Synthesis of lupane triterpenoids with triphenylphosphonium substituents and studies of their antitumor activity

Article abstract of DOI:10.1007/s11172-013-0028-y

New derivatives of lupane triterpenoids, viz., 20,29-dihydrobetulinic and 3-epi-20,29-dihydrobetulinic acid derivatives containing triphenylphosphonium fragments as substituents were synthesized. These compound considerably exceed betulinic acid in antitu

Full text of DOI:10.1007/s11172-013-0028-y

1
88
Russian Chemical Bulletin, International Edition, Vol. 62, No. 1, pp. 188—198, January, 2013
Synthesis of lupane triterpenoids with triphenylphosphonium
substituents and studies of their antitumor activity
A. Yu. Spivak,a D. A. Nedopekina, E. R. Shakurova, R. R. Khalitova, R. R. Gubaidullin, V. N. Odinokov,
a
a
a
a
a
a
b
b
b
b
b
U. M. Dzhemilev, Yu. P. Bel´skii, N. V. Bel´skaya, S. A. Stankevich, E. V. Korotkaya, and V. A. Khazanov
aInstitute of Petrochemistry and Catalysis, Russian Academy of Sciences,
1
41 prosp. Oktyabrya, 450075 Ufa, Russian Federation.
Fax: +7 (347) 284 2750. Eꢀmail: ink@anrb.ru
b
Innovative Pharmacology Research (IPHAR),
8
/8 prosp. Akademicheskii, 634055 Tomsk, Russian Federation.
Fax: +7 (382) 224 8721. Eꢀmail: mail@iphar.ru
New derivatives of lupane triterpenoids, viz., 20,29ꢀdihydrobetulinic and 3ꢀepiꢀ20,29ꢀdiꢀ
hydrobetulinic acid derivatives containing triphenylphosphonium fragments as substituents
were synthesized. These compound considerably exceed betulinic acid in antitumor activity.
Key words: lupane triterpenoids, betulinic acid, triphenylphosphonium compounds, antiꢀ
cancer compounds.
Native pentacyclic lupane triterpenoids (betulin, betuꢀ
literature data on the synthesis of phosphonium salts of
lupane pentacyclic triterpenoids and on the studies of their
cytotoxic effect toward cancer cells.
linic acid) and their synthetic derivatives are an important
class of biologically active compounds with a wide range
of biological and pharmacological effects. Lupane triꢀ
terpenoids are of special interest due to their antitumor
and antiviral properties.1 In 1995, betulinic acid was
discovered to exhibit high cytotoxicity toward human melꢀ
anoma by the induction of the cancer cells apoptosis
Results and Discussion
—3
Synthesis. The present studies are devoted to the deꢀ
velopment of efficient methods for the conversion of diꢀ
hydrobetulinic (1) and 3ꢀepiꢀdihydrobetulinic acids (2) to
the earlier unknown triphenylphosphonium salts 3—8 and
to the in vitro studies of cytotoxic effect of the latter on
the Ehrlich carcinoma and mastocytoma Pꢀ815 cancer
cell lines.
–
1 4,5
(
ED50 1.1—4.8 g mL ). Later, this natural compound
was reported to display antitumor activity toward other
types of malignant cells.6 Betulinic acid and its certain
synthetic derivatives belong to a group of mitocanes, i.e.,
anticancer compounds whose biological targets are mitoꢀ
,7
8
chondria. This compound promotes accumulation of acꢀ
The key intermediates in the synthesis of the target
phosphonium salts 3—8 (Scheme 1), i.e. 2ꢀꢀallylꢀsubstiꢀ
tuted methylꢀ and benzyldihydrobetulonates 12 and 13,
were obtained based on betulonic acid 9 available from
tive oxygenꢀcontaining particles in the cancer cell mitoꢀ
chondria, that leads to the increase in the mitochondria
membrane penetrability and induction of cell apoptosis
7
,9,10
17
independent of their p53ꢀstatus.
betulin, which was transformed into compounds 10 and
1
8,19
In the series of mitochondrionꢀtargeted antitumor
compounds, the most promising results were obtained
when small positively charged molecules were used, in
particular, the molecules conjugated with a lipophilic
11 using typical procedures.
Dihydrobetulonates 12 and 13 were obtained by the
reaction of deprotonated ketones 10 and 11 with allyl
bromide in 1,2ꢀdimethoxyethane under the kinetic conꢀ
trol conditions according to the method reported by us
1
1,12
membraneꢀpenetrating triphenylphosphonium cation.
From the literature data,1
3—16
it follows that compounds
earlier (see Scheme 1). The reduction of the keto group
20
containing triphenylphosphonium cation are promising
ionic molecules for the targeted delivery of neutral biologꢀ
ically active compounds into cancer cell mitochondria,
including antitumor agents possessing prooxidant properꢀ
ties and destabilizing mitochondrion membranes. Betuꢀ
linic acid and its semiꢀsynthetic derivatives, undoubtedly,
belong to such compounds. At the same time, there are no
in compounds 12 and 13 with NaBH or Lꢀselectride gives
4
their 2ꢀhydroxy derivatives, which are necessary for subseꢀ
quent transformations into triphenylphosphonium salts
(Scheme 2). The studies of this reaction allowed us to
reveal a considerable influence of the C(2)ꢀallylic fragꢀ
ment on stereochemistry of the reduction process of 3ꢀketo
group in ring A. Thus, in contrast to the highly stereoseꢀ
Published in Russian in Izvestiya Akademii Nauk. Seriya Khimicheskaya, No. 1, pp. 0189—0199, January, 2013.
066ꢀ5285/13/6201ꢀ188 © 2013 Springer Science+Business Media, Inc.
1

Lupane triphenylphosphonium compounds
Russ.Chem.Bull., Int.Ed., Vol. 62, No. 1, January, 2013
189
1
5 indicates the axial position of proton H(3) and, conseꢀ
quently, ꢀorientation of the OHꢀgroup (in ꢀepimer 16,
3
the spinꢀspin coupling constant J
= 1.5—2 Hz).
H(2),H(3)
The stereoselectivity of the reaction was considerably inꢀ
2
2
creased by using NaBH modified with CeCl •7H O. In
4
3
2
this case, 3ꢀepimers 14 and 15 were obtained with
2
2,23
9
5—96% selectivity. The authors of the work
explain
the stereospecificity of the reduction of ketones with the
reagent NaBH —CeCl •7H O by the formation of a complex
4
3
2
of the cerium ion with the carbonyl oxygen atom, that
promotes the "axial" attack by the hydride anion on the
ketone with subsequent formation of an equatorial alcohol.
The influence of the allylic substituent in compounds
1
2 and 13 was found to exist also when a bulky reducing
agent lithium tri(secꢀbutyl)borohydride (Lꢀselectride)
was used. The reaction of dihydrobetulonate 12 with
Lꢀselectride led to 3ꢀalcohol 16 with high selectivity
(
3ꢀOH : 3ꢀOH = 96 : 4), that considerably differed from
the results of reduction of betulonic acid with Lꢀselecꢀ
2
4
tride. The obtained alcohols 14, 16 and their acetates
7—19 were converted into phosphonium salts 3—8 in
good yields (see Scheme 2). To achieve this, the double
bond in compounds 14 and 16—19 was hydroborated, the
primary alcohols 20, 21, and 24 were converted to iodides
1
Compound
R1
OH
H
OAc
H
OH
H
OAc
OAc
R2
H
OH
H
OAc
H
OH
H
R3
—
—
OMe
OMe
OMe
OMe
OH
Hal
—
—
Br
Br
I
I
Br
I
1
2
3
4
5
6
7
8
2
5—27 upon the action of iodine in the presence of imidꢀ
azole and triphenylphosphine, whereas alcohols 22—24
were converted to bromides 31—33 through the step of the
corresponding mesylates 28—30. The reaction of halide
H
OBn
25—27, 31, 32, and 34 with excess of triphenylphosphine
lective transformation of betulonic acid into betulinic acid
3ꢀOH : 3ꢀOH = 94 : 6),17,21 the reaction of comꢀ
pounds 12 and 13 with NaBH in the MeOH—CHCl or
in refluxing toluene gave the target triphenylphosphonium
salts 3—8 (see Scheme 2).
(
The structures of all the compounds obtained were
4
3
1
13
31
MeOH—THF solvent systems led to a mixture of epimers
4 and 15 with a slight predominance of the 3ꢀepimer
3ꢀOH : 3ꢀOH = 68 : 32). The spinꢀspin coupling conꢀ
confirmed by 1D ( H, C, APT, P), 2D homoꢀ (COSY,
NOESY) and heteroꢀNMRꢀexperiments (HSQC, HMBC).
1
13
(
The C NMR spectra of compounds 3—34 are given in
Tables 1—3. In the P NMR spectra of salts 3—8, the
3
31
stant ( J
= 10 Hz) of proton H(3) with the axial
H(2),H(3)
proton H(2) (the axial position of the latter was estabꢀ
lished earlier ) in the H NMR spectra of compounds 14 and
signals for the phosphorus atom were observed in the reꢀ
gion  23.37—24.34 indicative for phosphonium salts.
20
1
25
Scheme 1
R = Me (10,12), Bn (11,13)
Reagents and conditions: a. H , Pd/C, MeOH—THF (50 : 50), 20 C, 94%; b. CH N , Et O, 20 C, yield 89%, or BnCl, DMF,
2
2
2
2
K CO , 55 C, yield 92%; c. KN(SiMe ) , Et B, C H Br, dimethoxyethane, 20 C, Ar, 77—79%.
2
3
3 2
3
3
5

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Russ.Chem.Bull., Int.Ed., Vol. 62, No. 1, January, 2013
Spivak et al.
Scheme 2
R = Me (12), OBn (13); R = OH, R = H, R = Me (14, 20, 25); R1 = OH, R2 = H, R3 = Bn (15); R1 = H, R2 = OH,
1
2
3
3
= Me (16, 21, 26); R = OAc, R = H, R = Me (17, 22, 28, 31); R = OAc, R2 = H, R3 = Bn (18, 24, 27, 30, 33);
= H, R = OAc, R = Me (19, 23, 29, 32); R = OAc, R = H, R3 = H (34)
1
2
3
1
R
R
1
2
3
1
2
Reagents and conditions: a. NaBH , CeCl •7H O, MeOH—THF, –30 C20 C, Ar; b. LꢀSelectride, THF, –78 C20 C, Ar;
4
3
2
c. Ac O, Py, DMAP, 20 C; d. BH •THF, THF, 20 C, Ar; e. I , PPh , imidazole, THF, 0 C; f. MsCl, Py, CH Cl , DMAP, 20 C;
2
3
2
3
2
2
g. LiBr, Me CO, reflux, Ar; h. Pd/C, Et O; i. PPh , CH Ph, reflux, Ar.
2
2
3
3

