MAGNETIC RESONANCE IN CHEMISTRY, VOL.

34, 777-781 (1996)

'3C NMR Assignments of D :C-Friedours-7-ene

Derivatives. Evidence of an Abnormal Methyl
Group Chemical Shift

Carlos M. Cerda-Garcia-Rojas, Hector H. Hernandez-Vidal and Pedro Joseph-Nathan*

Departamento de Quimica del Centro de Investigacih y de Estudios Avanzados, Instituto Politbcnico Nacional, Apartado
14-740, Mexico, D. F. 07000,Mexico

The I3C NMR assignments of four I):C-fnedours-7-ene derivatives were carried out OR the basis of 2D shift-
correlated NMR techniques (HETCOR and FLOCK), revealing that the signal for CH,-28 appears always very
deshielded (cu. S = 38.0). The conformation of the triterpene skeleton, obtained by MMX calculations, is compared
with that of multiflor-'l-ene.

KBY WORDS NMR; 13CNMR: D : C-friedours-7-ene derivatives;triterpenes; abnormal chemical shift; conformation; MMX
calculations

INTRODUCTION

D : C-Friedours-7-ene derivatives constitute a particular

group of triterpenes which have been isolated from
diverse species such as Acronychia baueri,' Tapeino-
sperma pseudojambosa,2 Schinus terebenthif~lius,~ Taber-
naemontana longipes? Dauidsonia pruriens' and other
plants. As indicated by the name, their structures
possess a rearranged ursane skeleton in which the
methyl group at C-14 is migrated to C-13 and the
methyl group at C-8 is migrated to C-14.6 During the
phytochemical study of plants belonging to the genus
Stevia, we isolated D :C-friedours-7-en-31-01acetate (l),
also named bauerenyl a ~ e t a t e , ' . ~from
. ~ the roots of S.
subpubescens var. intermedia' and S. salicifolia.
An inspection of the reported 'H NMR assignments
for the methyl groups of the alcohol 2' revealed some
differences with respect to our assignments. Further-
more, a search for I3C NMR data for D :C-friedours-7-
ene derivatives showed that only a tentative assignment
has been described for 1,7which was made by compari-
son with the data for other triterpenes. The DEPT spec-
trum of 1 quickly revealed an abnormally deshielded
signal for a methyl group (6 = 38.01, which was erron-
eously ascribed to a CH in the previous work.' A search
using the BASF 13C NMR data bank, which contained
over 58 000 reference confirmed the abnor-
mality of this shift, since there is no precedent for a
methyl group attached to a carbocyclic structure at
such low chemical shifts. These facts prompted us to
carry out a detailed study of the 13C NMR spectra of
D :C-friedours-7-enes,which is reported here.
* Author to whom correspondence should be addressed.
1 : R = P-OAc, a-H
2 : R = P-OH, a-H
3: R=O
4 : R=H2
RESULTS A N D DISCUSSION
The identity of acetate 1 was confirmed by comparing
the physical and spectral data for the molecule with lit-
erature value^,'.^ including a single-crystal x-ray diffrac-
tion analysis, which showed excellent agreement with
the reported data? Alkaline hydrolysis of 1 afforded
alcohol 2,' which upon oxidation with CrO, in pyridine
gave ketone 3.3*'1Alternatively, reduction of 1 with
lithium in ethylenediamine gave D : C-friedours-7-ene
(4).5
The NMR study was started with ketone 3 because
this compound showed better solubility in CDCl, than
the other three substances. The 13C chemical shifts of 3
were fully assigned on the basis of its DEPT spectrum,
its one-bond 13C-'H correlation contour plot
(HETCOR) and its long-range (geminal and vicinal)

CCC 0749-1 58 1/96/100777-05 Received 20 March 1996

0 1996 by John Wiley & Sons, Ltd. Accepted (revised) 17 May 1996
118 C. M. CERDA-GARCfA-ROJAS, H. H. HERNANDEZ-VIDAL AND P. JOSEPH-NATHAN

