Synthesis of modified steroids as a novel class of non-ulcerogenic, anti-inflammatory and anti-nociceptive agents

Article abstract of DOI:10.1016/j.steroids.2011.05.011

The identification of compounds able to treat both pain and inflammation with limited side effects is one of the prominent goals in biomedical research. This study aimed at the synthesis of new modified steroids with structures justifying non-ulcerogenic,

Full text of DOI:10.1016/j.steroids.2011.05.011

Steroids 76 (2011) 1190–1203
Contents lists available at ScienceDirect
Steroids
journal homepage: www.elsevier.com/locate/steroids
Synthesis of modified steroids as a novel class of non-ulcerogenic,
anti-inflammatory and anti-nociceptive agents
Rafat M. Mohareba,b, Gamal A. Elmegeedc,∗, Omar M.E. Abdel-Salamd, Senot H. Dossc, Marian G. Williamc,1
a Organic Chemistry Department, Faculty of Pharmacy, October University of Modern Sciences and Arts (MSA), Elwahaat Road, October City, Egypt
b Chemistry Department, Faculty of Science, Cairo University, Cario, Egypt
c Hormones Department, National Research Centre, 12622 Dokki, Cairo, Egypt
d Toxicology and Narcotics Department, National Research Centre, 12622 Dokki, Cairo, Egypt
a r t i c l e i n f o
a b s t r a c t
Article history:
The identification of compounds able to treat both pain and inflammation with limited side effects is
one of the prominent goals in biomedical research. This study aimed at the synthesis of new modified
steroids with structures justifying non-ulcerogenic, anti-inflammatory and anti-nociceptive activities.
The steroid derivatives were synthesized via straightforward and efficient methods and their structures
were established based on the analytical and spectral data. The in vivo anti-inflammatory, anti-nociceptive
and anti-ulcerogenic activities of some of these compounds were studied. The newly synthesized com-
pounds 8b, 19b, 24 and 31a showed anti-inflammatory, anti-nociceptive and anti-ulcerogenic activity
with various intensities. Oedema was significantly reduced by either dose 25 or 50 mg/kg of all tested
compounds at 3 and 4 h post-carrageenan. Compound 19b was the most effective in alleviating thermal
pain. The analgesic activity of either dose of the compounds 8b, 24, 31a as well as the high dose 19b was
significantly higher than that for indomethacin (IND). Gastric mucosal lesions caused in the rats by the
administration of 96% EtOH and IND were inhibited by all tested compounds administered at (50 mg/kg)
dose in the study.
Received 17 March 2011
Received in revised form 17 May 2011
Accepted 20 May 2011
Available online 31 May 2011
Keywords:
Anti-inflammatory
Anti-nociceptive
Anti-ulcerogenic
Azoles
Epoxids
Oxarine
© 2011 Elsevier Inc. All rights reserved.
1. Introduction
to an enhanced anti-inflammatory effect become an urgent need
for inflammation patients.
Inflammatory diseases are widely prevalent throughout the
world. Drugs used for the treatment of acute and chronic inflamma-
tory disorders suppress biological processes that contribute to the
onset and symptoms of inflammation [1,2]. Pain is widely accepted
to be one of the most important determinations of quality of life.
A study reported by the World Health Organization demonstrated
that individuals who live with persistent pain suffer fourfold more
from depression or anxiety compared to healthy subjects [3,4]. The
identification of compounds able to treat both pain and inflam-
mation with limited side effects is one of the prominent goals in
biomedical research.
The chronic use of the anti-inflammatory drugs is limited for
their severe side effects such as gastrointestinal injury, espe-
cially gastrointestinal perforation, peptic ulceration or significant
bleeding [5,6]. Thus, developing new therapeutic agents that can
overcome gastrointestinal injury and at the same time could lead
Although the beneficial effects of corticosteroids in the man-
agement of inflammatory and allergic conditions have been
appreciated for over 50 years, complications arising from the
steroid therapy have imposed limitations on the clinical use of this
class of drugs [7]. A considerable research effort has been devoted
to the structural modifications of glucocorticoids, with the hope
of increasing their potencies while minimizing their propensity to
elicit systemic adverse effects. The incorporation of phenylpyra-
zole ring at C-2 and C-3 or fused oxazole ring at C-16 and C-17 [8,9]
increased lipophilicity of glucocorticoids and proved to be useful
for topical application. There has been considerable interest in the
synthesis and biological study of several heterocyclic steroids as
high potent anti-inflammatory agents [10–12].
In recent years our research has focused on the design, synthesis
and pharmacological evaluation of new molecules with analgesic,
anti-inflammatory and antinociceptive activity as potential safe
and effective drugs [12–14]. We report herein on the synthesis
of new steroidal heterocyclic derivatives with structures justi-
fying anti-inflammatory, anti-nociceptive and/or non-ulcerogenic
activities. The in vivo anti-inflammatory, anti-nociceptive and anti-
ulcerogenic activities of some of these compounds were studied.
∗
Corresponding author. Tel.: +20 2 35682070; fax: +20 2 33370931.
E-mail address: gamalae@hotmail.com (G.A. Elmegeed).
Blessing and mercy to the soul of our dear fellow (late) Marian G. William.
1
0039-128X/$ – see front matter © 2011 Elsevier Inc. All rights reserved.
doi:10.1016/j.steroids.2011.05.011

R.M. Mohareb et al. / Steroids 76 (2011) 1190–1203
1191
2. Experimental
100), 256 (24), Calc. for C24H34O3 (370.525): C, 77.80; H, 9.25;
found: C, 78.03; 9.42%.
2.1. Synthetic methods, analytical and spectral data
2.1.1.4. 17-(3ꢀ-Benzoyl-2ꢀ-methyloxiran-2ꢀ-yl)androst-4-ene-3-
one (24). Orange crystals from dioxane, yield 0.32 g (74%), mp
173–175 ◦C, IR (KBr, cm−1): ꢀ = 3030 (CH-aromatic), 2963–2870
(CH-aliphatic), 1710 (C-3, C O), 1723 (COPh), 1649 (C C). 1H NMR
(DMSO-d6, ppm): ı = 0.93 (s, 3H, CH3-19), 1.07 (s, 3H, CH3-18),
1.30 (s, 3H, CH3), 3.87 (s, 1H, oxarine-CH), 5.82 (s, 1H, C4-H),
7.30–7.82 (m, 5H, C6H5), 13C NMR (DMSO-d6, ppm): ı = 35.7 (C-1),
34.0, (C-2), 197.2 (C-3), 125.3 (C-4), 170.9 (C-5), 32.7 (C-6), 31.2
(C-7), 35.4 (C-8), 50.7 (C-9), 37.8 (C-10), 22.0 (C-11), 36.5 (C-12),
39.2 (C-13), 56.3 (C-14), 27.1 (C-15), 22.5 (C-16), 55.8 (C-17), 22.1
(C-18), 20.7 (C-19), 195.6 (C O), 19.4 (CH3), 72.4, 63.0 (C-oxarine),
136.2, 128.7, 129.0, 133.4 (C-phenyl). M.S. (EI): m/z (%): 432 (M+,
43), 327 (M+−COPh, 76), 271 (C19H27O, 54), 161 (34), 105 (100).
Calc. for C29H36O3 (432.594): C, 80.52; H, 8.39; found: C, 80.29; H,
8.66%.
The starting steroid, progesterone, was purchased from Sigma
Company, USA. All solvents were dried by distillation prior to using.
All melting points were measured using an Electrothermal appara-
tus and are uncorrected. The IR spectra were recorded in (KBr discs)
on a shimadzu FT-IR 8201 PC spectrometer and expressed in cm−1
.
The 1H NMR and 13C NMR spectra were recorded with Jeol instru-
ment (Japan), at 270 and 125 MHz, respectively, in DMSO-d6 or
CDCl3 as solvent and chemical shifts were recorded in ppm relative
to TMS. The spin multiplicities were abbreviated by the letters: s-
singlet, d-doublet, t-triplet, q-quartet and m (multiplet, more than
quartet). Mass spectra were recorded on a GCMS-QP 1000 Ex spec-
tra mass spectrometer operating at 70 eV. Elemental analyses were
carried by the Microanalytical Data Unit at the National Research
Center, Giza, Egypt and the Microanalytical Data Unit at Cairo Uni-
versity, Giza, Egypt. The reactions were monitored by thin layer
chromatography (TLC) which was carried out using Merck 60 F254
aluminum sheets and visualized by UV light (254 nm). The mixtures
were separated by preparative TLC and gravity chromatography.
For the nomenclature of steroid derivatives, we used the definitive
rules for the nomenclature of steroids published by the Joint Com-
mission on the Biochemical Nomenclature (JCBN) of IUPAC [15,16].
All described compounds showed the characteristic spectral data
of cyclopentanoperhydrophenanthrene nuclei of pregnene series
and were similar to those reported in the literatures [17,18].
2.1.2. Synthesis of 17-(3ꢀ-arylaminobutanyl)androst-4-ene-3-one
derivatives (16a–c), (16a–c), and (25a–c)
2.1.2.1. General procedure. To a solution of either compound 3
(0.80 g, 2 mmol), compound 15 (0.74 g, 2 mmol), or compound 24
(0.86 g, 2 mmol) in absolute ethanol (30 mL) containing a catalytic
amount of triethylamine (1 mL), either aniline 4a (0.18 g, 2 mmol),
p-chloroaniline 4b (0.25 g, 2 mmol), or p-methoxyaniline 4c (0.24 g,
2 mmol) was added. The reaction mixture, in each case, was heated
under reflux for 6–8 h until all the reactants had disappeared as
indicated by TLC. The reaction mixture poured over an ice/water
mixture and neutralized with dilute hydrochloric acid. The solid
product that formed, in each case, was filtered off, dried and crys-
tallized from the appropriate solvent.
2.1.1. Synthesis of 17-(oxiran-2ꢀ-yl)androst-4-ene-3-one
derivatives (3, 15, and 24)
2.1.1.1. General procedure. To a solution of progesterone 1 (0.31 g,
1 mmol) in potassium tert-butoxide {prepared by the reaction
of anhydrous tert-butanol (30 mL) with potassium metal (0.5 g)
[19]}, equimolar amount of ethyl chloroacetate 2 (0.12 g, 1 mmol),
chloroacetone 14 (0.09 g, 1 mmol), or ␣-bromoacetophenone 23
(0.20 g, 1 mmol) was added. The reaction mixture was stirred at
room temperature for 24 h and then heated for 6 h in water bath
at 70 ◦C. After cooling at room temperature, the reaction mixture
poured over an ice/water mixture and the resulted semisolid was
subjected to extraction with chloroform (2 × 30 mL). The organic
layer was dried over magnesium sulfate and then filtered. The oil
product that formed in each case on removal of the solvent in
vacuum was solidified by boiling in petroleum ether (60–80 ◦C),
collected and crystallized from the appropriate solvent.
2.1.2.2. 17-[Ethyl 3ꢀ-hydroxy-2ꢀ-(phenylamino)butanoate]androst-
4-ene-3-one (5a). Reddish brown powder from methanol, yield
0.67 g (68%), mp 78–80 ◦C, IR (KBr, cm−1): ꢀ = 3546–3387 (OH, NH),
3035 (CH-aromatic), 2970–2878 (CH-aliphatic), 1708 (C-3, C O),
1745 (CO-ester), 1640 (C C). 1H NMR (CDCl3, ppm): ı = 0.96 (s, 3H,
CH3-19), 1.13 (s, 3H, CH3-18), 1.30 (s, 3H, CH3), 1.37 (t, J = 6 Hz, 3H,
ester-CH3), 3.52 (s, 1H, CH), 4.23 (q, J = 6 Hz, 2H, ester-CH2), 4.59
(s, 1H, OH, D2O-exchangable), 5.85 (s, 1H, C4-H), 6.73–7.02 (m, 5H,
C6H5), 8.03 (brs, 1H, NH, D2O-exchangable). M.S. (EI): m/z (%): 492
(M+−1, 59), 420 (M+−CO2Et, 40), 401 (34), 271 (C19H27O, 76), 92
(100), 77 (36). Calc. for C31H43NO4 (493.677): C, 75.42; H, 8.78; N,
2.84; found: C, 75.18; H, 8.57; N, 2.57%.
2.1.2.3. 17-[Ethyl 3ꢀ-hydroxy-2ꢀ-(p-chlorophenylamino)butanoate]
androst-4-en-3-one (5b). Yellow crystals from absolute ethanol,
yield 0.76 g (72%), mp 65–67 ◦C, IR (KBr, cm−1): ꢀ = 3548–3380
(OH, NH), 3038 (CH-aromatic), 2978–2872 (CH-aliphatic), 1711
(C-3, C O), 1740 (CO-ester), 1643 (C C). 1H NMR (CDCl3, ppm):
ı = 0.99 (s, 3H, CH3-19), 1.07 (s, 3H, CH3-18), 1.32 (s, 3H, CH3),
1.38 (t, J = 6 Hz, 3H, ester-CH3), 3.58 (s, 1H, CH), 4.25 (q, J = 6 Hz,
2H, ester-CH2), 4.68 (s, 1H, OH, D2O-exchangable), 5.85 (s, 1H,
C4-H), 6.42 (dd, 2H-aromatic, JHH = 8 Hz), 7.05 (dd, 2H-aromatic,
JHH = 8 Hz), 8.12 (brs, 1H, NH, D2O-exchangable). M.S. (EI): m/z (%):
528 (M+, 39), 454 (M+−CO2Et, 56), 271 (C19H27O, 43), 126 (100),
111 (38). Calc. for C31H42 ClNO4 (528.122): C, 70.50; H, 8.02; N,
2.65; found: C, 70.74; H, 7.82; N, 2.91%.
2.1.1.2. Ethyl
17-(2ꢀ-methyloxiran-2ꢀ-yl)-3-oxo-androst-4-en-3ꢀ-
carboxylate (3). Dark yellow crystals from absolute ethanol,
yield 0.28 g (72%), mp 98–100 ◦C, IR (KBr, cm−1): ꢀ = 2956–2878
(CH-aliphatic), 1708 (C-3, C O), 1742 (ester-C O), 1643 (C C),
1H NMR (DMSO-d6, ppm): ı = 0.87 (s, 3H, CH3-19), 1.03 (s, 3H,
CH3-18), 1.28 (s, 3H, CH3), 1.37 (t, J = 7 Hz, 3H, ester-CH3), 3.45 (s,
1H, oxarine-CH), 4.25 (q, J = 7 Hz, 2H, ester-CH2), 5.76 (s, 1H, C4-H).
M.S. (EI): m/z (%): 400 (M+, 29), 327 (M+−CO2Et, 45), 271 (C19H27O,
100), 129 (40), 74 (67), 57 (38), Calc. for C25H36O4 (400.551): C,
74.96; H, 9.06; found: C, 75.08; H, 9.29%.
2.1.1.3. 17-(3ꢀ-Acetyl-2ꢀ-methyloxiran-2ꢀ-yl)androst-4-en-3-one
(15). Pale yellow crystals from dioxane, yield 0.29 g (78%), mp
192–194 ◦C, IR (KBr, cm−1): ꢀ = 2960–2875 (CH-aliphatic), 1708
(C-3, C O), 1718 (COCH3), 1645 (C C), 1H NMR (DMSO-d6, ppm):
ı = 0.81 (s, 3H, CH3-19), 1.05 (s, 3H, CH3-18), 1.32 (s, 3H, CH3), 2.07
(s, 3H, COCH3), 3.37 (s, 1H, oxarine-CH), 5.87 (s, 1H, C4-H). M.S.
(EI): m/z (%): 370 (M+, 43), 327 (M+−COCH3, 38), 271 (C19H27O,
2.1.2.4. 17-[Ethyl3ꢀ-hydroxy-2ꢀ-(p-methoxyphenylamino)butanoate]
androst-4-en-3-one (5c). Brown crystals from absolute ethanol,
yield 0.80 g (77%), mp 175–177 ◦C, IR (KBr, cm−1): ꢀ = 3546–3380
(OH, NH), 3035 (CH-aromatic), 2978–2876 (CH-aliphatic), 1705
(C-3, C O), 1745 (CO-ester), 1647 (C C). 1H NMR (CDCl3, ppm):
ı = 1.02 (s, 3H, CH3-19), 1.12 (s, 3H, CH3-18), 1.28 (s, 3H, CH3), 1.35

