New antiproliferative benzoindolinothiazepines derivatives

Article abstract of DOI:10.1016/j.ejmech.2004.10.008

New benzoindolinothiazepines containing a piperazine moiety are described as potent antiproliferative agents against PC3 human prostatic cell lines. This activity could be explained by an accumulation of cells in G1 phase.

Full text of DOI:10.1016/j.ejmech.2004.10.008

European Journal of Medicinal Chemistry 40 (2005) 167–172
www.elsevier.com/locate/ejmech
Original article
New antiproliferative benzoindolinothiazepines derivatives
Guillaume Laconde, Patrick Depreux *, Pascal Berthelot,
Nicole Pommery, Jean-Pierre Hénichart
Institut de chimie pharmaceutique Albert Lespagnol, universite de Lille 2, EA 2692, 3, rue du Professeur Laguesse, B.P. 83, 59006 Lille cedex, France
Received 8 April 2004; revised and accepted 11 October 2004
Available online 16 December 2004
Abstract
New benzoindolinothiazepines containing a piperazine moiety are described as potent antiproliferative agents against PC3 human prostatic
cell lines. This activity could be explained by an accumulation of cells in G1 phase.
© 2004 Elsevier SAS. All rights reserved.
Keywords: Benzoindolinothiazepines; Cell cycle; PC3
1. Introduction
during G1, is frequently mutated in human retinoblastoma
and lung cancer.
Deletion and mis-sense mutations result in truncated, non-
functional pRb or in complete absence of pRb. Approxi-
mately 90% of human cancers have abnormalities in some
components of the pRb pathway and p16/pRb alterations
occur commonly in primary and metastatic human prostate
cancer [12]. On the other hand, it has been found that some
FTase inhibitors (FTI) are able to induce apoptosis of trans-
formed cells. FTIs were designed to target Ras isoforms (H-,
N- and K-RasA/K-RasB) mutated in large number of cancers
and which require farnesylation for inducing uncontrolled cell
proliferation [13]. However, it is now demonstrated that Ras
is not the unique target of FTIs [14,15]. Several proteins could
be putative targets including Rho B which regulates receptor
trafficking and whose activation leads to p21waf1/Cip1 inacti-
vation and cell cycle progression to S phase [16] and centro-
meric proteins CENP-E and CENP-F [17]. Rho B exists in
both farnesylated and geranylgeranylated forms with gera-
nylgeranylated predominating Rho B, and FTI treatment
induces a loss of farnesylated Rho B and a gain of proapop-
totic geranylgeranylated Rho B [18]. FTIs also block the far-
nesylation of CENP-E and CENP-F, which function in the
mitotic spindle checkpoint of the cell cycle. Treatment of the
cells with FTIs decreases the association of CENP-E with
spindle microtubules and disrupts CENP-F localization [19].
In addition, FTI treatment results in failure of mitotic cells to
form bipolar spindles and to align chromosomes along the
metaphase plate [17]. These effects are likely to contribute to
Cell cycle control in G1 phase has attracted considerable
attention in recent cancer research, since key proteins involved
in G1 progression or G1/S transition have been found to play
a crucial role in proliferation, differentiation, transformation,
and apoptosis [1–3]. Particular attention has been focused on
cyclin-dependent kinases (CDKs) [4] and farnesyltrans-
ferase (FTase) [5]. The transition from one cell cycle phase to
another occurs in an orderly fashion and is regulated by regu-
latory proteins, the CDKs. These proteins, a family of
serine/threonine protein kinases, are activated at specific
points of the cell cycle [6]. CDK protein levels remain stable
during the cell cycle, in contrast to their activating proteins,
the cyclins. Cyclin protein levels rise and fall during the cell
cycle and in this way they periodically activate CDK [7].
