Synthesis and leukemia cell growth inhibition of a series of 1,3-dithiazolylbenzene derivatives

Article abstract of DOI:10.1135/cccc20041491

By a slightly modified Hantzsch thiazole synthesis either 1,3-bis[(thiazol-2-yl)amino]benzene derivatives 2 or 1,3-bis[2-iminothiazol- 3(2H)-yl]benzene derivatives 3 were exclusively obtained. The compounds can be distinguished by NMR spectroscopy. Compounds 2a-2d and 3a-3d were evaluated for their potential antitumor activity, DNA interaction, and for their activity against DNA and RNA viruses and against HIV-1 and HIV-2.

Full text of DOI:10.1135/cccc20041491

1
,3-Dithiazolylbenzene Derivatives
1491
SYNTHESIS AND LEUKEMIA CELL GROWTH INHIBITION OF
A SERIES OF 1,3-DITHIAZOLYLBENZENE DERIVATIVES
a1
a2
b1
Irene M. LAGOJA , Koen NAUWELAERTS , Christine BAL-MAHIEU ,
a3
b2
a4,
Michela PASQUALINI , Christian BAILLY and Piet HERDEWIJN
*
a
Laboratory of Medicinal Chemistry, Rega Institute for Medical Research,
Katholieke Universiteit Leuven, Minderbroedersstraat 10, B-3000 Leuven, Belgium;
1
2
3
e-mail: irene.lagoja@rega.kuleuven.ac.be, koen.nauwelaerts@rega.kuleuven.ac.be,
4
mich.pas@hotmail.com, piet.herdewijn@rega.kuleuven.ac.be
b
INSERM U-524 et Laboratoire de Pharmacologie Antitumorale du Centre Oscar Lambret, IRCL,
1
2
5
9045 Lille, France; e-mail: mahieu@lille.inserm.fr, bailly@lille.inserm.fr
Received Decem ber 2, 2003
Accepted March 1, 2004
By a sligh tly m odified Han tzsch th iazole syn th esis eith er 1,3-bis[(th iazol-2-yl)am in o]ben zen e
derivatives 2 or 1,3-bis[2-im in oth iazol-3(2H)-yl]ben zen e derivatives 3 were exclusively ob-
tain ed. Th e com poun ds can be distin guish ed by NMR spectroscopy. Com poun ds 2a–2d an d
3
a–3d were evaluated for th eir poten tial an titum or activity, DNA in teraction , an d for th eir
activity again st DNA an d RNA viruses an d again st HIV-1 an d HIV-2.
Keyw ord s: Heterocycles; An tican cer agen ts; DNA-in teraction ; NMR spectroscopy; Th iazoles;
Han tzsch syn th esis.
Th e 2-am in oth iazole scaffold is presen t in several ph arm acoph ores. Deri-
1
vatives of 2-am in oth iazole h ave been studied as an tiin flam m atory , an ti-
2
3
bacterial an d an tiviral agen ts . Likewise, th is fun ction al group is presen t
4
in an tagon ists of fibrin ogen receptor sh owin g an tith rom bosis activity as
well as in n ew in h ibitors of bacterial DNA-gyrase B an d cyclin -depen den d
kin ases . Bleom ycin , belon gin g to th e fam ily of glycoprotein an tibiotics
5
6
an d con tain in g a bisth iazole structure, bin ds to DNA an d causes sin gle an d
7
double stran d breaks in DNA by a free radical m ech an ism . Bis-2-am in o-
th iazoles h ave been less studied, partly because th ey are less easily avail-
able. Th erefore, we developed a straigh tforward syn th etic procedure to
th ese com poun ds an d evaluated th eir activity in h um an leukem ia cell sys-
tem togeth er with th eir poten tial DNA bin din g an d an tiviral properties.
Th e com poun ds were obtain ed in a on e-step reaction from α-h alo keton es
with potassium th iocyan ate an d 1,3-ph en ylen ediam in e. Depen din g on
reaction con dition s eith er 1,3-bis[(th iazol-2-yl)am in o]ben zen e derivatives 2
Collect. Czech. Chem. Commun. (Vol. 69) (2004)
doi:10.1135/cccc20041491

1
492
Lagoja et al.:
or 1,3-bis[2-im in oth iazol-3(2H)-yl]ben zen e derivatives 3 were exclusively
obtain ed.
