Heterocyclic ring extension of estrone: Synthesis and cytotoxicity of fused pyran, pyrimidine and thiazole derivatives

Article abstract of DOI:10.1016/j.steroids.2014.03.012

The one pot reaction of estrone with the aromatic aldehydes 2a-c and either of malononitrile or ethyl cyanoacetate afforded the fused pyran derivatives 4a-f. On the other hand, carrying the same reaction using thiourea instead of the cyanomethylene reagent gave the fused pyrimidine derivatives 6a-c. The latter compounds reacted with phenacyl bromide to give the thiazolo[3,2-a] pyrimidine derivatives 8a-c. The reaction of the title compound with bromine gave the monobromo derivative 13 which in turn reacted with either thiourea or cyanothioacetamide to give the thiazole derivatives 14 and 16, respectively. The cytotoxicity of the newly synthesized products was evaluated against six human cancer and normal cell lines where the results showed that compounds 4c, 4f, 6b, 8b, 8c, 10, 13, 16, 18c and 19c exhibited optimal cytotoxic effect against the cancer cell lines, with IC₅₀'s in the nM range.

Full text of DOI:10.1016/j.steroids.2014.03.012

Steroids 84 (2014) 46–56
Contents lists available at ScienceDirect
Steroids
journal homepage: www.elsevier.com/locate/steroids
Heterocyclic ring extension of estrone: Synthesis and cytotoxicity
of fused pyran, pyrimidine and thiazole derivatives
b
c
Rafat M. Mohareb a, , Fatima Al-Omran , Rasha A. Azzam
⇑
a Department of Chemistry, Faculty of Science, Cairo University, Giza, A.R., Egypt
b Department of Chemistry, Kuwait University, P.O. Box 5969, 13060 Safat, Kuwait
c Department of Chemistry, Faculty of Science, Helwan University, Cairo, A.R., Egypt
a r t i c l e i n f o
a b s t r a c t
Article history:
The one pot reaction of estrone with the aromatic aldehydes 2a–c and either of malononitrile or ethyl
cyanoacetate afforded the fused pyran derivatives 4a–f. On the other hand, carrying the same reaction
using thiourea instead of the cyanomethylene reagent gave the fused pyrimidine derivatives 6a–c. The
latter compounds reacted with phenacyl bromide to give the thiazolo[3,2-a]pyrimidine derivatives
8a–c. The reaction of the title compound with bromine gave the monobromo derivative 13 which in turn
reacted with either thiourea or cyanothioacetamide to give the thiazole derivatives 14 and 16, respec-
tively. The cytotoxicity of the newly synthesized products was evaluated against six human cancer and
normal cell lines where the results showed that compounds 4c, 4f, 6b, 8b, 8c, 10, 13, 16, 18c and 19c
exhibited optimal cytotoxic effect against the cancer cell lines, with IC50’s in the nM range.
Ó 2014 Elsevier Inc. All rights reserved.
Received 25 September 2013
Received in revised form 21 February 2014
Accepted 15 March 2014
Available online 28 March 2014
Keywords:
Estrone
Pyran
Pyrimidine
Thiazole
Cytotoxicity
1. Introduction
present paper, we report an efficient synthesis of estrone possess-
ing pyran, pyrimidine and thiazole ring systems. This study fo-
Several D-ring alkylated estrone analogues display exception-
ally high affinity for estrogen receptors [1–3]. In particular, com-
pounds in which an E-ring is formed are known to be involved in
the inhibition of steroidogenic enzymes [4,5]. Such compounds
also have an effect on steroid dehydrogenase activity and the
ability to inhibit the detrimental action of the steroid sulfatase
enzyme [6–8]. Generally, E-ring extended steroids have been
accessed by modification of the C17-ketone in the D-ring by
either arylimine or oximino formation [9–11], addition of a car-
bon nucleophile [12] or hydrazone formation [13–15]. Other ap-
proaches have included ketone reduction, silyl enol ether
formation or ring-closing metathesis (giving five- or six mem-
bered E-rings) [16,17]. Chemical modification of the steroid
D-ring provides a way to alter the functional groups, sizes and
stereochemistry of the D-ring, and numerous structure–activity
relationships have been established by such synthetic alterations.
Steroids bearing heterocycles fused to the D-ring of the steroid
nucleus have been of pharmaceutical interest [18–21]. In the
cused on the synthesis of heterocyclic estrone derivatives
together with their cytotoxic evaluations towards human cancer
and normal cell lines.
2. Experimental
2.1. Synthetic methods, analytical and spectral data
The starting steroid, estrone, was purchased from Sigma Com-
pany, USA. All solvents were dried by distillation prior to using.
Melting points were recorded on Buchi melting point apparatus
D-545; 13C NMR and 1H NMR spectra were recorded on Bruker
DPX200 instrument in DMSO with TMS as internal standard for
protons and solvent signals as internal standard for carbon spectra.
Chemical shift values are mentioned in d (ppm). Mass spectra
were recorded on EIMS (Shimadzu) and ESI-esquire 3000
Bruker Daltonics instrument. Elemental analyses were carried out
by the Microanalytical Data Unit Ludwig-Maximilians-Universitat-
Muenchen, Germany. The progress of all reactions was monitored
by TLC on 2 ꢀ 5 cm pre-coated silica gel 60 F254 plates of thickness
of 0.25 mm (Merck).
⇑
Corresponding author. Fax: +20 2 35676570.
E-mail address: raafat_mohareb@yahoo.com (R.M. Mohareb).
http://dx.doi.org/10.1016/j.steroids.2014.03.012
0039-128X/Ó 2014 Elsevier Inc. All rights reserved.

R.M. Mohareb et al. / Steroids 84 (2014) 46–56
47
2.1.1. (6bS, 8aS,13aS,13bR)-10-amino-4-hydroxy-8a-methyl-12-phen
yl-1,2,6b,7,8,8a-12,13,13a,13b-decahydronaphtho[20,10:4,5]indeno[1,2-
b]pyran-11-carbonitrile (4a) (6bS, 8aS,13aS,13bR)-4,10-dihydroxy-8a-
methyl-12-phenyl-1,2,6b,7,8,8a-12,13,13a,13b-decahydronaphtho[20,
10: 4,5] indeno[1,2-b]pyran-11-carbonitrile (4b), (6bS, 8aS,13aS,13bR)-
10-amino-8a-methyl-12-(4-chlorophenyl)-4-hydroxy-8a-methyl-1,2,
6b,7,8,8a-12,13,13a,13b-decahydronaphtho[20,10:4,5] indeno[1,2-b]
pyran-11-carbonitrile (4c), (6bS, 8aS,13aS,13bR)-4,10-dihydroxy-8a-
methyl-12-(4-chlorophenyl)-8a-methyl-1,2,6b,7,8,8a-12,13,13a,13b-
decahydronaphtho [20,10:4,5] indeno[1,2-b]pyran-11-carbonitrile (4d)
(6bS, 8aS,13aS,13bR)-10-amino-8a-methyl-12-(4-methoxyphenyl)-4-
hydroxy-8a-methyl-1,2,6b,7,8,8a-12,13,13a,13b-decahydronaphtho [20,
10:4,5] indeno[1,2-b]pyran-11-carbonitrile (4e), (6bS, 8aS,13aS,13bR) -
4,10-dihydroxy-8a-methyl-12-(4-methoxyphenyl)-8a-methyl-1,2,6b, 7,
8,8a-12,13,13a,13b-decahydronaphtho [20,10:4,5] indeno[1,2-b]pyran-
11-carbonitrile (4f)
General procedure: To a solution of estrone (0.270 g, 1 mmol) in
absolute ethanol (40 mL) containing triethylamine (0.025 mL),
either of malononitrile (0.066 g, 1 mmol) or ethyl cyanoacetate
(0.113 g, 1 mmol) and either of the aromatic aldehydes namely
benzaldehyde (106 g, 1 mmol), 4-chlorobenzaldehyde (0.140 g,
1 mmol) or 4-methoxybenzaldehyde (0.136 g, 1 mmol) were
added. The reaction mixture, in each case, was heated under reflux
for 1 h and the formed solid product produced from the hot solu-
tion was collected by filtration. Thin layer chromatography re-
vealed just a single spot which proved the presence of a single
product.
Compound 4a: HPLC purity = 90% (C-18 NovaPak column;
MeOH:H2O/70:30), tr = 24 min; pale yellow crystals from
EtOAc:hexane (88%), m.p. 199–202 °C; IR (KBr) cmꢁ1: 3580–3403,
3055, 2938, 2220, 1638, 1560; 1H-NMR (DMSO): d 0.89 (s, 3H),
1.30–1.60 (m, 5H), 1.69 (dt, 3H, J = 7.0, 2.8 Hz), 1.72–1.83 (m,
2H), 1.88–2.02 (m, 1H), 2.90–2.97 (m, 2H), 2.99–3.12 (m, 1H),
4.44 (s, 2H, D2O exchangeable), 5.77 (s, 1H), 6.44 (d, 1H,
J = 2.3 Hz,), 6.76 (dd, 1H, J = 8.2 Hz and J = 2.3 Hz), 6.88–7.36 (m,
5H), 9.02 (s, 1H, D2O exchangeable); 13C-NMR (DMSO): d 13.8,
22.8, 23.3, 26.9, 31.6, 38.6, 45.6, 49.6, 112.6, 117.8, 120.3, 124.2,
126.3, 128.6, 129.0, 135.4, 136.9, 140.3, 142.6, 146.8, 148.5,
149.5, 154.3, 155.9, 160.3, 196.6. MS: m/e = 424 (M+, 29%); Analysis
Calcd for C28H28N2O2: C, 79.22; H, 6.65; N, 6.60%. Found: C, 79.41;
H, 6.83; N, 6.49%.
