Synthesis and biological activities of new 1,2,4-triazol-3-one derivatives

Article abstract of DOI:10.1007/s11171-005-0054-0

New 2-phenacyl-1,2,4-triazol-3-ones were obtained by the reaction of 5-alkyl-1,2,4-triazol-3-ones with α-bromoacetophenone in an alkaline medium. Selective reduction of the side chain carbonyl group to hydroxy group was achieved with NaBH₄. The

Full text of DOI:10.1007/s11171-005-0054-0

Russian Journal of Bioorganic Chemistry, Vol. 31, No. 4, 2005, pp. 387–397. Translated from Bioorganicheskaya Khimiya, Vol. 31, No. 4, 2005, pp. 430–440.
Original Russian Text Copyright © 2005 by N. Demirba£, A. Demirba£, Karao≠lu.
Synthesis and Biological Activities
of New 1,2,4-Triazol-3-one Derivatives1
2
N. Demirba£, A. Demirba£, and ‹. A. Karao≠lu
Department of Chemistry, Karadeniz Technical University, Trabzon, 61080 Turkey
Received December 1, 2004; in final form, March 14, 2005
Abstract—New 2-phenacyl-1,2,4-triazol-3-ones were obtained by the reaction of 5-alkyl-1,2,4-triazol-3-ones
with α-bromoacetophenone in an alkaline medium. Selective reduction of the side chain carbonyl group to
hydroxy group was achieved with NaBH4. The reaction of some compounds containing a phenolic hydroxyl
with 4-toluenesulfonyl chloride or benzyl bromide in the presence of NaOH led to tosylated or benzylated
derivatives. The tosylation or benzylation at the alcoholic hydroxyl was carried out in the presence of sodium
metal. Some of the newly synthesized compounds revealed an antimicrobial activity; 6 of 14 new compounds
that were studied by the National Cancer Institute were found to possess antitumor activity.
Key words: antimicrobial activity, antitumor activity, benzylation, reduction, tosylation, 1,2,4-triazol-3-one
21
INTRODUCTION
1,2,4-triazol-3-one ring exhibit a selective antitumor
activity only towards breast cancer [8].
In the last decades, the drug resistance to the com-
Sulfonates, which have been used as important
intermediates in organic synthesis and possess antivi-
ral, anti-HIV or anticancer activity [15, 16], are usually
obtained by treatment of the corresponding alcohols
with sulfonating agents in the presence of pyridine or
triethylamine. Moreover, tetramethylalkylidenedi-
amine and silver(I) oxide were reported to be efficient
reagents for the sulfonylation of alcohols in the pres-
ence of sulfonyl chlorides; the latter reagent is being
used in the presence of a catalytic amount of potassium
iodide [17]. In addition, several sulfonylation reactions
were achieved in aqueous sodium hydroxide when suf-
ficiently hydrophilic alcohols were used [18, 19].
monly used antibiotics has become a wide-spread and
serious problem. Therefore, the synthesis of new com-
pounds that could be used for the effective treatment of
infectious diseases without side effects is an urgent bio-
medical problem.
1,2,4-Triazoles are a new class of synthetic thera-
peutic agents. For instance, fluconazole and itracona-
zole are used as antimicrobial drugs in medicine [1, 2].
Moreover, vorozole, letrozole and anastrozole are non-
steroidal drugs used for the treatment of cancer [3]. In
addition, a number of triazole derivatives have been
reported to possess pharmacological, insecticidal, fun-
gicidal and herbicidal activities [4–11]. Some N-alky-
lated derivatives of 1,2,4-triazoles were obtained in our
laboratory as antimicrobial agents by the reaction of
5-alkyl-1,2,4-triazol-3-one with the corresponding
alkyl halide in the presence of sodium ethoxide [5, 6].
The S-alkylation of 1,2,4-triazol-3-thiones was
achieved in aqueous sodium hydroxide in the case of
the precursors that have a good solubility in water [12].
Furthermore, some Schiff base derivatives of 1,2,4-tri-
azol-3-ones and their reduced derivatives were
obtained as antitumor agents in our laboratory [7, 13,
14]. The highest activity was observed for the com-
pounds containing a phenylethylidenamino- or phenyl-
ethylamino group in the position 4 of the 1,2,4-triazol-
3-one ring. On the other hand, the compounds contain-
ing an arylidene-hydrazide group in position 2 of the
These facts allow us to hope for the developing bio-
active compounds and, in this study, to synthesize a
series of new 1,2,4-triazol-3-one derivatives.
RESULTS AND DISCUSSION
We first synthesized ten new 1,2,4-triazol-3-one
derivatives (IIa)–(IIj) by the reaction of (Ia)–(Ij) with
α-bromoacetophenone in ethanolic sodium ethoxide
(Scheme 1). The structures of new compounds were
confirmed by IR, 1H and 13C NMR and mass (for some
selected compounds) spectra. The spectral data were
consistent with the proposed structures. In the NMR
spectra of (IIa)–(IIj), additional signals from acetophe-
none moiety were observed, while the NH signal was
absent. The IR spectra of these compounds display an
additional absorption peak belonging to exocyclic car-
bonyl function and no signals belonging to NH group.
In addition, (IIa), (IIc), and (IIg) gave good molecular
ion peaks.
1
The text was submitted by the authors in English.
Corresponding author; phone: +90 (462) 377-2600; fax:+90-
(462)-325-31-95; e-mail: neslihan@ktu.edu.tr.
2
1068-1620/05/3104-0387 © 2005 Pleiades Publishing, Inc.

388
DEMIRBA‹ et al.
N
N
N
N
F
N
Cl
Cl
N
N
N
HO
N
O
N
CH3
CH3
O
O
H
N
N
N
F
O
N
N
Fluconazol
Itraconazol
N
N
N
N
N
N
CH3
N
N
N
NC
H3C
N
N
N
H3C
Cl
NC
Cl
H3C
CH3
CN
Anastrozol
Vorozol
Letrozol
Compounds (IIIa)–(IIIj) were obtained by the while additional new signals due to a tosyl group with
1
selective reduction by NaBH4 of exocyclic carbonyl
the corresponding chemical shifts appear. In the H
group in (IIa)–(IIj) and imine bond in (IIc)–(IIj) NMR spectrum of (VI), the resonance assigned to alco-
(Scheme 1). No reduction of heterocyclic ring took holic OH is observed at 5.98 ppm. On the other hand,
place in the course of these reactions since NaBH4 was the resonance of alcoholic OH of starting (IIIb) disap-
pears from the 1H NMR spectrum of (VII). In addition,
(IV), (V) and (VI) gave molecular ion peaks consistent
with their structures.
used as the reducing agent [20]. Note that the reduction
of (IIa)–(IIj) was achieved in milder conditions then
similar reactions carried out in our laboratory [7, 13,
14]. The spectra of (IIIa)–(IIIj) are very informative.
In the IR spectra of (IIIa)–(IIIj), only one carbonyl
absorption band is observed as a result of selective
reduction of exocyclic carbonyl in (IIa)–(IIj). In the 1H
NMR spectra of reduced compounds (IIIa)–(IIIj), the
CHOH group resonates at 4.9–5.4 ppm (1 H) and 5.5–
5.7 ppm (1 H), NCH2 group exhibits doublets centered
at 3.6–4.3 and 3.8–4.5 ppm (2 H), NHCH2 proton reso-
nates at 3.7–4.2 ppm for (IIIc)–(IIIg) and 6.2–6.3 ppm
for (IIIh)–(IIIj).