Chemistry
8
In a sligh tly m odified version of th e Han tzsch th iazole syn th esis , th e con -
den sation of 2 equivalen ts of α-h alo keton es with potassium th iocyan ate
(
2.5 equivalen ts) an d 1,3-ph en ylen ediam in e h ydroch loride (1 equivalen t)
in refluxin g m eth an ol yielded 1,3-bis[(th iazol-2-yl)am in o]ben zen e deriva-
tives 2 or 1,3-bis[2-im inothiazol-3(2H)-yl]benzene derivatives 3 (Schem e 1).
Startin g from th e th iocyan ato keton e 4 (path way a), upon th e reaction with
th e an ilin e derivative, th e im idoth iocarbam ate in term ediate 5 is form ed,
wh ich , after cyclization , yields th e correspon din g 1,3-bis[(th iazol-2-yl)-
am in o]ben zen e derivative 2. Th is reaction path way h as been previously
8
c
described for th e efficien t syn th esis of com poun ds of type 2 . Wh en th e
Sch iff base 6 is form ed first (path way b), n ucleoph ilic exch an ge of th e h alo
atom for th e th iocyan ate m oiety gives rise to an arylim in o th iocyan ate 7,
wh ich cyclizes to th e correspon din g 1,3-bis[2-im in oth iazol-3(2H)-yl]-
ben zen e derivative 3.
R1
CH3
R1
CH3
N
S
R1
CH3
N
a)
b)
H
N
S
R1
H
N
N
N
HN
H
R1
O
Br
S
1
NH
H3C
S
CH3
+
KSCN
2
3
1
,3-p-phenylenediamine
R1
O
CH3
R1 CH3
R1 CH3
R1
CH3
H
H
H
H
S
NHR
O
SCN
N
R
Br
R
N
S
C
N
NH
5
4
6
7
R1
R1
2, 4, 5
3, 6, 7
a
2,4-Cl2C6H3
3-BrC6H4
CH3
a
b
c
d
4-FC6H4
4-ClC6H4
4-BrC6H4
CH3
b
c
d
CO2CH3
Pathway a) 1. α-bromoketone, KSCN, MeOH, r.t. 1 h; 2. 1,3-phenylenediamine·HCl, reflux 18 h.
Pathway b) 1. α-bromoketone, 1,3-phenylenediamine·HCl, NaHCO3, MeOH, reflux 3 h; 2. KSCN,
reflux 18 h
SCHEME 1
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

1
,3-Dithiazolylbenzene Derivatives
1493
Th e syn th esis can be carried out as a on e-pot procedure an d, depen din g
on th e ch osen reaction con dition s, eith er com poun ds 2 or 3 are obtain ed
in good yields. Th is is dem on strated by th e syn th esis of 2c an d 3d , start-
in g from th e sam e α-h alo keton e (3-ch lorobutan -2-on e). Th ese fin din gs are
9
in con trast to earlier fin din gs wh ere on ly th e form ation of im in oth iazole
derivatives sim ilar to com poun ds 3, is claim ed. Th e reaction is suitable for
aliph atic as well as arom atic α-h alo keton es. Th e selection of α-brom o
keton es 1 th at were used in th e reaction was based both on availability an d
on attem pts to obtain a con siderable diversity of th e substitution pattern .
All com poun ds of series 2 an d 3 were ch aracterized by NMR spectroscopy;
all proton an d carbon reson an ces could be assign ed by m ean s of gHMQC
1
0
an d gHMBC spectroscopy . In spection of ch em ical sh ifts an d spectral lin e
sh apes of com poun ds 2 an d 3 sh owed several ch aracteristics wh ich distin -
guish both fam ilies of com poun ds. Am in oth iazoles 2 sh owed sh arp,
am in o-proton lin e sh apes an d rath er large ch em ical sh ifts (δ ≈ 10.1 ppm )
wh ereas th e im in o proton s of im in oth iazole com poun ds 3 produced very
broad sign als at lower ch em ical sh ifts (δ ≈ 9.7 ppm ).
