Expedient synthesis of some novel pregnane derivatives and their evaluation as anti-oxidant and anti-dyslipidemic agents

Article abstract of DOI:10.1007/s00044-009-9280-y

Synthesis of a number of novel pregnane derivatives has been described in detail. They were prepared by reaction of 3β-hydroxy-5, 16-pregnadiene-20-one (2) with diethylene glycol, 2-methoxy ethanol, 1,2-propane diol, p-chloroaniline and p-fluoroaniline, respectively. The structures of these newly synthesized compounds have been established on basis of physical, analytical and spectral data. These pregnane derivatives have been evaluated for their anti-dyslipidemic activity (Triton model) and in vitro antioxidant activities. Out of the 12 compounds tested, 3 of them showed potent anti-dyslipidemic activity and 7 showed potent antioxidant and scavenger of oxygen free radicals. Springer Science+Business Media, LLC 2010.

Full text of DOI:10.1007/s00044-009-9280-y

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res (2011) 20:36–46
DOI 10.1007/s00044-009-9280-y
ORIGINAL RESEARCH
Expedient synthesis of some novel pregnane derivatives and their
evaluation as anti-oxidant and anti-dyslipidemic agents
•
Arun Sethi Gitika Bhatia Ashok K. Khanna
•
•
•
Mohammad Mobin Khan Abha Bishnoi
•
•
Anil K. Pandey Atul Maurya
Received: 2 April 2009 / Accepted: 3 December 2009 / Published online: 24 December 2009
ꢀ Springer Science+Business Media, LLC 2010
Abstract Synthesis of a number of novel pregnane
derivatives has been described in detail. They were pre-
pared by reaction of 3b-hydroxy-5, 16-pregnadiene-20-one
(2) with diethylene glycol, 2-methoxy ethanol, 1,2-propane
diol, p-chloroaniline and p-fluoroaniline, respectively. The
structures of these newly synthesized compounds have
been established on basis of physical, analytical and
spectral data. These pregnane derivatives have been eval-
uated for their anti-dyslipidemic activity (Triton model)
and in vitro antioxidant activities. Out of the 12 compounds
tested, 3 of them showed potent anti-dyslipidemic activity
and 7 showed potent antioxidant and scavenger of oxygen
free radicals.
suffering from diabetes accompanied with dyslipidemia,
have higher concentration of cholesterol, especially in low
density lipoprotein (LDL) and triglycerides, along with free
radical oxidative stress, have been recognized as the major
cause for the development of atherosclerosis, cardiovas-
cular diseases and coronary heart diseases (Alexander
et al., 2002). Hydroxyl free radicals (OH•) have also been
found to be responsible for the peroxidative damage to
lipoproteins present in the blood, which in turn are
responsible for the initiation and progression of athero-
sclerosis (Parthasarathy et al., 1992).
Therefore, to treat oxidative stress and disorders of lipid
metabolism, both together, may be a novel approach to
regress atherosclerosis and other cardiovascular diseases.
We have reported earlier that some precursors in hormone
synthesis, namely guggulsterone 4,17(20)-pregnadiene-3,
16 dione posses potent lipid lowering activity, together
with mild antioxidant effects and also inhibit oxidative
modification of LDL (Chander et al., 1996, Wang et al.,
2004). Recently, we found some pregnane derivatives
exhibited antioxidant and anti-dyslipidemic activities
altogether (Sethi et al., 2007, 2008). In continuation for
search and development of potent lipid lowering and
antioxidant drug, some new pregnane derivatives were
synthesized and tested in vivo and in vitro model for both
biological acitivities.
Keywords Steroids ꢀ Pregnanes ꢀ Oximo ether ꢀ
Anti-dyslipidemic activity ꢀ Antioxidant activity ꢀ
Cholesterol
Introduction
Dyslipidemia, a complication that is known to be mani-
fested with almost all the metabolic diseases in general and
diabetes in particular (Ruotola and Howard, 2002), is due
to abnormalities with the lipoprotein metabolism. It is well
established that dyslipidemia plays an important role in a
number of secondary complications in diabetes. Patients
Materials and methods
A. Sethi (&) ꢀ A. Bishnoi ꢀ A. K. Pandey ꢀ A. Maurya
Department of Chemistry, University of Lucknow, Lucknow
226007, India
Chemistry
e-mail: alkaarunsethi@rediffmail.com
All solvents used were of laboratory grade and were
purified and dried according to standard procedures. Col-
umn chromatography was performed with silica gel
G. Bhatia ꢀ A. K. Khanna ꢀ M. M. Khan
Division of Biochemistry, Central Drug Research Institute,
Lucknow 226001, India
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Med Chem Res (2011) 20:36–46
37
(60–120 mesh). IR spectra were recorded on Beckmann
Aculab-10, Perkin Elmer 881 and FTIR 8210 PC, Schi-
madzu spectrophotometers using KBr discs. Nuclear
magnetic resonance (NMR) spectra were recorded on
either Bruker advance DRX-300 MHz or Bruker DPX 200
FT spectrometers using TMS as an internal reference,
while 1H–1H–COSY correlations were recorded on
600 MHz Varian Inova Spectrometer. FAB mass spectra
were recorded on JEOL SX 102/DA 6000, using m-nitro-
benzyl alcohol as matrix (the matrix peak appeared at m/
z 136, 137, 154, 289 and 307), whereas ESI was recorded
on MICROMASS QUATTRO II triple quadrupole mass
spectrometer. Optical rotations were recorded on ORIBA,
SEPA-300 digital polarimeter. Chemical analysis was
carried out on Carlo-Erba-1108 instrument. The melting
points are recorded on an electrically heated melting point
apparatus and are uncorrected.
=NOCH, J = 6.4 Hz), 3.53 (m, 1H, H-3), 1.91(s, 3H, CH3-
21), 1.24 (d, 6H, =NOCH (CH3)2, J = 6.4 Hz), 1.038 (s,
3H, 19-CH3), 0.95(s, 3H, 18-CH3); 13C NMR (100 MHz,
CDCl3), d 152.32 (C-20), 151.31 (C-17), 141.24 (C-16),
130.97 (C-5), 121.41 (C-6), 74.93 (C-3), 71.82 (=NO–CH),
57.11 (C-14), 50.58 (C-9), 46.75 (C-13), 42.36 (C-4), 37.57
(C-10), 37.18 (C-12), 36.71 (C-1), 35.85 (C-15), 31.63
(C-8), 30.38 (C-7), 29.68 (C-2), 21.82 (=NCH(CH3)2,
21.03 (C-11), 19.35 (C-19), 15.97 (C-21), 11.77 (C-18);
ESI MS 372(M??1), 354 (M?-H2O), 312; 280. Anal.
calcd. for C24H37NO2: C: 77.58, H: 10.04, N: 3.77%.
Found: C: 7.32, H: 9.86, N: 3.79%.