Lupane triphenylphosphonium compounds
Russ.Chem.Bull., Int.Ed., Vol. 62, No. 1, January, 2013
191
Table 1. 13C NMR spectra (, J/Hz) of compounds 3—8 and 12—14
Atom or
group
3
4
5
6
7
8
12
13
14
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
1
44.18
34.22
82.84
38.82
55.17
18.30
32.51
40.71
50.22
37.26
20.96
23.00
38.01
42.53
29.58
32.03
56.96
48.90
44.27
28.15
29.80
37.10
26.82
16.94
17.08
15.95
14.71
176.83
22.75
14.53
32.97
JС,Р = 15)
19.09
40.81
32.24
78.42
37.34
49.93
17.96
34.15
41.42
49.93
37.67
20.69
22.79
38.07
42.62
29.65
32.14
57.00
48.89
44.22
27.75
29.65
37.65
26.75
21.28
16.77
16.00
14.84
176.95
23.00
14.65
33.42
44.22
34.34
82.61
39.29
55.25
18.51
34.52
40.69
50.14
37.28
20.95
23.01
38.12
42.55
29.63
32.05
56.99
48.92
44.21
28.52
29.77
37.13
26.94
16.91
16.93
15.96
14.71
176.89
22.76
15.01
34.54
44.36
32.09
77.43
37.28
48.86
19.33
34.91
40.66
51.17
36.76
21.27
22.87
38.13
42.53
29.65
32.02
57.02
48.86
44.18
33.45
29.74
36.76
26.92
22.76
16.83
16.38
14.72
176.88
22.99
14.59
33.10
44.14
34.23
82.92
38.85
55.16
18.30
32.52
40.73
50.19
37.40
20.93
23.02
38.11
42.58
29.61
32.07
56.77
48.72
44.30
28.17
29.79
37.13
26.81
16.94
17.08
16.07
14.71
181.81
22.75
14.52
33.03
44.16
34.20
82.80
38.84
55.15
18.30
32.47
40.69
50.23
37.23
20.97
23.01
37.93
42.54
29.47
31.97
56.93
48.89
44.27
28.15
29.80
37.09
26.84
15.82
17.09
16.98
14.70
175.97
22.73
14.10
33.10
46.86
41.50
216.97 217.19
46.86
41.49
44.14
35.34
82.61
39.12
55.49
18.49
34.36
40.69
50.32
37.89
20.90
22.97
38.09
42.53
29.63
32.06
56.95
48.93
45.03
28.43
29.72
37.17
26.94
16.22
16.82
15.97
14.69
48.30
57.95
19.34
34.16
40.76
49.86
37.35
21.17
25.27
38.09
42.65
29.63
32.06
56.95
48.92
44.19
26.84
29.75
37.29
22.96
21.74
16.12
16.04
14.67
48.30
57.25
19.34
34.14
40.74
49.87
37.33
21.16
25.28
38.01
42.66
29.52
32.00
56.92
48.91
44.17
26.87
29.76
37.27
22.98
21.75
16.06
15.99
14.67
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
´
176.70 175.97 176.82
22.76
14.55
34.55
22.74
14.52
34.56
22.74
14.64
37.29
(1
(1
(1
( J
1
= 16)
( J
1
= 16)
( J
1
= 16)
( J
1
= 15)
( J
1
= 15)
С,Р
С,Р
С,Р
С,Р
С,Р
2
3
´
´
20.12
= 4)
20.38
= 4)
19.95
= 4)
19.08
= 3)
19.02
= 3)
136.78 136.93 137.52
116.29 116.18 116.05
1
1
1
1
1
JС,Р = 4)
22.98
JС,Р = 66)
51.17
171.52
21.41
—
( J
( J
( J
( J
( J
С,Р
С,Р
С,Р
С,Р
С,Р
23.06
23.48
22.51
22.99
23.35
1
1
1
1
1
( J
= 68)
( J
= 48)
( J
= 68)
( J
= 50)
( J
= 49)
С,Р
С,Р
С,Р
С,Р
С,Р
ОМе
МеСО
МеСО
ОCH Ph
Ph
51.15
171.39
21.61
51.16
51.89
—
171.55
21.38
—
171.51
21.40
65.61
118.18 (d,
51.18
—
—
—
—
—
—
—
65.63
128.02,
128.27,
128.47,
136.58
51.15
—
—
—
—
—
—
—
—
—
—
—
—
2
118.47 (d,
1J
= 85);
118.43 (d,
118.50 (d,
1J
= 85);
118.40 (d,
1J
= 85);
118.44 (d,
1J
= 85);
1J
= 84);
1J
= 85);
С,Р
С,Р
С,Р
С,Р
С,Р
С,Р
1
1J
1
30.50 (d,
130.44 (d,
1J
= 12);
130.50 (d,
1J
= 13);
130.60 (d,
1J
= 13);
130.50 (d,
3J
= 12);
130.50 (d,
3J
= 12);
= 12);
С,Р
С,Р
С,Р
С,Р
С,Р
С,Р
33.74 (d,
132.08 (d,
1J
= 10);
133.76 (d,
1J
= 10);
132.71 (d,
1J
= 10);
133.74 (d,
JС,Р = 9);
133.69 (d,
2J
= 10);
2J
= 10);
2
С,Р
С,Р
С,Р
С,Р
С,Р
1
34.95 (br.s) 134.91 (br.s) 135.05 (br.s) 135.15 (br.s) 134.97 (br.s)
128.01,
1
1
1
28.23,
28.44,
35.09,
1
36.52
Pharmacological studies. Cytotoxic effect of phosꢀ
phonium salts on the Ehrlich carcinoma and mastoꢀ
cytoma Pꢀ815 cancer cell lines was studied in vitro. Betuꢀ
linic acid was a comparison compound. The mastoꢀ
cytoma Pꢀ815 and Ehrlich carcinoma cell lines were
used for the study of antitumor activity. Ehrlich carcinoꢀ

1
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Russ.Chem.Bull., Int.Ed., Vol. 62, No. 1, January, 2013
Spivak et al.
Table 2. 13C NMR spectra (, J/Hz) of compounds 15—24
Atom or
group
15
16
17
18
19
20
21
22
23
24
1
2
3
4
5
6
7
8
9
44.18
35.40
82.76
39.13
55.48
18.49
34.36
40.71
50.37
37.93
20.92
22.98
38.06
42.59
29.55
32.05
56.97
48.95
45.08
28.41
29.76
37.20
26.98
15.86
16.84
16.20
14.60
176.80
22.75
14.68
37.30
137.56
116.11
—
39.99
32.88
78.37
38.08
48.96
18.24
34.30
40.91
50.12
37.87
20.78
22.97
38.30
42.63
29.63
32.09
57.01
48.96
44.21
28.58
29.73
37.32
26.92
21.87
16.74
16.01
14.71
176.88
22.77
14.69
37.74
137.55
115.74
51.13
44.19
34.33
83.72
38.89
55.35
18.37
33.30
40.75
50.29
37.31
20.93
22.96
38.09
42.56
29.63
32.08
56.98
48.94
44.96
28.18
29.74
37.13
26.90
16.93
17.08
15.99
14.68
176.76
22.76
14.56
37.27
136.86
116.00
51.15
44.18
34.32
83.73
38.89
55.35
18.37
33.31
40.74
50.31
37.27
20.93
22.98
38.02
42.57
29.53
32.02
56.96
48.94
44.97
28.19
29.75
37.27
26.93
15.86
17.08
16.94
14.67
176.85
22.74
14.53
37.12
136.87
116.00
—
40.47
32.65
77.36
37.96
50.11
18.23
34.22
40.74
50.15
37.66
20.75
22.96
38.08
42.64
29.68
32.11
56.99
48.95
44.19
27.59
29.74
37.31
26.92
21.06
16.70
16.02
14.85
176.32
22.76
14.56
37.47
136.85
116.02
51.15
44.18
34.39
82.62
39.15
55.47
18.52
34.54
40.71
50.37
37.31
20.94
22.98
38.07
42.55
29.63
32.09
56.98
48.93
45.20
28.51
29.73
37.13
26.94
16.29
16.84
15.97
14.69
176.92
22.76
14.63
29.63
28.68
63.61
51.17
40.69
32.65
78.45
38.08
49.01
18.21
34.30
40.91
50.10
37.68
20.78
22.98
38.25
42.63
29.63
32.10
57.02
48.95
44.22
28.65
29.73
37.32
26.90
20.78
16.78
16.01
14.72
176.94
22.77
14.69
29.15
30.17
63.22
51.15
44.20
34.32
83.84
38.83
55.28
18.37
33.10
40.76
50.34
37.30
20.97
22.98
38.09
42.57
29.63
32.08
56.99
48.93
44.74
28.24
29.74
37.09
26.87
16.92
17.08
15.99
14.68
176.64
22.76
14.57
29.65
28.11
63.15
51.17
40.89
32.25
79.95
37.83
50.12
18.02
34.22
41.14
50.12
37.67
20.75
22.98
38.08
42.65
29.68
32.12
57.00
48.94
44.21
27.70
29.74
37.32
26.90
21.07
16.76
16.04
14.85
177.31
22.77
14.66
28.80
30.35
63.00
51.17
44.19
34.32
83.85
38.82
55.28
18.37
33.10
40.75
50.36
37.28
20.97
22.98
38.03
42.58
29.53
32.03
56.97
48.93
44.75
28.24
29.75
37.08
26.90
15.87
17.08
16.93
14.67
176.70
22.75
14.53
29.65
28.11
63.16
—
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
1
2
3
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
´
´
´
ОМе
МеСО
МеСО
Me—S
—
—
—
—
—
—
171.30
21.10
—
—
—
171.20
21.10
—
171.20
21.60
—
—
—
—
—
—
—
—
—
—
—
—
—
171.30
21.11
—
—
—
171.30
21.59
—
—
—
172.43
21.10
—
—
—
ОCH Ph
65.61
128.00,
28.25,
28.46,
36.63
65.60
65.60
2
Ph
127.99,
128.25,
128.45,
136.87
127.99,
128.25,
128.45,
138.40
1
1
1
4
–1
ma was supported in vivo in the BALB/c mice line
by intraperitoneal transplantation method. Mastocytoma
Pꢀ815 was supported in vitro. The animals were kept in
compliance with the rules of laboratory practice (GLP,
Good Laboratory Practice) and the order of the Minisꢀ
try of Health of Russian Federation No. 267 dated
by 19.06.2003 "On approval of the rules of laboratory
practice".
tion of 2•10 mL at 37 C in CO atmosphere (5%) and
2
absolute humidity over 48 h in the presence of compounds
under study in different dilutions. Four hours before stopꢀ
ping the incubation, 1ꢀ(4,5ꢀdimethylthiazolꢀ2ꢀyl)ꢀ2,5ꢀ
diphenyltetrazolium bromide (MTT, Merck Biosciences)
was added into the pits. After the incubation was stopped,
the supernatant was removed, the precipitate was dissolved
in DMSO, and absorption was measured using light soluꢀ
tions with the wavelength of 530 nm on a Uniplan spectroꢀ
photometer (PIKON Ltd.).
2
6
To determine antitumor activity, the tumor cells were
cultured in 96ꢀpit roundꢀbottom plates in the concentraꢀ

