Novel Potent Na+,K+-ATPase Inhibitors
J ournal of Medicinal Chemistry, 2003, Vol. 46, No. 17 3653
(E)-4r-Meth yl-3-(2-am in oeth oxyim in o)an dr ostan e-6,17-
d ion e h yd r och lor id e (22d ). Prepared in 60% yield from 1e
and 2-aminoethoxyamine dihydrochloride. The crude product
was crystallized from MeOH/EtOAc. 1H NMR (DMSO-d6) δ
0.77 (s, 3H, CH3), 0.78 (s, 3H, CH3), 0.98 (d, 3H, CH3 eq), 2.60
(dq, 1H, 4-H ax), 3.03 (m, 2H, NCH2), 3.12 (m, 1H, 2-H eq),
4.12 (m, 2H, OCH2), 8.12 (bb, 3H, NH3+); white solid: mp 226-
229 °C dec. Anal. (C22H34N2O3‚HCl) C, H, N, Cl.
(E,Z)-3-(2-Aceta m id oeth oxyim in o)a n d r osta n e-6,17-d i-
on e (22e). To a stirred solution of 22b (1.14 g, 2.9 mmol) in
CH2Cl2 (11.5 mL) at room temperature, Ac2O (0.54 mL, 5.7
mmol) and pyridine (0.46 mL, 5.7 mmol) were added. The
solution was stirred at room temperature for 4 h and evapo-
rated. The residue was dissolved with EtOAc (100 mL), washed
with 1 N HCl (1 × 20 mL) and brine (4 × 20 mL). The organic
solution was dried over Na2SO4, filtered, and evaporated to
dryness. The residue was purified by flash chromatography
(cyclohexane/CH2Cl2/acetone 2/4/4) to give 22e (0.80 g, 69%).
1H NMR (DMSO-d6) δ 0.79 (s, 3H, CH3), 0.81 (s, 3H, CH3),
1.76 (s, 1.5H, COCH3), 1.77 (s, 1.5H, COCH3) 2.96 (m, 0.5H,
4-H eq Z isomer), 3.03 (m, 0.5H, 2-H eq E isomer), 3.22 (m,
2H, NCH2), 3.88 (m, 2H, OCH2), 7.88 (bb, 1H, NH); white
solid: mp 101-103 °C. Anal. (C23H34N2O4‚0.5 H2O) C, H, N,
H2O.
(E ,Z)-3-(3-Am in op r op oxyim in o)a n d r ost a n e -6,17-d i-
on e h yd r och lor id e (22f). Prepared in 73% yield from 1a and
3-aminopropoxyamine dihydrochloride. The crude product was
crystallized from MeOH/EtOAc. 1H NMR (DMSO-d6) δ 0.74
(s, 3H, CH3), 0.75 (s, 3H, CH3), 2.81 (m, 2H, NCH2), 2.95 (m,
0.5H, 4-H eq Z isomer), 3.00 (m, 0.5H, 2-H eq E isomer), 3.99
(m, 2H, OCH2), 7.85 (bb, 3H, NH3+); white solid: mp 83.5-
139.5 °C. Anal. (C22H34N2O3‚HCl) C, H, N, Cl.
(E,Z)-3-(2-Am in oeth oxyim in o)-6r-h yd r oxya n d r osta n e-
17-on e h yd r och lor id e (22g). Prepared in 70% yield from 1b
and 2-aminoethoxyamine dihydrochloride. The crude product
was crystallized from MeOH/EtOAc. 1H NMR (DMSO-d6) δ
0.77 (s, 3H, CH3), 0.86 (s, 1.5H, CH3), 0.87 (s, 1.5H, CH3), 3.02
(t, 2H, NCH2), 3.06 (m, 0.5H, 2-H eq. E isomer), 3.25 (m, 1H,
6-H), 3.45 (m, 0.5H, 4-H eq. Z isomer), 4.08 (t, 2H, OCH2), 4.52
(bb, 1H, OH), 7.99 (bb, 3H, NH3+); white solid: mp 152-155
°C. Anal. (C21H34N2O3‚HCl‚H2O) C, H, N, Cl, H2O.
(E,Z)-3-[2-(N,N-Dim et h yla m in o)et h oxyim in o]a n d r os-
ta n e-6r,17â-d iol (22h ). Prepared in 80% yield from 1c and
2-(N,N-dimethylamino)ethoxyamine dihydrochloride. The crude
product was purified by flash chromatography (CHCl3/MeOH/
26% NH4OH 90/10/1) to give 22h . 1H NMR (CD3OD) δ 0.75 (s,
3H, CH3), 0.94 (s, 3H, CH3), 2.30 (s, 6H, N(CH3)2), 2.64 (m,
2H, NCH2), 3.58 (m, 1H, 17-H), 4.11 (m, 2H, OCH2); white
solid: mp 102-106 °C. Anal. (C23H40N2O3‚H2O) C, H, N, H2O.
