Glycosylation of triterpene alcohols and acids of the lupane and A-secolupane series

Article abstract of DOI:10.1135/cccc19971776

A series of 3- and 28-glucosides and glucosyl esters of betulinic acid (1a), 28-hydroxy-3,4-secolupa-4(23),20(29)-dien-3-oic acid (22a), dimethyl ester of 28-hydroxy-2,3-secolup-20(29)-en-2,3-dioic acid (43a), their 20(29)-dihydro derivatives (1b, 22b, 43b) and several other triterpenes of the lupane (12a, 12b) and 3,4-secolupane series (18a, 18b, 32a) has been prepared by reaction of tetra-O-acetyl-α-D-glucopyranosyl bromide in acetonitrile in the presence of mercury(II) cyanide and subsequent deacetylation of the obtained tetra-O-acetyl-β-D-glucopyranosyl derivatives. In several cases attempted glucosylation in the presence of silver silicate afforded predominantly the corresponding 1,2-orthoacetates of α-D-glucopyranose.

Full text of DOI:10.1135/cccc19971776

1776
Klinotova et al.:
GLYCOSYLATION OF TRITERPENE ALCOHOLS AND ACIDS
OF THE LUPANE AND A-SECOLUPANE SERIES*
a1
a2
a1
b1
Eva KLINOTOVA , Vaclav KRECEK , Jiri KLINOT , Magdalena ENDOVA ,
Jana EISENREICHOVA , Milos BUDESINSKYb2 and Martin STICHA
a
a3
a Department of Organic Chemistry, Charles University, 128 40 Prague 2, Czech Republic;
e-mail: 1 klinot@prfdec.natur.cuni.cz, 2 krecek@prfdec.natur.cuni.cz, 3 sticha@prfdec.natur.cuni.cz
b Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic,
166 10 Prague 6, Czech Republic; e-mail: 1endova@uochb.cas.cz, 2 budesinsky@uochb.cas.cz
Received July 15,1997
Accepted September 10, 1997
A series of 3- and 28-glucosides and glucosyl esters of betulinic acid (1a), 28-hydroxy-3,4-secolupa-
4(23),20(29)-dien-3-oic acid (22a), dimethyl ester of 28-hydroxy-2,3-secolup-20(29)-en-2,3-dioic
acid (43a), their 20(29)-dihydro derivatives (1b, 22b, 43b) and several other triterpenes of the lupane
(12a, 12b) and 3,4-secolupane series (18a, 18b, 32a) has been prepared by reaction of tetra-O-acetyl-
α-D-glucopyranosyl bromide in acetonitrile in the presence of mercury(II) cyanide and subsequent
deacetylation of the obtained tetra-O-acetyl-β-D-glucopyranosyl derivatives. In several cases at-
tempted glucosylation in the presence of silver silicate afforded predominantly the corresponding 1,2-
orthoacetates of α-D-glucopyranose.
Key words: Lupane; 3,4-Secolupane; 2,3-Secolupane; Betulinic acid; Glucoside; 1H and 13C NMR
spectra.
The recently found biological activity of betulinic acid (1a), its derivatives, and other
triterpenoids with lupane and A-secolupane skeleton (see e.g. refs2–9 and references
therein) aroused an increased interest in the preparation of their derivatives (inter alia
glycosides) that might exhibit biological activity. Glycosides of some lupane deriva-
tives have been prepared already earlier (see refs10,11 and references therein).
In the present paper we describe the preparation of glucosides of less common triter-
penic compounds (A-secoacids and other A-secolupane derivatives) and of betulinic
acid. Glucosides were prepared from aglycons of two parallel series: compounds with
an isopropenyl side chain in position 19 (series a; 20(29)-lupene derivatives) and the
analogous dihydro derivatives with an isopropyl group in the position 19 (series b;
lupane derivatives).
* Part CVII in the series Triterpenes; Part CVI: see ref.1.
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

Triterpenes
1777
Except the common compounds12 (betulinic acid (1a), dihydrobetulinic acid (1b) and
their derivatives 2a, 2b, 3a and 3b), the starting aglycones were prepared from the
known13 28-acetoxy-20(29)-lupen-3-one (13a) and 28-acetoxylupan-3-one (13b). The
preparation of 2,3-secodiacids 42a and 42b and their methyl esters 43a and 43b has
already been published13, the 3,4-secolupane derivatives were synthesized by proce-
dures described in analogous cases14,15: ketones 13a and 13b were converted into oximes
14a and 14b which on Beckmann rearrangement afforded acetoxy nitriles 19a and 19b
(compounds 14a, 14b, 19a and 19b are described in the literature16). Mild alkaline
hydrolysis of acetoxy nitriles gave 28-hydroxy nitriles 18a and 18b, hydrolysis with
10% sodium hydroxide in boiling ethylene glycol furnished 3,4-secoacids 22a and 22b.
These were further converted into methyl esters 23a and 23b, acetates 24a and 24b and
methyl ester acetates 25a and 25b. Reduction of methyl ester acetate 25a with lithium
aluminium hydride afforded diol 32a, characterized as diacetate 33a (the compounds
23b, 32a and 33a have been obtained earlier17–19 by other procedures). An attempted
partial acetylation of diol 32a with acetic anhydride in pyridine at –30 °C gave a mix-
ture of diacetate 33a, diol 32a and monoacetates 34a and 35a. The isomeric monoace-
tates 34a and 35a were obtained only as a 1 : 1 mixture (1H NMR spectrum) and
resisted attempted separation by chromatography or crystallization.
Of the known glycosidation procedures20 for the preparation of glucoside tetraace-
tates, we tried the Koenigs–Knorr reaction with tetra-O-acetyl-α-D-glucopyranosyl bro-
mide in the presence of silver silicate for the reaction of ketone 12a, methyl ester of
3,4-secoacid 23a and dimethyl esters of 2,3-secodiacids 43a and 43b. However, we
obtained the corresponding orthoacetates (15a, 26a, 45a and 45b), the yields of the
desired glucoside tetraacetates being very low. The orthoacetates are unstable and de-
compose during purification or spectral measurements in chloroform under formation
of mixtures containing (according to thin-layer chromatography) the starting aglycones
and two tetra-O-acetyl derivatives of glucose. In the case of orthoacetate 26a the de-
1
composition products were identified by H and 13C NMR spectra as the aglycone 23a,
1,3,4,6-tetra-O-acetyl-α-D-glucopyranose and 2,3,4,6-tetra-O-acetyl-α-D-glucopyranose
(the NMR data of both glucose tetraacetates agree well with the published ones21,22).
Much more successful for the preparation of glucoside tetraacetates proved to be the
presence of a soluble catalyst - mercury(II) cyanide in acetonitrile. This method was
already used for the synthesis of triterpenoid glycosides, e.g. lupane10 and dammarane23
derivatives. Using mercury(II) cyanide we obtained the 28-O-glucoside tetraacetates
16b, 20a, 20b, 27a, 27b, 46a and 46b, and 3β-O-glucoside acetates 4a and 4b. In
addition to the glucoside acetates, we often isolated from the reaction mixtures also the
unreacted starting aglycones and in some cases their acetates; the formation of ortho-
acetates was not observed. From 3,4-seco-3,28-diol 32a we obtained predominantly the
bis-glucoside tetraacetate 36a along with 3-O-glucoside tetraacetate 38a; minor
amounts of the isomeric glucoside tetraacetates 40a and 41a with acetylated hydroxyl
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

1778
Klinotova et al.:
in the aglycone part (in position 3 and 28, respectively) were also found. These acetates
were obtained as an unseparable 1 : 1 mixture (according to NMR spectra) and were
1
identified by H and 13C NMR spectra of the mixture.
1
1
R
R
2
2
COOR
CH OR
2
3
R O
X
3
2
2
R
X
R
H
R
H
H
H
O
1a, 1b
12a, 12b
13a, 13b
14a, 14b
15a
CH
H
Ac
Ac
O
2a, 2b
3a, 3b
4a, 4b
5a, 5b
6a, 6b
7a, 7b
8a, 8b
9a, 9b
10a
3
Ac
NOH
O
CH
CH
Glc(Ac)
Glc
Glc-orthoAc
3
3
4
16a, 16b Glc(Ac)
O
4
Glc-orthoAc Ac
Glc
O
17a, 17b
Glc(Ac)
Glc
Ac
4
Ac
Glc
H
Glc(Ac)
Glc
Glc(Ac)
Glc
4
4
11a
1
1
R
R
2
2
CH OR
CH OR
2
2
3
NC
R OOC
3
2
2
R
R
R
H
H
H
H
18a, 18b
19a, 19b
20a, 20b
21a, 22b
22a, 22b
Ac
CH
3
23a, 23b
24a, 24b
25a, 25b
26a
Glc(Ac)
Glc
Ac
Ac
H
4
CH
3
Glc-orthoAc CH
3
3
Glc(Ac)
CH
CH
27a, 27b
28a, 28b
29a, 29b
30a
4
Glc
Glc
Ac
3
H
Glc(Ac)
4
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

Triterpenes
1779
The same glycosylation protocol was also applied to the preparation of derivatives 7a
and 7b in which the glucose moiety is bonded by ester bond to the carboxyl group.
Glycosylation of acetylbetulinic acid (3a) and acetyldihydrobetulinic acid (3b) af-
forded, beside glucosyl acetates 7a and 7b, also orthoacetates 6a and 6b. They were
1
identified by H NMR spectra as an impurity in the crude reaction product. The men-
tioned glycosylation method proved to be unsuitable for the preparation of diglucosyl
derivatives of betulinic acid (1a) and dihydrobetulinic acid (1b) because these acids are
1
1
R
R
2
2
CH OR
CH OR
2
2
3
R OOC
3
R OOC
3
R O
3
3
2
2
R
R
H
R
R
H
H
H
H
42a, 42b
32a
33a
34a
35a
36a
37a
38a
39a
40a
41a
CH
CH
43a, 43b
44a, 44b
45a, 45b
46a, 46b
47a, 47b
Ac
H
Ac
Ac
H
3
3
3
3
3
Ac
Glc-othoAc CH
Ac
Glc(Ac)
Glc
CH
CH
Glc(Ac)
Glc
Glc(Ac)
Glc
4
4
4
4
4
4
H
Glc(Ac)
Glc
H
Ac
Glc(Ac)
Ac
Glc(Ac)
1
a, R = C(CH )=CH
In formulae 1-47 :
3
2
1
b, R = CH(CH )
3 2
CH OH
2
CH OAc
2
O
O
O
O
OH
OAc
Glc =
HO
Glc(Ac) =
4
AcO
OH
OAc
CH OAc
2
O
OAc
Glc-orthoAc =
Ac = COCH
3
AcO
O
O
O
CH
3
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

