Na+ currents in cardioprotection: Better to be late

Article abstract of DOI:10.1021/jm900296e

We report the discovery of a selective, potent inhibitor of the late current mediated by the cardiac isoform of the sodium channel (Na _V1.5). The compound, 3,4-dihydro-N-[(2S)-3-[(2-hydroxy-3- methylphenyl) thio]-2-methylpropyl]-2H-(3R)-1,5-ben

Full text of DOI:10.1021/jm900296e

J. Med. Chem. 2009, 52, 4149–4160 4149
DOI: 10.1021/jm900296e
Na+ Currents in Cardioprotection: Better to Be Late
†
Bruno Le Grand, Christophe Pignier, Robert Letienne, Francis Colpaert, Florence Cuisiat, Franc-oise Rolland,
†
†
§
‡
‡
ꢀ
Agnes Mas,‡ Maud Borras,‡ and Bernard Vacher*,‡
†Cardiovascular 2 Division and ‡Medicinal Chemistry 1 Division, Pierre Fabre Research Center, 17 avenue Jean Moulin, 81106 Castres Cedex,
France, and §Toulouse ISTMT 2, 3 rue des Satellites, 31400 Toulouse
Received March 9, 2009
We report the discovery of a selective, potent inhibitor of the late current mediated by the cardiac
isoform of the sodium channel (NaV1.5). The compound, 3,4-dihydro-N-[(2S)-3-[(2-hydroxy-3-methyl-
phenyl)thio]-2-methylpropyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2d) (F 15741), blocks the late
component of the Na+ currents and greatly reduces veratridine- or ischemia-induced contracture in
isolated tissue and whole heart. The cardioprotective action of 2d was further established in a model of
myocardial infarction in the pig in which 2d prevents ischemia-reperfusion damage after 60 min of
coronary occlusion and 48 h reperfusion. Under these experimental conditions, only 2d and cariporide
reduce infarct size. Remarkably, myocardial protection afforded by 2d occurs in the absence of
hemodynamic effects. These data expand the therapeutic potential of late INa blockers and suggest
that 2d could be useful in pathologies for which pharmacological treatments are not yet available.
Introduction
During ischemia, a fraction of the Na+ channels either fails
to inactivate or reopens inappropriately, amplifying the long-
We have previously described compound 1 (F 15845,
Figure 1) as the first selective, potent, and voltage-dependent
inhibitor of the late current mediated by the cardiac sodium
channel NaV1.5a.1 The compound reduces the mechanical
dysfunction induced by ischemia, hypoxia and toxins and
reverses electrocardiographic abnormalities following transi-
ent coronary artery occlusion in anesthetized rabbits and
dogs.2 In addition, the anti-ischemic activity of 1 is not
associated with any concomitant changes in blood pressure,
heart rate, inotropic status, or vascular tone. Consequently,
comedication with agents that mediate their therapeutic
benefits by a hemodynamic action remains an option (e.g.,
β-blockers, organic nitrates, etc.).3
The physiological importance of slow inactivation of vol-
tage gated sodium channels has been established in various
forms of inherited skeletal muscle,4 cardiac muscle,5 and
central and peripheral neuronal6 diseases or disorders in
man. Only recently, however, have the merits of a late INa
blocker (i.e., ranolazine)7-9 been acknowledged in the man-
agement of chronic angina10 and arrhythmias.11 In those
indications, however, the added benefits (if any) of a more
selective and more potent late INa blocker than ranolazine are
largely unknown.12 We now provide evidence that the action
of the selective late INa blocker 2d (F 15741, Figure 2) goes
beyond that of the classical of anti-ischemics, as 2d directly
protects the heart muscle against acute myocardial infarction,
offering thus novel prospects for therapy.13
+
lasting, voltage-dependent inward Na current referred to as
late INa. By preventing this influx of sodium, late INa inhibi-
tors attenuate cytosolic Na+ accumulation14 and, indirectly,
the uptake and build-up of Ca2+, which has devastating
consequences during both ischemia and reperfusion phases.15
In the course of our studies on a novel series of late INa
blockers,1 we noticed that structurally close, selective and, a
priori, equipotent veratridine-INa blockers can yet produce
divergent responses in models addressing different aspects
of myocardial ischemia (e.g., ECG abnormalities, rhythmic
disturbances or infarct size). Such context-dependent activity
suggests that the pharmacological profile of the compound is,
insomemanner, conditionedbythe“quality” of theinhibition
of the late current. On the basis of these observations, we
redirected our efforts and targeted the discovery of INa
blockers tailored for cardioprotection, an area where no
pharmacological intervention has proved successful in spite
of decades of research and a plethora of drugs investigated1.6
SAR mapping carried out on the structural archetype of
this series (Figure 3) reveals that, in addition to the propyl
chain,1 the aromatic tail is another site amenable to chemical
modulations. Prospecting in this direction yielded compounds
that exhibited the sought-after reinforced cardioprotective
properties. This effort culminated in the selection of the
development candidate 2d (Figure 2). Here, we focus on the
process that led to the discovery of 2d, touch upon some
intriguing mechanistic issues that are raised by this com-
pound, and illustrate its potential as cardioprotectant in
comparison with various reference agents (cf. Figure 1).
*To whom correspondence should be addressed. Phone: (+33) 56371
4222. Fax: (+33) 56371 4299. E-mail: bernard.vacher@pierre-fabre.
com.
a Abbreviations: INa, inward sodium current; NaV, voltage-gated
sodium channel; AP, action potential; TTX, tetrodotoxin; VIDC,
veratridine induced diastolic contracture; HEK, human embryonic
kidney; HP, holding potential; LPH, Langendorff-perfused heart;
CABG, coronary artery bypass graft surgery.
Chemistry
Two synthetic routes were used for the preparation of
compounds of the type 2. The first one, depicted in Scheme 1
r
2009 American Chemical Society
Published on Web 06/10/2009
pubs.acs.org/jmc

4150 Journal of Medicinal Chemistry, 2009, Vol. 52, No. 14
Le Grand et al.
Figure 1. Agents used as references.
inhibitors reduces the amplitude of this noninactivating cur-
rent.30 For compound selection, effects on the veratridine-
evoked INa currents were tested at a single concentration
(10 μM) but from two holding membrane potentials (HP) in
order to appreciate any HP dependence of the block. At both
HP values, the selective Na+ blocker TTX nearly abolishes
late INa, while the selective late INa blocker 1 markedly re-
presses it (Table 1). Removal of the etheral-oxygen atom (2a),
introducing a larger O-alkyl group (2b) or a substituent in
position 3 (2c) all have a negative impact on late INa inhibition
relative to 1. In contrast, a free OH in 2-position was clearly
beneficial, 2d being as efficacious as TTX and significantly
more so than 1 at suppressing late INa. When R1 = OH, re-
covery from slow inactivation varies inversely with the size of
R2; decreasing from Me (2d) to i-Pr (2f). Activity diminishes
upon moving the 2-OH group one carbon away from the ring
(2g vs 2d), indicating that mesomeric donation alone is insuffi-
cient to account for the phenolic-OH contribution. Switching
the phenol for aniline (R1 =NH2) is counterproductive and
remains so whatever the nature of R2 (cf. 2l, 2m, 2n). Likewise,
attempts at bioisosteric replacement of phenol (2s and 2t,
Table 2) were of no avail. Among the fused heterocycles, only
monoheteroatomic oxygenated ones (2o and 2p) have the
Figure 2. Structure of compound 2d (F 15741).
Figure 3. Structural archetype of the series.
and 2, involved alkylation of the appropriate arythiol or
protected arylthiol 3 with (S)-3-bromo-2-methylpropanol
under basic conditions to give the corresponding thi-
oether 4.1 Oxidation of the primary alcohol function in
4 under modified Swern conditions17 led to the sensitive
aldehyde 5, which was not isolated but carried forward into
the reductive amination step with 3-(R)-amino-benzoxathie-
pine618 to afford compound 2.19 Some compounds of the type
2 required an extra step (i.e., 2d-f, 2g-i, 2k-l) to access the
final material.
Alternatively (Scheme 3), reaction between (R)-3-amino-1,5-
benzoxathiepine 6, activated as its aluminum salt, and (R)-3-
hydroxy-2-methylpropionate, yielded the amide 7.20 Mesyla-
tion of the primary alcohol function in 7 then displacement of
the methanesulfonyloxy group with the appropriate arylthiol
(or disulfide precursor) produced the thioether 9. Subsequent
reduction of the amide delivered the target compound 2.21
Compound 2r was obtained from 2j by cyclization with
triethylorthoformate.22
Among the arylthiols (or disulfides) used in this work, a few
are commercially available (3a, 3b) and some are known: 2-
methoxy-3-methylbenzenethiol (3c),23 2-hydroxy-3-methyl-
benzenethiol (3d),24 2-hydroxy-3-ethylbenzenethiol (3e),25
2-hydroxy-3-isopropylbenzenethiol (3f),25 2-amino-3-methyl-
benzenethiol (3l),26 2-amino-3-methoxybenzenethiol (3m),27
2-amino-3-fluorobenzenethiol (3n),28 and 2,3-dihydrobenzo-
furan-7-thiol (3o);25 the others (i.e., 3h, 3i, 3p-q, and 3t) were
prepared as described in Supporting Information.
ability to reverse veratridine-induced late INa
.
Overall, a phenolic-OH at C-2 boosted late INa inhibition.