Lupane triphenylphosphonium compounds
Russ.Chem.Bull., Int.Ed., Vol. 62, No. 1, January, 2013
193
Table 3. 13C NMR spectra (, J/Hz) of compounds 25—34
Atom or
group
25
26
27
28
29
30
31
32
33
34
1
2
3
4
5
6
7
8
9
44.20
34.79
82.66
39.16
55.38
18.52
34.36
40.73
50.37
37.32
20.96
22.97
38.13
42.58
29.64
32.11
56.98
48.95
45.07
28.43
29.75
37.19
26.93
16.14
16.85
15.99
14.69
176.88
22.76
14.64
29.63
30.97
7.50
40.32
32.10
78.53
38.06
48.95
18.26
34.29
40.91
50.10
37.69
20.79
22.78
38.31
42.63
29.63
31.37
57.01
48.95
44.21
28.58
29.74
37.32
26.89
20.79
16.79
16.02
14.72
176.89
22.78
14.71
34.18
32.09
7.28
44.19
34.31
83.56
38.85
55.23
18.36
32.77
40.76
50.34
37.28
20.98
22.98
38.02
42.59
29.53
32.03
56.96
48.93
44.94
28.24
29.76
37.13
26.91
15.87
17.07
16.95
14.68
176.00
22.75
14.54
30.73
33.25
7.15
44.20
34.30
83.46
38.85
55.26
18.36
32.90
40.77
50.33
37.30
20.98
22.97
38.08
42.58
29.62
32.08
56.97
48.92
44.65
28.21
29.74
37.13
26.86
16.92
17.05
15.99
14.68
176.86
22.76
14.56
27.96
26.18
70.27
51.16
171.29
21.04
37.40
—
40.87
32.11
79.36
37.82
50.05
17.99
34.18
41.10
50.09
37.67
20.75
22.97
38.05
42.64
29.66
31.98
56.98
48.92
44.19
27.67
29.73
37.31
26.86
21.01
16.75
16.02
14.84
176.85
22.76
14.66
28.57
26.71
70.26
51.16
171.07
21.58
37.35
—
44.19
34.29
83.47
38.85
55.26
18.36
32.90
40.75
50.35
37.27
20.97
22.97
38.01
42.59
29.52
32.02
56.95
48.92
44.66
28.21
29.75
37.11
26.89
16.93
17.05
15.87
14.67
175.99
22.74
14.53
27.96
26.18
70.26
—
44.20
34.31
83.56
38.85
55.25
18.36
32.89
40.77
50.32
37.30
20.98
22.97
38.09
42.57
29.63
32.08
56.98
48.93
44.86
28.23
29.75
37.13
26.88
16.93
17.07
16.00
14.69
176.86
22.76
14.56
29.98
30.87
34.02
51.17
171.30
21.09
—
40.90
32.13
79.64
37.87
50.08
18.00
34.22
41.09
50.08
37.68
20.77
22.97
38.09
42.66
29.69
31.82
57.00
48.96
44.22
27.65
29.75
37.32
26.89
21.04
16.77
16.04
14.66
177.55
22.78
14.85
30.27
31.38
34.13
51.16
171.02
21.59
—
44.19
34.30
83.57
38.85
55.24
18.36
32.89
40.75
50.34
37.27
20.98
22.97
38.02
42.58
29.52
32.02
56.96
48.93
44.87
28.23
29.76
37.12
26.90
16.94
17.06
15.87
14.67
176.00
22.74
14.53
29.99
30.87
34.00
—
44.18
34.30
83.58
38.86
55.23
18.34
32.91
40.77
50.27
37.41
20.93
22.97
38.25
42.61
29.64
32.06
56.83
48.75
44.86
28.24
29.73
37.16
26.83
16.91
17.05
16.10
14.67
182.56
22.73
14.56
30.00
30.88
34.00
—
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
1
2
3
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
´
´
´
ОМе
51.17
—
—
—
—
51.15
—
—
—
—
—
МеСО
МеСО
Me—S
ОCH Ph
Ph
171.26
21.14
—
65.61
127.99,
171.27
21.04
37.41
65.60
127.99,
128.25,
128.45,
136.59
171.27
21.08
—
65.60
127.99,
128.25,
128.45,
136.60
171.32
21.08
—
—
—
—
—
—
—
2
—
—
—
—
1
1
1
28.25,
28.46,
36.60
–
1
IC /mol L
As it follows from the experimental results (Table 4,
Fig. 1), all the salts under study 3, 4, 7, and 8 considerably (by
5
0
2
6
5
4
3
2
1
0
3
4—40 times) exceeded betulinic acid in antitumor activity.
In conclusion, we developed an efficient approach to
0
0
0
0
0
1
the synthesis of earlier unknown phosphoniumꢀcontainꢀ
ing lupane triterpenoids possessing high cytotoxic activiꢀ
ty. The availability of the starting plantꢀoriginated metaꢀ
bolites (betulin, betulonic and betulinic acids) and high
yields in all the steps of the synthesis make this approach
promising for the preparation of a large group of pentaꢀ
cyclic triterpene conjugates with phosphonium cations as
potential antitumor agents.
3
4
7
8
BA Agent
Fig. 1. Cytotoxic effect of betulinic acid (BA) and salts 3, 4, 7,
and 8 on mastocytoma Pꢀ815 (1) and Ehrlich carcinoma (2)
tumor cells.