In the 13C NMR spectra of these compounds, the
CH2–NH carbons of Y substituent resonate at 47–
49 ppm for (IIIc)–(IIIj), carbons of CH–OH group, at
69–71 ppm, while the N=CH and exocyclic C=O car-
bon signals were absent.
The derivatives (IIg) and (IIIg) were benzylated by
the treatment with benzyl bromide in ethanol in the
presence of sodium hydroxide. The NMR spectra of the
resulting (VIII) and (IX) exhibit no signals from phe-
nolic hydroxyl; new signals assigned to benzyl group
appear instead. On the other hand, the benzylation of
(IIIa) and (IIId) in the presence of sodium metal in dry
toluene led to (X) and (XI), respectively (Scheme 3).
As a result, the signals of alcoholic hydroxyls are
absent from the IR and 1H NMR spectra of these com-
pounds; instead, additional signals are observed due to
benzyl group at the corresponding chemical shifts. On
the other hand, the signal assigned to alcoholic OH is
present in the 1H NMR spectrum of (IX), while the res-
onance from phenolic OH is absent.
Tosylated compounds (IV), (V) and (VI) were syn-
thesised by the reaction of the corresponding (IIg),
(IIj), and (IIIj) with 4-toluenesulfonyl chloride in eth-
anol in the presence of NaOH; no acylation reaction
took place under these conditions at alcoholic OH of
(IIIj). On the other hand, the tosylation of (IIIb) could
be achieved in dry toluene in the presence of sodium
metal (Scheme 2). The IR spectra of (IV) and (V) dis-
play no absorption bands derived from aromatic OH
group. The proton signal belonging to aromatic OH is
Among the compounds tested, (IIc), (IIh), and
(IIIj) exhibit good activities against Staphylococcus
aureus and Bacillus subtilis with the MIC values of
1 µg/ml. Moreover, (IIc) exhibits a good activity
against Enterococcus faecalis, whereas (IIj) is active
against B. subtilis and moderately active against E.
faecalis and S. aureus with the MIC values of 1, 9, and
4 µg/ml, respectively. An increased activity toward S.
aureus was observed with the MIC value of 1 µg/ml,
when (IIj) was converted to (IIIj). The tosylation of
absent from the 1H NMR spectra of these compounds, phenolic hydroxyl in (IIj) highly increased the antimi-
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 4 2005

SYNTHESIS AND BIOLOGICAL ACTIVITIES
O
389
OH
O
Br
N
NH
N
N
N
N
NaBH4
R
N
X
O
R
N
X
O
R
N
Y
O
(Ia)–(Ij)
(IIa)–(IIj)
(IIIa)–(IIIj)
(I)–(III)
R
X
Y
a
b
c
d
e
f
CH3
NH2
NH2
NH2
NH2
CH2C6H5
CH3
N=CHC6H5
N=CHC6H5
NHCH2C6H5
C6H5
NHCH2C6H5
CH2C6H5
N=CHC6H3Cl(2,4)
N=CHC6H3Cl(2,4)
N=CHC6H4OH(o-)
N=CHC6H4OH(o-)
N=CHC6H4OH(o-)
N=CHC6H4OH(o-)
NHCH2C6H3Cl(2,4)
NHCH2C6H3Cl(2,4)
NHCH2C6H4OH(o-)
NHCH2C6H4OH(o-)
NHCH2C6H4OH(o-)
NHCH2C6H4OH(o-)
CH2C6H4Cl(-p)
CH3
g
h
i
C2H5
C3H7
j
CH2C6H5
Scheme 1.
crobial effect of the resulting (V) toward E. faecalis, E. faecalis, S. aureus, and B. subtilis with the MIC val-
while decreasing the activity against B. subtilis.
ues of 4 µg/ml.
Similarly, tosylation of (IIIj) increased the activity
against E. faecalis and decreased the observed effects
for S. aureus and B. subtilis. Compound (IIb) mani-
fested a good activity against Escherichia coli (MIC
value of 2 µg/ml) and moderate activities for E. faeca-
lis, S. aureus, and B. subtilis (MIC values = 8, 4 and
4 µg/ml). The reduction of (IIb) to (IIIb) decreased the
activity against E. coli and S. aureus up to MIC values
of 8 µg/ml. Decreased activities toward E. coli,
E. faecalis, S. aureus, and B. subtilis (up to MIC values
of 16, 16, 32 and 16 µg/ml, respectively) were observed
for (VII), the derivative of (IIIb) tosylated at alcoholic
OH. On the other hand, the effect of tosyl group at phe-
nolic hydroxyl is more complex, since it increases the
activity against E. faecalis, while decreasing the anti-
microbial effect toward S. aureus and B. subtilis. In
addition, (IV), the tosylated derivative of (IIg),
demonstrates decreasing activities against E. faecalis,
S. aureus, and B. subtilis in comparison with the parent
compound. A similarly decreased activity in the same
tests was observed for (IIIc), a reduced derivative of
(IIc). However, the reduction of (IIh) to (IIIh)
increases the activity against E. faecalis, while decreas-
ing the activity against S. aureus and B. subtilis. Mod-
The benzylation of (IIg), (IIIc) and (IIIg) led to a
decrease in antimicrobial activity. On the contrary, the
benzylation of (IIId) at alcoholic hydroxyl led an
increase in the activity against E. faecalis, S. aureus,
and B. subtilis (MIC values of 0.3 µg/ml). All the tested
compounds exhibited no activity against Candida albi-
cans, C. tropicalis, and C. glabrata. DMSO, acetone,
and ethanol showed no inhibitory effect on Gram-posi-
tive and Gram-negative bacteria.
Among the compounds with a tosyl group in their
molecules (IV)–(VII), the reduced derivatives (VI) and
(VII) were found to possess antitumor activity, whereas
(IV) and (V) that simultaneously contain a tosyl group
and an Schiff base exhibited no antitumor activity.
Among the benzylated compounds (VIII)–(XI), only
(IX), reduced and benzylated at phenolic hydroxyl, dis-
played an antitumor activity. These results do not per-
mit any evaluation of structure–antitumor activity rela-
tionship within the series of (IIa)–(IIj) and (IIIa)–
(IIIj), since (IIi) displays an antitumor activity, while
its reduced derivative (IIIi) not. On the contrary, (IIf)
was found to be inactive, while its reduced derivative
erate activities were observed for (IIa) and (IIf) against (IIIf) do possess an antitumor activity.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 4 2005

390
DEMIRBA‹ et al.
(IIg)
(IIj)
(IIIj)
TsCl
TsCl
TsCl
O
O
OH
N
N
N
N
N
N
H3C
N
N
O
N
O
N
O
HN
HN
O
O
O
O S O
O S O
O S O
CH3
CH3
CH3
(IV)
(V)
(VI)
H3C
O S O
O
TsCl
(IIIb)
N
N
N
O
NH2
(VII)
Scheme 2.
EXPERIMENTAL
amino(or
arylidenamino)-2,4-dihydro-3H-1,2,4-tri-
azol-3-one (I) (0.01 mol) was refluxed with equivalent
amount of sodium metal in absolute ethanol for 2 h.
Then, α-bromoacetophenone (0.01 mol) was added,
and the mixture was refluxed for additional 5 h. After
evaporation at 35–40°ë in a vacuum, the resulting solid
was recrystallized from an appropriate solvent.
