Synthesis and antiandrogenic activity of some new 3-substituted androstano[17,16-c]-5′-aryl-pyrazoline and their derivatives

Article abstract of DOI:10.1016/j.bmc.2005.08.024

A series of androstano[17,16-c]pyrazolines and their oxidized derivatives (2-14) were synthesized using 3β-hydroxyandrostan-17-one as a starting material (1a, b). The synthesized compounds were evaluated for their antiandrogenic activity compared to that

Full text of DOI:10.1016/j.bmc.2005.08.024

Bioorganic & Medicinal Chemistry 14 (2006) 373–384
Synthesis and antiandrogenic activity of some new 3-substituted
androstano[17,16-c]-50-aryl-pyrazoline and their derivatives
Abd El-Galil E. Amr,a,* Nehad A. Abdel-Latifb and Mohamed M. Abdallac
aApplied Organic Chemistry Department, National Research Centre, Dokki, Cairo, Egypt
bNatural Compounds Chemistry Department, National Research Centre, Dokki, Cairo, Egypt
cResearch Unit, Egyptian Pharmacist Co., Cairo, Egypt
Received 7 May 2005; revised 7 August 2005; accepted 9 August 2005
Available online 22 September 2005
Abstract—A series of androstano[17,16-c]pyrazolines and their oxidized derivatives (2–14) were synthesized using 3b-hydroxyandro-
stan-17-one as a starting material (1a, b). The synthesized compounds were evaluated for their antiandrogenic activity compared to
that of Cyproteroneꢀ as positive control. Some of the compounds exhibited better antiandrogenic activity than the reference drug.
The detailed synthesis, spectroscopic data, ED50, and toxicity (LD50 and LD90) of the synthesized compounds were reported.
ꢁ 2005 Elsevier Ltd. All rights reserved.
1. Introduction
2. Results and discussion
In a previous work, we found that certain substituted
steroidal derivatives showed androgenic, anabolic, and
anti-inflammatory activities.1 Some new heterocyclic
compounds containing nitrogen atom have been synthe-
sized and used as antiparkinsonian,2 antitumor,3–5
antimicrobial,6–8 and anti-inflammatory9,10 activities.
Pyrazolines present an interesting group of compounds,
many of which possess widespread pharmacological
properties such as analgesic, antipyretic, and antirheu-
matic activities.11,12 These derivatives are also well
known for their pronounced anti-inflammatory proper-
ties13,14 and are used as potent antidiabetic agents.15,16
In addition, the pharmacological and antitumor activi-
ties of many compounds containing pyrazoline rings
have been reviewed.17–19 Recently, the heterocyclic
nitrogen derivatives exhibited a general ionophoric
potency for divalent cations20 and used a novel thiocy-
anate-selective membrane sensor.21 In view of these re-
ports and in continuation of our previous work in
heterocyclic chemistry, we have synthesized some new
compounds containing pyrazoline ring fused with
steroidal structure for the evaluation of antiandrogenic
activity compared to Cyproteroneꢀ as standard drug.
2.1. Chemistry
The arylmethylenes 1 and 2 are in the E-form (thermo-
dynamically favored, less sterically hindered, and with
a more favored formation of the intermediate resonance
stabilized anion). Based on the assumption of Barton
et al.22–24 the preparation of E-isomer results from Aldol
condensation of steroidal and terpinoidal ketones with
aromatic aldehydes.
Other evidence for the formation of E-isomer came from
the pmr-trace where the downfield shift of the arylidene
proton (7.75–7.81 ppm) indicated an E-isomer (7.61 for
Z-isomer).
Also, the starting materials were the (8b,9a,14a) 3b-
hydroxyandrost-5-ene-3-one (i.e., the absolute configura-
tion is 5a,14a). In other words, A/B ring fusion is a and
C/D ring fusion is a. This configuration was retained
in all reaction process, where the aromatic protons of
the steroidal nucleus resonate at their normal values.25
In the condensation of hydrazine with the arylidene sys-
tem after removal of one molecule of water and formation
of the 17-hydrazone, the NH2 attacks the double bond
from a-face leading to the formation of 16R, and the
phenyl ring rotates backward forming 50-S center and
all the asymmetric centers of the starting material are
retained (no change in chemical shift in the pmr-trace).
Keywords: Pyrazolines; Moffat oxidation, Antiandrogenic activity;
Cyperoterone.
*
Corresponding author. Tel.: +20 24 528 69 4; fax: +2 02 33 71 9 01;
e-mail: aamr1963@yahoo.com
0968-0896/$ - see front matter ꢁ 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2005.08.024

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Abd El-Galil E. Amr et al. / Bioorg. Med. Chem. 14 (2006) 373–384
The evidence for the 16R center is that when C–C car-
bon bond between C-16 and C-50 is b, the C18 carbon
atom appears deshielded at more than 0.7 ppm (cf. at
0.65 ppm for 16a-substituent).26
ring at room temperature with trifluoroacetic anhydride
to give the corresponding 3b-trifluoroacetate-16-arylm-
ethylene-androstan-17-one (2a,b), which was condensed
with hydrazine hydrate in refluxing glacial acetic acid or
propionic acid to afford the 3b-trifluoroacetate-androst-
ano-N-substituted pyrazolines (3a,b) and (4a,b), respec-
tively (Scheme 1).
Also, here 16aH, 50bH (due to backward rotation of the
phenyl group to a less sterically hindered cage) is eluci-
dated via determination of Jvic for the interaction be-
tween C-16 and C-50 protons. The dihedral angle
measurement from the model and the constant of the
original Karplus equations (equations 3–5) was 120ꢂ.
Compounds 3 and 4 were hydrolyzed by refluxing in
alcoholic potassium hydroxide 5% to yield 3b-hydroxy-
androstanopyrazoline derivatives 5a,b, which can also
be obtained directly by condensation of arylmethylene
derivatives 1a,b with hydrazine hydrate in refluxing eth-
anol (Scheme 1).
J16aHꢀ5 bH ꢁ 1:2 CPS (cf. ꢁ1 CPS) if 16bH.27
0
Other evidence for a single isomer came from GC–MS
that showed only a single peak compound in the chro-
matogram and HPLC that showed also high purity for
each compound (not less than 98%).
Oxidation of 3b-hydroxyandrostanopyrazoline deriva-
tives 5a,b with Moffat oxidizing agent in the presence
of dicyclohexylcarbodiimide at room temperature gave
the corresponding 3-oxo-androstenopyrazoline deriva-
tives 6a,b, which were acetylated with acetylchloride to
afford the corresponding 3-oxo-androsteno-N-acetylpy-
Arylmethylenes of 3b-hydroxyandrostan-17-one deriva-
tives (1a,b) were synthesized according to a reported
procedure.1 Compounds (1a,b) were protected by stir-
O
O
Ar
(CF3CO)2O
H
Ar
H
H
H
O
H
H
HO
H
(1a,b)
F3C
O
H
(2a,b)
O
CH3
N
N
H
Ar
H
H
NH2NH2
O
H
H
Propionic acid
F3C
O
H
(4a,b)
NH2NH2
Acetic acid
O
H
N
N
H
CH3
Method A
5% KOH
Ar
H
O
H
H
F3C
N
O
(3a,b)
H
NH
H
H
Method B
Method A
Ar
H
NH2NH2
5% KOH
EtOH
H
H
HO
H
(5a,b)
a, Ar = C H ; b, Ar = C H OCH (p)
6 5
6
4
3
Scheme 1.

Abd El-Galil E. Amr et al. / Bioorg. Med. Chem. 14 (2006) 373–384
375
razoline derivatives 7a,b. Also, oxidation of compounds
2.2. Biological assay
7a,b by dichlorodicyanoquinolinone (DDQ) in the pres-
ence of bubbled anhydrous hydrogen chloride gas in
dioxane afforded 3-oxo-1,4-diene-androsteno-N-acetyl-
pyrazolines (8a,b), which were hydrolyzed in refluxing
5% alcoholic potassium hydroxide to yield 3-oxo-1,4-di-
ene androstanopyrazoline derivatives 9a,b (Scheme 2).
2.2.1. Antiandrogenic activity. All synthesized com-
pounds were tested for their antiandrogenic activities
in vivo. Additionally, ED50, LD50, and LD90 were deter-
mined. All the tested compounds showed antiandrogen-
ic activity. The logarithmic concept was used to
calculate the relative potency according to complex con-
cept that seemed to be unreliable for the referee so we
recalculate it as he suggests (Table 1). All compounds
showed antiandrogenic activities, but less than that of
Cyproteroneꢀ.
On the other hand, condensation of compounds (2a,b)
with substituted hydrazines, namely, methylhydrazine
or phenylhydrazine, in refluxing glacial acetic acid gave
the corresponding 3b-trifluoroacetateandrostano-N-
substituted pyrazoline derivatives (10a–d), which were
hydrolyzed by 5% alcoholic potassium hydroxide to af-
ford 3b-hydroxyandrostano-N-substituted pyrazoline
derivatives (11a–d) (Scheme 3). Oxidation of com-
pounds 11a–b using the Moffat method gave 3-oxo-
2.2.2. Acute toxicity. The LD50 and LD90 (rats) were
determined by injecting different increasing doses and
calculating the dose that kills 50% and 90%, respectively,
of the animals.
androstano-N-substituted
pyrazoline
derivatives
(12a–d), which were oxidized with dichlorodicyanoquin-
olinone (DDQ) to afford the corresponding 3-oxo-1,
4-diene androstano-N-substituted pyrazolines 13a–d,
while condensation of compounds 12a–d with
ethylene triphenylphosphorane by heating at 60 ꢂC in
dimethylsulfoxide for 12 h afforded 3-ethylene androst-
3. Experimental
3.1. Synthesis
Melting points were uncorrected and were recorded with
an Electrothermal IA 9000 Digital Melting Point Appa-
ratus. Microanalytical data were obtained from the
ano-N-substituted
(Scheme 3).
pyrazoline
derivatives
14a–d
N
NH
H
N
NH
H
H
M
offat oxidation
Ar
H
Ar
H
H
H
H
H
H
HO
O
(5a,b)
H
H
(6a,b)
CH3COCl
O
O
N
N
H
CH3
N
N
H
CH3
H
H
DDQ
O
Ar
Ar
H
H
H
H
H
H
O
(7a,b)
(8a,b)
H
N
NH
H
Ar
H
H
5% KOH
H
H
O
(9a,b)
a, Ar = C H ; b, Ar = C H OCH (p)
6 4
6
5
3
Scheme 2.