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R.M. Mohareb et al. / Steroids 76 (2011) 1190–1203
(t, J = 5.3 Hz, 3H, ester-CH3), 3.49 (s, 1H, CH), 3.78 (s, 3H, OCH3), 4.25
(q, J = 5.3 Hz, 2H, ester-CH2), 4.65 (s, 1H, OH, D2O-exchangable),
5.80 (s, 1H, C4-H), 6.38 (dd, 2H-aromatic, JHH = 8 Hz), 7.02 (dd,
2H-aromatic, JHH = 8 Hz), 8.20 (brs, 1H, NH, D2O-exchangable).
M.S. (EI): m/z (%): 521 (M+−2, 25), 450 (M+−CO2Et, 64), 401 (37),
271 (C19H27O, 50), 122 (100), 107 (36). 13C NMR (CDCl3, ppm):
ı = 35.2 (C-1), 34.3, (C-2), 198.6 (C-3), 124.0 (C-4), 171.5 (C-5), 32.7
(C-6), 31.0 (C-7), 36.3 (C-8), 50.0 (C-9), 37.2 (C-10), 23.1 (C-11),
37.5 (C-12), 38.7 (C-13), 56.2 (C-14), 27.3 (C-15), 21.5 (C-16),
56.3 (C-17), 23.0 (C-18), 21.7 (C-19), 71.9 (CH), 21.4 (CH3), 171.0
(CO2Et), 62.3 (CO2CH2CH3), 14.5 (CH2CH3), 139.2, 115.6, 114.0,
150.4 (C-phenyl). Calc. for C32H45NO5 (523.703): C, 73.39; H, 8.66;
N, 2.67; found: C, 73.63; H, 8.39; N, 2.83%.
(100), 105 (60), 92 (33). Calc. for C35H43NO3 (525.721): C, 79.96;
H, 8.24; N, 2.66; found: C, 80.23; H, 7.98; N, 2.46%.
2.1.2.9. 17-[2ꢀ-Hydroxy-4ꢀ-oxo-4ꢀ-phenyl-3ꢀ-(p-
chlorophenylamino)butan-2-yl]androst-4-en-3-one (25b). Brown
crystals from absolute ethanol, yield 0.85 g (76%), mp 189–190 ◦C,
IR (KBr, cm−1): ꢀ = 3540–3378 (OH, NH), 3035 (CH-aromatic),
2978–2870 (CH-aliphatic), 1716 (C-3, C O), 1726 (COPh), 1648
(C C). 1H NMR (DMSO-d6, ppm): ı = 0.98 (s, 3H, CH3-19), 1.10
(s, 3H, CH3-18), 1.32 (s, 3H, CH3), 4.12 (s, 1H, CH), 4.68 (s, 1H,
OH, D2O-exchangable), 5.75 (s, 1H, C4-H), 6.37–7.85 (m, 9H,
aromatic-H), 8.35 (brs, 1H, NH, D2O-exchangable). M.S. (EI): m/z
(%): 559 (M+, 34), 453 (M+−COPh, 14), 288 (53), 271 (C19H27O, 76),
126 (100), 105 (60). Calc. for C35H42ClNO3 (559.285): C, 75.04; H,
7.56; N, 2.50 found: C, 75.26; H, 7.80; N, 2.23%.
2.1.2.5. 17-[2ꢀ-Hydroxy-4ꢀ-oxo-4ꢀ-methyl-3ꢀ-(phenylamino)butan-
2-yl]androst-4-en-3-one (16a). Yellow crystals from methanol,
yield 0.72 g (78%), mp 192–194 ◦C, IR (KBr, cm−1): ꢀ = 3548–3390
(OH, NH), 3030 (CH-aromatic), 2973–2878 (CH-aliphatic), 1710
(C-3, C O), 1722 (COCH3), 1640 (C C). 1H NMR (CDCl3, ppm):
ı = 0.96 (s, 3H, CH3-19), 1.13 (s, 3H, CH3-18), 1.30 (s, 3H, CH3), 2.10
(s, 3H, COCH3), 3.55 (s, 1H, CH), 4.52 (s, 1H, OH, D2O-exchangable),
5.76 (s, 1H, C4-H), 6.78–7.06 (m, 5H, C6H5), 8.53 (brs, 1H, NH, D2O-
exchangable). M.S. (EI): m/z (%): 463 (M+, 39), 420 (M+−COCH3,
47), 371 (23), 315 (100), 271 (C19H27O, 64), 92 (62), 77 (45). Calc.
for C30H41NO3 (463.651): C, 77.71; H, 8.91; N, 3.02; found: C,
77.96; H, 8.76; N, 2.83%.
2.1.2.10. 17-[2ꢀ-Hydroxy-4ꢀ-oxo-4ꢀ-phenyl-3ꢀ-(p-
methoxyphenylamino)butan-2-yl]androst-4-en-3-one
(25c). Pale
yellow powder from methanol, yield 0.82 g (74%), mp 123–125 ◦C,
IR (KBr, cm−1): ꢀ = 3536–3382 (OH, NH), 3032 (CH-aromatic),
2982–2876 (CH-aliphatic), 1698 (C-3, C O), 1724 (COPh), 1652
(C C). 1H NMR (DMSO-d6, ppm): ı = 1.03 (s, 3H, CH3-19), 1.14 (s,
3H, CH3-18), 1.33 (s, 3H, CH3), 3.80 (s, 3H, OCH3), 4.15 (s, 1H, CH),
4.72 (s, 1H, OH, D2O-exchangable), 5.80 (s, 1H, C4-H), 6.40–7.86
(m, 9H, aromatic-H), 8.35 (brs, 1H, NH, D2O-exchangable). M.S.
(EI): m/z (%): 554 (M+−1, 53), 450 (M+−COPh, 64), 284 (30), 271
(C19H27O, 63), 122 (100), 105 (60). Calc. for C36H45NO4 (555.747):
C, 77.80; H, 8.16; N, 2.52; found: C, 78.02; H, 7.97; N, 2.30%.
2.1.2.6. 17-[2ꢀ-Hydroxy-4ꢀ-oxo-4ꢀ-methyl-3ꢀ-(p-
chlorophenylamino)butan-2-yl]androst-4-en-3-one
(16b). Pale
2.1.3. Synthesis of 17-(pyrazol-5ꢀ-yl)androst-4-ene-3-one
brown crystals from absolute ethanol, yield 0.74 g (75%), mp
derivatives (8a,b), (9a,b), (10a,b), (18a,b), (19a,b), (20a,b),
(27a,b), (28a,b) and (29a,b)
110–112 ◦C, IR (KBr, cm−1): ꢀ = 3546–3378 (OH, NH), 3028 (CH-
aromatic), 2978–2872 (CH-aliphatic), 1706 (C-3,
C
O), 1726
2.1.3.1. General procedure. To a solution of either compound 5a
(0.98 g, 2 mmol), 5b (1.05 g, 2 mmol), 5c (1.04 g, 2 mmol), 16a
(0.92 g, 2 mmol), 16b (0.99 g, 2 mmol), 16c (0.98 g, 2 mmol) 25a
(1.05 g, 2 mmol), 25b (1.12 g, 2 mmol) or 25c (1.11 g, 2 mmol) in
absolute ethanol (20 mL) containing a catalytic amount of triethy-
lamine (1 mL) either hydrazine hydrate 6a (0.1 g, 2 mmol) or phenyl
hydrazine 6b (0.21 g, 2 mmol) was added. The reaction mixture, in
each case, was heated under reflux for 8–12 h until all the reactants
had disappeared as indicated by TLC. The reaction mixture poured
over an ice/water mixture and neutralized with dilute hydrochlo-
ric acid. The resulted semisolid was subjected to extraction with
chloroform (2–3 × 30 mL). The organic layer was dried over mag-
nesium sulfate and then filtered. The solid product that formed in
each case on removal of the solvent in vacuum was collected, dried
and crystallized from the appropriate solvent.
(COCH3), 1643 (C C). 1H NMR (DMSO-d6, ppm): ı = 0.98 (s, 3H,
CH3-19), 1.09 (s, 3H, CH3-18), 1.32 (s, 3H, CH3), 2.10 (s, 3H, COCH3),
3.57 (s, 1H, CH), 4.64 (s, 1H, OH, D2O-exchangable), 5.75 (s, 1H,
C4-H), 6.49 (dd, 2H-aromatic, JHH = 8 Hz), 7.15 (dd, 2H-aromatic,
JHH = 8 Hz), 8.35 (brs, 1H, NH, D2O-exchangable). M.S. (EI): m/z (%):
497 (M+−1, 35), 454 (M+−COCH3, 24), 371 (34), 271 (C19H27O, 76),
126 (100), 111 (22). Calc. for C30H40ClNO3 (498.046): C, 72.34; H,
8.09; N, 2.81; found: C, 72.14; H, 7.87; N, 2.98%.
2.1.2.7. 17-[2ꢀ-Hydroxy-4ꢀ-oxo-4ꢀ-methyl-3ꢀ-(p-
methoxyphenylamino)butan-2-yl]androst-4-en-3-one
(16c).
Reddish brown powder from ethanol (70%), yield 0.78 g (79%),
mp 136–137 ◦C, IR (KBr, cm−1): ꢀ = 3538–3387 (OH, NH), 3033
(CH-aromatic), 2980–2876 (CH-aliphatic), 1703 (C-3, C O), 1727
(COCH3), 1645 (C C). 1H NMR (DMSO-d6, ppm): ı = 1.02 (s, 3H, CH3-
19), 1.12 (s, 3H, CH3-18), 1.37 (s, 3H, CH3), 2.09 (s, 3H, COCH3), 3.48
(s, 1H, CH), 3.73 (s, 3H, OCH3), 4.78 (s, 1H, OH, D2O-exchangable),
5.83 (s, 1H, C4-H), 6.37 (dd, 2H-aromatic, JHH = 8 Hz), 7.05 (dd,
2H-aromatic, JHH = 8 Hz), 8.30 (brs, 1H, NH, D2O-exchangable). M.S.
(EI): m/z (%): 493 (M+, 46), 450 (M+−COCH3, 32), 371 (100), 271
(C19H27O, 27), 122 (47), 107 (18). Calc. for C31H43NO4 (493.677):
C, 75.42; H, 8.78; N, 2.84; found: C, 75.20; H, 9.02; N, 2.63%.
2.1.3.2. 17-(4ꢀ,5ꢀ-Dihydro-3ꢀ-hydroxy-5ꢀ-methyl-4ꢀ-phenylamino-
1ꢀH-pyrazol-5ꢀ-yl)androst-4-ene-3-one (8a). Red crystals from
absolute ethanol, yield 0.75 g (82%), mp 290–292 ◦C, IR (KBr,
cm−1): ꢀ = 3542–3378 (OH, 2NH), 3030 (CH-aromatic), 2978–2869
(CH-aliphatic), 1703 (C-3, C O), 1670 (C N), 1646 (C C). 1H NMR
(DMSO-d6, ppm): ı = 1.07 (s, 3H, CH3-19), 1.12 (s, 3H, CH3-18),
1.27 (s, 3H, CH3), 4.68 (s, 1H, OH, D2O-exchangable), 5.83 (s,
1H, C4-H), 6.83–7.06 (m, 5H, C6H5), 8.34, 9.05 (2brs, 2H, 2NH,
D2O-exchangable). M.S. (EI): m/z (%): 461 (M+, 37), 271 (C19H27O,
100), 266 (23), 92 (60), 77 (84). Calc. for C29H39N3O2 (461.639): C,
75.45; H, 8.52; N, 9.10; found: C, 75.24; H, 8.30; N, 8.83%.
2.1.2.8. 17-[2ꢀ-Hydroxy-4ꢀ-oxo-4ꢀ-phenyl-3ꢀ-(phenylamino)butan-
2-yl]androst-4-en-3-one (25a). Yellow powder from dioxane, yield
0.86 g (82%), mp 178–180 ◦C, IR (KBr, cm−1): ꢀ = 3534–3397 (OH,
NH), 3030 (CH-aromatic), 2965–2860 (CH-aliphatic), 1706 (C-3,
2.1.3.3. 17-(4ꢀ,5ꢀ-Dihydro-3ꢀ-hydroxy-5ꢀ-methyl-4ꢀ-phenylamino-
1ꢀ-phenylpyrazol-5ꢀ-yl)androst-4-ene-3-one (8b). Yellowish brown
powder from ethanol (70%), yield 0.83 g (78%), mp 145–147 ◦C,
IR (KBr, cm−1): ꢀ = 3538–3385 (OH, NH), 3025 (CH-aromatic),
2978–2872 (CH-aliphatic), 1705 (C-3, C O), 1664 (C N), 1642
(C C). 1H NMR (DMSO-d6, ppm): ı = 0.97 (s, 3H, CH3-19), 1.12 (s,
C
O), 1728 (COPh), 1643 (C C). 1H NMR (CDCl3, ppm): ı = 1.06 (s,
3H, CH3-19), 1.13 (s, 3H, CH3-18), 1.28 (s, 3H, CH3), 4.17 (s, 1H, CH),
4.58 (s, 1H, OH, D2O-exchangable), 5.87 (s, 1H, C4-H), 6.48–7.86
(m, 10H, 2C6H5), 8.85 (brs, 1H, NH, D2O-exchangable). M.S. (EI):
m/z (%): 525 (M+, 42), 420 (M+−COPh, 14), 271 (C19H27O, 76), 254

