Site-selective reactions of hydrazonoyl chlorides with cyanoacetic hydrazide and its N-arylidene derivatives and anti-aggressive activity of prepared products

Article abstract of DOI:10.1007/s12272-013-0082-x

Reactions of hydrazonoyl halides with cyanoacetic hydrazide and its N-arylidene derivatives proceeded site-selectively and afforded the respective pyrazolo[3,4-d]pyridazine and aldehyde N-(1-aryl-3-acetyl-4-cyanopyrazol-5-yl) hydrazone derivatives. The st

Full text of DOI:10.1007/s12272-013-0082-x

Arch. Pharm. Res. (2013) 36:694–701
DOI 10.1007/s12272-013-0082-x
RESEARCH ARTICLE
Site-selective reactions of hydrazonoyl chlorides with cyanoacetic
hydrazide and its N-arylidene derivatives and anti-aggressive
activity of prepared products
•
Ahmad Sami Shawali Thoraya A. Farghaly
•
•
Shadia M. Hussein Mohamed M. Abdalla
Published online: 13 March 2013
Ó The Pharmaceutical Society of Korea 2013
Abstract Reactions of hydrazonoyl halides with cyano-
acetic hydrazide and its N-arylidene derivatives proceeded
site-selectively and afforded the respective pyrazolo[3,4-
d]pyridazine and aldehyde N-(1-aryl-3-acetyl-4-cyanopyr-
azol-5-yl)hydrazone derivatives. The structures of the
products were elucidated on the basis of their spectral and
elemental analyses. The anti-aggressive activity of the
compounds prepared was screened.
these findings and in conjunction with our on going studies
of the chemistry of hydrazonoyl halides (Shawali et al.
1980, 1983, 1993, 1995, 2001, 2003, 2006, 2007, 2008,
2009, 2010a, b, c), it was thought interesting to study the
reactions of the hydrazonoyl chlorides 1 with each of cy-
anoacetic hydrazide 6 and aldehyde N-cyanoacetylhyd-
razones 7. Our objective after such a study is to shed some
light on the site selectivity in the target reactions. This is
because the latter reagents each has more than one reactive
sites namely the active methylene (NC–CH2–) moiety and
the dipolarophilic C=N group as well as the nucleophilic
hydrazino (–NHNH2) group and thus its reaction with the
halides 1 can theoretically lead to the formation of more
than one products. In addition, in view of the various
reported biological activities exhibited by N-pyra-
zolylhydrazones (Zheng et al. 2009; Xia et al. 2008) and
hydrazones (Liu et al. 2009; Horiuchi et al. 2009; Gue-
tzoyan et al. 2010; Rollas and Kucukguzel 2007), the
bioactivity of the products of the target reactions were
studied.
Keywords Site-selectivity Á Hydrazonoyl halides Á
Pyrazoles Á Pyrazolo[3,4-d] pyridazines Á
Anti-aggressive activity
Introduction
Hydrazonoyl halides 1 were reported to react with active
methylene nitriles such as cyanoacetophenone, alkyl cya-
noacetates, cyanoacetanilides and malononitrile in etha-
nolic sodium ethoxide to form the respective pyrazole
derivatives 2–4 (Scheme 1) (Shawali and Parkanyi 1980).
Also, it was reported that reaction of hydrazonoyl halides 1
with several acyclic and cyclic ketone N-substituted
hydrazones gave the corresponding 4,5-dihydro-1,2,4-
triazoles 5 via cycloaddition of the nitrilimine intermediate
to the C=N of the hydrazone residue (Scheme 1) (Awad
et al. 2002; Awadallah et al. 2002,2004). In the light of
Materials and methods
All melting points were determined on an electrothermal
Gallenkamp apparatus. Solvents were generally distilled
and dried by standard literature procedures prior to use.
The IR spectra were measured on a Pye-Unicam SP300
1
instrument in potassium bromide discs. The H and 13C
NMR spectra were recorded in DMSO-d6 on a Varian
Mercury VXR spectrometer (300 MHz for 1H and 75 MHz
for 13C NMR) and the chemical shifts d downfield from
tetramethylsilane (TMS) as an internal standard. The mass
spectra were recorded on a GCMS-Q1000-EX Shimadzu
and GCMS 5988-A HP spectrometers, the ionizing voltage
A. S. Shawali (&) Á T. A. Farghaly Á S. M. Hussein
Department of Chemistry, Faculty of Science,
University of Cairo, Giza 12613, Egypt
e-mail: as_shawali@mail.com
M. M. Abdalla
Research Unit, Saco Pharm. Co, 6th October City, Giza, Egypt
123

Site-selective reactions of hydrazonoyl chlorides with cyanoacetic hydrazide
695
Scheme 1 Reaction of active
methylene nitriles and ketone N-
substituted hydrazones with
Ar
H
N
X
N
ii
i
R
hydrazonoyl halides
Et3N
EtONa / EtOH
1
R
R
N
z
X = Cl. Br
NHR"'
N
N
R'
N
N
Y
R"
Ar
5
Ar
2- 4
i = PhCOCH2CN; NCCH2COOEt; NCCH2CONHPh or CH2(CN)2
R'/R" : alkyl or cycloalkyl
ii = R'R"C=NNHR"'
R"' = MeCO, EtOCO, PhCO
Z / Y : 2, NC / Ph; 3a, COOEt / NH2; 3b, PhNHCO / NH2; 4, NC / NH2
R = MeCO, PhCO, 2-Naphthoyl, MeOCO
3-Amino-7-methyl-2-(4-methylphenyl)-2,5-dihydro-4-
was 70 eV. Elemental analyses were carried out by the
Microanalytical Center of Cairo University, Giza, Egypt.