(E,Z)-3-(2-Am in oeth oxyim in o)a n d r osta n e-6r,17â-d iol
oxa la te (22i). Prepared in 85% yield from 1c and 2-amino-
ethoxyamine dihydrochloride. The crude product was purified
by flash chromatography (CHCl3/MeOH/26% NH4OH 90/10/
1). To the concentrated fractions a stoichiometric amount of
oxalic acid in MeOH was added. After addition of a 1/1 mixture
of EtOAc/Et2O, the precipitate was filtered to give 22i. 1H NMR
(DMSO-d6) δ 0.61 (s, 3H, CH3), 0.83 (s, 1.5H, CH3), 0.84 (s,
1.5H, CH3), 3.04 (t, 2H, NCH2), 3.07 (m, 0.5H, 2-H eq. E
isomer), 3.20 (m, 1H, 6-H), 3.42 (m, 1.5H, 4-H eq. Z isomer
and 17-H), 4.06 (t, 2H, OCH2), 7.71 (bb, 3H, NH3+); white
solid: mp 151-156 °C. Anal. (C21H36N2O3‚C2H2O4‚H2O) C, H,
N, H2O.
(Z)-3-(2-Tr iflu or oa ceta m id oeth oxyim in o)a n d r osta n e-
6r,17â-d iol (23a ) and (E)-3-(2-tr iflu or oa ceta m id oeth oxy-
im in o)a n d r osta n e-6r,17â-d iol (23b). To a suspension of
(E,Z) 3-(2-aminoethoxyimino)androstane-6R,17â-diol hydro-
chloride (22i) (1.10 g, 2.77 mmol) in THF (40 mL) at 0 °C,
(CF3CO)2O (0.86 mL, 6.18 mmol) was added dropwise. After
stirring at room temperature overnight, the mixture was
diluted with EtOAc (100 mL) and washed with a 5% aqueous
NaHCO3 solution (2 × 120 mL), H2O (100 mL), dried over Na2-
SO4, filtered, and evaporated to dryness. To the crude product,
MeOH (150 mL) and 5% aqueous NaHCO3 solution (40 mL)
were added. After stirring at 35 °C overnight, MeOH was
evaporated and the residue was extracted with EtOAc (2 ×
100 mL). The combined organic extracts were washed with
H2O (2 × 100 mL), dried over Na2SO4, filtered, and evaporated
to dryness. The residue was purified by flash chromatography
(CH2Cl2/EtOAc/acetone 7/2/1) to give 23a as a white foam (0.28
g, 22%) and 23b as a white foam (0.50 g, 40%). 23a : 1H NMR
(CDCl3) δ 0.73 (s, 3H, CH3), 0.92 (s, 3H, CH3), 3.48 (m, 1H,
6-H), 3.55 (m, 1H, 4-H eq.), 3.61 (t, 2H, NCH2), 3.64 (m, 1H,
17-H), 4.14 (m, 2H, OCH2), 7.40 (bb, 1H, CONH). 23b: 1H NMR
(CDCl3) δ 0.73 (s, 3H, CH3), 0.92 (s, 3H, CH3), 3.12 (m, 1H,
2-H eq), 3.48 (m, 1H, 6-H), 3.61 (m, 2H, NCH2), 3.64 (m, 1H,
17-H), 4.14 (m, 2H, OCH2), 7.30 (bb, 1H, CONH).
(E )-3-(2-Am in oe t h oxyim in o)a n d r ost a n e -6r,17â-d iol
h em ifu m a r a te (22j). To a solution of 23b (0.50 g, 1.08 mmol)
in MeOH (6.6 mL) and H2O (4.0 mL), K2CO3 (0.75 g, 5.42
mmol) was added. After stirring of the sample at room
temperature overnight, the solution was diluted with CH2Cl2
(50 mL) and 1 N NaOH (5 mL). The layers were separated
and the aqueous phase was extracted with CH2Cl2 (2 × 50 mL).
The combined organic extracts were dried over Na2SO4,
filtered, and evaporated to dryness. The residue was dissolved
in MeOH (4 mL) and fumaric acid (126 mg, 1.08 mmol) and
Et2O (2 mL) were added. The precipitate was filtered to give
22j (0.415 g, 80%). 1H NMR (DMSO-d6) δ 0.61 (s, 3H, CH3),
0.83 (s, 3H, CH3), 3.08 (m, 3H, NCH2 and 2-H eq.), 3.21 (m,
1H, 6-H), 3.42 (m, 1H, 17-H), 4.08 (t, 2H, OCH2), 7.78 (bb, 3H,
NH3+); white solid: mp 110-180 °C. Anal. (C21H36N2O3‚0.5
C4H4O4‚1.5 H2O) C, H, N, H2O.