376
Abd El-Galil E. Amr et al. / Bioorg. Med. Chem. 14 (2006) 373–384
R
N
N
H
H
Ar
R
NHNH2
H
(2a,b)
glacial acetic acid
O
H
H
F3C
O
H
(10a-d)
5% KOH
R
R
N
N
H
N
N
H
H
H
Moffot oxidation
Ar
Ar
H
H
H
H
H
H
H
O
HO
H
(12a-d)
(11a-d)
DDQ
Q3P=CHCH3
R
N
N
H
R
H
N
N
H
H
Ar
H
Ar
H
H
H
H
H
H
O
H3C
(14a-d)
(13a-d)
Scheme 3.
Microanalytical Unit, Cairo University, Egypt, and
their results were found to be in an agreement with pro-
posed structures. The IR spectra (KBr) were recorded
on a FT IR-8201 PC Spectrophotometer (Shimadzu).
The 1H NMR spectra were measured with Jeol
270 MHz in DMSO-d6 or CDCl3 and the chemical shifts
were recorded in d ppm relative to TMS. The mass spec-
tra (EI) were run at 70 eV with a Finnigan SSQ GC/MS
spectrometer. The reactions were followed by TLC (sil-
ica gel, aluminum sheets 60 F254, Merck). HPLC Shi-
madzu (pump SCL 10 AVP and detector SPD 10
AVP), Japan, column package L3, 4.6 nm · 25 cm, flow
rate 1 ml/min, UV detector 254 nm. The starting materi-
als 1a,b were prepared according to the published
procedures.1
For standard preparation transfer, ꢁ12 mg of the tested
agents is transferred into a glass-stoppered 50 ml conical
flask and 25 ml of internal standard solution is added,
sonicated for 5 min, and 1 ml of this solution and 3 ml
of mobile phase are combined (see Table 2).
3.1.1. Synthesis of (8b,9a,14a)-3b-trifluoroacet-16(E)-ar-
yl-methylene-5a-androstan-17-one (2a,b). A mixture of
the arylmethylene derivatives (1a,b)1 (1 mmol) and tri-
fluoroacetic anhydride (5 ml) was kept overnight at
room temperature. The reaction mixture was evaporated
under reduced pressure up to dryness and the obtained
residue was solidified with 1 N sodium bicarbonate
(10 ml). The solid formed was collected by filtration,
washed with water, dried, and crystallized from metha-
nol to give 3-trifluoroacetate derivatives (2a,b).
Mobile phase is butyl chloride, water saturated butyl
chloride, THF, methanol, and glacial acetic acid
[95:95:14:7:6]. Internal standard is a solution of methyl
parben in mobile phase having a concentration of
0.04 mg/ml.
3.1.1.1.
(8b,9a,14a)-3b-Trifluoroacet-16(E)-phenyl-
methylene-5a-androstan-17-one (2a). Yield 78%, mp:
25
D
231 ꢂC (MeOH); ½aꢂ +91 (c 1, MeOH); IR (KBr): 1738,
1660 cmꢀ1. 1H NMR (CDCl3): d 0.85 (s, 3H, CH3), 0.95

Abd El-Galil E. Amr et al. / Bioorg. Med. Chem. 14 (2006) 373–384
377
Table 1. Evaluation of the ED50, LD50, LD90, and antiandrogenic activity of the newly synthesized compounds
Compound No.
ED50 mg/kg
LD50 mg/kg
LD90 mg/kg
Relative antiandrogenic potency to Cyproteroneꢀ (100%)
2a
2b
6.55 0.048
6.18 0.049
6.14 0.047
6.32 0.051
6.25 0.044
6.12 0.048
6.18 0.041
6.21 0.042
5.13 0.039
5.40 0.038
5.30 0.039
5.20 0.037
4.18 0.039
4.12 0.038
3.91 0.039
4.00 0.038
7.15 0.051
7.14 0.051
6.44 0.044
6.13 0.041
6.80 0.041
5.00 0.041
5.10 0.041
4.41 0.044
4.12 0.041
3.91 0.031
4.10 0.031
3.20 0.029
3.10 0.022
3.56 0.023
3.18 0.031
3.14 0.032
2.16 0.031
2.15 0.030
2.30 0.031
2.10 0.032
1.70 0.031
630 0.019
655 0.018
710 0.018
753 0.019
673 0.019
663 0.017
708 0.019
725 0.019
813 0.019
800 0.019
730 0.019
741 0.020
980 0.020
1012 0.031
1109 0.031
983 0.033
688 0.019
791 0.019
890 0.018
830 0.018
810 0.017
940 0.019
813 0.019
866 0.019
851 0.017
861 0.016
638 0.016
813 0.017
647 0.018
618 0.015
613 0.016
823 0.016
941 0.017
1012 0.018
1210 0.018
1216 0.017
1518 0.016
1190 0.011
1188 0.011
1214 0.011
1413 0.011
1111 0.011
1088 0.012
1213 0.013
1340 0.013
1460 0.017
1416 0.012
1318 0.012
1322 0.018
1680 0.019
1730 0.017
1814 0.016
1704 0.016
1210 0.016
1314 0.016
1560 0.014
1340 0.016
1550 0.017
1630 0.016
1510 0.016
1630 0.016
1740 0.016
1810 0.017
1430 0.015
1510 0.013
1280 0.012
1380 0.013
1310 0.014
1460 0.015
1805 0.016
1811 0.013
1930 0.011
2031 0.014
2200 0.012
25.95
27.50
27.68
26.89
27.20
27.77
27.50
27.37
33.13
30.14
32.07
32.69
40.66
41.62
43.47
42.50
23.77
23.80
26.39
27.73
27.50
34.00
33.33
38.54
41.26
43.47
41.46
53.12
54.83
47.75
53.45
54.14
78.70
79.06
73.91
73.85
80.95
3a
3b
4a
4b
5a
5b
6a
6b
7a
7b
8a
8b
9a
9b
10a
10b
10c
10d
11a
11b
11c
11d
12a
12b
12c
12d
13a
13b
13c
13d
14a
14b
14c
14d
Cyproteroneꢀ
Error limits are SEMÕS.
(s, 3H, CH3), 0.98–1.00 (m, 1H, CH), 1.28–1.30 (m, 4H,
2CH2), 1.40–1.60 (m, 6H, 3CH2), 1.64–1.90 (m, 4H,
2CH2), 2.00 (m, 1H, CH), 2.24–2.40 (m, 2H, CH2), 2.45
(m, 1H, CH), 2.55 (m, 1H, 3a-CH), 3.20 (m, 1H, 5a-
CH), 7.35–7.45 (m, 5H, Ar-H), 7.81 (s, 1H, C@CH). MS
(EI): m/z 474 (15%) [M+]. Anal. Calcd for C28H33F3O3:
C, 70.86; H, 7.01. Found: C, 70.78; H, 7.00.
ylene derivatives (2a,b) (4 mmol) and hydrazine hydrate
(0.8 ml, 16 mmol) in glacial acetic acid or propionic acid
(15 ml) was refluxed for ꢁ7 h. The reaction mixture was
poured onto ice water and neutralized with sodium
bicarbonate. The formed precipitate was collected by
filtration, washed with water, dried, and crystallized
from the proper solvent to give the corresponding
N-substituted pyrazoline derivatives (3a,b) and (4a,b),
respectively.
3.1.1.2. (8b,9a,14a)-3b-Trifluoroacet-16(E)-(p-meth-
oxyphenyl)-methylene-5a-androstan-17-one (2b). Yield
25
62%, mp: 216 ꢂC (MeOH); ½aꢂ +96 (c 1, MeOH); IR
3.1.2.1. (16R,50S)(8b,9a,14a)-3b-Trifluoroacetate-5a-
D
1
(KBr): 1735, 1650 cmꢀ1. H NMR (CDCl3): d 0.81 (s,
3H, CH3), 0.90 (s, 3H, CH3), 0.95–1.05 (m, 1H, CH),
1.25–1.28 (m, 4H, 2CH2), 1.36–1.58 (m, 6H, 3CH2),
1.62–1.88 (m, 4H, 2CH2), 1.95 (m, 1H, CH), 2.20–2.35
(m, 2H, CH2), 2.48 (m, 1H, CH), 2.54 (m, 1H, 3a-
CH), 3.10 (m, 1H, 5a-CH), 3.65 (s, 3H, OCH3), 7.20–
7.40 (m, 4H, Ar-H), 7.75 (s, 1H, C@CH). MS (EI): m/
z 519 (25%) [M+]. Anal. Calcd for C29H35F3O4: C,
69.03; H 6.99. Found: C, 68.98; H, 6.94.
androstano[17,16-c]-10H-50-phenyl-N-acetylpyrazoline
25
(3a). Yield 65%, mp: 167 ꢂC (MeOH); ½aꢂ +85 (c 1,
D
1
MeOH); IR (KBr): 1745, 1665, 1600 cmꢀ1. H NMR
(CDCl3): d 0.78 (s, 3H, CH3), 0.85 (s, 3H, CH3),
0.90–0.98 (m, 1H, CH), 1.25–1.30 (m, 4H, 2CH2),
1.38–1.56 (m, 6H, 3CH2), 1.60–1.92 (m, 4H, 2CH2),
2.00 (m, 1H, CH), 2.08 (s, 3H, COCH3), 2.20–2.35
(m, 2H, CH2), 2.40 (m, 1H, CH), 2.50 (m, 1H, pyraz-
oline-H), 2.55 (m, 1H, 3a-CH), 3.00 (m, 1H, 5a-CH),
3.35 (d, 1H, pyrazoline-H), 7.30–7.60 (m, 5H, Ar-H).
MS (EI): m/z 530 (10%) [M+]. Anal. Calcd for
C30H37F3N2O3: C, 67.90; H, 7.03; N, 5.28. Found: C,
67.86; H, 7.00; N, 5.25.
3.1.2. Synthesis of (16R,50S) (8b,9a,14a)-3b-trifluoro-
acet-10H,5a-androstano-[17,16-c]-50-aryl-N-substituted-
pyrazoline (3a,b) and (4a,b). A mixture of the arylmeth-

