Electrophile-induced generation of cyclic azomethine imines from steroidal δ-alkenyl hydrazones

Article abstract of DOI:10.1016/j.steroids.2009.01.001

Novel δ-alkenyl phenylhydrazones were synthesized in both the estrone and the 13α-estrone series. Electrophile-induced cyclizations of alkenyl phenylhydrazones with phenylselenyl bromide furnished cyclic iminium salts, via attack of the imino nitrogen ato

Full text of DOI:10.1016/j.steroids.2009.01.001

Steroids 74 (2009) 474–482
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Steroids
journal homepage: www.elsevier.com/locate/steroids
Electrophile-induced generation of cyclic azomethine imines from steroidal
␦-alkenyl hydrazones
Erzsébet Mernyáka,∗, László Márkb, Éva Franka, Gyula Schneidera, János Wölflinga
a Department of Organic Chemistry, University of Szeged, Dóm tér 8, H-6720 Szeged, Hungary
b Department of Biochemistry and Medical Chemistry, University of Pécs, Szigeti út 12, H-7624 Pécs, Hungary
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 8 August 2008
Received in revised form 19 December 2008
Accepted 5 January 2009
Available online 13 January 2009
Novel ␦-alkenyl phenylhydrazones were synthesized in both the estrone and the 13␣-estrone series.
Electrophile-induced cyclizations of alkenyl phenylhydrazones with phenylselenyl bromide furnished
cyclic iminium salts, via attack of the imino nitrogen atom on the intermediate seleniranium ion. Hydride
reduction of the iminium salts led to novel aminophenyl-substituted aza-D-homoestrones. The structures
of the new products were determined by NMR (one- and two-dimensional) and MALDI-MS techniques,
with C70 fullerenes as matrix in the latter case.
Keywords:
© 2009 Elsevier Inc. All rights reserved.
Phenylhydrazones
Stereoselectivity
Steroids
Phenylselenyl bromide
Aza-D-homoestrone
C70 fullerenes
1. Introduction
Lewis-acid induced formation and subsequent cyclization of
steroidal azomethine imines [18].
Cyclofunctionalizations based on the reaction of an electrophilic
reagent with an alkene possessing a suitably positioned nucle-
ophilic group are unchangedly receiving attention. Organoselenium
reagents are often used as electrophiles, owing to their ready
availability, the mild reaction conditions, and the numerous chem-
ical modifications which can be performed on the selenium
moiety. Amines, imidates, imines, azides, and amino acids, for
instance, have been used as internal N nucleophiles in selenocy-
clizations of alkenes [1–4]. The phenylselenyl bromide-induced
cyclization of alkenyl oximes or oxime ethers leads to the seleni-
ranium ion, which can be attacked by the O or the N atom of
the ambident nucleophile [5–9]. Our previous work revealed the
nucleophilic attack of the oxime O or N atom of steroidal sec-
ooximes or oxime ethers, yielding oxazepines and cyclic nitrones,
which underwent intermolecular 1,3-dipolar cycloaddition with
each other [9]. The resulting non-symmetrical steroid dimers
contained 16-halomethyl- or phenylselenylmethyl-substituted six-
and seven-membered D rings.
Our present goal was to synthesize D-secophenylhydrazones
in both the estrone and the 13␣-estrone series. Additionally, the
phenylselenyl bromide-induced cyclizations of the hydrazones,
yielding cyclic azomethine imine salts, are discussed.
2. Experimental
Melting points (mps) were determined with a Kofler hot-
stage apparatus and are uncorrected. Elemental analyses were
performed with a PerkinElmer CHN analyzer model 2400. Thin-
layer chromatography: silica gel 60 F254; layer thickness 0.2 mm
(Merck); detection with iodine or UV (365 nm) after spraying
with 5% phosphomolybdic acid in 50% aqueous phosphoric acid
and heating at 100–120 ◦C for 10 min. Flash chromatography: sil-
ica gel 60, 40–63 ␮m (Merck). 1H NMR spectra were recorded
in CDCl3 solution (if not otherwise stated) with a Bruker DRX-
500 instrument at 500 MHz, with Me4Si as internal standard.
13C NMR spectra were recorded with the same instrument at
125 MHz under the same conditions. The mass spectrometer
used was an Autoflex II TOF/TOF (Bruker Daltonics, Bremen, Ger-
many) operated in reflector mode. The ions were accelerated
under delayed extraction conditions (80 ns) in positive and neg-
ative ion modes, with an acceleration voltage of 20.00 kV. The
instrument uses a 337 nm pulsed (50 Hz) nitrogen laser. 1 ␮L
aliquots of the standard solutions were loaded onto the target
Azomethine imines are usually generated by thermal,
microwave-, or acid-induced 1,2-prototropy from the termi-
nal N to the central N atom [10–17]. We recently described the
∗
Corresponding author. Fax: +36 62 544199.
E-mail address: bobe@chem.u-szeged.hu (E. Mernyák).
0039-128X/$ – see front matter © 2009 Elsevier Inc. All rights reserved.
doi:10.1016/j.steroids.2009.01.001

E. Mernyák et al. / Steroids 74 (2009) 474–482
475
plate (MTP 384 target plate ground steel TF, Bruker Daltonics,
Bremen, Germany) by mixing with the same volume of a sat-
urated matrix solution prepared by dissolving C70 fullerenes in
toluene. EI-MS spectra were measured on a Varian MAT 311A instru-
ment.
to react. After work-up, 3c was obtained as a white solid (400 mg,
82%). mp 173–175 ◦C; Rf = 0.18 (60% CH2Cl2/40% hexane). Anal.
Calcd. for C33H35N3O: C, 80.95; H, 7.20. Found: C, 81.07; H, 7.05.
1H NMR (CDCl3): ı = 1.16 (s, 3H, 18-H3), 2.83 (m, 2H, 6-H2), 4.94
(m, 2H, 16a-H2), 5.03 (s, 2H, OCH2), 5.83 (m, 1H, 16-H), 6.72 (d,
1H, J = 2.0 Hz, 4-H), 6.79 (dd, 1H, J = 8.6 Hz, J = 2.0 Hz, 2-H), 6.93
(s, 1H, 17-H), 6.98 (d, 2H, J = 8.3 Hz, 2ꢀ-H and 6ꢀ-H), 7.19 (d, 1H,
J = 8.6 Hz, 1-H), 7.31 (t, 1H, J = 7.1 Hz, 4ꢀꢀ-H), 7.37 (t, 2H, J = 7.1 Hz,
3ꢀꢀ-H and 5ꢀꢀ-H), 7.42 (d, 2H, J = 7.1 Hz, 2ꢀꢀ-H and 6ꢀꢀ-H), 7.47 (d,
2H, J = 8.3 Hz, 3ꢀ-H and 5ꢀ-H), 7.54 (s, 1H, NH) ppm. 13C NMR
(CDCl3): ı = 16.1 (C-18), 26.0, 27.4, 30.3, 34.2, 38.0, 41.0, 42.5
(C-13), 43.3, 47.7, 70.0 (OCH2), 101.0 (C-4ꢀ), 112.2 (2C, C-2ꢀ and
C-6ꢀ), 112.5 (C-2), 114.5 (C-4), 114.7 (C-16a), 120.1 (CN), 126.4 and
127.4 (2C) and 127.8 and 128.5 (2C): C-1, C-2ꢀꢀ, C-3ꢀꢀ, C-4ꢀꢀ, C-5ꢀꢀ
and C-6ꢀꢀ, 132.5 (C-10), 133.6 (2C, C-3ꢀ and C-5ꢀ), 137.2 (C-1ꢀꢀ),
137.9 (C-5), 139.8 (C-16), 148.5 (C-1ꢀ), 152.5 (C-17), 156.8 (C-3).
MS negative mode: 488 (10% [M−H]−), 117 (100% [NHPhCN]−);
positive mode: 489 (10% [M]+), 270 (100%), 91 (30%, benzylic
cation).
2.1. General procedure for the synthesis of steroidal
phenylhydrazones
Secoaldehyde 1 or 2 [19] (375 mg, 1.00 mmol) was dissolved
in methanol (5 mL), and phenyl- or substituted phenylhydrazine
hydrochloride (1.00 mmol) and a methanolic solution (5 mL) of
sodium acetate (200 mg, 2.44 mmol) were added. The mixture was
stirred for 0.5 h at rt. The white or yellow precipitate was filtered
off, washed with cold methanol, and dried.