ClN2O2: C, 73.27; H, 5,93; N, 6.10; Cl, 7.72%. Found: C, 73,22; H,
6.05; N, 6.37; Cl, 7.99%.
Compound 4d: HPLC purity = 90% (C-18 NovaPak column;
MeOH:H2O/85:15), tr = 23 min; pale yellow crystals from
EtOAc:hexane (86%), m.p. 140–142 °C; IR (KBr) cmꢁ1: 3544–3426,
3053, 2939, 2220, 1634, 1560; 1H-NMR (DMSO): d 0.82 (s, 3H),
1.32–1.56 (m, 5H), 1.67 (dt, 3H, J = 7.0, 2.8 Hz), 1.70–1.84 (m,
2H), 1.88–2.03 (m, 1H), 2.91–2.97 (m, 2H), 2.98–3.11 (m, 1H),
5.70 (s, 1H), 6.42 (d, 1H, J = 3.0 Hz), 6.67 (dd, 1H, J = 8.0 Hz,
J = 3.0 Hz), 6.86–7.40 (m, 4H), 8.06, 9.05 (2s, 2H, D2O exchange-
able); 13C-NMR (DMSO): d 14.3, 19.8, 22.7, 23.0, 26.9, 27.8, 31.5,
38.8, 40.4, 45.2, 112.6, 117.2, 120.8, 123.8, 125.2, 129.3, 135.4,
137.2, 140.8, 143.0, 146.8, 154.3, 155.9, 159.7, 160.8, 196.3. MS:
m/e = 459 (M+, 36%); Analysis Calcd for C28H26ClNO3: C, 73.11; H,
5,70; N, 3.05; Cl, 7.71%. Found: C, 73.38; H, 5.44; N, 6.94; Cl, 7.93%.
Compound 4e: HPLC purity = 93% (C-18 NovaPak column;
MeOH:H2O/90:10), tr = 20 min; pale yellow crystals from
EtOAc:hexane (84%), m.p. 188–190 °C; (IR (KBr) cmꢁ1: 3532–
3422, 3058, 2932, 2223, 1636, 1563; 1H-NMR (DMSO): d 0.86 (s,
3H), 1.31–1.59 (m, 5H), 1.66 (dt, 3H, J = 7.0, 2.8 Hz,), 1.71–1.83
(m, 2H), 1.87–2.02 (m, 1H), 2.90–2.96 (m, 2H), 2.98–3.11 (m,
1H), 3.18 (s, 3H), 4.43 (s, 2H, D2O exchangeable), 5.71 (s, 1H),
6.44 (d, 1H, J = 3.8 Hz), 6.62 (dd, 1H, J = 6.8 Hz, J = 3.8 Hz), 6.83–
7.43 (m, 4H), 9.02, 9.32 (2s, 2H, D2O exchangeable); 13C-NMR
(DMSO): d 14.6, 19.4, 22.7, 23.5, 26.9, 27.9, 31.3, 38.8, 40.6, 45.4,
49.7, 54.6, 112.2, 116.4, 121.4, 124.2, 125.6, 128.4, 130.4, 133.2,
136.4, 140.4, 142.9, 146.8, 148.6, 154.6, 159.9, 160.8. MS:
m/e = 454 (M+, 16%); Analysis Calcd for C29H30N2O3: C, 76.63.11;
H, 6.65; N, 6.16%. Found: C, 76.82; H, 6.73; N, 6.29%.
Compound 4f: HPLC purity = 88% (C-18 NovaPak column;
MeOH:H2O/90:10), tr = 24 min; pale yellow crystals from
EtOAc:hexane (73%), m.p. 233–235 °C; IR (KBr) cmꢁ1: 3548–3432,
3053, 2935, 2220, 1638, 1570; 1H-NMR (DMSO): d 0.83 (s, 3H),
1.30–1.62 (m, 5H), 1.67 (dt, 3H, J = 7.0, 2.8 Hz), 1.72–1.85 (m,
2H), 1.86–2.02 (m, 1H), 2.90–2.96 (m, 2H), 2.98–3.11 (m, 1H),
3.14 (s, 3H), 5.71 (s, 1H), 6.48 (d, 1H, J = 3.3 Hz), 6.70 (dd, 1H,
J = 8.2 Hz and J = 3.3 Hz), 6.88–7.45 (m, 4H), 8.16 (s, 1H, D2O
exchangeable); 13C-NMR (DMSO): d 14.2, 19.3, 22.9, 23.5, 26.9,
27.9, 31.5, 38.4, 40.8, 44.6, 49.5, 54.6, 112.3, 116.8, 122.6, 124.5,
125.8, 129.1, 130.6, 132.8, 134.2, 141.2, 143.4, 146.8, 148.6,
153.2, 155.9, 160.2. MS: m/e = 455 (M+, 32%); Analysis Calcd for
Compound 4b: HPLC purity = 92% (C-18 NovaPak column;
MeOH:H2O/70:30), tr = 22 min; pale yellow crystals from
EtOAc:hexane (90%), m.p. 168–169 °C; IR (KBr) cmꢁ1: 3540–3423,
3057, 2932, 2222, 1637, 1563; 1H-NMR (DMSO): d 0.84 (s, 3H),
1.33–1.84 (m, 5H), 1.70 (dt, 3H, J = 6.89, 3.0 Hz), 1.72–1.85 (m,
2H), 1.88–1.22 (m, 1H), 2.88–2.94 (m, 2H), 2.96–3.11 (m, 1H),
5.73 (s, 1H), 6.44 (d, 1H, J = 2.3 Hz), 6.78 (dd, 1H, J = 7.8 Hz and
J = 2.3 Hz), 6.85–7.38 (m, 5H), 9.03 (s, 1H, D2O exchangeable);
13C-NMR (DMSO): d 14.6, 19.8, 22.6, 23.3, 26.9, 27.3, 31.5, 38.8,
40.2, 45.6, 49.8, 112.4, 116.9, 120.5, 125.9, 129.3, 135.4, 137.2,
140.3, 142.6, 146.8, 148.8, 155.9, 159.4, 160.1, 196.8. MS:
m/e = 425 (M+, 28%); Analysis Calcd for C28H27NO3: C, 79.03; H,
6.40; N, 3.29%. Found: C, 79.18; H, 6.64; N, 3.33%.
Compound 4c: HPLC purity = 93% (C-18 NovaPak column;
MeOH:H2O/70:30), tr = 18 min; pale yellow crystals from
EtOAc:hexane (88%), m.p. 180–182 °C; IR (KBr) cmꢁ1: 3535–3433,
3056, 2936, 2224, 1639, 1562; 1H-NMR (DMSO): d 0.82 (s, 3H),
1.37–1.58 (m, 5H), 1.67 (dt, 3H, J = 7.0, 2.8 Hz,), 1.72–1.82 (m,
2H), 1.86–2.03 (m, 1H), 2.90–2.94 (m, 2H), 2.96–3.09 (m, 1H),
4.60 (s, 2H, D2O exchangeable), 5.68 (s, 1H), 6.45 (d, 1H,
J = 2.2 Hz), 6.60 (dd, 1H, J = 6.89 Hz, J = 2.2 Hz), 6.88–7.41 (m, 4H),
9.06 (s, 1H, D2O exchangeable); 13C-NMR (DMSO): d 14.4, 19.7,
22.8, 23.3, 26.9, 27.3, 31.8, 38.6, 40.5, 45.3, 49.3, 110.6, 117.2,
120.6,123.9, 126.4, 130.6, 135.6, 141.3, 143.8, 148.0, 154.8, 155.4,
159.3, 160.6. MS: m/e = 458 (M+, 33%); Analysis Calcd for C28H27
C29H29NO4: C, 76.46.11; H, 6.42; N, 3.07%. Found: C, 76.51; H,
6.53; N, 2.99%.
2.1.2. 6bS,8aS,13aS,13bR)-10-Mercapto-8a-methyl-12-phenyl-2,6b,7,
8,8a,9,12,13,13a,13b-decahydro-1H-naphtho[20,10:4,5]indeno[1,2-d]
pyrimidin-4-ol (6a), 6bS,8aS,13aS,13bR)-10-Mercapto-8a-methyl-12-
(4-chlorophenyl)-2,6b,7,8,8a,9,12,13,13a,13b-decahydro-1H-naphtho
[20,10:4,5] indeno[1,2-d]pyrimidin-4-ol (6b) and 6bS,8aS,13aS,13bR)-
10-Mercapto-8a-methyl-12-(4-methoxyphenyl)-2,6b,7,8,8a,9,12,13,
13a,13b-decahydro-1H-naphtho [20,10:4,5] indeno[1,2-d]pyrimidin-4-
ol (6c)
General procedure: To a solution of estrone (0.270 g, 1 mmol) in
absolute ethanol (40 mL) containing triethylamine (0.025 mL),
either of the aromatic aldehydes namely benzaldehyde (106 g,
1 mmol), 4-chlorobenzaldehyde (0.140 g, 1 mmol) or 4-methoxy-
benzaldehyde (0.136 g, 1 mmol) together with thiourea
(0.76 g,1 mmol) were added. The reaction mixture, in each case,
was heated under reflux for 4 h, and the formed solid product pro-
duced from the hot solution was collected by filtration. Thin layer
chromatography revealed just a single spot which proved the pres-
ence of a single product.