The role of the group at C-3 seems to be merely steric
inasmuch as electron-donating (2h) or -withdrawing groups
(2i) gave comparable results and size is tightly controlled (2d
vs 2f).
HP determines the state of the Na+ channels in the mem-
brane. At -110 mV, channels exist primarily in their rest-
ing state (available for opening), whereas at -90 mV, a
substantial proportion of them are inactivated. Because
blockade of late INa is generally more pronounced at -90
than at -110 mV, it seems that most compounds of this
series stabilize channels in their inactivated state, decreasing
their probability of opening. Of the derivatives listed in
Tables 1 and 2, compounds 2d and 2p stand out, matching
the inhibition attained with TTX. Further, 2d distinguishes
between late and peak INa currents (data not shown), which is
critical for the normal functioning of the heart.31 Note that, in
nonischemic cardiac cells, the peak INa carries most of the
current.1
To simplify matters, we ascribe the difference seen in the
activity patterns of analogues to a conformational selection
effect; assuming that different compounds prefer certain
protein conformation(s) among those that populate the
long-lasting, slow-inactivating state of the channel. This, in
turn, shouldinfluence the compoundsblocking characteristics
and, therefore, their pharmacological responses under patho-
logical conditions.
Results and Discussion
Exposure of HEK293 cells expressing hNaV1.5 R-subunits
to veratridine (40 μM) induces a 100-fold enhancement in the
late INa measured at 340 ms after initiation of the depolarizing
pulse, that is, long after recovery from the peak INa current.29
Typically, treatment of such a preparation with late INa

Article
Journal of Medicinal Chemistry, 2009, Vol. 52, No. 14 4151
Scheme 1. Synthesis of Compounds 2a-c, 2ga
a Reagents and conditions: (i) base, THF, or DMF; (ii) Swern oxidation; (iii) compound 6, NaBH(OAc)3, C2H4Cl2; (iv) LiAlH4, THF.
Scheme 2. Synthesis of Compounds 2d-f, 2h,i, 2k,l, 2o-q, 2s,ta
a Reagents and conditions: (i) Swern oxidation; (ii) compound 6, NaBH(OAc)3, C2H4Cl2; (iii) HCl (4 N), EtOH, 60 °C; (iv) HCl (12 N), MeOH, 60 °C;
(v) NH4OH, MeOH.
During ischemia, late INa contributes to ionic and mechan-
ical perturbations. When cardiac tissue (e.g., isolated left
atrium) is exposed to veratridine, the initial Na+ overload
evolves into a contraction-relaxation failure signaled by the
occurrence of a diastolic contracture. The latter is only slowly
reversible and resistant to inhibition by conventional anti-
ischemic agents, peak and nonselective sodium currents
blocking drugs such as class I antiarrhythmics.32 Now the
fact that several compounds in this series (cf. Tables 1 and 2)
can offset that contracture unequivocally dissociates their
mechanism from that of above-mentioned drugs.
Here, only inhibitions greater than 40% in both of the
veratridine-based procedures are taken as evidence for phar-
macodynamic action at late INa. On this basis, 2b and 2t
were discarded and the superiority of phenol over aniline
was confirmed. In the end, only a few compounds (2d, 2h,
2k, 2o, and 2q) achieve a level of efficacy in the range of that
of 1. In terms of stereochemistry-activity relationship,
aromatic and chain-modified compounds follow the same
pattern, thus we pursued structure-activity studies with a
single enantiomer.
Compounds were next probed in Langendorff-perfused
guinea pig hearts (abbreviated LPH hereafter).33 In this model
ofglobalischemia, weassessedthe abilityofthe compounds to
oppose the diastolic contracture that develops upon pro-
longed interruption of the coronary flow. Compounds in
Tables 1 and 2 can be separated in two groups: one encom-
passing derivatives the activities of which in VIDC and LPH
diverge (i.e., 2e, 2g, 2h, 2k, 2m, 2n, 2q-r, 2t), and another,
which is composed of derivatives efficacious in both models
(i.e, 2d, 2i, 2o, 2p). Among the latter derivatives, 2d and 2o
show even greater activity than 1 and do so without altering
baseline hemodynamic or contractile parameters.34
The lack of a direct correlation between veratridine- and
ischemia-promoted contracture can be understood if, in LPH,
mechanisms unrelated to late INa can also trigger a contrac-
ture or if veratridine evoked Na+ currents only imperfectly
35
.
mimic native late INa

4152 Journal of Medicinal Chemistry, 2009, Vol. 52, No. 14
Le Grand et al.
Overall, compound 2d met all the criteria defined at the
outset: it produces high anti-ischemic effects in different in
out the experiment, whereas circulation was absent in the
ischemic zone. In this study, the test compound was adminis-
teredintravenouslyasa bolus 10minbefore ischemia and then
as a continuous infusion for the duration of the occlusion
(60 min) and the first 30 min of reperfusion. Under those
conditions, cariporide38 significantly reduced infarct volume
to 6.8% ( 2.4, measured at the end of the reperfusion period
as compared with vehicle-treated controls (27.7% ( 3.0). This
result is interesting, bearing in mind that cariporide has shown
a beneficial effect in patients undergoing coronary artery
bypass graft surgery (CABG).39 The nonselective sodium
channel blocker lidocaine40 was not only ineffective but also
increased the incidence of ventricular fibrillation and death;
data in line again with clinical observations.41 The If inhibi-
tor ivabradine42 exerts no protection against cell death, con-
trary to reports in a low-flow, short-reperfusion pig model at a
dose equivalent to that used in this work.43 Remarkably,
compound 2d reduces infarct size to 3.5% ( 0.6, that is, 2d
salvages up to 90% of the area rendered ischemic that other-
wise would undergo necrosis (“area at risk”). Thus, hearts
exhibited hardly any detectable infarct at all.44 Further, at the
dose tested,45 treatment with 2d is not associated with mod-
ifications of blood pressure, heart rate, or oxygen consump-
tion, supporting a direct cardioprotective action.
vitro models and does so by selectively blocking late INa.
31 To
evaluate whether 2d also protects cardiomyocytes in vivo, we
examined its activity in a pig model of myocardial ischemia-
reperfusion (Figure 4);36 ischemia was induced by a 60 min left
circumflex coronary occlusion followed by 48 h reperfusion.
Such a time frame is particularly suited for the determination
of infarct size in a precise and robust manner.37 In the
nonischemic area, regional blood flows were similar through-
Scheme 3. Synthesis of Compounds 2j, 2m,n, and 2ra
These morphological data are backed by measures of
plasma levels of troponin I during reperfusion.46 That is, the
reduction of infarct size observed with 2d and cariporide is
accompanied by a significant decrease in the release of
troponin I in the circulation at 6 h postreperfusion (Fig-
ure 4).47 In contrast, compounds that do not preserve cardiac
cell viability also fail to lower troponin I levels.
a Reagents and conditions: (i) DIBAH, THF; (ii) CH3SO2Cl, NEt3,
THF; (iii) ArSH or (ArS)2, PPh3, K2CO3, THF, 60 °C; (iv) NaBH4, BF3.
Et2O, THF, reflux; (v) HC(OEt)3, EtOH, 80 °C.
Overall, the magnitude of the cardioprotection achieved
with 2d outperformed that of cariporide and other reference
Table 1. In Vitro SAR, Acyclic Groups R1 and R2
hNav1.5 channel blockade,a % inhibition at 10 μMe
diastolic contracture, % inhibition at 1 μMe
compd
R1
R2
HP -110 mVf
HP -90 mVg
rat atrium (VI)b,c
GP intact heartd
1
OMe
Me
H
H
53 ( 7
11 ( 5
41 ( 3
28 ( 7
73 ( 9
56 ( 11
38 ( 10
45 ( 6
35 ( 6
38 ( 6
11 ( 5
59 ( 4
26 ( 7
13 ( 10
11 ( 8
79 ( 2
70 ( 5
14 ( 9
54 ( 4
32 ( 6
87 ( 8
56 ( 10
46 ( 8
58 ( 7
46 ( 6
40 ( 8
19 ( 6
65 ( 5
26 ( 5
18 ( 11
19 ( 5
87 ( 3
74 ( 3
3 ( 12
33 ( 10
30 ( 14
93 ( 5
46 ( 7
20 ( 12
58 ( 5
77 ( 11
68 ( 3
25 ( 14
72 ( 12
49 ( 4
55 ( 7
56 ( 9
64 ( 10
65 ( 11
NDh
2a
2b
2c
2d
2e
2f
OEt
OMe
OH
H
NDh
Me
Me
Et
NDh
70 ( 6
13 ( 11
NDh
OH
OH
i-Pr
Me
OMe
F
2g
2h
2i
CH2OH
OH
OH
31 ( 18
46 ( 9
54 ( 13
NDh
2j
OH
OH
NH2
CONH2
Me
OMe
F
2k
2l
43 ( 17
35 ( 6
21 ( 8
33 ( 11
NDh
NH2
NH2
NH2
2m
2n
TTX
a Veratridine (40 μM) induced late sodium current in HEK cells expressing human Nav1.5 channels. b Veratridine (40 μM) induced (VI) diastolic
contracture in rat isolated left atrium. c A contracture of 40% was used as cutoff value. d Global ischemia induced contracture (50 min) in isolated
Langendorff-perfused guinea pig (GP) heart. e Compound concentration used in the experiment. f Percentage of inhibition of late sodium current elicited
at -30 mV from a holding potential (HP) of -110 mV. g Percentage of inhibition of late sodium current elicited at -30 mV from a holding potential of
-90 mV. h ND: not determined.