Large ch em ical sh ift differen ces between com poun ds 2 (δ 135.5, 129.4,
1
31.8 an d 132.7 ppm – C-1/3, C-2, C-4/6 an d C-5, respectively) an d 3
(
δ 141.8, 104.7, 110.2 an d 129.6 ppm – C-1/3, C-2, C-4/6 an d C-5, respec-
tively) could be observed in th e cen tral arom atic rin g. Th ese differen ces are
due to better electron don or properties of th e am in o group in series 2 th an
th ose of th e im in o fun ction al group in series 3. Th e two isom ers can also be
easily distin guish ed by com parin g th e sign al correspon din g to carbon 2 of
th e th iazole m oiety. For am in oth iazoles 2, it was foun d in th e δ 160 ppm
region , wh ereas th e correspon din g sign al of th e 2-im in oth iazoles 3 was
foun d in th e δ 169 ppm region . Overlap of th e reson an ce sign als of th e
th iazole m oieties an d th eir substituen ts sh ows th ese m olecules to be sym -
m etrical. An en ergy m in im ized drawin g of com poun ds 2 an d 3 is sh own in
Fig. 1. Togeth er with NMR spectra an d h igh resolution m ass spectrom etry,
th e sh arp m eltin g poin ts are con firm in g th e h igh purity of th e com poun ds.
Biological Results and Discussion
Com poun ds 2a–2d an d 3a–3c were evaluated for th eir poten tial an titum or
activity. Th e drug con cen tration required to in h ibit th e growth of CEM h u-
m an leukem ia cells by 50% (IC ) was determ in ed an d th e data are listed in
5
0
Table I.
In th e 1,3-bis[(th iazol-2-yl)am in o]ben zen e series, 2a gave IC50 values in
th e low m icrom olar ran ge, wh ereas 2c an d 2d were of lower activity. In
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

1
494
Lagoja et al.:
th e 1,3-bis[2-im in oth iazol-3(2H)-yl]ben zen e series, m arked cytotoxic effects
were observed with all con gen ers; th e brom o (3c) an d ch loro (3b) deriva-
tives are som ewh at m ore active th an th e fluoro derivative (3a).
DNA in teraction was also evaluated by m eltin g tem perature (T ) an d
m
DNase I footprin tin g but little or n o in teraction was observed. DNA is ap-
paren tly n ot a direct target for th ese m olecules. However, cytom etry m ea-
surem en ts usin g brom odeoxyuridin e-labeled cells in dicated th at th e m ost
cytotoxic com poun ds 3b an d 3c in h ibit DNA syn th esis durin g th e S ph ase
of th e cell cycle (Fig. 2a). Moreover, up to 75% of CEM cells treated with
th e sam e com poun ds were positively stain ed with an n exin V (Fig. 2b) an d
in th e cell cycle m easurem en ts (propidium iodide stain in g), appearan ce of a
TABLE I
Con cen tration required in h ibitin g th e growth of CEM h um an leukem ia cells by 50% (IC50
)
Com poun d
R1
IC50, µM
2
2
2
2
3
3
3
a
b
c
d
a
b
c
2,4-Cl C H
2.53
>60
2
6
3
3-BrC H
6
4
CH3
23.66
12.85
5.22
CO CH
2
3
4-FC H
6
4
4-ClC H
1.63
6
4
4
4-BrC H
1.43
6
a
b
FIG. 1
1
1
1
3
En ergy m in im ized drawin g of a N ,N -bis[4-(3-brom oph en yl)-5-m eth yl-1,3-th iazol-2-yl]-
ben zen e-1,3-diam in e (2b) an d b 1,3-bis[4-(4-brom oph en yl)-2-im in oth iazol-5-m eth yl-3(2H)-yl]-
ben zen e (3c)
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1
,3-Dithiazolylbenzene Derivatives
1495
sub-G1 cell population was n oted, in dicatin g in duction of apoptosis by
th ese m olecules. Com poun ds 2a–2d an d 3a–3c h ave also been evaluated
for th eir activity again st DNA an d RNA viruses as well as again st HIV-1 an d
HIV-2. No specific an tiviral activity was observed.
control
Compound 3b
Compound 3c
4
3
2
1
1
1
0
0
0
a
4
6%
23%
20%
BrdU
1
0
1
0
1
0
0
1223
PI
4
1
0
0
0
b
3
.4%
33.9%
34.3%
3
2
1
1
PI
93.3%
36.2%
37.3%
1
0
1
1
0
0
3
.3%
29.4%
28.0%
0
1
2
3
4
1
0
10
10
10
10
annexin V
FIG. 2
Cytom etry experim en ts usin g brom odeoxyuridin e as m arker for DNA syn th esis (a) an d
an n exin V for apotosis (b)
Conclusion
Usin g a m odified Han tzsch syn th esis, dith iazolyl ben zen e derivatives are
available in good yields. Con trol of th e reaction con dition s allows directin g
th e reaction eith er to th e form ation of 1,3-bis[(th iazol-2-yl)am in o]ben zen e
derivatives 2 or 1,3-bis[2-im in oth iazol-3(2H)-yl]ben zen e derivatives 3.
