Synthesis and anti-tumor activities of novel methylthio-, sulfinyl-, and sulfonyl-8H-thieno[2,3-b]pyrrolizin-8-oximino derivatives

Article abstract of DOI:10.1002/ardp.200700044

A series of novel methylthio-, sulfinyl-, and sulfonyl-8H-thieno[2,3-b] pyrrolizin-8-oximino derivatives 7A-12P was designed and synthesized as anti-tumor agents. Their structures were confirmed by IR, ¹H-NMR, MS, and elemental analysis. The anti-tumor activities of all the target compounds were tested by the MTT method in vitro against Bel-7402 (human liver cancer) and HT-1080 (human fibro sarcoma) cell lines. Among them, compound UN (IC ₅₀ = 18.2 μM, 8.2 μM), was the most promising compound of all synthesized molecules, it was 2.5- and 3.3-times more active than cisplatin (IC₅₀ = 45.2 μM, 26.7 μM), respectively.

Full text of DOI:10.1002/ardp.200700044

416
Arch. Pharm. Chem. Life Sci. 2007, 340, 416–423
Full Paper
Synthesis and Anti-tumor Activities of Novel Methylthio-,
Sulfinyl-, and Sulfonyl-8H-thieno[2,3-b]pyrrolizin-8-oximino
Derivatives
Shuchun Guo, Yanfang Zhao, Xianglin Zhao, Sisi Zhang, Lijun Xie, Weiyong Kong, and
Ping Gong
School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang, P. R. China
A series of novel methylthio-, sulfinyl-, and sulfonyl-8H-thieno[2,3-b]pyrrolizin-8-oximino deriv-
atives 7A–12P was designed and synthesized as anti-tumor agents. Their structures were con-
firmed by IR, 1H-NMR, MS, and elemental analysis. The anti-tumor activities of all the target com-
pounds were tested by the MTT method in vitro against Bel-7402 (human liver cancer) and HT-
1080 (human fibro sarcoma) cell lines. Among them, compound 11N (IC50 = 18.2 lM, 8.2 lM),
was the most promising compound of all synthesized molecules, it was 2.5- and 3.3-times more
active than cisplatin (IC50 = 45.2 lM, 26.7 lM), respectively.
Keywords: Anti-tumor activities / Synthesis / Thieno[2,3-b]pyrrolizines /
Received: March 6, 2007; accepted: June 1, 2007
DOI 10.1002/ardp.200700044
atives 1 acting as anti-tubulin inhibitors were found to
Introduction
possess good cell-growth inhibitory activity over a large
panel of human tumoral cell lines, including the resist-
ant KB-A1 cell line [11]. These features make the
thieno[2,3-b]pyrrolizines potential leads for new anti-
tumor agents. And the structure-activity relationship
(SAR) of 1 indicated that the attachment of a phenyl-bear-
ing, small substituent in the meta and para position to
position-3 on the tripentacyclic skeleton was essential for
cytotoxicity.
Cancer is a growing public problem estimated to have
worldwide about seven million new incidences per year.
Research oriented towards the discovery of a new genera-
tion of agents useful in anticancer chemotherapy has
resulted in the identification of various more potent
compounds. Among them, sulfur-containing com-
pounds, such as thiazole, thiophene, thioether, sulfona-
mide, sulfones, and thio-heterocyclic derivatives demon-
strated excellent anti-tumor activities. They have been
shown to function by different mechanisms of action,
including cyclin-dependent kinase inhibition [1], DNA
photocleavage [2–4], inhibition of Bcl-2 protein [5], and
inhibition of tubulin polymerization [6]. It is noted that
thieno[2,3-b]pyrrolizines, a new tripentacyclic skeleton,
attract increasing interest due to their diverse pharmaco-
logical effects, for instance, as CDK/GSK-3 inhibitors [7], 5-
HT3 and/or 5-HT2C agonists [8, 9], and anti-tumor agents
[10]. Recently, 3-arylthieno[2,3-b]pyrrolizin-8-one deriv-
With the aim of searching for new anti-tumor agents
and understanding their SARs, we designed and synthe-
sized a novel series of 2-alkylthio-, sulfinyl- and sulfonyl-
3-aryl-8H-thieno[2,3-b]pyrrolizin-8-oximino
derivatives
(Fig. 1), focusing our attention on modifying position-2
and position-8 on the thieno[2,3-b]pyrrolizine scaffold.
Alkylthio/sulfinyl/sulfonyl groups, which proved to be
effective moieties for the potent inhibition of tubulin
polymerization [6, 12], were introduced at position-2.
While various hydrophilic aminoalkoxyimino branching
chains, which were then replaced with aminoalkoxyme-
thyloxyimino substituents for further exploring the SAR,
were introduced at position-8. In light of the SAR men-
tioned above, a 4-nitrophenyl group was attached to posi-
tion-3 for preliminary study. Anti-tumor activities of all
Correspondence: Ping Gong, 103 Wenhua Road, Shenhe District,
110016 Shenyang, Liaoning, P. R. China.
E-mail: gongpinggp@126.com
Fax/Phone: +86 24 2388-2925
i 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Arch. Pharm. Chem. Life Sci. 2007, 340, 416–423
Novel Thieno[2,3-b]pyrrolizines
417
acid, according to the Clauson–Kaas method [14],
afforded the pyrrolylthiophene carboxylate 3, which was
converted into the corresponding carboxamide 4 in pyr-
rolidine at 708C. Cyclization of 4 in phosphorus oxychlor-
ide yielded an iminium chloride intermediate, which
was treated with an aqueous solution of sodium hydrox-
ide to yield the tricyclic ketone 5. Reaction of the latter
compound with hydroxylamine hydrochloride in pyri-
dine furnished the oxime 6 generally in a mixture of Z-
and E-forms without separation. By treatment of 6 with
appropriate aminoalkyl chloride prepared as published
[15, 16] and potassium carbonate, the oxime derivatives
7A-F were obtained.
Figure 1. Structure of thieno[2,3-b]pyrrolizin-8-oximino deriv-
atives 7A-12P.
synthesized compounds were evaluated by the MTT [3-
(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bro-
mide] method in vitro.
To further investigate the effects of the substituents at
position-8 of the tricyclic core, we introduced the amino-
alkoxymethyloxyimino groups by a two-step sequence
involving the addition of the oxime 6 to the 1-chloro-2-
(chloromethoxy)ethane in the presence of sodium metha-
nolate to give 8-(2-chloroethyloxymethyloxyimino)-2-
methylthio-3-(4-nitrophenyl)-8H-thieno[2,3-b]pyrrolizine
10 and then reaction with various secondary amines to
afford the desired compounds 11K-O.
Results and discussion
Chemistry
The title compounds methylthio-, sulfinyl- and sulfonyl-
8H-thieno[2,3-b]pyrrolizin-8-oximino derivatives were
obtained as described in Scheme 1. The substituents of
compounds 7A-12P are listed in Table 1.
