4alpha-methyl-24beta-ethyl-5alpha-cholesta-14,25-dien-3beta-ol and 24beta-ethylcholesta-5, 9(11), 22E-trien-3beta-ol, sterols from Clerodendrum inerme.

Article abstract of DOI:10.1016/S0031-9422(03)00146-8

From the aerial parts of Clerodendrum inerme, two new sterols (4alpha-methyl-24beta-ethyl-5alpha-cholesta-14, 25-dien-3beta-ol and 24beta-ethylcholesta-5, 9(11), 22E-trien-3beta-ol) and a new aliphatic ketone (11-pentacosanone) were isolated together with another known aliphatic ketone (6-nonacosanone) and a diterpene (clerodermic acid). The structure elucidations were based on analyses of physical and spectroscopic data.

Full text of DOI:10.1016/S0031-9422(03)00146-8

Phytochemistry 63 (2003) 415–420
www.elsevier.com/locate/phytochem
4a-Methyl-24b-ethyl-5a-cholesta-14,25-dien-3b-ol and
24b-ethylcholesta-5, 9(11), 22E-trien-3b-ol, sterols
from Clerodendrum inerme
Richa Pandey, Ram K. Verma, Subhash C. Singh, Madan M. Gupta*
Central Institute of Medicinal and Aromatic Plants, Lucknow, 226015, India
Received 6 September 2002; received in revised form 12 February 2003
Abstract
From the aerial parts of Clerodendrum inerme, two new sterols (4a-methyl-24b-ethyl-5a-cholesta-14, 25-dien-3b-ol and 24b-
ethylcholesta-5, 9(11), 22E-trien-3b-ol) and a new aliphatic ketone (11-pentacosanone) were isolated together with another known
aliphatic ketone (6-nonacosanone) and a diterpene (clerodermic acid). The structure elucidations were based on analyses of physical
and spectroscopic data.
# 2003 Elsevier Science Ltd. All rights reserved.
Keywords: Clerodendrum inerme; Verbenaceae; Sterols
1. Introduction
biosynthesis (Nes and Mckean, 1977; Benveniste, 1986)
in Clerodendrum inerme. It led to the isolation and
characterization of a new 4a-methylsterol, 4a-methyl-
24b-ethyl-5a-cholesta-14, 25-dien-3b-ol (1a) and
another new sterol, 24b-ethylcholesta-5, 9(11), 22E-
trien-3b-ol (2a) along with a new aliphatic ketone
11-pentacosanone (3), 6-nonacosanone (4) and a diter-
pene clerodermic acid (5).
Clerodendrum inerme (L.) Gaertn. (Verbenaceae) is a
common shrub that grows in India, both in the wild and
as a garden hedge. Its leaves are used as alterative, feb-
rifuge and as a substitute for Swertia chirayita. Its
leaves have been shown to possess antimicrobial activ-
ity (Prasad et al., 1995) and are reported to be cardio-
vascular system active. They also stimulate uterine
motility in rats and inhibit intestinal motility (Husain
et al., 1992). The plant contains mainly iridoids, flavo-
noids, diterpenes, sterols, triterpenes and neolignans
(Akihisa et al., 1990a; Rehman et al., 1997; Kan-
chanapoom et al., 2001).
The genus Clerodendrum is known to contain
24b-ethylsterols possessing a Á25 bond as the dominant
sterols (Akihisa et al., 1990a). 4 a-Methylsterols viz. 4a,
24, 24-trimethyl-5a-cholesta-7, 25-dien-3 b-ol and
4a-methyl-24b-ethyl-5a-cholesta-7, 25-dien-3 b-ol toge-
ther with two other common sterols have also been iso-
lated from this plant (Akihisa et al., 1990a). This paper
describes further investigation on 4a-methylsterols,
which are important intermediates in 4-demethyl sterol
2. Results and discussion
The compound 1a isolated from the fractions eluted
with hexane–ethyl acetate (95:5) mp 170–172ꢀ, [a]d+78ꢀ
possessed molecular formula C30H50O (MS) and ꢀmax
3400 and 1040 cmÀ1 for an equatorial OH group; 1640
and 880 cmÀ1 for double bond and 2940, 2860, 1460,
1380 cmÀ1 for CH, CH2 and CH3 functions (Gupta et
al., 1981). The 1H NMR spectrum displayed two signals
at ꢁ 0.89 and 0.83 for C-18 and C-19 angular methyl
groups, respectively, typical of a Á14-sterol (Kokke et
al., 1981; Dow et al., 1983). Signals corresponding to
C-21 and C-30 methyl groups appeared at ꢁ 0.92 (d) and
0.94 (d), respectively. C-15 olefinic proton appeared at ꢁ
5.17 (Kokke et al., 1981). Another double bond signals
at ꢁ 4.57 and 4.69 together with C-29 and C-26 methyls
* Corresponding author. Fax: +41-522-342666.
