Synthesis, anticancer and antiviral activities of novel thiopyrano[2,3-d]thiazole-6-carbaldehydes

Article abstract of DOI:10.1080/10426507.2016.1166108

Novel rel-(6R,7R)-2-oxo-7-phenyl-3,5,6,7-tetrahydro-2H-thiopyrano[2,3-d]thiazole-6-carbaldehydes were synthesized via regio- and diastereoselective hetero-Diels-Alder reaction of 5-arylidene-4-thioxo-2-thiazolidinones with acrolein. The synthesized compou

Full text of DOI:10.1080/10426507.2016.1166108

Phosphorus, Sulfur, and Silicon and the Related Elements
ISSN: 1042-6507 (Print) 1563-5325 (Online) Journal homepage: http://www.tandfonline.com/loi/gpss20
Synthesis, anticancer and antiviral activities of
novel thiopyrano[2,3-d]thiazole-6-carbaldehydes
Andrii Lozynskyi, Sergii Golota, Borys Zimenkovsky, Dmytro Atamanyuk,
Andrzej Gzella & Roman Lesyk
To cite this article: Andrii Lozynskyi, Sergii Golota, Borys Zimenkovsky, Dmytro Atamanyuk,
Andrzej Gzella & Roman Lesyk (2016): Synthesis, anticancer and antiviral activities of novel
thiopyrano[2,3-d]thiazole-6-carbaldehydes, Phosphorus, Sulfur, and Silicon and the Related
Elements
To link to this article: http://dx.doi.org/10.1080/10426507.2016.1166108
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thiopyrano[2,3-d]thiazole-6-carbaldehydes
Synthesis, anticancer and antiviral activities of novel thiopyrano[2,3-d]thiazole-6-
carbaldehydes
Andrii Lozynskyia, Sergii Golotaa, Borys Zimenkovskya, Dmytro Atamanyuka, Andrzej Gzellab,
Roman Lesyka,*
aDepartment of Pharmaceutical, Organic and Bioorganic Chemistry, Danylo Halytsky Lviv
National Medical University, 69 Pekarska, Lviv, 79010, Ukraine
bDepartment of Organic Chemistry, Poznan University of Medical Sciences, Grunwaldzka 6,
Poznan 60-780, Poland
*Corresponding author. E-mail: dr_r_lesyk@org.lviv.net (Roman Lesyk)
Abstract
Novel
rel-(6R,7R)-2-oxo-7-phenyl-3,5,6,7-tetrahydro-2H-thiopyrano[2,3-d]thiazole-6-
carbaldehydes were synthesized via regio- and diastereoselective hetero-Diels-Alder reaction of
5-arylidene-4-thioxo-2-thiazolidinones with acrolein. The synthesized compounds were
evaluated for anticancer and antiviral activities by the National Institutes of Health (NIH)
following US NCI and AACF protocols. Anticancer activity screening on NCI60 cell lines
allowed identification of 7-phenyl-2-oxo-7-phenyl-3,5,6,7-tetrahydro-2H-thiopyrano[2,3-
d]thiazole-6-carbaldehyde 3a with the highest level of antimitotic activity against leukemia with
mean GI50/TGI values 1.26/25.22 μM. The screening of antiviral activity lead to identification of
7-(4-methoxyphenyl)-2-oxo-3,5,6,7-tetrahydro-2H-thiopyrano[2,3-d]thiazole-6-carbaldehyde 3b
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with a promising influence on EBV virus (EC50 = 0.07 μM, SI = 3279) and moderate effect on
Herpes simplex virus type 1 and Varicella zoster virus and 7-[4-(benzyloxy)phenyl]-2-oxo-
3,5,6,7-tetrahydro-2H-thiopyrano[2,3-d][1,3]thiazole-6-carbaldehyde 3e with a promising
influence on Hepatitis C virus (EC50 = 12.6 μM, SI = 43.1).
Keywords
hetero-Diels-Alder reaction, thiopyrano[2,3-d]thiazoles, anticancer activity, antiviral activity.
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INTRODUCTION
Thiazolidinone-based molecules are attaractive targets in the rational design of «drug-
like» compounds which possess anti-inflammatory[1], antioxidant[2], antitumor, choleretic,
diuretic[3] and other activities. In addition, 4-thiazolidinones are known as inhibitors of PRL-3
[7]
and JSP-1 phosphatases[4,5], HCV NS3 protease[6], antiapoptic proteins complex Bcl-XL-BH3
TNFα-TNFRc-1 complex[8], Ras farnesyl transferase[9].