Article
Journal of Medicinal Chemistry, 2009, Vol. 52, No. 14 4153
Table 2. In Vitro SAR, Fused Heterocycles
hNav1.5 channel blockadea, % inhibition at 10 μMe
diastolic contracture, % inhibition at 1 μMe
compd
X
Y
Z
HP -110 mVf
HP -90 mVg
rat atrium (VI)b,c
GP intact heartd
2o
2p
O
O
CH2
CH
CH2
CH
CH
N
CH2
CH
O
47 ( 21
73 ( 7
32 ( 6
16 ( 13
8 ( 4
56 ( 21
82 ( 4
54 ( 5
26 ( 15
8 ( 2
85 ( 5
46 ( 10
71 ( 6
34 ( 9
5 ( 6
39 ( 13
0 ( 11
64 ( 10
72 ( 12
68 ( 14
22 ( 13
84 ( 6
NDh
2q
O
2r
2s
O
N
NH
NH
CH
CH
2t
34 ( 5
44 ( 11
73 ( 12
6 ( 9
Cariporide
TTX
79 ( 2
87 ( 3
NDh
a Veratridine (40 μM) induced late sodium current in HEK cells expressing human Nav1.5 channels. b Veratridine (40 μM) induced (VI) diastolic
contracture in rat isolated left atrium. c A contracture of 40% was used as cutoff value. d Global ischemia induced contracture (50 min) in isolated
Langendorff-perfused guinea pig (GP) heart. e Compoundconcentrationused in the experiment. f Percentageof inhibition of late sodium current elicited
at -30 mV from a holding potential (HP) of -110 mV. g Percentage of inhibition of late sodium current elicited at -30 mV from a holding potential of
-90 mV. h ND: not determined.
Figure 4. Upper panel: Histochemical determination of infarct size after 60 min ischemia and 48 h reperfusion in pigs. Infarct size is expressed
as the percentage of necrotic area versus area at risk (empty circles represent individual pigs). Lower panel: Plasma levels of troponin I at the
peak of concentration 6 h postreperfusion. Data are means ( SEM *P < 0.05, ***P < 0.001 versus vehicle.
agents.48 To be effective, 2d has to be applied before the
occlusion, which is consistent with its mode of action. Such
schedule considerations have to be qualified, however, first
because these experiments were conducted in otherwise
healthy pigs/hearts in which there should be no enhance-
ment in late Na+ current until the onset of ischemia,
second, because the compound cannot reach its target in the
ichemic zone during the ischemic period (no blood flow and

4154 Journal of Medicinal Chemistry, 2009, Vol. 52, No. 14
Le Grand et al.
no collateral circulation). Such conditions may be more
stringent than real-life ones where late INa is likely to be
constitutively heightened and perfusion in the ischemic terri-
tory may be facilitated by collateral networks.49 In any case,
2d may be more adapted for treatment of patients presenting a
risk of myocardial infarction (MI) and/or for the secondary
prevention of MI and/or for patients undergoing interven-
tions involving a period of ischemia.50
brine, dried over Na2SO4, filtered, and concentrated in vacuo.
The free base was purified by flash column chromatography.
General Method for the Reduction of Amide 9 into Amine 2. To
a suspension of NaBH4 (5 equiv) in THF (10 mL per 100 mg)
under inert atmosphere and cooled at 0 °C was added dropwise
boron trifluoroetherate (5 equiv) and the mixture stirred for
15 min. A solution of the amide 9 in THF (10 mL per g) was then
introduced and the mixture heated at reflux until completion.
Methanol was added slowly to the reaction mixture at room
temperature, and the mixture was concentrated under vacuo.
Further methanol and HCl (12 N) were added, and the mixture
refluxed for 1 h and then cooled to room temperature and stirred
overnight. The mixture was concentrated under vacuo and then
the residue taken up in water, neutralized with NaOH (20%),
and then extrated with dichloromethane. The combined organic
layer was washed with brine, dried over Na2SO4, filtered, and
concentrated in vacuo. The free base was purified by flash
column chromatography.
Conclusions
Derivatives of this novel family of late sodium channel
blockers display a surprising broad spectrum of activity,
stretching from ECG normalization to cell protection. This
indeed supports a major role for late INa in ischemia. Among
the compounds described here, 2d was instrumental in reveal-
ing the infarct-sparing potential of this class of inhibitors.
Further, contrary to conventional Na+ channel blockers and
most known late INa inhibitors, 2d exhibited NaV1.5 isoform
and late current selectivity, two features that are essential for
limiting off-target undesired effects. It emerges that 2d reg-
ulates late INa in such a way that cell protection overrides
other anti-ischemic properties, rendering its action comple-
mentary in scope to that of compound 1.
At the isolated organ level, we found that 2d counteracts the
contracture resulting from either impaired Na+ conductance
or an ischemic insult. Likewise, in a severe model of acute
myocardial ischemia-reperfusion in pig, 2d prevents postre-
perfusion injury whereas, among reference compounds, only
cariporide achieved a lesser degree of protection.
WebelievethatlateINa inhibitionisthe primarymechanism
by which 2d exerts its cardioprotective activity.31 However,
the site of action of 2d on NaV1.5 channels as well as the
biophysical characteristic(s) responsible for its innovative
profile are yet to be identified. Clearly, 2d may provide a
new opportunity in the treatment of conditions for which
medical needs are considerable like myocardial infarction,
cardiac surgery, cardiac insufficiency, and heart failure.
3,4-Dihydro-N-[(2S)-3-[(2-methylphenyl)thio]-2-methylpropyl]-
2H-(3R)-1,5-benzoxathiepin-3-amine (2a). The free base was
purified by flash column chromatography (silica gel, dichlor-
omethane/ethylacetate, 95:5). Crystallization of 2a as a mal-
eate salt gave a white powder mp = 144 °C. IR (KBr): 2969,
1
1604, 1571, 1447, 1354 ν cm-1. H NMR (DMSO-d6) δ 1.12
(d, J = 6.8 Hz, 3H), 2.15-2.18 (m, 1H), 2.31 (s, 3H), 2.87 (dd,
J =12.8, 8.0 Hz, 1H), 3.01 (dd, J=12.4, 7.2 Hz, 1H), 3.14-3.23
(m, 2H), 3.27-3.35 (m, 2H), 3.86 (s, 1H), 4.34 (d, J = 13.6 Hz,
1H), 4.45 (d, J=13.6 Hz, 1H), 6.04 (s, 2H), 6.91-6.93 (m, 3H),
7.21-7.29 (m, 3H), 7.34 (d, J=7.8 Hz, 1H), 7.39 (d, J=7.6 Hz,
1H). 13C NMR (DMSO-d6) δ 17.3 (CH3), 19.9 (CH3), 30.5
(CH), 31.0 (CH2), 36.1 (CH2), 49.2 (CH2), 57.7 (CH), 70.4
(CH2), 121.9 (CH), 124.1 (CH), 125.4 (CH), 126.2 (C), 126.6
(CH), 127.1 (CH), 128.8 (CH), 130.0 (CH), 131.5 (CH), 135.3
(C), 135.6 (2CH), 136.2 (C), 159.0 (C), 167.2 (2C); [R]D25 13.5°
(c 0.39, CH3OH). HPLC: eluting with acetonitrile/water/
KH2PO4, 400:600:6.8 g, tR 13.3 min. Anal. (C20H25NS2O
C4H4O4) C, H, N.
3
3,4-Dihydro-N-[(2S)-3-[(2-ethoxyphenyl)thio]-2-methylpropyl]-
2H-(3R)-1,5-benzoxathiepin-3-amine (2b). The free base was
purified by flash column chromatography (silica gel, dichlor-
omethane/ethylacetate, 96:4). Crystallization of 2b as a fuma-
rate salt gave a white powder mp = 124 °C. IR (KBr) ν 2980,
2933, 1577, 1473, 1441, 1239 cm-1. 1H NMR (DMSO-d6) δ 1.00
(d, J=6.8 Hz, 3H), 1.34 (t, J=6.8 Hz, 3H), 1.78 (m, J= 6.4 Hz,
1H), 2.58 (dd, J = 13.0, 6.4 Hz, 1H), 2.65-2.70 (m, 3H), 2.87
(dd, J=13.0, 6.8 Hz, 1H), 3.06-3.14 (m, 3H), 3.96 (dd, J= 12.4,
4.4 Hz, 1H), 4.05 (q, J=7.2 Hz, 2H), 4.17 (dd, J=12.4, 2.8 Hz,
1H), 6.61 (s, 2H), 6.91-7.00 (m, 4H), 7.11-7.18 (m, 2H),
7.26 (d, J = 6.8 Hz, 1H), 7.34 (d, J = 6.8 Hz, 1H). 13C NMR
(DMSO-d6) δ 14.6 (CH3), 17.6 (CH3), 32.6 (CH), 33.9
(CH2), 35.4 (CH2), 50.9 (CH2), 57.7 (CH), 63.7 (CH2), 73.4
(CH2), 111.7 (CH), 120.9 (CH), 121.9 (CH), 123.6 (CH), 125.0
(C), 126.3 (CH), 127.3 (C), 127.6 (CH), 128.4 (CH), 131.5 (CH),
134.3 (2CH), 155.6 (C), 159.6 (C), 166.6 (2C); [R]D25 0° (c 0.26,
CH3OH). HPLC: eluting with acetonitrile/water/KH2-
Experimental Section
Chemistry. Melting points were determined on a Buchi
530 melting point apparatus and were not corrected. 1H NMR
spectra were recorded on a Bruker Avance 400 spectrometer
operating at 400 MHz for 1H and 100 MHz for 13C. Chemical
shifts are reported in δ value (ppm) relative to an internal
standard of tetramethylsilane. Infrared (IR) spectra were
obtained on a Nicolet FT 510 P spectrophotometer. Microana-
lyses were obtained on a Fison EA 1108/CHN analyzer. Analy-
tical thin-layer chromatography were carried out on precoated
plates (silica gel, 60 F 254 Merck). Optical rotations were
measured on a Perkin-Elmer 241 model polarimeter. The purity
of final compounds were established by HPLC using Xbridge
8.5 μm, 4.6 mm  250 mm reverse phase column at a flow rate
of 1 mL/min. The compounds were detected at 220 nm. The
purity of final products was g95% unless otherwise noted.