R.M. Mohareb et al. / Steroids 76 (2011) 1190–1203
1193
3H, CH3-18), 1.35 (s, 3H, CH3), 4.87 (s, 1H, OH, D2O-exchangable),
5.74 (s, 1H, C4-H), 6.65–7.15 (m, 10H, aromatic-H), 8.25 (brs, 1H,
NH, D2O-exchangable). 13C NMR (DMSO-d6, ppm): ı = 35.3 (C-1),
34.0, (C-2), 198.5 (C-3), 124.3 (C-4), 170.0 (C-5), 32.7 (C-6), 31.6
(C-7), 35.4 (C-8), 50.6 (C-9), 37.3 (C-10), 22.6 (C-11), 37.5 (C-12),
38.4 (C-13), 56.0 (C-14), 27.6 (C-15), 23.0 (C-16), 46.8 (C-17), 22.7
(C-18), 20.3 (C-19), 19.2 (CH3), 156.2, 73.4, 53.0 (C-pyrazole),
147.0, 143.6, 129.0, 117.2, 113.6, (C-phenyl). M.S. (EI): m/z (%): 537
(M+, 64), 271 (C19H27O, 100), 266 (34), 92 (56), 77 (67). Calc. for
Calc. for C36H45N3O3 (567.761): C, 76.16; H, 7.99; N, 7.40; found:
C, 76.37; H, 8.18; N, 7.18.
2.1.3.8. 17-(4ꢀ,5ꢀ-Dihydro-3ꢀ,5ꢀ-dimethyl-4ꢀ-phenylamino-1ꢀH-
pyrazol-5ꢀ-yl)androst-4-ene-3-one (18a). Pale red crystals from
absolute ethanol, yield 0.75 g (82%), mp 229–230 ◦C, IR (KBr,
cm−1): ꢀ = 3405–3378 (2NH), 3030 (CH-aromatic), 2978–2869
(CH-aliphatic), 1717 (C-3, C O), 1672 (C N), 1646 (C C). 1H NMR
(DMSO-d6, ppm): ı = 1.07 (s, 3H, CH3-19), 1.16 (s, 3H, CH3-18),
1.31 (s, 6H, 2CH3), 5.80 (s, 1H, C4-H), 6.45–7.08 (m, 5H, C6H5), 8.45,
9.35 (2brs, 2H, 2NH, D2O-exchangable). M.S. (EI): m/z (%): 459
(M+, 26), 271 (C19H27O, 100), 92 (47), 77 (55). Calc. for C30H41N3O
(459.666): C, 78.39; H, 8.99; N, 9.14; found: C, 78.60; H, 9.18; N,
8.93.
C35H43N3O2 (537.735): C, 78.18; H, 8.06; N, 7.81; found: C, 77.93;
H, 8.23; N, 8.02.
2.1.3.4. 17-[4ꢀ,5ꢀ-Dihydro-3ꢀ-hydroxy-5ꢀ-methyl-4ꢀ-(p-
chlorophenylamino)-1ꢀH-pyrazol-5ꢀ-yl]androst-4-ene-3-one
(9a).
Brown crystals from dioxane, yield 0.78 g (75%), mp 268–270 ◦C,
IR (KBr, cm−1): ꢀ = 3487–3365 (OH, 2NH), 3028 (CH-aromatic),
2969–2875 (CH-aliphatic), 1713 (C-3, C O), 1665 (C N), 1645
(C C). 1H NMR (DMSO-d6, ppm): ı = 1.12 (s, 3H, CH3-19), 1.18 (s,
3H, CH3-18), 1.26 (s, 3H, CH3), 4.57 (s, 1H, OH, D2O-exchangable),
5.80 (s, 1H, C4-H), 6.37 (dd, 2H-aromatic, JHH = 8 Hz), 7.12 (dd, 2H-
aromatic, JHH = 8 Hz), 8.42, 9.45 (2brs, 2H, 2NH, D2O-exchangable).
M.S. (EI): m/z (%): 496 (M+, 37), 271 (C19H27O, 45), 126 (35) 111
(84), 99 (100). Calc. for C29H38ClN3O2 (496.084): C, 70.21; H, 7.72;
N, 8.47; found: C, 70.45; H, 7.98; N, 8.73.
2.1.3.9. 17-(4ꢀ,5ꢀ-Dihydro-3ꢀ,5ꢀ-dimethyl-4ꢀ-phenylamino-1ꢀ-
phenylpyrazol-5ꢀ-yl)androst-4-ene-3-one (18b). Yellow powder
from dioxane, yield 0.72 g (68%), mp 135–137 ◦C, IR (KBr, cm−1):
ꢀ = 3385 (NH), 3035 (CH-aromatic), 2967–2872 (CH-aliphatic),
1715 (C-3, C O), 1668 (C N), 1642 (C C). 1H NMR (DMSO-d6,
ppm): ı = 1.02 (s, 3H, CH3-19), 1.15 (s, 3H, CH3-18), 1.35 (s, 6H,
2CH3), 5.82 (s, 1H, C4-H), 6.43–7.10 (m, 10H, aromatic-H), 8.46 (brs,
1H, NH, D2O-exchangable). 13C NMR (DMSO-d6, ppm): ı = 36.3
(C-1), 33.7, (C-2), 198.7 (C-3), 127.3 (C-4), 171.3 (C-5), 34.7 (C-6),
31.6 (C-7), 35.7 (C-8), 52.6 (C-9), 37.3 (C-10), 22.4 (C-11), 37.5
(C-12), 38.7 (C-13), 56.4 (C-14), 27.6 (C-15), 23.0 (C-16), 46.8
(C-17), 23.2 (C-18), 21.3 (C-19), 19.2, 16.7 (2CH3), 155.2, 70.4, 56.0
(C-pyrazole), 147.0, 143.1, 128.2, 117.2, 113.6, (C-phenyl). M.S.
(EI): m/z (%): 535 (M+, 64), 271 (C19H27O, 39), 264 (34), 92 (29),
77 (26). Calc. for C36H45N3O (535.762): C, 80.70; H, 8.47; N, 7.84;
found: C, 80.93; H, 8.29; N, 8.05.
2.1.3.5. 17-[4ꢀ,5ꢀ-Dihydro-3ꢀ-hydroxy-5ꢀ-methyl-4ꢀ-(p-
chlorophenylamino)-1ꢀ-phenyl-pyrazol-5ꢀ-yl]androst-4-ene-3-one
(9b). Yellow crystals from absolute ethanol, yield 0.82 g (72%),
mp 198–200 ◦C, IR (KBr, cm−1): ꢀ = 3532–3372 (OH, NH), 3025
(CH-aromatic), 2974–2860 (CH-aliphatic), 1715 (C-3, C O), 1667
(C N), 1642 (C C). 1H NMR (DMSO-d6, ppm): ı = 1.03 (s, 3H,
CH3-19), 1.14 (s, 3H, CH3-18), 1.30 (s, 3H, CH3), 4.65 (s, 1H, OH,
D2O-exchangable), 5.74 (s, 1H, C4-H), 6.34 (dd, 2H-aromatic,
JHH = 7 Hz), 6.45–7.04 (m, 5H, C6H5), 7.14 (dd, 2H-aromatic,
JHH = 7 Hz), 8.37 (brs, 1H, NH, D2O-exchangable). M.S. (EI): m/z (%):
573 (M++1, 37), 271 (C19H27O, 60), 126 (34), 111 (50), 77 (67). Calc.
for C35H42ClN3O2 (572.180): C, 73.47; H, 7.40; N, 7.34; found: C,
73.62; H, 7.59; N, 7.64.
2.1.3.10. 17-[4ꢀ,5ꢀ-Dihydro-3ꢀ,5ꢀ-dimethyl-4ꢀ-(p-
chlorophenylamino)-1ꢀH-pyrazol-5ꢀ-yl]androst-4-ene-3-one (19a).
Brown crystals from methanol, yield 0.73 g (74%), mp 163–165 ◦C,
IR (KBr, cm−1): ꢀ = 3427–3365 (2NH), 3028 (CH-aromatic),
2969–2875 (CH-aliphatic), 1713 (C-3, C O), 1665 (C N), 1645
(C C). 1H NMR (DMSO-d6, ppm): ı = 1.02 (s, 3H, CH3-19), 1.17
(s, 3H, CH3-18), 1.30 (s, 6H, 2CH3), 5.80 (s, 1H, C4-H), 6.56 (dd,
2H-aromatic, JHH = 8 Hz), 7.06 (dd, 2H-aromatic, JHH = 8 Hz), 8.56,
9.47 (2brs, 2H, 2NH, D2O-exchangable). M.S. (EI): m/z (%): 494 (M+,
72), 271 (C19H27O, 53), 126 (100) 111 (24). Calc. for C30H40ClN3O
(494.111): C, 72.92; H, 8.16; N, 8.50; found: C, 73.11; H, 7.96; N,
8.71.
2.1.3.6. 17-[4ꢀ,5ꢀ-Dihydro-3ꢀ-hydroxy-5ꢀ-methyl-4ꢀ-(p-
methoxyphenylamino)-1ꢀH-pyrazol-5ꢀ-yl]androst-4-ene-3-one
(10a). Brown crystals from methanol, yield 0.73 g (75%), mp
290–292 ◦C, IR (KBr, cm−1): ꢀ = 3495–3375 (OH, 2NH), 3028
(CH-aromatic), 2969–2875 (CH-aliphatic), 1713 (C-3, C O), 1665
(C N), 1645 (C C). 1H NMR (DMSO-d6, ppm): ı = 1.02 (s, 3H,
CH3-19), 1.17 (s, 3H, CH3-18), 1.25 (s, 3H, CH3), 3.78 (s, 3H, OCH3),
4.62 (s, 1H, OH, D2O-exchangable), 5.87 (s, 1H, C4-H), 6.52 (dd,
2H-aromatic, JHH = 8 Hz), 6.72 (dd, 2H-aromatic, JHH = 8 Hz), 8.42,
9.38 (2brs, 2H, 2NH, D2O-exchangable). M.S. (EI): m/z (%): 491
(M+, 37), 271 (C19H27O, 100), 202 (65), 122 (39), 107 (28). Calc. for
2.1.3.11. 17-[4ꢀ,5ꢀ-Dihydro-3ꢀ,5ꢀ-dimethyl-4ꢀ-(p-
chlorophenylamino)-1ꢀ-phenylpyrazol-5ꢀ-yl]androst-4-ene-3-one
(19b). Yellow crystals from absolute ethanol, yield 0.94 g (83%),
mp 141–143 ◦C, IR (KBr, cm−1): ꢀ = 3372 (NH), 3035 (CH-aromatic),
2970–2864 (CH-aliphatic), 1713 (C-3, C O), 1668 (C N), 1625
(C C). 1H NMR (DMSO-d6, ppm): ı = 0.97 (s, 3H, CH3-19), 1.12
(s, 3H, CH3-18), 1.32 (s, 6H, 2CH3), 5.74 (s, 1H, C4-H), 6.37 (dd,
2H-aromatic, JHH = 7 Hz), 6.45–7.04 (m, 5H, C6H5), 7.08 (dd, 2H-
aromatic, JHH = 7 Hz), 8.47 (brs, 1H, NH, D2O-exchangable). 13C
NMR (CDCl3, ppm): ı = 35.3 (C-1), 34.2, (C-2), 198.0 (C-3), 124.5
(C-4), 171.2 (C-5), 32.7 (C-6), 31.0 (C-7), 35.4 (C-8), 50.6 (C-9),
37.8 (C-10), 22.7 (C-11), 37.2 (C-12), 38.4 (C-13), 56.6 (C-14), 27.6
(C-15), 22.4 (C-16), 47.4 (C-17), 22.7 (C-18), 20.3 (C-19), 16.7, 19.3
(2CH3), 155.0, 73.8, 56.0 (C-pyrazole), 114.3, 129.7, 122,0, 143.3,
128.6, 117.2 (C-phenyl). M.S. (EI): m/z (%): 572 (M++2, 52), 271
(C19H27O, 23), 126 (100), 77 (24). Calc. for C36H44ClN3O (570.207):
C, 75.83; H, 7.78; N, 7.37; found: C, 76.03; H, 7.59; N, 7.60.
C30H41N3O3 (491.665): C, 73.29; H, 8.41; N, 8.55; found: C, 73.08;
H, 8.59; N, 8.70.
2.1.3.7. 17-[4ꢀ,5ꢀ-Dihydro-3ꢀ-hydroxy-5ꢀ-methyl-4ꢀ-(p-
methoxyphenylamino)-1ꢀ-phenylpyrazol-5ꢀ-yl]androst-4-ene-3-one
(10b). Pale brown powder from absolute ethanol, yield 0.88 g
(78%), mp 249–251 ◦C, IR (KBr, cm−1): ꢀ = 3520–3386 (OH, NH),
3032 (CH-aromatic), 2974–2860 (CH-aliphatic), 1710 (C-3, C O),
1667 (C N), 1640 (C C). 1H NMR (DMSO-d6, ppm): ı = 1.05 (s,
3H, CH3-19), 1.14 (s, 3H, CH3-18), 1.30 (s, 3H, CH3), 3.75 (s, 3H,
OCH3), 4.35 (s, 1H, OH, D2O-exchangable), 5.80 (s, 1H, C4-H), 6.48
(dd, 2H-aromatic, JHH = 8 Hz), 6.64 (dd, 2H-aromatic, JHH = 8 Hz),
6.87–7.06 (m, 5H, C6H5), 8.37 (brs, 1H, NH, D2O-exchangable). M.S.
(EI): m/z (%): 567 (M+, 43), 271 (C19H27O, 63), 122 (58), 107 (100).
2.1.3.12. 17-[4ꢀ,5ꢀ-Dihydro-3ꢀ,5ꢀ-dimethyl-4ꢀ-(p-
methoxyphenylamino)-1ꢀH-pyrazol-5ꢀ-yl]androst-4-ene-3-one
(20a). Yellow crystals from absolute ethanol, yield 0.76 g (78%),