Hydrazonoyl halides 1 (Shawali and Parkanyi 1980),
2-Cyanoacetic hydrazide 6 and N-arylidene cyanoaceti-
chydrazide 7a–e (Geissler et al. 1982; Duffin and Kendall
1960; Al-Awadi et al. 1996) were prepared as previously
described.
oxo-pyrazolo[3,4-d]pyridazine (10b)
Yellow solid, (92 % yield), mp: 240–242 °C (EtOH); IR
(KBr) mmax/cm-1 3317 (NH), 3186, 3116 (NH2), 1689
1
(C=O); H NMR: d 2.17 (s, 3H, CH3), 2.24 (s, 3H, CH3),
4.15 (s, 2H, NH2), 7.07 (d, J = 9 Hz, 2H, Ar–H), 7.21 (d,
J = 9 Hz, 2H, Ar–H), 10.05 (s, 1H, NH); 13C NMR: d
14.26, 25.21, 115.83, 116.45, 124.54, 125.16, 129.49,
142.97, 144.99, 156.82, 166.38; MS m/z (%): 255 (M?,
67), 254 (21), 133 (29), 124 (45), 106 (100), 91 (38), 78
(71), 77 (48). Anal. Calcd. for C13H13N5O (255.28): C,
61.17; H, 5.13; N, 27.43. Found: C, 61.05; H, 5.09; N,
27.26 %.
Reaction of hydrazonoyl halides with cyanoacetic
hydrazide
General procedure
To a stirred solution of 2-cyanoacetic hydrazide 6 (0.5 g,
5 mmol) and the appropriate hydrazonoyl chloride 1
(5 mmol) in dry dioxane (30 ml), triethylamine (0.7 ml)
was added and the mixture was refluxed for 5 h. The solid
product, so formed in each case, was collected by filtration,
washed with water, dried, and crystallized from ethanol to
afford the corresponding pyrazolo[3,4-d]pyridazine deriv-
atives 10. The compounds 10a–d prepared are listed below
together with their physical constants.
3-Amino-7-methyl-2-(4-chlorophenyl)-2,5-dihydro-4-
oxo-pyrazolo[3,4-d]pyridazine (10c)
Dark orange solid, (89 % yield), mp: 148–50 °C (EtOH);
IR (KBr) mmax/cm-1 3259 (NH), 3186, 3105 (NH2), 1685
1
(C=O); H NMR: d 2.15 (s, 3H, CH3), 4.02 (s, 2H, NH2),
7.21 (d, J = 9 Hz, 2H, Ar–H), 7.33 (d, J = 9 Hz, 2H, Ar–
H), 10.21 (s, 1H, NH); MS m/z (%): 277 (M??2, 16), 275
(M?, 40), 212 (25), 149 (66), 125 (59), 111 (45), 105 (44),
83 (50), 76 (19). Anal. Calcd. for C12H10ClN5O (275.70):
C, 52.28; H, 3.66; N, 25.40. Found: C, 52.09; H, 3.54; N,
25.19 %.
3-Amino-7-methyl-2-phenyl-2,5-dihydro-4-oxo-
pyrazolo[3,4-d] pyridazine (10a)
Dark orange solid, (90 % yield), mp: 242–244 °C (EtOH);
IR (KBr) mmax/cm-1 3317 (NH), 3193, 3109 (NH2), 1693
1
3-Amino-7-methyl-2-(4-nitrophenyl)-2,5-dihydro-4-
(C=O); H NMR: d 2.22 (s, 3H, CH3), 4.32 (s, 2H, NH2),
oxo-pyrazolo[3,4-d]pyridazine (10d)
6.69-7.37 (m, 5H Ar–H), 10.08 (s, 1H, NH); MS m/z (%):
242 (M??1, 26), 241 (M?, 24), 174 (21), 146 (18), 123
(24), 106 (26), 91 (100), 77 (42). Anal. Calcd. for
C12H11N5O (241.25): C, 59.74; H, 4.60; N, 29.03. Found:
C, 59.61; H, 4.48; N, 29.14 %.