1
94
Russ.Chem.Bull., Int.Ed., Vol. 62, No. 1, January, 2013
Spivak et al.
Table 4. Cytotoxic effect of phosphonium salts 3, 4, 7, 8 and betulinic acid (BA) on the Pꢀ815 (in the numerator)
and Ehrlich tumor cells (in the denominator) (% of viable cells) (X±SE)a
Concentration of
3
4
7
8
BA
—
compound/g mL–1
—b
—
—
100.0±2.2
—
1
00.0±2.4
0
1
1
2
5
.1
98.6±4.2
96.1±3.8
95.1±4.9
97.5±4.5
98.6±2.6
97.6±2.3
98.1±3.8
89.1±6.1
99.8±4.8
89.3±4.1
93.3±4.4
102.8±6.0
96.8±3.1
9
5.5±1.2
c,d
c,d
c,d
59.7±3.7
3.4±5.02
–6.8±2.9
3.9±4.62
53.8±5.0
71.7±3.62
–3.5±1.6
60.0±4.5
73.4±2.32
–11.2±3.7
16.0±2.22
c,d
c,d
c,d
c,d
7
c,d
c,d
c,d
c
0
5
0
13.4±4.4
10.8±1.12
62.8±1.1
57.3±4.2
34.3±5.0
50.4±2.4
30.5±2.9
40.5±2.6
c,d
c,d
c,d
c,d
c
–
–1.3±2.02
c,d
c,d
c,d
c,d
c
7.8±6.6
3.8±1.0
–2.8±3.2
2.2±4.2
c,d
c,d
c,d
c,d
c
7
.2±4.82
–3.8±2.22
5.4±2.62
1.2±2.32
c,d
c,d
c,d
c,d
c
8.3±2.8
2.1±7.92c,d
1.20
5.0±1.6
0.2±1.92
1.15
7.2±3.7
–5.2±5.02
1.10
4.6±2.2
0.1±4.02
4.79
c,d
c,d
c,d
c
–
IC50e/mol L–1
41.00
54.00
1
.37
1.35
2.30
4.70
a
X is the average value of the experimental data, SE is the standard error.
Control.
The differences with the control are reliable, p < 0.05.
The differences with the corresponding concentration of betulinic acid are reliable, p < 0.05.
IC50 is the concentration causing a halfꢀmaximal inhibition of viable cells.
b
c
d
e
Experimental
and this was stirred for 1 h, followed by addition of a solution of
allyl bromide (0.1 g, 0.84 mmol) in DME (1 mL). The reaction
mixture was stirred for 4 h (TLC monitoring), then neutralized
IR spectra were recorded on a Specord IRꢀ75 spectrometer
with 3 M HCl, diluted with H O (1.5 mL), and extracted with
EtOAc (4×10 mL). The combined extracts were dried with
2
1
13
31
for neat samples or for solutions in CHCl . H, C, and
P
3
NMR spectra were recorded on a Bruker Avanceꢀ400 spectroꢀ
meter ( H, 400.13 MHz; C, 100.62 MHz; P, 161.98 MHz)
MgSO . The residue was concentrated and subjected to chromatoꢀ
4
1
13
31
graphy on a column with SiO (eluent hexane—EtOAc) to obꢀ
2
using Me Si as an internal standard and CDCl3 as a solvent.
tain compound 12 (0.15 g, 65%) as white crystals, m.p. 84—86 C
4
Mass spectra were recorded on a BrukerꢀAutoflex III instruꢀ
ment in the MALDIꢀTOF regime with recording positive ions
and using 2,5ꢀdihydroxybenzoic and ꢀcyanoꢀ4ꢀhydroxycinꢀ
namic acids as matrices. Optical rotation was measured on
a Perkin—Elmerꢀ141 polarimeter. Specific rotation is given in
(EtOH), []D20 –36.0 (c 0.35, CHCl ). Found (%): C, 80.03;
3
H, 10.61. C H O . Calculated (%): C, 79.95; H, 10.66. IR,
3
4
54
3
–
1
1
/cm : 1728 (C=O). H NMR, : 0.76, 0.87 (both d, 3 H each,
H(29), H(30), J = 6 Hz); 0.91 (m, 1 H, H eq(1)); 0.95, 0.98,
a
1.06, 1.07, 1.11 (all s, 3 H each, H(23)—H(27)); 1.13—2.25
–
1
–1
deg mL g dm , the concentration of solutions is given in
(m, 21 H, CH, CH in pentacyclic skeleton and 1 H, H(20));
2
–
1
2
3
g (100 mL) . Elemental analysis was performed on a Karlo Erba
106 analyzer. Sorbfil plates (Sorbpolimer, Krasnodar, Russia)
were used for TLC, visualizing with anisaldehyde. Silica gel L
50—160 m, KSKG) was used for column chromatography.
The following reagents were used in the work: BH •THF
1.98 (dt, 1 H, H(1´), J = 15 Hz, J = 7 Hz); 2.10 (dd, 1 H,
b
2
3
1
H
(1), J = 13 Hz, J = 6 Hz); 2.55 (m, 1 H, H(1´)); 2.72
ax
(m, 1 H, H(2)); 3.67 (s, 3 H, OMe); 5.00—5.05 (m, 2 H,
+
(
H(3´)); 5.76 (m, 1 H, H(2´)). MS, m/z: 533.4 [M + Na] , 549.4
+
+
[M + K] . C H O . Calculated, m/z: 533.4 [M + Na] ,
3
34 54
3
+
(
(
(
1 M solution in THF), BEt (1 M solution in THF), KN(SiMe3)2
549.4 [M + K] .
3
1 M solution in THF), Lꢀselectride, CeCl •7H O, DME
Benzyl 2ꢀallylꢀ3ꢀoxoꢀ20,29ꢀdihydrobetulonate (13) was
obtained similarly to compound 12. The yield was 67%. White
crystals, m.p. 132—134 C (EtOH), []D20 –21.9 (c 0.70,
CHCl ). Found (%): C, 81.93; H, 9.91. C H O . Calculatꢀ
3
2
dimethoxyethane), allyl bromide (Aldrich). Hexane, THF, and
DME were refluxed and distilled over sodium. Compounds 12
and 13 were synthesized according to the method described earꢀ
3
40 58
3
2
0
–1
1
lier. Betulonic acid was obtained from betulin according to the
known procedures.17,27 The carboxylic function in compound
ed (%): C, 81.86; H, 9.96. IR, /cm : 1726 (C=O). H NMR,
: 0.75, 0.86 (both d, 3 H each, H(29), H(30), J = 7 Hz); 0.81,
0.93, 1.06, 1.07, 1.09 (all s, 3 H each, H(23)—H(27)); 0.90
3
3 was deblocked according to the typical procedure.19 Comꢀ
a
pounds 14—16 were acetylated according to the standard proceꢀ
dure. Acetates 17—19 were purified by chromatography.
Methyl 2ꢀallylꢀ3ꢀoxoꢀ20,29ꢀdihydrobetulonate (12). A 1 M
solution of KN(SiMe ) in THF (0.55 mL, 0.55 mmol) was addꢀ
(m, 1 H, H (1)); 1.12—2.27 (m, 21 H, CH, CH in pentacyclic
eq
2
2
skeleton and 1 H, H(20)); 1.97 (dt, 1 H, H(1´), J = 15 Hz,
3
J = 7 Hz); 2.10 (dd, 1 H, Hb (1), J = 13 Hz, J = 6 Hz); 2.57
2
3
ax
(m, 1 H, H(1´)); 2.71 (m, 1 H, H(2)); 4.99—5.05 (m, 2 H,
3
2
ed to a solution of compound 10 (0.42 mmol) in DME (2.50 mL)
at ~20 C under argon with stirring. After 15 min, a 1 M solution
H(3´)); 5.13 (m, 2 H, OCH Ph); 5.78 (m, 1 H, H(2´)); 7.28—7.39
2
+
(m, 5 H, Ph). MS, m/z: 609.4 [M + Na] . C H O . Calculatꢀ
4
0
58
3
+
of Et B in THF (0.55 mL, 0.55 mmol) was added to the mixture
ed, m/z: 609.4 [M + Na] .
3

Lupane triphenylphosphonium compounds
Russ.Chem.Bull., Int.Ed., Vol. 62, No. 1, January, 2013
195
Methyl 2ꢀallylꢀ3ꢀhydroxyꢀ20,29ꢀdihydrobetulinate (14).
Methyl 3ꢀacetoxyꢀ2ꢀallylꢀ20,29ꢀdihydrobetulinate (17).
The yield was 95%. White crystals, m.p. 208—210 C (EtOH),
A solution of CeCl •7H O (0.15 g, 0.4 mmol) in a mixture of
3
2
THF—methanol (1 : 1, 1 mL) was added dropwise to a solution
of compound 12 (0.3 mmol) in a mixture of THF—methanol
[]D20 –29.6 (c 1.15, CHCl ). Found (%): C, 77.98; H, 10.49.
3
–
1
C H O . Calculated (%): C, 77.93; H, 10.54. IR, /cm : 1734
36 58 4
1
a
(
0
1 : 2, 7.5 mL) cooled to –30 C (Ar). Then, NaBH (0.023 g,
(C=O). H NMR, : 0.63 (t, 1 H, H eq(1), J = 12 Hz); 0.75, 0.85
(both d, 3 H each, H(29), H(30), J = 7 Hz); 0.82, 0.84, 0.88,
0.92, 0.95 (all s, 3 H each, H(23)—H(27)); 1.10—2.30 (m, 22 H,
4
.6 mmol) was added in small portions over 5 min, the temperaꢀ
ture was raised to ~20 C and the mixture was stirred at this
temperature for 2 h (TLC monitoring). Then, the reaction mixꢀ
ture was neutralized with 5% aqueous HCl and extracted with
ethyl acetate (3×10 mL). The extract was washed with saturated
aq. NaHCO and water, dried with MgSO , and the solvent was
CH, CH in pentacyclic skeleton, 1 H, H(20) and 2 H, H(1´));
2
b
1.73 (m, 1 H, H ax(1)); 2.09 (s, 3 H, Me (OAc)); 3.66 (s, 3 H,
OMe); 4.47 (d, 1 H, H(3), J = 11 Hz); 4.95—5.00 (m, 2 H,
+
H(3´)); 5.68—5.75 (m, 1 H, H(2´)). MS, m/z: 577.5 [M + Na] .
3
4
+
evaporated in vacuo. The residue was purified by column chroꢀ
C H O . Calculated, m/z: 577.4 [M + Na] .
3
6
58
4
matography on SiO (eluent hexane—EtOAc) to obtain comꢀ
Benzyl 3ꢀacetoxyꢀ2ꢀallylꢀ20,29ꢀdihydrobetulinate (18).
2
pound 14(0.11 g, 74%) as white crystals, m.p. 90—92 C (EtOH),
The yield was 87%. White crystals, m.p. 74—76 C (EtOH),
2
0
20
[
]D –4.7 (c 0.45, CHCl ). Found (%): C, 79.69; H, 10.57.
[]D –25.9 (c 0.36, CHCl ). Found (%): C, 80.01; H, 9.87.
3
3
–
1
–1
C H O . Calculated (%): C, 79.63; H, 11.01. IR, /cm : 1726
(
C H O . Calculated (%): C, 79.95; H, 9.90. IR, /cm : 1732
3
4
56
3
42 62
4
1
1
a
a
C=O), 3450 (OH). H NMR, : 0.55 (t, 1 H, H eq(1), J = 12 Hz);
(C=O). H NMR, : 0.63 (t, 1 H, H eq(1), J = 12 Hz); 0.75, 0.86
(both d, 3 H each, H(29), H(30), J = 7 Hz); 0.76, 0.84, 0.85,
0.87, 0.94 (all s, 3 H each, H(23)—H(27)); 1.08—2.28 (m, 22 H,
0
0
.72, 0.83 (both d, 3 H each, H(29), H(30), J = 7 Hz); 0.76,
.83, 0.89, 0.93, 0.96 (all s, 3 H each, H(23)—H(27)); 1.13—2.21
(
1
3
m, 22 H, CH, CH in pentacyclic skeleton and 1 H, H(20));
CH, CH in pentacyclic skeleton, 1 H, H(20) and 2 H, H(1´));
1.69 (m, 1 H, H ax(1)); 2.09 (s, 3 H, Me (OAc)); 4.47 (d, 1 H,
H(3), J = 11 Hz); 4.96—5.00 (m, 2 H, H(3´)); 5.12 (m, 2 H,
2
2
.73 (m, 1 H, H (1)); 1.91 (dt, 1 H, H(1´), 2J = 15 Hz,
b
b
ax
J = 7 Hz); 2.48 (m, 1 H H(1´)); 2.85 (d, 1 H, H(3), J = 10 Hz);
3
.63 (s, 3 H, COOMe); 4.98—5.04 (m, 2 H, H(3´)); 5.81 (m, 1 H,
OCH Ph); 5.69—5.75 (m, 1 H, H(2´)); 7.31—7.37 (m, 5 H, Ph).
2
+
+
H(2´)). MS, m/z: 512.4 [M] . C H O . Calculated, m/z:
5
MS, m/z: 653.4 [M + Na] . C H O . Calculated, m/z: 653.4
3
4
56
3
42 62
4
+
+
12.4 [M] .
[M + Na] .
Benzyl 2ꢀallylꢀ3ꢀhydroxyꢀ20,29ꢀdihydrobetulinate (15) was
Methyl 3ꢀacetoxyꢀ2ꢀallylꢀ20,29ꢀdihydrobetulinate (19).
The yield was 87%. White crystals, m.p. 182—184 C (EtOH),
obtained similarly to compound 14. The yield was 72%. White
crystals, m.p. 76—78 C (EtOH), []D2 –19.3 (c 0.60, CHCl ).
0
[]D –24.2 (c 0.19, CHCl ). Found (%): C, 77.99; H, 10.47.
20
3
3
–
1
Found (%): C, 81.64; H, 10.21. C H O . Calculated (%):
C H O . Calculated (%): C, 77.93; H, 10.54. IR, /cm : 1731
(C=O). H NMR, : 0.75, 0.85 (both d, 3 H each, H(29),
H(30), J = 7 Hz); 0.82, 0.84, 0.88, 0.92, 0.99 (all s, 3 H each,
4
0
60
3
36 58 4
–1
1
1
C, 81.58; H, 10.27. IR, /cm : 1726 (C=O), 3500 (OH). H NMR,
: 0.58 (t, 1 H, H eq(1), J = 12 Hz); 0.75, 0.86 (both d, 3 H each,
a