1780
Klinotova et al.:
very sparingly soluble in acetonitrile or its mixtures with ether and chloroform.
Dihydrobetulinic acid (1b) was recovered entirely unchanged from the reaction mix-
ture, betulinic acid (1a) afforded small amounts of bis-glucosyl derivative 10a.
Glucosides 5a, 5b, 8a, 8b, 11a, 17a, 17b, 21a, 21b, 28a, 28b, 37a, 39a, 47a and 47b
were obtained from the corresponding glucoside acetates by treatment with sodium
methoxide in methanol at room temperature. Under these conditions only the acetate
groups in the sugar part of the molecule were cleaved off whereas the ester groups in
the aglycone part remained intact. The acetate group in position 3 of glucosides 8a and
8b was hydrolyzed with sodium hydroxide in methanol at 50 °C under formation of
glucosides 9a and 9b. Analogous hydrolysis of the methyl ester group in the 3,4-seco
derivatives 28a and 28b proceeded already at room temperature and afforded gluco-
sides 29a and 29b; under same conditions the methyl ester groups in the 2,3-seco deri-
vatives 47a and 47b were not hydrolyzed. Glucosides 9a and 11a, derived from
betulinic acid (1a), have already been prepared by other methods24–26
.
Structure of the prepared compounds has been confirmed by infrared, NMR and
mass spectra (electron impact spectra (EI-MS) in the case of aglycones and glucoside
acetates, fast atom bombardment spectra (FAB-MS) in the case of glucosides). Electron
impact spectra exhibited characteristic very abundant ions of m/z 331, 169 and 109,
arising from the glucose units, in the higher mass region the fragmentation of these
compounds corresponds to the known13,15,27 fragmentation of the corresponding agly-
cone. In addition to ions [M + Na]+ and [M + H]+, FAB mass spectra of glucosides
displayed fragment ions [M + H – 162]+ and [M + H – 180]+ corresponding to loss of
glucosyl or glucosyloxyl group. Moreover, compounds with glucose esterically bonded
to the carboxyl group in position 17 typically undergo cleavage of the C(17)-C(28)
bond leading to ions [M + H – 208]+. No meaningful FAB spectra were obtained in the
1
case of diglucoside 37a; however, its structure was confirmed by the H NMR spec-
trum.
Proton NMR data for all the glucoside acetates prepared are summarized in Tables I–III,
spectra of some glucosides (5a, 8a, 11a, 29a, 37a, 39a) are given in Table IV. The
values of the coupling constants J(1′,2′) (about 8 Hz) show that all these derivatives
have β-configuration at the anomeric carbon atom. In the case of orthoacetates 26a and
45a, the chemical shifts and coupling constants of protons in the glucose moiety
(Tables II and III) agree with the data for analogous orthoacetates of the α-D-gluco-
configuration28,29; the chemical shift of the singlet of the methyl group on the dioxolane
ring (δ about 1.73) corresponds to the endo-configuration of this methyl group29.
The assignment of chemical shifts in the 13C NMR spectra of aglycones and gluco-
side acetates (Tables V and VI) is based on the APT spectra and comparison with the
literature data19,30,31 for aglycones 2a, 12a, 32a and 33a and for similar lupane deriva-
tives and their glucoside acetates10,11. Attachement of tetra-O-acetyl-β-D-glucopyrano-
syl group to the 28-hydroxyl group results in a large downfield shift of the C-28 signal
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

Triterpenes
1781
TABLE I
Proton NMR data of compounds 4a, 4b, 7a, 7b, 10a, 16a and 16b in CDCl3
Chemical shifts (coupling constants)
Proton
4a
4b
7a
7b
10a
16a
16ba
Aglycone moiety
0.831 s
CH3 0.714 s
0.809 s
0.719 s
0.819 s
0.893 s
0.893 s
0.826 s
0.833 s
0.841 s
0.886 s
0.711 s
0.809 s
0.876 s
0.881 s
0.936 s
0.982 s
1.036 s
1.070 s
1.078 s
2.40 ddd
15.8;7.6;4.4
2.50 ddd
15.8;9.7;7.6
–
0.948 bs
0.960 bs
1.041 s
0.839 s
0.887 s
0.900 s
0.850 s
0.883 s
1.076 s
0.947 s
0.927 s
0.948 s
0.929 s
0.939 s
1.083 s
b
b
b
b
b
H-2α
2.41 ddd
15.7;7.5;4.4
2.50 ddd
15.7;9.8;7.5
–
b
b
b
b
b
H-2β
H-3
3.05 dd
3.07 dd
4.46 dd
~11.0;5.8
2.94 td
4.47 dd
3.06 dd
11.8;4.8
2.94 td
11.5;4.5
H-19 2.99 td
10.8;10.8;4.6
11.4; 4.8
11.4;5.5
b
b
b
2.37 td
~11.1;11.1;4.6
–
~11.0;11.0;4.7
–
~10.8;10.8;5.8
3.54 dd
9.2;1.3
3.64 bd
9.2
H-28a
–
–
–
–
–
3.52 dd
9.4;1.3
3.61 bd
9.4
H-28b
–
–
–
H-29a 4.60 m
4.61 m
4.61 m
4.58 m
0.745 d
0.744 d
0.752 d
H-29b 4.72 bd
~2.4
7.0
4.74 bd
~2.2
6.7
4.74 bd
~2.4
4.67 bd
~2.4
~6.7
H-30 1.685 bs
0.850 d
7.0
1.678 bs
0.847 d
1.678 bs
1.670 bs
0.838 d
6.7
~6.7
OMe 3.67 s
3.65 s
–
–
–
–
–
–
–
–
–
OAc
–
2.041 s
2.045 s
Glucose moiety
10ac
10ad
5.70 d
8.3
H-1
H-2
H-3
H-4
H-5
4.53 d
4.53 d
7.8
5.70 d
8.3
5.68 d
4.53 d
4.47 d
7.8
4.46 d
7.8
8.3
8.0
8.0
5.03 dd
9.6;7.8
5.20 t
5.03 dd
9.6;7.8
5.20 t
5.21 dd
9.5;8.3
5.28 t
9.5;9.3
5.16 dd
5.20 dd
9.6;8.3
5.27 dd
9.6;9.2
5.16 dd
10.1;9.2
3.82 ddd
5.02 dd
9.7;8.0
5.20 t
5.21 dd
9.7;8.3
5.28 t
5.02 dd
9.6;7.8
5.23 t
5.01 dd
9.7;8.0
5.22 dd
9.7;9.4
5.10 dd
9.9;9.4
3.70 ddd
9.9;4.6;2.6
4.14 dd
12.2;2.6
4.29 dd
12.2;4.6
2.012 s
2.019 s
2.028 s
2.097 s
~9.5;9.5
5.04 dd
~10.0;9.5
3.68 ddd
~9.5;9.5
5.05 dd
~9.5;9.5
5.04 dd
~10.0;9.5
3.68 ddd
~9.4;9.4
5.16 dd
9.8;9.3
3.83 ddd
~9.6;9.6
5.11 t
~10.0;9.5 ~10.0;9.3
3.68 ddd 3.83 ddd
~9.7;9.7
3.70 ddd
10.0;5.5;2.5 10.0;5.5;2.5 10.0;4.4;2.4
10.1;4.4;2.4 10.0;5.4;2.5
10.2;4.4;2.4 10.0;4.6;2.4
H-6b 4.11 dd
12.2;2.5
H-6b 4.25 dd
12.2;5.5
4.11 dd
12.2;2.5
4.25 dd
12.2;5.5
2.007 s
2.025 s
2.031 s
2.077 s
4.07 dd
12.4;2.4
4.32 dd
12.4;4.4
2.020 s
2.024 s
2.037 s
2.078 s
4.06 dd
12.4;2.4
4.31 dd
12.4;4.4
2.014 s
2.019 s
2.034 s
2.075 s
4.11 dd
12.3;2.5
4.24 dd
12.3;5.5
2.005 s
2.022 s
2.027 s
2.073 s
4.07 dd
12.5;2.4
4.32 dd
12.5;4.4
2.017 s
2.023 s
2.036 s
2.075 s
4.15 dd
12.0;2.4
4.30 dd
12.0;4.6
2.016 s
2.024 s
2.031 s
2.102 s
OAc 2.005 s
2.023 s
2.029 s
2.072 s
a
b
c
H-1β: 1.91 ddd (J = 13.3; 7.5 and 4.4 Hz); signal was not assigned; 3-O-Glc(Ac)4; d 28-O-Glc(Ac)4.
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

1782
Klinotova et al.:
TABLE II
Proton NMR data of compounds 18a, 18b, 20a, 20b, 23a, 26a, 27a, 27b and 30a in CDCl3
Chemical shifts (coupling constants)
Proton
18a
18b
20a
20b
23a
26aa
27a
27b
30a
Aglycone moiety
CH3 0.838 s
0.993 bs
0.850 s 0.846 s
0.976 bs 0.981 s
1.078 s 1.078 s
0.857 s
0.966 s
1.086 s
0.827 s
0.992 s
1.066 s
0.830 s
0.978 s
1.075 s
0.834 s
0.981 s
1.073 s
0.845 s
0.963 s
1.080 s
0.818 s
0.981 s
1.068 s
1.071 s
H-2a 2.17 m
H-2b 2.31 m
2.19 m 2.16 m
2.32 m 2.31 m
2.19 m
2.15 m
2.33 m
2.39 td
2.15 m
2.33 m
2.37 td
2.15 m
2.33 m
2.36 td
2.17 m
2.17 m
2.37 m
2.32 m
b
2.34 m
b
b
H-19 2.39 td
2.37 td
2.45 td
~11;11;5.8
~11;11;5.7
4.64 bd 4.65 bd
~2.0 ~2.0
~11;11;5.8 ~10.8;10.8;5.5 ~11;11;5.7
~11;11;5.7
H-23a 4.64 bd
4.65 bd
~2.0
4.65 bd
4.65 bd
4.66 bd
4.66 bd
~2.1
4.64 bd
~2.2
~2.2
~2.0
~2.3
~2.1
H-23b 4.88 m
H-24 1.719 m
4.88 m 4.89 m
1.723 m 1.729 m
4.89 m
4.85 m
4.85 m
4.85 m
4.86 m
4.84 m
1.733 m
1.723 m
1.725 bs
1.732 m
1.736 m
1.708 m
H-28a 3.35 dd
3.32 dd 3.53 dd
3.51 dd
3.34 bd
3.17 d
3.54 dd
3.52 dd
3.86 bd
10.8;1.1
10.9;1.2 9.3;1.5
9.4;1.4
11.0
8.8
9.2;1.6
9.3;1.5
11.2
H-28b 3.79 dd
3.76 dd 3.64 bd
3.61 bd
3.80 bd
3.59 dd
3.63 bd
3.61 bd
4.25 dd
10.8;1.9
10.9;1.7 9.3
9.4
11.0
8.8;1.2
9.2
9.3
11.2;1.8
H-29 4.61 m
4.59 m
0.778 d
4.59 m
4.59 m
4.58 m
4.61 m
0.759 d
0.756 d
H-29b 4.69 bd
~2.4
6.5
4.68 bd
~2.4
7.0
4.69 bd
~2.4
4.68 bd
~2.3
4.67 bd
~2.4
6.5
4.70 bd
~2.3
H-30 1.690 m
0.850 d 1.674 m
0.840 d
1.688 bs
1.679 bs
1.672 m
0.836 d
1.698 m
6.5
7.0
6.5
OMe
OAc
–
–
–
–
–
–
–
–
3.65 s
–
3.65 s
–
3.65 s
–
3.66 s
–
–
2.074 s
Glucose moiety
H-1
–
–
–
–
–
–
–
–
–
–
–
–
–
–
4.46 d
8.0
4.45 d
8.0
–
–
–
–
–
–
–
5.69 d
5.2
4.46 d
8.0
4.45 d
8.0
5.71 d
8.2
H-2
5.02 dd
9.6;8.0
5.01 dd
9.7;8.0
4.31 dd
5.2;3.1
5.02 dd
9.7;8.0
5.01 dd
9.7;8.0
5.13 dd
9.6;8.2
H-3
5.22 t
~9.7;9.7
5.22 t
~9.7;9.7
5.19 t
3.0;3.0
5.22 dd
9.7;9.4
5.22 dd
9.7;9.4
5.24 t
9.4;9.4
H-4
5.10 t
~9.7;9.7
5.10 t
~9.6;9.6
4.91 dd
9.6;2.9
5.10 dd
9.9;9.4
3.70 ddd
5.10 dd
10.0;9.4
3.69 ddd
5.13 t
9.4;9.4
H-5
3.70 ddd
9.8;4.6;2.6 9.9;4.6;2.6
3.69 ddd
3.97 ddd
9.4;5.2;3.0
3.83 ddd
9.9;4.6;2.6 10.0;4.6;2.6 9.8;4.5;2.3
H-6b
H-6b
OAc
4.15 dd
12.4;2.6
4.14 dd
12.4;2.6
4.19 dd
12.2;3.0
4.15 dd
12.2;2.6
4.14 dd
12.2;2.6
4.10 dd
12.3;2.3
4.30 dd
12.4;4.6
4.29 dd
12.4;4.6
4.22 dd
12.2;5.2
4.30 dd
12.2;4.6
4.29 dd
12.2;4.6
4.29 dd
12.3;4.5
2.014 s
2.017 s
2.030 s
2.101 s
b
2.011 s
2.014 s
2.027 s
2.097 s
2.098 s
2.105 s
2.121 s
2.013 s
2.018 s
2.029 s
2.101 s
2.010 s
2.015 s
2.026 s
2.096 s
2.006 s
2.017 s
2.034 s
2.087 s
–
–
–
a
Orthoacetate methyl group: 1.74 s; signal was not assigned.
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