',C-'H correlation diagram obtained with the FLOCK
pulse sequence." The data are summarized in Table 1.
The I3C NMR spectrum of 3 showed only 29 lines,
while the DEPT trace for all protonated carbons dis-
played the expected 23 lines. Therefore, the missing
signal corresponds to a quaternary carbon, as evidenced
by an APT spectrum obtained with z = 4 ms, which
allowed us to observe the seven quaternary carbon
signals, thus revealing that the line at 6 = 32.03, assign-
ed to a methine (C-20), also contained a signal for a
quaternary carbon (C-17).
Although the I3C signals of rings A and B can easily
be assigned by comparison with the reported data for
compounds with similar molecular frameworks,' the
3314
two-and three-bond correlations shown in Table 1 con-
firmed all the assignments. For example, the signal of
the quaternary carbon C-4 showed correlations with
H-5, CH,-23 and CH3-24, while the signal of methine
carbon C-5 showed correlations with CH,-23, CH,-24
and CH,-25 (see Table 1). Even though the protons of
CH,-23 and CH3-28 have the same chemical shift
(6 = 1.04), the long-range correlations of each methyl
group can be clearly distinguished, assuming that the
carbons showing a correlation with CH,-23 should also
have a correlation with CH3-24, which in turn appears
at 6 = 1.11.
The assignments of the I3C signals of rings C, D and
E were carried out according to the following pro-
cedure. Inspection of structure 3 revealed that the
hydrogens of the methyl groups (CH,-26 to CH,-30)
can in each case be long-range coupled (through two
and three bonds) with several 13C atoms bearing differ-
ent numbers of hydrogens, as is known from the DEPT
and APT spectra. For example, CH,-28 is the only
methyl group that can be long-range coupled with two
methylene carbons (C-16 and C-22), a quaternary
carbon (C-17) and a methine carbon (C-18); CH3-30 is
the only methyl group that can be long-range coupled
with a methylene carbon (C-21) and two methine
carbons (C-19 and C-20); and so forth. Since each
methyl signal showed the expected pattern of long-
range correlations in the FLOCK diagram, the methyl
hydrogen assignments were achieved as shown in Table
1. This information also allowed the unambiguous
assignment of C-12, (2-15, C-17, C-18 and C-21 and the
assignment, although not individually, of the pairs C-13
and (2-14, C-16 and (2-22, and C-19 and C-20. The
remaining CH, signal was ascribed to C-11. Addi-
tionally, the HETCOR correlations individually assign-
ed the signals for the methyl carbons C-26 to C-30.
The distinction between C-13 and C-14 was achieved
when the 'H coupled 13C NMR spectrum of 3 was

Table 1. 13C NMR data., 'H chemical shifts and long-range correlations for
D :C-friedours-7-en-3-one (3)
Long-range correlations'
Position 6°C 6' H C--CH C--C-c--H

1 38.31 t 1.98; 1.43b H-25

2 34.89 t 2.75; 2.25 H-1
3 21 6.64 s -
4 47.77 s - H-5; H-23; H-24
5 52.17 d 1.72 H-23; H-24; H-25
6 24.57 t 2.13; 2.10 H-5; H-7
7 11 6.43 d 5.51
8 145.46 s -

