Conjugates of betulin derivatives with AZT as potent anti-HIV agents

Article abstract of DOI:10.1016/j.bmc.2010.06.092

Fourteen novel conjugates of 3,28-di-O-acylbetulins with AZT were prepared as anti-HIV agents, based on our previously reported potent anti-HIV triterpene leads, including 3-O-acyl and 3,28-di-O-acylbetulins. Nine of the conjugates (49-53, 55, 56, 59, and

Full text of DOI:10.1016/j.bmc.2010.06.092

Bioorganic & Medicinal Chemistry 18 (2010) 6451–6469
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Conjugates of betulin derivatives with AZT as potent anti-HIV agents
b
c
c
Juan Xiong a, Yoshiki Kashiwada a, , Chin-Ho Chen , Keduo Qian , Susan L. Morris-Natschke ,
*
Kuo-Hsiung Lee c, , Yoshihisa Takaishi
a
*
a Graduate School of Pharmaceutical Sciences, University of Tokushima, Tokushima 770-8505, Japan
b Medical Center, Duke University, Durham, NC 27710, USA
c Natural Products Research Laboratories, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC 27599-7568, USA
a r t i c l e i n f o
a b s t r a c t
Article history:
Fourteen novel conjugates of 3,28-di-O-acylbetulins with AZT were prepared as anti-HIV agents, based on
our previously reported potent anti-HIV triterpene leads, including 3-O-acyl and 3,28-di-O-acylbetulins.
Nine of the conjugates (49–53, 55, 56, 59, and 60) exhibited potent anti-HIV activity at the submicrom-
Received 11 June 2010
Revised 28 June 2010
Accepted 29 June 2010
Available online 7 July 2010
olar level, with EC50 values ranging from 0.040 to 0.098 lM in HIV-1NL4-3 infected MT-4 cells. These com-
pounds were equipotent or more potent than 3-O-(30,30-dimethylsuccinyl)betulinic acid (2), which is
currently in Phase IIb anti-AIDS clinical trial.
Keywords:
HIV-1
Ó 2010 Elsevier Ltd. All rights reserved.
Betulin
AZT
Conjugate
1. Introduction
more efficacious and less prone to resistance than monothera-
pies.16,17 Although it is more challenging to create multi-target
Betulinic acid (1), a lupane-type triterpene, is widely distributed
throughout the plant kingdom. Various biological activities1–5 have
been reported for 1, including anti-HIV, anti-cancer, and anti-
inflammatory properties. Our previous modification study on 1
led to the discovery of 3-O-(30,30-dimethylsuccinyl)betulinic acid
(2, bevirimat, now also known as MPC-4326). Compound 2 exhibits
potent anti-HIV activity and is currently in Phase IIb clinical trials
in the US.6–9 It is a first-in-class drug candidate as a viral matura-
tion inhibitor, which disrupts the processing of capsid precursor
p25 (CA-SP1) to mature capsid protein p24 (CA), resulting in gen-
eration of non-infectious HIV-1 virions.10
therapeutics by building multi-target actions into one single chem-
ical entity rather than mixing monotherapies,16,18 the multi-tar-
geted single agent could greatly simplify treatment regimens and
reduce the risk of drug–drug interactions.16 Herein, we report
our preliminary research on conjugation of 30-azido-30-deoxythy-
midine (AZT),19 the first clinically approved nucleoside reverse
transcriptase inhibitor (NRTI), with betulin derivatives. The hybrid
conjugates were formed through a linker with ester bonds, which
are easy to hydrolyze and subsequently release the parent com-
pounds, betulin derivative and AZT, to exert their functions.
In subsequent research, various derivatives of betulin (3a) and
dihydrobetulin (3b) were investigated based on the structural sim-
ilarity of 1 and 3a, 3b.11–14 Several analogs, including 3-O-glut-
aryldihydrobetulin (4),14 3,28-di-O-(30,30-dimethylglutaryl)betulin
(5), and 3-O-(30,30-dimethylsuccinyl)-28-O-(200,200-dimethylsucci-
nyl)betulin (6)12,13 exhibited comparable or greater anti-HIV activ-
ity compared with the structurally related 215 (Fig. 1).
In a continuing study of potent anti-HIV agents based on the
betulin scaffold, we wanted to explore the conjugation of our un-
ique HIV-1 maturation inhibitors with other classes of anti-HIV
agents, because the strategy of multi-target therapeutics could be
2. Design
Initially, we considered where to link AZT to the triterpene skel-
eton. 3,28-Di-O-acylbetulin derivatives contain two free carboxylic
acid groups, which appear to be ideal fragments for constructing
conjugates. Previous studies suggested that the free terminal car-
boxylic acid in the 3-O-acyl group is essential for anti-maturation
activity; thus, the other –COOH group in the 28-O-acyl moiety
was considered to be the appropriate conjugating site (Fig. 2).
Secondly, we considered the identity of the 3-O-acyl group (R1).
Our prior modification studies on 1, 3a, and 3b showed that the
esterification of the 3-OH with 30,30-dimethylglutaryl or 30,30-dim-
ethylsuccinyl fragments produced significantly active derivatives
(e.g., 5 and 6).6,12,14,20–22 However, the 3a-derivatives bearing a
3-O-glutaryl group exhibited only weak or moderate anti-HIV
activity.14 In contrast, introduction of a glutaryl group onto the
* Corresponding authors. Tel./fax: +81 88 633 7276 (Y.K.); tel.: +1 919 962 0066;
fax: +1 919 966 3893 (K.-H.L.).
E-mail addresses: kasiwada@ph.tokukshima-u.ac.jp (Y. Kashiwada), khlee@
unc.edu (K.-H. Lee).
0968-0896/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2010.06.092

6452
J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
H
H
H
COOH
H
COOH
O
H
H
HO
HO
O
H
H
O
Betulinic Acid (1)
Bevirimat (2)
29
20
30
H
H
OH
OH
H
H
28
O
O
H
H
3
HO
HO
O
H
H
4
Betulin (3a): C20=C29, double bond
Dihydrobetulin (3b): C20-C29, single bond
O
OH
H
R1=
R2=
R2=
5
6
OR2
HO
O
O
O
H
O
H
HO
R1=
OH
R1
O
H
O
O
Figure 1. Betulinic acid, betulin and their derivatives which demonstrated potent anti-HIV activities.
for constructing the triterpene-AZT conjugates. Succinyl, glutaryl,
and 3,3-dimethylsuccinyl esters were also investigated as the 28-
O-acyl moiety in the conjugate compounds.
The preparation of 14 new conjugates of 3,28-di-O-acylbetulins
with AZT and evaluation of anti-HIV activity are described in this
paper. Table 1 lists the structural identities of the 3- and 28-O-acyl
side chains, and the degree of saturation of the C20–C29 bond for all
intermediates (21–48) and final conjugates (49–62).
O
N
HN
O
H
R2
O
OH
H
HO
O
O
O
H
R1O
H
N3
AZT
3,28-di-O-acylbetulins
3. Chemistry
O
N
The synthetic route to conjugated products (49–62) is outlined
in Schemes 1 and 2. The key intermediate 3-O-acylbetulins (11–13)
were successfully prepared by the methods described previously14
as shown in Scheme 1. Protection of the 28-OH group with triphen-
ylmethyl (or trityl) chloride yielded betulin 28-O-trityl ether (7),
which was further treated with an appropriate dicarboxylic acid
or anhydride in anhydrous pyridine in the presence of 4-dimethyl-
aminopyridine (DMAP) to furnish the corresponding 3-O-acyl-
28-O-tritylbetulins (8–10). As was found in previous studies, the
reaction of 7 with 2,2-dimethylsuccinic acid gave a mixture
of 3-O-20,20-dimethylsuccinyl- and -30,30-dimethylsuccinylbetulin
derivatives, in which the latter isomer (8) was the major product.
The isomers were separated by silica gel chromatography, and
their structures were assigned by 2D NMR analysis (HMBC). For
8 and 9, the protecting trityl group was subsequently cleaved with
pyridium p-toluenesulfonate (PPTS) in CH2Cl2–EtOH to yield 3-O-
acylbetulins (11 and 12, respectively). An ethyl ester side-product
was obtained predominately by removing the 28-O-trityl group
from 3-O-glutaryl-28-O-tritylbetulin (10) with PPTS; however,
the desired product (13) was obtained exclusively by refluxing
10 with Lewis acid FeCl3 in 95% THF.
HN
H
O
R2
O
O
H
O
O
O
H
R1O
N3
H
Figure 2. General structure of hybrid-type anti-HIV agents.
3-OH of 3b led to a dramatic increase of anti-HIV potency as seen
in compound 4. Therefore, in this study, triterpenes with the lup-
20(29)-ene scaffold of 3a were acylated with 3,3-dimethylglutaryl
(54) and 3,3-dimethylsuccinyl (49–53) moieties, while those with
the lupane scaffold of 3b were also acylated with a glutaryl group
(58–61), as well as the two preceding moieties (55–57 and 62).
Thirdly, we considered the identity of the 28-O-acyl group (R2).
Although previous biological data for betulin derivatives suggested
that the group at this position might be not essential for the anti-
HIV activity of this compound type, it could still have some influ-
ence on potency.12 In addition, here the 28-O-acyl group is also
being used as a linker in the conjugates. Therefore, the identity
of the linking ester group might play a vital part in the anti-HIV
activity of hybrid conjugates. Among the 3,28-di-O-acylbetulin
derivatives previously studied, compounds 5 and 6 exhibited the
greatest potency, and thus the 3,3-dimethylglutaryl and 2,2-dim-
ethylsuccinyl groups found in these two compounds were used
Hydrogenation of 3-O-acylbetulins (11–13) with H2/Pd–C in
EtOAc–EtOH afforded the 3-O-acyldihydrobetulins (14, 15, and 4,
respectively) almost quantitatively. Before a second acyl moi-
ety was introduced at the C-28 position, the free terminal carbox-
ylic acid in the 3-O-acyl group was first protected as the

J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
6453
diphenyldiazomethyl ester. Treatment of 3-O-acylbetulins (11, 12)
and -dihydrobetulins (14, 15, 4) with freshly prepared diphenyl-
diazomethane in MeOH produced the protected derivatives (16–
20) in quantitative yield.
conjugates (35–48). However, the corresponding anhydride was
also obtained in each case, resulting in only a moderate yield of
the desired conjugate. Finally, the terminal protecting group on
the 3-O-acyl moiety was removed with 80% acetic acid to afford
the target conjugates (49–62).
The 3,28-di-O-acylbetulins (21–34) were readily prepared by
heating the protected 3-O-acylbetulin derivatives with an appro-
priate dicarboxylic acid or anhydride overnight in the presence of
DMAP and dry pyridine (Scheme 2). The two isomeric products ob-
tained when a dimethylsuccinyl group was introduced at C-28 of
16, 18, and 20 were successfully separated by preparative HPLC,
yielding corresponding pure derivatives. The orientation of the
dimethylsuccinyl group attached to the C-28 hydroxy group of
22/23, 27/28, and 30/31 was established by 2D NMR analyses
(HSQC and HMBC). The 28-O-(300,300-dimethylsuccinyl)betulin
derivative was the major product in each case. The electron-donat-
ing effect of the dimethyl groups on the adjacent carboxyl group
would cause decreased electrophilicity, resulting in different yields
of the two isomers in acylation reactions either at C-3 or C-28.
Conjugation of the 3,28-di-O-acylbetulin (21–26) and -dihydro-
betulin derivatives (27–34) with AZT in the presence of dicyclohex-
ylcarbodiimide (DCC) and DMAP in CH2Cl2 gave the corresponding
4. Results and discussion
The anti-HIV-1 replication activities of the newly synthesized
conjugates (49–62) were evaluated in HIV-1NL4-3 infected MT-4
cells in parallel with AZT. The bioassay results are summarized in
Table 2.
In the MT-4 screening system, nine conjugates (49–53, 55, 56,
59, and 60) exhibited potent anti-HIV activity at a submicromolar
level (EC50 values ranging from 0.040 to 0.098 lM), and thus, were
equipotent or more potent than 2. Compounds 51 and 49, which
have a 3-O-(30,30-dimethylsuccinyl) moiety together with a 28-O-
(200,200-dimethylsuccinyl) or 28-O-succinyl group, respectively,
linked to AZT, exhibited the highest potency, with EC50 values of
0.040 and 0.045 lM, respectively. Interestingly, all three conju-
gates (51, 56, and 59) with a 2,2-dimethylsuccinyl linking group
H
H
H
OH
O
O
a
H
b
H
H
C(C6H5)3
C(C6H5)3
O
H
H
H
HO
HO
O
R
H
H
H
HOOC
R=
R=
R=
8
9
Betulin (3a)
7
HOOC
HOOC
10
H
H
H
H
H
OH
OH
OH
e
c
H
d
O
O
O
H
H
H
(C6H5)2HC
O
R
O
O
R
R
H
H
H
O
R=
R=
R=
14
R=
11
12
HOOC
HOOC
HOOC
HOOC
R=
R=
R=
18
O
O
15
4
R=
R=
19
20
O
13
HOOC
HOOC
O
O
e
H
OH
H
O
H
(C6H5)2HC
O
R
H
O
16
R=
R=
O
O
17
O
Scheme 1. Reagents and conditions: (a) trityl chloride, DAMP, DMF, reflux; (b) dicarboxylic acid (or anhydride), DMAP, pyridine, reflux; (c) PPTS, CH2Cl2–EtOH or FeCl3, 95%
THF, reflux; (d) H2, 10% Pd/C, EtOAc–EtOH, rt; (e) diphenyldiazomethane, MeOH, rt.