Triterpenes
1783
TABLE III
Proton NMR data of compounds 36a, 38a, 40a, 41a, 45a, 46a and 46b in CDCl3
Chemical shifts (coupling constants)
40aa 41aa
Aglycone moiety
Proton
36a
38a
45ab
46a
46b
CH3
0.782 s
0.976 s
0.774 s
0.987 s
0.774 s
0.982 s
0.804 s
0.991 s
0.910 s
1.000 s
1.034 s
2.25 d
~18.0
2.42 d
~18.0
–
0.907 s
1.000 s
1.031 s
2.26 d
18.1
2.43 d
18.1
–
0.922 s
0.978 s
1.038 s
2.29 d
17.8
2.44 d
17.8
–
1.062 s
1.054 s
1.061 s
1.074 s
c
c
c
c
H-1a
H-1b
H-3a
H-3b
H-19
H-23a
H-23b
c
c
c
c
3.34 dt
3.34 dt
3.34 dt
3.98 m
3.98 m
2.37 td
9.2;7.1;7.1
3.85 ddd
9.2;6.7;5.5
2.36 td
11.0;11.0;5.8
4.60 bd
9.2;7.1;7.1
3.85 ddd
9.2;6.8;5.4
2.39 td
10.8;10.8;5.8 11.0;11.0;5.7 11.0;11.0;5.7
4.60 bd
9.3;7.0;7.0
3.85 ddd
9.2;6.7;5.5
2.45 td
–
–
–
c
c
2.36 td
10.8;10.8;5.8
4.59 bd
~2.2
4.81 m
4.64 bd
~2.2
4.84 m
~2.2
4.82 m
~2.0
4.81 m
1.225 s
1.231 s
1.242 s
H-24
H-28a
1.703 bs
3.54 bd
9.4
3.63 bd
9.4
1.692 bs
3.34 bd
11.0
3.80 dd
11.0;1.6
4.60 m
1.692 m
3.86 bd
11.0
4.24 dd
11.0;1.5
4.61 m
1.722 m
3.54 bd
9.3
3.63 bd
9.3
1.233 m
3.17 bd
~9.0
3.58 bd
~9.0
1.239 s
3.53 bdd
9.2;1.3
3.63 bd
9.2
1.247 s
3.51 bd
9.2
3.60 bd
9.2
H-28b
H-29a
H-29b
H-30
4.59 m
4.59 m
4.57 m
4.56 m
0.744 d
~7.0
4.67 bd
~2.4
1.675 bs
4.69 bd
~2.4
1.692 bs
4.70 bd
~2.4
1.692 m
4.68 bd
~2.4
1.682 bs
4.67 bd
~2.2
1.66 bs
4.66 bd
~2.4
1.658 m
0.824 d
~7.0
OMe
–
–
–
–
–
–
3.61 s
3.64 s
–
3.61 s
3.64 s
–
3.62 s
3.65 s
–
OAc
2.073 s
2.041 s
Glucose moiety
36ad
4.47 d
8.0
36ae
4.46 d
8.0
H-1
H-2
H-3
H-4
H-5
4.47 d
8.0
4.48 d
8.0
4.47 d
8.0
5.69 d
5.2
4.31 dd
5.2;3.0
5.18 t
3.0;3.0
4.91 dd
9.5;2.8
3.96 ddd
4.45 d
8.0
4.44 d
8.0
5.00 dd
9.7;8.0
5.21 t
4.98 dd
9.7;8.0
5.19 t
9.5;9.5
5.08 t
9.8;9.8
5.02 dd
9.7;8.0
5.22 t
9.6;9.6
5.10 t
9.8;9.8
4.98 dd
9.7;8.0
5.19 dd
9.7;9.5
5.08 dd
9.9;9.5
3.68 ddd
4.98 dd
9.7;8.0
5.19 t
9.5;9.5
5.08 dd
9.9;9.5
3.68 ddd
5.02 dd
9.7;8.0
5.22 t
9.6;9.6
5.11 t
5.01 dd
9.7;8.0
5.22 dd
9.7;9.4
5.10 dd
9.9;9.4
3.70 ddd
9.7;9.4
5.10 dd
9.9;9.4
3.69 ddd
9.9;4.6;2.6
4.14 dd
12.2;2.6
4.29 dd
12.2;4.6
2.009 s
2.012 s
2.025 s
2.093 s
9.7;9.7
3.70 ddd
3.68 ddd
3.70 ddd
10.0;4.6;2.5 10.0;4.6;2.5 9.9;4.6;2.4
10.0;4.6;2.4 10.0;4.6;2.4 ~10.0;5.0;3.0 9.9;4.6;2.6
H-6a 4.13 dd
12.4;2.5
H-6b 4.26 dd
12.4;4.6
OAc 2.006 s
2.024 s
4.15 dd
12.4;2.5
4.30 dd
12.4;4.6
2.013 s
2.018 s
2.029 s
2.103 s
4.13 dd
12.4;2.4
4.26 dd
12.4;4.6
2.005 s
2.024 s
2.026 s
2.085 s
4.13 dd
12.4;2.6
4.26 dd
12.4;4.6
2.007 s
2.025 s
2.028 s
2.086 s
4.15 dd
12.4;2.6
4.30 dd
12.4;4.6
2.014 s
2.020 s
2.030 s
2.103 s
b
4.19 m
4.22 m
4.14 dd
12.2;2.6
4.29 dd
12.2;4.6
2.012 s
2.016 s
2.027 s
2.098 s
2.095 s
2.101 s
2.119 s
2.026 s
2.086 s
a
c
Data were obtained from the mixture of compounds 40a and 41a; orthoacetate methyl group: 1.73 s; signal was not
assigned; d 3-O-Glc(Ac)4; 28-O-Glc(Ac)4.
e
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

1784
Klinotova et al.:
TABLE IV
Proton NMR data of compounds 5a, 8a, 11a, 29a, 37a and 39a in CD3OD
Chemical shifts (coupling constants)
Proton
5a
8a
11a
29a
Aglycone moiety
37a
39a
CH3 0.826 s
0.868 s
0.852 s
0.861 s
0.893 s
0.967 s
1.014 s
4.44 dd
11.2;5.2
–
0.824 s
0.865 s
0.957 s
1.000 s
1.030 s
3.15 dd
11.4;4.4
–
0.859 s
1.053 s
1.137 s
1.747 bs
0.824 s
1.045 s
1.131 s
1.729 bs
0.823 s
1.045 s
1.122 s
1.728 bs
0.935 s
1.003 s
1.033 s
H-3a 3.15 dd
11.5;4.5
–
–
3.40 td
8.6;8.6;6.4
3.84 td
8.6;8.6;5.4
2.47 td
10.5;10.5;5.6
4.66 bd
2.4
3.40 td
8.6;8.6;6.4
3.84 td
9.0;9.0;5.5
2.43 td
10.6;10.6;5.6
4.66 bd
2.4
H-3b
–
H-19 2.99 td
3.01 td
3.00 td
10.7;10.7;4.6
a
2.47 td
10.8;10.8;5.5
4.68 bd
2.4
10.8;10.8;5.0 11.0;11.0;5.0
a
a
H-23a
a
a
a
H-23a
H-28a
4.85 m
3.63 dd
9.5;1.5
3.75 bd
9.5
4.82 m
3.63 bd
9.6
4.82 m
3.29 bd
11.2
–
–
–
H-28b
–
–
–
3.75 bd
9.6
3.75 dd
11.2;1.6
4.59 m
4.70 bd
2.4
H-29a 4.60 m
H-29b 4.72 bd
2.2
4.60 m
4.72 bd
2.2
4.60 m
4.72 bd
2.2
4.59 m
4.70 bd
2.4
4.58 m
4.70 bd
2.5
H-30 1.694 bs
OMe 3.66 s
1.698 bs
–
1.695 bs
–
1.707 bs
–
1.703 bs
–
1.702 bs
–
OAc
–
2.02 s
–
–
–
–
Glucose moiety
11ab
11ac
37ab
4.24 d
7.8
37ac
4.23 d
7.8
H-1
H-2
H-3
H-4
H-5
4.31 d
7.8
5.49 d
8.1
4.31 d
7.8
5.50 d
8.2
4.23 d
7.8
4.24 d
7.8
3.18 dd
8.8;7.8
3.33 t
8.6;8.6
3.28 t
9.2;9.2
3.23 ddd
9.4;5.4;2.2
3.32 dd
8.8;8.2
3.18 dd
8.8;7.8
3.32 t
8.2;8.2
3.19 dd
7.8;9.0
3.17 dd
9.0;7.8
3.34 t
8.5;8.5
3.30 t
9.0;9.0
3.24 ddd
9.0;5.5;2.4
3.68 dd
11.8;5.4
3.85 dd
11.8;2.4
3.19 dd
9.0;7.8
3.36 t
9.0;9.0
~3.29 m
3.17 dd
9.0;8.0
3.34 t
8.6;8.6
3.30 t
8.6;8.6
3.24 ddd
9.0;5.5;2.4
3.68 dd
11.8;5.5
3.85 dd
11.8;2.4
3.37–3.45 m 3.33 t
8.8;8.8
3.37–3.45 m 3.36 t
9.0;9.0
3.37–3.45 m 3.28 t
8.8;8.8
3.37–3.45 m ~3.28 m
3.37–3.45 m 3.24 ddd
9.4;5.4;2.4
3.37–3.45 m ~3.28 m
~3.29 m
3.69 m
H-6a 3.66 dd
11.7;5.4
3.70 m
3.66 dd
11.6;5.4
3.835 dd
11.6;2.4
3.70 m
3.69 m
H-6b 3.84 dd
11.7;2.2
3.84 dd
11.8;1.6
3.84 dd
11.8;1.5
3.89 dd
11.8;2.0
3.89 dd
11.6;1.8
a
c
Signal is presented in methyl groups; b 3-O-Glc; 28-O-Glc.
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