E-mail address: cimap@satyam.net.in (M.M. Gupta).
0031-9422/03/$ - see front matter # 2003 Elsevier Science Ltd. All rights reserved.
doi:10.1016/S0031-9422(03)00146-8

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R. Pandey et al. / Phytochemistry 63 (2003) 415–420
at ꢁ 0.82 (t) and 1.68 (s) are in good agreement with the
double bond at Á25 (Akihisa et al., 1988a). A multiplet
assignable to C-3 axial methine appeared at ꢁ 3.22
(Gupta et al., 1981). In its 1H NMR spectrum the
observed value (ꢁ 0.83) for C-19 methyl group has close
resemblance with the value calculated by Zurchers’ rule
for 5a-androstan (calculated value ꢁ 0.82) than for a
5b-androstan (calculated value ꢁ 0.95), suggesting a
5a-H in 1a (Zurcher, 1963). The side chain proton sig-
nals were closely correlated with those of 24 b-ethyl-
cholesta-5, 25-dien-3 b-ol (Akihisa et al., 1988a), which
made it possible to determine the configuration at C-24
of 1a to be b. The mass spectrum of 1a showed mole-
cular ion peak at m/z 426 (C30H50O), loosing a water
molecule at m/z 408 and a methyl group at m/z 411.
Loss of side chain (C10H19) from the molecular ion
resulted in an ion at m/z 287. Ion at m/z 218 was due to
C ring cleavage. Base peak at m/z 121 was due to the
loss of C7H13 of the side chain from the ion at m/z 218.
The 13C NMR spectrum also showed the presence of
30 carbon signals in the molecule. The DEPT spectrum
exhibited six methyls, eleven methylene and nine
methine, while the remaining four signals in the broad
band spectrum were due to the quaternary carbon
atoms. It showed six methyl group resonances at ꢁ 15.4,
15.5, 16.8, 18.0, 18.4 and 20.9. Two sets of olefinic car-
bon resonances occurred at ꢁ (121.7, 145.2) and (109.3,
151.0) corresponding to trisubstituted and disubstituted
double bonds at C14 and C25, respectively. The signal at
ꢁ 79.0 was due to the carbon bearing oxygen. These data
suggested (Kokke et al., 1981; Akihisa et al., 1988a,
1990a) the presence of a 4a-methyl and two double
bonds at Á14 and Á25.
Acetylation of 1a afforded a monoacetate 1b, ꢀmax
1733 and 1253 cmÀ1. The singlet nature of the latter
‘acetate band’ established the equatorial orientation of
the C-3 hydroxyl group (Gupta et al., 1981; Jones et al.,
1951). The C-3 methine signal was shifted downfield by
about 1.3.
Jones’ oxidation of 1a furnished a ketone 1c, in which
the C-18 and C-19 methyl groups appeared as singlet at
ꢁ 0.87 and 1.10, respectively. Its IR and UVspectra

R. Pandey et al. / Phytochemistry 63 (2003) 415–420
417
lacked absorption for an a, b-unsaturated ketone indi-
cating the absence of a double bond at C-5.
Lin et al., 1988). The signal at ꢁ 71.8 was due to the
carbon bearing oxygen.
Thus, on the basis of above spectral evidences the
structure of sterol 1a was formulated as 4a-methyl-24b-
ethyl-5a-cholesta-14, 25-dien-3b-ol.
Acetylation of 2a yielded a monoacetate 2b, ꢀmax 1735
and 1260 cmÀ1 and the singlet nature of the latter ‘ace-
tate band’ established the equatorial orientation of the
C-3 hydroxyl group (Gupta et al., 1981; Jones et al.,
1951). The C-3 methine signal was shifted to ꢁ 4.60.