,
Among thiazolidinone derivatives, 5-arylidene-4-thiazolidinones are of wide interest due
to their diverse biological activity and clinical applications[3]. However, these compounds often
asigned as the frequent hitters or pan assay interference compounds (PAINS) due to their
Michael acceptors properties[10-13]. Nevertheless, it was found that fixation of a highly active 5-
arylidene-4-thiazolidinone moiety in a structure-related thiopyrano[2,3-d]thiazole core allows to
store of this biophore fragment in a rigid fused system without Michael accepting functionalities.
Thus, in our previous studies we reported about thiopyrano[2,3-d]thiazoles as potential
antitripanosomal[14], antioxidant[15], anticancer[16-20] and anti-inflammatory[21] agents.
In the present work, we described our ongoing research effort in the synthesis of a series
of novel thiopyrano[2,3-d]thiazole-6-carbaldehydes and characterization of these compounds for
antitumor and antiviral activities. In view of these observations it was thought to synthesize these
compounds by the [4+2]‐cyclization with acrolein via hetero-Diels-Alder reaction.
RESULT AND DISCUSSION
Chemistry
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The synthesis of target thiopyrano[2,3-d]thiazoles followed the general pathway outlined
in Scheme 1. Firstly, the starting 5-arylidene-4-thioxo-2-thiazolidinones[21,22] 2a-h were obtained
by Knoevenagel condensation of 4-thioxo-2-thiazolidinone 1 with several aldehydes in the
presence of EDDA as the base catalyst. The reactivity of the sulfur atom at the 4-position at 5-
ylidene-4-thioxo-2-thiazolidinones allows it to be used as a highly active heterodiene component
in hetero-Diels--Alder reactions[23,24]. In this work, we have synthesized several new rel-(6R,7R)-
2-oxo-7-phenyl-3,5,6,7-tetrahydro-2H-thiopyrano[2,3-d]thiazole-6-carbaldehydes
3a-h
by
coupling with acrolein as the dienophile. The heterodiene synthesis was accomplished in glacial
acetic acid and a catalytic amount of hydroquinone, as a side polymerization inhibitor. The
structures and stereochemical features of the synthesized compounds were elucidated by spectral
1
data. Thus, CH2-CH protons of the thiopyran fragment in the H NMR spectra of synthesized
compounds show characteristic patterns of an ABX system. The chemical shifts of these protons
have been assigned a triplet at δ ~2.95-3.02, doublet at δ ~3.12-3.19 and multiplet at δ ~4.7-4.8,
respectively, with corresponding coupling constans of J5,6 = 13.0-13.8 Hz and J6,7 = 4.4-4.7 Hz
which indicated a equatorial-pseudoaxial interaction between the protons at C-6 and C-7. Signals
of the aldehyde group appeared as single-proton singlet in weak magnetic field at δ ~ 9.60-9.75
13
ppm. In the C NMR spectra of the synthesized compounds, the signals observed at δ 171.0--
171.7 are assigned to the carbonyl group (C = O). The signals of carbon of the aldehyde group
were observed at δ 200.5--202.6. Interestingly, similar spectral features were observed for acrylic
acids and their esters as dienophiles in the hetero-Diels-Alder reaction[25]
.
The structure of the synthesized rel-(6R,7R)-7-(4-isopropylphenyl)-2-oxo-3,5,6,7-
tetrahydro-2H-thiopyrano[2,3-d][1,3]thiazole-6-carbaldehyde (3h) was confirmed by X-ray
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crystallographic analysis reaction (Supplemental Materials, Tables S 1-S 4)[26]. In the crystal
structure of 3h there are twelve symmetry-independent molecules in the asymmetric part of the
unit cell. Displacement ellipsoids of the atoms of the p-isopropylphenyl residues in the molecules
are increased. Simultaneously, some of the isopropyl moieties are disordered. The symmetry-
independent molecules differ in terms of conformation. However, the differences mainly relate to
the arrangement of isopropyl moiety.
In all the molecules the tetrahydrothiopyran ring is in a half-chair conformation. The
hydrogen atoms at the stereogenic C7 and C8 centers have a cis axial-pseudoequatorial
orientation. The torsion angles H7--C7--C8--H8 for the twelve symmetry-independent molecules
are in the range of 43  48° and reveal a synclinal (+sc) conformation for atoms H(C7) and
H(C8). The phenyl group is approximately perpendicular to the least squares plane of the
dihydrothiopyran ring; the dihedral angles for symmetry-independent molecules A -- L are in the
range of 87.4(3)  89.3(3)°. The C4C9 bond lengths that are in the range of 1.332(14) -
1.352(14) (molecules A -- L) confirm the presence of a double bond between these atoms. It is
worth noting that the geometry of 2-oxo-7-phenyl-3,5,6,7-tetrahydro-2H-thiopyrano[2,3-
d]thiazole molecule fragment in 3h is close to those observed in two related structures (refcodes:
LOBQEA and QEDBUY) found in Cambridge Structural Database, Version 5.37 [27]
.