Triterpenes
1785
TABLE V
Carbon-13 chemical shifts of compounds 4a, 4b, 7a, 7b, 10a, 16b, 18a, 18b, 20a and 20b in CDCl3
Chemical shifts (coupling constants)
Carbon
4a
4b
7a
7b
10a
16b
18a
18b
20a
20b
Aglycone moiety
38.59
25.86
90.65
38.93
55.57
18.13
34.35
40.63
50.48
36.87
20.84
25.45
38.11
42.37
29.64
31.66
56.78
49.11
46.70
150.19
30.34
36.44
27.56
16.11
16.00
15.95
14.60
174.15
109.77
19.39
–
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
OAc
38.64
25.89
90.74
38.95
55.65
18.14
34.28
40.67
50.53
36.89
20.86
25.50
38.22
42.35
29.62
32.15
56.55
49.47
46.93
150.64
30.61
36.95
27.59
16.13
16.00
15.91
14.61
176.66
109.55
19.39
–
38.64
25.89
90.70
38.96
55.61
18.14
34.36
40.68
50.30
36.87
20.89
26.90
38.08
42.49
29.63
32.07
56.98
48.94
44.17
38.32
23.63
80.85
37.74
55.34
18.13
34.28
40.62
50.38
37.05
20.83
25.38
38.13
42.38
29.64
31.66
56.77
49.07
46.75
38.34
23.63
80.86
37.75
55.32
18.14
34.37
40.67
50.16
37.03
20.87
26.79
38.00
42.54
29.60
31.59
57.19
48.57
44.07
29.70
22.54
36.86
27.89
16.45
16.10
15.96
14.52
174.51
22.84
14.60
171.05
21.29
–
39.50
34.09
218.05
47.31
54.83
19.58
33.55
40.83
49.34
36.79
21.29
26.76
37.30
42.92
26.89
29.47
47.19
47.88
44.66
29.47
21.56
34.68
26.58
21.04
15.84
15.84
14.62
68.64
22.90
14.81
–
32.50
11.39
120.22
146.99
50.47
24.18
34.41
40.60
40.41
39.39
21.46
24.81
37.23
43.07
26.95
28.99
47.71
48.56
47.72
150.23
29.62
33.88
113.99
22.74
19.74
15.92
14.56
60.44
109.93
19.00
–
32.58
11.37
120.24
147.01
50.45
24.17
34.34
40.66
40.06
39.40
21.49
26.47
36.72
43.25
26.83
29.16
47.85
47.92
44.47
29.46
21.67
33.95
114.00
22.75
19.68
15.95
14.43
60.51
22.89
14.84
–
32.50
11.40
120.17 120.18
146.92 146.93
32.59
11.37
50.46
24.16
34.41
40.58
40.43
39.40
21.46
24.76
37.60
43.08
26.96
29.30
46.99
48.46
47.89
150.16
29.53
34.59
50.44
24.15
34.34
40.62
40.07
39.40
21.50
26.40
37.15
43.27
26.86
29.42
47.17
47.82
44.63
29.47
21.56
34.66
29.72 150.12
22.73
37.30
27.59
16.13
15.97a
15.93a
14.53b
30.31
36.44
27.87
16.43
16.13
15.94
14.62
114.05 114.06
22.78
19.72
15.99
14.67
68.49
109.90
18.97
–
22.79
19.65
16.02
14.53
68.60
22.89
14.82
–
176.88 174.22
22.94 109.83
14.67b
–
19.30
171.05
21.28
–
OMe
51.24
51.14
–
–
–
–
–
–
Glucose moiety
10ac
102.95
71.48
72.84
68.74
71.62
62.25
170.68
170.42
169.45
169.19
20.73
20.66
20.61
20.59
10ad
91.04 101.64
1
2
3
4
5
6
OAc
102.96
71.50
72.87
68.79
71.66
62.28
170.67
170.40
169.44
169.18
20.73
20.67
20.60
20.58
102.97
71.50
72.87
68.76
71.65
62.24
170.68 170.62
170.40 170.19
169.44 169.42
169.18 168.97
91.02
69.86
72.49a
67.95
72.85a
61.47
90.99
69.86
72.46a
67.97
72.92a
61.50
170.62
170.19
169.42
168.95
20.65
20.55
20.55
20.52
–
–
–
–
–
–
–
–
–
–
–
–
–
–
101.55 101.61
69.88
72.51a
67.96
72.84a
61.47
170.68 170.73
170.17 170.41
169.43 169.42
169.00 169.20
20.69
20.55
20.55
20.55
c
71.27
72.77
68.59
71.68
62.05
71.22
72.73
68.54
71.71
62.00
71.25
72.74
68.57
71.67
62.04
170.72 170.72
170.40 170.41
169.42 169.42
169.18 169.18
20.73
20.68
20.60
20.58
20.66
20.55
20.52
20.52
20.73
20.61
20.58
20.58
–
–
20.73
20.60
20.57
20.57
20.73
20.61
20.57
20.57
a,b
d
Signals with identical symbols may be mutually interchanged; 3-O-Glc(Ac)4; 28-O-Glc(Ac)4.
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

1786
Klinotova et al.:
TABLE VI
Carbon-13 chemical shifts of compounds 23a, 27a, 27b, 30a, 36a, 38a, 40a, 41a, 46a and 46b in CDCl3
Chemical shifts (coupling constants)
Carbon
23a
27a
27b
30a
36a
38a
40aa
41aa
46a
46b
Aglycone moiety
32.71
23.09
71.21
148.04
50.24
24.54
35.47
40.50
40.58
39.09
21.25
25.06
37.73
43.07
27.05
29.31
47.04
48.56
47.90
150.33
29.59
34.60
113.04
23.23
20.40
16.02
14.77
68.60
109.79
19.02
–
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
OAc
32.69
28.38
174.62
147.56
50.25
24.53
34.06
40.59
40.62
39.19
21.33
25.10
37.35
43.09
27.03
29.06
47.75
48.67
47.73
150.37
29.69
33.91
113.39
23.24
20.04
15.94
14.60
60.50
109.80
19.06
–
32.68
28.35
174.55 174.58
147.49 147.50
32.76
28.36
32.57
28.58
172.14
147.34
50.27
24.39
33.82
40.54
40.63
39.17
21.29
25.05
37.58
43.07
27.03
29.63b
46.28
48.66
47.65
150.03
29.54b
34.50
113.67
23.10
20.00
15.95
14.63
62.73
110.03
19.11
171.65
21.02
–
32.68
23.08
71.24
148.09
50.25
24.54
35.46
40.52
40.55
39.08
21.23
25.11
37.34
43.05
27.01
29.04
47.74
48.66
47.72
150.38
29.68
33.90
112.97
23.17
20.41
15.93
14.64
60.47
109.79
19.04
–
32.68
23.11
71.25
148.11
50.29
24.53
35.45
40.52
40.56
39.08
21.22
25.13
37.63
43.08
27.06
29.45
46.28
48.70
47.65
150.09
29.65
34.50
112.98
23.17
20.41
16.02
14.67
62.77
109.96
19.10
171.67
21.03
–
32.68
22.15
65.05
147.89
50.29
24.53
35.29
40.51
40.56
39.12
21.38
25.04
37.76
43.08
27.06
29.32
47.06
48.57
47.92
150.42
29.60
34.60
113.14
23.09
20.36
16.02
14.78
68.58
109.71
19.10
171.20
20.98
–
41.91
171.93 172.00
179.98 179.97
41.94
46.29
48.30b
20.63
33.13
40.73
41.77b
41.52
21.78
25.23
37.83
43.04
27.04
29.26
47.08
48.49
47.89
150.38
29.58
34.56
27.81
23.67
19.68
15.93
14.81
68.63
109.69
18.97
–
46.36
48.32b
20.70
33.25
40.78
41.63b
41.63
21.81
26.93
37.37
43.24
26.93
29.38
47.24
47.83
44.59
29.42
21.54
34.63
27.70
23.77
19.64
15.98
14.84
68.71
22.89
14.67
–
50.22
24.51
34.04
40.55
40.61
39.19
21.32
25.02
37.70
43.08
27.04
29.32
47.03
48.54
47.88
150.30
29.58
34.60
50.18
24.48
33.99
40.57
40.24
39.15
21.32
26.69
37.24
43.27
26.90
29.43
47.18
47.86
44.63
29.46
21.54
34.66
113.44 113.45
23.28
20.03
16.00
14.70
68.56
109.77
19.02
–
23.30
19.96
16.03
14.56
68.69
22.89
14.81
–
OMe
51.55
51.56
51.58
–
–
50.79
51.72
50.85
51.72
Glucose moiety
36ad
101.20
71.27
72.85
68.42
71.77
61.97
170.75
170.37
169.42
169.22
20.74
20.61
20.61
20.59
36ae
101.63 101.19
1
2
3
4
5
6
OAc
–
–
–
–
–
–
–
101.60 101.67
91.61
70.21
72.76c
67.81
72.71c
61.47
170.62
170.13
169.40
169.12
20.67
20.55
20.55
20.45
101.22
71.27
72.86
68.43
71.77
61.97
170.73
170.37
169.43
169.22
20.72
20.61
20.61
20.59
101.62
71.24
72.76
68.55
71.71
62.02
170.73
170.37
169.43
169.22
20.72
20.61
20.59
20.59
101.63 101.68
71.24
72.76
68.57
71.72
62.02
71.26
72.75
68.56
71.66
62.03
71.24
72.76
68.55
71.71
62.02
170.75 170.72
170.40 170.36
169.42 169.42
169.22 169.22
20.72
20.61
20.59
20.59
71.26
72.84
68.41
71.75
61.95
71.25
72.75
68.56
71.70
62.02
71.27
72.77
68.59
71.66
62.04
170.72 170.75
170.39 170.41
169.42 169.43
169.19 169.22
170.72 170.72
170.39 170.40
169.42 169.42
169.19 169.20
–
20.73
20.61
20.58
20.58
20.73
20.62
20.58
20.58
20.70
20.59
20.59
20.56
b,c
20.73
20.60
20.57
20.57
20.73
20.61
20.57
20.57
a
Data have been obtained from the mixture of compounds 40a and 41a;
signals with identical symbols may be mu-
d
e
tually interchanged; 3-O-Glc(Ac)4; 28-O-Glc(Ac)4.
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

