Design, chemical synthesis and biological evaluation of 3-spiromorpholinone/3-spirocarbamate androsterone derivatives as inhibitors of 17β-hydroxysteroid dehydrogenase type 3

Article abstract of DOI:10.1016/j.bmc.2015.07.049

17β-Hydroxysteroid dehydrogenase type 3 (17β-HSD3) is a key enzyme involved in the biosynthesis of testosterone and dihydrotestosterone. These hormones are known to stimulate androgen-dependent prostate cancer. In order to generate effective inhibitors of androgen biosynthesis without androgenic effect, we synthesized a new family of 3-spiromorpholinone and 3-spirocarbamate androsterone derivatives bearing diversified hydrophobic groups. We also tested their inhibitory activity in a microsomal fraction of 17β-HSD3-containing rat testes, and their androgenic effect on androgen-sensitive LAPC-4 cells. From our first structure-activity relationship (SAR) study, we noted that compound 7e inhibited 17β-HSD3 (77% at 0.1 μM) compared to our reference compound RM-532-105 (76% at 0.1 μM), but exhibited a residual androgenic effect. A library of 7e analogue compounds was next synthesized in order to generate compounds with reduced androgenic activity. In this new SAR study, the sulfonamide compound 7e21 and the carboxamide compound 7e22 inhibited 17β-HSD3 (IC₅₀ = 28 and 88 nM, respectively). These two compounds were not androgenic and not cytotoxic even at the highest concentration tested, but their inhibitory activity decreased in intact LNCaP cells overexpressing 17β-HSD3 (LNCaP[17β-HSD3]). Structural modifications of these two lead compounds could however be tested to produce a second generation of 17β-HSD3 inhibitors.

Full text of DOI:10.1016/j.bmc.2015.07.049

Accepted Manuscript
Design, chemical synthesis and biological evaluation of 3-spiromorpholinone/
3-spirocarbamate androsterone deriva-tives as inhibitors of 17β-hydroxysteroid
dehydrogenase type 3
G.B. Djigoué, L.C. Kenmogne, J. Roy, R. Maltais, D. Poirier
PII:
S0968-0896(15)00633-1
DOI:
http://dx.doi.org/10.1016/j.bmc.2015.07.049
BMC 12478
Reference:
Bioorganic & Medicinal Chemistry
To appear in:
Received Date:
Revised Date:
Accepted Date:
20 May 2015
17 July 2015
25 July 2015
Please cite this article as: Djigoué, G.B., Kenmogne, L.C., Roy, J., Maltais, R., Poirier, D., Design, chemical
synthesis and biological evaluation of 3-spiromorpholinone/3-spirocarbamate androsterone deriva-tives as
inhibitors of 17β-hydroxysteroid dehydrogenase type 3, Bioorganic & Medicinal Chemistry (2015), doi: http://
dx.doi.org/10.1016/j.bmc.2015.07.049
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1
Design, chemical synthesis and biological evaluation of
3-spiromorpholinone/3-spirocarbamate androsterone deriva-
tives as inhibitors of 17β-hydroxysteroid dehydrogenase type 3
G. B. Djigoué; L. C. Kenmogne; J. Roy; R. Maltais; D. Poirier*
Laboratory of Medicinal Chemistry, CHU de Québec – Research Center (CHUL, T4)
and Laval UNiversity (Faculty of Medicine), Québec, Qc, Canada
* Address correspondence to this author to:
Laboratory of Medicinal Chemistry
CHU de Québec-Research Center (CHUL, T4-42)
2705 Laurier Boulevard, Québec, QC, G1V 4G2, Canada
Phone: (418) 654-2296
Fax: (418) 654-2298
E-mail: donald.poirier@crchul.ulaval.ca

2
Abstract
17β-Hydroxysteroid dehydrogenase type 3 (17β-HSD3) is a key enzyme involved in
the biosynthesis of testosterone and dihydrotestosterone. These hormones are known to
stimulate androgen-dependent prostate cancer. In order to generate effective inhibitors of
androgen biosynthesis without androgenic effect, we synthesized a new family of 3-
spiromorpholinone and 3-spirocarbamate androsterone derivatives bearing diversified
hydrophobic groups. We also tested their inhibitory activity in a microsomal fraction of 17β-
HSD3-containing rat testes, and their androgenic effect on androgen-sensitive LAPC-4 cells.
From our first structure-activity relationship (SAR) study, we noted that compound 7e
inhibited 17β-HSD3 (77 % at 0.1 µM) compared to our reference compound RM-532-105 (76
% at 0.1 µM), but exhibited a residual androgenic effect. A library of 7e analogues was next
synthesized in order to generate compounds with reduced androgenic activity. In this new
SAR study, the sulfonamide compound 7e21 and the carboxamide compound 7e22 inhibited
17β-HSD3 (IC50 = 28 and 88 nM, respectively). These two compounds were not androgenic
and not cytotoxic even at the highest concentration tested, but their inhibitory activity
decreased in intact LNCaP cells overexpressing 17β-HSD3 (LNCaP[17β-HSD3]). Structural
modifications of these two lead compounds could however be tested to produce a second
generation of 17β-HSD3 inhibitors.
Key words: Enzyme inhibitor, Androgen, Spiromorpholinone, Chemical synthesis, Prostate
cancer, 17β-Hydroxysteroid dehydrogenase
Abbreviations: ADT: androsterone; AR: androgen receptor; Ar: aryl group; DBD: DNA
binding domain; DCM: dichloromethane; DHT: dihydrotestosterone; DIPEA: N,N-
4
diisopropylethylamine; DMF : dimethylformamide; DMSO: dimethylsulfoxide; Δ -dione: 4-
androstene-3,17-dione; EtOAc: ethyl acetate; FBS: fetal bovine serum; 17β-HSD: 17β-
hydroxysteroid dehydrogenase; IR: infrared spectroscopy; KOt-Bu: potassium tert-butylate;
LDA: lithium diisopropylamide; LR-MS: low-resolution mass spectrometry; NADPH:
nicotinamide adenosine dinucleotide phosphate reduced form; NaHMDS: sodium
bis(trimethylsilyl)amide; NaOMe: sodium methoxide; NMO: N-methylmorpholine N-oxide;
NMR: nuclear magnetic resonance; Pd(dppf)Cl2: 1,1'-bis(diphenylphosphino)ferrocene-
palladium(II)dichloride; Ph: phenyl group; Rf: Retardation or retention factor; RT: room
temperature; SAR: structure-activity relationship; TLC: thin-layer chromatography; p-TSA:
para-toluene sulfonic acid; T: testosterone; TEA: triethylamine; THF: tetrahydrofurane;
TPAP: tetrapropylamonium perruthenate.

3
1. Introduction
Prostate cancer is the second leading cause of death in American men,1 and androgens
are well known to play an important role in the development, growth and progression of
prostate cancer.2-4 Among the enzymes involved in the biosynthesis of androgens in testes,5-8
4
17β-hydroxysteroid dehydrogenase type 3 (17β-HSD3) converts 4-androstene-3,17-dione (Δ -
dione) into the potent and major circulating androgen testosterone (T), in the presence of
cofactor NADPH.9 Both T and its metabolite dihydrotestosterone (DHT) bind the androgen
receptor (AR), which dimerizes, translocalizes in the nucleus and activates cell proliferation
(Fig. 1). Androgen deprivation has been recognized to be one of the most efficient treatments
for advanced stage prostate cancer.10 In fact, up to 80 % of patients with metastatic prostate
cancer respond to androgen deprivation treatments.11 Although many endocrine therapies are
now available to block either the formation of testicular T and the action of T and DHT on
AR,12-16 there are still some unmet medical needs for treatments using this kind of approach or
for new ones. In fact, several side effects such as hot flushes, erectile dysfunction, decreased
muscle mass, hypertension and hypokalemia are usually observed.17,18 As an example, a
recently approved cytochrome P450 17A1 (CYP17) inhibitor used for patients with metastatic
castration-resistant prostate cancer, abiraterone acetate, affects the formation of both
androgens and glucocorticoids, and replacement therapy is necessary when using this CYP17
inhibitor.19 According to these observations, and given that 17β-HSD3 acts in the last step of
the biosynthesis of T, we hope that the development of a potent inhibitor of 17β-HSD3
without an androgenic effect is a good therapeutic option for prostate cancer and other
androgen-dependent diseases.
Figure 1: Biosynthesis of testosterone (T) and dihydrotestosterone (DHT) from 4-androstene-
4
3,17-dione (Δ -dione) and their androgenic action. T and DHT bind to the androgen receptor
(AR) that dimerizes and translocalizes in the cell nucleus, it then binds to the DNA binding
domain (DBD) to co-activate cell proliferation.

4
From previous studies in our laboratory, androsterone (ADT) derivatives substituted at
position C-3 were identified as new inhibitors of 17β-HSD3, and many important criteria for
the inhibition of 17β-HSD3 were established.20-27 Although these inhibitors showed strong
blockage of the enzymatic activity, many of them produced an undesirable androgenic effect
on Shionogi (AR+) cells. In order to develop novel inhibitors of 17β-HSD3, we decided to
build a new ring system (cycle E) at position 3 of the ADT nucleus. In fact, introducing a 3-
spiroheterocyclic moiety is our strategy for adding rigidity and introducing diversified
hydrophobic groups with different orientations in space. This will enable the exploration of
the hydrophobic pocket identified from our studies as important for 17β-HSD3 inhibition.
Preliminary reports have shown the impact of the stereochemistry on the carbon adjacent to
the carbonyl of morpholinone.28,29 We herein present full details of the chemical synthesis and
characterization of new ADT derivatives substituted at position C-3 by a spiromorpholinone
4
or a spirocarbamate; the assessment of their ability to block the transformation of Δ -dione to
T by 17β-HSD3, and the evaluation of their androgenic effect on androgen-sensitive cells.
2. Material and methods
2.1. Chemical synthesis
Androsterone was purchased from Steraloids (Wilton, NH, USA). Benzyl bromide,
benzyl bromide derivatives, 4-(trifluoromethyl)benzenesulfonyl chloride, allyl bromide and
propargyl bromide were obtained from Sigma–Aldrich Canada Ltd. (Oakville, ON, Canada).
Solvents were obtained from Fisher Scientific (Montréal, QC, Canada). Reactions were run
under an inert (argon) atmosphere in oven-dried glassware. Analytical thin-layer
chromatography (TLC) was performed on 0.20-mm silica gel 60 F254 plates (Fisher
Scientific), and compounds were visualized using ammonium molybdate/sulfuric acid/water
(with heating). Flash column chromatography was performed with Silicycle R10030B 230–
400 mesh silica gel (Québec, QC, Canada). Infrared spectra (IR) were obtained from a thin
film of compound usually solubilized in DCM and deposited upon a NaCl pellet. They were
recorded with a Horizon MB 3000 ABB FTIR spectrometer (ABB, Canada) and only
characteristic bands are reported. Nuclear magnetic resonance (NMR) spectra were recorded
with a Bruker Avance 400 digital spectrometer (Billerica, MA, USA) and reported in ppm.
1
The residual H and 13C NMR signals of chloroform (7.26 and 77.00 ppm respectively) and
acetone (2.05 and 28.9 ppm, respectively) were used as internal references. Low-resolution
mass spectra (LR-MS) were recorded on a Shimadzu prominence apparatus (Kyoto, Japan)
equipped with a Shimadzu LCMS-2020. The name of compounds was generated using
ACD/Labs (chemist version) software (Toronto, ON, Canada).
2.1.1. General procedure for the synthesis of 3a-3d
In a Schlenk tube, the oxirane 226 (1.7 mmol) was dissolved in dry MeOH (15 mL) and
the freed amino acid methyl ester30 (17.2 mmol) was added. The Schlenk tube was screwed
down hermetically, stirred and heated at 90 °C for 21 h. The MeOH was then evaporated, the
crude reaction mixture dissolved in DCM and pre-adsorbed on silica gel, and then purified by
flash column chromatography (hexanes/EtOAc/TEA, 89:10:1).
2.1.1.1. Methyl N-{[(3R,5S,8R,9S,10S,13S,14S)-3-hydroxy-10,13-dimethylhexadecahydrospiro
[cyclopenta[a]phenanthrene-17,2'-[1,3]dioxolan]-3-yl]methyl}-L-leucinate (3a)
Amorphous solid (99 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 3479 and 3333
(OH and NH), 1736 (C=O) cm-1. 1H NMR (acetone-d6) δ 0.74 (m, 1H), 0.78 (s, CH3-19), 0.82

5
(s, 3H, CH3-18), 0.91 and 0.92 (2d, J = 6.9 Hz, (CH3)2- from iPr), 1.10-1.95 (unassigned CH
and CH2), 2.21 and 2.57 (2d of AB system, J = 11.6 Hz, CH2N), 3.23 (dd, J1 = 6.4 Hz, J2 = 8.2
Hz, CHC=O), 3.67 (s, OCH3), 3.83 (m, OCH2CH2O) ppm. 13C NMR (CDCl3) δ 11.2, 14.4,
20.3, 21.9, 22.6, 22.9, 24.8, 28.5, 30.7, 31.2, 31.7, 33.8, 34.2, 35.8, 36.0, 38.6, 40.6, 42.8,
46.0, 50.3, 51.7, 53.9, 59.3, 60.9, 64.5, 65.1, 69.9, 119.5, 176.3 ppm. LR-MS: calcd. for
C29H50NO5 [M+H]+ 492.36, found 492.35.
2.1.1.2. Methyl N-{[(3R,5S,8R,9S,10S,13S,14S)-3-hydroxy-10,13-dimethylhexadecahydrospiro
[cyclopenta[a]phenanthrene-17,2'-[1,3]dioxolan]-3-yl]methyl}-D-leucinate (3b)
Amorphous solid (99 % yield). Rf = 0.3 (hexanes/EtOAc, 1:1). IR (film) ν 3479 and 3333
(OH and NH), 1736 (C=O) cm-1. 1H NMR (CDCl3) δ 0.73 (s, CH3-19), 0.82 (m, 1H), 0.83 (s,
CH3-18), 0.90 and 0.93 (2d, J = 6.6 Hz, (CH3)2- from iPr), 0.96-2.00 (unassigned CH and
CH2), 2.17 and 2.62 (2d of AB system, J = 11.9 Hz, CH2N), 3.23 (tapp., J = 6.7 Hz, CHC=O),
3.72 (s, OCH3), 3.87 (m, OCH2CH2O) ppm. 13C NMR (CDCl3) δ 11.2, 14.4, 20.3, 21.9, 22.6,
22.8, 24.8, 28.4, 30.7, 31.2, 31.5, 33.6, 34.1, 35.8, 35.9, 38.6, 40.6, 42.7, 45.9, 50.3, 51.6,
53.8, 59.3, 60.8, 64.5, 65.1, 69.9, 119.4, 176.3 ppm. LR-MS: calcd. for C29H50NO5 [M+H]+
492.36, found 492.45.
2.1.1.3. Methyl N-{[(3R,5S,8R,9S,10S,13S,14S)-3-hydroxy-10,13-dimethylhexadecahydrospiro
[cyclopenta[a]phenanthrene-17,2'-[1,3]dioxolan]-3-yl]methyl}-L-phenylalaninate (3c)
Amorphous solid (77 % yield). Rf = 0.5 (hexanes/EtOAc, 1:1). IR (film) ν 3472 and 3340
(OH and NH), 1736 (C=O) cm-1. 1H NMR (acetone-d6) δ 0.72 (m, 1H), 0.75 (s, CH3-19), 0.82
(s, CH3-18), 0.95 (m, 1H), 1.12-1.97 (unassigned CH and CH2), 2.24 and 2.56 (2d of AB
system, J = 11.6 Hz, CH2N), 2.92 (m, CH2-Ph), 3.45 (t, J = 6.6 Hz, CHC=O), 3.62 (s, OCH3),
3.84 (m, OCH2CH2O), 7.25 (m, Ph) ppm. 13C NMR (CDCl3) δ 11.2, 14.4, 20.3, 22.7, 28.5,
30.7, 31.2, 31.5, 33.7, 34.2, 35.8, 35.9, 38.5, 39.8, 40.5, 46.0, 50.3, 51.7, 53.8, 59.3, 63.9,
64.5, 65.1, 70.0, 119.5, 126.7, 128.4 (2C), 129.1 (2C), 137.3, 175.0 ppm. LR-MS: calcd. for
C32H48NO5 [M+H]+ 526.35, found 526.50.
2.1.1.4. Methyl N-{[(3R,5S,8R,9S,10S,13S,14S)-3-hydroxy-10,13-dimethylhexadecahydrospiro
[cyclopenta[a]phenanthrene-17,2'-[1,3]dioxolan]-3-yl]methyl}-D-phenylalaninate (3d)
Amorphous solid (87 % yield). Rf = 0.43 (hexanes/EtOAc, 1:1). IR (film) ν 3472 and 3340
(OH and NH), 1736 (C=O) cm-1. 1H NMR (acetone-d6) δ 0.72 (m, 1H), 0.75 (s, CH3-19), 0.82
(s, CH3-18), 0.95 (m, 1H), 1.15-1.97 (unassigned CH and CH2), 2.24 and 2.56 (2d of AB
system, J = 11.6 Hz, CH2N), 2.92 (m, CH2-Ph), 3.43 (t, J = 6.6 Hz, CHC=O), 3.62 (s, OCH3),
3.84 (m, OCH2CH2O), 7.25 (m, Ph) ppm. 13C NMR (CDCl3) δ 11.2, 14.4, 20.4, 22.7, 28.4,
30.7, 31.2, 31.5, 33.6, 34.2, 35.8, 35.9, 38.4, 39.8, 40.5, 45.9, 50.3, 51.7, 53.8, 59.3, 64.0,
64.5, 65.1, 70.0, 119.4, 126.7, 128.4 (2C), 129.1 (2C), 137.3, 175.0 ppm. LR-MS: calcd. for
C32H48NO5 [M+H]+ 526.35, found 526.50.
2.1.2. Synthesis of methyl N-{[(3R,5S,8R,9S,10S,13S,14S)-3-hydroxy-10,13-dimethylhexa
decahydrospiro[cyclopenta[a]phenanthrene-17,2'-[1,3]dioxolan]-3-yl]methyl}glycinate
(3e)
To a mixture of glycine methyl ester hydrochloride (1.1 g, 8.8 mmol) and DIPEA (2.2
g, 17.5 mmol) was added anhydrous MeOH (20 mL) and the solution was stirred for 30 min
in a Schlenk tube at room temperature. Oxirane 226 (0.3 g, 0.9 mmol) was then added and the
solution heated to 95 °C for 22 h. The solution was cooled to room temperature, filtered on a
Büchner and the filtrate was evaporated. The crude mixture (1.8 g) was dissolved in DCM,
adsorbed on silica gel (7 g) (dry-pack) and then eluted with a mixture of hexanes/EtOAc/TEA