Biological activity
In vitro evaluation of the anticancer activity
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Anticancer assays of the compounds 3a-c, 3e, 3g were performed according to the US
NCI protocol, as described elsewhere [28-31]. Some synthesized compounds 3b, 3e were submitted
and evaluated against the three human tumor cell lines panel, consisting of NCI-H460 (non-small
cell lung cancer), MCF7 (breast cancer), and SF-268 (CNS cancer) cell lines. Thus, the
substances which reduced the growth of the cell lines to 32% or less were passed on for
evaluation in the full panel of 60 human tumor cells. As a result these compounds successfully
passed the pre-screening phase. On the other hand, compounds 3a, 3g were evaluated at the
single concentration of 105M against the full panel. The human tumor cell lines were derived
from nine different cancer types: leukemia, melanoma, lung, colon, CNS, ovarian, renal,
prostate, and breast cancers. The results for each compound are reported as the percent growth of
treated cells and compared to untreated control cells. Range of growth (%) showed the lowest
and highest growth that was found among different cancer cell line. The synthesized compounds
3a, 3g displayed significant activity in the vitro screen on tested cell lines. Thus, compound 3a
was highly active on renal cancer UO-31 cell line (GP = -1.20%) and compound 3g was highly
active on non-small cell lung cancer NCI-H522 cell line (GP = -2.71%). The results of the
primary screening are shown in Table S 5 (Supplemental).
Finally, the previously selected compound 3a, 3b, 3e as well as compound 3c without the
preliminary prescreening stage were tested in advanced assay against a full panel of about sixty
[27-30]
tumor cell lines at 10-fold dilutions of five concentrations (100, 10, 1, 0.1 and 0.01 μM)
.
All the tested compounds showed a broad spectrum of growth inhibition activity against human
tumor cells with average GI50/TGI50 values 33.64/65.27 (3a), 39.21/82.59 (3b), 46.71/89.79 (3c)
and 17.65/46.41 (3e). Selectivity pattern analysis of cell lines by disease origin can definitely
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affirm selective action of compounds 3a, 3b, 3c and 3e on leukemia cell lines. These compounds
appeared to be most active against selected individual cell lines with the GI50 varying from 1.86
to 11.89. Compound 3a was found to be a highly active inhibitor of the colon cancer line HT29,
ovarian cancer line OVCAR-3, prostate cancer line DU-145, breast cancer line HS 578T.
Compound 3b showed selectivity on renal cancer cell line RXF 393, breast cancer cell line
MCF7 and compound 3e was highly active on melanoma cell line SK-MEL-5. The dose-
dependent assay 60 cell line assay is summarized in Tables S 6 and S 7 (Supplemental
Materials).
Empirical SAR study revealed that the presence of substituents in phenyl ring in position
7 of thiopyrano[2,3-d]thiazole core isn’t favorable for anticancer potency. Interestingly, the
introduction of bulky groups in para-position in phenyl ring in position 7 of thiopyrano[2,3-
d]thiazole core is more tolerated than small lipophiles in position 3’ and 4’.
Evaluation of antiviral activity
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The synthesized compounds 3b, 3d-h were evaluated for antiviral activity according
antimicrobial acquisition and coordinating facility (AACF) screening program [32-34]. The
obtained results are summarized in Table S 8 (Supplemental materials). Among tested
compounds 3b, 3e was found to be the most active. Regarding Epstein--Barr virus (EBV), it was
found that compound 3b showed excellent activity (EC50 = 0.07 μM, SI = 3279) by VCA Elisa
test and possess moderate activity against Herpes simplex virus 1 (SI = 31) and Varicella zoster
virus (SI = 34) by viral CPE test. Importingly, compound 3e with their EC50 values in the range
of 12.66 μM and SI equal to 43.1 was more active than other derivatives against Hepatitis C
virus. In addition, the moderate activity exhibited compound 3g against Varicella zoster virus
(VZV) in HFF cell culture with EC50 values in the range of 23.42 μM and selective index (SI) of
27.2.
EXPERIMENTAL
Chemistry
All materials were purchased from commercial sources and used without purification.
Melting points were measured in open capillary tubes and were uncorrected. The elemental
analyses (C, H, N) were performed using the Perkin--Elmer 2400 CHN analyzer and were within
1
13
0.4% of the theoretical values. The H- and C-NMR spectra were recorded on Varian Gemini
400 MHz or Bruker 125 MHz for frequencies 100 MHz in DMSO-d6 using tetramethylsilane as
an internal standard. Chemical shifts are reported in ppm units with use of δ scale. The purity of
all obtained compounds was checked by TLC.