2.2. Reaction of 1 with phenylhydrazine hydrochloride
According to Section 2.1, 1 (375 mg, 1.00 mmol), phenylhy-
drazine hydrochloride (145 mg, 1.00 mmol) and sodium acetate
(200 mg, 2.44 mmol) in methanol were allowed to react. After work-
up, 3a was obtained as a white solid (395 mg, 85%). mp 137–139 ◦C;
Rf = 0.54 (60% CH2Cl2/40% hexane). Anal. Calcd. for C32H36N2O: C,
82.72; H, 7.81. Found: C, 82.90; H, 8.02. 1H NMR (CDCl3): ı = 1.16 (s,
3H, 18-H3), 2.83 (m, 2H, 6-H2), 4.93 (m, 2H, 16a-H2), 5.03 (s, 2H,
OCH2), 5.84 (m, 1H, 16-H), 6.71 (d, 1H, J = 2.4 Hz, 4-H), 6.77–6.82
(overlapping multiplets, 2H, 2-H and 4ꢀ-H), 6.86 (s, 1H, 17-H), 6.99
(d, 2H, J = 8.1 Hz, 2ꢀ-H and 6ꢀ-H), 7.13 (s, 1H, NH), 7.19–7.24 (overlap-
ping multiplets, 3H, 1-H, 3ꢀ-H and 5ꢀ-H), 7.30 (t, 1H, J = 7.3 Hz, 4ꢀꢀ-H),
7.37 (t, 2H, J = 7.3 Hz, 3ꢀꢀ-H and 5ꢀꢀ-H), 7.42 (d, 2H, J = 7.3 Hz, 2ꢀꢀ-H and
6ꢀꢀ-H) ppm. 13C NMR (CDCl3): ı = 16.2 (C-18), 26.1, 27.5, 30.3, 34.2,
38.2, 41.1, 42.3 (C-13), 43.4, 47.8, 70.0 (OCH2), 112.5 (3C, C-2, C-2ꢀ
and C-6ꢀ), 114.5 (C-16a), 114.6 (C-4), 119.4 (C-4ꢀ), 126.4 (C-1), 127.4
(2C) and 128.5 (2C): C-2ꢀꢀ, C-3ꢀꢀ, C-5ꢀꢀ and C-6ꢀꢀ, 127.8 (C-4ꢀꢀ), 129.2
(2C, C-3ꢀ and C-5ꢀ), 132.8 (C-10), 137.3 (C-1ꢀꢀ), 138.0 (C-5), 140.1 (C-
16), 145.6 (C-1ꢀ), 149.7 (C-17), 156.8 (C-3). MS positive mode: 463
(20% [M−H]+), 91 (100%, benzylic cation).
2.5. Reaction of 1 with 4-nitrophenylhydrazine hydrochloride
According to Section 2.1,
1
(375 mg, 1.00 mmol), 4-
nitrophenylhydrazine hydrochloride (190 mg, 1.00 mmol) and
sodium acetate (200 mg, 2.44 mmol) in methanol were allowed to
react. After work-up, 3d was obtained as a yellow solid (510 mg,
90%). mp 186–188 ◦C; Rf = 0.25 (60% CH2Cl2/40% hexane). Anal.
Calcd. for C32H35N3O3: C, 75.41; H, 6.92. Found: C, 75.27; H, 7.17.
1H NMR (CDCl3): ı = 1.18 (s, 3H, 18-H3), 2.84 (m, 2H, 6-H2), 4.95
(m, 2H, 16a-H2), 5.04 (s, 2H, OCH2), 5.82 (m, 1H, 16-H), 6.72 (d,
1H, J = 2.4 Hz, 4-H), 6.79 (dd, 1H, J = 8.5 Hz, J = 2.4 Hz, 2-H), 6.98
(overlapping multiplets, 3H, 2ꢀ-H, 6ꢀ-H and 17-H), 7.20 (d, 1H,
J = 8.5 Hz, 1-H), 7.31 (t, 1H, J = 7.2 Hz, 4ꢀꢀ-H), 7.38 (t, 2H, J = 7.2 Hz,
3ꢀꢀ-H and 5ꢀꢀ-H), 7.42 (d, 2H, J = 7.2 Hz, 2ꢀꢀ-H and 6ꢀꢀ-H). 7.73 (s, 1H,
NH), 8.13 (d, 2H, J = 8.3 Hz, 3ꢀ-H and 5ꢀ-H) ppm. 13C NMR (CDCl3):
ı = 16.1 (C-18), 26.0, 27.4, 30.3, 34.2, 38.0, 41.0, 42.6 (C-13), 43.3,
47.6, 70.0 (OCH2), 111.3 (2C, C-2ꢀ and C-6ꢀ), 112.5 (C-2), 114.5 (C-4),
114.7 (C-16a), 126.2 (2C, C-3ꢀ and C-5ꢀ), 126.4 and 127.4 (2C) and
127.9 and 128.5 (2C): C-1, C-2ꢀꢀ, C-3ꢀꢀ, C-4ꢀꢀ, C-5ꢀꢀ and C-6ꢀꢀ, 132.5
(C-10), 137.3 (C-1ꢀꢀ), 137.9 (C-5), 139.7 (C-16), 139.8 (C-4ꢀ), 150.2
(C-1ꢀ), 153.7 (C-17), 156.8 (C-3). MS negative mode: 508 (60%
[M−H]−); positive mode: 508 (40% [M−H]+); 91 (100%, benzylic
cation).
2.3. Reaction of 1 with 4-tolylhydrazine hydrochloride
According to Section 2.1,
1
(375 mg, 1.00 mmol), 4-
tolylhydrazine hydrochloride (159 mg, 1.00 mmol) and sodium
acetate (200 mg, 2.44 mmol) in methanol were allowed to react.
After work-up, 3b was obtained as a white solid (417 mg, 87%). mp
149–152 ◦C; Rf = 0.57 (60% CH2Cl2/40% hexane). Anal. Calcd. for
33H38N2O: C, 82.80; H, 8.00. Found: C, 82.53; H, 7.92. 1H NMR
2.6. Reaction of 2 with phenylhydrazine hydrochloride
C
(CDCl3): ı = 1.15 (s, 3H, 18-H3), 2.26 (s, 3H, tolyl-CH3), 2.83 (m, 2H,
6-H2), 4.92 (m, 2H, 16a-H2), 5.02 (s, 2H, OCH2), 5.84 (m, 1H, 16-H),
6.71 (d, 1H, J = 2.0 Hz, 4-H), 6.78 (dd, 1H, J = 8.6 Hz, J = 2.0 Hz, 2-H),
6.84 (s, 1H, 17-H), 6.90 (d, 2H, J = 8.3 Hz, 2ꢀ-H and 6ꢀ-H), 7.02 (d,
2H, J = 8.3 Hz, 3ꢀ-H and 5ꢀ-H), 7.20 (d, 1H, J = 8.6 Hz, 1-H), 7.30 (t,
1H, J = 7.1 Hz, 4ꢀꢀ-H), 7.37 (t, 2H, J = 7.1 Hz, 3ꢀꢀ-H and 5ꢀꢀ-H), 7.42 (d,
2H, J = 7.1 Hz, 2ꢀꢀ-H and 6ꢀꢀ-H) ppm. 13C NMR (CDCl3): ı = 16.2, 20.5,
26.1, 27.5, 30.3, 34.2, 38.3, 41.1, 42.2, 43.4, 47.8, 70.0 (OCH2), 112.5
(C-2), 112.7 (2C, C-2ꢀ and C-6ꢀ), 114.4 (C-16a), 114.5 (C-4), 126.4
(C-1), 127.4 (2C) and 128.5 (2C): C-2ꢀꢀ, C-3ꢀꢀ, C-5ꢀꢀ and C-6ꢀꢀ, 127.8
(C-4ꢀꢀ), 128.6 (C-4ꢀ), 129.6 (2C, C-3ꢀ and C-5ꢀ), 132.8 (C-10), 137.4
(C-1ꢀ), 138.0 (C-5), 140.1 (C-16), 143.5 (C-1ꢀ), 149.3 (C-17), 156.8
(C-3). MS positive mode: 477 (20% [M−H]+), 91 (100%, benzylic
cation).