The sulfur derivatives 7 and 11 were oxidized into the
corresponding sulfoxides 8G-H and 12P by one equiva-
lent of sodium perborate in acetic acid at room tempera-
ture. However, the sulfone derivatives were difficult to
accomplish here with the above oxidant as well as
MCPBA (m-chloroperoxybenzoic acid), because the nitro-
gen at position-8 would also be oxidized to form an unde-
The preparation of thiophene intermediate 2 was per-
formed in a one-pot reaction by treatment of (4-nitrophe-
nyl)acetonitrile with carbon disulfide in the presence of
sodium hydroxide in THF, via an intermediate iminium
salt, followed by successively reacting with methyl 2-
chloroacetate and methyl iodide in 53% yields [13]. Treat-
ment of 2 with 2,5-dimethoxytetrahydrofuran in acetic
Reagents and conditions: a) (i) NaOH / THF, r.t., (ii) ClCH2COOCH3/CH3I, – 208C; b) dimethoxy THF/CH3COOH, 708C; c) pyrrolidine, 708C; d) (i) POCl3, 808C, (ii) 10% NaOH,
608C; e) NH2OH6HCl, pyridine, 908C; f) ClCH2CH2NR1R26HCl/K2CO3, 708C; g) NaBO364 H2O/CH3COOH, 25l308C; h) H2O2/Na2WO462 H2O/CH3OH, 208C; i)
ClCH2OCH2CH2Cl/CH3ONa, 258C; j) HNR1R2/NaOH/DMF, 608C; k) NaBO364 H2O/CH3COOH, 25l308C.
Scheme 1. Synthesis of target compounds 7A-12P.
i 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

418
S. Guo et al.
Arch. Pharm. Chem. Life Sci. 2007, 340, 416–423
Table 1. The substituents of compounds 7A-12P.
Table 2. The anti-tumor activities of compounds 5-12P.
Compound
X
Y
NR1R2
Compound
IC50 (lM)
HT-1080
7A
7B
7C
7D
7E
7F
8G
8H
9I
-CH2-
-CH2-
-CH2-
-CH2-
-CH2-
-CH2-
-CH2-
-CH2-
-S-
-S-
-S-
-S-
-S-
-S-
-SO-
-SO-
-SO2-
-SO2-
-S-
-S-
-S-
-S-
-S-
morpholinyl
diethylamino
pyrrolidinyl
dimethylamino
piperdinyl
4-methylpiperazinyl
morpholinyl
diethylamino
morpholinyl
diethylamino
pyrrolidinyl
dimethylamino
piperdinyl
4-methylpiperazinyl
morpholinyl
morpholinyl
Bel-7402
5
6
7A
7B
7C
7D
7E
7F
8G
8H
9I
9J
11K
11L
11M
11N
11O
12P
Cisplatin
195.9
177.8
124.7
31.1
23.1
37.7
193.1
184.8
198.1
31.1
18.4
21.2
28.6
34.9
289.3
48.0
276.5
316.3
24.4
25.5
68.0
8.3
-CH2-
-CH2-
59.1
31.4
9J
11K
11L
11M
11N
11O
12P
-CH2OCH2-
-CH2OCH2-
-CH2OCH2-
-CH2OCH2-
-CH2OCH2-
-CH2OCH2-
178.9
55.5
263.3
298.4
69.6
26.4
27.2
18.2
98.3
-SO-
sired N-oxidized side-product. After an extensive survey
of the different reagents available, we have observed that
H2O2/Na2WO4 is very effective for oxidizing sulfide into
sulfone to furnish the compounds 9I-J in excellent yield
and high purity.
61.8
89.3
26.7
145.8
45.2
Among the compounds 7A-7F, 7C displayed the most
potent anti-tumor activity against the two cancer cell
lines. Nevertheless, 7A, which possessed the morpholino
substituent at the amino alkoxy moiety of position-8, was
6- or 10-times less active than 7C. Other molecules 7B, 7D-
F also exhibited comparable efficacy to the positive con-
trol cisplatin. It seemed that this region could tolerate a
hindered secondary amine, but the morpholino group
abolished the anti-tumor activity. In another set of 11K-
O, 8-[2-(4-methylpiperazinyl)ethyloxymethyloxyimino]-2-
methylthio-3-(4-nitrophenyl)-8H-thieno[2,3-b]pyrrolizine
11N showed IC50 = 18.2 lM, 8.2 lM, 2.5- and 3.3-times
more active than cisplatin (IC50 = 45.2 lM, 26.7 lM),
respectively and it was the most promising compound of
all the synthesized analogues. Except for 11K, the anti-
tumor activity against the Bel-7402 cell line was
improved by inserting methylene oxy group into the
amino alkoxy moiety (compare 11L with 7D, 11M with
7E, 11N with 7F, 11O with 7A). These might suggest that
expansion of the amino alkoxy moiety with a heteroatom
or carbon and introduction of suitable cyclic secondary
amines, especially 4-methyl piperazine, could enhance
the anti-tumor activity.
The Z/E-isomerism of oxime 6 and oxime derivatives
7A-12P were elucidated by H-NMR spectroscopy [8, 9].
1
The most pronounced changes were observed in the
chemical shifts of the H-7 and SCH3-2 protons of the thie-
nopyrrolizine ring in the Z- and E-isomers. (Z)-Oxime 6
showed a signal at 6.55 ppm, for H-7. In contrast, (E)-
oxime 6 exhibited the H-7 signal at a lower field,
6.80 ppm, owing to the deshielding effect of the hydroxyl
group [17] on the H-7. On the other hand, the chemical
shift of the SCH3-2 of (Z)-6 is influenced by the hydroxyl
group and the signal appeared downfield at 2.48 ppm,
whereas that of the (E)-6 showed the signal at 2.43 ppm.
Similar results were observed in the 1H-NMR of the oxime
derivatives 7A-12P. The H-7 proton of Z-form showed
around 6.53-6.64 ppm, whereas that of the E-form around
6.69–6.82 ppm, in good agreement with those of oxime
6. The Z/E-isomeric ratios were estimated from the inte-
gration of the resonance signals.
Assay for anti-tumor activity
Compounds 5-12P were evaluated for anti-tumor activ-
ities in vitro by the MTT method against Bel-7402 (human
liver cancer) and HT-1080 (human fibro sarcoma) cell
lines using cisplatin as positive control, and the results
expressed as IC50 were summarized in Table 2. As shown,
most of the evaluated compounds exhibited good anti-
tumor activities, and some were superior to cisplatin.
The O-substituted oximino-pyrrolizines exhibited more
potent anti-tumor activities than the corresponding tri-
pentone 5 and the oxime 6 in most cases.
Oxidation of the sulfur atom to the sulfoxide or the sul-
fone was found to be detrimental to the anti-tumor activ-
ity (compounds 7A vs 8G; 7B vs 8H; 11O vs 12P and 8G vs
9I; 8H vs 9J).
In conclusion, a series of novel methylthio-, sulfinyl-
and sulfonyl-8H-thieno[2,3-b]pyrrolizin-8-oximino deriv-
atives were synthesized and most of them exhibited bet-
ter anti-tumor activities than cisplatin against Bel-7402
i 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2007, 340, 416–423
Novel Thieno[2,3-b]pyrrolizines
419
(DMSO-d6), d: 2.48 (s, 3H, SCH3), 3.73 (s, 3H, COOCH3), 6.07 (s, 2H,
D2O exchangeable, NH2), 7.75 (d, 2H, J = 8.4 Hz, Ph-2,6-2H), 8.39
(d, 2H, J = 8.4 Hz, Ph-3,5-2H).
and HT-1080 cells. Among them, compound 11N (IC50
=
18.2 lM, 8.2 lM), the most promising compound of all
synthesized derivatives, was 2.5- and 3.3-times more
active than cisplatin (IC50 = 45.2 lM, 26.7 lM), respec-
tively. A preliminary SAR led to identification of several
crucial structural requirements needed to enhance the
effectiveness of alkylthiothienopyrrolizin-8-oximino
derivatives. In particular, our results suggest what the
required determinants should be: (i) the presence of suit-
able dialkylamino ethyloxymethyloxyimino substitu-
ents, especially 4-methyl piperazine in the amino moiety,
at position-8 of thieno[2,3-b]pyrrolizine scaffold; (ii) the
sulfur atom in non-oxidation state. Moreover, being the
first report on 2-methylthio-, sulfinyl-, and sulfonyl-8H-
thieno[2,3-b]pyrrolizin-8-oximino derivatives serving as
anti-tumor agents, these results propose a new sulfur-
containing lead compound in the research and develop-
ment of anti-tumor drugs. More work is needed to ana-
lyze the influence of other substituents, especially at
position-3 on the tripentacyclic core, and studies in this
sense are currently underway.