Alterations of CDKs in cancer have been reported and the
process of searching for new cancer drugs is a direct inhibi-
tion of CDK [8]. The well known CDK inhibitors, olomou-
cine [9], roscovitine [10] and flavopiridol [11], all arrest pro-
liferation cells at the G1/S and G2/M boundaries and induce
apoptosis. Olomoucine also triggers an apoptotic response in
cells, which have been arrested in G2 phase by DNA-
damaging agents, whereas flavopiridol can induce apoptosis
in non-cycling cells. pRb, the most important CDK substrate
* Corresponding author. Tel.: +33 3 20 96 43 79; fax: +33 3 20 96 49 06.
E-mail address: pdepreux@phare.univ-lille2.fr (P. Depreux).
0223-5234/$ - see front matter © 2004 Elsevier SAS. All rights reserved.
doi:10.1016/j.ejmech.2004.10.008

168
G. Laconde et al. / European Journal of Medicinal Chemistry 40 (2005) 167–172
Fig. 1. Structure of SCH-44342, E7070 and hybrid scaffold.
the G2/M arrest observed in FTI treated cells and may con-
tribute to the synergistic antitumor interaction between FTIs
and taxanes, anticancer drugs that also target the mitotic appa-
ratus [20]. Other FTI effects on farnesylation of nuclear enve-
lope proteins lamin A and B, nuclear tyrosine phosphatases
(PTPCAAX 1, 2 and 3) and cochaperone protein HDJ-2, are
likely involved in their antitumor activity [21]. On the other
hand, in a recent study, it has been claimed that CDK inhibi-
tors dramatically enhance FTI’s induced apoptosis of human
cancer cell lines. Thus, it was tempting to design new hybrid
molecules able to possess these two sets of properties [22].
Considering SCH-44342, a potent inhibitor of FTase [23]
and E7070 [24], a suppressor of the expression of cyclin E
and inhibitor of CDK2 and pRb phosphorylations (Fig. 1),
we designed tetracyclic new compounds combining struc-
tural features of 1 and 2. Here are reported the synthesis, the
antiproliferative effect and the G1 phase study of these com-
pounds against PC3 tumor cell lines.
2. Chemistry
1-Acetyl-5-chlorobenzo[f]indolino[6,5,c] [1,2]thiazepine
(12) was prepared as described in Fig. 2. The nitration of 3
was achieved by using a mixture of nitric acid and sulfuric
acid (yield 80%) yielding 4 which subsequently formed
1-acetyl-6-nitroindoline by reaction with acetic anhydride
(yield 95%) [25]. The reduction of nitro entity in amino was
realized by hydrogenation with palladium on activated car-
bon (yield 95%). The sulfonamide compound was obtained
by coupling 6 with methyl 2-chlorosulfonylbenzoate in pyri-
dine (yield 72%) and exposure of 7 to methyl iodide gave
Nmethylsulfonamide (yield 70%). Treatment of 8 by potas-
sium hydroxide gave the carboxylic acid (yield 90%) and the
cyclization of 9 under Friedel–Crafts conditions with alumi-
num chloride resulted in the formation of the expected tetra-
cycle 10 (yield 41%).
Fig. 2
. (a) HNO3, H2SO4, r.t.; (b) Ac2O, 160 °C; (c) H2, Pd/C 10%, MeOH, r.t.; (d) methyl 2-chlorosulfonylbenzoate, pyridine, DMF, 65 °C; (e) CH3I, NaH,
DMF, r.t.; (f) KOH, H2O/EtOH, 100 °C; (g) (1) SOCl2, toluene, 80 °C (2) AlCl3, CHCl3, 60 °C; (h) NaBH4, MeOH/CH2Cl2, 50 °C; (i) SOCl2, CH2Cl2, r.t.

G. Laconde et al. / European Journal of Medicinal Chemistry 40 (2005) 167–172
169
NMR studies confirmed the structure of these new tetra-
3. Pharmacological studies
cyclic system. The ketone 10 was reduced by sodium boro-
hydride (yield 65%), then the hydroxylated compound 11 was
treated by thionyl chloride to form 12 (yield 74%).
