Synthesis and biological activities of novel 1,2,4-triazole thiones and bis(1,2,4-triazole thiones) containing phenylpyrazole and piperazine moieties

Article abstract of DOI:10.1016/j.jfluchem.2016.02.004

A series of novel 1,2,4-triazole thione and bis(1,2,4-triazole thione) derivatives containing phenylpyrazole and substituted piperazine moieties have been conveniently synthesized via Mannich reaction using various 1,2,4-triazole thiols, substituted piperazines, and formaldehyde at room temperature in short time. Their structures were confirmed by IR, ¹H NMR, ¹³C NMR and elemental analysis. The preliminary bioassays for 27 new title compounds have shown that some of them possess certain herbicidal activities against Echinochloa crusgalli at 100 μg/mL concentration. Some of the compounds exhibited significant in vitro fungicidal activities, especially against Cercospora arachidicola and Rhizoctonia cerealis at the concentration of 50 μg/mL, and were comparable with that of control Triadimefon. Meanwhile, IXp held the control efficacy of 60% against Puccinia sorghi Schw. at 200 μg/mL concentration in the in vivo test. The SAR analysis has indicated that compounds with two CF₃ groups both on triazole and pyrazole rings are more effective than compounds with one CF₃ group or two CH₃ groups according to their fungicidal activity data. On the whole, compounds VIIIg, IXi, IXo and IXp could be used as novel lead structures for the design and discovery of new fungicides.

Full text of DOI:10.1016/j.jfluchem.2016.02.004

Accepted Manuscript
Title: Synthesis and biological activities of novel 1,2,4-triazole
thiones and bis(1,2,4-triazole thiones) containing
phenylpyrazole and piperazine moieties
Author: Bao-Lei Wang Li-Yuan Zhang Yi-Zhou Zhan Yan
Zhang Xiao Zhang Li-Zhong Wang Zheng-Ming Li
PII:
S0022-1139(16)30032-X
DOI:
Reference:
http://dx.doi.org/doi:10.1016/j.jfluchem.2016.02.004
FLUOR 8731
To appear in:
FLUOR
Received date:
Revised date:
Accepted date:
3-12-2015
12-2-2016
14-2-2016
Please cite this article as: Bao-Lei Wang, Li-Yuan Zhang, Yi-Zhou Zhan,
Yan Zhang, Xiao Zhang, Li-Zhong Wang, Zheng-Ming Li, Synthesis and
biological activities of novel 1,2,4-triazole thiones and bis(1,2,4-triazole thiones)
containing phenylpyrazole and piperazine moieties, Journal of Fluorine Chemistry
http://dx.doi.org/10.1016/j.jfluchem.2016.02.004
This is a PDF file of an unedited manuscript that has been accepted for publication.
As a service to our customers we are providing this early version of the manuscript.
The manuscript will undergo copyediting, typesetting, and review of the resulting proof
before it is published in its final form. Please note that during the production process
errors may be discovered which could affect the content, and all legal disclaimers that
apply to the journal pertain.

Synthesis and Biological Activities of Novel 1,2,4-Triazole
Thiones and Bis(1,2,4-Triazole Thiones) Containing
Phenylpyrazole and Piperazine Moieties
Bao-Lei Wang*, Li-Yuan Zhang, Yi-Zhou Zhan, Yan Zhang, Xiao Zhang, Li-Zhong
Wang, Zheng-Ming Li*
State Key Laboratory of Elemento-Organic Chemistry, Collaborative Innovation
Center of Chemical Science and Engineering, Nankai University, Tianjin 300071, PR
China.
*Corresponding author. E-mail addresses: nkwbl@nankai.edu.cn (B.-L. Wang),
nkzml@vip.163.com (Z.-M. Li).
1

Graphical abstract
A series of novel phenylpyrazole- and piperazine-containing (bis)1,2,4-triazole
thiones were synthesized via Mannich reaction in high yields. The compounds with
two CF3 groups exhibited excellent fungicidal activities.
Highlights
 27 novel phenylpyrazole- and piperazine-containing (bis)1,2,4-triazole thiones
were synthesized.
 Several compounds exhibited significant fungicidal activities.
 The compounds with two CF3 groups on triazole and pyrazole rings had excellent
fungicidal activities.
 VIIIg, IXi, IXo and IXp could be used as new fungicide lead compounds.
2

Abstract: A series of novel 1,2,4-triazole thione and bis(1,2,4-triazole thione)
derivatives containing phenylpyrazole and substituted piperazine moieties have been
conveniently synthesized via Mannich reaction using various 1,2,4-triazole thiols,
substituted piperazines, and formaldehyde at room temperature in short time. Their
1
structures were confirmed by IR, H NMR, 13C NMR and elemental analysis. The
preliminary bioassays for 27 new title compounds have shown that some of them
possess certain herbicidal activities against Echinochloa crusgalli at 100 μg/mL
concentration. Some of the compounds exhibited significant in vitro fungicidal
activities, especially against Cercospora arachidicola and Rhizoctonia cerealis at the
concentration of 50 μg/mL, and were comparable with that of control Triadimefon.
Meanwhile, IXp held the control efficacy of 60% against Puccinia sorghi Schw. at
200 μg/mL concentration in the in vivo test. The SAR analysis has indicated that
compounds with two CF3 groups both on triazole and pyrazole rings are more
effective than compounds with one CF3 group or two CH3 groups according to their
fungicidal activity data. On the whole, compounds VIIIg, IXi, IXo and IXp could be
used as novel lead structures for the design and discovery of new fungicides.
Keywords: Trifluoromethyl; pyrazole; triazole thione; biological activities
3

1. Introduction
Fluorine- and heterocycle-containing compounds play a very important role not
only in the research area of organic chemistry, but also in a variety of applied
chemistry areas, such as pharmaceutical chemistry, pesticidal chemistry and material
science [1-9]. It is known that the introduction of fluorine atoms or trifluoromethyl
groups may increase the biological activity of the parent compounds. Many
trifluoromethyl-containing compounds were found to have biological activities as
fungicides [10], insecticides [11], herbicides [12], COX-2 inhibitors [13], CETP
inhibitors [14], anti-inflammatory agents [15, 16] and anti-cancer agents [17, 18]. For
example, Flubendiamide, a new insecticide which targets the insect ryanodine recepor,
found by Nippon Kayaku Co., has a heptafluoroisopropyl group (two trifluoromethyl
and one fluorine atom) on one of the benzene rings of its phthalic diamide structure
[19]. Furthermore, azole thiones as one kind of heterocyclic compounds have
attracted much attention due to their interesting structural properties and versatile
pharmacological activities; many of them have been found to possess antituberculous
[20], antimalarial [21], antifungal [22] and anticancer [23] activities.
Moreover, the pyrazole and its derivatives are known to occupy an important
position in the field of medicinal and pesticidal chemistry; they are always associated
with various pharmacological and biological activities. As medicines, many of them
possess significant antibacterial, antiviral, anticoagulant, antimicrobial,
anti-inflammatory and anticancer properties [24-26]. Their pesticidal activities are
manifested by their use as insecticides, fungicides, herbicides and plant growth
regulators [27, 28]. For example, Fipronil, an insecticide that is used to control
lepidopteran and coleopteran insects, is a kind of pyrazole derivative [29].
In our previous work, some triazole thiones with trifluoromethyl and piperazine
groups were synthesized successfully and found to have favorable herbicidal and
significant fungicidal activities [30, 31], which provided an important reference for us
to make some further structural modifications. The preliminary SAR studies have
illustrated the necessity for further optimization based on the general structure of this
type of compounds, especially for the phenyl group and the substituent on the
4

piperazine ring part [30, 31]. Considering all that mentioned above and with our
pursuit to look for novel heterocyclic compounds with pesticidal potential, in this
paper we report the synthesis of a series of novel 1,2,4-triazole thione derivatives with
phenylpyrazole and substituted benzylpiperazine or various arylpiperazine moieties,
as well as bis(1,2,4-triazole thione) derivatives with phenylpyrazole and piperazine
moieties via Mannich reaction. For SAR study, the methyl and trifluoromethyl groups
were introduced into the structures in various combinations on both triazole and
pyrazole rings. These novel compounds were further evaluated for heribicidal and
fungicidal activities.
To the best of our knowledge, the synthesis and pesticidal activities of the title
compounds with such structural characteristics have not been studied so far. Thus, the
research in this paper may provide usful information for the synthesis and discovery
of new bioactive fluorine and heterocycle-containing compounds.
2. Results and discussion
2.1 Synthesis
The synthetic routes of the intermediates and the title compounds are shown in
Scheme 1, Scheme 2 and Scheme 3.
The trifluoromethyl-containing carbonyl compounds, such as trifluoroacetophenone,
ethyl trifluoroacetoacetate and 1,1,1,5,5,5-hexafluoro-2,4-pentanedione are known as
important building blocks for the construction of trifluoromethyl imines or enamines
in either open-chain or heterocyclic form, via the condensation reaction with amines
[32-36]; however, the condensation of ethyl trifluoroacetoacetate (or ethyl
acetoacetate) with substituted hydrazines will result in 5-trifluoromethyl(or
5-methyl)-pyrazolone compounds [37, 38]. The latter method was used to smoothly
synthesize intermediate IV. Phenylpyrazole carbaldehyde V was prepared via
Vilsmeier reaction [39] from phenylpyrazolone IV in POCl3/DMF system in 86% ~
92% yield, followed by the condensation with amino-triazole thiol VI under refluxing
condition in absolute ethanol using acetic acid as a catalyst to give
phenylpyrazole-containing triazole thiol (Schiff base) VII in 71% ~ 87% yield.
5