Compound 6a: HPLC purity = 91% (C-18 NovaPak column;
MeOH:H2O/90:10), tr = 20 min; white crystals from EtOAc:
hexane (83%), m.p. 222–225 °C; IR (KBr) cmꢁ1: 3532–3421, 3058,
2938, 1622, 1560; 1H-NMR (DMSO): d 0.83 (s, 3H), 1.03 (s, 1H),

48
R.M. Mohareb et al. / Steroids 84 (2014) 46–56
1.32–1.57 (m, 5H), 1.68 (dt, J = 7.2, 2.9 Hz, 3H), 1.70–1.85 (m, 2H),
1.83–2.02 (m, 1H), 2.92–2.97 (m, 2H), 2.98–3.11 (m, 1H), 5.73 (s,
1H), 6.42 (d, 1H, J = 2.5 Hz), 6.60 (dd, 1H, J = 7.23 Hz and
J = 2.5 Hz), 6.93–7.42 (m, 5H), 8.22, 9.03 (2s, 2H, D2O exchange-
able); 13C-NMR (DMSO): d 13.9, 19.4, 22.8, 23.3, 26.9, 27.3, 31.6,
38.6, 40.2, 45.8, 60.3, 112.6, 113.8, 121.9, 124.8, 126.4, 127.2,
129.5, 135.4, 146.8, 153.0 154.8, 162.9. MS: m/e = 416 (M+, 42%);
Analysis Calcd for C26H28N2OS: C, 74.96; H, 6.77; N, 6.72; S, 7.70%.
Found: C, 75.72; H, 6.89; N, 6.66; S, 7.93%.
162.7. MS: m/e = 516 (M+, 37%); Analysis Calcd for C34H32N2OS: C,
79.03; H, 6.24; N, 5.42; S, 6.21%. Found: C, 78.88; H, 6.37; N,
5.28; S, 6.07%.
Compound 8b: HPLC purity = 73% (C-18 NovaPak column;
MeOH:H2O/80:20), tr = 19 min; white crystals from EtOAc:hexane
(82%), m.p. 222–225 °C; IR (KBr) cmꢁ1: 3544–3423, 3052, 2935,
1630, 1563; 1H-NMR (DMSO): d 0.83 (s, 3H), 1.32–1.58 (m, 5H),
1.67 (dt, J = 6.9, 2.5 Hz, 3H), 1.70–1.88 (m, 2H), 1.80–2.03 (m,
1H), 2.90–2.93 (m, 2H), 2.96–3.01 (m, 2H), 6.45 (d, 1H,
J = 3.9 Hz), 6.63 (dd, 1H, J = 6.99 Hz, J = 3.9 Hz), 6.50 (s, 1H), 6.92–
7.40 (m, 9H), 9.04 (s, 1H, D2O exchangeable); 13C-NMR (DMSO):
d 14.0, 19.8, 22.8, 23.7, 26.9, 27.8, 31.6, 38.6, 40.1, 45.8, 57.3,
60.8, 104.2, 122.9, 123.8, 124.0, 125.9, 126.8, 128.5, 129.5, 130.3,
135.6, 142.0, 142.6, 143.1, 146.2, 149.6, 154.0, 154.8, 162.9. MS:
m/e = 551 (M+, 42%); Analysis Cald for C34H31ClN2OS: C, 74.09; H,
5.67; N, 6.43; S, 5.82%. Found: C, 73.87; H, 5.72; N, 4.88; S, 6.08%.
Compound 8c: HPLC purity = 88% (C-18 NovaPak column;
MeOH:H2O/87:13), tr = 20 min; pale yellow crystals from EtOAc:
hexane (80%), m.p. 188–190 °C; IR (KBr) cmꢁ1: 3538–3453, 3058,
2928, 1625, 1562; 1H-NMR (DMSO): d 0.83 (s, 3H), 1.35–1.59 (m,
5H), 1.68 (dt, J = 6.8, 2.7 Hz, 3H), 1.71–1.86 (m, 2H), 1.80–2.01
(m, 1H), 2.90–2.96 (m, 2H), 2.97–3.01 (m, 2H), 3.11 (s, 3H), 6.42
(d, 1H, J = 3.8 Hz), 6.52 (s, 1H), 6.64 (dd, 1H, J = 7.38 Hz and
J = 3.8 Hz), 6.84–7.42 (m, 9H), 9.05 (s, 1H, D2O exchangeable);
13C-NMR (DMSO): d 14.2, 19.6, 21.8, 22.6, 23.3, 25.4, 26.6, 27.9,
31.6, 38.8, 40.2, 45.7, 50.9, 60.3, 112.6, 119.3, 122.6, 123.6, 124.8,
126.2, 126.8, 129.9, 133.6, 138.6, 141.8, 142.3, 143.0, 146.6,
151.9, 153.3, 163.2. MS: m/e = 530 (M+, 38%); Analysis Cald for C35
H34N2OS: C, 79.21; H, 6.46; N, 5.28; S, 6.04%. Found: C, 79.03; H,
6.55; N, 6.37; S, 5.93%.
Compound 8d: HPLC purity = 88% (C-18 NovaPak column;
MeOH:H2O/87:13), tr = 17 min; pale yellow crystals from EtOAc:
hexane (77%), m.p. 122–124 °C; IR (KBr) cmꢁ1: 3538–3453, 3058,
2928, 1625, 1562; 1H-NMR (DMSO): d 0.83 (s, 3H), 1.35–1.59 (m,
5H), 1.67 (dt, J = 6.9, 2.5 Hz, 3H), 1.70–1.88 (m, 2H), 1.80–2.03
(m, 1H), 2.90–2.96 (m, 2H), 2.85–3.01 (m, 2H), 3.21(s, 3H), 6.40
(d, 1H, J = 2.6 Hz), 6.53 (s, 1H), 6.63 (dd, 1H, J = 8.72 Hz and
J = 2.6 Hz), 6.81–7.40 (m, 9H), 9.06 (s, 1H, D2O exchangeable);
13C-NMR (DMSO): d 14.4, 19.8, 22.6, 23.3, 26.8, 27.9, 30.5, 31.6,
38.8, 40.6, 44.6, 50.5, 55.3, 60.8, 112.8, 120.8, 122.8, 123.4, 124.2,
126.8, 127.3, 129.9, 133.8, 134.2, 138.9, 141.4, 142.3, 143.0,
147.3, 152.3, 154.6, 163.1. MS: m/e = 546 (M+, 24%); Analysis Calcd
for C35H34N2O2S: C, 76.89; H, 6.27; N, 5.12; S, 5.86%. Found: C,
76.93; H, 6.38; N, 5.06; S, 5.66%.
Compound 6b: HPLC purity = 83% (C-18 NovaPak column;
MeOH:H2O/80:20), tr = 21 min; white crystals from EtOAc: hexane
(86%), m.p. 183–186 °C; IR (KBr) cmꢁ1: 3548–3433, 3054, 2933,
1631, 1562; 1H-NMR (DMSO): d 0.85 (s, 3H), 1.04 (s, 1H), 1.30–
1.59 (m, 5H), 1.67 (dt, J = 7.1, 2.7 Hz, 3H), 1.71–1.87 (m, 2H),
1.85–2.04 (m, 1H), 2.91–2.96 (m, 2H), 2.98–3.09 (m, 1H), 5.70 (s,
1H), 6.44 (d, 1H, J = 2.8 Hz), 6.66 (dd, 1H, J = 6.94 Hz and
J = 2.8 Hz), 6.82–7.40 (m, 4H), 8.06, 9.05 (2s, 2H, D2O exchange-
able); 13C-NMR (DMSO): d 14.2, 19.6, 22.8, 23.3, 26.9, 27.6, 31.6,
38.6, 40.3, 45.8, 60.5, 112.8, 114.0, 124.8, 126.8, 129.5, 135.4,
138.8, 146.9, 147.3, 152.3, 154.3, 162.9. MS: m/e = 451 (M+, 40%);
Analysis Calcd for C26H27ClN2OS: C, 69.24; H, 6.03; Cl, 7.86; N,
6.21; S, 7.11%. Found: C, 69.03; H, 6.33; Cl, 9.05; N, 6.39; S, 7.30%.
Compound 6c: HPLC purity = 89% (C-18 NovaPak column;
MeOH:H2O/87:13), tr = 18 min; white crystals from EtOAc: hexane
(88%), m.p. 203–205 °C; IR (KBr) cmꢁ1: 3528–3412, 3052, 2930,
1622, 1560; 1H-NMR (DMSO): d 0.86 (s, 3H), 1.04 (s, 1H), 1.32–
1.61 (m, 5H), 1.66 (dt, J = 7.0, 2.7 Hz, 3H), 1.71–1.85 (m, 2H),
1.83–2.01 (m, 1H), 2.90–2.96 (m, 2H), 2.96–3.11 (m, 1H), 3.11 (s,
3H), 5.69 (s, 1H), 6.40 (d, 1H, J = 3.4 Hz), 6.60 (dd, 1H, J = 8.31 Hz
and J = 3.4 Hz), 6.77–7.38 (m, 4H), 8.04, 9.05 (2s, 2H, D2O
exchangeable); 13C-NMR (DMSO): d 14.0, 19.4, 22.6, 23.0, 26.9,
27.8, 31.6, 38.6, 40.6, 45.7, 56.4, 60.2, 112.8, 113.8, 122.3, 126.8,
129.7, 138.6, 142.8, 146.4, 148.3, 151.8, 152.6, 154.3, 163.0. MS:
m/e = 446 (M+, 24%); Analysis Calcd for C27H30N2O2S: C, 72.61; H,
6.77; N, 6.27; S, 7.18%. Found: C, 72.88; H, 6.83; N, 6.18; S, 7.44%.