Melting points were determined on a Gallenkamp
melting point apparatus and are uncorrected. 1H NMR
and 13C NMR spectra were recorded on a Varian-Mer-
cury 200 MHz spectrometer in DMSO-d6, chemical
shift values are given in ppm (δ scale), and tetramethyl-
silane was used as an internal standard. IR spectra were
recorded in potassium bromide pellets on a Perkin-
Elmer 1600 FTIR spectrometer (ν, cm–1). Mass spectra
were recorded on a Quattro LC-MS (70 eV) Instru-
ment. All the chemicals were obtained from Fluka Che-
mie AG Buchs (Switzerland). Compounds (Ia)–(Ij)
were synthesized by the earlier published method [23].
4-Amino-5-methyl-2-phenacyl-2,4-dihydro-3H-
1,2,4-triazol-3-one (IIa); yield 82.20%, mp 182–
183°ë (from ethanol); IR: 1723 and 1699 (2 C=O),
1611 (C=N) cm–1; H NMR: 1.85 (3 H, s, CH3), 5.43
1
(2 H, s, NCH2), 5.52 (2 H, s, NH2), 7.20–7.31 (3 H, m,
Ar), 7.43–7.58 (2 H, m, Ar); C NMR: 22.34 (CH3),
13
56.66 (NCH2), Ar: 128.01 (CH), 128.36 (2 CH), 129.61
(CH), 130.73 (CH), 131.24 (C), 143.04 (C3 of triazole),
154.18 (C5 of triazole), 194.21 (CH2C=O); MS, m/z
A general procedure for the synthesis of 5-alkyl-
4-X-2-phenacyl-2,4-dihydro-3H-1,2,4-triazol-3-ones
(IIa–IIj). The corresponding 5-alkyl(or aryl)-4-
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 4 2005

SYNTHESIS AND BIOLOGICAL ACTIVITIES
391
O
OH
N
N
N
N
H3C
N
N
O
H3C
N
O
HN
O
O
i
i
(IIg)
(IIIg)
(IIIa)
(IIId)
(VIII)
(IX)
i
i
O
O
N
N
O
N
N
N
H3C
N
O
HN
NH2
(XI)
(X)
i – BrCH2C6H5.
Scheme 3.
(I, %): 238.08 [M + 1]+ (16), 233.08 [M]+ (100), 127.08 8.05–8.09 (2 H, m, Ar), 9.72 (H, s, N=CH); 13C NMR:
(33), 119.11 (23), 115.15 (25).
10.93 (CH3), 51.62 (NCH2), Ar: 127.58 (CH), 127.74
(2 CH), 128.08 (2 CH), 128.87 (2 CH), 128.93 (2 CH),
and 131.55 (CH), 133.18 (2C), 143.63 (C3 of triazol),
150.12 (C5 of triazol), 154.16 (N=CH), 192.84
(CH2C=O); MS, m/z (I, %): 321.04 [M]+ (100), 248.99
(50), 238.77 (14), 123.78 (14), 121.91 (27), 115.89
(12), 65.00 (36), 63.12 (60), 60.24 (18).
4-Amino-5-benzyl-2-phenacyl-2,4-dihydro-3H-
1,2,4-triazol-3-one (IIb); yield 80.12%, mp 188–
189°ë (from 1 : 1 ethyl acetate–petroleum ether); IR:
3308 and 3206 (NH2), 1706 and 1682 (2 C=O), 1652
(C=N) cm–1; 1H NMR: 3.85 (2 I, s, CH2), 5.38 (2 H, s,
NCH2), 5.42 (2 H, s, NH2), 7.28–7.36 (3 H, m, Ar),
7.58–7.72 (2 H, m, Ar), 7.69–7.98 (5 H, m, Ar); 13C
NMR: 37.87 (CH2), 51.66 (NCH2), Ar: 128.01 (2 CH),
128.12 (2 CH), 128.69 (2 CH), 129.32 (CH), 129.67
(CH), 130.12 (CH), 130.45 (CH), 131.19 (2 C)], 142.52
(C3 of triazol), 152.34 (C5 of triazol), 193.71
(CH2C=O).
5-Phenyl-4-(phenylmethyliden)amino-2-phenacyl-
2,4-dihydro-3H-1,2,4-triazol-3-one (IId); yield 81.62%;
mp 159–160°ë (from ethanol); IR: 1701 and 1692
(2 C=O), 1597 and 1544 (2 C=N) cm–1; 1H NMR: 5.61
(2 H, s, NCH2), 7.56–7.74 (6 H, m, Ar), 7.75–7.84 (3 H,
m, Ar), 7.87–7.93 (3 H, m, Ar), 8.05–8.12 (3 H, m, Ar),
9.67 (H, s, N=CH); 13C NMR: 54.13 (NCH2), Ar:
5-Methyl-4-(phenylmethyliden)amino-2-phena-
cyl-2,4-dihydro-3H-1,2,4-triazol-3-one (IIc); yield 125.89 (C), 127.88 (3 CH), 127.92 (3 CH), 128.11
74.13%); mp 136–137°ë (from ethanol); IR: 1703 and
1691 (2 C=O), 1611 and 1595 (2 C=N) cm–1; 1H NMR:
2.35 (3 H, s, CH3), 5.41 (2 H, s, NCH2), 7.54–7.59 (4 H,
m, Ar), 7.62–7.68 (2 H, m, Ar), 7.85–8.00 (2 H, m, Ar),
(2 CH), 128.56 (2 CH), 128.86 (CH), 129.02 (CH),
130.31 (CH), 131.77 (CH), 132.96 (CH), 134.00 (C),
and 134.11 (C), 143.68 (C3 of triazol), 150.22 (C5 of
triazol), 157.09 (N=CH), 192.64 (CH2C=O).
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 4 2005

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DEMIRBA‹ et al.
Table 1. Antimicrobial activity of the compounds tested (µg/ml)
Compound
(IIa)
(IIb)
(IIc)
(IIf)
(IIg)
(IIh)
(IIj)
(IIIb)
(IIIc)
(IIId)
(IIIh)
(IIIj)
(IV)
(V)
(VI)
(VII)
(VIII)
(IX)
(X)
(XI)
Amp.
Flu.
Ec
Kp
39
Yp
Pa
Ef
Sa
Bs
Ca
Ct
Cg
>1250
2
156
3125
156
156
3125
>1250
156
4
8
2
4
9
9
9
8
4
4
1
4
4
2
4
8
9
4
4
1
4
4
2
1
4
4
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
48
78
625
312
312
312
312
8
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
156
156
156
156
781
156
156
156
156
48
156
156
156
>1250
156
312
625
3125
312
3125
312
312
625
312
312
16
19
625
>1250
>1250
312
19
4
19
19
156
4
1
4
1
156
39
32
3125
6250
3125
3125
3125
3125
3125
94
3125
3125
3125
3125
3125
3125
3125
>1250
>128
32
4
16
4
16
3125
48
0.5
8
8
16
32
32
32
4
4
16
97
32
32
32
32
16
16
32
32
16
48
16
48
32
48
>1250
8
12
0.3
0.3
0.3
<1
32
32
2
2
32
<1
8
64
Note: Abbreviations: Ec, E. coli ATCC 25922; Kp, K. pneumoniae ATCC 13883; Yp, Y. pseudotuberculosis ATCC 911; Pa, P. aeruginosa ATCC
10145; Ef, E. faecalis ATCC 29212; Sa, S. aureus ATCC 25923; Bs, B. subtilis ATCC 6633; Ca, C. albicans ATCC 60193; Ct, C. tropi-
calis ATCC 13803; Cg, C. glabrata ATCC 66032. Amp., Ampicillin; Flu., Fluconazole; and (–), not determined (5 mg/ml).