General procedure for the synthesis of compounds 5, 6
and 7
550 mg (1.75 mmol) of compound 2 was dissolved in
4.2 ml (35 mmol) of freshly distilled diethylene glycol and
then 0.5 ml of boron trifluoride etherate was added to it.
The reaction mixture was stirred at 30ꢁC for 55–60 h
(reaction was monitored by t.l.c during this period). Ice-
cold solution of sodium bicarbonate was added to light
brown solution of reaction mixture and the aqueous phase
was extracted with dichloromethane. The organic layer was
washed with water, dried over anhydrous sodium sulphate
and then concentrated under reduced pressure. The crude
product was purified by column chromatography on silica
gel (hexane/ethyl acetate 80:20) to afford the pure
compound.
3b-Acetoxy-5, 16-pregnadiene-20-one (1)
Diosgenin was converted into compound 1 by reported
method (Marker, 1949; Muller and Norton, 1955). It was
identified (Fukushima and Gallagher, 1951) by its m.p.
1
166–170ꢁC, H NMR and MS.
3b-Hydroxy-5, 16-pregnadiene-20-one (2)
Deacetylation of compound 1 by Zemplen method (Stubbs
et al., 2006) yielded compound 2, identified by its m.p.
212–214ꢁC (lit m.p. 216ꢁC) (Kaneko et al., 1973; Sethi
1
et al., 2007) H NMR and MS.
3b-Hydroxy-16a-[2(2-hydroxy ethoxy)-ethoxy]pregn-5-en-
20-one (5)
3b-Acetoxy-5, 16-pregnadiene-20-one oxime (3)
Yield 80%, [a]D -81.8ꢁ (c, 0.55, CHCl3); 1H NMR
(CDCl3, 200 MHz), d (ppm) 5.35 (1H, m, H-6); 4.52 (1H,
m, H-16), 3.52–3.74 (9H, m, OCH2CH2OCH2CH2 and H-
3), 2.62 (1H, d, H-17, J = 6.4 Hz); 2.17 (3H, s, CH3-21);
1.00 (3H, s, CH3-19), 0.64 (3H, s, CH3-18); 13C NMR
(75 MHz,CDCl3), d 208.3 (C-20), 140.7 (C-5), 121.2
(C-6), 80.15 (C-16), 72.54 (C-3), 72.27 (OCH2CH2-
OCH2CH2), 71.47 (OCH2CH2O), 70.36 (OCH2CH2O),
68.98 (OCH2CH2OH), 61.68 (C-17), 54.43 (C-14), 49.80
(C-9), 44.41 (C-13), 42.09 (C-4), 38.68 (C-12), 37.11
(C-1), 36.44 (C-10), 32.23 (C-15), 31.74 (C-8), 31.43 (C-7
and C-2), 29.63 (C-21), 20.69 (C-11), 19.33 (C-19), 14.38
(C-18); FAB MS (m/z) 421 (M??1), 391, 374, 362, 285,
271, 267, 224, 163, 133, 121, 109, 105.
Compound 1 was converted into compound 3 by reported
method (Rosenkranz et al., 1956). It was identified by its
1
m.p. 225–228ꢁC, H NMR and MS.
20-(O-isopropyl)-oximino-3b-hydroxy-pregn-5,
16-diene (4)
Compound
3
(500 mg, 1.40 mmol), NaH (96 mg,
4.01 mmol) in dry DMF (25 ml) was stirred at 0ꢁC for
30 min. To the reaction mixture, 2-bromopropane (0.5 ml)
was added and the reaction mixture was further stirred at
room temperature for 3 h. DMF was evaporated under
reduced pressure and the solid residue obtained was
extracted with chloroform, washed with water, dried over
anhydrous sodium sulphate and concentrated in vaccuo.
Column chromatography of the resultant residue afforded
compound 4 as white solid. Yield 65%, m.p 115–118ꢁC,
[a]D -18ꢁ (c 0.08, CHCl3); 1H NMR (400 MHz, CDCl3), d
ppm 5.96 (m, 1H, H-16), 5.36 (m, 1H, H-6), 4.29 (q, 1H,
3b-Hydroxy-16a-(2-methoxyethoxy) pregn-5-en-20-one (6)
Yield 70%, m.p 155–157ꢁC, [a]D ?64.28 (c, 0.30, CHCl3);
1H NMR (CDCl3, 200 MHz), d (ppm) 5.35 (1H, m, H-6);
4.47 (1H, m, H-16), 3.53–3.39 (5H, m, OCH2CH2 and H-
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Med Chem Res (2011) 20:36–46
3), 3.35(3H, s, OCH3), 2.65 (1H, d, H-17, J = 6.4 Hz);
2.17 (3H, s, CH3-21); 1.17 (3H, s, CH3-19), 0.99 (3H, s,
CH3-18); FAB MS (m/z) 391(M??1), 347, 271, 253, 163,
145, 138, 109, 105. Anal. calcd. for C24H38O4: C: 73.81, H:
9.81%. Found: C: 73.68, H: 9.72%.
(3H, s, CH3-19), 0.62 (3H, s, CH3-18); ESI MS m/z 432
(M?), 434 (M??2), 389.
3b-Acetoxy-16a-(2-acetoxy propoxy)-pregn-5-en-
20-one (10)
Yield, 80%, [a]D -18ꢁ (c,CHCl3); 1H NMR (CDCl3,
400 MHz), d (ppm) 5.38 (1H, m, H-6); 5.01–4.93 (1H, m,
OCH2CHOCOCH3CH3), 4.64–4.56 (1H, m, H-3),
4.46–4.43 (1H, m, H-16), 3.42–3.36 (1H, m, OCH2),
3.32–3.26 (1H, m, OCH2), 2.58 (1H, d, H-17, J = 6.0 Hz);
2.16 (3H, s, CH3-21); 2.03 (6H,s, OAc), 1.21 (3H, d,
OCH2CHOAcCH3, J = 8.0 Hz), 1.01 (3H, s, CH3-19),
0.62 (3H, s, CH3-18); ESI MS m/z 475 (M??1), 415,387,
371, 355, 297, 253.
3b-Hydroxy-16a-(2-hydroxy propoxy) pregn-5-en-
20-one (7)
Yield 75%, m.p 122–125ꢁC [a]D -40ꢁ (c, 0.06, CHCl3); 1H
NMR (CDCl3, 300 MHz), d (ppm) 5.38 (1H, m, H-6); 4.51
(1H, m, H-16), 3.95 (1H, m, OCH2CHOHCH3), 3.55 (1H,
m, H-3) 3.41 (1H, dd, OCHaHb, J = 6 and 3 Hz), 3.21 (1H,
dd, OCHaHb, J = 6 and 3 Hz), 2.60 (1H, d, H-17,
J = 6.0 Hz); 2.20 (3H, s, CH3-21); 1.16 (3H, d,
OCH2CHOHCH3, J = 6.0 Hz), 1.02 (3H, s, CH3-19), 0.65
(3H, s, CH3-18); 13C NMR (75 MHz, CDCl3), d 210.10
(C-20), 140.75 (C-5), 121.19 (C-6), 80.29 (C-16), 75.02
(C-10), 71.62 (C-3), 66.79(C-20), 66.22 (C-17), 54.49
(C-14), 49.83 (C-9), 44.36 (C-13), 42.19 (C-4), 38.75
(C-12), 37.12 (C-1), 36.49 (C-10), 35.03 (C-15), 32.37 (C-
8), 31.55 (C-7), 31.46 (C-2), 29.88 (C-21), 20.72 (C-11),
19.36 (C-19), 18.55 (C-30) 14.41 (C-18); ESI MS (m/z) 390
(M?) 371, 360, 299, 271, 303; Anal. calcd. for C24H38O4:
C: 73.81, H: 9.81%. Found: C: 73.64, H: 9.74%.