According to Section 2.1, 2 (375 mg, 1.00 mmol), phenylhy-
drazine hydrochloride (145 mg, 1.00 mmol) and sodium acetate
(200 mg, 2.44 mmol) in methanol were allowed to react. After work-
up, 4a was obtained as a white solid (465 mg, 86%). mp 100–102 ◦C;
Rf = 0.60 (60%CH2Cl2/40% hexane). Anal. Calcd. for C32H36N2O: C,
82.72; H, 7.81. Found: C, 82.65; H, 7.98. 1H NMR (CDCl3): ı = 1.27 (s,
3H, 18-H3), 2.85 (m, 2H, 6-H2), 4.99 (m, 2H, 16a-H2), 5.03 (s, 2H,
OCH2), 5.87 (m, 1H, 16-H), 6.71 (d, 1H, J = 2.2 Hz, 4-H), 6.77–6.82
(overlapping multiplets, 2H, 2-H and 4ꢀ-H), 6.95 (d, 2H, J = 7.9 Hz,
2ꢀ-H and 6ꢀ-H), 7.12 (s, 1H, NH), 7.18–7.23 (overlapping multiplets,
4H, 1-H and 17-H and 3ꢀ-H and 5ꢀ-H), 7.31 (t, 1H, J = 7.1 Hz, 4ꢀꢀ-H),
7.38 (t, 2H, J = 7.1 Hz, 3ꢀꢀ-H and 5ꢀꢀ-H), 7.42 (d, 2H, J = 7.1 Hz, 2ꢀꢀ-H and
6ꢀꢀ-H) ppm. 13C NMR (CDCl3): ı = 27.1 (C-18), 27.2, 27.5, 30.4, 33.4,
39.5, 41.6, 42.4, 43.6, 51.1, 70.0 (OCH2), 112.5 (3C, C-2, C-2ꢀ and C-
6ꢀ), 114.5 (C-4), 114.7 (C-16a), 119.4 (C-4ꢀ), 126.5 and 127.4 (2C) and
127.8 and 128.5 (2C): C-1, C-2ꢀꢀ, C-3ꢀꢀ, C-4ꢀꢀ, C-5ꢀꢀ and C-6ꢀꢀ, 129.2 (2C,
C-3ꢀ and C-5ꢀ), 132.9 (C-10), 137.3 (C-1ꢀꢀ), 137.9 (C-5), 139.9 (C-16),
144.7 (C-17), 145.5 (C-1ꢀ), 156.8 (C-3). MS positive mode: 464 (20%
[M]+), 91 (100%, benzylic cation).
2.4. Reaction of 1 with 4-cyanophenylhydrazine hydrochloride
According to Section 2.1,
1
(375 mg, 1.00 mmol), 4-
cyanophenylhydrazine hydrochloride (170 mg, 1.00 mmol) and
sodium acetate (200 mg, 2.44 mmol) in methanol were allowed

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E. Mernyák et al. / Steroids 74 (2009) 474–482
2.7. Reaction of 2 with 4-tolylhydrazine hydrochloride
C-6ꢀꢀ), 127.8 (C-1), 128.5 (2C, C-3ꢀꢀ and C-5ꢀꢀ), 132.7 (C-10), 137.2
(C-1ꢀꢀ), 137.8 (C-5), 139.3 (C-16), 139.7 (C-4ꢀ), 149.3 (C-17), 150.1
(C-1ꢀ), 156.8 (C-3). MS negative mode: 508 (80% [M−H]−); positive
mode: 508 (40% [M−H]+), 91 (100%, benzylic cation).
According to Section 2.1,
2
(375 mg, 1.00 mmol), 4-
tolylhydrazine hydrochloride (159 mg, 1.00 mmol) and sodium
acetate (200 mg, 2.44 mmol) in methanol were allowed to react.
After stirring for 30 min, the reaction mixture was cooled to
−18 ◦C, when a light-yellow precipitate of 4b appeared (480 mg,
92%). mp 86–88 ◦C; Rf = (50% CH2Cl2/50% hexane). Anal. Calcd. for
2.10. General procedure for the synthesis of
16-phenylselenylmethyl-17aza-D-homoestrones
C
33H38N2O: C, 82.80; H, 8.00. Found: C, 82.76; H, 7.84.
Hydrazone 3 (1.00 mmol) or 4 (1.00 mmol) was dissolved
in dry acetonitrile (10 mL), and phenylselenyl bromide (236 mg,
1.00 mmol) was added in an ice-water bath under a nitrogen
atmosphere. The mixture was stirred for 0.5 h, the solvent was evap-
orated off, the crude product was dissolved in dichloromethane
(5 mL), methanol (5 mL) was added, and the reduction was
carried out with potassium borohydride (216 mg, 4 mmol) dur-
ing 24 h at room temperature under stirring. The mixture was
diluted with dichloromethane (30 mL) and washed with water. The
dichloromethane solution was dried over anhydrous sodium sul-
fate and evaporated under reduced pressure. The resulting oil was
purified by flash chromatography.
1H NMR (CDCl3): ı = 1.28 (s, 3H, 18-H3), 2.27 (s, 3H, tolyl-CH3),
2.84 (m, 2H, 6-H2), 5.00 (m, 2H, 16a-H2), 5.04 (s, 2H, OCH2), 5.87
(m, 1H, 16-H), 6.72 (d, 1H, J = 2.3 Hz, 4-H), 6.80 (dd, 1H, J = 8.4 Hz,
J = 2.3 Hz, 2-H), 6.88 (d, 2H, J = 8.3 Hz, 2ꢀ-H and 6ꢀ-H), 7.03 (d, 2H,
J = 8.3 Hz, 3ꢀ-H and 5ꢀ-H), 7.18 (s, 1H, 17-H), 7.21 (d, 1H, J = 8.4 Hz, 1-
H), 7.32 (t, 1H, J = 7.1 Hz, 4ꢀꢀ-H), 7.39 (t, 2H, J = 7.1 Hz, 3ꢀꢀ-H and 5ꢀꢀ-H),
7.44 (d, 2H, J = 7.1 Hz, 2ꢀꢀ-H and 6ꢀꢀ-H) ppm. 13C NMR (CDCl3): ı = 20.5
(tolyl-CH3), 27.1 (C-18), 27.3, 27.6, 30.4, 33.4, 39.6, 41.6 (C-13), 42.4,
43.7, 51.2, 70.0 (OCH2), 112.5 (C-2), 112.6 (2C, C-2ꢀ and C-6ꢀ), 114.5
(C-4), 114.6 (C-16a), 126.5 and 127.8: C-1 and C-4ꢀꢀ, 127.4 (2C) and
128.5 (2C) and 129.6 (2C): C-2ꢀꢀ, C-3ꢀꢀ, C-5ꢀꢀ, C-6ꢀꢀ, C-3ꢀ and C-5ꢀ), 128.7
(C-4ꢀ), 133.0 (C-10), 137.4 (C-1ꢀꢀ), 137.9 (C-5), 140.0 (C-16), 143.4 (C-
1ꢀ), 144.4 (C-17), 156.8 (C-3). MS positive mode: 477 (30% [M−H]+),
91 (100%, benzylic cation).