3.1. Cell culture and cytotoxicity
Human prostate cancer PC3 cells were maintained in RPMI
1640 culture medium supplemented with 10% FCS. For
growth assays, the cells were seeded onto 96-well plates at a
density of approximately 3 × 104 cells per well. After 3 days,
the cell medium was changed to serum-free medium and the
cells were starved for 24 h for culture synchronization. The
stimulation of the growth of quiescent cells was then per-
formed by 10 ng/ml EGF plus TSe (50 pg/ml selenium) and
the tested compounds were added to culture medium. After
an additional 72 h, the cell growth was estimated by the colo-
rimetric MTT test (Table 1).
The piperazine derivatives have been prepared by two dif-
ferent methods (Fig. 3). Method A: The Nbenzylpiperazine
was coupled with commercially acyl chloride using triethy-
lamine to afford N-acylpiperazine derivative in good yield.
The deprotection was realized by hydrogenation with palla-
dium on activated carbon. Method B: The N-benzyl-
piperazine was coupled with commercially acid available
using NMM, EDCI, HOBt to afford N-acylpiperazine deriva-
tive in good yield. The deprotection was achieved by hydro-
genation with palladium on activated carbon.
The synthesis of compounds 16a–f was carried out. The
final substitution of C–Cl was achieved by reaction of five
equivalents of appropriate amine 14 and 15 with 12 in aceto-
nitrile at 110 °C for 1 h (Fig. 4).
3.2. Cell cycle and apoptosis
The PC3 cells were grown at a density of about 5 × 105
cells on 25 cm2 dishes, synchronized during 24 h in serum-
free medium and stimulated by EGT–TSe in the presence of
different tested agents for 3 days. An analysis by FACS was
then performed using the method described in the cell cycle
test kit from Becton Dickinson. The propidium iodide-
stained cell populations in sub-G1 (corresponding to sub-
diploid nuclei), G1, S and G2/M phases were quantified using
the Cellquest logiciel program (Table 2).
4. Results and discussion
The aim of this work was to gather on the same scaffold
by pharmacophoric elements of reference compounds of
SCH-44342 and E7070 in view of designing new heterocy-
clic compounds able to exhibit interesting cytostatic proper-
ties. Two objectives were fixed to qualify these new mol-
ecules: (i) they had to present relevant IC50 in terms of
proliferation inhibition and (ii) their mechanism of action
would imply an arrest of cell cycle. The first point is reached
Fig. 3. Method A (a) acyl chloride, Et3N, CH2Cl2, r.t.; (b) H2, Pd/C 10%,
MeOH, r.t.; Method B (c) pyridacetique hydrochloric acid, NMM, HOBt,
EDCI, DMF, r.t. (d) H2, Pd/C 10%, MeOH, r.t.
Table 1
Activity of compounds 16a–16f (1 µM) on PC3 cell proliferation (% inhibi-
tion)
Compounds
16a
R
PC3
CH3
29.6%
16b
16c
16d
16e
CH3CO
IC50 = 0.19 µM
IC50 = 6.49 µM
IC50 = 9.85 µM
43.10%
CH3CH2CO
CH3CH2CH2CO
1-(pyridin-4-yl-acetyl)
1-(pyridin-3-yl-acetyl)
16f
37.94%
Table 2
Effects of 16b on treated PC3 cell cycle
Compounds
Cell populations in cycle phases (%)
Sub-G1
5.41
G1
S
G2/M
19.69
15.93
Control
65.30
74.68
9.86
4.68
16b
5.27
Fig. 4. (a) CH3CN, 110 °C.