Commercially available products were used without further
purification.
General Method for the Reductive Amination between Alde-
hyde 5 and (R)-3-amino-1,5-benzoxathiepine (6). A solution of
(R)-3-amino-1,5-benzoxathiepine 6 (1 equiv) in dry dichloro-
methane (10 mL per g) was added to the crude solution of the
aldehyde 5 cooled at -20 °C. The reaction mixture was stirred
for 20 min and then treated with sodium triacetoxyborohydride
(1.2 equiv) and stirring was continued for 2 h at -10 °C. The
reaction mixture was poured into a 10% aqueous sodium
hydrogen carbonate solution and extracted with dichloro-
methane. The combined organic layer was washed with water,
PO4, 400:600:6.8 g, tR 14.03 min. Anal. (C21H27NS2O2
C4H4O4) C, H, N.
3
3,4-Dihydro-N-[(2S)-3-[(2-methoxy-3-methylphenyl)thio]-2-me-
thylpropyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2c). The free
base was purified by flash column chromatography (silica gel,
dichloromethane/methanol, 99:1). Crystallization of 2c as a fu-
marate salt gave a white powder mp=107 °C. IR (KBr) ν 2933,
1572, 1473, 1458, 1353 cm-1. 1H NMR (DMSO-d6) δ 1.11 (d, J =
6.8 Hz, 3H), 2.17 (m, 1H), 2.30 (s, 3H), 2.84 (m, 1H), 3.00 (m, 1H),
3.11-3.28 (m, 4H), 3.28 (m, 1H), 3.71 (s, 3H), 3.84 (m, 1H), 4.37
(m, 1H), 4.47 (m, 1H), 6.05 (s, 2H), 7.03-7.08 (m, 4H), 7.16 (t, J =
4.6 Hz, 1H), 7.25 (t, J = 8.0 Hz, 1H), 7.39 (d, J = 7.2 Hz, 1H). 13
C
NMR (DMSO-d6) δ 15.6 (CH3), 17.3 (CH3), 30.4 (CH), 31.3
(CH2), 35.0 (CH2), 50.0 (CH2), 57.6 (CH), 59.1 (CH3), 70.4
(CH2), 121.8 (CH), 124.5 (CH), 125.0 (CH), 125.2 (CH), 126.4

Article
Journal of Medicinal Chemistry, 2009, Vol. 52, No. 14 4155
(C), 128.2 (CH), 128.7 (CH), 129.9 (C), 130.0 (C), 131.4 (CH),
135.6 (2CH), 155.1 (C), 159.2 (C), 167.1 (2C); [R]D25 12.4° (c 0.26,
CH3OH). HPLC: eluting with acetonitrile/water/KH2PO4,
(dd, J = 12.8, 7.6 Hz, 1H), 2.96 (dd, J = 12.0, 7.2 Hz, 1H), 3.10
(dd, J = 12.8, 5.6 Hz, 1H), 3.10-3.20 (m, 1H), 3.26 (d, J =
5.6 Hz, 2H), 3.79 (s, 1H), 4.32 (d, J = 12.0 Hz, 1H), 4.44 (d, J =
12.0 Hz, 1H), 4.68 (s, 2H), 4.85 (s, 1H), 6.04 (s, 2H), 7.03-7.08
(m, 3H), 7.17 (t, J = 7.6 Hz, 1H), 7.22-7.30 (m, 2H), 7.39 (d,
J = 7.2 Hz, 1H). 13C NMR (DMSO-d6) δ 17.2 (CH3), 19.2
(CH3), 30.6 (CH), 31.0 (CH2), 38.0 (CH2), 49.9 (CH2), 57.6
(CH), 58.5 (CH2), 70.4 (CH2), 121.9 (CH), 124.1 (CH), 126.2
(C), 127.3 (CH), 127.8 (CH), 128.3 (CH), 128.7 (CH), 131.4
(CH), 135.5 (2CH), 136.2 (C), 138.2 (C), 139.0 (C), 158.9 (C),
400:600:6.8 g, tR 13.39 min, 91.6%; Anal. (C21H27NS2O2
C4H4O4) C, H, N.
3
3,4-Dihydro-N-[(2S)-3-[(2-hydroxy-3-methylphenyl)thio]-2-
methylpropyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2d). The
free base was purified by flash column chromatography (silica
gel, dichloromethane). Crystallization of 2d as a fumarate salt
gave a white powder mp = 101-105 °C. IR (KBr) ν 3420, 2966,
25
1
1691, 1654, 1470, 1229 cm-1. H NMR (DMSO-d6) δ 0.98 (d,
167.1 (2C); [R]D 0° (c 0.22, CH3OH). HPLC: eluting with
acetonitrile/water/KH2PO4, 400:600:6.8 g, tR 6.39 min. Anal.
(C21H27NS2O2 C4H4O4) C, H, N.
J = 6.8 Hz, 3H), 1.75 (m, 1H), 2.16 (s, 3H), 2.54 (dd, J = 12.4,
6.0 Hz, 1H), 2.61-2.68 (m, 2H), 2.84 (dd, J = 12.0, 6.8 Hz, 1H),
2.95 (dd, J = 12.4, 7.2 Hz, 1H), 3.06-3.10 (m, 2H), 3.95 (dd, J =
12.0, 5.6 Hz, 1H), 4.16 (dd, J = 12.4, 3.2 Hz, 1H), 6.61 (s, 2H),
6.72 (t, J = 7.6 Hz, 1H), 6.98 (t, J = 7.6 Hz, 3H), 7.13-7.20 (m,
2H), 7.33 (d, J = 8.0 Hz, 1H). 13C NMR (DMSO-d6) δ 16.5
(CH3), 17.7 (CH3), 32.8 (CH), 34.2 (CH2), 37.9 (CH2), 50.9
(CH2), 57.6 (CH), 73.6 (CH2), 119.7 (CH), 121.8 (CH), 122.2
(C), 123.5 (CH), 124.7 (C), 127.3 (C), 128.3 (CH), 128.6 (CH),
129.2 (CH), 131.4 (CH), 134.6 (2CH), 153.7 (C), 159.6 (C),
3
3,4-Dihydro-N-[(2S)-3-[(2-hydroxy-3-methoxyphenyl)thio]-2-
methylpropyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2h). The free
base was purified by flash column chromatography (silica gel,
cyclohexane/acetone, 80:20). Crystallization of 2h as a maleate
salt gave a white powder mp = 112-114 °C. IR (KBr) ν 3517,
2973, 1603, 1572, 1473, 1354 cm-1. 1H NMR (DMSO-d6) δ 1.09
(d, J = 6.4 Hz, 3H), 2.09 (m, 1H), 2.79 (dd, J = 12.8, 7.6 Hz, 1H),
2.96 (dd, J = 12.0, 7.6 Hz, 1H), 3.04 (dd, J = 12.8, 5.6 Hz, 1H),
3.19 (dd, J = 12.4, 5.6 Hz, 1H), 3.27 (d, J = 5.6 Hz, 2H), 3.79 (s,
3H), 3.80 (s, 1H), 4.37 (d, J = 12.4 Hz, 1H), 4.44 (d, J = 12.4 Hz,
1H), 6.04 (s, 2H), 6.77 (t, J = 8.0 Hz, 1H), 6.86 (t, J = 7.6 Hz,
2H), 7.04-7.09 (m, 2H), 7.23 (t, J = 6.8 Hz, 1H), 7.39 (t, J =
6.8 Hz, 1H), 9.03 (s, 2H). 13C NMR (DMSO-d6) δ 17.2 (CH3),
30.6 (CH), 31.1 (CH2), 36.0 (CH2), 49.9 (CH2), 55.8 (CH3), 57.6
(CH), 70.4 (CH2), 110.0 (CH), 119.2 (CH), 121.4 (CH), 121.8
25
166.3 (2C); [R]D 6.6° (c 0.48, CH3OH). HPLC: eluting with
acetonitrile/water/KH2PO4, 400:600:6.8 g, tR 9.75 min. Anal.
(C20H25NS2O2 C4H4O4) C, H, N.
3
3,4-Dihydro-N-[(2S)-3-[(2-hydroxy-3-ethylphenyl)thio]-2-methyl-
propyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2e). The free base
was purified by flash column chromatography (silica gel,
cyclohexane/ethylacetate, 85:15). Crystallization of 2e as a
maleate salt gave a white powder mp = 120 °C. IR (KBr) ν 3405,
(CH), 121.9 (C), 124.8 (CH), 126.2 (C), 128.7 (CH), 131.4 (CH),
-
25
2966, 2934, 1572, 1446, 1353 cm-1 1H NMR (DMSO-d6)
.