2.11. Cyclization of 3a
According to Section 2.10, hydrazone 3a (502 mg, 1.00 mmol)
and phenylselenyl bromide (236 mg, 1.00 mmol) were allowed to
react in acetonitrile (10 mL). After reduction, the crude product was
subjected to chromatographic purification on a silica gel column
with light petroleum/dichloromethane (50:50) as eluent, but 15
remained as an oil (430 mg, 69%). Rf = 0.40 (50% CH2Cl2/50% hex-
ane). Anal. Calcd. for C38H42N2OSe: C, 73.41; H, 6.81. Found: C,
73.62; H, 6.95. 1H NMR (CDCl3): ı = 1.12 (s, 3H, 18-H3), 1.94 (d, 1H,
J = 10.1 Hz) and 2.89 (d, 1H, J = 10.1 Hz): 17a-H2, 2.58 (m, 1H, 16-H),
2.84 (m, 2H, 6-H2), 3.14 (dd, 1H, J = 11.9 Hz, J = 7.3 Hz) and 3.41 (dd,
1H, J = 11.9 Hz, J = 2.9 Hz): 16a-H2, 4.04 (s, 1H, NH), 5.04 (s, 2H, OCH2),
6.72 (d, 1H, J = 2.6 Hz, 4-H), 6.78 (overlapping multiplets, 2H, 2-H
and 4ꢀ-H), 6.91 (d, 2H, J = 7.6 Hz, 2ꢀ-H and 6ꢀ-H), 7.18–7.22 (overlap-
ping multiplets, 6H, aromatic protons), 7.31 (t, 1H, J = 7.1 Hz, 4ꢀꢀ-H),
7.38 (t, 2H, J = 7.1 Hz, 3ꢀꢀ-H and 5ꢀꢀ-H), 7.43 (d, 2H, J = 7.1 Hz, 2ꢀꢀ-H and
6ꢀꢀ-H), 7.47 (m, 2H) ppm. 13C NMR (CDCl3): ı = 17.6 (C-18), 25.7, 26.2,
30.0, 31.1, 32.8, 35.4, 37.7, 38.6, 43.6, 47.6, 66.3 (C-16), 70.0 (OCH2),
70.4 (NCH2), 112.4 (C-2), 113.5 (2C, C-2ꢀ and C-6ꢀ), 114.6 (C-4), 119.2
(C-4ꢀ), 126.1, 126.3, 127.4 (2C), 127.8, 128.5 (2C), 128.9 (2C), 129.1
(2C), 131.9 (3C), 133.0 (C-10), 137.3 (C-1ꢀꢀ), 137.9 (C-5), 148.3 (C-1ꢀ),
156.8 (C-3). MS (EI); m/z (%): 622 (100, M+).
2.8. Reaction of 2 with 4-cyanophenylhydrazine hydrochloride
According to Section 2.1,
2
(375 mg, 1.00 mmol), 4-
cyanophenylhydrazine hydrochloride (170 mg, 1.00 mmol) and
sodium acetate (200 mg, 2.44 mmol) in methanol were allowed
to react. After work-up, 4c was obtained as a white solid (392 mg,
80%). mp 135–136 ◦C; Rf = 0.21 (60% CH2Cl2/40% hexane). Anal.
Calcd. for C33H35N3O: C, 80.95; H, 7.20. Found: C, 81.11; H, 7.36. 1
H
NMR (CDCl3): ı = 1.23 (s, 3H, 18-H3), 2.79 (m, 2H, 6-H2), 4.98 (m,
2H, 16a-H2), 5.00 (s, 2H, OCH2), 5.80 (m, 1H, 16-H), 6.67 (d, 1H,
J = 2.0 Hz, 4-H), 6.73 (dd, 1H, J = 8.6 Hz, J = 2.0 Hz, 2-H), 6.91 (d, 2H,
J = 8.3 Hz, 2ꢀ-H and 6ꢀ-H), 7.15 (d, 1H, J = 8.6 Hz, 1-H), 7.20 and 7.21
(2×s, 2×1H, 17-H and NH), 7.27 (t, 1H, J = 7.1 Hz, 4ꢀꢀ-H), 7.33 (t, 2H,
J = 7.1 Hz, 3ꢀꢀ-H and 5ꢀꢀ-H), 7.37 (d, 2H, J = 7.1 Hz, 2ꢀꢀ-H and 6ꢀꢀ-H), 7.41
(d, 2H, J = 8.3 Hz, 3ꢀ-H and 5ꢀ-H) ppm. 13C NMR (CDCl3): ı = 26.9
(C-18), 27.2, 27.4, 30.3, 33.3, 39.3, 41.9 (C-13), 42.3, 43.5, 50.9, 70.0
(OCH2), 100.9 (C-4ꢀ), 112.1 (2C, C-2ꢀ and C-6ꢀ), 112.5 (C-2), 114.5
(C-4), 115.0 (C-16a), 120.1 (CN), 126.4 and 127.4 (2C) and 127.8
and 128.5 (2C): C-1, C-2ꢀꢀ, C-3ꢀꢀ, C-4ꢀꢀ, C-5ꢀꢀ and C-6ꢀꢀ, 132.7 (C-10),
133.6 (2C, C-3ꢀ and C-5ꢀ), 137.2 (C-1ꢀꢀ), 137.8 (C-5), 139.4 (C-16),
147.9 (C-17), 148.4 (C-1ꢀ), 156.8 (C-3). MS negative mode: 488 (20%
[M−H]−); positive mode: 490 (20% [M+H]+), 91 (100%, benzylic
cation).
2.12. Cylization of 3b
According to Section 2.10, hydrazone 3b (480 mg, 1.00 mmol)
and phenylselenyl bromide (236 mg, 1.00 mmol) were allowed to
react in acetonitrile (10 mL). After reduction, the crude product was
subjected to chromatographic purification on a silica gel column
with light petroleum/dichloromethane (50:50) as eluent, yielding
16 as a white solid (458 mg, 72%). mp 150–152 ◦C; Rf = 0.40 (CH2Cl2).
Anal. Calcd. for C39H44N2OSe: C, 73.68; H, 6.98. Found: C, 73.85; H,
7.21. 1H NMR (CDCl3): ı = 1.08 (s, 3H, 18-H3), 1.87 (d, 1H, J = 10.1 Hz)
and 2.86 (d, 1H, J = 10.1 Hz): 17a-H2, 2.23 (s, 3H, tolyl-CH3), 2.54 (m,
1H, 16-H), 2.81 (m, 2H, 6-H2), 3.12 (dd, 1H, J = 11.8 Hz, J = 7.2 Hz) and
3.39 (dd, 1H, J = 11.8 Hz, J = 2.8 Hz): 16a-H2, 3.90 (s, 1H, NH), 5.00
(s, 2H, OCH2), 6.69 (d, 1H, J = 2.6 Hz, 4-H), 6.75 (dd, 1H, J = 8.5 Hz,
J = 2.6 Hz, 2-H), 6.80 (d, 2H, J = 8.2 Hz, 2ꢀ-H and 6ꢀ-H), 6.98 (d, 2H,
J = 8.2 Hz, 3ꢀ-H and 5ꢀ-H), 7.14–7.21 (overlapping multiplets, 4H, 1-H
and three other aromatic protons), 7.28 (t, 1H, J = 7.2 Hz, 4ꢀꢀ-H), 7.35
(t, 2H, J = 7.2 Hz, 3ꢀꢀ-H and 5ꢀꢀ-H), 7.40 (d, 2H, J = 7.2 Hz, 2ꢀꢀ-H and 6ꢀꢀ-
H), 7.44 (m, 2H, aromatic protons) ppm. 13C NMR (CDCl3): ı = 17.6
(C-18), 20.5, 25.7, 26.2, 30.0, 31.1, 32.9, 35.4, 37.7, 38.5, 43.6, 47.6, 66.2
(C-16), 70.0 (OCH2), 70.3 (NCH2), 112.4 (C-2), 113.7 (2C, C-2ꢀ and C-
2.9. Reaction of 2 with 4-nitrophenylhydrazine hydrochloride
According to Section 2.1,
2
(375 mg, 1.00 mmol), 4-
nitrophenylhydrazine hydrochloride (190 mg, 1.00 mmol) and
sodium acetate (200 mg, 2.44 mmol) in methanol were allowed
to react. After work-up, 4d was obtained as a yellow solid
(510 mg, 90%). mp 132–134 ◦C; Rf = 0.73 (CH2Cl2). Anal. Calcd. for
C32H35N3O3: C, 75.41; H, 6.92. Found: C, 75.48; H, 7.06. 1H NMR
(CDCl3): ı = 1.28 (s, 3H, 18-H3), 2.84 (m, 2H, 6-H2), 4.97–5.04
(overlapping multiplets, 4H, OCH2 and 16a-H2), 5.80 (m, 1H, 16-H),
6.70 (d, 1H, J = 2.0 Hz, 4-H), 6.78 (dd, 1H, J = 8.6 Hz, J = 2.0 Hz, 2-H),
6.92 (d, 2H, J = 9.2 Hz, 2ꢀ-H and 6ꢀ-H), 7.19 (d, 1H, J = 8.6 Hz, 1-H),
7.29 (s, 1H, 17-H), 7.30 (t, 1H, J = 7.1 Hz, 4ꢀꢀ-H), 7.37 (t, 2H, J = 7.1 Hz,
3ꢀꢀ-H and 5ꢀꢀ-H), 7.41 (d, 2H, J = 7.1 Hz, 2ꢀꢀ-H and 6ꢀꢀ-H), 7.63 (s, 1H,
NH), 8.10 (d, 2H, J = 9.1 Hz, 3ꢀ-H and 5ꢀ-H) ppm. 13C NMR (CDCl3):
ı = 26.9, 27.2, 27.4, 30.3, 33.3, 39.3, 42.0, 42.3, 43.5, 50.9, 70.0
(OCH2), 111.2 (2C, C-2ꢀ and C-6ꢀ), 112.5 (C-2), 114.5 (C-4), 115.1
(C-16a), 126.2 (2C, C-3ꢀ and C-5ꢀ), 126.5 (C-4ꢀꢀ), 127.4 (2C, C-2ꢀꢀ and

E. Mernyák et al. / Steroids 74 (2009) 474–482
477
6ꢀ), 114.6 (C-4), 126.1 (2C), 126.2, 127.4 (2C), 127.8, 128.5 (2C), 128.7,
129.6 (2C, C-3ꢀ and C-5ꢀ), 131.2 (C-1ꢀꢀꢀ), 131.9 (2C), 132.9 (C-10), 137.2
(C-1ꢀꢀ), 137.8 (C-5), 145.8 (C-1ꢀ), 156.7 (C-3). MS positive mode: 635
(10% [M]+), 530 (60% [M−105]+), 91 (100%, benzylic cation).