5-Benzyl-4-[(2,4-dichlorophenyl)methyliden]ami- (C), 134.07 (2 CH), 134.19 (C), 135.07 (C), 136.73 (C),
no-2-phenacyl-2,4-dihydro-3H-1,2,4-triazol-3-one
(IIe); yield 84.21%; mp 167–168°ë (from ethanol); IR:
1709 and 1697 (2 C=O), 1595 and 1583 (2 C=N) cm–1;
1H NMR: 4.10 (2 I, s, CH2), 5.47 (2 H, s, NCH2), 7.27–
7.29 (5 H, m, Ar), 7.58–7.61 (3 H, m, Ar), 7.69–7.72 (2
H, m, Ar), 7.98–8.06 (3 H, m, Ar), 10.09 (H, s, N=CH);
13C NMR: 36.64 (CH2), 58.76 (NCH2), Ar: 126.74
(CH), 128.05 (3 CH), 128.43 (3 CH), 128.68 (3 CH),
128.83 (3 CH), 129.54 (C), 133.96 (C), 134.06 (C),
135.04 (C), 136.69 (C), 144.42 (C3 of triazol), 147.95
(N=CH), 150.02 (C5 of triazol), 192.69 (CH2C=O).
145.15 (C3 of triazol), 148.10 (N=CH), 150.00 (C5 of
triazol), 192.56 (CH2C=O).
4-[(2-Hydroxyphenyl)methyliden]amino-5-methyl-
2-phenacyl-2,4-dihydro-3H-1,2,4-triazol-3-one (IIg);
yield 81.38%; mp 175–176°ë (from 1 : 1 ethyl acetate–
petroleum ether); IR: 3327 (OH), 1724 and 1692 (2
1
C=O), 1614 and 1590 (2 C=N) cm–1; H NMR: 2.37
(3 H, s, CH3), 5.48 (2 H, s, NCH2), 7.14–7.31 (2 H, m,
Ar), 7.33–7.38 (2 H, m, Ar), 7.45–7.62 (3 H, m, Ar),
7.70–7.95 (2 H, m, Ar), 9.83 (H, s, N=CH), 10.29 (H, s,
OH); 13C NMR: 19.74 (CH3), 53.06 (NCH2), Ar:
118.82 (CH), 119.29 (2 CH), 119.57 (CH), 124.86
(CH),126.17 (CH), 128.21 (CH),129.48 (2 CH), 132.11
(CH), 134.05 (C), and 142.11 (C), 146.92 (C3 of tria-
zol), 155.37 (C5 of triazol), 162.63 (N=CH), 191.49
(CH2C=O); MS, m/z (I, %): 337.09 [M]+ (67), 304.17
(46), 282.17(44), 254.96 (21), 232.82 (14), 218.84 (13),
196.90 (11), 176.84 (16), 160.66 (11), 158.78 (59),
155.77 (100), 152.95 (51), 139.78 (16), 121.79 (13),
4-[(2,4-Dichlorophenyl)methyliden]amino-5-(4-
chlorophenyl)methyl-2-phenacyl-2,4-dihydro-3H-
1,2,4-triazol-3-one (IIf); yield 77.35%; mp 186–
187°ë (from ethanol); IR: 1705 and 1698 (2 C=O),
1
1587 and 1563 (2 C=N) cm–1; H NMR: 4.18 (2 H, s,
CH2), 5.46 (2 H, s, NCH2), 7.39 (4 H, br.s., Ar), 7.58–
7.61 (5 H, m, Ar), 7.80–8.03 (3 H, m, Ar), 9.89 (H, s,
N=CH); 13C NMR: 29.98 (CH2), 51.78 (NCH2), Ar:
128.05 (3 CH), 128.37 (2 CH), 128.84 (2 CH), 129.55
(CH), 129.59 (C), 130.66 (2 CH), 131.46 (C), 133.94 119.78 (34).
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393
Table 2. Antitumor activity of some compounds
4-[(2-Hydroxyphenyl)methyliden]amino-5-ethyl-
2-phenacyl-2,4-dihydro-3H-1,2,4-triazol-3-one (IIh);
yield 68.66%, mp 155–157°ë (from ethanol); IR: 3355
(OH), 1712 and 1693 (2 C=O), 1602 and 1591 (2 C=N)
cm–1; 1H NMR: 1.22 (3 H, t, J 7.0 Hz, CH3), 2.50 (2 H,
q, J 7.0 Hz, CH2), 5.41 (2 H, s, NCH2), 6.93–6.98 (2H,
m, Ar), 7.36–7.54 (H, m, Ar), 7.68–7.72 (4 H, m, Ar),
8.02–8.07 (2 H, m, Ar), 9.94 (H, s, N=CH), 10.40 (H, s,
OH); 13C NMR: 19.62 (CH2), 28.55 (CH2), 52.03
(NCH2), Ar: 116.11 (CH), 118.82 (CH), 119.43 (C),
124.78 (CH), 127.14 (CH), 128.82 (CH), 132.12 (CH),
133.28 (2 CH), 134.06 (2 C), 145.92 (C), 147.11 (C3 of
triazol), 156.74 (C5 of triazol), 162.69 (N=CH), 192.67
(CH2C=O).
Growth percentage
Number
Com-
pound
of tumor cell, %
assigned
by NCI
Activity
MCF7 NCI-H460 SF-268
(IIa)
(IIb)
(IIg)
(IIf)
729340
729341
731549
729346
729348
729343
729347
729901
734565
734567
734566
734568
734568
734564
122
40
46
62
29
1
102
96
93
96
94
32
4
99
90
–
–
–
–
+
+
+
–
–
–
+
+
–
+
121
96
(IIi)
80
(IIIb)
(IIIf)
(IIIi)
(IV)
(V)
(VI)
(VII)
(VIII)
(IX)
23
1
3
138
97
95
7
156
104
86
48
6
96
4-[(2-Hydroxyphenyl)methyliden]amino-5-pro-
pyl-2-phenacyl-2,4-dihydro-3H-1,2,4-triazol-3-one
(IIi); yield 66.52%, mp 156–157°ë (from ethanol); IR:
3355 (OH), 1708 and 1685 (2 C=O), 1601 and 1584 (2
C=N) cm–1; 1H NMR: 0.96 (3 H, t, J 7.0 Hz, CH3), 1.34
(2 H, m, CH2), 2.57–2.64 (2 H, d, J 6.0 Hz, CH2), 5.42
(2 H, s, NCH2), 6.93–6.97 (2 H, m, Ar), 7.33–7.43 (H,
m, Ar), 7.55–7.85 (4 H, m, Ar), 8.03–8.07 (2 H, m, Ar),
9.95 (H, s, N=CH), 10.35 (H, s, OH); 13C NMR: 13.28
(CH3), 19.62 (CH2), 28.52 (CH2), 51.68 (NCH2), Ar:
116.32 (CH), 119.33 (CH), 119.43 (C), 126.00 (CH),
128.03 (CH), 128.83 (CH), 130.71 (2 CH), 133.13
(CH), 134.05 (2 C), 146.07 (C), 147.26 (C3 of triazol),
157.51 (C5 of triazol), 162.69 (N=CH), 192.84
(CH2C=O).
106
89
85
0
53
110
1
112
30
118
45
NH2), 1688 (C=O), 1614 and 1519 (2 C=N) cm–1; H
NMR: 1.93 (3 H, s, CH3), 3.90 (H, d, NCH2), 4.07 (H,
d, NCH2), 5.43 (H, br. s, CH), 5.56 (H, d, J 4.0 Hz, OH),
6.52 (H, s, NH), 7.03–7.22 (3 H, m, Ar), 7.28–7.42
1
13
(2 H, m, Ar); C NMR: 23.65 (CH3), 57.61 (NCH2),
69.76 (CH), Ar: 126.11 (CH), 128.36 (CH), 129.19
(CH), 128.91 (CH), 133.96 (CH), and 144.48 (C),
144.27 (C3 of triazol), 153.73 (C5 of triazol).