Authentic cholesterol (Á5, 3b-hydroxysterol), pur-
chased from Sigma, India, was oxidised with Jones’
reagent to yield Á4-cholesten-3-one (6), an a,b-unsatu-
rated ketone. Comparison of 1H NMR and UVspectral
data of a,b-unsaturated ketone 2c, obtained after Jones’
oxidation of 2a, and 6 suggested a double bond at C-5
in 2a similar to cholesterol (1H NMR: ꢁ 6.17 (s) in 6 for
C-4 of a,b-unsaturated ketone; ꢁ 6.17(s) in 2c for C-4.
UV: 247 nm in 6; 248 nm in 2c). Proton signals for C-6
H at ꢁ 5.34 (2a) and ꢁ 5.35 (cholesterol) disappeared in
the respective oxidation products 2c and 6.
Compound 2a, mp 160–162ꢀ, [a]d À47ꢀ, was isolated
from the fractions eluted with hexane-ethyl acetate (95:
5). The EI mass spectrum of 2a exhibited a peak at m/z
410 compatible with the molecular formula C29H46O.
Other significant peaks were observed at m/z 392
[M–H2O]+, 395 [M–CH3]+, 271 [M–C10H19 (side
chain)]+, 253 [M–H2O–C10H19]+, 229 [M–C10H19–D
ring cleavage (C3H6)]+, 190 [C ring cleavage]+, 176 [C
ring cleavage]+and 138 [B ring cleavage]+ indicating
that it is a C29 sterol with three double bonds, one of
which was in the C10 side chain and the other two were
in the skeleton. Absence of double bond in the D ring
was indicated by the fragment ions at m/z 190 and 176,
resulted by C ring cleavage. Ion at m/z 138 was due to B
ring cleavage. The IR spectrum displayed the presence
of hydroxyl (3432 and 1059 cmÀ1) and olefinic (1640
cmÀ1) groups. Absorption maxima at 2939, 2865, 1458,
1376 cmÀ1 were due to the stretching and bending
vibrations of CH, CH2 and CH3 groups.
Thus, on the basis of above spectral evidences struc-
ture of compound 2a was established as 24b-ethylcho-
lesta-5, 9(11), 22E-trien-3b-ol.
Compound 3, mp 66–68ꢀ, possessed IR absorption
bands at 2910, 2855, 1705, 720 cmÀ1 for a long chain
aliphatic ketone. Molecular ion at m/z 366 suggested its
molecular formula as C25H50O. Presence of strong ions
at m/z 225, 197, 169, 141, and at 240, 184 were due to a
and b fissions, respectively indicating a carbonyl group
at C-11 (Gupta and Verma, 1991). A double rearrange-
ment ion seen at m/z 58 was characteristic of a ketone
having ꢂ H in both alkyl fragments. In its 13C NMR,
the terminal methyls appeared at ꢁ 14.1, –CH2–CO–
CH2–at ꢁ 33.9 and the carbonyl group at ꢁ 178.0. Thus,
the structure of compound 3 was assigned as 11-penta-
cosanone. This compound is being reported for the first
time in nature.
1
In the H NMR spectrum signals corresponding to
methyl groups at C-21, C-26, C-27 and C-29 appeared
at ꢁ 1.02 (d), 0.79 (d), 0.84 (d) and 0.80 (t), respectively
(Akihisa et al., 1988a, b). The olefinic protons signals at
ꢁ 5.01 (1H, H-23) and 5.19 (1H, H-22) were due to the
presence of a double bond at C-22 (Akihisa et al.,
1988b) while the signal at ꢁ 5.21 (1H, t, H-11) indicated
the presence of a double bond at C-9 (Akihisa et al.,
1990b). The angular methyl groups at C-18 and C-19
resonated at ꢁ 0.70 (s) and 1.01 (s), respectively, typical
9(11)
of a Á
sterol (Gupta et al., 1981). The signals at ꢁ
5.35 (1 H, br d, H-6) and the multiplet at ꢁ 3.52 (1H,
m, H-3) were characteristic of a Á5-3b-hydroxyl sterols
(Goswami et al., 1996). The stereochemistry at C-22
was established to be E (trans) since the 21-H3 doublet
appeared at ꢁ 1.02; whereas a Z (cis) double bond
resonates at ꢁ 0.94–0.95 (Akihisa et al., 1988a). The
configuration at C-24 was established as 24b by com-
paring side chain protons of a 24b sterol, 4a-methyl-
24b (S)-ethyl-5a-cholest-22E-en-3b-ol (Tam Ha et al.,
1982).