9 47.86 d 2.30 H-7; H-25

10 35.28 s - H-5; H-25
11 17.00 t 1 .55b
12 32.40 t 1.52b H-27
13 37.77 s - H-27; H-18 H-26
14 41.40 s - H-26 H-27
15 28.96 t 1.55b;1.46b H-26
16 37.68 t 1.51b; 1.1gb H-28
17 32.03 s - H-28; H-18
18 54.99 d 1.30 H-27; H-28
19 35.39 d 1.16b H-29; H-18 H-30
20 32.03 d 1 .56b H-30 H-29
21 29.22 t 1 .2gb H-30
22 31.56 t 1 .57b; 1.18b H-18; H-28
23 24.50 g t .04 H-24
24 21.45 g 1.11 H-23; H-5
25 12.66 q 1.01 H-5
26 23.79 q 1.03
27 22.72 q 0.95 H-18
28 37.99 q 1.04
29 25.65 q 1.05 H-18
30 22.52 q 0.90
a Measured in CDCI, at 75.4 MHz using TMS as internal standard, multiplicities
from DEPT for protonated carbons and from APT ( r = 4 ms) for quaternary carbons.
Measured from the HETCOR correlations.
'Obtained with the FLOCK pulse sequence.
 13C NMR OF D :C-FRIEDOURS-7-ENES 719

compared with that obtained under single-frequency at 7.5 Hz and therefore the observed correlation corre-
irradiation of the vinylic proton (H-7). The signal for sponds to the coupling between H-18 and C-22.
C- 14 lost the corresponding vicinal coupling constant The assignments of the signals for D : C-friedours-
H(7)-C-C-C(14), while the signal for C-13 remained 7-en-3B-01 acetate (l),D :C-friedours-7-en-3B-01(2) and
unchanged. Since the assignment of H-20 followed from D :C-friedours-7-ene (4) followed from comparison with
a homonuclear irradiation of the CH,-30 doublet, the the I3C NMR spectrum of 3 on consideration of the
distinction between C-19 and C-20 became readily corresponding substituent effects at C-3. DEPT, APT
evident in the HETCOR contour plot. The distinction (z = 4 ms) and HETCOR spectra were also obtained for
between C-16 and C-22 was carried out from the fol- these three substances, whose assignments are listed in
lowing information. Although H-18 is three bonds away Table 2. According to these data, in addition to the
from either (2-16 or C-22, H-18 showed a FLOCK signal for C-28, the signals for C-16, C-20 and C-22 of
correlation with only one of these carbon atoms. Three- acetate 1 were incorrectly ascribed in the previous
bond 13C-'H coupling constants depend on the
H-C-C-I3C dihedral angle according to the The unambiguous assignments of the methyl proton
Karplus-type equation parametrized by Aydin and signals of 1, 2 and 4 (Table 3), were based on the corre-
Gunther.' The calculated dihedral angles H(1 8 w - lations observed in the HETCOR spectra as well as the
C-I3C(16) and H(18)--C-C-13C(22) are 72" and multiplicity of the signals (s or d). According to these
162", respectively, in the minimum energy conformation results, the previous 'H NMR assignment5 for the
of ketone 3, as obtained by MMX calculations, which is methyl groups CH,-27 and CH,-29 of 2 must be cor-
very similar to that found in the solid state for D :C- rected, since they were interchanged. Note that the 'H