5-Benzyl-4-[(2-hydroxyphenyl)methyliden]amino-
2-phenacyl-2,4-dihydro-3H-1,2,4-triazol-3-one (IIj);
yield 63.17%; mp 151–152°ë (from ethanol); IR: 3294
(OH), 1702 (2 C=O), 1604 and 1594 (2 C=N) cm–1; 1H
NMR: 3.86 (2 H, s, CH2), 5.42 (2 H, s, NCH2), 6.90–
6.97 (2 H, m, Ar), 7.28–7.39 (3 H, m, Ar), 7.48–7.54
(5 H, m, Ar), 8.02–8.11 (4 H, m, Ar), 9.92 (H, s,
N=CH), 10.33 (H, s, OH); 13C NMR: 36.22 (CH2),
54.04 (NCH2), Ar: 119.45 (CH), 119.92 (C), 122.01
(CH), 124.83 (2 CH), 126.24 (2 CH), 128.07 (CH),
128.66 (2 CH), 129.12 (2 CH), 131.77 (2 CH), 133.72
(CH), 134.02 (2 C), 136.18 (C), 142.52 (C3 of triazol),
152.34 (C5 of triazol), 154.23 (N=CH), 192.12
(CH2C=O).
4-Amino-5-benzyl-2-(2-hydroxy-2-phenethyl)-2,4-
dihydro-3H-1,2,4-triazol-3-one (IIIb); yield 50.23%;
mp 164–165°C; IR: 3362–3332 (OH + NH2), 1693
1
(C=O), 1630 (2 C=N) cm–1; H NMR: 3.92 (2 H, s,
CH2), 4.31 (H, d, NCH2), 4.38 (H, d, NCH2), 5.02 (H,
m, CH), 5.38 (2 H, s, NH2), 6.68 (H, br. s, OH), 7.31–
7.40 (3 H, m, Ar), 7.51–7.62 (2 H, m, Ar), 7.73–8.07
13
(5 H, m, Ar); C NMR: 34.65 (CH2), 55.45 (NCH2),
69.21 (CH), Ar: 124.18 (2 CH), 125.22 (2 CH), 126.51
(2 CH), 127.29 (2 CH), 127.93 (CH), 128.65 (CH),
130.08 (2 C), 142.15 (C3 of triazol), 152.18 (C5 of tri-
azol); MS, m/z (I, %): 311.12 [M]+ (13), 293.08 (26),
282.18 (13), 155.71 (14), 152.83 (100).
General procedure for the synthesis of 5-
alkyl[aryl]-4-Y-2-(2-hydroxy-2-phenethyl)-2,4-dihyd-
ro-3H-1,2,4-triazol-3-ones (IIIa–IIIj). A solution of
the corresponding compound (IIa)–(IIj) (0.01 mol) in
absolute ethanol (40 ml) was treated with a solution of
NaBH4 (0.03 mol) in absolute ethanol (30 ml). The
mixture was refluxed for 8 h and then poured in 500 ml
of water. A cooling in a deep freezer led to precipitation
of a solid of (IIIa)–(IIIj), which was recrystallized
from 1 : 1 ethanol–water mixture.
4-(Benzylamino)-5-methyl-2-(2-hydroxy-2-phen-
ethyl)-2,4-dihydro-3H-1,2,4-triazol-3-one
(IIIc);
yield 48.77%; mp 115–116°C; IR: 3231 (OH + NH),
1678 (C=O), 1584 and 1510 (2 C=N) cm–1; H NMR:
1
1.93 (3 H, s, CH3), 3.58 (H, d, NCH2), 3.83 (H, d,
NCH2), 4.03 (2 H, br. s, NHCH2), 5.29 (H, br. s, CH),
5.56 (H, d, J 4.0 Hz, OH), 6.52 (H, br. s, NH), 7.20–
7.52 (10 H, m, Ar); 13C NMR: 24.15 (CH3), 48.85
(NHCH2), 58.78 (NCH2), 70.46 (CH), Ar: 126.07 (2
4-Amino-2-(2-hydroxy-2-phenethyl)-5-methyl-2,4-
dihydro-3H-1,2,4-triazol-3-one (IIIa); yield 48.77%; CH), 127.25 (CH), 127.42 (CH), 129.59(CH), 128.23
mp 98–99°C (from ethanol); IR: 3365–3238 (OH + (CH), 128.64 (CH), 129.81 (CH), 129.96 (2 CH),
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DEMIRBA‹ et al.
137.14 (C), 142.65 (C), 144.13 (C3 of triazol), 152.31 2.34 (3 H, s, CH3), 3.74 (H, br. s, NHCH2), 4.24 (H, d,
(C5 of triazol); MS, m/z (I, %): 324.37 [M]+ (4), 311.12 NCH2), 4.47 (H, d, NCH2), 4.97 (H, br. s, CH), 5.47 (H,
(6), 110.98 (100).
d, J 4.2 Hz, OH), 6.19 (H, br. s, NH), 6.86–6.94 (3 H,
m, Ar), 7.02–7.13 (H, m, Ar), 7.21–7.26 (5 H, m, Ar),
9.49 (H, s, PhOH); 13C NMR: 19.65 (CH3), 47.34
(NHCH2), 56.31 (NCH2), 70.27 (CH),Ar: 116.38 (CH),
119.31 (CH), 123.62 (C), 125.24 (2CH), 128.03 (2CH),
129.28 (CH), 129.03 (CH), 130.21 (CH), 137.09 (C),
and 139.11 (C), 145.56 (C3 of triazol), 153.68 (C5 of
triazol).
4-(Benzylamino)-2-(2-hydroxy-2-phenethyl)-5-
phenyl-2,4-dihydro-3H-1,2,4-triazol-3-one (IIId);
yield 42.19%; mp 110–111°C; IR: 3251 (OH + NH),
1689 (C=O), 1583 and 1550 (2 C=N) cm–1; H NMR:
1
3.70–3.93 (2 H, dd, NCH2), 4.15 (2 H, br. s, NHCH2),
4.95 (H, m, CH), 5.62 (H, d, J 4.1 Hz, OH), 6.60 (H, br.
s, NH), 7.05–7.22 (6 H, m, Ar), 7.23–7.45 (5 H, m, Ar),
7.65–7.78 (4 H, m, Ar); 13C NMR: 49.18 (NHCH2),
52.31 (NCH2), 70.13 (CH), Ar: 126.04 (3 CH), 126.34
(C), 127.19 (3 CH), 127.27 (3 CH), 128.02 (3 CH),
128.81 (2 CH), 129.58 (CH), 136.25 (C), and 142.50
(C), 144.13 (C3 of triazol), 152.31 (C5 of triazol); MS,
m/z (I, %): 387.12 [M]+ (13), 369.11 (36), 265.07 (16),
249.10 (100), 177.03 (10).