General procedure for the synthesis of compounds 11
and 12
Compound 5 (435 mg) in dry tetrahydrofuran (7.1 ml) and
dry methanol (2.1 ml) was treated with sodium borohy-
dride (84 mg, 2.21 mmol) portionwise during 10 min at
5ꢁC; reaction mixture was stirred at the same temperature
for 1 h, when t.l.c showed completion of reaction. Acetic
acid was added dropwise until the reaction mixture became
neutral. Water was added and the resulting mixture con-
centrated under reduced pressure. The aqueous phase was
extracted with dichloromethane. The organic layer was
washed with water, dried over anhydrous sodium sulphate
and then concentrated under reduced pressure. The crude
product was purified by column chromatography on silica
gel (hexane/ethyl acetate 75:25) to afford the pure
compound.
General procedure for the synthesis of compounds 8, 9
and 10
Conventional acetylation of compounds 5, 6 and 7 with
acetic anhydride and pyridine yielded 8, 9 and 10,
respectively, as syrupy compounds.
3b-Acetoxy-16a-[2(2-acetoxy ethoxy)-ethoxy]-pregn-5-en-
20-one (8)
3b, 20b-Dihydroxy-16a-[2(2-hydroxyethoxy)-ethoxy]-
pregn-5-en (11)
Yield 85%, [a]D -177.78ꢁ (c, 0.30, CHCl3); 1H NMR
(CDCl3, 200 MHz), d (ppm) 5.36 (1H, m, H-6); 4.65 (1H,
m, H-16), 4.25–4.08 (3H, m, OCH2CH2OCH2CH2 and H-
3), 3.72–3.64 (4H, m, OCH2CH2OCH2CH2), 3.59–3.51
(3H, m, OCH2CH2OCH2CH2), 2.62 (1H, d, H-17,
J = 6.0 Hz); 2.17 (3H, s, CH3-21); 2.08(3H,s, OAc),
2.02(3H,s, OAc), 1.01 (3H, s, CH3-19), 0.88 (3H, s, CH3-
18); ESI MS m/z 504 (M?), 444, 355, 313, 253.
Yield 75% m.p. 140–145ꢁC, [a]D -71.5ꢁ (c, 0.50, CHCl3);
1H NMR (CDCl3, 200 MHz), d (ppm) 5.36, (1H, m, H-6);
4.41(1H, m, H-16), 3.95–3.87 (1H, m, H-20), 3.71–3.49
(9H, m, OCH2CH2OCH2CH2OH and H-3), 1.25 (3H, d,
CH3-21, J = 6.0 Hz); 1.01 (3H, s, CH3-19), 0.80 (3H, s,
CH3-18); FAB MS (m/z) 421 (M?-1), 316, 271, 145, 138,
120, 105. Anal. calcd. for C25H42O5: C: 71.05, H: 10.02%.
Found: C: 70.94, H: 9.92%.
3b-Acetoxy-16a-(2-methoxy ethoxy)-pregn-5-en-20-one (9)
3b, 20b-Dihydroxy-16a-(2-methoxy ethoxy)-pregn-
5-en (12)
Yield 85%, [a]D ?56.50ꢁ (c, 0.25, CHCl3); 1H NMR
(CDCl3, 200 MHz), d (ppm) 5.35 (1H, m, H-6); 4.51 (1H,
m, H-3), 4.46 (1H, m, H-16), 3.49-3.46 (4H, m, OCH2-
CH2OCH3), 3.35(3H, s, OCH3), 2.60 (1H, d, H-17,
J = 6.0 Hz); 2.17 (3H, s, CH3-21); 2.05(3H, s, OAc), 1.01
Yield 75%, m.p. 185–187ꢁC, [a]D -138.7(c, 0.40, CHCl3);
1H NMR (CDCl3, 200 MHz), d (ppm) 5.33 (1H, m, H-6);
4.39–4.32 (1H, m, H-16), 3.90–3.80 (1H, m, H-20),
3.71–3.50 (5H, m, OCH2CH2OCH3 and H-3), 3.36 (3H, s,
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Med Chem Res (2011) 20:36–46
39
OCH3), 1.25 (3H, d, CH3-21, J = 6.0 Hz); 1.05 (3H, s,
CH3-19), 0.79 (3H, s, CH3-18); MS (m/z) 391 (M?-1),
331, 329, 313, 297, 271, 145, 105. Anal. calcd. for
C24H40O4: C: 73.43, H: 10.29%. Found: C: 73.24, H:
10.20%.
(hexane/ethyl acetate 80:20) to afford the pure compound
14. Yield: 80%, m.p. 144–146ꢁC; [a]D -38 (c, 0.1,
CHCl3); 1H NMR (CDCl3, 200 MHz) d (ppm) 6.86 (2H, m,
Ar–H)), 6.53 (1H, m, Ar–H), 5.34 (1H, m, H-6); 4.39 (1H,
t, H-16, J = 8.0 Hz), 3.52(1H, m, H-3), 2.41 (1H, d, H-17,
J = 7.4 Hz); 2.15 (3H, s, CH3-21); 1.01 (3H, s, CH3-19),
0.72 (3H, s, CH3-18); 13C NMR (100 MHz, CDCl3, d):
207.68 (C-20), 157.16 (ArC-50), 154.1 (C-10), 143.80 (C-5),
140.75 (C-20), 12179 (C-6), 115.87 (C-30), 115.68 (C-60),
114.77(C-40), 72.71 (C-3), 71.57 (C-17), 55.08 (C-14),
53.85 (C-9), 50.31 (C-16), 4450 (C-13), 42.48 (C-4), 38.95
(C-12), 37.14(C-1), 36.49 (C-10), 33.66 (C-15), 32.22
(C-8), 31.61 (C-7), 31.50 (C-2), 29.31 (C-21), 20.78
(C-11), 19.33 (C-19), 14.22 (C-18); FAB MS (m/z)
425(M?),426(M??1), 383, 257, 239, 138, 105. Anal.
calcd. for C27H36FNO2: C: 76.20, H: 8.53, N: 3.29%
Found: C: 75.78, H: 8.26, N: 3.21%.
