Synthesis, characterization and biological evaluation of some 16β-azolyl-3β-amino-5α-androstane derivatives as potential anticancer agents

Article abstract of DOI:10.1016/j.ejmech.2011.05.030

A series of new 16β-azolyl-3β-amino-5α-androstane derivatives were synthesized and characterized. The new compounds were screened for their anticancer activity against the human cancer cell lines SW480, A549, HepG2, HeLa and SiHa in vitro using the MTT assay. The results of the in vitro study showed that a number of compounds have shown IC₅₀ values lower than 20 μM against the five cancer cell lines.

Full text of DOI:10.1016/j.ejmech.2011.05.030

European Journal of Medicinal Chemistry 46 (2011) 3662e3674
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis, characterization and biological evaluation of some 16
b-azolyl-3b-
amino-5 -androstane derivatives as potential anticancer agents
a
Hao Guo a, Guolan Zhang a, Tao Zhang a, Xianran He a, Zhongyuan Wu a, Yuling Xiao a, Yuanhu Pan b,
,
Guofu Qiu a, Peng Liu a, Xianming Hu a
*
a State Key Laboratory of Virology, Ministry of Education Laboratory of Combinatorial Biosynthesis and Drug Discovery, Wuhan University School of Pharmaceutical Science,
Wuhan 430071, China
b MOA Key Laboratory of Food Safety Evaluation/National Reference Laboratory of Veterinary Drug Residues, Huazhong Agricultural University, Wuhan 430070, China
a r t i c l e i n f o
a b s t r a c t
Article history:
A series of new 16b-azolyl-3b-amino-5a-androstane derivatives were synthesized and characterized. The
new compounds were screened for their anticancer activity against the human cancer cell lines SW480,
A549, HepG2, HeLa and SiHa in vitro using the MTT assay. The results of the in vitro study showed that
Received 14 March 2011
Received in revised form
10 May 2011
Accepted 14 May 2011
Available online 23 May 2011
a number of compounds have shown IC50 values lower than 20 mM against the five cancer cell lines.
Ó 2011 Elsevier Masson SAS. All rights reserved.
Keywords:
Androstane
Azoles
Heterocycles
Amine
Synthesis
Anticancer activity
1. Introduction
Nitrogen containing steroids are an important type of steroids,
including natural products, and semi-synthetic compounds and
synthetic compounds, all of which have been studied intensively.
Most modifications are aimed at the A- and D-rings by incorpora-
tion of amino groups to the steroids backbone [10e15].
Nitrogen containing steroids have the ability to regulate
a variety of biological processes and thus are potential drug
candidates for the treatment of a large number of diseases
including breast cancer [16], prostate cancer [17,18], leukemia
[19e21], autoimmune diseases [22], and osteoporosis [23]. A series
of 16-, 20-, and 21-aminosteroids have been synthesized, and these
novel class of compounds have shown excellent anticancer or
antioxidant activities [24e26]. It has been reported that some
According to the World Health Organization (WHO), cancer is
presently the second most important disease leading to death in
both developing and developed countries. Drugs that are currently
available for the treatment of cancer include bioalkylating agents
(chlormethine) [1,2], antimetabolic agents (fluorouracil) [3,4],
anticancer antibiotics (doxorubicin) [5], and compounds from
plants and their derivatives (paclitaxel) [6]. However, sufficient
doses of anticancer drugs to kill tumor cells are often toxic to
normal tissues, thus leading to numerous side effects, which, in
turn, limits treatment efficacy. Long term effectiveness is often
limited by dose-related cumulative cardiotoxicity and development
of acquired drug resistance [7e9]. Therefore, development of new
therapeutic agents for the treatment of cancer that are more active
and less side effects producing is of utmost importance.
tertiary aminosteroids, such as 2
b, 16b-dipiperidino-5a-andros-
tane-3 , 17 -diol dipivalate(DAP) (Fig. 1) [27] exhibit significant
a
b
antitumor activity against transplantable tumors in three animal
species, including mice, hamsters, and rats. In an effort to find the
new analogs of this type of novel steroid molecule, some 16E-ary-
lidene androstene derivatives have emerged as potent anticancer
agents, such as compound DPJ-RG-1110 (Fig. 1). Further, a struc-
tureeactivity relationship study has indicated that incorporation of
a heterocyclic ring into the steroid backbone at position C-3 and
Steroids elicit diverse biological actions via various functional
groups located around the periphery of their rigid tetracyclic core.
* Corresponding author. Tel.: þ86 27 68759887; fax: þ86 27 68759850.
E-mail address: cassie_gh@hotmail.com (X. Hu).
0223-5234/$ e see front matter Ó 2011 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2011.05.030

H. Guo et al. / European Journal of Medicinal Chemistry 46 (2011) 3662e3674
3663
Fig. 1. Structures of DAP, DPJ-RG-1110 and CYP17 inhibitors.
variation of the substituent groups at position C-16 bring about
important impacts to the activity of a steroid backbone [28e30].
Azoles, a versatile group of heterocycles often show some
special biological activities when they are introduced to biologically
active compounds. Some 1,2,4-triazole, 1,2,3-triazole, 1,3,4-
thiadiazole, benzimidazole and benzotriazole derivatives have
been reported to possess antibacterial [31,32], antivirus [33], anti-
microbial [34], anti-inflammatory [35,36], antiglaucoma [37],
antiprotozoal [38,39] and anticancer activities [40,41]. Moreover,
when azole groups were introduced to the steroids backbone, some
resulting C-21 and C-17 azolylsteroids were found to be potent
The chemical structures of the compounds synthesized were
elucidated on the basis of 1H NMR, 13C NMR, MS and elemental
analysis. The detailed physical and analytical data are given in
experimental part.
3. Results and discussion
3.1. Chemistry
X-ray crystal structure of 16f and 18d were obtained and are
presented in Figs. 2 and 3 respectively. These structures confirm the
inhibitors of 17a-hydroxylase-C17,20-lyase(CYP17 inhibitors) (Fig. 1)
b-stereochemistry of the heterocycles at C-3, the azoles at C-16 and
which is the key enzyme for androgen biosynthesis [42e46].
Inhibitors of such enzymes are useful for the treatment of androgen
dependant prostate cancer. Much work has been done on C-21 and
C-17 azolylsteroids in searching for new potential CYP17 inhibitors
[47e49]. An increasing numbers of publications had recently
reported the anticancer activities of a wide range of natural [50,51],
semi-synthetic [52,53] and synthetic aminosteroidal compounds
[54,55]. However, to the best of our knowledge, little attention has
been given to 16-azolyl-3-amino-androstanes.
the acetoxy group at C-17. Crystallographic data for 16f and 18d
have been deposited with the Cambridge Crystallographic Data
Centre with the deposition number CCDC 805534 and CCDC
805535. These data can be obtained free of charge on application to
CCDC, 12 Union Road, Cambridge CB2 1EZ, UK [fax(þ44) 1223
336033, e-mail: deposit@ccdc.cam.ac.uk].
3.2. Anticancer activity
On the basis of the above details, we synthesized a series of
androstane-based aminosteroids to screen their anticancer activity
against different cancer cell lines in vitro in order to get more
potent anticancer agents.
The newly synthesized compounds 16aeg, 17aeg, 18aeg, 19aeg
were evaluated for their anticancer activities against human cancer
cell lines derived from various human cancer types, including HeLa
(human cervical cancer cells), SW480 (human colon adenocarci-
noma cells), A549 (human lung carcinoma cells), HepG2 (human
hepatic carcinoma cells) and SiHa (human cervical cancer cells).
293 (human embryonic cells) is non-cancer cell line. In vitro eval-
uation of the anticancer activities of the synthesized compounds
was carried out using the 3-(4,5-dimethylthiazol-2-yl)-2,5-
diphenyltetrazolium bromide (MTT) assay. Doxorubicin was
employed as the positive control. The anticancer potency of these
compounds was indicated by IC50 values that were calculated by
linear regression analysis of the concentrationeresponse curves
obtained for each compound. The results are summarized in
Table 1.
2. Chemistry
The routes utilized for the synthesis of our target compounds
16aeg, 17aeg, 18aeg, 19aeg are outlined in Scheme 1. 5
stan-16 -bromo-3 -ol-17-one 2 was prepared with epiandroster-
one and CuBr2. The adol product was then oxidized with Jones
reagent to obtain the compound 3 [56e58]. The 16 -azolyl deriv-
a-andro-
a
b
b
atives 4, 5, 6, 7 were prepared with appropriate azoles. Then the C-3
amino derivatives 8aeg, 9aeg, 10aeg, 11aeg were obtained via
reductive amination of the carbonyl group at the C-3 position using
pyrrolidine, piperidine, morpholine, N-methylpiperazine, diethyl-
amine, N-methylaniline and N-benzylmethylamine respectively.
Compounds 12aeg, 13aeg, 14aeg, 15aeg were prepared by
reduction with sodium borohydride in methanol. Next, the esteri-
fication of the 17b-ol derivatives with acetic anhydride and 4-
dimethylaminopyridine (DMAP) refluxed in CHCl2 provided the
desired ester 16aeg, 17aeg, 18aeg, 19aeg.
As shown in Table 1, some of the tested compounds showed
moderate to potent anticancer activities against all the tested cells.
Six of the twenty-eight new compounds showed strong anticancer
activities against more than three cell lines (IC50 < 20
Compounds 18f and 19g were the most promising compounds
amongst the tested derivatives, with IC50 values of 5.12e18.63
and 4.22e15.76 M, respectively.
mM).
mM
m

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H. Guo et al. / European Journal of Medicinal Chemistry 46 (2011) 3662e3674
Scheme 1. Preparation of target compounds (16aeg, 17aeg, 18aeg, 19aeg). Reagents and Conditions: (a) CuBr2, Methanol, 65ꢀC, 12h; (b) Jones Reagent, acetone, Icebath; (c)
appropriate azoles, K2CO3, CH3CN, 65-80ꢀC; (d) appropriate amine, NaBH3CN, Methanol, 40ꢀC, 24h; (e) NaBH4, Methanol, Icebath, 4h; (f) Acetic Anhydride, 4-Dimethylamino-
pyridine, CH2CI2, 40ꢀC.
Most of the 16
b
-triazole derivatives, including 16a, 16b, 17a and
better activities (16c for HeLa 22.32
for HepG2 23.52 M). When the nitrogenous heterocycles were
replaced by diethylamine, 16e and 17e exhibited no activity
(>50 M). This result suggested that an aliphatic amine at the C-3
mM, for HepG2 19.47 mM; 16d
17b which have pyrrolidine and piperidine groups, respectively, at
the C-3 position exhibited no activity against all the tested cells
m
(IC50 > 50
m
M). Compounds 16c and 17c, which have a morpholine
m
moiety at the C-3 position, and compounds 16d and 17d which
position was not beneficial for anticancer activity. On the other
have a methylpiperazine moiety at the C-3 position, have slightly
hand, the introduction of aromatic amines to the C-3 position
Fig. 2. X-ray crystal structure of 16f.

