHz, CDCl3) d: 6.42 (s, 1H, ArH),
Ar); H NMR (300 MHz, CDCl3) d: 6.38 (s, 1H, ArH),
6.35 (s, 1H, ArH), 3.66 (s, 6H, OMe), 3.04 (br s, 4H,
CH2 + NH2), 2.66 (t, 2H, CH2, J = 5.5 Hz), 2.56 (m,
4H, CH2), 2.33 (t, 2H, CH2, 7.3 Hz), 1.40 (m, 4H,
CH2); 13C NMR (50.3 MHz, CDCl3) d: 147.4, 147.0,
126.2, 126.0, 111.2, 109.4, 57.9, 57.1, 55.7, 55.5, 50.8,
41.6, 31.1, 28.2, 24.2.
6.32 (s, 1H, ArH), 3.61 (s, 6H, OMe), 3.33 (s, 2H, CH2),
2.60 (t, 2H, CH2, J = 5.2 Hz), 2.47 (t, 2H, CH2,
J = 5.6 Hz), 2.27 (t, 2H, CH2, J = 7.5 Hz), 1.83 (s, 2H,
NH2), 1.40 (m, 2H, CH2), 1.20 (m, 6H, CH2); 13C
NMR (75.5 MHz, CDCl3) d: 147.4, 147.1, 126.7,
126.1, 111.3, 109.5, 58.2, 55.7, 50.9, 41.8, 33.3, 28.6,
27.3, 27.1, 26.7.
7.4.2.7. Preparation of N-[2-(3,4-dimethoxyphenyl)eth-
yl]-N-methylpentane-1,5-diamine (15a). To a 100 mL
solution of 1.4 M NaOH in ethanol containing 10a
(3.07 mmol) was added Raney nickel (300 mg, wet) to
give 0.689 g of compound 15a as yellow oil after hydro-
genation (80%). IR (NaCl, cmÀ1); 3347 (NH2); 2930,
2856 (C–H); 1649 (NH2 deformation), 1515, 1465,
(C@C Ar); 1264 (tertiary amine); 733 (CH deformation
Ar); 1H NMR (300 MHz, CDCl3) d: 6.56 (m, 3H, ArH),
3.68 (s, 3H, OMe), 3.64 (s, 3H, OMe), 2.49 (m, 4H,
CH2), 2.39 (m, 2H, CH2), 2.21 (t, 2H, CH2,
J = 7.3 Hz), 2.10 (s, 3H, CH3), 1.75 (s, 2H, NH2), 1.30
(m, 4H, CH2), 1.20 (m, 2H, CH2); 13C NMR
(75.5 MHz, CDCl3) d: 148.7, 147.2, 133.1, 120.4, 112.0,
111.2, 59.7, 57.5, 55.7, 55.6, 42.0, 41.9, 33.4, 27.1, 24.7.
7.4.3. General procedure for the synthesis of compounds
17a–21a and 17b–21b. To a solution of acetonitrile
(15 mL) containing the diamine (8a–16a or 8b–16b)
(1 equiv) was added isantoic anhydride (1 equiv) and
the mixture was stirred under reflux for 18 h. The mix-
ture was diluted with 20 mL AcOEt and extracted with
HCl 1N. The aqueous layer was combined and alkalized
with NaOH pellets. The white solution was extracted
with AcOEt. The organic phases were combined, dried
over anhydrous Na2SO4, filtered, and concentrated un-
der vacuum. Pure product was obtained after column
chromatography on silica gel using a 0% to 5% metha-
nol/dichloromethane as elution solvent.
7.4.3.1. Preparation of 2-amino-N-{2-[[2-(3,4-dimeth-
oxyphenyl)ethyl](methyl)amino]ethyl} benzamide (17a).
7.4.2.8. Preparation of 5-(6,7-dimethoxy-3,4-dihydro-
isoquinolin-2(1H)-yl)pentan-1-amine (15b). To a 50 mL
solution of 1.4 M NaOH in ethanol containing 10b
(1.02 mmol) was added Raney nickel (150 mg, wet) to
give 0.248 g of compound 15b as yellow oil after hydro-
genation (87%). IR (NaCl, cmÀ1); 3391 (NH2); 3053,
2930, 2855 (C–H); 1640 (NH2 deformation), 1517,
1465, (C@C Ar); 1265 (tertiary amine); 737 (CH defor-
mation Ar); 1H NMR (300 MHz, CDCl3) d: 6.53 (s,
1H, ArH), 6.47 (s, 1H, ArH), 3.78 (s, 6H, OMe), 3.49
(s, 2H, CH2), 2.76 (m, 2H, CH2), 2.65 (m, 4H, CH2),
2.44 (m, 2H, CH2), 2.20 (2H, NH2), 1.38 (m, 6H,
CH2); 13C NMR (75.5 MHz, CDCl3) d: 147.4, 147.1,
126.7, 126.2, 111.4, 109.5, 58.3, 55.8, 55.7, 51.0, 41.8,
33.1, 28.6, 27.0, 24.8.
Yield: 84%, yellow oil; IR (NaCl, cmÀ1
) 3362
(NHCO + NH2); 2921, 2851 (C–H); 1642 (CO); 1588
(NH2 deformation); 1515, 1465, (C@C Ar); 1264 (tertia-
ry amine); 735 (CH deformation Ar); 1H NMR
(300 MHz, CDCl3) d: 7.10 (t, 1H, ArH, J = 6.9 Hz and
J = 14.6 Hz), 7.04 (d, 1H, ArH, J = 6.9 Hz), 6.66 (m,
4H, ArH + 1H CONH), 6.55 (m, 2H, ArH), 5.55 (s,
2H, NH2), 3.77 (s, 3H, OMe), 3.73 (s, 3H, OMe), 3.40
(m, 2H, CH2), 2.66 (m, 6H, CH2), 2.26 (s, 3H, CH3);
13C NMR (75.5 MHz, CDCl3) d: 169.3, 148.9, 148.8,
147.4, 132.8, 132.1, 127.3, 120.5, 117.1, 116.5, 115.9,
112.0, 111.3, 58.9, 55.9, 55.7 55.5, 41.6, 36.6, 33.2.