H(29), H(30), J = 7 Hz); 0.76, 0.79, 0.84, 0.95, 0.99 (all s,
H(23)—H(27)); 1.14—2.30 (m, 24 H, CH, CH in pentacyclic
2
3
H each, H(23)—H(27)); 1.11—2.24 (m, 22 H, CH, CH in
skeleton, 1 H, H(20) and 2 H, H(1´)); 2.10 (s, 3 H, Me (OAc));
2
b
pentacyclic skeleton and 1 H, H(20)); 1.70 (m, 1 H, H ax(1));
1
H(1´)); 2.89 (d, 1 H, H(3), J = 10 Hz); 5.02—5.16 (m, 2 H,
3.66 (s, 3 H, OMe); 4.77 (br.s, 1 H, H(3)); 4.95—4.99 (m, 2 H,
2
3
+
.97 (dt, 1 H, H(1´), J = 15 Hz, J = 7 Hz); 2.48 (m, 1 H,
H(3´)); 5.77—5.85 (m, 1 H, H(2´)). MS, m/z: 577.5 [M + Na] .
+
C H O . Calculated, m/z: 577.4 [M + Na] .
3
6
58
4
H(3´)); 5.09—5.13 (m, 2 H, OCH Ph); 5.80—5.91 (m, 1 H,
Methyl 3ꢀhydroxyꢀ2ꢀ(3ꢀhydroxypropyl)ꢀ20,29ꢀdihydroꢀ
2
+
H(2´)); 7.31—7.37 (m, 5 H, Ph). MS, m/z: 611.4 [M + Na] ,
betulinate (20). A 1 M solution of BH •THF in THF (0.86 mL,
3
+
+
6
6
27.3 [M + K] . C H O . Calculated, m/z: 611.4 [M + Na] ,
0.86 mmol) was added to a stirred solution of compound 14
(0.43 mmol) in anhydrous THF (5 mL) at ~20 C (Ar). After 3 h,
the reaction mixture was cooled to 0 C, followed by a careful
dropwise addition of 10% aq. NaOH (1 mL) and 30% aq. H O
4
0
60
3
+
27.4 [M + K] .
Methyl 2ꢀallylꢀ3ꢀhydroxyꢀ20,29ꢀdihydrobetulinate (16).
A 1 M solution of Lꢀselectride in THF(2.4 mL, 2.4 mmol) was
added to a solution of compound 12 (0.41 g, 0.8 mmol) in THF
2
2
(1 mL). The reaction mixture was stirred for 1 h at ~20 C,
neutralized with 3 M HCl, and extracted with ethyl acetate
(3×20 mL). The combined organic phases were washed with
(
15 mL) cooled to –78 C (Ar) with stirring. The solution was
stirred for 2 h at ~20 C, followed by the addition of a 2 M aq.
NaOH (18 mL) and 30% aq. H O (4 mL) and stirring for another
brine and dried with MgSO , the solvent was evaporated in vacuo.
2
2
4
1
h. The mixture was concentrated to a small volume and exꢀ
The residue was purified by column chromatography on SiO2
tracted with EtOAc (4×10 mL). The organic phase was washed
(eluent hexane—EtOAc) to obtain compound 20 (0.17 g, 76%)
as white crystals, m.p. 111—113 C (EtOH), []D –40 (c 0.28,
2
0
with water, dried with MgSO , and the solvent was evaporated
4
in vacuo. The residue was purified by column chromatography
CHCl ). Found (%): C, 76.82; H, 10.12. C H O . Calculatꢀ
3
34 58
4
–
1
on SiO (eluent hexane—EtOAc) to obtain compound 16 (0.28 g,
ed (%): C, 76.93; H, 11.01. IR, /cm : 1730 (C=O), 3334 (OH).
2
2
0
1
a
6
8%) as white crystals, m.p. 84—86 C (EtOH), []D –0.54
H NMR, : 0.57 (t, 1 H, H eq(1), J = 12 Hz); 0.77, 0.75 (both d,
(
c 1.66, CHCl ). Found (%): C, 79.72; H, 10.59. C H O .
3 H each, H(29), H(30), J = 7 Hz); 0.84, 0.85, 0.92, 0.95, 0.98
(all s, 3 H each, H(23)—H(27)); 1.16—2.29 (m, 22 H, CH, CH2
in pentacyclic skeleton, 1 H, H(20), 2 H, H(1´), 2 H, H(2´));
1.70 (m, 1 H, Hbax(1)); 3.64 (br.m, 2 H, H(3´)); 3.65 (s, 3 H,
OMe); 2.84 (d, 1 H, H(3), J = 10 Hz). MS, m/z: 553.4
3
34 56
3
–
1
Calculated (%): C, 79.63; H, 11.01. IR, /cm : 1729 (C=O),
3
H(30), J = 7 Hz); 0.83, 0.85, 0.91, 0.95, 0.96 (all s, 3 H each,
H(23)—H(27)); 1.12—2.30 (m, 24 H, CH, CH in pentacyclic
skeleton, 1 H, H(20) and 2 H, H(1´)); 3.21 (br.s, 1 H, H(3));
3
H(2´)). MS, m/z: 512.4 [M] . C H O . Calculated, m/z:
5
1
450 (OH). H NMR, : 0.75, 0.86 (both d, 3 H each, H(29),
2
+
+
[M + Na] , 569.3 [M + K] . C H O . Calculated, m/z: 553.4
34 58 4
+
+
.65 (s, 3 H, COOMe); 5.00—5.07 (m, 2 H, H(3´)); 5.81 (m, 1 H,
[M + Na] , 569.4 [M + K] .
+
Methyl 3ꢀhydroxyꢀ2ꢀ(3ꢀhydroxypropyl)ꢀ20,29ꢀdihydroꢀ
betulinate (21) was obtained similarly to compound 20 from comꢀ
3
4
56
3
+
12.4 [M] .

1
96
Russ.Chem.Bull., Int.Ed., Vol. 62, No. 1, January, 2013
Spivak et al.
pound 16. The yield was 78%. White crystals, m.p. 92—94 C
crystals, m.p. 97—99 C (EtOH), [] 28 –39.3 (c 0.28, CHCl ).
D
3
(
1
1
EtOH), []D20 –24 (c 0.28, CHCl ). Found (%): C, 76.87; H,
Found (%): C, 63.85; H, 8.79; I, 19.69. C H O I. Calculatꢀ
3
3
34 57
–1
–1
0.14. C H O . Calculated (%): C, 76.93; H, 11.01. IR, /cm
726 (C=O), 3363 (OH). H NMR, : 0.76, 0.87 (both d, 3 H
:
ed (%): C, 63.74; H, 8.97; I, 19.81. IR, /cm : 757 (CH —I),
34
58
4
2
1
1
a
1727 (C=O). H NMR, : 0.58 (t, 1 H, H eq(1), J = 12 Hz); 0.75,
0.88 (both d, 3 H each, H(29), H(30), J = 7 Hz); 0.78, 0.86,
0.92, 0.96, 0.99 (all s, 3 H each, H(23)—H(27)); 1.13—2.26
each, H(29), H(30), J = 7 Hz); 0.82, 0.85, 0.89, 0.91, 0.95 (all s,
3
H each, H(23)—H(27)); 1.11—2.23 (m, 24 H, CH, CH in
2
pentacyclic skeleton, 1 H, H(20), 2 H, H(1´), 2 H, H(2´)); 3.50
t, 2 H, H(3´), J = 5 Hz); 3.66 (s, 3 H, OMe); 3.23 (s, 1 H, H(3)).
(m, 22 H, CH, CH in pentacyclic skeleton, 1 H, H(20), 2 H, H(1´),
2 H, H(2´)); 1.71 (m, 1 H, H ax(1)); 3.19 (m, 2 H, H(3´)); 3.67
(s, 3 H, OMe); 2.85 (d, 1 H, H(3), J = 12 Hz).
2
b
(
+
MS, m/z: 553.4 [M + Na] . C H O . Calculated, m/z: 553.4
3
4
58
4
+
[
M + Na] .
Methyl 3ꢀhydroxyꢀ2ꢀ(3ꢀiodopropyl)ꢀ20,29ꢀdihydrobetuliꢀ
nate (26) was obtained similarly to compound 25 from comꢀ
pound 21. The yield was 73%. White crystals, m.p. 86—88 C
Methyl 3ꢀacetoxyꢀ2ꢀ(3ꢀhydroxypropyl)ꢀ20,29ꢀdihydroꢀ
betulinate (22) was obtained similarly to compound 20 from comꢀ
pound 17. The yield was 71%. White crystals, m.p. 114—116 C
(EtOH), []D28 –28.4 (c 0.28, CHCl ). Found (%): C, 63.81; H,
3
(
EtOH), []D20 –28.9 (c 1.05, CHCl ). Found (%): C, 75.57;
8.75; I, 19.69. C H O I. Calculated (%): C, 63.74; H, 8.97;
3
34 57
3
–
1
1
H, 10.49. C H O . Calculated (%): C, 75.48; H, 10.56. IR,
I, 19.81. IR, /cm : 757 (CH —I), 1728 (C=O). H NMR,
3
6
60
5
2
–
1
1