3b-Hydroxy-16a-(4-chloro phenylamino)-pregn-5-en-20-
one (13)
550 mg (1.75 mmol) of compound 2 was dissolved in
50 ml of freshly distilled dry methanol and then 200 mg of
p-chloroaniline was added to it. The reaction mixture was
refluxed for 20 h (reaction was monitored by t.l.c during
this period). Ice-cold solution of sodium bicarbonate
was added to the reaction mixture and the aqueous phase
was extracted with dichloromethane. The organic layer was
washed with water, dried over anhydrous sodium sulphate
and then concentrated under reduced pressure. The crude
product was purified by column chromatography on silica
gel (hexane/ethyl acetate 80:20) to afford the pure com-
pound 13 as white solid. Yield: 80%, m.p. 205–207ꢁC; [a]D
-36ꢁ(c, 0.40, CHCl3); IR (KBr) mmax 3390, 3247, 1685,
3b-Hydroxy-16a, 17a-epoxypregn-5-en-20-one (15)
Compound 1 was converted into compound 15 by reported
method (Das and Kirk, 1984; Neher et al., 1958). It was
identified by its m.p. 212–214ꢁC (lit m.p. 216ꢁC), 1H NMR
and MS.
1
1596, 815 cm-1; H NMR (CDCl3, 200 MHz), d (ppm)
7.08 (2H, d, Ar–H-30 and H-50, J = 8.8 Hz), 6.52 (2H, d,
Ar–H-20 and H-60, J-8.8 Hz), 5.33 (1H, m, H-6); 4.40 (1H,
t, H-16, J = 8.0 Hz), 3.67 (1H, m, N–H), 3.52(1H, m, H-
3), 2.45 (1H, d, H-17, J = 7.4 Hz); 2.15 (3H, s, CH3-21);
1.01 (3H, s, CH3-19), 0.71 (3H, s, CH3-18); 13C NMR
(50 MHz, CDCl3, d): 207.94 (C-20), 146.45 (ArC-10),
141.18 (C-5), 129.42 (C-30 and C-50), 122.61 (C-6), 121.52
(C-40), 115.27 (C-20 and C-60), 73.15 (C-3), 72.01 (C-17),
55.55 (C-14), 53.62 (C-9), 50.31 (C-16), 44.98 (C-13),
42.59 (C-4), 39.39 (C-12), 37.58 (C-1), 36.92 (C-10), 34.15
(C-15), 32.32 (C-8), 32.04 (C-7), 31.94 (C-2), 30.09
(C-21), 21.22 (C-11), 19.79 (C-19), 14.67 (C-18); FAB MS
(m/z) 441(M?, Cl35),443(M?, Cl37) 426, 391, 388, 253,
137, 133, 105; Anal. calcd. for C27H36ClNO2: C: 73.36, H:
8.21, N: 3.17% Found: C: 73.02, H: 8.04, N: 3.08%.
3b, 17a-Dihydroxy-16a-(2-hydroxy ethoxy) pregn-5-en-20-
one (16)
580 mg (1.75 mmol) of compound 15 was dissolved in
4.2 ml (35 mmol) of freshly distilled ethylene glycol. The
reaction mixture was stirred at 30ꢁC for 25–30 h (reaction
was monitored by t.l.c during this period). Ice-cold solution
of sodium bicarbonate was added and the aqueous phase
was extracted with dichloromethane. The organic layer was
washed with water, dried over sodium sulphate and then
concentrated under reduced pressure. The crude product
was purified by column chromatography on silica gel
(hexane/ethyl acetate 80:20) to afford the pure compound.
Yield: 65%, m.p. 185–190ꢁC [a]D -33 (c, 0.12, CDCl3);
1H NMR (CDCl3, 200 MHz), d (ppm) 5.41 (1H, m, H-6);
4.13 (1H, m, H-16), 3.73–3.71 (2H, m, OCH2CH2OH),
3.54–3.41 (3H, m, OCH2CH2OH and H-3), 2.13 (3H, s,
CH3-21), 0.98 (3H, s, CH3-19), 0.84 (3H, s, CH3-18); 13C
NMR (75 MHz, CDCl3), d 208.6 (C-20), 141.9 (C-5),
121.57 (C-6), 92.3 (C-17), 81.2 (C-16), 75.13 (OCH2-
CH2OH), 71.9 (C-3), 65.05 (OCH2CH2OH), 59.03 (C-14),
48.58 (C-9), 44.2 (C-13), 42.12 (C-4), 40.04 (C-12), 37.34
(C-1), 36.92 (C-10), 36.89 (C-15), 32.86 (C-8), 32.19
(C-7), 31.43 (C-2), 29.68 (C-21), 20.51 (C-11), 18.67
(C-19), 12.10 (C-18); ESI MS m/z 392 (M?), 349, 331,
285, 164. Anal. calcd. for C23H36O5: C: 70.38, H: 9.24%
Found: C: 70.22, H: 9.13%.
3b-Hydroxy-16a-(2-amino-5-fluoro-phenyl)-pregn-5-en-
20-one (14)
550 mg (1.75 mmol) of compound 2 was dissolved in
50 ml of freshly distilled dry methanol and then 200 mg of
p-fluoroaniline and 0.5 ml of boron trifluoride etherate was
added to it. The reaction mixture was refluxed for 20 h
(reaction was monitored by t.l.c during this period). Ice-
cold solution of sodium bicarbonate was added to the
reaction mixture and the aqueous phase was extracted with
dichloromethane. The organic layer was washed with
water, dried over anhydrous sodium sulphate and then
concentrated under reduced pressure. The crude product
was purified by column chromatography on silica gel
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Biological evaluation
(of the side chain at C-16) and one methine proton at C-3,
together with a one proton multiplet at d 4.52 due to
methine proton at C-16 suggested the introduction of 2(2-
hydroxy ethoxy)-ethoxy group at C-16. The absence of
vinylic proton signal of C-16 and the presence of one
proton doublet at d 2.62 presumably due to C-17 proton
confirmed the introduction of this group at C-16. The
magnitude of the coupling of C-17 proton doublet
(J = 6.4 Hz) due to J16bH-17aH confirms the orientation of
the side chain at C-16 to be a. The FAB MS of 5 showed
protonated molecular ion peak at m/z 421. The retro Diels–
Alder fragmentation of d5 bond yielded the fragment at
m/z 224 and m/z 105 further conforming the introduction of
2(2-hydroxy-ethoxy)-ethoxy group at C-16. In the 13C
NMR spectrum presence of 5, carbon signals at d 80.15,
72.27, 71.47, 70.71 and 68.98 further substantiated the
proposed structure Similarly the structure of 6 was con-
firmed on the basis of spectral data. The IH NMR spectrum
showed the appearance of three proton singlet at d 3.35 for
methoxy group along with a five proton multiplet in the
region d 3.53–d 3.39, which are due to two methylene
groups and one methine proton at C-3, together with a one
proton doublet at d2.65 (J = 6.4 Hz) suggested the intro-
duction of 2-methoxy ethoxy group at C-16. The FAB MS
showed protonated molecular ion peak at m/z 391, besides
other important fragments at m/z 347 and 271 further
The procedures adopted to evaluate the anti-dyslipidemic;
anti-dyslipidemic activity in high-fat diet fed hamster
model and anti-oxidant activity of pregnane derivatives
was same at reported earlier (Sethi et al., 2007; Bhatia
et al., 2008). In addition, the post-heparin lipolytic activity
(PHLA) and protein were also assayed by adopting known
procedures (Wing and Robinson, 1968; Lowry et al.,
1951).