7.4.3.2. Preparation of 2-amino-N-[2-(6,7-methoxy-
3,4-dihydroisoquinolin-2(1H)-yl)ethyl] benzamide (17b).
7.4.2.9. Preparation of N-[2-(3,4-dimethoxyphenyl)eth-
yl]-N-methylhexane-1,6-diamine (16a). To a 50 mL solu-
tion of 1.4 M NaOH in ethanol containing 11a (0.262
mmol) was added Raney nickel (150 mg, wet) to give
0.0753 g of compound 16a as yellow oil after hydrogena-
tion (97%). IR (NaCl, cmÀ1); 3445 (NH2); 2923, 2853
Yield 74%, yellow oi;. IR (NaCl, cmÀ1
) 3435
(NHCO + NH2); 2918, 2849 (C–H); 1711 (CO); 1625
(NH2 deformation); 1517, 1445, (C@C Ar); 1222 (tertia-
1
ry amine); H NMR (300 MHz, CDCl3) d: 7.21 (m, 1H,
ArH), 7.08 (m, 1H, ArH), 6.82 (m, 1H, NHCO), 6.50
(m, 4H, ArH), 5.28 (s, 2H, NH2), 3.74 (s, 3H, OMe),

P. Labrie et al. / Bioorg. Med. Chem. 14 (2006) 7972–7987
7983
3.73 (s, 3H, OMe), 3.50 (m, 4H, CH2), 2.68 (m, 6H,
CH2); 13C NMR (75.5 MHz, CDCl3) d: 169.4, 148.7,
147.7, 147.4, 132.1, 127.5, 126.0, 117.2, 116.5, 116.1,
111.5, 109.5, 56.2, 56.0, 55.9, 55.3, 50.6, 36.4, 28.5.
filtered, and concentrated under reduced pressure. The
crude was purified by flash chromatography on silica
gel using a gradient concentration of solvent from 2%
to 10% methanol (2 M NH3) in dichloromethane to give
19 as a yellow oil (0.105 g, 32%). IR (NaCl, cmÀ1): 3057,
2927, 2855 (C–H); 1661 (CO); 1518, 1465, (C@C Ar);
1531, 1349 (NO2); 1265 (tertiary amine); 740 (CH defor-
mation Ar); 1H NMR (300 MHz, CDCl3) d: 8.87 (s, 1H,
NHCO), 7.75 (d, 1H, ArH, J = 6.9 Hz), 7.29 (m, 3H,
ArH), 6.30 (s, 1H, ArH), 6.19 (s, 1H, ArH), 3.79 (s,
3H, OMe), 3.77 (s, 3H, OMe), 3.42 (m, 2H, CH2),
3.35 (s, 2H, CH2), 2.54 (m, 6H, CH2), 1.75 (m, 4H,
CH2); 13C NMR (75.5 MHz, CDCl3) d: 166.6, 147.4,
147.1, 133.6, 133.2, 129.5, 128.3, 125.5, 124.1, 111.0,
109.1, 57.8, 55.9, 55.7, 55.6, 50.1, 40.1, 28.0, 27.8, 25.1.
7.4.3.3. Preparation of 2-amino-N-{2-[[2-(3,4-dimeth-
oxyphenyl)ethyl](methyl)amino]propyl} benzamide (18a).
) 3352
Yield: 85%, yellow oil; IR (NaCl, cmÀ1
(NHCO + NH2); 3055, 2927, 2853 (C–H); 1635 (CO);
1588 (NH2 deformation); 1517, 1465, (C@C Ar); 1264
(tertiary amine); 737 (CH deformation Ar); H NMR
1
(300 MHz, CDCl3) d: 8.05 (m, 1H, NHCO), 7.14 (m,
2H, ArH), 6.67 (m, 4H, ArH), 6.56 (t, 1H, ArH,
J = 7.6 Hz), 5.64 (br s, 2H, NH2), 3.82 (s, 3H, OMe),
3.78 (s, 3H, OMe), 3.47 (q, 2H, CH2, J = 5.6 Hz and
J = 11.3 Hz), 2.64 (m, 6H, CH2), 2.29 (s, 3H, CH3),
1.75 (m, 1H, CH2); 13C NMR (75.5 MHz, CDCl3) d:
169.2, 149.0, 148.9, 147.4, 132.6, 131.9, 127.2, 120.5,
117.2, 116.3, 116.1, 111.9, 111.4, 60.1, 57.3, 55.9, 55.8,
53.5, 41.9, 40.1, 33.4, 25.3.
7.4.3.7. Preparation of 2-amino-N-[2-(6,7-methoxy-
3,4-dihydroisoquinolin-2(1H)-yl)butyl] benzamide (19b).
To a solution of methanol (20 mL) containing 22
(0.0805 g, 0.195 mmol) were added ammonium chloride
(0.0625 g, 1.16 mmol) and dust zinc (0.255 g, 3.89 mol).
The mixture was stirred at refluxed for 90 min. The reac-
tion mixture was filtered and the solvent evaporated un-
der vacuum. The crude was diluted with 1 M HCl
(15 mL) and extracted with ethyl acetate (3· 15 mL).