H
/cm : 1732 (C=O), 3446 (OH). H NMR, : 0.68 (t, 1 H,
eq(1), J = 12 Hz); 0.75, 0.86 (both d, 3 H each, H(29), H(30),
: 0.76, 0.87 (both d, 3 H each, H(29), H(30), J = 7 Hz); 0.83,
0.85, 0.91, 0.96, 0.98 (all s, 3 H each, H(23)—H(27)); 1.12—2.24
a
J = 7 Hz); 0.81, 0.83, 0.88, 0.92, 0.95 (all s, 3 H each,
(m, 24 H, CH, CH in pentacyclic skeleton, 1 H, H(20), 2 H,
2
H(23)—H(27)); 1.12—2.29 (m, 22 H, CH, CH in pentacyclic
H(1´), 2 H, H(2´)); 3.19 (m, 2 H, H(3´)); 3.66 (s, 3 H, OMe);
3.21 (br.s, 1 H, H(3)).
2
skeleton, 1 H, H(20), 2 H, H(1´), 2 H, H(2´)); 1.69 (m, 1 H,
b
H
ax(1)); 2.10 (s, 3 H, Me (OAc)); 3.55 (m, 2 H, H(3´)); 3.66
Benzyl 3ꢀacetoxyꢀ2ꢀ(3ꢀiodopropyl)ꢀ20,29ꢀdihydrobetulꢀ
inate (27) was obtained similarly to compound 25 from comꢀ
pound 24. The yield was 94%. White crystals, m.p. 69—71 C
(
[
[
s, 3 H, OMe); 4.46 (d, 1 H, H(3), J = 11 Hz). MS, m/z: 595.4
+
+
M + Na] , 611.4 [M + K] . C H O . Calculated, m/z: 595.4
3
6
60
5
+
+
28
M + Na] , 611.4 [M + K] .
(EtOH), []D –13.2 (c 0.28, CHCl ). Found (%): C, 66.39;
3
Methyl 3ꢀacetoxyꢀ2ꢀ(3ꢀhydroxypropyl)ꢀ20,29ꢀdihydroꢀ
H, 8.42; I, 16.68. C H O I. Calculated (%): C, 66.48; H, 8.37;
4
2
63
4
–
1
1
betulinate (23) was obtained similarly to compound 20 from comꢀ
pound 19. The yield was 74%. White crystals, m.p. 104—106 C
I, 16.72. IR, /cm : 756 (CH —I), 1731 (C=O). H NMR,
2
a
: 0.66 (t, 1 H, H eq(1), J = 12 Hz); 0.75, 0.87 (both d, 3 H each,
(
EtOH), []D20 –20.2 (c 0.22, CHCl ). Found (%): C, 75.54;
H(29), H(30), J = 7 Hz); 0.76, 0.82, 0.83, 0.86, 0.94 (all s,
3
H, 10.51. C H O . Calculated (%): C, 75.48; H, 10.56. IR,
3 H each, H(23)—H(27)); 1.09—2.28 (m, 22 H, CH, CH in
3
6
60
5
2
–
1
1

3
/cm : 1731 (C=O), 3445 (OH). H NMR, : 0.76, 0.88 (both d,
H each, H(29), H(30), J = 7 Hz); 0.82, 0.87, 0.89, 0.92, 1.00
pentacyclic skeleton, 1 H, H(20), 2 H, H(1´), 2 H, H(2´)); 1.73
(m, 1 H, Hbax(1)); 2.13 (s, 3 H, Me (OAc)); 3.14 (t, 2 H, H(3´),
J = 7 Hz); 4.45 (d, 1 H, H(3), J = 11.2 Hz); 5.09—5.16 (m, 2 H,
(
all s, 3 H each, H(23)—H(27)); 1.10—2.26 (m, 24 H, CH, CH2
in pentacyclic skeleton, 1 H, H(20), 2 H, H(1´), 2 H, H(2´));
.10 (s, 3 H, Me (OAc)); 3.59 (t, 2 H, H(3´), J = 6 Hz); 3.66 (s, 3 H,
OCH Ph); 7.28—7.35 (m, 5 H, Ph).
2
2
Methyl 3ꢀacetoxyꢀ2ꢀ(3ꢀmesyloxypropyl)ꢀ20,29ꢀdihydroꢀ
betulinate (28). Pyridine (0.06 g, 0.77 mmol) and DMAP (0.03 g,
0.25 mmol) were added to compound 22 (0.44 mmol) dissolved
in CH Cl (2 mL) and the mixture was cooled to 0 C, followed
+
OMe); 4.77 (br.s, 1 H, H(3)). MS, m/z: 595.5 [M + Na] ,
6
6
+
+
11.4 [M + K] . C H O . Calculated, m/z: 595.4 [M + Na] ,
36 60 5
+
11.4 [M + K] .
Benzyl 3ꢀacetoxyꢀ2ꢀ(3ꢀhydroxypropyl)ꢀ20,29ꢀdihydroꢀ
2
2
by a dropwise addition of a solution of methanesulfonyl chloride
betulinate (24) was obtained similarly to compound 20 from comꢀ
(0.07 g, 0.65 mmol) in CH Cl (1 mL) and stirring for 24 h at
2
2
pound 18. The yield was 79%. White crystals, m.p. 74—76 C
(
~20 C (TLC monitoring). Then, a cold 5% aq. HCl was added
to the reaction mixture, which was extracted with ethyl acetate
(3×10 mL). The extract was washed with saturated aq. NaHSO3,
EtOH), []D20 –23.2 (c 0.46, CHCl ). Found (%): C, 77.80;
3
H, 9.89. C H O . Calculated (%): C, 77.73; H, 9.94. IR,
4
2
64
5
–1
1
a

/cm : 1730 (C=O). H NMR, : 0.65 (t, 1 H, H eq(1), J = 12 Hz);
water, dried with MgSO , and the solvent was evaporated
4
0
.75, 0.86 (both d, 3 H each, H(29), H(30), J = 7 Hz); 0.77,
in vacuo. The residue was purified by column chromatography
0
.81, 0.85, 0.87, 0.94 (all s, 3 H each, H(23)—H(27)); 1.08—2.32
on SiO (eluent hexane—EtOAc) to obtain compound 28 (0.27 g,
2
2
0
(
m, 22 H, CH, CH in pentacyclic skeleton, 1 H, H(20), 2 H,
93%) as white crystals, m.p. 98—100 C (EtOH), []D –40.1
(c 0.18, CHCl ). Found (%): C, 68.33; H, 9.55; S, 4.96. C H O S.
2
b
H(1´), 2 H, H(2´)); 1.69 (m, 1 H, H ax(1)); 2.10 (s, 3 H, Me
3
37 62
7
–
1
(
OAc)); 3.57 (m, 2 H, H(3´)); 4.46 (d, 1 H, H(3), J = 11 Hz);
Calculated (%): C, 68.27; H, 9.60; S, 4.93. IR, /cm :
1
a
5
6
6
.12 (m, 2 H, OCH Ph); 7.32—7.39 (m, 5 H, Ph). MS, m/z:
1358 (S=O), 1730 (C=O). H NMR, : 0.66 (t, 1 H, H eq(1),
J = 12 Hz); 0.76, 0.86 (both d, 3 H each, H(29), H(30), J = 7 Hz);
0.81, 0.83, 0.88, 0.92, 0.95 (all s, 3 H each, H(23)—H(27));
2
+
+
49.9 [M + H] , 671.3 [M + Na] . C H O . Calculated, m/z:
4
2
64
5
+
+
49.7 [M + H] , 671.4 [M + Na] .
Methyl 3ꢀhydroxyꢀ2ꢀ(3ꢀiodopropyl)ꢀ20,29ꢀdihydrobetulꢀ
inate (25). Triphenylphosphine (0.22 g, 0.84 mmol), imidazole
0.12 g, 1.17 mmol), and crystalline iodine (0.19 g, 0.75 mmol)
1.10—2.25 (m, 22 H, CH, CH in pentacyclic skeleton, 1 H,
2
b
H(20), 2 H, H(1´), 2 H, H(2´)); 1.69 (m, 1 H, H ax(1)); 2.11
(
(s, 3 H, Me (OAc)); 3.00 (s, 3 H, OMs); 3.66 (s, 3 H, OMe); 4.18
were added to a solution of compound 20 (0.35 mmol) in anꢀ
hydrous THF (6 mL) at 0 C (Ar) with stirring and the mixture
was stirred for 1 h at 0 C (TLC monitoring). After the reaction
reached completion, the solution was diluted with ethyl acetate
(t, 2 H, H(3´), J = 6 Hz); 4.45 (d, 1 H, H(3), J = 11 Hz). MS,
+
+
m/z: 673.3 [M + Na] , 689.3 [M + K] . C H O S. Calculatꢀ
3
7
62
7
+
+
ed, m/z: 673.4 [M + Na] , 689.4 [M + K] .
Methyl 3ꢀacetoxyꢀ2ꢀ(3ꢀmesyloxypropyl)ꢀ20,29ꢀdihydroꢀ
betulinate (29) was obtained similarly to compound 28 from comꢀ
pound 23. The yield was 87%. White crystals, m.p. 90—92 C
(
10—15 mL) and the solvent was evaporated in vacuo. The resiꢀ
due was purified by column chromatography on SiO2 (eluent
hexane—EtOAc) to obtain compound 25 (0.13 g, 57%) as white
(EtOH), []D20 –22.1 (c 0.24, CHCl ). Found (%): C, 68.35;
3