Yallow solid, (85 % yield), mp: 286–288 °C (EtOH); IR
(KBr) mmax/cm-1 3448 (NH), 3232, 3124 (NH2), 1685
1
(C=O); H NMR: d 2.24 (s, 3H, CH3), 4.14 (s, 2H, NH2),
123

696
A. S. Shawali et al.
4-Chlorobenzaldehyde N-[1-(4-chlorophenyl)-3-acetyl-
4-cyanopyrazol-5-yl]-hydrazone (15c)
7.45 (d, J = 8 Hz, 2H, Ar–H), 8.16 (d, J = 8 Hz, 2H, Ar–
H), 10.88 (s, 1H, NH); MS m/z (%): 287 (M??1, 34), 286
(M?, 77), 250 (34), 222 (23), 208 (31), 180 (34), 149 (57),
124 (69), 106 (43, 91 (40), 83 (43), 77 (46). Anal. Calcd.
for C12H10N6O3 (286.25): C, 50.35; H, 3.52; N, 29.36.
Found: C, 50.21; H, 3.39; N, 29.20 %.
Orange solid, (80 % yield), mp: 262–264 °C (EtOH); IR
(KBr) mmax/cm-1 3205 (NH), 2221 (CN), 1709 (C=O); H
1
NMR: d 2.35 (s, 3H, COCH3), 7.21 (d, J = 9 Hz, 2H,
ArH), 7.37 (d, J = 9 Hz, 2H, Ar–H), 7.54 (d, J = 9 Hz,
2H, ArH), 7.85 (d, J = 9 Hz, 2H, Ar–H), 9.60 (s, 1H,
=CH), 11.50 (s, 1H, NH); MS m/z (%): 402 (M??4, 1.2),
401 (M??3, 5), 400 (M??2, 5), 398 (M?, 17), 397 (32),
396 (25), 259 (29), 139 (54), 113 (33), 111 (100), 91 (13),
89 (17), 77 (14). Anal. Calcd. for C19H13Cl2N5O (398.25):
C, 57.30; H, 3.29; N, 17.59. Found: C, 57.25; H, 3.12; N,
17.47 %.
Reaction of hydrazonoyl chlorides 1 with N-arylidene
cyanoacetichydrazides 7
General method
To sodium ethoxide solution, prepared from sodium metal
(0.12 g, 5 mol) and absolute ethanol (30 mL), was added
the appropriate compound 7 (5 mol). The mixture was
stirred for 10 min. To the resulting solution was added the
appropriate hydrazonoyl chloride 1 (5 mol of each) and the
reaction mixture was left overnight at room temperature,
while being stirred. The solid, that precipitated, was filtered
off, washed with water, dried and finally crystallized from
the appropriate solvent to give the respective 15. The
compounds 15a–p prepared together with their physical
constants are listed below.
4-Chlorobenzaldehyde N-[1-(4-nitrophenyl)-3-acetyl-
4-cyanopyrazol-5-yl]hydrazone (15d)
Brown solid, (78 % yield), mp: 200–202 °C (EtOH); IR
(KBr) mmax/cm-1 3220 (NH), 2229 (CN), 1712 (C=O); H
1
NMR: d 2.38 (s, 3H, COCH3), 7.22 (d, J = 9 Hz, 2H,
ArH), 7.42 (d, J = 9 Hz, 2H, Ar–H), 7.58 (d, J = 9 Hz,
2H, ArH), 8.23 (d, J = 9 Hz, 2H, Ar–H), 9.50 (s, 1H,
=CH), 11.70 (s, 1H, NH); MS m/z (%): 410 (M??2, 8),
408 (M?, 14), 391 (14), 154 (20), 137 (100), 111 (8), 110
(14), 108 (26), 102 (41), 92 (30), 89 (17), 77 (21). Anal.
Calcd. for C19H13ClN6O3 (408.81): C, 55.82; H, 3.21; N,
20.56. Found: C, 55.79; H, 3.10; N, 20.38 %.
4-Chlorobenzaldehyde N-[1-phenyl-3-acetyl-4-
cyanopyrazol-5-yl]-hydrazone (15a)
Orange solid, (89 % yield), mp: 230–232 °C (EtOH/
Dioxane); IR (KBr) mmax/cm-1 3232 (NH), 2218 (CN),
1701 (C=O); H NMR: d 2.34 (s, 3H, COCH3), 6.73–7.48
4-Chlorobenzaldehyde N-[1-(4-acetylphenyl)-3-acetyl-
4-cyanopyrazol-5-yl]-hydrazone (15e)
1
(m, 5H Ar–H), 7.63 (d, J = 9 Hz, 2H, ArH), 7.89 (d,
J = 9 Hz, 2H, Ar–H), 9.61 (s, 1H, =CH), 11.55 (s, 1H,
NH); MS m/z (%): 365 (M??2, 9), 363 (M?, 23), 225 (17),
138 (13), 111 (6), 105 (33), 91 (11), 77 (100). Anal. Calcd.
for C19H14ClN5O (363.81): C, 62.73; H, 3.88; N, 19.25.