The aqueous layers were combined, alkalized with
NaOH pellets, and extracted with ethyl acetate (3·
15 mL). The organic layers were combined, dried over
anhydrous Na2SO4, filtered, and evaporated to give
19 b as yellow oil (0.054 g, 72%). IR (NaCl, cmÀ1):
3469 (NHCO); 3352 (NH2); 2928, 2851 (C–H); 1638
(CO); 1584 (NH2 deformation); 1518, 1464, (C@C Ar);
1264 (tertiary amine); 740 (CH deformation Ar); 1H
NMR (300 MHz, CDCl3) d: 7.12 (m, 3H, ArH + NH-
CO), 6.58 (d, 1H, ArH, J = 8.1 Hz), 6.55 (s, 1H, ArH),
6.45 (s, 1H, ArH), 6.30 (t, 1H, ArH, J = 7.5 Hz), 5.50
(s, 2H, NH2), 3.82 (s, 3H, OMe), 3.80 (s, 3H, OMe),
3.50 (s, 2H, CH2), 3.39 (m, 2H, CH2), 2.78 (t, 2H,
CH2, J = 5.7 Hz), 2.68 (t, 2H, CH2, J = 5.5 Hz), 2.52
(t, 2H, CH2, J = 6.5 Hz), 1.70 (m, 4H, CH2); 13C
NMR (75.5 MHz, CDCl3) d: 169.4, 148.6, 147.6,
147.3, 131.9, 127.3, 126.3, 126.1, 117.1, 116.4, 116.3,
111.4, 109.6, 57.3, 55.9, 50.5, 39.5, 28.4, 27.2, 24.8.
7.4.3.4. Preparation of 2-amino-N-[2-(6,7-methoxy-
3,4-dihydroisoquinolin-2(1H)-yl)propyl] benzamide (18b).
Yield: 56%, yellow oil; IR (NaCl, cmÀ1) 3454 (NHCO);
3352 (NH2); 2924, 2841 (C–H); 1636 (CO); 1584 (NH2
deformation); 1518, 1464, (C@C Ar); 1264 (tertiary
amine); 738 (CH deformation Ar); 1H NMR
(300 MHz, CDCl3) d: 8.59 (s, 1H, NHCO), 6.98 (m,
2H, ArH), 6.59 (s, 1H, ArH), 6.52 (d, 1H, ArH,
J = 8.1 Hz), 6.50 (s, 1H, ArH), 5.97 (t, 1H, ArH,
J = 7.8 Hz), 5.71 (s, 2H, NH2), 3.84 (s, 3H, OMe), 3.80
(s, 3H, OMe), 3.66 (s, 2H, CH2), 3.54 (q, 2H, CH2,
J = 5.5 and 10.5 Hz), 2.75 (m, 6H, CH2), 1.85 (m, 2H,
CH2); 13C NMR (75.5 MHz, CDCl3) d: 169.3, 148.9,
147.8, 147.4, 131.8, 127.4, 125.8, 125.7, 117.1, 116.0,
115.5, 111.4, 109.5, 57.9, 56.0, 55.9, 55.5, 51.2, 40.4,
28.5, 24.4.
7.4.3.5. Preparation of 2-amino-N-{2-[[2-(3,4-dimeth-
oxyphenyl)ethyl](methyl)amino]butyl} benzamide (19a).
Yield: 80%, yellow oil; IR (NaCl, cmÀ1) 3454 (NHCO);
3367 (NH2); 2928, 2853 (C–H); 1640 (CO); 1586 (NH2
deformation); 1516, 1464, (C@C Ar); 1264 (tertiary
amine); 738 (CH deformation Ar); 1H NMR
(300 MHz, CDCl3) d: 7.27 (d, 1H, ArH, J = 6.9 Hz),
7.14 (m, 2H, ArH + NHCO), 6.64 (m, 5H, ArH), 5.49,
(s, 2H, NH2), 3.83 (s, 3H, OMe), 3.80 (s, 3H, OMe),
3.36 (m, 2H, CH2), 2.65 (m, 2H, CH2), 2.55 (m, 2H,
CH2), 2.41 (t, 2H, CH2, J = 6.4 Hz), 2.24 (s, 3H, CH3),
1.59 (m, 4H, CH2); 13C NMR (75.5 MHz, CDCl3) d:
169.5, 148.9, 148.7, 147.4, 133.0, 132.0, 127.3, 120.5,
117.2, 116.6, 116.4, 112.1, 111.4, 59.6, 57.1, 55.9, 55.8,
42.2, 39.6, 33.1, 27.5, 25.1.
7.4.3.8. Preparation of 2-amino-N-{2-[[2-(3,4-meth-
oxyphenyl)ethyl](methyl)amino]pentyl} benzamide (20a).
Yield: 82%, yellow oil. IR (NaCl, cmÀ1); 3454 (NHCO);
3347 (NH2); 2926, 2851 (C–H); 1640 (CO); 1516, 1448,
(C@C Ar); 1264 (tertiary amine); 738 (CH deformation
Ar); 1H NMR (300 MHz, CDCl3) d: 7.20 (m, 1H, ArH),
7.10 (t, 1H, ArH, J = 6.8 Hz), 6.60 (m, 5H, ArH), 6.16
(m, 1H, NHCO), 5.42, (s, 2H, NH2), 3.78 (s, 3H,
OMe), 3.76 (s, 3H, OMe), 3.32 (m, 2H, CH2NHCO),
2.65 (m, 2H, CH2), 2.50 (m, 2H, CH2), 2.34 (t, 2H,
CH2, 7.2 Hz), 2.23 (s, 3H, CH3), 1.53 (m, 4H, CH2),
1.33 (m, 2H, CH2); 13C NMR (75.5 MHz, CDCl3) d:
169.4, 148.9, 148.7, 147.4, 133.1, 132.1, 127.1, 120.6,
117.3, 116.5, 116.4, 112.1, 111.3, 59.7, 57.4, 55.9, 55.8,
42.1, 39.6, 33.3, 29.6, 26.8, 24.8.
7.4.3.6. Preparation of 2-nitro-N-[2-(6,7-dimethoxy-
3,4-dihydroisoquinolin-2(1H)-yl)butyl] benzamide (19).