6454
J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
H
H
dicarboxylic acid
OR2
OH (or anhydride), DMAP
DCC, DMAP, AZT
CH2Cl2
H
H
pyridine
H
H
R1
R1
(C6H5)2HC
(C6H5)2HC
O
O
H
H
21-34
16-20
O
N
O
HN
HN
H
H
H
O
O
N
O
O
80% HOAc
H
R2
R2
O
O
H
H
H
R1
R1
(C6H5)2HC
O
O
H
N3
N3
49-62
35-48
Scheme 2. General synthetic route to conjugates of betulin derivatives with AZT.
Table 1
Ester side chains of 21–62.
Table 2
Anti-HIV-1 data of 49–62 against HIV-1NL4-3 infected MT-4 cellsa
b
c
Compound
C20–C29
R1
R2
Compound
EC50
(l
M)
IC50 (lM)
TId
158
71
135
145
129
43
103
138
31
28
110
97
21, 35, 49
22, 36, 50
23, 37, 51
24, 38, 52
25, 39, 53
26, 40, 54
C@C
Bb
B
B
B
B
Aa
B
49
50
51
52
53
54
55
56
57
58
59
60
61
62
2
0.045
0.098
0.040
0.060
0.063
0.18
0.087
0.056
0.29
7.1
7.0
5.4
8.7
8.1
7.7
9.0
7.7
9.0
7.2
8.0
9.0
8.5
9.8
c
B2
Cd
De
D
D
27, 41, 55
28, 42, 56
29, 43, 57
30, 44, 58
31, 45, 59
32, 46, 60
33, 47, 61
34, 48, 62
CAC
B
B
B
C
C
C
C
D
B
B2
D
B
B2
C
0.26
0.073
0.093
1.21
7
27
D
D
0.36
0.096e
0.027
>5e
>37.4
>52e
>1385
a
AZT
A, succinyl
a
b
c
Data presented are averages of at least two separate experiments.
Concentration that inhibits HIV-1NL4-3 replication by 50%.
Concentration that inhibits mock-infected MT-4 cell growth by 50%.
d
e
TI = IC50/EC50
.
b
B, 3,3-dimethylsuccinyl
Data are taken from Ref. 23.
linking unit at C-28 showed decreased activity in HIV-1 infected
MT-4 cells relative to those with a 2,2-dimethylsuccinyl group
(51, 56, and 59). These results confirmed our previous finding that
a 2,2-dimethylsuccinyl is superior to a 3,3-dimethylsuccinyl side
chain at the C-28 position of anti-HIV triterpene analogs. For exam-
ple, compound 6 was reported to be 230-fold more potent against
HIV replication in H9 lymphocytes assay than its isomer with a 3,3-
dimethylsuccinyl side chain at C-28.13
c
B2, 2,2-dimethylsuccinyl
d
e
C, glutaryl
Among conjugates containing glutaryl or dimethyglutaryl es-
D, 3,3-dimethylglutaryl
ters, 59 and 60 [3-O-glutaryl, 28-O-(200,200-dimethylsuccinyl), EC50
0.073 lM and 3,28-di-O-glutaryl, EC50 0.093 lM] were equipotent
with 2. However, 54 and 62 [3,28-di-O-(30,30-dimethylglutaryl)]
showed only moderate activity, and 61 [3-O-glutaryl, 28-O-
(300,300-glutaryl)] was the weakest compound.
at C-28 generally showed higher potency than their corresponding
congeners with the same 3-O-acyl group, but a different 28-O-acyl
group (51 compared with 50, 52, and 53; 56 compared with 55 and
57; 59 compared with 58, 60, and 61). Of particular note, the con-
jugates (50, 55, and 58) with a 3,3-dimethylsuccinyl moiety as the
Comparison of the anti-HIV potency of dihydrobetulin conju-
gates 55–57 with that of betulin conjugates 50, 51, and 53 sug-
gested that hydrogenation of the isopropylidene group at C-19
did not help to increase the anti-HIV potency. Finally, comparison
of data for 49/63 and 52/64 in Table 3 showed that hybrid conju-

J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
6455
Table 3
(3H, m, H-22, H-16 and H-19), 2.92 (1H, d, J = 8.8 Hz, H-28),
3.15–3.19 (2H, m, H-28 and H-3), 4.53, 4.59 (each 1H, br s, H2-
29), 7.23 (3H, t, J = 7.2 Hz, trityl H-40), 7.31 (6H, t, J = 7.6, 7.2 Hz, tri-
tyl H-30, 50), 7.49 (6H, d, J = 7.6 Hz, trityl H-20, 60); 13C NMR (CDCl3,
100 MHz) d 14.7 (C-27), 15.3 (C-24), 15.9 (C-26), 16.1 (C-25), 18.3
(C-6), 19.1 (C-30), 20.7 (C-11), 25.2 (C-12), 26.9 (C-15), 27.4 (C-2),
28.0 (C-23), 29.9 (C-21), 30.2 (C-16), 34.2 (C-7), 35.2 (C-22), 37.1
(C-10), 37.3 (C-13), 38.7 (C-1), 38.8 (C-4), 40.6 (C-8), 42.5 (C-14),
47.6 (C-17), 47.7 (C-19), 48.9 (C-18), 50.3 (C-9), 55.3 (C-5), 59.6
(C-28), 79.0 (C-3), 85.9 (trityl C(Ph)3), 109.3 (C-29), 126.8 (trityl
C-40), 127.7 (trityl C-20, 60), 128.8 (trityl C-30, 50), 144.5 (trityl C-
10), 150.8 (C-20). HRESIMS (positive) m/z 707.4781 [M+Na]+ (calcd
for C49H64O2Na, 707.4804).
Anti-HIV-1 data of conjugates 49, 52 and analogous carboxylic acids 63, 64 against
HIV-1NL4-3 infected MT-4 cellsa
TId
158
63
145
b
c
Compound
EC50
(lM)
IC50 (lM)
49
63
52
64
2
0.045
0.18
7.1
11.3
8.7
5.2.2. General procedure for preparation of 3-O-acyl-28-O-
tritylbetulins (8–10)
0.060
0.310
3-O-Acyl-28-O-tritylbetulins were prepared by refluxing a solu-
tion of 7 (1 equiv), appropriate dicarboxylic acid (4 equiv) or anhy-
dride (4–5 equiv) and DMAP (1–2 equiv) in anhydrous pyridine
(10–20 mL) overnight. After cooling to room temperature, the reac-
tion mixture was diluted with ice-water, and extracted with CHCl3.
The organic layer was washed with water, 2 N HCl solution and
brine, dried over Na2SO4, and concentrated. The crude product
was purified by silica gel column chromatography (hexane/
EtOAc = 10:1).
>14.6
>47
0.096e
0.027
>5e
>52e
>1385
AZT
>37.4
a
b
c
Data presented are averages of at least two separate experiments.
Concentration that inhibits HIV-1NL4-3 replication by 50%.
Concentration that inhibits mock-infected MT-4 cell growth by 50%.
TI = IC50/EC50
Data are taken from Ref. 23.
d
e
.
5.2.2.1. 3-O-(30,30-Dimethylsuccinyl)-28-O-tritylbetulin (8). Yield
38.9% (starting from 4.48 g of 7 and 2,2-dimethylsuccinic acid);
white solid; 3-O-(20,20-dimethylsuccinyl)-28-O-tritylbetulin was
gate compounds were four- and fivefold more active, respectively,
than the corresponding compounds not linked to AZT.
In conclusion, 14 novel conjugates of 3,28-di-O-acyl-betulins
with AZT were prepared as anti-HIV-1 agents. In MT-4 cells, nine
of the conjugates demonstrated potent anti-HIV activity at a sub-
micromolar level with EC50 values ranging from 0.040 to
0.098 lM, and thus, were equipotent or more potent than 2. A
2,2-dimethylsuccinyl group at the C-28 position served as the best
linker between the triterpene scaffold and AZT in this study.
also separated as by-product (11.1% yield). ½a D25
ꢁ
+2.1° (c 1.25,
CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.53 (3H, s, CH3-26), 0.79
(3H, s, CH3-25), 0.81 (3H, s, CH3-24), 0.84 (3H, s, CH3-23), 0.90
(3H, s, CH3-27), 1.30, 1.31 (each 3H, s, dimethylsuccinyl CH3),
1.65 (3H, s, CH3-30), 2.15–2.24 (3H, m, H-22, H-16 and H-19),
2.57, 2.68 (each 1H, d, J = 15.6 Hz, dimethylsuccinyl H2-20), 2.92,
3.15 (each 1H, d, J = 8.8 Hz, H2-28), 4.48 (1H, dd, J = 5.2, 11.2 Hz,
H-3), 4.53, 4.59 (each 1H, br s, H2-29), 7.23 (3H, t, J = 7.2 Hz, trityl
H-40), 7.31 (6H, t, J = 8.0, 7.2 Hz, trityl H-30, 50), 7.50 (6H, d,
J = 8.0 Hz, trityl H-20, 60); 13C NMR (CDCl3, 100 MHz) d 14.7 (C-
27), 15.9 (C-26), 16.1 (C-25), 16.5 (C-24), 18.2 (C-6), 19.1 (C-30),
20.7 (C-11), 23.6 (C-2), 25.1 (C-12), 25.0, 25.6 (dimethylsuccinyl
CH3), 26.9 (C-15), 27.9 (C-23), 29.9 (C-21), 30.1 (C-16), 34.1 (C-7),
35.2 (C-22), 37.0 (C-10), 37.2 (C-13), 37.7 (C-4), 38.3 (C-1), 40.5
(dimethylsuccinyl C-30), 40.6 (C-8), 42.5 (C-14), 44.7 (dim-
ethylsuccinyl C-20), 47.6 (C-17), 47.8 (C-19), 48.9 (C-18), 50.2 (C-
9), 55.3 (C-5), 59.5 (C-28), 81.6 (C-3), 85.8 (trityl C(Ph)3), 109.4
(C-29), 126.8 (trityl C-40), 127.7 (trityl C-20, 60), 128.8 (trityl C-30,
50), 144.5 (trityl C-10), 150.8 (C-20), 170.9 (dimethylsuccinyl
COOꢂ), 182.8 (dimethylsuccinyl COOH). HRESIMS (positive) m/z
835.5286 [M+Na]+ (calcd for C55H72O5Na, 835.5277).
5. Experimental
5.1. General Experimental Procedures
Optical rotations were measured with a JASCO P-2200 digital
polarimeter. NMR spectra (400 MHz for 1H, 100 MHz for 13C, using
TMS as internal standard) were recorded on a Bruker AVANCE 400
spectrometer. HRESIMS was obtained on a Waters LCT Premier.
Column chromatography was performed with silica gel 60 N (63–
210 nm, Merck); Preparative HPLC was carried out on Mightysil
RP-18 GP column (250 ꢀ 20 mm i.d., Kanto Chemical Co., Inc.) or
Cosmosil column (Cholester, 250 ꢀ 20 mm i.d.;
pNAP,
250 ꢀ 20 mm i.d.; Nacalai Tesque). Unless otherwise indicated,
all reagents were purchased from commercial suppliers and used
without further purification.
5.2.2.2. 3-O-(30,30-Dimethylglutaryl)-28-O-tritylbetulin (9). Yield
86.6% (starting from 956 mg of 7 and 3,3-dimethylglutaric anhy-
5.2. Chemical preparation
dride); white solid. ½a D24
ꢁ
+7.5° (c 0.52, CHCl3). 1H NMR (CDCl3,
5.2.1. 28-O-Tritylbetulin (7)
400 MHz) d 0.55 (3H, s, CH3-26), 0.81 (3H, s, CH3-25), 0.85 (3H, s,
CH3-24), 0.86 (3H, s, CH3-23), 0.91 (3H, s, CH3-27), 1.16 (6H, s, dim-
ethylglutaryl CH3), 1.65 (3H, s, CH3-30), 2.15–2.24 (3H, m, H-22, H-
16 and H-19), 2.40, 2.47 (each 1H, d, J = 14.0 Hz, dimethylglutaryl
H2-20), 2.48 (2H, s, dimethylglutaryl H2-40), 2.92, 3.15 (each 1H, d,
J = 8.8 Hz, H2-28), 4.49 (1H, dd, J = 4.8, 10.8 Hz, H-3), 4.54, 4.60
(each 1H, br s, H2-29), 7.23 (3H, t, J = 7.6 Hz, trityl H-40), 7.31
(6H, t, J = 7.6 Hz, trityl H-30, 50), 7.50 (6H, d, J = 7.6 Hz, trityl H-20,
60); 13C NMR (CDCl3, 100 MHz) d 14.7 (C-27), 15.9 (C-26), 16.1
(C-25), 16.5 (C-24), 18.2 (C-6), 19.1 (C-30), 20.8 (C-11), 23.8 (C-
2), 25.2 (C-12), 27.0 (C-15), 27.9, 28.0 (dimethylglutaryl CH3),
28.0 (C-23), 30.0 (C-21), 30.2 (C-16), 32.7 (dimethylglutaryl C-30),
To a solution of 3a (1.02 g, 2.30 mmol) in DMF (15 mL) were
added dimethylaminopyridine (DMAP, 0.33 g, 2.70 mmol) and tri-
phenylmethyl chloride (1.26 g, 4.52 mmol). After refluxing for
6 h, the reaction mixture was poured into ice-water, and extracted
with CHCl3. The organic layer was washed with 2 N HCl solution
and brine, dried over Na2SO4 and concentrated. The obtained resi-
due was chromatographed over silica gel column (hexane/
EtOAc = 10:1) to afford 7 as a white solid (1.16 g, 73.5% yield).