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Abd El-Galil E. Amr et al. / Bioorg. Med. Chem. 14 (2006) 373–384
Table 2. HPLC of the synthesized compounds
2.36 (m, 2H, CH2), 2.40 (m, 1H, pyrazoline-H), 2.48
(m, 1H, CH), 2.55 (m, 1H, 3a-CH), 3.10 (m, 1H, 5a-
CH), 3.50 (d, 1H, pyrazoline-H), 7.30–7.50 (m, 5H,
Ar-H). MS (EI): m/z 544 (20%) [M+]. Anal. Calcd for
C31H39F3N2O3, C, 68.36; H, 7.22; N, 5.14. Found: C,
68.32; H, 7.18; N, 5.10.
Compound No.
Relative retention time
Purity
1a
1b
1.0
1.1
99.81
99.13
98.16
98.74
98.28
98.74
98.16
98.14
99.88
99.95
99.19
98.16
99.18
99.58
99.71
99.03
98.06
99.14
99.17
98.90
99.88
99.61
99.71
99.63
98.91
98.81
98.56
98.66
99.15
99.07
99.48
99.32
99.38
99.16
99.56
99.65
99.16
99.11
2a
2b
1.28
1.2
3a
3b
1.24
1.7
3.1.2.4. (16R,50S)(8b,9a,14a)-3b-Trifluoroacetate-5a-
androstano[17,16-c]-10H-50-(p-methoxyphenyl)-N-propo-
4a
4b
1.78
1.18
1.13
1.85
1.40
1.17
1.16
1.51
1.56
1.48
1.62
1.37
1.31
1.22
1.73
1.63
1.30
1.13
1.47
1.08
1.11
1.29
1.66
1.77
1.63
1.46
1.41
1.12
1.15
1.33
1.22
1.66
nyl-pyrazoline (4b). Yield 55%, mp: 274 ꢂC (MeOH);
25
5a
5b
½aꢂ +151 (c 1, MeOH); IR (KBr): 1735, 1670,
D
1
1630 cmꢀ1. H NMR (DMSO-d6): d 0.82 (s, 3H, CH3),
0.85 (s, 3H, CH3), 0.90–0.95 (m, 1H, CH), 1.24–1.30
(m, 4H, 2CH2), 1.45 (t, 3H, CH3), 1.50–1.55 (m, 6H,
3CH2), 1.60–1.85 (m, 4H, 2CH2), 1.95 (m, 1H, CH),
2.05 (q, 2H, CH2), 2.18–2.35 (m, 2H, CH2), 2.35 (m,
1H, pyrazoline-H), 2.45 (m, 1H, CH), 2.50 (m, 1H, 3a-
CH), 3.05 (m, 1H, 5a-CH), 3.40 (d, 1H, pyrazoline-H),
3.70 (s, 3H, OCH3), 7.30–7.45 (m, 4H, Ar-H). MS
(EI): m/z 574 (18%) [M+]. Anal. Calcd for
C32H41F3N2O4: C, 66.88; H, 7.19, N, 4.88. Found: C,
66.82; H, 7.15; N, 4.82.
6a
6b
7a
7b
8a
8b
9a
9b
10a
10b
10c
10d
11a
11b
11c
11d
12a
12b
12c
12d
13a
13b
13c
13d
14a
14b
14c
14d
3.1.3. Synthesis of (16R,50S)(8b, 9a,14a)-3b-Hydroxy-
(5a,b).
5a-androstano[17,16-c]-10H-50-aryl-pyrazoline
Method A: A solution of the appropriate compounds
(3a,b) or (4a,b) (4 mmol) in alcoholic potassium hydrox-
ide (5%, 15 ml) was refluxed for 2–4 h. The reaction mix-
ture was acidified with hydrochloric acid, and the
obtained solid was filtered off, washed with water, dried,
and crystallized from the proper solvent to give com-
pounds (5a,b).
Method B: A mixture of the arylidenes (1a,b) (4 mmol)
and hydrazine hydrate (16 mmol) in ethanol (25 ml)
was refluxed for ꢁ5 h. The solvent was evaporated un-
der reduced pressure to dryness, and the residue was
solidified with water, filtered off, washed with water,
dried, and crystallized from methanol to give com-
pounds (5a,b).
3.1.2.2. (16R,50S)(8b,9a,14a)-3b-Trifluoroacetate-5a-
androstano[17,16-c]-10H-50-(p-methoxyphenyl)-N-acetyl-
3.1.3.1. (16R,50S)(8b,9a,14a)-3b-Hydroxy-5a-androst-
pyrazoline (3b). Yield 60%, mp: 268 ꢂC (EtOH/AcOEt);
ano[17,16-c]-10H-50-phenyl-pyrazoline (5a). Yield 65%,
25
D
25
½aꢂ
+80 (c 1, MeOH); IR (KBr): 1740, 1660,
.
mp: 251 ꢂC (AcOEt); ½aꢂ +65 (c 1, MeOH); IR (KBr):
D
1
1600 cmꢀ1
1H NMR (CDCl3): d 0.8 (s, 3H, CH3),
3435, 3230, 1610 cmꢀ1. H NMR (DMSO-d6): d 0.75
(s, 3H, CH3), 0.90 (s, 3H, CH3), 0.95–1.00 (m, 1H, CH),
1.24–1.28 (m, 4H, 2CH2), 1.40–1.60 (m, 6H, 3CH2),
1.65–1.85 (m, 4H, 2CH2), 1.98 (m, 1H, CH), 2.25
(m, 1H, pyrazoline-H), 2.28–2.45 (m, 2H, CH2), 2.50
(m, 1H, CH), 2.55 (m, 1H, 3a-CH), 3.00 (m, 1H,
5a-CH), 3.45 (d, 1H, pyrazoline-H), 7.30–7.55 (m, 5H,
Ar-H), 10.10 (s, 1H, NH, exchangeable with D2O),
10.55 (s, 1H, OH, exchangeable with D2O). MS (EI): m/
z 392 (20%) [M+]. Anal. Calcd for C26H36N2O, : C,
79.50; H, 9.17; N, 7.14. Found: C, 79.44; H, 9.12; N, 7.10.
0.90 (s, 3H, CH3), 0.95–1.10 (m, 1H, CH), 1.21–1.28
(m, 4H, 2CH2), 1.41–1.60 (m, 6H, 3CH2), 1.65–1.88
(m, 4H, 2CH2), 2.05 (m, 1H, CH), 2.10 (s, 3H, COCH3),
2.18–2.30 (m, 2H, CH2), 2.36 (m, 1H, CH), 2.45 (m, 1H,
pyrazoline-H), 2.58 (m, 1H, 3a-CH), 3.05 (m, 1H, 5a-
CH), 3.25 (d, 1H, pyrazoline-H), 3.65 (s, 3H, OCH3),
7.25–7.50 (m, 4H, Ar-H). MS (EI): m/z 560 (35%)
[M+]. Anal. Calcd for C31H39F3N2O4: C, 66.41; H,
7.01; N, 5.00. Found: C, 66.35; H, 6.98; N, 4.96.
3.1.2.3. (16R,50S)(8b,9a,14a)-3b-Trifluoroacetate-5a-
androstano[17,16-c]-10H-50-phenyl-N-proponylpyrazoline
3.1.3.2. (16R,50S)(8b,9a,14a)-3b-Hydroxy-5a-androst-
25
D
1
(4a). Yield 45%, mp: 261 ꢂC (AcOEt); ½aꢂ +98 (c 1,
ano[17,16-c]-10H-50-(p-methoxy-phenyl)pyrazoline (5b).
25
MeOH); IR (KBr): 1730, 1668, 1630 cmꢀ1. H NMR
(CDCl3): d 0.85 (s, 3H, CH3), 0.92 (s, 3H, CH3), 0.95–
1.00 (m, 1H, CH), 1.22–1.28 (m, 4H, 2CH2), 1.50 (t,
3H, CH3), 1.55–1.60 (m, 6H, 3CH2), 1.65–1.90 (m, 4H,
2CH2), 1.98 (m, 1H, CH), 2.10 (q, 2H, CH2), 2.20–
Yield 70%, mp: 188 ꢂC (MeOH); ½aꢂ +95 (c 1, MeOH);
D
IR (KBr): 3440, 3240, 1620 cmꢀ1. 1H NMR (DMSO-d6):
d 0.80 (s, 3H, CH3), 0.85 (s, 3H, CH3), 0.90–0.95 (m, 1H,
CH), 1.20–1.26 (m, 4H, 2CH2), 1.42–1.62 (m, 6H,
3CH2), 1.60–1.90 (m, 4H, 2CH2), 2.05 (m, 1H, CH),