To a solution of ethyl acetate (30 mL) containing com-
pound 14b (0.210 g, 0.796 mmol), were added 2-nitro-
benzoylchloride (110 lL, 0.835 mmol) and DOWEX
1X8 (0.261 g, 100–200 mesh) chloride resin. The mixture
was stirred for overnight at room temperature, filtered,
and extracted with 1 N NaOH (3· 10 mL). The organic
phases were combined, dried over anhydrous Na2SO4,
7.4.3.9. Preparation of 2-amino-N-[2-(6,7-methoxy-
3,4-dihydroisoquinolin-2(1H)-yl)pentyl] benzamide (20b).
Yield: 74%, yellow oil. IR (NaCl, cmÀ1): 3450 (NHCO);

7984
P. Labrie et al. / Bioorg. Med. Chem. 14 (2006) 7972–7987
3347 (NH2); 2929, 2855 (C–H); 1644 (CO); 1585 (NH2
deformation); 1516, 1464, (C@C Ar); 1264 (tertiary
amine); 739 (CH deformation Ar); 1H NMR
(300 MHz, CDCl3) d: 7.25 (m, 1H, ArH), 7.14 (m, 1H,
ArH), 6.55 (m, 3H, ArH), 6.49 (s, 1H, ArH), 6.31 (s,
1H, NHCO), 5.46 (s, 2H, NH2), 3.80 (s, 6H, OMe),
3.52 (s, 2H, CH2), 3.35 (q, 2H, CH2, J = 6.7 Hz and
12.9 Hz), 2.78 (t, 2H, CH2, J = 5.5 Hz), 2.67 (t, 2H,
CH2, J = 5.3 Hz), 2.47 (t, 2H, CH2, J = 7.2 Hz), 1.60
(m, 4H, CH2), 1.40 (m, 2H, CH2); 13C NMR
(75.5 MHz, CDCl3) d: 169.4, 148.6, 147.5, 147.2, 132.1,
127.2, 126.5, 126.1, 117.2, 116.5, 116.4, 111.4, 109.5,
58.1, 55.9, 55.8, 55.7, 51.0, 39.5, 29.6, 28.5, 26.8, 24.8.
7.4.4.1. N-[2-({[{[2-(3,4- methoxyphenyl)ethyl](meth-
yl)amino}ethyl]amino}carbonyl) phenyl]quinoline-3-car-
boxamide (22a). To a solution of dichloromethane
(10 mL) containing 17a (0.119 mmol) were added
3-quinoline acid chloride and DOWEX 1X8 (0.112 g,
100–200 mesh) to give a yellow solid after purification
(0.0425 g, 70%). Mp 149–150 ꢁC; IR (NaCl, cmÀ1):
2925, 2851 (C–H); 1672, 1600 (CO); 1515, 1463, 1448
(C@C Ar); 1264 (tertiary amine); 738 (CH deformation
Ar); 1H NMR (300 MHz, CDCl3) d: 12.79 (s, 1H,
NHCO), 9.55 (d, 1H, ArH, J = 2.08 Hz), 8.84 (d, 1H,
ArH, J = 8.4 Hz), 8.78 (d, 1H, ArH, J = 1.6 Hz), 8.15
(d, 1H, ArH, J = 8.5 Hz), 8.00 (d, 1H, ArH,
J = 8.1 Hz), 7.80 (t, 1H, ArH, J = 6.9 Hz and
J = 14.4 Hz), 7.53 (m, 2H, ArH), 7.21 (d, 1H, ArH,
J = 7.2 Hz), 7.07 (m, 2H, ArH + NHCO), 6.70 (m, 3H,
ArH), 3.82 (s, 3H, OMe), 3.77 (s, 3H, OMe), 3.49 (q,
2H, CH2 J = 4.9 Hz and J = 5.4 Hz), 2.69 (m, 6H,
CH2), 2.35 (s, 3H, CH3); 13C NMR (75.5 MHz, CDCl3)
d: 168.9, 163.8, 149.4, 149.0, 148.9, 147.5, 140.0, 135.7,
132.7, 132.4, 131.3, 129.4, 129.1, 127.6, 127.3, 127.0,
126.7, 123.2, 121.4, 120.4, 119.8, 111.9, 111.3, 58.5,
55.8, 55.3, 41.6, 36.8, 32.9; MS: 513.22 [M+H]+. HRMS
[M]+: calcd for C30H32N4O4 512.2423; Found: 512.2431.
7.4.3.10. Preparation of 2-amino-N-{2-[[2-(3,4-meth-
oxyphenyl)ethyl](methyl)amino]hexyl} benzamide (21a).
Yield: 70%, yellow oil. IR (NaCl, cmÀ1); 3479
(NHCO); 3357 (NH2); 3051, 2928, 2853 (C–H); 1641
(CO); 1587 (NH2 deformation); 1516, 1465, (C@C
Ar); 1264 (tertiary amine); 737 (CH deformation
Ar); 1H NMR (300 MHz, CDCl3) d: 7.27 (m, 1H,
ArH), 7.15 (m, 1H, ArH), 6.71 (m, 3H, ArH), 6.59
(m, 2H, ArH), 6.20 (m, 1H, NHCO), 5.49 (br s, 2H,
NH2), 3.83 (s, 3H, OMe), 3.81 (s, 3H, OMe), 3.35
(q, 2H, CH2, 6.3 and 6.7 Hz), 2.70 (m, 2H, CH2),
2.57 (m, 2H, CH2), 2.39 (t, 2H, CH2, J = 7.2 Hz),
2.29 (s, 3H, CH3), 1.51 (m, 4H, CH2), 1.32 (m, 4H,
CH2); 13C NMR (75.5 MHz, CDCl3) d: 169.4, 148.9,
148.6, 147.4, 132.9, 132.1, 127.1, 120.6, 117.2, 116.5,
116.4, 112.1, 111.4, 59.7, 57.5, 55.9, 55.8, 42.1, 39.6,
33.2, 29.6, 27.1, 27.0, 26.9.