½
a 2D5
ꢁ
ꢂ2.1° (c 1.49, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.54 (3H,
s, CH3-26), 0.77 (3H, s, CH3-24), 0.78 (3H, s, CH3-25), 0.91 (3H, s,
CH3-27), 0.97 (3H, s, CH3-23), 1.65 (3H, s, CH3-30), 2.15–2.24

6456
J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
34.2 (C-7), 35.2 (C-22), 37.1 (C-10), 37.3 (C-13), 37.7 (C-4), 38.4 (C-
1), 40.7 (C-8), 42.5 (C-14), 45.2 (dimethylglutaryl C-40), 45.7 (dim-
ethylglutaryl C-20), 47.6 (C-17), 47.8 (C-19), 49.0 (C-18), 50.3 (C-9),
55.4 (C-5), 59.7 (C-28), 81.5 (C-3), 85.9 (trityl C(Ph)3), 109.4 (C-29),
126.8 (trityl C-40), 127.7 (trityl C-20, 60), 128.8 (trityl C-30, 50), 144.5
(trityl C-10), 150.8 (C-20), 172.3 (dimethylglutaryl COOꢂ), 175.9
(dimethylglutaryl COOH). HRESIMS (positive) m/z 849.5403
[M+Na]+ (calcd for C56H74O5Na, 849.5434).
s, CH3-30), 2.11–2.17 (1H, m, H-21), 2.37–2.45 (2H, m, H-16 and
H-22), 2.61 (1H, dt, J = 5.6, 10.8 Hz, H-19), 2.74, 2.79 (each 1H, d,
J = 16.0 Hz, dimethylglutaryl H2-20), 2.76 (2H, s, dimethylglutaryl
H2-40), 3.65, 4.06 (each 1H, d, J = 10.8 Hz, H2-28), 4.71–4.74 (1H,
m, H-3), 4.74, 4.88 (each 1H, br s, H2-29); 13C NMR (C5D5N,
100 MHz) d 14.9 (C-27), 16.1 (C-26), 16.2 (C-25), 16.9 (C-24),
18.4 (C-6), 19.3 (C-30), 21.0 (C-11), 24.2 (C-2), 25.6 (C-12), 27.5
(C-15), 28.0 (dimethylglutaryl CH3), 28.1 (C-23), 30.0 (C-16), 30.4
(C-21), 32.7 (dimethylglutaryl C-30), 34.4 (C-7), 34.9 (C-22), 37.2
(C-10), 37.5 (C-13), 37.9 (C-4), 38.5 (C-1), 41.2 (C-8), 43.0 (C-14),
45.9 (dimethylglutaryl C-40, 20), 48.3 (C-19), 48.5 (C-17), 49.1 (C-
18), 50.5 (C-9), 55.5 (C-5), 59.4 (C-28), 80.6 (C-3), 109.9 (C-29),
151.2 (C-20), 171.9 (dimethylglutaryl COOꢂ), 174.4 (dimethylglut-
aryl COOH). HRESIMS (positive) m/z 607.4351 [M+Na]+ (calcd for
5.2.2.3. 3-O-Glutaryl-28-O-tritylbetulin (10). Yield 84.1% (starting
from 1.07 g of 7 and glutaric anhydride); white solid. ½a D25
ꢁ
+5.1° (c
1.96, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.53 (3H, s, CH3-26), 0.79
(3H, s, CH3-25), 0.83 (3H, s, CH3-24), 0.83 (3H, s, CH3-23), 0.90 (3H,
s, CH3-27), 1.65 (3H, s, CH3-30), 1.97 (2H, quint, J = 7.2 Hz, glutary
H2-30), 2.15–2.24 (3H, m, H-22, H-16 and H-19), 2.39 (2H, t,
J = 7.2 Hz, glutary H2-20), 2.43 (2H, s, glutaryl H2-40), 2.92, 3.15
(each 1H, d, J = 8.8 Hz, H2-28), 4.48 (1H, dd, J = 5.6, 10.8 Hz, H-3),
4.53, 4.59 (each 1H, br s, H2-29), 7.23 (3H, t, J = 7.2 Hz, trityl H-
40), 7.31 (6H, t, J = 8.0, 7.2 Hz, trityl H-30, 50), 7.50 (6H, d,
J = 8.0 Hz, trityl H-20, 60); 13C NMR (CDCl3, 100 MHz) d 14.7 (C-
27), 15.8 (C-26), 16.1 (C-25), 16.5 (C-24), 18.2 (C-6), 19.1 (C-30),
20.0 (glutaryl C-30), 20.8 (C-11), 23.7 (C-2), 25.1 (C-12), 27.0 (C-
15), 28.0 (C-23), 29.9 (C-21), 30.1 (C-16), 32.9 (glutaryl C-40),
33.6 (glutaryl C-20), 34.1 (C-7), 35.2 (C-22), 37.0 (C-10), 37.2 (C-
13), 37.8 (C-4), 38.3 (C-1), 40.6 (C-8), 42.5 (C-14), 47.6 (C-17),
47.8 (C-19), 48.9 (C-18), 50.2 (C-9), 55.3 (C-5), 59.5 (C-28), 81.1
(C-3), 85.8 (trityl C(Ph)3), 109.4 (C-29), 126.8 (trityl C-40), 127.7
(trityl C-20, 60), 128.8 (trityl C-30, 50), 144.5 (trityl C-10), 150.8 (C-
20), 172.6 (glutaryl COOꢂ), 178.1 (glutaryl COOH). HRESIMS (posi-
tive) m/z 821.5139 [M+Na]+ (calcd for C54H70O5Na, 821.5121).
C37H60O5Na, 607.4338).
5.2.3.3. 3-O-Glutarylbetulin (13). To a solution of 10 (1.50 g,
1.88 mmol) in 95% THF (50 mL) was added anhydrous FeCl3
(1.22 g, 7.52 mmol). The solution was refluxed overnight. After
cooling to room temperature, the solvent was evaporated in vacuo
and the residue was dissolved in CHCl3. The CHCl3 solution was
washed with water and brine, dried over Na2SO4, filtrated and con-
centrated. The crude product was purified by silica column chro-
matography (hexane/EtOAc = 2:1) to give 13 as a white solid
(600 mg, yield 57.4%). ½a D25
ꢁ
+25.1° (c 3.45, CHCl3). 1H NMR (CDCl3,
400 MHz) d 0.84 (3H, s, CH3-24), 0.85 (3H, s, CH3-23), 0.86 (3H, s,
CH3-25), 0.99 (3H, s, CH3-27), 1.03 (3H, s, CH3-26), 1.69 (3H, s,
CH3-30), 1.98 (2H, quint, J = 7.2 Hz, glutary H2-30), 2.35–2.44 (1H,
m, H-19), 2.39 (2H, t, J = 7.2 Hz, glutary H2-20), 2.43 (2H, s, glutaryl
H2-40), 3.35, 3.81 (each 1H, d, J = 10.8 Hz, H2-28), 4.49 (1H, dd,
J = 5.6, 10.4 Hz, H-3), 4.59, 4.69 (each 1H, br s, H2-29); 13C NMR
(CDCl3, 100 MHz) d 14.7 (C-27), 16.0 (C-26), 16.1 (C-25), 16.5 (C-
24), 18.2 (C-6), 19.1 (C-30), 20.1 (glutaryl C-30), 20.9 (C-11), 23.7
(C-2), 25.2 (C-12), 27.1 (C-15), 28.0 (C-23), 29.2 (C-16), 29.8 (C-
21), 32.9 (glutaryl C-40), 33.7 (glutaryl C-20), 34.0 (C-22), 34.2 (C-
7), 37.1 (C-10), 37.3 (C-13), 37.8 (C-4), 38.4 (C-1), 41.0 (C-8), 42.7
(C-14), 47.8 (C-17), 47.8 (C-19), 48.8 (C-18), 50.3 (C-9), 55.4 (C-
5), 60.6 (C-28), 81.1 (C-3), 109.7 (C-29), 150.4 (C-20), 172.6 (gluta-
ryl COOꢂ), 177.6 (glutaryl COOH). HRESIMS (positive) m/z
579.4027 [M+Na]+ (calcd for C35H56O5Na, 579.4025).
5.2.3. General procedure for preparation of 3-O-acylbetulins (11
and 12)
3-O-Acyl-28-O-tritylbetulin (1 equiv) was dissolved in EtOH–
CH2Cl2 and then pyridium p-toluenesulfonate (PPTS) (3–5 equiv)
was added. The reaction mixture was refluxed at 70 °C overnight.
After evaporation under reduced pressure, ice-water was added
to the residue, and extraction was performed with CHCl3. The or-
ganic layer was washed with water and brine, dried over Na2SO4,
and concentrated. The residue was subjected over silica gel column
(hexane/EtOAc = 1:1) to give pure product.
5.2.4. General procedure of hydrogenation
5.2.3.1. 3-O-(30,30-Dimethylsuccinyl)betulin (11). Yield 58.9%
3-O-Acyl-dihydrobetulin derivatives (4, 14, and 15) were pre-
pared by treating a solution of 3-O-acylbetulin derivative (250–
400 mg) in EtOAc–EtOH with 10% palladium catalyst (Pd–C, 240–
800 mg) under hydrogen atmosphere overnight with stirring. The
reaction mixture was filtered, and the filtrate was concentrated un-
der reduced pressure to give a white solid. The purity of the prod-
uct was confirmed by NMR analyses and used in the next reaction
without further purification.
(starting from 1.51 g of 8); white solid. ½a D25
ꢁ
+24.1° (c 1.95, CHCl3).
1H NMR (CDCl3, 400 MHz) d 0.81 (3H, s, CH3-24), 0.84 (6H, s, CH3-
23, 25), 0.97 (3H, s, CH3-27), 1.02 (3H, s, CH3-26), 1.28, 1.30 (each
3H, s, dimethylsuccinyl CH3), 1.73 (3H, s, CH3-30), 2.38 (1H, dt,
J = 5.6, 10.4 Hz, H-19), 2.56, 2.67 (each 1H, d, J = 15.6 Hz, dim-
ethylsuccinyl H2-20), 3.34, 3.80 (each 1H, d, J = 10.8 Hz, H2-28),
4.49 (1H, dd, J = 5.2, 10.4 Hz, H-3), 4.58, 4.68 (each 1H, br s, H2-
29); 13C NMR (CDCl3, 100 MHz) d 14.7 (C-27), 16.0 (C-26), 16.1
(C-25), 16.5 (C-24), 18.2 (C-6), 19.0 (C-30), 20.8 (C-11), 23.6 (C-
2), 25.1 (C-12), 25.0, 25.6 (dimethylsuccinyl CH3), 27.0 (C-15),
27.9 (C-23), 29.1 (C-16), 29.7 (C-21), 33.9 (C-22), 34.1 (C-7), 37.0
(C-10), 37.3 (C-13), 37.7 (C-4), 38.4 (C-1), 40.5 (dimethylsuccinyl
C-30), 40.9 (C-8), 42.7 (C-14), 44.7 (dimethylsuccinyl C-20), 47.7
(C-17), 47.8 (C-19), 48.7 (C-18), 50.3 (C-9), 55.4 (C-5), 60.5 (C-
28), 81.5 (C-3), 109.7 (C-29), 150.4 (C-20), 170.9 (dimethylsuccinyl
COOꢂ), 182.6 (dimethylsuccinyl COOH). HRESIMS (positive) m/z
593.4171 [M+Na]+ (calcd for C36H58O5Na, 593.4182).
5.2.4.1. 3-O-(30,30-Dimethylsuccinyl)dihydrobetulin (14). Yield
100% (starting from 395 mg of 11 and 240 mg of 10% Pd–C); white
solid. ½a 2D4
ꢁ
ꢂ9.3° (c 1.5, CHCl3). 1H NMR (C5D5N, 400 MHz) d 0.79
(6H, s, CH3-25), 0.82, 0.89 (each 2H, d, J = 8.0 Hz, CH3-29, 30),
0.94 (3H, s, CH3-24), 0.96 (3H, s, CH3-23, 26), 0.98 (3H, s, CH3-
27), 1.53 (6H, s, dimethylsuccinyl CH3), 2.38–2.41 (2H, m, H-16
and H-22), 2.88, 2.94 (each 1H, d, J = 15.6 Hz, dimethylsuccinyl
H2-20), 3.58, 4.04 (each 1H, d, J = 10.4 Hz, H2-28), 4.76 (1H, dd,
J = 4.4, 11.6 Hz, H-3); 13C NMR (C5D5N, 100 MHz) d 14.8 (C-27),
15.2 (C-29), 16.1 (C-26), 16.2 (C-25), 16.9 (C-24), 18.4 (C-6), 21.1
(C-11), 22.3 (C-21), 23.2 (C-30), 24.1 (C-2), 25.9, 26.2 (dim-
ethylsuccinyl CH3), 27.2 (C-12), 27.4 (C-15), 28.1 (C-23), 29.8 (C-
20), 30.1 (C-16), 34.5 (C-7), 34.9 (C-22), 37.0 (C-13), 37.2 (C-10),
38.0 (C-4), 38.5 (C-1), 40.9 (dimethylsuccinyl C-30), 41.2 (C-8),
43.1 (C-14), 45.0 (C-19), 45.2 (dimethylsuccinyl C-20), 48.6 (C-17),
5.2.3.2. 3-O-(30,30-Dimethylglutaryl)betulin (12). Yield 67.2%
(starting from 1.0 g of 9); white solid. ½a D25
ꢁ
+24.0° (c 1.88, CHCl3).