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The starting compounds: 2,4-thiazolidinedione[35], 4-thioxo-2-thiazolidinone[23] were
obtained according to methods described previously. 5-Arylidene-4-thioxo-2-thiazolidinones
(2a-h) were prepared under Knoevenagel condensation condition: A mixture of 4-thioxo-2-
thiazolidinone (10 mmol), appropriate substituted benzaldehyde (10 mmol) in the ethanol
medium in the presence of catalytic amount of EDDA was boiled for 10 min. Obtained solid
product were collected by filtration and used without further purification. The Supplemental
Materials contains sample 1H and 13C NMR spectra for products 3a-h (Figures S 1 -- S 16)
General procedure of hetero-Diels-Alder reaction affording 3a-3h
A mixture of appropriate 5-arylidene-4-thioxo-2-thiazolidinone (10 mmol) and acrolein (11
mmol) was refluxed for 1 h with a catalytic amount of hydroquinone (2--3 mg) in glacial acetic
acid (10 mL), then left overnight at room temperature. The precipitated crystals were filtered off,
washed with methanol (5--10 mL), and recrystallized from ethanol (10--15 mL).
rel-(6R,7R)-7-Phenyl-2-oxo-3,5,6,7-tetrahydro-2H-thiopyrano[2,3-d][1,3]thiazole-6-
1
carbaldehyde (3a). Yield 65%, mp 186-187°C. H NMR (400 MHz, DMSO-d6): δ 2.95 (t, J =
13.4 Hz, 1H, 5-H), 2.95 (dd, J = 2.4, 13.4 Hz, 1H, 5-H), 3.40 (m, 1H, 6-H), 4.71 (d, J = 4.5 Hz,
1H, 7-H), 7,15-7,35 (m, 5Н, arom.), 9.74 (s, 1Н, СНО), 11.48 (s, 1Н, NH). 13C NMR (100 MHz,
DMSO-d6): δ 35.9 (5-C), 44.1 (7-C), 55.6 (6-C), 109.9, 114.5, 121.3, 130.4, 145.4, 154.8, 171.3
(C = O), 201.7 (CHO). Calcd. for C13H11NO2S2: C, 56.30; H, 4.00; N, 5.05; Found: C, 56.32; H,
4.01; N, 5.04%.
rel-(6R,7R)-7-(4-Methoxyphenyl)-2-oxo-3,5,6,7-tetrahydro-2H-thiopyrano[2,3-d]thiazole-6-
1
carbaldehyde (3b). Yield 74%, mp 207-208°C. H NMR (400 MHz, DMSO-d6): δ 3.03 (t, J =
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13.0 Hz, 1Н, 5-Н), 3.12 (d, J = 12.3 Hz, 1Н, 5-Н), 3.28 (m, 1Н, 6-Н), 3.86 (s, 3Н, ОСН3), 4.60
(d, J = 4.5 Hz, 1Н, 7-Н), 6.82 (d, J = 8.8 Hz, 2Н, arom.), 7.11 (d, J = 8.8 Hz, 2Н, arom.), 9.72 (s,
13
1Н, СНО), 11.22 (s, 1Н, NH). C NMR (100 MHz, DMSO-d6): δ 22.2 (5-C), 38.3 (7-C), 51.6
(ОСН3), 55.6 (6-C), 105.5, 114.5, 121.3, 130.3, 130.4, 131.9, 159.2, 171.3 (C = O), 201.7
(CHO). Calcd. for C14H13NO3S2: C, 54.70; H, 4.26; N, 4.56; Found: C, 54.72; H, 4.24; N, 4.54%.
rel-(6R,7R)-7-(3,4-Dimethoxyphenyl)-2-oxo-3,5,6,7-tetrahydro-2H-thiopyrano[2,3-
d]thiazole-6-carbaldehyde (3c). Yield 56%, mp 197-198°C. 1H NMR (400 MHz, DMSO-d6): δ
3.04 (t, J = 13.0 Hz, 1Н, 5-Н), 3.16 (d, J = 12.3 Hz, 1Н, 5-Н), 3.70 (s, 3Н, ОСН3), 3.72 (s, 3Н,
ОСН3), 4,01 (m, 1Н, 6-Н), 4.52 (d, J = 4.5 Hz, 1Н, 7-Н), 6.69 (d, J = 8.2 Hz, 1Н, arom.), 6.71 (s,
13
1Н, arom.), 6.85 (d, J = 8.2 Hz, 1Н, arom.), 9.45 (s, 1Н, СНО), 11.44 (s, 1Н, NH). C NMR
(100 MHz, DMSO-d6): δ 23.7 (5-C), 38.3 (7-C), 49.4 (ОСН3), 52.4 (ОСН3), 55.6 (6-C), 109.9,
114.5, 124.5, 129.8, 131.9, 134.7, 141.6, 162.0, 171.7 (C = O), 202.1 (CHO). Calcd. for
C15H15NO4S2: C, 53.40; H, 4.48; N, 4.15; Found: C, 53.42; H, 4.47; N, 4.14%.