H. Guo et al. / European Journal of Medicinal Chemistry 46 (2011) 3662e3674
3665
Fig. 3. X-ray crystal structure of 18d.
resulted in increased anticancer activity. Compounds 16f and 17f
which have N-methylaniline moiety at the C-3 position and
compounds 16g and 17g, which have N-benzylmethylamine moiety
at the C-3 position, showed potent activity against most of the five
the most potent anticancer activities, with IC50 values ranging from
4.22 M to 15.76 M against most of the five cancer cell lines. These
m
m
findings indicated that incorporation of benzimidazole or benzo-
triazole to the steroid scaffolds at the C-16 position together with
the aromatic amine at C-3 position played an important role for the
activity. Furthermore, it appeared that introducing proper azoles at
the C-16 position could serve as a promising launch point for
further design of this type of steroidal anticancer agent. All the
active compounds did not show obvious cell line selectivity.
Compounds 18f, 18g, 19f and 19g were selected for further
in vitro evaluation using 293 cell line which is a non-cancer cell
line. The results are summarized in Table 2. As shown in Table 2,
among the four tested compounds, only compound 18g showed
cancer cell lines (16f for A549 12.34
SiHa 11.74 M; 16g for HepG2 10.56
HepG2 8.43 M).
A comparison of the results obtained from the 16
benzimidazole and 16 -benzotriazole derivatives, showed that
mM; 17f for HepG2 9.35
m
M, for
m
mM; 17g for A549 11.41
m
M,
m
b
-triazole, 16b-
b
compounds 18aeg and 19aeg which possess benzimidazole and
benzotriazole moieties displayed good activities. Compared with
16
18b and 19b were active against some cancer cell lines (18b for
A549 22.72 M; 19b for SiHa 18.74 M). Similar findings were
observed amongst the 3 -aromatic amine and the 3 -aliphatic
b-triazolyl-3b-heterocyclic derivatives 16b and 17b, compounds
m
m
little cytotoxicity (IC50 48.70
mM). Compounds 18f, 19f and 19g
b
b
exhibited no cytotoxicity (IC50 > 80 mM). It is apparent from the
amine derivatives. Among all the tested compounds, the N-meth-
ylaniline and N-benzylmethylamine analogs 19f and 19g exhibited
results depicted in Table 2 that compounds 18f, 19f and 19g exhibit
obvious selectivity between cancer cell line and non-cancer cell
line.
In an overall view, aromatic amines at the C-3 position and aryl
azoles at the C-16 position appeared to play a critical role for the
anticancer activity. General structureeactivity relationship of
compounds for anticancer activity in vitro based on the IC50 results
on the five cancer cells lines is presented schematically in Fig. 4.
Table 1
Anticancer activity data for compounds 16aeg, 17aeg, 18aeg, 19aeg against HeLa,
SW480, A549, HepG2, and SiHa.
a
Compounds
IC50/(
HeLa
m )
mol Lꢁ1
SW480
A549
HepG2
SiHa
4. Conclusion
16a
16b
16c
16d
16e
16f
16g
17a
17b
17c
17d
17e
17f
17g
18a
18b
18c
18d
18e
18f
18g
19a
19b
19c
19d
19e
19f
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
31.78
>50
>50
>50
>50
>50
>50
>50
18.85
>50
18.63
>50
>50
>50
>50
>50
>50
9.22
15.76
0.06
>50
>50
>50
39.70
>50
12.34
>50
>50
>50
40.29
>50
>50
18.86
11.41
>50
22.72
12.18
8.99
>50
5.12
7.72
39.65
>50
16.12
22.40
>50
28.43
4.22
0.36
>50
>50
19.47
23.52
>50
21.85
10.56
>50
>50
>50
>50
>50
9.35
8.43
>50
>50
>50
31.71
>50
6.36
5.54
>50
29.57
>50
11.71
22.11
>50
5.14
0.5
>50
>50
>50
>50
>50
37.13
22.43
>50
>50
47.81
>50
>50
11.74
15.17
>50
>50
>50
22.03
37.22
9.83
26.18
37.14
18.74
12.27
14.92
29.83
6.67
In summary, a series of new 16b-azolyl-3b-amino-5a-andros-
22.32
>50
>50
19.77
18.71
>50
>50
>50
38.12
>50
72.12
tane derivatives were synthesized and evaluated for their anti-
cancer activities. Some of the synthesized compounds exhibited
potent anticancer activities against the five tested cancer cell lines
in vitro. In particular, compound 18f and 19g were the most
promising derivatives, with IC50 values ranging from 5.12
mM to
18.63 M, 4.22 Me15.76 M, against all the five cancer cell lines
m
m
m
respectively. Aromatic amines at the C-3 position and aryl azoles at
the C-16 position are beneficial for the activity. Substituent changes
to the A- and D-rings of the androstane skeleton can affect potency
against different kinds of cancer cell lines. In an overall view,
>50
>50
29.62
19.82
>50
>50
10.41
11.33
>50
>50
>50
>50
>50
Table 2
Cytotoxic activity data for compounds 18f, 18g, 19f, 19g
against 293 cell line.
a
Compounds
IC50/(m )
M Lꢁ1
293
18f
18g
19f
19g
>80
48.70
>80
>80
4.51
6.04
0.23
19g
Doxorubicin
11.9
0.45
Exponentially growing 293 cells were treated with the
compounds at different concentrations for 48 h. Cell-
growth inhibition was analyzed by the MTT assay.
Exponentially growing cells were treated with the compounds at different
concentrations for 48 h. Cell-growth inhibition was analyzed by the MTT assay.
a
IC50 is the concentration of compound that inhibits 50%
a
IC50 is the concentration of compound that inhibits 50% of cell growth.
of cell growth.

3666
H. Guo et al. / European Journal of Medicinal Chemistry 46 (2011) 3662e3674
Fig. 4. General SAR of compounds for anticancer activity in vitro based on the IC50 results on the five cancer cells lines.
aromatic amines at the C-3 position and aryl azoles at the C-16
position appeared to play a critical role for the anticancer activity.
Preliminary structureeactivity relationships were put forward
based on the biological results. Future work will focus on the
synthesis of additional candidate structures to address specific
cancer cell lines.
5.1.2.1. 16a-Bromo-5a-androstan-3,17-dione (3). White solid. yield
97%. m.p. 170e171 ꢀC; 1H NMR(400 MHz,CDCl3):
d 0.93 (s, 3H, CH3-
19), 1.04 (s, 3H, CH3-18), 4.54 (d, 1H, J ¼ 3.32 Hz, H-16). 13C
NMR(150 MHz,CDCl3): d 6.89, 9.65, 16.05, 23.92, 25.84, 27.74, 29.54,
29.67, 31.26, 33.46, 33.77, 39.96, 41.65, 41.93, 43.24, 49.14, 206.80,
208.64. Anal. Calcd. for C19H27BrO2: C 62.13, H 7.41; Found C 62.32, H
7.52.
5. Experimental protocols
5.1.3. General procedure for the synthesis of compounds 4, 5, 6, 7
To a suspension of 16a-bromo-5a-androstan-3,17-dione 3 0.37 g
5.1. Chemistry
(1.00 mmol) in 20 ml of CH3CN was added 0.21g (3.00 mmol,
3.0 equiv) of 1,2,4-triazole and 0.27g (2.00 mmol, 2.0 equiv) of
K2CO3. The resulting solution was stirred for 24 h at 80 ꢀC until the
reaction was completed (monitored by TLC). The CH3CN was
removed under reduced pressure, 20 ml CH2Cl2 was added to the
residue, the organic layer was washed with saturated NaHCO3,
brine and dried over anhydrous Na2SO4, filtered. The filtrate was
concentrated in reduced pressure to afford a crude product. The
products were purified by flash column chromatography to yield
compound 4. Compounds 5, 6 and 7 were obtained with 1,2,3-
triazole, 1H-benzotriazole and benzimidazole respectively by
using the same method that described above.
All the chemicals and solvent were purchased from commercial
sources. Solvents and reagents were dried and purified according to
the literature methods. Melting points were determined on a XT-4
apparatus (uncorrected). 1H NMR and 13C NMR spectra were
obtained on a Varian Mercury VX400 apparatus in CDCl3 with TMS
as internal standard. Mass spectra were recorded on a Shimadzu
LCMS-2020 spectrometer. X-ray diffraction data were collected on
Bruker SMART APEX CCD diffractometer. Elemental analysis was
carried out on a VarioEL III (German) instrument. Silica gel was
used for analytical and flash chromatography.
5.1.1. General procedure for the synthesis of compounds 2
5.1.3.1. 16
White solid. yield 60%. m.p. 157e159 ꢀC; 1H NMR(400 MHz,CDCl3):
1.00 (s, 3H, CH3-19),1.08 (s, 3H, CH3-18), 4.52 (t,1H, C16 H), 7.88 (s,
1H), 8.10 (s,1H).13C NMR(150MHz,CDCl3):
11.43,14.20, 20.48, 28.40,
29.26, 30.63, 31.79, 34.05, 35.83, 37.97, 38.28, 44.44, 46.46, 46.89,
47.37, 53.85, 63.86, 144.03, 152.13, 211.34, 211.70. Anal. Calcd. for
C21H29N3O2: C 70.95, H 8.22, N 11.82; Found C 70.80, H 8.34, N 11.76.
b-(1H-1,2,4-triazol-1-yl)-5a-androstan-3,17-dione (4).
To a stirred solution of epiandrosterone (1.45 g, 5.00 mmol) in
methanol was added the CuBr2 (2.23g, 10.00 mmol, 2.0 equiv). The
resulting solution was stirred for 12 h at 65 ꢀC until the reaction was
completed (monitored by TLC). The reaction was quenched and
concentrated inreduced pressure. Then 20ml CH2Cl2 and 10 ml water
was added to the residue. The organic layer was washed with brine
and dried over anhydrous Na2SO4, filtered. The filtrate was concen-
trated in reduced pressure to afford a crude product. The products
were purified by flash column chromatography to yield compound 2.
d
-a
d
5.1.3.2. 16
White solid. yield 62%. m.p. 178e180 ꢀC; 1H NMR(400 MHz,CDCl3):
1.07 (s, 3H, CH3-19),1.15 (s, 3H, CH3-18), 4.59 (t,1H, C16 H), 7.95 (s,
1H), 8.17 (1H, s). 13C NMR(150 MHz,CDCl3):
11.41,14.32, 20.50,
b-(1H-1,2,3-triazol-1-yl)-5a-androstan-3,17-dione (5).
d
-a
5.1.1.1. 5
a
-Androstan-16
a-bromo-3
b
-ol-17-one (2). White solid.
yield 91%. m.p. 164e165 ꢀC; 1H NMR(400 MHz,CDCl3):
d
0.82 (s, 3H,
d
28.29, 29.25, 30.61, 31.80, 34.03, 35.82, 37.97, 38.18, 44.39, 46.44,
46.90, 47.37, 53.83, 63.85, 144.00, 152.10, 211.33, 211.72. Anal. Calcd.
for C21H29N3O2: C 70.95, H 8.22, N 11.82; Found C 70.89, H 8.41, N
11.70.
CH3-19), 0.89 (s, 3H, CH3-18), 3.59 (m, 1H, H-3), 4.52 (d, 1H,
J ¼ 6.34 Hz, H-16). 13C NMR(150 MHz,CDCl3):
d 12.26, 14.20, 20.39,
28.23, 30.72, 31.36, 32.33, 34.07, 34.29, 35.63, 36.84, 37.97, 44.47,
46.41, 47.82, 47.93, 54.23, 71.05, 213.60. Anal. Calcd. for C19H29BrO2:
C 61.79, H 7.91; Found C 61.58, H 7.88.
5.1.3.3. 16
White solid. yield 55%. m.p. 209e211 ꢀC; 1H NMR(400 MHz,CDCl3):
1.10 (s, 3H, CH3-19), 1.27 (s, 3H, CH3-18), 4.99 (t,1H, C16 H), 7.33 (t,
1H), 7.40e7.51 (m, 2H), 8.07 (d, 1H,
8.36Hz). 13C
NMR(150 MHz,CDCl3): 11.50, 14.68, 20.56, 28.48, 29.33, 30.76,
b-(1H-benzotriazol-1-yl)-5a-androstan-3,17-dione (6).
5.1.2. General procedure for the synthesis of compounds 3
d
-a
To a solution of aldol products (5.00 mmol) 2 in acetone, Jones
reagent was added drop wise. The mixture was stirred at 0 ꢀC until
the reaction was completed (monitored by TLC). The reaction was
quenched with ethanol (3 ml) and concentrated in reduced pres-
sure. The crude product was dissolved in CH2Cl2, the organic layer
was washed with brine and dried over anhydrous Na2SO4, filtered.
The filtrate was concentrated in reduced pressure to afford a crude
product. The products were purified by flash column chromatog-
raphy to yield compound 3. The Jones reagent was prepared
according to previous study [56e58].
J
¼
d
31.94, 34.24, 35.93, 38.04, 38.39, 44.51, 46.59, 47.39, 47.77, 54.03,
62.53, 109.42, 120.01, 124.15, 127.68, 133.98, 145.69, 211.43, 211.87.
Anal. Calcd. for C25H31N3O2: C 74.04, H 7.70, N 10.36; Found C 73.88,
H 7.61, N 10.40.
5.1.3.4. 16
b-(1H-benzimidazol-1-yl)-5a-androstan-3,17-dione (7).
White solid. yield 58%. m.p. 195e197 ꢀC; 1H NMR(400 MHz,CDCl3):