6
(70:30:1) to give 3e. Amorphous solid (53 % yield). Rf = 0.17 (hexanes/EtOAc, 3:7); IR
1
(film) ν 3464 and 3348 (OH and NH), 1744 (C=O) cm-1. H NMR (CDCl3) δ 0.74 (s, CH3-
19), 0.83 (s, CH3-18), 0.85-1.97 (unassigned CH and CH2), 2.50 (s, CH2N), 3.45 (s,
13
NCH2C=O), 3.73 (s, OCH3), 3.83-3.95 (m, OCH2CH2O) ppm. C NMR (acetone-d6) δ 10.8,
13.9, 20.3, 22.4, 30.7, 31.4, 31.5, 33.3, 33.7, 33.9, 35.8, 35.9, 38.5, 40.5, 45.8, 50.3, 50.7,
51.2, 54.4, 61.1, 64.2, 64.8, 70.0, 118.9, 172.7 ppm. LR-MS: calcd. for C25H41NO5 [M+H]+
436.30, found 436.35.
2.1.3. Synthesis of the spiromorpholinones 4a-4e
To a solution of MeONa (0.37 mmol) in anhydrous THF (24 mL) at 0 °C was added a
solution of the amino alcohol (3a-3e) (0.61 mmol) in dry THF (28 mL) and under an argon
atmosphere. The solution was stirred for 2 h at room temperature and the reaction mixture
was quenched with a saturated ammonium chloride aqueous solution. The crude product was
extracted four times with EtOAc and the organic layer was dried with anhydrous MgSO4,
filtered over vacuum and dried under reduced pressure. The residue was adsorbed on silica gel
and purified by flash column chromatography with a mixture of hexanes/EtOAc/TEA
(80:20:1).
2.1.3.1. (3'R,5'S,5''S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-5''-(2-methylpropyl)
tetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (4a)
Amorphous solid (89 % yield). Rf = 0.21 (hexanes/EtOAc, 8:2). IR (film) ν 3340 (NH) and
1
1720 (C=O) cm-1. H NMR (acetone-d6) δ 0.79 (m, 1H), 0.83 (s, CH3-19 and CH3-18), 0.89
and 0.92 (2d, J = 6.6 Hz, (CH3)2- from iPr), 0.95-1.98 (unassigned CH and CH2), 2.83 and
2.93 (2d of AB system, J = 13.5 Hz, CH2N), 3.43 (dd, J1 = 4.0 Hz, J2 = 9.6 Hz, NCHC=O),
3.85 (m, OCH2CH2O) ppm. 13C NMR (CDCl3) δ 11.3, 14.4, 20.3, 21.0, 22.6, 23.4, 24.4, 28.1,
30.6, 31.5, 32.9, 34.1, 35.7, 36.0, 38.2, 39.2, 39.6, 41.4, 45.9, 50.1, 52.5, 53.3, 55.5, 64.5,
65.1, 82.3, 119.4, 171.9 ppm. LR-MS: calcd. for C28H46NO4 [M+H]+ 460.33, found 460.45.
2.1.3.2.
(3'R,5'S,5''R,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-5''-(2-methylpropyl)
tetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (4b)
Amorphous solid (84 % yield). Rf = 0.23 (hexanes/EtOAc, 8:2). IR (film) ν 3340 (NH) and
1
1720 (C=O) cm-1. H NMR (CDCl3) δ 0.76 (s, CH3-19), 0.83 (s, CH3-18), 0.92 and 0.96 (2d,
J = 6.3 Hz, (CH3)2- from iPr), 0.80-1.99 (unassigned CH and CH2), 2.80 and 2.88 (2d of AB
system, J = 13.5 Hz, CH2N), 3.49 (dd, J1 = 3.5 Hz, J2 = 10.0 Hz, NCHC=O), 3.89 (m,
OCH2CH2O) ppm. 13C NMR (CDCl3) δ 11.3, 14.4, 20.3, 20.9, 22.7, 23.4, 24.5, 28.2, 30.6,
31.5, 32.6, 34.2, 35.7, 36.0, 38.3, 39.3, 39.7, 41.5, 45.9, 50.1, 52.6, 53.4, 55.5, 64.5, 65.1,
82.3, 119.4, 171.9 ppm. LR-MS: calcd. for C28H46NO4 [M+H]+ 460.33, found 460.45.
2.1.3.3. (3'R,5'S,5''S,8'R,9'S,10'S,13'S,14'S)-5''-benzyl-10',13'-dimethyltetradecahydro-
2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-[1,4]oxazinan]-
6''-one (4c)
Amorphous solid (62 % yield). Rf = 0.15 (hexanes/EtOAc, 8:2). IR (film) ν 3340 (NH) and
1
1720 (C=O) cm-1. H NMR (acetone-d6) δ 0.75 (m, 1H), 0.78 (s, CH3-19), 0.82 (s, CH3-18),
0.90-1.99 (unassigned CH and CH2), 2.93 (m, CH2N), 3.05 and 3.17 (2m, 2H, CH2Ph), 3.72
(dd, J1 = 3.9 Hz, J2 = 8.1 Hz, NCHC=O), 3.84 (m, OCH2CH2O), 7.22 (m, 1H of Ph), 7.29 (m,
4H of Ph) ppm. 13C NMR (CDCl3) δ 11.3, 14.4, 20.3, 22.6, 28.0, 30.6, 30.9, 31.1, 32.9, 34.2,

7
35.7, 35.9, 38.0 (2C), 39.2, 45.9, 50.1, 52.7, 53.3, 58.7, 64.5, 65.1, 82.8, 119.4, 127.0, 128.7
(2C), 129.5 (2C), 137.2, 170.8 ppm. LR-MS: calcd. for C31H44NO4 [M+H]+ 494.32, found
494.45.
2.1.3.4. (3'R,5'S,5''R,8'R,9'S,10'S,13'S,14'S)-5''-benzyl-10',13'-dimethyltetradecahydro-
2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-[1,4]oxazinan]-
6''-one (4d)
Amorphous solid (65 % yield). Rf = 0.09 (hexanes/EtOAc 8:2). IR (film) ν 3340 (NH) and
1720 (C=O) cm-1. 1H NMR (acetone-d6) δ 0.77 (s, CH3-19), 0.82 (s, CH3-18), 0.85 (m, 1H),
0.88-1.98 (unassigned CH and CH2), 2.85 (m, CH2N), 3.05 and 3.15 (2m, CH2Ph), 3.74 (dd,
J1= 4.1 Hz, J2 = 7.6 Hz, NCHC=O), 3.84 (m, OCH2CH2O), 7.24 (m, 1H of Ph), 7.27 (m, 4H
of Ph) ppm. 13C NMR (CDCl3) δ 11.3, 14.4, 20.3, 22.7, 28.0, 30.6, 31.0, 31.3, 32.5, 34.2,
35.7, 35.9, 37.7, 37.9, 39.6, 45.9, 50.1, 52.7, 53.3, 58.7, 64.5, 65.1, 82.7, 119.4, 127.1, 128.8
(2C), 1.6 (2C), 137.2, 170.8 ppm. LR-MS: calcd. for C31H44NO4 [M+H]+ 494.32, found
494.50.
2.1.3.5. (3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyltetradecahydro-2'H,6''H-dispiro
[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-[1,4]oxazinan]-6''-one (4e)
Amorphous solid (52 % yield). Rf = 0.16 (hexanes/EtOAc, 3:7). IR (film) ν 3340 (NH) and
1
1728 (C=O) cm-1. H NMR (CDCl3) δ 0.78 (s, CH3-19), 0.83 (s, CH3-18), 0.84-1.99
(unassigned CH and CH2), 2.82 (s, CH2N), 3.63 (s, NCH2C=O), 3.90 (m, OCH2-CH2O) ppm.
13C NMR (CDCl3) δ 11.3, 14.4, 20.4, 22.7, 28.1, 30.6, 31.0, 31.3, 32.7, 34.2, 35.7, 36.0, 38.0,
39.4, 45.9, 47.6, 50.1, 53.1, 53.4, 64.6, 65.2, 82.5, 119.4, 168.8 ppm. LR-MS: calcd. for
C24H38NO4 [M+H]+ 404.27, found 404.35.
2.1.4. Synthesis of N-substituted spiromorpholinones 6a-6e and 6e1-6e22
To a solution of 4a-4e (0.1 mmol) in dry DCM (5 mL) was added drop-wise DIPEA
(0.17 mmol) in a Schlenk tube. The tube was screwed down, stirred and heated at 75 °C for
10 min, and the reaction mixture was cooled to room temperature and benzyl bromide (1.7
mmol) was added to the solution. The reaction mixture was stirred and heated at 75 °C for 48
h. After cooling the mixture, silica gel was added to the crude mixture, the solvent was
evaporated under reduced pressure and the residue was purified by flash chromatography with
a mixture of hexanes/EtOAc/TEA (95:5:1).
2.1.4.1.
(3'R,5'S,5''S,8'R,9'S,10'S,13'S,14'S)-4''-benzyl-10',13'-dimethyl-5''-(2-methyl
propyl)tetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-
3',2''-[1,4]oxazinan]-6''-one (6a)
Amorphous solid (63 % yield). Rf = 0.83 (hexanes/EtOAc, 1:1). 1H NMR (acetone-d6) δ 0.74
(s, CH3-19), 0.81 (s, CH3-18), 0.93 and 0.98 (2d, J = 6.7 Hz, (CH3)2- from iPr), 1.00-2.00
(unassigned CH and CH2), 2.27 and 2.74 (2d of AB system, J = 12.5 Hz, CH2N), 3.17 (dd, J1
= 2.7 Hz, J2 = 7.0 Hz, NCHC=O), 3.23 and 4.07 (2d of AB system, J = 13.7 Hz, NCH2-Ph),
3.83 (m, OCH2CH2O), 7.28 (m, 1H of Ph), 7.37 (m, 4H of Ph) ppm.
2.1.4.2.
(3'R,5'S,5''R,8'R,9'S,10'S,13'S,14'S)-4''-benzyl-10',13'-dimethyl-5''-(2-methyl
propyl)tetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-
3',2''-[1,4]oxazinan]-6''-one (6b)

8
1
Amorphous solid (49 % yield). Rf = 0.83 (hexanes/EtOAc, 1:1). H NMR (CDCl3) δ 0.68 (s,
CH3-19), 0.82 (s, CH3-18), 0.95 and 0.99 (2d, J = 6.7 Hz, (CH3)2- from iPr), 0.80-2.11
(unassigned CH and CH2), 2.16 and 2.64 (2d of AB system, J = 12.4 Hz, CH2N), 3.12 and
4.02 (2d of AB system, J = 13.5 Hz, NCH2-Ph), 3.18 (dd, J1 = 2.8 Hz, J2 = 7.0 Hz,
NCHC=O), 3.89 (m, OCH2CH2O), 7.33 (m, Ph) ppm. 13C NMR (CDCl3) δ 11.4, 14.4, 20.3,
22.1, 22.7, 23.9, 25.0, 28.3, 30.7, 31.1, 32.0, 32.7, 34.2, 35.7, 36.0, 38.3, 38.6, 39.8, 45.9,
50.1, 53.4, 57.8, 58.2, 63.0, 64.6, 65.1, 81.3, 119.4, 127.4, 128.5 (4C), 137.8, 171.4 ppm.
2.1.4.3. (3'R,5'S,5''S,8'R,9'S,10'S,13'S,14'S)-4'',5''-dibenzyl-10',13'-dimethyltetra deca
hydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6c)
Amorphous solid (70 % yield). Rf = 0.75 (hexanes/EtOAc, 1:1). 1H NMR (acetone-d6) δ 0.62
(s, CH3-19), 0.68 (m, 1H), 0.79 (s, CH3-18), 0.88-1.95 (unassigned CH and CH2), 2.21 and
2.58 (2d of AB system, J = 12.5 Hz, CH2N), 3.28 and 4.39 (2d of AB system, J = 13.8 Hz,
NCH2-Ph), 3.36 (m, CH2Ph), 3.51 (dd, J1 = 3.0 Hz, J2 = 5.1 Hz, NCHC=O), 3.85 (m,
OCH2CH2O), 7.33 (m, Ph) ppm. 13C NMR (acetone-d6) δ 10.7, 13.9, 20.2, 22.4, 27.9, 29.5,
30.6, 31.2, 33.1, 33.9, 35.2, 35.7, 37.9, 39.5, 45.7, 50.2, 54.0, 57.5, 57.7, 61.0, 64.2, 64.8,
65.9, 80.5, 118.8, 126.4, 127.1, 127.8 (2C), 128.3 (2C), 128.5 (2C), 130.4 (2C), 138.0, 138.1,
169.5 ppm.
2.1.4.4. (3'R,5'S,5''R,8'R,9'S,10'S,13'S,14'S)-4'',5''-dibenzyl-10',13'-dimethyltetra deca
hydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6d)
Amorphous solid (61 % yield). Rf = 0.75 (hexanes/EtOAc, 1:1). 1H NMR (acetone-d6) δ 0.62
(s, CH3-19), 0.68 (m, 1H), 0.78 (s, CH3-18), 0.80-1.97 (unassigned CH and CH2), 2.21 and
2.59 (2d of AB system, J = 12.4 Hz, CH2N), 3.28 and 4.41 (2d of AB system, J = 13.8 Hz,
NCH2-Ph), 3.38 (m, CH2Ph), 3.50 (dd, J1 = 3.0 Hz, J2 = 5.1 Hz, NCHC=O), 3.83 (m,
OCH2CH2O), 7.30 (m, Ph) ppm. 13CNMR (acetone-d6) δ 10.8, 13.9, 20.2, 22.4, 27.6, 29.5,
30.6, 31.1, 31.2, 32.7, 33.9, 35.0, 35.7, 37.3, 39.9, 45.7, 50.2, 54.1, 57.5, 57.8, 64.2, 64.8,
65.8, 80.5, 118.8, 126.5, 127.1, 127.8 (2C), 128.4 (2C), 128.6 (2C), 130.5 (2C), 137.9, 138.1,
169.4 ppm.
2.1.4.5.
(3'R,5'S,8'R,9'S,10'S,13'S,14'S)-4''-benzyl-10',13'-dimethyltetradecahydro-
2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-[1,4]oxazinan]-
6''-one (6e)
1
Amorphous solid (70 % yield). Rf = 0.83 (hexanes/EtOAc, 1:1). H NMR (CDCl3) δ 0.72 (s,
CH3-19), 0.82 (s, CH3-18), 0.80-1.99 (unassigned CH and CH2), 2.41 (s-broad, CH2N), 3.26
(s, NCH2C=O), 3.50 and 3.54 (2d of AB system, J = 13.3 Hz, NCH2Ph), 3.89 (m,
OCH2CH2O), 7.31 (m, Ph) ppm. 13C NMR (CDCl3) δ 11.4, 14.4, 20.3, 22.7, 28.1, 30.7, 31.0,
31.8, 32.9, 34.2, 35.7, 36.0, 38.4, 39.6, 45.9, 50.1, 53.4, 55.4, 59.8, 61.4, 64.6, 65.1, 82.5,
119.4, 127.6, 128.5 (2C), 128.7 (2C), 136.7, 168.3 ppm.
2.1.4.6. (3'R,5'S,8'R,9'S,10'S,13'S,14'S)-4''-(4-bromobenzyl)-10',13'-dimethyltetradeca
hydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6e1)
Amorphous solid (87 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1724 (C=O) cm-1.
1H NMR (CDCl3) δ 0.73 (s, CH3-19), 0.83 (s, CH3-18), 0.80-2.00 (unassigned CH and CH2),