Triterpenes
1787
(+8.1 ppm) and a small upfield shift of the C-17 signal (–0.7 ppm). Smaller shifts were
observed for the more distant carbon atoms (C-13: +0.4 ppm; C-16: +0.3 ppm; C-22:
+0.7 ppm). Glucosylation of the 3β-OH group results in downfield shift of C-3 (+12.1 ppm)
and upfield shift of C-2 (–1.4 ppm) signals; the signal of quaternary C-4 atom is in-
fluenced only a little (+0.2), less than signals of C-5 (+0.4 ppm) and of carbon atoms in
methyl groups attached in position 4 (C-23: –0.3 ppm; C-24: +0.8). In the 13C NMR
spectra of 3-hydroxy-3,4-seco derivatives there are significant shifts of the C-3 (+7.9 ppm)
and C-2 (–2.9 ppm) signals and also signals of the more distant atoms C-6 (–0.4 ppm)
and C-7 (+0.4 ppm).
Small but systematic differences occur in the chemical shifts of protons and carbon
atoms of the glucose part in glucoside acetates with the sugar moiety attached in posi-
1
tion 3 or 28. On this basis we assigned the H and 13C NMR signals for diglucoside
acetates 10a and 36a to the individual glucose moieties in positions 3 and 28. The same
principle was also applied to the isomeric glucoside acetates 40a and 41a (obtained
only as a 1 : 1 mixture).
EXPERIMENTAL
Melting points were determined on a Kofler block and are uncorrected. Optical rotation was
measured in chloroform (unless stated otherwise) on an automatic polarimeter ETL-NPL (Bendix–
Ericsson) with accuracy of ±2°. Infrared spectra were recorded in chloroform on a PE 684 (Perkin–
Elmer) spectrometer; wavenumbers are given in cm–1. Proton and 13C NMR spectra were taken on an
FT spectrometer Varian UNITY-500 (1H at 500 MHz, 13C at 125.7 MHz) in deuteriochloroform (un-
less stated otherwise) with tetramethylsilane as internal standard (for the 13C NMR data δ(CDCl3) =
77.00 ppm). In the 13C NMR spectra the number of directly bonded hydrogen atoms was determined
from proton decoupled “attached proton test” spectra. Values of chemical shifts (δ-scale) and coup-
1
ling constants (Hz) in the H NMR spectra were obtained by first order analysis and the assignment
was confirmed by H,H-COSY spectra. Proton NMR spectra of compounds 14b and 19b were
measured at 80 MHz on an FT spectrometer Tesla BS 587A. Fast atom bombardment mass spectra
(FAB-MS) were taken on a ZAB-EQ instrument (VG Analytical Ltd., Manchester), electron impact
mass spectra (EI-MS) on an INCOS 50 instrument (Finnigan MAT), ionizing electron energy 70 eV.
Methyl esters were prepared by treatment of the corresponding acids with ethereal solution of
diazomethane. Column chromatography was performed on silica gel Silpearl (Kavalier, Votice).
Purity of the samples was checked by thin-layer chromatography (TLC) on DC-Alufolien Kieselgel
60 foils (Merck). Spots were detected by spraying with 10% sulfuric acid and subsequent heating.
Preparative TLC was carried out on silica gel 60G (Merck); spots were detected with UV light (254 nm)
after spraying with 0.3% solution of morin in methanol. Analytical samples were dried over phos-
phorus pentoxide at 100 °C under diminished pressure.
28-Acetoxy-20(29)-lupen-3-one Oxime (14a)
A solution of ketone13 13a (9.7 g, 20 mmol) and hydroxylamine hydrochloride (7.0 g, 100 mmol) in
pyridine (50 ml) was heated at 80 °C for 1 h and then set aside at room temperature overnight. The
reaction mixture was poured into water, the formed precipitate was collected, washed with water,
dissolved in ether and the solution was dried over sodium sulfate. Purification by chromatography on
a column of silica gel in benzene–ether (10 : 1) and crystallization from chloroform–methanol af-
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

1788
Klinotova et al.:
forded oxime 14a (8.1 g, 81%), m.p. 203–204 °C, [α]D –7.5° (c 0.51). Reported16 m.p. 205–208 °C.
IR spectrum: 3 586, 3 264 broad, 1 723, 1 640, 1 388, 1 367, 1 246, 919, 892.
28-Acetoxylupan-3-one Oxime (14b)
Using the procedure described for oxime 14a, ketone13 13b (5.0 g, 10 mmol) was converted into
oxime 14b (4.1 g, 80%), m.p. 230–233 °C (chloroform–ethanol), [α]D –35° (c 0.52). Reported16 m.p.
219–220 °C. IR spectrum: 3 580, 3 270b, 1 724, 1 388, 1 367, 1 249, 914. 1H NMR spectrum (80 MHz):
0.76 d, 3 H, J = 6.5; 0.84 d, 3 H, J = 6.5; 0.94 s, 6 H; 1.05 s, 3 H; 1.07 s, 3 H and 1.14 s, 3 H (7 × CH3);
2.05 s, 3 H (CH3COO); 2.23 ddd, 1 H, J = 15.5; 11.5 and 5.5 (H-2β); 3.00 ddd, 1 H, J = 15.5, 5.5
and 3.5 (H-2α); 3.82 d, 1 H, J = 11 (H-28a); 4.26 d, 1 H, J = 11 (H-28b). EI-MS, m/z (%): 499 (M+, 19),
482 (67), 426 (10), 422 (12), 398 (20), 357 (20), 235 (68), 191 (37), 43 (100).
28-Hydroxy-3,4-secolupa-4(23),20(29)-diene-3-nitrile (18a)
Nitrile 19a (190 mg, 0.4 mmol) was hydrolyzed by treatment with 2.5% solution of potassium hy-
droxide in benzene–ethanol (1 : 1; 8 ml) at room temperature for three days. The reaction mixture
was dissolved in ether, washed with water and the ethereal solution was dried over sodium sulfate.
Evaporation of the solvent gave the nitrile 18a (160 mg, 92%), m.p. 115–117 °C (ether–light petroleum),
[α]D +37° (c 0.47). IR spectrum: 3 624, 3 071, 2 243, 1 638, 893. EI-MS, m/z (%): 437 (M+, 32),
419 (15), 406 (100), 283 (32), 189 (39). For C30H47NO (437.7) calculated: 82.32% C, 10.82% H, 3.20% N;
found: 82.06% C, 10.64% H, 3.52% N.
28-Hydroxy-3,4-secolup-4(23)-ene-3-nitrile (18b)
Using the procedure described for nitrile 18a, nitrile 19b (200 mg, 0.4 mmol) was converted into
nitrile 18b (160 mg, 88%), m.p. 157–159 °C (ether), [α]D –5° (c 0.50). IR spectrum: 3 625, 3 071, 2 245,
1 633, 898. EI-MS, m/z (%): 439 (M+, 9), 424 (4.5), 421 (5), 408 (100), 191 (43). For C30H49NO
(439.7) calculated: 81.94% C, 11.23% H, 3.19% N; found: 81.71% C, 10.99% H, 3.48% N.
28-Acetoxy-3,4-secolupa-4(23),20(29)-diene-3-nitrile (19a)
A solution of oxime 14a (8.0 g, 16.2 mmol) and 4-toluenesulfonyl chloride (18 g, 94 mmol) in py-
ridine (150 ml) was refluxed for 3 h. After cooling, the reaction mixture was poured into water, the
precipitate was collected, washed with water and dissolved in chloroform. The solution was dried by
filtration through a layer of silica gel and the product was crystallized from chloroform–methanol to
give nitrile 19a (6.6 g, 85%), m.p. 121–122 °C, [α]D +24° (c 0.49). Reported16 m.p. 120–122 °C. IR
spectrum: 3 070, 2 247, 1 722, 1 632, 1 242, 895. EI-MS, m/z (%): 479 (M+, 34), 419 (21), 406 (27),
376 (18), 338 (16), 203 (62), 189 (33), 43 (100).
28-Acetoxy-3,4-secolup-4(23)-ene-3-nitrile (19b)
Using the procedure described for nitrile 19a, oxime 14b (4.7 g, 9.5 mmol) was converted into amor-
phous nitrile 19b (3.9 g, 86%), [α]D –8° (c 0.54). IR spectrum: 3 071, 2 244, 1 724, 1 633, 1 247,
1
900. H NMR spectrum (80 MHz): 0.78 d, 3 H, J = 6.5; 0.85 d, 3 H, J = 6.5; 0.85 s, 3 H; 0.97 s, 3 H,
and 1.09 s, 3 H (5 × CH3); 1.72 bs, 3 H (3 × H-24); 2.05 s, 3 H (CH3COO); 2.14–2.46 m, 2 H (2 × H-2);
3.83 d, 1 H, J = 11 (H-28a); 4.26 dd, 1 H, J = 11 and 1 (H-28b); 4.65 bd, 1 H, J = 2 (H-23a); 4.88 m,
1 H (H-23b). EI-MS, m/z (%): 481 (M+, 15), 421 (14), 408 (23), 378 (6), 340 (13), 289 (10),
229 (15), 203 (20), 191 (57), 43 (100).
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

Triterpenes
28-Hydroxy-3,4-secolupa-4(23),20(29)-dien-3-oic Acid (22a)
1789
A solution of nitrile 19a (6.0 g, 12.5 mmol) and sodium hydroxide (15 g, 0.4 mol) in ethylene glycol
(150 ml) was refluxed for 6 h. After cooling, the reaction mixture was poured into an excess of dilute
hydrochloric acid (1 : 4), the precipitate was collected, washed with water till neutral reaction of the
washings, dissolved in ether and the solution was dried over sodium sulfate. Repeated crystallization
from chloroform–heptane afforded acid 22a (4.7 g, 82%), m.p. 136–162 °C (decomp.), [α]D +36° (c 0.47).
IR spectrum: 3 514, 3 300–2 600, 3 071, 1 707, 1 640, 894. EI-MS, m/z (%): 456 (M+,17), 425 (25),
383 (26), 375 (23), 357 (60), 345 (41), 81 (100).
Methyl ester 23a: m.p. 171–173 °C (ether–light petroleum), [α]D +33° (c 0.38). IR spectrum: 3 621,
3 071, 1 728, 1 638, 1 438, 895. EI-MS, m/z (%): 470 (M+, 16), 439 (15), 389 (12), 383 (55),
371 (42), 359 (52), 189 (42), 81 (100). For C31H50O3 (470.7) calculated: 79.10% C, 10.71% H;
found: 79.18% C, 10.98% H.
28-Hydroxy-3,4-secolup-4(23)-en-3-oic Acid (22b)
Using the procedure described for acid 22a, nitrile 19b (3.1 g, 6.5 mmol) was converted into acid 22b
(2.7 g, 91%), m.p. 137–165 °C (decomp.) (ether–light petroleum), [α]D –3° (c 0.49). IR spectrum: 3 621,
3 300–2 700, 1 705, 1 638, 895. EI-MS, m/z (%): 458 (M+,23), 427 (37), 385 (34), 359 (100),
347 (22), 303 (13).
Methyl ester 23b: m.p. 166–168 °C (ether), [α]D –5° (c 0.59). Reported17 m.p. 122 °C (ethyl acetate).
IR spectrum: 3 621, 1 729, 1 638, 1 443, 895. EI-MS, m/z (%): 472 (29), 441 (24), 385 (88),
373 (85), 361 (68), 191 (65), 189 (42), 109 (100). For C31H52O3 (472.8) calculated: 78.76% C,
11.09% H; found: 79.02% C, 11.23% H.
28-Acetoxy-3,4-secolupa-4(23),20(29)-dien-3-oic Acid (24a)
Hydroxy acid 22a (350 mg, 77 mmol) was acetylated with acetic anhydride–pyridine mixture (3 : 2,
5 ml) at room temperature overnight. The reaction mixture was decomposed with excess of water, the
product was taken up in ether, the ethereal solution was washed with dilute hydrochloric acid, 5%
solution of sodium hydrogen carbonate and with water and dried over sodium sulfate. Crystallization
from chloroform–heptane afforded acid 24a (310 mg, 81%), m.p. 227–230 °C, [α]D +26° (c 0.45). IR
spectrum: 3 514, 3 071, 3 300–2 600, 1 729, 1 709, 1 640, 894. EI-MS, m/z (%): 498 (M+, 8),
438 (5), 425 (22), 357 (62), 187 (25), 189 (24), 43 (100). For C32H50O4 (498.8) calculated: 77.06% C,
10.10% H; found: 77.22% C, 10.21% H.
Methyl ester 25a: amorphous, [α]D +23° (c 0.56). IR spectrum: 3 071, 1 728, 1 640, 1 437, 1 246,
892. EI-MS, m/z (%): 512 (M+, 16), 431 (5), 425 (31), 371 (86), 201 (30), 187 (38), 43 (100).
28-Acetoxy-3,4-secolup-4(23)-en-3-oic Acid (24b)
Using the procedure described for acid 24a, acid 22b (100 mg, 22 mmol) was converted into acid
24b (100 mg, 92%), m.p. 199–201 °C (ether), [α]D –6° (c 0.69). IR spectrum: 3 509, 3 069, 3 300–2 600,
1 730, 1 708, 1 636, 1 246, 897. EI-MS, m/z (%): 500 (M+, 23), 440 (10), 427 (52), 367 (23),
359 (28), 191 (15), 43 (100). For C32H52O4 (500.8) calculated: 76.75% C, 10.47% H; found: 76.54% C,
10.52% H.
Methyl ester 25b: amorphous, [α]D –7° (c 0.57). IR spectrum: 1 730, 1 634, 1 437, 1 245, 897.
EI-MS, m/z (%): 514 (M+, 4), 454 (1.5), 441 (2.5), 427 (9), 373 (38), 191 (21), 43 (100).
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