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R.M. Mohareb et al. / Steroids 76 (2011) 1190–1203
2.1.3.17. 17-[4ꢀ,5ꢀ-Dihydro-1,3ꢀ-diphenyl-5ꢀ-methyl-4ꢀ-(p-
chlorophenylamino)pyrazol-5ꢀ-yl]androst-4-ene-3-one
mp 192–194 ◦C, IR (KBr, cm−1): ꢀ = 3395–3364 (2NH), 3030 (CH-
aromatic), 2969–2875 (CH-aliphatic), 1715 (C-3, C O), 1660 (C N),
1595 (C C). 1H NMR (DMSO-d6, ppm): ı = 1.02 (s, 3H, CH3-19), 1.14
(s, 3H, CH3-18), 1.28 (s, 6H, 2CH3), 3.78 (s, 3H, OCH3), 5.87 (s, 1H,
C4-H), 6.52 (dd, 2H-aromatic, JHH = 8 Hz), 6.72 (dd, 2H-aromatic,
JHH = 8 Hz), 8.52, 9.08 (2brs, 2H, 2NH, D2O-exchangable). M.S. (EI):
m/z (%): 450 (M++1, 57), 271 (C19H27O, 100), 122 (39), 107 (58).
Calc. for C31H43N3O2 (489.692): C, 76.03; H, 8.85; N, 8.58; found:
C, 75.82; H, 9.02; N, 8.74.
(28b).
Yellow crystals from absolute ethanol, yield 0.85 g (68%), mp
241–243 ◦C, IR (KBr, cm−1): ꢀ = 3372 (NH), 3035 (CH-aromatic),
2970–2864 (CH-aliphatic), 1703 (C-3, C O), 1660 (C N), 1580
(C C). 1H NMR (DMSO-d6, ppm): ı = 0.98 (s, 3H, CH3-19), 1.12 (s,
3H, CH3-18), 1.32 (s, 3H, CH3), 5.82 (s, 1H, C4-H), 6.35 (dd, 2H-
aromatic, JHH = 7 Hz), 7.05 (dd, 2H-aromatic, JHH = 7 Hz), 7.30–7.64
(m, 10H, 2C6H5), 8.87 (brs, 1H, NH, D2O-exchangable). M.S. (EI):
m/z (%): 632 (M+, 46), 271 (C19H27O, 23), 126 (100), 77 (34). Calc.
for C41H46ClN3O (632.276): C, 77.88; H, 7.33; N, 6.65; found: C,
78.12; H, 7.59; N, 6.83.
2.1.3.13. 17-[4ꢀ,5ꢀ-Dihydro-3ꢀ,5ꢀ-dimethyl-4ꢀ-(p-
methoxyphenylamino)-1ꢀ-phenylpyrazol-5ꢀ-yl]androst-4-ene-3-one
(20b). Orange crystals from DMF, yield 0.90 g (80%), mp
203–205 ◦C, IR (KBr, cm−1): ꢀ = 3387 (NH), 3032 (CH-aromatic),
2973–2876 (CH-aliphatic), 1715 (C-3, C O), 1658 (C N), 1598
(C C). 1H NMR (DMSO-d6, ppm): ı = 0.96 (s, 3H, CH3-19), 1.12 (s,
3H, CH3-18), 1.32 (s, 6H, 2CH3), 3.85 (s, 3H, OCH3), 5.80 (s, 1H, C4-
H), 6.32 (dd, 2H-aromatic, JHH = 8 Hz), 6.43–7.08 (m, 5H, C6H5), 7.12
(dd, 2H-aromatic, JHH = 8 Hz), 9.28 (brs, 1H, NH, D2O-exchangable).
M.S. (EI): m/z (%): 566 (M++1, 57), 271 (C19H27O, 100), 122 (39),
107 (58), 77 (27). Calc. for C37H47N3O2 (565.788): C, 78.54; H, 8.37;
N, 7.43; found: C, 78.32; H, 8.59; N, 7.68.
2.1.3.18. 17-[4ꢀ,5ꢀ-Dihydro-5ꢀ-methyl-4ꢀ-(p-methoxyphenylamino)-
3ꢀ-phenyl-1ꢀH-pyrazol-5ꢀ-yl]androst-4-ene-3-one
(29a). Orange
crystals from dioxane, yield 0.85 g (78%), mp 198–199 ◦C, IR (KBr,
cm−1): ꢀ = 3435–3327 (2NH), 3033 (CH-aromatic), 2969–2875
(CH-aliphatic), 1710 (C-3, C O), 1656 (C N), 1578 (C C). 1H NMR
(DMSO-d6, ppm): ı = 1.02 (s, 3H, CH3-19), 1.15 (s, 3H, CH3-18), 1.30
(s, 3H, CH3), 3.76 (s, 3H, OCH3), 5.87 (s, 1H, C4-H), 6.47 (dd, 2H-
aromatic, JHH = 8 Hz), 6.87 (dd, 2H-aromatic, JHH = 8 Hz), 7.23–7.48
(m, 5H, C6H5), 8.45, 9.18 (2brs, 2H, 2NH, D2O-exchangable). M.S.
(EI): m/z (%): 552 (M++1, 53), 271 (C19H27O, 100), 122 (39), 107
(58). Calc. for C36H45N3O2 (551.761): C, 78.36; H, 8.22; N, 7.62;
found: C, 78.13; H, 7.99; N, 7.85.
2.1.3.14. 17-(4ꢀ,5ꢀ-Dihydro-5ꢀ-methyl-4ꢀ-phenylamino-3ꢀ-phenyl-
1ꢀH-pyrazol-5ꢀ-yl)androst-4-ene-3-one (27a). Yellow crystals from
absolute ethanol, yield 0.75 g (72%), mp 200–202 ◦C, IR (KBr,
cm−1): ꢀ = 3415–3378 (2NH), 3030 (CH-aromatic), 2978–2869
(CH-aliphatic), 1714 (C-3, C O), 1667 (C N), 1626 (C C). 1H NMR
(DMSO-d6, ppm): ı = 1.07 (s, 3H, CH3-19), 1.16 (s, 3H, CH3-18),
1.30 (s, 3H, CH3), 5.80 (s, 1H, C4-H), 6.65–7.28 (m, 10H, 2C6H5),
8.35, 9.05 (2brs, 2H, 2NH, D2O-exchangable). M.S. (EI): m/z (%): 522
(M++1, 46), 271 (C19H27O, 73), 92 (47), 77 (55). Calc. for C35H43N3O
(521.735): C, 80.57; H, 8.31; N, 8.05; found: C, 80.33; H, 8.49; N,
8.29.
2.1.3.19. 17-[4ꢀ,5ꢀ-Dihydro-1ꢀ,3ꢀ-diphenyl-5ꢀ-methyl-4ꢀ-(p-
methoxyphenylamino)pyrazol-5ꢀ-yl]androst-4-ene-3-one
(29b).
Yellowish brown powder from ethanol (70%), yield 1.04 g (83%),
mp 123–125 ◦C, IR (KBr, cm−1): ꢀ = 3386 (NH), 3035 (CH-aromatic),
2973–2876 (CH-aliphatic), 1708 (C-3, C O), 1662 (C N), 1596
(C C). 1H NMR (DMSO-d6, ppm): ı = 0.96 (s, 3H, CH3-19), 1.12
(s, 3H, CH3-18), 1.32 (s, 3H, CH3), 3.85 (s, 3H, OCH3), 5.80 (s, 1H,
C4-H), 6.43 (dd, 2H-aromatic, JHH = 8 Hz), 7.10 (dd, 2H-aromatic,
JHH = 8 Hz), 7.25–7.65 (m, 10H, 2C6H5), 8.68 (brs, 1H, NH, D2O-
exchangable). M.S. (EI): m/z (%): 627 (M+, 35), 271 (C19H27O, 67),
122 (100), 107 (58), 77 (27). Calc. for C42H49N3O2 (627.857) C,
80.34; H, 7.87; N, 6.69; found: C, 80.55; H, 7.69; N, 6.88.
2.1.3.15. 17-(4ꢀ,5ꢀ-Dihydro-1ꢀ,3ꢀ-diphenyl-5ꢀ-methyl-4ꢀ-
phenylaminopyrazol-5ꢀ-yl)androst-4-ene-3-one
(27b). Yellow
powder from dioxane, yield 1.00 g (84%), mp 136–137 ◦C, IR
(KBr, cm−1): ꢀ = 3385 (NH), 3030 (CH-aromatic), 2967–2872
(CH-aliphatic), 1713 (C-3, C O), 1668 (C N), 1622 (C C). 1H NMR
(DMSO-d6, ppm): ı = 1.02 (s, 3H, CH3-19), 1.15 (s, 3H, CH3-18), 1.37
(s, 3H, CH3), 5.82 (s, 1H, C4-H), 6.48–7.38 (m, 15H, aromatic-H),
8.76 (brs, 1H, NH, D2O-exchangable). 13C NMR (DMSO-d6, ppm):
ı = 36.3 (C-1), 33.7, (C-2), 198.7 (C-3), 127.3 (C-4), 171.3 (C-5), 34.7
(C-6), 32.5 (C-7), 35.7 (C-8), 52.8 (C-9), 38.0 (C-10), 22.6 (C-11),
37.5 (C-12), 38.9 (C-13), 56.7 (C-14), 27.6 (C-15), 23.3 (C-16),
46.8 (C-17), 23.2 (C-18), 21.8 (C-19), 19.2 (CH3), 155.2, 72.4, 56.3
(C-pyrazole), 147.0, 143.1, 128.7, 117.2, 113.8 (C-phenyl). M.S. (EI):
m/z (%): 597 (M+, 44), 271 (C19H27O, 100), 92 (29), 77 (26). Calc. for
2.1.4. Synthesis of 20-(imidazole-3-yl)pregn-4-ene-3-one
derivatives (13a–c), (22a–c), (31a–c)
2.1.4.1. General procedure. To a solution of either compound 5a
(0.98 g, 2 mmol), 5b (1.05 g, 2 mmol), 5c (1.04 g, 2 mmol), 16a
(0.92 g, 2 mmol), 16b (0.99 g, 2 mmol), 16c (0.98 g, 2 mmol) 25a
(1.05 g, 2 mmol), 25b (1.12 g, 2 mmol), or 25c (1.11 g, 2 mmol)
in absolute ethanol (20 mL) containing a catalytic amount of tri-
ethylamine (1 mL), phenyl isothiocyanate 11 (0.27 g, 2 mmol) was
added. The reaction mixture, in each case, was heated under reflux
for 8–10 h until all the reactants had disappeared as indicated by
TLC. The reaction mixture poured over an ice/water mixture and
neutralized with dilute hydrochloric acid. The solid product that
formed, in each case, was filtered off, dried and crystallized from
the appropriate solvent.
C41H47N3O (597.831): C, 82.37; H, 7.92; N, 7.03; found: C, 82.18;
H, 8.07; N, 6.85.
2.1.3.16. 17-[4ꢀ,5ꢀ-Dihydro-5ꢀ-methyl-4ꢀ-(p-chlorophenylamino)-
3ꢀ-phenyl-1ꢀH-pyrazol-5ꢀ-yl]androst-4-ene-3-one
(28a). Brown
2.1.4.2. 20-Hydroxy-20-[1ꢀ,3ꢀ-diphenyl-4ꢀ-hydroxy-2ꢀ-thioxo-1ꢀ,3ꢀ-
imidazol-5ꢀ-yl]pregn-4-ene-3-one (13a). Brown crystals from
absolute ethanol, yield 0.95 g (82%), mp 175–176 ◦C, IR (KBr,
cm−1): ꢀ = 3542 (2OH), 3028 (CH-aromatic), 2978–2870 (CH-
aliphatic), 1708 (C-3, C O), 1592 (C C), 1197 (C S). 1H NMR
(CDCl3, ppm): ı = 0.98 (s, 3H, CH3-19), 1.07 (s, 3H, CH3-18), 1.37
(s, 3H, CH3), 4.82, 5.03 (2s,2H, 2OH, D2O-exchangable), 5.80 (s,
1H, C4-H), 6.82–7.35 (m, 10H, 2C6H5), M.S. (EI): m/z (%): 582 (M+,
34), 557 (M+−CH3, 32), 271 (C19H27O, 60), 77 (100). Calc. for
crystals from methanol, yield 0.78 g (80%), mp 190–210 ◦C, IR (KBr,
cm−1): ꢀ = 3427–3365 (2NH), 3028 (CH-aromatic), 2969–2875
(CH-aliphatic), 1713 (C-3, C O), 1658 (C N), 1625 (C C). 1H NMR
(DMSO-d6, ppm): ı = 1.02 (s, 3H, CH3-19), 1.17 (s, 3H, CH3-18), 1.30
(s, 3H, CH3), 5.82 (s, 1H, C4-H), 6.37 (dd, 2H-aromatic, JHH = 8 Hz),
7.06 (dd, 2H-aromatic, JHH = 8 Hz), 7.32–7.61 (m, 5H, C6H5), 8.36,
8.97 (2brs, 2H, 2NH, D2O-exchangable). M.S. (EI): m/z (%): 556 (M+,
70), 271 (C19H27O, 43), 126 (100), 111 (34). Calc. for C35H42ClN3O
(556.181): C, 75.58; H, 7.61; N, 7.56; found: C, 75.34; H, 7.40; N,
7.75.
C
36H42N2O3S (582.795): C, 74.19; H, 7.26; N, 4.81; S, 5.50; found:
C, 74.40; H, 7.45; N, 4.52; S, 5.22%.