135.5 (2CH), 144.5 (C), 147.4 (C), 158.9 (C), 167.1 (2C); [R]D
13.6° (c 0.27, CH3OH). HPLC: eluting with acetonitrile/water/
KH2PO4, 400:600:6.8 g, tR 8.33 min. Anal. (C20H25NS2O3
C4H4O4) C, H, N.
δ 1.09-1.14 (m, 6H), 2.09 (m, J = 6.0 Hz, 1H), 2.58 (q, J =
7.6 Hz, 2H), 2.76 (dd, J = 12.8, 7.6 Hz, 1H), 2.95-3.02 (m, 2H),
3.19 (dd, J = 12.0, 6.0 Hz, 1H), 3.28 (d, J = 5.2, 2H), 3.81 (s,
1H), 4.36 (d, J = 13.2 Hz, 1H), 4.45 (d, J = 12.0 Hz, 1H), 6.05
(s, 2H), 6.80 (t, J = 7.6 Hz, 1H), 7.00-7.09 (m, 3H), 7.17 (d, J =
7.6 Hz, 1H), 7.25 (t, J = 7.6 Hz, 1H), 7.25 (d, J = 7.6 Hz, 1H),
8.58 (s, 2H). 13C NMR (DMSO-d6) δ 14.1 (CH3), 17.2 (CH3),
22.9 (CH2), 30.6 (CH), 31.1 (CH2), 37.4 (CH2), 49.8 (CH2), 57.6
(CH), 70.4 (CH2), 120.1 (CH), 121.8 (C), 121.9 (CH), 124.1
(CH), 126.7 (C), 127.6 (CH), 128.7 (CH), 128.8 (CH), 130.9 (C),
3
3,4-Dihydro-N-[(2S)-3-[(2-hydroxy-3-fluorophenyl)thio]-2-
methylpropyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2i). The free
base was purified by flash column chromatography (silica gel,
cyclohexane/acetone, 85:15). Crystallization of 2i as a maleate
salt gave a white powder mp = 110-112 °C. IR (KBr) ν 3398,
2962, 2702, 1601, 1475, 1354 cm-1. 1H NMR (DMSO-d6) δ 1.10
(d, J = 6.4 Hz, 3H), 2.12(m, 1H), 2.83 (dd, J = 12.8, 7.6 Hz, 1H),
2.98 (dd, J = 12.4, 7.6 Hz, 1H), 3.10 (dd, J = 12.8, 5.4 Hz, 1H),
3.19 (dd, J = 12.4, 6.0 Hz, 1H), 3.28 (d, J = 5.2 Hz, 2H), 3.83 (s,
1H), 4.37 (d, J = 12.0 Hz, 1H), 4.46 (d, J = 12.0 Hz, 1H), 6.04 (s,
2H), 6.80-6.85 (m, 1H), 7.03-7.09 (m, 4H), 7.24 (t, J = 7.6 Hz,
1H), 7.40 (d, J = 6.8 Hz, 1H). 13C NMR (DMSO-d6) δ 17.2
(CH3), 30.6 (CH), 31.1 (CH2), 35.9 (CH2), 49.9 (CH2), 57.6
(CH), 70.4 (CH2), 113.5 (CH, J = 19 Hz), 119.5 (CH), 121.9
(CH), 124.1 (CH), 124.5 (CH), 125.9 (C, J = 54 Hz), 128.3 (CH),
131.4 (CH), 135.5 (2CH), 142.6 (C), 150.0 (C), 152.4 (C), 158.9
(C), 167.1 (2C); [R]D25 -13.6° (c 0.27, CH3OH). HPLC: eluting
with acetonitrile/water/KH2PO4, 400:600:6.8 g, tR 8.33 min.
25
131.4 (CH), 135.5 (2CH), 153.1 (C), 158.9 (C), 167.1 (2C); [R]D
1.4° (c 0.23, CH3OH). HPLC: eluting with acetonitrile/water/
KH2PO4, 400:600:6.8 g, tR 9.75 min. Anal. (C21H27NS2O2
C4H4O4) C, H, N.
3
3,4-Dihydro-N-[(2S)-3-[(2-hydroxy-3-i-propylphenyl)thio]-2-
methylpropyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2f). The free
base was purified by flash column chromatography (silica gel,
cyclohexane/ethylacetate, 80:20). Crystallization of 2f as a mal-
eate salt gave a white powder mp = 126 °C. IR (KBr) ν 3410,
2962, 1572, 1473, 1438, 1353 cm-1. 1H NMR (DMSO-d6) δ 1.09
(d, J = 6.4 Hz, 3H), 1.15 (d, J = 6.8 Hz, 6H), 2.09 (m, J = 6.0 Hz,
1H), 2.75 (dd, J = 12.4, 7.6 Hz, 1H), 2.96-3.00 (m, 2H), 3.16 (m,
1H), 3.20-3.27 (m, 3H), 3.78 (s, 1H), 4.35 (d, J = 13.2 Hz, 1H),
4.44 (d, J = 13.2 Hz, 1H), 6.04 (s, 2H), 6.84 (t, J = 7.6 Hz, 1H),
7.06-7.08 (m, 3H), 7.17 (d, J = 7.6 Hz, 1H), 7.24 (t, J = 7.6 Hz,
1H), 7.39 (d, J = 7.6 Hz, 1H), 8.52 (s, 2H). 13C NMR (DMSO-d6)
δ 17.2 (CH3), 22.6 (2CH3), 26.4 (CH), 30.7 (CH), 31.1 (CH2),
37.7(CH2), 49.8 (CH2), 57.6(CH), 70.4(CH2), 120.3(CH), 121.8
(CH), 122.1 (C), 124.0 (CH), 124.8 (CH), 124.9 (C), 126.2 (C),
128.6 (CH), 128.7 (CH), 131.4 (CH), 135.5 (2CH), 152.5 (C),
158.6 (C), 167.1 (2C); [R]D25 4.1° (c 0.24, CH3OH). HPLC: eluting
with acetonitrile/water/KH2PO4, 400:600:6.8 g, tR 21.23 min.
Anal. (C19H22FNS2O2 C4H4O4) C, H, N.
3
3,4-Dihydro-N-[(2S)-3-[(2-hydroxy-3-aminophenyl)thio]-2-
methylpropyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2j). The free
base was purified by flash column chromatography (silica gel,
dichloromethane/ethylacetate, 90:10). Crystallization of 2j as a
maleate salt gave a white powder mp = 125 °C. IR (KBr) ν 3357,
2962, 2714, 1615, 1475, 1455, 1350 cm-1. 1H NMR (DMSO-d6) δ
1.08 (d, J = 6.4 Hz, 3H), 2.06 (m, 1H), 2.71 (dd, J = 12.8, 7.6 Hz,
1H), 2.94-2.97 (m, 2H), 3.17 (dd, J = 12.4, 5.6 Hz, 1H), 3.28 (d,
J = 5.2 Hz, 2H), 3.80 (s, 1H), 4.35 (d, J = 12.8 Hz, 1H), 4.44 (d,
J = 13.2 Hz, 1H), 6.05 (s, 2H), 6.54-6.63 (m, 3H), 7.03-7.09 (m,
2H), 7.24 (t, J = 7.2 Hz, 1H), 7.39 (d, J = 7.6 Hz, 1H). 13C NMR
(DMSO-d6) δ 17.2 (CH3), 30.7 (CH), 31.1 (CH2), 37.3 (CH2),
49.9 (CH2), 57.6 (CH), 70.4 (CH2), 113.6 (CH), 118.6 (CH),
120.5 (CH), 121.7 (C), 121.9 (CH), 124.1 (CH), 126.1 (C), 128.7
(CH), 131.4 (CH), 135.5 (2CH), 137.5 (C), 142.3 (C), 158.9 (C),
Anal. (C22H29NS2O2 C4H4O4) C, H, N.
3
3,4-Dihydro-N-[(2S)-3-[(2-hydroxymethyl-3-methylphenyl)
thio]-2-methylpropyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2g).
The free base was purified by flash column chromatography
(silica gel, dichloromethane/methanol, 98:2). Crystallization of
2g as a maleate salt gave a white powder mp = 102-104 °C. IR
(KBr) ν 3405, 1572, 1474, 1447, 1353 cm-1. 1H NMR (DMSO-
d6) δ 1.09 (d, J = 6.8 Hz, 3H), 2.08 (m, 1H), 2.38 (s, 3H), 2.85
25
167.1 (2C); [R]D 2.1° (c 0.29, CH3OH). HPLC: eluting with
acetonitrile/water/KH2PO4, 400:600:6.8 g, tR 8.78 min. Anal.
(C19H24NS2O2 C4H4O4) C, H, N.
3

4156 Journal of Medicinal Chemistry, 2009, Vol. 52, No. 14
Le Grand et al.
3,4-Dihydro-N-[(2S)-3-[(2-hydroxy-3-carboxamidophenyl)
thio]-2-methylpropyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2k).