The 13C NMR spectrum showed 29 signals corre-
sponding to 29 carbon atoms in the molecule. The
DEPT spectrum showed the presence of six methyl,
eight methylene and eleven methine groups while the
remaining four carbon atoms existed in the quaternary
form. Resonances due to the presence of six methyl
groups were seen at ꢁ 12.0, 12.2, 19.4, 20.2, 20.8 and 21.2.
Three sets of olefinic carbon resonances occurred at ꢁ
121.7, 140.7; 129.3, 138.3 and 130.0, 137.2 corresponding
to two trisubstituted and one disubstituted double bonds
at Á5, 9(11), 22 (Akihisa et al., 1988b; Shirane et al., 1990;
Compound 4, mp 74–76ꢀ, showed IR bands at 2920,
2860, 1702, 720 cmÀ1. Molecular ion peak at m/z 422
suggested the molecular formula as C29H58O. Presence
of strong ions at m/z 351, 323, 99, 71, and 366, 337 due
to a and b fissions, respectively indicated the carbonyl
group at C-6 (Gupta and Verma, 1991). Similar to 3 a
double rearrangement ion at m/z 58 was due to the pre-
sence of ꢂ H in both alkyl fragments. This is the first
report of the occurrence of this compound in this plant
although it has been reported earlier from another nat-
ural source, Cassia auriculata (Lohar et al., 1981).
3. Experimental
3.1. General
Mps: uncorr. The 300 MHz NMR spectra were
recorded in CDCl3 with tetramethyl silane (TMS) as
internal standard. The 13C NMR (broad band and
DEPT) spectra were recorded at 75 MHz. The DEPT

418
R. Pandey et al. / Phytochemistry 63 (2003) 415–420
experiments were used to determine the multiplicities of
carbon atoms. TLC was performed on 60 F254 pre-
coated silica gel plates (Merck) and the spots were
visualized by exposure to I2 vapours or by spraying with
vanillin: sulphuric acid: ethanol (1 g: 5 ml: 95 ml)
(C-17), 79.0 (C-3), 109.3 (C-27), 121.7 (C-15), 145.2
(C-14), 151.0 (C-25); EIMS m/z (rel. int.): 426[M]+
(C30H500, 3.4), 411 [M–Me]+ (1.5), 408 [M–H2O]+
(2.5), 287 [M–C10H19 (SC)]+ (0.5), 218 [C ring clea-
vage]+(14.8), 121 [218-C7H13 of SC]+(100).
ꢀ
reagent followed by heating the plate at 110 C for 15
min.
3.5. Acetylation of 1a
3.2. Plant material
Compound 1a (0.012 g) was dissolved in pyridine (2
ml), and Ac2O (2 ml) was added. It was left overnight at
room temp. and then diluted with cold H2O (50 ml) and
extracted with Et2O (4Â50 ml). The Et2O extract was
washed successively with dil. HCl (2Â50ml), NaHCO3
soln (2Â50ml) and H2O (2Â50 ml), and dried over
Na2SO4. Crystallisation from acetone provided 1b
Aerial parts of Clerodendrum inerme were collected
from Lucknow in October 2000 and a voucher specimen
(No. CIMAP-8199) has been deposited in the herbar-
ium of this institute.
ꢀ
3.3. Extraction and isolation
(0.007 g), mp 212–214 C. IR ꢀmax cmÀ1: 2943, 2855,
1
1733, 1635, 1462, 1367, 1253 and 1027. H NMR (300
Air dried and finely powdered aerial parts of the plant
(8.2 kg) were exhaustively extracted at room tempera-
ture (25–26 ꢀC) with ethanol (10 lÂ3 times) and the
ethanolic extract was concentrated in vacuo to give a
residue. Water (500 ml) was added and it was then par-
titioned with n-hexane, chloroform and n-butanol. The
n-hexane extract (80.46 g) was subjected to column
chromatography on silica gel, 1.5 kg (60–120 mesh).
Elution was carried out in varying percentage of EtOAc
in hexane. Fractions eluted with hexane–ethyl acetate
(95:5) afforded compounds 1a and 2a. Compound 1a
(0.06 g) was obtained in crystalline form after crystal-
lization from CHCl3–CH3COCH3. Compound 2a
(0.35g) was obtained as colourless crystals from CHCl3–
CH3COCH3. Compounds 3 (0.04 g) and 4 (0.22 g) were
obtained from earlier fractions eluted with hexane–
EtOAc (95:5) and were crystallised from CHCl3-MeOH.