Taking the thioamide structure (C(=S)-NH-) into account, it is obvious that the
intermediate VII can exist either in thiol or thione tautomeric forms. Based on the
results of our experiment, it was found that the thione isomer undergoes subsequent
Mannich reaction via the N-H at α- position of thiocarbonyl (C=S). As a result, the
condensation fo intermediate VII with formaldehyde and 4-(substituted
benzyl)piperazine I, or 4-(substituted pyrimidyl)/ phenyl/pyridylpiperazine III in
ethanol at room temperature led to novel 1,2,4-triazole thiones VIII and IX in
satisfactory yields (59% ~ 85%), respectively. Under the similar reaction conditions
using excess formaldehyde and 2 : 1 molar ratio of Schiff base VII and piperazine,
bis(1,2,4-triazole thione) X was successfully synthesized in high yield (over 82%).
Overall, this approach towards the synthesis of the title compounds have noticeable
advantages, such as, mild reaction conditions, high yield and short reaction time (1 ~
3 h).
1
13
The title compounds were identified by IR, H NMR, and C NMR spectra. The
measured elemental analyses were also consistent with the corresponding calculated
values. In IR spectra of the compounds, the stretching vibration absorption peaks of
1
C=S and C=N appeared at 1143-1177 cm−1 and 1577-1615 cm−1 respectively. In H
NMR, the -N=CH- proton showed up at δ 9.96-11.02. The piperazine ring proton
signals of compounds VIII and IX were observed at two positions; however, due to
the symmetrical structure of compound X, they were observed as a singlet at δ
2.85-2.89. In the 13C NMR spectra of representive compounds, the typical carbon
signal at δ 162.6-165.0 was derived from the resonance of thiocarbonyl group (C=S).
The piperazine carbons of compound X also appeared as one signal at δ ~ 50.3 as
opposed to two signals in compounds VIII and IX. In addition, due to the “F”
splitting, the signals of CF3 carbon and carbons adjacent to CF3 group (heterocyclic
carbon) were split into quartet or multiplet.
2.2 The biological activities
As shown in Table 1 (some data are listed in Table S1 in Supplementary
Information), compared with commercial herbicide Chlorsulfuron, most of the title
6

compounds at the concentration of 100 μg/mL showed rather weak herbicidal
activities based on the rape (Brassica campestris) root tests. However, in the case of
barnyardgrass (Echinochloa crusgalli) cup tests, at the concentration of 100 μg/mL,
several compounds showed higher herbicidal activities than the control Chlorsulfuron
(29.9%), such as VIIIc (30.0%), IXe (40.0%) and IXf (50.0%). On the whole, the title
compounds may be more effective against monocotyledonous weeds (Echinochloa
crusgalli).
The in vitro fungicidal activity data of 1,2,4-triazole thiones VIII, IX and
bis(1,2,4-triazole thiones) X for inhibition of mycelial growth in six test fungi are
listed in Table 2 (some data could be found in Table S2 in Supplementary
Information). It was found that at the test concentration of 50 μg/mL, most of the
compounds exhibited significant fungicidal activities against Alternaria solani
Sorauer, Physalospora piricola and Rhizoctonia cerealis while some of these results
were comparable with those of the contol Triadimefon. In addition, several
compounds displayed favorable actities against other test fungi, such as compound
IXg (62.5%) against Gibberella sanbinetti, compounds VIIIg (75.0%), IXo (87.5%)
and IXp (75.0%) against Cercospora arachidicola, were more effective than
Triadimefon, respectively.
According to the in vitro fungicidal activity data, several trifluoromethyl-containing
compounds were chosen to be tested for in vivo fungicidal activity against Puccinia
sorghi Schw. at 200 μg/mL concentration. From Table 3, it was shown that
compounds VIIIg, IXm, IXn and IXp have certain in vivo fungical activities,
especially IXp exhibited good activity against Puccinia sorghi Schw. with control
efficacy of 60%.
In general, these 1,2,4-triazole Mannich bases containing phenylpyrazole and
substituted piperazine moieties held significant fungicidal activities against various
7

plant fungi, while the CF3 group made an apparent influence on their fungicidal
activities, which could be well reflected particularly in the results of Rhizoctonia
cerealis test. By analyzing the SAR based on the fungicidal data against Rhizoctonia
cerealis, it was found that in most cases trifluoromethyl-containing compounds
exhibit higher activity than methyl-containing ones, and compounds with two CF3
groups both on triazole and pyrazole rings are more effective than compounds with
one CF3 (on the triazole ring, or on the pyrazole ring) or with two CH3 groups. In the
case of compounds VIII (Table 2), when R1 is fixed as Cl atom, the activity sequence
is VIIIg (A=CF3, B=CF3) > VIIId (A=CF3, B=CH3) > VIIIf (A=CH3, B=CF3) >
VIIIb (A=CH3, B=CH3); in the case of compounds IX (Table 2), when the
substituent at 4- position of piperazine is fixed as pyridyl, the activity sequence is IXn
(A=CF3, B=CF3) >> IXf (A=CF3, B=CH3) > IXj (A=CH3, B=CF3) > IXb (A=CH3,
B=CH3). For the phenyl substituent, the sequence is similar to that of pyridyl. When
the substituent at 4- position of piperazine is fixed as 4-methylpyrimidinyl, the
activity sequence is IXo (A=CF3, B=CF3) > IXg (A=CF3, B=CH3) > IXc (A=CH3,
B=CH3) > IXk (A=CH3, B=CF3). Compounds with 4,6-dimethylpyrimidinyl
substitutient show the similar trend; in the case of compounds X (Table 2), when
there are four CF3 groups in total attached to both triazole and pyrazole rings as in the
structure Xd, the activity sequence is Xd (A=CF3, B=CF3) >> Xa (A=CH3, B=CH3) ≈
IXb (A=CF3, B=CH3) ≈ Xc (A=CH3, B=CF3). In addition, according to the in vivo
fungicidal test results against Puccinia sorghi Schw., the superiority of
two-CF3-containing compounds can also be well reflected (such as IXp, IXn vs.
VIIId).
It was worthy of note that against some test fungi, VIIIg, IXi, IXo and IXp overall
exhibited higher and wider fungicidal activities than others, and were comparable
with that of control fungicide Triadimefon. Especially, VIIIg, IXo and IXp had
excellent fungicial activities against Rhizoctonia cerealis that is a very popular fungus
and creates big threat to the wheat yield in china and some other countries [40].
Therefore, they could be used as novel lead structures for the design and discovery of
new fungicides.
8

3. Conclusion
In conclusion, series of novel 1,2,4-triazole thione derivatives and bis(1,2,4-triazole
thione) derivatives with phenylpyrazole and substituted piperazine moieties have been
synthesized via Mannich reaction from the intermediate pyrazole-containing
1,2,4-triazole Schiff bases at room temperature in short time. Among 27 title
compounds, the preliminary bioassays for some of them showed promising herbicidal
activities against Echinochloa crusgalli at 100 μg/mL concentration. Some of
compounds exhibited significant in vitro fungicidal activities at 50 μg/mL
concentration against six plant fungi, especially Cercospora arachidicola and
Rhizoctonia cerealis. On the whole, coumpounds VIIIg, IXi, IXo and IXp showed
higher and wider fungicidal activities and were comparable with that of control
Triadimefon. Meanwhile, the in vivo fungicidal activity of IXp against Puccinia
sorghi Schw. at 200 μg/mL concentration was also favourable. The SAR analysis
indicated that compounds with two CF3 groups on both triazole and pyrazole rings are
more effective than those ones with one CF3 or with two CH3 groups according to
their fungicidal activity data. Overall VIIIg, IXi, IXo and IXp could be used as novel
lead structures for the design and discovery of new fungicides.
4. Experimental
4.1 Instruments and materials
The melting points were determined on a Beijing Tech X-4 binocular microscope
apparatus and are uncorrected. Infrared spectra were recorded on a Nicolet
MAGNA-560 spectrophotometer as KBr tablets. 1H NMR spectra were measured on a
Bruker AC-P500 instrument (400MHz) using TMS as an internal standard and
DMSO-d6 or CDCl3 as solvent. Elemental analysis was performed on a Vario EL
elemental analyzer. Partial 4-substituted piperazine intermediates were purchased
from Aladin and Alfa Aesar reagent companies. Anhydrous piperazine and other
materials were purchased from Nanjing Duodian Reagent Co. Ltd. All solvents were
dried in advance by using standard methods and distilled before use.
9