R.M. Mohareb et al. / Steroids 76 (2011) 1190–1203
1195
2.1.4.3. 20-Hydroxy-20-[4ꢀ-hydroxy-3ꢀ-phenyl-1ꢀ-(p-chlorophenyl)-
2ꢀ-thioxo-1ꢀ,3ꢀ-imidazol-5ꢀ-yl]pregn-4-ene-3-one
(13b). Yellow
JHH = 8 Hz). M.S. (EI): m/z (%): 610 (M+, 39), 595 (M+−CH3, 60), 271
(C19H27O, 100), 77 (28). Calc. for C38H46N2O3S (610.848): C, 74.72;
H, 7.59; N, 4.59, S, 5.25; found: C, 74.46; H, 7.30; N, 4.75; S, 5.50%.
crystals from absolute ethanol, yield 0.96 g (78%), mp 78–80 ◦C,
IR (KBr, cm−1): ꢀ = 3537 (2OH), 3030 (CH-aromatic), 2982–2878
(CH-aliphatic), 1705 (C-3, C O), 1608 (C C), 1195 (C S). 1H NMR
(CDCl3, ppm): ı = 0.88 (s, 3H, CH3-19), 1.12 (s, 3H, CH3-18), 1.34
(s, 3H, CH3), 4.97, 5.20 (2s, 2H, 2OH, D2O-exchangable), 5.82 (s,
1H, C4-H), 6.47 (dd, 2H, aromatic, JHH = 7 Hz), 6.53–7.08 (m, 5H,
C6H5), 7.12 (dd, 2H-aromatic, JHH = 7 Hz). 13C NMR (CDCl3, ppm):
ı = 36.7 (C-1), 32.9, (C-2), 197.6 (C-3), 126.8 (C-4), 171.0 (C-5), 35.6
(C-6), 32.5 (C-7), 37.7 (C-8), 52.6 (C-9), 38.0 (C-10), 22.4 (C-11),
36.8 (C-12), 38.9 (C-13), 56.7 (C-14), 27.6 (C-15), 23.5 (C-16), 55.8
(C-17), 23.2 (C-18), 21.8 (C-19), 68.2 (C-20), 23.5 (C-21), 91.2,
172.4, 53.3 (C-imidazole), 134.4, 129.3, 126.4, 124.3, 132.0, 130.1,
129.5, 127.2 (C-phenyl). M.S. (EI): m/z (%): 618 (M++1, 47), 602
(M+−CH3, 37), 600 (M+−H2O, 100), 271 (C19H27O, 52), 77 (64).
Calc. for C36H41N2O3SCl (617.240): C, 70.05; H, 6.70; N, 4.54; S,
5.20; found: C, 70.30; H, 6.92; N, 4.70; S, 5.37%.
2.1.4.8. 20-Hydroxy-20-[1ꢀ,3ꢀ, 4ꢀ-triphenyl-2ꢀ-thioxo-1ꢀ,3ꢀ-imidazol-
5ꢀ-yl]pregn-4-ene-3-one (31a). Yellow crystals from absolute
ethanol, yield 1.02 g (80%), mp 144–145 ◦C, IR (KBr, cm−1): ꢀ = 3385
(OH), 3042 (CH-aromatic), 2977–2875 (CH-aliphatic), 1697 (C-3,
C
O), 1596 (C C), 1197 (C S). 1H NMR (DMSO-d6, ppm): ı = 0.92 (s,
3H, CH3-19), 1.07 (s, 3H, CH3-18), 1.35 (s, 3H, CH3), 4.87 (s, 1H, OH,
D2O-exchangable), 5.90 (s, 1H, C4-H), 7.05–7.87 (m, 15H, 3C6H5).
13C NMR (DMSO-d6, ppm): ı = 36.2 (C-1), 33.0, (C-2), 198.1 (C-3),
126.6 (C-4), 172.2 (C-5), 35.6 (C-6), 32.4 (C-7), 37.2 (C-8), 53.1 (C-
9), 37.8 (C-10), 22.7 (C-11), 36.4 (C-12), 38.9 (C-13), 57.0 (C-14),
27.6 (C-15), 23.2 (C-16), 55.2 (C-17), 23.9 (C-18), 22.0 (C-19), 68.2
(C-20), 21.8 (C-21), 116.4, 171.5, 113.6 (C-imidazole), 135.6, 134.4,
129.3, 128.5, 126.4, 124.8, 124.0, 132.0, 130.1, 131.2, 129.7, 127.2
(C-phenyl). M.S. (EI): m/z (%): 642 (M+, 45), 627 (M+−CH3, 50), 271
(C19H27O, 65), 77 (100). Calc. for C42H46N2O2S (642.908): C, 78.47;
H, 7.21; N, 4.36; S, 4.99; found: C, 78.66; H, 7.40; N, 4.60, S, 4.73%.
2.1.4.4. 20-Hydroxy-20-[4ꢀ-hydroxy-3ꢀ-phenyl-1ꢀ-(p-
methoxyphenyl)-2ꢀ-thioxo-1ꢀ,3ꢀ-imidazol-5ꢀ-yl]pregn-4-ene-3-one
(13c). Yellow crystals from dioxane, yield 0.84 g (69%), mp
121–122 ◦C, IR (KBr, cm−1): ꢀ = 3540 (2OH), 3025 (CH-aromatic),
2975–2868 (CH-aliphatic), 1710 (C-3, C O), 1587 (C C), 1192
(C S). 1H NMR (DMSO-d6, ppm): ı = 1.03 (s, 3H, CH3-19), 1.18 (s,
3H, CH3-18), 1.33 (s, 3H, CH3), 3.82 (s, 3H, OCH3), 4.92, 5.06 (2s, 2H,
2OH, D2O-exchangable), 5.87 (s, 1H, C4-H), 6.48 (dd, 2H-aromatic,
JHH = 8 Hz), 6.82–7.13 (m, 5H, C6H5), 7.22 (dd, 2H-aromatic,
JHH = 8 Hz). M.S. (EI): m/z (%): 612 (M+, 39), 597 (M+−CH3, 60), 271
(C19H27O, 100), 77 (28). Calc. for C37H44N2O4S (612.821): C, 72.52;
H, 7.24; N, 4.57, S, 5.23; found: C, 72.30; H, 7.49; N, 4.75; S, 5.50%.
2.1.4.9. 20-Hydroxy-20-[3ꢀ,4ꢀ-diphenyl-1ꢀ-(p-chlorophenyl)-2ꢀ-
thioxo-1ꢀ,3ꢀ-imidazol-5ꢀ-yl]pregn-4-ene-3-one
(31b). Orange
powder from DMF, yield 1.12 g (83%), mp 178–180 ◦C, IR (KBr,
cm−1): ꢀ = 3367 (OH), 3040 (CH-aromatic), 2980–2856 (CH-
aliphatic), 1698 (C-3, C O), 1596 (C C), 1190 (C S). 1H NMR
(DMSO-d6, ppm): ı = 0.87 (s, 3H, CH3-19), 1.12 (s, 3H, CH3-18), 1.34
(s, 3H, CH3), 4.97, 5.20 (2s, 2H, 2OH, D2O-exchangable), 5.82 (s,
1H, C4-H), 7.01–7.58 (m, 14H, aromatic-H). M.S. (EI): m/z (%): 679
(M++2, 58), 662 (M+−CH3, 37), 659 (M+−H2O, 100), 271 (C19H27O,
42), 77 (60). Calc. for C42H47N2O2SCl (677.353): C, 74.48; H, 6.70;
N, 4.14; S, 4.73; found: C, 74.30; H, 6.89; N, 4.36; S, 5.98%.
2.1.4.5. 20-Hydroxy-20-[1ꢀ,3ꢀ-diphenyl-4ꢀ-methyl-2ꢀ-thioxo-1ꢀ,3ꢀ-
imidazol-5ꢀ-yl]pregn-4-ene-3-one (22a). Pale yellow crystals from
absolute ethanol, yield 0.83 g (72%), mp 156–158 ◦C, IR (KBr, cm−1):
ꢀ = 3413 (OH), 3044 (CH-aromatic), 2977–2862 (CH-aliphatic),
1696 (C-3, C O), 1596 (C C), 1198 (C S). 1H NMR (DMSO. d6,
ppm): ı = 0.92 (s, 3H, CH3-19), 1.07 (s, 3H, CH3-18), 1.37 (s, 6H,
2CH3), 4.87 (s, 1H, OH, D2O-exchangable), 5.90 (s, 1H, C4-H),
7.13–7.62 (m, 10H, 2C6H5). M.S. (EI): m/z (%): 581 (M++1, 45), 549
(M+−CH3, 32), 271 (C19H27O, 56), 77 (100). Calc. For C37H44N2O2S
(580.823): C, 76.51; H, 7.64; N, 4.82; S, 5.52; found: C, 76.72; H,
7.45; N, 4.65, S, 5.30%.
2.1.4.10. 20-Hydroxy-20-[3ꢀ,4ꢀ-diphenyl-1ꢀ-(p-methoxyphenyl)-2ꢀ-
thioxo-1ꢀ,3ꢀ-imidazol-5ꢀ-yl]pregn-4-ene-3-one (31c). Pale brown
crystals from methanol, yield 1.14 g (85%), mp 225–227 ◦C, IR
(KBr, cm−1): ꢀ = 3383 (OH), 3035 (CH-aromatic), 2960–2885
(CH-aliphatic), 1705 (C-3, C O), 1581 (C C), 1184 (C S). 1H NMR
(DMSO-d6, ppm): ı = 1.03 (s, 3H, CH3-19), 1.18 (s, 3H, CH3-18),
1.33 (s, 3H, CH3), 3.82 (s, 3H, OCH3), 4.97, (2s, 2H, 2OH, D2O-
exchangable), 5.87 (s, 1H, C4-H), 6.93–7.53 (m, 14H, aromatic-H).
M.S. (EI): m/z (%): 672 (M+, 32), 657 (M+−CH3, 40), 271 (C19H27O,
100), 77 (36). Calc. for C43H50N2O3S (672.917): C, 76.75; H, 7.19;
N, 4.16, S, 4.77; found: C, 76.96; H, 7.35; N, 4.32; S, 4.53.
2.1.4.6. 20-Hydroxy-20-[4ꢀ-methyl-3ꢀ-phenyl-1ꢀ-(p-chlorophenyl)-
2ꢀ-thioxo-1ꢀ,3ꢀ-imidazol-5ꢀ-yl]pregn-4-ene-3-one (22b). Pale brown
powder from methanol, yield 1.01 g (82%), mp 180–183 ◦C, IR
(KBr, cm−1): ꢀ = 3427 (OH), 3037 (CH-aromatic), 2982–2878
(CH-aliphatic), 1709 (C-3, C O), 1606 (C C), 1192 (C S). 1H NMR
(CDCl3, ppm): ı = 0.88 (s, 3H, CH3-19), 1.15 (s, 3H, CH3-18), 1.39 (s,
6H, 2CH3), 5.07, 5.27 (2s, 2H, 2OH, D2O-exchangable), 5.76 (s, 1H,
C4-H), 6.57 (dd, 2H-aromatic, JHH = 7 Hz), 6.89–7.14 (m, 5H, C6H5)
7.27 (dd, 2H-aromatic, JHH = 7 Hz). M.S. (EI): m/z (%): 616 (M++1,
28), 600 (M+−CH3, 37), 597 (M+−H2O, 100), 271 (C19H27O, 52), 77
(64). Calc. for C37H43N2O2SCl (615.267): C, 72.23; H, 7.04; N, 4.55;
S, 5.21; found: C, 72.45; H, 7.23; N, 4.39; S, 5.50%.
2.2. Pharmacological assay
2.2.1. Animals
Sprague-Dawley strain rats weighing 120–130 g or Swiss albino
mice 20–25 g body weight were used throughout the experiments,
supplied by the Animal House Colony of the National Research
Centre, Cairo, Egypt and acclimated for one week in a specific
pathogen-free (SPF) barrier area where temperature is 25 1 ◦C
and humidity is 55%. Animals were controlled constantly with 12 h
light/dark cycle at the National Research Centre animal facility
breeding colony. Animals were individually housed with ad libi-
tum access to standard laboratory diet and tap water. All animal
procedures were performed after approval from the Ethics Com-
mittee of the National Research Centre and in accordance with the
recommendations for the proper care and use of laboratory animals
(NIH publication No. 85-23, revised 1985).
2.1.4.7. 20-Hydroxy-20-[4ꢀ-methyl-3ꢀ-phenyl-1ꢀ-(p-
methoxyphenyl)-2ꢀ-thioxo-1ꢀ,3ꢀ-imidazol-5ꢀ-yl]pregn-4-ene-3-one
(22c). Yellow crystals from absolute ethanol, yield 0.95 g (78%),
mp 132–134 ◦C, IR (KBr, cm−1): ꢀ = 3480 (OH), 3028 (CH-aromatic),
2945–2882 (CH-aliphatic), 1699 (C-3, C O), 1601 (C C), 1186
(C S). 1H NMR (DMSO-d6, ppm): ı = 1.03 (s, 3H, CH3-19), 1.09 (s,
3H, CH3-18), 1.35 (s, 3H, CH3), 3.78 (s, 3H, OCH3), 4.97, (2s, 2H,
2OH, D2O-exchangable), 5.90 (s, 1H, C4-H), 6.48 (dd, 2H-aromatic,
JHH = 8 Hz), 6.99–7.06 (m, 5H, C6H5), 7.22 (dd, 2H-aromatic,
2.2.2. Tests of Inflammation: carrageenan-induced paw oedema
assay
Paw oedema was induced by sub-plantar injection of 100 ␮L of
1% sterile carrageenan lambda in saline into the right hind paw of