The fractions eluted with hexane–EtOAc (70:30) were
crystallised from hexane–EtOAc to provide 5 (0.09 g).
Clerodermic acid and 6-nonacosanone were identified
by comparison of their spectroscopic data with pub-
lished data (Achari et al., 1990; Lohar et al., 1981).
M Hz, CDCl3): ꢁ 0.80 (3H, t, J=9.0Hz, H-29), 0.83
(3H, s, H-19), 0.87 (3H, s, H-18), 0.91 (3H, d, J=8.0Hz,
H-21), 0.94 (3H, d, J=8.0 Hz, H-30), 1.66 (3H, s, H-26),
2.05 (3H, s, COOCH3), 4.50 (1H, m, H-3), 4.56 (1H, brs,
H-27), 4.68 (1H,brs, H-27), 5.17 (1H, m, H-15); EIMS
m/z: 468 [M]+ (C32H5202).
3.6. Jones’ oxidation of 1a
Compound 1a (0.015 g) was dissolved in CH3COCH3
(250 ml) and 8 N chromic acid was added dropwise with
constant shaking. Completion of the reaction was indi-
cated by the persistence of the yellow colour in the
supernatant liquid even after 10 min. The residue was
conc. to 50 ml in vacuo diluted with H2O (100 ml),
extracted with Et2O (4Â50 ml). The Et2O extract was
washed with H2O (100 ml) and dried (Na2SO4). It was
purified by preparative TLC (benzene–acetone; 95:5) to
give a viscous mass, 7 mg. Due to the small amount it
resisted crystallization. IR ꢀmax cmÀ1: 2938, 2855, 1705,
1635, 1450, 1370, 1022, 900; 1H NMR (300 MHz,
CDCl3): ꢁ 0.81 (3H, t, J=9.0 Hz, H-29), 0.87 (3H, s,
H-18), 0.92 (3H, d, J=8.0 Hz, H-21), 1.04 (3H, d,
J=8.0 Hz, H-30), 1.10 (3H, s, H-19), 1.68 (3H, s,
H-26), 4.56 (1H, br s, H-27), 4.69 (1H, d, J=2.1 Hz,
H-27), 5.18 (1H, m, H-15); EIMS m/z: 424[M]+
(C30H480).
3.4. 4ꢃ-Methyl-24ꢄ-ethyl-5ꢃ-cholesta-14, 25-dien-3ꢄ-ol
(1a)
Crystalline solid; mp 170–172ꢀ; [a]d+77ꢀ (CHCl3); IR
ꢀmax cmÀ1: 3400, 2940, 2860, 1640, 1460, 1380, 1040,
1
880; H NMR (300 MHz, CDCl3): ꢁ 0.82 (3H, t, J=9.0
3.7. 24ꢄ-Ethylcholesta-5, 9 (11), 22E-trien-3ꢄ-ol (2a)
Hz, H-29), 0.83 (3H, s, H-19), 0.89 (3H, s, H-18), 0.92
(3H, d, J=8.0 Hz, H-21), 0.94 (3H, d, J=8.0 Hz, H-30),
1.68 (3H, s, H-26), 3.22 (1H, m, H-3), 4.57 (1H, br s,
H-27), 4.69 (1H, d, J=2.1 Hz, H-27), 5.18 (1H, m,
H-15); 13C NMR (CDCl3): ꢁ 15.4 (C-29), 15.5 (C-18),
16.8 (C-30), 18.0 (C-26), 18.4 (C-19), 20.9 (C-21), 23.5
(C-11), 26.2 (C-28), 29.7 (C-6), 29.8 (C-23), 31.1 (C-2),
32.5 (C-16), 32.6 (C-7), 33.3 (C-22), 34.3 (C-8), 34.7
(C-10), 35.6 (C-20), 37.1 (C-1), 38.8 (C-12), 40.0 (C-4),
46.8 (C-13), 47.2 (C-5), 50.4 (C-24), 55.2 (C-9), 55.3
Crystalline solid; [a]dÀ47ꢀ (CHCl3); mp 158–160ꢀ; IR
ꢀmax cmÀ1 : 3432, 2939, 2865, 1640, 1458, 1376, 1244,