1790
Klinotova et al.:
3,4-Secolupa-4(23),20(29)-diene-3,28-diol (32a)
A mixture of methyl ester 23a (650 mg, 1.4 mmol) and lithium aluminium hydride (500 mg, 13.2 mmol)
in ether (200 ml) was refluxed for 1 h. The reaction mixture was then decomposed with ethyl acetate,
water and dilute sulfuric acid. The ethereal layer was washed with brine and dried over sodium sul-
fate. The solvent was evaporated and the residue was crystallized from ether–light petroleum to give
diol 32a (550 mg, 90%), m.p. 212–213 °C, [α]D +47° (c 0.49). Reported18 m.p. 212–213 °C, [α]D
+45° (methanol). IR spectrum: 3 623, 1 637, 893. EI-MS, m/z (%): 442 (M+, 27), 411 (11), 383 (24),
361 (25), 343 (50), 331 (41), 201 (50), 85 (100).
Diacetate 33a was prepared by acetylation of diol 32a (100 mg, 0.22 mmol) according to the pro-
cedure described for the 28-acetoxy acid 24a. Chromatographic purification of the crude product af-
forded amorphous diacetate 33a (100 mg, 83%), [α]D +30° (c 0.37). IR spectrum: 1 724, 1 249.
EI-MS, m/z (%): 526 (M+, 5), 466 (2), 453 (2), 445 (2), 425 (7), 385 (35), 325 (12), 43 (100). Ref.19
reports only the 13C NMR spectrum.
Treatment of diol 32a (530 mg, 1.2 mmol) with acetic anhydride (0.6 ml, 6.3 mmol) in pyridine
(15 ml) at –30 °C for 4 h, followed by the above-described workup, afforded a mixture of products
which on chromatographic separation afforded: diacetate 33a (80 mg, 13%), the starting diol 32a
(350 mg, 66%) and a chromatographically unseparable mixture of monoacetates 34a and 35a (1:1
according to 1H NMR spectrum; 90 mg, 18%). EI-MS of the mixture, m/z (%): 484 (M+, 25), 453
(14, corresponds to isomer 34a), 425 (11, 35a), 424 (5), 403 (12), 411 (5, 35a), 385 (49), 383 (100, 34a),
343 (66).
General Procedure for Preparation of Tetra-O-acetyl-β-D-glucopyranosyl Derivatives
A. In acetonitrile in the presence of mercury(II) cyanide: A mixture of the aglycone, 2,3,4,6-tetra-
O-acetyl-α-D-glucopyranosyl bromide (further only acetobromoglucose) and mercury(II) cyanide in
anhydrous acetonitrile (or its mixture with ether or chloroform) was refluxed. The amount of the
starting aglycone was followed by TLC and, as required, further acetobromoglucose and mercury(II)
cyanide was added. After completion of the reaction, the reaction mixture was diluted with ether, the
ethereal solution was washed with water and dried over sodium sulfate. The solvent was evaporated
and the residue subjected to preparative TLC in light petroleum–ether (1 : 1 to 2 : 3, according to
polarity of the compounds separated).
B. In dichloromethane in the presence of silver silicate: A solution of acetobromoglucose in di-
chloromethane was added under argon to a mixture of the aglycone, silver silicate and calcined mole-
cular sieve (4 Å) in dichloromethane. The reaction mixture was stirred in the dark at room
temperature for 24 h and then without work-up subjected to column chromatography on silica gel in
toluene–ether (4 : 1).
Methyl 3β-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyloxy)-lup-20(29)-en-28-oate (4a)
A mixture of hydroxy derivative 2a (200 mg, 0.4 mmol), acetobromoglucose (650 mg, 1.6 mmol),
mercury(II) cyanide (215 mg, 0.9 mmol) and acetonitrile (10 ml) was refluxed for 34 h to give glu-
coside acetate 4a (180 mg, 53%), m.p. 120–123 °C (ether–light petroleum), [α]D 0° (c 0.56). IR spec-
trum: 1 754, 1 740, 1 720, 1 640, 1 433, 1 222, 909. EI-MS, m/z (%): 800 (M+, 0.3), 741 (0.3),
740 (0.3), 537 (9), 469 (4), 453 (5), 393 (13), 331 (20), 189 (61), 169 (98), 109 (96), 43 (100). For
C45H68O12 (801.0) calculated: 67.48% C, 8.56% H; found: 67.28% C, 8.69% H. Substantial amount
of the starting hydroxy derivative 2a (50 mg, 25%) was recovered.
Collect. Czech. Chem. Commun. (Vol. 62) (1997)