J = 2.8 Hz): 16a-H2, 4.80 (s, 1H, NH), 5.03 (s, 2H, OCH2), 6.72 (d, 1H,
J = 2.4 Hz, 4-H), 6.79 (dd, 1H, J = 8.4 Hz, J = 2.4 Hz, 2-H), 6.84 (d, 2H,
J = 7.0 Hz, 2ꢀ-H and 6ꢀ-H), 7.17–7.23 (overlapping multiplets, 4H, 1-H
and three other aromatic protons), 7.31 (t, 1H, J = 7.3 Hz, 4ꢀꢀ-H), 7.37
(t, 2H, J = 7.3 Hz, 3ꢀꢀ-H and 5ꢀꢀ-H), 7.42 (overlapping multiplets, 4H,
2ꢀꢀ-H, 6ꢀꢀ-H and two other aromatic protons), 8.08 (d, 2H, J = 9.1 Hz,
3ꢀ-H and 5ꢀ-H) ppm. 13C NMR (CDCl3): ı = 17.6 (C-18), 25.6, 26.1,
29.9, 30.9, 32.5, 35.4, 37.6, 38.5, 43.6, 47.4, 66.4 (C-16), 70.0 (OCH2),
70.6 (NCH2), 111.4 (2C, C-2ꢀ and C-6ꢀ), 112.5 (C-2), 114.6 (C-4), 126.1,
126.2, 126.8, 127.4 (2C), 127.8 (2C), 128.5 (2C), 129.0 (2C), 131.2
(C-1ꢀꢀꢀ), 132.3 (2C), 132.7 (C-10), 137.3 (C-1ꢀꢀ), 137.8 (C-5), 139.5 (C-
4ꢀ), 153.4 (C-1ꢀ), 156.8 (C-3). MS positive mode: 666 (10% [M]+), 91
(100%, benzylic cation).
2.13. Cyclization of 3c
According to Section 2.10, hydrazone 3c (490 mg, 1.00 mmol)
and phenylselenyl bromide (236 mg, 1.00 mmol) were allowed to
react in acetonitrile (10 mL). After reduction, the crude product was
subjected to chromatographic purification on a silica gel column
with light petroleum/dichloromethane (50:50) as eluent, yielding
a 1:1.2 mixture of 18a (16␤) and 18b (16␣) (453 mg, 70%). In order
to obtain the pure diastereomers, the diastereoisomeric mixture of
18a and 18b was separated on a second silica gel column with the
same eluent.
2.15. Cyclization of 4a
Data for 18a: oil, Rf = 0.75 (CH2Cl2). Anal. Calcd. for
C39H41N3OSe: C, 72.43; H, 6.39. Found: C, 72.55; H, 6.42. 1H
NMR (CDCl3): ı = 1.12 (s, 3H, 18-H3), 2.05 and 2.78 (2×d, 2×1H,
J = 10.1 Hz): 17a-H2, 2.60 (m, 1H, 16-H), 2.83 (m, 2H, 6-H2), 3.10
(m, 1H) and 3.22 (dd, 1H, J = 11.9 Hz, J = 2.7 Hz): 16a-H2, 4.54 (s,
1H, NH), 5.03 (s, 2H, OCH2), 6.73 (d, 1H, J = 2.4 Hz, 4-H), 6.79 (dd,
1H, J = 8.6 Hz, J = 2.4 Hz), 6.88 (d, 2H, J = 8.1 Hz, 2ꢀ-H and 6ꢀ-H),
7.18–7.24 (overlapping multiplets, 4H, 1-H, aromatic protons), 7.32
(t, 1H, J = 7.1 Hz, 4ꢀꢀ-H), 7.39 (t, 2H, J = 7.1 Hz, 3ꢀꢀ-H and 5ꢀꢀ-H), 7.43
(overlapping multiplets, 6H, 3ꢀ-H, 5ꢀ-H, 2ꢀꢀ-H, 6ꢀꢀ-H and two other
aromatic protons) ppm. 13C NMR (CDCl3): ı = 17.6 (C-18), 25.7, 26.1,
29.9, 30.9, 32.5, 35.4, 37.6, 38.5, 43.6, 47.4, 66.3 (C-16), 70.0 (OCH2),
70.5 (NCH2), 100.8 (C-4ꢀ), 112.4 (2C, C-2ꢀ and C-6ꢀ), 112.5 (C-2),
114.6 (C-4), 120.2 (CN), 126.0, 126.1 (2C), 127.4 (2C), 127.8, 128.5
(2C), 129.0, 131.3 (C-1ꢀꢀꢀ), 132.2 (2C), 132.8 (C-10), 133.6 (2C, C-3ꢀ
and C-5ꢀ), 137.3 (C-1ꢀꢀ), 137.8 (C-5), 151.5 (C-1ꢀ), 156.8 (C-3). MS
negative mode: 117 (100% [NH-Ph-CN]−); positive mode: 646 (10%
[M−H]+), 530 (20% [M−117]+), 91 (100%, benzylic cation).
According to Section 2.10, hydrazone 4a (502 mg, 1.00 mmol)
and phenylselenyl bromide (236 mg, 1.00 mmol) were allowed to
react in acetonitrile (10 mL). After reduction, the crude product was
subjected to chromatographic purification on a silica gel column
with light petroleum/dichloromethane (50:50) as eluent, yielding
25 as an oil (442 mg, 71%). Rf = 0.37 (50% CH2Cl2/50% hexane). Anal.
Calcd. for C38H42N2OSe: C, 73.41; H, 6.81. Found: C, 73.22; H, 6.68.
1H NMR (CDCl3): ı = 1.29 (s, 3H, 18-H3), 2.26 and 2.54 (2×d,
2×1H, J = 12.5 Hz): 17a-H2, 2.82 (m, 2H, 6-H2), 2.94 (dd, 1H,
J = 12.0 Hz, J = 7.8 Hz) and 3.42 (dd, 1H, J = 12.0 Hz, J = 2.9 Hz): 16a-H2,
3.96 (s, 1H, NH), 5.04 (s, 2H, OCH2), 6.72 (d, 1H, J = 2.4 Hz, 4-H), 6.77
(overlapping multiplets, 2H, 2-H and 4ꢀ-H), 6.88 (d, 2H, J = 7.6 Hz,
2ꢀ-H and 6ꢀ-H), 7.16–7.22 (overlapping multiplets, 6H, 1-H, 3ꢀ-H, 5ꢀ-
H and three other aromatic protons), 7.32 (t, 1H, J = 7.3 Hz, 4ꢀꢀ-H),
7.38 (t, 2H, J = 7.3 Hz, 3ꢀꢀ-H and 5ꢀꢀ-H), 7.43 (d, 2H, J = 7.3 Hz, 2ꢀꢀ-H and
6ꢀꢀ-H), 7.47 (m, 2H) ppm. MS (EI); m/z (%): 622 (100, M+).