2.1.3. (6bS,8aS,15aS,15R)-8a-Methyl-10,14-diphenyl-1,2,6b,7,8,8a,14,
15,15a,15b-decahydronaphtho[20,10:4,5]indeno[2,1-e]thiazolo[3,2-a]
pyrimidin-4-ol (8a), (6bS,8aS,15aS,15R)-10-(4-chlorophenyl)-8a-methyl-
14-phenyl-1,2,6b,7,8,8a,14,15,15a,15b-decahydronaphtho [20,10:4,5] inde-
no[2,1-e]thiazolo[3,2-a]pyrimidin-4-ol (8b), (6bS,8aS,15aS,15R)-
10-(4-methylphenyl)-8a-Methyl-14-phenyl–1,2,6b,7,8,8a,14,15,15a,
15b-decahydronaphtho [20,10:4,5] indeno[2,1-e]thiazolo[3,2-a]pyrimid-
in-4-ol (8c), (6bS,8aS,15aS,15R)-10-(4-methoxyphenyl)-8a-Methyl-
14-phenyl–1,2,6b,7,8,8a,14,15,15a,15b-decahydronaphtho [20,10:4,5]
indeno[2,1-e]thiazolo[3,2-a]pyrimidin-4-ol (8d)
General procedure: To a solution of compound 6a (0.416 g,
1 mmol) in ethanol (40 mL), either 2-bromo-1-phenylethanone
(0.20 g, 1 mmol), 2-bromo-1-(4-chlorophenyl)ethanone (0.235 g,
1 mmol), 2-bromo-1-(4-methylphenyl)ethanone (0.215 g, 1 mmol)
or 2-bromo-1-(4-methoxyphenyl)ethanone (0.247 g, 1 mmol) was
added. The reaction mixture, in each case was heated under reflux
for 3 h then evaporated under vacuum. The remaining product was
triturated with diethyl ether and the formed solid product was col-
lected by filtration.
Compound 8a: HPLC purity = 87% (C-18 NovaPak column;
MeOH:H2O/90:10), tr = 18 min; white crystals from EtOAc:hexane
(80%), m.p. 130–133 °C; IR (KBr) cmꢁ1: 3562–3441, 3054, 2936,
1631, 1562; 1H-NMR (DMSO): d 0.82 (s, 3H), 1.31–1.60 (m, 5H),
1.68 (dt, J = 7.1, 2.9 Hz, 3H), 1.71–1.87 (m, 2H), 1.81–2.01 (m,
1H), 2.90–2.94 (m, 2H), 2.96–3.03 (m, 2H), 6.44 (d, 1H,
J = 3.2 Hz), 6.64 (dd, 1H, J = 8.2 Hz and J = 3.2 Hz), 6.51 (s, 1H),
7.29–7.42 (m, 10H), 9.05 (s, 1H, D2O exchangeable); 13C-NMR
(DMSO): d 13.4, 19.2, 22.8, 23.6, 26.9, 27.3, 31.6, 38.6, 40.4, 45.2,
57.3, 60.2, 104.8, 12.3, 122.4, 123.6, 124.4, 124.9, 125.9, 126.8,
128.5, 129.5, 135.4, 141.8, 142.6, 143.1, 149.4, 154.3, 154.8,
2.1.4. (6bS,8aS,12aS,12bR)-4-hydroxy,8a-methyl-9-phenyl-6b,7,8,8a,
9,12,12a,12b-octahydro-1H-naphtho[20,10:4,5]indeno[1,2-d]thiazole-
10(2H)-thione (10)
To a solution of estrone (0.270 g, 1 mmol) in absolute ethanol
(40 mL) containing triethylamine (0.025 mL) elemental sulphur
(0.32 g, 1 mmol) and phenylisothiocyanate (1.30 g, 1 mmol) were
added. The reaction mixture was heated under reflux for 2 h till a
solid product being formed and the latter was collected by
filtration.
Compound 10: HPLC purity = 75% (C-18 NovaPak column;
MeOH:H2O/80:20), tr = 18 min; pale yellow crystals from EtOAc:
hexane (80%), m.p. 193–195 °C; IR (KBr) cmꢁ1: 3520–3432, 3056,
2930, 1640, 1566, 1200–1195; 1H-NMR (CDCl3): d 0.80 (s, 3H),
1.34–1.61 (m, 5H), 1.68 (dt, J = 7.0, 2.9 Hz, 3H), 1.70–1.86 (m,
2H), 1.83–2.02 (m, 1H), 2.86–2.90 (m, 2H), 2.92–3.08 (m, 2H),
6.63 (dd, 1H, J = 7.5 Hz and J = 3.2 Hz), 7.31–7.42 (m, 5H), 9.04 (s,
1H); 13C-NMR (CDCl3): d 14.0, 19.6, 21.6, 22.5, 23.9, 25.4, 31.4,
38.3, 41.3, 44.3, 50.7, 54.0, 112.3, 118.7, 120.8, 129.2, 130.8,
133.2, 142.3, 143.0, 149.2, 152.3, 154.4, 183.2. MS: m/e = 419
(M+, 32%); Analysis Calcd for C25H25NOS2: C, 71.56; H, 6.01; N,
3.34; S, 15.28%. Found: C, 71.49; H, 5.87; N, 3.44; S, 15.06%.

R.M. Mohareb et al. / Steroids 84 (2014) 46–56
49
2.1.5. (6bS,8aS,12aS,12bR)-10-hydrazono-8a-methyl-9-phenyl-2,6b,7,
8,8a,9,10,12,12a,12b-decahydro-1H-naphtho[20,10:4,5]indeno[1,2-d]-
thiazol-4-ol (12a) and (6bS,8aS,12aS,12bR)- 8a-methyl-9-phenyl-10
(2-phenylhydrazono)-2,6b,7,8,8a,9,10,12,12a,12b-decahydro-1H-nap
htho[20,10:4,5]indeno[1,2-d]thiazol-4-ol (12a)
General procedure: To a solution of compound 10 (0.419 g,
1 mmol) in 1,4-dioxan (40 mL), either hydrazine hydrate
(0.50 mL, 1 mmol) or phenylhydrazine (0.109 g, 1 mmol) was
added. The reaction mixture was heated under reflux for 3 h then
evaporated under vacuum. The remaining product was triturated
with diethyl ether and the formed solid product was collected by
filtration.
Compound 14: HPLC purity = 81% (C-18 NovaPak column;
MeOH:H2O/90:10), tr = 20 min; yellow crystals from acetone
(83%), m.p. 220–223 °C; IR (KBr) cmꢁ1: 3560–3325, 3053, 2923,
1633, 1532; 1H-NMR (DMSO): d 0.84 (s, 3H), 1.33–1.56 (m, 5H),
1.64 (dt, J = 7.1, 2.9 Hz, 3H), 1.71–1.88 (m, 2H), 2.87–2.93 (m,
2H), 2.96–3.04 (m, 2H), 4.28 (s, 2H, D2O exchangeable), 6.35 (d,
1H, J = 2.8 Hz), 6.63 (dd, 1H, J = 7.8 Hz and J = 2.8 Hz), 9.03 (s, 1H,
D2O exchangeable); 13C-NMR (DMSO): d 14.2, 24.3, 25.8, 26.6,
29.8, 31.4, 38.4, 43.6, 44.8, 48.8, 120.3, 125.8, 128.6, 130.5, 134.7,
140.3, 154.7, 156.6, 160.4. MS: m/e = 326 (M+, 55%); Analysis Calcd
for C19H22N2OS: C, 69.90; H, 6.79; N, 8.58; S, 9.82%. Found: C,
69.84; H, 6.82; N, 8.47; S, 10.03%.
Compound 12a: HPLC purity = 80% (C-18 NovaPak column;
MeOH:H2O/85:15), tr = 23 min; pale yellow crystals from EtOAc:
hexane (84%), m.p. 204–207 °C; IR (KBr) cmꢁ1: 3568–3412, 3053,
2935, 1660, 1638, 1562; 1H-NMR (CDCl3): d 0.82 (s, 3H), 1.32–
1.58 (m, 5H), 1.69 (dt, J = 6.8, 3.2 Hz, 3H), 1.68–1.82 (m, 2H),
2.84–2.92 (m, 2H), 2.95–3.06 (m, 2H), 4.88 (s, 2H, D2Oexchange-
able), 6.37 (d, 1H, J = 2.7 Hz), 6.62 (dd, 1H, J = 8.4 and J = 2.7 Hz),
7.31–7.46 (m, 5H), 9.03 (s, 1H); 13C-NMR (CDCl3): d 13.8, 19.8,
21.6, 22.7, 23.9, 25.6, 31.6, 38.2, 41.6, 44.6, 51.6, 54.0, 115.9,
119.9, 124.8, 128.7, 130.3, 142.6, 144.6, 149.2, 152.3, 156.2,
172.1. MS: m/e = 417 (M+, 44%); Analysis Calcd for C25H27N3OS: C,
71.91; H, 6.52; N, 10.06; S, 7.68%. Found: C, 72.27; H, 6.79; N,
9.84; S, 7.92%.
Compound 12b: HPLC purity = 72% (C-18 NovaPak column;
MeOH:H2O/87:13), tr = 21 min; yellow crystals from EtOAc:MeOH
(73%), m.p. 189–192 °C; IR (KBr) cmꢁ1: 3551–3442, 3056, 2937,
1660, 1634, 1560; 1H-NMR (CDCl3): d 0.80 (s, 3H), 1.33–1.60 (m,
5H), 1.66 (dt, J = 7.2, 2.7 Hz, 3H), 1.70–1.88 (m, 2H), 2.87–2.91
(m, 2H), 2.95–3.06 (m, 2H), 6.34 (d, 1H, J = 4.2 Hz), 6.62 (dd, 1H,
J = 7.6 Hz and J = 4.2 Hz), 7.28–7.36 (m, 10H), 8.22 (s, 1H, D2O
exchangeable), 9.05 (s, 1H, D2O exchangeable); 13C-NMR (CDCl3):
d 13.9, 19.5, 21.6, 22.9, 23.9, 25.8, 31.6, 38.2, 41.4, 44.6, 51.6,
54.3, 115.3, 120.3, 123.4, 123.9, 125.3, 128.4, 133.8, 134.7, 140.6,
142.8, 143.8, 149.0, 152.6, 156.5, 172.1. MS: m/e = 493 (M+,
100%); Analysis Calcd for C31H31N3OS: C, 75.42; H, 6.33; N, 8.51;
S, 6.50%. Found: C, 75.63; H, 6.50; N, 8.31; S, 6.73%.