1
1059, 966; H NMR (300 MHz, CDCl3): ꢁ 0.70 (3H, s,
H-18), 0.79 (3H, d, J=6.3 Hz, H-26), 0.84 (3H, d,
J=6.9 Hz, H-27), 0.80 (3H, t, J=7.0 Hz, H-29), 1.01
(3H, s, H-19), 1.02 (3H, d, J=6.9 Hz, H-21), 3.52 (1H,
m, H-3), 5.01 (1H, dd, J=8.4, 15.3 Hz, H-23), 5.19 (1H,
dd, J=8.4, 15.3 Hz, H-22), 5.21 (1H, t, J=8.1 Hz, H-
11), 5.35 (1H, br d, J=4.8 Hz, H-6);13C NMR (CDCl3):

R. Pandey et al. / Phytochemistry 63 (2003) 415–420
419
ꢁ 12.0 (C-18), 12.2 (C-29), 19.4 (C-19), 20.2 (C-26), 20.8
(C-27), 21.2 (C-21), 24.3 (C-15), 25.7 (C-28), 28.7
(C-16), 28.9 (C-2), 31.6 (C-8), 31.9 (C-7), 36.5 (C-25),
37.2 (C-1), 39.7 (C-12), 40.2 (C-10), 40.3 (C-20), 40.5
(C-13), 42.3 (C-4), 50.1 (C-24), 55.9 (C-17), 56.8 (C-14),
71.8 (C-3), 121.7 (C-6), 129.3 (C-11), 130.0 (C-23), 137.2
(C-22), 138.3 (C-9), 140.7 (C-5); EIMS m/z (rel. int.):
410 [M]+ (C29H46O, 71.2), 395 [M–CH3]+(9.3), 392
Cholesterol (0.05 g) was oxidized as above to yield an
1
a, b-unsaturated ketone; UV lmax 247 nm ; H NMR
(300 MHz, CDCl3): ꢁ 6.17 (1H, s, H-4).
3.10. 1-Pentacosanone (3)
ꢀ
Solid; mp 66–68 C; IR ꢀmax cmÀ1: 2910, 2855, 1705,
1
1460, 1380, 720; H NMR (300 MHz, CDCl3): ꢁ 0.88
[M–H2O]+(34.0),
367
[M–C3H7]+(18.9),
271
(6H, t, J=7.5 Hz, CH3), 1.25 (m,-(CH2)n-), 1.63 (m),
2.35 (4H, t, J=7.5 Hz,–CH2CO–); 13C NMR (CDCl3): ꢁ
14.1 (C-1, C-25 ), 22.7 (C-2 or C-24), 24.7 (C-2 or C-24),
29.1–31.9 (C-3 to C-9, C-13 to C-23), 33.9 (C-10, C-12),
178.0 (C-11); EIMS m/z (rel. int.): 366 [M]+ (C25H50O,
4.0), 240 [M–C9H18]+ (9.0), 225 [M–C10H21]+ (5.7), 197
[M–C11H21O]+ (2.5), 184 [M-C13H26]+ (22.1), 169
[M–C14H29]+ (5.1), 141 [M–C15H29O]+ (7.4), 58
[C4H10]+ (8.5), 57 [C4H9]+ (60.5), 43 [C3H7]+ (100).
[M–C10H19 (SC)]+(59.2), 253 [M–H2O–SC]+(100), 229
[M–SC–C3H6 (D ring cleavage)]+(16.4), 211 [M–H2O–
SC–C3H6]+(34.7), 190 [M–SC–C6H9 (C ring clea-
vage)]+(2.4), 176 [M–SC–C7H11]+(9.8), 172 [M–H2O–
SC–C6H9]+(21.1), 158 [M–H2O–SC–C7H11]+(63.6), 138
[M–SC–C10H13 (B ring cleavage)]+(14.2), 120 [M–H2O–
C20H32]+(25.4).
3.8. Acetylation of 2a
Compound 2a (0.050g) was acetylated in the same
manner as compound 1a. When crystallizated from
acetone it gave crystals (0.025 g); mp 136–138ꢀ. IR ꢀmax
cmÀ1 : 2943, 2865, 1735, 1640, 1460, 1366, 1260, 1036,
Acknowledgements
We are thankful to Director, CIMAP, for keen inter-
est during the course of work and RSIC unit, Central
Drug Research Institute, Lucknow for providing the
spectral analysis.