Methyl 5-methylthio-4-(4-nitrophenyl)-3-(1H-pyrrol-1-
yl)thiophene-2-carboxylate 3
To a solution of 2 (32.4 g, 0.1 mol) in acetic acid 2,5-dimethoxyte-
trahydrofuran (13.2 g, 0.1 mol) was added, the reaction mixture
was stirred at 708C for 4 h, cooled, poured into cold water
(1000 mL), and extracted with methylene chloride. The com-
bined organic phase was washed with water, dried over anhy-
drous sodium sulfate, and was evaporated in vacuo to yield a
solid product (32.9 g, 88%), mp. 110–1128C. MS [MH+] (m/z):
375.0; IR (KBr) cm– 1: 1720.8 (C=O), 1588.5 1548.7 (C=C, C=N),
1516.6 1348.5 (NO2). 1H-NMR (CDCl3), d: 2.51 (s, 3H, SCH3), 3.79 (s,
3H, OCH3), 6.12 (brs, 2H, pyrrol-3,4-2H), 6.52 (brs, 2H, pyrrol-2,5-
2H), 7.78 (d, 2H, J = 8.4 Hz, Ph-2,6-2H), 8.41 (d, 2H, J = 8.4 Hz, Ph-
3,5-2H).
5-Methylthio-4-(4-nitrophenyl)-3-(1H-pyrrol-1-yl)thien-2-
ylpyrrolidine carboxamide 4
A solution of 3 (3.74 g, 0.01 mol) in pyrrolidine (25 mL) was
stirred at 708C for 4 h. After cooling to room temperature, the
resulting mixture was poured into water, the resulting precipi-
tate was filtered, washed with diethyl ether and then was dried;
the slight-yellow solid 4 gave (3.51 g, 85%), m.p 143–1458C. MS
[MH+] (m/z): 414.3; IR (KBr) cm– 1: 1638.8 (C=O), 1602.0 1552.6
(C=C, C=N), 1517.4 1350.2 (NO2). 1H-NMR (CDCl3), d: 1.62 (m, 2H,
pyrrolinyl-2CH), 1.76 (m, 2H, pyrrolinyl-2CH), 2.40 (s, 3H, SCH3),
2.64 (brs, 2H, pyrrolinyl-2CH), 3.50 (m, 2H, pyrrolinyl-2CH), 6.08
(m, 2H, pyrrol-3,4-2CH), 6.48 (m, 2H, pyrrol-2,5-2CH), 7.75 (d, 2H,
J = 8.4 Hz, Ph-2,6-2H), 8.37 (d, 2H, J = 8.4 Hz, Ph-3,5-2H).
Experimental
Chemistry
All melting points were obtained on a Bꢀchi Melting Point B-540
apparatus (Bꢀchi Labortechnik, Flawil, Switzerland) and are
uncorrected. Mass spectra (MS) were taken in ESI mode on Agi-
lent 1100 LC-MS (Agilent, Palo Alto, CA, USA). Proton (1H) nuclear
magnetic resonance spectroscopy were performed using Bruker
ARX-300, 300MHz spectrometers (Bruker Bioscience, Billerica,
MA, USA) with TMS as an internal standard. IR spectra (KBr disks)
were recorded with a Bruker IFS 55 instrument (Bruker). Elemen-
tal analysis was determined on a Carlo-Erba 1106 Elemental
analysis instrument (Carlo Erba, Milan, Italy). Unless otherwise
noted all solvents and reagents were commercially available and
used without further purification.
2-Methylthio-3-(4-nitrophenyl)-8H-thieno[2,3-b]pyrrolizin-
8-one 5
A solution of 4 (4.13 g, 0.01 mol) in phosphorous oxychloride
(25 mL) was stirred at 808C for 2 h. After cooling to room temper-
ature, the resultant mixture was concentrated to give a brown
solid which was filtered, washed with diethyl ether, and dried.
Then, the intermediate was slowly added to a 10% aqueous
sodium hydroxide solution (100 mL) and the mixture was
heated at 608C for 2 h. After cooling, the resulting precipitate
was collected by filtration, washed with water, dried, and recrys-
tallized from ethanol/methanol (2/1) to obtain 5 (2.7 g, 79%) as a
pale white powder, m.p 207-2098C. MS [MH+] (m/z): 343.0; IR (KBr)
cm-1: 2970.0 (CH3), 1686.0 (C=O), 1587.8 1568.8 1549.0 (C=C, C=N),
Methyl 3-amino-5-methylthio-4-(4-nitrophenyl)thiophene-
2-carboxylate 2
1
2-(4-Nitrophenyl)acetonitrile (40.5 g, 0.25 mol) and sodium
hydroxide (20.0 g, 0.5 mol) was added to dry THF (350 mL). After
10 min, carbon disulfide (28.5 g, 0.375 mol) was added in one
portion, the mixture was stirred at room temperature for addi-
tional 30 min. Then cooled to –208C, methyl 2-chloroacetate
(27.3 g, 0.25 mol) in dry THF (50 mL) was added dropwise and
after 1 h, methyl iodide (35.5 g, 0.25 mol) in dry THF (50 mL) was
dropped into the solution. After stirring at –208C for 4 h, the
resulting mixture was poured into water (1000 mL) and
extracted with methylene chloride. The organic phase was dried
over sodium sulfate and evaporated in vacuo to yield a yellow oil.
Recrystallization of the oil from ethyl acetate/hexane (1/1)
obtained 42.9 g (53%) of 2 as pale white powder, mp. 131-1328C.
MS [MH+] (m/z): 325.2; IR (KBr) cm– 1: 3420.5 (NH), 1718.9 (C=O),
1516.4, 1349.8 (NO2). H-NMR (CDCl3), d: 2.61 (s, 3H, SCH3), 6.07
(m, 1H, 6-H), 6.59 (d, 1H, J = 2.9 Hz, 5-H), 6.74 (d, 1H, J = 3.3 Hz, 7-
H), 7.87 (d, 2H, J = 8.5 Hz, Ph-2,6-2H), 8.40 (d, 2H, J = 8.5 Hz, Ph-3,5-
2H); Anal. Calcd. for C16H10N2O3S2: C 56.13, H 2.94, N 8.18 Found:
C 56.21, H 2.92, N 8.23.