4.2 Synthetic procedures
4-Amino-5-methyl/trifluoromethyl-4H-1,2,4-triazole-3-thiol VIa and VIb were
prepared according to the reported procedure [41].
4.2.1 Preparation of 4-(substituted benzyl)piperazine (I)
Following the method in Lit. [42], to a solution of anhydrous piperazine (50 mmol)
in 95% ethanol (20 mL) was added concentrated hydrochloric acid (25 mmol). The
mixture was stirred under reflux and substituted benzyl chloride (25 mmol) was added
dropwise over 5 min. The mixture was refluxed for 4 ~ 8 h with TLC monitoring, then
left overnight at room temperature. The solid precipitate was filtered off and washed
with ethanol; the filtrate was evaporated in vacuo. The residue was dissolved in
saturated K2CO3 aq. (30 mL) and extracted with chloroform (5 × 8 mL). The
combined organic layer was dried over anhydrous Na2SO4 and concentrated. The
residue was then distilled under reduced pressure to give compound I as a colorless
liquid.
Ia: yield 58%, bp 131-134 oC/10mmHg; Ib: yield 37%, bp 147-150 oC/6 mmHg.
4.2.2 Preparation of 4-(4,6-disubstituted pyrimidin-2-yl)piperazine (III)
2-Chloro-4,6-disubstituted pyrimidines II were prepared by reaction of the
diazonium salts of 4,6-disubstituted pyrimidin-2-amines with concentrated
hydrochloric acid and ZnCl2 [43]. Compounds III were prepared according to the
known protocol [42] and the method was improved. To a stirred solution of piperazine
(45 mmol) and K2CO3 (16.5 mmol) in water (20 mL), chloropyrimidine II (18 mmol)
was added in small portions at 50-65 °C. The mixture was stirred for 1 h at 60-65 oC
o
and cooled to 35 C. The yellow solid (1,4-disubstituted piperazine byproduct), was
filtered off, and the filtrate was extracted three times with chloroform, dried over
Na2SO4, and concentrated to give III, which were used in the subsequent step without
further purification.
o
IIIa: yellow solid, yield 81%, mp 45-48 C; IIIb: yellow solid, yield 79%, mp
10

82-84 oC.
4.2.3
Preparation
of
5-chloro-1-phenyl-3-methyl/trifluoromethyl-1H-pyrazole-4-carbalde hyde (V)
2-Phenyl-5-methyl/trifluoromethyl-2,4-dihydro-3H-pyrazol-3-one IV were
prepared by refluxing ethyl acetoacetate or ethyl trifluoroacetoacetate and
phenylhydrazine in ethanol [37]. Intermediate pyrazole carbaldehydes V were
prepared according to the reference [39] and the method was improved. To a violently
stirred cold solution of DMF (23 mL), phosphorus oxychloride (64 mL) was added
dropwise at 0-10oC. The mixture was stirred at this temperature for 30 min; then
o
compound IV (0.1 mol) was added in portions. The mixture was heated at 90-100 C
for 3 ~ 4 h. After cooling down to room temperature, it was slowly poured into
ice-water (400 mL) and stirred for 10 min. For Va, the resulting solid was filtered off
and recrystallized from ethyl acetate/petroleum ether (60-90 °C) to give pure product;
for Vb, the mixture was extracted with ethyl acetate (3 × 30 mL). The combined
organic layer was washed with 10% NaHCO3 solution (2 × 20 mL) and brine (2 × 20
mL) successively, and dried over anhydrous Na2SO4. The solvent was removed under
reduced pressure, the residue was subjected to column chromatography using
petroleum ether and ethyl acetate (1:1) as solvents to give pure product.
o
Va: white crystals, yield 86%, mp 139-140 C (Lit. [44] mp 140-141oC); Vb:
yellow oil, yield 92% (Lit. [45] mp 34-36oC).
4.2.4
General
synthetic
procedures
for
4-(((5-chloro-1-phenyl-3-methyl/trifluoromethyl
-1H-
pyrazol-4-yl)methylene)amino)-5-(methyl/trifluoromethyl)-4H-1,2,4-triazole-3-th
iol (VII)
Compound VIa or VIb (10 mmol), pyrazole carbaldehyde V (10.5 mmol) and
acetic acid (10 drops) were mixed in absolute ethanol (25 mL). After being stirred and
refluxed for 4 ~ 5 h, the reaction mixture was cooled to room temperature. The
resulting solid was filtered off and recrystallized from ethanol to give Schiff base VII.
11

o
1
VIIa: colorless crystals, yield 83%, mp 224-226 C; H NMR (400 MHz,
DMSO-d6): δ 13.77 (s, 1H, SH), 10.18 (s, 1H, CH), 7.69-7.49 (m, 5H, Ph-H), 2.51 (s,
3H, CH3), 2.36 (s, 3H, CH3).
o
1
VIIb: white crystals, yield 72%, mp 206-207 C; H NMR (400 MHz, DMSO-d6):
δ 14.89 (s, 1H, SH), 10.12 (s, 1H, CH), 7.66-7.54 (m, 5H, Ph-H), 2.48 (s, 3H, CH3).
VIIc: colorless crystals, yield 87%, mp 202-204 oC; 1H NMR (400 MHz, CDCl3): δ
11.12 (s, 1H, SH), 10.91 (s, 1H, CH), 7.62-7.49 (m, 5H, Ph-H), 2.47 (s, 3H, CH3).
VIId: colorless crystals, yield 77%, mp 195-197 oC; 1H NMR (400 MHz, CDCl3) δ
11.52 (s, 1H, SH), 10.74 (s, 1H, CH), 7.60-7.50 (m, 5H, Ph-H).
4.2.5
General
synthetic
procedures
for
4-(((5-chloro-1-phenyl-3-methyl/trifluoromethyl-1H-
pyrazol-4-yl)methylene)amino)-2-((4-(substitutedbenzyl)piperazin-1-yl)methyl)-5
-(methyl/trifluoromethyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione (VIII)
Schiff base VII (1 mmol) and 37% formalin (1.5 mmol) were dissolved in ethanol
(15 mL), and the mixture was stirred at room temperature for 5 min. The solution of
4-(substituted benzyl)piperazine I (1 mmol) in ethanol (2 mL) was slowly added
dropwise. Then the reaction mixture was stirred for 2 ~ 3 h and left at room
temperature or placed in a refrigerator overnight. The resulting precipitate was filtered
off and recrystallized from ethanol to give novel 1,2,4-triazole thione VIII.
1-((4-Benzylpiperazin-1-yl)methyl)-4-(((5-chloro-3-methyl-1-phenyl-1H-pyrazo
l-4-yl)methylene)amino)-3-methyl-1H-1,2,4-triazole-5(4H)-thione (VIIIa) White
o
solid, yield 64%, mp 120-122 C; IR (KBr), ν (cm-1): 2935, 2805(C-H), 1612(C=N),
1
1596, 1537, 1500, 1454(Ar), 1167(C=S). H NMR (400 MHz, DMSO-d6): δ 9.96 (s,
1H, CH=N), 7.57-7.49 (m, 5H, Ph-H), 7.24-7.10 (m, 5H, Ph-H), 4.98 (s, 2H, NCH2N),
3.37 (s, 2H, PhCH2), 2.67 (bs, 4H, Piperazine-H), 2.33 (bs, 4H, Piperazine-H), 2.30 (s,
3H, Pyrazole-CH3). Anal. calcd for C26H29ClN8S: C, 59.93; H, 5.61; N, 21.50; found:
C, 59.65; H, 5.71; N 21.29.
4-(((5-Chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)methylene)amino)-1-((4-(2,4-
dichlorobenzyl)piperazin-1-yl)methyl)-3-methyl-1H-1,2,4-triazole-5(4H)-thione
12