Animals used: Rats (Charles Foster strain, male, adult,
body weight 200–225 g) were kept in a room with con-
trolled temperature (25–26ꢁC), humidity (60–80%) and 12/
12 h light/dark cycle (light on from 8.00 a.m to 8.00 p.m)
under hygienic conditions. Animals, which were acclima-
tized for 1 week before starting the experiment, had free
access to the normal diet and water ad libitum. Post heparin
lipolytic activity was checked after oral feeding of preg-
nane derivatives in triton induced rats. In fasted rats after
18 h, 30 units heparin was injected intravenously to each
rat. After 10 min of interval, blood was withdrawn from
retro-orbital plexus and assayed the activity (Wing and
Robinson, 1968) and protein was also estimated in the
serum samples by the Lowry–Rosenbrough method (Lowry
et al., 1951).
1
confirmed the presence of side chain at C-16. In the H
Result and discussion
NMR spectrum of compound 7, each hydrogen of the
methylene appeared as doublet of doublet at d 3.21 and d
3.41 (J = 6 and 3 Hz). Besides this the methine proton
appeared as a multiplet at d 3.95 and the methyl appeared
as a doublet at d 1.16 (J = 6.0 Hz). In the 13C NMR
spectrum of 7, carbon signals at d 80.29, 75.02, 66.79 and
18.55 further substantiated the introduction of the side
chain at C-16.
Chemistry
Diosgenin was converted to compound 1 by earlier repor-
ted method (Marker, 1949; Muller and Norton, 1955).
Reaction of compound 1 with hydroxylamine hydrochlo-
ride in presence of sodium acetate yielded 3 (Rosenkranz
et al., 1956). A novel steroidal oximino ether derivative 4
was synthesized by treating 3 with 2-bromopropane in dry
DMF in presence of base NaH (Jammart-Gregore et al.,
1989). In the 1H NMR spectrum of 4, besides other peaks, a
one-proton quartet at d 4.29 (J = 6.4 Hz) along with a six-
proton doublet at d 1.24 (J = 6.4 Hz) confirmed the pro-
posed structure. In the 13C NMR spectrum of 4, presence of
carbon signals at d 71.82 (=NOCH) and d 21.82 [=NOCH
(CH3)2] further substantiated the proposed structure. The
ESI MS of 4 recorded the highest mass ion peak at m/z 372
corresponding to M??1.
Acetylation of 5, 6 and 7 yielded the expected diacetyl,
monoacetyl and diacetyl derivatives 8, 9 and 10, respec-
tively (Scheme 1), whose structures were confirmed on the
basis of spectral data. The 1HNMR of 8 was better resolved
(in comparison to parent compound, 5) and more helpful in
identification of the methylene protons of the side chain (at
C-16). A three-proton multiplet, which appeared downfield
from d 4.25 to d 4.08, may be attributed to the 40 methylene
protons and one methine proton at C-3. A four proton
multiplet from d 3.72 to d 3.64 and a two proton multiplet
from d 3.59 to d 3.51 may be due to methylene protons at
C-30, C-20 and C-10, respectively.
Synthesis of C-16-substituted pregnane derivatives was
carried out by treating compound 2 with different nucleo-
philic reagents like diethylene glycol, 2-methoxy ethanol
and 1,2 propane diol in the presence of BF3:Et2O as
catalyst.
In the next step of the synthesis, reduction of 5 and 6
with sodium borohydride afforded corresponding 11 and 12
as the major isolable products (Scheme 1). Structures of
both the compounds were confirmed on the basis of spec-
tral analysis. In the 1H NMR spectrum of 11, the absence of
signal around d 2.2 due to C-21 methyl and the appearance
1
In the H NMR of 5, appearance of broad multiplet in
the region d 3.52–d 3.74, which is due to methylene groups
123

Med Chem Res (2011) 20:36–46
41
Scheme 1 Synthesis of
compounds 1–16
of a three-proton doublet at d 1.25 (J = 6 Hz) due to C-21
secondary methyl group together with a one-proton mul-
tiplet from d 4.86 to d 4.01 due to carbinol methine proton
at C-20 confirmed the reduction of carbonyl group. The
FABMS did not record the M? or protonated molecular ion
peak. However, the fragments at m/z 215 and m/z 155
definitely proved the presence of hydroxy ethyl and [2(2
hydroxy ethoxy)-ethoxy] chains at C-17 and C-16,
respectively. The stereochemistry of the orientation of the
hydroxyl group at C-20 in 11 and 12 is postulated to be b as
suggested by the fact that C-20 carbonyl groups on
reduction with metal hydrides give 80% 20b alcohols
(Lewbart, 1968). Besides this the presence of small NOE
between C-13 methyl and H-20 proton confirmed the
hydroxyl group at C-20 to be b (R) (Kirk, 1993).
and a one-proton doublet at d 2.45 (7.4 Hz) suggested the
introduction of p-chloroaniline group at C-16. In the 13C
NMR spectrum of 13, carbon signal at d 146.45, 129.42,
121.52, 115.27 and 50.31 further substantiates the proposed
structure. The FAB MS of 13 recorded the molecular ion
peak at m/z 441 (Cl35) (100%) and m/z 443 (Cl37). The
1
proposed structure was further confirmed by the H–1H–
COSY correlation experiment. Compound 2 when refluxed
with p-fluoroaniline in presence of Lewis acid catalyst
(BF3ꢀEt2O) yielded compound 14 (Scheme 1). The pres-
ence of two-proton multiplet at d 6.86, one proton multiplet
at d 6.53 along with one proton triplet at d 4.39
1
(J = 6.8 Hz) and one proton doublet at d 2.45 in its H
NMR spectrum confirmed the proposed structure.
Reaction of epoxide derivative 15 with ethylene glycol
1
led to the formation of compound 16 (Scheme 1). The H
Compound 2 was refluxed with p-chloroaniline (in
absence as well as in presence of boron trifluoride etherate)
yielding compound 13. In the 1H NMR spectrum of 13, two
doublets of two protons each at d 7.08 (J = 8.8 Hz) and d
6.52 (J = 8.8 Hz), one-proton triplet at d 4.40 (J = 8 Hz)
NMR spectrum of 16 showed one proton multiplet at d 4.13
for H-16, two-proton multiplet from d 3.73 to d 3.71
(OCH2CH2OH) and another three-proton multiplet from d
3.54 to d 3.41 (OCH2CH2OH and H-3) suggested the
123

42
Med Chem Res (2011) 20:36–46
introduction of this group at C-16. The ESI MS of 16
recorded the molecular ion peak at m/z 392.