8-Hydroxyimino-2-methylthio-3-(4-nitrophenyl)-8H-
thieno[2,3-b]pyrrolizine 6
A mixture of 5 (6.84 g, 0.02 mol) and hydroxylamine hydrochlor-
ide (2.76 g, 0.04 mol) in pyridine (100 mL) was stirred at 908C for
4 h. The mixture was cooled and then evaporated, and the resi-
due was dissolved in methylene chloride (150 mL). The resultant
solution was washed with water, dried over sodium sulfate,
evaporated in vacuo to give the a yellow solid, purified by column
1
1581.6 1540.7 1487.7 (C=C, C=N), 1516.8 1347.6 (NO2). H-NMR
i 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

420
S. Guo et al.
Arch. Pharm. Chem. Life Sci. 2007, 340, 416–423
chromatography on silica gel using ethyl acetate/hexane (1/10)
as eluent. Compound 6 was afforded as a yellow powder (5.14 g,
72%) in a mixture of Z- and E-isomers (Z : E = 70 : 30) not sepa-
7C:
(Z : E = 60 : 40) Yield: 84%; mp. 151–1538C; MS [MH+] (m/z): 455.0;
IR (KBr) cm– 1: 3425.1 (OH), 2952.4 (CH3), 1719.2 (C=O), 1598.1
1558.7 (C=C, C=N), 1516.7 1348.0 (NO2). 1H-NMR (DMSO-d6), Z-iso-
mer d: 1.90 (brs, 4H, pyrrolinyl-3,4-2CH2), 2.57 (s, 3H, SCH3), 3.20
(brs, 2H, pyrrolinyl-2CH), 3.32 (brs, 2H, pyrrolinyl-2CH), 3.56 (brs,
2H, CH2N), 4.60 (s, 2H, OCH2), 6.12 (m, 1H, 6-H), 6.60 (d, 1H, J =
3.4 Hz, 7-H), 6.63 (m, 1H, 5-H), 7.85 (d, 2H, J = 8.4 Hz, Ph-2,6-2H),
8.41 (d, 2H, J = 8.4 Hz, Ph-3,5-2H), E isomer d: 1.90 (brs, 4H, pyrro-
linyl-3,4-2CH2), 2.55 (s, 3H, SCH3), 3.20 (brs, 2H, pyrrolinyl-2CH),
3.32 (br s, 2H, pyrrolinyl-2CH), 3.56 (brs, 2H, CH2N), 4.60 (s, 2H,
OCH2), 6.17 (m, 1H, 6-H), 6.63 (m, 1H, 5-H), 6.82 (brd, 1H, 7-H),
7.85 (d, 2H, J = 8.4 Hz, Ph-2,6-2H), 8.41 (d, 2H, J = 8.4 Hz, Ph-3,5-
2H); Anal. Calcd. for C24H24N4O7S2: C 52.93, H 4.44, N 10.29 Found:
C 53.11, H 4.39, N 10.33.
rated, mp. 148–1518C. MS [MH+] (m/z): 358.0; IR (KBr) cm– 1
:
1
3243.9 (OH), 1596.6, 1558.6 (C=C, C=N), 1518.4 1354.1 (NO2). H-
NMR (CDCl3), Z isomer d: 2.48 (s, 3H, SCH3), 6.08 (m, 1H, 6-H), 6.53
(d, 1H, J = 2.8 Hz, 5-H), 6.55 (d, 1H, J = 3.3 Hz, 7-H), 7.73 (d, 2H, J =
8.6 Hz, Ph-2,6-2H), 8.39 (d, 2H, J = 8.6 Hz, Ph-3,5-2H), E isomer d:
2.43 (s, 3H, SCH3), 6.14 (m, 1H, 6-H), 6.50 (brd, 1H, 5-H), 6.80 (d,
1H, J = 3.3 Hz, 7-H), 7.73 (d, 2H, J = 8.6 Hz, Ph-2,6-2H), 8.38 (d, 2H,
J = 8.6 Hz, Ph-3,5-2H); Anal. Calcd. for C16H11N3O3S2: C 53.77, H
3.10, N 11.76 Found: C 53.65, H 3.12, N 11.81.
General procedure for preparation of compounds
7A-F
The oxime 6 (0.5 g, 1.4 mmol) was dissolved in N,N-dimethylfor-
mamide (DMF) (10 mL) and appropriate aminoalkyl chloride
hydrochloride (1.6 mmol) and potassium carbonate (0.38 g,
2.8 mol) was added to the reaction mixture, which was stirred
for 5 h at 708C. After cooling to room temperature, the resulting
mixture was poured into water, extracted with methylene chlor-
ide and washed with water. The organic phase was dried over
sodium sulfate, and evaporated in vacuo to yield a yellow oil
which was taken up in diethyl ether (50 mL). Oxalic acid (0.13 g,
1.4 mmol) in diethyl ether was added to the resultant solution,
and then the precipitate was filtered, washed with diethyl ether,
and dried to give the desired compounds 7 in a mixture of Z- and
E-isomers not separated.
7D:
(Z : E = 75 : 25) Yield: 72%; mp. 149–1518C; MS [MH+] (m/z): 429.0;
IR (KBr) cm– 1: 3425.9 (OH), 2920.2 (CH2), 1719.9 (C=O), 1598.7
1557.4 1489.2 (C=C, C=N), 1517.4 1347.4 (NO2). 1H-NMR (CDCl3), Z
isomer d: 2.46 (m, 9H, SCH3 and N(CH3)2 ), 2.91 (brt, 2H, CH2N),
4.52 (t, 2H, J = 5.4 Hz, OCH2), 6.07 (m, 1H, 6-H), 6.52 (m, 1H, 5-H),
6.55 (d, 1H, J = 3.2 Hz, 7-H), 7.72 (d, 2H, J = 8.5 Hz, Ph-2,6-2H), 8.38
(d, 2H, J = 8.5 Hz, Ph-3,5-2H), E-isomer d: 2.42 (s, 3H, SCH3), 2.46
(m, 6H, N(CH3)2), 2.91 (brt, 2H, CH2N), 4.52 (t, 2H, J = 5.4 Hz,
OCH2), 6.10 (m, 1H, 6-H), 6.49 (m, 1H, 5-H), 6.71 (m, 1H, 7-H), 7.72
(d, 2H, J = 8.5 Hz, Ph-2,6-2H), 8.38 (d, 2H, J = 8.5 Hz, Ph-3,5-2H);
Anal. calcd. for C22H22N4O7S2: C 50.96, H 4.28, N 10.80 Found: C
51.12, H 4.35, N 10.71.
7E:
7A:
(Z : E = 70 : 30) Yield: 82%; mp. 154–1568C; MS [MH+] (m/z): 469.1;
IR (KBr) cm– 1: 3422.1 (OH), 2929.6 (CH2), 1720.3 (C=O), 1598.2
1557.9 (C=C, C=N), 1518.3 1347.0 (NO2). 1H-NMR (DMSO-d6), Z iso-
mer d: 1.38 (brs, 2H, piperidinyl-4-CH2), 1.51 (brs, 4H, piperidinyl-
3,5-2CH2), 2.53 (m, 7H, piperidinyl-2,6-2CH2 and SCH3), 2.72 (brs,
2H, CH2N), 4.38 (brs, 2H, OCH2), 6.09 (m, 1H, 6-H), 6.56 (brd, 1H, 7-
H), 6.60 (brs, 1H, 5-H), 7.83 (brd, 1H, Ph-2,6-2H), 8.38 (d, 1H, J =
8.1 Hz, Ph-3,5-2H), E-isomer d: 1.38 (brs, 2H, piperidinyl-4-CH2),
1.51 (brs, 4H, piperidinyl-3,5-2CH2), 2.49 (s, 3H, SCH3), 2.53 (m,
4H, piperidinyl-2,6-2CH2), 2.72 (brs, 2H, CH2N), 4.38 (brs, 2H,
OCH2), 6.14 (m, 1H, 6-H), 6.60 (brs, 1H, 5-H), 6.68 (brs, 1H, 7-H),
7.83 (brd, 1H, Ph-2,6-2H), 8.38 (d, 1H, J = 8.1 Hz, Ph-3,5-2H); Anal.
Calcd. for C25H26N4O7S2: C 53.75, H 4.69, N 10.03 Found: C 53.68,
H 4.66, N 10.12.