Found: C, 62.54; H, 3.59; N, 19.16 %.
Brown solid, (80 % yield), mp: 282–284 °C (EtOH); IR
(KBr) mmax/cm-1 3236 (NH), 2225 (CN), 1697, 1674
(2C=O); H NMR: d 2.37 (s, 3H, COCH3), 2.46(s, 3H,
1
COCH3), 7.25 (d, J = 9 Hz, 2H, ArH), 7.45 (d, J = 9 Hz,
2H, Ar–H), 7.63 (d, J = 9 Hz, 2H, ArH), 7.96 (d,
J = 9 Hz, 2H, Ar–H), 9.56 (s, 1H, =CH), 11.64 (s, 1H,
NH); MS m/z (%): 407 (M??2, 30), 405 (M?, 49), 404
(27), 267 (24), 137 (63), 119 (100), 111 (16), 102 (35), 91
(71), 89 (27), 77 (38). Anal. Calcd. for C21H16ClN5O2
(405.85): C, 62.15; H, 3.97; N, 17.26. Found: C, 62.29; H,
3.88; N, 17.15 %.
4-Chlorobenzaldehyde N-[1-(4-methylphenyl)-3-
acetyl-4-cyanopyrazol-5-yl]-hydrazone (15b)
Orange solid, (87 % yield), mp: 270–272 °C (EtOH); IR
1
(KBr) mmax/cm-1 3217 (NH), 2218 (CN), 1697 (C=O); H
Benzaldehyde N-(1-phenyl-3-acetyl-4-cyanopyrazol-
5-yl)hydrazone (15f)
NMR: d 2.27 (s, 3H, CH3), 2.35 (s, 3H, COCH3), 6.70–7.25
(m, 4H, Ar–H), 7.31 (d, J = 9 Hz, 2H, ArH), 7.41 (d,
J = 9 Hz, 2H, Ar–H), 9.60 (s, 1H, =CH), 11.90 (s, 1H,
NH); MS m/z (%): 379 (M??2, 13), 377 (M?, 21), 239
(14), 139 (33), 138 (60), 136 (100), 119 (29), 111 (27), 102
(21), 91 (82), 89 (46), 77 (16). Anal. Calcd. for
C20H16ClN5O (377.84): C, 63.58; H, 4.27; N, 18.54.
Found: C, 63.62; H, 4.39; N, 18.67 %.
Orange solid, (82 % yield), mp: 200–202 °C (EtOH); IR
(KBr) mmax/cm-1 3209 (NH), 2218 (CN), 1705 (C=O); H
1
NMR: d 2.33 (s, 3H, COCH3), 6.71–7.88 (m, 10H, Ar–H),
9.61 (s, 1H, =CH), 11.65 (s, 1H, NH); MS m/z (%): 330
(M??1, 9), 329 (M?, 30), 328 (17), 225 (16), 105 (31), 77
123

Site-selective reactions of hydrazonoyl chlorides with cyanoacetic hydrazide
697
4-Methoxylbenzaldehyde N-[1-(4-methylphenyl)-3-
acetyl-4-cyano-pyrazol-5-yl]-hydrazone (15k)
(100). Anal. Calcd. for C19H15N5O (329.36): C, 69.29; H,
4.59; N, 21.26. Found: C, 69.15; H, 4.42; N, 21.18 %.
Redish brown solid, (80 % yield), mp: 210–212 °C
(EtOH); IR (KBr) mmax/cm-1 3217 (NH), 2214 (CN), 1689
(C=O); 1H NMR: d 2.27 (s, 3H, CH3), 2.35 (s, 3H,
COCH3), 3.80 (s, 3H, OCH3), 6.87 (d, J = 9 Hz, 2H,
ArH), 6.98 (d, J = 9 Hz, 2H, Ar–H), 7.12 (d, J = 9 Hz,
2H, ArH), 7.62 (d, J = 9 Hz, 2H, Ar–H), 9.45 (s, 1H,
=CH), 11.65 (s, 1H, NH); MS m/z (%): 374 (M??1, 3),
373 (M?, 11), 217 (21), 133 (100), 103 (11), 91 (33), 77
(29). Anal. Calcd. for C21H19N5O2 (373.42): C, 67.55; H,
5.13; N, 18.75. Found: C, 67.39; H, 5.27; N, 18.60 %.