Th ese dith iazolylben zen e derivatives are a n ovel series of poten tial an ti-
tum or agen ts; furth er ph arm acological studies are on goin g to elucidate
th eir m ech an ism of action .
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1
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Lagoja et al.:
EXPERIMENTAL
1
NMR spectra (δ, ppm ; J, Hz) were recorded on a Varian , Gem in i 200 spectrom eter ( H at 200
1
3
an d C at 50 MHz). All NH proton s were assign ed by exch an ge with D O. In AA′BB′ sys-
2
1
2
tem s, determ in ation of J is based on th e assum ption of an AB quartet . gHMQC an d
gHMBC NMR spectra were recorded on a Varian 500 MHz Un ity spectrom eter, operatin g at
4
99.505 MHz. Quadrature detection was ach ieved by States–Haberkorn h ypercom plex
1
3
m ode . Th e gHMQC an d gHMBC spectra in DMSO con sisted of 1024 data poin ts in t2 an d
12 in crem en ts in t . Th e data were apodized with a sh ifted sin e-bell square fun ction in
5
1
both dim en sion s an d processed to a 2K × 2K m atrix. Exact m ass m easurem en ts were per-
form ed on a quadrupole–tim e of fligh t m ass spectrom eter (Q-Tof-2, Microm ass, Man ch ester,
U.K.) equipped with a stan dard electrospray ion ization (ESI) in terface. Sam ples were in fused
in a propan -2-ol–water (1:1) m ixture at 3 µl/m in . TLC was perform ed with TLC alum in um
sh eets (Silica gel 60 F254, Merck) an d silica (200–425 m esh ) was used for colum n ch rom ato-
graph y. Meltin g poin ts were determ in ed with a Kofler block. All startin g m aterials are com -
1
4
m ercially available (e.g. from ACROS, Aldrich , Fluka) or can be obtain ed as outlin ed furth er
.
N,N′-Bis(th iazol-2-yl)ben zen e-1,3-diam in es 2a–2d . Gen eral Procedure (Path a)
A solution of th e α-h alo keton e (4.69 m m ol) an d potassium th iocyan ate (684 m g, 7.03 m m ol)
in m eth an ol (20 m l) was stirred at room tem perature for 1 h . Subsequen tly, 1,3-ph en ylen e-
diam in e h ydroch loride (424 m g, 2.34 m m ol) was added an d th e reaction m ixture was
refluxed overn igh t. Progress of th e reaction was ch ecked by TLC (eth er–petroleum eth er,
3
:1). After coolin g to am bien t tem perature, th e precipitate was filtered off an d wash ed with
water. Th e crude products were dried an d crystallized from MeOH–water.
1
3
N ,N -Bis[4-(2,4-dichlorophenyl)-5-methyl-1,3-thiazol-2-yl]benzene-1,3-diamine (2a). Yield
+
8
0%. R (eth er–petroleum eth er, 3:1) 0.42. M.p. 240 °C (MeOH, water). HRMS [M+H] calcu-
F
1
lated: 590.9805; foun d: 590.9814. H NMR: 2.11 s, 6 H (CH ); 7.13 m , 3 H (H-4,5,6 cen tral
ben zen e); 7.81 s, 1 H (H-2 cen tral ben zen e); 7.49 m , 4 H (H-5,6 R ); 7.72 m , 2 H (H-3 R );
0.10 s, 2 H (ex, NH). 13C NMR: 11.4 (CH ); 118.9, 142.7, 160.1 (C-5, C-4, C-2 th iazole);
3
1
1
1
1
1
3
04.7, 110.2, 129.6, 141.8 (C-2, C-4,6, C-5, C-1,3 cen tral ben zen e rin g); 127.4, 129.4, 133.4,
1
33.5, 133.6, 133.9 (C-5, C-3, C-2, C-6, C-1, C-4 R ).
1
3
N ,N -Bis[4-(3-bromophenyl)-5-methyl-1,3-thiazol-2-yl]benzene-1,3-diamine (2b). Yield 76%.