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R.M. Mohareb et al. / Steroids 76 (2011) 1190–1203
rats [20]. Contralateral paw received an equal volume of saline. Paw
volume was determined immediately before carrageenan injec-
tion and at selected times thereafter using a plethysmometer (Ugo
Basile, Milan, Italy). The oedema component of inflammation was
quantified by measuring the paw volume (mL) at zero time (before
carrageenan injection) and at 1, 2, 3 and 4 h after carrageenan
injection and comparing it with the pre-injection value for each
animal. Oedema was expressed as a percentage of change from
control (pre-drug, zero time) values. The effect of systemic admin-
istration of compounds 8b, 19b, 24, and 31a (25 or 50 mg/kg, s.c.,
0.2 mL, n = 6/group) given 30 min before induction of inflamma-
tion by subplantar carrageenan was studied. All tested compounds
administrated in saline (0.9% NaCl) with one drop of Tween 80
as a vehicle in all pharmacological studies. The control group of
carrageenan-treated rats received an equal volume of saline 30 min
before subplantar carrageenan injection (n = 6 each). Another group
administered indomethacin (18 mg/kg, s.c.) served as control pos-
itive.
that with a probability of P < 0.001 were considered to be highly
significant.
3. Results and discussion
3.1. Chemistry
Oxiranes and azoles represent molecular frameworks those
serve as platform for developing pharmaceutical agents for var-
ious applications. Many derivatives of these rings proved as
anti-inflammatory and/or anti-nociceptive agents [23–26]. Intro-
duction of the reactive oxirane in steroids results in dramatic
changes in their biological activity [27]. We have attempted
a straightforward synthesis of 17-(oxiran-2ꢀ-yl)androst-4-ene-3-
one derivatives, progesterone (1) reacted with equimolar amount
of ethyl chloroacetate 2 in potassium tert-butoxide via Darzens
condensation to afford the ethyl 17-(oxiran-2ꢀ-yl)androst-4-ene-
3ꢀ-carboxylate derivative 3 in 72% yield (Scheme 1). In this
conversion, the selective epoxidation took place only at C-20
carbonyl moiety in progesterone. No product containing oxirane
functionality at C-3 position was found. The mass spectrum (EIMS)
of compound 3 revealed the presence of molecular ion peak at
m/z = 400 (29%) and the IR spectrum showed two carbonyl groups
stretching at ꢀ = 1708 cm−1 (C-3) and ꢀ = 1742 cm−1 (acetate, C O).
Also the 1H NMR spectrum of compound 3 revealed, in addition
to the expected signals of pregnene moiety, the presence of sin-
glet signal at ı = 3.45 ppm (1H) for the C-3 proton of oxirane ring
and showed also triplet at ı = 1.37 (3H) and quartet at ı = 4.25 (2H)
which are characteristic for the ethyl ester group. To generalize
such a methodology, the previous reaction was carried out by using
the ␣-haloketones, chloroacetone 14 or ␣-bromoacetophenone
23 under the same experimental conditions, to afford the cor-
responding 17-(oxiran-2ꢀ-yl)androst-4-ene-3-one derivative 15 in
78% yield (Scheme 2) and 24 in 74% yield, respectively (Scheme 3).
2.2.3. Tests of nociception
2.2.3.1. Hot plate assay. The hot-plate test was performed using
an electronically controlled hot plate (Ugo Basile, Italy) heated to
53 ◦C ( 0.1 ◦C). Each mouse was placed unrestrained on hot plate
for the baseline measurement just prior to saline or drug adminis-
tration. Different groups of mice (n = 6/group) were given one of
compounds 8b, 19b, 24 or 31a (25 or 50 mg/kg; 0.2 mL, orally),
tramadol (20 mg/kg, 0.2 mL, orally) (control +ve) or saline (control
−ve). Measurements were then taken 60 min after drug adminis-
tration. The experimenter was blind to doses. Latency to lick a hind
paw or jump out of the apparatus was recorded for the control and
drug-treated groups. The cut-off time was 30 s.
2.2.3.2. Acetic acid induced writhing. Separate groups of 6 mice
each were administered saline, compound 8b, 19b, 24 or 31a (25
or 50 mg/kg; 0.5 mL, orally) or indomethacin (18 mg/kg, 0.5 mL,
orally). After 60 min, mice received an i.p. injection of 0.6%
acetic acid (0.2 mL) [21]. The number of writhes (constrictions of
abdomen, twisting of trunk and extension of hind legs) during
30 min observation period following acetic acid injection was com-
pared with the control group and drug-treated groups.
The reaction of compound
3 with either aniline 4a, p-
chloroaniline 4b or p-methoxyaniline 4c in refluxing absolute
ethanol containing a catalytic amount of triethylamine afforded
the corresponding ethyl androst-4-en-arylaminobutanoate deriva-
tives 5a, 5b, and 5c, respectively (Scheme 1). Similarly under
the same experimental condition, compounds 15 and 24 reacted
with either aniline 4a, p-chloroaniline 4b, or p-methoxyaniline
4c to afford the corresponding 17-(3ꢀ-arylaminobutyl)androst-4-
ene-3-one derivatives 16a–c (Scheme 2) and 25a–c, respectively
(Scheme 3).
The azole moiety often shows some special biological activ-
ity when it is introduced to some biologically active compounds
[28,29]. The basicity and hydrophilicity of an azole in theory might
alter the biological function of a steroid [30]. The reactivity of
compounds 5a–c towards the reaction with some nucleophilic
reagents was studied in the aim of forming new pyrazolyl steroids.
Thus, compounds 5a–c reacted with either hydrazine hydrate 6a
or phenyl hydrazine 6b in ethanolic triethylamine to give the
corresponding 17-(pyrazol-5ꢀ-yl)androst-4-ene-3-one derivatives
8a,b, 9a,b and 10a,b, respectively. The reaction takes place via
the non-isolable intermediates 7a–f which undergo intermolecu-
lar cyclization to afford the isolable products 8a,b, 9a,b and 10a,b
(Scheme 1). Similarly, compounds 16a–c and 25a–c reacted with
either hydrazine hydrate 6a or phenyl hydrazine 6b to give the
corresponding 17-(pyrazol-5ꢀ-yl)androst-4-ene-3-one derivatives
18a,b, 19a,b, 20a,b (Scheme 2) and 27a,b, 28a,b, 29a,b (Scheme 3),
respectively. Elucidation of proposed structures of the latter prod-
ucts was based on their correct elemental analysis and compatible
IR, 1H NMR, 13C NMR and mass spectral data (cf. Section 2).
The reactivity of compounds 5a–c towards the reaction with
isothiocyanates was studied in the aim of forming new imidazolyl
2.2.4. Gastric ulcerogenic study
Gastric mucosal damage was evoked in rats by the administra-
tion of indomethacin (20 mg/kg, 0.2 mL, s.c.). The effect of either
compound 8b, 19b, 24 or 31a (50 mg/kg; 0.5 mL, orally) admin-
istered at time of indomethacin injection was studied. Rats were
killed 24 h after indomethacin administration. In other experi-
ments, the effect of tested compounds (50 mg/kg; 0.5 mL, s.c.) on
gastric damage caused by ethanol (96%) was evaluated. Rats were
fasted for 18 h, but allowed water ad libitum. They were adminis-
tered either saline (control) or tested compounds 30 min prior to
ethanol (96%, 1 mL, p.o.). Rats were killed 1 h after ethanol admin-
istration, stomachs excised, opened along the greater curvature,
rinsed with saline, extended on a plastic board and examined for
mucosal lesions. The number and severity of mucosal lesions were
noted and lesions were scaled as described by Mózsik et al. [22].
2.3. Statistical analysis
Data were expressed as mean SE. Data were analyzed by one-
way analysis of variance, followed by a Tukey’s multiple range tests
for post hoc comparison of group means. When there were only two
groups a two-tailed Student’s t-test was used. For all tests, effects
with a probability of P < 0.05 were considered to be significant and

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1197
O
H3C
O
COOEt
K/ t.butyl alc
ClCH2COOEt
+
2
O
3
1
H3C
OH
NHAr
NH2
R
COOEt
EtOH / Et3N
4a, R=H
b, R=Cl
c, R=OMe
5a, Ar=Ph
b, Ar=C6H4- p-Cl
c, Ar=C6H4- p-OMe
R1
N
H3C
OH
N
NHAr
CONHNHR1
H3C
OH
- H2O
NHAr
EtOH / Et3N
5 (a-c)
RNHNH2
6a, R=H
b, R=Ph
+
7a, Ar= Ph., R1=H
8a, Ar=Ph., R1=H
8b, Ar=Ph., R1=Ph
b, Ar= Ph., R1=Ph
c, Ar= C6H4- p-Cl, R1=H
9a, Ar=C6H4- p-Cl, R1=H
9b, Ar=C6H4- p-Cl, R1=Ph
10a, Ar=C6H4- p-OMe, R1=H
d, Ar= C6H4- p-Cl, R1=Ph
e, Ar= C6H4- p-OMe, R1=H
f, Ar= C6H4- p-OMe, R1=Ph
10b, Ar=C6H4- p-OMe, R1=Ph
Ar
Ar
N
S
S
H3C
OH
N
H3C
OH
C
NHPh
N
COOEt
Ph
HO
EtOH, Et3N
-EtOH
5 (a-c)
PhNCS
11
+
13a, Ar= Ph
12a, Ar= Ph
b, Ar= C6H4- p-Cl
b, Ar= C6H4- p-Cl
c, Ar= C6H4- p-OMe
c, Ar= C6H4- p-OMe
Scheme 1.
also absorption peaks at ꢀ = 1197, 1195 and 1192 cm−1 for C
S
steroids. Thus, compounds 5a–c reacted with phenylisothiocyanate
11 in refluxing absolute ethanol containing a catalytic amount
of triethylamine to afford the corresponding 20-(imidazol-5ꢀ-
yl)pregn-4-en-3-one derivatives 13a–c. Formation of the latter
products is explained in terms of the intermediate formation of
12a–c (Scheme 1). The IR spectra of compounds 13a–c revealed
broad absorption peaks equivalent to the OH groups and showed
groups, respectively. The mass spectra of compounds 13a–c
revealed the presence of molecular ion peaks at m/z = 582 (34%),
618 (M+−1, 47%), and 612 (39%), respectively. Also, the 13C
NMR spectrum of compound 13b showed the presence of the
C
S carbon at ı = 172.4. Similarly, the reaction of compounds
16a–c and 25a–c with phenylisothiocyanate 11 under the same