4-Methoxybenzaldehyde N-(1-phenyl-3-acetyl-4-
cyanopyrazol-5-yl)-hydrazone (15g)
Brown solid, (84 % yield), mp: 212–214 °C (EtOH); IR
(KBr) mmax/cm-1 3217 (NH), 2222 (CN), 1697 (C=O); H
1
NMR: d 2.36 (s, 3H, COCH3), 3.80 (s, 3H, OCH3), 6.98 (d,
J = 9 Hz, 2H, ArH), 7.0–7.40 (m, 5H, Ar–H), 7.62 (d,
J = 9 Hz, 2H, Ar–H), 9.50 (s, 1H, =CH), 11.64 (s, 1H,
NH); MS m/z (%): 359 (M?, 17), 358 (42), 151 (50), 69
(75). Anal. Calcd. for C20H17N5O2 (359.39): C, 66.84; H,
4.77; N, 19.49. Found: C, 66.89; H, 4.67; N, 19.36 %.
4-Methylbenzaldehyde N-[1-(4-methylphenyl)-3-
acetyl-4-cyanopyrazol-5-yl]-hydrazone (15l)
4-Nitrobenzaldehyde N-(1-phenyl-3-acetyl-4-
cyanopyrazol-5-yl)-hydrazone (15h)
Orange solid, (82 % yield), mp: 262–264 °C (EtOH); IR
Orange solid, (75 % yield), mp: 252–254 °C (EtOH); IR
1
(KBr) mmax/cm-1 3209 (NH), 2218 (CN), 1697 (C=O); H
1
(KBr) mmax/cm-1 3247 (NH), 2221 (CN), 1705 (C=O); H
NMR: d 2.25 (s, 3H, CH3), 2.29 (s, 3H, CH3), 2.34 (s, 3H,
COCH3), 6.55 (d, J = 8 Hz, 2H, ArH), 6.85 (d, J = 8 Hz,
2H, Ar–H), 7.29 (d, J = 8 Hz, 2H, ArH), 7.37 (d,
J = 9 Hz, 2H, Ar–H), 9.54 (s, 1H, =CH), 11.62 (s, 1H,
NH); MS m/z (%): 357 (M?, 30), 238 (18), 118 (32), 91
(100), 77 (14). Anal. Calcd. for C21H19N5O (357.42): C,
70.57; H, 5.36; N, 19.59. Found: C, 70.42; H, 5.48; N,
19.41 %.
NMR: d 2.37 (s, 3H, COCH3), 6.91–7.42 (m, 5H, Ar–H),
7.49 (d, J = 9 Hz, 2H, ArH), 8.15 (d, J = 9 Hz, 2H, Ar–
H), 8.86 (s, 1H, =CH), 11.50 (s, 1H, NH); MS m/z (%): 375
(M??1, 6), 374 (M?, 30), 373 (15), 225 (16), 119 (12),
105 (33), 92 (15), 77 (100). Anal. Calcd. for C19H14N6O3
(374.36): C, 60.96; H, 3.77; N, 22.45. Found: C, 60.87; H,
3.65; N, 22.29 %.
4-Methylbenzaldehyde N-(1-phenyl-3-acetyl-4-
cyanopyrazol-5-yl)-hydrazone (15i)
4-Methylbenzaldehyde N-[1-(4-nitrophenyl)-3-acetyl-
4-cyanopyrazol-5-yl]-hydrazone (15m)
Yellow solid, (80 % yield), mp: 242–244 °C (EtOH); IR
1
(KBr) mmax/cm-1 3224 (NH), 2225 (CN), 1705 (C=O); H
Brown solid, (79 % yield), mp: 270–272 °C (EtOH); IR
1
(KBr) mmax/cm-1 3201 (NH), 2225 (CN), 1708 (C=O); H
NMR: d 2.24 (s, 3H, CH3), 2.36 (s, 3H, COCH3), 6.60–7.13
(m, 5H, Ar–H), 7.28 (d, J = 9 Hz, 2H, ArH), 7.72 (d,
J = 9 Hz, 2H, Ar–H), 9.54 (s, 1H, =CH), 11.65 (s, 1H,
NH); MS m/z (%): 344 (M??1, 10), 343 (M?, 43), 225
(25), 118 (38), 104 (12), 91 (32), 77 (100). Anal. Calcd. for
C20H17N5O (343.39): C, 69.96; H, 4.99; N, 20.39. Found:
C, 69.74; H, 4.85; N, 20.26 %.
NMR: d 2.25 (s, 3H, CH3), 2.40 (s, 3H, COCH3), 7.09 (d,
J = 9 Hz, 2H, ArH), 7.59 (d, J = 9 Hz, 2H, Ar–H), 7.72
(d, J = 9 Hz, 2H, ArH), 8.18 (d, J = 9 Hz, 2H, Ar–H),
9.46 (s, 1H, =CH), 11.80 (s, 1H, NH); MS m/z (%): 388
(M?, 41), 270 (20), 224 (10), 150 (25), 118 (100), 77 (80).
Anal. Calcd. for C20H16N6O3 (388.39): C, 61.85; H, 4.15;
N, 21.64. Found: C, 61.66; H, 4.01; N, 21.55 %.