2.1.8. 2-((6bS,8aS,12aS,12bR)-4-Hydroxy-8a-methyl-2,6b,7,8,8a,12,
12a,12b-octahydro-1H-naphtho[20,10:4,5]indeno[1,2-d]thiazol-10-yl)
acetonitrile (16)
To a solution of compound 6a (0.416 g, 1 mmol) in ethanol
(40 mL), cyanothioacetamide (0.10 g, 1 mmol) was added. The
reaction mixture was heated under reflux for 4 h then evaporated
under vacuum. The remaining product was triturated with diethyl
ether and the formed solid product was collected by filtration.
Compound 16: HPLC purity = 85% (C-18 NovaPak column;
MeOH:H2O/82:18), tr = 22 min; pale yellow crystals from
EtOAc:hexane (88%), m.p. 288–290 °C; IR (KBr) cmꢁ1: 3558–3422,
3053, 2932, 2220, 1642, 1560; 1H-NMR (CDCl3): d 0.82 (s, 3H),
1.33–1.60 (m, 5H), 1.63 (dt, J = 7.3, 3.3 Hz, 3H), 1.70–1.88 (m,
2H), 2.87–2.95 (m, 2H), 2.91–2.99 (m, 2H), 4.99 (s, 2H), 6.43 (d,
1H, J = 3.4 Hz), 6.60 (dd, 1H, J = 8.5 Hz and J = 3.4 Hz), 9.02 (s, 1H,
D2O exchangeable); 13C-NMR (CDCl3): d 14.1, 19.9, 21.9, 22.8,
23.5, 27.9, 30.8, 31.6, 38.8, 40.9, 44.3, 50.5, 54.2, 116.8, 124.2,
126.8, 129.9, 142.3, 143.0, 154.6, 163.0. MS: m/e = 350 (M+,
100%); Analysis Calcd for C21H22N2OS: C, 71.97; H, 6.33; N, 7.99;
S, 9.15%. Found: C, 72.29; H, 6.58; N, 8.21; S, 9.28%.
2.1.9. (6bS,8aS,12aS,12bR,Z)-4-hydroxy-8a-methyl-N0-phenyl-2,6b,7,
8,8a,12,12a,12b-octahydro-1H-naphtho[20,10:4,5]indeno[1,2-d]triazo-
le-10-carbohydrazonoyl cyanide(18a), (6bS,8aS,12aS,12bR,Z)-4-hydr-
oxy-8a-methyl-N0-(p-tolyl)-2,6b,7,8,8a,12,12a,12b-octahydro-1H-nap-
htho [20,10:4,5] indeno[1,2-d]triazole-10-carbohydrazonoyl cyanide
(18b), (6bS,8aS,12aS,12bR,Z)-4-hydroxy-8a-methyl-N0-(4-chlorophen-
yl)-2,6b,7,8,8a,12,12a,12b-octahydro-1H-naphtho [20,10:4,5] indeno[1,
2-d]triazole-10-carbohydrazonoyl cyanide (18c) and (6bS,8aS,12aS,
12bR,Z)-4-hydroxy-8a-methyl-N0-(4-methoxyphenyl)-2,6b,7,8,8a,12,
12a,12b-octahydro-1H-naphtho [20,10:4,5] indeno[1,2-d]triazole-10-
carbohydrazonoyl cyanide (18d)
General procedure: To a solution of compound 16 (0.350 g,
1 mmol) in ethanol (30 mL) containing sodium acetate (2.5 g),
either of benzenediazonium chloride (0.01 mol), 4-meth-
ylbenzenediazonium chloride (1 mmol), 4-chlorobenzenediazoni-
um chloride (1 mmol) or 4-methoxybenzenediazonium chloride
[prepared by adding sodium nitrite solution (0.007 g, 1 mmol) to
a cold solution of the appropriate aniline or its derivative (1 mmol)
in concentrated hydrochloric acid (3 mL, 18 N) with continuous
stirring] was added with stirring. The reaction mixture was kept
at room temperature for 1 h and the formed solid product, in each
case, was collected by filtration.
Compound 18a: HPLC purity = 88% (C-18 NovaPak column;
MeOH:H2O/85:15), tr = 20 min; orange crystals from 1,4-dioxan
(78%), m.p. 172–174 °C; IR (KBr) cmꢁ1: 3548–3430, 3056, 2928,
2223, 1633, 1562; 1H-NMR (DMSO): d 0.83 (s, 3H), 1.31–1.61 (m,
5H), 1.62 (dt, J = 7.1, 3.1 Hz, 3H), 1.71–1.89 (m, 2H), 2.84–2.95
(m, 2H), 2.97–3.02 (m, 2H), 6.42 (d, 1H, J = 2.9 Hz), 6.62 (dd, 1H,
J = 6.5 Hz and J = 2.9 Hz), 7.03–7.38 (m, 5H), 8.22 (s, 1H, D2O
exchangeable), 9.05 (s, 1H, D2O exchangeable); 13C-NMR (DMSO):
d 14.0, 19.7, 21.7, 22.9, 27.6, 31.6, 38.8, 40.9, 44.3, 50.5, 116.6,
120.3, 122.6, 124.8, 126.4, 130.6, 135.2, 138.3, 140.8, 142.3,
2.1.6. (8R,9S,13S,14S)-16-bromo-3-hydroxy-13-methyl-7,8,9,11,12,
13,15,16-octahydro-6H-cyclopenta[a]phenanthen-17(14H)-one (13)
A solution of estrone (0.270 g, 1 mmol) in glacial acetic acid
(40 mL) was heated to 60 °C then a solution of bromine (0.160 g,
1 mmol) in acetic acid (5 mL) was added drop wise with continu-
ous stirring. The reaction mixture was stirred at 60 °C for an addi-
tional 1 h and the solid product, so formed, upon pouring onto ice/
water was collected by filtration.
Compound 13: HPLC purity = 77% (C-18 NovaPak column;
MeOH:H2O/88:12), tr = 19 min; yellow crystals from acetone
(89%), m.p. 189–192 °C; IR (KBr) cmꢁ1: 3542–3385, 3056, 1720,
2920, 1631, 1538; 1H-NMR (CDCl3): d 0.89 (s, 3H), 1.30–1.60 (m,
5H), 1.68 (dt, J = 7.0, 2.7 Hz, 3H), 1.71–1.86 (m, 2H), 2.90–2.96
(m, 2H), 2.98–3.10 (m, 2H), 5.04 (s, 1H), 6.34 (d, 1H, J = 2.8 Hz),
6.60 (dd, 1H, J – 8.4 Hz and J = 2.8 Hz), 9.05 (s, 1H, D2O exchange-
able); 13C-NMR (CDCl3): d 13.9, 23.9, 25.5, 26.6, 29.8, 31.6, 38.2,
43.2, 44.6, 46.2, 47.2, 48.6, 122.3, 128.4, 134.7, 154.8, 156.5,
180.6. MS: m/e = 349 (M+, 28%); Analysis Calcd for C18H21BrO2: C,
61.90; H, 6.06; Br, 22.88%. Found: C, 62.11; H, 6.29; Br, 23.17%.
2.1.7. (6bS,8aS,12aS,12bR)-10-amino-8a-methyl-2,6b,7,8,8a,12,12a,
12b-octahydro-1H-naphtho[20,10:4,5]indeno[1,2-d]thiazol-4-ol (14)
To a solution compound 13 (0.349 g, 1 mmol) in absolute etha-
nol (40 mL), thiourea (0.76 g, 1 mmol) was added. The reaction
mixture was heated under reflux for 3 h then left to cool. The
formed solid product was collected by filtration dried and moni-
tored through TLC for the purity.

50
R.M. Mohareb et al. / Steroids 84 (2014) 46–56
143.0, 149.4, 152.6, 163.0, 165.8. MS: m/e = 454 (M+, 44%); Analysis
Calcd for C27H26N4OS: C, 71.34; H, 5.76; N, 12.32; S, 7.05%. Found:
C, 71.49; H, 5.92; N, 12.19; S, 6.88%.
1670, 1632, 1560; 1H-NMR (DMSO): d 0.82 (s, 3H), 1. .30–1.58
(m, 5H), 1.61 (dt, J = 7.0, 3.1 Hz, 3H), 1.72–1.89 (m, 2H), 2.82–
2.88 (m, 2H), 2.92–2.95 (m, 2H), 6.40 (d, 1H, J = 3.3 Hz), 6.62 (dd,
1H, J = 8.2 Hz and J = 3.3 Hz), 7.26–7.44 (m, 10H), 8.22, (s, 1H,
D2O exchangeable), 9.03 (s, 1H, D2O exchangeable); 13C-NMR
(DMSO): d 14.2, 19.5, 21.9, 22.5, 23.7, 27.3, 30.4, 31.7, 38.3, 41.1,
44.3, 50.5, 119.3, 121.4, 122.6, 124.3, 125.7, 126.4, 128.0, 129,8,
130.8, 133.2, 135.0, 140.8, 142.6, 143.4, 149.2, 152.8, 163.3,
166.0, 168.3, 179.3 MS: m/e = 589 (M+, 28%); Analysis Cald for C34-
H31N5OS2: C, 69.24; H, 5.30; N, 11.87; S, 10.87%. Found: C, 69.44; H,
5.52; N, 11.64; S, 10.73%.