(VIIIb) White solid, yield 67%, mp 147-149 oC; IR (KBr), ν (cm-1): 2940, 2813(C-H),
1615(C=N), 1593, 1531, 1504, 1455(Ar), 1165(C=S). 1H NMR (400 MHz,
DMSO-d6): δ 10.00 (s, 1H, CH=N), 7.63-7.57 (m, 6H, Ph-H), 7.46-7.38 (m, 2H,
Ph-H), 5.04 (s, 2H, NCH2N), 3.50 (s, 2H, ArCH2), 2.73 (bs, 4H, Piperazine-H), 2.52
13
(s, 3H, Triazole-CH3), 2.42 (bs, 4H, Piperazine-H), 2.39 (s, 3H, Pyrazole-CH3). C
NMR (101 MHz, CDCl3): δ 162.6 (C=S), 153.9 (triazole), 150.4 (pyrazole), 147.7
(C=N), 137.4 (Ph), 134.8 (Ph), 134.6 (Ph), 133.1 (Ph), 131.4 (Ph), 131.1 (pyrazole),
129.2 (Ph), 129.2 (Ph), 128.9 (Ph), 126.9 (Ph), 125.0 (Ph), 112.4 (pyrazole), 68.7
(CH2), 58.7 (CH2), 53.0 (piperazine-C), 50.5 (piperazine-C), 15.1 (CH3), 11.2 (CH3).
Anal. calcd for C26H27Cl3N8S: C, 52.93; H, 4.61; N, 18.99; found: C, 52.65; H, 4.49;
N, 19.11.
1-((4-Benzylpiperazin-1-yl)methyl)-4-(((5-chloro-3-methyl-1-phenyl-1H-pyrazo
l-4-yl)methylene)amino)-3-(trifluoromethyl)-1H-1,2,4-triazole-5(4H)-thione
o
(VIIIc) White crystals, yield 69%, mp 100-102 C; IR (KBr), ν (cm-1): 2940,
1
2878(C-H), 1608(C=N), 1535, 1489, 1456(Ar), 1312, 1197(C-F), 1154(C=S). H
NMR (400 MHz, DMSO-d6): δ 9.99 (s, 1H, CH=N), 7.65-7.54 (m, 5H, Ph-H),
7.32-7.23 (m, 5H, Ph-H), 5.17 (s, 2H, NCH2N), 3.46 (s, 2H, PhCH2), 2.77 (bs, 4H,
Piperazine-H), 2.47 (bs, 4H, Piperazine-H), 2.40 (s, 3H, Pyrazole-CH3). Anal. calcd
for C26H26ClF3N8S: C, 54.30; H, 4.56; N, 19.49; found: C, 54.38; H, 4.34; N, 19.21.
4-(((5-Chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)methylene)amino)-1-((4-(2,4-
dichlorobenzyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)-1H-1,2,4-triazole-5(4
H)-thione (VIIId) White solid, yield 74%, mp 128-130 oC; IR (KBr), ν (cm-1): 2941,
2818(C-H), 1607(C=N), 1535, 1502, 1488, 1457(Ar), 1314, 1196(C-F), 1163(C=S).
1H NMR (400 MHz, DMSO-d6): δ 9.98 (s, 1H, CH=N), 7.66-7.55 (m, 6H, Ph-H),
7.45 (d, J = 8.3 Hz, 1H, Ph-H), 7.39 (m, 1H, Ph-H), 5.18 (s, 2H, NCH2N), 3.51 (s, 2H,
ArCH2), 2.78 (bs, 4H, Piperazine-H), 2.50 (bs, 4H, Piperazine-H), 2.44 (s, 3H,
Pyrazole-CH3). 13C NMR (101 MHz, CDCl3): δ 165.1 (C=S), 155.2 (pyrazole), 151.0
(C=N), 138.8 (q, J = 47.7 Hz, triazole), 137.3 (Ph), 134.9 (Ph), 134.2 (Ph), 133.2 (Ph),
131.8 (Ph), 131.5 (pyrazole), 129.3 (Ph), 129.2 (Ph), 129.0 (Ph), 127.0 (Ph), 125.0
(Ph), 116.9 (q, J = 271.6 Hz, CF3), 111.9 (pyrazole), 69.9 (CH2), 58.6 (CH2), 52.9
13

(piperazine), 50.4 (piperazine), 15.0 (CH3). Anal. calcd for C26H24Cl3F3N8S: C, 48.49;
H, 3.76; N, 17.40; found: C, 48.55; H, 3.61; N, 17.30.
1-((4-Benzylpiperazin-1-yl)methyl)-4-(((5-chloro-1-phenyl-3-(trifluoromethyl)-
1H-pyrazol-4-yl)methylene)amino)-3-methyl-1H-1,2,4-triazole-5(4H)-thione
o
(VIIIe) White solid, yield 59%, mp 85-87 C; IR (KBr), ν (cm-1): 2938, 2813(C-H),
1596(C=N), 1533, 1492, 1452(Ar), 1315, 1191(C-F), 1150(C=S). 1H NMR (400 MHz,
DMSO-d6): δ 10.50 (s, 1H, CH=N), 7.72-7.63 (m, 5H, Ph-H), 7.29-7.23 (m, 5H,
Ph-H), 5.03 (s, 2H, NCH2N), 3.42 (s, 2H, PhCH2), 2.72 (bs, 4H, Piperazine-H), 2.36
(bs, 7H, Piperazine-H + Triazole-CH3). Anal. calcd for C26H26ClF3N8S: C, 54.30; H,
4.56; N, 19.49; found: C, 54.17; H, 4.62; N, 19.22.
4-(((5-Chloro-1-phenyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)methylene)amino)-
1-((4-(2,4-dichlorobenzyl)piperazin-1-yl)methyl)-3-methyl-1H-1,2,4-triazole-5(4H
o
)-thione (VIIIf) White crystals, yield 64%, mp 99-100 C; IR (KBr), ν (cm-1): 2941,
1
2825(C-H), 1593(C=N), 1532, 1493, 1458(Ar), 1298, 1176(C-F), 1143(C=S). H
NMR (400 MHz, CDCl3): δ 11.02 (s, 1H, CH=N), 7.68-7.41 (m, 5H, Ph-H), 7.39-7.35
(m, 2H, Ph-H), 7.18 (dd, J = 8.3, 2.1 Hz, 1H, Ph-H), 5.12 (s, 2H, NCH2N), 3.55 (s, 2H,
ArCH2), 2.87 (bs, 4H, Piperazine-H), 2.53 (bs, 4H, Piperazine-H), 2.45 (s, 3H,
Triazole-CH3). Anal. calcd for C26H24Cl3F3N8S: C, 48.49; H, 3.76; N, 17.40; found: C,
48.36; H, 3.78; N, 17.18.
4-(((5-Chloro-1-phenyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)methylene)amino)-
1-((4-(2,4-dichlorobenzyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)-1H-1,2,4-tri
o
azole-5(4H)-thione (VIIIg) White solid, yield 69%, mp 112-114 C; IR (KBr), ν
(cm-1): 2939, 2817(C-H), 1594(C=N), 1529, 1492, 1462(Ar), 1316, 1189(C-F),
1
1167(C=S). H NMR (400 MHz, DMSO-d6): δ 10.33 (s, 1H, CH=N), 7.75-7.72 (m,
2H, Ph-H), 7.66-7.63 (m, 3H, Ph-H), 7.56 (d, J = 1.9 Hz, 1H, Ph-H), 7.45 (d, J = 8.3
Hz, 1H, Ph-H), 7.38 (dd, J = 8.3, 1.9 Hz, 1H, Ph-H), 5.17 (s, 2H, NCH2N), 3.51 (s,
2H, ArCH2), 2.78 (bs, 4H, Piperazine-H), 2.44 (bs, 4H, Piperazine-H). 13C NMR (101
MHz, DMSO-d6): δ 164.7 (C=S), 140.0 (q, J = 39.2 Hz, triazole), 136.6 (Ph), 135.2
(Ph), 134.5 (Ph), 134.5 (pyrazole), 132.6 (Ph), 132.5 (Ph), 130.9 (pyrazole), 130.1
(Ph), 129.1 (Ph), 127.6 (Ph), 126.5 (Ph), 120.7 (q, J = 270.1 Hz, CF3), 117.2 (q, J =
14

272.7 Hz, CF3), 111.1 (pyrazole), 70.0 (CH2), 58.4 (CH2), 53.0 (piperazine-C), 50.0
(piperazine-C). Anal. calcd for C26H21Cl3F6N8S: C, 44.74; H, 3.03; N, 16.06; found: C,
44.52; H, 3.22; N, 15.98.
4.2.6
General
synthetic
procedures
for
4-(((5-Chloro-1-phenyl-3-methyl/trifluoromethyl-1H-
pyrazol-4-yl)methylene)amino)-2-((4-arylpiperazin-1-yl)methyl)-5-(methyl/triflu
oromethyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione (IX)
The procedure was similar to that of VIII, using 4-phenylpiperazine IIIc, or
4-(pyridin-2-yl)piperazine IIId, or 4-(4,6-disubstitutedpyrimidin-2-yl) piperazine
III(a-b) as amine material to give corresponding novel 1,2,4-triazole thione IX.
4-(((5-Chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)methylene)amino)-3-methyl-
1-((4-phenylpiperazin-1-yl)methyl)-1H-1,2,4-triazole-5(4H)-thione (IXa) White
o
solid, yield 72%, mp 152-154 C; IR (KBr), ν (cm-1): 2944, 2816(C-H), 1599(C=N),
1
1533, 1502, 1452(Ar), 1162(C=S). H NMR (400 MHz, CDCl3): δ 10.44 (s, 1H,
CH=N), 7.60-7.48 (m, 5H, Ph-H), 7.28 (t, J = 7.9 Hz, 2H, Ph-H), 6.94 (d, J = 8.3 Hz,
2H, Ph-H), 6.88 (t, J = 7.2 Hz, 1H, Ph-H), 5.22 (s, 2H, NCH2N), 3.25-3.22 (m, 4H,
Piperazine-H), 3.04-3.02 (m, 4H, Piperazine-H), 2.61 (s, 3H, Triazole-CH3), 2.47 (s,
3H, Pyrazole-CH3). Anal. calcd for C25H27ClN8S: C, 59.22; H, 5.37; N, 22.10; found:
C, 58.98; H, 5.29; N, 22.00.
4-(((5-Chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)methylene)amino)-3-methyl-
1-((4-(pyridin-2-yl)piperazin-1-yl)methyl)-1H-1,2,4-triazole-5(4H)-thione (IXb)
o
White solid, yield 70%, mp 111-113 C; IR (KBr), ν (cm-1): 2945, 2842(C-H),
1
1594(C=N), 1534, 1481(Ar), 1161(C=S). H NMR (400 MHz, DMSO-d6): δ 9.99 (s,
1H, CH=N), 8.07 (d, J = 3.2 Hz, 1H, Pyridine-H), 7.62-7.57 (m, 5H, Ph-H), 7.52-7.46
(m, 1H, Pyridine-H), 6.80 (d, J = 8.6 Hz, 1H, Pyridine-H), 6.60 (dd, J = 6.9, 5.0 Hz,
1H, Pyridine-H), 5.10 (s, 2H, NCH2N), 3.52-3.43 (m, 4H, Piperazine-H), 2.87-2.73
(m, 4H, Piperazine-H), 2.50 (s, 3H, Triazole-CH3), 2.36 (s, 3H, Pyrazole-CH3). Anal.
calcd for C24H26ClN9S: C, 56.74; H, 5.16; N, 24.81; found: C, 56.79; H, 5.25; N,
24.85.
15