Biological results
The present study has been undertaken to evaluate the anti-
dyslipidemic and anti-oxidant activity of pregnane deriv-
atives. Administration of triton WR-1339 caused a marked
increase in the serum levels of TC (3.21F), PL (3.97F), TG
(3.39F), PHLA (36%) and protein (1.8F), respectively
(Table 1). A significant reversal was noticed in the treat-
ment of hyperlipidemic rats with pregnane derivatives at
the dose of 400 mg/kg p.o. reversed the plasma levels of
lipid with varying extents. The effect of compounds 7, 8,
13 and 14 caused decrease in plasma levels of lipids TC,
PL, TG, and protein by 20, 24, 23, 21%, 18, 23, 25, 24%,
18, 23, 25, 23% and 22, 24, 24, 21%, respectively. While
compounds 4, 5, 6, 9, 10, 11, 12 and 16 showed mild lipid
lowering activity as compared to triton, which was more
significant than other compounds. These data were com-
pared with standard drug Gemfibrozil at the dose of
100 mg/Kg, showed decrease in plasma levels of TC, PL,
TG and protein by 36, 34, 36 and 30%, respectively. The
post heparin lipolytic activity was partially activated in
plasma of hyperlipidemic rats. The post heparin lipolytic
activity had a significant inhibition by 23, 20, 21, 23, 31
and 24% (Table 1). However, Gemfibrozil causes signifi-
cant reversal of these enzymes levels (Table 1). Triton
WR-1339 acts as a surfactant, suppresses the action of
lipases and blocks the uptake of lipoproteins from the
circulations of extrahepatic tissues resulting in increase in
the level of circulatory lipids (Schurr et al., 1972). Fur-
thermore, Triton WR-1339 is known to cause structural
modification in circulatory lipoprotein, so that they fail to
interact with capillary lipoprotein lipases. Above test
compound may have interfere with clustering lipoprotein
with Triton. In this way, lipoprotein may freely catabolize
by these enzymes.
Compounds 8, 13 and 14 were also tested in high fat diet
(HFD) model. In chronic experiments, feeding with high
fat diet once a day for 30 days produced hyperlipidemia.
These compounds were fed orally (100 mg/kg, b.w)
simultaneously with HFD. Control animals received the
same amount of normal saline or groundnut oil. At the end
of the experiment, rats were fasted overnight and blood was
withdrawn. The animals were killed and their liver was
promptly excised. Serum was analyzed for their TC, PL,
TG and plasma lecithin. Cholesterol acyltransferase activ-
ity (LCAT) and PHLA activity were assayed. In hyper-
lipidemic rats, hyperlipidemia was evidenced by increase
in the plasma level of TC (2.55F), PL (1.93F), TG (2.00F)
and inhibition of hepatic lipolytic activity (52%).
123

Med Chem Res (2011) 20:36–46
43
Treatment with compounds 8, 13 and 14 significantly
restored the enzyme activity 15, 18 and 27%, respectively,
and standard drug Gemfibrozil (100 mg/kg) showed 33%
activity. Compound 8, 13, 14 and Gemfibrozil showed
potent lipid lowering activity by TC (21, 19, 22 and 28%),
PL (29, 30, 28 and 32%), TG (23, 23, 20, 31%), respec-
tively (Table 2). In HFD rat’s hyperlipidemia as evidenced
by increased hepatic lipids TC (1.56F), PL (1.52F), TG
(1.59F) and compounds 8, 13 and 14 and gemfibrozil
showed potent hepatic lipid lowering activity by TC (21,
19, 22 and 28%), PL (29, 30, 28, 32%), TG (23, 23, 20 and
31%), respectively (Table 3).
The stimulation of plasma LCAT and hepatic lipases is
the mechanisms responsible for a significant lowering of
b-lipoproteins–lipid. Compound 8, 13 and 14 caused sig-
nificant increase in plasma levels of lipid in triton as well
as HFD model of hyperlipidemia. Though the lipid low-
ering activity of these compounds was comparatively less
than Gemfibrozil, however, we are in process to synthesize
more potent pregnane derivatives.
Hyperlipidemia may also induce other abnormalities
like oxidation of fatty acids, leading to the formation of
ketone bodies as well as marked liver and muscle resis-
tance to insulin, which initiates the progression of diabetes
in patients (Stehouwer et al., 1997). The effect of these
compounds on triglycerides lowering was also observed
through reversal of post heparin treated animals. There is a
significant correlation between the ability of tissue to
incorporate free fatty acid by hydrolysis of lipoprotein
triacylglycerol and the enzyme lipoprotein lipase. These
test samples inhibited cholesterol biosynthesis and poten-
tiated the activity of lipolytic enzymes to early clearance of
lipids from circulation in triton-induced hyperlipidemia.
We have successfully used this model for evaluation of
lipid lowering activity of pregnane derivatives. Data of
Table 1 suggest the restoration of PHLA activity on
treatment with pregnane derivatives. Furthermore due to
hyperglycemia, increase in non-enzymic glycosylation
occurs, accompanied with glucose oxidation and these
reactions being catalyzed by Cu?2 and Fe?2, resulting in
the formation of O•2 and OH• radicals, which further
accelerate the risk of cardiac diseases in dyslipidemic
patients (Asahina et al., 1995). This compound may
enhance the synthesis of LDL-apoprotein (apo B) as well
as receptor protein and inhibit the oxidative modification of
LDL to accelerate the turnover of LDL-cholesterol in liver
(Windler et al., 1980).