4-(2-Hydroxybenzyl)amino-2-(2-hydroxy-2-phe-
nethyl)-5-ethyl-2,4-dihydro-3H-1,2,4-triazol-3-one
(IIIh); yield 57.30%; mp 117–119°C; IR: 3246 (2 OH +
NH), 1688 (C=O), 1593 (2 C=N); 1H NMR: 0.98 (3 H,
t, J 7.4 Hz, CH3), 2.25 (2 H, q, J 7.4 Hz, CH2), 3.60 (H,
d, NCH2), 3.85 (H, d, NCH2) 4.06 (2 H, br.s, NHCH2),
4.94 (H, br. s, CH), 5.59 (H, d, J 4.1 Hz, OH), 6.21 (H,
br. s, NH), 6.72–6.80 (3 H, m, Ar), 6.84–6.94 (H, m,
Ar), 6.98–7.11 (H, m, Ar), 7.30–7.37 (4 H, m, Ar), 9.53
(H, s, PhOH); 13C NMR: 18.63 (CH3), 25.69 (CH2),
47.33 (NHCH2), 55.81 (NCH2), 70.15 (CH), Ar: 115.14
(CH), 118.64 (2 CH), 123.09 (C), 126.17 (CH), 127.33
(2 CH), 127.59 (CH), 128.55 (CH), 133.52 (CH),
140.67 (C), 144.46 (C), 146.94 (C3 of triazol), 153.02
(C5 of triazol).
5-Benzyl-4-(2,4-dichlorobenzyl)amino-2-(2-hyd-
roxy-2-phenethyl)-2,4-dihydro-3H-1,2,4-triazol-3-one
(IIIe); yield 51.5%; mp 114–115°C; IR: 3446 (OH),
3232 (NH), 1688 (C=O), 1586 (2 C=N) cm–1; 1H NMR:
3.68 (2 H, s, CH2), 3.71 (2 H, br. s, NHCH2), 4.05 and
4.29 (2 H, dd, NCH2), 4.96 (H, t, J 12.0, CH), 5.54 (H,
br. s, OH), 6.47 (H, br. s, NH), 7.03–7.16 (3 H, m, Ar),
7.21–7.33 (5 H, m, Ar), 7.33–7.39 (5 H, m, Ar); 13C
NMR: 35.82 (CH2), 48.91 (NHCH2), 51.83 (NCH2),
4-(2-Hydroxybenzyl)amino-2-(2-hydroxy-2-phe-
70.43 (CH), Ar: 126.09 (CH), 128.63 (2 CH), 127.18 nethyl)-5-propyl-2,4-dihydro-3H-1,2,4-triazol-3-one
(3 CH), 127.98 (3 CH), 128.18 (2 CH), 128.63 (2 CH), (IIIi); yield 56.07%; mp 144–145°C; IR: 3246 (2 OH +
NH), 1684 (C=O), 1595 (2 C=N) cm–1; 1H NMR: 0.75
(3 H, t, J 7.4 Hz, CH3), 1.32–1.43 (2 H, m, CH2), 2.06
(2 H, t, J 7.6 Hz, CH2), 3.76 (2 H, br. s, NHCH2), 3.60
and 3.85 (2 H, dd, NCH2), 4.88 (H, br. s, CH), 5.52 (H,
d, J 4.0, OH), 6.19 (H, br. s, NH), 6.69–6.78 (3 H, m,
Ar), 7.05–7.09 (H, m, Ar), 7.18–7.28 (5 H, m, Ar), 9.47
(H, s, PhOH); 13C NMR: 13.38 (CH3), 18.63 (CH2),
25.66 (CH2), 47.37 (NHCH2), 55.92 (NCH2), 70.15
(CH), Ar: 114.88 (CH), 118.58 (CH), 123.08 (C),
126.03 (2 CH), 127.17 (2 CH), 127.97 (CH), 128.53
(CH), 130.70 (CH), 142.66 (C), 145.68 (C), 146.96 (C3
of triazol), 152.44 (C5 of triazol); MS, m/z (I, %):
369.02 [M]+ (27), 351.08 (17), 285.90 (15), 284.96
(100), 248.96 (44), 244.98 (32), 214.09 (20), 121.80
(13), 105.88 (11).
132.70 (C), 132.99 (C), 133.50 (C), 134.34 (C), 135.49
(C), 145.78 (C3 of triazol), 152.68 (C5 of triazol); MS,
m/z (I, %): 504.98 [M + 1]+ (25), 486.97 (44), 475.24
(57), 454.22 (14), 453.22 (52), 249.02 (39), 227.08
(43), 209.02 (12), 164.90 (13), 116.96 (13), 113.95
(20), 100.11 (29), 79.18 (100), 60.32 (47).
5-(4-Chlorobenzyl)-4-(2,4-dichlorobenzyl)ami-
no-2-(2-hydroxy-2-phenethyl)-2,4-dihydro-3H-1,2,4-
triazol-3-one (IIIf); yield 50.32%; mp 126–127°C; IR:
3263–3084 (OH + NH), 1692 (C=O), 1612 and 1602
(2 C=N) cm–1; 1H NMR: 3.48 (2 H, s, CH2), 3.63–3.89
(2 H, m, NCH2), 4.07 (H, s, NHCH2), 4.89 (H, br. s,
CH), 5.63 (H, d, J 4.0 Hz, OH), 6.64 (H, br. s, NH),
7.00–7.05 (3 H, m, Ar), 7.16–7.33 (7 H, m, Ar), 7.53 (2
H, br. s, Ar); 13C NMR: 34.12 (CH2), 48.88 (NHCH2),
54.85 (NCH2), 70.11 (CH), Ar: 126.25 (2 CH), 127.18
(CH), 127.24 (2 CH), 127.99 (CH), 128.07 (2 CH),
128.64 (CH), 129.13 (2 CH), 130.11 (CH), 131.17 (C),
132.69 (C), 133.04 (C), 133.44 (2CH), 134.34 (C)],
145.37 (C3 of triazol), 152.21 (C5 of triazol); MS, m/z
(I, %): 504.98 [M + 1]+ (25), 486.97 (44), 475.24 (57),
454.22 (14), 453.22 (52), 249.02 (39), 227.08 (43),
209.02 (12), 164.90 (13), 116.96 (13), 113.95 (20),
100.11 (29), 79.18 (100), 60.32 (47).
5-Benzyl-4-(2-hydroxybenzyl)amino-2-(2-hydro-
xy-2-phenethyl)-2,4-dihydro-3H-1,2,4-triazol-3-one
(IIIj); yield 47.60%; mp 139–140°C; IR: 3296 (2 OH),
3239 (NH), 1686 (C=O), 1609 (2 C=N) cm–1; 1H NMR
3.78 (2 H, s, CH2), 3.76 (H, br.s, NHCH2), 4.06 and
4.31 (2 H, dd, NCH2), 5.11 (H, t, J 12.0 Hz, CH), 5.49
(H, br. s, OH), 6.29 (H, br. s, NH), 6.71–6.82 (3 H, m,
Ar), 7.09–7.12 (2 H, m, Ar), 7.16–7.25 (4 H, m, Ar),
7.28–7.36 (5 H, m, Ar), 9.54 (H, s, PhOH); 13C NMR:
4-(2-Hydroxybenzyl)amino-2-(2-hydroxy-2-phe- 34.11 (CH2), 48.12 (NHCH2), 49.09 (NCH2), 71.06
nethyl)-5-methyl-2,4-dihydro-3H-1,2,4-triazol-3-one
(IIIg); yield 45.16%; mp 154–155°C; IR: 3341 (2 OH),
(CH), Ar: 121.81 (CH), 122.26 (2 CH), 123.66 (2 CH),
124.07 (C), 126.05 (CH), 126.17 (2 CH), 127.94
3256 (NH), 1684 (C=O), 1595 (2 C=N) cm–1; 1H NMR: (2 CH), 128.53 (2 CH), 130.72(2 CH), 136.14 (C),
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141.97 (C), 143.12 (C), 145.45 (C3 of triazol), 155.71 6.11 (H, br. s, NH), 6.84–6.92 (3 H, m, Ar), 6.98–7.09
(C5 of triazol); MS, m/z (I, %): 417.06 [M]+ (20),
(2 H, m, Ar), 7.11–7.25 (5 H, m, Ar), 7.28–7.46 (3 H,
m, Ar), 7.51–7.56 (3 H, m, Ar), 8.01–8.07 (2 H, m, Ar);
381.17 (18), 380.16 (49), 364.16 (11), 305.17 (22),
304.17 (100), 302.16 (18), 282.14 (72), 265.07 (13), 13C NMR: 21.98 (CH3), 35.21 (PhCH2), 44.35
256.79 (35), 240.85 (22), 228.05 (16), 198.81 (13),
176.78 (19), 160.78 (13), 158.77 (33), 152.94 (79),
142.71 (30), 141.77 (16), 119.80 (29), 110.77 (33),
100.79 (64).