7.4.4.2. Preparation of N-[2-({[{[2-(3,4-methoxyphe-
nyl)ethyl](methyl)amino}propyl]amino}carbonyl)phenyl]-
quinoline-3-carboxamide (23a). To a solution of dichlo-
romethane (10 mL) containing 18a (0.184 mmol) were
added 3-quinoline acid chloride and DOWEX 1X8
(0.172 g, 100–200 mesh) to give a yellow solid after puri-
fication (0.0765 g, 79%). Mp 92–94 ꢁC; IR (NaCl, cmÀ1):
3054, 2925, 2851 (C–H); 1673, 1598 (CO); 1517, 1464,
1446 (C@C Ar); 1265 (tertiary amine); 738 (CH defor-
7.4.3.11. Preparation of 2-amino-N-[2-(6,7-methoxy-
3,4-dihydroisoquinolin-2(1H)-yl)hexyl] benzamide (21b).
Yield: 76%, gum yellow solid. IR (NaCl, cmÀ1): 3450
(NHCO); 3352 (NH2); 3052, 2927, 2853 (C–H); 1644
(CO); 1586 (NH2 deformation); 1517, 1465, (C@C
Ar); 1264 (tertiary amine); 738 (CH deformation
Ar). 1H NMR (300 MHz, CDCl3) d: 7.25 (m, 1H,
ArH), 7.10 (m, 1H, ArH), 6.54 (m, 2H, ArH), 6.47
(s, 1H, ArH), 6.34 (t, 1H, ArH, J = 5.4 Hz), 5.48 (s,
2H, NH2), 3.78 (s, 6H, OMe), 3.50 (s, 2H, CH2),
3.31 (m, 2H, CH2), 2.77 (t, 2H, CH2, J = 5.5 Hz),
2.65 (t, 2H, CH2, J = 5.5 Hz), 2.44 (t, 2H, CH2,
J = 7.9 Hz), 1.53 (m, 4H, CH2), 1.34 (m, 4H, CH2);
13C NMR (75.5 MHz, CDCl3) d: 169.4, 148.6, 147.5,
147.2, 132.0, 127.1, 126.6, 126.2, 117.2, 116.5, 111.5,
109.5, 58.2, 55.9, 55.7, 51.0, 39.6, 29.6, 28.6, 27.2,
27.0, 26.9.
1
mation Ar); H NMR (300 MHz, CDCl3) d: 13.09 (s,
1H, NHCO), 9.55 (d, 1H, ArH, J = 2.04 Hz), 8.92 (m,
1H, NHCO), 8.85 (m, 1H, ArH, J = 8.3Hz), 8.79 (m,
2H, ArH), 8.13 (d, 1H, ArH, J = 8.5 Hz), 7.97 (d, 1H,
ArH, J = 8.2 Hz), 7.77 (m, 1H, ArH), 7.53 (m, 2H,
ArH), 7.30 (d, 1H, ArH, J = 7.9 Hz), 7.03 (m, 1H,
ArH), 6.70 (m, 3H, ArH), 3.84 (s, 3H, OMe), 3.73 (s,
3H, OMe), 3.57 (m, 2H, CH2), 2.66 (m, 6H, CH2),
2.33 (s, 3H, CH3), 1.79 (m, 2H, CH2); 13C NMR
(75.5 MHz, CDCl3) d: 168.8, 163.8, 149.3, 149.1, 148.9,
147.5, 140.3, 135.6, 132.5, 132.2, 131.2, 129.3, 129.2,
127.6, 127.3, 126.9, 126.6, 122.9, 121.3, 120.4, 119.9,
111.8, 111.3, 60.1, 57.5, 55.8, 41.9, 40.9, 33.3, 24.6;
MS: 526.29 [M+]. HRMS [M+]: calcd for C31H34N4O4
526.2580; Found: 526.2584.
7.4.4. General procedure for the synthesis of compounds
22a–26a and 22b–26b. To a solution of dicloromethane
(10–20 mL) containing compounds 17a–21a or 17b–
21b (1 equiv) were added 3-quinoline acid chloride
(1.5 equiv) and DOWEX 1X8 (100–200 mesh) chloride
resin. The mixture was stirred for 30 h at room temper-
ature. The mixture was filtered and extracted with 1 N
NaOH (3· 10 mL). The organic phases were combined,
dried over anhydrous Na2SO4, filtered, and evaporated
under reduced pressure. Pure product was obtained by
flash chromatography on silica gel using a gradient con-
centration of solvent from 2% to 10% methanol (2 M
NH3) in dichloromethane.
7.4.4.3. Preparation of N-[2-({[{[2-(3,4-methoxyphe-
nyl)ethyl](methyl)amino} butyl]amino} carbonyl)phen-
yl]quinoline-3-carboxamide (24a). To a solution of
dichloromethane (10 mL) containing 19a (0.256 mmol)
were added 3-quinoline acid chloride and DOWEX
1x8 (0.180 g, 100–200 mesh) to give a sticky yellow oil
after purification (0.106 g, 77%); IR (NaCl, cmÀ1):
3396 (NHCO); 2924, 2849 (C–H); 1672, 1597 (CO);
1516, 1447, (C@C Ar); 1264 (tertiary amine); 737 (CH
1
deformation Ar); H NMR(300 MHz, CDCl3) d: 12.74
(s, 1H, NH), 9.52 (d, 1H, ArH, J = 2.04 Hz), 8.77 (m,
2H, ArH), 8.13 (m, 2H, ArH + NH), 7.97 (d, 1H,
ArH, J = 8.0 Hz), 7.76 (t, 1H, ArH, J = 8.4 Hz), 7.53

P. Labrie et al. / Bioorg. Med. Chem. 14 (2006) 7972–7987
7985
(m, 3H, ArH), 7.03 (t, 1H, ArH, J = 7.5 Hz), 6.72 (d,
1H, ArH, J = 8.5 Hz), 6.63 (d, 1H, ArH, J = 6.9 Hz),
3.80 (s, 3H, OMe), 3.77 (s, 3H, OMe), 3.42 (m, 2H,
CH2), 2.64 (m, 4H, CH2), 2.42 (t, 2H, CH2, J = 5.9 Hz),
2.22 (s, 3H, CH3), 1.62 (m, 4H, CH2); 13C NMR
(75.5 MHz, CDCl3) d: 169.3, 163.8, 149.4, 149.0, 148.9,
147.4, 139.8, 135.7, 132.6, 132.5, 131.3, 129.3, 129.2,
127.5, 127.4, 127.0, 126.9, 122.9, 121.4, 120.6, 120.5,
112.0, 111.4, 59.4, 56.9, 55.9, 55.8, 42.2, 40.1, 32.9, 27.3,
25.1; MS: 541.26 [M+H]+. HRMS [M+H]+: calcd for
C32H36N4O4 540.2736; Found: 541.2833.