Effects of compounds 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 and
16 on oxygen free radical generation in vitro: the scavenging
potential of pregnane compounds at 100 and 200 lg/ml
against formation of O-2 and OH• in non-enzymic system
were also studied (Table 4). The significant decrease in
superoxide anions by (20–34%), hydroxyl radicals
123

44
Med Chem Res (2011) 20:36–46
Table 3 Effect of pregnane derivatives on liver biochemical parameters and faecal bile acid excretion in hyperlipidemic rats
Parameters
LPL activity
Total cholesterola (TC)
Triglyceridea (TG)
Phospholipidsa (PL)
Control
135.77 10.81
6.37 0.14
9.33 0.23
24.14 1.73
Cholesterol treated
Cholesterol ? 8
Cholesterol ? 13
Cholesterol ? 14
65.66 4.82 (-52)*** 10.00 0.32 (?1.56F)*** 14.88 1.00 (?1.59F)*** 36.82 2.12 (-1.52F)***
77.16 15.21 (?15)*
80.58 6.14 (?18)**
90.31 7.89 (?27)***
7.89 0.13 (-21)**
8.12 0.12 (-19)***
7.84 0.11 (-22)**
11.38 0.28 (-23)***
11.50 0.19 (-23)***
11.84 0.37 (-20)**
10.26 0.74 (-31)***
26.14 176 (-29)***
25.72 1.64 (-30)***
26.18 1.37 (-28)***
24.89 1.68 (-32)***
Cholesterol ? gemfibrozil
standard drug
98.92 82.14 (?33)*** 7.12 0.10 (-28)***
Unit: a n.mol free fatty acid formed h/mg protein. Values are mean SD of six animals. * P \ 0.05, ** P \ 0.01, *** P \ 0.001. Cholesterol
fed hyperlipidemic groups was compared with control. Cholesterol plus drug and gemfibrozil treated groups with cholesterol
Table 4 Effect of pregnane derivatives on free radical generation and lipid peroxidation in rat liver microsomes invitro
Treatment
Conc. (lg/ml)
Formation of superoxide anionsa
Formation of hydroxyl radicalsb
Microsomal lipid peroxidationc
4
Control 75.64 5.11
Control 70.12 5.32
Control 85.55 6.33
100
200
Exp 68.92 4.83 (-9) NS
Exp 60.12 5.00 (-20)**
Control 82.34 7.12
Exp 58.44 4.11 (-17)**
Exp 50.81 3.81 (-27)***
Control 72.11 5.61
Exp 64.44 5.11 (-25)***
Exp 56.66 3.62 (-34)***
Control 83.37 6.82
5
100
200
Exp 70.11 5.81(-14)*
Exp 63.39 4.77 (-23)**
Control 85.55 7.00
Exp 57.77 3.82(-19)*
Exp 49.22 3.50 (-31)***
Control 75.32 6.32
Exp 68.63 5.77 (-18)*
Exp 55.92 4.66 (-32)***
Control 85.66 6.66
6
100
200
Exp 75.22 6.32 (-12)*
Exp 64.48 5.11 (-24)**
Control 88.77 6.13
Exp 63.49 4.88 (-15)*
Exp 56.66 3.82 (-24)***
Control 63.34 6.00
Exp 65.55 3.82 (-23)**
Exp 57.33 4.82 (-33)***
Control 84.38 6.03
7
100
200
Exp 70.22 4.92 (-20)**
Exp 64.79 4.88 (-27)***
Control 90.13 7.34
Exp 50.00 3.81 (-21)**
Exp 42.77 3.70 (-32)***
Control 65.59 5.20
Exp 70.11 6.32 (-17)*
Exp 64.44 5.34 (-24)***
Control 86.62 7.11
8
100
200
Exp 68.13 4.87 (-24)***
Exp 66.22 5.11 (-26)***
Control 87.33 7.72
Exp 47.74 4.72 (-27)***
Exp 46.22 3.61 (-29)***
Control 63.82 4.88
Exp 68.82 5.32 (-20)**
Exp 62.11 5.88 (-28)***
Control 88.88 6.21
9
100
200
Exp 62.44 5.33 (-28)***
Exp 57.32 4.44 (-34)***
Control 93.44 6.84
Exp 53.22 4.82 (-16)*
Exp 47.33 3.55 (-25)***
Control 80.48 7.21
Exp 66.77 5.33 (-24)***
Exp 55.22 4.11 (-37)***
Control 98.08 7.08
10
11
12
13
14
100
200
Exp 80.52 6.13 (-14)**
Exp 70.54 5.92 (-24)**
Control 91.42 8.11
Exp 72.85 6.21 (-9)NS
Exp 67.38 5.28 (-16)*
Control 82.61 6.63
Exp 73.39 6.11 (-17)**
Exp 67.11 6.00 (-23)***
Control 96.12 7.22
100
200
Exp 78.23 5.39 (-14)*
Exp 66.66 4.82 (-27)***
Control 90.88 7.84
Exp 71.11 5.32 (-14)*
Exp 68.89 6.00 (-17)**
Control 78.18 6.22
Exp 78.44 5.99 (-18)**
Exp 72.66 5.33 (-24)***
Control 85.55 7.00
100
200
Exp 80.77 6.21 (-11) NS
Exp 73.41 5.82 (-19)*
Control 91.11 6.94
Exp 64.42 4.81 (-18)*
Exp 59.92 5.00 (-23)**
Control 76.66 5.23
Exp 76.63 5.81 (-10) NS
Exp 64.89 5.00 (-28)***
Control 84.11 7.00
100
200
Exp 80.27 7.00 (-11)*
Exp 68.41 5.24 (-25)***
Control 94.44 6.92
Exp 66.11 5.11 (-14)*
Exp 53.33 4.22 (-30)***
Control 73.33 6.00
Exp 77.14 6.11 (-8) NS
Exp 71.11 6.44 (-15)*
Control 76.66 5.24
100
200
Exp 80.22 7.00 (-15)*
Exp 72.99 6.33 (-22)**
Exp 64.48 5.11 (-12)*
Exp 53.44 3.90 (-27)***
Exp 60.11 5.00 (-22)**
Exp 54.44 4.71 (-28)***
123

Med Chem Res (2011) 20:36–46
45
Table 4 continued
Treatment
Conc. (lg/ml)
Formation of superoxide anionsa
Formation of hydroxyl radicalsb
Microsomal lipid peroxidationc
16
Control 86.77 5.43
Exp 76.33 5.22 (-12)*
Exp 68.83 5.11 (-21)**
Control 26.88 2.00
Exp 4.21 0.02 (-83)***
(Alloperinol) (20 lg/ml)
b
Control 68.88 5.22
Control 77.77 6.34
100
200
Exp 57.77 4.66 (-16)*
Exp 44.44 3.11 (-35)***
Control 38.44 3.11
Exp 62.22 5.33 (-20)**
Exp 53.44 5.00 (-31)***
Control 4.39 0.03
Standard drug
Exp 22.44 2.00 (-42)**s*
(Mannitol) (100 lg/ml)
Exp 2.00 0.01 (-54)***
(a-Tocopherol) (20 lg/ml)
a
Units: nmol formazone formed/min; nmole MDA/h. Each value is the mean SD of four separate observations. * P\ 0.05, ** P\ 0.01,
*** P\ 0.001, NS non significant as compared to the systems without drug treatment
Asahina T, Kashiwagi A, Nishio Y (1995) Impaired activation of
(26–35%) and microsomal lipid peroxidation (23–34%)
glucose oxidation and NADPH supply in human endothelial
inhibition in above test system as compared to that of com-
cells exposed to H2O2 in high glucose medium. Diabetes 44:
520–526
Bhatia G, Puri A, Maurya R, Yadav PP, Khan MM, Khanna AK,
pounds 10, 11 and 13. The involvement of hydroxyl free
radicals (OH•) has been found to be a major causative factor
Saxena JK (2008) Anti-dyslipidemic and antioxidant activities of
for peroxidative damage to lipoproteins, which is responsi-
different fractions of Pongamia pinnata (lin) fruits. Med Chem
ble for inhibition and progression of atherosclerosis in
Res 17:281–289
Chander R, Khanna AK, Kapoor NK (1996) Lipid lowering activity
hyperlipidemic subjects (Parthasarathy et al., 1992).