(Z)-3-(2-Am in oe t h oxyim in o)a n d r ost a n e -6r,17â-d iol
h em ifu m a r a te (22k ). Prepared in 75% yield from 23a by the
procedure described above for the preparation of 22j. 1H NMR
(DMSO-d6) δ 0.61 (s, 3H, CH3), 0.84 (s, 3H, CH3), 2.89 (m, 2H,
NCH2), 3.21 (m, 1H, 6-H), 3.44 (m, 2H, 17-H and 4-H eq), 3.99
(t, 2H, OCH2); white solid: mp 180-183 °C. Anal. (C21H36N2O3‚
0.5 C4H4O4‚H2O) C, H, N, H2O.
(Z)-3-(3-Tr iflu or oacetam idopr opoxyim in o)an dr ostan e-
6r,17â-d iol (23c) and (E)-3-(3-tr iflu or oa ceta m id op r op oxy-
im in o)a n d r osta n e-6r,17â-d iol (23d ). Prepared from 1c and
3-aminopropoxyamine dihydrochloride by the general proce-
dure described above to give (E,Z)-3-(3-aminopropoxyimino)-
androstane-6R,17â-diol. Compounds 23c and 23d were pre-
pared from (E,Z)-3-(3-aminopropoxyimino)androstane-6R,17â-
diol by the general procedure described above for the preparation
of 23a and 23b. 23c (0.40 g, 25%), a white foam: 1H NMR
(MeOD) δ 0.74 (s, 3H, CH3), 0.98 (s, 3H, CH3), 3.35 (t, 2H,
NCH2), 3.37 (m, 1H, 6-H), 3.57 (m, 1 H, 4-H eq.), 3.58 (m, 1H,
17-H), 4.02 (m, 2H, OCH2). 23d (0.66 g, 41%), a white foam:
1H NMR (MeOD) δ 0.74 (s, 3H, CH3), 0.98 (s, 3H, CH3), 3.18
(m, 1H, 2-H eq), 3.35 (t, 2H, NCH2), 3.37 (m, 1H, 6-H), 3.58
(m, 1H, 17-H), 4.02 (m, 2H, OCH2).
(E)-3-(2-Am in op r op oxyim in o)a n d r ost a n e-6r,17â-d iol
h em ifu m a r a te (22l). Prepared in 82% yield from 23d by the
procedure described above for the preparation of 22j. 1H NMR
(DMSO-d6) δ 0.61 (s, 3H, CH3), 0.84 (s, 3H, CH3), 2.75 (m, 2H,
NCH2), 2.98 (m, 1H, 2-H eq), 3.21 (m, 1H, 6-H), 3.41 (m, 1H,
17-H), 3.98 (m, 2H, OCH2), 7.78 (bb, 3H, NH3+); white solid:
mp 147-181 °C. Anal. (C22H38N2O3‚0.5 C4H4O4‚0.5 H2O) C, H,
N, H2O.
(Z)-3-(2-Am in op r op oxyim in o)a n d r ost a n e-6r,17â-d iol
h em ifu m a r a te (22m ). Prepared in 80% yield from 23c by the
procedure described above for the preparation of 22j. 1H NMR
(DMSO-d6) δ 0.61 (s, 3H, CH3), 0.84 (s, 3H, CH3), 2.75 (m, 2H,
NCH2), 3.21 (m, 1H, 6-H), 3.40 (m, 2H, 17-H and 4-H eq), 3.98
(m, 2H, OCH2), 7.80 (bb, 3H, NH3+); white solid: mp 125-
174 °C. Anal. (C22H38N2O3‚0.5 C4H4O4‚H2O) C, H, N, H2O.
(E,Z)-3-(2-Am in oeth oxyim in o)-6â-h yd r oxya n d r osta n -
17-on e h yd r och lor id e (22n ). Prepared in 86% yield from 1f
and 2-aminoethoxyamine dihydrochloride. The product was
crystallized from MeOH/EtOAc. 1H NMR (DMSO-d6) δ 0.80
(s, 3H, CH3), 1.02 (s, 1.5H, CH3), 1.04 (s, 1.5H, CH3), 2.82 (dd,
0.5H, 4-H eq. Z isomer), 3.03 (t, 2H, NCH2), 3.06 (m, 0.5H,
2-H eq. E isomer), 3.65 (m, 1H, 6-H), 4.07 (t, 2H, OCH2), 4.45
(bb, 1H, OH), 7.95 (bb, 3H, NH3+); white solid: mp 160-170
°C dec. Anal. (C21H34N2O3‚HCl‚H2O) C, H, N, H2O.