7.4.4.6. Preparation of N-[2-({[2-(6,7-methoxy-3,4-
dihydroisoquinolin-2(1H)-yl)ethyl]amino}carbonyl) phen-
yl]quinoline-3-carboxamide (22b). To a solution of
dichloromethane (10 mL) containing 17b (0.279 mol)
were added 3-quinoline acid chloride and 0.262 g of
DOWEX 1x8 (100–200 mesh) to give a yellow solid after
purification (give 0.0556 g, 39%). Mp 192–194 ꢁC; IR
(NaCl, cmÀ1); 3055, 2924, 2853 (C–H); 1674, 1600
(CO); 1518, 1448, (C@C Ar); 1265 (tertiary amine);
739 (CH deformation Ar); 1H NMR (300 MHz, CDCl3)
d: 12.75 (s, 1H, NH), 9.56 (d, 1H, ArH, J = 2.2 Hz), 8.83
(m, 2H, ArH), 8.16 (d, 1H, ArH, J = 8.0 Hz), 8.00 (d,
1H, ArH, J = 8,0 Hz), 7.83 (m, 1H, ArH), 7.63 (t, 3H,
ArH, J = 7.0 Hz), 7.54 (t, 1H, ArH, J = 8.9 Hz ), 7.33
(m, 1H, NH), 7.11 (t, 1H, ArH, J = 7.2 Hz), 6.60
(s, 1H, ArH), 6.52 (s, 1H, ArH), 3.85 (s, 3H, OMe),
3.82 (s, 3H, OMe), 3.67 (m, 4H, CH2), 2.83
(m, 6H, CH2); 13C NMR (75.5 MHz, CDCl3) d: 169.2,
163.8, 149.4, 149.0, 147.9, 147.5, 139.9, 135.8, 132.9,
131.3, 129.4, 129.1, 127.6, 127.4, 127,0, 126.9,
125.8, 125.6, 123.3, 121.5, 120.0, 111.4, 109.5, 55.9,
55.8, 55.3, 50.6, 36.6, 28.4; MS: 510.3 [M+].
HRMS [M+]: calcd for C30H30N4O4 510.2267; Found:
510.2263.
7.4.4.4. Preparation of N-[2-({[{[2-(3,4-methoxyphe-
nyl)ethyl](methyl)amino}pentyl]amino} carbonyl)phenyl]-
quinoline-3-carboxamide (25a). To a solution of dichlo-
romethane (10 mL) containing 20a (0.167 mmol) were
added 3-quinoline acid chloride and DOWEX 1X8
(0.0742 g, 100–200 mesh) to give a sticky yellow oil after
purification (0.0413 g, 45%). IR (NaCl, cmÀ1); 2926,
2853 (C–H); 1672, 1599 (CO); 1516, 1448, (C@C Ar);
1264 (tertiary amine); 738 (CH deformation Ar); 1H
NMR (300 MHz, CDCl3) d: 12.60 (s, 1H, NH), 9.54
(d, 1H, ArH, J = 2.3 Hz), 8.77 (m, 2H, ArH + NH),
8.15 (d, 1H, ArH, J = 8.3 Hz), 7.98 (d, 1H, ArH,
J = 8.1 Hz), 7.77 (m, 1H, ArH), 7.53 (m, 3H, ArH),
7.10 (t, 1H, ArH, J = 7.1 Hz), 6.74 (m, 4H, ArH), 3.85
(s, 3H, OMe), 3.81 (s, 3H, OMe), 3.45 (q, 2H, CH2,
J = 6.1 and 7.0 Hz), 2.70 (m, 2H, CH2), 2.60 (m, 2H,
CH2), 2.41 (t, 2H, CH2, J = 7.0 Hz), 2.29 (s, 3H, CH3),
1.59 (m, 4H, CH2), 1.37 (m, 2H, CH2); 13C NMR
(75.5 MHz, CDCl3) d: 169.2, 163.8, 149.4, 149.0, 148.9,
147.4, 139.8, 135.8, 132.9, 132.7, 131.4, 129.4, 129.1,
127.5, 127.4, 126.9, 126.7, 123.2, 121.6, 120.6, 112.2,
111.4, 59.7, 57.3, 55.9, 55.8, 42.1, 40.0, 33.3, 29.2, 26.8,
24.7; MS: 555.27 [M+H]+. HRMS [M+H]+: calcd for
C33H38N4O4 554.2893; Found: 555.2981.