Triterpenes
Methyl 3β-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyloxy)-lupan-28-oate (4b)
1791
A mixture of hydroxy derivative 2b (200 mg, 0.4 mmol), acetobromoglucose (650 mg, 1.6 mmol),
mercury(II) cyanide (215 mg, 0.9 mmol), acetonitrile (10 ml) and anhydrous chloroform (8 ml) was
refluxed for 36 h. The reaction gave glucoside acetate 4b (95 mg, 28%), m.p. 122–126 °C (chloro-
form–methanol), [α]D –16° (c 0.71). IR spectrum: 1 753, 1 740, 1 720, 1 433, 1 225. EI-MS, m/z(%):
802 (M+, 0.01), 743 (1), 537 (4), 472 (4), 470 (5), 455 (78), 411 (12), 395 (25), 331 (15), 275 (12),
271 (4), 189 (52), 169 (72), 109 (90), 43 (100). For C45H70O12 (803.1) calculated: 67.31% C, 8.79% H;
found: 67.15% C, 8.52% H. Great amount of the starting compound 2b (120 mg, 60%) was re-
covered.
2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl 3β-Acetoxy-lup-20(29)-en-28-oate (7a)
A mixture of acid 3a (200 mg, 0.4 mmol), acetobromoglucose (420 mg, 1 mmol), mercury(II)
cyanide (250 mg, 1 mmol), acetonitrile (10 ml) and anhydrous ether (7 ml) was refluxed for 12.5 h.
The reaction afforded amorphous derivative 7a (200 mg, 60%). [α]D +18° (c 0.55). IR spectrum: 1 754,
1 744, 1 720, 1 641, 902. EI-MS, m/z (%): 828 (M+, 0.1), 768 (0.2), 725 (0.1), 331 (66), 271 (15),
189 (12), 169 (100), 109 (37), 43 (95).
2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl 3β-Acetoxylupan-28-oate (7b)
A mixture of acid 3b (200 mg, 0.4 mmol), acetobromoglucose (550 mg, 1.3 mmol), mercury(II)
cyanide (150 mg, 0.6 mmol), acetonitrile (10 ml) and anhydrous ether (10 ml) was refluxed for 16 h.
The reaction afforded amorphous derivative 7b (78 mg, 24%). [α]D +1° (c 0.67). IR spectrum: 1 755,
1 745, 1 722, 1 252. EI-MS, m/z(%): 830 (M+, 0.1), 770 (1), 755 (0.4), 727 (0.3), 455 (3), 439 (3),
411 (1), 395 (4), 331 (72), 271 (19), 189 (19), 169 (100), 109 (42), 43 (73). Substantial amount of
the starting acid 3b (80 mg, 40%) was recovered.
2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl 3β-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyloxy)-
lup-20(29)-en-28-oate (10a)
A mixture of acid 1a (200 mg, 0.4 mmol), acetobromoglucose (650 mg, 1.6 mmol), mercury(II)
cyanide (405 mg, 1.6 mmol), acetonitrile (20 ml), anhydrous ether (8 ml) and anhydrous chloroform (5 ml)
was refluxed for 19 h to afford derivative 10a (40 mg, 8%), m.p. 225–231 °C (dichloromethane–ether),
[α]D +1° (c 0.46). IR spectrum: 1 755 broad, 1 642, 1 218, 908. EI-MS, m/z (%): 785 (M+ –331, 0.1),
769 (0.1), 537 (0.1), 331 (28), 271 (10), 169 (100), 109 (41), 43 (49). For C58H84O21 (1117.3) calcu-
lated: 62.35% C, 7.58% H; found: 62.15% C, 7.22% H. Great amount of the starting acid 1a (150 mg,
75%) was recovered.
28-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyloxy)lup-20(29)-en-3-one (16a)
Reaction of hydroxy derivative 12a (220 mg, 0.5 mmol), acetobromoglucose (600 mg, 1.5 mmol),
molecular sieve (800 mg) and silver silicate (500 mg) in dichloromethane (20 ml) afforded amor-
phous orthoacetate 15a (120 mg, 31%) which on attempted purification decomposed to give hydroxy
derivative 12a and two tetra-O-acetyl derivatives of glucose (according to TLC). Also isolated was
glucoside acetate 16a (50 mg, 13%), m.p. 130–131 °C (ether–heptane), [α]D +7° (c 0.34). IR spec-
trum: 1 755, 1 740, 1 698, 1 638, 1 218, 908. EI-MS, m/z (%): 770 (M+, 0.3), 451 (0.3), 409 (10),
331 (44), 271 (10), 169 (100), 109 (40), 43 (53). For C44H66O11 (771.0) calculated: 68.55% C,
8.64% H; found: 68.60% C, 8.48% H.
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28-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyloxy)lupan-3-one (16b)
A mixture of hydroxy derivative 12b (50 mg, 0.1 mmol), acetobromoglucose (110 mg, 0.3 mmol),
mercury(II) cyanide (30 mg, 0.12 mmol) and acetonitrile (5 ml) was refluxed for 12 h. The reaction
afforded glucoside acetate 16b (45 mg, 52%), m.p. 173.5–175 °C (ether–light petroleum), [α]D –6° (c 0.56).
IR spectrum: 1 754, 1 740, 1 698, 1 224. EI-MS, m/z (%): 772 (M, 0.2), 540 (0.4), 480 (0.4),
453 (0.6), 440 (0.6), 411 (18), 331 (52), 169 (100), 109 (49), 43 (85). For C44H68O11 (773.0) calcu-
lated: 68.37% C, 8.87% H; found: 68.58% C, 9.03% H.
28-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyloxy)-3,4-seco-lupa-4(23),20(29)-diene-3-nitrile (20a)
A mixture of hydroxy derivative 18a (150 ml, 0.3 mmol), acetobromoglucose (420 mg, 1 mmol),
mercury(II) cyanide (260 mg, 1 mmol) and acetonitrile (10 ml) was refluxed for 10 h. The reaction
afforded glucoside acetate 20a (172 mg, 65%), m.p. 182–184 °C (methanol), [α]D +2° (c 0.57). IR
spectrum: 3 068, 2 247, 1 754 broad, 1 639, 1 219, 895. EI-MS, m/z (%): 767 (M+, 5), 465 (0.6),
448 (0.6), 419 (11), 406 (21), 331 (49), 271 (14), 169 (100), 109 (42), 43 (53). For C44H65NO10
(768.0) calculated: 68.81% C, 8.53% H; found: 68.52% C, 8.40% H.
28-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyloxy)-3,4-secolup-4(23)-ene-3-nitrile (20b)
A mixture of hydroxy derivative 18b (130 mg, 0.3 mmol), acetobromoglucose (375 mg, 0.9 mmol),
mercury(II) cyanide (225 mg, 0.9 mmol) and acetonitrile (10 ml) was refluxed for 10 h to give glu-
coside acetate 20b (140 mg, 62%), m.p. 198–202 °C (chloroform–methanol), [α]D –2° (c 0.52). IR
spectrum: 2 247, 1 754b, 1 634, 1 218, 899. EI-MS, m/z (%): 769 (M+, 2), 577 (0.3), 450 (0.1),
422 (10), 408 (23), 331 (66), 271 (12), 169 (100), 109 (48), 43 (56). For C44H67NO10 (770.0) calcu-
lated: 68.63% C, 8.77% H; found: 68.20% C, 8.59% H.
Methyl 28-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyloxy)-3,4-secolupa-4(23),20(29)-dien-3-oate (27a)
A) A mixture of hydroxy derivative 23a (200 mg, 0.4 mmol), acetobromoglucose (265 mg, 0.6 mmol),
mercury(II) cyanide (165 mg, 0.7 mmol) and acetonitrile (10 ml) was refluxed for 7 h. The reaction
afforded amorphous glucoside acetate 27a (220 mg, 65%), [α]D +5° (c 0.49). IR spectrum: 1 754,
1 740, 1 635, 1 436, 1 227, 898. EI-MS, m/z (%): 800 (M+, 0.1), 452 (3), 439 (4), 371 (6),
169 (100), 109 (37), 43 (51). The reaction further afforded compound 25a (30 mg, 14%), identical
with an authentic sample.
B) Reaction of a mixture of hydroxy derivative 23a (400 mg, 0.8 mmol), molecular sieve (1 g),
silver silicate (650 mg) and acetobromoglucose (700 mg, 1.7 mmol) in dichloromethane (15 ml) af-
forded orthoacetate 26a (200 mg, 29%) and amorphous glucoside acetate 27a (90 mg, 13%), identical
with the compound prepared by procedure A.
Orthoacetate 26a: m.p. 100–102 °C (heptane), [α]D +29° (c 0.49). IR spectrum: 1 746, 1 640, 1 435,
1 221, 893. For 1H NMR spectrum see Table II. After standing for 2 days in chloroform at room
temperature, the orthoacetate 26a decomposed to give a mixture which, according to the 1H a 13C NMR
spectra displayed signals of the hydroxy derivative 23a (see Tables II and VI), 1,3,4,6-tetra-O-acetyl-
α-D-glucopyranose and 2,3,4,6-tetra-O-acetyl-α-D-glucopyranose in the ratio 3 : 2 : 1.
1,3,4,6-Tetra-O-acetyl-α-D-glucopyranose: 1H NMR spectrum (500 MHz): 2.053 s, 3 H; 2.087 s,
3 H; 2.109 s, 3 H and 2.208 s, 3 H (4 × CH3); 3.66 d, 1 H, J = 9 (OH); 3.90 ddd, 1 H, J = 10, 9
and 4 (H-2); 4.03 ddd, 1 H, J = 10, 4 and 2 (H-5); 4.06 dd, 1 H, J = 12.5 and 2 (H-6a); 4.29 dd, 1 H,
J = 12.5 and 4 (H-6b); 5.11 t, 1 H, J = 10 and 10 (H-4); 5.27 t, 1 H, J = 10 and 10 (H-3); 6.24 d,
1 H, J = 4 (H-1). 13C NMR spectrum: 20.5–20.9 (4 × CH3), 61.57 (C-6), 67.35 (C-4), 69.66 (C-5),
69.89 (C-2), 73.19 (C-3), 91.32 (C-1), 169–172 (4 × C=O).
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2,3,4,6-Tetra-O-acetyl-α-D-glucopyranose: 1H NMR spectrum (500 MHz): 2.025 s, 3 H; 2.042 s,
3 H; 2.093 s, 3 H and 2.104 s, 3 H (4 × CH3); 3.05 dd, 1 H, J = 4 and 1 (OH); 4.13 m, 1H (H-6a);
4.23–4.25 m, 2 H (H-5 and H-6b); 4.92 ddd, 1 H, J = 10, 4 and 1 (H-2); 5.10 t, 1 H, J = 10 and 10
(H-4); 5.54 t, 1 H, J = 10 and 10 (H-3); 5.48 t, 1 H, J = 4 and 4 (H-1). 13C NMR spectrum: 20.5–
20.9 (4 × CH3), 61.90 (C-6), 67.29 (C-5), 68.44 (C-4), 69.81 (C-3), 71.01 (C-2), 90.21 (C-1), 169-
172 (4 × C=O).
Methyl 28-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyloxy)-3,4-secolup-4(23)-en-3-oate (27b)
A mixture of hydroxy derivative 23b (200 mg, 0.4 mmol), acetobromoglucose (355 mg, 0.9 mmol),
mercury(II) cyanide (225 mg, 0.9 mmol) and acetonitrile (10 ml) was refluxed for 7 h. The reaction
afforded glucoside acetate 27b (230 mg, 68%), m.p. 165–166 °C (methanol), [α]D –10° (c 0.57). IR
spectrum: 1 507b, 1 635, 1 436, 1 227, 898. EI-MS, m/z (%): 802 (M+, 0.5), 770 (0.2), 715 (0.5),
577 (0.3), 471 (1), 441 (2), 373 (20), 331 (100), 271 (19), 169 (89), 109 (42), 43 (55). For
C45H70O12 (803.1) calculated: 67.31% C, 8.79% H; found: 66.95% C, 8.53% H.
2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl 28-Acetoxy-3,4-secolupa-4(23),20(29)-dien-3-oate (30a)
A mixture of acid 24a (220 mg, 0.4 mmol), acetobromoglucose (545 mg, 1.3 mmol), mercury(II)
cyanide (225 mg, 0.9 mmol) and acetonitrile (10 ml) was refluxed for 26 h. The reaction afforded
amorphous derivative 30a (180 mg, 49%), [α]D +12° (c 0.55). IR spectrum: 1 758, 1 740, 1 722, 1 639,
1 226, 895. EI-MS, m/z (%): 828 (M+, 0.01), 768 (0.01), 755 (0.01), 497 (0.3), 481 (0.9), 437 (0.5),
425 (0.5), 421 (1.3), 357 (0.9), 331 (45), 271 (12), 169 (100), 109 (61), 43 (52). Part of the starting
acid 24a (90 mg, 41%) was recovered.
3,28-Bis(2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyloxy)-3,4-secolupa-4(23),20(29)-diene (36a) and
3-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyloxy)-3,4-secolupa-4(23),20(29)-dien-28-ol (38a)
A mixture of diol 32a (200 mg, 0.5 mmol), acetobromoglucose (560 mg, 1.4 mmol), mercury(II)
cyanide (350 mg, 1.4 mmol) and acetonitrile (20 ml) was refluxed for 7 h. The reaction afforded
amorphous glucoside acetate 38a (70 mg, 20%). [α]D +8° (c 0.94). IR spectrum: 3 624, 1 753, 1 742,
1 638, 1 226, 892. EI-MS, m/z (%): 772 (M+, 0.5), 741 (0.5), 691 (0.4), 673 (2), 661 (2), 643 (0.4),
511 (0.4), 383 (10), 331 (71), 271 (18), 170 (76), 169 (43), 109 (81), 43 (100).
Further isolated product was derivative 36a (300 mg, 60%), m.p. 225–227 °C (methanol), [α]D –6°
(c 0.48). IR spectrum: 1 754, 1 740, 1 639, 1 219, 892. EI-MS, m/z (%): 772 (M+ – 330, 0.1),
754 (0.2), 741 (0.6), 713 (0.2), 673 (0.7), 643 (0.2), 511 (0.1), 423 (0.6), 383 (0.5), 365 (0.7),
331 (61), 271 (18), 169 (100), 109 (39), 43 (37). For C58H86O20 (1103.3) calculated: 63.14% C,