9
2.39 (s, CH2N), 3.26 (s, NCH2C=O), 3.46 (s, CH2-Ph), 3.89 (m, OCH2CH2O), 7.19 (d, J = 8.3
Hz, 2H of Ph), 7.47 (d, J = 8.3 Hz, 2H of Ph) ppm. 13C NMR (CDCl3) δ 11.4, 14.4, 20.3, 22.7,
28.1, 30.6, 31.0, 31.8, 32.9, 34.2, 35.7, 36.0, 38.4, 39.6, 45.9, 50.1, 53.4, 55.4, 59.7, 60.6,
64.5, 65.1, 82.4, 119.4, 121.5, 130.3 (2C), 131.7 (2C), 135.8, 167.9 ppm. LR-MS: calcd. for
C31H4379BrNO4 [M+H]+ 572.23, found 572.35.
2.1.4.7.
(3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-[4-(trifluoromethoxy)
benzyl]tetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-
3',2''-[1,4]oxazinan]-6''-one (6e2)
Amorphous solid (95 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 1732 (C=O) cm-1.
1H NMR (CDCl3) δ 0.74 (s, CH3-19), 0.83 (s, CH3-18), 0.80-1.99 (unassigned CH and CH2),
2.43 (s, CH2N), 3.25 (s, NCH2C=O), 3.51 (d, J = 2.4 Hz, CH2-Ph), 3.89 (m, OCH2CH2O),
7.19 (d, J = 8.0 Hz, 2H of Ph), 7.34 (d, J = 8.6 Hz, 2H of Ph) ppm. 13C NMR (CDCl3) δ 11.4,
14.4, 20.3, 22.6, 28.1, 30.6, 31.0, 31.8, 32.9, 34.2, 35.7, 36.0, 38.4, 39.6, 45.9, 50.1, 53.4,
55.3, 59.8, 60.5, 64.5, 65.1, 82.4, 119.4, 120.4 (d, JC-F = 257.1 Hz), 121.0 (2C), 129.9 (2C),
135.5, 148.6, 167.9 ppm. LR-MS: calcd. for C32H43F3NO5 [M+H]+ 578.30, found 578.40.
2.1.4.8. (3'R,5'S,8'R,9'S,10'S,13'S,14'S)-4''-(biphenyl-4-ylmethyl)-10',13'-dimethyltetra
decahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6e3)
Amorphous solid (91 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 1728 (C=O) cm-1.
1H NMR (CDCl3) δ 0.73 (s, CH3-19), 0.83 (s, CH3-18), 0.82-1.99 (unassigned CH and CH2),
2.46 (s, CH2N), 3.30 (s, NCH2C=O), 3.56 (d, J = 2.2 Hz, CH2-Ph), 3.89 (m, OCH2CH2O),
13
7.36 (m, 3H of Ar), 7.45 (m, 2H of Ar), 7.59 (m, 4H of Ar) ppm. C NMR (CDCl3) δ 11.4,
14.4, 20.4, 22.7, 28.1, 30.7, 31.0, 31.9, 32.9, 34.2, 35.8, 36.1, 38.5, 39.6, 46.0, 50.2, 53.4,
55.5, 59.8, 61.0, 64.6, 65.1, 82.5, 119.4, 127.0 (2C), 127.3 (2C), 127.4, 128.8 (2C), 129.1
(2C), 135.7, 140.5, 140.7, 168.2 ppm. LR-MS: calcd. for C37H48NO4 [M+H]+ 570.35, found
570.45.
2.1.4.9.
(3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-(2-phenylethyl)tetradeca
hydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6e4)
Amorphous solid (41 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1732 (C=O) cm-1.
1H NMR (CDCl3) δ 0.77 (s, CH3-19), 0.83 (s, CH3-18), 0.75-2.05 (unassigned CH and CH2),
2.50 (s, CH2N), 2.61 (tapp., J = 7.5 Hz, CH2-Ph), 2.78 (tapp., J = 7.6 Hz, NCH2CH2Ph), 3.29 (s,
NCH2C=O), 3.89 (m, OCH2CH2O), 7.20 (m, 3H of Ph), 7.28 (m, 2H of Ph) ppm. 13C NMR
(CDCl3) δ 11.4, 14.4, 20.4, 22.7, 28.1, 30.6, 31.0, 31.8, 32.9, 33.2, 34.2, 35.8, 36.0, 38.5,
39.6, 45.9, 50.2, 53.4, 55.3, 58.6, 60.4, 64.6, 65.1, 82.3, 119.4, 126.3, 128.4 (2C), 128.6 (2C),
139.5, 168.3 ppm. LR-MS: calcd. for C32H46NO4 [M+H]+ 508.33, found 508.45.
2.1.4.10. (3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-(3-phenylpropyl)tetradeca
hydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6e5)
Amorphous solid (73 % yield). Rf = 0.9 (hexanes/EtOAc, 1:1). IR (film) ν 1732 (C=O) cm-1.
1H NMR (CDCl3) δ 0.78 (s, CH3-19), 0.83 (s, CH3-18), 0.80-2.00 (unassigned CH and CH2),
2.34 (t, J = 7.0 Hz, NCH2CH2CH2Ph), 2.43 (s, CH2N), 2.66 (t, J = 7.5 Hz, NCH2CH2CH2Ph),
3.21 (s, NCH2C=O), 3.89 (m, OCH2CH2O), 7.18 (m, 3H of Ph), 7.28 (m, 2H of Ph) ppm. 13C
NMR (CDCl3) δ 11.4, 14.4, 20.4, 22.7, 28.1, 28.2, 30.7, 31.0, 31.9, 32.9, 33.0, 34.2, 35.8,

10
36.1, 38.6, 39.6, 46.0, 50.2, 53.4, 55.5, 56.0, 60.3, 64.6, 65.1, 82.3, 119.4, 125.9, 128.4 (4C),
141.7, 168.4 ppm. LR-MS: calcd. for C33H48NO4 [M+H]+ 522.35, found 522.45.
2.1.4.11.
(3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-{2-[4-(trifluoromethyl)
phenyl]ethyl}tetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]
phenanthrene-3',2''-[1,4]oxazinan]-6''-one (6e6)
Amorphous solid (53 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1732 (C=O) cm-1.
1H NMR (CDCl3) δ 0.79 (s, CH3-19), 0.83 (s, CH3-18), 0.80-2.00 (unassigned CH and CH2),
2.49 (s, CH2N), 2.63 (t, J = 7.3 Hz, CH2-Ph), 2.83 (t, J = 7.3 Hz, NCH2CH2Ph), 3.28 (d, J =
1.5 Hz, NCH2C=O), 3.89 (m, OCH2CH2O), 7.30 (d, J = 8.0 Hz, 2H of Ph), 7.54 (d, J = 8.1
Hz, 2H of Ph) ppm. 13C NMR (CDCl3) δ 11.4, 14.4, 20.3, 22.6, 28.1, 30.6, 30.6, 31.0, 31.8,
32.9, 33.0, 34.2, 35.7, 36.0, 38.5, 39.6, 45.9, 50.1, 53.4, 55.2, 60.4, 64.5, 65.1, 82.3, 119.4,
124.2 (q, JC-F = 272.0 Hz), 125.3 (q, JC-C-C-F = 3.6 Hz) (2C), 128.7 (JC-C-F = 32.5 Hz), 129.0
(2C), 143.6, 168.0 ppm. LR-MS: calcd. for C33H45F3NO4 [M+H]+ 576.32, found 576.40.
2.1.4.12. (3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-[4-(trifluoromethyl)benzyl]
tetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6e7)
Amorphous solid (85 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1732 (C=O) cm-1.
1H NMR (CDCl3) δ 0.73 (s, CH3-19), 0.83 (s, CH3-18), 0.82-2.00 (unassigned CH and CH2),
2.42 (s, CH2N), 3.28 (s, NCH2C=O), 3.57 (d, J = 2.7 Hz, CH2-Ph), 3.89 (m, OCH2CH2O),
7.44 (d, J = 8.0 Hz, 2H of Ph), 7.61 (d, J = 8.1 Hz, 2H of Ph) ppm. 13C NMR (CDCl3) δ 11.4,
14.4, 20.3, 22.6, 28.1, 30.6, 31.0, 31.9, 32.9, 34.2, 35.7, 36.0, 38.4, 39.6, 45.9, 50.1, 53.4,
55.4, 59.8, 60.8, 64.5, 65.1, 82.4, 119.4, 124.1 (q, JC-F = 272.0 Hz), 125.5 (JC-C-C-F = 3.8 Hz)
(2C), 128.8 (2C), 130.0 (JC-C-F = 32.4 Hz), 140.9, 167.8 ppm. LR-MS: calcd. for C32H43F3NO4
[M+H]+ 562.31, found 562.40.
2.1.4.13. (3'R,5'S,8'R,9'S,10'S,13'S,14'S)-4''-[4-(benzyloxy)benzyl]-10',13'-dimethyltetra
decahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6e8)
Amorphous solid (70 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 1732 (C=O) cm-1.
1H NMR (CDCl3) δ 0.73 (s, CH3-19), 0.83 (s, CH3-18), 0.82-2.00 (unassigned CH and CH2),
2.40 (s, CH2N), 3.24 (s, NCH2C=O), 3.45 (d, J = 2.6 Hz, CH2-Ph), 3.89 (m, OCH2CH2O),
5.07 (s, CH2O), 6.95 (d, J = 8.6 Hz, 2H of Ph), 7.21 (d, J = 8.6 Hz, 2H of Ph), 7.39 (m, 5H of
13
Ph) ppm. C NMR (CDCl3) δ 11.4, 14.4, 20.3, 22.7, 28.1, 30.7, 31.0, 31.9, 32.9, 34.2, 35.8,
36.0, 38.4, 39.6, 46.0, 50.1, 53.4, 55.4, 59.7, 60.7, 64.6, 65.1, 70.0, 82.5, 114.8 (2C), 119.4,
127.5 (2C), 128.0, 128.6 (2C), 128.9, 129.9 (2C), 136.9, 158.3, 168.3 ppm. LR-MS: calcd. for
C38H50NO5 [M+H]+ 600.36, found 600.45.
2.1.4.14. (3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-(4-phenylbutyl)tetradeca
hydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6e9)
Amorphous solid (69 % yield). Rf = 0.8 (hexanes/EtOAc, 1:1). IR (film) ν 1732 (C=O) cm-1.
1H NMR (CDCl3) δ 0.77 (s, CH3-19), 0.83 (s, CH3-18), 0.82-2.00 (unassigned CH and CH2),
2.34 (t, J = 7.0 Hz, NCH2CH2), 2.38 (s, CH2N), 2.62 (t, J = 7.6 Hz, CH2CH2Ph), 3.18 (s,
NCH2C=O), 3.89 (m, OCH2CH2O), 7.18 (m, 3H of Ph), 7.28 (m, 2H of Ph) ppm. 13C NMR
(CDCl3) δ 11.4, 14.4, 20.4, 22.7, 25.9, 28.1, 28.8, 30.7, 31.0, 31.9, 32.9, 34.2, 35.6, 35.8,
36.0, 38.5, 39.6, 46.0, 50.2, 53.4, 55.5, 56.7, 60.4, 64.6, 65.1, 82.3, 119.4, 125.8, 128.3 (2C),
128.4 (2C), 142.2, 168.5 ppm. LR-MS: calcd. for C34H50NO4 [M+H]+ 536.37, found 536.45.

11
2.1.4.15. (3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-[2-(trifluoromethyl)benzyl]
tetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6e10)
Amorphous solid (57 % yield). Rf = 0.8 (hexanes/EtOAc, 1:1). IR (film) ν 1732 (C=O) cm-1.
1H NMR (CDCl3) δ 0.73 (s, CH3-19), 0.82 (s, CH3-18), 0.80-1.98 (unassigned CH and CH2),
2.48 (s, CH2N), 3.28 (d, J = 2.5 Hz, NCH2C=O), 3.67 (s, CH2-Ph), 3.88 (m, OCH2CH2O),
7.39 (tapp, J = 7.6 Hz, 1H of Ar), 7.55 (t, J = 7.5 Hz, 1H of Ar), 7.68 (m, 2H of Ar) ppm.
13CNMR (CDCl3) δ 11.4, 14.4, 20.3, 22.7, 28.1, 30.6, 31.0, 31.8, 32.9, 34.2, 35.7, 36.0, 38.4,
39.5, 45.9, 50.1, 53.4, 55.3, 57.1, 60.1, 64.6, 65.1, 82.4, 119.4, 124.3 (q, JC-F = 273.9 Hz),
126.1 (q, JC-C-C-F = 5.8 Hz), 127.6, 128.9 (q, JC-C-F = 32.8 Hz), 130.4, 132.0, 135.7, 167.9
ppm. LR-MS: calcd. for C32H43F3NO4 [M+H]+ 562.31, found 562.40.
2.1.4.16.
(3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-(naphthalen-2-ylmethyl)
tetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4] oxazinan]-6''-one (6e11)
Amorphous solid (74 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1732 (C=O) cm-1.
1H NMR (CDCl3) δ 0.69 (s, CH3-19), 0.82 (s, CH3-18), 0.80-1.99 (unassigned CH and CH2),
2.44 (s, CH2N), 3.33 (s, NCH2C=O), 3.67 (d, J = 2.8 Hz, CH2-Ph), 3.89 (m, OCH2CH2O),
7.48 (m, 3H), 7.72 (s, 1H of Ar), 7.83 (m, 3H of Ar) ppm. LR-MS: calcd. for C35H46NO4
[M+H]+ 544.33, found 544.45.
2.1.4.17. (3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-[3-(trifluoromethyl)benzyl]
tetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6e12)
Amorphous solid (92 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1732 (C=O) cm-1.
1H NMR (CDCl3) δ 0.73 (s, CH3-19), 0.83 (s, CH3-18), 0.85-2.00 (unassigned CH and CH2),
2.43 (s, CH2N), 3.29 (s, NCH2C=O), 3.57 (s, CH2-Ph), 3.89 (m, OCH2CH2O), 7.52 (m, 4H of
13
Ar) ppm. C NMR (CDCl3) δ 11.4, 14.4, 20.3, 22.6, 28.1, 30.6, 31.0, 31.8, 32.9, 34.2, 35.7,
36.0, 38.4, 39.6, 45.9, 50.1, 53.4, 55.4, 59.8, 60.7, 64.5, 65.1, 82.4, 119.4, 124.0 (q, JC-F
272.3 Hz), 124.5 (q, JC-C-C-F = 3.6 Hz), 125.3 (q, JC-C-C-F = 3.7 Hz), 129.0, 131.0 (q, JC-C-F
=
=
32.3 Hz), 131.9, 137.9, 167.8 ppm. LR-MS: calcd. for C32H43F3NO4 [M+H]+ 562.31, found
562.35.
2.1.4.18.
(3'R,5'S,8'R,9'S,10'S,13'S,14'S)-4''-[3,5-bis(trifluoromethyl)benzyl]-10',13'-
dimethyltetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenan
threne-3',2''-[1,4]oxazinan]-6''-one (6e13)
Amorphous solid (88 % yield). Rf = 0.8 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.73 (s, CH3-19), 0.83 (s, CH3-18), 0.85-2.00 (unassigned CH and CH2),
2.43 (s, CH2N), 3.33 (s, NCH2C=O), 3.65 (s, CH2-Ph), 3.89 (m, OCH2CH2O), 7.82 (s, 3H of
13
Ar) ppm. C NMR (CDCl3) δ 11.3, 14.4, 20.3, 22.6, 28.0, 30.6, 31.0, 31.7, 32.8, 34.2, 35.7,
36.0, 38.5, 39.5, 45.9, 50.1, 53.4, 55.3, 59.7, 60.0, 64.5, 65.1, 82.4, 119.4, 121.7 (q, JC-C-C-F
=
3.7 Hz), 123.2 (q, JC-F = 272.3 Hz) (2C), 128.5 (2C), 132.0 (q, JC-C-F = 33.6 Hz) (2C), 139.8,
167.3 ppm. LR-MS: calcd. for C33H42F6NO4 [M+H]+ 630.29, found 630.30.
2.1.4.19.
(3'R,5'S,8'R,9'S,10'S,13'S,14'S)-4''-[2,4-bis(trifluoromethyl)benzyl]-10',13'-
dimethyltetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenan
threne-3',2''-[1,4]oxazinan]-6''-one (6e14)