2.16. Cyclization of 4b
Data for 18b: oil, Rf = 0.73 (CH2Cl2). Anal. Calcd. for
C39H41N3OSe: C, 72.43; H, 6.39. Found: C, 72.58; H, 6.45. 1H
NMR (CDCl3): ı = 1.14 (s, 3H, 18-H3), 2.43 and 2.59 (2×d, 2×1H,
J = 10.4 Hz): 17a-H2, 2.83 (m, 2H, 6-H2), 3.12 (m, 1H) and 3.27 (dd,
1H, J = 12.0 Hz, J = 3.6 Hz): 16a-H2, 3.50 (m, 1H, 16-H), 5.03 (s, 2H,
OCH2), 5.43 (s, 1H, NH), 6.73 (d, 1H, J = 2.4 Hz, 4-H), 6.79 (dd, 1H,
J = 8.6 Hz, J = 2.4 Hz), 6.83 (d, 2H, J = 8.1 Hz, 2ꢀ-H and 6ꢀ-H), 7.18–7.24
(overlapping multiplets, 4H, 1-H and three other aromatic protons),
7.32 (t, 1H, J = 7.1 Hz, 4ꢀꢀ-H), 7.39 (t, 2H, J = 7.1 Hz, 3ꢀꢀ-H and 5ꢀꢀ-H),
7.43 (overlapping multiplets, 4H, 2ꢀꢀ-H, 6ꢀꢀ-H and two other aro-
matic protons), 7.49 (m, 2H, 3ꢀ-H and 5ꢀ-H) ppm. 13C NMR (CDCl3):
ı = 17.7 (C-18), 22.8, 25.6, 26.0, 27.1, 29.8, 35.5, 37.9, 38.1, 41.5, 43.7,
60.2 (C-16), 62.9 (NCH2), 70.0 (OCH2), 100.6 (C-4ꢀ), 112.4 (C-2),
112.6 (2C, C-2ꢀand C-6ꢀ), 114.5 (C-4), 120.2 (CN), 126.6 (2C), 127.2,
127.4 (2C), 127.8, 128.5 (2C), 129.2, 130.1 (C-1ꢀꢀꢀ), 132.6 (2C), 132.9
(C-10), 133.6 (2C, C-3ꢀ and C-5ꢀ), 137.3 (C-1ꢀꢀ), 137.9 (C-5), 151.2
(C-1ꢀ), 156.8 (C-3). MS negative mode: 117 (100% [NH-Ph-CN]−);
positive mode: 646 (10% [M−H]+), 530 (20% [M−117]+), 91 (100%,
benzylic cation).
According to Section 2.10, hydrazone 4b (480 mg, 1.00 mmol)
and phenylselenyl bromide (236 mg, 1.00 mmol) were allowed to
react in acetonitrile (10 mL). After reduction, the crude product was
subjected to chromatographic purification on a silica gel column
with light petroleum/dichloromethane (50:50) as eluent, yielding
26 as a white solid (503 mg, 79%). mp 85–87 ◦C; Rf = 0.41 (50%
CH2Cl2/50% hexane). Anal. Calcd. for C39H44N2OSe: C, 73.68; H,
6.98. Found: C, 73.95; H, 6.82. 1H NMR (CDCl3): ı = 1.28 (s, 3H, 18-
H3), 1.51 (d, J = 3.0 Hz) and 2.53 (d, J = 3.0 Hz): 17a-H2, 2.26 (s, 3H,
tolyl-CH3), 2.83 (m, 2H, 6-H2), 2.96 (dd, 1H, J = 11.9 Hz, J = 7.8 Hz) and
3.43 (dd, 1H, J = 11.9 Hz, J = 2.9 Hz): 16a-H2, 3.86 (s, 1H, NH), 5.05 (s,
2H, OCH2), 6.73 (d, 1H, J = 2.5 Hz, 4-H), 6.78 (overlapping multiplets,
3H, 2-H, 2ꢀ-H and 6ꢀ-H), 7.01 (d, 2H, J = 8.2 Hz, 3ꢀ-H and 5ꢀ-H), 7.17
(d, 1H, J = 8.6 Hz, 1-H), 7.20 (overlapping multiplets, 3H, 3ꢀꢀꢀ-H, 4ꢀꢀꢀ-H
and 5ꢀꢀꢀ-H), 7.33 (t, 1H, J = 7.1 Hz, 4ꢀꢀ-H), 7.39 (t, 2H, J = 7.1 Hz, 3ꢀꢀ-H
and 5ꢀꢀ-H), 7.43 (d, 2H, J = 7.1 Hz, 2ꢀꢀ-H and 6ꢀꢀ-H), 7.48 (overlapping
multiplets, 2H, 2ꢀꢀꢀ-H and 6ꢀꢀꢀ-H) ppm. 13C NMR (CDCl3): ı = 20.5,
26.6, 26.8, 28.6, 29.2, 30.2, 32.8, 35.0, 37.5, 38.4, 42.8, 44.9, 59.9 (C-
16), 61.1 (C-17a), 70.0 (OCH2), 112.4 (C-2), 113.5 (2C, C-2ꢀ and C-6ꢀ),
114.5 (C-4), 126.4 (2C), 127.4 (2C), 127.8, 128.4 (C-4ꢀ), 128.5 (2C),
128.9 (2C), 129.6 (2C, C-3ꢀ and C-5ꢀ), 131.7 (C-1ꢀꢀꢀ), 132.2 (2C), 133.0
(C-10), 137.3 (C-1ꢀꢀ), 138.1 (C-5), 146.0 (C-1ꢀ), 156.8 (C-3). MS posi-
tive mode: 635 (10% [M]+), 530 (50% [M−105]+), 91 (100%, benzylic
cation).
2.14. Cyclization of 3d
According to Section 2.10, hydrazone 3d (510 mg, 1.00 mmol)
and phenylselenyl bromide (236 mg, 1.00 mmol) were allowed to
react in acetonitrile (10 mL). After reduction the crude product
was subjected to chromatographic purification on a silica gel col-
umn with dichloromethane as eluent, yielding 17 as a yellow solid
(494 mg, 74%). mp 188–190 ◦C; Rf = 0.68 (CH2Cl2). Anal. Calcd. for
2.17. Cyclization of 4c
C
38H41N3O3Se: C, 68.46; H, 6.20. Found: C, 68.31; H, 6.42.
According to Section 2.10, hydrazone 4c (490 mg, 1.00 mmol)
and phenylselenyl bromide (236 mg, 1.00 mmol) were allowed to
react in acetonitrile (10 mL). After reduction, the crude product was
subjected to chromatographic purification on a silica gel column
1H NMR (CDCl3): ı = 1.15 (s, 3H, 18-H3), 2.09 and 2.77 (2×d,
2×1H, J = 10.4 Hz): 17a-H2, 2.63 (m, 1H, 16-H), 2.83 (m, 2H, 6-
H2), 3.08 (dd, 1H, J = 12.0 Hz, J = 6.9 Hz) and 3.19 (dd, 1H, J = 12.0 Hz,

478
E. Mernyák et al. / Steroids 74 (2009) 474–482
Scheme 1. Reagents and conditions: (a) Ph-NH-NH2·HCl or R-Ph-NH-NH2·HCl, NaOAc, MeOH, rt.
with light petroleum/dichloromethane (50:50) as eluent, yield-
ing 27 as an oil (472 mg, 73%). Rf = 0.78 (CH2Cl2). Anal. Calcd.