H. Guo et al. / European Journal of Medicinal Chemistry 46 (2011) 3662e3674
3667
d
1.07 (s, 3H, CH3-19), 1.12 (s, 3H, CH3-18), 4.74 (t, 1H, C16
-
a
H),
5.1.4.7. 16
androstan-17-one (8g). White solid. yield 51%. m.p. 169e172 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.86 (s, 3H, CH3-19), 1.12 (s, 3H, CH3-18),
2.20 (s, 3H, NeCH3), 3.57 (s, 2H, PhCH2), 4.57 (t, 1H, C16 H), 7.31 (s,
5H), 7.95 (s, 1H), 8.16 (s, 1H). Anal. Calcd. for C29H40N4O: C 75.61, H
8.75, N 12.16; Found C 75.51, H 8.68, N 12.31.
b-(1H-1,2,4-triazol-1-yl)-3b-(N-benzylmethylamino)-5a-
7.28e7.32 (m,2H), 7.39 (d, 2H, J ¼ 8.40Hz), 7.80 (s, 1H). 13C
NMR(150 MHz,CDCl3):
d
11.46, 14.64, 20.54, 28.39, 30.30, 31.74,
d
34.09, 35.89, 37.99, 38.29, 44.45, 46.50, 47.11, 47.70, 53.97, 60.71,
109.76, 120.44, 122.60, 123.27, 134.17, 140.58, 143.40, 211.37, 213.12.
Anal. Calcd. for C26H32N2O2: C 77.19, H 7.97, N 6.92; Found C 77.08, H
8.09, N 6.87.
-a
5.1.4.8. 16
one (9a). White solid. yield 51%. m.p.152e156 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.87 (s, 3H, CH3-19), 1.12 (s, 3H, CH3-18),
2.57 (s, 4H, NCH2), 4.57 (t,1H, C16 H), 7.93 (s, 1H), 8.14 (s, 1H). Anal.
Calcd. for C25H38N4O: C 73.13, H 9.33, N 13.65; Found C 73.22, H
9.40, N 13.51.
b-(1H-1,2,3-triazol-1-yl)-3b-pyrrolidino-5a-androstan-17-
5.1.4. General procedure for the synthesis of compounds 8aeg, 9aeg,
10aeg, 11aeg
d
To a suspension of dione 4 (1.00 mmol) in 20 ml of absolute
methanol was added 0.5 ml of pyrrolidine and 60 mg (1.20 mmol,
1.2 equiv) of NaBH3CN, then the solution was acidified to pHz6 with
acetic acid. The resulting mixture was stirred at 40 ꢀC for 24 h. The
methanol was removed under reduced pressure, 20 ml CH2Cl2 was
added to the residue, the organic layer was washed with saturated
NaHCO3, brine and dried over anhydrous Na2SO4, filtered. The filtrate
was concentrated in vacuum to afford a crude product. The product
was purified by flash column chromatography to yield compound 8a.
Compounds 8beg were obtained with piperidine, morpholine,
N-methylpiperazine, diethylamine, N-methylaniline and N-benzyl-
methylamine respectively. Compounds 9aeg, 10aeg, 11aeg,were
prepared by using the same method that described above.
-a
5.1.4.9. 16
one (9b). White solid. yield 62%. m.p.170e173 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.88 (s, 3H, CH3-19), 1.11 (s, 3H, CH3-18),
2.57 (s, 4H, NCH2), 4.57 (t,1H, C16 H), 7.93 (s, 1H), 8.14 (s, 1H). Anal.
b-(1H-1,2,3-triazol-1-yl)-3b-piperidino-5a-androstan-17-
d
-
a
Calcd. for C26H40N4O: C 73.54, H 9.49, N 13.19; Found C 73.36, H
9.58, N 13.31.
5.1.4.10. 16
17-one (9c). White solid. yield 55%. m.p.181e184 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.87 (s, 3H, CH3-19), 1.11 (s, 3H, CH3-18),
2.57 (s, 4H, NCH2), 3.75 (s, 4H, OCH2), 4.57 (t, 1H, C16 H), 7.94 (s,
b-(1H-1,2,3-triazol-1-yl)-3b-morpholino-5a-androstan-
d
5.1.4.1. 16
one (8a). White solid. yield 57%. m.p. 188e192 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.84 (s, 3H, CH3-19), 1.11 (s, 3H, CH3-18),
2.57 (s, 4H, NCH2), 4.57 (t, 1H, C16 H), 7.94 (s, 1H), 8.16 (s, 1H). Anal.
b
-(1H-1,2,4-triazol-1-yl)-3
b
-pyrrolidino-5
a
-androstan-17-
-a
1H), 8.14 (s, 1H). Anal. Calcd. for C25H38N4O2: C 70.39, H 8.98, N
13.13; Found C 70.50, H 8.87, N 13.10.
d
-a
Calcd. for C25H38N4O: C 73.13, H 9.33, N 13.65; Found C 73.02, H
9.18, N 13.55.
5.1.4.11. 16
androstan-17-one (9d). White solid. yield 65%. m.p.169e173 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.87 (s, 3H, CH3-19), 1.12 (s, 3H, CH3-18),
2.27(s, 3H, NeCH3), 2.40e2.56 (br, 8H, NCH2), 4.57 (t, 1H, C16 H),
7.94 (s, 1H), 8.16 (s,1H). Anal. Calcd. for C26H41N5O: C 71.03, H 9.40,
N 15.93; Found C 71.10, H 9.25, N 15.87.
b-(1H-1,2,3-triazol-1-yl)-3b-(4-Methylpiperazinyl)-5a-
d
5.1.4.2. 16
one (8b). White solid. yield 51%. m.p. 184e187 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.82 (s, 3H, CH3-19), 1.11 (s, 3H, CH3-18),
2.52 (s, 4H, NCH2), 4.57 (t,1H, C16 H), 7.94 (s, 1H), 8.15 (s,1H). Anal.
b
-(1H-1,2,4-triazol-1-yl)-3
b
-piperidino-5
a
-androstan-17-
-a
d
-a
Calcd. for C26H40N4O: C 73.54, H 9.49, N 13.19; Found C 73.36, H
9.58, N 13.03.
5.1.4.12. 16
17-one (9e). White solid. yield 61%. m.p.198e201 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.88 (s, 3H, CH3-19), 1.12 (s, 3H, CH3-18),
2.52 (s, 4H, NCH2), 4.57 (t,1H, C16 H), 7.94 (s, 1H), 8.14 (s, 1H). Anal.
Calcd. for C25H40N4O: C 72.77, H 9.77, N 13.58; Found C 72.60, H
9.63, N 13.68.
b-(1H-1,2,3-triazol-1-yl)-3b-diethylamino-5a-androstan-
d
5.1.4.3. 16
one (8c). White solid. yield 54%. m.p. 165e168 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.83 (s, 3H, CH3-19), 1.12 (s, 3H, CH3-18),
2.56 (s, 4H, NCH2), 3.72 (s, 4H, OCH2), 4.57 (t, 1H, C16 H), 7.94 (s,
b
-(1H-1,2,4-triazol-1-yl)-3b-morpholino-5a-androstan-17-
-a
d
-a
1H), 8.16 (s, 1H). Anal. Calcd. for C25H38N4O2: C 70.39, H 8.98, N
13.13; Found C 70.21, H 8.89, N 13.02.
5.1.4.13. 16
androstan-17-one (9f). White solid. yield 56%. m.p. 169e173 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.88 (s, 3H, CH3-19), 1.11 (s, 3H, CH3-18),
2.82 (s, 3H, NeCH3), 4.57 (t, 1H, C16 H), 6.69 (t, 1H), 6.79 (d, 2H,
b-(1H-1,2,3-triazol-1-yl)-3b-(N-methylphenylamino)-5a-
d
5.1.4.4. 16
rostan-17-one (8d). White solid. yield 54%. m.p. 179e181 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.86 (s, 3H, CH3-19), 1.11 (s, 3H, CH3-18),
2.27(s, 3H, NeCH3), 2.39e2.53 (br, 8H, NCH2), 4.57 (t, 1H, C16 H),
b
-(1H-1,2,4-triazol-1-yl)-3
b
-(4-Methylpiperazinyl)-5
a-and
-a
J ¼ 8.30Hz), 7.23(t, 2H), 7.96 (s, 1H), 8.16 (s, 1H). Anal. Calcd. for
d
C28H38N4O: C 75.30, H 8.58, N 12.54; Found C 75.22, H 8.63, N 12.39.
-a
7.94 (s,1H), 8.16 (s,1H). Anal. Calcd. for C26H41N5O: C 71.03, H 9.40, N
15.93; Found C 71.17, H 9.28, N 15.86.
5.1.4.14. 16
androstan-17-one (9g). White solid. yield 57%. m.p. 155e158 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.85 (s, 3H, CH3-19), 1.11 (s, 3H, CH3-18),
2.21 (s, 3H, NeCH3), 3.57 (s, 2H, PhCH2), 4.56 (t, 1H, C16 H), 7.31 (s,
5H), 7.95 (s, 1H), 8.16 (s, 1H). Anal. Calcd. for C29H40N4O: C 75.61, H
8.75, N 12.16; Found C 75.51, H 8.68, N 12.21.
b-(1H-1,2,3-triazol-1-yl)-3b-(N-benzylmethylamino)-5a-
d
5.1.4.5. 16
17-one (8e). White solid. yield 52%. m.p. 228e230 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.82 (s, 3H, CH3-19), 1.11 (s, 3H, CH3-18),
2.52 (s, 4H, NCH2), 4.57 (t,1H, C16 H), 7.94 (s, 1H), 8.15 (s,1H). Anal.
b
-(1H-1,2,4-triazol-1-yl)-3
b-diethylamino-5a-androstan-
-a
d
-a
Calcd. for C25H40N4O: C 72.77, H 9.77, N 13.58; Found C 72.61, H
9.66, N 13.70.
5.1.4.15. 16
androstan-17-one (10a). White solid. yield 56%. m.p. 182e185 ꢀC;
1H NMR(400 MHz,CDCl3):
0.87 (s, 3H, CH3-19), 1.24 (s, 3H, CH3-
18), 2.57 (s, 4H, NCH2), 4.98 (t, 1H, C16 H), 7.37 (t, 1H),
b-(benzo-1H-1,2,3-triazol-1-yl)-3b-pyrrolidino-5a-
d
5.1.4.6. 16
androstan-17-one (8f). White solid. yield 51%. m.p. 173e175 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.88 (s, 3H, CH3-19), 1.12 (s, 3H, CH3-18),
2.83 (s, 3H, NeCH3), 4.57 (t, 1H, C16 H), 6.69 (t, 1H), 6.79 (d, 2H,
b
-(1H-1,2,4-triazol-1-yl)-3
b
-(N-methylphenylamino)-5
a-
-a
7.48e7.58(m, 2H), 8.07 (d, 1H, J ¼ 8.24Hz). Anal. Calcd. for
d
C29H40N4O: C 75.61, H 8.75, N 12.16; Found C 75.49, H 8.60, N 12.25.
-
a
J ¼ 8.28Hz), 7.24(t, 2H), 7.95 (s, 1H), 8.16 (s, 1H). Anal. Calcd. for
5.1.4.16. 16
b-(benzo-1H-1,2,3-triazol-1-yl)-3b-piperidino-5a-andro-
C28H38N4O: C 75.30, H 8.58, N 12.54; Found C 75.19, H 8.70, N 12.44.
stan-17-one (10b). White solid. yield 60%. m.p. 242e245 ꢀC; 1H