A general procedure for the synthesis of com-
pounds (IV)–(VII). A solution of the corresponding
compound (IIg), (IIj), or (IIIj) (0.01 mol) in ethanol
[dry toluene for (IIIb)] was refluxed with equivalent
amount of NaOH [sodium metal for (IIIb)] for 1 h.
Then, p-toluenesulfonyl chloride (0.01 mol) was added,
and the reaction mixture was refluxed for an additional
5 h. After the removal of solvents at a reduced pressure,
a solid was obtained, which was recrystallized from
ethanol.
(NHCH2), 51.51 (NCH2), 73.28 (CH), Ar: 121.70 (C),
125.93 (CH), 126.12 (CH), 126.84 (2 CH), 127.05
(2 CH), 127.76 (2 CH), 127.85 (2 CH), 127.92 (2 CH),
128.16 (2 CH), 129.17 (4 CH), 129.66 (2 C), 134.62
(C), 134.79 (C), 141.11 (C), 143.17 (C3 of triazol),
155.67 (C5 of triazol); MS, m/z (I, %): 569.82 [M]+
(100), 567.87 (20), 537.85 (43), 304.18 (24), 282.20
(30), 276.98 (18), 274.97 (19), 218.88 (13), 200.85
(16), 155.93 (47), 116.87 (16).
4-Amino-5-benzyl-2-(2-tosyloxy-2-phenethyl)-2,4-
dihydro-3H-1,2,4-triazol-3-one (VII); yield: 54.36%;
mp 167–169°C; IR: 3352–3318 (NH2), 1698 (C=O),
1612 (2 C=N) cm–1; 1H NMR: 2.38 (H, s, CH3), 3.92 (2
H, s, PhCH2), 4.01 and 4.25 (2 H, dd, NCH2), 5.39 (2
H, s, NH2), 5.53 (H, m, CH), 6.69 (H, br. s, OH), 7.24–
7.38 (3 H, m, Ar), 7.48–7.60 (2 H, m, Ar), 7.71–7.97
5-Methyl-4-[(2-tosyloxyphenyl)methyliden]ami-
no-2-phenacyl-2,4-dihydro-3H-1,2,4-triazol-3-one
(IV); yield: 48.28%, mp 181–182°C; IR: 1719 and
13
1
(5 H, m, Ar); C NMR: 22.32 (CH3), 35.23 (PhCH2),
1693 (C=O), 1615 and 1594 (C=N) cm–1; H NMR:
49.56 (NCH2), 72.21 (CH), Ar: 126.31 (2 CH), 127.32
(2 CH), 128.35 (2 CH), 129.29 (2 CH), 130.84 (CH),
133.58 (CH), 141.00 (2 C), 143.82 (C3 of triazol),
145.61 (C5 of triazol).
1.88 (3 H, s, CH3), 2.47 (3 H, s, CH3), 5.28 (2 H, s,
NCH2), 7.08–7.14 (3 H, m, Ar), 7.17–7.25 (2 H, m, Ar),
7.33–7.39 (6 H, m, Ar), 7.45–7.62 (2 H, m, Ar), 9.03
(H, s, N=CH); 13C NMR: 12.34 (CH3), 22.15 (CH3),
48.31 (NCH2), Ar: 119.36 (CH), 119.48 (CH), 121.84
(2 CH), 123.36 (CH), 124.73 (C), 125.95 (2 CH),
127.72 (CH), 128.37 (2 CH), 130.02 (3 CH), 131.71 (2
C), 139.12 (C), 145.93 (C), 143.17 (C3 of triazol),
145.65 (C5 of triazol), 155.12 (N=CH), 192.48 (C=O);
MS, m/z (I, %): 491.21 [M]+ (100), 320.23 (10), 305.19
(11), 304.18 (52), 283.23 (9), 282.22 (37), 156.86 (11),
155.86 (17), 152.96 (22).
A general procedure for the synthesis of com-
pounds (VIII)–(XI). A solution of the corresponding
compound (II) or (III) (0.01 mol) in ethanol [(IIg) and
(IIIg)] or dry toluene [(IIIa) and (IIId)] was refluxed
with equivalent amount of NaOH [(IIg) and (IIIg)] or
sodium metal [(III) and (IIId)] for 1 h. Then, benzyl
bromide (0.01 mol) was added and reflux was pro-
longed for additional 5 h. After the removal of solvents
at a reduced pressure, a solid was obtained, which was
recrystallized from ethanol.
5-Benzyl-4-[(2-tosyloxyphenyl)methyliden]ami-
no-2-phenacyl-2,4-dihydro-3H-1,2,4-triazol-3-one (V);
yield: 47.85%; mp 141–142°C; IR: 1715 and 1684 (2
C=O), 1623 and 1597 (C=N) cm–1; 1H NMR: 2.33 (3 H,
s, CH3), 3.94 (2 H, s, CH2), 5.39 (2 H, s, NCH2), 6.79–
6.93 (3 H, m, Ar), 6.97–7.08 (2 H, m, Ar), 7.18–7.25 (9
H, m, Ar), 7.32–7.47 (4 H, m, Ar), 9.21 (H, s, N=CH);
13C NMR: 21.66 (CH3), 33.59 (CH2), 51.67 (NCH2),
Ar: 120.52 (C), 122.18 (2 CH), 125.34 (2 CH), 126.82
(2 CH), 127.31 (2 CH), 128.29 (2 CH), 128.63 (2 CH),
129.42 (2 CH), 129.76 (2 CH), 133.71 (C), 137.12 (2
CH), 138.43 (C), 139.12 (CH), 141.61 (C), 142.56 (C),
143.14 (C3 of triazol), 145.52 (C5 of triazol), 152.65
(N=CH), 194.45 (C=O); MS, m/z (I, %): 567.12 [M]+
(17), 304.19 (11), 282.10 (7), 153.91 (15), 152.90
(100).
4-[(2-Benzyloxyphenyl)methyliden]amino-5-me-
thyl-2-phenacyl-2,4-dihydro-3H-1,2,4-triazol-3-one
(VIII); yield 47.63%; mp 179–180°C; IR: 1721 and
1684 (2 C=O), 1612 and 1597 (2 C=N) cm–1; 1H NMR
1.99 (3 H, s, CH3), 4.44 (2 H, s, NCH2), 4.64 (2 H, s,
OCH2), 6.89–6.93 (2 H, m, Ar), 7.13–7.21 (2 H, m, Ar),
7.28–7.35 (3 H, m, Ar), 7.46–7.62 (4 H, m, Ar), 7.67–
7.84 (3 H, m, Ar), 9.72 (H, s, N=CH); 13C NMR: 13.26
(CH3), 49.45 (NCH2), 68.13 (OCH2), Ar: 114.61 (2
CH), 118.75 (2 CH), 120.14 (2 CH), 123.66 (CH),
125.17 (CH), 127.34 (CH), 128.21 (CH), 128.66 (C),
129.48 (2 CH), 132.11 (CH), 134.05 (C), 136.53 (CH),
139.16 (C), 142.11 (C), 144.16 (C3 of triazol), 154.53
(C5 of triazol), 163.25 (N=CH), 194.08 (C=O); MS,
m/z (I, %): 427.11 [M]+ (100), 375.00 (13), 337.04 (13),
304.16 (18), 282.17 (31), 156.84 (14), 152.96 (59),
142.81 (24).