rel-(6R,7R)-7-{2-[(2-Chlorobenzyl)oxy]-5-nitrophenyl}-2-oxo-3,5,6,7-tetrahydro-2H-
1
thiopyrano[2,3-d][1,3]thiazole-6-carbaldehyde (3d). Yield 57%, mp 237-238°C. H NMR
(400 MHz, DMSO-d6): δ 3.19 (dd, J = 2.4, 13.8 Hz, 1Н, 5-Н), 3.34 (m, 1Н, 5-Н), 3.44 (dd, J =
4.8, 13.8 Hz, 1Н, 6-H), 4.75 (d, J = 4.4 Hz, 1Н, 7-Н), 5.33 (d, J = 12.1 Hz, 1Н, СН2),5.38 (d, J =
12.1 Hz, 1Н, СН2), 7.38-7.44 (m, 3Н, arom.), 7.52 (d, J = 7,2 Hz, 1Н, arom.), 7.61 (d, J = 7.2
Hz, 1Н, arom.), 7.89 (s, 1Н, arom.), 8.26 (d, J = 8.9 Hz, 2Н, arom.), 9.63 (s, 1Н, СНО), 11.53 (s,
1Н, NH). 13C NMR (100 MHz, DMSO-d6): δ 32.4 (5-C), 32.7 (7-C), 49.2 (6-C), 68.8 (СН2),
102.2, 113.4, 122.4, 125.4, 125.8, 128.0, 130.1, 130.5, 130.8, 131.6, 133.0, 133.8, 141.6, 160.7,
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171.0 (C = O), 200.5 (CHO). Calcd. for C20H15ClN2O5S2: C, 51.89; H, 3.27; N, 6.05; Found: C,
51.87; H,3.28; N, 6.04%.
rel-(6R,7R)-7-[4-(Benzyloxy)phenyl]-2-oxo-3,5,6,7-tetrahydro-2H-thiopyrano[2,3-
1
d][1,3]thiazole-6-carbaldehyde (3e). Yield 75%, mp 218-219°C. H NMR (400 MHz, DMSO-
d6): δ 3.02 (t, J = 13.0 Hz, 1Н, 5-Н), 3.13 (dd, J = 2.0, 13.0 Hz, 1Н, 5-Н), 3.32 (m, 1Н, 6-Н),
4.69 (d, J = 4.7 Hz, 1Н, 7-Н), 5.05 (d, J = 8.4 Hz, 2Н, СН2), 7.11 (d, J = 8.4 Hz, 2Н, arom.),
13
7.39-7.43 (m, 5Н, arom.), 9.75 (s, 1Н, СНО), 11,48 (s, 1Н, NH). C NMR (100 MHz, DMSO-
d6): δ 28.8 (5-C), 32.4 (7-C), 49.2 (6-C), 69.9 (СН2), 113.4, 123.2, 128.0, 130.5, 132.2, 133.0,
133.8, 141.6, 149.9, 163.4, 171.0 (C = O), 200.5 (CHO). Calcd. for C20H17NO3S2: C, 62.64; H,
4.47; N, 3.65; Found: C, 62.63; H, 4.48; N, 3.64%.
rel-(6R,7R)-7-[4-(Difluoromethoxy)-3-methoxyphenyl]-2-oxo-3,5,6,7-tetrahydro-2H-
thiopyrano[2,3-d][1,3]thiazole-6-carbaldehyde (3f). Yield 55%, mp 196-197°C. 1H NMR (400
MHz, DMSO-d6): δ 3.02 (t, J = 13.4 Hz, 1Н, 5-Н), 3.13 (d, J = 13.4 Hz, 1Н, 5-Н), 3.32 (m, 1Н,
6-Н), 3.82 (s, 3Н, OСН3), 4.34 (d, J = 4.7 Hz, 1Н, 7-Н), 6.68 (s, 1H, OCHF2), 6.76 (d, J = 8.7
Hz, 1H, arom.), 6.88 (s, 1H, OCHF2), 7,11-7.11 (m, 2H, arom.), 9.60 (s, 1Н, СНО), 11.33 (s,1Н,
NH). 13C NMR (100 MHz, DMSO-d6): δ 23.5 (5-C), 37.9 (7-C), 51.9 (ОСН3), 56.5 (6-C), 104.2,
113.6, 117.3, 120.8, 121.2, 121.7, 141.3, 151.3, 171.1 (C = O), 200.7 (CHO). Calcd. for
C15H13F2NO4S2: C, 48.25; H, 3.51; N, 3.75; Found: C, 48.23; H, 3.52; N, 3.74%.