(Z : E = 65 : 35) Yield: 86%; mp. 198–2008C; MS [MH+] (m/z): 471.0;
IR (KBr) cm– 1: 3420.4 (OH), 2921.5 (CH2), 1719.2 (C=O), 1597.0
1558.2 (C=C, C=N), 1514.8 1346.7 (NO2). 1H-NMR (CDCl3), Z-isomer
d: 2.47 (s, 3H, SCH3), 2.68 (brs, 4H, morpholino-2CH2), 2.92 (s, 2H,
CH2N), 3.79 (s, 4H, morpholino-2CH2), 4.54 (s, 2H, OCH2), 6.07 (m,
1H, 6-H), 6.51 (d, 1H, J = 2.6 Hz, 5-H), 6.55 (d, 1H, J = 3.5 Hz, 7-H),
7.72 (d, 2H, J = 8.6 Hz, Ph-2,6-2H), 8.38 (s, 2H, J = 8.6 Hz, Ph-3,5-
2H), E-isomer d: 2.42 (s, 3H, SCH3), 2.68 (brs, 4H, morpholino-
2CH2), 2.92 (s, 2H, CH2N), 3.79 (s, 4H, morpholino-2CH2), 4.54 (s,
2H, OCH2), 6.11 (m, 1H, 6-H), 6.49 (d, 1H, J = 2.6 Hz, 5-H), 6.69 (d,
1H, J = 3.4 Hz, 7-H), 7.72 (d, 2H, J = 8.6 Hz, Ph-2,6-2H), 8.38 (s, 2H,
J = 8.6 Hz, Ph-3,5-2H); Anal. Calcd. for C24H24N4O8S2: C 51.42, H
4.32, N 9.99 Found: C 51.32, H 4.31, N 10.12.
7F:
(Z : E = 65 : 35) Yield: 75%; mp. 186–1888C; MS [MH+] (m/z): 484.0;
IR (KBr) cm– 1: 3422.1 (OH), 2932.7 (CH2), 1718.4 (C=O), 1598.3
1557.7 (C=C, C=N), 1519.2 1347.2 (NO2). 1H-NMR (CDCl3), Z isomer
d: 2.41 (s, 3H, NCH3), 2.47 (s, 3H, SCH3), 2.66 (brs, 4H, piperazinyl-
4H), 2.76 (brs, 4H, piperazinyl-4H), 2.88 (t, 2H, J = 5.5 Hz, CH2N),
4.48 (t, 2H, J = 5.5 Hz, OCH2), 6.07 (m, 1H, 6-H), 6.51 (d, 1H, J
=2.7 Hz, 5-H), 6.55 (d, 1H, J = 3.5 Hz, 7-H), 7.72 (d, 2H, J = 8.6 Hz,
Ph-2,6-2H), 8.38 (d, 2H, J = 8.6 Hz, Ph-3,5-2H), E isomer d: 2.41 (s,
3H, NCH3), 2.42 (s, 3H, SCH3), 2.66 (br s, 4H, piperazinyl-4H ), 2.76
(brs, 4H, piperazinyl-4H), 2.88 (t, 2H, J = 5.5 Hz, CH2N), 4.48 (t, 2H,
J = 5.5 Hz, OCH2), 6.10 (m, 1H, 6-H), 6.49 (d, 1H, J = 2.5 Hz, 5-H),
6.69 (d, 1H, J = 3.4 Hz, 7-H), 7.72 (d, 2H, J = 8.6 Hz, Ph-2,6-2H), 8.38
(d, 2H, J = 8.6 Hz, Ph-3,5-2H); Anal. Calcd. for C25H27N5O7S2: C
52.34, H 4.74, N 12.21 Found: C 52.21, H 4.72, N 12.28.
7B:
(Z : E = 60 : 40) Yield: 80%; mp. 143-1458C; MS [MH+] (m/z): 457.1; IR
(KBr) cm– 1: 3434.3 (OH), 2939.7 (CH2), 1719.6 (C=O), 1597.8
1557.6 (C=C, C=N), 1516.5 1348.0 (NO2). 1H-NMR (DMSO-d6), Z-iso-
mer d: 1.13-1.24 (m, 6H, -N(CH2CH3)2), 2.57 (s, 3H, SCH3), 3.19 (m,
4H, N(CH2CH3)2), 3.48 (brs, 2H, CH2N), 4.63 (s, 2H, OCH2), 6.12 (m,
1H, 6-H), 6.60 (d, 1H, J = 3.4 Hz, 7-H), 6.64 (d, 1H, J = 2.5 Hz, 5-H),
7.84 (d, 2H, J = 8.7 Hz, Ph-2,6-2H), 8.41 (d, 2H, J = 8.7 Hz, Ph-3,5-
2H), E-isomer d: 1.13–1.24 (m, 6H, N(CH2CH3)2), 2.55 (s, 3H, SCH3),
3.19 (m, 4H, N(CH2CH3)2), 3.48 (brs, 2H, CH2N), 4.63 (s, 2H, OCH2),
6.18 (m, 1H, 6-H), 6.64 (d, 1H, J = 2.5 Hz, 5-H), 6.81 (d, 1H, J =
3.2 Hz, 7-H), 7.84 (d, 2H, J = 8.7 Hz, Ph-2,6-2H), 8.41 (d, 2H, J =
8.7 Hz, Ph-3,5-2H); Anal. Calcd. for C24H26N4O7S2: C 52.37, H 4.79,
N 10.25 Found: C 52.22, H 4.67, N 10.31.
i 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2007, 340, 416–423
Novel Thieno[2,3-b]pyrrolizines
421
2H, J = 5.4 Hz, OCH2), 6.01 (m, 1H, 6-H), 6.52 (d, 1H, J = 2.8 Hz, 5-
H), 6.59 (d, 1H, J = 3.4 Hz, 7-H), 7.72 (d, 2H, J = 8.7 Hz, Ph-2,6-2H),
8.38 (d, 2H, J = 8.7 Hz, Ph-3,5-2H), E-isomer d: 2.65 (m, 4H, mor-
pholino-3,5-2CH2), 2.87 (brt, 2H, CH2N), 2.89 (s, 3H, SO2CH3),
3.74–3.79 (m, 4H, morpholino-2,6-2CH2), 4.51 (t, 2H, J = 5.4 Hz,
OCH2), 6.06 (m, 1H, 6-H), 6.49 (d, 1H, J = 2.7 Hz, 5-H), 6.68 (d, 1H, J
= 3.3 Hz, 7-H), 7.72 (d, 2H, J = 8.7 Hz, Ph-2,6-2H), 8.38 (d, 2H, J =
8.7 Hz, Ph-3,5-2H); Anal. Calcd. for C24H24N4O10S2: C 48.64, H 4.08,
N 9.45 Found: C 48.52, H 4.06, N 9.51.
General procedure for preparation of the compounds
8G-H
Compound 7 (1.0 mmol) was dissolved in glacial acetic acid
(25 mL), and sodium perborate tetrahydrate (0.154 g, 1.0 mmol)
was successively added into the solution which was then stirred
at 25l308C for 3l4 h. Then, the resulting solution was poured
into water and extracted with methylene chloride. The organic
phase was dried over sodium sulfate and evaporated in vacuo to
yield a yellow oil which was converted into the oxalate salt
according to the preparation of compounds 7.