1H NMR (C5D5N, 400 MHz) d 0.79 (3H, s, CH3-25), 0.92 (3H, s,
CH3-24), 0.94 (3H, s, CH3-23), 0.96 (3H, s, CH3-26), 1.03 (3H, s,
CH3-27), 1.36, 1.37 (each 3H, s, dimethylglutaryl CH3), 1.76 (3H,

J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
6457
48.4 (C-18), 50.1 (C-9), 55.5 (C-5), 59.4 (C-28), 80.9 (C-3), 171.6
(dimethylsuccinyl COOꢂ), 179.3 (dimethylsuccinyl COOH). HRE-
SIMS (positive) m/z 595.4327 [M+Na]+ (calcd for C36H60O5Na,
595.4338).
38.3 (C-1), 40.6 (dimethylsuccinyl C-30), 40.9 (C-8), 42.6 (C-14),
44.5 (dimethylsuccinyl C-20), 47.7 (C-17), 47.8 (C-19), 48.7 (C-
18), 50.2 (C-9), 55.3 (C-5), 60.4 (C-28), 76.9 (CH(Ph)2), 81.2 (C-3),
109.7 (C-29), 127.0 (benzhydryl C-20, 60), 127.7 (benzyhydryl C-
40), 128.3 (benzhydryl C-30, 50), 140.3 (benzhydryl C-10), 150.4 (C-
20), 170.8 (dimethylsuccinyl 10-COOꢂ), 175.3 (dimethylsuccinyl
40-COOꢂ). HRESIMS (positive) m/z 759.4958 [M+Na]+ (calcd for
5.2.4.2. 3-O-(30,30-Dimethylglutaryl)dihydrobetulin (15). Yield
97.4% (starting from 250 mg of 12 and 250 mg of 10% Pd–C); white
solid. ½a 2D5
ꢁ
ꢂ8.9° (c 1.54, CHCl3). 1H NMR (C5D5N, 400 MHz) d 0.82
C49H68O5Na, 759.4964).
(3H, s, CH3-25), 0.83, 0.90 (each 2H, d, J = 8.0 Hz, CH3-29, 30), 0.93
(3H, s, CH3-24), 0.95 (3H, s, CH3-23), 0.97 (3H, s, CH3-26), 1.00 (3H,
s, CH3-27), 1.37, 1.37 (each 3H, s, dimethylglutaryl CH3), 2.37–2.41
(2H, m, H-16 and H-22), 2.75, 2.79 (each 1H, d, J = 14.4 Hz, dimeth-
ylglutaryl H2-20), 2.77 (2H, s, dimethylglutaryl H2-40), 3.59, 4.05
(each 1H, d, J = 10.8 Hz, H2-28), 4.74 (1H, dd, J = 4.4, 11.6 Hz, H-
3); 13C NMR (C5D5N, 100 MHz) d 14.8 (C-27), 15.2 (C-29), 16.1
(C-26), 16.2 (C-25), 16.9 (C-24), 18.4 (C-6), 21.1 (C-11), 22.3
(C-21), 23.2 (C-30), 24.2 (C-2), 27.2 (C-12), 27.4 (C-15), 28.0 (dim-
ethylglutaryl CH3), 28.1 (C-23), 29.8 (C-20), 30.1 (C-16), 32.7 (dim-
ethylglutaryl C-30), 34.5 (C-7), 34.9 (C-22), 37.0 (C-13), 37.2 (C-10),
37.9 (C-4), 38.5 (C-1), 41.2 (C-8), 43.1 (C-14), 45.0 (C-19), 45.9
(dimethylglutaryl C-40, 20), 48.4 (C-18), 48.6 (C-17), 50.2 (C-9),
55.5 (C-5), 59.4 (C-28), 80.7 (C-3), 172.0 (dimethylglutaryl COOꢂ),
174.4 (dimethylglutaryl COOH). HRESIMS (positive) m/z 609.4493
[M+Na]+ (calcd for C37H62O5Na, 609.4495).
5.2.5.2. 3-O-(30,30-Dimethyglutaryl)betulin benzhydryl ester
(17). Yield 100% (starting from 207 mg of 12); white solid. ½a D27
ꢁ
+18.6° (c 1.29, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.81 (3H, s,
CH3-24), 0.84 (6H, s, CH3-23, 25), 0.99 (3H, s, CH3-27), 1.03 (3H,
s, CH3-26), 1.09 (6H, s, dimethylglutaryl CH3), 1.70 (3H, s, CH3-
30), 2.34–2.45 (1H, m, H-19), 2.36, 2.43 (each 1H, d, J = 14.4 Hz,
dimethylglutaryl H2-20), 2.55, 2.58 (each 1H, d, J = 14.4 Hz, dimeth-
ylglutaryl H2-40), 3.34, 3.81 (each 1H, d, J = 10.8 Hz, H2-28), 4.47
(1H, dd, J = 4.8, 11.2 Hz, H-3), 4.60, 4.70 (each 1H, br s, H2-29),
6.89 (1H, s, CH(Ph)2), 7.25–7.39 (10H, m, aromatic-H); 13C NMR
(CDCl3, 100 MHz) d 14.7 (C-27), 16.0 (C-26), 16.1 (C-25), 16.6 (C-
24), 18.2 (C-6), 19.0 (C-30), 20.8 (C-11), 23.8 (C-2), 25.2 (C-12),
27.0 (C-15), 27.6, 27.7 (dimethylglutaryl CH3), 28.0 (C-23), 29.2
(C-16), 29.7 (C-21), 32.8 (dimethylglutaryl C-30), 33.9 (C-22), 34.1
(C-7), 37.0 (C-10), 37.3 (C-13), 37.7 (C-4), 38.4 (C-1), 40.9 (C-8),
42.7 (C-14), 45.5 (dimethylsuccinyl C-40), 45.8 (dimethylsuccinyl
C-20), 47.8 (C-17), 47.8 (C-19), 48.7 (C-18), 50.3 (C-9), 55.4 (C-5),
60.5 (C-28), 76.7 (CH(Ph)2), 80.9 (C-3), 109.7 (C-29), 127.1 (benz-
hydryl C-20, 60), 127.8 (benzyhydryl C-40), 128.4 (benzhydryl C-30,
50), 140.2, 140.3 (benzhydryl C-10), 150.4 (C-20), 170.8 (dimethyl-
glutaryl 50-COOꢂ), 171.6 (dimethylglutaryl 10-COOꢂ). HRESIMS
(positive) m/z 773.5124 [M+Na]+ (calcd for C50H70O5Na, 773.5121).
5.2.4.3. 3-O-Glutaryl-dihydrobetulin (4). Yield 91.1% (starting
from 387 mg of 13 and 800 mg of 10% Pd–C); white solid. ½a D25
ꢁ
ꢂ5.4° (c 2.40, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.78, 0.85 (each
2H, d, J = 8.0 Hz, CH3-29, 30), 0.85 (3H, s, CH3-24), 0.85 (3H, s, CH3-
23), 0.87 (3H, s, CH3-25), 0.96 (3H, s, CH3-27), 1.04 (3H, s, CH3-26),
1.99 (2H, quint, J = 7.2 Hz, glutary H2-30), 2.40 (2H, t, J = 7.2 Hz, glu-
tary H2-20), 2.43 (2H, s, glutaryl H2-40), 3.32, 3.79 (each 1H, d,
J = 10.8 Hz, H2-28), 4.50 (1H, dd, J = 6.4, 10.0 Hz, H-3); 13C NMR
(CDCl3, 100 MHz) d 14.6 (C-27), 14.9 (C-29), 16.0 (C-26), 16.1 (C-
25), 16.6 (C-24), 18.2 (C-6), 20.1 (glutaryl C-30), 20.9 (C-11), 21.7
(C-21), 22.9 (C-30), 23.7 (C-2), 26.9 (C-12), 26.9 (C-15), 28.0 (C-
23), 29.3 (C-16), 29.5 (C-20), 33.0 (glutaryl C-40), 33.7 (glutaryl C-
20), 34.0 (C-22), 34.3 (C-7), 36.9 (C-13), 37.1 (C-10), 37.8 (C-4),
38.4 (C-1), 41.0 (C-8), 42.9 (C-14), 44.6 (C-19), 47.9 (C-17), 48.1
(C-18), 50.0 (C-9), 55.4 (C-5), 60.6 (C-28), 81.1 (C-3), 172.7 (glutaryl
COOꢂ), 177.9 (glutaryl COOH). HRESIMS (positive) m/z 581.4180
[M+Na]+ (calcd for C35H58O5Na, 581.4182).
5.2.5.3. 3-O-(30,30-Dimethylsuccinyl)dihydrobetulin benzhydryl
ester (18). Yield 100% (starting from 400 mg of 14); white solid.
½
a 2D7
ꢁ
ꢂ7.9° (c 5.27, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.75 (3H,
s, CH3-24), 0.79 (3H, s, CH3-23), 0.78, 0.85 (each 2H, d, J = 8.0 Hz,
CH3-29, 30), 0.81 (3H, s, CH3-25), 0.96 (3H, s, CH3-27), 1.03 (3H,
s, CH3-26), 1.31, 1.32 (each 3H, s, dimethylsuccinyl CH3), 2.66,
2.70 (each 1H, d, J = 15.6 Hz, dimethylsuccinyl H2-20), 3.31, 3.78
(each 1H, d, J = 10.8 Hz, H2-28), 4.45 (1H, dd, J = 5.0, 10.8 Hz, H-
3), 6.87 (1H, s, CH(Ph)2), 7.27–7.33 (10H, m, aromatic-H); 13C
NMR (CDCl3, 100 MHz) d 14.6 (C-27), 14.9 (C-29), 15.9 (C-26),
16.0 (C-25), 16.5 (C-24), 18.1 (C-6), 20.8 (C-11), 21.7 (C-21), 22.9
(C-30), 23.5 (C-2), 25.2, 25.4 (dimethylsuccinyl CH3), 26.8 (C-12),
26.9 (C-15), 27.9 (C-23), 29.3 (C-16), 29.5 (C-20), 34.0 (C-22),
34.2 (C-7), 36.8 (C-13), 37.0 (C-10), 37.6 (C-4), 38.3 (C-1), 40.7
(dimethylsuccinyl C-30), 40.9 (C-8), 42.8 (C-14), 44.5 (C-19 and
dimethylsuccinyl C-20), 47.9 (C-17), 48.0 (C-18), 49.9 (C-9), 55.3
(C-5), 60.6 (C-28), 77.0 (CH(Ph)2), 81.3 (C-3), 127.0 (benzhydryl
C-20, 60), 127.7 (benzyhydryl C-40), 128.4 (benzhydryl C-30, 50),
140.3 (benzhydryl C-10), 170.8 (dimethylsuccinyl 10-COOꢂ), 175.4
(dimethylsuccinyl 40-COOꢂ). HRESIMS (positive) m/z 761.5118
[M+Na]+ (calcd for C49H70O5Na, 761.5121).
5.2.5. General procedure for syntheses of 3-O-acylbetulin
benzhydryl ester derivatives (16–20)
To a solution of 3-O-acylbetulin derivatives (1 equiv) in CH2Cl2–
CH3OH, freshly prepared diphenyldiazomethane was added until
the reaction solution turned pink. The resulting mixture was kept
stirring at room temperature overnight. After evaporated solvent
in vacuo, the residue was subjected over silica gel column (hex-
ane/EtOAc = 10:1) to give the desired product.
5.2.5.1. 3-O-(30,30-Dimethylsuccinyl)betulin benzhydryl ester
(16). Yield 100% (starting from 265 mg of 11); white solid. ½a D17
ꢁ
+16.9° (c 1.74, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.75 (3H, s,
CH3-24), 0.79 (3H, s, CH3-23), 0.80 (3H, s, CH3-25), 0.99 (3H, s,
CH3-27), 1.03 (3H, s, CH3-26), 1.32, 1.32 (each 3H, s, dimethylsucci-
nyl CH3), 1.70 (3H, s, CH3-30), 2.40 (1H, dt, J = 5.6, 10.4 Hz, H-19),
2.66, 2.70 (each 1H, d, J = 16.0 Hz, dimethylsuccinyl H2-20), 3.34,
3.80 (each 1H, d, J = 10.8 Hz, H2-28), 4.45 (1H, dd, J = 5.2, 10.8 Hz,
H-3), 4.60, 4.70 (each 1H, br s, H2-29), 6.87 (1H, s, CH(Ph)2),
7.25–7.34 (10H, m, aromatic-H); 13C NMR (CDCl3, 100 MHz) d
14.7 (C-27), 15.9 (C-26), 16.0 (C-25), 16.4 (C-24), 18.1 (C-6), 19.0
(C-30), 20.8 (C-11), 23.5 (C-2), 25.1 (C-12), 25.1, 25.4 (dim-
ethylsuccinyl CH3), 27.0 (C-15), 27.8 (C-23), 29.1 (C-16), 29.7 (C-
21), 33.9 (C-22), 34.1 (C-7), 37.0 (C-10), 37.2 (C-13), 37.6 (C-4),
5.2.5.4. 3-O-(30,30-Dimethylglutaryl)dihydrobetulin benzhydryl
ester (19). Yield 85.3% (starting from 228 mg of 15); white solid.