RF (eth er–petroleum eth er, 3:1) 0.38. M.p. 246 °C (MeOH, water). HRMS [M+H] calculated:
10.9574; foun d: 610.9576. H NMR: 2.40 s, 6 H (CH ); 7.19 m , 3 H (H-4,5,6 cen tral ben -
3
zen e); 7.36 dd, 2 H (H-5 R ); 7.47 d, 2 H (H-6 R ); 7.63 d, 2 H (H-4 R ); 7.82 s, 2 H (H-2 R );
.05 s, 1 H (H-2 cen tral ben zen e); 10.13 s, 2 H (ex, NH). C NMR: 12.0 (CH ); 117.9, 143.7,
59.6 (C-5, C-4, C-2 th iazole); 105.0, 110.3, 129.9, 141.9 (C-2, C-4,6, C-5, C-1,3 cen tral ben -
zen e rin g); 121.8, 127.0, 129.5, 130.6, 130.7, 137.6 (C-3, C-6, C-1, C-2, C-5, C-4 R ).
N ,N -Bis(4,5-dimethyl-1,3-thiazol-2-yl)-1,3-benzenediamine (2c) . Yield 75%. R (eth er–
petroleum eth er, 3:1) 0.30. M.p. 178 °C (MeOH, water). HRMS [M+H] calculated: 331.1051;
foun d: 331.1046. H NMR: 2.12 s, 6 H (CH3 R ); 2.18 s, 6 H (CH ); 6.65–6.89 m , 3 H
H-4,5,6 cen tral ben zen e); 7.85 s, 1 H (H-2 cen tral ben zen e); 9.81 s, 2 H (ex, NH). C NMR:
+
1
6
1
1
1
1
1
3
8
1
3
1
1
3
8c
F
+
1
1
3
1
3
(
1
1
1
0.5 (CH ); 14.6 (CH3 R ); 113.5, 142.6, 159.2 (C-5, C-4, C-2 th iazole); 104.4, 109.4, 129.1,
3
41.9 (C-2, C-4,6, C-5, C-1,3 cen tral ben zen e rin g).
Methyl 2-(3-{[4-(methoxycarbonyl)-5-methyl-1,3-thiazol-2-yl]amino}anilino)-5-methyl-1,3-
thiazole-4-carboxylate (2d ). Yield 73%. RF (eth er–petroleum eth er, 3:1) 0.28. M.p. 210 °C
(
MeOH, water). HRMS [M+H]+ calculated: 419.0848; foun d: 419.0845. 1H NMR: 2.57 s, 6 H
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

1
,3-Dithiazolylbenzene Derivatives
1497
1
(
CH ); 3.78 s, 6 H (CH CO R ); 7.24 m , 3 H (H-4,5,6 cen tral ben zen e); 7.65 s, 1 H (H-2 cen -
3
3
2
1
3
1
tral ben zen e); 10.27 s, 2 H (ex, NH). C NMR: 12.2 (CH ); 51.6, 162.7 (CH , CO R ); 133.1,
3
3
2
1
38.6, 159.1 (C-5, C-4, C-2 th iazole); 105.3, 110.3, 129.8, 141.5 (C-2, C-4,6, C-5, C-1,3 cen -
tral ben zen e rin g).
1
,3-Bis[2-im in oth iazol-3-(2H)-yl]ben zen es 3a–3d . Gen eral Procedure (Path b)
A solution of th e α-h alo keton e (4.69 m m ol), 1,3-ph en ylen ediam in e h ydroch loride (424 m g,
2
.34 m m ol) an d NaHCO3 (394 m g, 4.69 m m ol) in m eth an ol (20 m l) was refluxed for 3 h .
After disappearan ce of th e startin g m aterial (TLC), potassium th iocyan ate (684 m g, 7.03 m m ol)
was added an d h eatin g un der reflux con tin ued overn igh t. Progress of th e reaction was
ch ecked by TLC (eth er–petroleum eth er, 3:1). After coolin g to am bien t tem perature, th e pre-
cipitate was filtered off an d wash ed with water. Th e crude products were dried an d crystal-
lized from MeOH–water.
1
,3-Bis[4-(4-fluorophenyl)-2-iminothiazol-5-methyl-3(2H)-yl]benzene (3a ). Yield 74%.