1
969; H NMR (300 MHz, CDCl3): ꢁ 0.70 (3H, s, H-18),
0.79 (3H, d, J=6.3 Hz, H-26), 0.84 (3H, d, J=6.9 Hz,
H-27), 0.80 (3H, t, J=7.0 Hz, H-29), 1.02 (3H, s, H-19),
1.03 (3H, d, J=6.9 Hz, H-21), 2.03 (3H, s, COOCH3),
4.60 (1H, m, H-3), 5.03 (1H, dd, J=8.4, 15.3 Hz, H-23),
5.18 (1H, dd, J=8.4, 15.3 Hz, H-22), 5.20 (1H, t, J=8.1
Hz, H-11), 5.38 (1H, br d, J=4.8 Hz, H-6); 13C NMR
(CDCl3): ꢁ 12.0 (C-18), 12.2 (C-29), 19.3 (C-19), 20.2
(C-26), 20.8 (C-27), 21.2 (C-21), 24.3 (C-15), 25.7
(C-28), 28.7 (C-16), 28.9 (C-2), 31.6 (C-8), 31.9 (C-7),
36.6 (C-25), 37.0 (C-1), 39.6 (C-12), 40.2 (C-10), 40.3
(C-20), 40.5 (C-13), 42.2 (C-4), 50.0 (C-24), 55.9 (C-17),
56.8 (C-14), 74.0 (C-3), 122.6 (C-6), 129.3 (C-11), 130.0
(C-23), 137.2 (C-22), 138.3 (C-9), 139.6 (C-5); EIMS m/z
(rel. int.): 452 [M]+ (C31H48O2, absent), 392
[M–CH3COOH]+(85.9), 377 [M–AcOH–Me]+(5.7),
253 [M–AcOH–C10H19 (SC)]+(37.5), 211 [M–AcOH–
SC–C3H6 (D ring cleavage)]+ (4.8).
References
Achari, B., Chaudhari, C., Saha, C.R., Dutta, P.K., Pakrashi, S.C.,
1990. A deodane diterpene and other constituents of clerodentron
inerme. Phytochemistry 29, 3671–3673.
Akihisa, T., Ghosh, P., Thakur, S., Nagata, H., Tamura, T., Matsu-
moto, T., 1990a. 24,24-Dimethyl-25-dehydrolophenol, a 4a-methyl-
sterol from Clerodendrum inerme. Phytochemistry 29, 1639–1641.
Akihisa, T., Ghosh, P., Thakur, S., Oshikiri, S., Tamura, T., Matsu-
moto, T., 1988a. 24b-Methylcholesta-5, 22E, 25-trien-3b-ol and
24a-ethyl-5a-cholest-22E-en-3b-ol from Clerodendrum fragrans.
Phytochemistry 27, 241–244.
Akihisa, T., Matsubara, Y., Ghosh, P., Thakur, S., Shimizu, N.,
Tamura, T., Matsumoto, T., 1988b. The 24a-and 24b-epimers of
24-ethylcholesta-5, 22-dien-3b-ol in two Clerodendrum species. Phy-
tochemistry 27, 1169–1172.
Akihisa, T., Tamura, T., Matsumoto, T., Kokke, W.C.M.C., Ghosh,
P., Thakur, S., 1990b. (22Z, 24S)-Stigmasta-5, 22, 25-trien-3b-ol and
other novel sterols from Clerodendrum scandens: first report of the
isolation of a cis-Á22-unsaturated sterol from a higher plant. Jour-
nal of Chemical Society Perkin Transcription India, 2213–2218.
Benveniste, P., 1986. Sterol biosynthesis. Annual Review of Plant
Physiology 37, 275–308.
3.9. Jones’ oxidation of 2a
Compound 2a (0.050 g) was oxidized using Jones
reagent in a similar manner as compound 1a. Crystal-
lization from methanol yielded a solid (0.020 g); mp
146–148ꢀ. UV lmax 248 nm (a, b unsaturated ketone);
IR ꢀmax cmÀ1 : 2959, 2870, 1683, 1638, 1457, 1379, 1224,
Dow, W.C., Gebreyesus, T., Popov, S., Carlson, R.M.K., Djerassi, C.,
1996. Marine 4-methyl sterols: Synthesis of C-24 epimers of 4a, 24-
dimethyl-5a-cholestan-3b-ol and 360 MHz 1H NMR comparisons
to the Natural product from Plexaura homomalla. Steroids 42, 217–
230.