½
a 2D4
ꢁ
ꢂ7.7° (c 1.26, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.79 (3H,
s, CH3-24), 0.82 (3H, s, CH3-23, 25), 0.76, 0.83 (each 2H, d,
J = 8.0 Hz, CH3-29, 30), 0.94 (3H, s, CH3-27), 1.00 (3H, s, CH3-26),
1.07 (6H, s, dimethylglutaryl CH3), 2.34, 2.41 (each 1H, d,
J = 14.4 Hz, dimethylglutaryl H2-20), 2.53, 2.56 (each 1H, d,
J = 14.4 Hz, dimethylglutaryl H2-40), 3.30, 3.76 (each 1H, d,
J = 10.8 Hz, H2-28), 4.46 (1H, dd, J = 4.8, 10.8 Hz, H-3), 6.87 (1H, s,
CH(Ph)2), 7.23–7.34 (10H, m, aromatic-H); 13C NMR (CDCl3,
100 MHz) d 14.5 (C-27), 14.8 (C-29), 15.9 (C-26), 16.0 (C-25),
16.5 (C-24), 18.1 (C-6), 20.7 (C-11), 21.6 (C-21), 22.9 (C-30), 23.7

6458
J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
(C-2), 26.8 (C-12), 26.8 (C-15), 27.5, 27.6 (dimethylglutaryl CH3),
27.9 (C-23), 29.3 (C-16), 29.4 (C-20), 32.7 (dimethylglutaryl C-30),
34.0 (C-22), 34.1 (C-7), 36.7 (C-13), 36.9 (C-10), 37.6 (C-4), 38.3
(C-1), 40.9 (C-8), 42.8 (C-14), 44.5 (C-19), 45.3 (dimethylglutaryl
C-40), 45.7 (dimethylglutaryl C-20), 47.8 (C-17), 48.0 (C-18), 49.9
(C-9), 55.2 (C-5), 60.1 (C-28), 76.6 (CH(Ph)2), 80.8 (C-3), 127.0
(benzhydryl C-20, 60), 127.7 (benzyhydryl C-40), 128.3 (benzhydryl
C-30, 50), 140.2 (benzhydryl C-10), 170.7 (dimethylglutaryl 50-
COOꢂ), 171.5 (dimethylglutaryl 10-COOꢂ). HRESIMS (positive) m/
z 775.5303 [M+Na]+ (calcd for C50H72O5Na, 775.5277).
10), 150.1 (C-20), 170.9 (3-O-dimethylsuccinyl 10-COOꢂ), 172.5
(28-O-succinyl 100-COOꢂ), 175.4 (3-O-dimethylsuccinyl 40-COOꢂ),
176.8 (28-O-succinyl COOH). HRESIMS (positive) m/z 859.5112
[M+Na]+ (calcd for C53H72O8Na, 859.5125).
5.2.6.2. 3-O-(40-Benzhydryloxy-30,30-dimethylsuccinyl)-28-O-(300,
300-dimethylsuccinyl)betulin (22) and 3-O-(40-benzhydryloxy-
30,30-dimethylsuccinyl)-28-O-(200,200-dimethylsuccinyl)betulin
(23). Yield 68.4% and 31.6%, respectively (starting from 114 mg of
16 and 2,2-dimethylsuccinic acid); separated by preparative HPLC
(pNAP, MeOH/2% HOAc = 97:3) for spectroscopic analysis. For syn-
5.2.5.5. 3-O-Glutaryl-dihydrobetulin benzhydryl ester (20). Yield
thesis, mixture was used for next reaction without purification.
96.4% (starting from 332 mg of 4); white solid. ½a D27
ꢁ
ꢂ4.2° (c 3.84,
Compound 22: ½a 2D6
ꢁ
+6.0° (c 6.81, CHCl3). 1H NMR (CDCl3,
CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.84 (3H, s, CH3-24), 0.85
(3H, s, CH3-23), 0.79, 0.85 (each 2H, d, J = 8.0 Hz, CH3-29, 30),
0.87 (3H, s, CH3-25), 0.97 (3H, s, CH3-27), 1.03 (3H, s, CH3-26),
2.00 (2H, t, J = 7.2 Hz, glutaryl H2-30), 2.36 (2H, t, J = 7.2 Hz, glutaryl
H2-20), 2.51 (2H, t, J = 7.2 Hz, glutaryl H2-40), 3.32, 3.78 (each 1H, d,
J = 10.8 Hz, H2-28), 4.50 (1H, dd, J = 5.6, 10.4 Hz, H-3), 6.90 (1H, s,
CH(Ph)2), 7.26–7.34 (10H, m, aromatic-H); 13C NMR (CDCl3,
100 MHz) d 14.6 (C-27), 14.9 (C-29), 16.0 (C-26), 16.1 (C-25),
16.5 (C-24), 18.2 (C-6), 20.4 (glutaryl C-30), 20.9 (C-11), 21.7 (C-
21), 22.9 (C-30), 23.7 (C-2), 26.9 (C-12), 26.9 (C-15), 28.0 (C-23),
29.3 (C-16), 29.5 (C-20), 33.6 (glutaryl C-40), 33.7 (glutaryl C-20),
34.0 (C-22), 34.3 (C-7), 36.8 (C-13), 37.1 (C-10), 37.8 (C-4), 38.4
(C-1), 41.0 (C-8), 42.9 (C-14), 44.6 (C-19), 47.9 (C-17), 48.1 (C-
18), 50.0 (C-9), 55.3 (C-5), 60.6 (C-28), 76.9 (CH(Ph)2), 81.0 (C-3),
127.1 (benzhydryl C-20, 60), 127.9 (benzyhydryl C-40), 128.5 (benz-
hydryl C-30, 50), 140.2 (benzhydryl C-10), 171.9 (glutaryl 50-COOꢂ),
172.6 (glutaryl 10-COOꢂ). HRESIMS (positive) m/z 747.4962
[M+Na]+ (calcd for C48H68O5Na, 747.4964).
400 MHz) d 0.75 (3H, s, CH3-24), 0.79 (3H, s, CH3-23), 0.80 (3H, s,
CH3-25), 0.97 (3H, s, CH3-27), 1.03 (3H, s, CH3-26), 1.32 (12H, s,
dimethylsuccinyl CH3), 1.70 (3H, s, CH3-30), 2.43 (1H, dt, J = 5.6,
10.8 Hz, H-19), 2.64, 2.70 (each 1H, d, J = 16.0 Hz, 3-O-dim-
ethylsuccinyl H2-20), 2.66 (2H, s, 28-O-dimethylsuccinyl H2-200),
3.88, 4.29 (each 1H, d, J = 10.8 Hz, H2-28), 4.45 (1H, dd, J = 5.2,
10.8 Hz, H-3), 4.61, 4.70 (each 1H, br s, H2-29), 6.87 (1H, s,
CH(Ph)2), 7.25–7.33 (10H, m, aromatic-H); 13C NMR (CDCl3,
100 MHz) d 14.8 (C-27), 16.1 (C-26), 16.1 (C-25), 16.5 (C-24),
18.2 (C-6), 19.2 (C-30), 20.9 (C-11), 23.6 (C-2), 25.2 (C-12), 25.2,
25.4 (3-O-dimethylsuccinyl CH3), 25.3, 25.4 (28-O-dimethylsucci-
nyl CH3), 27.0 (C-15), 27.9 (C-23), 29.6 (C-21), 29.8 (C-16), 34.2
(C-7), 34.6 (C-22), 37.1 (C-10), 37.7 (C-4, C-13), 38.4 (C-1), 40.7
(dimethylsuccinyl C-30, C-300), 40.9 (C-8), 42.7 (C-14), 44.4 (28-O-
dimethylsuccinyl C-200), 44.6 (3-O-dimethylsuccinyl C-20), 46.3
(C-17), 47.7 (C-19), 48.9 (C-18), 50.3 (C-9), 55.4 (C-5), 63.1 (C-
28), 77.0 (CH(Ph)2), 81.3 (C-3), 109.8 (C-29), 127.0, 127.1 (benzhy-
dryl C-20, 60), 127.7 (benzyhydryl C-40), 128.4 (benzhydryl C-30, 50),
140.4 (benzhydryl C-10), 150.1 (C-20), 170.8 (3-O-dimethylsuccinyl
10-COOꢂ), 171.5 (28-O-dimethylsuccinyl 100-COOꢂ), 175.4 (3-O-
dimethylsuccinyl 40-COOꢂ), 182.4 (28-O-dimethylsuccinyl COOH).
HRESIMS (positive) m/z 887.5421 [M+Na]+ (calcd for C55H76O8Na,
887.5438).
5.2.6. General procedure for syntheses of 3,28-di-O-acylbetulin
derivatives (21–34)
3,28-Di-O-acylbetulin derivatives were prepared by refluxing a
solution of 3-O-acylbetulin benzhydryl ester derivatives (1 equiv),
DMAP (1.5–4.5 equiv) and appropriate dicarboxylic acid (6–
9 equiv) or anhydride (3–6 equiv) in anhydrous pyridine (5–
8 mL) overnight. After cooling to room temperature, the reaction
mixture was poured into ice-water, and extracted with CHCl3.
The organic layer was washed with water, 2 N HCl solution and
brine in turn, dried over Na2SO4, and concentrated. The residue
was chromatographed over silica gel column (CHCl3 or hexane/
EtOAc = 5:1) or purified by preparative HPLC.
Compound 23: ½a 2D7
ꢁ
+6.3° (c 3.15, CHCl3). 1H NMR (CDCl3,
400 MHz) d 0.75 (3H, s, CH3-24), 0.79 (3H, s, CH3-23), 0.80 (3H, s,
CH3-25), 0.98 (3H, s, CH3-27), 1.03 (3H, s, CH3-26), 1.31, 1.32 (each
3H, s, 3-O-dimethylsuccinyl CH3), 1.32 (6H, s, 28-O-dimethylsucci-
nyl CH3), 1.70 (3H, s, CH3-30), 2.44 (1H, dt, J = 5.6, 10.8 Hz, H-19),
2.64, 2.70 (each 1H, d, J = 15.6 Hz, 3-O-dimethylsuccinyl H2-20),
2.66 (2H, s, 28-O-dimethylsuccinyl H2-300), 3.85, 4.32 (each 1H, d,
J = 10.8 Hz, H2-28), 4.45 (1H, dd, J = 5.2, 10.8 Hz, H-3), 4.61, 4.70
(each 1H, br s, H2-29), 6.87 (1H, s, CH(Ph)2), 7.24–7.32 (10H, m,
aromatic-H); 13C NMR (CDCl3, 100 MHz) d 14.8 (C-27), 16.0 (C-
26), 16.1 (C-25), 16.5 (C-24), 18.2 (C-6), 19.2 (C-30), 20.8 (C-11),
23.6 (C-2), 25.2 (C-12), 25.2, 25.4 (3-O-dimethylsuccinyl CH3),
25.3, 25.5 (28-O-dimethylsuccinyl CH3), 27.0 (C-15), 27.9 (C-23),
29.6 (C-21), 29.9 (C-16), 34.1 (C-7), 34.5 (C-22), 37.1 (C-10), 37.7
(C-4, 13), 38.4 (C-1), 40.7 (dimethylsuccinyl C-30, C-200), 40.9 (C-
8), 42.7 (C-14), 44.0 (28-O-dimethylsuccinyl C-300), 44.6 (3-O-dim-
ethylsuccinyl C-20), 46.6 (C-17), 47.8 (C-19), 48.9 (C-18), 50.3 (C-9),
55.4 (C-5), 63.3 (C-28), 77.0 (CH(Ph)2), 81.3 (C-3), 109.8 (C-29),
127.0, 127.1 (benzhydryl C-20, 60), 127.7 (benzyhydryl C-40),
128.4 (benzhydryl C-30, 50), 140.4 (benzhydryl C-10), 150.1 (C-20),
170.8 (3-O-dimethylsuccinyl 10-COOꢂ), 175.4 (3-O-dimethylsucci-
nyl 40-COOꢂ, 28-O-dimethylsuccinyl COOH), 177.0 (28-O-dim-
ethylsuccinyl 100-COO–. HRESIMS (positive) m/z 887.5440
[M+Na]+ (calcd for C55H76O8Na, 887.5438).
5.2.6.1.