7.4.4.7. Preparation of N-[2-({[2-(6,7-methoxy-3,4-
dihydroisoquinolin-2(1H)-yl)propyl]amino} carbonyl)phenyl]-
quinoline-3-carboxamide (23b). To a solution of dichlo-
romethane (10 mL) containing 18b (0.252 mol) were
added 3-quinoline acid chloride and 0.157 g of DOWEX
1X8 (100–200 mesh) to give a yellow solid after purifica-
tion (0.0553 g, 42%); mp 158–159 ꢁC IR (NaCl, cmÀ1):
3057, 2923, 2853 (C–H); 1672, 1598 (CO); 1518, 1445,
(C@C Ar); 1265 (tertiary amine); 736 (CH deformation
1
Ar); NMR H (300 MHz, CDCl3) d: 13.19 (s, 1H, NH),
9.60 (br s, 1H, NH), 9.54 (d, 1H, ArH, J = 2.10 Hz),
8.77 (m, 2H, ArH), 8.15 (d, 1H, ArH, J = 8.5 Hz),
8.00 (d, 1H, ArH, J = 8.1 Hz), 7.76 (t, 1H, ArH,
J = 8.3 Hz), 7.59 (t, 1H, ArH, J = 7.7 Hz), 7.31 (t, 1H,
ArH, J = 7.9 Hz), 7.08 (d, 1H, ArH, J = 7.8 Hz), 6.63
(s, 1H, ArH), 6.53 (s, 1H, ArH), 6.28 (t, ArH,
J = 7.6 Hz), 3.89 (s, 3H, OMe), 3.80 (s, 3H, OMe),
3.66 (s, 2H, CH2), 3.60 (m, 2H, CH2), 2.80 (m, 6H,
CH2), 1.89 (m, 2H, CH2); NMR 13C (75.5 MHz, CDCl3)
d: 168.9, 163.9, 149.3, 149.2, 147.9, 147.6, 140.2, 135.6,
132.3, 131.2, 129.4, 129.2, 127.7, 127.3, 127.0, 126.9,
125.8, 125.7, 122.6, 121.1, 119.5, 111.5, 109.5, 58.6,
56.1, 55.9, 55.6, 51.4, 41.5, 28.8, 23.6; MS: 524.3 [M+].
HRMS [M+]: calcd for C31H32N4O4 524.2423; Found:
524.2430.
7.4.4.5. Preparation of N-[2-({[{[2-(3,4-methoxyphe-
nyl)ethyl](methyl)amino}hexyl]amino} carbonyl)phenyl]-
quinoline-3-carboxamide (26a). To a solution of dichlo-
romethane (10 mL) containing 21a (0.167 mmol) were
added 3-quinoline acid chloride and DOWEX 1X8
(0.157 g, 100–200 mesh) to give a yellow solid after
purification (0.0542 g, 57%). Mp 100–101 ꢁC; IR
(NaCl, cmÀ1): 3420 (NHCO); 3060, 2926, 2853 (C–
H); 1673, 1598 (CO); 1517, 1448, (C@C Ar); 1265 (ter-
tiary amine); 739 (CH deformation Ar); 1H NMR
(300 MHz, CDCl3) d: 12.59 (s, 1H, NHCO), 9.51 (d,
1H, ArH, J = 2.0 Hz), 8.75 (m, 2H, ArH), 8.13 (d,
1H, ArH, J = 8.4 Hz), 7.98 (d, 1H, ArH, J = 8.1 Hz),
7.79 (m, 1H, ArH), 7.59 (m, 2H, ArH), 7.49 (m, 1H,
ArH), 7.05 (m, 1H, ArH), 6.88 (m, 1H, NHCO), 6.69
(m, 3H, ArH), 3.82 (s, 3H, OMe), 3.80 (s, 3H, OMe),
3.43 (m, 2H, CH2), 2.70 (m, 2H, CH2), 2.60 (m, 2H,
CH2), 2.41 (t, 2H, CH2, J = 7.1 Hz), 2.29 (s, 3H,
CH3), 1.62 (m, 2H, CH2), 1.49 (m, 2H, CH2), 1.32
(m, 4H, CH2); 13C NMR (75.5 MHz, CDCl3) d:
169.2, 163.8, 149.4, 149.0, 148.9, 147.4, 139.7, 135.8,
132.7, 132.6, 131.4, 129.4, 129.2, 127.5, 127.4, 126.9,
126.8, 123.2, 121.6, 120.6, 112.1, 111.4, 59.5, 57.3,
55.9, 55.8, 41.9, 40.0, 33.1, 29.3, 26.9, 26.8; MS:
569.2873 [M+H]+. HRMS [M+H]+: calcd for
C34H40N4O4 568.3049; Found: 569.3130.
7.4.4.8. Preparation of N-[2-({[2-(6,7-methoxy-3,4-
dihydroisoquinolin-2(1H)-yl)butyl]amino} carbonyl)phen-
yl]quinoline-3-carboxamide (24b). To a solution of
dichloromethane (10 mL) containing 19b (0.119 mol)
were added 3-quinoline acid chloride and DOWEX
1X8 (0.111 g, 100–200 mesh) to give a sticky yellow oil
after purification (0.0507 g, 79%). IR (NaCl, cmÀ1);
2927, 2856 (C–H); 1600 (CO); 1519, 1448, (C@C Ar);
1265 (tertiary amine); 739 (CH deformation Ar); 1H
NMR (300 MHz, CDCl3) d: 12.80 (s, 1H, NH) 9.54 (d,
1H, ArH, J = 2.1 Hz), 8.75 (m, 2H, ArH), 8.32 (m,
1H, NH), 8.15 (d, 1H, ArH, J = 8.4 Hz), 8.00 (d, 1H,

7986
P. Labrie et al. / Bioorg. Med. Chem. 14 (2006) 7972–7987
ArH, J = 8,0 Hz), 7.78 (t, 1H, ArH, J = 7.9 Hz), 7.61 (t,
1H, ArH, J = 7.5Hz) 7.40 (t, 1H, ArH, J = 7.5 Hz), 7.26
(m, 1H, ArH), 6.51 (m, 2H, ArH), 6.40 (s, 1H, ArH),
3.84 (s, 3H, OMe), 3.80 (s, 3H, OMe), 3.50
(m, 4H, CH2), 2.74, (m, 4H, CH2), 2.56 (m, 2H, CH2),
1.78 (m, 4H, CH3); NMR 13C (75.5 MHz, CDCl3)
d: 169.2, 163.8, 149.4, 149.1, 147.8, 147.5, 139.6, 135.7,
132.2, 131.2, 129.4, 129.1, 127.6, 127.3, 127,0,
126.9, 125.8, 122.9, 121.3, 120.4, 111.4, 109.5, 56.9,
56.2, 55.9, 49.9, 39.9, 28.3, 26.8, 24.8; MS: 538.3 [M+].