9J:
(Z A 90%) Yield: 89%; mp. 185–1878C; MS [MH+] (m/z): 488.7; IR
(KBr) cm– 1: 3422.2 (OH), 2968.4 (CH2), 1720.8 (C=O), 1600.5
1558.8 1491.0 (C=C, C=N), 1524.5 1348.7 (NO2). 1H-NMR (CDCl3), Z-
isomer d: 1.12 (m, 6H, N(CH2CH3)2), 2.71 (m, 4H, N(CH2CH3)2), 2.92
(m, 5H, SO2CH3, and CH2N), 4.52 (m, 2H, OCH2), 6.10 (dd, 1H, J1 =
3.6 Hz, J2 = 2.8 Hz, 6-H), 6.25 (d, 1H, J = 2.8 Hz, 5-H), 6.61 (d, 1H, J =
3.6 Hz, 7-H), 7.81 (d, 2H, J = 8.7 Hz, Ph-2,6-2H), 8.42 (d, 1H, J =
8.7 Hz, Ph-3,5-2H); Anal. Calcd. for C24H26N4O9S2 : C 49.82, H 4.53,
N 9.68 Found: C 49.73, H 4.51, N 9.59.
8G:
(Z : E = 80 : 20) Yield: 90%; mp. 187–1898C; MS [MH+] (m/z): 486.7;
IR (KBr) cm-1: 3429.5 (OH), 2925.9 (CH2), 1721.3 (C=O), 1599.5
1557.0 (C=C, C=N), 1523.0 1348.7 (NO2). 1H-NMR (CDCl3), Z-isomer
d: 2.63 (m, 4H, morpholino-3,5-2CH2), 2.86 (brt, 2H, CH2N), 2.90
(s, 3H, SOCH3), 3.76 (t, 4H, J = 4.5 Hz, morpholino-2,6-2CH2), 4.53
(t, 2H, J = 5.5 Hz, OCH2), 6.13 (m, 1H, 6-H), 6.51 (d, 1H, J = 2.8 Hz, 5-
H), 6.61 (d, 1H, J = 3.6 Hz, 7-H), 7.75 (d, 2H, J = 8.6 Hz, Ph-2,6-2H),
8.42 (d, 2H, J =8.6 Hz, Ph-3,5-2H), E-isomer d: 2.63 (m, 4H, morpho-
lino-3,5-2CH2), 2.86 (brt, 2H, CH2N), 2.85 (s, 3H, SOCH3), 3.76 (t,
4H, J = 4.5 Hz, morpholino-2,6-2CH2), 4.53 (t, 2H, J =5.5 Hz, OCH2),
6.16 (m, 1H, 6-H), 6.49 (d, 1H, J = 2.7 Hz, 5-H), 6.75 (d, 1H, J =
3.5 Hz, 7-H), 7.75 (d, 2H, J = 8.6 Hz, Ph-2,6-2H), 8.42 (d, 2H, J
=8.6 Hz, Ph-3,5-2H); Anal. Calcd. for C24H24N4O9S2: C 49.99, H 4.20,
N 9.72 Found: C 50.14, H 4.18, N 9.86.
8-(2-Chloroethyloxymethyloxyimino)-2-methylthio-3-
(4-nitrophenyl)-8H-thieno[2,3-b]pyrrolizine 10
To a solution of the oxime 6 (0.5 g, 1.4 mmol) in anhydrous DMF
(10 mL), CH3ONa (0.09 g, 1.68 mmol) in methanol (5 mL) was
added dropwise at 08C. After stirring for 10 min at room temper-
ature, 85% aqueous 1-chloro-2-(chloromethoxy)ethane (0.32 g,
2.1 mmol) was added and the resulting solution was continu-
ously stirred for 1 h at 258C. Then, the mixture was poured into
water and the precipitate was collected by filtration, washed
with water, dried, and purified by column chromatography on
silica gel using ethyl acetate/hexane (1/10) as eluent to obtain 10
as a yellow powder (0.46 g ,73%) (Z : E = 40 : 60), mp. 91–938C; MS
[MH+] (m/z): 449.7, 451.6; IR (KBr) cm– 1: 2922.9 (CH2), 1598.6
1558.1 1489.3 (C=C, C=N), 1520.2 1348.6 (NO2). 1H-NMR (CDCl3), Z-
isomer d: 2.49 (s, 3H, SCH3), 3.66 (m, 2H, CH2Cl), 3.98 (m, 2H,
OCH2), 5.42 (s, 2H, OCH2O), 6.07 (m, 1H, 6-H), 6.50 (m, 1H, 5-H),
6.60 (brd, 1H, 7-H), 7.72 (brd, 2H, Ph-2,6-2H), 8.38 (d, 2H, J =
8.4 Hz, Ph-3,5-2H), E-isomer d: 2.43 (s, 3H, SCH3), 3.66 (m, 2H,
CH2Cl), 3.98 (m, 2H, OCH2), 5.42 (s, 2H, OCH2O), 6.12 (m, 1H, 6-H),
6.50 (m, 1H, 5-H), 6.75 (brd, 1H, 7-H), 7.72 (brd, 2H, Ph-2,6-2H),
8.38 (d, 2H, J = 8.3 Hz, Ph-3,5-2H); Anal. Calcd. for C19H16ClN3O4S2:
C 50.72, H 3.58, N 9.34 Found: C 50.59, H 3.54, N 9.29.
8H:
(Z : E = 40 : 60) Yield: 88%; mp. 167–1698C; MS [MH+] (m/z): 472.9;
IR (KBr) cm– 1: 3438.3 (OH), 2924.2 (CH2), 1719.7 (C=O), 1599.7
1491.7 (C=C, C=N), 1522.4 1348.8 (NO2). 1H-NMR (CDCl3), Z-isomer
d: 1.14 (t, 6H, J =7.0 Hz, N(CH2CH3)2), 2.73–2.81 (m, 4H,
N(CH2CH3)2), 2.89 (s, 3H, SOCH3), 3.01–3.04 (m, 2H, CH2N), 4.54
(m, 2H, OCH2), 6.12 (m, 1H, 6-H), 6.50 (d, 1H, J = 2.7 Hz, 5-H), 6.61
(d, 1H, J = 3.6 Hz, 7-H), 7.75 (d, 2H, J = 8.7 Hz, Ph-2,6-2H), 8.42 (d,
1H, J = 8.7 Hz, Ph-3,5-2H), E-isomer d: 1.14 (t, 6H, J = 7.0 Hz,
N(CH2CH3)2), 2.73–2.81 (m, 4H, N(CH2CH3)2), 2.87 (s, 3H, SOCH3),
3.01–3.04 (m, 2H, CH2N), 4.54 (m, 2H, OCH2), 6.16 (m, 1H, 6-H),
6.48 (d, 1H, J = 2.4 Hz, 5-H), 6.76 (d, 1H, J = 3.6 Hz, 7-H), 7.75 (d, 2H,
J =8.7 Hz, Ph-2,6-2H), 8.42 (d, 1H, J = 8.7 Hz, Ph-3,5-2H); Anal.
Calcd. for C24H26N4O8S2: C 51.24, H 4.66, N 9.96 Found: C 51.36, H
4.69, N 9.89.
General procedure for preparation of the compounds
9I-J
General procedure for preparation of the compounds
11K-O
Compound 7 (1.0 mmol) was dissolved in methanol (25 mL),
sodium wolframate dihydrate (catalyst) and 30% aqueous hydro-
gen peroxide (1.13 g, 10 mmol) was successively dropped into
the solution and stirred at 208C for 2 h. Then the resulting solu-
tion was poured into water and extracted with methylene chlor-
ide. The organic phase was dried over sodium sulfate and evapo-
rated in vacuo to yield a yellow oil which was converted into oxa-
late salt according to the preparation of compounds 7.
To a solution of 10 (1.08 g, 2.4 mmol) in anhydrous DMF (30 mL),
appropriate alkyl amine (3.6 mmol) and sodium hydroxide
(1.92 g, 4.8 mmol) was added, the resultant solution was stirred
for 4 h at 608C. After cooling the mixture was poured into water,
the precipitate was collected by filtration, washed with water,
and dried. The solid was converted into oxalic salt according to
the preparation of compounds 7.