3-O-(40-Benzhydryloxy-30,30-dimethylsuccinyl)-28-O-
succinylbetulin (21). Yield 76.8% (starting from 504.6 mg of 16
and succinic anhydride); white solid. ½a D18
ꢁ
+5.6° (c 0.5, CHCl3). 1H
NMR (CDCl3, 500 MHz) d 0.73 (3H, s, CH3-24), 0.77 (6H, s, CH3-23
and CH3-25), 0.96 (3H, s, CH3-27), 1.01 (3H, s, CH3-26), 1.30, 1.31
(each 3H, s, dimethylsuccinyl CH3), 1.68 (3H, s, CH3-30), 2.43
(1H, dt, J = 5.5, 10.9 Hz, H-19), 2.64, 2.69 (each 1H, d, J = 15.8 Hz,
3-O-dimethylsuccinyl H2-20), 2.68 (4H, t, J = 7.3 Hz, 28-O-succinyl
H2-200, H2-300), 3.88, 4.30 (each 1H, d, J = 10.8 Hz, H2-28), 4.43
(1H, dd, J = 5.2, 11.2 Hz, H-3), 4.59, 4.68 (each 1H, br s, H2-29),
6.85 (1H, s, CH(Ph)2), 7.24–7.32 (10H, m, aromatic-H); 13C NMR
(CDCl3, 125 MHz) d 14.7 (C-27), 16.0 (C-26), 16.1 (C-25), 16.5 (C-
24), 18.1 (C-6), 19.1 (C-30), 20.7 (C-11), 23.5 (C-2), 25.1 (C-12),
25.2, 25.4 (3-O-dimethylsuccinyl CH3), 27.0 (C-15), 27.9 (C-23),
28.8 (28-O-succinyl C-300), 29.0 (28-O-succinyl C-200), 29.5 (C-21),
29.7 (C-16), 34.0 (C-7), 34.5 (C-22), 37.0 (C-10), 37.5 (C-13), 37.6
(C-4), 38.3 (C-1), 40.7 (dimethylsuccinyl C-30), 40.8 (C-8), 42.6 (C-
14), 44.5 (3-O-dimethylsuccinyl C-20), 46.4 (C-17), 47.7 (C-19),
48.7 (C-18), 50.2 (C-9), 55.3 (C-5), 63.2 (C-28), 77.0 (CH(Ph)2),
81.3 (C-3), 109.9 (C-29), 127.0 (benzhydryl C-20, 60), 127.7 (ben-
zyhydryl C-40), 128.4 (benzhydryl C-30, 50), 140.3 (benzhydryl C-
5.2.6.3.
3-O-(40-Benzhydryloxy-30,30-dimethylsuccinyl)-28-O-
glutarylbetulin (24). Yield 59.5% (starting from 406 mg of 16
and glutaric anhydride); white solid. ½a D15
ꢁ
+10.8° (c 0.5, CHCl3).
1H NMR (CDCl3, 500 MHz) d 0.73 (3H, s, CH3-24), 0.77 (6H, s,
CH3-23 and CH3-25), 0.96 (3H, s, CH3-27), 1.01 (3H, s, CH3-26),

J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
6459
1.30, 1.31 (each 3H, s, dimethylsuccinyl CH3), 1.68 (3H, s, CH3-30),
1.97 (2H, quint, J = 7.2 Hz, 28-O-glutaryl H2-300), 2.41–2.46 (1H, m,
J = 5.5, 10.9 Hz, H-19), 2.43 (2H, t, J = 7.2 Hz, 28-O-glutaryl H2-200),
2.44 (2H, t, J = 7.2 Hz, 28-O-glutaryl H2-400), 2.64, 2.69 (each 1H,
d, J = 15.8 Hz, 3-O-dimethylsuccinyl H2-20), 3.85, 4.28 (each 1H, d,
J = 10.8 Hz, H2-28), 4.43 (1H, dd, J = 5.2, 11.4 Hz, H-3), 4.59, 4.68
(each 1H, br s, H2-29), 6.85 (1H, s, CH(Ph)2), 7.24–7.32 (10H, m,
aromatic-H); 13C NMR (CDCl3, 125 MHz) d 14.7 (C-27), 16.0
(C-26), 16.1 (C-25), 16.5 (C-24), 18.1 (C-6), 19.1 (C-30), 20.0 (28-
O-glutaryl C-300), 20.8 (C-11), 23.5 (C-2), 25.1 (C-12), 25.2, 25.4
(3-O-dimethylsuccinyl CH3), 27.0 (C-15), 27.9 (C-23), 29.5 (C-21),
29.7 (C-16), 32.8 (28-O-glutaryl C-400), 33.3 (28-O-glutaryl C-200),
34.1 (C-7), 34.5 (C-22), 37.0 (C-10), 37.6 (2C, C-13 and C-4), 38.4
(C-1), 40.7 (3-O-dimethylsuccinyl C-30), 40.9 (C-8), 42.7 (C-14),
44.5 (3-O-dimethylsuccinyl C-20), 46.4 (C-17), 47.7 (C-19), 48.8
(C-18), 50.2 (C-9), 55.3 (C-5), 62.8 (C-28), 77.0 (CH(Ph)2), 81.3 (C-
3), 109.9 (C-29), 127.0 (benzhydryl C-20, 60), 127.7 (benzyhydryl
C-40), 128.4 (benzhydryl C-30, 50), 140.3 (benzhydryl C-10), 150.1
(C-20), 170.9 (3-O-dimethylsuccinyl 10-COOꢂ), 173.3 (28-O-gluta-
ryl 100-COOꢂ), 175.4 (3-O-dimethylsuccinyl 40-COOꢂ), 177.7 (28-
O-glutaryl COOH). HRESIMS (positive) m/z 873.5286 [M+Na]+
(calcd for C54H74O8Na, 873.5281).
(C-15), 27.6, 27.7 (3-O-dimethylglutaryl CH3), 27.8 (28-O-dimeth-
ylglutaryl CH3), 28.0 (C-23), 29.5 (C-21), 29.8 (C-16), 32.6 (28-O-
dimethylglutaryl C-300), 32.8 (28-O-dimethylglutaryl C-30), 34.1
(C-7), 34.6 (C-22), 37.0 (C-10), 37.6 (C-4, 13), 38.4 (C-1), 40.9
(C-8), 42.7 (C-14), 45.1 (28-O-dimethylglutaryl C-400), 45.3 (28-O-
dimethylglutaryl C-200), 45.4 (3-O-dimethylglutaryl C-40), 45.8 (3-
O-dimethylglutaryl C-20), 46.2 (C-17), 47.7 (C-19), 48.8 (C-18),
50.2 (C-9), 55.4 (C-5), 62.9 (C-28), 76.7 (CH(Ph)2), 80.9 (C-3),
109.9 (C-29), 127.1 (benzhydryl C-20, 60), 127.8 (benzyhydryl
C-40), 128.4 (benzhydryl C-30, 50), 140.2, 140.3 (benzhydryl C-10),
150.0 (C-20), 170.8 (3-O-dimethylglutaryl 50-COOꢂ), 171.7
(3-O-dimethylglutaryl 10-COOꢂ), 172.8 (28-O-dimethylglutaryl
100-COOꢂ), 176.5 (28-O-dimethylglutaryl COOH). HRESIMS (posi-
tive) m/z 915.5771 [M+Na]+ (calcd for C57H80O8Na, 915.5751).
5.2.6.6. 3-O-(40-Benzhydryloxy-30,30-dimethylsuccinyl)-28-O-(300,
300-dimethylsuccinyl)dihydrobetulin (27) and 3-O-(40-benzhyd-
ryloxy-30,30-dimethylsuccinyl)-28-O-(200,200-dimethylsuccinyl)-
dihydrobetulin (28). Total yield 96.3% (starting from 330 mg of 18
and 2,2-dimethylsuccinic acid); used in next reaction without puri-
fication. For spectroscopic analysis, a small amount of sample was
further purified by preparative HPLC
HOAc = 97:3); white solid.
(pNAP, MeOH/2%
5.2.6.4.
3-O-(40-Benzhydryloxy-30,30-dimethylsuccinyl)-28-O-
Compound 27: ½a 2D6
ꢁ
ꢂ12.3° (c 3.50, CHCl3). 1H NMR (CDCl3,
(300,300-dimethylglutaryl)betulin (25). Yield 87.0% (starting from
400 MHz) d 0.75 (3H, s, CH3-24), 0.77, 0.85 (each 3H, d, J = 8.0 Hz,
CH3-29, 30), 0.79 (3H, s, CH3-23), 0.80 (3H, s, CH3-25), 0.94 (3H,
s, CH3-27), 1.03 (3H, s, CH3-26), 1.31, 1.32 (each 3H, s, 3-O-dim-
ethylsuccinyl CH3), 1.31 (6H, s, 28-O-dimethylsuccinyl CH3), 2.65
(2H, s, 28-O-dimethylsuccinyl H2-200), 2.66, 2.70 (each 1H, d,
J = 15.6 Hz, 3-O-dimethylsuccinyl H2-20), 3.85, 4.26 (each 1H, d,
J = 11.2 Hz, H2-28), 4.45 (1H, dd, J = 5.2, 10.8 Hz, H-3), 6.87 (1H, s,
CH(Ph)2), 7.25–7.33 (10H, m, aromatic-H); 13C NMR (CDCl3,
100 MHz) d 14.6 (C-27), 14.9 (C-29), 16.0 (C-25, C-26), 16.5 (C-
24), 18.1 (C-6), 20.8 (C-11), 21.5 (C-21), 22.9 (C-30), 23.5 (C-2),
25.2, 25.4 (3-O-dimethylsuccinyl CH3), 25.3, 25.4 (28-O-dim-
ethylsuccinyl CH3), 26.8 (C-12, C-15), 27.9 (C-23), 29.4 (C-20),
29.8 (C-16), 34.1 (C-7), 34.6 (C-22), 36.9 (C-10), 37.1 (C-13), 37.6
(C-4), 38.3 (C-1), 40.5 (28-O-dimethylsuccinyl C-300), 40.7 (3-O-
dimethylsuccinyl C-30), 40.9 (C-8), 42.8 (C-14), 44.4 (28-O-dim-
ethylsuccinyl C-200), 44.5 (C-19, 3-O-dimethylsuccinyl C-20), 46.4
(C-17), 48.1 (C-18), 49.9 (C-9), 55.3 (C-5), 63.1 (C-28), 77.0
(CH(Ph)2), 81.3 (C-3), 127.0, 127.1 (benzhydryl C-20, 60), 127.7
(benzyhydryl C-40), 128.4 (benzhydryl C-30, 50), 140.3 (benzhydryl
C-10), 170.9 (3-O-dimethylsuccinyl 10-COOꢂ), 171.6 (28-O-dim-
ethylsuccinyl 100-COOꢂ), 175.4 (3-O-dimethylsuccinyl 40-COOꢂ),
182.9 (28-O-dimethylsuccinyl COOH). HRESIMS (positive) m/z
889.5585 [M+Na]+ (calcd for C55H78O8Na, 889.5594).
222 mg of 16 and 3,3-dimethylglutaric acid); white solid. ½a D25
ꢁ
+6.8° (c 3.59, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.74 (3H, s,
CH3-24), 0.79 (3H, s, CH3-23), 0.79 (3H, s, CH3-25), 0.98 (3H, s,
CH3-27), 1.03 (3H, s, CH3-26), 1.16 (6H, s, dimethylglutaryl CH3),
1.31, 1.32 (each 3H, s, dimethylsuccinyl CH3), 1.70 (3H, s,
CH3-30), 2.40–2.50 (1H, m, H-19), 2.47, 2.48 (each 2H, s, dimethyl-
glutaryl H2-200, 400), 2.65, 2.70 (each 1H, d, J = 16.0 Hz, 3-O-dim-
ethylsuccinyl H2-20), 3.86, 4.29 (each 1H, d, J = 10.8 Hz, H2-28),
4.44 (1H, dd, J = 5.2, 10.8 Hz, H-3), 4.60, 4.70 (each 1H, br s, H2-
29), 6.87 (1H, s, CH(Ph)2), 7.24–7.33 (10H, m, aromatic-H); 13C
NMR (CDCl3, 100 MHz) d 14.7 (C-27), 16.0 (C-26), 16.1 (C-25),
16.5 (C-24), 18.1 (C-6), 19.1 (C-30), 20.8 (C-11), 23.5 (C-2), 25.2
(C-12), 25.2, 25.4 (3-O-dimethylsuccinyl CH3), 26.9 (C-15), 27.8
(28-O-dimethylsuccinyl CH3), 27.9 (C-23), 29.5 (C-21), 29.8 (C-
16), 32.6 (dimethylglutaryl C-300), 34.0 (C-7), 34.6 (C-22), 37.0 (C-
10), 37.6 (C-4, 13), 38.3 (C-1), 40.7 (dimethylsuccinyl C-30), 40.8
(C-8), 42.6 (C-14), 44.5 (3-O-dimethylsuccinyl C-20), 45.1 (dimeth-
ylglutaryl C-400), 45.3 (dimethylglutaryl C-200), 46.2 (C-17), 47.7 (C-
19), 48.8 (C-18), 50.2 (C-9), 55.3 (C-5), 62.8 (C-28), 77.0 (CH(Ph)2),
81.3 (C-3), 109.9 (C-29), 127.0 (benzhydryl C-20, 60), 127.7 (ben-
zyhydryl C-40), 128.4 (benzhydryl C-30, 50), 140.3 (benzhydryl C-
10), 150.0 (C-20), 170.9 (3-O-dimethylsuccinyl 10-COOꢂ), 172.7
(28-O-dimethylglutaryl 100-COOꢂ), 175.4 (3-O-dimethylsuccinyl
40-COOꢂ), 176.9 (28-O-dimethylglutaryl COOH). HRESIMS (posi-
tive) m/z 901.5583 [M+Na]+ (calcd for C56H78O8Na, 901.5594).