rel-(6R,7R)-7-(4-tert-Butylphenyl)-2-oxo-3,5,6,7-tetrahydro-2H-thiopyrano[2,3-
1
d][1,3]thiazole-6-carbaldehyde (3g). Yield 72%, mp 226-227°C. H NMR (400 MHz, DMSO-
d6): δ 1.25 (c, 9Н, 3СН3), 2,96 (t, J = 13.4 Hz, 1Н, 5-Н), 3.19 (d, J = 13.4 Hz, 1Н, 5-Н), 3.40 (m,
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1Н, 6-Н), 4.69 (d, J = 4.6 Hz, 1Н, 7-Н), 7.34 (d, J = 8.3 Hz, 2Н, arom.), 7.38 (d, J = 8.3 Hz, 2Н,
13
arom.), 9.76 (s, 1Н, СНО), 11.50 (s, 1Н, NH). C NMR (100 MHz, DMSO-d6): δ 31.7 (СН3),
31.8 (5-C), 34.8, 38.6, 105.2, 121.4, 126.0, 129.0, 137.1, 150.5, 171.3 (C = O), 201.7 (CHO).
Calcd. for C17H19NO2S2: C, 61.23; H, 5.74; N, 4.20; Found: C, 61.22; H,5.72; N, 4.21%.
rel-(6R,7R)-7-(4-Isopropylphenyl)-2-oxo-3,5,6,7-tetrahydro-2H-thiopyrano[2,3-
1
d][1,3]thiazole-6-carbaldehyde (3h). Yield 70%, mp 211-212°C. H NMR (400 MHz, DMSO-
d6): δ 1.20 (d, J = 6.9 Hz, 6Н, 2*СН3), 2.86 (m, 1Н, СН), 3.02 (t, J = 13.4 Hz, 1Н, 5-Н), 3.13 (d,
J = 13.4 Hz, 1Н, 5-Н), 3.32 (m, 1Н, 6-Н), 4.65 (d, J = 4.7 Hz, 1Н, 7-Н), 7.09 (d, J = 8.8 Hz, 2Н,
13
arom.), 7.15 (d, J = 8.8 Hz, 2Н, arom.), 9.73 (s, 1Н, СНО), 11.33 (s,1Н, NH). C NMR (100
MHz, DMSO-d6): δ 22.2 (СН3), 34.9 (5-C), 49.4, 51.6, 58.5, 114.5, 121.3, 130.8, 132.1, 160.2,
171.3 (C = O), 202.6 (CHO). Calcd. for C16H17NO2S2: C, 60.16; H, 5.36; N, 4.38; Found: C,
60.15; H,5.37; N, 4.37%.
Primary anticancer assay.
Primary anticancer assay was performed on a panel of approximately sixty human tumor
cell lines derived from nine neoplastic diseases, in accordance with the protocol of the Drug
Evaluation Branch, National Cancer Institute, Bethesda[27-30]
.
Methods for evaluation of antiviral activity.
Primary antiviral assay was performed at a biodefense virus’s panel and a respiratory
virus’s panel [Flu A (H5N1), Flu B, SARS] with a protocol of the NIAID’s antimicrobial
acquisition and coordinating[31-33]
.
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CONCLUSION
We have achieved a convenient protocol for the synthesis of thiopyrano[2,3-d]thiazole-6-
carbaldehydes via regio- and diastereoselective hetero-Diels-Alder reaction of 5-arylidene-4-
thioxo-2-thiazolidinone with acrolein. The synthesized compounds ware assessed for their
antitumor and antiviral capacities. The preliminary results allowed to identify the most active
compound 3a with mean GI50/TGI50 values 33.64/65.27 μM in the NCI 60 cell-line assay with
certain sensitivity profile towards the leukemia SR cell line (GI50/TGI values 0.89/6.88 μM). The
7-(4-methoxyphenyl)-2-oxo-7-phenyl-3,5,6,7-tetrahydro-2H-thiopyrano[2,3-d]thiazole-6-
carbaldehyde 3b (EC50 = 0.07 μM, SI = 3279) has shown the best antiviral activity against
Epstein--Barr virus in Daudi cell cultures and compound 3e (EC50 = 12.66 μM, SI = 43.1) was
the most active against Hepatitis C virus than the other derivatives. Overall, the biological tests
revealed the necessity for further investigations the antitumor and antiviral potential of tested
compounds for the construction of novel chemical entities with better pharmacological profiles.