for C39H41N3OSe: C, 72.43; H, 6.39. Found: C, 72.58; H, 6.27. 1H
NMR (CDCl3): ı = 1.30 (s, 3H, 18-H3), 2.42 and 2.67 (2×d, 2×1H,
J = 10.7 Hz): 17a-H2, 2.83 (m, 2H, 6-H2), 2.90 (dd, 1H, J = 12.2 Hz,
J = 7.6 Hz) and 3.22 (dd, 1H, J = 12.2 Hz, J = 2.8 Hz): 16a-H2, 4.45 (s,
1H, NH), 5.04 (s, 2H, OCH2), 6.73 (d, 1H, J = 2.6 Hz, 4-H), 6.79 (dd,
1H, J = 8.5 Hz, J = 2.6 Hz, 2-H), 6.84 (d, 2H, J = 8.2 Hz, 2ꢀ-H and 6ꢀ-H),
7.17 (d, 1H, J = 8.5 Hz, 1-H), 7.22 (overlapping multiplets, 2H, aro-
matic protons), 7.32 (t, 1H, J = 7.1 Hz, 4ꢀꢀ-H), 7.38 (t, 2H, J = 7.1 Hz,
3ꢀꢀ-H and 5ꢀꢀ-H), 7.42–7.44 (overlapping multiplets, 5H, 2ꢀꢀ-H, 6ꢀꢀ-H
and three other aromatic protons) ppm. 13C NMR (CDCl3): ı = 26.6,
26.8, 28.5 (C-18), 29.0, 30.2, 32.4, 35.0, 37.5, 38.3, 42.8, 44.6, 60.0 (C-
16), 61.4 (NCH2), 70.0 (OCH2), 100.7 (C-4ꢀ), 112.5 (3C, C-2, C-2ꢀ and
C-6ꢀ), 114.5 (C-4), 120.2 (CN), 126.4, 126.8, 127.4 (2C), 127.8, 128.5
(2C), 129.0 (2C), 130.9 (C-1ꢀꢀꢀ), 132.5 (2C), 132.8 (C-10), 133.6 (2C,
C-3ꢀ and C-5ꢀ), 137.2 (C-1ꢀꢀ), 138.0 (C-5), 151.6 (C-1ꢀ), 156.8 (C-3).
MS negative mode: 530 (20% [M−117]−), 117 (100% [NH-PH-CN]−);
positive mode: 530 (20% [M−117]+), 91 (100%, benzylic cation).
3), 187.5 (C-17a). After reduction, the crude product was subjected
to chromatographic purification on a silica gel column with light
petroleum/dichloromethane (50:50) as eluent, yielding 28 as an oil
(467 mg, 70%). Rf = 0.65 (CH2Cl2). Anal. Calcd. for C38H41N3O3Se: C,
68.46; H, 6.20. Found: C, 68.72; H, 6.05. 1H NMR (CDCl3): ı = 1.31
(s, 3H, 18-H3), 2.41 and 2.71 (2×d, 2×1H, J = 10.6 Hz): 17a-H2, 2.65
(m, 1H, 16-H), 2.80 (m, 2H, 6-H2), 2.90 (dd, 1H, J = 12.2 Hz, J = 7.5 Hz)
and 3.19 (m, 1H): 16a-H2, 4.75 (s, 1H, NH), 5.04 (s, 2H, OCH2), 6.74
(d, 1H, J = 2.4 Hz, 4-H), 6.78 (overlapping multiplets, 3H, 2-H and
2ꢀ-H and 6ꢀ-H), 7.17 (d, 1H, J = 8.6 Hz, 1-H), 7.22 (overlapping multi-
plets, 3H, aromatic protons), 7.31 (t, 1H, J = 7.0 Hz, 4ꢀꢀ-H), 7.38 (t, 2H,
J = 7.0 Hz, 3ꢀꢀ-H and 5ꢀꢀ-H), 7.42–7.44 (overlapping multiplets, 4H,
2ꢀꢀ-H, 6ꢀꢀ-H and two other aromatic protons), 8.07 (d, 2H, J = 8.9 Hz,
3ꢀ-H and 5ꢀ-H) ppm. 13C NMR (CDCl3): ı = 26.6, 26.8, 28.5 (C-18),
29.0, 30.2, 32.3, 35.0, 37.4, 38.2, 42.7, 44.5, 60.1 (C-16), 61.4 (NCH2),
70.0 (OCH2), 111.2 (2C, C-2ꢀ, C-6ꢀ), 112.4 (C-2), 114.4 (C-4), 126.2
(2C), 126.5, 126.9, 127.4 (2C), 127.9, 128.5 (2C), 129.1 (2C), 130.7 (C-
1ꢀꢀꢀ), 132.6 (2C), 132.7 (C-10), 137.2 (C-1ꢀꢀ), 138.0 (C-5), 139.4 (C-4ꢀ),
153.4 (C-1ꢀ), 156.8 (C-3). MS positive mode: 666 (10% [M]+), 530
(50% [M−136]+), 91 (100%, benzylic cation).
2.18. Cyclization of 4d
3. Results and discussion
According to Section 2.10, hydrazone 4d (510 mg, 1.00 mmol)
and phenylselenyl bromide (236 mg, 1.00 mmol) were allowed to
react in acetonitrile (10 mL). The crude iminium salt 24d was fil-
tered off, washed with cold acetonitrile, and directly subjected
to NMR measurements. 1H NMR (CDCl3): ı = 1.68 (s, 3H, 18-H3),
2.85 (m, 2H, 6-H2), 3.36 (dd, 1H, J = 13.0 Hz, J = 9.1 Hz) and 3.54 (dd,
1H, J = 13.0 Hz, J = 2.6 Hz): 16a-H2, 4.38 (m, 1H, 16-H), 5.05 (s, 2H,
OCH2), 6.69 (d, 1H, J = 2.5 Hz, 4-H), 6.75 (dd, 1H, J = 8.7 Hz, J = 2.5 Hz,
2-H), 6.80 (d, 2H, J = 9.0 Hz, 2ꢀ-H and 6ꢀ-H), 7.07 (d, 1H, J = 8.7 Hz,
1-H), 7.13 (m, 2H, aromatic protons), 7.28 (t, 1H, J = 7.0 Hz, 4ꢀꢀ-H),
7.30–7.40 (overlapping multiplets, 6H, aromatic protons), 8.04 (d,
2H, J = 9.0 Hz, 3ꢀ-H and 5ꢀ-H), 9.04 (s, 1H, 17a-H), 11.7 (s, 1H, NH)
ppm. 13C NMR (CDCl3): ı = 26.5, 26.6, 28.4 (C-18), 28.7, 29.7, 29.9,
37.9, 40.9, 41.3, 41.4, 43.3, 61.5 (C-16), 70.0 (OCH2), 113.0 (C-2),
114.7 (C-4), 115.5 (2C, C-2ꢀ and C-6ꢀ), 126.0 (2C), 126.6, 127.4 (2C),
127.5 (C-1ꢀꢀꢀ), 127.9, 128.3, 128.5 (2C), 129.6 (2C), 130.1 (C-10), 133.4
(2C), 137.0 (C-1ꢀꢀ), 137.1 (C-5), 143.2 (C-4ꢀ), 146.9 (C-1ꢀ), 157.2 (C-
Steroidal hydrazones
3 and 4 were prepared from D-
secoaldehydes 1 and 2, which are available in four steps from
estrone or 13␣-estrone 3-benzyl ether, respectively [19]. In the
estrone series, reaction of the appropriate 4-substituted phenylhy-
drazine hydrochloride and sodium acetate in methanol took place
in a few minutes, and the products 3a–d were formed as precipi-
tates (Scheme 1). The resulting hydrazones 3a–d were washed with
methanol and subjected directly to NMR measurements, which
confirmed their structure. In the 13␣-estrone series, the reaction
with 4-tolylhydrazine hydrochloride proceeded in a different man-
ner. Under the same reaction conditions as described for the estrone
derivatives, no precipitate was formed. After stirring for 30 min,
the reaction mixture was cooled to −18 ◦C, when a light-yellow
precipitate of 4b appeared, which needed no purification. In both
estrone series, four differently substituted phenylhydrazones, 3a–d
and 4a–d, were synthesized.