Compound 28: ½a 2D7
ꢁ
ꢂ11.4° (c 2.30, CHCl3). 1H NMR (CDCl3,
400 MHz) d 0.75 (3H, s, CH3-24), 0.78, 0.85 (each 3H, d, J = 8.0 Hz,
CH3-29, 30), 0.79 (3H, s, CH3-23), 0.80 (3H, s, CH3-25), 0.95 (3H,
s, CH3-27), 1.03 (3H, s, CH3-26), 1.32, 1.31 (each 3H, s, 3-O-dim-
ethylsuccinyl CH3), 1.31 (each 3H, s, 28-O-dimethylsuccinyl CH3),
2.66, 2.70 (each 1H, d, J = 15.6 Hz, 3-O-dimethylsuccinyl H2-20),
2.65 (2H, s, 28-O-dimethylsuccinyl H2-300), 3.79, 4.31 (each 1H, d,
J = 10.8 Hz, H2-28), 4.45 (1H, dd, J = 5.2, 11.2 Hz, H-3), 6.87 (1H, s,
CH(Ph)2), 7.25–7.33 (10H, m, aromatic-H); 13C NMR (CDCl3,
100 MHz) d 14.6 (C-27), 14.9 (C-29), 16.0 (C-25, C-26), 16.5 (C-
24), 18.1 (C-6), 20.8 (C-11), 21.5 (C-21), 22.9 (C-30), 23.5 (C-2),
25.2, 25.4 (3-O-dimethylsuccinyl CH3), 25.4, 25.5 (28-O-dim-
ethylsuccinyl CH3), 26.8 (C-12, C-15), 27.9 (C-23), 29.4 (C-20),
29.9 (C-16), 34.1 (C-7), 34.7 (C-22), 37.0 (C-10), 37.1 (C-13), 37.7
(C-4), 38.4 (C-1), 40.7 (3-O-dimethylsuccinyl C-30, 28-O-dim-
ethylsuccinyl 200), 40.9 (C-8), 42.8 (C-14), 43.9 (28-O-dim-
ethylsuccinyl C-300), 44.5 (C-19, 3-O-dimethylsuccinyl C-20), 46.6
(C-17), 48.2 (C-18), 49.9 (C-9), 55.3 (C-5), 63.3 (C-28), 77.0
(CH(Ph)2), 81.3 (C-3), 127.0, 127.1 (benzhydryl C-20, 60), 127.7
5.2.6.5.
3-O-(50-Benzhydryloxy-30,30-dimethylglutaryl)-28-O-
(300,300-dimethylglutaryl)betulin (26). Yield 100% (starting from
150 mg of 17 and 3,3-dimethylglutaric anhydride); white solid.
½
a 1D6 +7.3° (c 3.36, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.81 (3H,
ꢁ
s, CH3-24), 0.84 (3H, s, CH3-23, 25), 0.98 (3H, s, CH3-27), 1.04
(3H, s, CH3-26), 1.09 (6H, s, 3-O-dimethylglutaryl CH3), 1.16 (6H,
s, 28-O-dimethylglutaryl CH3), 1.70 (3H, s, CH3-30), 2.43–2.45
(1H, m, H-19), 2.36, 2.43 (each 1H, d, J = 14.4 Hz, 3-O-dimethyl-
glutaryl H2-20), 2.47, 2.48 (each 2H, s, 28-O-dimethylglutaryl H2-
200, 400), 2.55, 2.58 (each 1H, d, J = 14.4 Hz, 3-O-dimethylglutaryl
H2-40), 3.86, 4.29 (each 1H, d, J = 11.2 Hz, H2-28), 4.47 (1H, dd,
J = 4.8, 11.2 Hz, H-3), 4.60, 4.70 (each 1H, br s, H2-29), 6.89 (1H,
s, CH(Ph)2), 7.25–7.33 (10H, m, aromatic-H); 13C NMR (CDCl3,
100 MHz) d 14.7 (C-27), 16.0 (C-26), 16.1 (C-25), 16.6 (C-24),
18.1 (C-6), 19.1 (C-30), 20.8 (C-11), 23.8 (C-2), 25.2 (C-12), 27.0

6460
J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
(benzyhydryl C-40), 128.4 (benzhydryl C-30, 50), 140.3 (benzhydryl
C-10), 170.9 (3-O-dimethylsuccinyl 10-COOꢂ), 175.4 (3-O-dim-
ethylsuccinyl 40-COOꢂ, 28-O-dimethylsuccinyl 100-COOY), 177.0
(28-O-dimethylsuccinyl 100-COOꢂ). HRESIMS (positive) m/z
889.5590 [M+Na]+ (calcd for C55H78O8Na, 889.5594).
(each 3H, s, CH3-23, 24), 0.87 (3H, s, CH3-25), 0.96 (3H, s, CH3-
27), 1.05 (3H, s, CH3-26), 1.31 (6H, s, 28-O-dimethylsuccinyl
CH3), 2.00 (2H, quint, J = 7.2 Hz, 3-O-glutaryl H2-30), 2.36 (2H, t,
J = 7.2 Hz, 3-O-glutaryl H2-20), 2.50 (2H, t, J = 7.2 Hz, 3-O-glutaryl
H2-40), 2.65 (2H, s, 28-O-dimethylsuccinyl H2-300), 3.81, 4.32 (each
1H, d, J = 10.8 Hz, H2-28), 4.49 (1H, dd, J = 6.0, 10.0 Hz, H-3), 6.90
(1H, s, CH(Ph)2), 7.26–7.34 (10H, m, aromatic-H); 13C NMR (CDCl3,
100 MHz) d 14.7 (C-27), 14.9 (C-29), 16.1 (C-25, C-26), 16.6 (C-24),
18.2 (C-6), 20.4 (glutaryl C-30), 20.8 (C-11), 21.6 (C-21), 22.9 (C-30),
23.7 (C-2), 25.4, 25.5 (28-O-dimethylsuccinyl CH3), 26.8 (C-12),
26.9 (C-15), 28.0 (C-23), 29.4 (C-20), 29.9 (C-16), 33.6 (glutaryl
C-40), 33.7 (glutaryl C-20), 34.2 (C-7), 34.7 (C-22), 37.0 (C-10),
37.2 (C-13), 37.8 (C-4), 38.4 (C-1), 40.8 (28-O-dimethylsuccinyl
C-200), 40.9 (C-8), 42.9 (C-14), 43.9 (28-O-dimethylsuccinyl C-300),
44.6 (C-19), 46.7 (C-17), 48.2 (C-18), 50.0 (C-9), 55.4 (C-5), 63.4
(C-28), 77.0 (CH(Ph)2), 81.0 (C-3), 127.0 (benzhydryl C-20, 60),
127.9 (benzyhydryl C-40), 128.5 (benzhydryl C-30, 50), 140.2 (benz-
hydryl C-10), 171.9 (3-O-glutaryl 50-COOꢂ), 172.6 (3-O-glutaryl
10-COOꢂ), 175.8 (28-O-dimethylsuccinyl 100-COOY), 177.1 (28-O-
dimethylsuccinyl COOꢂ). HRESIMS (positive) m/z 875.5450
[M+Na]+ (calcd for C54H76O8Na, 875.5438).
5.2.6.7.
3-O-(40-Benzhydryloxy-30,30-dimethylsuccinyl)-28-O-
(300,300-dimethylglutaryl)dihydrobetulin (29). Yield 100% (start-
ing from 110 mg of 18 and 3,3-dimethylglutaric anhydride); white
solid. ½a 2D6
ꢁ
ꢂ10.2° (c 2.04, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.74
(3H, s, CH3-24), 0.78 (3H, s, CH3-23), 0.78, 0.85 (each 3H, d,
J = 8.0 Hz, CH3-29, 30), 0.80 (3H, s, CH3-25), 0.95 (3H, s, CH3-27),
1.04 (3H, s, CH3-26), 1.15 (6H, s, dimethylglutaryl CH3), 1.31,
1.32 (each 3H, s, dimethylsuccinyl CH3), 2.46, 2.48 (each 2H, s,
dimethylglutaryl H2-200, 400), 2.65, 2.71 (each 1H, d, J = 16.0 Hz,
3-O-dimethylsuccinyl H2-20), 3.83, 4.28 (each 1H, d, J = 10.8 Hz,
H2-28), 4.45 (1H, dd, J = 5.2, 11.2 Hz, H-3), 6.86 (1H, s, CH(Ph)2),
7.25–7.33 (10H, m, aromatic-H); 13C NMR (CDCl3, 100 MHz) d
14.6 (C-27), 14.9 (C-29), 16.0 (C-25, C-26), 16.5 (C-24), 18.1
(C-6), 20.7 (C-11), 21.6 (C-21), 22.9 (C-30), 23.5 (C-2), 25.2, 25.4
(3-O-dimethylsuccinyl CH3), 26.8 (C-12), 26.9 (C-15), 27.9 (C-23,
28-O-dimethylsuccinyl CH3), 29.4 (C-20), 29.9 (C-16), 32.7 (dim-
ethylglutaryl C-300), 34.1 (C-7), 34.7 (C-22), 37.0 (C-10), 37.2
(C-13), 37.6 (C-4), 38.4 (C-1), 40.7 (dimethylsuccinyl C-30), 40.9
(C-8), 42.8 (C-14), 44.5 (C-19, 3-O-dimethylsuccinyl C-20), 45.1
(dimethylglutaryl C-400), 45.4 (dimethylglutaryl C-200), 46.4 (C-17),
48.1 (C-18), 49.9 (C-9), 55.3 (C-5), 63.0 (C-28), 77.0 (CH(Ph)2),
81.3 (C-3), 127.0 (benzhydryl C-20, 60), 127.7 (benzyhydryl C-40),
128.4 (benzhydryl C-30, 50), 140.3 (benzhydryl C-10), 170.9
(3-O-dimethylsuccinyl 10-COOꢂ), 173.0 (28-O-dimethylglutaryl
100-COOꢂ), 175.4 (3-O-dimethylsuccinyl 40-COOꢂ), 176.0 (28-O-
dimethylglutaryl COOH). HRESIMS (positive) m/z 903.5748
[M+Na]+ (calcd for C56H80O8Na, 903.5751).
5.2.6.9. 3-O-(50-Benzhydryloxyglutaryl)-28-O-glutaryldihydro-
betulin (32). Yield 94.9% (starting from 89 mg of 20 and glutaric
anhydride); colorless oil. ½a D27
ꢁ
ꢂ1.2° (c 1.04, CHCl3). 1H NMR (CDCl3,
400 MHz) d 0.78, 0.84 (each 3H, d, J = 8.0 Hz, CH3-29, 30), 0.83 (3H,
s, CH3-24), 0.84 (3H, s, CH3-24), 0.87 (3H, s, CH3-25), 0.96 (3H, s,
CH3-27), 1.06 (3H, s, CH3-26), 1.98 (2H, quint, J = 7.2 Hz, 28-O-glut-
aryl H2-300), 2.00 (2H, quint, J = 7.2 Hz, 3-O-glutaryl H2-30), 2.36
(2H, t, J = 7.2 Hz, 3-O-glutaryl H2-20), 2.43 (2H, t, J = 7.2 Hz, 28-O-
glutaryl H2-200), 2.44 (2H, t, J = 7.2 Hz, 28-O-glutaryl H2-400), 2.50
(2H, t, J = 7.2 Hz, 3-O-glutaryl H2-40), 3.84, 4.30 (each 1H, d,
J = 10.8 Hz, H2-28), 4.49 (1H, dd, J = 6.0, 10.4 Hz, H-3), 6.90 (1H, s,
CH(Ph)2), 7.26–7.33 (10H, m, aromatic-H); 13C NMR (CDCl3,
100 MHz) d 14.6 (C-27), 14.9 (C-29), 16.1 (C-25, C-26), 16.6 (C-
24), 18.2 (C-6), 20.0 (28-O-glutaryl C-300), 20.4 (3-O-glutaryl C-30),
20.8 (C-11), 21.6 (C-21), 22.9 (C-30), 23.7 (C-2), 26.8 (C-12), 27.0
(C-15), 28.0 (C-23), 29.4 (C-20), 29.9 (C-16), 32.8 (28-O-glutaryl
C-400), 33.3 (28-O-glutaryl C-200), 33.6 (3-O-glutaryl C-40), 33.7 (3-
O-glutaryl C-20), 34.2 (C-7), 34.7 (C-22), 37.1 (C-10), 37.2 (C-13),
37.8 (C-4), 38.4 (C-1), 41.0 (C-8), 42.9 (C-14), 44.6 (C-19), 46.6
(C-17), 48.2 (C-18), 50.0 (C-9), 55.4 (C-5), 62.9 (C-28), 77.0
(CH(Ph)2), 81.0 (C-3), 127.1 (benzhydryl C-20, 60), 127.9 (benzyhyd-
ryl C-40), 128.5 (benzhydryl C-30, 50), 140.2 (benzhydryl C-10), 171.9
(3-O-glutaryl 50-COOꢂ), 172.6 (3-O-glutaryl 10-COOꢂ), 173.3
(28-O-glutaryl 100-COOꢂ), 177.3 (28-O-glutaryl COOY). HRESIMS
(positive) m/z 861.5292 [M+Na]+ (calcd for C53H74O8Na, 861.5281).
5.2.6.8. 3-O-(50-Benzhydryloxyglutaryl)-28-O-(300,300-dimethyl-
succinyl)dihydrobetulin (30) and 3-O-(50-benzhydryloxygluta-
ryl)-28-O-(200,200-dimethylsuccinyl)dihydrobetulin (31). Total
yield 98.9% (starting from 225 mg of 20 and 2,2-dimethylsuccinic
acid); used for next reaction without purification. For spectroscopic
analysis, a small amount of sample was further purified by prepara-
tive HPLC (pNAP, CH3OH/H2O/CH3COOH = 95:4:1); colorless oil.