The free base was purified by flash column chromatography
(silica gel, cyclohexane/acetone, 70:30). Crystallization of 2k as
a maleate salt gave a white powder mp = 154 °C. IR (KBr) ν
3474, 3347, 2978, 2822, 1708, 1654, 1472, 1414, 1356 cm-1. 1H
NMR (DMSO-d6) δ 1.09 (d, J = 6.4 Hz, 3H), 2.14 (m, 1H), 2.84
(dd, J = 12.8, 7.6 Hz, 1H), 3.00 (dd, J = 12.8, 7.6 Hz, 1H), 3.12
(dd, J = 13.2, 5.6 Hz, 1H), 3.21 (dd, J = 12.4, 5.6 Hz, 1H), 3.29
(d, J = 5.2 Hz, 2H), 3.80 (s, 1H), 4.35 (d, J = 13.6 Hz, 1H), 4.46
(d, J = 13.6 Hz, 1H), 6.05 (s, 2H), 6.88 (t, J = 8.0 Hz, 1H), 7.04-
7.09 (m, 2H), 7.25 (t, J = 8.6 Hz, 1H), 7.39 (d, J = 7.6 Hz, 1H),
7.46 (d, J = 7.6 Hz, 1H), 7.73 (d, J = 8.0 Hz, 1H), 8.08 (s, 1H),
8.56 (s, 1H). 13C NMR (DMSO-d6) δ 17.2 (CH3), 30.5 (CH),
31.1 (CH2), 35.3 (CH2), 49.9 (CH2), 57.6 (CH), 70.4 (CH2),
113.2 (C), 118.3 (CH), 121.8 (CH), 124.1 (CH), 124.2 (C), 125.2
(CH), 126.2 (C), 128.7 (CH), 131.4 (CH), 133.0 (CH), 135.5
(s, 2H), 6.53-6.58 (m, 1H), 7.00-7.09 (m, 3H), 7.15 (d, J =
7.6 Hz, 1H), 7.22-7.31 (m 1H), 7.38 (d, J = 7.6 Hz, 1H). 13C
NMR (DMSO-d6) δ 17.1 (CH3), 30.9 (CH), 31.0 (CH2), 38.8
(CH2), 49.7 (CH2), 57.6 (CH), 70.4 (CH2), 114.6 (J = 19 Hz,
CH), 115.6 (J = 8 Hz, CH), 118.7 (C), 121.8 (CH), 124.0 (CH),
126.1 (C), 128.7 (CH), 129.6 (CH), 131.4 (CH), 135.6 (2CH),
137.1 (C), 150.3 (J = 238 Hz, C), 158.9 (C), 167.1 (2C); [R]D25 3.8°
(c 0.29, CH3OH). HPLC: eluting with acetonitrile/water/
KH2PO4, 400:600:6.8 g, tR 7.83 min. Anal. (C19H23FN2S2O
C4H4O4) C, H, N.
3
3,4-Dihydro-N-[(2S)-3-[2,3-dihydrobenzofuran-7-thio]-2-
methylpropyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2o). The free
base was purified by flash column chromatography (silica gel,
dichloromethane/methanol, 99:1). Crystallization of 2o as a
maleate salt gave a white powder mp = 117-119 °C. IR (KBr)
1
ν 2966, 1573, 1448, 1382, 1354 cm-1. H NMR (DMSO-d6) δ
1.07 (d, J = 6.8 Hz, 3H), 2.08 (m, 1H), 2.83 (dd, J = 13.2, 7.6 Hz,
1H), 2.95 (dd, J = 12.4, 7.2 Hz, 1H), 3.10 (dd, J = 12.8, 5.6 Hz,
1H), 3.20 (t, J = 8.8 Hz, 3H), 3.26 (d, J = 5.6 Hz, 2H), 3.81 (s,
1H), 4.36 (d, J = 12.0 Hz, 1H), 4.45 (d, J = 13.6 Hz, 1H), 4.56 (t,
J = 8.8 Hz, 2H), 6.05 (s, 2H), 6.82 (t, J = 7.6 Hz, 1H), 7.03-7.14
25
(2CH), 158.9 (C), 159.5 (C), 167.1 (2C), 172.6 (C); [R]D 4.9°
(c 0.34, CH3OH). HPLC: eluting with acetonitrile/water/
KH2PO4, 400:600:6.8 g, tR 22.10 min. Anal. (C20H24N2S2O3
C4H4O4) C, H, N.
3
(m, 4H), 7.25 (t, J = 8.0 Hz, 1H), 7.40 (d, J = 7.6 Hz, 1H). 13
C
3,4-Dihydro-N-[(2S)-3-[(2-amino-3-methylphenyl)thio]-2-
methylpropyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2l). The free
base was purified by flash column chromatography (silica gel,
dichloromethane/ethylacetate, 95:5). Crystallization of 2l
as a maleate salt gave a white powder mp = 135 °C. IR (KBr)
NMR (DMSO-d6) δ 17.1 (CH3), 29.3 (CH2), 30.8 (CH), 31.1
(CH2), 36.3 (CH2), 49.7 (CH2), 57.5 (CH), 70.4 (CH2), 70.9
(CH2), 115.5 (C), 120.9 (CH), 121.8 (CH), 123.6 (CH), 124.1
(CH), 126.2 (C), 127.3 (C), 128.7 (2CH), 131.4 (CH), 135.6
25
ν 3470, 3356, 1605, 1573, 1470 cm-1 1H NMR (DMSO-d6)
.
(2CH), 158.3 (C), 158.9 (C), 167.1 (2C); [R]D -5.6° (c 0.49,
CH3OH). HPLC: eluting with acetonitrile/water/KH2PO4,
δ 1.08 (d, J = 6.4 Hz, 3H), 2.02 (m, 1H), 2.11 (s, 3H), 2.65
(dd, J = 13.2, 8.0 Hz, 1H), 2.89-2.97 (m, 2H), 3.14 (m, 1H), 3.26
(d, J = 4.0 Hz, 2H), 3.78 (s, 1H), 4.36 (d, J = 12.8 Hz, 1H), 4.44
(d, J = 13.2 Hz, 1H), 6.04 (s, 2H), 6.50 (t, J = 7.4Hz, 1H), 6.95 (d,
J = 7.2 Hz, 1H), 7.03-7.08 (m, 2H), 7.18 (d, J = 7.4 Hz, 1H),
7.24 (t, J = 7.2 Hz, 1H), 7.38 (d, J = 7.0 Hz, 1H). 13C NMR
(DMSO-d6) δ 17.1 (CH3), 18.1 (CH3), 30.9 (CH), 31.0 (CH2),
38.5 (CH2), 49.8 (CH2), 57.6 (CH), 70.4 (CH2), 115.8 (C), 116.4
(CH), 121.8 (CH), 121.9 (C), 124.0 (CH), 126.1 (C), 128.7 (CH),
130.3 (C), 131.4 (CH), 132.4 (CH), 135.5 (2CH), 146.9 (C), 158.9
(C), 167.1 (2C); [R]D25 14.2° (c 0.09, CH3OH). HPLC:elutingwith
acetonitrile/water/KH2PO4, 400:600:6.8 g, tR 8.45 min. Anal.
400:600:6.8 g, tR 10.18 min. Anal. (C21H25NS2O2 C4H4O4) C,
H, N.
3
3,4-Dihydro-N-[(2S)-3-[benzofuran-7-thio]-2-methylpropyl]-
2H-(3R)-1,5-benzoxathiepin-3-amine (2p). The free base was
purified by flash column chromatography (silica gel, dichloro-
methane/ethylacetate, 95:5). Crystallization of 2p as a maleate
salt gave a white powder mp = 112 °C. IR (KBr) ν 2966, 1572,
1
1473, 1448, 1353 cm-1. H NMR (DMSO-d6) δ 1.11 (d, J =
6.8 Hz, 3H), 2.13 (m, 1H), 2.97-3.04 (m, 2H), 3.20 (dd, J = 12.4,
6.0 Hz, 1H), 3.26-3.33(m, 3H), 3.81 (s, 1H), 4.34 (d, J=12.0 Hz,
1H), 4.44 (d, J=13.2 Hz, 1H), 6.05 (s, 2H), 6.98 (d, J = 2.0 Hz,
1H), 7.02-7.09 (m, 2H), 7.22-7.28 (m, 2H), 7.36-7.41 (m, 2H),
7.57 (d, J = 7.6 Hz, 1H), 8.04 (d, J = 2.0 Hz, 1H). 13C NMR
(DMSO-d6) δ 17.1 (CH3), 30.9 (CH2), 31.1 (CH), 38.8 (CH2),
49.7 (CH2), 57.5 (CH), 70.4 (CH2), 70.9 (CH2), 107.1 (CH),
118.5 (C), 119.6 (CH), 121.8 (CH), 123.6 (CH), 124.1 (CH), 124.7
(CH), 126.2 (C), 127.3 (C), 128.7 (CH), 131.4 (CH), 135.6 (2CH),
146.1 (CH), 152.9 (C), 158.9 (C), 167.1 (2C); [R]D25 -9.4° (c 0.28,
CH3OH). HPLC: eluting with acetonitrile/water/KH2PO4,
(C20H26N2S2O C4H4O4) C, H, N.
3
3,4-Dihydro-N-[(2S)-3-[(2-amino-3-methoxyphenyl)thio]-2-
methylpropyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2m). The
free base was purified by flash column chromatography (silica
gel, dichloromethane/ethylacetate, 95:5). Crystallization of 2m
as a maleate salt gave a white powder mp = 120 °C. IR (KBr) ν
3470, 3351, 2996, 2966, 1602, 1562, 1474 cm-1 1H NMR
.