1
968; H NMR (300 MHz, CDCl3): ꢁ 0.74 (3H, s, H-18),
0.79 (3H, d, J=6.3 Hz, H-26), 0.84 (3H, d, J=6.9 Hz,
H-27), 0.80 (3H, t, J=7.0 Hz, H-29), 1.04 (3H, d, J=6.9
Hz, H-21), 1.17 (3H, s, H-19), 5.04 (1H, dd, J=8.4, 15.3
Hz, H-23 ), 5.14 (1H, dd, J=8.4, 15.3 Hz, H-22), 5.20
(1H, t, J=8.1 Hz, H-11), 6.17 (1H, s, H-4); EIMS m/z :
408 [M]+ (C29H440).
Goswami, P., Kotoky, J., Chen, Z.N., Lu, Y., 1996. A sterol glycoside
from leaves of Clerodendron colebrookianum. Phytochemistry 41,
279–281.
Gupta, M.M., Lal, R.N., Shukla, Y.N., 1981. 5a-Stigmast-9(11)-en-
3b-ol, a sterol from Costus speciosus roots. Phytochemistry 20,
2557–2559.

420
R. Pandey et al. / Phytochemistry 63 (2003) 415–420
Gupta, M.M., Verma, R.K., 1991. Lipid constituents of Cissus quad-
rangularis. Phytochemistry 30, 875–878.
Nes, W.R., Mckean, M.L., 1977. Biochemistry of Steroids and Other
Isopentenoids. University Park Press, Baltimore.
Husain, A., Virmani, O.P., Popli, S.P., Misra, L.N., Gupta, M.M.,
Srivastava, G.N., Abraham, Z., Singh, A.K., 1992. Dictionary of
Indian Medicinal Plants. CIMAP, Lucknow. pp. 142–143.
Jones, R.N., Humphries, P., Herling, F., Dobriner, K., 1951. Steroid
metabolism. X. The effects of stereochemical configuration at posi-
tions 3 and 5 on the infrared spectra of 3-acetoxy steroids. Journal
of American Chemical Society 73, 3215–3220.
Prasad, V., Srivastava, S., Varsha, Verma, H.N., 1995. Two basic
proteins isolated from Clerodendrum inerme Gaertn. are inducers of
systemic antiviral resistance in susceptible plants. Plant Science 110,
73–82.
Rehman, A.U., Begum, S., Saied, S., Choudhary, M.I., Akhtar, F.,
1997. A steroidal glycoside from Clerodendron inerme. Phytochem-
istry 45, 1721–1722.
Kanchanapoom, T., Kasai, R., Chumsri, P., Hiraga, Y., Yamasaki,
K., 2001. Megastigmane and iridoid glucosides from Clerodendrum
inerme. Phytochemistry 58, 333–336.
Shirane, N., Murabayashi, A., Masuko, M., Uomori, A., Yoshimura,
Y., Seo, S., Uchida, K., Takeda, K., 1990. Effect of ergosterol bio-
synthesis of a fungicide, SSF-109, in Botrytis cinerea. Phytochem-
istry 29, 2513–2520.
Kokke, W.C.M.C., Fenical, W., Djerassi, C., 1981. Sterols with unu-
sual nuclear unsaturation from three cultured marine dino-
flagellates. Phytochemistry 20, 127–134.
Tam Ha, T.B., Kokke, W.C.M.C., Djerassi, C., 1982. Minor sterols of
marine invertebrates 37. Isolation of novel coprostanols and 4a-
methyl sterols from the tunicate Ascidia nigra. Steroids 40, 433–453.
Zurcher, R.F., 1963. Protonenresonanzspektroskopie und Ster-
oidstruktur II. Die Lage der C-18-und C-19-Methylsignale in
Abhangigkeit von den Substituenten am Steroidgerust. Helvetica
Chimica Acta 46, 2054–2088.
Lin, L.J., Shiao, M.S., Yeh, S.F., 1988. Triterpenes from Ganoderma
lucidum. Phytochemistry 27, 2269–2271.
Lohar, D.R., Garg, S.P., Chawan, D.D., 1981. Phytochemical studies
on Indian medicinal plants: nonacosan-6-one from Cassia auriculata
Linn. Journal of Indian Chemical Society 58, 989–991.