9I:
11K:
(Z : E = 80 : 20) Yield: 79%; mp. 197–1998C; MS [MH+] (m/z): 503.2;
IR (KBr) cm– 1: 3424.6 (OH), 2934.2 (CH2), 1718.6 (C=O), 1609.7
1558.9 (C=C, C=N), 1520.4 1347.3 (NO2). 1H-NMR (CDCl3), Z-isomer
d: 2.65 (m, 4H, morpholino-3,5-2CH2), 2.87 (brt, 2H, CH2N), 2.92
(s, 3H, SO2CH3), 3.74–3.79 (m, 4H, morpholino-2,6-2CH2), 4.51 (t,
(Z : E = 80 : 20) Yield: 80%; mp. 180–1828C; MS [MH+] (m/z): 484.9;
IR (KBr) cm– 1: 3427.8 (OH), 2922.6 (CH2), 1719.6 (C=O), 1598.9
1558.5 1488.9 (C=C, C=N), 1519.2 1347.5 (NO2). 1H-NMR (CDCl3), Z-
isomer d: 1.92 (brs, 4H, pyrrolinyl-3,4-2CH2), 2.50 (s, 3H, SCH3),
2.92 (brs, 4H, pyrrolinyl-2,5-2CH2), 3.00 (m, 2H, CH2N), 4.01 (brt,
i 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

422
S. Guo et al.
Arch. Pharm. Chem. Life Sci. 2007, 340, 416–423
2H, OCH2), 5.40 (s, 2H, OCH2O), 6.07 (m, 1H, 6-H), 6.52 (d, 1H, J =
2.2 Hz, 5-H), 6.58 (d, 1H, J = 3.2 Hz, 7-H), 7.72 (d, 2H, J = 8.7 Hz, Ph-
2,6-2H), 8.38 (d, 1H, J = 8.7 Hz, Ph-3,5-2H), E-isomer d: 1.92 (brs,
4H, pyrrolinyl-3,4-2CH2), 2.43 (s, 3H, SCH3), 2.92 (brs, 4H, pyrro-
linyl-2,5-2CH2), 3.00 (brs, 2H, CH2N), 4.01 (brt, 2H, OCH2), 5.40 (s,
2H, OCH2O), 6.10 (m, 1H, 6-H), 6.50 (m, 1H, 5-H), 6.72 (brd, 1H, 7-
H), 7.72 (d, 2H, J = 8.7 Hz, Ph-2,6-2H), 8.38 (d, 1H, J = 8.7 Hz, Ph-3,5-
2H); Anal. Calcd. for C25H26N4O8S2: C 52.25, H 4.56, N 9.75 Found:
C 52.31, H 4.53, N 9.69.
2H, OCH2), 5.39 (s, 2H, OCH2O), 6.06 (m, 1H, 6-H), 6.51 (m, 1H, 5-
H), 6.58 (d, J = 3.2 Hz, 1H, 7-H), 7.72 (d, 1H, J = 8.5 Hz, Ph-2,6-2H),
8.38 (d, 1H, J = 8.5 Hz, Ph-3,5-2H), E-isomer d: 2.43 (s, 3H, SCH3),
2.61(brs, 4H, morpholino-3,5-2CH2), 2.73 (m, 2H, CH2N), 3.75 (brs,
4H, morpholino-2,6-2CH2), 3.92 (brs, 2H, OCH2), 5.39 (s, 2H,
OCH2O), 6.11 (m, 1H, 6-H), 6.51 (m, 1H, 5-H), 6.74 (d, J = 3.1 Hz, 1H,
7-H), 7.72 (d, 1H, J = 8.5 Hz, Ph-2,6-2H), 8.38 (d, 1H, J = 8.5 Hz, Ph-
3,5-2H); Anal. Calcd. for C25H26N4O9S2: C 50.84, H 4.44, N 9.49
Found: C 51.02, H 4.41, N 9.55.
8-(2-Morpholinoethyloxymethyloxyimino)-
2-methylsulfinyl-3-(4-nitrophenyl)-8H-thieno
[2,3-b]pyrrolizine (Z : E = 45 : 55) 12P
11L:
(Z : E = 90 : 10) Yield: 77%; mp. 177–1798C; MS [MH+] (m/z): 458.8;
IR (KBr) cm– 1: 3432.9 (OH), 2925.1 (CH2), 1719.4 (C=O), 1633.6
1599.2 1558.7 (C=C, C=N), 1518.4 1348.7 (NO2). 1H-NMR (DMSO-
d6), Z isomer d: 2.58 (s, 3H, SCH3), 2.71 (s, 6H, N(CH3)2), 3.23 (brs,
2H, CH2N), 3.94 (brs, 2H, OCH2), 5.40 (s, 2H, OCH2O), 6.13 (m, 1H,
6-H), 6.61 (d, 1H, J = 3.4 Hz, 7-H), 6.65 (d, 1H, J = 2.7 Hz, 5-H), 7.85
(d, 2H, J = 8.5 Hz, Ph-2,6-2H), 8.42 (d, 2H, J = 8.5 Hz, Ph-3,5-2H), E-
isomer d: 2.53 (s, 3H, SCH3), 2.71 (s, 6H, N(CH3)2), 3.23 (brs, 2H,
CH2N), 3.94 (brs, 2H, OCH2), 5.40 (s, 2H, OCH2O), 6.18 (brs, 1H, 6-
H), 6.65 (d, 1H, J = 2.7 Hz, 5-H), 6.73 (brd, 1H, 7-H), 7.85 (d, 2H, J =
8.5 Hz, Ph-2,6-2H), 8.42 (d, 2H, J = 8.5 Hz, Ph-3,5-2H); Anal. Calcd.
for C23H24N4O8S2: C 50.36, H 4.41, N 10.21 Found: C 50.22, H 4.39,
N 10.15.
Compound 12P was prepared from 11O following the procedure
described for 8 and obtained in 85% yield as a yellow solid; mp.
123–1258C; MS [MH+] (m/z): 516.6; IR (KBr) cm– 1: 3419.2 (OH),
2910.2 (CH2), 1719.6 (C=O), 1599.5 1554.9 1492.6 (C=C, C=N),
1521.0 1349.3 (NO2). 1H-NMR (CDCl3), Z isomer d: 2.80 (overlap,
6H, morpholino-2CH2 and CH2N), 2.89 (s, 3H, SOCH3), 3.84 (brs,
4H, morpholino-2CH2), 4.03 (brs, 2H, OCH2), 5.43 (s, 2H,OCH2O),
6.13 (m, 1H, 6-H), 6.52 (m, 1H, 5-H), 6.64 (d, 1H, J = 3.6 Hz, 7-H),
7.75 (d, 2H, J = 8.5 Hz, Ph-2,6-2H), 8.42 (d, 2H, J = 8.5 Hz, Ph-3,5-
2H), E-isomer d: 2.80 (overlap, 6H, morpholino-2CH2 and CH2N),
2.87 (s, 3H, SOCH3), 3.84 (brs, 4H, morpholino-2CH2), 4.03 (brs,
2H, OCH2), 5.43 (s, 2H,OCH2O), 6.18 (m, 1H, 6-H), 6.49 (m, 1H, 5-
H), 6.79 (d, 1H, J = 3.5 Hz, 7-H), 7.75 (d, 2H, J = 8.5 Hz, Ph-2,6-2H),
8.42 (d, 2H, J = 8.5 Hz, Ph-3,5-2H); Anal. Calcd. for C25H26N4O10S2: C
49.50, H 4.32, N 9.24 Found: C 50.06, H 4.30, N 9.13.