ACKNOWLEDGEMENTS
The authors are grateful to Dr. V. L. Narayanan from Drug Synthesis and Chemistry Branch,
National Cancer Institute, Bethesda, MD, USA, for in vitro evaluation of anticancer activity and
to Dr. Chris Tseng, from Division of Microbiology and Infectious Diseases NIAID/NIH,
Bethesda, USA, for in vitro evaluation of antiviral activity.
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REFERENCES
1.Geronikaki, A. A.; Lagunin, A. A.; Hadjipovlou-Litina, D. I.; Eleftheriou, P. T.;
Filimonov, D. A.; Poroikov, V. V.; Alam, I.; Saxena A. K. J. Med. Chem. 2008, 51, 1601-
1609.
2.Shin, M. H.; Ke, F. Y. Bioorg. Med. Chem. Lett. 2004, 12, 4633-4643.
3.Lesyk, R. B.; Zimenkovsky, B. S. Curr. Org. Chem. 2004, 8, 1547–1577.
4.Cutshall, N. S.; O’Day, C.; Prezhdo, M. Bioorg. Med. Chem. Lett. 2005, 15, 3374–3379.
5.Ahn, J. H.; Kim, S. J.; Park, W. S.; Cho, S. Y.; Ha, J. D.; Kim, S. S.; Kang, S. K.; Jeong,
D. G.; Jung, S.-K.; Lee, S.-H.; Kim, H. M.; Park, S. K.; Lee, K. H.; Lee, C. W.; Ryu, S. E.;
Choi, J.-K. Bioorg. Med. Chem. Lett. 2006, 16, 2996–2999.
6.Sing, W. T.; Lee, C. L.; Yeo, S. L.; Lim, S. P.; Sim, M. M. Bioorg. Med. Chem. Lett. 2001,
11, 91–94.
7.Liu, W.; Bulgaru, A.; Haigentz, M.; Stein, C.A.; Perez-Soler, R.; Mani. S. Curr. Med.
Chem.: Anti-Cancer Agents. 2003, 3, 217–223.
8.Carter, P. H.; Scherle, P. A.; Muckelbauer, J. A.; Voss, M. E.; Liu, R.-Q.; Thompson, L.
A.; Tebben, A. J.; Solomon, K. A.; Lo, Y. C.; Li, Z.; Strzemienski, P.; Yang, G.;
Falahatpisheh, N.; Xu, M.; Wu, Z.; Wu, Z.; Farrow, N. A.; Ramnarayan, K.; Wang, J.;
Rideout, D.; Yalamoori, V.; Domaille, P.; Underwood, D. J.; Trzaskos, J. M.; Friedman, S.
M.; Newton, R. C.; Decicco, C. P. Proc. Natl. Acad. Sci. U.S.A. 2001, 98, 11879-11884.
14
ACCEPTED MANUSCRIPT

ACCEPTED MANUSCRIPT
9.Lee, J.; Kim, J.; Koh, J. S.; Chung, H. H.; Kim, K. H. Bioorg. Med. Chem. Lett. 2006, 16,
1954–1956.
10. Tomasic, T.; Masic, L. P. Expert Opin. Drug Discov. 2012, 7, 549-560.
11. Baell, J. B. Future Med. Chem. 2010, 2, 1529-1546.
12. Baell, J. B.; Holloway, G. A. J. Med. Chem. 2010, 53, 2719-2740.
13. Mendgen, T.; Steuer, C.; Klein, C. D. J. Med. Chem. 2012, 55, 743-753.
14. Zelisko, N.; Atamanyuk, D.; Vasylenko, O.; Grellier, P.; Lesyk, R. Bioorg. Med. Chem.
Lett. 2012, 22, 7071-7074.
15. Lozynskyi, A. V.; Kaminskyy, D. V.; Romanchyshyn, K. B.; Semenciv, N. G.; Ogurtsov,
V.V.; Lesyk, R. B. Biopolym. Cell. 2015, 31, 131-137.
16. Lesyk, R. B.; Zimenkovsky, B. S.; Kaminskyy, D. V.; Kryshchyshyn, A. P.; Havrylyuk,
D. Ya.; Atamanyuk, D.V.; Subtel’na, I. Yu.; Khyluk, D. V. Biopolym. Cell. 2011, 27, 107–
117.
17. Lesyk R.; Zimenkovsky B.; Atamanyuk D.; Jensen F.; Kiec-Kononowicz K.; Gzella A.
Bioorg. Med. Chem. 2006, 14, 5230–5240.