Abd El-Galil E. Amr et al. / Bioorg. Med. Chem. 14 (2006) 373–384
379
2.30 (m, 1H, pyrazoline-H), 2.30–2.40 (m, 2H, CH2),
2.48 (m, 1H, CH), 2.52 (m, 1H, 3a-CH), 3.10 (m, 1H,
5a-CH), 3.30 (d, 1H, pyrazoline-H), 3.65 (s, 3H,
OCH3), 7.20–7.40 (m, 4H, Ar-H), 9.85 (s, 1H, NH,
exchangeable with D2O), 10.40 (s, 1H, OH, exchange-
able with D2O). MS (EI): m/z 422 (30%) [M+]. Anal.
Calcd for C27H38N2O2: C, 76.73; H, 9.06; N, 6.63.
Found: C, 76.70; H, 8.98; N, 6.58.
bicarbonate, and the obtained solid was filtered off,
washed with water, dried, and crystallized from the
proper solvent to give compounds (7a,b).
3.1.5.1. (16R,50S)(8b,9a,14a)-3-Oxo-5a-androstano[17,16-
c]-10H-50-phenyl-N-acetyl-pyrazoline (7a). Yield 68%,
25
mp: 230 ꢂC (MeOH); ½aꢂ +156 (c 1, MeOH); IR
D
1
(KBr): 1745, 1670, 1640 cmꢀ1. H NMR (DMSO-d6):
d 0.85 (s, 3H, CH3), 1.0 (s, 3H, CH3), 1.05–1.15 (m,
1H, CH), 1.22–1.28 (m, 4H, 2CH2), 1.40–1.61 (m, 6H,
3CH2), 1.64–1.90 (m, 4H, 2CH2), 1.98 (m, 1H, CH),
2.20 (s, 3H, COCH3), 2.25–2.38 (m, 2H, CH2), 2.40
(m, 1H, pyrazoline-H), 2.52 (m, 1H, CH), 3.05 (m,
1H, 5a-CH), 3.45 (d, 1H, pyrazoline-H), 7.20–7.40 (m,
5H, Ar-H). MS (EI): m/z 432 (60%) [M+]. Anal. Calcd
for C28H36N2O2: C, 77.73; H, 8.39; N, 6.48. Found: C,
77.68; H, 8.34; N, 6.40.
3.1.4. Synthesis of (16R,50S)(8b,9a,14a)-3-oxo-5a-and-
rostano[17,16-c]-10H-50aryl-pyrazoline (6a,b). Moffat
method: The appropriate pyrazoline derivatives (5a,b)
(2 mmol) were dissolved in a mixture of [benzene
(3 ml), dimethylsulfoxide (3 ml), pyridine (0.16 ml),
and TFA (0.01 ml)] and then dicyclohexylcarbodiimide
(1.24 g, 6 mmol) was added. The reaction mixture was
kept overnight at room temperature, diethyl ether
(50 ml) was added and then oxalic acid (0.54 g, 6 mmol)
in methanol (5 ml), and after 30 min, water (50 ml) was
added. The obtained dicyclohexylurea was removed by
filtration. The product was extracted from filtrate with
ether and washed with 5% sodium bicarbonate, then
with water. The ethereal solution was dried over anhy-
drous sodium sulfate and evaporated under reduced
pressure. The formed residue was crystallized from eth-
anol to give the corresponding oxo-compounds (6a,b) in
pure form.
3.1.5.2. (16R,50S)(8b,9a,14a)-3-Oxo-5a-androstano[17,16-
(7b).
c]-10H-50-(p-methoxy-phenyl)-N-acetylpyrazoline
25
Yield 60%, mp: 130 ꢂC (MeOH); ½aꢂ +70 (c 1, MeOH);
D
1
IR (KBr): 1740, 1670, 1630 cmꢀ1. H NMR (CDCl3): d
0.80 (s, 3H, CH3), 1.0 (s, 3H, CH3), 1.05–1.10 (m, 1H,
CH), 1.24–1.30 (m, 4H, 2CH2), 1.42–1.60 (m, 6H,
3CH2), 1.62–1.92 (m, 4H, 2CH2), 2.10 (m, 1H, CH),
2.20 (s, 3H, COCH3), 2.30 (m, 1H, pyrazoline-H),
2.32–2.40 (m, 2H, CH2), 2.50 (m, 1H, CH), 3.15 (m,
1H, 5a-CH), 3.35 (d, 1H, pyrazoline-H), 3.65 (s, 3H,
OCH3), 7.20–7.40 (m, 4H, Ar-H). MS (EI): m/z 462
(25%) [M+]. Anal. Calcd for C29H38N2O3: C, 75.28; H,
8.28; N, 6.06. Found: C, 75.18; H, 8.22; N, 6.00.
3.1.4.1. (16R,50S)(8b,9a,14a)-3-Oxo-5a-androstano[17,16-
c]-10H-50-phenylpyrazoline (6a). Yield 55%, mp: 305 ꢂC
25
(MeOH); ½aꢂ +101 (c 1, MeOH); IR (KBr): 3280,
D
1
1740, 1620 cmꢀ1. H NMR (DMSO-d6): d 0.80 (s, 3H,
CH3), 0.85 (s, 3H, CH3), 0.90–0.95 (m, 1H, CH),
1.24–1.28 (m, 4H, 2CH2), 1.40–1.60 (m, 6H, 3CH2),
1.64–1.92 (m, 4H, 2CH2), 2.05 (m, 1H, CH), 2.22–2.40
(m, 2H, CH2), 2.50 (m, 1H, pyrazoline-H), 2.55 (m,
1H, CH), 3.05 (m, 1H, 5a-CH), 3.40 (d, 1H, pyrazo-
line-H), 7.35–7.60 (m, 5H, Ar-H), 10.30 (s, 1H, NH,
exchangeable with D2O). MS (EI): m/z 390 (45%)
[M+]. Anal. Calcd for C26H34N2O: C, 79.95; H, 8.77;
N, 7.17. Found: C, 79.90; H, 8.74; N, 7.14.
3.1.6. Synthesis of (16R,50S)(8b,9a,14a)-3-oxo-androst-
1,4-diene[17,16-c]-10H-50-aryl-N-acetylpyrazoline (8a,b).
Anhydrous hydrogen chloride was bubbled for a few
seconds into a mixture of the compounds (7a,b)
(6 mmol) and dichlorodicyanoquinone (DDQ) (1.8 g)
in dioxane (40 ml). The mixture was kept for 30 min
at room temperature with stirring, the precipitate
hydroquinone was filtered off and the filtrate was
diluted with water, then extracted with ether, the ethe-
real solution was washed with 1% sodium hydroxide,
water, and dried over anhydrous sodium sulfate. The
ether part was evaporated under reduced pressure to
dryness and then crystallized from the proper solvent
to give the corresponding 3-oxo-diene derivatives
(8a,b).
3.1.4.2. (16R,50S)(8b,9a,14a)-3-Oxo-5a-androstano[17,16-
c]-10H-50-(p-methoxy-phenyl)pyrazoline (6b). Yield 80%,
25
mp: 256 ꢂC (MeOH/AcOEt); ½aꢂ +130 (c 1, MeOH);
D
1
IR (KBr): 3245, 1745, 1615 cmꢀ1. H NMR (DMSO-
d6): d 0.75 (s, 3H, CH3), 0.85 (s, 3H, CH3), 0.95–1.00
(m, 1H, CH), 1.25–1.30 (m, 4H, 2CH2), 1.36–1.58 (m,
6H, 3CH2), 1.60–1.88 (m, 4H, 2CH2), 2.10 (m, 1H,
CH), 2.20–2.42 (m, 2H, CH2), 2.25 (m, 1H, pyrazo-
line-H), 2.50 (m, 1H, CH), 3.00 (m, 1H, 5a-CH), 3.35
(d, 1H, pyrazoline-H), 3.70 (s, 3H, OCH3), 7.25–7.50
(m, 4H, Ar-H), 10.35 (s, 1H, NH, exchangeable with
D2O). MS (EI): m/z 420 (25%) [M+]. Anal. Calcd for
C27H36N2O2: C, 77.10; H, 8.63; N, 6.66. Found: C,
76.95; H, 8.58; N, 6.60.
3.1.6.1. (16R,50S)(8b,9a,14a)-3-Oxo-androst-1,4-di-
(8a).
ene[17,16-c]-10H-50-phenyl-N-acetylpyrazoline
Yield 58%, mp: 238 ꢂC (EtOH/pet.ether 40–60 ꢂC);
25
D
½aꢂ
+71 (c 1, MeOH); IR (KBr): 1720, 1665,
.
1630 cmꢀ1 1H NMR (DMSO-d6): d 0.90 (s, 3H,
CH3), 1.05 (s, 3H, CH3), 1.10–1.15 (m, 1H, CH),
1.25–1.30 (m, 4H, 2CH2), 1.40–1.60 (m, 2H, CH2),
1.65–1.70 (m, 2H, CH2), 1.75 (m, 1H, CH), 1.85–
2.00 (m, 2H, CH2), 2.10 (s, 3H, COCH3), 2.25 (m,
1H, pyrazoline-H), 2.30–2.45 (m, 1H, CH), 3.30 (d,
1H, pyrazoline-H), 5.70 (s, 1H, H-4), 6.20 (d, 1H,
H-2), 7.0 (d, 1H, H-1), 7.35–7.50 (m, 5H, Ar-H).
MS (EI): m/z 428 (15%) [M+]. Anal. Calcd for
C28H32N2O2: C,78.47; H, 7.53; N, 6.45. Found: C,
78.43; H, 7.49; N, 6.50.
3.1.5. Synthesis of (16R,50S)(8b,9a,14a)-3-Oxo-5a-and-
rostano[17,16-c]-10H-50-aryl-N-acetylpyrazoline (7a,b). A
mixture of the compounds (6a,b) (5 mmol) and acetyl
chloride (10 ml) was stirred overnight at room tempera-
ture. The excess acetyl chloride was evaporated under
reduced pressure, the residue was solidified with sodium