4-(((5-Chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)methylene)amino)-3-methyl-
1-((4-(4-methylpyrimidin-2-yl)piperazin-1-yl)methyl)-1H-1,2,4-triazole-5(4H)-thi
o
one (IXc) White solid, yield 80%, mp 155-157 C; IR (KBr), ν (cm-1): 2949,
1
2858(C-H), 1581(C=N), 1555, 1532, 1500, 1449(Ar), 1156(C=S). H NMR (400
MHz, DMSO-d6): δ 9.99 (s, 1H, CH=N), 8.17 (d, J = 4.8 Hz, 1H, Pyrimidine-H),
7.64-7.54 (m, 5H, Ph-H), 6.48 (d, J = 4.8 Hz, 1H, Pyrimidine-H), 5.10 (s, 2H,
NCH2N), 3.75-3.73 (m, 4H, Piperazine-H), 2.76-2.73 (m, 4H, Piperazine-H), 2.50 (s,
3H, Triazole-CH3), 2.36 (s, 3H, Pyrazole-CH3), 2.24 (s, 3H, Pyrimidine-CH3). Anal.
calcd for C24H27ClN10S: C, 55.11; H, 5.20; N, 26.78; found: C, 54.99; H, 5.11; N,
26.53.
4-(((5-Chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)methylene)amino)-1-((4-(4,6-
dimethylpyrimidin-2-yl)piperazin-1-yl)methyl)-3-methyl-1H-1,2,4-triazole-5(4H)
o
-thione (IXd) White solid, yield 85%, mp 152-154 C; IR (KBr), ν (cm-1): 2947,
1
2847(C-H), 1577(C=N), 1538, 1483, 1449(Ar), 1161(C=S). H NMR (400 MHz,
DMSO-d6): δ 9.99 (s, 1H, CH=N), 7.70-7.50 (m, 5H, Ph-H), 6.36 (s, 1H,
Pyrimidine-H), 5.09 (s, 2H, NCH2N), 3.74 (bs, 4H, Piperazine-H), 2.73 (bs, 4H,
Piperazine-H), 2.50 (s, 3H, Triazole-CH3), 2.35 (s, 3H, Pyrazole-CH3), 2.19 (s, 6H,
13
Pyrimidine-CH3). C NMR (101 MHz, CDCl3): δ 167.0 (pyrimidine), 162.6 (C=S),
161.6 (pyrimidine), 153.9 (triazole), 150.4 (pyrazole), 147.6 (C=N), 137.4 (Ph), 131.1
(pyrazole), 129.2 (Ph), 128.9 (Ph), 125.0 (Ph), 112.4 (pyrazole), 108.8 (pyrimidine),
68.9 (CH2), 50.7 (piperazine-C), 43.5 (piperazine-C), 24.1 (CH3), 15.1 (CH3), 11.1
(CH3). Anal. calcd for C25H29ClN10S: C, 55.91; H, 5.44; N, 26.08; found: C, 55.61; H,
5.51; N, 26.08.
4-(((5-Chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)methylene)amino)-1-((4-phe
nylpiperazin-1-yl)methyl)-3-(trifluoromethyl)-1H-1,2,4-triazole-5(4H)-thione
o
(IXe) White solid, yield 71%, mp 123-125 C; IR (KBr), ν (cm-1): 2952, 2828(C-H),
1604(C=N), 1530, 1491, 1454(Ar), 1316, 1198(C-F), 1168(C=S). 1H NMR (400 MHz,
DMSO-d6): δ 9.99 (s, 1H, CH=N), 7.65-7.50 (m, 5H, Ph-H), 7.19 (t, J = 7.3 Hz, 2H,
Ph-H), 6.92 (d, J = 7.8 Hz, 2H, Ph-H), 6.77 (t, J = 7.0 Hz, 1H, Ph-H), 5.25 (s, 2H,
NCH2N), 3.14 (bs, 4H, Piperazine-H), 2.91 (bs, 4H, Piperazine-H), 2.48 (s, 3H,
16

Pyrazole-CH3). Anal. calcd for C25H24ClF3N8S: C, 53.52; H, 4.31; N, 19.97; found: C,
53.36; H, 4.22; N, 19.88.
4-(((5-Chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)methylene)amino)-1-((4-(pyr
idin-2-yl)piperazin-1-yl)methyl)-3-(trifluoromethyl)-1H-1,2,4-triazole-5(4H)-thio
ne (IXf) White solid, yield 73%, mp 114-116 oC; IR (KBr), ν (cm-1): 2949, 2835(C-H),
1597(C=N), 1569, 1533, 1483(Ar), 1313, 1196(C-F), 1170(C=S). 1H NMR (400 MHz,
DMSO-d6): δ 9.97 (s, 1H, CH=N), 8.08 (dd, J = 4.8, 1.8 Hz, 1H, Pyridine-H),
7.65-7.59 (m, 6H, Ph-H + Pyridine-H), 6.82 (d, J = 8.6 Hz, 1H, Pyridine-H), 6.60 (dd,
J = 7.0, 5.0 Hz, 1H, Pyridine-H), 5.24 (s, 2H, NCH2N), 3.52-3.49 (m, 4H,
Piperazine-H), 2.86-2.83 (m, 4H, Piperazine-H), 2.47 (s, 3H, Pyrazole-CH3). Anal.
calcd for C24H23ClF3N9S: C, 52.93; H, 4.61; N, 18.99; found: C, 52.57; H, 4.69; N,
18.65.
4-(((5-Chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)methylene)amino)-1-((4-(4-
methylpyrimidin-2-yl)piperazin-1-yl)methyl)-3-(trifluoromethyl)-1H-1,2,4-triazol
o
e-5(4H)-thione (IXg) White solid, yield 81%, mp 153-155 C; IR (KBr), ν (cm-1):
2935, 2829(C-H), 1599(C=N), 1569, 1531, 1487, 1448(Ar), 1317, 1199(C-F),
1
1155(C=S). H NMR (400 MHz, DMSO-d6): δ 9.96 (s, 1H, CH=N), 8.17 (s, J = 3.4
Hz, 1H, Pyrimidine-H), 7.65-7.50 (m, 5H, Ph-H), 6.47 (d, J = 3.7 Hz, 1H,
Pyrimidine-H), 5.23 (s, 2H, NCH2N), 3.76 (bs, 4H, Piperazine-H), 2.80 (bs, 4H,
Piperazine-H), 2.46 (s, 3H, Pyrazole-CH3), 2.24 (s, 3H, Pyrimidine-CH3). Anal. calcd
for C24H24ClF3N10S: C, 49.96; H, 4.19; N, 24.27; found: C, 49.71; H, 4.32; N, 24.07.
4-(((5-Chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)methylene)amino)-1-((4-(4,6-
dimethylpyrimidin-2-yl)piperazin-1-yl)methyl)-3-(trifluoromethyl)-1H-1,2,4-tria
zole-5(4H)-thione (IXh) White solid, yield 84%, mp 168-170 oC; IR (KBr), ν (cm-1):
2937, 2831(C-H), 1600(C=N), 1582, 1536, 1493, 1448(Ar), 1315, 1197(C-F),
1152(C=S). 1H NMR (400 MHz, DMSO-d6): δ 9.96 (s, 1H, CH=N), 7.70-7.50 (m, 5H,
Ph-H), 6.36 (s, 1H, Pyrimidine-H), 5.23 (s, 2H, NCH2N), 3.76 (bs, 4H, Piperazine-H),
13
2.79 (bs, 4H, Piperazine-H), 2.46 (s, 3H, Me), 2.19 (s, 6H, Pyrimidine-CH3). C
NMR (101 MHz, CDCl3): δ 167.1 (pyrimidine), 165.0 (C=S), 161.5 (pyrimidine),
159.4 (m, triazole), 155.3 (pyrazole), 151.0 (C=N), 137.3 (Ph), 131.9 (pyrazole),
17