5-Benzyl-2-(2-hydroxy-2-phenethyl)-4-[(2-tosyl-
oxybenzyl)amino]-2,4-dihydro-3H-1,2,4-triazol-3-
one (VI); yield 66.13%; mp 124–125°C; IR: 3268
(OH), 1709 and 1691 (2 C=O), 1594 and 1581 (2 C=N)
4-[2-(Benzyloxy)benzyl]amino-2-(2-hydroxy-2-
phenethyl)-5-methyl-2,4-dihydro-3H-1,2,4-triazol-
3-one (IX); yield: 42.54%; mp 132–134°C; IR: 3324
(OH), 3256 (NH), 1693 (C=O), 1598 (2 C=N) cm–1; 1H
1
cm–1; H NMR: 2.24 (3 H, s, CH3), 3.62 (2 H, s,
PhCH2), 3.55 and 3.70 (2 H, dd, NCH2), 4.26 (H, br. s,
NHCH2), 5.74 (H, br. s, CH), 5.98 (H, d, J 4.0, OH),
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 4 2005

396
DEMIRBA‹ et al.
NMR: 2.01 (3 H, s, CH3), 4.16 (H, m, NHCH2), 3.88 Candida glabrata ATCC 66032. All the newly synthe-
sized compounds were dissolved in DMSO and acetone
and 95% ethanol to prepare stock solutions with
100-mg/ml concentration.
and 4.13 (2 H, dd, NCH2), 4.99 (H, br. s, CH), 5.14
(2 H, s, OCH2), 5.57 (H, br. s, OH), 6.83 (H, br. s, NH),
6.89–6.93 (3 H, m, Ar), 7.11–7.17 (H, m, Ar), 7.19–
7.38 (10 H, m, Ar); 13C NMR: 21.01 (CH3), 44.98
The antimicrobial effects of the substances were
(NHCH2), 54.24 (NCH2), 70.99 (CH), 72.01 (OCH2), tested quantitatively in respective broth media by using
double dilution, and the minimal inhibition concentra-
tion (MIC) values (µg/ml) were determined [21]. The
antibacterial and antifungal assays were performed in
the Mueller–Hinton broth (MH) (Difco, Detroit, MI) at
pH 7.3 and buffered Yeast Nitrogen Base (Difco,
Detroit, MI) at pH 7.0, respectively. The MIC was
defined as the lowest concentration that showed no
growth. Ampicillin and fluconazole were used as stan-
dard antibacterial and antifungal drugs, respectively.
DMSO, acetone and 95% ethanol were used as solvent
controls. The results are given in Table 1.
Ar: 119.28 (2 CH), 121.96 (CH), 124.36 (C), 126.21
(4 CH), 129.45 (3 CH), 129.91 (2 CH), 130.42 (2 CH),
132.21 (C), 138.23 (C), 141.67 (C), 143.60 (C3 of tria-
zol), 155.43 (C5 of triazol); MS, m/z (I, %) 431.12 [M]+
(100), 413.10 (24), 410.10 (18), 364.05 (16), 363.05
(69), 342.07 (9), 341.07 (46), 324.05 (11), 323.04 (53),
307.03 (16), 304.14 (13), 282.09 (18), 256.97 (14),
216.91 (41), 196.87 (10), 158.75 (11), 152.91 (36),
114.86 (29).
4-Amino-2-[2-(benzyloxy)-2-phenethyl]-5-methyl-
2,4-dihydro-3H-1,2,4-triazol-3-one
(X);
yield:
58.38%; mp 139–140°C; IR: 3334–3218 (NH2), 1693
Antitumor screening studies. The screening
experiments were carried out by the Developmental
Therapeutic Program of the National Cancer Institute
(NCI), Bethesda, Maryland, United States. Fourteen
compounds (IIa), (IIb), (IIg), (IIf), (IIi), (IIIb), (IIIf),
(IIIi), (IV), (V), (VI), (VII), (VIII) and (IX) were
selected by NCI for screening toward 3 human tumor
cell lines: vic., breast cancer (MCF7), non small cell
lung cancer (NCI-H460) and CNS (SF-268). Each cell
line was inoculated and preincubated on a microtiter
plate. Test agents were then added at a single concen-
tration and culture incubated for 48 h. End point deter-
minations were made with Alamar Blue [22]. The
screening results are summarized in Table 2. Results for
each test agent are reported as the percent of growth of
the treated cells compared with the untreated control
cells. The compounds that reduce the growth any one of
(C=O), 1617 and 1543 (2 C=N) cm–1; H NMR: 1.96
1
(3 H, s, CH3), 3.52 and 3.79 (2 H, dd, NCH2), 4.67 (2
H, s, OCH2), 5.43 (H, br. s, CH), 5.56 (H, br. s, OH),
6.56 (H, s, NH), 7.21–7.34 (6 H, m, Ar), 7.42–7.49
(2 H, m, Ar), 7.51–7.56 (2 H, m, Ar); 13C NMR: 23.05
(CH3), 57.61 (NCH2), 73.34 (CH), Ar: 126.35 (2 CH),
127.11 (CH), 128.93 (CH), 129.19 (CH), 129.75 (CH),
130.63 (CH), 130.91 (CH), 135.25 (2 CH), 138.91 (C),
145.26 (C), 143.62 (C3 of triazol), 154.11 (C5 of tria-
zol).
4-Benzylamino-2-[2-(benzyloxy)-2-phenethyl]-5-
phenyl-2,4-dihydro-3H-1,2,4-triazol-3-one
(XI);
yield 54.87%; mp 188–189°C; IR: 3251 (NH), 1702
(C=O), 1597 and 1508 (C=N) cm–1; 1H NMR: 3.36 (H,
br. s, NHCH2), 3.62–3.81 (2 H, dd, NCH2), 4.27 (2 H,
s, OCH2), 4.92 (H, t, J 12.0 Hz, CH), 6.12 (H, t, J the cell lines to approximately 32% or less were
regarded as having antitumor activity.
4.0 Hz, NH), 6.73 (3 H, br. s, Ar), 6.91–7.06 (3 H, m,
Ar), 7.17–7.47 (8 H, m, Ar), 7.51–7.66 (4 H, m, Ar),
8.01–8.05 (2 H, m,Ar); 13C NMR: 64.05 (OCH2), 49.27
(NHCH2), 52.00 (NCH2), 79.27 (CH), Ar: 125.72 (C),
126.31 (2 C), 127.06 (2 CH), 127.27 (2 CH), 127.71 (2
CH), 127.81 (2 CH), 128.09 (2 CH), 128.22 (CH),
128.69 (2 CH), 128.87 (CH), 129.00 (2 CH), 133.43 (2
CH), 134.66 (C), 135.75 (C), 137.09 (C), 145.21 (C3 of
triazol), 152.13(C5 of triazol); MS, m/z (I, %): 477.07
[M]+ (6), 476.13 (29), 475.06 (100), 454.17 (8), 453.16
(27), 100.06 (19), 91.01 (19), 79.01 (50), 64.06 (79).
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