380
Abd El-Galil E. Amr et al. / Bioorg. Med. Chem. 14 (2006) 373–384
3.1.6.2. (16R,50S)(8b,9a,14a)-3-Oxo-androst-1,4-di-
tallized from the proper solvent to give N-substituted
pyrazoline derivatives (10a–d).
ene[17,16-c]-10H-50-(p-methoxy-phenyl)-N-acetylpyrazo-
25
line (8b). Yield 60%, mp: 217 ꢂC (MeOH); ½aꢂ +95 (c 1,
D
1
MeOH); IR (KBr): 1730, 1670, 1635 cmꢀ1. H NMR
(CDCl3): d 0.95 (s, 3H, CH3), 1.10 (s, 3H, CH3), 1.15–
1.20 (m, 1H, CH), 1.25–1.30 (m, 4H, CH2), 1.36–1.58
(m, 2H, CH2), 1.60–1.70 (m, 2H, CH2), 1.76 (m, 1H,
CH), 1.90–2.05 (m, 2H, CH2), 2.15 (s, 3H, COCH3),
2.30 (m, 1H, pyrazoline-H), 2.35–2.45 (m, 1H, CH),
3.40 (d, 1H, pyrazoline-H), 3.60 (s, 3H, OCH3), 5.75
(s, 1H, H-4), 6.20 (d, 1H, H-2), 7.10 (d, 1H, H-1),
7.30–7.55 (m, 4H, Ar-H). MS (EI): m/z 458 (15%)
[M+]. Anal. Calcd for C29H34N2O3: C, 75.95; H, 7.47;
N, 6.11. Found: C, 75.90; H, 7.42; N, 6.05.
3.1.8.1. (16R,50S)(8b,9a,14a)-3b-Trifluoroacetate-5a-
androstano[17,16-c]-10H-50-phenyl-N-methylpyrazoline (10a).
25
D
Yield 55%, mp: 251 ꢂC (EtOH/AcOEt); ½aꢂ +19 (c 1,
MeOH); IR (KBr): 1735, 1645 cmꢀ1
.
1H NMR
(CDCl3): d 0.75 (s, 3H, CH3), 0.85 (s, 3H, CH3),
0.92–1.10 (m, 1H, CH), 1.24–1.30 (m, 4H, 2CH2),
1.40–1.58 (m, 6H, 3CH2), 1.60–1.88 (m, 4H, 2CH2),
2.00–2.10 (m, 1H, CH), 2.20 (s, 3H, NCH3), 2.24–
2.36 (m, 2H, CH2), 2.40 (m, 1H, pyrazoline-H), 2.50
(m, 1H, CH), 2.55 (m, 1H, 3a-CH), 3.10 (m, 1H, 5a-
CH), 3.40 (d, 1H, pyrazoline-H), 7.20–7.45 (m, 5H,
Ar-H). MS (EI): m/z 502 (15%) [M+]. Anal. Calcd for
C29H37F3N2O2: C, 69.29; H, 7.42; N, 5.57. Found: C,
69.22; H, 7.38; N, 5.54.
3.1.7. Synthesis of (16R,50S)(8b,9a,14a)-3-oxo-androst-
1,4-diene[17,16-c]-10H-50-aryl-pyrazoline (9a,b). A solu-
tion of compounds (8a,b) (4 mmol) in 5% alcoholic
potassium hydroxide (15 ml) was refluxed for 2–3 h.
The reaction mixture was neutralized with hydrochloric
acid, the obtained solid was filtered off, washed with
water, dried, and crystallized from the proper solvent
to give compounds (9a,b).
3.1.8.2. (16R,50S)(8b,9a,14a)-3b-Trifluoroacetate-5a-
androstano[17,16-c]-10H-50-(p-methoxyphenyl)-N-methyl-
25
D
pyrazoline (10b). Yield 70%, mp: 315 ꢂC (AcOEt); ½aꢂ
+113 (c 1, MeOH); IR (KBr): 1745, 1640 cmꢀ1
.
1H
NMR (CDCl3): d 0.70 (s, 3H, CH3), 0.80 (s, 3H,
CH3), 0.90–1.00 (m, 1H, CH), 1.22–1.28 (m, 4H,
2CH2), 1.38–1.61 (m, 6H, 3CH2), 1.64–1.90 (m, 4H,
2CH2), 1.98–2.05 (m, 1H, CH), 2.30 (s, 3H, NCH3),
2.25–2.35 (m, 2H, CH2), 2.40 (m, 1H, pyrazoline-H),
2.45 (m, 1H, CH), 2.54 (m, 1H, 3a-CH), 3.05 (m,
1H, 5a-CH), 3.35 (d, 1H, pyrazoline-H), 3.60 (s, 3H,
OCH3), 7.30–7.35 (m, 4H, Ar-H). MS (EI): m/z 532
(15%) [M+]. Anal. Calcd for C30H39F3N2O3: C,
67.64; H, 7.38; N, 5.26. Found: C, 67.60; H, 7.34;
N, 5.22.
3.1.7.1. (16R,50S)(8b,9a,14a)-3-Oxo-androst-1,4-di-
ene[17,16-c]-10H-50-phenyl-pyrazoline (9a). Yield 70%,
25
mp: 284 ꢂC (MeOH); ½aꢂ +47 (c 1, MeOH); IR
D
1
(KBr): 3415, 1730 cmꢀ1. H NMR (DMSO-d6): d 0.70
(s, 3H, CH3), 0.90 (s, 3H, CH3), 0.95–1.05 (m, 1H,
CH), 1.24–1.32 (m, 4H, 2CH2), 1.38–160 (m, 2H,
CH2), 1.65–1.75 (m, 2H, CH2), 1.78 (m, 1H, CH),
1.85–2.00 (m, 2H, CH2), 2.30 (m, 1H, pyrazoline-H),
2.35–2.45 (m, 1H, CH), 3.45 (d, 1H, pyrazoline-H),
5.65 (s, 1H, H-4), 6.15 (d, 1H, H-2), 7.10 (d, 1H, H-1),
7.30–7.45 (m, 5H, Ar-H), 10.40 (s, 1H, NH, exchange-
able with D2O). MS (EI): m/z 386 (35%) [M+]. Anal.
Calcd for C26H30N2O: C, 80.79; H, 7.82; N, 7.25.
Found: C, 80.75; H, 7.78; N, 7.18.
3.1.8.3. (16R,50S)(8b, 9a,14a)-3b-Trifluoroacetate-5a-
androstano[17,16-c]-10H-50-phenyl-N-phenylpyrazoline (10c).
25
D
Yield 60, mp: 138 ꢂC (EtOH); ½aꢂ
+86 (c 1,
MeOH); IR (KBr): 1730, 1640 cmꢀ1
.
1H NMR
(CDCl3): d 0.70 (s, 3H, CH3), 0.85 (s, 3H, CH3),
0.92–0.98 (m, 1H, CH), 1.24–1.28 (m, 4H, 2CH2),
1.41–1.60 (m, 6H, 3CH2), 1.62–1.88 (m, 4H, 2CH2),
2.05–2.10 (m, 1H, CH), 2.24–2.30 (m, 2H, CH2),
2.35 (m, 1H, pyrazoline-H), 2.40 (m, 1H, CH), 2.55
(m, 1H, 3a-CH), 3.00 (m, 1H, 5a-CH), 3.50 (d,
1H, pyrazoline-H), 7.25–7.65 (m, 10H, Ar-H). MS
(EI): m/z 564 (35%) [M+]. Anal. Calcd for
C34H39F3N2O2: C, 72.31; H, 6.96; N, 4.96. Found:
C, 72.26; H, 6.92; H, 4.90.
3.1.7.2. (16R,50S)(8b,9a,14a)-3-Oxo-androst-1,4-di-
ene[17,16-c]-10H-50-(p-methoxy-phenyl)pyrazoline (9b).
25
D
Yield 65%, mp: 315 ꢂC (MeOH); ½aꢂ +135 (c 1,
MeOH); IR (KBr): 3410, 1725 cmꢀ1
.
1H NMR
(DMSO-d6): d 0.85 (s, 3H, CH3), 0.90 (s, 3H, CH3),
0.95–1.00 (m, 1H, CH), 1.22–1.31 (m, 4H, 2CH2),
1.44–162 (m, 2H, CH2), 1.66–1.70 (m, 2H, CH2), 1.75
(m, 1H, CH), 1.90–2.10 (m, 2H, CH2), 2.35 (m, 1H, pyr-
azoline-H), 2.40–2.50 (m, 1H, CH), 3.50 (d, 1H, pyrazo-
line-H), 3.60 (s, 3H, OCH3), 5.70 (s, 1H, H-4), 6.20 (d,
1H, H-2), 7.05 (d, 1H, H-1), 7.25–7.40 (m, 4H, Ar-H),
10.35 (s, 1H, NH, exchangeable with D2O). MS (EI):
m/z 416 (35%) [M+]. Anal. Calcd for C27H32N2O2: C,
77.84; H, 7.74; N, 6.73. Found: C, 77.80; H, 7.70; N,
6.68.
3.1.8.4. (16R,50S)(8b,9a,14a)-3b-Trifluoroacetate-5a-
androstano[17,16-c]-10H-50-(p-methoxyphenyl)-N-phenylpy-
razoline (10d). Yield 55%, mp: 158 ꢂC (EtOH); ½aꢂ25 +127
D
(c 1, MeOH); IR (KBr): 1740, 1635 cmꢀ1
.
1H NMR
(CDCl3): d 0.75 (s, 3H, CH3), 0.90 (s, 3H, CH3), 0.95–
1.00 (m, 1H, CH), 1.22–1.30 (m, 4H, 2CH2), 1.40–1.58
(m, 6H, 3CH2), 1.60–1.90 (m, 4H, 2CH2), 2.00–2.10
(m, 1H, CH), 2.25 (m, 1H, pyrazoline-H), 2.28–2.36
(m, 2H, CH2), 2.42 (m, 1H, CH), 2.50 (m, 1H, 3a-
CH), 3.05 (m, 1H, 5a-CH), 3.40 (d, 1H, pyrazoline-H),
3.65 (s, 3H, OCH3), 7.35–7.60 (m, 9H, Ar-H). MS
(EI): m/z 594 (45%) [M+]. Anal. Calcd for
C35H41F3N2O3: C, 70.68; H, 6.95; N, 4.71. Found: C,
70.62; H, 6.90; N, 4.67.
3.1.8. Synthesis of (16R,50S)(8b,9a,14a)-3b-trifluoroace-
tate-5a-androstano[17,16-c]-10H-50-aryl-N-substituted-pyr-
azoline (10a–d).
A mixture of the arylmethylene
derivatives (2a,b) (4 mmol) and hydrazine derivatives
(5 mmol), namely, methyl or phenyl hydrazines in gla-
cial acetic acid (15 ml) was refluxed for 5–7 h. The reac-
tion mixture was poured into ice water, the obtained
solid was filtered off, washed with water, dried, and crys-