7.86% H; found: 62.88% C, 7.64% H.
Also isolated was a mixture of isomeric glucoside acetates 40a and 41a (45 mg, 12%). EI-MS of
the mixture, m/z (%): 814 (M+, 1), 754 (0.2), 741 (0.2, isomer 40a), 733 (0.2), 713 (0.2, 41a), 673 (1),
453 (4, 41a), 425 (4, 40a), 331 (82), 169 (100), 109 (39), 43 (68).
Dimethyl 28-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyloxy)-2,3-secolup-20(29)-ene-2,3-dioate (46a)
A) A mixture of hydroxy derivative 43a (200 mg, 0.4 mmol), acetobromoglucose (410 mg, 1 mmol),
mercury(II) cyanide (200 mg, 0.8 mmol) and acetonitrile (10 ml) was refluxed for 16 h. The reaction
afforded glucoside acetate 46a (140 mg, 43%), m.p. 169–171 °C (methanol), [α]D –1° (c 0.46). IR
spectrum: 1 753, 1 740, 1 720, 1 639, 1 433, 1 227, 888. EI-MS, m/z (%): 846 (M+, 0.2), 815 (0.1),
787 (0.25), 772 (0.3), 745 (0.2), 671 (0.5), 577 (0.4), 515 (0.3), 331 (70), 271 (10), 169 (100),
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109 (32), 43 (53). For C46H70O14 (847.1) calculated: 65.23% C, 8.33% H; found: 65.02% C, 8.13%
H. As a side product was isolated acetate 44a (40 mg, 18%), identical with an authentic13 sample.
Also a part of the starting hydroxy derivative 43a (60 mg, 30%) was recovered.
B) Reaction of hydroxy derivative 43a (300 mg, 0.6 mmol) with acetobromoglucose (600 mg, 1.5 mmol)
in the presence of silver silicate (700 mg) and a molecular sieve (1 g) v dichloromethane (20 ml)
afforded orthoacetate 45a (160 mg, 33%), m.p. 145–148 °C (ether), [α]D +13° (c 1.05). IR spectrum:
1 720, 1 639, 1 433, 1 224, 888. For the 1H NMR spectrum see Table III. In chloroform solution,
ortho acetate 45a decomposed to give a mixture of hydroxy derivative 43a and two tetra-O-acetyl
derivatives of glucose (according to TLC). Further isolated product was the glucoside acetate 46a
(105 mg, 21 %), identical with the sample prepared according to procedure A.
Dimethyl 28-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyloxy)-2,3-secolupane-2,3-dioate (46b)
A) A mixture of hydroxy derivative 43b (210 mg, 0.4 mmol), acetobromoglucose (420 mg, 1 mmol),
mercury(II) cyanide (200 mg, 0.8 mmol) and acetonitrile (10 ml) was refluxed for 16 h. The reaction
afforded glucoside acetate 46b (150 mg, 44%), m.p. 190–194 °C (chloroform–methanol), [α]D –18°
(c 0.53). IR spectrum: 1 753, 1 740, 1 720, 1 434, 1 227, 908. EI-MS, m/z (%): 848 (M+, 0.1),
818 (0.5), 774 (0.2), 715 (0.3), 673 (0.2), 577 (0.3), 517 (0.3), 331 (87), 271 (13), 169 (100),
109 (30), 43 (42). For C46H72O14 (849.1) calculated: 65.07% C, 8.55% H; found: 64.85% C, 8.60% H.
Further isolated product was the acetate 44b (7 mg, 3%), identical with an authentic sample13
Also a part of the starting hydroxy derivative 43b (85 mg, 43%) was recovered.
.
B) Reaction of hydroxy derivative 43b (300 mg, 0.6 mmol) with acetobromoglucose (600 mg, 1.5 mmol)
in the presence of silver silicate (700 mg) and a molecular sieve (1 g) in dichloromethane (20 ml)
afforded orthoacetate 45b (290 mg, 59%), m.p. 83–90/150–153 °C (ether), [α]D –15° (c 0.52). IR
spectrum: 1 720, 1 435, 1 222. In chloroform solution, orthoacetate 45b decomposed to give a mix-
ture of hydroxy derivative 43b and two tetra-O-acetylderivatives of glucose (TLC).
General Procedure for Deacetylation of Tetra-O-acetyl-β-D-glucopyranosyl Derivatives
The glucoside acetate was dissolved in anhydrous methanol (2–6 ml, according to the amount and
solubility of the compound), 5% methanolic solution of sodium methoxide (0.1–0.2 ml) was added
and the solution was set aside at room temperature. The reaction time ranged from 1 to 3 h and the
reaction was monitored by TLC. If the glucoside acetate was insoluble in methanol, the reaction mix-
ture was stirred until the acetate dissolved. The work-up consisted in neutralization of the reaction
mixture with 10% acetic acid and evaporation at 30 °C under diminished pressure. The residue was
purified by preparative TLC in chloroform–methanol (9 : 1).
Methyl 3β-(β-D-Glucopyranosyloxy)-lup-20(29)-en-28-oate (5a)
Using the general procedure, glucoside acetate 4a (120 mg, 0.15 mmol) was converted into glucoside
5a (70 mg, 74%), m.p. 197–200/248–250 °C (methanol), [α]D –3° (c 0.66, methanol). FAB-MS, m/z:
655 (M + Na), 633 (M + H), 471, 453, 393.
Methyl 3β-(β-D-Glucopyranosyloxy)lupan-28-oate (5b)
Glucoside acetate 4b (21 mg, 0.03 mmol) was converted into glucoside 5b (14 mg, 84%), m.p. 265–269 °C
(chloroform–methanol), [α]D –21° (c 0.38, methanol). FAB-MS, m/z: 657 (M + Na), 635 (M + H),
473, 455, 395.
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Triterpenes
β-D-Glucopyranosyl 3β-Acetoxylup-20(29)-en-28-oate (8a)
1795
Glucoside acetate 7a (170 mg, 0.21 mmol) was converted into amorphous glucoside 8a (110 mg,
81%), [α]D +4° (c 0.59, methanol). FAB-MS, m/z: 683 (M + Na), 499, 453, 439, 393.
β-D-Glucopyranosyl 3β-Acetoxylupan-28-oate (8b)
Glucoside acetate 7b (85 mg, 0.09 mmol) was converted into amorphous glucoside 8b (58 mg, 86%),
[α]D –13° (c 0.53, methanol). FAB-MS, m/z: 685 (M + Na), 501, 455, 441, 395.
β-D-Glucopyranosyl 3β-Hydroxylup-20(29)-en-28-oate (9a)
Acetate 8a (20 mg, 0.03 mmol) was dissolved in a 1.5% solution of sodium hydroxide (0.75 mmol)
in methanol (2 ml) and the solution was warmed to 50 °C for 30 min. After cooling, the reaction
mixture was applied onto a preparative TLC plate and chromatographed in chloroform–methanol (9 : 1).
Yield 15 mg (80%) of amorphous glucoside 9a, [α]D 0° (c 0.16, methanol); reported26 m.p. 213–216 °C,
[α]D –2° (ethanol). FAB-MS, m/z: 641 (M + Na), 619 (M + H), 601, 457, 439, 411, 393.
β-D-Glucopyranosyl 3β-Hydroxylupan-28-oate (9b)
Using the same procedure and the same chromatography conditions as described for the preparation
of glucoside 9a, conversion of acetate 8b (20 mg, 0.03 mmol) afforded glucoside 9b (17 mg, 91%),
m.p. 243–244 °C (methanol), [α]D –21° (c 0.17, methanol). FAB-MS, m/z: 643 (M + Na), 621 (M + H),
603, 459, 441, 413, 395.
β-D-Glucopyranosyl 3β-(β-D-Glucopyranosyloxy)lup-20(29)-en-28-oate (11a)
Deacetylation of glucoside acetate 10a (20 mg, 0.02 mmol) afforded glucoside 11a (9 mg, 64%),
m.p. 211–212 °C (methanol), [α]D –5° (c 0.42, methanol). Reported25 m.p. 209–211 °C, [α]D –5°
(ethanol). FAB-MS, m/z: 803 (M + Na), 619, 601, 439.
28-(β-D-Glucopyranosyloxy)lup-20(29)-en-3-one (17a)
Deacetylation of glucoside acetate 16a (30 mg, 0.04 mmol) afforded glucoside 17a (16 mg, 53%),
m.p. 202–206 °C (methanol), [α]D +2° (c 0.29, methanol). FAB-MS, m/z: 625 (M + Na), 603 (M + H),
441, 423.
28-(β-D-Glucopyranosyloxy)lupan-3-one (17b)
Deacetylation of glucoside acetate 16b (30 mg, 0.04 mmol) afforded glucoside 17b (20 mg, 85%),
m.p. 210–211 °C (methanol), [α]D –15° (c 0.51, methanol). FAB-MS, m/z: 627 (M + Na), 605 (M + H),
443, 425.
28-(β-D-Glucopyranosyloxy)-3,4-secolupa-4(23),20(29)-diene-3-nitrile (21a)
Deacetylation of glucoside acetate 20a (165 mg, 0.20 mmol) afforded glucoside 21a (117 mg, 91%),
m.p. 147–149/218–225 °C (methanol), [α]D +4° (c 0.55, methanol). FAB-MS, m/z: 600 (M + H),
438, 420.
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28-(β-D-Glucopyranosyloxy)-3,4-secolup-4(23)-ene-3-nitrile (21b)
Deacetylation of glucoside acetate 20b (12 mg, 0.01 mmol) afforded glucoside 21b (8 mg, 85%),
m.p. 130–133 °C (methanol), [α]D –16° (c 0.35, methanol). FAB-MS, m/z: 602 (M + H), 440, 422.
Methyl 28-(β-D-Glucopyranosyloxy)-3,4-secolupa-4(23),20(29)-dien-3-oate (28a)
Deacetylation of glucoside acetate 27a (158 mg, 0.20 mmol) afforded glucoside 28a (86 mg, 69%),
m.p. 149–151 °C (methanol), [α]D 0° (c 0.50, methanol). FAB-MS, m/z: 655 (M + Na), 633 (M + H),
471, 453.
Methyl 28-(β-D-Glucopyranosyloxy)-3,4-secolup-4(23)-en-3-oate (28b)
Deacetylation of glucoside acetate 27b (170 mg, 0.21 mmol) afforded glucoside 28b (125 mg, 93%),
m.p. 135–138 °C (methanol), [α]D –23° (c 0.64, methanol). FAB-MS, m/z: 657 (M + Na), 473, 455.
28-(β-D-Glucopyranosyloxy)-3,4-secolupa-4(23),20(29)-dien-3-oic Acid (29a)
Glucoside 28a (96 mg, 0.15 mmol) was dissolved in a 3% solution of sodium hydroxide (6 mmol) in
methanol (8 ml) and the solution was set aside at room temperature for 2 days. The reaction mixture
was acidified with 20% acetic acid and the solvents were evaporated at 30 °C under diminished
pressure. The residue was extracted with a chloroform–methanol mixture (5 : 1) and the extract was
filtered through a layer of silica gel. Yield 86 mg (91%) of glucoside 29a, m.p. 287–290 °C (chloro-
form–ethanol), [α]D +3° (c 0.55, methanol). FAB-MS, m/z: 641 (M + Na), 619 (M + H), 457, 439.
28-(β-D-Glucopyranosyloxy)-3,4-secolup-4(23)-en-3-oic Acid (29b)
Using the same procedure as described for the preparation of glucoside 29a, glucoside 28b (70 mg,
0.11 mmol) was converted into amorphous glucoside 29b (60 mg, 88%), [α]D –16° (c 0.51, methanol).
FAB-MS, m/z: 643 (M + Na), 459, 441.
3,28-Bis(β-D-glucopyranosyloxy)-3,4-secolupa-4(23),20(29)-diene (37a)
Deacetylation of glucoside acetate 36a (180 mg, 0.16 mmol) afforded amorphous glucoside 37a (95 mg,
76%), [α]D 0° (c 0.48, methanol).
3-(β-D-Glucopyranosyloxy)-3,4-secolupa-4(23),20(29)-dien-28-ol (39a)
Deacetylation of glucoside acetate 38a (27 mg, 0.03 mmol) afforded amorphous glucoside 39a (14 mg,
66%), [α]D +19° (c 0.48, methanol). FAB-MS, m/z: 627 (M + Na), 605 (M + H), 437, 425.
Dimethyl 28-(β-D-Glucopyranosyloxy)-2,3-secolup-20(29)-ene-2,3-dioate (47a)
Deacetylation of glucoside acetate 46a (70 mg, 0.08 mmol) afforded amorphous glucoside 47a (50 mg,
89%), [α]D –2° (c 0.49, methanol). FAB-MS, m/z: 701 (M + Na), 679 (M + H), 619, 517, 499, 457,
439.
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Dimethyl 28-(β-D-Glucopyranosyloxy)-2,3-secolupane-2,3-dioate (47b)
1797
Deacetylation of glucoside acetate 46b (120 mg, 0.14 mmol) afforded amorphous glucoside 47b (70 mg,
73%), [α]D –22° (c 0.44, methanol). FAB-MS, m/z: 703 (M + Na), 681 (M + H), 621, 519, 501, 459,
441.
The authors are indebted to the Grant Agency of the Czech Republic for the financial support (Grant
No. 203/93/2468), to Dr. S. Hilgard for the measurement of IR spectra and to the staff of the Central
Laboratories of Mass Spectrometry (Dr. K. Ubik, Head), Institute of Organic Chemistry and
Biochemistry, Academy of Sciences of the Czech Republic, for the FAB-MS measurements.
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