18. Lozynskyi, A.; Zimenkovsky, B.; Lesyk, R. Sci. Pharm. 2014, 82, 723-733.
19. Atamanyuk, D.; Zimenkovsky, B.; Lesyk R. J. Sulfur Chem. 2008, 29, 151-162.
15
ACCEPTED MANUSCRIPT

ACCEPTED MANUSCRIPT
20. Kaminskyy, D.; Vasylenko, O.; Atamanyuk, D.; Gzella, A.; Lesyk R. Synlett. 2011, 10,
1385–1388.
21. Lozynskyi, A.; Zimenkovsky, B.; Nektegayev, I.; Lesyk, R. Heterocycl. Commun. 2015,
21, 55-59.
22. Lozynskyi, A.; Zimenkovsky, B.; Gzella, A.; Lesyk, R. Synth. Commun. 2015, 45, 2266-
2270.
23. Komaritsa, I. D.; Baranov, S. N.; Grishchuk, A. P. Khimiya Geterotsiklicheskikh
Soedinenii. 1967, 3, 533-534.
24. Metwally, N. H. J. Sulfur Chem. 2014, 35, 528-537.
25. Zelisko, N.; Atamanyuk, D.; Vasylenko,O.; Bryhas,A.; Matiychuk,V.; Gzella, A.; Lesyk,
R. Tetrahedron. 2013, 70, 720-729.
26. The supplementary crystallographic data of 3h have been deposited at the Cambridge
Crystalographie Data Centre (CCDC) as supplementary publication CCDC1445170.
Copies of the data can be obtained, free of charge, on application to CCDC, 12 Union
Road, Cambridge CB2 1EZ, UK, (fax: +44-(0)1223-336033 or e-mail:
deposit@ccdc.cam.ac.uk).
27. Allen, F. H. Acta Cryst. 2002, B58, 380–388.
28. http://dtp.nci.nih.gov
29. Boyd, M. R.; Paull, K. D. Drug Dev Res. 1995, 34, 91-109.
16
ACCEPTED MANUSCRIPT

ACCEPTED MANUSCRIPT
30. Shoemaker,R. H. Nat. Rev. Cancer. 2006, 6, 813–823.
31. Monks, A.; Scudiero, D.; Skehan, P.; Shoemaker, R.; Paull, K.; Vistica, D.; Hose, C.;
Langley, J.; Cronise, P.; Vaigro-Wolff A.; Gray-Goodrich, M.; Campbell, H.; Mayo, J.;
Boyd, M. J. Nat. Cancer Inst. 1991, 83, 757–766.
32. http://niaid-aacf.org
33. Huffman, J. H.; Sidwell, R. W.; Barnard, D. L.; Morrison, A.; Otto,M. J.; Hill, C.L.;
Schinazi, R. F. Antiv. Chem. Chemother. 1997, 8, 75-83.
34. Sidwell, R. W.; Smee, D. F. Antiviral Res. 2000, 48, 1-16.
35. Turkevych, N. M.; Vvedenskij V. M.; Petlichnaya L. P. Method of obtaining
pseudothiohydantoin and thiazolidinedione-2,4. Ukr. Khim. Zh. (Russ. Ed.) 1961, 27, 680-
681.
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CF3
Cl
O
H
H
H
Me
S
O
H
N
H
O
H
S
N
N
S
H
N
H
O
N
H
S
O
S
O
NH
H
H
O
S
H
S
H
NH
O
O
H
MeO
S
Me
HO
MeO
HO
A. Lozynskyi et al., 2014 [17] D. Atamanyuk et al., 2007 [18]
N. Zelisko et al., 2012 [13]R. Lesyk et al., 2006 [16]
Figure 1. Structures of biologically active thiopyrano[2,3-d]thiazoles.
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Figure 2. X-ray crystal structure (ORTEP plot) of one of twelve symmetry-independent
molecules of 3h and the atom-labeling scheme.
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2a/3a R = H, 75/65%
2b/3b R = 4-MeO, 80/74%
2c/3c R = 3,4-(MeO)2, 78/56%
O
R
CHO
O
H
N
S
S
S
H
O
H
P2S5
H
H
N
H
H
H
N
N
O
2d/3d R = 2-(2-Cl-BzO)-5-NO2, 72/57%
2e/3e R = 4-BzO, 74%/75%
2f/3f R = 3-OMe-4-OCHF2, 72/55%
S
a
O
b
c
S
O
S
O
S
1
3a-h
R
2g/3g R = 4-(CH3)3C, 77/72%
2h/3h R = 4-(CH3)2CH, 75/70%
2a-h
R
Scheme 1. Synthesis of rel-(6R,7R)-2-oxo-7-phenyl-3,5,6,7-tetrahydro-2H-thiopyrano[2,3-
d]thiazole-6-carbaldehydes. Reagents, conditions and yields: (a) dioxane, reflux 3h, 60%; (b)
EDDA, EtOH, reflux 10 min; (c) AcOH, reflux 1h.
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