Abd El-Galil E. Amr et al. / Bioorg. Med. Chem. 14 (2006) 373–384
381
3.1.9. Synthesis of (16R,50S)(8b,9a,14a)-3b-hydroxyl-5a-
androstano[17,16-c]-10H-50-aryl-N-substituted-pyrazoline
(11a–d). A solution of the fluoroacetate pyrazoline
derivatives (10a–d) (1 mmol) in 5% alcoholic KOH
(10 ml) was refluxed for ꢁ2 h, and then neutralized with
hydrochloric acid. The precipitated solid was filtered off,
washed with water, dried, and crystallized from the
proper solvent to give compounds (11a–d).
azoline-H), 2.46 (m, 1H, CH), 2.50 (m, 1H, 3a-CH), 3.05
(m, 1H, 5a-CH), 3.40 (d, 1H, pyrazoline-H), 3.60 (s, 3H,
OCH3), 7.30–7.80 (m, 9H, Ar-H), 10.20 (br s, 1H, OH,
exchangeable with D2O). MS (EI, 70 eV): m/z 498
(5%) [M+]. Anal. Calcd for C33H42N2O: C, 79.47; H,
8.49; N, 5.62. Found: C, 79.41; H, 8.43; N, 5.57.
3.1.10. Synthesis of (16R,50S)(8b,9a,14a)-3-oxo-5a-
androstano[17,16-c]-10H-50-aryl-N-substituted-pyrazoline
(12a–d). Using the same procedure for the synthesis of
derivative (6) using compounds (11a–d) as starting
materials.
3.1.9.1.
rostano[17,16-c]-10H-50-phenyl-N-methylpyrazoline (11a).
(16R,50S)(8b,9a,14a)-3b-Hydroxyl-5a-and-
25
Yield 65%, mp: 154 ꢂC (MeOH); ½aꢂ +96 (c 1, MeOH);
D
IR (KBr): 3515, 1650 cmꢀ1. 1H NMR (CDCl3): d 0.80 (s,
3H, CH3), 0.90 (s, 3H, CH3), 0.95–1.10 (m, 1H, CH),
1.25–1.30 (m, 4H, 2CH2), 1.40–1.60 (m, 6H, 3CH2),
1.65–1.90 (m, 4H, 2CH2), 1.95–2.00 (m, 1H, CH), 2.15
(s, 3H, NCH3), 2.30 (m, 1H, pyrazoline-H), 2.35–2.42
(m, 2H, CH2), 2.48 (m, 1H, CH), 2.54 (m, 1H, 3a-
CH), 3.05 (m, 1H, 5a-CH), 3.45 (d, 1H, pyrazoline-H),
7.25–7.45 (m, 5H, Ar-H), 10.55 (s, 1H, OH, exchange-
able with D2O). MS (EI): m/z 406 (18%) [M+]. Anal.
Calcd for C27H38N2O: C, 79.75; H, 9.42; N, 6.89.
Found: C, 79.70; H, 9.38; N, 6.84.
3.1.10.1. (16R,50S)(8b, 9a,14a)-3-Oxo-5a-androst-
ano[17,16-c]-10H-50-phenyl-N-methyl-pyrazoline (12a).
25
Yield 60%, mp: 312 ꢂC (EtOH); ½aꢂ +90 (c 1, MeOH);
D
IR (KBr): 1745, 1645 cmꢀ1. 1H NMR (CDCl3): d 0.80 (s,
3H, CH3), 0.92 (s, 3H, CH3), 0.95–1.00 (m, 1H, CH),
1.24–1.30 (m, 4H, 2CH2), 1.42–1.60 (m, 6H, 3CH2),
1.65–1.92 (m, 4H, 2CH2), 2.05 (m, 1H, CH), 2.10 (s,
3H, NCH3), 2.30 (m, 1H, pyrazoline-H), 2.32–2.42 (m,
2H, CH2), 2.48 (m, 1H, CH), 3.05 (m, 1H, 5a-CH),
3.35 (d, 1H, pyrazoline-H), 7.30–7.50 (m, 5H, Ar-H).
MS (EI): m/z 404 (28%) [M+]. Anal. Calcd for
C27H36N2O: C, 80.15; H, 8.97; N, 6.93. Found: C,
79.98; H, 8.94; N, 6.90.
3.1.9.2.
rostano[17,16-c]-10H-50-(p-methoxy-phenyl)-N-methylpy-
(16R,50S)(8b,9a,14a)-3b-Hydroxyl-5a-and-
25
razoline (11b). Yield 55%, mp: 257 ꢂC (AcOEt); ½aꢂ +43
D
(c 1, MeOH); IR (KBr): 3510, 1645 cmꢀ1
.
1H NMR
3.1.10.2.
ano[17,16-c]-10H-50-(p-methoxy-phenyl)-N-methylpyraz-
(16R,50S)(8b,9a,14a)-3-Oxo-5a-androst-
(CDCl3): d 0.80 (s, 3H, CH3), 0.95 (s, 3H, CH3), 1.00–
1.15 (m, 1H, CH), 1.24–1.32 (m, 4H, 2CH2), 1.38–1.62
(m, 6H, 3CH2), 1.65–1.88 (m, 4H, 2CH2), 1.95–2.00
(m, 1H, CH), 2.25 (s, 3H, NCH3), 2.28–2.38 (m, 2H,
CH2), 2.40 (m, 1H, pyrazoline-H), 2.46 (m, 1H, CH),
2.55 (m, 1H, 3a-CH), 3.10 (m, 1H, 5a-CH), 3.55 (d,
1H, pyrazoline-H), 3.75 (s, 3H, OCH3), 7.25–7.40 (m,
4H, Ar-H), 10.50 (br s, 1H, OH, exchangeable with
D2O). MS (EI): m/z 436 (10%) [M+]. Anal. Calcd for
C28H40N2O2: C, 77.02; H, 9.23; N, 6.42. Found: C,
76.96; H, 9.18; N, 6.37.
25
oline (12b). Yield 70%, mp: 287 ꢂC (AcOEt); ½aꢂ +73 (c
D
1, MeOH); IR (KBr): 1740, 1640 cmꢀ1
.
1H NMR
(DMSO-d6): d 0.75 (s, 3H, CH3), 0.85 (s, 3H, CH3),
0.90–1.05 (m, 1H, CH), 1.22–1.28 (m, 4H, 2CH2),
1.40–1.58 (m, 6H, 3CH2), 1.62–1.90 (m, 4H, 2CH2),
2.00 (m, 1H, CH), 2.10 (s, 3H, NCH3), 2.22–2.32 (m,
2H, CH2), 2.35 (m, 1H, pyrazoline-H), 2.46 (m, 1H,
CH), 3.00 (m, 1H, 5a-CH), 3.40 (d, 1H, pyrazoline-H),
3.65 (s, 3H, OCH3), 7.25–7.60 (m, 4H, Ar-H). MS
(EI): m/z 434 (18%) [M+]. Anal. Calcd for
C28H38N2O2: C, 77.37; H, 8.81; N, 6.45. Found: C,
77.30; H, 8.76; N, 6.40.
3.1.9.3.
rostano[17,16-c]-10H-50-phenyl-N-phenylpyrazoline (11c).
(16R,50S)(8b,9a,14a)-3b-Hydroxyl-5a-and-
25
Yield 72%, mp 181 ꢂC (MeOH); ½aꢂ +131 (c 1, MeOH);
3.1.10.3.
ano[17,16-c]-10H-50-phenyl-N-phenyl-pyrazoline
(16R,50S)(8b,9a,14a)-3-Oxo-5a-androst-
(12c).
D
IR (KBr): 3490, 1635 cmꢀ1. 1H NMR (CDCl3): d 0.75 (s,
3H, CH3), 0.85 (s, 3H, CH3), 0.90–1.00 (m, 1H, CH),
1.22–1.32 (m, 4H, 2CH2), 1.42–1.60 (m, 6H, 3CH2),
1.65–1.90 (m, 4H, 2CH2), 1.98–2.05 (m, 1H, CH), 2.25
(m, 1H, pyrazoline-H), 2.28–2.40 (m, 2H, CH2), 2.48
(m, 1H, CH), 2.50 (m, 1H, 3a-CH), 3.05 (m, 1H, 5a-
CH), 3.35 (d, 1H, pyrazoline-H), 7.15–7.45 (m, 10H,
Ar-H), 10.60 (br s, 1H, OH, exchangeable with D2O).
MS (EI): m/z 468 (26%) [M+]. Anal. Calcd for
C32H40N2O: C, 82.00; H, 8.60; N, 5.98. Found: C,
81.95; H, 8.20; N, 5.93.
25
Yield 75%, mp: 220 ꢂC (AcOEt); ½aꢂ +58 (c 1, MeOH);
D
1
IR (KBr): 1735, 1635 cmꢀ1. H NMR (DMSO-d6): d
0.80 (s, 3H, CH3), 0.85 (s, 3H, CH3), 0.90–1.00 (m,
1H, CH), 1.24–1.30 (m, 4H, 2CH2), 1.42–1.60 (m, 6H,
3CH2), 1.65–1.92 (m, 4H, 2CH2), 2.10 (m, 1H, CH),
2.24–2.40 (m, 2H, CH2), 2.45 (m, 1H, pyrazoline-H),
2.50 (m, 1H, CH), 3.00 (m, 1H, 5a-CH), 3.50 (d, 1H,
pyrazoline-H), 7.30–7.65 (m, 10H, Ar-H). MS (EI): m/
z 466 (35%) [M+]. Anal. Calcd for C32H38N2O: C,
82.36; H, 8.21; N, 6.00. Found: C, 82.26; H, 8.15; N,
5.95.
3.1.9.4. (16R,50S)(8b,9a,14a)-3b-Hydroxyl-5a-andro
stano[17,16-c]-10H-50-(p-methoxy-phenyl)-N-phenylpyra
3.1.10.4.
ano[17,16-c]-10H-50-(p-methoxy-phenyl)-N-phenylpyrazo-
(16R,50S)(8b,9a,14a)-3-Oxo-5a-androst-
25
D
zoline (11d). Yield 75%, mp: 275 ꢂC (MeOH); ½aꢂ +120
25
line (12d). Yield 65%, mp: 266 ꢂC (AcOEt); ½aꢂ +88 (c
(c 1, MeOH); IR (KBr): 3520, 1640 cmꢀ1
.
1H NMR
D
(CDCl3): d 0.80 (s, 3H, CH3), 0.85 (s, 3H, CH3), 0.90–
0.98 (m, 1H, CH), 1.24–1.28 (m, 4H, 2CH2), 1.40–1.60
(m, 6H, 3CH2), 1.65–1.7.85 (m, 4H, 2CH2), 2.00–2.10
(m, 1H, CH), 2.20–2.32 (m, 2H, CH2), 2.35 (m, 1H, pyr-
1, MeOH); IR (KBr): 1745, 1640 cmꢀ1
.
1H NMR
(CDCl3): d 0.75 (s, 3H, CH3), 0.85 (s, 3H, CH3), 0.95–
1.05 (m, 1H, CH), 1.24–1.32 (m, 4H, 2CH2), 1.40–1.60
(m, 6H, 3CH2), 1.62–1.90 (m, 4H, 2CH2), 1.98 (m, 1H,