12
Amorphous solid (71 % yield). Rf = 0.8 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.74 (s, CH3-19), 0.83 (s, CH3-18), 0.82-2.02 (unassigned CH and CH2),
2.49 (s, CH2N), 3.32 (d, J = 2.0 Hz, NCH2C=O), 3.73 (s, CH2-Ph), 3.89 (m, OCH2CH2O),
7.83 (d, J = 8.2 Hz, 1H of Ar), 7.92 (d, J = 7.5 Hz, 2H of Ar) ppm. 13C NMR (CDCl3) δ 11.4,
14.4, 20.3, 22.6, 28.1, 30.6, 31.0, 31.9, 32.8, 34.2, 35.7, 36.0, 38.4, 39.5, 45.9, 50.1, 53.4,
55.3, 56.6, 60.1, 64.5, 65.1, 82.3, 119.4, 121.9, 123.3 (q, JC-C-C-F = ~3 Hz), 123.4 (q, JC-F
274.1 Hz) (2C), 128.8 (q, JC-C-C-F = 3.2 Hz), 129.5 (q, JC-C-F = 31.1 Hz), 132.2 (q, JC-C-F
=
=
~32.8 Hz), 140.2, 167.4 ppm. LR-MS: calcd. for C33H42F6NO4 [M+H]+ 630.29, found 630.30.
2.1.4.20.
(3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-(naphthalen-1-ylmethyl)
tetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6e15)
Amorphous solid (89 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1732 (C=O) cm-1.
1H NMR (CDCl3) δ 0.68 (s, CH3-19), 0.81 (s, CH3-18), 0.80-2.05 (unassigned CH and CH2),
2.49 and 2.54 (2d of AB system, J = 12.1 Hz, CH2N), 3.25 and 3.32 (2d of AB system, J =
17.4 Hz, NCH2C=O), 3.89 (m, OCH2CH2O), 3.93 (d, J = 3.5 Hz, CH2-Ph), 7.40 (m, 2H of
Ar), 7.51 (m, 2H of Ar), 7.84 (m, 2H of Ar), 8.23 (m, 2H of Ar) ppm. 13C NMR (CDCl3) δ
11.4, 14.4, 20.3, 22.7, 28.1, 30.6, 31.0, 31.8, 32.9, 34.2, 35.7, 36.0, 38.4, 39.5, 45.9, 50.1,
53.4, 55.4, 60.1, 60.1, 64.5, 65.1, 82.5, 119.4, 124.6, 125.1, 125.9 (2C), 127.6, 128.5, 128.7,
132.1, 132.2, 133.9, 168.2 ppm. LR-MS: calcd. for C35H46NO4 [M+H]+ 544.33, found 544.40.
2.1.4.21. (3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-(prop-2-en-1-yl)tetradeca
hydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6e16)
Amorphous solid (86 % yield). Rf = 0.8 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.77 (s, CH3-19), 0.83 (s, CH3-18), 0.75-2.00 (unassigned CH and CH2),
2.44 (s, CH2N), 2.99 (d, J = 6.2 Hz, NCH2CH=CH2), 3.22 (d, J = 2.6 Hz, NCH2C=O), 3.89
(m, OCH2CH2O), 5.22 (m, CH=CH2), 5.78 (m, CH=CH2) ppm. 13C NMR (CDCl3) δ 11.4,
14.4, 20.3, 22.7, 28.1, 30.6, 31.0, 31.9, 32.9, 34.2, 35.7, 36.0, 38.5, 39.6, 45.9, 50.1, 53.4,
55.2, 59.9, 59.9, 64.5, 65.1, 82.4, 118.8, 119.4, 133.6, 168.3 ppm. LR-MS: calcd. for
C27H42NO4 [M+H]+ 444.30, found 444.35.
2.1.4.22.
(3'R,5'S,8'R,9'S,10'S,13'S,14'S)-4''-(4-iodobenzyl)-10',13'-dimethyltetradeca
hydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6e17)
Amorphous solid (93 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1732 (C=O) cm-1.
1H NMR (CDCl3) δ 0.73 (s, CH3-19), 0.82 (s, CH3-18), 0.80-2.00 (unassigned CH and CH2),
2.39 (s, CH2N), 3.26 (s, NCH2C=O), 3.45 (s, CH2-Ph), 3.89 (m, OCH2CH2O), 7.06 (d, J = 8.2
Hz, 2H of Ar), 7.67 (d, J = 8.3 Hz, 2H of Ar) ppm. 13C NMR (CDCl3) δ 11.4, 14.4, 20.3, 22.7,
28.1, 30.6, 31.0, 31.8, 32.9, 34.2, 35.7, 36.0, 38.4, 39.6, 45.9, 50.1, 53.4, 55.4, 59.7, 60.7,
64.5, 65.1, 82.4, 93.0, 119.4, 130.6 (2C), 136.5, 137.7 (2C), 167.9 ppm. LR-MS: calcd. for
C31H43INO4 [M+H]+ 620.22, found 620.25.
2.1.4.23. (3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-(prop-2-yn-1-yl)tetradeca
hydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenanthrene-3',2''-
[1,4]oxazinan]-6''-one (6e18)
To a solution of amine 4e (0.15 mmol) in DMF (4 mL) was added K2CO3 (0.22 mmol)
and a solution of propargyl bromide (0.45 mmol) in toluene (53 µL). The mixture was
refluxed for 17 h and filtered. The DMF was then co-evaporated several times with EtOAc,

13
and the oily crude product was purified by flash chromatography with a mixture of
hexanes/EtOAc/TEA (93:6:1) to give 6e18. Amorphous solid (65 % yield). Rf = 0.4
(hexanes/EtOAc, 7:3). IR (film) ν 1732 (C=O) cm-1. 1H NMR (CDCl3) δ 0.77 (s, CH3-19),
0.82 (s, CH3-18), 0.80-2.04 (unassigned CH and CH2), 2.31 (t, J = 2.2 Hz, CH of alkyne),
2.55 (s, CH2N), 3.35 (s, NCH2C=O), 3.37 (d, J = 2.3 Hz, NCH2CCH), 3.88 (m, OCH2CH2O)
ppm. 13C NMR (CDCl3) δ 11.4, 14.4, 20.3, 22.7, 28.1, 30.6, 31.0, 31.8, 32.9, 34.2, 35.7, 36.0,
38.5, 39.6, 45.2, 45.9, 50.1, 53.0, 53.4, 58.8, 64.5, 65.1, 74.7, 76.5, 82.0, 119.4, 168.1 ppm.
LR-MS: calcd. for C27H40NO4 [M+H]+ 442.29, found 442.35.
2.1.4.24. (3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-{[3'-(trifluoromethyl)biphe
nyl-4-yl]methyl}tetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]
phenanthrene-3',2''-[1,4]oxazinan]-6''-one (6e19)
The tertiary amine 6e17 (0.07 mmol), 3-fluoromethylphenyl boronic acid (0.11 mmol),
[1,1'-bis(diphenylphosphino)ferrocene]-dichloro palladium (II) (8 mol %) and Na2CO3 (0.11
mmol) were mixed in a 3:1 dioxane/water solution (4 mL) and the mixture refluxed under
argon atmosphere for 5h. The reaction mixture was then washed with water and extracted
with EtOAc. The organic layer was dried with anhydrous Na2SO4, filter and the crude
mixture was purified by flash chromatography to give 6e19. Amorphous solid (23 % yield).
1
Rf = 0.2 (hexanes/EtOAc, 7:3). IR (film) ν 1732 (C=O) cm-1. H NMR (CDCl3) δ 0.73 (s,
CH3-19), 0.83 (s, CH3-18), 0.80-2.05 (unassigned CH and CH2), 2.46 (s, CH2N), 3.30 (s,
NCH2C=O), 3.57 (d, J = 2.0 Hz, CH2-Ph), 3.89 (m, OCH2CH2O), 7.42 (d, J = 8.1 Hz, 2H of
Ar), 7.58 (m, 4H of Ar), 7.78 (d, J = 7.5 Hz, 1H of Ar), 7.84 (s, 1H of Ar) ppm. 13C NMR
(CDCl3) δ 11.4, 14.4, 20.3, 22.7, 28.1, 30.6, 31.0, 31.9, 32.9, 34.2, 35.7, 36.0, 38.5, 39.6,
45.9, 50.1, 53.4, 55.5, 59.8, 60.9, 64.6, 65.1, 82.5, 119.4, 123.8 (q, JC-C-C-F = 3.7 Hz), 124.0 (q,
JC-C-C-F = 3.6 Hz), 127.2, 127.3 (2C), 129.3 (2C), 130.3, 131.2 (q, JC-C-F = 32.1 Hz), 136.7,
139.1, 141.5, 149.7, 168.1 ppm. LR-MS: calcd. for C38H47F3NO3 [M+H]+ 638.34, found
638.40.
2.1.4.25.
(3'R,5'S,8'R,9'S,10'S,13'S,14'S)-4''-[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]-
10',13'-dimethyltetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]
phenanthrene-3',2''-[1,4]oxazinan]-6''-one (6e20)
To a solution of amine 6e18 (0.06 mmol) in a 1:1 mixture of n-butanol/water (3 mL)
was added (azidomethyl)benzene (0.11 mmol), a molar solution of cupper (II) sulfate
pentahydrate (63 µL) and a molar solution of sodium ascorbate (125 µL). The reaction
mixture was stirred at room temperature for 3 h, washed with water and extracted with
EtOAc. The organic layer was dried with anhydrous Na2SO4 and filtered. The crude product
was purified by flash chromatography to give 6e20. Amorphous solid (82 % yield). Rf = 0.1
1
(hexanes/EtOAc, 1:1). IR (film) ν 1728 (C=O) cm-1. H NMR (CDCl3) δ 0.71 (s, CH3-19),
0.82 (s, CH3-18), 0.70-2.04 (unassigned CH and CH2), 2.47 (s, CH2N); 3.26 (s, NCH2C=O),
3.67 (s, NCH2C=C-N), 3.88 (m, OCH2CH2O), 5.53 (s, NCH2Ph), 7.27 (s, 1H of triazole), 7.37
(m, 5H of Ph) ppm. 13C NMR (CDCl3) δ 11.4, 14.4, 20.3, 22.6, 28.1, 30.6, 31.0, 31.8, 32.8,
34.2, 35.7, 36.0, 38.4, 39.5, 45.9, 50.1, 51.6, 53.4, 54.2, 54.9, 59.6, 64.5, 65.1, 82.3, 119.4,
122.5, 128.0 (2C), 128.9, 129.2 (2C), 134.5, 167.9 ppm. LR-MS: calcd. for C34H47N4O4
[M+H]+ 575.35, found 575.40.
2.1.4.26.
(3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-{[4-(trifluoromethyl)phe
nyl]sulfonyl}tetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenan
threne-3',2''-[1,4]oxazinan]-6''-one (6e21)

14
To a solution of the amine 4e (0.14 mmol) in DCM was added TEA (0.41 mmol and
the mixture was stirred 5 min under an argon atmosphere. 4-(Trifluoromethyl)
benzenesulfonyl chloride (0.22 mmol) was then added and the solution stirred at room
temperature. After 3 h, DCM is partially evaporated and silica gel added to make a dry-pack
with the crude mixture. The residue was purified by flash chromatography with a mixture of
hexanes/EtOAc/TEA (90:10:1) to give 6e21. Amorphous solid (78% yield). Rf = 0.8
(hexanes/EtOAc, 1:1). IR (film) ν 1728 (C=O) cm-1. 1H NMR (CDCl3) δ 0.79 (s, CH3-19),
0.83 (s, CH3-18), 0.80-2.00 (unassigned CH and CH2), 3.12 (d, J = 2.6 Hz, CH2N), 3.89 (m,
OCH2CH2O and NCH2C=O), 7.85 (d, J = 8.4 Hz, 2H of Ar), 7.91 (d, J = 8.4 Hz, 2H of Ar)
ppm. 13C NMR (CDCl3) δ 11.4, 14.4, 20.4, 22.6, 27.9, 30.6, 30.9, 30.9, 32.6, 34.1, 35.7, 35.8,
37.4, 39.6, 45.9, 46.1, 50.1, 52.1, 53.4, 64.5, 65.2, 82.4, 119.3, 123.0 (q, JC-F = 273 Hz),
126.8 (q, JC-C-C-F = 3.4 Hz) (2C), 128.1 (2C), 135.4 (q, JC-C-F = 33.3 Hz), 138.9, 163.9 ppm.
LR-MS: calcd. for C31H41F3NO6S [M+H]+ 612.25, found 612.35.
2.1.4.27.
(3'R,5'S,8'R,9'S,10'S,13'S,14'S)-10',13'-dimethyl-4''-{[4-(trifluoromethyl)
phenyl]}tetradecahydro-2'H,6''H-dispiro[1,3-dioxolane-2,17'-cyclopenta[a]phenan
threne-3',2''-[1,4]oxazinan]-6''-one (6e22)
This compound was synthesized as reported for 6e21 except that 4-
trifluoromethyl)benzensulfonylchloride was used to give 6e22. Amorphous solid (86 %
yield). Rf = 0.5 (hexanes/EtOAc, 1:1). IR (film) ν 1740 (C=O, morpholinone), 1647 (C=O,
amide) cm-1. 1H NMR (CDCl3) δ 0.82 (s, CH3-19 and CH3-18), 0.75-2.04 (unassigned CH and
CH2), 3.42 and 3.85 (2m, CH2N), 3.89 (m, OCH2CH2O), 4.19 and 4.53 (2m, NCH2C=O), 7.54
(d, J = 8.0 Hz, 2H of Ar), 7.74 (d, J = 8.0 Hz, 2H of Ar) ppm. 13C NMR (CDCl3) δ 11.4, 14.4,
20.4, 22.6, 28.0, 30.5, 30.9, 32.6, 34.1, 35.7, 35.8, 37.1, 39.5, 45.9, 50.1, 53.4, 64.5, 65.1,
82.2, 119.3, 123.5 (q, JC-F = 272.6 Hz), 126.0 (q, JC-C-C-F = 3.5 Hz) (2C), 127.6 (2C), 132.6 (q,
JC-C-F = 35.9 Hz), 137.7, 168.9 (2C) ppm. LR-MS: calcd. for C32H43F3NO4 [M+H]+ 576.29,
found 576.35.
2.1.5. General procedure to generate the spiromorpholinones 5a-5e and the N-
benzylated spiromorpholinones 7a-7e and 7e1-7e22
To a solution of compounds 4a-4e, 6a-6e and 6e1-6e22 (30 - 60 mg) in dioxane (2
mL) was added an aqueous solution of sulfuric acid (5%) (2 mL) and the mixture was stirred
at room temperature (2 h – 5 h). A saturated aqueous solution of Na2CO3 (12 mL) was added
and the reaction mixture extracted with EtOAc. The organic phase was collected, dried with
anhydrous MgSO4 and evaporated under reduced pressure. The purification was performed by
flash column chromatography.
2.1.5.1.
(3R,5S,5'S,8R,9S,10S,13S,14S)-10,13-dimethyl-5'-(2-methylpropyl)tetradeca
hydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (5a)
Amorphous solid (71 % yield). Rf = 0.5 (hexanes/EtOAc, 1:1). IR (film) ν 3448 and 3333
(NH) and 1736 (C=O) cm-1. 1H NMR (acetone-d6) δ 0.84 and 0.85 (2s, CH3-18 and CH3-19),
0.90 and 0.93 (2d, J = 6.7 Hz, (CH3)2- from iPr), 1.05 (m, 1H), 1.10-2.02 (unassigned CH and
CH2), 2.38 (m, CH-16β), 2.82 and 2.95 (2d of AB system, J = 13.4 Hz, CH2N), 3.44 (dd, J1 =
4.1 Hz, J2 = 9.6 Hz, NCHC=O) ppm. 13C NMR (acetone-d6) δ 10.7, 13.2, 20.1, 20.6, 21.4,
22.9, 24.2, 27.9, 30.7, 31.2, 31.7, 33.2, 35.0, 35.1, 36.1, 37.9, 39.8, 41.3, 47.3, 51.3, 51.9,
54.3, 55.5, 81.8, 170.8, 219.0 ppm. LR-MS: calcd. for C26H42NO3 [M+H]+ 416.31, found
416.35.