Benzaldehyde N-[1-(4-methylphenyl)-3-acetyl-4-
cyanopyrazol-5-yl]-hydrazone (15j)
4-Methylbenzaldehyde N-[1-(4-chlorophenyl)-3-acetyl-
4-cyanopyrazol-5-yl]-hydrazone (15n)
Orange solid, (89 % yield), mp: 240–242 °C (EtOH); IR
1
(KBr) mmax/cm-1 3205 (NH), 2218 (CN), 1693 (C=O); H
Orange solid, (85 % yield), mp: 250–252 °C (EtOH); IR
1
(KBr) mmax/cm-1 3190 (NH), 2218 (CN), 1705 (C=O); H
NMR: d 2.28 (s, 3H, CH3), 2.35 (s, 3H, COCH3), 6.67–7.42
(m, 5H, Ar–H), 7.62 (d, J = 9 Hz, 2H, ArH), 7.86 (d,
J = 9 Hz, 2H, Ar–H), 9.60 (s, 1H, =CH), 11.52 (s, 1H,
NH); MS m/z (%): 343 (M?, 27), 342 (22), 238 (17), 105
(15), 91 (100), 77 (28). Anal. Calcd. for C20H17N5O
(343.39): C, 69.96; H, 4.99; N, 20.39. Found: C, 70.12; H,
4.84; N, 20.30 %.
NMR: d 2.25 (s, 3H, CH3), 2.31 (s, 3H, COCH3), 7.11 (d,
J = 8 Hz, 2H, ArH), 7.16 (d, J = 8 Hz, 2H, Ar–H), 7.36
(d, J = 8 Hz, 2H, ArH), 7.43 (d, J = 8 Hz, 2H, Ar–H),
9.51 (s, 1H, =CH), 11.59 (s, 1H, NH); MS m/z (%): 379
(M??2, 15), 377 (M?, 61), 376 (58), 259 (39), 139 (44),
119 (51), 118 (100), 111 (82), 110 (76), 91 (55), 77 (15).
123

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A. S. Shawali et al.
Anal. Calcd. for C20H16ClN5O (377.84): C, 63.58; H, 4.27;
N, 18.54. Found: C, 63.38; H, 4.08; N, 18.36 %.
fights is recorded for each pair during the 3-min period.
The fighting behavior consists of vocalization, leaping,
running, rearing and facing each other with some attempt
to attack by hitting, biting or boxing. The test compound or
the standard are applied either 30 min before the test i.p. or
60 min before the test orally. For a time response, the drug
is given 30, 60 and 120 min prior to testing. Six pairs of
drug-treated and two pairs of vehicle-treated animals are
utilized for each time period. A dose range is tested at the
peak of drug activity. A minimum of three doses (10 pairs
of mice/dose) is administered for a range of doses. Control
animals receive the vehicle.
4-Methoxylbenzaldehyde N-[1-(4-chlorophenyl)-3-
acetyl-4-cyano-pyrazol-5-yl]-hydrazone (15o)
Orange solid, (80 % yield), mp: 210–212 °C (EtOH); IR
1
(KBr) mmax/cm-1 3213 (NH), 2221 (CN), 1705 (C=O); H
NMR: d 2.35 (s, 3H, COCH3), 3.80 (s, 3H, OCH3), 6.97 (d,
J = 9 Hz, 2H, ArH), 7.10 (d, J = 9 Hz, 2H, Ar–H), 7.42
(d, J = 9 Hz, 2H, ArH), 7.62 (d, J = 9 Hz, 2H, Ar–H),
7.94 (s, 1H, =CH), 11.63 (s, 1H, NH); MS m/z (%): 395
(M??2, 9), 393 (M?, 34), 392 (21), 260 (11), 259 (12),
135 (100), 111 (28), 91 (17), 77 (25). Anal. Calcd. For
C20H16ClN5O2 (393.84): C, 61.00; H, 4.09; N, 17.78.
Found: C, 60.95; H, 4.21; N, 17.56 %.
Results and discussion
Reactions of cyanoacetic hydrazide 6 with the hydrazonoyl
chlorides 1a–d were examined. When such reactions were
carried out in refluxing dioxane in the presence of trieth-
ylamine, they yielded, in each case, one product whose
4-Nitrobenzaldehyde N-[1-(4-methylphenyl)-3-acetyl-
4-cyanopyrazol-5-yl]-hydrazone (15p)
1
Red solid, (84 % yield), mp: 280–282 °C (EtOH); IR
spectra (IR, H NMR and mass) and elemental analysis
(KBr) mmax/cm-1 3240 (NH), 2218 (CN), 1697 (C=O); H
data are consistent with structure 10 rather than 12 and 14
(Scheme 2). For example, their IR spectra were free of any
bands due to nitrile absorption. They showed instead three
characteristic absorption bands in the regions v 3448–3317,
3232–3105 and 1693–1685 cm-1 assignable to the CONH,
NH2 and CONH groups, respectively. The 1H NMR spectra
showed, in addition to the aromatic proton signals, three
characteristic signals in the regions d 2.24–2.15, 4.32–4.02
and 10.88–10.05 assignable to the CH3, NH2, and –CONH–
protons, respectively. The 13C NMR spectra of 10b taken
as typical example of the series prepared, revealed the
signal of the carbonyl carbon of the pyrimidinone ring
residue at d 166.38 ppm rather than the carbonyl carbon of
acetyl group of compound 12. On the basis of these data,
the other possible structures 12 and 14 were discarded. To
account for the formation of 10, it is suggested, as depicted
in Scheme 2, that the reaction starts with the formation of
the substitution intermediate 8 which in turn undergoes two
tandem in situ cyclization to give the respective pyrazol-
o[3,4-d]pyridazine derivatives 10 as end products.