+
RF (eth er–petroleum eth er, 3:1) 0.13. M.p. 246 °C (MeOH, water). HRMS [M+H] calculated:
91.1175; foun d: 491.1176. 1H NMR: 2.10 s, 6 H (CH ); 7.12 m , 8 H (H-2,6,3,5 R ); 7.46 m ,
H (H-4,5,6 cen tral ben zen e); 7.99 s, 1 H (H-2 cen tral ben zen e); 9.60 s, 2 H (ex, NH).
C NMR: 11.8 (CH ); 116.6, 132.7, 168.0 (C-5, C-4, C-2 th iazole); 129.4, 131.8, 132.7, 135.5
1
4
3
3
1
3
3
(
C-2, C-4,6, C-5, C-1,3 cen tral ben zen e rin g); 116.0 (d), 133.5 (d), 135.4 (d), 165.1 (d) (C-3,5,
1
C-2,6, C-1, C-4 R ).
1
,3-Bis[4-(4-chlorophenyl)-2-iminothiazol-5-methyl-3(2H)-yl]benzene (3b ). Yield 73%.
+
RF (eth er–petroleum eth er, 3:1) 0.12. M.p. 252 °C (MeOH, water). HRMS [M+H] calculated:
23.0584; foun d: 523.0585. H NMR: 2.11 s, 6 H (CH ); 7.12 d, 4 H (H-3,5 R ); 7.39 d, 4 H
H-2,6 R ); 7.46 m , 3 H (H-4,5,6 cen tral ben zen e); 8.07 s, 1 H (H-2 cen tral ben zen e); 9.85 s,
H (ex, NH). C NMR: 11.3 (CH ); 116.6, 134.4, 167.9 (C-5, C-4, C-2 th iazole); 129.4,
31.7, 132.6, 135.5 (C-2, C-4,6, C-5, C-1,3 cen tral ben zen e rin g); 126.5, 128.9, 132.5, 134.8
C-4, C-3,5, C-2,6, C-1 R ).
1
1
5
(
2
1
(
3
1
1
3
3
1
1
,3-Bis[4-(4-bromophenyl)-2-iminothiazol-5-methyl-3(2H)-yl]benzene (3c). Yield 77%.
+
RF (eth er–petroleum eth er, 3:1) 0.09. M.p. 258 °C (MeOH, water). HRMS [M+H] calculated:
10.9574; foun d: 610.9577. H NMR: 2.11 s, 6 H (CH ); 7.05 d, 4 H (H-3,5 R ); 7.46 d, 4 H
H-2,6 R ); 7.48 m , 3 H (H-4,5,6 cen tral ben zen e); 8.10 s, 1 H (H-2 cen tral ben zen e); 9.78 s,
H (ex, NH). C NMR: 12.0 (CH ); 116.6, 132.7, 167.9 (C-5, C-4, C-2 th iazole); 129.4,
31.6, 132.7, 135.5 (C-2, C-4,6, C-5, C-1,3 cen tral ben zen e rin g); 123.7, 126.8, 131.9, 134.4
C-3,5, C-4, C-2,6, C-1 R ).
1
1
6
(
2
1
(
3
1
1
3
3
1
1
,3-Bis[4,5-dimethyl-2-iminothiazol-3(2H)-yl]benzene (3d ). Yield 70%. R (eth er–petroleum
F
+
eth er, 3:1) 0.09. M.p. 193 °C (MeOH, water). HRMS [M+H] calculated: 331.1051; foun d:
31.1049. 1H NMR: 2.14 s, 6 H (CH3 R ); 2.20 s, 6 H (CH ); 7.15 m , 2 H (H-4, 6 cen tral ben -
1
3
3
zen e); 7.40 m , 1 H (H-5 cen tral ben zen e); 7.90 s, 1 H (H-2 cen tral ben zen e); 10.0 s, 2 H (ex,
1
3
1
NH). C NMR: 10.5 (CH ); 13.5 (CH R ); 114.0, 131.2, 165.0 (C-5, C-4, C-2 th iazole);
3
3
1
29.3, 131.6, 131.8, 135.4 (C-3,5, C-4, C-2,6, C-1 cen tral ben zen e rin g).
Experim en tal procedures for th e cell cultures
1
5a
an d survival assay15b, cell cycle an aly-
sis15c, extern alization of ph osph atidylserin es an d brom odeoxyuridin e in corporation 15d h ave
been described previously.
The authors thank Dr J. Rozenski for Mass Analysis, Prof. E. De Clercq and Dr C. Pannecouque for
antiviral screening. K. Nauwelaerts is a research assistant of the FWO.
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

1
498
Lagoja et al.:
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