friedours-7-en-3p-01 acetate (l)."The angle values (72" chemical shifts of the gem-dimethyl group are reversed
and 162") correspond to coupling constants of 0.8 and on going from 1 to 2.
8.7 Hz, respectively. In the FLOCK experiment, the The minimum energy conformation of D : C -
long-range coupling constant value [J(NXH)] was set friedours-7-ene (4), obtained by MMX calculations
[EM,, = 78.9 kcal mol-' (1 kcal = 4.184 kJ)], revealed
that rings D and E have chair and boat conformations,
respectively. As shown in Fig. l(a), there is a complete
lack of interactions between C-28 and syn-diaxial hydro-
Table 2. ''C NMR data' for D : C-friedours- gens on y-carbon atoms. Therefore, there is virtually
7-enes 1 , 2 and 4 no shielding y-effect, which could be a reason for the
Carbon l b 2 4 unusual chemical shift of C-28. We also compared the
1 36.63 t 36.92 t 38.78 t
minimum energy conformation of D : C-friedours-7-ene
2 24.26 t 27.74 t 19.14 t (4) with that recently reported'" for multiflor-7-ene (5).
3 81.19 d 79.21 d 42.45 t As shown in Fig. 1, rings A, B and C have the same
4 37.85 s 38.88 s 33.13 s conformation in both substances, whereas rings D and
5 50.68 d 50.48 d 51.21 d E show significant differences. In multiflor-7-ene (5),
6 24.04 t 24.19 t 24.64 t E M , , = 78.4 kcal mol-' (lit.'" E M , , = 78.6 kcal rnol-'),
7 116.34 d 116.49 d 1 16.56 d rings D and E display the boat conformation and C-28
8 145.50 s 145.31 s 145.55 s shifts to a less abnormal value14 (6 = 30.95). According
9 48.27 d 48.29 d 48.45 d to the MMX calculations, C-28 in 5 should experience
10 35.16 s 35.23 s 35.45 s one strong y-interaction and one weak y-interaction
11 16.91 t 16.86 t 16.88 t
12 32.53 t 32.49 t 32.62 t
with H-21B and H-l5/?, respectively, corresponding to
13 37.85 s 37.76 s 37.81 s the calculated MMX distances, which are 2.76 and 3.18
14 41.33 s 41.26 s 41.34 s respectively.
15 28.98 t 28.91 t 28.94 t In other molecules, such as taraxasteryl acetate,16
16 37.79 t 37.71 t 37.81 t C-28 appears at 6 = 19.49 regardless of the chair con-
17 32.09 s 32.06 s 32.13 s formation of ring D and the boat conformation of ring
18 55.06 d 54.97 d 55.01 d E. However, the orientation of C-28 in taraxasteryl
19 35.45 d 35.39 d 35.41 d acetate remains axial with respect to ring D, as can be
20 32.14 d 32.06 d 32.13 d assumed from the x-ray structure of taraxasterol.I6 This
21 29.29 t 29.26 t 29.30 t
22 31.61 t 31.55 t 31.61 t 3029
23 27.55 q 27.56 q 32.95 q \.:
24
25
15.81 q
13.07 q
14.66 q
12.99 q
21.27 q
12.97 q
27 li
26 23.69 q 23.65 q 23.73 q
27 22.70 q 22.67 q 22.72 q
28 38.02 q 37.99 q 38.03 q
29 25.68 q 25.65 q 25.66 q
30 22.54 q 22.55 q 22.57 q
.Chemical shifts measured in CDCI, at 75.4
MHz using TMS as internal standard. Multiplic-
ities from DEPT for protonated carbons and
from APT ( T = 4 ms) for quaternary carbons.
b6(CH,COO) : 170.85 and 21.26. 5
780 C. M. CERDA-GARCfA-ROJAS, H. H. HERNANDEZ-VIDAL AND P. JOSEPH-NATHAN