3668
H. Guo et al. / European Journal of Medicinal Chemistry 46 (2011) 3662e3674
NMR(400 MHz,CDCl3):
2.52 (s, 4H, NCH2), 4.98 (t, 1H, C16-a
d
0.84 (s, 3H, CH3-19), 1.23 (s, 3H, CH3-18),
H), 7.37 (t, 1H), 7.46e7.58(m,
5.1.4.25. 16
androstan-17-one (11d). White solid. yield 58%. m.p. 177e180 ꢀC;
1H NMR(400 MHz,CDCl3):
0.86 (s, 3H, CH3-19),1.11 (s, 3H, CH3-18),
2.26(s, 3H, NeCH3), 2.40e2.54 (br, 8H, NCH2), 4.73 (t, 1H, C16 H),
b-(1H-benzimidazol-1-yl)-3b-(4-Methylpiperazinyl)-5a-
2H), 8.07 (d, 1H, J ¼ 8.36Hz). Anal. Calcd. for C30H42N4O: C 75.91, H
d
8.92, N 11.80; Found C 75.80, H 8.77, N 11.92.
-a
7.29e7.31(m, 3H), 7.41e7.42 (m, 3H), 7.81 (s, 1H). Anal. Calcd. for
5.1.4.17. 16
rostan-17-one (10c). White solid. yield 60%. m.p. 218e220 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.86 (s, 3H, CH3-19), 1.24 (s, 3H, CH3-18),
2.57 (s, 4H, NCH2), 3.73 (s, 4H, OCH2), 4.98 (t, 1H, C16 H), 7.37 (t,
1H), 7.47e7.58(m, 2H), 8.07 (d, 1H, J ¼ 8.36Hz). Anal. Calcd. for
C29H40N4O2: C 73.07, H 8.46, N 11.75; Found C 73.19, H 8.60, N
11.63.
b
-(benzo-1H-1,2,3-triazol-1-yl)-3
b
-morpholino-5
a
-and
C31H44N4O: C 76.19, H 9.07, N 11.46; Found C 76.31, H 9.20, N 11.38.
d
5.1.4.26. 16
stan-17-one (11e). White solid. yield 58%. m.p. 203e206 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.87 (s, 3H, CH3-19), 1.10 (s, 3H, CH3-18),
2.52 (s, 4H, NCH2), 4.98 (t, 1H, C16 H), 7.29e7.31(m, 3H), 7.41e7.42
b-(1H-benzimidazol-1-yl)-3b-diethylamino-5a-andro-
-a
d
-a
(m, 3H), 7.82 (s, 1H). Anal. Calcd. for C30H43N3O: C 78.05, H 9.39, N
9.10; Found C 78.13, H 9.51, N 9.22.
5.1.4.18. 16
-androstan-17-one (10d). White solid. yield 60%. m.p.
215e218 ꢀC; 1H NMR(400 MHz,CDCl3):
0.85 (s, 3H, CH3-19),1.23 (s,
3H, CH3-18), 2.28(s, 3H, NeCH3), 2.47e2.63 (br, 8H, NCH2), 4.98 (t,
1H, C16
b-(benzo-1H-1,2,3-triazol-1-yl)-3b-(4-Methylpiperazinyl)-
5a
5.1.4.27. 16
androstan-17-one (11f). White solid. yield 58%. m.p. 212e214 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.89 (s, 3H, CH3-19), 1.03 (s, 3H, CH3-18), 2.79
(s, 3H, NeCH3), 5.02 (t, 1H, C16 H), 6.69 (t, 1H), 6.79 (d, 2H,
J ¼ 8.24Hz), 7.20e7.30 (m, 5H), 7.77 (d, 2H, J ¼ 6.96Hz), 8.06 (s, 1H).
Anal. Calcd. for C33H41N3O: C 79.96, H 8.34, N 8.48; Found C 79.88, H
8.21, N 8.54.
b-(1H-benzimidazol-1-yl)-3b-(N-methylphenylamino)-5a-
d
d
-a
H), 7.37 (t, 1H), 7.48e7.58(m, 2H), 8.06 (d, 1H, J ¼ 8.36Hz).
-a
Anal. Calcd. for C30H43N5O: C 73.58, H 8.85, N 14.30; Found C 73.44,
H 8.78, N 14.19.
5.1.4.19. 16
androstan-17-one (10e). White solid. yield 60%. m.p. 130e132 ꢀC;
1H NMR(400 MHz,CDCl3):
0.89 (s, 3H, CH3-19), 1.24 (s, 3H, CH3-
18), 2.52 (s, 4H, NCH2), 4.98 (t, 1H, C16 H), 7.37 (t, 1H),
7.48e7.58(m, 2H), 8.06 (d, 1H, J ¼ 8.36Hz). Anal. Calcd. for
C29H42N4O: C 75.28, H 9.15, N 12.11; Found C 75.18, H 9.30, N
12.01.
b-(benzo-1H-1,2,3-triazol-1-yl)-3b-diethylamino-5a-
5.1.4.28. 16
androstan-17-one (11g). White solid. yield 58%. m.p. 248e251 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.85 (s, 3H, CH3-19),1.01 (s, 3H, CH3-18), 2.21
(s, 3H, NeCH3), 3.58 (s, 2H, PhCH2), 4.73 (t,1H, C16 H), 7.29e7.30(m,
3H), 7.31 (s, 5H), 7.40e7.42 (m, 3H), 7.80 (s, 1H). Anal. Calcd. for
C34H43N3O: C 80.11, H 8.50, N 8.24; Found C 80.01, H 8.39, N 8.12.
b-(1H-benzimidazol-1-yl)-3b-(N-benzylmethylamino)-5a-
d
-a
d
-a
5.1.4.20. 16
-androstan-17-one (10f). White solid. yield 64%. m.p. 220e222 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.88 (s, 3H, CH3-19), 1.24 (s, 3H, CH3-18), 2.83
(s, 3H, NeCH3), 4.97 (t, 1H, C16-
7.24(t, 2H), 7.37 (t, 1H), 7.48e7.58(m, 2H), 8.06 (d, 1H, J ¼ 8.34Hz). Anal.
Calcd. for C32H40N4O: C 77.38, H 8.12, N 11.28; Found C 77.26, H 8.08, N
11.31.
b
-(benzo-1H-1,2,3-triazol-1-yl)-3
b
-(N-methylphenylamino)-
5.1.5. General procedure for the synthesis of compounds 12aeg,
13aeg, 14aeg, 15aeg
5a
d
To the suspension of compounds 8aeg, 9aeg, 10aeg, 11aeg
(1.00 mmol) in 20 ml of absolute methanol at 0 ꢀC, sodium boro-
hydride (1.20 mmol, 1.2 equiv) was added in small amounts over
a period of 4 h. The methanol was removed under reduced pres-
sure, 20 ml CH2Cl2 was added to the residue, the organic layer was
washed with saturated NaHCO3, brine and dried over anhydrous
Na2SO4, filtered. The filtrate was concentrated in vacuum to afford
a crude product. The product was purified by flash column chro-
matography to yield compound 12aeg, 13aeg, 14aeg, 15aeg.
aH), 6.69 (t, 1H), 6.79 (d, 2H, J ¼ 8.28Hz),
5.1.4.21. 16
-androstan-17-one (10g). White solid. yield 64%. m.p. 140e143 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.87 (s, 3H, CH3-19), 1.24 (s, 3H, CH3-18), 2.21
(s, 3H, NeCH3), 3.58 (s, 2H, PhCH2), 4.98 (t, 1H, C16- H), 7.24 (s, 1H),
b-(benzo-1H-1,2,3-triazol-1-yl)-3b-(N-benzylmethylamino)-
5a
d
a
7.31e7.37 (m, 4H), 7.46e7.58 (m, 2H), 7.86 (d, 2H, J ¼ 2.76Hz), 8.07 (d,
2H, J ¼ 8.32Hz). Anal. Calcd. for C33H42N4O: C 77.61, H 8.29, N 10.97;
Found C 77.80, H 8.41, N 10.88.
5.1.5.1. 16
ol (12a). White solid. yield 58%. m.p. 236e239 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.82 (s, 3H, CH3-19), 0.99 (s, 3H, CH3-18),
2.53 (s, 4H, NCH2), 3.83 (t, 1H, C17
b-(1H-1,2,4-triazol-1-yl)-3b-pyrrolidino-5a-androstan-17b-
d
-a
H, J ¼ 9.40Hz), 4.82 (q, 1H, C16
-
5.1.4.22. 16
17-one (11a). White solid. yield 64%. m.p. 133e136 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.84 (s, 3H, CH3-19), 1.09 (s, 3H, CH3-18),
2.57 (s, 4H, NCH2), 4.73 (t, 1H, C16 H), 7.29e7.32(m, 3H), 7.40e7.42
b
-(1H-benzimidazol-1-yl)-3
b
-pyrrolidino-5a-androstan-
aH), 7.96 (s, 1H), 8.10 (s, 1H). Anal. Calcd. for C25H40N4O: C 72.77, H
9.77, N 13.58; Found C 72.61, H 9.82, N 13.66.
d
-a
5.1.5.2. 16
b
-(1H-1,2,4-triazol-1-yl)-3
b
-piperidino-5
a
-androstan-17
b-
(m, 3H), 7.80 (s, 1H). Anal. Calcd. for C30H41N3O: C 78.39, H 8.99, N
9.14; Found C 78.41, H 8.82, N 9.30.
ol (12b). White solid. yield 58%. m.p. 162e165 ꢀC; 1H
NMR(400 MHz,CDCl3):
2.52 (s, 4H, NCH2), 3.85 (d, 1H, C17
d
0.80 (s, 3H, CH3-19), 0.99 (s, 3H, CH3-18),
H,), 4.82 (q, 1H, C16 H), 7.95 (s,
-a
-a
5.1.4.23. 16
17-one (11b). White solid. yield 60%. m.p. 219e222 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.85 (s, 3H, CH3-19), 1.08 (s, 3H, CH3-18),
2.53 (s, 4H, NCH2), 4.74 (t, 1H, C16 H), 7.29e7.31(m, 3H), 7.40e7.42
(m, 3H), 7.81 (s, 1H). Anal. Calcd. for C31H43N3O: C 78.60, H 9.15, N
8.87; Found C 78.50, H 9.24, N 8.98.
b
-(1H-benzimidazol-1-yl)-3
b
-piperidino-5
a
-androstan-
1H), 8.09 (s, 1H). Anal. Calcd. for C26H42N4O: C 73.20, H 9.92, N
13.13; Found C 73.11, H 9.86, N 13.28.
d
-a
5.1.5.3. 16
(12c). White solid. yield 58%. m.p. 162e165 ꢀC; 1H NMR
(400 MHz,CDCl3): 0.80 (s, 3H, CH3-19), 0.83 (s, 3H, CH3-18), 2.57 (s,
4H, NCH2), 3.71 (s, 4H, OCH2), 3.88 (d, 1H, C17-
H, J ¼ 5.04Hz), 5.06 (q,
1H, C16 H), 7.90 (s, 1H), 8.14 (s, 1H). Anal. Calcd. for C25H40N4O2: C
70.06 H 9.41, N 13.07; Found C 70.18, H 9.29, N 13.12.
b-(1H-1,2,4-triazol-1-yl)-3b-morpholino-5a-androstan-17b-ol
d
a
5.1.4.24. 16
17-one (11c). White solid. yield 58%. m.p. 209e212 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.84 (s, 3H, CH3-19), 1.09 (s, 3H, CH3-18),
2.57 (s, 4H, NCH2), 3.73 (s, 4H, OCH2), 4.74 (t, 1H, C16 H),
b
-(1H-benzimidazol-1-yl)-3
b
-morpholino-5a-androstan-
-a
d
-a
5.1.5.4. 16
androstan-17
1H NMR(400 MHz,CDCl3):
b
-(1H-1,2,4-triazol-1-yl)-3
-ol (12d). White solid. yield 49%. m.p. 259e262 ꢀC;
0.80 (s, 3H, CH3-19), 0.99 (s, 3H, CH3-
b-(4-Methylpiperazinyl)-5a-
7.29e7.31(m, 3H), 7.41e7.42 (m, 3H), 7.81 (s, 1H). Anal. Calcd. for
C30H41N3O2: C 75.75, H 8.69, N 8.83; Found C 75.62, H 8.88, N 8.78.
b
d