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G. Laconde et al. / European Journal of Medicinal Chemistry 40 (2005) 167–172
since one of the benzoindolinothiazepines (16b) inhibits
PC3 proliferation with an IC50 of 0.19 µM. It is known that
PC3 cells responsible of invasive aspects of prostate cancer,
are highly resistant to classical therapies and these primary
results are very promising. Moreover, a set of structure–
activity relationship could be delineated: an alkyl group on
piperazinyl ring seems necessary for a reasonable prolifera-
tion inhibition and aromatic substituents are not favorable. In
addition, it is clear that an amide group on distal piperazinyl
N-atom increases the antiproliferative properties. The sec-
ond point deals with the ability of these compounds to induce
accumulation of PC3 cells in G1 phase. Such is the case for
16b and this result is in accordance with our hypothesis. This
hybrid molecule behaves like E7070 and it may be that it also
acts on targets of certain FTI since it has been reported that
SCH-663365 (SARAMAR), an analog of SCH-44342, is an
inhibitor of CENP-E and CENP-F prenylation in A549 lung
carcinoma cells lines altering association between CENP-E
and microtubules and affecting microtubulecentromere inter-
action and also is an inhibitor of FTase in H-ras transformed
cells to accumulation in G1 phase [20]. The concept of hybrid
molecules developed here is also in accordance with the
principe of combination of FTI with standard and mitotic
agents such as FTI/docetaxel in the treatment of breast can-
cer and of FTI/paclitaxel in the treatment of prostate cancer
[26,27].
under reduced pressure. The residue was taken up in water,
stirred at r.t. for 1 h and the precipitate formed was filtered.
The product was purified by recrystallization from methanol.
Yield 72%. 7: As white crystals, m.p. 165–166 °C (metha-
nol). IR (KBr): mmax = 3230 (NH) and 1350, 1160 (SO2N)
cm–1; 1H NMR (300 MHz, CDCl3, Me4Si): d = 2.16 (3H, s,
CH3CO), 3.11 (2H, t, J = 8.36 Hz, CH2), 4.02 (2H, t,
J = 8.36 Hz, CH2), 4.04 (3H, s, COOCH3), 7.05 (2H, m,ArH),
7.55 (2H, m, ArH), 7.86 (3H, m, ArH), 8.10 (1H, m, NH);
MS: m/z (%) = 374 (100).
5.3. Methyl N-(1-acetylindolin-6-yl)-N-methyl-2-sulfamoyl
benzoate (8)
Methyl N-(1-acetylindolin-6-yl)-2-sulfamoyl benzoate (7)
(7.40 g, 0.019 mol) in DMF was added dropwise to a solu-
tion of NaH (1.52 g, 0.038 mol) in DMF. The solution was
stirred at r.t. while a red color slowly developed. After 3 h,
CH3I diluted in DMF was added dropwise over 10 min and
the mixture was stirred for 16 h at r.t. The solution was evapo-
rated under reduced pressure. The residue was taken up in
water, stirred at r.t. for 1 h and the precipitate formed was
filtered. The product was purified by recrystallization from
methanol. Yield 70%. 8: As white crystals, m.p. 163–164 °C
1
(methanol); IR (KBr): mmax = 1350, 1170 (SO2N) cm–1; H
NMR (300 MHz, CDCl3, Me4Si): d = 2.20 (3H, s, CH3CO),
3.19 (2H, t, J = 8.47 Hz, CH2), 3.28 (3H, s, NCH3), 3.90 (3H,
s, COOCH3), 4.11 (2H, t, J = 8.47 Hz, CH2), 7.05 (1H, m,
ArH), 7.15 (1H, d, J = 7.92 Hz, ArH), 7.55 (4H, m, ArH),
7.94 (1H, m, ArH); MS: m/z (%) = 388 (100).
5. Experimental protocols
Melting points (m.p.) were determined on a Büchi SMP-
540 apparatus. 1H NMR spectra were recorded on anAC 300 P
(300 MHz) spectrometer using d6-DMSO or CDCl3 as sol-
vents. Chemical shifts are expressed downfield from the inter-
nal standard, tetramethylsilane. Coupling constants (J) are
expressed in Hz. Key: t = triplet, s = singlet, d = doublet, dd
= double doublet, m = multiplet. Mass spectra were recorded
on a Funnigan Mat SSQ710 mass spectrometer. Elemental
analyses (C, H, N) were determined by the CNRS Center of
Analysis,Vernaison, France, and agreed with proposed struc-
tures within 0.4% of the theoretical values.