Lupane triphenylphosphonium compounds
Russ.Chem.Bull., Int.Ed., Vol. 62, No. 1, January, 2013
197
H, 9.56; S, 4.97. C H O S. Calculated (%): C, 68.27; H, 9.60;
H(23)—H(27)); 1.11—2.28 (m, 22 H, CH, CH in pentacyclic
3
7
62
7
2
S, 4.93. IR, /cm–1: 1357 (S=O), 1729 (C=O). H NMR, : 0.75,
1
skeleton, 1 H, H(20), 2 H, H(1´), 2 H, H(2´)); 1.67 (m, 1 H,
b
0
.85 (both d, 3 H each, H(29), H(30), J = 7 Hz); 0.81, 0.86,
H
ax(1)); 2.12 (s, 3 H, Me (OAc)); 3.36 (t, 2 H, H(3´), J = 7 Hz);
0
.88, 0.91, 0.99 (all s, 3 H each, H(23)—H(27)); 1.10—2.24
4.45 (d, 1 H, H(3), J = 11 Hz); 5.12 (m, 2 H, OCH Ph);
2
+
(
m, 24 H, CH, CH in pentacyclic skeleton, 1 H, H(20), 2 H,
7.28—7.35 (m, 5 H, Ph). MS, m/z: 735.4 [M + Na] . C H O Br.
2
42 63
4
+
H(1´), 2 H, H(2´)); 2.10 (s, 3 H, Me (OAc)); 3.00 (s, 3 H, OMs);
.65 (s, 3 H, OMe); 4.11—4.20 (m, 2 H, H(3´)); 4.75 (br.s, 1 H,
Calculated, m/z: 735.4 [M + Na] .
3
3ꢀAcetoxyꢀ2ꢀ(3ꢀbromopropyl)ꢀ20,29ꢀdihydrobetulinic acid
(34) was obtained from compound 33 according to a typical
+
+
H(3)). MS, m/z: 673.5 [M + Na] , 689.5 [M + K] . C H O S.
Calculated, m/z: 673.4 [M + Na] , 689.4 [M + K] .
Benzyl 3ꢀacetoxyꢀ2ꢀ(3ꢀmesyloxypropyl)ꢀ20,29ꢀdihydroꢀ
betulinate (30) was obtained similarly to compound 28 from comꢀ
pound 24. The yield was 90%. White crystals, m.p. 78—80 C
3
7
62
7
+
+
19
procedure. The yield was 97%. White crystals, m.p. 184—186 C
(EtOH), []D20 –34.4 (c 0.42, CHCl ). Found (%): C, 67.70;
3
H, 9.19; Br, 12.78. C H O Br. Calculated (%): C, 67.61; H, 9.24;
3
5
57 4
–
1
1
Br, 12.85. IR, /cm : 1731 (C=O), 3366 (OH). H NMR,
: 0.69 (t, 1 H, H eq(1), J = 12 Hz); 0.77, 0.87 (both d, 3 H each,
2
0
a
(
EtOH), []D –22.3 (c 0.26, CHCl ). Found (%): C, 70.81;
3
H, 9.00; S, 4.45. C H O S. Calculated (%): C, 71.04; H, 9.15;
H(29), H(30), J = 7 Hz); 0.78, 0.83, 0.89, 0.95, 0.97 (all s,
4
1
3
66
7
–
1
S, 4.41. IR, /cm : 1358 (S=O), 1730 (C=O). H NMR, : 0.67
3 H each, H(23)—H(27)); 1.10—2.29 (m, 22 H, CH, CH in
2
a
(
t, 1 H, H eq(1), J = 12 Hz); 0.76, 0.86 (both d, 3 H each, H(29),
pentacyclic skeleton, 1 H, H(20), 2 H, H(1´), 2 H, H(2´)); 1.68
(m, 1 H, Hbax(1)); 2.12 (s, 3 H, Me (OAc)); 3.36 (t, 2 H, H(3´),
J = 7 Hz); 4.47 (d, 1 H, H(3), J = 11 Hz). MS, m/z: 643.4
H(30), J = 7 Hz); 0.77, 0.81, 0.83, 0.87, 0.94 (all s, 3 H each,
H(23)—H(27)); 1.08—1.91 (m, 22 H, CH, CH in pentacyclic
2
+
+
skeleton, 1 H, H(20), 2 H, H(1´), 2 H, H(2´)); 1.68 (m, 1 H,
[M + Na] . C H O Br. Calculated, m/z: 643.3 [M + Na] .
35 57 4
b
H
ax(1)); 2.11 (s, 3 H, Me (OAc)); 3.01 (s, 3 H, OMs); 4.18
Methyl 3ꢀacetoxyꢀ2ꢀ(3ꢀtriphenylphosphoniopropyl)ꢀ20,29ꢀ
dihydrobetulinate bromide (3). A mixture of bromide 31 (0.24 mmol),
(
(
t, 2 H, H(3´), J = 6 Hz); 4.45 (d, 1 H, H(3), J = 11 Hz); 5.12
m, 2 H, OCH Ph); 7.28—7.37 (m, 5 H, Ph).
toluene (10 mL), and Ph P (0.72 mmol) was refluxed for 32 h
2
3
Methyl 3ꢀacetoxyꢀ2ꢀ(3ꢀbromopropyl)ꢀ20,29ꢀdihydrobetulꢀ
inate (31). Lithium bromide (0.07 g, 0.8 mmol) was added to
a solution of mesylate 28 (0.42 mmol) in anhydrous acetone
12 mL). The mixture was refluxed for 3 h (TLC monitoring)
and cooled, a residue formed was filtered and washed with acetꢀ
one (2 mL). A combined filtrate was concentrated in vacuo. The
(TLC monitoring). The solution was cooled and the solvent was
evaporated in vacuo. A solid product obtained was washed with
hot hexane (2×7 mL), dissolved in EtOAc (2 mL) and diluted
with hexane (8 mL). The precipitate was filtered off to obtained
compound 3 (0.20 g, 94%) as white crystals, m.p. 165—167 C
(
2
0
–1
(EtOH), []
–22.3 (c 0.95, CHCl ). IR, /cm : 1727 (C=O).
D
3
1
a
residue was purified by column chromatography on SiO (eluent
H NMR, : 0.46 (t, 1 H, H eq(1), J = 12 Hz); 0.74, 0.76 (both d,
3 H each, H(29), H(30), J = 7 Hz); 0.75, 0.77, 0.79, 0.87, 0.89
(all s, 3 H each, H(23)—H(27)); 1.17—2.22 (m, 22 H, CH, CH2
in pentacyclic skeleton, 1 H, H(20), 2 H, H(1´), 2 H, H(2´));
1.65 (m, 1 H, Hbax(1)); 2.06 (s, 3 H, Me(OAc)); 3.63 (s, 3 H,
OMe); 3.82 (m, 2 H, H(3´)); 4.30 (d, 1 H, H(3), J = 11 Hz);
2
hexane—EtOAc) to obtain compound 31 (0.21 g, 79%) as white
crystals, m.p. 92—94 C (EtOH), []D2 +19.2 (c 1.50, CHCl ).
0
3
Found (%): C, 68.21; H, 9.35; Br, 12.72. C H O Br. Calculatꢀ
3
6
59
4
–
1
ed (%): C, 68.01; H, 9.35; Br, 12.57. IR, /cm : 1732 (C=O).
1
a
H NMR, : 0.66 (t, 1 H, H eq(1), J = 12 Hz); 0.76, 0.86 (both d,
H each, H(29), H(30), J = 7 Hz); 0.81, 0.83, 0.88, 0.92, 0.95
all s, 3 H each, H(23)—H(27)); 1.13—2.25 (m, 22 H, CH, CH2
in pentacyclic skeleton, 1 H, H(20), 2 H, H(1´), 2 H, H(2´));
31
3
(
7.70—7.87 (m, 15 H, Ph) P NMR, : 24.00. MS, m/z: 817.4
+
+
[M – Br] . C H O BrP. Calculated, m/z: 817.5 [M – Br] .
54 74 4
Methyl 3ꢀacetoxyꢀ2ꢀ(3ꢀtriphenylphosphoniopropyl)ꢀ20,29ꢀ
dihydrobetulinate bromide (4) was obtained similarly to compound 3
from bromide 32. The yield was 89%. White crystals, m.p.
b
1
.66 (m, 1 H, H ax(1)); 2.11 (s, 3 H, Me (OAc)); 3.36 (t, 2 H,
H(3´), J = 7 Hz); 3.66 (s, 3 H, OMe); 4.46 (d, 1 H, H(3),
+
20
–1
J = 11 Hz). MS, m/z: 634.1 [M] . C H O Br. Calculated,
m/z: 634.3 [M] .
170—172 C (EtOH), []
+0.2 (c 0.42, CHCl ). IR, /cm :
3
6
59
4
D
3
+
1
1726 (C=O). H NMR, : 0.75, 0.86 (both d, 3 H each, H(29),
H(30), J = 7 Hz); 0.77, 0.78, 0.80, 0.88, 0.97 (all s, 3 H each,
Methyl 3ꢀacetoxyꢀ2ꢀ(3ꢀbromopropyl)ꢀ20,29ꢀdihydrobetulꢀ
inate (32) was obtained similarly to compound 31 from comꢀ
H(23)—H(27)); 1.02—2.30 (m, 24 H, CH, CH in pentacyclic
2
pound 29. The yield was 78%. White crystals, m.p. 84—86 C
skeleton, 1 H, H(20), 2 H, H(1´), 2 H, H(2´)); 2.10 (s, 3 H, Me
(OAc)); 3.46, 4.01 (both m, 2 H, H(3´)); 3.64 (s, 3 H, OMe);
EtOH), []D2 +20.9 (c 0.46, CHCl ). Found (%): C, 68.10;
0
(
3
31
H, 9.29; Br, 12.61. C H O Br. Calculated (%): C, 68.01; H, 9.35;
4.54 (br.s, 1 H, H(3)); 7.64—7.81 (m, 15 H, Ph). P NMR,
3
6
59
4
–
1
1
+
Br, 12.57. IR, /cm : 1731 (C=O). H NMR, : 0.76, 0.87
both d, 3 H each, H(29), H(30), J = 7 Hz); 0.83, 0.88, 0.89,
.93, 1.01 (all s, 3 H each, H(23)—H(27)); 1.11—2.28 (m, 24 H,
: 23.77. MS, m/z: 817.5 [M – Br] . C H O BrP. Calculated,
m/z: 817.5 [M – Br] .
5
4
74
4
+
(
0
Methyl 3ꢀhydroxyꢀ2ꢀ(3ꢀtriphenylphosphoniopropyl)ꢀ20,29ꢀ
dihydrobetulinate iodide (5) was obtained similarly to compound 3
CH, CH in pentacyclic skeleton, 1 H, H(20), 2 H, H(1´), 2 H,
2
H(2´)); 2.10 (s, 3 H, Me (OAc)); 3.28—3.40 (m, 2 H, H(3´));
by the reaction of iodide 25 with Ph P for 16 h. The yield was
3
+
20
3
.65 (s, 3 H, OMe); 4.76 (br.s, 1 H, H(3)). MS, m/z: 634.2 [M] .
21%. White crystals, m.p. 124—126 C (EtOH), []
–21
D
+
–1
1
C H O Br. Calculated, m/z: 634.3 [M] .
(c 0.22, CHCl ). IR, /cm : 1724 (C=O). H NMR, : 0.48
3
6
59
4
3
a
Benzyl 3ꢀacetoxyꢀ2ꢀ(3ꢀbromopropyl)ꢀ20,29ꢀdihydrobetulꢀ
(t, 1 H, H eq(1), J = 12 Hz); 0.75, 0.87 (both d, 3 H each, H(29),
inate (33) was obtained similarly to compound 31 from comꢀ
H(30), J = 7 Hz); 0.74, 0.76, 0.79, 0.85, 0.88 (all s, 3 H each,
pound 30. The yield was 81%. White crystals, m.p. 86—88 C
H(23)—H(27)); 1.13—2.30 (m, 22 H, CH, CH in pentacyclic
2
2
0
(
EtOH), []D –30.5 (c 0.21, CHCl ). Found (%): C, 70.91;
skeleton, 1 H, H(20), 2 H, H(1´), 2 H, H(2´)); 1.65 (m, 1 H,
3
b
H, 8.88; Br, 11.25. C H O Br. Calculated (%): C, 70.86;
H
(1)); 2.89 (d, 1 H, H(3), J = 10 Hz); 3.60 (m, 2 H, H(3´));
4
2
63
4
ax
–
1
1
31
H, 8.92; Br, 11.22. IR, /cm : 1732 (C=O). H NMR, : 0.67
t, 1 H, H eq(1), J = 12 Hz); 0.75, 0.87 (both d, 3 H each, H(29),
3.64 (s, 3 H, OMe); 7.60—7.90 (m, 15 H, Ph). P NMR,
: 24.08. MS, m/z: 775.4 [M – I] . C H O IP. Calculated,
m/z: 775.5 [M – I] .
a
+
(
52
72
3
+
H(30), J = 7 Hz); 0.76, 0.82, 0.83, 0.86, 0.94 (all s, 3 H each,