15
2.1.5.2.
(3R,5S,5'R,8R,9S,10S,13S,14S)-10,13-dimethyl-5'-(2-methylpropyl)tetradeca
hydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (5b)
Amorphous solid (92 % yield). Rf = 0.44 (hexanes/EtOAc, 1:1). IR (film) ν 3448 and 3340
(NH) and 1736 (C=O) cm-1. 1H NMR (CDCl3) δ 0.80 (s, CH3-19), 0.86 (s, CH3-18), 0.93 and
0.97 (2d, J = 6.4 Hz, (CH3)2- from iPr), 0.99-1.97 (unassigned CH and CH2), 2.08 (m, CH-
16α), 2.42 (m, CH-16β), 2.82 and 2.89 (2d of AB system, J = 13.5 Hz, CH2N), 3.50 (dd, J1 =
3.6 Hz and J2 = 9.8 Hz, NCHC=O) ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.0, 21.8,
23.4, 24.5, 28.0, 30.5, 31.4, 31.5, 32.6, 35.0, 35.8, 36.1, 38.2, 39.7, 41.4, 47.8, 51.4, 52.5,
53.8, 55.6, 82.2, 171.8, 221.2 ppm. LR-MS: calcd. for C26H42NO3 [M+H]+ 416.31, found
416.35.
2.1.5.3.
(3R,5S,5'S,8R,9S,10S,13S,14S)-5'-benzyl-10,13-dimethyltetradecahydro-6'H-
spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (5c)
Amorphous solid (97 % yield). Rf = 0.7 (hexanes/EtOAc, 3:7). IR (film) ν 3333 (NH) and
1
1736 (C=O) cm-1. H NMR (acetone-d6) δ 0.81 (s, CH3-19), 0.84 (s, CH3-18), 0.80-1.98
(unassigned CH and CH2), 2.38 (m, CH-16β), 2.79 and 2.90 (2d of AB system, J = 13.3 Hz,
CH2N), 3.02 and 3.16 (2m, CH2Ph), 3.73 (dd, J1 = 4.0 Hz and J2 = 8.1 Hz, NCHC=O), 7.24
(m, 1H of Ph), 7.29 (m, 4H of Ph) ppm. 13C NMR (CDCl3) δ 11.3, 13.8, 20.2, 21.7, 27.8,
30.5, 31.0, 31.5, 32.8, 35.0, 35.8, 36.0, 38.0, 39.2, 47.8, 51.4, 52.6, 53.7, 53.8, 58.6, 58.7,
82.7, 127.1, 128.8 (2C), 129.5 (2C), 137.2, 170.8, 221.3 ppm. LR-MS: calcd. for C29H40NO3
[M+H]+ 450.29, found 450.35.
2.1.5.4. (3R,5S,5'R,8R,9S,10S,13S,14S)-5'-benzyl-10,13-dimethyltetradecahydro-6'H-
spiro [cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (5d)
Amorphous solid (56 % yield). Rf = 0.6 (hexanes/EtOAc, 3:7). IR (film) ν 3448 and 3325
(NH), and 1728 (C=O) cm-1 1H NMR (acetone-d6) δ 0.80 (s, CH3-19), 0.83 (s, CH3-18), 0.80-
.
2.00 (unassigned CH and CH2), 2.37 (m, CH-16β), 2.79 and 2.88 (2d of AB system, J = 13.3
Hz, CH2N), 3.10 (m, CH2Ph), 3.75 (dd, J1 = 4.1 Hz and J2 = 7.5 Hz, NCHC=O), 7.27 (m, Ph)
ppm. 13C NMR (CDCl3) δ 11.3, 13.8, 20.2, 21.8, 27.8, 30.5, 31.3, 31.5, 32.5, 35.0, 35.8, 36.0,
37.7, 38.0, 39.6, 47.8, 51.4, 52.7, 53.8, 58.8, 82.7, 127.1, 128.8 (2C), 129.6 (2C), 137.2,
170.7, 221.2 ppm. LR-MS: calcd. for C29H40NO3 [M+H]+ 450.29, found 450.35.
2.1.5.5. (3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyltetradecahydro-6'H-spiro[cyclopenta
[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (5e)
Amorphous solid (54 % yield). Rf = 0.29 (hexanes/EtOAc, 1:1). IR (film) ν 3448 and 3333
(NH) and 1736 (C=O) cm-1 1H NMR (CDCl3) δ 0.80 (s, CH3-19), 0.86 (s, CH3-18), 0.80-1.99
.
(unassigned CH and CH2), 2.07 (m, CH-16α), 2.43 (dd, J1 = 8.4 Hz, J2 = 19.2 Hz, CH-16β),
2.83 (s, CH2N), 3.63 (s, NCH2C=O) ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.8, 27.9,
30.5, 31.2, 31.5, 32.7, 35.0, 35.8, 36.1, 37.9, 39.4, 47.6, 47.8, 51.4, 52.9, 53.8, 82.5, 168.7,
221.3 ppm. LR-MS: calcd. for C22H34NO3 [M+H]+ 360.25, found 360.25.
2.1.5.6.
(3R,5S,5'S,8R,9S,10S,13S,14S)-4'-benzyl-10,13-dimethyl-5'-(2-methylpropyl)
tetradecahydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-
dione (7a)
Amorphous solid (81 % yield). Rf = 0.77 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1
.
1H NMR (acetone-d6) δ 0.77 (s, CH3-19), 0.83 (s, CH3-18), 0.93 and 0.98 (2d, J = 6.4 Hz,
(CH3)2- from iPr), 0.80-1.99 (unassigned CH and CH2), 2.27 and 2.75 (2d of AB system, J =
12.5 Hz, CH2N), 2.37 (dd, J1 = 8.7 Hz, J2 = 18.3 Hz, CH-16β), 3.17 (dd, J1 = 2.7 and J2 = 6.9
Hz, 1H, NCHC=O), 3.21 and 4.07 (2d of AB system, J = 13.6 Hz, CH2-Ph), 7.34 (m, 1H of

16
Ph), 7.37 (m, 4H of Ph) ppm. 13C NMR (acetone-d6) δ 10.8, 13.2, 20.1, 21.4, 21.6, 23.4, 24.9,
27.8, 30.6, 31.5, 31.7, 33.3, 34.9, 35.1, 36.0, 38.2, 38.3, 39.7, 47.3, 51.3, 54.3, 57.6, 57.7,
63.1, 80.5, 127.2, 128.4 (2C), 128.6 (2C), 138.2, 169.8, 218.7 ppm. LR-MS: calcd. for
C33H48NO3 [M+H]+ 506.36, found 506.40.
2.1.5.7.
(3R,5S,5'R,8R,9S,10S,13S,14S)-4'-benzyl-10,13-dimethyl-5'-(2-methylpropyl)
tetradecahydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-
dione (7b)
Amorphous solid (92 % yield). Rf = 0.85 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1
.
1H NMR (CDCl3) δ 0.71 (s, CH3-19), 0.84 (s, CH3-18), 0.95 and 0.99 (2d, J = 6.6 Hz, (CH3)2-
from iPr), 0.80-2.15 (unassigned CH and CH2), 2.18 and 2.65 (2d of AB system, J = 12.4 Hz,
CH2N), 2.43 (dd, J1 = 8.7 Hz, J2 = 19.2 Hz, CH-16β), 3.12 and 4.03 (2d of AB system, J =
13.5 Hz, CH2-Ph), 3.18 (dd, J1 = 2.9 Hz and J2 = 6.8 Hz, NCHC=O), 7.33 (m, Ph) ppm. 13C
NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.8, 22.1, 24.0, 25.0, 28.1, 30.6, 31.5, 31.9, 32.7, 35.0,
35.8, 36.1, 38.2, 38.5, 39.8, 47.8, 51.4, 53.8, 57.7, 58.1, 63.1, 80.2, 127.4 , 128.5 (2C), 128.6
(2C), 137.7, 171.3, 221.3 ppm. LR-MS: calcd. for C33H48NO3 [M+H]+ 506.36, found 506.40.
2.1.5.8. (3R,5S,5'S,8R,9S,10S,13S,14S)-4',5'-dibenzyl-10,13-dimethyltetradecahydro-6'H-
spiro [cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (7c)
Amorphous solid (70 % yield). Rf = 0.61 (hexanes/EtOAc, 8:2). IR (film) ν 1728 (C=O) cm-1
.
1H NMR (acetone-d6) δ 0.65 (s, CH3-19), 0.80 (s, CH3-18), 0.75-1.99 (unassigned CH and
CH2), 2.22 and 2.59 (2d of AB system, J = 12.5 Hz, CH2N), 2.36 (dd, J1 = 8.7 Hz, J2 = 18.5
Hz, CH-16β), 3.27 and 4.40 (2d of AB system, J = 13.9 Hz, NCH2-Ph), 3.37 (m, CH2Ph);
3.51 (dd, J1 = 2.9 Hz and J2 = 5.1 Hz, NCHC=O), 7.31 (m, 2xPh) ppm. 13C NMR (acetone-d6)
δ 10.7, 13.1, 20.0, 21.4, 27.7, 30.6, 30.6, 31.6, 33.0, 34.9, 35.1, 35.2, 35.8, 37.9, 39.6, 47.3,
51.2, 54.1, 57.4, 57.7, 65.9, 80.5, 126.5, 127.1, 127.8 (2C), 128.4 (2C), 128.5 (2C), 130.4
(2C), 138.0, 138.1, 169.5, 218.7 ppm. LR-MS: calcd. for C36H46NO3 [M+H]+ 540.34, found
540.40.
2.1.5.9.
(3R,5S,5'R,8R,9S,10S,13S,14S)-4',5'-dibenzyl-10,13-dimethyltetradecahydro-
6'H-spiro [cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (7d)
Amorphous solid (81 % yield). Rf = 0.59 (hexanes/EtOAc, 8:2). IR (film) ν 1728 (C=O) cm-1.
1H NMR (acetone-d6) δ 0.65 (s, CH3-19), 0.80 (s, CH3-18), 0.75-1.99 (unassigned CH and
CH2), 2.22 and 2.60 (2d of AB system, J = 12.4 Hz, CH2N), 2.37 (dd, J1 = 8.6 Hz, J2 = 18.4
Hz, CH-16β), 3.29 and 4.42 (2d of AB system, J =13.8 Hz, NCH2-Ph), 3.41 (m, CH2Ph); 3.51
(dd, J1 = 3 Hz and J2 = 5.0 Hz, NCHC=O), 7.30 (m, 10H, 2xPh) ppm. 13C NMR (acetone-d6)
δ 10.7, 13.2, 20.1, 21.4, 27.4, 30.6, 31.2, 31.7, 32.6, 34.9, 35.0, 35.1, 35.8, 37.2, 40.0, 47.3,
51.2, 54.2, 57.4, 57.8, 65.8, 80.5, 126.6, 127.1, 127.8 (2C), 128.4 (2C), 128.6 (2C), 130.5
(2C), 137.9, 138.1, 169.4, 218.7 ppm. LR-MS: calcd. for C36H46NO3 [M+H]+ 540.34, found
540.40.
2.1.5.10. (3R,5S,8R,9S,10S,13S,14S)-4'-benzyl-10,13-dimethyltetradecahydro-6'H-spiro
[cyclopenta [a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (7e)
Amorphous solid (71 % yield). Rf = 0.7 (DCM/MeOH, 39:1). IR (film) ν 1725 (C=O) cm-1.
1H NMR (CDCl3) δ 0.75 (s, CH3-19), 0.85 (s, CH3-18), 0.85-1.99 (unassigned CH and CH2),
2.11 (m, CH-16α), 2.43 (m, CH2N and CH-16β), 3.27 (s, NCH2C=O), 3.50 and 3.54 (2d of
AB system, J = 4.4 Hz, CH2-Ph), 7.33 (m, Ph) ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2,
21.8, 27.9, 30.6, 31.5, 31.8, 32.9, 35.0, 35.8, 36.1, 38.3, 39.6, 47.8, 51.4, 53.8, 55.4, 59.7,

17
61.4, 82.4, 127.6, 128.5 (2C), 128.7 (2C), 136.7, 168.2, 221.3 ppm. LR-MS: calcd. for
C29H40NO3 [M+H]+ 450.29, found 450.35.
2.1.5.11. (3R,5S,8R,9S,10S,13S,14S)-4'-(4-bromobenzyl)-10,13-dimethyltetradecahydro-
6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (7e1)
Amorphous solid (78 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.75 (s, CH3-19), 0.85 (s, CH3-18), 0.84-1.99 (unassigned CH and CH2),
2.07 (m, CH-16α), 2.40 (s, CH2N), 2.42 (dd, J1 = 8.6 Hz, J2 = 19.5 Hz, CH-16β), 3.26 (s,
NCH2C=O), 3.46 (d, J = 2.5 Hz, CH2-Ph), 7.19 (d, J = 8.3 Hz, 2H of Ar), 7.47 (d, J = 8.3 Hz,
2H of Ar) ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.7, 27.9, 30.6, 31.5, 31.8, 32.8, 35.0,
35.8, 36.1, 38.3, 39.6, 47.8, 51.4, 53.8, 55.4, 59.5, 60.6, 82.3, 121.5, 130.3 (2C), 131.7 (2C),
135.8, 167.9, 221.3 ppm. LR-MS: calcd. for C29H3981BrNO3 [M+H]+ 530.20, found 530.15.
2.1.5.12.
(3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-[4-(trifluoromethoxy)benzyl]
tetradecahydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-
dione (7e2)
Amorphous solid (70 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.76 (s, CH3-19), 0.85 (s, CH3-18), 0.85-1.97 (unassigned CH and CH2),
2.05 (m, CH-16α), 2.42 (dd, J1 = 8.6 Hz, J2 = 19.2 Hz, CH-16β); 2.44 (s, CH2N), 3.25 (s,
NCH2C=O), 3.51 (d, J = 3.8 Hz, CH2-Ph), 7.19 (d, J = 8.1 Hz, 2H of Ar), 7.33 (d, J = 8.5 Hz,
2H of Ar) ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.7, 27.9, 30.6, 31.5, 31.8, 32.8, 35.0,
35.8, 36.1, 38.3, 39.6, 47.8, 51.4, 53.8, 55.3, 59.7, 60.5, 82.3, 120.4 (d, JC-F = 257 Hz) 121.0
(2C), 129.9 (2C), 130.3, 135.4, 148.6, 167.9, 221.3 ppm. LR-MS: calcd. for C30H39F3NO4
[M+H]+ 534.28, found 534.15.
2.1.5.13. (3R,5S,8R,9S,10S,13S,14S)-4'-(biphenyl-4-ylmethyl)-10,13-dimethyltetradeca
hydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (7e3)
Amorphous solid (94 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.76 (s, CH3-19), 0.85 (s, CH3-18), 0.80-1.98 (unassigned CH and CH2),
2.05 (m, CH-16α), 2.43 (dd, J1 = 8.6 Hz, J2 = 19.2 Hz, CH-16β), 2.47 (s, CH2N), 3.30 (s,
NCH2C=O), 3.56 (d, J = 3.6 Hz, CH2-Ph), 7.36 (m, 3H of Ar), 7.45 (m, 2H of Ar), 7.60 (m,
13
4H of Ar) ppm. CNMR (CDCl3) δ 11.4, 13.8, 20.2, 21.8, 27.9, 30.6, 31.5, 31.8, 32.9, 35.0,
35.8, 36.1, 38.4, 39.6, 47.8, 51.4, 53.8, 55.5, 59.7, 61.0, 82.4, 127.0 (2C), 127.3 (2C), 127.4,
128.8 (2C), 129.1 (2C), 135.7, 140.5, 140.6, 168.2, 221.3 ppm. LR-MS: calcd. for C35H44NO3
[M+H]+ 526.32, found 526.30.
2.1.5.14. (3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-(2-phenylethyl)tetradecahydro-
6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (7e4)
Amorphous solid (60 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.79 (s, CH3-19), 0.85 (s, CH3-18), 0.75-2.00 (unassigned CH and CH2),
2.08 (m, CH-16α), 2.43 (dd, J1 = 8.6 Hz, J2 = 19.3 Hz, CH-16β), 2.50 (s, CH2N), 2.62 (tapp., J
= 7.5 Hz, CH2-Ph), 2.78 (tapp., J = 7.6 Hz, NCH2CH2Ph), 3.28 (d, J = 1.8 Hz, NCH2C=O),
7.20 (m, 3H of Ph), 7.28 (m, 2H of Ph) ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.8, 27.9,
30.6, 31.5, 31.7, 32.9, 33.2, 35.0, 35.8, 36.1, 38.4, 39.6, 47.8, 51.4, 53.8, 55.3, 58.6, 60.3,
82.2, 126.3, 128.4 (2C), 128.6 (2C), 139.5, 168.3, 221.3 ppm. LR-MS: calcd. for C30H42NO3
[M+H]+ 464.31, found 464.25.