HRMS [M+]: calcd for C32H34N4O4 538.2580; Found:
538.2590.
Acknowledgments
We thank Dr. W. T. Beck for providing us with cancer cell
lines CEM/VLB500 and CEM. This work was supported
by a grant from the Canadian Institute of Health Re-
search, and by scholarships from the Fonds de la recher-
´
che en sante du Quebec (P.L.) and the Natural Sciences
and Engineering Research Council of Canada (P.L.).
´
References and notes
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7.4.4.9. Preparation of N-[2-({[2-(6,7-methoxy-3,4-
dihydroisoquinolin-2(1H)-yl)pentyl]amino} carbonyl)phenyl]-
quinoline-3-carboxamide (25b). To a solution of dichlo-
romethane (10 mL) containing 20b (0.249 mol) were
added 3-quinoline acid chloride and 0.155 g of DOWEX
1X8 (100–200 mesh) to give a yellow solid after purifica-
tion (0.0751 g, 55%). Mp 87–89 ꢁC; IR (NaCl, cmÀ1);
3459 (NHCO); 3055, 2923, 2854 (C–H); 1672, 1598
(CO); 1519, 1446, (C@C Ar); 1267 (tertiary N); 740
1
(CH deformation Ar); NMR H (300 MHz, CDCl3) d:
12.58 (s, 1H, NH), 9.52 (d, 1H, ArH, J = 2.1 Hz), 8.74
(m, 2H, ArH), 8.13 (d, 1H, ArH, J = 8.5 Hz), 7.97 (d,
1H, ArH, J = 8.1 Hz), 7.78 (t, 1H, ArH, J = 7.9 Hz),
7.59 (t, 1H, ArH, J = 7.7 Hz), 7.49 (t, 1H, ArH,
J = 8.3 Hz), 7.44 (br s, 1H, NH), 6.98 (t, 1H, ArH,
J = 7.1 Hz), 6.53 (s, 1H, ArH), 6.46 (s, 1H, ArH), 3.80
(s, 3H, OMe), 3.78 (s, 3H, OMe), 3.50 (s, 2H, CH2),
3.44 (m, 2H, CH2), 2.76 (m, 2H, CH2), 2.65 (m, 2H,
CH2), 2.46 (t, 2H, J = 7.2 Hz), 1.60 (m, 4H, CH2), 1.40
(m, 2H, CH2); NMR 13C (75.5 MHz, CDCl3) d: 169.2,
163.9, 149.4, 148.9, 147.6, 147.3, 139.6, 135.7, 132.6,
131.4, 129.4, 129.2, 127.5, 127.4, 126.9, 126.7, 126.3,
126.1, 123.2, 121.5, 120.6, 111.4, 109.5, 57.9, 55.9,
55.8, 55.7, 50.9, 39.9, 29.3, 28.5, 26.7, 24.7; MS: 553.27
[M+H]+. HRMS [M+H]+: calcd for C33H36N4O4
552.2736; Found: 553.2815.
14. Miller, T. P.; Grogan, T. M.; Dalton, W. S.; Spier, C.
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7.4.5. Preparation of N-[2-({[2-(6,7-methoxy-3,4-dihydro-
isoquinolin-2(1H)-yl)hexyl]amino} carbonyl)phenyl]quino-
line-3-carboxamide (26b). To a solution of dichloromethane
(10 mL) containing 21b (0.324 mol) were added 3-quinoline
acid chloride and DOWEX 1X8 (0.303 g, 100–200 mesh) to
give a yellow solid after purification (0.113 g, 62%). Mp 78–
79 ꢁC;IR (NaCl, cmÀ1); 3055, 2927, 2856 (C–H); 1674, 1599
(CO); 1518, 1447, (C@C Ar); 1265 (tertiary amine); 739 (CH
deformation Ar). NMR 1H (300 MHz, CDCl3) d: 12.59 (s,
1H, NH), 9.49 (m, 1H, ArH), 8.70 (m, 2H, ArH), 8.10 (d,
1H, ArH, J = 8.4 Hz), 7.93 (d, 1H, ArH, J = 8.1 Hz), 7.75
(t, 1H, ArH, J = 7.0 Hz), 7.53 (m, 1H, ArH), 7.42 (t, 1H,
ArH, J = 7.6 Hz), 7.17 (m, 1H, NH), 6.97 (t, 1H, ArH,
J = 7.7 Hz), 6.50 (s, 1H, ArH), 6.43 (s, 1H, ArH), 3.76 (s,
3H, OMe), 3.74 (s, 3H, OMe), 3.42 (m, 4H, CH2), 2.73
(m, 2H, CH2), 2.60 (m, 2H, CH2), 2.40 (m, 2H, CH2), 1.40
(m, 8H, CH2); 13C NMR (75.5 MHz, CDCl3) d: 169.2,
163.8, 149.3, 148.9, 147.5, 147.2, 139.6, 135.7, 132.5,
131.4, 129.3, 127.4, 126.9, 126.7, 126.2, 123.2, 121.4,
120.6, 111.4, 109.5, 58.2, 55.9, 55.8, 55.7, 50.9, 40.1, 29.4,
28.6, 27.2, 27.1, 26.9; MS: 567.26 [M+H]+. HRMS
[M+H]+: calcd for C34H38N4O4 566.2893; Found:
567.2978.
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