H. Guo et al. / European Journal of Medicinal Chemistry 46 (2011) 3662e3674
3669
18), 2.27 (s, 3H, NeCH3), 2.43e2.61 (br, 8H, NCH2), 3.84 (d, 2H, C17
-
5.1.5.13. 16
androstan-17
NMR(400 MHz,CDCl3):
2.78 (s, 3H, NeCH3), 3.85 (s,1H, C17
b
-(1H-1,2,3-triazol-1-yl)-3
-ol (13f). White solid. yield 61%. m.p. 198e202 ꢀC; 1H
0.88 (s, 3H, CH3-19), 1.01 (s, 3H, CH3-18),
H), 4.83 (q,1H, C16 H), 6.68 (t,
b-(N-methylphenylamino)-5a-
a
H, J ¼ 9.08Hz), 4.82 (q, 1H, C16
-a
H), 7.96 (s, 1H), 8.11 (s, 1H). Anal.
b
Calcd. for C26H43N5O: C 70.71, H 9.81, N 15.86; Found C 70.66, H
9.73, N 15.71.
d
-a
-a
1H), 6.78 (d, 2H, J ¼ 8.20Hz), 7.21 (t, 2H), 7.97 (s, 1H), 8.10 (s, 1H).
Anal. Calcd. for C28H40N4O: C 74.96, H 8.99, N 12.49; Found C 74.88,
H 8.83, N 12.60.
5.1.5.5. 16
17
-ol (12e). White solid. yield 60%. m.p. 215e219 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.84 (s, 3H, CH3-19), 0.99 (s, 3H, CH3-18),
2.62 (q, 4H, NCH2), 3.83 (s, 1H, C17 H), 4.82 (q, 1H, C16 H), 7.95 (s,
1H), 8.09 (s, 1H). Anal. Calcd. for C25H42N4O: C 72.42, H 10.21, N
13.51; Found C 72.33, H 10.04, N 13.62.
b-(1H-1,2,4-triazol-1-yl)-3b-diethylamino-5a-androstan-
b
d
-
a
-
a
5.1.5.14. 16
androstan-17
NMR(400 MHz,CDCl3):
2.20 (s, 3H, NeCH3), 3.57 (s, 2H, PhCH2), 3.86 (s, 1H, C17
1H, C16 H), 7.31 (t, 5H), 7.97 (s, 1H), 8.10 (s, 1H). Anal. Calcd. for
C29H42N4O: C 75.28, H 9.15, N 12.11; Found C 75.09, H 9.23, N 12.01.
b
-(1H-1,2,3-triazol-1-yl)-3
-ol (13g). White solid. yield 53%. m.p. 220e223 ꢀC; 1H
0.83 (s, 3H, CH3-19), 1.00 (s, 3H, CH3-18),
H), 4.84 (t,
b-(N-benzylmethylamino)-5a-
b
d
-a
5.1.5.6. 16
androstan-17
NMR(400 MHz,CDCl3):
2.78 (s, 3H, NeCH3), 3.85 (s,1H, C17
b
-(1H-1,2,4-triazol-1-yl)-3
-ol (12f). White solid. yield 61%. m.p. 198e202 ꢀC; 1H
0.88 (s, 3H, CH3-19), 1.01 (s, 3H, CH3-18),
H), 4.83 (q,1H, C16 H), 6.68 (t,
b
-(N-methylphenylamino)-5
a
-
-a
b
d
-
a
-
a
5.1.5.15. 16
stan-17
-ol (14a). White solid. yield 60%. m.p. 259e262 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.84 (s, 3H, CH3-19), 1.11 (s, 3H, CH3-18),
2.56 (s, 4H, NCH2), 4.03 (t, 1H, C17 H), 5.20 (q, 1H, C16 H), 7.37 (t,
b-(benzo-1H-1,2,3-triazol-1-yl)-3b-pyrrolidino-5a-andro-
1H), 6.78 (d, 2H, J ¼ 8.20Hz), 7.21 (t, 2H), 7.97 (s, 1H), 8.10 (s, 1H).
Anal. Calcd. for C28H40N4O: C 74.96, H 8.99, N 12.49; Found C 74.88,
H 8.83, N 12.60.
b
d
-a
-a
1H,), 7.47e7.54 (m, 2H), 8.05 (d, 1H, J ¼ 8.40Hz). Anal. Calcd. for
5.1.5.7. 16
androstan-17
1H NMR(400 MHz,CDCl3):
18), 2.20 (s, 3H, NeCH3), 3.57 (s, 2H, PhCH2), 3.86 (s, 1H, C17
4.84 (t, 1H, C16 H), 7.31 (t, 5H), 7.97 (s, 1H), 8.10 (s, 1H). Anal. Calcd.
b
-(1H-1,2,4-triazol-1-yl)-3
-ol (12g). White solid. yield 53%. m.p. 206e208 ꢀC;
0.83 (s, 3H, CH3-19), 1.00 (s, 3H, CH3-
H),
b
-(N-benzylmethylamino)-5
a
-
C29H42N4O: C 75.28, H 9.15, N 12.11; Found C 75.17, H 9.21, N 11.98.
b
d
5.1.5.16. 16
stan-17
-ol (14b). White solid. yield 61%. m.p. 263e266 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.86 (s, 3H, CH3-19), 1.08 (s, 3H, CH3-18),
2.53 (s, 4H, NCH2), 4.02 (t, 1H, C17 H), 5.18 (q, 1H, C16 H), 7.37 (t,
b-(benzo-1H-1,2,3-triazol-1-yl)-3b-piperidino-5a-andro-
-a
b
-a
d
for C29H42N4O: C 75.28, H 9.15, N 12.11; Found C 75.09, H 9.23, N
12.01.
-a
-a
1H,), 7.47e7.54 (m, 2H), 8.03 (d, 1H, J ¼ 8.36Hz). Anal. Calcd. for
C30H44N4O: C 75.59, H 9.30, N 11.75; Found C 75.67, H 9.14, N 11.69.
5.1.5.8. 16
(13a). White solid. yield 58%. m.p. 236e239 ꢀC; 1H NMR
(400 MHz,CDCl3): 0.82 (s, 3H, CH3-19), 0.99 (s, 3H, CH3-18), 2.53 (s, 4H,
NCH2), 3.83 (t, 1H, C17- H), 7.96 (s, 1H),
8.10 (s,1H). Anal. Calcd. for C25H40N4O: C 72.77, H 9.77, N 13.58; Found C
72.61, H 9.82, N 13.66.
b-(1H-1,2,3-triazol-1-yl)-3b-pyrrolidino-5a-androstan-17b-ol
5.1.5.17. 16
stan-17
-ol (14c). White solid. yield 57%. m.p. 257e259 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.83 (s, 3H, CH3-19), 1.12 (s, 3H, CH3-18),
2.57 (s, 4H, NCH2), 3.72 (s, 4H, OCH2), 4.03 (t,1H, C17 H), 5.20 (q,1H,
C16
b-(benzo-1H-1,2,3-triazol-1-yl)-3b-morpholino-5a-andro-
d
b
a
H, J ¼ 9.40Hz), 4.82 (q, 1H, C16-
a
d
-a
-a
H), 7.36 (t, 1H,), 7.47e7.54 (m, 2H), 8.05 (d, 1H, J ¼ 8.36Hz).
Anal. Calcd. for C29H42N4O2: C 72.77, H 8.84, N 11.70; Found C 72.84,
H 8.85, N 11.64.
5.1.5.9. 16
17
-ol (13b). White solid. yield 58%. m.p. 162e165 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.80 (s, 3H, CH3-19), 0.99 (s, 3H, CH3-18),
2.52 (s, 4H, NCH2), 3.85 (d, 1H, C17 H,), 4.82 (q, 1H, C16 H), 7.95 (s,
b-(1H-1,2,3-triazol-1-yl)-3b-piperidino-5a-androstan-
b
d
5.1.5.18. 16
androstan-17
295 ꢀC; 1H NMR(400 MHz,CDCl3):
CH3-18), 2.27 (s, 3H, NeCH3), 2.42e2.61 (br, 8H, NCH2), 4.03 (s, 1H, C17-
H), 5.20 (q, 1H, C16- H), 7.35 (t, 1H,), 7.46e7.54 (m, 2H), 8.02 (d, 1H,
b
-(benzo-1H-1,2,3-triazol-1-yl)-3
-ol (14d). White solid. yield 65%. m.p. 292e
0.83 (s, 3H, CH3-19), 1.12 (s, 3H,
b-(4-Methylpiperazinyl)-5a-
-a
-a
b
1H), 8.09 (s, 1H). Anal. Calcd. for C26H42N4O: C 73.20, H 9.92, N
13.13; Found C 73.11, H 9.86, N 13.18.
d
a
a
5.1.5.10. 16
17
-ol (13c). White solid. yield 58%. m.p. 162e165 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.80 (s, 3H, CH3-19), 0.83 (s, 3H, CH3-18),
2.57 (s, 4H, NCH2), 3.71 (s, 4H, OCH2), 3.88 (d, 1H, C17 H,
H), 7.90 (s, 1H), 8.14 (s, 1H). Anal.
b
-(1H-1,2,3-triazol-1-yl)-3
b
-morpholino-5
a
-androstan-
J ¼ 8.36Hz). Anal. Calcd. for C30H45N5O: C 73.28, H 9.22, N 14.24; Found
b
C 73.19, H 9.09, N 14.21.
d
-a
5.1.5.19. 16
stan-17
-ol (14e). White solid. yield 70%. m.p. 240e243 ꢀC; H NMR
(400 MHz,CDCl3): 0.83 (s, 3H, CH3-19), 1.12 (s, 3H, CH3-18), 2.52 (s,
4H, NCH2), 4.02 (t, 1H, C17 H), 5.20 (q, 1H, C16 H), 7.36 (t, 1H,),
b-(benzo-1H-1,2,3-triazol-1-yl)-3b-diethylamino-5a-andro-
1
J ¼ 5.04Hz), 5.06 (q, 1H, C16
-a
b
Calcd. for C25H40N4O2: C 70.06 H 9.41, N 13.07; Found C 70.18, H
9.29, N 13.12.
d
-a
-a
7.47e7.54 (m, 2H), 8.04 (d, 1H, J ¼ 8.40Hz). Anal. Calcd. for C29H44N4O:
5.1.5.11. 16
androstan-17
1H NMR(400 MHz,CDCl3):
18), 2.27 (s, 3H, NeCH3), 2.43e2.61 (br, 8H, NCH2), 3.84 (d, 2H, C17
H), 7.96 (s, 1H), 8.11 (s, 1H). Anal.
b
-(1H-1,2,3-triazol-1-yl)-3
-ol (13d). White solid. yield 49%. m.p. 259e262 ꢀC;
0.80 (s, 3H, CH3-19), 0.99 (s, 3H, CH3-
b
-(4-Methylpiperazinyl)-5
a
-
C 74.96, H 9.54, N 12.06; Found C 74.88, H 9.62, N 12.03.
b
d
5.1.5.20. 16
ino)-5 -androstan-17
243e246 ꢀC; 1H NMR(400 MHz,CDCl3):
3H, CH3-18), 2.85 (s, 3H, NeCH3), 4.04 (t,1H, C17
H), 6.69 (t, 1H), 6.68e6.77 (m, 1H), 6.85e6.96 (m, 2H), 7.24(t, 2H),
b
-(benzo-1H-1,2,3-triazol-1-yl)-3
-ol (14f). White solid. yield 70%. m.p.
0.88 (s, 3H, CH3-19), 1.12 (s,
H), 5.21 (q,1H, C16
b-(N-methylphenylam-
-
a
b
a
H, J ¼ 9.08Hz), 4.82 (q, 1H, C16
-a
d
Calcd. for C26H43N5O: C 70.71, H 9.81, N 15.86; Found C 70.66, H
9.73, N 15.71.
-a
-
a
7.36 (t,1H), 7.47e7.55(m, 2H), 8.05 (d,1H, J ¼ 8.32Hz). Anal. Calcd. for
5.1.5.12. 16
17
-ol (13e). White solid. yield 60%. m.p. 215e219 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.84 (s, 3H, CH3-19), 0.99 (s, 3H,
CH3-18), 2.62 (q, 4H, NCH2), 3.83 (s, 1H, C17 H), 4.82 (q, 1H,
C16 H), 7.95 (s, 1H), 8.09 (s, 1H). Anal. Calcd. for
b
-(1H-1,2,3-triazol-1-yl)-3
b
-diethylamino-5
a
-androstan-
C32H42N4O: C 77.07, H 8.49, N 11.23; Found C 77.14, H 8.37, N 11.32.
b
d
5.1.5.21. 16
-androstan-17
NMR(400 MHz,CDCl3):
(s, 3H, NeCH3), 3.59 (s, 2H, PhCH2), 4.05 (s, 1H, C17-
H), 7.31 (t, 5H), 7.37 (t, 1H,), 7.47e7.54 (m, 2H), 8.06 (d,1H, J ¼ 8.40Hz).
b
-(benzo-1H-1,2,3-triazol-1-yl)-3
-ol (14g). White solid. yield 52%. m.p. 231e234 ꢀC; 1H
0.85 (s, 3H, CH3-19), 1.12 (s, 3H, CH3-18), 2.22
H), 5.21 (t, 1H, C16-
b-(N-benzylmethylamino)-
-
a
5a
b
-
a
d
C25H42N4O: C 72.42, H 10.21, N 13.51; Found C 72.33, H
10.04, N 13.62.
a
a