18
2.1.5.15. (3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-(3-phenylpropyl)tetradecahydro-
6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (7e5)
Amorphous solid (51 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.80 (s, CH3-19), 0.86 (s, CH3-18), 0.80-1.96 (unassigned CH and CH2),
2.08 (m, CH-16α), 2.34 (t, J = 7.0 Hz, NCH2CH2CH2Ph), 2.43 (m, CH-16β), 2.44 (s, CH2N),
2.66 (tapp, J = 7.6 Hz, NCH2CH2CH2Ph), 3.21 (s, NCH2C=O), 7.18 (m, 3H of Ph), 7.29 (m,
2H of Ph) ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.8, 27.9, 28.2, 30.6, 31.5, 31.9, 32.9,
33.0, 35.0, 35.8, 36.1, 38.5, 39.6, 47.8, 51.4, 53.8, 55.4, 56.0, 60.1, 82.2, 126.0, 128.4 (4C),
141.6, 168.4, 221.3 ppm. LR-MS: calcd. for C31H44NO3 [M+H]+ 478.32, found 478.25.
2.1.5.16.
(3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-{2-[4-(trifluoromethyl)phenyl]
ethyl}tetradecahydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-
6',17(2H)-dione (7e6)
Amorphous solid (36 % yield). Rf = 0.5 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.79 (s, CH3-19), 0.86 (s, CH3-18), 0.80-2.02 (unassigned CH and CH2),
2.08 (m, CH-16α), 2.43 (dd, J1 = 8.5 Hz, J2 = 19.2 Hz, CH-16β), 2.50 (s, CH2N), 2.64 (tapp., J
= 7.3 Hz, CH2-Ph), 2.84 (tapp., J = 7.4 Hz, NCH2CH2Ph), 3.29 (d, J = 4.1 Hz, NCH2C=O),
7.31 (d, J = 8.0 Hz, 2H of Ar), 7.55 (d, J = 8.1 Hz, 2H of Ar) ppm. 13C NMR (CDCl3) δ 11.4,
13.8, 20.2, 21.7, 27.8, 30.5, 31.5, 31.7, 32.8, 33.0, 35.0, 35.8, 36.1, 38.1, 39.6, 44.6, 47.8,
51.4, 53.8, 55.2, 57.9, 60.3, 82.2, 125.3 (Jc-c-c = 3.6 Hz) (2C), 129.0 (2C), 133.1, 143.6, 167.9,
221.3 ppm. LR-MS: calcd. for C31H41F3NO3 [M+H]+ 532.30, found 532.20.
2.1.5.17. (3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-[4-(trifluoromethyl)benzyl]tetra
decahydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione
(7e7)
Amorphous solid (78 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.76 (s, CH3-19), 0.85 (s, CH3-18), 0.80-2.01 (unassigned CH and CH2),
2.08 (m, CH-16α), 2.42 (m, CH-16β), 2.43 (s, CH2N), 3.28 (s, NCH2C=O), 3.57 (d, J = 2.7
Hz, CH2-Ph), 7.44 (d, J = 8.0 Hz, 2H of Ar), 7.61 (d, J = 8.1 Hz, 2H of Ar) ppm. 13C NMR
(CDCl3) δ 11.4, 13.8, 20.2, 21.7, 27.9, 30.6, 31.5, 31.8, 32.8, 35.0, 35.8, 36.1, 38.3, 39.6,
47.8, 51.4, 53.8, 55.4, 59.7, 60.8, 82.3, 124.0 (q, J = 270.3 Hz), 125.5 (JC-C-C-F = 3.7 Hz) (2C),
128.8 (2C), 130.0 (JC-C-F = 32.3 Hz), 140.9, 167.8, 221.3 ppm. LR-MS: calcd. for
C30H39F3NO3 [M+H]+ 518.28, found 518.25.
2.1.5.18.
(3R,5S,8R,9S,10S,13S,14S)-4'-[4-(benzyloxy)benzyl]-10,13-dimethyltetradeca
hydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (7e8)
Amorphous solid (88 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.75 (s, CH3-19), 0.85 (s, CH3-18), 0.85-1.97 (unassigned CH and CH2),
2.08 (m, CH-16α), 2.41 (s, CH2N), 2.43 (dd, J1 = 8.5 Hz, J2 = 19.2 Hz, CH-16β), 3.24 (s,
NCH2C=O), 3.46 (d, J = 3.8 Hz, CH2-Ph), 5.07 (s, CH2O), 6.95 (d, J = 8.6 Hz, 2H of Ph), 7.21
13
(d, J = 8.6 Hz, 2H of Ph), 7.39 (m, 5H, Ph) ppm. C NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.8,
27.9, 30.6, 31.5, 31.8, 32.9, 35.0, 35.8, 36.1, 38.4, 39.6, 47.8, 51.4, 53.8, 55.4, 59.6, 60.7,
70.1, 82.4, 114.8 (2C), 127.5 (2C), 128.0, 128.6 (2C), 128.9, 129.9 (2C), 136.9, 158.3, 168.3,
221.3 ppm. LR-MS: calcd. for C36H46NO4 [M+H]+ 556.33, found 556.25.
2.1.5.19. (3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-(4-phenylbutyl)tetradecahydro-
6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (7e9)

19
Amorphous solid (77 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.79 (s, CH3-19), 0.86 (s, CH3-18), 0.80-1.97 (unassigned CH and CH2),
2.08 (m, CH-16α), 2.34 (t, J = 7.0 Hz, NCH2CH2CH2CH2Ph), 2.39 (s, CH2N), 2.43 (dd, J1 =
8.7 Hz, J2 = 19.4 Hz, CH-16β), 2.63 (t, J = 7.5 Hz, NCH2CH2CH2Ph), 3.18 (d, J = 1.9 Hz,
NCH2C=O), 7.17 (m, 3H of Ph), 7.28 (m, 2H of Ph) ppm. 13C NMR (CDCl3) δ 11.4, 13.8,
20.2, 21.8, 25.9, 27.9, 28.8, 30.6, 31.5, 31.8, 32.9, 35.0, 35.6, 35.8, 36.1, 38.5, 39.6, 47.8,
51.4, 53.8, 55.5, 56.7, 60.3, 82.2, 125.8, 128.3 (2C), 128.4 (2C), 142.2, 168.4, 221.3 ppm.
LR-MS: calcd. for C32H46NO3 [M+H]+ 492.34, found 492.25.
2.1.5.20. (3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-[2-(trifluoromethyl)benzyl]tetra
decahydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione
(7e10)
Amorphous solid (67 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.76 (s, CH3-19), 0.85 (s, CH3-18), 0.86-2.03 (unassigned CH and CH2),
2.06 (m, CH-16α), 2.43 (dd, J1 = 8.6 Hz, J2 = 19.2 Hz, CH-16β), 2.49 (s, CH2N), 3.29 (d, J =
3.9 Hz, NCH2C=O), 3.68 (s, CH2-Ph), 7.40 (t, J = 7.6 Hz, 1H of Ar), 7.55 (t, J = 7.5 Hz, 1H of
Ar), 7.67 (m, 2H of Ar) ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.8, 27.9, 30.6, 31.5,
31.8, 32.8, 35.0, 35.8, 36.1, 38.3, 39.6, 47.8, 51.4, 53.8, 55.3, 57.1, 60.0, 82.3, 124.3 (q, JC-F
=
273.8 Hz), 126.1 (q, JC-C-C-F = 5.7 Hz), 127.6, 128.9 (q, JC-C-F = 30.3 Hz), 130.5, 132.0, 135.7,
167.8, 221.2 ppm. LR-MS: calcd. for C30H39F3NO3 [M+H]+ 518.28, found 518.50.
2.1.5.21. (3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-(naphthalen-2-ylmethyl)tetradeca
hydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (7e11)
Amorphous solid (86 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.71 (s, CH3-19), 0.84 (s, CH3-18), 0.80-2.00 (unassigned CH and CH2),
2.08 (m, CH-16α), 2.44 (m, CH-16β), 2.45 (s, CH2N), 3.34 (s, NCH2C=O), 3.67 (d, J = 4.3
Hz, CH2-Ph), 7.50 (m, 3H of Ar), 7.71 (s, 1H of Ar), 7.83 (m, 3H of Ar) ppm. 13C NMR
(CDCl3) δ 11.4, 13.8, 20.2, 21.8, 27.9, 30.6, 31.5, 31.8, 32.9, 34.9, 35.8, 36.1, 38.4, 39.6,
47.8, 51.4, 53.8, 55.6, 59.6, 61.5, 82.4, 126.0, 126.3, 126.7, 127.6, 127.7 (2C), 128.4, 133.0,
133.2, 134.2, 168.2, 221.3 ppm. LR-MS: calcd. for C33H42NO3 [M+H]+ 500.31, found 500.20.
2.1.5.22. (3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-[3-(trifluoromethyl)benzyl]tetra
decahydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione
(7e12)
Amorphous solid (79 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.75 (s, CH3-19), 0.84 (s, CH3-18), 0.82-2.00 (unassigned CH and CH2),
2.07 (m, CH-16α), 2.42 (m, CH-16β), 2.43 (s, CH2N), 3.28 (s, NCH2C=O), 3.57 (s, CH2-Ph),
7.52 (m, 4H of Ar) ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.7, 27.8, 30.5, 31.5, 31.7,
32.8, 35.0, 35.8, 36.1, 38.4, 39.6, 47.8, 51.4, 53.8, 55.4, 59.7, 60.7, 82.3, 124.0 (q, JC-F
272.3 Hz), 124.5 (q, JC-C-C-F = 3.6 Hz), 125.3 (q, JC-C-C-F = 3.7 Hz), 129.1, 131.0 (q, JC-C-F
=
=
32.4 Hz), 131.9, 137.9, 167.8, 221.3 ppm. LR-MS: calcd. for C30H39F3NO3 [M+H]+ 518.28,
found 518.50.
2.1.5.23. (3R,5S,8R,9S,10S,13S,14S)-4'-[3,5-bis(trifluoromethyl)benzyl]-10,13-dimethyl
tetradecahydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-
dione (7e13)
Amorphous solid (85 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.75 (s, CH3-19), 0.84 (s, CH3-18), 0.87-1.99 (unassigned CH and CH2),

20
2.07 (m, CH-16α), 2.42 (m, CH-16β), 2.44 (s, CH2N), 3.33 (d, J = 1.7 Hz, NCH2C=O), 3.65
(s, CH2-Ph), 7.81 (s, 3H of Ar) ppm. 13C NMR (CDCl3) δ 11.3, 13.8, 20.2, 21.7, 27.8, 30.5,
31.5, 31.6, 32.8, 35.0, 35.8, 36.1, 38.4, 39.6, 47.8, 51.3, 53.8, 55.3, 59.6, 60.0, 82.3, 121.7 (q,
JC-C-C-F = 3.7 Hz), 123.2 (q, JC-F = 272.7 Hz) (2C), 128.5 (2C), 132.0 (q, JC-C-F = 33.4 Hz)
(2C), 139.8, 167.3, 221.2 ppm. LR-MS: calcd. for C31H38F6NO3 [M+H]+ 586.27, found
586.15.
2.1.5.24. (3R,5S,8R,9S,10S,13S,14S)-4'-[2,4-bis(trifluoromethyl)benzyl]-10,13-dimethyl
tetradecahydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-
dione (7e14)
Amorphous solid (28 % yield). Rf = 0.8 (hexanes/EtOAc, 1:1). IR (film) ν 1740 (C=O) cm-1.
1H NMR (CDCl3) δ 0.77 (s, CH3-19), 0.85 (s, CH3-18), 0.80-1.97 (unassigned CH and CH2),
2.07 (m, CH-16α), 2.43 (dd, J1 = 8.6 Hz, J2 = 19.3 Hz, CH-16β), 2.49 (s, CH2N), 3.32 (d, J =
3.1 Hz, NCH2C=O), 3.74 (s, CH2-Ph), 7.84 (d, J = 8.1 Hz, 1H of Ar), 7.91 (d, J = 8.7 Hz, 1H
13
of Ar), 7.93 (s, 1H of Ar) ppm. C NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.7, 27.9, 30.5, 31.5,
31.8, 32.8, 35.0, 35.8, 36.1, 38.3, 39.6, 47.8, 51.4, 53.8, 55.3, 56.6, 60.0, 82.2, 123.3 (q, Jc-F
=
270.5 Hz), 123.5 (q, Jc-c-c-F = 3.7 Hz), 124.4 (q, JC-F = 276.2 Hz), 124.8, 128.8, 129.6 (q, JC-C-F
= 31.3 Hz), 130.2 (Jc-c-F = 33.1 Hz), 131.0, 140.1, 167.3, 221.2 ppm. LR-MS: calcd. for
C31H38F6NO3 [M+H]+ 586.27, found 586.25.
2.1.5.25. (3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-(naphthalen-1-ylmethyl)tetradeca
hydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (7e15)
Amorphous solid (86 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.69 (s, CH3-19), 0.83 (s, CH3-18), 0.80-1.93 (unassigned CH and CH2),
2.06 (m, CH-16α), 2.41 (dd, J1 = 8.6 Hz, J2 = 19.2 Hz, CH-16β), 2.52 (d, J = 5.5 Hz, CH2N),
3.29 (d, J = 7.5 Hz, NCH2C=O), 3.92 (d, J = 4.8 Hz, CH2-Ph), 7.40 (m, 2H of Ar), 7.51 (m,
2H of Ar), 7.85 (m, 2H of Ar), 8.22 (m, 1H of Ar) ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2,
21.7, 27.8, 30.5, 31.5, 31.7, 32.8, 35.0, 35.8, 36.1, 38.3, 39.6, 47.8, 51.4, 53.8, 55.4, 59.9,
60.1, 82.4, 124.6, 125.1, 125.9 (2C), 127.6, 128.5, 128.8, 132.1, 132.2, 133.9, 168.2, 221.3
ppm. LR-MS: calcd. for C33H42NO3 [M+H]+ 500.31, found 500.20.
2.1.5.26. (3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-(prop-2-en-1-yl)tetradecahydro-
6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (7e16)
Amorphous solid (55 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.79 (s, CH3-19), 0.85 (s, CH3-18), 0.80-2.00 (unassigned CH and CH2),
2.08 (m, CH-16α), 2.43 (m, CH-16β), 2.44 (s, CH2N), 2.99 (d, J = 3.8 Hz, NCH2CH=CH2),
3.21 (d, J = 6.3 Hz, NCH2C=O), 5.21 (m, CH=CH2), 5.76 (m, CH=CH2) ppm. 13C NMR
(CDCl3) δ 11.4, 13.8, 20.2, 21.8, 27.9, 30.6, 31.5, 31.8, 32.9, 35.0, 35.8, 36.1, 38.5, 39.6,
47.8, 51.4, 53.8, 55.2, 59.8, 59.9, 82.3, 118.9, 133.6, 168.3, 221.3 ppm. LR-MS: calcd. for
C25H38NO3 [M+H]+ 400.28, found 400.15.
2.1.5.27.
(3R,5S,8R,9S,10S,13S,14S)-4'-(4-iodobenzyl)-10,13-dimethyltetradecahydro-
6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (7e17)
Amorphous solid (59 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.75 (s, CH3-19), 0.84 (s, CH3-18), 0.84-1.96 (unassigned CH and CH2),
2.07 (m, CH-16α), 2.39 (s, CH2N), 2.42 (dd, J1 = 8.6 Hz, J2 = 19.4 Hz, CH-16β), 3.26 (s,
NCH2C=O), 3.45 (d, J = 2.5 Hz, CH2-Ph), 7.06 (d, J = 8.2 Hz, Ph), 7.67 (d, J = 8.2 Hz, Ph)
ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.7, 27.9, 30.6, 31.5, 31.8, 32.8, 35.0, 35.8, 36.1,