1198
R.M. Mohareb et al. / Steroids 76 (2011) 1190–1203
H3C
O
COCH3
K / t.butyl alc
ClCH2COCH3
+
1
14
15
H3C
OH
NHAr
NH2
R
COCH3
EtOH / Et3N
4a, R=H
b, R=Cl
c, R=OMe
16a, Ar=Ph
b, Ar=C6H4- p-Cl
R1
N
c, Ar=C6H4- p-OMe
H3C
OH
N
NHAr
C NNHR
CH3
H3C
CH3
- H2O
NHAr
EtOH / Et3N
16 (a-c) RNHNH2
+
6a, R=H
b, R=Ph
17a, Ar= Ph., R1=H
b, Ar= Ph., R1=Ph
c, Ar= C6H4- p-Cl, R1=H
d, Ar= C6H4- p-Cl, R1=Ph
e, Ar= C6H4- p-OMe, R1=H
f, Ar= C6H4- p-OMe, R1=Ph
18a, Ar=Ph., R1=H
18b, Ar=Ph., R1=Ph
19a, Ar=C6H4- p-Cl, R1=H
19b, Ar=C6H4- p-Cl, R1=Ph
20a, Ar=C6H4- p-OMe, R1=H
20b, Ar=C6H4- p-OMe, R1=Ph
Ar
Ar
N
S
S
H3C
OH
N
H3C
OH
C
NHPh
N
COCH3
Ph
H3C
EtOH, Et3N
- H2O
16 (a-c)
PhNCS
11
+
22a, Ar= Ph
21a, Ar= Ph
b, Ar= C6H4- p-Cl
b, Ar= C6H4- p-Cl
c, Ar= C6H4- p-OMe
c, Ar= C6H4- p-OMe
Scheme 2.
experimental conditions described before, afforded the corre-
sponding 20-(imidazol-5ꢀ-yl)pregn-4-en-3-one derivatives 22a–c
(Scheme 2) and 31a–c (Scheme 3), respectively. The structures of
all new compounds were assigned by both correct elemental and
spectral data (cf. Section 2).
It is interesting to note that most of the compounds synthe-
sized in this work can exist in either Z or E structures. However,
according to the concept of push–pull alkenes reviewed by Sand-
strom [31], conjugated systems with unsaturated heterocyclic
rings with conjugated alkyl chains, exemplify typical push–pull
compounds, which can exist in Z/E equilibrium depending on
the temperature and the nature of solvent. One may say that
all the newly synthesized compounds in this work are typi-
cal push–pull compounds existing in Z/E equilibrium. Isolation
and identification of these isomers is beyond the scope of this
study.

R.M. Mohareb et al. / Steroids 76 (2011) 1190–1203
1199
H3C
O
COPh
K / t.butyl alc
PhCOCH Br
+
1
2
23
24
H3C
OH
NHAr
COPh
NH2
R
EtOH / Et N
3
4a, R=H
b, R=Cl
c, R=OMe
25a, Ar=Ph
b, Ar=C H4- p-Cl
c, Ar=C6H4- p-OMe
R1
N
6
H3C
OH
N
NHAr
H3C
Ph
C
NNHR
- H2O
NHAr
EtOH / EtN
3
Ph
25 (a-c) RNHNH
+
2
6a, R=H
b, R=Ph
26a, Ar= Ph., R=H
27a, Ar=Ph., R=H
1
1
b, Ar= Ph., R=Ph
27b, Ar=Ph., R=Ph
c, Ar= C H4-1p-Cl, R =H
28a, Ar=C H4-1p-Cl, R =H
6
1
6
1
d, Ar= C H4- p-Cl, R =Ph
28b, Ar=C H4- p-Cl, R =Ph
e, Ar= C6H4- p-OMe, R =H
1
29a, Ar=C6H4- p-OMe1, R =H
f, Ar= C6H4- p-OMe, R1=Ph
6
1
29b, Ar=C H4- p-OMe, R =Ph
6
1
6
1
Ar
Ar
N
S
S
H3C
OH
N
H3C
OH
C
NHPh
N
COPh
Ph
Ph
EtOH, Et N
3
- H2O
25 (a-c)
PhNCS
11
+
31a, Ar= Ph
30a, Ar= Ph
b, Ar= C H4- p-Cl
c, Ar= C6H4- p-OMe
b, Ar= C H4- p-Cl
c, Ar= C6H4- p-OMe
6
6
Scheme 3.
3.2. Pharmacology
observed carefully for all adverse symptoms. There were not any
symptoms of toxicity during the study at all employed doses.
The most structurally promising of the novel steroidal het-
erocyclic derivatives, compounds 8b, 19b, 24 and 31a were
investigated individually as anti-inflammatory, anti-nociceptive
and anti-ulcerogenic agents. During the study period, animals are
3.2.1. Anti-inflammatory assay
The effects of systemic injection of the tested compounds on
oedema formation were studied using a carrageenan induced

1200
R.M. Mohareb et al. / Steroids 76 (2011) 1190–1203
Table 1
The anti-inflammatory effect of the tested compounds on carrageenan induced paw oedema.
Group
Basal
0.28 0.006
1 h
2 h
3 h
4 h
Control
(Saline)
0.53 0.02 (88.3 7.0)
0.49 0.02 (60.8 6.1)
0.458 0.02(53.7 4.1)*
0.576 0.03 (78.9 5.2)
0.51 0.04 (54.3 4.4)*
0.55 0.003 (80.1 2.1)
0.445 0.02(48.8 3.5)*
0.59 0.009 (96.9 4.5)
0.53 0.04 (78.4 5.5)
0.6 0.018 (107.8 7.6)
0.51 0.02 (66.9 6.3)*
0.496 0.01 (66.8 5.6)*
0.546 0.012(68.9 6.4)*
0.553 0.026(65.1 4.8)*
0.535 0.011(73.7 4.5)*
0.483 0.02 (61.3 5.9)*
0.546 0.011(82.5 5.4)*
0.453 0.013(50.6 3.5)*
0.59 0.013 (112.2 7.3)
0.475 0.017(55.7 5.1)*
0.456 0.016(53.3 4.8)*
0.537 0.012(65.8 6.4)*
0.575 0.019(73.2 5.4)*
0.498 0.01 (62.0 5.4)*
0.457 0.016(52.0 4.9)*
0.52 0.011 (74.1 5.0)*
0.45 0.013 (53.4 4.2)*
0.60 0.013 (113.3 8.4)
0.475 0.013(55.7 4.4)*
0.415 0.019(39.3 2.3)*
0.513 0.009(58.5 5.4)*
0.516 0.004(55.6 2.9)*
0.486 0.012(58.2 5.8)*
0.43 0.011 (43.9 3.9)
0.49 0.012 (63.4 4.8)*
0.44 0.013 (50.6 4.7)*
8b (25 mg/kg)
8b (50 mg/kg)
19b (25 mg/kg)
19b (50 mg/kg)
24 (25 mg/kg)
24 (50 mg/Kg)
31a (25 mg/kg)
31a (50 mg/kg)
0.305 0.004
0.298 0.004
0.325 0.008
0.335 0.014
0.308 0.004
0.301 0.01
0.3 0.004
0.29 0.003
Indomethacin (18 mg/kg)
0.288 0.004
0.48 0.02 (67.4 5.6)*
0.47 0.01 (64.6 4.10)*
0.435 0.019(51.4 4.3)*
0.415 0.015(44.4 2.9)*
Results are expressed as percentage change for control (pre-drug) values. Data are expressed as mean S.E., n = 6 rats/group. Asterisks indicate significant change from control
value. The s.c. administration of all tested compounds inhibited the carrageenan induced paw oedema (two-way ANOVA; treatment effect: F13,280 = 30; P < 0.001; time effect:
F3,280 = 23.8; P < 0.001, time × drug effect: F39,280 = 3; P < 0.001). The values in parenthesis indicate the percentage (%) of increase in paw volume from basal (zero time) values.
paw inflammation. Each compound was injected sucutaneously
(s.c.), in two doses (25 or 50 mg/kg), 30 min before sub-plantar
carrageenan. All tested compounds decreased paw oedema in
dose-dependent manner compared to the control group (pre-drug)
(Table 1). Indomethacin was given at 18 mg/kg, s.c., 30 min before
carrageenan as positive control. Data are expressed as mean S.E.,
n = 6 per group. The values in parenthesis in Table 1 indicate the per-
centage (%) of increase in paw volume (oedema) from basal (zero
time) values.
3.2.1.2. Effect of compounds 19b and 31a. Compounds 19b and 31a
at the dose of 25 mg/kg significantly inhibited the paw oedema
response compared with the control group at 2, 3 and 4 h post-
carrageenan, the percent of oedema inhibition was (−36, −41.4,
−48.4%) and (−23.5, −33.9, −44%), respectively. The higher dose
(50 mg/kg) of compound 31a significantly inhibited oedema by
(−53.1, −52.4, −55.3) and that of compound 19b significantly inhib-
ited oedema by (−39.6, −34.8, −51%) at 2, 3 and 4 h time points,
respectively (Fig. 2).
3.2.1.1. Effect of compounds 8b and 24. The oedema response was
significantly reduced by the administration of compounds 8b or
24 at dose of 25 or 50 mg/kg. The two compounds at the dose of
25 mg/kg markedly and significantly inhibited the paw oedema
response compared with the control group at 2, 3 and 4 h post-
carrageenan where the percent of inhibition of oedema was (−37.9,
−50.4, −50.8%) for compound 8b and (−31.6, −44.7, −48.6%) for
compound 24. The higher dose (50 mg/kg) of compound 8b or
24 significantly inhibited oedema, at 1, 2, 3 and 4 h time points,
by (−38, −52.2, −65.3%) and (−43.1, −53.6, −61.3%), respectively
(Fig. 1).
3.2.2. Tests of anti-nociceptive studies
3.2.2.1. Effect of tested compounds on thermal pain. The hot plate
latency was significantly increased denoting analgesic effect after
1 h of the administration of either compound 8b or 19b at both
administered doses (Table 2, Fig. 3). While, the administration of
compound 24 or 31a significantly increased the hot plate latency
only at the dose of 50 mg/kg compared to the saline-treated group.
The anti-nociceptive effect was most marked with compound
19b with a maximal increase in hot-plate latency by 72.1% after
drug administration at 50 mg/kg (Table 2, Fig. 3). Compound 19b
Carrageenan control
+ 8b (25 mg/kg)
+ 8b (50 mg/kg)
+ 24 (25 mg/kg)
+ 24 (50 mg/kg)
+ Indomethacin (18 mg/kg)
Carrageenan control
+ 31a (25 mg/kg)
+ 31a (50 mg/kg)
140
+ 19b (25 mg/kg)
140
120
100
80
+ 19b (50 mg/kg)
+ Indomethacin (18 mg/kg)
120
100
80
60
40
20
0
*
*
*
*
*
*
*
*
*
*
*
*
60
*
40
20
0
0
1
2
3
4
5
0
1
2
3
4
5
Time (h)
Time (h)
Fig. 1. Effect of tested compounds 8b or 24 on the carrageenan paw oedema forma-
tion. Compound 8b or 24 was given (25 or 50 mg/kg, i.p.) 30 min prior to carrageenan
injection and rats were evaluated for paw oedema at 1, 2, 3 and 4 h post-carrageenan.
The results are expressed as a percentage change from control (pre-drug) values,
each point represents the mean S.E. of six rats per group. Asterisks indicate signif-
icant change from the control group at the corresponding time point.
Fig. 2. Effect of tested compounds 31a or 19b on the carrageenan paw oedema
formation. Compound 31a or 19b were given (25 or 50 mg/kg, i.p.) 30 min prior to
carrageenan injection and rats were evaluated for paw oedema at 1, 2, 3 and 4 h post-
carrageenan. The results are expressed as a percentage change from control (pre-
drug) values, each point represents the mean S.E. of six rats per group. Asterisks
indicate significant change from the control group at the corresponding time point.