3670
H. Guo et al. / European Journal of Medicinal Chemistry 46 (2011) 3662e3674
Anal. Calcd. for C33H44N4O: C 77.30, H 8.65, N 10.93; Found C 77.41, H
8.58, N 10.82.
(monitored by TLC). The reaction was concentrated in reduced
pressure. The crude product was dissolved in CH2Cl2, the organic
layer was washed with brine and dried over anhydrous Na2SO4,
filtered. The filtrate was concentrated in reduced pressure to
afford a crude product. The products were purified by flash
column chromatography to yield compound 16aeg, 17aeg, 18aeg,
19aeg.
5.1.5.22. 16
17
-ol (15a). White solid. yield 59%. m.p. 279e282 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.83 (s, 3H, CH3-19), 0.93 (s, 3H, CH3-18),
2.55 (s, 4H, NCH2), 4.01 (d, 1H, C17
H, J ¼ 9.16Hz), 4.79 (q, 1H, C16
b-(1H-benzimidazol-1-yl)-3b-pyrrolidino-5a-androstan-
b
d
-a
-
a
H), 7.24 (t, 2H), 7.33 (d,1H, J ¼ 7.28Hz), 7.67 (d,1H, J ¼ 7.32Hz), 8.04
(s, 1H). Anal. Calcd. for C30H43N3O: C 78.05, H 9.39, N 9.10; Found C
78.12, H 9.56, N 9.24.
5.1.6.1. 16
acetate (16a). White solid. Yield 96%. m.p. 167e170 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.83 (s,3H, CH3-19), 1.11 (s,3H, CH3-18), 1.72 (s,
3H, OCOCH3), 2.56 (s,4H, NCH2), 4.70 (d, 1H, C17-
H, J ¼ 9.12Hz), 5.01 (q,
1H, C16- 12.28,
H), 7.90 (s, 1H), 7.92 (s, 1H). 13C NMR(150 MHz,CDCl3):
b-(1H-1,2,4-triazol-1-yl)-3b-pyrrolidino-5a-androstan-17b-yl
d
5.1.5.23. 16
17
-ol (15b). White solid. yield 56%. m.p. 273e275 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.81 (s, 3H, CH3-19), 0.93 (s, 3H, CH3-18),
2.52 (s, 4H, NCH2), 4.01 (d, 1H, C17
H, J ¼ 9.20Hz), 4.82 (q, 1H, C16
b
-(1H-benzimidazol-1-yl)-3
b
-piperidino-5a-androstan-
a
b
a
d
d
12.57, 20.08, 20.34, 23.13, 27.74, 28.60, 31.30, 31.86, 34.24, 34.46, 35.84,
37.34, 37.78, 42.13, 45.15, 48.41, 51.69, 54.31, 58.12, 64.17, 81.57, 145.02,
151.13, 170.88. Anal. Calcd. for C27H42N4O2: C 71.33, 9.31, N 12.32; Found
C 71.18, H 9.55, N 12.24. MS (m/z):455[M þ 1]þ.
-a
-
a
H), 7.22 (t, 2H), 7.35 (d,1H, J ¼ 7.32Hz), 7.70 (d,1H, J ¼ 7.30Hz), 8.05
(s, 1H). Anal. Calcd. for C31H45N3O: C 78.27, H 9.53, N 8.83; Found C
78.41, H 9.36, N 8.71.
5.1.6.2. 16
acetate (16b). White solid. Yield 93%. m.p. 197e200 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.81 (s,3H, CH3-19), 1.11 (s,3H, CH3-18), 1.72 (s,
3H, OCOCH3), 2.51 (s,4H, NCH2), 4.69 (d, 1H, C17-
H, J ¼ 9.08Hz), 5.01 (q,
1H, C16- 12.38,
H), 7.90 (s, 1H), 7.92 (s, 1H). 13C NMR(150 MHz,CDCl3):
b-(1H-1,2,4-triazol-1-yl)-3b-piperidino-5a-androstan-17b-yl
5.1.5.24. 16
17
-ol (15c). White solid. yield 56%. m.p. 278e280 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.82 (s, 3H, CH3-19), 0.94 (s, 3H, CH3-18),
2.55 (s, 4H, NCH2), 3.71 (s, 4H, OCH2), 4.02 (d, 1H, C17 H,
b-(1H-benzimidazol-1-yl)-3b-morpholino-5a-androstan-
b
d
d
a
-a
a
d
J ¼ 9.20Hz), 4.79 (q, 1H, C16
-
a
H), 7.21 (t, 2H), 7.35 (d, 1H, J ¼ 7.20Hz),
12.60, 20.11, 20.43, 23.95, 24.73, 26.21, 28.69, 30.49, 31.88, 34.28, 36.05,
37.80, 37.94, 42.17, 45.97, 48.43, 50.28, 54.34, 58.16, 64.50, 81.61, 145.05,
151.16, 170.92. Anal. Calcd. for C28H44N4O2: C 71.76, H 9.46, N 11.95;
Found C 71.69, H 9.60, N 11.87. MS (m/z):469[M þ 1]þ.
7.66 (d, 1H, J ¼ 7.48Hz), 8.03 (s, 1H). Anal. Calcd. for C30H43N3O2: C
75.43, H 9.07, N 8.80; Found C 75.32, H 9.16, N 8.68.
5.1.5.25. 16
androstan-17
1H NMR(400 MHz,CDCl3):
18), 2.24 (s, 3H, NeCH3), 2.43e2.59 (br, 8H, NCH2), 4.02 (d, 1H, C17
H), 7.21 (t, 2H), 7.33 (d, 1H,
b
-(1H-benzimidazol-1-yl)-3
-ol (15d). White solid. yield 56%. m.p. 279e281 ꢀC;
0.81 (s, 3H, CH3-19), 0.94 (s, 3H, CH3-
b-(4-Methylpiperazinyl)-5a-
b
5.1.6.3. 16
17
-yl acetate (16c). White solid. Yield 92%. m.p. 183e186 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.82 (s,3H, CH3-19), 1.12 (s,3H, CH3-18),
1.72 (s, 3H, OCOCH3), 2.56 (s,4H, NCH2), 3.72 (s, 4H, OCH2), 4.69 (d,
1H, C17 H), 7.90 (s, 1H), 7.92 (s, 1H).
H, J ¼ 9.12Hz), 5.01 (q, 1H, C16
13C NMR(150 MHz,CDCl3):
12.33, 12.59, 20.10, 20.40, 24.37, 28.67,
b-(1H-1,2,4-triazol-1-yl)-3b-morpholino-5a-androstan-
d
b
-
d
a
H, J ¼ 9.16Hz), 4.79 (q, 1H, C16
-a
J ¼ 7.36Hz), 7.66 (d, 1H, J ¼ 7.32Hz), 8.06 (s, 1H). Anal. Calcd. for
-a
-a
C31H46N4O: C 75.87, H 9.45, N 11.42; Found C 75.72, H 9.31, N 11.60.
d
31.00, 31.30, 31.84, 34.24, 35.96, 37.64, 37.76, 42.14, 45.61, 48.39,
49.97, 54.28, 58.12, 63.98, 67.26, 81.56, 145.04, 151.13, 170.90. Anal.
Calcd. for C27H42N4O3: C 68.90, H 8.99, N 11.90; Found C 68.78, H
9.16, N 11.83. MS (m/z):471[M þ 1]þ.
5.1.5.26. 16
17
-ol (15e). White solid. yield 56%. m.p. 227e230 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.82 (s, 3H, CH3-19), 0.93 (s, 3H, CH3-18), 2.56
(q, 4H, NCH2), 4.02 (d, 1H, C17- H), 7.20
(t, 2H), 7.40 (d, 1H, J ¼ 7.96Hz), 7.64 (d, 1H, J ¼ 7.76Hz), 7.97 (s, 1H). Anal.
Calcd. for C30H45N3O: C 77.71, H 9.78, N 9.06; Found C 77.64, H 9.89, N
9.17.
b-(1H-benzimidazol-1-yl)-3b-diethylamino-5a-androstan-
b
d
a
H, J ¼ 9.22Hz), 4.78 (q, 1H, C16
-a
5.1.6.4. 16
androstan-17
208e210 ꢀC; 1H NMR(400 MHz,CDCl3):
(s,3H, CH3-18), 1.72 (s, 3H, OCOCH3), 2.28 (s, 3H, NeCH3),
2.46e2.61 (br, 8H, NCH2), 4.69 (d, 1H, C17
H, J ¼ 9.12Hz), 5.01 (q,
1H, C16
H), 7.90 (s, 1H), 7.92 (s, 1H). 13C NMR(150 MHz,CDCl3):
12.38, 12.60, 20.13, 20.42, 24.45, 28.69, 31.22, 31.33, 31.87, 34.27,
b
-(1H-1,2,4-triazol-1-yl)-3
-yl acetate (16d). White solid. Yield 92%. m.p.
0.81 (s,3H, CH3-19), 1.12
b-(4-Methylpiperazinyl)-5a-
b
d
5.1.5.27. 16
androstan-17
NMR(400 MHz,CDCl3):
3H, NeCH3), 4.01 (d,1H, C17-
1H), 6.96 (d, 2H, J ¼ 8.20Hz), 7.21 (t, 2H), 7.24 (t, 2H), 7.35 (d, 1H,
J ¼ 7.62Hz), 7.70 (d, 1H, J ¼ 7.44Hz), 8.07 (s, 1H). Anal. Calcd. for
C33H43N3O: C 79.63, H 8.71, N 8.44; Found C 79.80, H 8.65, N 8.52.
b
-(1H-benzimidazol-1-yl)-3
-ol (15f). White solid. yield 59%. m.p. 264e266 ꢀC; 1H
0.88 (s, 3H, CH3-19), 0.94 (s, 3H, CH3-18), 2.79 (s,
H), 6.79 (t,
b
-(N-methylphenylamino)-5
a
-
-a
b
-a
d
d
a
H, J ¼ 9.22Hz), 4.82 (q,1H, C16
-a
35.98, 37.79, 42.16, 45.76, 46.00, 48.42, 49.25, 54.32, 55.46, 58.15,
63.74, 81.61, 145.06, 151.19, 170.95. Anal. Calcd. for C28H45N5O2: C
69.53, H 9.38, N 14.48; Found C 69.62, H 9.33, N 14.63. MS
(m/z):484[M þ 1]þ.
5.1.5.28. 16
androstan-17
NMR(400 MHz,CDCl3):
(s, 3H, NeCH3), 3.57 (s, 2H, PhCH2), 4.01 (d, 1H, C17-
(q, 1H, C16-
b
-(1H-benzimidazol-1-yl)-3
-ol (15g). White solid. yield 56%. m.p. 214e216 ꢀC; 1H
0.84 (s, 3H, CH3-19), 0.94 (s, 3H, CH3-18), 2.20
H, J ¼ 9.16Hz), 4.82
b
-(N-benzylmethylamino)-5
a
-
5.1.6.5. 16
17
-yl acetate (16e). White solid. Yield 92%. m.p. 115e118 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.82 (s,3H, CH3-19), 1.12 (s,3H, CH3-18),
1.72 (s, 3H, OCOCH3), 2.64 (q, 4H, NCH2), 4.70 (d, 1H, C17 H,
J ¼ 9.12Hz), 5.01 (q, 1H, C16
H), 7.90 (s, 1H), 7.93 (s, 1H). 13C
NMR(150 MHz,CDCl3): 12.38, 12.60, 13.20, 20.10, 20.43, 24.20,
28.69, 30.80, 31.33, 31.85, 34.27, 36.04, 37.79, 37.88, 42.16, 43.57,
45.97, 48.42, 54.31, 58.16, 59.64, 81.59, 145.04, 151.14, 170.93. Anal.
Calcd. for C27H44N4O2: C 71.01, H 9.71, N 12.27; Found C 71.20, H
9.66, N 12.09. MS (m/z):457[M þ 1]þ.
b-(1H-1,2,4-triazol-1-yl)-3b-diethylamino-5a-androstan-
b
b
d
d
a
-a
aH), 7.21 (t, 2H), 7.31 (m, 6H), 7.66 (d, 1H, J ¼ 7.28Hz), 8.07 (s,
-a
1H). Anal. Calcd. for C34H45N3O: C 79.80, H 8.86, N 8.21; Found C 79.70,
H 8.93, N 8.08.
d
5.1.6. General procedure for the synthesis of compounds 16aeg,
17aeg, 18aeg, 19aeg
To a solution of compounds 12aeg, 13aeg, 14aeg, 15aeg
(2.00 mmol) in CH2Cl2 was added acetic anhydride (4.00 mmol,
2.0 equiv) and 4-dimethylaminopyridine (2.00 mmol, 1.0 equiv).
The mixture was stirred at 40 ꢀC until the reaction was completed
5.1.6.6. 16
androstan-17
179e182 ꢀC; 1H NMR(400 MHz,CDCl3):
b
-(1H-1,2,4-triazol-1-yl)-3
-yl acetate (16f). White solid. Yield 92%. m.p.
0.88 (s,3H, CH3-19), 1.13
b-(N-methylphenylamino)-5a-
b
d