Compound 19b: HPLC purity = 87% (C-18 NovaPak column;
MeOH:H2O/90:10), tr = 19 min; red crystals from ethanol (80%),
m.p. 189–191 °C; IR (KBr) cmꢁ1: 3545–3427, 3053, 2932, 1677,
1631, 1560; 1H-NMR (DMSO): d 0.83 (s, 3H), 1.30–1.55 (m, 5H),
1.60 (dt, J = 7.2, 2.6 Hz, 3H), 1.72–1.90 (m, 2H), 2.82–2.89 (m,
2H), 2.91–2.96 (m, 2H), 3.09 (s, 3H), 6.45 (d, 1H, J = 3.2 Hz), (dd,
1H, J = 7.6 Hz and J = 3.2 Hz), 7.28–7.43 (m, 9H), 8.24 (s, 1H, D2O
exchangeable), 9.04 (s, 1H, D2O exchangeable); 13C-NMR (DMSO):
d 14.1, 19.5, 21.6, 22.9, 23.4, 23.7, 24.6, 28.4, 30.9, 31.7, 38.5,
40.9, 44.8, 50.6, 120.4, 123.9, 124.4, 125.3, 126.6, 128.3, 130.6,
133.6, 135.0, 138.5, 140.6, 142.2, 143.6, 148.8, 152.7, 154.6,
163.0, 166.3, 172.5, 180.3. MS: m/e = 603 (M+, 28%); Analysis Calcd
for C35H33N5OS2: C, 69.62; H, 5.51; N, 11.60; S, 10.62%. Found: C,
69.82; H, 5.73; N, 11.53; S, 10.85%.
Compound 19c: HPLC purity = 89% (C-18 NovaPak column;
MeOH:H2O/90:10), tr = 22 min; yellow crystals from ethanol
(83%), m.p. 122–124 °C; IR (KBr) cmꢁ1: 3555–3441, 3056, 2940,
1666, 1635, 1563; 1H-NMR (DMSO): d 0.82 (s, 3H), 1.31–1.59 (m,
5H), 1.63 (dt, J = 7.2, 3.0 Hz, 3H), 1.72–1.89 (m, 2H), 2.81–2.88
(m, 2H), 2.92–2.97 (m, 2H), 6.46 (d, 1H, J = 3.2 Hz), 6.62 (dd, 1H,
J = 8..1 Hz and J = 3.2 Hz), 7.31–7.44 (m, 9H), 8.26 (s, 1H, D2O
exchangeable), 9.04 (s, 1H, D2O exchangeable); 13C-NMR (DMSO):
d 13.8, 19.5, 21.7, 22.8, 23.7, 28.5, 30.5, 38.8, 41.6, 44.6, 50.6,
119.4, 120.6, 122.8, 123.7, 124.3, 125.6, 127.4, 128.9, 133.9,
134.2, 138.4, 142.3, 145.8, 148.8, 153.2, 155.1, 162.3, 166.4,
172.4, 180.4. MS: m/e = 624 (M+, 28%); Analysis Calcd for C34H30-
ClN5OS2: C, 65.42; H, 4.84; Cl, 5.68; N, 11.22; S, 10.27%. Found: C,
64.53; H, 5.08; Cl, 5.82; N, 11.36; S, 10.36%.
Compound 19d: HPLC purity = 89% (C-18 NovaPak column;
MeOH:H2O/85:15), tr = 19 min; pale yellow crystals from ethanol
(73%), m.p. > 300 °C; IR (KBr) cmꢁ1: 3532–3451, 3056, 2925,
1635, 1558; 1H-NMR (DMSO): d 0.82 (s, 3H), 1.28–2.82 (m, 5H),
1.60 (dt, J = 6.8, 3.3 Hz, 3H), 1.702–1.89 (m, 2H), 2.81–2.87 (m,
2H), 2.90–2.93 (m, 2H), 3.26 (s, 3H), 6.37 (d, 1H, J = 3.1 Hz), 6.63
(dd, 1H, J = 6.3 Hz and J = 3.1 Hz), 7.31–7.46 (m, 9H), 8.23 (s, 1H,
D2O exchangeable), 9.02 (s, 1H, D2O exchangeable); 13C-NMR
(DMSO): d 14.0, 19.7, 21.9, 22.3, 23.6, 27.6, 31.1, 38.6, 41.2, 44.8,
51.4, 54.3, 119.8, 120.3, 122.7, 123.7, 127.2, 129.2, 135.0, 138.7,
141.5, 142.8, 143.1, 149.7, 153.6, 154.9, 163.4, 166.4, 172.0,
180.2. MS: m/e = 619 (M+, 26%); Analysis Calcd for C35H33N5O2S2:
C, 67.82; H, 5.37; N, 11.30; S, 10.35%. Found: C, 67.68; H, 5.49; N,
11.59; S, 10.42%.
Compound 18b: HPLC purity = 88% (C-18 NovaPak column;
MeOH:H2O/85:15), tr = 20 min; orange crystals from 1,4-dioxan
(78%), m.p. 172–174 °C; IR (KBr) cmꢁ1: 3548–3430, 3056, 2928,
2223, 1633, 1562; 1H-NMR (DMSO): d 0.83 (s, 3H), 1.31–2.83 (m,
5H), 2.82–2.97 (m, 4H), 3.11 (s, 3H), 3.33 (s, 1H), 6.42 (d, 1H,
J = 2.9 Hz), 6.61 (dd, 1H, J = 7.5 Hz and J = 2.9 Hz), 7.03–7.38 (m,
7H), 8.24 (s, 1H, D2O exchangeable), 9.06 (s, 1H, D2O exchange-
able); 13C-NMR (DMSO): d 13.9, 19.9, 21.5, 22.8, 23.1, 27.8, 31.8,
38.6, 40.9, 44.6, 50.8, 116.8, 120.3, 122.6, 124.8, 126.6, 130.9,
135.2, 137.4, 140.8, 142.3, 143.3, 148.8, 152.8, 162.8, 166.0. MS:
m/e = 468 (M+, 28%); Analysis Calcd for C28H28N4OS: C, 71.76; H,
6.02; N, 11.96; S, 6.84%. Found: C, 71.63; H, 5.84; N, 12.24; S, 6.77%.
Compound 18c: HPLC purity = 74% (C-18 NovaPak column;
MeOH:H2O/90:10), tr = 19 min; orange crystals from ethanol
(80%), m.p. 145–148 °C; IR (KBr) cmꢁ1: 3555–3426, 3054, 2931,
2221, 1635, 1560; 1H-NMR (DMSO): d 0.84 (s, 3H), 1.30–1.61 (m,
5H), 1.60 (dt, J = 7.2, 3.1 Hz, 3H), 1.73–1.88 (m, 2H), 2.44–2.68
(m, 2H), 2.90–2.95 (m, 2H), 6.43 (d, 1H, J = 3.4 Hz), 6.59 (dd, 1H,
J = 7.4 Hz and J = 3.4 Hz), 7.28–7.39 (m, 4H), 8.24 (s, 1H, D2O
exchangeable), 9.03 (s, 1H, D2O exchangeable); 13C-NMR (DMSO):
d 13.5, 19.9, 21.7, 23.5, 27.9, 31.8, 38.6, 41.6, 44.6, 50.6, 116.9,
120.6, 122.8, 124.3, 128.8, 130.8, 132.6, 134.8, 138.8, 140.6,
145.6, 148.8, 153.5, 162.4, 166.2. MS: m/e = 489 (M+, 31%); Analysis
Calcd for C27H25ClN4OS: C, 66.31; H, 5.15; Cl, 7.25; N, 11.46; S,
6.56%. Found: C, 66.28; H, 5.30; Cl, 7.40; N, 11.59; S, 6.70%.
Compound 18d: HPLC purity = 79% (C-18 NovaPak column;
MeOH:H2O/90:10), tr = 23 min; orange crystals from ethanol
(83%), m.p. 250–253 °C; IR (KBr) cmꢁ1: 3512–3427, 3053, 2929,
2223, 1635, 1562; 1H-NMR (DMSO): d 0.84 (s, 3H), 1.30–1.60 (m,
5H), 1.62 (dt, J = 7.3, 3.2 Hz, 3H), 1.73–1.89 (m, 2H), 2.84–2.89
(m, 2H), 2.92–2.99 (m, 2H), 3.01 (s, 3H), 6.40 (d, 1H, J = 3.2 Hz),
6.63 (dd, 1H, J = 7.9 Hz and J = 3.2 Hz), 7.28–7.42 (m, 4H), 8.24 (s,
1H, D2O exchangeable), 9.04 (s, 1H); 13C-NMR (DMSO): d 14.1,
19.6, 21.7, 23.3, 27.4, 31.9, 38.6, 41.2, 44.6, 51.4, 54.8, 117.3,
120.6, 123.1, 124.6, 125.4, 129.4, 135.2, 140.8, 142.6, 143.3,
149.5, 153.5, 154.9, 163.6, 166.0. MS: m/e = 484 (M+, 35%); Analysis
Calcd for C28H28N4O2S: C, 69.40; H, 5.82; N, 11.56; S, 6.62%. Found:
C, 69.53; H, 5.65; N, 11.71; S, 6.87%.
2.1.10. 6-((6bS,8aS,12aS,12bR)-4-hydroxy-8a-methyl-2,6b,7,8,8a,12,
12a,12b-octahydro-1H-naphtho[20,10:4,5]indeno[1,2-d]thiazol-10-yl)-
5-imino-2,4-diphenyl-4,5-dihydro-1,2,4-triazine-3(2H)-thione (19a),
6-((6bS,8aS,12aS,12bR)-4-hydroxy-8a-methyl-2,6b,7,8,8a,12,12a,
12b-octahydro-1H-naphtho [20,10:4,5] indeno[1,2-d]thiazol-10-yl)-5-
imino-4-phenyl-2-(p-tolyl)-4,5-dihydro-1,2,4-triazine-3(2H)-thione
(19b), 6-((6bS,8aS,12aS,12bR)-4-hydroxy-8a-methyl-2,6b,7,8,8a,12,
12a,12b-octahydro-1H-naphtho [20,10:4,5] indeno[1,2-d]thiazol-10-
yl)-5-imino-4-phenyl-2-(p-chlorophenyl)-4,5-dihydro-1,2,4-triazine-
3(2H)-thione (19c), 6-((6bS,8aS,12aS,12bR)-4-hydroxy-8a-methyl-2,
6b,7,8,8a,12,12a,12b-octahydro-1H-naphtho [20,10:4,5] indeno[1,2-d]
thiazol-10-yl)-5-imino-4-phenyl-2-(p-methoxyphenyl)-4,5-dihydro-
1,2,4-triazine-3(2H)-thione (19d)
General procedure: To a solution of either compound 18a
(0.454 g, 1 mmol), 18b (0.468 g, 1 mmol), 18c (0.489 g, 1 mmol)
or 18d (0.484 g, 1 mmol) in 1,4-dioxan (30 mL) containing triethyl-
amine (0.250 mL) phenylisothiocyanate (0.130 g, 1 mmol) was
added. The whole reaction mixture, in each case was heated under
reflux for 4 h then poured onto ice/water containing few drops of
hydrochloric acid and the formed solid product was collected by
filtration.