21
38.3, 39.6, 47.8, 51.4, 53.8, 55.4, 59.5, 60.7, 82.3, 93.1, 130.6 (2C), 136.4, 137.7 (2C), 167.9,
221.2 ppm. LR-MS: calcd. for C29H39INO3 [M+H]+ 576.19, found 576.10.
2.1.5.28. (3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-(prop-2-yn-1-yl)tetradecahydro-
6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-6',17(2H)-dione (7e18)
Amorphous solid (67 % yield). Rf = 0.3 (hexanes/EtOAc, 7:3). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.80 (s, CH3-19), 0.86 (s, CH3-18), 0.75-2.04 (unassigned CH and CH2),
2.09 (m, CH-16α), 2.31 (d, J = 2.3 Hz, CH of alkyne), 2.43 (dd, J1 = 8.6 Hz, J2 = 19.2 Hz,
CH-16β), 2.57 (s, CH2N), 3.36 (d, J = 1.0 Hz, NCH2C=O), 3.38 (d, J = 2.4 Hz, NCH2CCH)
ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.8, 27.9, 29.7, 30.6, 31.5, 32.8, 35.0, 35.8, 36.1,
38.4, 39.6, 45.2, 47.8, 51.4, 53.1, 53.8, 58.7, 74.7, 76.5, 82.0, 168.1, 221.3 ppm. LR-MS:
calcd. for C25H36NO3 [M+H]+ 398.26, found 398.15.
2.1.5.29. (3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-{[3'-(trifluoromethyl)biphenyl-4-
yl]methyl}tetradecahydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-
6',17(2H)-dione (7e19)
Amorphous solid (59 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.76 (s, CH3-19), 0.85 (s, CH3-18), 0.85-2.04 (unassigned CH and CH2),
2.09 (m, CH-16α), 2.43 (dd, J1 = 8.6 Hz, J2 = 19.2 Hz, CH-16β), 2.47 (s-broad, CH2N), 3.31
(s, NCH2C=O), 3.58 (d, J = 3.4 Hz, CH2-Ph), 7.42 (d, J = 8.1 Hz, 2H of Ar), 7.59 (m, 4H of
13
Ar), 7.78 (d, J = 7.5 Hz, 1H of Ar), 7.84 (s, 1H of Ar) ppm. C NMR (CDCl3) δ 11.4, 13.8,
20.2, 21.8, 27.9, 30.6, 31.5, 31.8, 32.9, 35.0, 35.8, 36.1, 38.4, 39.6, 47.8, 51.4, 53.8, 55.5,
59.7, 60.9, 82.4, 123.0 (q, Jc-F = 270 Hz), 123.8 (q, JC-C-C-F = 3.9 Hz), 124.0 (q, Jc-c-c-F = 3.9
Hz), 127.3 (2C), 129.3 (2C), 130.3, 131.2 (q, Jc-c-F = 34.0 Hz), 136.7, 139.1, 141.5, 149.7,
168.1, 221.3 ppm. LR-MS: calcd. for C36H43F3NO3 [M+H]+ 594.31, found 594.20.
2.1.5.30.
(3R,5S,8R,9S,10S,13S,14S)-4'-[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]-10,13-
dimethyltetradecahydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-
6',17(2H)-dione (7e20)
Amorphous solid (90 % yield). Rf = 0.6 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O), 1454
1
(C-O) cm-1. H NMR (CDCl3) δ 0.75 (s, CH3-19), 0.86 (s, CH3-18), 0.80-1.97 (unassigned
CH and CH2), 2.06 (m, CH-16α), 2.43 (dd, J1 = 8.6 Hz, J2 = 19.2 Hz, CH-16β), 2.50 (s,
CH2N), 3.27 (s, NCH2C=O), 3.69 (s, NCH2C=C-N), 5.54 (s, NCH2Ph), 7.27 (m, 2H of Ph),
7.34 (m, 4H of Ph) ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.7, 27.8, 30.5, 31.5, 31.7,
32.8, 35.0, 35.8, 36.1, 38.3, 39.6, 47.8, 51.4, 51.6, 53.8, 54.2, 54.9, 59.5, 82.2, 122.5, 128.0
(2C), 128.9, 129.2 (2C), 134.6, 143.6, 167.8, 221.2 ppm. LR-MS: calcd. for C32H43N4O3
[M+H]+ 531.33, found 531.25.
2.1.5.31.
(3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-{[4-(trifluoromethyl)phenyl]
sulfonyl}tetradecahydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-
6',17(2H)-dione (7e21)
Amorphous solid (42 % yield). Rf = 0.7 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O) cm-1.
1H NMR (CDCl3) δ 0.82 (s, CH3-19), 0.86 (s, CH3-18), 0.80-2.03 (unassigned CH and CH2),
2.08 (m, CH-16α), 2.44 (dd, J1 = 8.6 Hz, J2 = 19.3 Hz, CH-16β), 3.10 and 3.18 (2d of AB
system, J = 12.5 Hz, CH2N), 3.81 and 3.90 (2d of AB system, J = 17.5 Hz, NCH2C=O), 7.87
and 7.91 (2d of AB system, J = 8.2 Hz, 4H of Ar) ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2,
21.7, 27.7, 30.5, 30.8, 31.5, 32.6, 35.0, 35.8, 36.0, 37.4, 39.6, 46.1, 47.7, 51.3, 52.0, 53.7,
82.3, 124.3 (q, JC-F = 273 Hz), 126.8 (q, JC-C-C-F = 3.4 Hz) (2C), 128.1 (2C), 135.4 (q, JC-C-F
=

22
33.4 Hz), 138.9, 163.8, 221.0 ppm. LR-MS: calcd. for C29H37F3NO5S [M+H]+ 568.23, found
568.10.
2.1.5.32.
(3R,5S,8R,9S,10S,13S,14S)-10,13-dimethyl-4'-{[4-(trifluoromethyl)phenyl]
carbonyl}tetradecahydro-6'H-spiro[cyclopenta[a]phenanthrene-3,2'-[1,4]oxazinane]-
6',17(2H)-dione (7e22)
Amorphous solid (83 % yield). Rf = 0.4 (hexanes/EtOAc, 1:1). IR (film) ν 1736 (C=O), 1647
1
(C=O, amide) cm-1. H NMR (CDCl3) δ 0.85 (s-broad, CH3-19 and CH3-18), 0.80-1.96
(unassigned CH and CH2), 2.07 (m, CH-16α), 2.43 (dd, J1 = 8.7 Hz, J2 = 19.3 Hz, CH-16β),
3.44 and 3.84 (2m, CH2N), 4.20 and 4.53 (2m, NCH2C=O), 7.54 (d, J = 8.0 Hz, 2H of Ar),
7.74 (d, J = 8.1 Hz, 2H of Ar) ppm. 13C NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.7, 27.8, 30.5,
31.4, 32.5, 34.9, 35.8, 35.9, 36.9, 39.5, 47.7, 51.3, 53.7, 57.8, 64.5, 66.1, 123.5 (q, JC-F = 273
Hz), 126.0 (q, JC-C-C-F = 3.5 Hz) (2C), 127.6 (2C), 132.8 (q, JC-C-F = 35.9 Hz), 137.7, 168.9,
221.2 ppm. LR-MS: calcd. for C30H37F3NO4 [M+H]+ 532.26, found 532.20.
2.1.6. Synthesis of carbamates 9a-9e
To a solution of the amino alcohol 3a, 3b, 3c, 3d or 3e (0.12 mmol) in DCM (3 mL)
was added DIPEA (0.24 mmol) at 0 °C, and the solution was stirred under an argon
atmosphere for 10 min. Triphosgene (0.06 mmol) was added and the reaction mixture stirred
for 5 h at room temperature. An acidic solution (2 mL) of HCl/MeOH (10:90) was then added
to the reaction mixture which was stirred overnight at room temperature. The reaction was
stopped by adding a saturated NaHCO3 aqueous solution (10 mL) and the product extracted
with DCM. The organic layer was dried using anhydrous Na2SO4 and evaporated under
reduced pressure. The purification was done by flash chromatography, using
hexanes/EtOAc/TEA (95:5:1) for the elution.
2.1.6.1. Methyl (2S)-2-[(3R,8R,9S,10S,13S,14S)-10,13-dimethyl-2',17-dioxohexadeca
hydro-3'H-spiro[cyclopenta[a]phenanthrene-3,5'-[1,3]oxazolidin]-3'-yl]-4-methyl
pentanoate (9a)
Amorphous solid (73 % yield). Rf = 0.71 (hexanes/EtOAc, 1:1). IR (film) ν 1744
1
(C=O) cm-1. H NMR (CDCl3) δ 0.82 (s, CH3-19), 0.85 (s, CH3-18), 0.96 and 0.97 (2d, J =
1.9 Hz, (CH3)2- from iPr), 0.99-1.97 (unassigned CH and CH2), 2.11 (m, CH-16α), 2.43 (dd,
J1 = 8.4 Hz, J2 = 19.3 Hz, CH-16β), 3.14 and 3.42 (2d of AB system, J = 8.1 Hz, CH2N), 3.72
13
(s, OCH3), 4.58 (dd, J1 = 4.8 Hz and J2 = 11.1 Hz, NCHCO) ppm. CNMR (CDCl3) δ 11.4,
13.8, 20.2, 21.1, 21.8, 23.2, 24.9, 27.9, 30.6, 31.5, 32.7, 33.8, 35.0, 35.4, 35.8, 37.7, 39.5,
40.8, 47.7, 51.3, 52.2, 52.9, 53.6, 53.9, 79.6, 157.6, 172.0, 221.2 ppm. LR-MS: calcd. for
C28H44NO5 [M+H]+ 474.31, found 474.35.
2.1.6.2. Methyl (2R)-2-[(3R,8R,9S,10S,13S,14S)-10,13-dimethyl-2',17-dioxohexadeca
hydro-3'H-spiro[cyclopenta[a]phenanthrene-3,5'-[1,3]oxazolidin]-3'-yl]-4-methyl
pentanoate (9b)28
Amorphous solid (67 % yield). Rf = 0.57 (hexanes/EtOAc, 1:1). IR (film) ν 1736
1
(C=O) cm-1. H NMR (CDCl3) δ 0.83 (s, CH3-19), 0.86 (s, CH3-18), 0.97 (d, J = 6.5 Hz,
(CH3)2- from iPr), 0.80-1.97 (unassigned CH and CH2), 2.05 (m, CH-16α), 2.44 (dd, J1 = 8.4
Hz, J2 = 19.3 Hz, CH-16β), 3.14 and 3.44 (2d of AB system, J = 8.1 Hz, CH2N), 3.72 (s,
OCH3), 4.60 (dd, J1 = 4.8 Hz and J2 = 11.1 Hz, NCHCO) ppm. 13C NMR (CDCl3) δ 11.4,
13.8, 20.2, 21.1, 21.7, 23.1, 24.9, 27.8, 30.6, 31.5, 32.9, 33.9, 35.0, 35.4, 35.8, 37.7, 39.3,
40.8, 47.7, 51.3, 52.2, 52.8, 53.6, 53.9, 79.6, 157.5, 172.0, 221.1 ppm. LR-MS: calcd. for
C28H44NO5 [M+H]+ 474.31, found 474.35.

23
2.1.6.3. Methyl (2S)-2-[(3R,8R,9S,10S,13S,14S)-10,13-dimethyl-2',17-dioxohexadeca
hydro-3'H-spiro[cyclopenta[a]phenanthrene-3,5'-[1,3]oxazolidin]-3'-yl]-3-phenyl
propanoate (9c)
Amorphous solid (61 % yield). Rf = 0.40 (hexanes/EtOAc, 1:1). IR (film) ν 1744
(C=O) cm-1. 1H NMR (CDCl3) δ 0.76 (s, CH3-19), 0.84 (s, CH3-18), 0.75-1.95 (unassigned
CH and CH2), 2.06 (m, CH-16α), 2.42 (dd, J1 = 8.5 Hz, J2 = 19.3 Hz, CH-16β), 2.95 and 3.35
(2m, CH2Ph), 3.09 and 3.33 (2d of AB system, J = 8.0 Hz, CH2N), 3.76 (s, OCH3), 4.88 (dd,
J1 = 5.5 Hz and J2 = 11.3 Hz, NCHCO), 7.22 (m, 3H of Ph), 7.30 (m, 2H of Ph) ppm. 13C
NMR (CDCl3) δ 11.3, 13.8, 20.2, 21.7, 27.7, 30.5, 31.5, 32.4, 33.8, 35.0, 35.1, 35.3, 35.8,
38.8, 40.6, 47.7, 51.3, 52.4, 53.2, 53.8, 55.8, 79.7, 127.1, 128.6 (2C), 128.7 (2C), 136.0,
157.3, 171.0, 221.2 ppm. LR-MS: calcd. for C31H42NO5 [M+H]+ 508.30, found 508.35.
2.1.6.4. Methyl (2R)-2-[(3R,8R,9S,10S,13S,14S)-10,13-dimethyl-2',17-dioxohexadeca
hydro-3'H-spiro[cyclopenta[a]phenanthrene-3,5'-[1,3]oxazolidin]-3'-yl]-3-phenyl
propanoate (9d)
Amorphous solid (75 % yield). Rf = 0.50 (hexanes/EtOAc, 1:1). IR (film) ν 1736
1
(C=O) cm-1. H NMR (CDCl3) δ 0.77 (s, CH3-19), 0.84 (s, CH3-18), 0.75-1.96 (unassigned
CH and CH2), 2.07 (m, CH-16α), 2.42 (dd, J1 = 8.5 Hz, J2 = 19.3 Hz, CH-16β), 2.96 and 3.35
(2m, CH2Ph), 3.10 and 3.33 (2d of AB system, J = 8.0 Hz, CH2N), 3.76 (s, OCH3), 4.87 (dd,
J1 = 5.4 Hz and J2 = 11.2 Hz, 1H, NCHCO), 7.22 (m, 3H of Ph), 7.29 (m, 2H of Ph), ppm. 13C
NMR (CDCl3) δ 11.4, 13.8, 20.2, 21.7, 27.7, 30.5, 31.5, 32.4, 33.7, 35.0, 35.1, 35.3, 35.8,
39.1, 40.8, 47.7, 51.3, 52.4, 53.3, 53.8, 55.9, 79.7, 127.1, 128.5 (2C), 128.7 (2C), 136.0,
157.3, 171.0, 221.2 ppm. LR-MS: calcd. for C31H42NO5 [M+H]+ 508.30, found 508.40.
2.1.6.5. Methyl [(3R,8R,9S,10S,13S,14S)-10,13-dimethyl-2',17-dioxohexadecahydro-3'H-
spiro[cyclopenta[a]phenanthrene-3,5'-[1,3]oxazolidin]-3'-yl]acetate (9e)
Amorphous solid (32 % yield). Rf = 0.48 (hexanes/EtOAc, 1:1). IR (film) ν 1736
1
(C=O) cm-1. H NMR (CDCl3) δ 0.82 (s, CH3-19), 0.85 (s, CH3-18), 0.80-1.97 (unassigned
CH and CH2), 2.10 (m, CH-16α), 2.43 (dd, J1 = 8.4 Hz, J2 = 19.3 Hz, CH-16β), 3.33 (s,
13
CH2N), 3.75 (s, OCH3), 3.99 and 4.04 (2d, J = 18.0 Hz, NCHCO) ppm. C NMR (CDCl3) δ
11.4, 13.8, 20.2, 21.7, 27.8, 30.6, 31.5, 32.7, 33.8, 35.0, 35.4, 35.8, 39.3, 40.8, 45.0, 47.7,
51.3, 52.3, 53.9, 56.5, 79.6, 157.6, 169.0, 221.2 ppm. LR-MS: calcd. for C24H36NO5 [M+H]+
418.25, found 418.30.
2.2. Biological evaluation
2.2.1. Inhibition of 17β-HSD3 (microsomal fraction of rat testes)
A microsomal preparation of rat testes was obtained using a previously described, but
slightly modified procedure.31 In brief, rat testes were homogenized on ice with a Polytron in
cold phosphate buffer (20 mM KH2PO4, 0.25 M sucrose, 1 mM EDTA, pH 7.5) containing
protease inhibitors mini-complete (Roche Diagnostics, Laval, QC, Canada) and the mixture
was centrifuged at 12,500g for 15 min to remove the mitochondria, plasma membranes, and
cell fragments. The supernatant was further centrifuged at 100,000g for 45 min using an
ultracentrifuge equipped with a 70.1 Ti rotor. The microsomal pellet was washed three times
with phosphate buffer and centrifuged at 100,000g for 15 min. All these operations were
conducted at 4 °C. The protein concentration of the supernatant was determined by the
Bradford method using bovine serum albumin as standard. The enzymatic assay was