H. Guo et al. / European Journal of Medicinal Chemistry 46 (2011) 3662e3674
3671
(s,3H, CH3-18), 1.73 (s, 3H, OCOCH3), 2.78 (s, 3H, NeCH3), 4.71 (d,
1H, C17 H), 6.68 (t,1H), 6.79 (d, 2H,
H, J ¼ 9.12Hz), 5.00 (q,1H, C16
8.20Hz), 7.22 (t, 2H), 7.91 (s, 1H), 7.95 (s, 1H). 13C
NMR(150 MHz,CDCl3): 12.45, 12.61, 20.11, 20.46, 24.91, 28.50,
31.31, 31.40, 31.52, 31.79, 34.27, 35.84, 37.76, 37.89, 42.17, 46.12,
48.37, 54.26, 58.21, 58.32, 81.55, 113.32, 116.42, 129.07, 145.08,
150.14, 150.91, 170.95. Anal. Calcd. for C30H42N4O2: C 73.43, H 8.63,
N 11.42; Found C 73.31, H 8.59, N 11.52. MS (m/z): 491[M þ 1]þ.
5.1.6.12. 16
17
NMR(400 MHz,CDCl3):
1.72 (s, 3H, OCOCH3), 2.63 (q, 4H, NCH2), 4.71 (d, 1H, C17
J ¼ 9.14Hz), 5.01 (q, 1H, C16
H), 7.90 (s, 1H), 7.93 (s, 1H). 13C
NMR(150 MHz,CDCl3): 12.37, 12.60, 13.21, 20.08, 20.43, 24.21,
28.70, 30.81, 31.32, 31.86, 34.27, 36.04, 37.80, 37.89, 42.15, 43.57,
45.97, 48.42, 54.30, 58.16, 59.64, 81.61, 145.01, 151.15, 170.93. Anal.
Calcd. for C27H44N4O2: C 71.01, H 9.71, N 12.27; Found C 71.09, H
9.80, N 12.40. MS (m/z):457[M þ 1]þ.
b
-(1H-1,2,3-triazol-1-yl)-3
b
-diethylamino-5
a
-androstan-
-a
-a
b
-yl acetate (17e). White solid. Yield 90%. m.p. 140e143 ꢀC; 1H
J
¼
d
0.82 (s,3H, CH3-19), 1.11 (s,3H, CH3-18),
H,
d
-a
-a
d
5.1.6.7. 16
androstan-17
98e100 ꢀC; 1H NMR(400 MHz,CDCl3):
(s,3H, CH3-18),1.72 (s, 3H, OCOCH3), 2.21 (s, 3H, NeCH3), 3.59 (s, 2H,
b
-(1H-1,2,4-triazol-1-yl)-3
-yl acetate (16g). White solid. Yield 88%. m.p.
0.83 (s,3H, CH3-19), 1.12
b-(N-benzylmethylamino)-5a-
b
5.1.6.13. 16
androstan-17
187e190 ꢀC; 1H NMR(400 MHz,CDCl3):
(s,3H, CH3-18), 1.72 (s, 3H, OCOCH3), 2.78 (s, 3H, NeCH3), 4.71 (d,
1H, C17 H), 6.68 (t, 1H), 6.79 (d, 2H,
H, J ¼ 9.16Hz), 5.01 (q, 1H, C16
8.20Hz), 7.22 (t, 2H), 7.91 (s, 1H), 7.95 (s, 1H). 13C
NMR(150 MHz,CDCl3): 12.40, 12.60, 20.09, 20.46, 24.92, 28.50,
b
-(1H-1,2,3-triazol-1-yl)-3
-yl acetate (17f). White solid. Yield 92%. m.p.
0.84 (s,3H, CH3-19), 1.13
b-(N-methylphenylamino)-5a-
d
b
d
PhCH2), 4.69 (d, 1H, C17
-
a
H, J ¼ 9.08Hz), 5.01 (q, 1H, C16
-aH),
7.16e7.37(m, 6H), 7.92 (d, 2H, J ¼ 8.60Hz).13C NMR(150 MHz,CDCl3):
-a
-a
d
12.41, 12.63, 20.16, 20.47, 23.66, 28.70, 30.20, 31.35, 31.85, 34.28,
J
¼
36.07, 37.48, 37.76, 37.80, 42.17, 45.83, 48.42, 54.26, 57.75, 58.16,
62.34, 81.62, 126.29, 127.02, 128.28, 129.09, 138.98, 145.10, 151.19,
170.97. Anal. Calcd. for C31H44N4O2: C 73.77, H 8.79, N 11.10; Found C
73.83, H 8.62, N 11.25. MS (m/z):505[M þ 1]þ.
d
31.30, 31.42, 31.52, 31.80, 34.27, 35.85, 37.74, 37.89, 42.20, 46.12,
48.38, 54.27, 58.20, 58.32, 81.53, 113.33, 116.43, 129.09, 145.08,
150.14, 150.91, 170.94. Anal. Calcd. for C30H42N4O2: C 73.43, H 8.63,
N 11.42; Found C 73.29, H 8.55, N 11.57. MS (m/z):491[M þ 1]þ.
5.1.6.8. 16
acetate (17a). White solid. Yield 85%. m.p. 177e180 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.84 (s,3H, CH3-19), 1.11 (s,3H, CH3-18), 1.72 (s,
3H, OCOCH3), 2.60 (s, 4H, NCH2), 4.70 (d,1H, C17-
H, J ¼ 9.12Hz), 5.01 (q,
1H, C16- 12.30,
H), 7.90 (s, 1H), 7.92 (s, 1H). 13C NMR(150 MHz,CDCl3):
b-(1H-1,2,3-triazol-1-yl)-3b-pyrrolidino-5a-androstan-17b-yl
5.1.6.14. 16
androstan-17
139e142 ꢀC; 1H NMR(400 MHz,CDCl3):
(s,3H, CH3-18),1.71 (s, 3H, OCOCH3), 2.22 (s, 3H, NeCH3), 3.60 (s, 2H,
b
-(1H-1,2,3-triazol-1-yl)-3
-yl acetate (17g). White solid. Yield 92%. m.p.
0.84 (s,3H, CH3-19), 1.12
b-(N-benzylmethylamino)-5a-
d
b
a
d
a
d
12.62, 20.14, 20.39, 23.19, 27.14, 28.59, 31.34, 31.85, 33.81, 34.26, 35.86,
37.28, 37.80, 42.18, 45.19, 48.45, 51.56, 54.27, 58.18, 64.24, 81.60, 145.04,
151.19, 171.00. Anal. Calcd. for C27H42N4O2: C 71.33, H 9.31, N 12.32;
Found C 71.19, H 9.42, N 12.18. MS (m/z):455[M þ 1]þ.
PhCH2), 4.70 (d, 1H, C17
-
a
H, J ¼ 9.06Hz), 5.00 (q, 1H, C16
-aH),
7.13e7.35(m, 6H), 7.91 (d, 2H, J ¼ 8.64Hz).13C NMR(150 MHz,CDCl3):
d
12.41, 12.62, 20.15, 20.48, 23.66, 28.70, 30.20, 31.35, 31.85, 34.28,
36.07, 37.48, 37.76, 37.78, 42.17, 45.83, 48.41, 54.26, 57.75, 58.14,
62.33, 81.62, 126.26, 127.04, 128.28, 129.10, 138.97, 145.10, 151.15,
170.95. Anal. Calcd. for C31H44N4O2: C 73.77, H 8.79, N 11.10; Found C
73.60, H 8.86, N 11.07. MS (m/z):505[M þ 1]þ.
5.1.6.9. 16
acetate (17b). White solid. Yield 88%. m.p. 189e192 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.81 (s,3H, CH3-19), 1.11 (s,3H, CH3-18), 1.72
(s, 3H, OCOCH3), 2.53 (s, 4H, NCH2), 4.69 (d, 1H, C17-
H, J ¼ 9.08Hz),
5.01 (q, 1H, C16-
H), 7.90 (s, 1H), 7.92 (s, 1H). 13C NMR(150 MHz,CDCl3):
12.37, 12.60, 20.11, 20.43, 23.82, 24.63, 26.04, 28.67, 30.36, 31.33,
b-(1H-1,2,3-triazol-1-yl)-3b-piperidino-5a-androstan-17b-yl
d
a
5.1.6.15. 16
tan-17
-yl acetate (18a). White solid. Yield 87%. m.p. 243e246 ꢀC;
1H NMR(400 MHz,CDCl3):
0.86 (s,3H, CH3-19), 1.18 (s,3H, CH3-18),
1.23 (s, 3H, OCOCH3), 2.58 (s, 4H, NCH2), 5.02 (d, 1H, C17 H,
b-(benzo-1H-1,2,3-triazol-1-yl)-3b-pyrrolidino-5a-andros
a
b
d
d
31.86, 34.27, 36.03, 37.79, 37.89, 42.17, 45.92, 48.42, 50.21, 54.31, 58.16,
64.49, 81.60, 145.05, 151.16, 170.93. Anal. Calcd. for C28H44N4O2: C
71.76, H 9.46, N 11.95; Found C 71.67, H 9.54, N 11.88. MS (m/z):469
[M þ 1]þ.
-a
J ¼ 9.08Hz), 5.28 (q, 1H, C16
-aH), 7.33 (t, 1H), 7.40e7.47 (m, 2H), 8.03
(d, 1H, J ¼ 8.28Hz). 13C NMR(150 MHz,CDCl3):
d 12.34, 12.70, 19.79,
20.36, 23.16, 27.76, 28.67, 31.35, 31.98, 34.41, 34.49, 35.91, 37.41,
37.84, 42.77, 45.23, 48.76, 51.67, 54.42, 56.74, 64.18, 80.51, 109.97,
119.56, 123.70, 126.95, 134.07, 145.48, 170.56. Anal. Calcd. for
C31H44N4O2: C 73.77, H 8.79, N 11.10; Found C 73.68, H 8.81, N 11.21.
MS (m/z):505[M þ 1]þ.
5.1.6.10. 16
17
-yl acetate (17c). White solid. Yield 93%. m.p. 168e170 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.82 (s,3H, CH3-19), 1.12 (s,3H, CH3-18),
1.72 (s, 3H, OCOCH3), 2.56 (s, 4H, NCH2), 3.72 (s, 4H, OCH2), 4.70
(d, 1H, C17 H), 7.90 (s, 1H), 7.92
H, J ¼ 9.16Hz), 5.01 (q, 1H, C16
(s, 1H). 13C NMR(150 MHz,CDCl3):
12.36, 12.62, 20.14, 20.44,
b-(1H-1,2,3-triazol-1-yl)-3b-morpholino-5a-androstan-
b
d
-
a
-a
5.1.6.16. 16
stan-17
-yl acetate (18b). White solid. Yield 93%. m.p. 271e273 ꢀC;
1H NMR(400 MHz,CDCl3):
0.83 (s,3H, CH3-19), 1.18 (s,3H, CH3-18),
1.23 (s, 3H, OCOCH3), 2.54 (s, 4H, NCH2), 5.01 (d, 1H, C17 H,
b-(benzo-1H-1,2,3-triazol-1-yl)-3b-piperidino-5a-andro-
d
b
24.40, 28.70, 31.03, 31.35, 31.87, 34.28, 36.00, 37.68, 37.80, 42.18,
45.65, 48.43, 50.01, 54.32, 58.18, 64.05, 67.29, 81.61, 145.06, 151.18,
170.99. Anal. Calcd. for C27H42N4O3: C 68.90, H 8.99, N 11.90;
Found C 68.72, H 8.85, N 11.99. MS (m/z):471[M þ 1]þ.
d
-a
J ¼ 9.08Hz), 5.28 (q,1H, C16
-aH), 7.33 (t,1H), 7.40e7.47 (m, 2H), 8.03
(d, 1H, J ¼ 8.28Hz). 13C NMR(150 MHz,CDCl3):
d 12.43, 12.72, 19.81,
20.43, 23.84, 24.64, 26.05, 28.72, 30.36, 31.38, 31.96, 34.42, 36.09,
37.84, 37.93, 42.79, 45.99, 48.75, 50.23, 54.40, 56.75, 64.58, 80.52,
109.98, 119.59, 123.73, 126.98, 134.09, 145.51, 170.58. Anal. Calcd. for
C32H46N4O2: C 74.09, H 8.94, N 10.80; Found C 74.13, H 8.80, N 10.91.
MS (m/z):519[M þ 1]þ.
5.1.6.11. 16
androstan-17
211e214 ꢀC; 1H NMR(400 MHz,CDCl3):
(s,3H, CH3-18), 1.72 (s, 3H, OCOCH3), 2.28 (s, 3H, NeCH3), 2.46e2.62
(br, 8H, NCH2), 3.72 (s, 4H, OCH2), 4.70 (d, 1H, C17
H, J ¼ 9.12Hz),
5.00 (q, 1H, C16
H), 7.90 (s, 1H), 7.92 (s, 1H). 13C
NMR(150 MHz,CDCl3): 12.36, 12.61, 20.14, 20.42, 24.46, 28.70,
b
-(1H-1,2,3-triazol-1-yl)-3
-yl acetate (17d). White solid. Yield 93%. m.p.
0.81 (s,3H, CH3-19), 1.12
b-(4-Methylpiperazinyl)-5a-
b
d
-a
-
a
5.1.6.17. 16
androstan-17
246e248 ꢀC; 1H NMR(400 MHz,CDCl3):
(s,3H, CH3-18), 1.23 (s, 3H, OCOCH3), 2.57 (s, 4H, NCH2), 3.73 (s, 4H,
OCH2), 5.02 (d, 1H, C17 H), 7.33 (t,
H, J ¼ 9.08Hz), 5.30 (q, 1H, C16
1H), 7.39e7.47 (m, 2H), 8.03 (d, 1H,
8.32Hz). 13C
b
-(benzo-1H-1,2,3-triazol-1-yl)-3
-yl acetate (18c). White solid. Yield 93%. m.p.
0.84 (s,3H, CH3-19), 1.19
b-morpholino-5a-
d
b
31.22, 31.34, 31.85, 34.26, 35.97, 37.79, 42.15, 45.76, 46.00, 48.42,
49.24, 54.31, 55.46, 58.14, 63.73, 81.60, 145.07, 151.19, 170.95. Anal.
Calcd. for C28H45N5O2: C 69.53, H 9.38, N 14.48; Found C 69.70, H
9.29, N 14.54. MS (m/z):484[M þ 1]þ.
d
-a
-a
J
¼