3. Results and discussion
3.1. Chemistry
Although much attention has been directed to study the uses of
estrone in heterocyclic synthesis [22–25], no investigations have
appeared in the literature to describe its uses in one-pot investiga-
tion to form pyran and pyrimidine ring systems. Therefore, the
need to create novel estrone derivatives for emerging drug targets
is an active area of medicinal chemistry. Recently, our research
group was involved through a series of heterocyclization of
Compound 19a: HPLC purity = 90% (C-18 NovaPak column;
MeOH:H2O/90:10), tr = 21 min; orange crystals from ethanol
(72%), m.p. 210–212 °C; IR (KBr) cmꢁ1: 3531–3424, 3054, 2925,

R.M. Mohareb et al. / Steroids 84 (2014) 46–56
51
pregnenolone to form thiophene, pyrazole and pyridine derivatives
as antitumor agents [26,27]. In the present work, we studied the
uses of estrone in heterocyclic chemistry through the one pot reac-
tion to form novel E-heterocyclic rings of estrone. The obtained
products were good candidates, in a different strategy, were used
to obtain new heterocyclic derivatives of estrone for comparison
with their cytotoxic activities towards human cancer and normal
cell lines. Thus, the one pot reaction of estrone (1) with either of
benzaldehyde (2a), 4-chlorobenzaldehyde (2b) or 4-methoxybenz-
aldehyde (2c) and either malononitrile (3a) or ethyl cyanoacetate
(3b) in ethanol containing triethylamine gave the pyran derivatives
4a–f, respectively (Scheme 1). The analytical and spectral data of
the synthesized products were based on their analytical and spec-
tral data. Thus, the 1H NMR spectrum of 4a (as an example)
showed beside the expected signals for estrone, a singlet at d
4.44 ppm (D2O exchangeable) corresponding to the NH2 group, a
singlet at d 5.77 ppm indicating the pyran H-4, a multiplet at d
6.88–7.36 ppm indicating the phenyl protons. Moreover, the 13C
NMR spectrum showed d 117.8 corresponding to the CN group
and signals at d 120.3, 124.2, 126.3, 128.6, 129.0, 135.4, 136.9,
140.3, 142.6, 146.8, 148.5, 149.5, 154.3, 155.9, 160.3 and 196.6
for the pyran and phenyl C. Formation of the pyran derivatives
using one pot reaction of cycloketone and cyanomethylene re-
agents were reported in literature [28,29].
Next, our program moved towards studying the reaction of
estrone with the aromatic aldehydes 2a–c and thiourea in etha-
nol containing triethyl amine gave the pyrimidine derivatives
6a–c, respectively. The 1H-NMR and 13C-NMR data of 6a–c were
the basis of their structure elucidation. Thus, the 1H NMR
spectrum of compound 6a (as an example) showed a singlet at
d 1.03 ppm indicating the SH group, a singlet at d 5.73 for the
pyrimidine H-4, a multiplet at d 6.93–7.42 ppm for the phenyl
protons a singlet at d 8.22 (D2O exchangeable) corresponding
to the presence of the NH group. Moreover, the 13C NMR
spectrum showed d 112.6, 113.8, 121.9, 124.8, 126.4, 127.2,
129.5, 135.4, 146.8, 153.0 154.8, 162.9 for the pyrimidine C-4
and aromatic carbons.
Compounds 6a–c with their high yields encouraged us to use
them as good candidates for fused thiazolopyrimidine formation.
Thus, the reaction of either of compounds 6a, 6b or 6c with any
of the
a-haloketones namely 2-bromo-1-phenylethanone (7a), 2-
bromo-1-(4-chlorophenyl)ethanone (7b), 2-bromo-1-p-tolyletha-
none (7c) and 2-bromo-1-(4-methoxy)ethanone (7d) in ethanol
solution under reflux gave the 3,7-diphenyl-7H-thiazolo[3,2-a]
O
H
H2C
X
CN
Ar-CHO
+
+
2a, Ar = C6H5
b, Ar = 4-Cl-C6H4
H
H
3a, X = CN
b, X = COOC2H5
c, Ar = 4-CH3O-C6H4
HO
1
EtOH/Et3N
X
O
CN
H
Ar
H
H
HO
Ar
X
4
a
C6H5
C6H5
NH2
OH
b
c
d
e
f
4-Cl-C6H4
4-Cl-C6H4
NH2
OH
NH2
4-CH3O-C6H4
4-CH3O-C6H4
OH
Scheme 1. Synthesis of compounds 4a-f.

52
R.M. Mohareb et al. / Steroids 84 (2014) 46–56
pyrimidine derivatives 8a–d, respectively (Scheme 2). Formation of
these thiazole derivatives took place in analogy to the well known
the Hantzsch thiazole synthesis which involves the condensation
to give the a-halocarbonyl compound 13 together with other
unidentified products. The pure 16-bromoestrone derivative was
separated via column chromatography and identified via the ana-
lytical and spectral data. Compound 13 reacted with thiourea in
ethanol gave the 2-aminothiazole derivative 14 (Scheme 3). The
structure of compound 14 was established on the basis of analyti-
cal and spectral data. Thus, the 1H NMR spectrum showed a singlet
of
a-haloketones with thioureas [30–33]. The obtained analytical
and spectral data of compounds 8a–d were in agreement with
their respective structures (see Section 2).
The reaction of estrone with phenylisothiocyanate and elemen-
tal sulphur in ethanol and triethyl amine gave the thiazole deriva-
tive 10. The reaction of compound 10 with either hydrazine
hydrate or phenylhydrazine gave the hydrazone derivatives 12a
and 12b, respectively.
at d 4.28 (D2O exchangeable) indicating the NH2 group, and the 13
C
NMR spectrum showed d at 120.3, 125.8, 128.6, 130.5, 134.7, 140.3,
154.7, 156.6 and 160.4 corresponding to the thiazole and phenyl
carbons.
The reaction of estrone with bromine has been reported before
under different reaction conditions. Thus, its reaction with N-bro-
mosuccinimide and refluxing chloroform [34,35] the product was
the 2,4-dibromo derivative which means that the reaction
occurred at phenolic ring A of estrone. On the other hand, in other
reports carrying the reaction of estrone acetate with bromine in
chloroform solution gave the 16-bromo-estrone acetate [36]. In
the present work direct bromination of estrone was carried out
to give the corresponding 16-bromo derivative by the shortest
route. Thus, estrone reacted with bromine in acetic acid at 60 °C
Similarly, the reaction of compound 13 with 2-cyanoethan-
ethioamide (15) in ethanol solution gave the thiazole derivative
15. The cyanomethylene moiety present in compound 15 showed
interesting reactivity. Thus, compound 15 reacted with the aryldi-
azonium salts 17a–d in ethanol containing sodium acetate to give
the arylhydrazone derivatives 18a–d, respectively. The reaction of
the latter compounds with phenylisothiocyanate (9) in ethanol
containing triethylamine gave the 1,2,4-triazin-6-thiazolyl deriva-
tives 19a–d, respectively (Scheme 4). The analytical and spectral
data of the latter products are consistent with their respective
2
2
6
5
6
4
3
6 4
3
6
5
6
4
3
6 4
2
6
5
6
4
3
6 4
3
6 4
6
5
6
4
3
6 4
3
6 4
Scheme 2. Synthesis of compounds 6a-c and 8a-d.

R.M. Mohareb et al. / Steroids 84 (2014) 46–56
53
Ph
N
S
EtOH/Et3N
PhNCS
S
1
+
+
S8
H
9
H
H
HO
10
Ph
N
NNHR
S
H
+
R-NHNH2
10
11a, R = H
b, R = Ph
H
H
HO
12a, R = H
b, R = Ph
O
Br
H
Br2/AcOH
1
+
Br2
H
H
HO
13
NH2
N
S
S
H
EtOH
13
+
H2N
NH2
5
H
H
HO
14
Scheme 3. Synthesis of 10, 12a,b; 13 and 14.
structures. Thus, the 1H NMR spectrum of 19a showed the
presence of a multiplet at d 7.26–7.44 ppm for the phenyl protons
and a singlet at d 8.22 (D2O exchangeable) for the NH group.
Moreover, the 13C NMR spectrum showed d 119.3, 121.4, 122.6,
124.3, 125.7, 126.4, 128.0, 129.8, 130.8, 133.2, 135.0, 140.8,
142.6, 143.4, 149.2, 152.8, 163.3, 166.0 corresponding to phenyl
carbons, d 168.3 corresponding to the exocyclic C@N group and
d 179.3 indicating the C@S group. It is clear from the demon-
strated reactions especially formation of 4a–f, 6a–c, 10 and 13 oc-
curred at the ring D of estrone and this is explained in terms of
the reactivity of the cyclopentanone moiety towards the respec-
tive reagents used for each reaction. More support for such
findings were demonstrated through the previous reported work
[10,37–39].