24
performed at 37 °C for 2 h in 1 mL of a solution containing 860 µL of 50 mM sodium
phosphate buffer (pH 7.4, 20 % glycerol and 1 mM EDTA), 100 µL of 5 mM NADPH in
phosphate buffer, 10 µL of 5 µM [14C]-4-androstene-3,17-dione in ethanol (53.6 mCi/mmol,
Perkin Elmer Life Sciences Inc., Boston, MA, USA), 10 µL of inhibitor dissolved in DMSO
and 20 µL of diluted enzymatic source in phosphate buffer. Each inhibitor was assessed in
triplicate. Afterwards, radiolabelled steroids were extracted from the reaction mixture with
diethyl ether. The organic phase was evaporated to dryness with nitrogen stream. Residue was
dissolved in 50 µL of DCM and dropped on silica gel 60 F254 thin layer chromatography
plates (EMD Chemicals Inc., Gibbstown, NJ, USA) and eluted with a mixture of
toluene/acetone (4:1) as solvent system. Substrate ([14C]-4-dione) and metabolite ([14C]-T)
were identified by comparison with reference steroids and quantified using the Storm 860
System (Molecular Dynamics, Sunnyvale, CA, USA). The percentage of transformation and
then the percentage of inhibition were calculated.
2.2.2. Inhibition of 17β-HSD3 (intact LNCaP cells overexpressing 17β-HSD3)
LNCaP transfected cells (LNCaP[17β-HSD3]) kindly provided by IPSEN
INNOVATION (France) were maintained at 37 °C under 5 % CO2 humidified atmosphere.
Cells were grown in RPMI-1640 medium supplemented (v/v) with 10% fetal bovine serum
(FBS), 1 % penicillin/streptomycin, 2 mM L-glutamine, 4.5 g/L D-glucose, 10 mM Hépès, 1
mM sodium pyruvate and 250 µg/mL hygromycin. For enzymatic assays, the protocol
medium had the same composition, but hygromycin, used to maintain the clone selection, was
not included. LNCaP[17β-HSD3] cells were plated in 24-well culture at 15,000 cells per well,
in protocol medium. After incubation for 3 days, 15 nM of [14C]-4-androstene-3,17-dione and
10 μL of inhibitor dissolved in DMSO were added. The final DMSO concentration in each
well was adjusted to 0.05 %. After 3 h, the culture medium was removed from wells. Steroids
were extracted, separated by TLC, quantified, and percentage of inhibition calculated as
described earlier for enzymatic assay with microsomal fraction of rat testes.
2.2.3. Proliferative activities on LAPC-4 (AR+) cells
Androgen-sensitive human prostate cancer LAPC-4 cells were kindly provided by
Robert E. Reiter from University of California (Los Angeles, CA, USA) and maintained at
37 °C under 5 % CO2 humidified atmosphere. Cells were grown in RPMI-1640 medium
supplemented (v/v) with 10 % fetal bovine serum (FBS), 1 % L-glutamine, 1 % insulin and 1
% penicillin/streptomycin. To determine the effect of novel compounds on cell proliferation,
LAPC-4 cells were suspended with the medium supplemented with 5 % dextran-coated
charcoal treated FBS rather than 10 % FBS to remove the remaining hormones. Triplicate
cultures of 10,000 cells in a total of 100 μL medium in 96-well microtiter plates (Becton–
Dickinson Company, Lincoln Park, NJ, USA) were pre incubated for 24 h at 37 °C under 5 %
CO2 humidified atmosphere. Tested compounds were dissolved in DMSO to prepare the stock
solution of 10−2 M, the compounds were then diluted at several concentrations with culture
medium, added to corresponding wells and incubated for 3 days. The final DMSO
concentration in each well was adjusted to 0.05 %. Control wells were treated with vehicle
DMSO. Quantification of cell growth was determined by MTS method, using CellTitter 96®
AQueous Solution Cell Proliferation Assay (Promega, Nepean, ON, Canada) and following
the manufacturer’s instructions. The proliferative (androgenic) activity was expressed as
difference between the cell proliferation (in %) caused by a given compound and the basal
cell proliferation fixed at 100 %.

25
Scheme 1: Partial numbering for compound 1 and synthesis of target compounds 5a-5e, 7a-7e and 9a-9e.
Reagents and conditions: i. HOCH2CH2OH, p-TSA, toluene, reflux, overnight; ii. NMO, molecular sieves,
TPAP, DCM, rt, 3 h; iii. (CH3)3SOI, NaH, DMSO/THF, rt; iv. Methyl ester of L- or D-Leucine, L- or D-
phenylalanine or glycine methyl esters, MeOH, 90 °C, overnight; v. NaOMe, THF, rt, 2 h; vi. DIPEA,
C6H5CH2Br, DCM, 75 °C, 22 h; vii. H2O:H2SO4, 1,4-dioxane, rt, 2 h to 5 h; viii. (Cl3CO)2CO; DCM; 0 °C to rt, 5
h; ix. HCl:MeOH (1:9), overnight.

26
3. Results and discussion
3.1. Chemical synthesis
3.1.1. Chemical synthesis of steroidal compounds (1st Series, Schemes 1 and 2)
The oxirane 2 (3α-O) was obtained from a well-known procedure already described in
previous work (Scheme 1).26 Briefly, the C-17 ketone of ADT or epi-ADT (1) was protected
as a dioxolane group, the C-3 alcohol was oxidized and the corresponding ketone transformed
to oxirane 2. The inverse stereoisomer (3β-O) was the minor product isolated from the major
product (stereoisomer 3α-O) after recrystallization in acetone. Aminolysis of oxirane 2 in
methanol yielded the amino alcohols 3a-3e with good to excellent yields (53-99 %). A similar
aminolysis was previously done with other amino acids at position C-17 of an estrane nucleus
by Rouillard et al.,30 but in this work, since glycine methyl ester has high polarity, the authors
failed to isolate the free amino acid and to use it for the aminolysis of the C-17-oxirane. To
overcome this problem, we generated glycine methyl ester in situ from its hydrochloride salt,
and the epoxide ring was opened successfully to form the glycine derivative 3e in 53 % yield.
Table 1: Optimization of the lactonization process
Entry Substratea
Base
Base
Equiv.
THF
T
Crude mass
isolated
Total mass of
Yield of
R/S-
R/S-isomers ratiob
(mL) (°C)
pure R/S-isomers
(mg)
(mg)
(%)
NaH
NaH
0.9
1.1
1.1
1.1
1.1
2.2
3.3
<1.1
1.1
0.5
0.6
0.5
1.5
5
22
22
22
22
22
22
22
22
22
0
129
17
17
[53:4
[54:4
[49:5
[66:3
[52:4
----e
1
2
3
4
5
6
7
8
9
3d
11
11
11
11
11
11
11
11
11
11
11
50
66
17
12
24
t-BuOK
NaHMD
LDAc
5
17
5
55
25
36
5
73
50
71
LDAc
5
----e
----e
70
----e
----e
23
----e
----e
33
LDAc
5
----e
LDAd
5
[86:1
[73:2
[77:2
[80:2
[76:2
NaOMe
NaOMe
NaOMe
NaOMe
5
83
63
90
5
82
69
98
10
11
12
12
12
22
-10
85
84f
69
98
42
60
(a) We used 114 mg of starting 3α-OH-amino alcohol 3d (Scheme 1) for entry 1 and 75 mg of 3β-OH-amino
alcohol 11 (Scheme 2) for entries 2-12. (b) Two spots with variable importance on TLC plates. The ratio was
1
calculated from H NMR considering the integration of C-18 singlet. (c) LDA was generated separately in a
large scale. (d) LDA was generated in situ in a small scale (345 μL total volume of solution). (e) Products were
formed but degraded readily; TLC showed total degradation after 4 h of reaction time and no further work-up
was done. (f) 25 % of starting amino alcohol was observed.
The spiromorpholinones 4a-4e were prepared by the intramolecular transesterification
of the amino alcohols 3a-3e, respectively. In our first assay with 0.9 equivalent of sodium

27
hydride (Table 1, entry 1), we observed a poor overall reaction yield (17 %), probably due to
undesired intermolecular side reactions. TLC analysis showed two spots for two major
products, while 1H NMR characterization showed fingerprints for both isolated products, with
a very slight difference on the chemical shift of HC-α, CH2N, CH2Ph and 18-CH3, suggesting
that a racemization occurred. In fact, since the HC-α of the lactone ring is slightly acid,
especially when the geminal group is a benzyl, racemization can occur in the presence of NaH
used as a base. In order to optimize this crucial step and to reduce racemization, several
cyclisation conditions were performed (Table 1, entries 2-12) using the amino alcohol 11,
obtained from the minor oxirane 10 (isomer 3β-O) (Scheme 2). The mixture of the two major
products formed were isolated, purified by chromatography to give the yield, and the R/S-
1
ratio of both isomers was measured from the integration of the H NMR spectra, which
showed two distinctive singlets for 18-CH3 (Fig. 2A). Yield and R/S-ratio were not
significantly improved using the same base (NaH) in a more diluted solution (entry 2). In
order to investigate the effect of a more hindered base on the yield and R/S-ratio, we used
potassium tert-butoxide (KOt-Bu), sodium bis(trimethylsilyl)amide (NaHMDS) and lithium
diisopropylamide (LDA).
Scheme 2: Chemical synthesis of 11 and 12 from oxirane 10. Reagents and conditions: i. D-Phenylalanine,
MeOH, 90 °C, overnight; ii. NaH, KOt-Bu, NaHMDS, LDA or NaOMe, THF, -10 °C, 0 °C or 22 °C, 2 h (see
Table 1).
Figure 2: Fragments of 1H NMR spectra showing that racemization occurs during the lactonization of 11 to 12.
Distinctive 18-CH3 peaks of the two isomers (R and S) were used to calculate the R/S-ratio of the isomeric
mixture; (A) [52:48] and (B) [77:23] (entries 5 and 10, respectively, Table 1).
KOt-Bu caused a total racemization with only 17 % yield of both isomers (entry 3).
Although the global yield was improved with NaHMDS ([66:34] R/S-ratio) and LDA ([52:48]
R/S-ratio) when using these bases in the same conditions (36 % and 71 %, respectively)

28
(entries 4 and 5), the R/S-ratio did not increase. In fact a drastic loss of global yield was
observed with 2.2 and 3.3 equivalents of LDA (entries 6 and 7), whereas a moderate yield of
33 % and a very good R/S-ratio were obtained with less than 1 equivalent of LDA (entry 8).
Sodium methoxide (NaOMe) gave the best yield and least racemization (entries 9-12, Fig.
2B). Among all conditions tested, NaOMe (0.6 equivalent) used at 22 °C in 12 mL of THF
(entry 11) was selected for the lactonization of 3a-3e providing 4a-4e. The high volume of
THF and low quantity of base favor the intramolecular transesterification and avoid undesired
intermolecular side reactions, thus providing the best yields of spiromorpholinones.
The N-benzylated spiromorpholinones 6a-6e were obtained by a nucleophilic
substitution of the secondary amines 4a-4e. Finally, the C-17-dioxolane group of secondary
amines 4a-4e and tertiary amines 6a-6e were hydrolyzed to provide the final compounds 5a-
5e and 7a-7e. The spirocarbamates 8a-8e were synthesized from the amino alcohols 3a-3e,
respectively, following a treatment with triphosgene.26 Intermediates 8a-8e were not isolated
from the reaction mixture and their C-17-dioxolane group was directly hydrolyzed to obtain
9a-9e in 32 to 73 % yields.
3.1.2. Chemical synthesis of steroidal compounds (2nd Series, Scheme 3)
For our SAR study, different benzyl derivatives were also used to introduce molecular
diversity on the spiromorpholinone ring. Starting from compound 4e, a series of tertiary
amines 6e1-6e18 were obtained by nucleophilic substitution (Scheme 3). We also investigated
the Suzuki-Miyaura coupling32 as an additional strategy to add diversity. In fact, when
compound 6e17 was treated with 3-(trifluoromethyl)benzeneboronic acid, no reaction was
observed whith Pd(OAc)2 as catalyst, but using 1,1'-bis(diphenylphosphino)ferrocene-
palladium(II)dichloride (Pd(dppf)Cl2) yielded the biaryl compound 6e19 in 23 % yield. This
poor yield was probably due to the opening of the spirocycle E in aqueous Na2CO3 solution,
but this reaction could be optimized. Starting from 6e18 we also experienced the “click
chemistry” as a mean to diversify the N-substituent. Thus, the steroidal triazolic compound
6e20 was obtained by 1,3-dipolar cycloaddition of the terminal alkyne 6e18 with benzyl
azide.33 It is well known that the 1,4-triazole regioisomer is exclusively obtained from
catalysis with copper (I).34-36 The sulfonamide 6e21 and the carboxamide 6e22 were also
synthesized by nucleophilic substitution under conditions depicted in Scheme 3 and using
benzyl sulfonyl chloride and benzoyl chloride, respectively. The C-17-dioxolane group of the
tertiary amines 6e1-6e22 was finally hydrolyzed to generate the corresponding 17-keto
products 7e1-7e22.

29
Scheme 3: Reagents and conditions. i. DIPEA, X-Ph-CH2Br, DCM, 65-75 °C, 22h; ii. K2CO3, propargyl
bromide, DMF, reflux, overnight; iii. TEA, 4-(trifluoromethyl)benzenesulfonylchloride, DCM, rt, 3 h; iv. TEA,
4-(trifluoromethyl)benzoylchloride, DCM, rt, 3 h; v. 3-(Trifluoromethyl)phenylboronic acid, Pd(dppf)Cl2,
Na2CO3, H2O:1,4-dioxane (3:1), 110 °C, 5 h; vi. Benzyl azide, aqueous CuSO4.5H2O, sodium ascorbate, n-
BuOH:H2O (1:1), rt, 3 h; vii. H2O:H2SO4 (95:5), 1,4-dioxane, rt, 2 to 5 h.
3.1.3. Chemical synthesis of non-steroidal compounds (Scheme 4)
In order to verify the importance of the steroid scaffold for the inhibition of 17β-
HSD3, non-steroidal spiromorpholinones 14, 15 and 17 were synthesized starting from
cyclohexanone and following the sequence of reactions depicted in Scheme 4. Compound 14
was also used to prepare its propargylic derivative 16, which was submitted to a 1,3-dipolar
cycloaddition, under the same conditions used for the preparation of compound 6e20, to
generate compound 17. The preparation of 17 using “click chemistry” represents an
interesting additional strategy to the nucleophilic substitution for introducing molecular
diversity on the spiromorpholinone ring.