129.3 (Ph), 129.0 (Ph), 125.0 (Ph), 116.9 (q, J = 271.9 Hz, CF3), 111.9 (pyrazole),
109.0 (pyrimidine), 70.1 (CH2), 50.5 (piperazine-C), 43.4 (piperazine-C), 24.1 (CH3),
15.0 (CH3). Anal. calcd for C25H26ClF3N10S: C, 50.80; H, 4.43; N, 23.70; found: C,
50.52; H, 4.54; N, 23.34.
4-(((5-Chloro-1-phenyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)methylene)amino)-
3-methyl-1-((4-phenylpiperazin-1-yl)methyl)-1H-1,2,4-triazole-5(4H)-thione (IXi)
o
White solid, yield 73%, mp 139-141 C; IR (KBr), ν (cm-1): 2944, 2825(C-H),
1598(C=N), 1528, 1494, 1451(Ar), 1322, 1182(C-F), 1146(C=S). 1H NMR (400 MHz,
DMSO-d6): δ 10.50 (s, 1H, CH=N), 7.75-7.72 (m, 2H, Ph-H), 7.67-7.64 (m, 3H,
Ph-H), 7.19 (t, J = 7.8 Hz, 2H, Ph-H), 6.91 (d, J = 8.3 Hz, 2H, Ph-H), 6.77 (t, J = 7.2
Hz, 1H, Ph-H), 5.12 (s, 2H, NCH2N), 3.13 (bs, 4H, Piperazine-H), 2.87 (bs, 4H,
Piperazine-H), 2.38 (s, 3H, Triazole-CH3). Anal. calcd for C25H24ClF3N8S: C, 53.52;
H, 4.31; N, 19.97; found: C, 53.37; H, 4.34; N, 19.83.
4-(((5-Chloro-1-phenyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)methylene)amino)-
3-methyl-1-((4-(pyridin-2-yl)piperazin-1-yl)methyl)-1H-1,2,4-triazole-5(4H)-thio
o
ne (IXj) White solid, yield 75%, mp 145-147 C; IR (KBr), ν (cm-1): 2939,
2833(C-H), 1595(C=N), 1532, 1488(Ar), 1300(C-F), 1174(C=S). 1H NMR (400 MHz,
DMSO-d6): δ 10.49 (s, 1H, CH=N), 8.07 (d, J = 3.4 Hz, 1H, Pyridine-H), 7.73-7.71
(m, 2H, Ph-H), 7.65-7.63 (m, 3H, Ph-H), 7.56-7.40 (m, 1H, Pyridine-H), 6.80 (d, J =
8.6 Hz, 1H, Pyridine-H), 6.60 (dd, J = 6.8, 5.1 Hz, 1H, Pyridine-H), 5.10 (s, 2H,
NCH2N), 3.49 (bs, 4H, Piperazine-H), 2.80 (bs, 4H, Piperazine-H), 2.35 (s, 3H,
Triazole-CH3). Anal. calcd for C24H23ClF3N9S: C, 51.29; H, 4.12; N, 22.43; found: C,
51.25; H, 4.09; N, 22.26.
4-(((5-Chloro-1-phenyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)methylene)amino)-
3-methyl-1-((4-(4-methylpyrimidin-2-yl)piperazin-1-yl)methyl)-1H-1,2,4-triazole-
5(4H)-thione (IXk) White solid, yield 75%, mp 203-204 oC; IR (KBr), ν (cm-1): 2937,
2861(C-H), 1577(C=N), 1530, 1492, 1450(Ar), 1296(C-F), 1177(C=S). 1H NMR (400
MHz, CDCl3): δ 11.00 (s, 1H, CH=N), 8.13 (d, J = 4.9 Hz, 1H, Pyri midine-H), 7.56
(m, 5H, Ph-H), 6.34 (d, J = 5.0 Hz, 1H, Pyrimidine-H), 5.16 (s, 2H, NCH2N),
3.92-3.80 (m, 4H, Piperazine-H), 2.92-2.82 (m, 4H, Piperazine-H), 2.42 (s, 3H,
18

Triazole-CH3), 2.30 (s, 3H, Pyrimidine-CH3). Anal. calcd for C24H24ClF3N10S: C,
49.96; H, 4.19; N, 24.27; found: C, 49.65; H, 4.23; N, 24.18.
4-(((5-Chloro-1-phenyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)methylene)amino)-
1-((4-(4,6-dimethylpyrimidin-2-yl)piperazin-1-yl)methyl)-3-methyl-1H-1,2,4-triaz
o
ole-5(4H)-thione (IXl) White solid, yield 78%, mp 191-193 C; IR (KBr), ν (cm-1):
2938, 2832(C-H), 1580(C=N), 1533, 1494, 1451(Ar), 1295, 1174(C-F), 1161(C=S).
1H NMR (400 MHz, DMSO-d6): δ 10.99 (s, 1H, CH=N), 7.57-7.53 (m, 5H, Ph-H),
6.23 (s, 1H, Pyrimidine-H), 5.16 (s, 2H, NCH2N), 3.89-3.86 (bs, 4H, Piperazine-H),
2.88-2.85 (bs, 4H, Piperazine-H), 2.41 (s, 3H, Triazole-CH3), 2.25 (s, 6H,
13
Pyrimidine-CH3). C NMR (101 MHz, CDCl3): δ 167.1 (pyrimidine), 162.6 (C=S),
161.6 (pyrimidine), 148.3 (triazole), 147.5 (C=N), 141.1 (q, J = 39.6 Hz, pyrazole),
136.7 (Ph), 132.1 (pyrazole), 130.0 (Ph), 129.5 (Ph), 125.4 (Ph), 120.4 (q, J = 270.3
Hz, CF3), 112.7 (pyrazole), 108.9 (pyrimidine), 68.7 (CH2), 50.7 (piperazine-C), 43.5
(piperazine-C), 24.1 (CH3), 11.0 (CH3). Anal. calcd for C25H26ClF3N10S: C, 50.80; H,
4.43; N, 23.70; found: C, 50.58; H, 4.54; N, 23.49.
4-(((5-Chloro-1-phenyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)methylene)amino)-
1-((4-phenylpiperazin-1-yl)methyl)-3-(trifluoromethyl)-1H-1,2,4-triazole-5(4H)-t
o
hione (IXm) White solid, yield 76%, mp 148-150 C; IR (KBr), ν (cm-1): 2952,
2829(C-H), 1601(C=N), 1579, 1528, 1495, 1456(Ar), 1319, 1197(C-F), 1176(C=S).
1H NMR (400 MHz, DMSO-d6): δ 10.33 (s, 1H, CH=N), 7.73 (d, J = 3.6 Hz, 2H,
Ph-H), 7.64 (d, J = 3.4 Hz, 3H, Ph-H), 7.19 (t, J = 7.5 Hz, 2H, Ph-H), 6.92 (d, J = 7.9
Hz, 2H, Ph-H), 6.76 (t, J = 7.0 Hz, 1H, Ph-H), 5.25 (s, 2H, NCH2N), 3.13 (bs, 4H,
Piperazine-H), 2.92 (bs, 4H, Piperazine-H). Anal. calcd for C25H21ClF6N8S: C, 48.82;
H, 3.44; N, 18.22; found: C, 48.81; H, 3.65; N, 18.00.
4-(((5-Chloro-1-phenyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)methylene)amino)-
1-((4-(pyridin-2-yl)piperazin-1-yl)methyl)-3-(trifluoromethyl)-1H-1,2,4-triazole-5
(4H)-thione (IXn) White solid, yield 79%, mp 125-127 oC; IR (KBr), ν (cm-1): 2948,
1
2847(C-H), 1596(C=N), 1567, 1531, 1487(Ar), 1317, 1186(C-F), 1164(C=S). H
NMR (400 MHz, DMSO-d6): δ 10.31 (s, 1H, CH=N), 8.08 (dd, J = 4.8, 1.6 Hz, 1H,
Pyridine-H), 7.74 (dd, J = 6.5, 3.0 Hz, 2H, Ph-H), 7.65-7.63 (m, 3H, Ph-H), 7.51-7.47
19