E. Mernyák et al. / Steroids 74 (2009) 474–482
479
First, the estrone phenylhydrazones 3a–d were subjected to
cylization, with phenylselenyl bromide as electrophile trigger
(Scheme 3). Iminium salt formation was carried out in acetoni-
trile solution with 1 equiv. of phenylselenyl bromide, the reaction
mixture being stirred in an ice-water bath [9]. The intermedi-
ate seleniranium ion 10a–d was trapped via the imino N atom
of the phenylhydrazone 3a–d. Iminium salts 11–14 were formed
immediately. After the starting hydrazone 3a–d had reacted com-
pletely, the solvent was evaporated off. The crude salt 11–14 was
dissolved in dichloromethane, methanol was added and the C
N
double bond was reduced with 4 equiv. of potassium borohydride,
the reaction mixture being stirred for 24 h at room temperature,
which yielded 17-phenylamino- or substituted 17-phenylamino
aza-D-homoestrones 15–18. The selenocyclizations proceeded in a
stereoselective manner, furnishing the 16␤-phenylselenylmethyl-
aza-D-homo derivative 15–17, except for the cyclization of 3c,
which led to a diastereoisomeric mixture of the products 18a,b.
We recently reported the halogen- and phenylselenyl-induced
synthesis of nitrone dipoles from steroidal ␦-alkenyl oximes and
oxime ethers in both the estrone and the 13␣-estrone series [9].
Oximes or oxime ethers 19 of estrone 3-methyl ether behaved
as ambident nucleophiles: the attack of the N atom of the E iso-
mer led to the thermodynamically stable cyclic nitrone 21, but
the attack of the O atom of the Z isomer afforded the oxazepine
derivative 22. Additionally, a nonsymmetrical steroidal dimer 23
was formed by the intermolecular 1,3-dipolar cycloaddition of 21
and 22 (Scheme 4).
Scheme 2. Reagents and conditions: (a) PhSeBr, CH2Cl2, rt; (b) NaBH4, CH2Cl2,
MeOH, rt.
Tiecco et al. investigated the selenium-induced cyclizations
of ␥-alkenyl phenylhydrazones (5E, 5Z, 8, Scheme 2) [20]. The
hydride reductions of the initially formed iminium salts led to
pyrrolidinamine (6), piperidinamine (9) or tetrahydropyridazine
(7) derivatives. They concluded that the cyclization is a regiose-
lective process and that the structure of the product is strongly
determined by the geometry of the starting phenylhydrazone.
It was of particular interest to investigate whether the phenyl-
hydrazones of estrone benzyl ether give different types of products
under the conditions employed, since the internal nucleophile is an
ambident group: it can act with the imino or amino N as the reactive
site. Side-products containing a seven-membered D ring have not
Scheme 3. Reagents and conditions: (a) PhSeBr, CH3CN, ice-water bath, N2; (b) KBH4, CH2Cl2-MeOH, rt.

480
E. Mernyák et al. / Steroids 74 (2009) 474–482
Scheme 4. Reagents and conditions: (a) NBS or PhSeBr, MeCN, rt, N2.
been isolated. Nucleophilic attack of the amino N on C-16 would
lead to tetrahydrodiazepine derivatives, but attack of the imino N
on C-16a would lead to the azepanilamine derivative. To summarize
the results in the estrone series, no reaction involving the nucle-
ophilic attack of the amino N atom took place. Additionally, all the
reactions proceeded with similar rates and yields, irrespective of
the aromatic substituent.
Following the cyclizations in the estrone series, the reac-
tions of the 13␣-phenylhydrazones 4a–d were carried out under
the same reaction conditions as described for the estrone series
(Scheme 5). All the reactions were regio- and stereoselective,
16␣-phenylselenylmethyl aza-D-homo-13␣-estrones 25–28 being
formed in high yields. Similarly to the results obtained in the
estrone series, side-products with seven-membered D rings were
not isolated in the reactions of the 13␣-estrone phenylhydrazones.
The structures of the new products were determined with one-
and two-dimensional NMR and MALDI-MS techniques. The 1H NMR
spectra of 3a–d reveal the singlet of 17-H at 6.9 ppm and of NH at
>7 ppm. In the 13C NMR spectra, the signals of C-17 are observed in
the range 150–154 ppm. In the spectra of 4a–d, there is an upfield
shift of the singlet of 17-H (7.2 ppm) and a downfield shift of the
signal of C-17 (144–150 ppm). Fig. 1 shows representative 1H NMR
spectra of hydrazones 3c and 4c.
The 1H NMR spectra of the estrone aza-D-homo derivatives
15–17 and 18a display the two doublets of 17a-H2 at ∼1.9 and
2.8 ppm with a coupling constant of 10 Hz, the multiplet of 16-H
at ∼2.6 ppm, and 16a-H2 gives two readily distinguishable double
doublets at ∼3.1 and 3.3 ppm. In the 13C NMR spectra, C-16 and
C-17a are seen at ∼66.2 and 70.5 ppm. The 1H NMR spectra of the
13␣-estrone-D-homo derivatives 25–28 show similarities between
the pairs 25–26 and 27–28. The two doublets of 17a-H2 appear at
∼1.5 and 2.5 ppm, with different coupling constants in the spec-
tra of 25 and 26, but at 2.4 and 2.7 ppm in the spectra of 27 and
28. Fig. 2 shows representative 1H NMR spectra of aza-D homoe-
Scheme 5. Reagents and conditions: (a) PhSeBr, CH3CN, ice-water bath, N2; (b) KBH4, CH2Cl2-MeOH, rt.

E. Mernyák et al. / Steroids 74 (2009) 474–482
481
Fig. 1. 1H NMR spectra of hydrazones 3c and 4c.
strones 16 and 26. The downfield shift of C-16 (60 ppm) and C-17a
(61 ppm) of 13␣-estrone derivatives 25–28 can be observed from
a comparison of the 13C NMR spectra of the 13-epimer pairs. The
iminium salt 24d of the 4-nitrophenylhydrazone could be isolated
as a stable precipitate and subjected to NMR measurements. The
1H NMR spectrum of 24d displayed a multiplet at 4.38 ppm, due to
16-H, with the singlets of N-H and 17a-H at 9.04 and 11.73 ppm. In
the 13C spectrum, the signals of C-16 and C-17a appeared at 61.5 and
187.5 ppm. The configurations of C-16 were determined by means of
NOESY experiments. The results were in good agreement with our
previous findings on the halo- and selenocyclizations of oximes and
oxime ethers [8,9].
Neutral steroids are difficult to analyze by desorption/ionization
methods coupled with mass spectrometry, and there are only
a few literature reports on the analysis of derivatized steroids
through MALDI TOF mass spectrometry [21–23]. In our preliminary
study, heterocyclic estrone derivatives were efficiently measured
by MALDI TOF mass spectrometer using C70 fullerenes as matrix
without further chemical derivatization, since all the steroids con-
tained endo- or exocyclic amino N atoms which were capable of
protonation [9]. Accordingly, MALDI TOF measurements of the syn-
thesized phenylhydrazones 3 and 4 and their cyclic derivatives
16–18, and 26–28 were carried out in both negative and positive
mode with C70 fullerenes as matrix. In the negative mode detec-
tion, only fragment ions, but no molecular or quasimolecular ions
generally appeared, except for 3c,d and 4c,d, where the negatively
charged quasimolecular ions [M−H]− were detected. In some cases,
fragment ions which arose from the cleavage of the aminophenyl or
substituted aminophenyl function from the phenylhydrazone were
observed, e.g., at m/z = 117 Da [NHPhCN]− in the spectrum of 27. In
the positive mode detection, the positively charged quasimolecular
ions [M−H]+ formed through the cleavage of one hydrogen atom
from the amino group were detected in the spectra of phenylhy-
drazones 3a–d and 4a–d. Molecular ions appeared in many cases:
3c, 16, 17, 18a,b; 26 and 28. Additionally, two other fragment ions
were observed: that of the benzylic cation and one with a mass
of 530, from the steroid residue obtained by cleavage of the substi-
tuted phenylamino function, e.g., in the spectrum of 27. Fig. 3 shows
the representative mass spectrum of hydrazone 4d in negative
mode.
Fig. 2. 1H NMR spectra of aza D-homo derivatives 26 and 16.

482
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Secoaldehydes of estrone and 13␣-estrone were effec-
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cyclic iminium salts. Subsequent hydride reduction yielded new
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unprotected derivatives, which are promising candidates for
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TheauthorsthanktheHungarian ScientificResearchFund(OTKA
D 048294 and K 72309) for financial support.
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