5.4. N-(1-acetylindolin-6-yl)-N-methyl-2-sulfamoyl benzoic
acid (9)
To Methyl N-(1-acetylindolin-6-yl)-N-methyl-2-sulfa-
moyl benzoate (8) (5.00 g, 0.013 mol) in ethanol/water (1:1)
was added potassium hydroxide (2.16 g, 0.038 mol). The
resulting mixture was stirred at 100 °C for 2 h. After cooling,
the reaction medium was added to water and acidified with
HCl 12 N for pH 1. The precipitate formed was filtered off
and recrystallized from MeOH.Yield 90%. 9: As white crys-
tals, m.p. 228–229 °C (methanol); IR (KBr): mmax = 3420 (OH)
and 1350, 1160 (SO2N) cm–1; 1H NMR (300 MHz, DMSO,
Me4Si): d = 2.12 (3H, s, CH3CO), 3.13 (2H, t, J = 8.49 Hz,
CH2), 3.15 (3H, s, NCH3), 4.11 (2H, t, J = 8.49 Hz, CH2),
6.77 (1H, m, ArH), 7.18 (1H, m, ArH), 7.38 (1H, m, ArH),
7.57 (2H, m, ArH), 7.69 (1H, m, ArH), 7.92 (1H, m, ArH),
13.50 (1H, m, COOH); MS: m/z (%) = 374 (100).
5.1. N-acetyl-6-nitroindoline (5) and N-acetyl-6-
aminoindoline (6)
Compounds 5 and 6 were prepared according to a proce-
dure already described [25].
5.2. Methyl N-(1-acetylindolin-6-yl)-2-sulfamoyl benzoate
(7)
5.5. 1-Acetyl-5,11-dihydro-11-methylbenzo[f]indolino[6,5,c]
[1,2]thiazepin-5-one 10,10-dioxyde (10)
Methyl 2-chlorosulfonylbenzoate (7.62 g, 0.033 mol) in
DMF was added dropwise to a solution of 1-acetyl-6 ami-
noindoline (6) (6.88 g, 0.039 mol) in DMF and pyridine
(3.15 g, 0.039 mol) at room temperature (r.t.). The reaction
mixture was stirred at 65 °C for 60 min and then evaporated
To N-(1-acetylindolin-6-yl)-N-methyl-2-sulfamoyl ben-
zoic acid (9) (2.00 g, 0.005 mol) in toluene was added thionyl
chloride (1.11 ml, 0.016 mol). The reaction was refluxed for
1 h.After cooling, the reaction medium was evaporated under

G. Laconde et al. / European Journal of Medicinal Chemistry 40 (2005) 167–172
171
reduced pressure. The resulting mixture was taken up in chlo-
5.9. Method B: General procedure for the preparation
of 1-acylpiperazines (15e–15f)
roform. Aluminum chloride was added (2.12 g, 0.016 mol)
and refluxed for 1 h. After cooling, the reaction was evapo-
rated under reduced pressure. The resulting mixture was taken
up in water and extracted with dichloromethane. The organic
layer was washed with NaOH 1 N, water and dried with mag-
nesium sulfate. The precipitate was recrystallized from EtOH.
Yield 41%. 10: m.p. 228–229 °C (ethanol); IR (KBr): mmax
= 1650 (CO) and 1320, 1170 (SO2N) cm–1; 1H NMR
(300 MHz, CDCl3, Me4Si): d = 2.31 (3H, s, CH3CO), 3.28
(2H, t, J = 8.45 Hz, CH2), 3.31 (3H, s, NCH3), 4.17 (2H, t,
J = 8.45 Hz, CH2), 7.72 (2H, m, ArH), 7.97 (2H, m, ArH),
8.15 (1H, s, ArH), 8.21 (1H, s, ArH); MS: m/z (%) = 356.