To access the antioxidant activity of pregnane deriva-
tives, we have used in vitro model of non-enzymic super-
oxide hydroxy radicals and microsomal lipid peroxidation.
The test drug showed for their inhibitory action against
microsomal lipid peroxidation in vitro by non-enzymatic
inducer. Malondialdehyde contents in both experimental
and reference tubes were estimated spectrophotometrically
by thiobarbituric acid method. The lowering of superoxide
and microsomal hydroxy ion in different analogues of
pregnane derivatives 5, 6, 7, 8, 9, 13, and 14 showed
inhibition of SOD and OH- ions is 23, 32; 24, 33; 27, 24;
26, 28; 34, 37; 25, 15; 22, 28%, respectively (Table 4). The
properties of pregnane derivatives 7, 8, 13, and 14 as anti-
oxidant and scavenger of oxygen free radicals appear to be
mediated through activity like metal ion chelators and
xanthine oxidase inhibitors.
of guggulsterone from Commiphora mukul in hyperlipemic rats.
Phytother Res 10:508–511
Das R, Kirk DN (1984) 18-Substituted steroids. Part 11. Synthesis of
3b, 16a, 18-trihydroxyandrost-5-en-17-one, a Neonatal Urinary
Metabolite and the 3, 16, 18-triacetate of its 16b-epimer. J Chem
Soc Perkin Trans I:1821–1831
Fukushima DK, Gallagher TF (1951) The action of alcoholic
potassium hydroxide on D16-20-ketosteroid. J Am Chem Soc
73:196–201
Jammart-Gregore B, Caubere P, Blank M, Grassounou JP, Advenier
C (1989) A new series of tricyclic (ary1oximino) propanolam-
ines displaying very high selective &blocking properties. J Med
Chem 32:315–320
Kaneko K, Watanabbe M, Mitsuhashi K (1973) 3b-Hydroxy-D5,16
-
pregnadien-20-one from veratrum grandiflorum. Phytochemistry
12:1509–1510
Kirk M (1993) NMR study of conformation and configuration in
C-20-substituted 5b,14b-pregnanes, 5b-pregn-14-enes and 21-
nor-5b,14b-pregnanes. Magn Reson Chem 31:17–22
Lewbart ML (1968) Synthesis of the four pairs of side-chain epoxides
epimeric at carbon 20 derived from 5b-pregnan-3a-ol. J Org
Chem 33:1695–1706
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein
measurement with folin phenol reagent. J Biol Chem 193:265–
275
Marker RE (1949) Steroidal sapogenins-173. 16-dehydropregnen-5-
ol-3-dione-12, 20 from ricogenin, a new steroidal sapogenin.
J Am Chem Soc 71:3856
Muller GP, Norton LL (1955) Ring-D reactions of 3b-acetoxy-5a-
pregn-16-ene-12, 20-dione. J Am Chem Soc 77:143–145
Neher P, Desaulles P, Vischer E, Wieland P, Wettstein A (1958)
Isolierung, Konstitution und Synthese eines neuen Steroides aus
Therefore, it is envisaged that, a drug having multifold
properties such as antioxidant, anti-diabetic and lipid
lowering activity is in great demand.
In conclusion, a series of novel pregnane derivatives
have been synthesized and some of them have been shown
to be effective anti-dyslipidemic and antioxidant agents.
Further optimization of some of the lead molecules are
currently in progress in out laboratory.
Acknowledgements The authors are grateful to the RSIC division of
the Central Drug Research Institute, Lucknow for the analytical data.
¨
Nebennieren. Uber Steroide, 155. Mitteilung Helv Chim Acta
41:1667–1692
Parthasarathy S, Steinbert D, Swiztum JL (1992) The role of oxidized
low-density lipoproteins in the pathogenesis of atherosclerosis.
Ann Rev Med 43:219
References
Rosenkranz G, Mancera O, Sondheimer F, Djerassi C (1956) Steroids
LXXXI transformation of sapogenins to androgens and estro-
gens, Beckmann rearrangement of D16-20-ketosteroids. J Org
Chem 21:520–522
Alexander CM, Londsman PB, Teatsch SM (2002) Diabetes mellitus
impaired fasting glucose, artherosclerosis factors and prevalence
of coronary heart diseases. Am J Cardiol 86:897–902
123

46
Med Chem Res (2011) 20:36–46
Ruotola G, Howard BV (2002) Dyslipidemia of the metabolic
syndrome. Curr Cardiol Rep 4:494–500
Schurr PE, Schultz JR, Parkinson TM (1972) Triton-induced hyper-
lipidemia in rats as an animal model for screening hypolipidemic
drugs. Lipids 7:68–74
Stubbs KA, Macauley MS, Vocadlo DJ (2006) A highly concise
preparation of O-deacetylated arylthioglycosides of N-acetyl-D-
glucosamine from 2 acetamido-3,4,6-tri-O-acetyl-2-deoxy-a-D-
glucopyransyl chloride and arylthiols or disulfides. Carbohydr
Res 341(10):1764–1769
Sethi A, Maurya A, Tewari V, Srivastava S, Faridi S, Bhatia G, Khan
MM, Khanna AK, Saxena JK (2007) Expeditious and convenient
synthesis of pregnanes and its glycosides as potential anti-
dyslipidemic and anti-oxidant agents. Bioorg Med Chem
15:4520–4527
Sethi A, Sudha P, Srivastava S, Naveen NK, Bhatia A, Alok K, Bhatia
G, Khan MM, Khanna AK, Saxena JK (2008) A new pregnane
glycoside from Marsdenia roylei as potential antioxidant and
anti-dyslipidemic agent. J Asian Nat Prod Res 10:1023–1028
Stehouwer CDA, Lambert J, Donker AJM, Vanhinsbergh VWM
(1997) Endothelial dysfunction and pathogenesis of diabetic
angiopathy. Cardiovasc Res 34:55–61
Wang X, Greilberger J, Ledinski G, Kager G, Paigen B, Jurgnes G
(2004) The hypolipidemic natural product Commiphora mukul
and its component guggulsterone inhibit oxidative modification
of LDL. Atherosclerosis 172:239–246
Windler EE, Kovanen PT, Chao YS, Brown MS, Havel RJ, Goldstein
JL (1980) The estradiol-stimulated lipoprotein receptor of rat
liver. A binding site that membrane mediates the uptake of rat
lipoproteins containing apoproteins B and E. J Biol Chem 255:
10464–104671
Wing DR, Robinson DF (1968) Clearing-factor lipase in adipose
tissue. A possible role of adenosine 30,50 (cyclic)-monophosphate
in the regulation of its activity. Biochem J 109:841–849
123