3.2. In vitro cytotoxic assay
3.2.1. Chemicals
Fetal bovine serum (FBS) and L-glutamine, were purchased from
Gibco Invitrogen Co. (Scotland, UK). RPMI-1640 medium was pur-
chased from Cambrex (New Jersey, USA). Dimethyl sulfoxide
(DMSO), doxorubicin, penicillin, streptomycin and sulforhodamine
B (SRB) were purchased from Sigma Chemical Co. (Saint Louis,
USA).
3.2.2. Cell cultures
Was obtained from the European Collection of cell Cultures
(ECACC, Salisbury, UK) and human gastric cancer (NUGC and HR),
human colon cancer (DLD1), human liver cancer (HA22T and

54
R.M. Mohareb et al. / Steroids 84 (2014) 46–56
CH2CN
N
S
EtOH
H
H2C
CN
CSNH2
+
13
H
H
15
HO
16
ArHN
N
C
N
CN
S
+
-
H
EtOH/NaOH
+
Ar-N
NCl
16
0-5 oC
17a, Ar =C6H5
H
H
b, Ar = 4-CH3-C6H4
c, Ar = 4-Cl-C6H4
HO
d, Ar = 4-CH3O-C6H4
18a, Ar =C6H5
b, Ar = 4-CH3-C6H4
c, Ar = 4-Cl-C6H4
d, Ar = 4-CH3O-C6H4
PhNCS/Et3N
9
Ar
N
S
N
N
Ph
N
NH
S
H
H
H
HO
19a, Ar =C6H5
b, Ar = 4-CH3-C6H4
c, Ar = 4-Cl-C6H4
d, Ar = 4-CH3O-C6H4
Scheme 4. Synthesis of compounds 16; 18a-d and 19a-d.
HEPG2), human breast cancer (MCF), nasopharyngeal carcinoma
(HONE1) and normal fibroblast cells (WI38) were kindly provided
by the National Cancer Institute (NCI, Cairo, Egypt). They grow as
monolayer and routinely maintained in RPMI-1640 medium sup-
plemented with 5% heat inactivated FBS, 2 mM glutamine and anti-
biotics (penicillin 100 U/mL, streptomycin 100 lg/mL), at 37 °C in a
humidified atmosphere containing 5% CO2. Exponentially growing
cells were obtained by plating 1.5 ꢀ 105 cells/mL for the seven hu-
man cancer cell lines including cells derived from 0.75 ꢀ 104 cells/
mL followed by 24 h of incubation. The effect of the vehicle solvent
(DMSO) on the growth of these cell lines was evaluated in all the
experiments by exposing untreated control cells to the maximum
concentration (0.5%) of DMSO used in each assay.
(DLD1), human liver cancer (HA22T and HEPG2), human breast
cancer (MCF), nasopharyngeal carcinoma (HONE1) and a normal
fibroblast cells (WI38). In this study the optimal cytotoxicity ef-
fects of the newly synthesized products were obtained using such
receptors. Moreover, in this study the receptor content being of the
same content among all cell lines. All of IC50 values were listed in
Table 1. Some heterocyclic compounds were observed with signif-
icant cytotoxicity against most of the cancer cell lines tested
(IC50 = 10–1000 nM). Normal fibroblasts cells (WI38) were affected
to a much lesser extent (IC50 > 10,000 nM). The reference com-
pound used is the CHS-828 which is a pyridyl cyanoguanidine
anti-tumor agent.
The heterocyclic estrone derivatives, prepared in this study,
were evaluated according to standard protocols for their in vitro
cytotoxicity against six human cancer cell lines including cells
derived from human gastric cancer (NUGC), human colon cancer
3.2.3. Structure activity relationship
From Table 1 it is clear that the estrone moiety was found to be
crucial for the cytotoxic effect of cyclic compounds 4a–f–19a–d.
Compounds 4c, 4f, 6b, 8b, 8c, 10, 13, 16, 18c and 19c exhibited

R.M. Mohareb et al. / Steroids 84 (2014) 46–56
55
Table 1
16 has higher cytotoxicity against the cell line DLDI than CHS
828. Next, for the arylhydrazo derivatives 18a–d, it is clear that
the substituted aryl derivatives 18b, 18c and 18d are more potent
than the unsubstituted aryl derivative 18a. Moreover, compound
18c showed the highest cytotoxicity against NUGC, DLDI and
MCF cell lines. The 1,2,4-triazine derivative 19c showed the highest
cytotoxicity among 19a–d, such high cytotoxicity is attributed to
the presence the 4-Cl-phenyl group. Our results showed that the
electronegative CN and Cl hydrophobic groups in the thiophene
derivative might play a very important role in enhancing the cyto-
toxic effect. The decahydronaphtho [20,10:4,5] indeno[1,2-b]pyran
derivative 4c selectively exhibited the maximum potency cytotoxic
activity against human gastric cancer NUGC, human colon cancer
cell lines (DLD1), and the human liver cancer HEPG2 with IC50’s
of 26, 53, 58 nM and 22 nM, respectively.
Cytotoxicity of novel estrone derivatives against a variety of cancer cell linesa [IC50
(nM)].
b
Compd
Cytotoxicity (IC50 in nM)
NUGC
DLDI
HA22T
HEPG2
HONE1
MCF
WI38
4a
4b
4c
4d
4e
4f
6a
6b
6c
8a
8b
8c
8d
10
12a
12b
13
822
455
26
214
2253
120
2355
378
53
120
2165
60
2549
42
1284
1620
46
129
255
1266
1283
960
38
2055
150
2560
55
2210
1920
58
1273
1988
85
3477
32
1577
1286
328
2655
288
22
1642
180
380
1266
1340
1319
2880
1246
1880
386
1880
2885
na
1655
1088
38
1320
180
1422
827
1288
633
664
1117
2866
3277
na
1640
39
2876
264
28
1189
2297
3283
4920
695
na
na
na
na
na
na
na
na
na
na
na
na
na
na
na
na
na
na
na
na
na
na
na
na
na
na
na
na
309
1438
1233
2193
304
287
1896
30
3472
160
227
2130
138
488
480
2810
3277
1154
38
1680
140
2279
660
48
884
485
2865
1286
4822
450
166
2416
188
2188
2788
70
1722
2877
1922
320
350
1886
15
1820
3228
1089
46
684
468
3163
780
166
1276
2275
774
29
1260
138
2788
120
255
4. Conclusions
In summary, we have shown herein that our strategy is compat-
ible with the synthesis of a wide range of steroids and particularly
steroids possessing a puran, thiazole, 1,2,4-triazine and some of
their derivatives as in a biologically important position C-17 of es-
trone skeleton. The cytotoxicity of the newly synthesized products
were evaluated against human gastric cancer (NUGC and HR), hu-
man colon cancer (DLD1), human liver cancer (HA22T and HEPG2),
human breast cancer (MCF), nasopharyngeal carcinoma (HONE1)
and normal fibroblast cells (WI38). The results showed that com-
pounds 4c, 4f, 6b, 8b, 8c, 10, 13, 16, 18c and 19c exhibited optimal
cytotoxic effect against cancer cell lines, with IC50’s in the nM
range.
14
16
18a
18b
18c
18d
19a
19b
19c
19d
CHS 828
122
489
3344
1666
130
44
548
894
664
2363
3280
180
620
2315
3220
2298
42
829
1466
2067
3866
2784
1829
2729
3276
1245
2781
25
18
a
NUGC, gastric cancer, DLDI, colon cancer, HA22T, liver cancer, HEPG2, liver
cancer; HONEI, nasopharyngeal carcinoma; HR, gastric cancer; MCF, breast cancer;
WI38, normal fibroblast cells.
CHS-828 is a pyridyl cyanoguanidine anti-tumor agent.
b
References
The sample concentration produces a 50% reduction in cell growth.
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optimal cytotoxic effect against cancer cell lines, with IC50’s in the
nM range. Comparing the cytotoxicity of the decahydronaphtho
[20,10:4,5] indeno[1,2-b]pyran derivatives 4a–f, it is obvious that
the cytotoxicity of 4c, 4d and 4f are higher than those of 4a, 4b
and 4e. The presence of the chloro group and either the NH2 or
the OH groups are responsible for the high potency of 4c and 4d.
The presence of the CH3O and the OH groups are responsible for
the reactivity of 4f, such high cytotoxicity is remarkable against
MCF cell line. On the other hand, considering the decahydro-naph-
tho [20,10:4,5] indeno[1,2-d]pyrimidine derivatives 6a–c, it is clear
that the un-substituted 4-phenyl group in 6b showed the less
potency. Compounds 6b with the 4-chlorophenyl moiety showed
more cytotoxicity than compound 6c with the 4-CH3O-phenyl moi-
ety. The decahydronaphtho [20,10:4,5] indeno[2,1-e]thiazolo[3,2-
a]pyrimidine derivatives 8a–d showed higher cytotoxicity than
the octahydro-1H-naphtho [20,10:4,5] indeno[1,2-d]thiazole
derivative 10. Such high cytotoxicity of 8a-d is attributed to the
thiazolo[3,2-a]pyrimidine moiety. For the 2-hydrazonothiazole
derivatives 12a and 12b, it is clear that the phenylhydrazono deriv-
ative 12b is more cytotoxic than the hydrazone derivative 12a
especially against the four cancer cell lines DLDI, HA22T, HEPG2
with IC50’s values 960, 46, 29 and 166, respectively. The bromo
derivative 13 showed high cytotoxicity against the cancer cell lines
NUGC and DLDI and such cytotoxicity effects are the highest
among the tested compounds. It is obvious that compounds 4c,
4f, 6b and 12b showed high cytotoxicity against the cancer cell
lines DLDI, HA22T and HEPG2 and that they are more potent than
the reference CHS 828. The 2-aminothiazole derivative 14 showed
low cytotoxicity against NUGC,DLDI cell lines and high cytotoxicity
against HA22T, HEPG2 and HONE1. On the other hand, the octahy-
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