To N-benzylpiperazine (13) (0.50 g, 0.002 mol) in dime-
thylformamide was added N-methylmorpholine (1.87 g,
0.017 mol), 1-hydroxybenzotriazole (0.38 g, 0.002 mol), 1-(3-
dimethylaminopropyl)-3-ethylcarbo diimide (0.81 g,
0.004 mol) and pyridylacetic acid hydrochloride (0.56 g,
0.003 mol). The reaction was stirred for 2 days and was evapo-
rated under reduced pressure. The resulting mixture was taken
up in dichloromethane and washed with potassium hydrogen-
carbonate solution and water. The organic layer was dried
with magnesium sulfate. The white oil was used for further
purification in the next step.
5.6. 1-Acetyl-5,11-dihydro-11-methylbenzo[f]indolino
[6,5,c] [1,2]thiazepin-5-ol 10,10-dioxyde (11)
5.10. General procedure for the preparation of 1-acetyl5-
(1-acylpiperazine)-5,11-dihydro-11-
To 1-acetyl-5,11-dihydro-11-methylbenzo[f]indolino
[6,5,c] [1,2] thiazepin-5-one 10,10-dioxyde (10) (0.80 g,
0.002 mol) in dichloromethane was added sodium borohy-
dride (0.13 g, 0.003 mol). The reaction was refluxed for 2 h.
After cooling, water was added dropwise to the reaction and
the resulting mixture was filtered. Yield 65%. 11: m.p. 189–
190 °C; IR (KBr): mmax = 1650 (CO) and 1320, 1160 (SO2N)
cm–1; 1H NMR (300 MHz, CDCl3, Me4Si): d = 2.13 (3H, s,
CH3CO), 3.13 (2H, t, J = 8.45 Hz, CH2), 3.37 (3H, s, NCH3),
4.10 (2H, t, J = 8.45 Hz, CH2), 6.20 (1H, s, ArH), 6.51 (1H,
m, OH), 7.41 (1H, s, ArH), 7.48 (1H, m, ArH), 7.61 (1H, m,
ArH), 7.80 (2H, m, ArH), 8.15 (1H, s, ArH); MS: m/z (%)
= 358.
methylbenzo[f]indolino[6,5,c] [1,2]thiazepine 10,10-
dioxyde (16a–16f)
To 1-Acetyl-5-chloro-5,11-dihydro-11-methylbenzo[f]
indolino[6,5,c]
[1,2]thiazepine
10,10-dioxyde
(12)
(0.0002 mol) in acetonitrile was added l-acylpiperazine
(0.001 mol). The reaction was stirred for 1 h and was evapo-
rated under reduced pressure. The resulting mixture was taken
up in water and extracted with dichlromethane. The organic
layer was washed with NaOH 1 N and dried with magnesium
sulfate. The precipitate was recrystallized from EtOH.
5.10.1. 1-Acetyl-5-(1-methylpiperazine)-5,11-dihydro-11-
methylbenzo[f]indolino[6,5,c] [1,2]thiazepine 10,10-
dioxyde (16a)
5.7. 1-Acetyl-5-chloro-5,11-dihydro-11-methylbenzo[f]-
indolino[6,5,c] [1,2]thiazepine 10,10-dioxyde (12)
Yield 55%. M.p. 154–155 °C; 1H NMR (300 MHz), 2.00
(3H, s), 2.20 (7H, m), 3.25 (5H, m), 3.55 (3H, s), 3.69 (1H,
m), 4.08 (3H, m), 7.08 (1H, s), 7.30 (1H, dd), 7.44 (2H, m),
7.99 (1H, dd), 8.32 (1H, s); IR (KBr): 2980, 1670, 1580,
1340 cm–1; Anal. (C23H28N4O3S).