3672
H. Guo et al. / European Journal of Medicinal Chemistry 46 (2011) 3662e3674
NMR(150 MHz,CDCl3):
d
12.41, 12.73, 19.82, 20.44, 24.44, 28.76,
5.1.6.23. 16
17
-yl acetate (19b). White solid. Yield 71%. m.p. 256e258 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.82 (s,3H, CH3-19), 1.01 (s,3H, CH3-18),
b-(1H-benzimidazol-1-yl)-3b-piperidino-5a-androstan-
31.08, 31.39, 31.98, 34.44, 36.06, 37.74, 37.85, 42.80, 45.73, 48.77,
50.04, 54.42, 56.76, 64.10, 67.35, 80.53, 109.97, 119.62, 123.74,
127.00,134.10,145.53, 170.61. Anal. Calcd. for C31H44N4O3: C 71.51, H
8.52, N 10.76; Found C 71.44, H 8.39, N 10.80. MS (m/z):521[M þ 1]þ.
b
d
1.39 (s, 3H, OCOCH3), 2.57 (s, 4H, NCH2), 4.96e5.06 (m, 2H, C16-aH,
C17
-
a
H), 7.24e7.31 (m, 3H), 7.77 (d, 1H, J ¼ 6.84Hz), 8.05 (s, 1H). 13
C
NMR(150 MHz,CDCl3):
d 12.40, 13.51, 19.96, 20.45, 23.87, 24.71,
5.1.6.18. 16
-androstan-17
267e270 ꢀC; 1HNMR(400 MHz,CDCl3):
CH3-18), 1.23 (s, 3H, OCOCH3), 2.29 (s, 3H, NeCH3), 2.45e2.63 (br, 8H,
b
-(benzo-1H-1,2,3-triazol-1-yl)-3
-yl acetate (18d). White solid. Yield 91%. m.p.
0.84 (s,3H, CH3-19),1.19(s,3H,
b
-(4-Methylpiperazinyl)-
26.15, 28.66, 30.43, 31.98, 32.39, 34.31, 36.08, 37.71, 37.89, 42.93,
45.93, 48.62, 50.22, 53.88, 54.27, 64.41, 79.10, 109.93, 120.03,
122.13, 122.94, 134.78, 141.85, 142.99, 170.15. Anal. Calcd. for
C33H47N3O2: C 76.55, H 9.15, N 8.12; Found C 76.48, H 9.03, N 8.20.
MS (m/z):518[M þ 1]þ.
5a
b
d
NCH2), 5.01 (d,1H, C17
-
a
H, J ¼ 9.12Hz), 5.28 (q,1H, C16
-aH), 7.33 (t,1H),
7.39e7.47 (m, 2H), 8.03 (d, 1H, J ¼ 8.32Hz). 13C NMR(150 MHz,CDCl3):
d
12.41,12.71,19.81, 20.42, 24.36, 28.72, 31.12, 31.38, 31.96, 34.41, 36.02,
5.1.6.24. 16
17
-yl acetate (19c). White solid. Yield 80%. m.p. 235e238 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.83 (s,3H, CH3-19), 1.02 (s,3H, CH3-18),
1.39 (s, 3H, OCOCH3), 2.58 (s, 4H, NCH2), 3.73 (s, 4H, OCH2),
b-(1H-benzimidazol-1-yl)-3b-morpholino-5a-androstan-
37.78, 37.83, 42.78, 45.80, 45.83, 48.74, 49.04, 54.39, 55.23, 56.74, 63.75,
80.51,109.98,119.59,123.73,126.99,134.08,145.49,170.60. Anal. Calcd.
for C32H47N5O2: C 72.01, H 8.88, N 13.12; Found C 72.13, H 8.91, N 13.08.
MS (m/z):534[M þ 1]þ.
b
d
4.96e5.06 (m, 2H, C16-aH, C17-aH), 7.22e7.31 (m, 3H), 7.77 (d, 1H,
J ¼ 6.84Hz), 8.05 (s, 1H). 13C NMR(150 MHz,CDCl3):
d 12.37, 13.52,
5.1.6.19. 16
androstan-17
225e228 ꢀC; 1H NMR(400 MHz,CDCl3):
(s,3H, CH3-18), 1.23 (s, 3H, OCOCH3), 2.64 (q, 4H, NCH2), 5.02 (d, 1H,
b
-(benzo-1H-1,2,3-triazol-1-yl)-3
-yl acetate (18e). White solid. Yield 82%. m.p.
0.89 (s,3H, CH3-19), 1.19
b
-diethylamino-5
a
-
19.97, 20.45, 24.38, 28.66, 31.00, 31.97, 32.39, 34.31, 36.02, 37.64,
37.70, 42.93, 45.63, 48.62, 50.00, 53.88, 54.26, 64.03, 67.28, 79.10,
109.92, 120.02, 122.17, 122.96, 134.75, 141.83, 142.93, 170.18. Anal.
Calcd. for C32H45N3O3: C 73.95, H 8.73, N 8.09; Found C 73.88, H
8.67, N 8.14. MS (m/z):520[M þ 1]þ.
b
d
C17
-
a
H, J ¼ 9.08Hz), 5.30 (q, 1H, C16
-aH), 7.33 (t, 1H), 7.40e7.47 (m,
2H), 8.03 (d, 1H, J ¼ 8.36Hz). 13C NMR(150 MHz,CDCl3):
d. 12.41,
12.72, 13.70, 19.81, 20.43, 24.42, 28.75, 31.10, 31.38, 31.98, 34.42,
36.03, 37.80, 37.85, 42.80, 45.77, 48.77, 49.42, 54.42, 56.73, 64.02,
80.52, 109.97, 119.60, 123.73, 127.00, 134.09, 145.52, 170.60. Anal.
Calcd. for C31H46N4O2: C 73.48, H 9.15, N 11.06; Found C 73.52, H
9.03, N 11.18. MS (m/z):507[M þ 1]þ.
5.1.6.25. 16
androstan-17
288e290 ꢀC; 1H NMR(400 MHz,CDCl3):
(s,3H, CH3-18), 1.39 (s, 3H, OCOCH3), 2.28 (s, 3H, NeCH3), 2.40e2.61
b
-(1H-benzimidazol-1-yl)-3
-yl acetate (19d). White solid. Yield 80%. m.p.
0.82 (s,3H, CH3-19), 1.01
b-(4-Methylpiperazinyl)-5a-
b
d
(br, 8H, NCH2), 4.96e5.06 (m, 2H, C16-aH, C17-aH), 7.24e7.31 (m,
3H), 7.77 (d, 1H, J ¼ 6.92Hz), 8.05 (s, 1H). 13C NMR(150 MHz,CDCl3):
5.1.6.20. 16
-androstan-17
277e280 ꢀC; 1H NMR(400 MHz,CDCl3):
CH3-18), 1.24 (s, 3H, OCOCH3), 2.79 (s, 3H, NeCH3), 5.03 (d, 1H, C17-
b
-(benzo-1H-1,2,3-triazol-1-yl)-3
-yl acetate (18f). White solid. Yield 91%. m.p.
0.91 (s,3H, CH3-19),1.20 (s,3H,
H,
b
-(N-methylphenylamino)-
d 12.39, 13.51, 19.97, 20.43, 24.46, 28.65, 31.21, 31.97, 32.37, 34.29,
5a
b
36.00, 37.69, 37.74, 42.91, 45.73, 46.02, 48.60, 49.27, 53.85, 54.26,
55.48, 63.74, 79.08, 109.91, 120.03, 122.12, 122.92, 134.75, 141.83,
142.99, 170.16. Anal. Calcd. for C33H48N4O2: C 74.40, H 9.08, N 10.52;
Found C 74.29, H 9.16, N 10.44. MS (m/z):533[M þ 1]þ.
d
a
J ¼ 9.08Hz), 5.30 (q, 1H, C16- H), 6.68 (t, 1H), 6.79 (d, 2H, J ¼ 8.20Hz),
a
7.23 (t, 2H), 7.33 (t, 2H), 7.40e7.47 (m, 2H), 8.03 (d, 1H, J ¼ 8.32Hz). 13C
NMR(150 MHz,CDCl3):
d
12.53, 12.76, 19.84, 20.50, 25.01, 28.59, 29.71,
5.1.6.26. 16
stan-17
-yl acetate (19e). White solid. Yield 90%. m.p. 202e204 ꢀC;
1H NMR(400 MHz,CDCl3):
0.82 (s,3H, CH3-19), 1.01 (s,3H, CH3-18),
b-(1H-benzimidazol-1-yl)-3b-diethylamino-5a-andro-
31.39, 31.62, 31.95, 34.47, 35.93, 37.87, 37.98, 42.82, 46.25, 48.78, 54.43,
56.77, 58.33, 80.54, 110.00, 113.24, 116.34, 119.64, 123.76, 127.02, 129.12,
134.12, 145.55, 150.23, 170.62. Anal. Calcd. for C34H44N4O2: C 75.52, H
8.20, N 10.36; Found C 75.40, H 8.09, N 10.51. MS (m/z):541[M þ 1]þ.
b
d
1.40 (s, 3H, OCOCH3), 2.64 (s, 4H, NCH2), 4.96e5.06 (m, 2H, C16-aH,
C17
-
a
H), 7.24e7.31 (m, 3H), 7.77 (d, 1H, J ¼ 7.00Hz), 8.05 (s, 1H). 13
C
NMR(150 MHz,CDCl3): d.12.38,13.50,13.71,19.98, 20.45, 24.38, 28.64,
5.1.6.21. 16
-androstan-17
198e201 ꢀC; 1H NMR(400 MHz,CDCl3):
CH3-18), 1.23 (s, 3H, OCOCH3), 2.22 (s, 3H, NeCH3), 3.59 (s, 2H, PhCH2),
b
-(benzo-1H-1,2,3-triazol-1-yl)-3
-yl acetate (18g). White solid. Yield 79%. m.p.
0.86 (s,3H, CH3-19), 1.19 (s,3H,
b
-(N-benzylmethylamino)-
31.14, 31.97, 32.38, 34.25, 36.01, 37.68, 37.70, 42.91, 45.71, 48.62, 49.33,
53.88, 54.25, 64.02, 79.10, 109.92, 120.02, 122.16, 122.94, 134.75,
141.83, 142.97, 170.16. Anal. Calcd. for C32H47N3O2: C 76.00, H 9.37, N
8.31; Found C 76.12, H 9.29, N 8.40. MS (m/z):506[M þ 1]þ.
5a
b
d
5.01 (d, 1H, C17-
a
H, J ¼ 9.04Hz), 5.29 (q, 1H, C16-
aH), 7.23 (t, 2H),
7.29e7.35 (m, 4H), 7.40e7.47 (m, 2H), 8.03 (d, 1H, J ¼ 8.32Hz). 13C
5.1.6.27. 16
androstan-17
240 ꢀC; 1H NMR(400 MHz,CDCl3):
18), 1.40 (s, 3H, OCOCH3), 2.79 (s, 3H, NeCH3), 4.96e5.06 (m, 2H,
b
-(1H-benzimidazol-1-yl)-3
-yl acetate (19f). White solid. Yield 79%. m.p. 236e
0.89 (s,3H, CH3-19), 1.03 (s,3H, CH3-
b-(N-methylphenylamino)-5a-
NMR(150 MHz,CDCl3):
d
12.47, 12.75, 19.85, 20.48, 23.96, 28.80, 30.47,
b
31.42, 32.00, 34.46, 36.18, 37.83, 37.89, 37.93, 42.81, 45.98, 48.79, 54.44,
56.77, 58.01, 62.62, 80.55, 110.00, 119.63, 123.74, 126.75, 127.00, 128.21,
128.86, 134.11, 145.55, 170.63. Anal. Calcd. for C35H46N4O2: C 75.78, H
8.36, N 10.10; Found C 75.61, H 8.50, N 10.22. MS (m/z):555[M þ 1]þ.
d
C16-
a
H, C17-
a
H), 6.69 (t, 1H), 6.79 (d, 2H, J ¼ 8.12Hz), 7.20e7.30 (m, 5H),
7.77 (d, 1H, J ¼ 6.92Hz), 8.06 (s, 1H). 13C NMR(150 MHz,CDCl3):
d 12.51,
13.56, 20.01, 20.52, 24.98, 28.50, 29.71, 31.41, 31.58, 31.95, 32.42, 34.35,
35.90, 37.73, 37.91, 42.94, 46.16, 48.63, 53.89, 54.27, 58.29, 79.10, 109.95,
113.27, 116.41, 120.07, 122.18, 122.99, 129.13, 141.85, 150.20, 170.20. Anal.
Calcd. for C35H45N3O2: C 77.88, H 8.40, N 7.79; Found C 77.91, H 8.32, N
7.87. MS (m/z):540[M þ 1]þ.
5.1.6.22. 16
17
-yl acetate (19a). White solid. Yield 77%. m.p. 241e243 ꢀC; 1H
NMR(400 MHz,CDCl3): 0.86 (s,3H, CH3-19), 1.01 (s,3H, CH3-18),
b-(1H-benzimidazol-1-yl)-3b-pyrrolidino-5a-androstan-
b
d
1.39 (s, 3H, OCOCH3), 2.87 (s, 4H, NCH2), 4.96e5.06 (m, 2H, C16-aH,
C17
-
a
H), 7.22e7.31 (m, 3H), 7.77 (d, 1H, J ¼ 6.96Hz), 8.05 (s, 1H). 13
C
NMR(150 MHz,CDCl3):
d
12.20, 13.48, 19.93, 20.38, 25.99, 28.38,
5.1.6.28. 16
androstan-17
203 ꢀC; 1H NMR(400 MHz,CDCl3):
CH3-18), 1.41 (s, 3H, OCOCH3), 2.29 (s, 3H, NeCH3), 3.72 (s, 2H, PhCH2),
b
-(1H-benzimidazol-1-yl)-3
-yl acetate (19g). White solid. Yield 72%. m.p. 201e
0.84 (s,3H, CH3-19), 1.00 (s,3H,
b-(N-benzylmethylamino)-5a-
31.83, 32.30, 32.60, 34.20, 35.77, 37.07, 37.62, 42.88, 45.11, 48.54,
50.97, 51.62, 53.84, 54.05, 63.88, 79.03, 119.95, 122.13, 122.92,
134.71, 141.81, 142.87, 170.13. Anal. Calcd. for C32H45N3O2: C 76.30,
H 9.00, N 8.34; Found C 76.09, H 8.91, N 8.40. MS (m/z):504
[M þ 1]þ.
b
d
4.96e5.06 (m, 2H, C16-
a
H, C17-
a
H), 6.70 (t, 1H), 6.79 (d, 2H, J ¼ 8.14Hz),
7.23 (t, 2H), 7.21e7.31 (m, 4H), 7.77 (d, 1H, J ¼ 6.96Hz), 8.05 (s, 1H). 13C

H. Guo et al. / European Journal of Medicinal Chemistry 46 (2011) 3662e3674
3673
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NMR(150 MHz,CDCl3):
d 12.41, 13.74, 17.12, 19.18, 20.19, 23.54, 28.68,
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5.2.2. Cell viability assay(MTT)
The anticancer activity in vitro was measured using the MTT
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20
m
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mL DMSO
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mM, were obtained by a linear regression analysis of
the concentration-response curves obtained for each compound.
Assays were performed in triplicate on three independent
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This work was financially supported by the Important National
Science & Technology Specific Projects (No. 2009ZX09301-014-1)
and the Fundamental Research Funds for the Central Universities.
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