1
98
Russ.Chem.Bull., Int.Ed., Vol. 62, No. 1, January, 2013
Spivak et al.
Methyl 3ꢀhydroxyꢀ2ꢀ(3ꢀtriphenylphosphoniopropyl)ꢀ20,29ꢀ
dihydrobetulinate iodide (6) was obtained similarly to compound 5
T. J. Hieken, T. K. Das Gupta, J. M. Pezzuto, Nat. Med.,
1995, 1, 1046.
from iodide 26. The yield was 23%. White crystals, m.p.
5. R. A. Cichewicz, S. A. Kouzi, Med. Res. Rev., 2004, 24, 90.
6. R. Mukherjee, V. Kumar, S. K. Srivastava, S. K. Agarwal,
A. C. Burman, AntiꢀCancer Agents Med. Chem., 2006, 6, 271.
7. S. Fulda, Int. J. Mol. Sci., 2008, 9, 1096.
2
0
–1
1
1
42—144 C (EtOH), []
725 (C=O). H NMR, : 0.75, 0.87 (both d, 3 H each, H(29),
–1 (c 0.20, CHCl ). IR, /cm :
3
D
1
H(30), J = 7 Hz); 0.76, 0.79, 0.85, 0.88, 0.93 (all s, 3 H each,
H(23)—H(27)); 1.13—1.80 (m, 24 H, CH, CH in pentacyclic
8. J. Neuzil, L.ꢀF. Dong, L. Ramanathapuram, T. Hahn,
M. Chladova, X.ꢀF. Wang, R. Zobalova, L. Prochazka, M. Gold,
R. Freeman, J. Turanek, E. T. Akporiaye, J. C. Dyason, S. J.
Ralphf, Mol. Aspects Med., 2007, 28, 607.
9. S. Fulda, G. Kroemer, Drug Discovery Today, 2009, 14, 885.
10. F. B. Mullauer, J. H. Kessler, J. P. Medema, Anticancer
Drugs, 2010, 21, 215.
2
skeleton, 1 H, H(20), 2 H, H(1´), 2 H, H(2´)); 3.56 (m, 2 H,
H(3´)); 3.64 (s, 3 H, OMe); 5.15 (br.s, 1 H, H(3)); 7.67—7.89
(
m, 15 H, Ph). 3 P NMR, : 24.34. MS, m/z: 775.3 [M – I] .
1
+
+
C H O IP. Calculated, m/z: 775.5 [M – I] .
5
2
72
3
3
ꢀAcetoxyꢀ2ꢀ(3ꢀtriphenylphosphoniopropyl)ꢀ20,29ꢀdiꢀ
hydrobetulinic acid bromide (7) was obtained similarly to compound
3
2
1
0
0
from bromo acid 34. The yield was 81%. White crystals, m.p.
11. F. Wang, M. A. Ogasawara, P. Huang, Mol. Aspects Med.,
2010, 31, 75.
12. L. Biasutto, L.ꢀF. Dong, M. Zoratti, J. Neuzil, Mitochonꢀ
drion, 2010, 10, 670.
13. B. Bachowska, J. KazmierczakꢀBaranska, M. Cieslak,
B. Nawrot, D. Szczesna, J. Skalik, P. Balczewski, Chemistryꢀ
open, 2012, 1, 33.
2
0
–1
06—208 C (EtOH), []D –25.4 (c 0.24, CHCl ). IR, /cm
:
3
1
a
726 (C=O). H NMR, : 0.47 (t, 1 H, H eq(1), J = 12 Hz); 0.74,
.86 (both d, 3 H each, H(29), H(30)); 0.71, 0.73, 0.80, 0.85,
.88 (all s, 3 H each, H(23)—H(27)); 1.12—2.21 (m, 22 H, CH,
CH2 in pentacyclic skeleton, 1 H, H(20), 2 H, H(1´), 2 H,
H(2´)); 1.68 (m, 1 H, H ax(1)); 2.03 (s, 3 H, Me (OAc));
b
3
.62—3.75 (m, 2 H, H(3´)); 4.29 (d, 1 H, H(3), J = 11 Hz); 7.28
14. M. Millard, D. Pathania, Y. Shabaik, L. Taheri, J. Deng,
N. Neamati, PLoS ONE, 2010, 5, e13131.
15. A. Manetta, G. Gamboa, A. Nasseri, Y. D. Podnos, D. Emꢀ
ma, G. Dorion, L. Rawlings, P. M. Carpenter, A. Bustaꢀ
mante, J. Patel, D. Rideout, Gynecologic Oncology, 1996,
60, 203.
16. S. J. Ralph, J. Neuzil, Patent Aplication Publication № US
2011/ 0105437 A1; http://www.faqs.org/patents/app/
20110105437.
17. DSHL. Kim, Z. Chen, V. T. Ngugen, J. M. Pezzuto, S. Qui,
Z.ꢀZ. Lu, Synt. Commun., 1997, 27, 1607.
3
1
(
br.s, 1 H, COOH); 7.47—7.98 (m, 15 H, Ph). P NMR,
+

: 24.15. MS, m/z: 803.4 [M – Br] . C H O BrP. Calculated,
5
3
72
4
+
m/z: 803.5 [M – Br] .
Benzyl 3ꢀacetoxyꢀ2ꢀ(3ꢀtriphenylphosphoniopropyl)ꢀ20,29ꢀ
dihydrobetulinate iodide (8) was obtained similarly to compound 5
from iodide 27. The yield was 78%. White crystals, m.p.
1
1
0
0
2
0
–1
48—150 C (EtOH), []D –17.9 (c 0.24, CHCl ). IR, /cm :
3
1
a
728 (C=O). H NMR, : 0.47 (t, 1 H, H eq(1), J = 12 Hz); 0.70,
.86 (both d, 3 H each, H(29), H(30)); 0.71, 0.74, 0.77, 0.86,
.88 (all s, 3 H each, H(23)—H(27)); 1.04—2.27 (m, 22 H, CH,
CH2 in pentacyclic skeleton, 1 H, H(20), 2 H, H(1´), 2 H,
H(2´)); 1.65 (m, 1 H, H ax(1)); 2.07 (s, 3 H, Me (OAc));
18. M. Urban, J. Sarek, J. Klinot, G. Kornikova, M. Hajduch,
J. Nat. Prod., 2004, 67, 1100.
b
3
5
.53—3.66 (m, 2 H, H(3´)); 4.31 (d, 1 H, H(3), J = 11 Hz);
19. C. Genet, A. Strehle, C. Schmidt, G. Boudjelal, A. Lobꢀ
stein, K. Schoonjans, M. Souchet, J. Auwerx, R. Saladin,
A. Wagner, J. Med. Chem., 2010, 53, 178.
20. A. Yu. Spivak, E. R. Shakurova, D. A. Nedopekina, R. R.
Khalitova, L. M. Khalilov, V. N. Odinokov, Yu. P. Bel´skii,
A. N. Ivanova, N. V. Bel´skaya, M. G. Danilets, A. A.
Ligacheva, Russ. Chem. Bull. (Int. Ed.), 2011, 60, 694 [Izv.
Akad. Nauk, Ser. Khim., 2011, 680].
.05—5.13 (m, 2 H, OCH Ph); 7.29—7.85 (m, 20 H, Ph).
2
3
1
+
P NMR, : 24.11. MS, m/z: 893.5 [M – I] . C H O IP.
6
0
78
4
+
Calculated, m/z: 893.6 [M – I] .
This work was financially supported by the Russian
Foundation for Basic Research (Project Nos 10ꢀ03ꢀ00105
and 12ꢀ03ꢀ97005ꢀa) and the Russian Academy of Sciencꢀ
es (Program of the Division of Chemistry and Materials
Science of RAS "Medicinal Chemistry: Molecular Deꢀ
sign of Physiologically Active Compounds and Mediciꢀ
nal Drugs").
2
1. O. B. Flekhter, L. R. Nigmatullina, L. A. Baltina, L. T.
Karachurina, F. Z. Galin, F. S. Zarudii, G. A. Tolstikov,
E. I. Boreko, N. I. Pavlova, S. N. Nikolaeva, O. V. Saviꢀ
nova, Khim. Farm. Zh., 2002, 36, 26 [Pharm. Chem. J.
(
Engl. Transl.), 2002, 36].
2
2
2. J. L. Luche, J. Am. Chem. Soc., 1978, 100, 2226.
3. E. St´astna, I. Cerny, V. Pouzar, H. Chodounska, Steroids,
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3
4
Received November 1, 2012;
in revised form December 24, 2012