(m, 1H, Pyridine-H), 6.82 (d, J = 8.6 Hz, 1H, Pyridine-H), 6.60 (dd, J = 7.0, 5.0 Hz,
1H, Pyridine-H), 5.24 (s, 2H, NCH2N), 3.51-3.49 (m, 4H, Piperazine-H), 2.86-2.84
(m, 4H, Piperazine-H). Anal. calcd for C24H20ClF6N9S: C, 46.80; H, 3.27; N, 20.46;
found: C, 46.59; H, 3.41; N, 20.25.
4-(((5-Chloro-1-phenyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)methylene)amino)-
1-((4-(4-methylpyrimidin-2-yl)piperazin-1-yl)methyl)-3-(trifluoromethyl)-1H-1,2,
o
4-triazole-5(4H)-thione (IXo) White solid, yield 73%, mp 108-110 C; IR (KBr), ν
(cm-1): 2936, 2828(C-H), 1611(C=N), 1575, 1531, 1487, 1449(Ar), 1299, 1212(C-F),
1169(C=S). 1H NMR (400 MHz, DMSO-d6), δ: 10.30 (s, 1H, CH=N), 8.17 (d, J = 4.9
Hz, 1H, Pyrimidine-H), 7.75-7.72 (m, 2H, Ph-H), 7.65-7.63 (m, 3H, Ph-H), 6.47 (d, J
= 4.9 Hz, 1H, Pyrimidine-H), 5.23 (s, 2H, NCH2N), 3.76 (bs, 4H, Piperazine-H),
2.82-2.79 (m, 4H, Piperazine-H), 2.24 (s, 3H, Pyrimidine-CH3). Anal. calcd for
C24H21ClF6N10S: C, 45.68; H, 3.35; N, 22.20; found: C, 45.41; H, 3.36; N, 22.12.
4-(((5-Chloro-1-phenyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)methylene)amino)-
1-((4-(4,6-dimethylpyrimidin-2-yl)piperazin-1-yl)methyl)-3-(trifluoromethyl)-1H-
1,2,4-triazole-5(4H)-thione (IXp) White solid, yield 83%, mp 120-122 oC; IR (KBr),
ν (cm-1): 2822(C-H), 1614(C=N), 1576, 1532, 1492, 1449(Ar), 1303, 1181(C-F),
1
1165(C=S). H NMR (400 MHz, DMSO-d6): δ 10.30 (s, 1H, CH=N), 7.75-7.72 (m,
2H, Ph-H), 7.65-7.63 (m, 3H, Ph-H), 6.35 (s, 1H, Pyrimidine-H), 5.23 (s, 2H,
NCH2N), 3.76 (bs, 4H, Piperazine-H), 2.79 (bs, 4H, Piperazine-H), 2.19 (s, 6H,
13
Pyrimidine-CH3). C NMR (101 MHz, CDCl3): δ 167.1 (pyrimidine), 165.0 (C=S),
161.6 (pyrimidine), 150.4 (C=N), 141.9 (q, J = 39.5 Hz, triazole), 138.9 (d, J = 42.4
Hz, pyrazole), 136.6 (Ph), 132.4 (pyrazole), 130.1 (Ph), 129.5 (Ph), 125.4 (Ph), 120.1
(q, J = 270.6 Hz, CF3),116.7 (d, J = 271.9 Hz, CF3), 111.5 (pyrazole), 109.1
(pyrimidine), 70.1 (CH2), 50.5 (piperazine-C), 43.5 (piperazine-C), 24.1 (CH3). Anal.
calcd for C25H23ClF6N10S: C, 46.55; H, 3.59; N, 21.71; found: C, 46.54; H, 3.55; N,
21.54.
4.2.7
General
synthetic
procedures
for
1,1’-((piperazin-1,4-diyl)bis(methylene))bis(4-
20

((5-chloro-1-phenyl-3-methyl/trifluoromethyl-1H-pyrazol-4-yl)methylene)amino-
3-methyl/trifluoromethyl-1H-1,2,4-triazole-5(4H)-thione) (X)
Schiff base VII (1.8 mmmol) and 37% formalin (2.5 mmol) were dissolved in
ethanol (30 mL), and the mixture was stirred at room temperature for 5 min. A
solution of piperazine (0.9 mmol) in ethanol (2 mL) was slowly added dropwise. Then
the reaction mixture was stirred for 1 ~ 2 h at room temperature. The resulting
precipitate was filtered off and recrystallized from ethanol to give novel
bis(1,2,4-triazole thione) X.
1,1’-(Piperazine-1,4-diylbis(methylene))bis(4-(((5-chloro-3-methyl-1-phenyl-1
H-pyrazol-4-yl)methylene)amino)-3-methyl-1H-1,2,4-triazole-5(4H)-thione) (Xa)
White solid, yield 83%, mp 224-226oC(Dec.); IR (KBr), ν (cm-1): 2936, 2815(C-H),
1597(C=N), 1539, 1504, 1485, 1451(Ar), 1169(C=S). 1H NMR (400 MHz, CDCl3): δ
10.39 (s, 2H, CH=N), 7.60-7.40 (m, 10H, Ph-H), 5.08 (s, 4H, NCH2N), 2.86 (s, 8H,
Piperazine-H), 2.57 (s, 6H, Pyrazole-CH3), 2.41 (s, 6H, Triazole-CH3). Anal. calcd for
C34H36Cl2N14S2: C, 52.64; H, 4.68; N, 25.28; found: C, 52.81; H, 4.65; N, 25.13.
1,1’-(Piperazine-1,4-diylbis(methylene))bis(4-(((5-chloro-3-methyl-1-phenyl-1
H-pyrazol-4-yl)methylene)amino)-3-(trifluoromethyl)-1H-1,2,4-triazole-5(4H)-thi
one) (Xb) White solid, yield 87%, mp 185-187oC(Dec.); IR (KBr), ν (cm-1): 2940,
2857(C-H), 1599(C=N), 1533, 1504, 1485, 1460(Ar), 1315, 1189(C-F), 1171(C=S).
1H NMR (400 MHz, CDCl3): δ 10.35 (s, 2H, CH=N), 7.60-7.40 (m, 10H, Ph-H), 5.18
(s, 4H, NCH2N), 2.89 (s, 8H, Piperazine-H), 2.56 (s, 6H, Pyrazole-CH3). 13C NMR
(101 MHz, CDCl3): δ 165.0 (C=S), 155.3 (pyrazole), 151.0 (C=N), 138.8 (q, J = 42.0
Hz, triazole), 137.3 (Ph), 131.9 (pyrazole), 129.3(Ph), 129.0(Ph), 125.0 (Ph), 116.9 (q,
J = 271.6 Hz, CF3), 111.9 (pyrazole), 69.8 (CH2), 50.3 (piperazine), 15.0 (CH3). Anal.
calcd for C34H30Cl2F6N14S2: C, 46.21; H, 3.42; N, 22.19; found: C, 46.38; H, 3.61; N,
21.99.
1,1’-(Piperazine-1,4-diylbis(methylene))bis(4-(((5-chloro-1-phenyl-3-(trifluoro
methyl)-1H-pyrazol-4-yl)methylene)amino)-3-methyl-1H-1,2,4-triazole-5(4H)-thi
one) (Xc) White solid, yield 89%, mp 213-215oC(Dec.); IR (KBr), ν (cm-1): 2939,
2828(C-H), 1594(C=N), 1530, 1494, 1453(Ar), 1326(C-F), 1175(C=S). 1H NMR (400
21

MHz, CDCl3): δ 10.99 (s, 2H, CH=N), 7.60-7.50 (m, 10H, Ph-H), 5.06 (s, 4H,
NCH2N), 2.85 (s, 8H, Piperazine-H), 2.42 (s, 6H, Triazole-CH3). Anal. calcd for
C34H30Cl2F6N14S2: C, 46.21; H, 3.42; N, 22.19; found: C, 46.08; H, 3.39; N, 22.29.
1,1’-(Piperazine-1,4-diylbis(methylene))bis(4-(((5-chloro-1-phenyl-3-(trifluoro
methyl)-1H-pyrazol-4-yl)methylene)amino)-3-(trifluoromethyl)-1H-1,2,4-triazole
-5(4H)-thione) (Xd) White solid, yield 84%, mp 197-198oC(Dec.); IR (KBr), ν (cm-1):
2945, 2851(C-H), 1597(C=N), 1535, 1493, 1462(Ar), 1322, 1186(C-F), 1164(C=S).
1H NMR (400 MHz, CDCl3): δ 10.82 (s, 2H, CH=N), 7.58-7.50 (m, 10H, Ph-H), 5.17
13
(s, 4H, NCH2N), 2.88 (s, 8H, Piperazine-H). C NMR (101 MHz, CDCl3): δ 164.9
(C=S), 150.4 (C=N), 141.9 (q, J = 39.8 Hz, triazole), 139.0 (q, J = 41.8 Hz, pyrazole),
136.6 (Ph), 132.5 (pyrazole), 130.1 (Ph), 129.5 (Ph), 125.4 (Ph), 120.1 (q, J = 270.7
Hz, CF3), 116.7 (q, J = 271.3 Hz, CF3), 111.4 (pyrazole), 69.7 (CH2), 50.3
(piperazine). Anal. calcd for C34H24Cl2F12N14S2: C, 41.18; H, 2.44; N, 19.77; found: C,
40.92; H, 2.48; N, 19.71.
4.3 Biological activity tests
4.3.1 Herbicidal activity test
The in vivo herbicidal activities were determined by the inhibition tests of the
root-growth of rape (Brassica campestris) and the seedling-growth of barnyardgrass
(Echinochloa crusgalli) according to the reported protocol [46]. The commercial
herbicide Chlorsulfuron was used as the control.
4.3.2 Fungicidal activity test
The in vitro fungicidal activities against Alternaria solani Sorauer, Gibberella
sanbinetti, Fusarium omysporum, Cercospora arachidicola, Physalospora piricola
and Rhizoctonia cerealis were evaluated via the mycelium growth rate test according
to the reported protocol [47]. The commercial fungicides Triadimefon, Carbendazim
and Chlorothalonil were used as controls.
The in vivo fungicidal activities against Puccinia sorghi Schw. was tested
according to the reported method [48], and the commercial fungicides Azoxystrobin
22

and Triadimefon were used as controls.
Acknowledgements
This work was supported by the National Natural Science Foundation of China (No.
21372133) and “111” Project of Ministry of Education of China (No. B06005). We
thank Dr. Yong-Hong Li of the Biological Assay Center, Nankai University, for
herbicidal tests of compounds.
Supplementary data
All the supplementary data associated with this article including 1H NMR and IR
spectra for compounds VIII, IX and X, 13C NMR spectra for representative
compounds and herbicidal and in vitro fungicidal activities for some of the
compounds can be found in the online version.
23