382
Abd El-Galil E. Amr et al. / Bioorg. Med. Chem. 14 (2006) 373–384
CH), 2.30 (m, 1H, pyrazoline-H), 2.32–2.40 (m, 2H,
CH2), 2.48 (m, 1H, CH), 3.10 (m, 1H, 5a-CH), 3.40
(d, 1H, pyrazoline-H), 3.70 (s, 3H, OCH3), 7.25–7.65
(m, 9H, Ar-H). MS (EI): m/z 496 (5%) [M+]. Anal. Calcd
for C33H40N2O2: C, 79.79; H, 8.12; N, 5.64. Found: C,
79.72; H, 8.00; N, 5.60.
(m, 2H, CH2), 1.60–1.78 (m, 2H, CH2), 1.38–1.92 (m,
1H, CH), 1.95–2.10 (m, 2H, CH2), 2.25 (m, 1H, pyrazo-
line-H), 2.28–2.35 (m, 1H, CH), 3.45 (d, 1H, pyrazoline-
H), 3.70 (s, 3H, OCH3), 5.75 (s, 1H, H-4), 6.30 (d, 1H,
H-2), 7.0 (d, 1H, H-1), 7.35–7.70 (m, 9H, Ar-H). MS
(EI): m/z 492 (5%) [M+]. Anal. Calcd for C32H36N2O2:
C, 80.45; H, 7.37; N, 5.69. Found: C, 80.40; H, 7.32;
N, 5.62.
3.1.11. Synthesis of (16R,50S)(8b,9a,14a)-3-oxo-androst-
1,4-diene[17,16-c]-10H-50-aryl-N-substituted-pyrazoline
(13a–d). Using the same procedure for the synthesis of
compounds (8) using compounds (12a–d) as starting
materials.
3.1.12. Synthesis of 3(Z)-ethylidene androstano[17,16-c]-
10H-50-aryl-N-substituted-pyrazoline (14a–d). To a stir-
red solution of ethylidene triphenylphosphorane
(1.2 mmol) in dimethylsulfoxide (100 ml), cycloketone
derivatives (12a–d) (1 mmol) in dry benzene (60 ml) were
added dropwise, and then heated at 60 ꢂC for 10–12 h.
The reaction mixture was cooled, poured into ice water,
the formed product was extracted with ethyl acetate,
dried over anhydrous sodium sulfate, and evaporated
under reduced pressure. The obtained residue was crys-
tallized from the proper solvent to give compounds
(14a-d).
3.1.11.1. (16R,50S)(8b,9a,14a)-3-Oxo-androst-1,4-di-
(13a).
ene[17,16-c]-10H-50-phenyl-N-methylpyrazoline
25
Yield 62%, mp: 271 ꢂC (EtOH); ½aꢂ +35 (c 1, MeOH);
D
1
IR (KBr): 1747, 1642 cmꢀ1. H NMR (DMSO-d6): d
0.75 (s, 3H, CH3), 0.90 (s, 3H, CH3), 0.95–1.00 (m,
1H, CH), 1.25–1.30 (m, 4H, 2CH2), 1.40–1.60 (m, 2H,
CH2), 1.65–1.78 (m, 2H, CH2), 1.38–1.92 (m, 1H,
CH), 1.95–2.15 (m, 2H, CH2), 2.20 (s, 3H, NCH3),
2.30 (m, 1H, pyrazoline-H), 2.35–2.42 (m, 1H, CH),
3.35 (d, 1H, pyrazoline-H), 5.85 (s, 1H, H-4), 6.35 (d,
1H, H-2), 7.10 (d, 1H, H-1), 7.35–7.65 (m, 5H, Ar-H).
MS (EI): m/z 400 (15%) [M+]. Anal. Calcd for
C27H32N2O: C, 80.95; H, 8.05; N, 7.00. Found: C,
80.88; H, 7.98; N, 6.94.
3.1.12.1. 3(Z)-Ethylidene androstano[17,16-c]-10H-50-
phenyl-N-methylpyrazoline (14a). Yield 68%, mp: 237 ꢂC
25
(MeOH); ½aꢂ +73 (c 1, MeOH); IR (KBr): 1642 cmꢀ1
.
D
1H NMR (CDCl3): d 0.80 (s, 3H, CH3), 0.95 (s, 3H,
CH3), 0.98–1.05 (m, 1H, CH), 1.24–1.29 (m, 4H,
2CH2), 1.40–1.60 (m, 6H, 3CH2), 1.70 (d, 3H, CH3),
1.74–1.90 (m, 4H, 2CH2), 2.10 (m, 1H, CH), 2.15 (s,
3H, NCH3), 2.20–2.32 (m, 2H, CH2), 2.35 (m, 1H, pyr-
azoline-H), 2.48 (m, 1H, CH), 3.10 (m, 1H, 5a-CH), 3.40
(d, 1H, pyrazoline-H), 5.45 (q, 1H, CH@C), 7.25–7.55
(m, 5H, Ar-H). MS (EI): m/z 416 (5%) [M+]. Anal. Calcd
for C29H40N2: C, 83.59; H, 9.68; N, 6.73. Found: C,
83.52; H, 9.64; N, 6.68.
3.1.11.2. (16R,50S)(8b,9a, 14a)-3-Oxo-androst-1,4-di-
ene[17,16-c]-10H-50-(p-methoxy-phenyl)-N-methylpyrazo-
25
D
line (13b). Yield 74%, mp: 206 ꢂC (EtOH); ½aꢂ +170 (c
1, MeOH); IR (KBr): 1745, 1635 cmꢀ1
.
1H NMR
(DMSO-d6): d 0.70 (s, 3H, CH3), 0.80 (s, 3H, CH3),
0.90–0.98 (m, 1H, CH), 1.24–1.29 (m, 4H, 2CH2),
1.42–1.60 (m, 2H, CH2), 1.62–1.76 (m, 2H, CH2),
1.36–1.92 (m, 1H, CH), 1.94–2.14 (m, 2H, CH2), 2.25
(s, 3H, NCH3), 2.28–2.38 (m, 1H, CH), 2.40 (m, 1H,
pyrazoline-H), 3.40 (d, 1H, pyrazoline-H), 3.60 (s, 3H,
OCH3), 5.80 (s, 1H, H-4), 6.25 (d, 1H, H-2), 6.95 (d,
1H, H-1), 7.40–7.70 (m, 4H, Ar-H). MS (EI): m/z 430
(48%) [M+]. Anal. Calcd for C28H34N2O2: C, 78.10; H,
7.96; N, 6.51. Found: C, 77.96; H, 7.91; N, 6.45.
3.1.12.2. 3(Z)-Ethylidene androstano[17,16-c]-10H-50-
(p-methoxyphenyl)-N-methyl-pyrazoline (14b). Yield
25
56%, mp: 289 ꢂC (EtOH); ½aꢂ +117 (c 1, MeOH); IR
D
1
(KBr): 1645 mꢀ1. H NMR (DMSO-d6): d 0.75 (s, 3H,
CH3), 0.85 (s, 3H, CH3), 0.95–1.00 (m, 1H, CH), 1.22–
1.35 (m, 4H, 2CH2), 1.42–1.58 (m, 6H, 3CH2), 1.60–
1.72 (m, 4H, 2CH2), 1.75 (d, 3H, CH3), 3.05 (m, 1H,
CH), 2.25 (s, 3H, NCH3), 2.30 (m, 1H, pyrazoline-H),
2.32–2.40 (m, 2H, CH2), 2.50 (m, 1H, CH), 3.10 (m,
1H, 5a-CH), 3.50 (d, 1H, pyrazoline-H), 3.65 (s, 3H,
OCH3), 5.40 (q, 1H, CH@C), 7.30–7.70 (m, 4H, Ar-
H). MS (EI): m/z 446 (35%) [M+]. Anal. Calcd for
C30H42N2O: C, 80.66; H, 9.48; N, 6.27. Found: C,
80.60; H, 9.42; N, 6.23.
3.1.11.3. (16R,50S)(8b,9a, 14a)-3-Oxo-androst-1,4-di-
(13c).
ene[17,16-c]-10H-50-phenyl-N-phenylpyrazoline
25
Yield 65%, mp: 155 ꢂC (EtOH); ½aꢂ +25 (c 1, MeOH);
D
1
IR (KBr): 1740, 1640 cmꢀ1. H NMR (DMSO-d6): d
0.75 (s, 3H, CH3), 0.85 (s, 3H, CH3), 0.95–1.05 (m,
1H, CH), 1.25–1.31 (m, 4H, 2CH2), 1.44–1.62 (m, 2H,
CH2), 1.62–1.78 (m, 2H, CH2), 1.38–1.92 (m, 1H,
CH), 1.95–2.15 (m, 2H, CH2), 2.18–2.32 (m, 1H, CH),
2.35 (m, 1H, pyrazoline-H), 3.30 (d, 1H, pyrazoline-
H), 5.70 (s, 1H, H-4), 6.20 (d, 1H, H-2), 7.05 (d, 1H,
H-1), 7.35–7.75 (m, 10H, Ar-H). MS (EI): m/z 462
(25%) [M+]. Anal. Calcd for C32H34N2O: C, 83.07; H,
7.41; N, 6.06. Found: C, 82.92; H, 7.36; N, 5.98.
3.1.12.3. 3(Z)-Ethylidene androstano[17,16-c]-10H-50-
phenyl-N-phenylpyrazoline (14c). Yield 66%, mp:
25
D
293 ꢂC (MeOH); ½aꢂ +46 (c 1, MeOH); IR (KBr):
1640, 1600 cmꢀ1
.
1H NMR (DMSO-d6): d 0.75 (s,
3H, CH3), 0.85 (s, 3H, CH3), 0.90–1.00 (m, 1H,
CH), 1.24–1.32 (m, 4H, 2CH2), 1.42–1.60 (m, 6H,
3CH2), 1.68 (d, 3H, CH3), 1.70–1.95 (m, 4H, 2CH2),
2.10 (m, 1H, CH), 2.30 (m, 1H, pyrazoline-H),
2.35–2.45 (m, 2H, CH2), 2.50 (m, 1H, CH), 3.10 (m,
1H, 5a-CH), 3.50 (d, 1H, pyrazoline-H), 5.50 (q,
1H, CH@C), 7.30–7.65 (m, 10H, Ar-H). MS (EI):
m/z 478 (5%) [M+]. Anal. Calcd for C34H42N2: C,
3.1.11.4. (16R,50S)(8b,9a,14a)-3-Oxo-androst-1,4-di-
ene[17,16-c]-10H-50-(p-methoxy-phenyl)-N-phenylpyrazo-
25
D
line (13d). Yield 74%, mp: 174 ꢂC (MeOH); ½aꢂ +41 (c
1, MeOH); IR (KBr): 1740, 1635 cmꢀ1
.
1H NMR
(CDCl3): d 0.80 (s, 3H, CH3), 0.90 (s, 3H, CH3), 0.95–
1.10 (m, 1H, CH), 1.22–1.28 (m, 4H, 2CH2), 1.42–1.60