a previous c~llection'~ (1 kg) were treated with meth-

Table 3. 'H NMR chemical shifts' anol, affording an amorphous precipitate (1.1 g) which
for the methyl groups of 1, 2 was removed by filtration and chromatographed on a
and 4
silica gel column, eluting with hexane and increasing the
Methyl 1 2 4 polarity with CH,C12 . Those fractions eluted with
23 s 0.85 0.96 0.84 hexane-CH,Cl, (9: 1) gave a solid which was rec-
24 s 0.93 0.86 0.88 rystallized from CH,Cl,-hexane to yield white prisms
25 s 0.77 0.74 0.74 (220 mg) of l,'s4 m.p. 300-301 "C (lit.' 293-294 "C); [a],,
26 s 1.00 1.00 1.00 - 3.2 (c 2.5, CHCl,) (lit.' [ale - 3.7). The identity of
27 s 0.94 0.94 0.96 this substance was confirmed by x-ray diffraction
28 s 1.04 1.04 1.04 a n a l ~ s i s .EIMS,
~ 20 eV: m/z (relative intensity (%))
29 d (7 Hz) 1.05 1.05 1.05 [MI' 468 (25), 453 (lo), 301 (9), 290 (21), 289 (loo),230
30 d (7 Hz) 0.91 0.90 0.90
(12), 229 (60), 205 (17), 123 (ll),109 (14).
a Measured in CDCI, at 300 MHz using
TMS as internal standard. D : C-Friedours-7-en-3fl-01 (2). A solution of 1 (405 mg) in
CH,Cl, (40 ml) and THF (60 ml) was treated with a
solution of KOH (360 mg) in MeOH-H,O (99: 1) (40
orientation favors two strong y-interactions with H-13P ml). The mixture was refluxed for 8 h, poured over ice-
and H-15p. In contrast, the orientation of C-28 in D : C- water and extracted with CHCl, . The organic layer was
friedours-7-enes 1-4 is equatorial with respect to ring D. washed with water, dried, filtered and evaporated,
giving a residue which was chromatographed on a silica
gel column, eluting with CH,Cl,. The first fractions
EXPERIMENTAL gave recovered 1 (36 mg, 9%) and the last fractions gave
a solid which was recrystallized from CH,CI,-hexane,
yielding 21,3,5(306 mg, 83%) as white needles, m.p. 205-
Compounds 206 "C (lit.' 207-208 "C).
Isolation of D :C-friedours-7-en-3fl-01acetate (1). The hexane D : C-Friedours-7-en-3-one (3). A solution of 2 (128 mg) in
extracts (20 g) of the roots of Steuia salicifolia Cav. from CH2Cl, (6 ml) was treated with a solution of CrO, (180
mg) in pyridine (0.3 ml) and CH2Cl, (4.5 ml). The
mixture was stirred for 15 min, poured over ica- water
and extracted with CHCl,. The organic layer was
washed with dilute HC1, water, aqueous NaHCO, and
H 2 0 , dried, filtered and evaporated. The residue was
chromatographed on a silica gel column, eluting with
hexane-CH,Cl, (9: 1). Fractions 8-30 (70 ml each), on
crystallization from CH,Cl,-hexane, afforded 3, (1 14
mg, 89%) as white needles, m.p. 239-240°C (lit.,
240 "C).

D :C-Friedours-7-ene (4). This was prepared as described

previ~usly.~M.p. 243-244 "C (lit. 225-228 0C,5
247 OC").

NMR measurements

'H and 13C NMR spectra were measured on a Varian

Figure 1. Minimum energy conformations of (a) D : C-friedours-
7-ene (4) and (b) multiflor-7-ene (5). (c) Comparison between
the conformations of 4 (solid lines) and 5 (dashed lines). The
hydrogen atoms have been omitted for clarity.
XL-300GS spectrometer operating at 300 and 75.4
MHz, respectively, from CDCI, solutions with TMS as
the internal standard. The probe temperature was 23 "C.
About 80 mg of 3 were dissolved in 0.7 ml of CDCI, to
obtain the one-bond 'H-I3C shift-correlated spectrum
(HETCOR) and the long-range 'H-I3C shift-correlated
spectrum (FLOCK). In each case 1024 data points were
collected. The HETCOR spectrum was obtained using a
spectral width of 2333.2 Hz infl and 10 775.9 Hz inf, .
A total of 128 time increments were collected with 128
transients per increment. The relaxation delay was 1 s.
In the FLOCK experiment, the long-range coupling
constant value [J(NXH)] was set at 7.5 Hz. Thefl spec-
tral width was 2243.7 Hz and thef, spectral width was
5672.2 Hz. A total of 320 time increments were collected
with 128 transients per increment. The relaxation delay
 N M R O F D :C-FRIEDOURS-7-ENES
was 1 s, A1 = 0.0667 s and A2 = 0.0333 s. A value of
lJCH = 130 Hz was assumed in calculating the delay z
for the BIRD pulses.
MMX calculations
were generated using MMX force-
field calculations which is a derived version of the MM2
program developed by N. L. Allinger (QCPE 395), Uni-
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Acknowledgements
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