(DMSO-d6) δ 1.08 (d, J = 6.4 Hz, 3H), 2.02 (m, 1H), 2.65 (dd,
J =13.2, 8.0 Hz, 1H), 2.91 (dd, J =12.8, 5.2 Hz, 2H), 3.12 (m,
1H), 3.25 (d, J = 4.8 Hz, 2H), 3.77 (s, 4H), 4.36 (d, J =
12.8 Hz, 1H), 4.43 (d, J =12.8 Hz, 1H), 6.04 (s, 2H), 6.56 (t,
J = 8.0 Hz, 1H), 6.81 (d, J = 7.6 Hz, 1H), 6.95 (d, J = 7.6 Hz,
1H), 7.03-7.08 (m, 2H), 7.24 (t, J =7.2 Hz, 1H), 7.38 (d, J = 7.6
Hz, 1H). 13C NMR (DMSO-d6) δ 17.1 (CH3), 30.9 (CH), 31.0
(CH2), 38.2 (CH2), 49.7 (CH2), 55.5 (CH3), 57.6 (CH), 70.4
(CH2), 110.2 (CH), 115.8 (C), 116.1 (CH), 121.8 (CH), 124.0
(CH), 126.1 (CH), 128.7 (CH), 131.4 (CH), 135.6 (2CH), 138.4
400:600:6.8 g, tR 11.68 min. Anal. (C21H23NS2O2 C4H4O4)
C, H, N.
3
3,4-Dihydro-N-[(2S)-3-[2,3-benzenedioxole-7-thio]-2-methyl-
propyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2q). The free base
was purified by flash column chromatography (silica gel, di-
chloromethane/ethylacetate, 95:5). Crystallization of 2q as a
maleate salt gave a white powder mp = 126 °C.IR (KBr) ν 2966,
1
1577, 1473, 1441, 1352 cm-1. H NMR (DMSO-d6) δ 1.07 (d,
J = 6.4 Hz, 3H), 2.09 (m, 1H), 2.86 (dd, J = 12.8, 7.6 Hz, 1H),
2.97 (m, 1H), 3.16 (dd, J = 12.8, 5.2 Hz, 1H), 3.25-3.32 (s, 3H),
3.82 (s, 1H), 4.35 (d, J = 12.8 Hz, 1H), 4.45 (d, J = 12.0 Hz, 1H),
6.04 (s, 2H), 6.05 (s, 2H), 6.83-6.91 (m, 3H), 7.03-7.09 (m, 2H),
7.25 (t, J = 8.0 Hz, 1H), 7.39 (d, J = 7.2 Hz, 1H). 13C NMR
(DMSO-d6) δ 16.9 (CH3), 30.9 (CH2), 31.1 (CH), 38.8 (CH2),
49.7 (CH2), 57.5 (CH), 70.4 (CH2), 100.9 (CH2), 107.1 (CH),
115.4 (C), 121.9 (CH), 122.2 (CH), 123.2 (CH), 124.1 (CH),
126.2 (C), 128.7 (CH), 131.5 (CH), 135.6 (2CH), 146.2 (C),
146.5 (C), 158.9 (C), 167.1 (2C); [R]D25 -1.6° (c 0.12, CH3OH).
HPLC: eluting with acetonitrile/water/KH2PO4, 400:600:6.8 g,
25
(2C), 146.3 (C), 158.9 (C), 167.1 (2C); [R]D 7.5° (c 0.23,
CH3OH). HPLC: eluting with acetonitrile/water/KH2PO4,
400:600:6.8 g, tR 7.80 min. Anal. (C20H26N2S2O2 C4H4O4)
C, H, N.
3
3,4-Dihydro-N-[(2S)-3-[(2-amino-3-fluorophenyl)thio]-2-
methylpropyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2n). The free
base was purified by flash column chromatography (silica
gel, cyclohexane/acetone, 80:20). Crystallization of 2n as a
maleate salt gave a white powder mp=126 °C. IR (KBr) ν
3457, 3349, 2966, 1616, 1589, 1474, 1353 cm-1 1H NMR
.
t
R 9.60 min. Anal. (C20H23NS2O3 C4H4O4) C, H, N.
3,4-Dihydro-N-[(2S)-3-[1,3-benzisoxazole-7-thio]-2-methyl-
(DMSO-d6) δ 1.08 (d, J = 6.4 Hz, 3H), 2.02 (m, 1H), 2.73 (dd,
J = 12.8, 7.6 Hz, 1H), 2.92-2.99 (m, 2H), 3.15 (dd, J = 12.4,
5.6 Hz, 1H), 3.26 (d, J = 5.2 Hz, 2H), 3.80 (s, 1H), 4.36 (d,
J = 12.0 Hz, 1H), 4.45 (d, J = 12.0 Hz, 1H), 5.29 (s, 2H), 6.05
3
propyl]-2H-(3R)-1,5-benzoxathiepin-3-amine (2r). Compound 2r
is not purified but crystallized directly as a maleate salt to give a

Article
Journal of Medicinal Chemistry, 2009, Vol. 52, No. 14 4157
white powder mp=112 °C. IR (KBr) ν 2966, 2819, 1603, 1573,
1473, 1353 cm-1. 1H NMR (DMSO-d6)δ1.11(d, J=6.4 Hz, 3H),
2.12 (m, 1H), 2.97-3.06 (m, 2H), 2.97 (m, 1H), 3.19 (dd, J =
12.4, 5.6 Hz, 1H), 3.26 (d, J = 5.0 Hz, 2H), 3.35 (dd, J = 13.2,
5.6 Hz, 1H), 3.81 (s, 1H), 4.34 (d, J = 12.4 Hz, 1H), 4.53 (d, J=
10.8 Hz, 1H), 6.04 (s, 2H), 7.04-7.09 (m, 2H), 7.25 (t, J =
7.2 Hz, 1H), 7.41 (t, J = 8.0 Hz, 2H), 7.49 (d, J = 7.6 Hz, 1H),
7.70 (d, J = 7.6 Hz, 1H), 8.81 (s, 1H). 13C NMR (DMSO-d6) δ
16.9 (CH3), 30.9 (CH2), 31.2 (CH), 36.4 (CH2), 49.7 (CH2), 57.6
(CH), 70.4 (CH2), 118.2 (CH), 118.6 (C), 121.9 (CH), 123.7 (C),
124.1 (CH), 125.3 (CH), 126.0 (C), 128.7 (CH), 131.5 (CH),
135.6 (2CH), 139.6 (C), 148.2 (C), 154.1 (C), 158.9 (C), 167.1
Sodium current was elicited by depolarizing pulses from a
holding potential value of -110 to -30 mV delivered at a
frequency of 0.2 Hz. To verify the stability of voltage clamp,
every five pulses, the holding potential was shifted to -90 mV
for one pulse. Sodium current was elicited by square depolariz-
ing pulses of 350 ms duration from a holding potential of -110
to -30 mV delivered at a frequency of 0.2 Hz. Sodium current
presenting an incomplete inactivation was induced with the
alkaloid veratridine (40 μM). The late INa was measured as the
mean current amplitude of the last 10 ms of the pulse (that is the
magnitude of INa at 340 to 350 ms of the depolarizing pulse).
Wild-type HEK293 cells did not exhibit any peak and late
inward sodium current. The compound was dissolved in 50%
DMSO in distilled water as 10 mmol stock solution prepared
freshly for each experiment. The final concentration of DMSO
was 0.1%. All values are expressed as means ( SEM.
Veratridine-Induced Contracture in Rats Isolated Atria. Male
Wistar rats (400-450 g; OFA, Iffa-Credo, France) were main-
tained at 20 ( 3 °C with constant humidity of 55 ( 5%, a 12 h
light-dark cycle, and free access to food and tap water. The
procedure is adapted from that described by Tamareille.53 After
a 30 min equilibration period, a single concentration of drug or
vehicle was injected into the organ bath. Fifteen minutes later,
veratridine (40 μM) was added. Systolic isometric tension
development was measured before drug or solvent injection
and just before the addition of veratridine in order to detect any
positive or negative inotropic effects of drug or vehicle. The
maximum amplitude of the veratridine-induced contracture was
determined irrespective of the time at which it occurred.
25
(2C); [R]D 0° (c 0.22, CH3OH). HPLC: eluting with acetoni-
trile/water/KH2PO4, 250:750:6.8 g, tR 33.9 min, 93.0%. Anal.
(C20H22N2S2O2 C4H4O4) C, H, N.
3
3,4-Dihydro-N-[(2S)-3-[indol-7-thio]-2-methylpropyl]-2H-
(3R)-1,5-benzoxathiepin-3-amine (2s). The free base was
purified by flash column chromatography (silica gel, di-
chloromethane/ethylacetate, 95:5). Crystallization of 2s as
a maleate salt gave a white powder mp=115 °C. IR (KBr) ν
1
3411, 1617, 1571, 1473, 1354 cm-1. H NMR (DMSO-d6) δ
1.10 (d, J = 6.8 Hz, 3H), 2.07 (m, 1H), 2.87-2.99 (m, 2H),
3.11-3.20 (m, 2H), 3.25 (d, J = 5.6 Hz, 2H), 3.77 (s, 1H),
4.34 (d, J = 13.2 Hz, 1H), 4.43 (d, J = 13.6 Hz, 1H), 6.04
(s, 2H), 6.50 (s, 1H), 6.97-7.08 (m, 3H), 7.22-7.26 (m, 2H),
7.35-7.40 (m, 2H), 7.50 (d, J = 7.6 Hz, 1H), 11.12 (s, 1H).