Abd El-Galil E. Amr et al. / Bioorg. Med. Chem. 14 (2006) 373–384
383
85.30; H, 8.84; N, 5.85. Found: C, 85.00; H, 8.79; N,
5.81.
solution in dimethyl sulfoxide (DMSO). Control ani-
mals received intraperitoneal injections of DMSO.
For each concentration and control, 10 female rats
were injected with the tested compounds for viability
twice daily for two weeks. From the mortality data
of all tested animals, the intraperitoneal LD50 and
LD90 values for each agent were determined according
to Austen and Brocklehurst.31
3.1.12.4. 3(Z)-Ethylidene androstano[17,16-c]-10H-50-
(p-methoxyphenyl)-N-phenyl-pyrazoline (14d). Yield
25
62%, mp: 263 ꢂC (MeOH); ½aꢂ +69 (c 1, MeOH); IR
D
1
(KBr): 1645, 1610 mꢀ1. H NMR (DMSO-d6): d 0.70
(s, 3H, CH3), 0.90 (s, 3H, CH3), 0.95–1.05 (m, 1H,
CH), 1.24–1.33 (m, 4H, 2CH2), 1.42–1.60 (m, 6H,
3CH2), 1.70 (d, 3H, CH3), 1.74–1.95 (m, 4H, 2CH2),
2.00 (m, 1H, CH), 2.30 (m, 1H, pyrazoline-H), 2.34–
2.40 (m, 2H, CH2), 2.50 (m, 1H, CH), 3.10 (m, 1H,
5a-CH), 3.35 (d, 1H, pyrazoline-H), 3.60 (s, 3H,
OCH3), 5.45 (q, 1H, CH@C), 7.30–7.80 (m, 9H, Ar-
H). MS (EI): m/z 508 (40%) [M+]. Anal. Calcd for
C35H44N2O: C, 82.63; H, 8.72: N, 5.51. Found: C,
82.58; H, 8.68; N, 5.45.
References and notes
1. Amr, A. E.; Abdulla, M. M. Indian J. Heterocycl. Chem.
2002, 12, 129.
2. Amr, A. E.; Hegab, M. I.; Ibrahim, A. A.; Abdalah, M.
M. Monatschefte fur Chemie 2003, 134, 1395.
3. Amr, A. E.; Abou-Ghalia, M. H. Amino Acids 2004, 26,
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3.2. Antiandrogenic activity in female rats
3.2.1. Aim and rational. Neuman and Elger described28
the method for testing the antiandrogenic activity in
ovariectomized female rats. The protection of the anti-
androgen Cyperoteroneꢀ against the trophic effect of
testosterone on uterine and prenuptial growth was
equally studied in intact as well as castrated female
rats.
3.2.2. Procedure. Female Sprague–Dawley rats weight-
ing 40–45 g were overotomized as described above.
After one week, animals were divided into 38 groups
(12 animals) of which 36 received new compounds, a
group received control, and a group received Cyperoter-
oneꢀ. The treatment was continued over a period of 12
days with daily subcutaneous injection of 0.3 mg testos-
terone propionate and the same dose of each individual
new tested compound. The control received only testos-
terone propionate. On the 13th day, the animals were
sacrificed, and the uteri and perpetual glands were
weighed. The increase in female accessory sexual organs
due to testosterone treatment was dose-despondently re-
duced by antiandrogen.29,30
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Dose–response curves were established for various
doses of the antiandrogen at a given dose of testoster-
one propionate or for various doses of testosterone
propionate at a given dose of the antiandrogen. From
the dose–response curve, the relative antiandrogenic
potency of the newly synthesized compounds was cal-
culated and compared to that of Cyperoteroneꢀ as a
reference drug.
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3.2.3. Determination of acute to xicity (LD50 and LD90).
Female Sprague–Dawley albino rats were used to
determine intraperitoneal LD50 and LD90 of the tested
compounds. Prior to the determination of LD50 and
LD90 values, a range finding screen was conducted
using 20 rats treated with each tested compound at
dose ranging from 3 to 2000 mg/kg/dose level. Based
on the mortality observed within 14 days, the doses
used for the LD50 and LD90 determination were 3,
10, 30, 100, 300, 1000, and 2000 mg/kg of compound
administered by intraperitoneal injection as a 10%

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