1
NMR: d 2.27 (s, 3H, CH3), 2.35 (s, 3H, COCH3), 7.21 (d,
J = 8 Hz, 2H, ArH), 7.32 (d, J = 8 Hz, 2H, Ar–H), 8.14
(d, J = 9 Hz, 2H, ArH), 8.35 (d, J = 9 Hz, 2H, Ar–H),
8.86 (s, 1H, =CH), 12.05 (s, 1H, NH); MS m/z (%): 389
(M??1, 9), 388 (M?, 28), 239 (15), 119 (23), 106 (13), 91
(100), 77 (15). Anal. Calcd. for C20H16N6O3 (388.39): C,
61.85; H, 4.15; N, 21.64. Found: C, 61.68; H, 3.99; N,
21.42 %.
Measurement of anti-aggressive activity
Since the discovery of the taming effects of benzodiaze-
pines in vicious monkeys (Heise and Boff 1961; Randall
et al. 1961) tests for agents with anti-aggressive activity
have been developed for various animal species. These
tests include foot-shock induced aggression in mice and
rats, fighting behavior of isolated mice and aggressiveness
of rats which become extremely vicious after lesions in the
septal area of the brain (Brady and Nauta 1953). Foot-
shock induced aggression is used for further characteriza-
tion of centrally active drugs. Irwin et al. (1971) have
attempted to compare drug classes with this method.
Male mice (NMRI, Ivanovas) with a weight between 20
and 30 g are used. Two mice are placed in a box with a grid
floor consisting of steel rods with a distance of 6 mm. A
constant current of 0.6 or 0.8 mA is supplied to the grid
floor by a LVE constant current shocker with an associated
scrambler. A 60-Hz current is delivered for 5 s followed by
5 s intermission for 3 min. Each pair of mice is dosed and
tested without previous exposure. The total number of
Next, reactions of hydrazonoyl halides 1a–f with N-a-
rylidenecyanoacetic hydrazides 7a–e were examined.
When each of the hydrazones 7a–e was treated with the
appropriate hydrazonoyl chloride 1 in ethanol in the pres-
ence of sodium ethoxide at room temperature, it yielded, in
each case, a single product identified as the respective
aldehyde N-(1-aryl-3-acetyl-4-cyanopyrazol-5-yl)hydra-
zone 15 (Scheme 3). The structures of the products 15 were
elucidated on the basis of their spectral and elemental
analysis data. For example, their IR spectra showed in each
case three characteristic bands in the regions 3217–3247,
2214–2229 and 1697–1712 cm-1 assignable to the
123

Site-selective reactions of hydrazonoyl chlorides with cyanoacetic hydrazide
699
Scheme 2 Reaction of
cyanoacetic hydrazide 6 with
the hydrazonoyl chlorides 1a–d
Cl
NC-CH2-CO-NHNH2
+
CH3CO
NNHAr
- HCl
6
1
Et3N / Dioxane / heat
H
CH3CO
CN
NC-CH2-CO-NH-NH
CONHNH2
N
ArNHN
13
COCH3
NH
Ar
8
- HOH
O
H
CH3CO
CN
N
COCH3
N
CN
HN
NC
N
N
NHNH2
N
N
N
NNH2
9
Ar
Ar
CH3
O
11
NH2
14
CH3CO
NH2
Ar = 4-XC6H4;
X : a, H; b, Me; c, Cl; d, NO2
N
N
HN
N
N
Ar
N
N
N
H
Ar
12
CH3
10
hydrazone NH, cyano and acetyl carbonyl groups, respec-
1
tively. Also, their H NMR spectra showed, in each case,
showed potent activities. For compounds 7, the order of
activity is 7c [ 7d [ 7e [ 7b. This finding indicates that
electron withdrawing groups such as Cl, NO2 and COCH3
increase the activity, whereas electron donating substitu-
ents such as methyl group decrease the reactivity.