1-Acetyl-5,11-dihydro-11-methylbenzo[f]indolino[6,5,c]
[1,2]thiazepin-5-ol 10,10-dioxyde (11) (0.41 g, 0.001 mol) in
dichloromethane was added thionyl chloride (0.12 ml,
0.002 mol). The reaction was stirred overnight. The resulting
mixture was filtered. Yield 74%. 12: m.p. 112–113 °C; IR
(KBr): mmax = 1670 (CO) and 1350, 1160 (SO2N) cm–1; 1H
NMR (300 MHz, CDCl3, Me4Si): d = 2.16 (3H, s, CH3CO),
3.10 (2H, t, J = 8.45 Hz, CH2), 3.34 (3H, s, NCH3), 4.10 (2H,
t, J = 8.45 Hz, CH2), 6.20 (1H, s, ArH), 7.41 (1H, s, ArH),
7.48 (1H, m, ArH), 7.62 (1H, m, ArH), 7.80 (2H, m, ArH),
8.10 (1H, s, ArH); MS: m/z (%) = 376.
5.10.2. 1-Acetyl-5-(1-acetylpiperazine)-5,11-dihydro-11-
methylbenzo[f]indolino[6,5,c] [1,2]thiazepine 10,10-
dioxyde (16b)
Yield 70%. M.p. 163–164 °C; 1H NMR (300 MHz), 2.00
(3H, s), 2.20 (7H, m), 3.25 (5H, m), 3.55 (3H, s), 3.69 (1H,
m), 4.08 (3H, m), 7.08 (1H, s), 7.30 (1H, dd), 7.44 (2H, m),
7.99 (1H, dd), 8.32 (1H, s); IR (KBr): 2940, 1680, 1580,
1330 cm–1; Anal. (C24H28N4O4S).
5.8. Method A: General procedure for the preparation
of 1-acylpiperazines (14b–14d)
5.10.3. 1-Acetyl-5-(1-propionylpiperazine)-5,11-dihydro11-
methylbenzo[f]indolino[6,5,c] [1,2]thiazepine 10,10-
dioxyde (16c)
To N-benzylpiperazine (13) (2.00 g, 0.011 mol) in dichlo-
romethane was added triethylamine (0.95 ml, 0.011 mol) and
dropwise acylchloride (0.017 mol). The reaction was strirred
overnight and was evaporated under reduced pressure. The
resulting mixture was taken up in water and extracted with
ethyl ether. The organic layer was dried with magnesium sul-
fate.
Yield 71%. M.p. 182–183 °C; 1H NMR (300 MHz), 1.12
(3H, m), 2.26 (9H, s), 3.25 (5H, m), 3.52 (3H, s), 3.70 (1H,
m), 4.09 (3H, m), 7.07 (1H, s), 7.29 (1H, m), 7.46 (2H, m),
7.99 (1H, dd), 8.32 (1H, s); IR (KBr): 2950, 1680, 1580,
1330 cm–1; Anal. (C25H30N4O4S).

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methylbenzo[f]indolino[6,5,c] [1,2]thiazepine 10,10-
dioxyde (16d)
Yield 68%. M.p. 178–179 °C; 1H NMR (300 MHz), 0.96
(3H, m), 1.63 (2H, m), 2.23 (9H, m), 3.25 (5H, m), 3.54 (3H,
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5.10.5. 1-Acetyl-5-(1-(pyridin-4-yl-acetyl)piperazine)5,11-
dihydro-11-methylbenzo[f]indolino[6,5,c] [1,2]thiazepine
10,10-dioxyde (16e)
Yield 55%. M.p. 182–183 °C; 1H NMR (300 MHz), 2.20
(7H, m), 3.12 (3H, m), 3.40 (6H, m), 3.70 (3H, s), 4.13 (2H,
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5.10.6. 1-Acetyl-5-(1-(pyridin-3-yl-acetyl)piperazine)5,11-
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10,10-dioxyde (16f)
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