R.M. Mohareb et al. / Steroids 76 (2011) 1190–1203
1201
Table 2
Percentage increase in hot plate latency in mice treated with tested compounds in
comparison with tramadol.
Drug
0 time (basal)
1 h
% Change
Saline
13.2 0.6
14.38 1.3
15.0 0.7
13.46 1.2
13.76 1.0
13.56 1.1
14.24 0.66
14.64 0.76
14.28 0.81
12.86 1.1
14.65 1.1
18.4 1.2
19.1 1.0
18.62 1.3
23.68 1.6
15.98 0.9
17.29 1.5
16.94 1.3
17.52 1.2
20.3 1.6
8b (25 mg/kg)
8b (50 mg/kg)
19b (25 mg/kg)
19b (50 mg/kg)
24 (25 mg/kg)
24 (50 mg/kg)
31a (25 mg/kg)
31a (50 mg/kg)
Tramadol (20 mg/kg)
27.96*
27.33*
38.3**
72.1**
17.8
21.4*
15.7
22.7*
57.9**
Data are expressed as mean S.E., n = 6 per group.
*
P < 0.05 vs corresponding basal values.
P < 0.01 vs corresponding basal values.
**
Fig. 4. Effect of tested compounds on acetic acid-induced writhing. Test com-
pounds were administered at 25 or 50 mg/kg and the number of abdominal writhes
induced by i.p. injection of acetic acid in mice was determined over 30 min period
(mean S.E. of 6 mice/group). The percent decrease in the number of writhes from
the saline control group is represented above the respective group bar. Asterisks
indicate significant change from the saline control group (ANOVA and Duncun’s
multiple comparison tests).
compared to the saline-treated control group. The analgesic activ-
ity of either dose of compound 8b, 24 or 31a as well as the high dose
of 19b was significantly higher than that for indomethacin. In addi-
tion, the 50 mg/kg dose of compound 24 or 31a was significantly
more potent as analgesic vs that of compound 19b.
3.2.3. Tests of gastric ulcerogenic studies
In order to evaluate the potential anti-ulcerogenic properties of
the tested compounds, we examined the effect of the highest dose
on the development of gastric mucosal lesions caused by ethanol
or the non-steroidal anti-inflammatory drug indomethacin.
Gastric mucosal lesions caused in the rats by the administration
of 96% ethanol were inhibited by all tested compounds adminis-
tered at (50 mg/kg) dose in the study (Table 4, Fig. 5). Compounds
19b, 24, 31a had significant effect vs the control and compound 8b.
Also, compounds 19b and 31a had significant effect vs compound
24. Gastric mucosal lesions caused in the rats by the administration
of indomethacin were inhibited by all tested compounds adminis-
tered at 50 mg/kg in the study (Table 5, Fig. 6).
Fig. 3. Reaction time on the hot-plate in seconds after the administration of tested
compounds at doses of 25 or 50 mg/kg. Shown are basal (pre-drug: first column)
and 60 min (post-drug: second column) values. The percentage change from basal
(pre-drug) values is shown (n = 6/group). Asterisks indicate significant change from
the saline control group at the respective time point (ANOVA and Duncan’s multiple
comparison tests).
at 50 mg/kg was significantly more prolonged than tramadol at
20 mg/kg (57.9%).
3.2.2.2. Effect of tested compounds on acetic acid-induced writhing.
All tested compounds significantly reduced the number of abdom-
inal writhes induced by i.p. administration of dilute acetic acid
in mice (Table 3, Fig. 4). The degree of inhibition of the writhing
response by these compounds ranged from −55.0% to −99.5% as
3.3. Conclusion
In this study we have described a straightforward and efficient
synthesis of novel steroid derivatives containing fused oxiran or
azole nucleus in addition to the pharmacophoric features of the
steroid moiety. We investigated also the pharmaceutical impor-
tance of incorporating heterocyclic moiety to the steroid nucleus
Table 3
Effect of tested compounds on the number of writhes in the acetic acid test in mice.
Group
Number of abdominal
constrictions/30 min
% Inhibition vs
control
Table 4
Effect of the tested compounds on gastric mucosal injury caused by 96% ethanol in
rats.
Saline
80.0 5.3
7.2 0.5*
7.0 0.7*
31.0 2.3*
17.5 1.8*
0.4 0.24*
1.0 0.36*
9.3 0.8*
1.83 0.7*
38 2.8*
8b (25 mg/kg)
8b (50 mg/kg)
19b (25 mg/kg)
19b (50 mg/kg)
24 (25 mg/kg)
24 (50 mg/kg)
31a (25 mg/kg)
31a (50 mg/kg)
IND (18 mg/kg)
91.0%
91.3%
61.3%
78.2%
99.5%
98.8%
88.3%
97.8%
52.5%
Group
Number of lesions/rat
Severity of lesions/rat
Ethanol control
Ethanol + 8b
Ethanol + 19b
Ethanol + 24
Ethanol + 31a
20.6 2.1
16.4 2.0NS
1.2 1.2*
9.0 1.6 *
1.4 0.7*
63.8 4.0
58.6 6.8NS
1.2 1.2*
30.2 2.9*
2.8 1.2*
Statistical comparison of the difference between the ethanol control group and other
treated groups is indicated by asterisks; *P < 0.05, NS = not significant vs control
values.
Data are expressed as means and S.E.M. (n = 6/group). IND: indomethacin.
*
P < 0.05 vs control values.

1202
R.M. Mohareb et al. / Steroids 76 (2011) 1190–1203
to form new effective hybrid molecules. The novel synthesized
derivatives 8b, 19b, 24, and 31a showed anti-inflammatory, anti-
nociception and anti-ulcerogenic activities with various intensities.
Oedema was significantly reduced by both doses (25 or 50 mg/kg)
of tested compounds at 3 and 4 h post-carrageenan. Compound 19b
was most effective in alleviating thermal pain. The analgesic activ-
ity of either dose of compound 8b, 24 or 31a as well as the high
dose of 19b was significantly higher than that for indomethacin.
Gastric mucosal lesions caused in the rats by the administration of
96% ethanol or indomethacin were inhibited by any of the tested
compounds administered at 50 mg/kg. These results provide a
unique opportunity to develop new anti-inflammatory drugs which
lack the ulcerogenic liabilities associated with currently marketed
drugs. Finally, the encouraging results displayed by these com-
pounds are of interest to initiate further studies of the mechanism
of action and toxicity profile of the promising tested compounds
before application in phase 1 of clinical study in the hope of finding
new potent prescriptions
Fig. 5. Effect of test compounds administrated at 50 mg/kg, on the number and
severity of gastric mucosal lesions caused by s.c. injection of EtOH (96%) in rats.
The percentage decrease in the number or severity of gastric lesions from the EtOH
(96%) control group is represented above the respective group bar. Asterisks indicate
significant change from the corresponding EtOH (96%) control group (ANOVA and
Duncan’s multiple comparison tests).
Acknowledgements
The authors express their thanks to Prof. Dr. Selim F. Estefan,
Hormones Dept., National Research Center, for critically reviewing
the manuscript. The authors acknowledge the financial support of
the National Research Center, Egypt (grant no. E-8040505-2008-
2010).
Table 5
Effect of the tested compounds on gastric mucosal injury caused by indomethacin
in rats.
References
Group
Number of lesions/rat
Severity of lesions/rat
[1] Kozuch PL, Hanauer SB. Treatment of inflammatory bowel disease: a review of
medical therapy. World J Gastroenterol 2008;14(3):354–77.
[2] Sondhi SM, Singhal N, Johar M, Reddy BSN, Lown JW. Heterocyclic compounds
as inflammation inhibitors. Curr Med Chem 2002;9:1045–74.
[3] Gureje O, Von Knoff M, Simon GE, Gater R. Persistent pain and well-being: a
World Health Organization study in primary care. JAMA 1998;280:147–51.
[4] Karthikeyan K, Veenus Seelan T, Lalitha KG, Perumal PT. Synthesis and antinoci-
ceptive activity of pyrazolyl isoxazolines and pyrazolyl isoxazoles. Bioorg Med
Chem Lett 2009;19:3370–3.
IND (control)
IND + 8b
IND + 19b
IND + 24
3.6 0.6
0.4 0.2*
0.0 0.0*
0.2 0.2*
0.0 0.0*
5.3 0.8
0.4 0.2*
0.0 0.0*
1.2 0.5*
0.0 0.0*
IND + 31a
Statistical comparison of the difference between the indomethacin (IND) control
group and other treated groups is indicated by asterisks; *P < 0.05 vs control values.
[5] Wallace JL, Soldato PD, Cirino G. Development of NSAIDs with reduced gas-
trointestinal and renal toxicity. Exp Opin Invest Drugs 1995;4:613–9.
[6] Sadeghi S, Bain CJ, Pandeya N, Webb PM, Green AC, Whiteman DC. Aspirin, non-
steroidal anti-inflammatory drugs, and the risks of cancers of the esophagus.
Cancer Epidemiol Biomarkers Prev 2008;17:1169–78.
[7] Lipworth BJ. Systemic adverse effects of inhaled corticosteroid therapy: a sys-
tematic review and meta-analysis. Arch Intern Med 1999;159:941–55.
[8] Phillips GH. Mechanism of topical corticoid activity. Edinburgh, U.K.: Living-
stone; 1981. p. 1.
[9] Criscuolo D, Fraioli F, Bonifacio V. Effects of a new glucocorticoid, deflazacort,
on pituitary–adrenal function in man: a comparison with prednisone. Int J Clin
Pharmacol Ther Toxicol 1980;18:37–41.
[10] Hoyte RM, Zhang J, Lerum R, Oluyemi A, Persaud P, O’Connor C, et al. Synthesis of
halogen-substituted pyridyl and pyrimidyl derivatives of [3,2-c]pyrazolo corti-
costeroids: strategies for the development of glucocorticoid receptor mediated
imaging agents. J Med Chem 2002;45:5397–405.
[11] Wüst F, Carlson KE, Katzenellenbogen JA. Synthesis of novel arylpyrazolo cor-
ticosteroids as potential ligands for imaging brain glucocorticoid receptors.
Steroids 2003;68:177–91.
[12] Elmegeed GA, Wardakhan WW, Baiuomy AR. Synthesis of thiazolyl and thieno
cholestane derivatives. A novel class of potent anti-inflammatory steroids.
Pharmazie 2005;60:328–33.
[13] Elmegeed GA, Baiuomy AR, Abdel-Salam OME. Evaluation of the anti-
inflammatory and anti-nociceptive activities of novel synthesized melatonin
analogues. Eur J Med Chem 2007;42:1285–92.
[14] Wardakhan WW, Elmegeed GA, Abdel-Salam OME. New approaches for the
synthesis of fused benzothiophene derivatives: new class of antidepressant,
sedative or analgesic agents. Acta Pharm 2007;58:1–14.
[15] IUPAC. Joint Commission on Biochemical Nomenclature (JCBN). Nomenclature
of steroids. Pure Appl Chem 1989;61:1783–822.
[16] IUPAC. Joint Commission on Biochemical Nomenclature (JCBN). The nomencla-
ture of steroids, recommendations 1989. Eur J Biochem 1989;186:429–58.
[17] Gacs-Baitz E, Minuti L, Taticchi A. Synthesis and complete 1H and 13C NMR
analysis of some 4-androsten-3-one derivatives. Synop 1996;27:324–5.
[18] Frenkel M, Marsh KN. TRC data series: spectral data for steroids. Texas, TX:
College Station, Thermodynamics Research Center; 1994, 386 pp.
[19] Johnson WS, Schneider WP. Ethyl indole-2-carboxylate. Org Synth
1963;4:132–6.
Fig. 6. Effect of tested compounds administrated at 50 mg/kg on the number and
severity of gastric mucosal lesions caused by s.c. injection of indomethacin in rats.
The percentage decrease in the number or severity of gastric lesions from the
indomethacin (IND) control group is represented above the respective group bar.
Asterisks indicate significant change from the corresponding IND control group
(ANOVA and Duncan’s multiple comparison tests).

R.M. Mohareb et al. / Steroids 76 (2011) 1190–1203
1203
[20] Winter CA, Risley EA, Nuss GW. Carrageenan-induced edema in hind paw
of the rat as an assay for antiinflammatory drugs. Proc Soc Exp Biol Med
1962;111:544–52.
[21] Koster R, Anderson M, De Beer EJ. Acetic acid for analgesic screening. Fed Proc
1959;18:412–3.
[22] Mózsik GY, Móron F, Jávor T. Cellular mechanisms of the development of gastric
mucosal damage and of gastrocytoprotection induced by prostacyclin in rats.
A pharmacological study. Prostaglandins Leukot Med 1982;9:71–84.
[23] Rostom SAF, El-Ashnawy IM, Abd El Razik HA, Badr MH, Ashour HMA. Design
and synthesis of some thiazolyl and thiadiazolyl derivatives of antipyrine as
potential non-acidic anti-inflammatory, analgesic and antimicrobial agents.
Bioorg Med Chem 2009;17:882–95.
[24] Sauzem PD, Sant’anna GS, Machado P, Duarte MM, Ferreira J, Mello CF, et al.
Effect of 5-trifluoromethyl-4,5-dihydro-1H-pyrazoles on chronic inflamma-
tory pain model in rats. Eur J Pharmacol 2009;616:91–100.
[26] Amer H, Mereiter K, Stanetty C, Hofinger A, Czollner L, Beseda I, et al.
Synthesis and crystal structures of ring A modified glycyrrhetinic acid deriva-
tives derived from 2,3-oxirane and 2,3-thiirane intermediates. Tetrahedron
2010;66(26):4390–402.
[27] Gella IM, Gladkova AI, Zolotukhina VA, Karpenko NA. Local antiandrogenic
activity of steroid 3-spirooxiranes. Pharm Chem J 1995;29(11):753–6.
[28] Tanitame A, Oyamada Y, Ofuji K, Fujimoto M, Suzuki K, Ueda T, et al. Synthesis
and antibacterial activity of novel and potent DNA gyrase inhibitors with azole
ring. Bioorg Med Chem 2004;12:5515–24.
[29] Hackett JC, Kim YW, Su B, Brueggemeier RW. Synthesis and characterization of
azole isoflavone inhibitors of aromatase. Bioorg Med Chem 2005;13:4063–70.
[30] Wong FF, Chen C, Chen T, Huang J, Fang H, Yeh M. Synthesis of 3alpha-
hydroxy-21-(1ꢀ-imidazolyl)-3beta-methoxyl-methyl-5alpha-pregnan-20-one
via lithium imidazole with 17alpha-acetylbromopregnanone. Steroids
2006;71:77–82.
[25] Tseng C, Lin C, Shih P, Tsao L, Wangd J, Cheng C, et al.
Fluro[3ꢀ,2ꢀ:3,4]naphtho[1,2-d]imidazole derivatives as potential inhibitors of
inflammatory factors in sepsis. Bioorg Med Chem 2009;17:6773–9.
[31] Sandstrom JT. Measurement and calculation of [sup.13]C and [sup.15]N
NMR chemical-shift tensors of
1983;14:83–181.
a push–pull ethylene. Top Stereochem