11M:
(Z : E = 60 : 40) Yield: 79%; mp. 100–1048C; MS [MH+] (m/z): 498.7;
IR (KBr) cm– 1: 3426.9 (OH), 2922.3 (CH2), 1718.8 (C=O), 1598.6
1557.5 1490.1 (C=C, C=N), 1519.2 1347.6 (NO2). 1H-NMR (CDCl3), Z
isomer d: 1.77 (brs, 6H, piperidinyl-6H), 2.50 (s, 3H, SCH3), 2.88
(overlap, 6H, CH2N and piperidinyl-4H), 4.04 (brs, 2H, OCH2), 5.38
(s, 2H, OCH2O), 6.07 (m, 1H, 6-H), 6.53 (m, 1H, 5-H), 6.58 (m, 1H, 7-
H), 7.72 (d, 1H, J = 8.5 Hz, Ph-2,6-2H), 8.38 (d, 1H, J = 8.5 Hz, Ph-3,5-
2H), E isomer d: 1.77 (brs, 6H, piperidinyl-6H), 2.43 (s, 3H, SCH3),
2.88 (overlap, 6H, CH2N and piperidinyl-4H), 4.04 (brs, 2H, OCH2),
5.38 (s, 2H, OCH2O), 6.12 (m, 1H, 6-H), 6.51 (m, 1H, 5-H), 6.74 (m,
1H, 7-H), 7.72 (d, 1H, J = 8.5 Hz, Ph-2,6-2H), 8.38 (d, 1H, J = 8.5 Hz,
Ph-3,5-2H); Anal. Calcd. for C26H28N4O8S2: C 53.05, H 4.79, N 9.52
Found: C 52.96, H 4.75, N 9.43.
Pharmacology
The MTT cell proliferation assay [18] was used to test the anti-can-
cer activity of all synthesized compounds. The cells were seeded
in RPMI-1640 medium (100 lL) in a 96-well plate at a concentra-
tion of 4000 cells per well. After cultured for 12 h at 378C and
with 5% CO2, cells were incubated with scalar concentrations of
samples for 24 h. MTT was added at a terminal concentration of
5 lg/mL and incubated with cells for 4 h. The formazane crystals
were dissolved in DMSO (100 lL) each well. The optical density
was measured at 492 nm (for absorbance of MTT formazane) and
630 nm (for the reference wavelength). All of the compounds
were tested twice in each of the cell lines. The results expressed
as IC50 (inhibitory concentration 50%) were the averages of two
determinations and calculated by using the Bacus Laboratories
Incorporated Slide Scanner (Bliss) software.
11N:
(Z : E = 70 : 30) Yield: 74%; mp. 192–1948C; MS [MH+] (m/z): 513.7;
IR (KBr) cm– 1: 3435.5 (OH), 3014.5 (C=CH), 1725.2 (C=O), 1599.4
1558.7 (C=C, C=N), 1519.4 1349.3 (NO2). 1H-NMR (DMSO-d6), Z iso-
mer d: 2.58 (s, 3H, SCH3), 2.69 (brs, 7H, piperazinyl-7H), 3.08 (over-
lap, 6H, piperazinyl-4H and CH2N), 3.79 (brs, 2H, OCH2), 5.36 (s,
2H, OCH2O), 6.12 (brs, 1H, 6-H), 6.62 (overlap, 1H, 7-H), 6.64 (over-
lap,1H, 5-H), 7.87 (brd, 2H, Ph-2,6-2H), 8.41 (brd, 2H, Ph-3,5-2H), E
isomer d: 2.53 (s, 3H, SCH3), 2.69 (brs, 7H, piperazinyl-7H), 3.08
(overlap, 6H, piperazinyl-4H and CH2N), 3.79 (brs, 2H, OCH2),
5.36 (s, 2H, OCH2O), 6.14 (brs, 1H, 6-H), 6.64 (overlap,1H, 5-H),
6.72 (brs,1H, 7-H), 7.87 (brd, 2H, Ph-2,6-2H), 8.41 (brd, 2H, Ph-3,5-
2H); Anal. Calcd. for C26H29N5O8S2: C 51.73, H 4.84, N 11.60 Found:
C 52.03, H 4.77, N 11.52.
References
[1] P. Pevarello, M. Villa, Expert Opin. Ther. Patents 2005, 15,
675–703.
[2] Z. Li, Q. Yang, X. Qian, Bioorg. Med. Chem. 2005, 13, 4864–
4870.
[3] Z. Li, Q. Yang, X. Qian, Tetrahedron 2005, 61, 6634–6641.
[4] K. Starcevic, M. Kralj, I. Piantanida, L. Suman, et al., Eur. J.
Med. Chem. 2006, 41, 925–939.
11O:
(Z : E = 65 : 35) Yield: 86%; mp. 113–1158C; MS [MH+] (m/z): 500.7;
IR (KBr) cm– 1: 3429.0 (OH), 2921.0 (CH2), 1717.5 (C=O), 1598.7
1557.3 1489.3 (C=C, C=N), 1519.5 1347.9 (NO2). 1H-NMR (CDCl3), Z-
isomer d: 2.48 (s, 3H, SCH3), 2.61 (brs, 4H, morpholino-3,5-2CH2),
2.73 (m, 2H, CH2N), 3.75 (brs, 4H, morpholino-2,6-2CH2), 3.92 (m,
[5] Z. Zhang , L. Jin, X. Qian, M. Wei, et al., Chembiochem
2007, 8, 113–121.
[6] Q. Li, H. L. Sham, Expert. Opin. Ther. Patents 2002, 12,
1663–1702.
i 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2007, 340, 416–423
Novel Thieno[2,3-b]pyrrolizines
423
[7] C. Rochais, E. Lescot, V. Lisowski, A. Lepailleur, et al., J.
Enzyme Inhib. Med. Chem. 2004, 19, 585–593.
[13] D. Prim, G. Kirsch, Syn. Commun. 1995, 25, 2449–2455.
[14] N. Clauson–Kass, Z. Tyle, Acta Chem. Scand. 1952, 6, 667–
670; ibid 1952, 6, 867–874.
[8] S. Rault, M. Robba, J.-C. Lancelot, H. Prunier, et al., EP
718299. 1996 [Chem. Abstr.] 1996, 125, 142700b.
[15] W. W. Hartman, Org. Synth. Coll. Vol. 1943, 2, 183–183.
[9] I. Baglin, C. Daveu, J.-C. Lancelot, R. Bureau, et al., Bioorg.
Med. Chem. Lett. 2001, 11, 453–457.
[16] L. A. R. Hall, V. C. Stephen, J. H. Burckhalter, Org. Synth.
Coll. Vol. 1963, 4, 333–334.
[10] V. Lisowsiki, C. Enguehared, J.-C. Lancelot, D.-H.
Caignard, et al., Bioorg. Med. Chem. Lett. 2001, 11, 2205–
2208.
[17] P. L. Ferrarini, C. Mori, G. Primofiore, A. Da Settimo, et
al., Eur. J. Med. Chem. 1990, 25, 489–496.
[11] V. Lisowski, S. Leonce, L. Kraus-Berthier, J. Sopkova de Oli-
[18] Method of Pharmacology (Ed.: S. Y. Xu, R. L. Bian, X.
Chen), Public health publishing house, Beijing, 2002, pp.
1784–1786.
veira Santos, et al., J. Med. Chem. 2004, 27, 1448–1464.
[12] K. H. Altmann, Curr. Pharm. Des. 2005, 11, 1595–1613.
i 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com