For compounds 15, the order of activity is 15 h [ 15a
[ 15i [ 15o [ 15c [ 15n [ 15p [ 15m [ 15k [ 15e [
15j [ 15g [ 15f [ 15b. This result indicates also that the
presence of unsubstituted phenyl group at position-1 of the
pyrazole ring residue and electron withdrawing substituent
such as NO2 and Cl at the phenylhydrazone moiety increase
the activity.
two singlet signals in the regions d 9.45–9.61 and
11.50–12.05 assignable to the ArCH=N and –NH–N=
protons, respectively. To account for the formation of the
products 15, it is suggested, as depicted in Scheme 3, that
reaction of 1 with 7 starts with nucleophilic substitution to
give B as intermediate which underwent in situ cyclization
via elimination of water to give 15 as end products. This
sequence is compatible with literature reports on reactions
of hydrazonoyl halides with cyanoacetophenone (Shawali
and Parkanyi 1980). Based on the foregoing results, it is
clear that the studied reactions are site selective.
From the above results, it can be concluded that the
hydrazone derivatives give high activity similar to the open
straight chain of most serotonin re-uptake inhibitors. Also,
the electron cloud on the phenyl moiety seems to play an
important role in increasing the activity of compounds 7
and 15 and that the electron withdrawing substituents
increase the activity more than the electron donating
groups.
Biological results
The inhibition percent of aggression is calculated from the
vehicle control and ED50 values are calculated (Table 1).
As shown in the latter table, all of the tested compounds
123

700
A. S. Shawali et al.
Scheme 3 Reaction of
hydrazonoyl chlorides 1 with N-
arylidenecyanoacetic
hydrazides 7
O
Cl
1
H
N
NC
N
7
N
H
EtONa / EtOH
MeCO
N
+
X
Y
MeCO
CN
H
MeCO
CN
O
H
N
N
N
H
N
Y
NH
N
N
N
- H2O
Y
X
15
B
X
1, Y: a, H; b, Me; c, OMe; d, Cl; e, NO2; f, COMe
7, X: a, H; b, Me; c, Cl; d, NO2; e, OMe
15
X
15
15
Y
Y
X
X
Y
H
Me
Cl
NO2
COMe
H
g
h
i
j
k
l
a
b
c
d
e
f
Cl
OMe
NO2
Me
H
OMe
Me
H
H
H
Me
Me
Me
m
NO2
Cl
Cl
Me
Me
OMe
NO2
Cl
Cl
Cl
Cl
H
n
o
p
Me
Table 1 Anti-aggressive
activity of compounds 7
and 15a–p
Compound no.
The percent inhibition
of aggression (%)
ED50 (lg/kg)a
LD50 (lg/kg)b
7b
79.38
98.12
97.56
91.52
96.87
81.28
95.55
77.12
92.48
85.68
88.18
98.79
96.58
90.13
93.48
94.48
95.12
95.89
94.89
0.532 0.0022 910-3
0.117 0.001 910-3
0.118 0.0011 910-3
0.201 0.0012 910-3
0.122 0.0023 910-3
0.512 0.0012 910-3
0.139 0.0012 910-3
0.562 0.0034 910-3
0.198 0.0022 910-3
0.414 0.0013 910-3
0.392 0.0013 910-3
0.110 0.001 910-3
0.129 0.0034 910-3
0.211 0.0012 910-3
0.177 0.0034 910-3
0.161 0.0056 910-3
0.142 0.0013 910-3
0.131 0.0012 910-3
0.155 0.0012 910-3
475.46 4.44
765.78 6.55
678.432 7.54
384.00 9.32
678.55 7.45
398.54 4.66
432.54 5.54
267.54 5.33
345.99 9.54
456.43 5.44
395.34 6.33
567.556 3/45
345.67 9.66
375.00 7.22
345.68 8.78
443.55 8.66
453.56 6.43
234.89 2.78
456.43 7.45
7c
7d
7e
15a
15b
15c
15d
15e
15f
15g
15h
15i
15j
15k
15m
15n
15o
15p
a
ED50 is the dose that produces
50 % of the maximal response
to the compound
b
LD50 is the median lethal dose
and it is the amount of the
compound required (usually per
body weight) to kill 50 % of the
test population
123

Site-selective reactions of hydrazonoyl chlorides with cyanoacetic hydrazide
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Conclusion
Reactions of hydrazonoyl chlorides with cyanoacetic
hydrazide and its N-arylidene derivatives proceeded site-
selectively and afforded the respective pyrazolo[3,4-
d]pyridazine and aldehyde N-(1-aryl-3-acetyl-4-cyanopyr-
azol-5-yl)hydrazone derivatives. The newly synthesized
compounds showed promising anti-aggressive activities.
Rollas, S., and S.G. Kucukguzel. 2007. Biological activities of
hydrazone derivatives. Molecules 12: 1910–1939.
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277–338.
Shawali, A.S., and M.M. Edrees. 2006. Reactions of nitrilimines with
heterocyclic amines and enamines. Convenient methodology for
synthesis and annulation of heterocycles. Arkivoc ix: 292–365.
Shawali, A.S., and S.M. Elsheikh. 2001. Annelated[1,2,4,5]tetrazines.
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