References
[1] L.F. Lee, F.M. Schleppnik, R.W. Schneider, D.H. Campbell, J. Heterocyclic Chem.
27 (1990) 243-245.
[2] J. Zhang, C.F. Jin, Y.J. Zhang, Chin. J. Org. Chem. 34 (2014) 662-680.
[3] M.F. Abdel-Megeed, B.E. Badr, M.M. Azaam, G.A. El-Hiti, Phosphorus Sulfur
Silicon Relat. Elem. 187 (2012) 1462-1468.
[4] J.-C. Yang, Q. Wu, Y.-P. Liu, C.-L. Liu, Agrochemicals 50(4)(2011) 289-295.
[5] W. Zhu, J. Wang, S. Wang, Z. Gu, J.L. Aceña, K. Izawa, H. Liu, V.A. Soloshonok,
J. Fluorine Chem. 167 (2014) 37-54.
[6] K.L. Kirk, J. Fluorine Chem. 127 (2006) 1013-1029.
[7] M.J. Corr, D. O’Hagan, J. Fluorine Chem. 155 (2013) 72-77.
[8] J. Nieschalk, D. O’Hagan, J. Fluorine Chem. 91 (1998) 159-163.
[9] J. Wang, M. Sánchez-Roselló, J.L. Aceña, C. del Pozo, A.E. Sorochinsky, S.
Fustero, V.A. Soloshonok, H. Liu, Chem. Rev. 114 (2014) 2432-2506.
[10] A. Guan, C. Liu, G. Huang, H. Li, S. Hao, Y. Xu, Y. Xie, Z. Li, J. Fluorine Chem.
160 (2014) 82-87.
[11] M. Tomizawa, J.E. Casida, J. Agric. Food Chem. 59 (2011) 2883-2886.
[12]
M.A. Matocha, L.J. Krutz,
K.N. Reddy, S.A.Senseman, M.A. Locke,
R.W. Steinriede, Jr., E.W. Palmer, J. Agric. Food Chem. 54 (2006) 5498-5502.
[13] A.L. Blobaum, Md.J. Uddin, A.S. Felts, B.C. Crews, C.A. Rouzer, L.J. Marnett,
ACS Med. Chem. Lett. 4 (2013) 486-490.
[14]
G.-H. Kuo, T.
Rano, P. Pelton, K.T. Demarest, A.C.
Gibbs, W.V. Murray, B.P. Damiano, M.A. Connelly, J. Med. Chem. 52 (2009)
1768-1772.
[15] G. Szabó, J. Fischer, Á. Kis-Varga, K. Gyires, J. Med. Chem. 51 (2008) 142-147.
[16] K. Kaur, V. Kumar, G.K. Gupta, J. Fluorine Chem. 178 (2015) 306-326.
[17] X.-M. Xu, F.-F Zhao, X. Zheng, Y.-M. Liu, X. Yao, X. Luo, J. Huang, Z.-T.
Zheng, M.-H. Niu, R.-F. Liao, J. Univ. South China (Med. Ed.) 37(2) (2009) 129-132.
[18] S.M. Wilhelm, L. Adnane, P. Newell, A. Villanueva, J.M. Llovet, M. Lynch, Mol.
Cancer Ther. 7 (2008) 3129-3140.
24

[19] M. Tohnishi, H. Nakao, T. Furuya, A. Seo, H. Kodama, K. Tsubata, J. Pestic. Sci.
30 (2005) 354-360.
[20] S.N. Pandeya, D. Sriram, P. Yogeeswari, S. Ananthan, Chemotherapy 47 (2001)
266-269.
[21] C. Chen, X.Y. Zheng, P.E. Zhu, H.Z. Guo, Acta Pharm. Sinica 2 (1982) 112-117.
[22] S. Liang, C.-M. Liu, Y.-S. Jin, Q.-Q. He, Chin. J. Med. Chem. 14 (2004) 71-75.
[23] A.S. Aboraia, H.M. Abdel-Rahman, N.M. Mahfouz, M.A. El-Gendy, Bioorg.
Med. Chem. 14 (2006) 1236-1246.
[24] A. Tanitame, Y. Oyamada, K. Ofuji, M. Fujimoto, N. Iwai, Y. Hiyama, K.
Suzuki, H. Ito, H. Terauchi, M. Kawasaki, K. Nagai, M. Wachi, J. Yamagishi, J. Med.
Chem. 47 (2004) 3693-3696.
[25] G.P. Ouyang, X.J. Cai, Z. Chen, B.A. Song, P.S. Bhadury, S. Yang, L.H. Jin, W.
Xue, D.Y. Hu, S. Zeng, J. Agric. Food Chem. 56 (2008) 10160-10167.
[26] M.R. Zhang, J. Liu, X.W. Zhang, Y.P. Zhou, Y. Chen, Y. Wang, L.F. Xu, Chin.
Pharm. J. 50 (2015) 194-200.
[27] H. Dai, Y.Q. Li, D. Du, X. Qin, X. Zhang, H.B. Yu, J.X. Fang, J. Agric. Food
Chem. 56 (2008) 10805-10810.
[28] J. Cao, Y.-B. Zhang, Fine Chem. Interm. 37(6) (2007) 1-4.
[29] Y.-C. Lang, Y.-L. Bai, Modern Agrochem. 5(5) (2006) 6-12.
[30] B.-L. Wang, Y.-X. Shi, Y. Ma, X.-H. Liu, Y.-H. Li, H.-B. Song, B.-J. Li, Z.-M.
Li, J. Agric. Food Chem. 58 (2010) 5515-5522.
[31] B.-L. Wang, X.-H. Liu, X.-L. Zhang, J.-F. Zhang, H.-B. Song, Z.-M. Li, Chem.
Biol. Drug Des. 78 (2011) 8 42-49.
[32] D.O. Berbasov, I.D. Ojemaye, V.A. Soloshonok, J. Fluorine Chem. 125 (2004)
603-607.
[33] V.A. Soloshonok, D.O. Berbasov, J. Fluorine Chem. 125 (2004) 1757-1763.
[34] H. Ohkura, D.O. Berbasov, V.A. Soloshonok, Tetrahedron 59 (2003) 1647-1656.
[35] H. Ohkura, D.O. Berbasov, V.A. Soloshonok, Tetrahedron Lett. 44 (2003)
2417-2420.
[36] D.O. Berbasov, V.A. Soloshonok, Synthesis (13) (2003) 2005-2010.
25

[37] N.N. Dykhanov, Med. Prom. SSSR 1 (1961) 42-45.
[38] B.-L. Wang, Q.-N. Li, Y.-Z. Zhan, L.-X. Xiong, S.-J. Yu, Z.-M. Li, Phosphorus
Sulfur Silicon Relat. Elem.189 (2014) 483-491.
[39] B.A. Porai-Koshits, I.Ya. Kvitko, E.A. Shutkova, Pharm. Chem. J. 4 (1970)
138-143.
[40] Z. Liu, X. Zhang, W. Lu, Jiangsu J. Agr. Sci. 16 (2000) 185-190.
[41] T. George, D.V. Mehta, R. Tahilramani, J. David, P. K. Talwalker, J. Med. Chem.
14 (1971) 335-338.
[42] P. Zlatoidsky´, T. Maliar, Eur. J. Med. Chem. 31 (1996) 669-674.
[43] R.R. Adams, F.C. Whitmore, J. Am. Chem. Soc. 67 (1945) 735-738.
[44] M. Nayak, H. Batchu, S. Batra, Tetrahedron Lett. 53 (2012) 4206-4208.
[45] B. Palka, A.D. Capua, M. Anzini, G. Vilkauskaité, A. Šačkus, W. Holzer,
Beilstein J. Org. Chem. 10 (2014) 1759-1764.
[46] B.-L. Wang, R.G. Duggleby, Z.-M. Li, J.-G. Wang, Y.-H. Li, S.-H. Wang, H.-B.
Song, Pest Manag. Sci. 61 (2005) 407-412.
[47] Z. Liu, G. Yang, X. Qin, J. Chem. Technol. Biotechnol. 76 (2001) 1154-1158.
[48] Y.-Q. Xie, Y.-B. Huang, J.-S. Liu, L.-Y. Ye, L.-M. Che, S. Tu, C.-L. Liu, Pest
Manag. Sci. 71 (2015) 404-414.
26

Scheme 1. The synthetic routes of the intermediates Ia-b and IIIa-b.
27

Scheme 2. The synthetic routes of the title compounds VIIIa-g ang IXa-p.
28

Scheme 3. The synthetic routes of the title compounds Xa-d.
29