13C NMR (DMSO-d6) δ 17.2 (CH3), 30.9 (CH2), 30.9 (CH),
37.9 (CH2), 49.7 (CH2), 57.6 (CH), 70.4 (CH2), 101.9 (CH),
116.6 (C), 119.4 (CH), 119.6 (CH), 121.9 (CH), 124.1 (CH),
124.2 (CH), 125.8 (CH), 126.1 (C), 127.9 (C), 128.7 (CH),
131.4 (CH), 135.6 (2CH), 135.9 (C), 158.9 (C), 167.1 (2C);
Global Ischemia in Guinea Pig Isolated Hearts. Female guinea
pigs (SPF, Hartley, Charles River, France), weighing 500-
600 g, were used. The experiment was performed as described
previously.33 Briefly, hearts excised from guinea pigs were
placed in cold (4 °C) modified Krebs medium that contained
(in mmol): 124.6 NaCl, 4KCl, 1.1 MgSO4, 0.3 NaH2PO4, 1.8
CaCl2, 24.9 NaHCO3, and 11.1 D-glucose, pH 7.4, continuously
gases with 95% O2+5% CO2 and mounted on a Langendorff
system. A latex balloon was inserted through the left atrium and
mitral valve into the left ventricle. Isovolumetric systolic and
diastolic LVP, left ventricular developed pressure (LVDP =
systolic-diastolic pressure), left ventricular end diastolic pres-
25
[R]D 25.3° (c 0.42, CH3OH). HPLC: eluting with acetoni-
trile/water/KH2PO4, 400:600:6.8 g, tR 10.37 min. Anal.
(C21H24N2S2O C4H4O4) C, H, N.
3
3,4-Dihydro-N-[(2S)-3-[indazol-7-thio]-2-methylpropyl]-2H-
(3R)-1,5-benzoxathiepin-3-amine (2t). The free base was pur-
ified by flash column chromatography (silica gel, cyclohexane/
ethylacetate, 70:30). Crystallization of 2t as a maleate salt gave
a white powder mp = 127 °C. IR (KBr) ν 3059, 2959, 1610,
1
1572, 1473, 1355 cm-1. H NMR (DMSO-d6) δ 1.11 (d, J =
6.4 Hz, 3H), 2.11 (m, 1H), 2.98 (dd, J = 12.8, 7.6 Hz, 2H),
3.19-3.27 (m, 4H), 3.80 (s, 1H), 4.34 (d, J = 13.2 Hz, 1H), 4.42
(d, J = 12.8 Hz, 1H), 6.04 (s, 2H), 7.03-7.14 (m, 3H), 7.25 (t,
J = 5.6 Hz, 1H), 7.39 (d, J = 7.2 Hz, 1H), 7.44 (d, J = 6.8 Hz,
1H), 7.70 (d, J = 8.0 Hz, 1H), 8.16 (s, 1H), 13.5 (s, 1H). 13C
NMR (DMSO-d6) δ 17.1 (CH3), 30.9 (CH2), 31.1 (CH), 37.8
(CH2), 49.7 (CH2), 57.6 (CH), 70.4 (CH2), 118.7 (CH), 119.4
(CH), 120.9 (CH), 121.8 (CH), 122.3 (C), 123.1 (C), 124.1
(CH), 126.2 (C), 127.6 (CH), 128.7 (CH), 131.4 (CH), 134.1
sure, heart rate, positive dP/dt-1max, negative dP/dt-1
and
max
coronary flow were measured at 37 ( 1 °C with a constant
perfusion pressure of 800 mm H2O. The following parameters
were measured: systolic and diastolic left ventricular pressure,
left ventricular developed pressure, heart rate, positive dP/dtmax
,
negative dP/dtmax, and coronary flow. During perfusion under
constant pressure, changes in coronary flow directly reflect
changes in coronary vascular resistance. After 45-60 min
equilibration, the buffer solution containing vehicle or com-
pound was perfused for 15 min. Global normothermic ischemia
was then induced by clamping coronary flow for 50 min and was
followed by 60 min reperfusion. One drug concentration was
evaluated per heart.
25
(C), 135.6 (2CH), 158.9 (C), 167.1 (2C); [R]D -9.9° (c 0.41,
CH3OH). HPLC: eluting with acetonitrile/water/KH2PO4,
400:600:6.8 g, tR 5.25 min, 91.8%. Anal. (C20H23N3S2O
C4H4O4) C, H, N.
3
Biology. Animals used in this study were housed and tested in
an Association for the Assessment and Accreditation of La-
boratory Animal Care (AAALAC)-accredited facility in strict
compliance with all applicable regulations and the protocol was
carried out in compliance with French regulations and institu-
tional Ethical Committee guidelines for animal research. Car-
iporide51 and ivabradine52 were prepared according to literature
procedures; veratridine and lidocaine were purchased from
Sigma and TTX from Tocris Bioscience.
Ion Channel Pharmacology. The cell culture and patch clamp
experiments were performed as described previously.30 The
internal solution (pipet) contained (in mmol): NaCl 10, CsCl
110, CaCl2 1, HEPES 10, EGTA 10, Mg-ATP 5, D-glucose 10,
pH = 7.3 (CsOH). The external solution contained for patch
clamp experiments (in mmol): NaCl 30, CsCl 100, MgCl2 2,
CaCl2 2, HEPES 10, D-glucose 5, pH=7.4 (CsOH). All experi-
ments were carried out at room temperature (19-22 °C).
Myocardial Infarction Reperfusion in the Pig. Forty-five male
ꢁ
Landrace pigs (18-25 kg, GAEC La Jonione, Soreze, France)
were pretreated with an intramuscular injection of azaperone
(2 mg/kg) after an overnight fast. Anaesthesia was induced by
thiopental sodium (Nesdonal, 15 mg/kg) via the ear vein and
anesthesia was maintained with isoflurane (0.5-3% end-tidal
volume). After tracheal intubation with a cuffed endotracheal
tube, the lungs were ventilated with a positive pressure respira-
tor (alpha 100, Minerve, Esternay, France). Respiratory rate
and tidal volume were adjusted so as to maintain blood gases
within physiological limits (ABL 510, Radiometer, Copenhagen,
Denmark). Body temperature was kept constant between 37 and
38.5 °C by means of a homeothermic blanket. A four-limb ECG
was recorded in lead II throughout the experiment. Left lateral
thoracotomy was performed in the fourth intercostal space and
the pericardium opened. Polyethylene fluid-filled catheters were

4158 Journal of Medicinal Chemistry, 2009, Vol. 52, No. 14
Le Grand et al.
introduced into the nearest mammary artery to measure arterial
pressure during surgery and for blood sampling and into the
mammary vein to administer drugs or vehicle. All pigs were
treated with heparin (2500 IU) before the thorax was closed.
The proximal left circumflex (LCX) coronary artery was dissected
and a cotton ribbon placed around it. The LCX was occluded at
the prepared site for 60 min and then reperfused for 48 h. Thirty
minutes after the onset of reperfusion, the chest was closed in
layers and the pig was allowed to recover. Analgesia was achieved
with bupivacaine (2 mg/kg) and buprenorphine (0.05 mg/kg) at
regular intervals, or whenever signs of discomfort occurred.
Measurement of Infarct Size. After 48 h reperfusion, animals
were euthanized by sodium pentobarbital (160 mg/kg) and the
heart was excised. The right coronary, LCX, and left anterior
descending coronary arteries were catheterized. Evans blue
(0.4%) was infused into the right and left descending coronaries
to outline the ischemic myocardium. 2,3,5-Triphenyltetrazo-
lium chloride (0.1%) at 37 °C was infused into the LCX during
10 min so as to stain the viable myocardium red to measure the
extent of myocardial necrosis. The atria and the right ventricle
were removed. The left ventricle was then immersed in formal-
dehyde (10%) solution for at least 24 h. The left ventricle was cut
into 10 mm slices. Individual slices were photographed and the
extent of myocardial necrosis and the area at risk quantified by
image analysis software (Leica, Microsystems Imaging, Solu-
tions, Cambridge, United Kingdom).
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Measurements of Troponin I Plasma Levels. During reperfu-
sion, 10 blood samples were obtained from the mammary artery,
5 min before LCX artery occlusion, at the midpoint of LCX
artery occlusion (30 min), and at 5, 15, and 30 min and 1, 6, 24,
and 48 h reperfusion. The quantitative determination of tropo-
nin I in pig plasma was performed by AxSYM System (Abbot
Laboratories) in a medical analysis laboratory. The AxSYM
myoglobin and troponin I assays were based on microparticle
enzyme immunoassay technology. Intergroup statistical analyze
of results (P2, drug versus vehicle) was performed using one-way
analysis of variance followed by Dunnett’s test if ANOVA was
significant. Any P value lower than 0.05 was considered sig-
nificant (StatView 4.1, Abacus Concepts, Berkeley, CA).
Acknowledgment. We thank J. L. Maurel and S. Brunel,
who contributed to this project. We also thank Dr. J. P. Ribet
and P. Zalavari for analytical supports and L. Petitpas’
assistance for bibliographic searches.
Supporting Information Available: Intermediates 4a-f, 4h, 4i,
4l, 4o-q, 4u, 4v, 4t, 7, 8, 9j, 9m, and 9m, preparation of 2w, cell
culture and isolation, variations on basal hemodynamics in the
isolated perfused guinea pig heart, elemental analyses. This
material is available free of charge via the Internet at http://
pubs.acs.org.
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