Compound 30: ½a 2D7
ꢁ
ꢂ7.1° (c 3.50, CHCl3). 1H NMR (CDCl3,
400 MHz) d 0.78, 0.85 (each 3H, d, J = 8.0 Hz, CH3-29, 30), 0.84
(each 3H, s, CH3-23, 24), 0.87 (3H, s, CH3-25), 0.96 (3H, s, CH3-
27), 1.05 (3H, s, CH3-26), 1.32 (6H, s, 28-O-dimethylsuccinyl
CH3), 2.00 (2H, quint, J = 7.2 Hz, 3-O-glutaryl H2-30), 2.36 (2H, t,
J = 7.2 Hz, 3-O-glutaryl H2-20), 2.51 (2H, t, J = 7.2 Hz, 3-O-glutaryl
H2-40), 2.65 (2H, s, 28-O-dimethylsuccinyl H2-200), 3.85, 4.28 (each
1H, d, J = 10.8 Hz, H2-28), 4.49 (1H, dd, J = 6.0, 10.0 Hz, H-3), 6.90
(1H, s, CH(Ph)2), 7.26–7.34 (10H, m, aromatic-H); 13C NMR (CDCl3,
100 MHz) d 14.6 (C-27), 14.9 (C-29), 16.1 (C-25, C-26), 16.5 (C-24),
18.2 (C-6), 20.3 (glutaryl C-30), 20.8 (C-11), 21.6 (C-21), 22.9 (C-30),
23.7 (C-2), 25.3 (28-O-dimethylsuccinyl CH3), 26.8 (C-12), 26.9 (C-
15), 28.0 (C-23), 29.4 (C-20), 29.8 (C-16), 33.6 (glutaryl C-40), 33.7
(glutaryl C-20), 34.2 (C-7), 34.7 (C-22), 37.0 (C-10), 37.2 (C-13), 37.8
(C-4), 38.4 (C-1), 40.5 (28-O-dimethylsuccinyl C-300), 40.9 (C-8),
42.9 (C-14), 44.4 (28-O-dimethylsuccinyl C-200), 44.5 (C-19), 46.4
(C-17), 48.2 (C-18), 50.0 (C-9), 55.3 (C-5), 63.1 (C-28), 76.9
(CH(Ph)2), 81.0 (C-3), 127.0 (benzhydryl C-20, 60), 127.9 (benzyhyd-
ryl C-40), 128.5 (benzhydryl C-30, 50), 140.2 (benzhydryl C-10), 171.5
(28-O-dimethylsuccinyl 100-COOꢂ), 171.9 (3-O-glutaryl 50-COOꢂ),
172.6 (3-O-glutaryl 10-COOꢂ), 182.6 (28-O-dimethylsuccinyl
COOH). HRESIMS (positive) m/z 875.5408 [M+Na]+ (calcd for
5.2.6.10. 3-O-(50-Benzhydryloxyglutaryl)-28-O-(300,300-dimethyl-
glutaryl)dihydrobetulin (33). Yield 91.6% (starting from 82 mg
of 20 and 3,3-dimethylglutaric anhydride); colorless oil. ½a D27
ꢁ
ꢂ5.4° (c 1.43, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.78, 0.84 (each
3H, d, J = 8.0 Hz, CH3-29, 30), 0.83 (3H, s, CH3-24), 0.84 (3H, s, CH3-
24), 0.87 (3H, s, CH3-25), 0.97 (3H, s, CH3-27), 1.06 (3H, s, CH3-26),
1.16 (6H, s, 28-O-dimethylglutaryl CH3), 2.00 (2H, quint, J = 7.2 Hz,
3-O-glutaryl H2-30), 2.36 (2H, t, J = 7.2 Hz, 3-O-glutaryl H2-20), 2.46
(2H, s, 28-O-dimthylglutaryl H2-200), 2.49 (2H, s, 28-O-glutaryl H2-
400), 2.50 (2H, t, J = 7.2 Hz, 3-O-glutaryl H2-40), 3.84, 4.30 (each 1H,
d, J = 10.8 Hz, H2-28), 4.49 (1H, dd, J = 5.6, 10.8 Hz, H-3), 6.90 (1H, s,
CH(Ph)2), 7.26–7.33 (10H, m, aromatic-H); 13C NMR (CDCl3,
100 MHz) d 14.7 (C-27), 14.9 (C-29), 16.1 (C-25, C-26), 16.6 (C-
24), 18.2 (C-6), 20.4 (3-O-glutaryl C-30), 20.8 (C-11), 21.6 (C-21),
22.9 (C-30), 23.7 (C-2), 26.8 (C-12), 26.9 (C-15), 27.9 (28-O-dim-
ethylglutaryl CH3), 28.0 (C-23), 29.4 (C-20), 29.9 (C-16), 32.7
(28-O-dimethylglutaryl C-300), 33.6 (3-O-glutaryl C-40), 33.7 (3-O-
glutaryl C-20), 34.2 (C-7), 34.7 (C-22), 37.1 (C-10), 37.2 (C-13),
C
54H76O8Na, 875.5438).
Compound 31: ½a 2D7
ꢁ
ꢂ7.4° (c 1.26, CHCl3). 1H NMR (CDCl3,
400 MHz) d 0.79, 0.85 (each 3H, d, J = 8.0 Hz, CH3-29, 30), 0.84

J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
6461
37.8 (C-4), 38.4 (C-1), 41.0 (C-8), 42.9 (C-14), 44.6 (C-19), 45.1 (28-
O-dimethylglutaryl C-400), 45.4 (28-O-dimethylglutaryl C-200), 46.4
(C-17), 48.2 (C-18), 50.0 (C-9), 55.4 (C-5), 63.1 (C-28), 77.0
(CH(Ph)2), 81.0 (C-3), 127.1 (benzhydryl C-20, 60), 127.9 (benzyhyd-
ryl C-40), 128.5 (benzhydryl C-30, 50), 140.2 (benzhydryl C-10), 171.9
(3-O-glutaryl 50-COOꢂ), 172.6 (3-O-glutaryl 10-COOꢂ), 173.0
(28-O-dimethylglutaryl 100-COOꢂ), 175.6 (28-O-dimethylglutaryl
COOY). HRESIMS (positive) m/z 889.5605 [M+Na]+ (calcd for
16.0 (C-26), 16.1 (C-25), 16.5 (C-24), 18.1 (C-6), 19.1 (C-30), 20.7
(C-11), 23.5 (C-2), 25.1 (C-12), 25.2, 25.4 (3-O-dimethylsuccinyl
CH3), 27.0 (C-15), 27.9 (C-23), 29.0 (28-O-succinyl C-200, 300), 29.5
(C-21), 29.7 (C-16), 34.1 (C-7), 34.5 (C-22), 37.0 (C-10), 37.6
(AZT-C-20), 37.6 (C-13), 37.7 (C-4), 38.3 (C-1), 40.7 (dimethylsucci-
nyl C-30), 40.8 (C-8), 42.7 (C-14), 44.5 (3-O-dimethylsuccinyl C-20),
46.4 (C-17), 47.7 (C-19), 48.7 (C-18), 50.2 (C-9), 55.3 (C-5), 60.1
(AZT-C-30), 63.1 (AZT-C-50), 63.3 (C-28), 77.0 (CH(Ph)2), 81.3 (C-
3), 81.9 (AZT-C-40), 85.1 (AZT-C-10), 110.0 (C-29), 111.3 (AZT-C-
5), 127.0, 127.1 (benzhydryl C-20, 60), 127.7 (benzyhydryl C-40),
128.4 (benzhydryl C-30, 50), 135.3 (AZT-C-6), 140.3 (benzhydryl C-
10), 149.8 (AZT-C-2), 150.0 (C-20), 163.1 (AZT-C-4), 170.9 (3-O-
dimethylsuccinyl 10-COOꢂ), 172.0 (28-O-succinyl 400-COO-AZT),
172.6 (28-O-succinyl 100-COOꢂ), 175.4 (3-O-dimethylsuccinyl 40-
COOꢂ). HRESIMS (positive) m/z 1108.6014 [M+Na]+ (calcd for
C55H78O8Na, 889.5594).
5.2.6.11.
3-O-(50-Benzhydryloxy-30,30-dimethylglutaryl)-28-O-
(300,300-dimethylglutaryl)dihydrobetulin (34). Yield 97% (starting
from 137 mg of 19 and 3,3-dimethylglutaric anhydride); white so-
lid. ½a 2D8
ꢁ
ꢂ9.3° (c 2.94, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.78,
0.83 (each 3H, d, J = 8.0 Hz, CH3-29, 30), 0.81 (3H, s, CH3-24),
0.84 (3H, s, CH3-24), 0.85 (3H, s, CH3-25), 0.96 (3H, s, CH3-27),
1.05 (3H, s, CH3-26), 1.09 (6H, s, 3-O-dimethylglutaryl CH3), 1.15
(6H, s, 28-O-dimethylglutaryl CH3), 2.36, 2.43 (each 1H, d,
J = 14.4 Hz, 3-O-dimethylglutaryl H2-20), 2.46 (2H, s, 28-O-dimthyl-
glutaryl H2-200), 2.48 (2H, s, 28-O-dimethylglutaryl H2-400), 2.55,
2.58 (each 1H, d, J = 14.4 Hz, 3-O-dimethylglutaryl H2-40), 3.83,
4.29 (each 1H, d, J = 11.2 Hz, H2-28), 4.47 (1H, dd, J = 4.8, 10.8 Hz,
H-3), 6.89 (1H, s, CH(Ph)2), 7.25–7.34 (10H, m, aromatic-H); 13C
NMR (CDCl3, 100 MHz) d 14.6 (C-27), 14.9 (C-29), 16.0 (C-25, C-
26), 16.6 (C-24), 18.1 (C-6), 20.8 (C-11), 21.6 (C-21), 22.9 (C-30),
23.7 (C-2), 26.8 (C-12, C-15), 27.6, 27.7 (3-O-dimethylglutaryl
CH3), 27.9 (28-O-dimethylglutaryl CH3), 28.0 (C-23), 29.4 (C-20),
29.9 (C-16), 32.6 (28-O-dimethylglutaryl C-300), 32.8 (3-O-dimeth-
ylglutaryl C-30), 34.2 (C-7), 34.7 (C-22), 37.0 (C-10), 37.2 (C-13),
37.6 (C-4), 38.4 (C-1), 41.0 (C-8), 42.8 (C-14), 44.5 (C-19), 45.1
(28-O-dimethylglutaryl C-400), 45.3 (28-O-dimethylglutaryl C-200),
45.4 (3-O-dimethylglutaryl C-40), 45.8 (3-O-dimethylglutaryl C-
20), 46.4 (C-17), 48.1 (C-18), 49.9 (C-9), 55.3 (C-5), 63.0 (C-28),
76.7 (CH(Ph)2), 80.9 (C-3), 127.1 (benzhydryl C-20, 60), 127.8
(benzyhydryl C-40), 128.4 (benzhydryl C-30, 50), 140.2, 140.3
(benzhydryl C-10), 170.8 (3-O-dimethylglutaryl 50-COOꢂ), 171.7
(3-O-dimethylglutaryl 10-COOꢂ), 172.9 (28-O-dimethylglutaryl
100-COOꢂ), 176.4 (28-O-dimethylglutaryl COOY). HRESIMS (posi-
tive) m/z 917.5843 [M+Na]+ (calcd for C57H82O8Na, 917.5907).
C63H83N5O11Na, 1108.5987).
5.2.7.2. 1-(30-Azido-30-deoxythymidine-50-yl)-4-[3-O-(40-benz-
hydryloxy-30,30-dimethylsuccinyl)-lup-20(29)en-28-yl] 2,2-dim-
ethylsuccinate (36) and 4-(30-Azido-30-deoxythymidine-50-yl)-1-
[3-O-(40-benzhydryloxy-30,30-dimethylsuccinyl)-lup-20(29)en-
28-yl] 2,2-dimethylsuccinate (37). Total yield 49.3% (starting
from 296 mg of mixture of 22 and 23); used for next reaction with-
out separation. A small amount of sample mixture was separated
by HPLC (cholester, CH3OH/H2O = 97:3) for spectroscopic analyses.
Compound 36: ½a 2D7
ꢁ
+7.2° (c 1.77, CHCl3). 1H NMR (CDCl3,
400 MHz) d 0.74 (3H, s, CH3-24), 0.78 (each 3H, s, CH3-23, 25),
0.96 (3H, s, CH3-27), 1.02 (3H, s, CH3-26), 1.30, 1.31 (each 3H, s,
28-O-dimethylsuccinyl CH3), 1.31, 1.32 (each 3H, s, 3-O-dim-
ethylsuccinyl CH3), 1.68 (3H, s, CH3-30), 1.94 (3H, br s, AZT-CH3),
2.26–2.33, 2.44–2.50 (each 1H, m, AZT-H2-20), 2.37–2.44 (1H, m,
H-19), 2.65, 2.69 (each 1H, d, J = 16.0 Hz, 3-O-dimethylsuccinyl
H2-20), 2.61, 2.75 (each 1H, d, J = 16.4 Hz, 28-O-dimethylsuccinyl
H2-200), 3.84, 4.29 (each 1H, d, J = 10.8 Hz, H2-28), 4.09 (1H, q-like,
AZT-H-40), 4.26–4.30 (1H, m, AZT-H-30), 4.30, 4.52 (each 1H, dd,
J = 4.0, 12.0 Hz, AZT-H2-50), 4.43 (1H, dd, J = 5.2, 10.8 Hz, H-3),
4.60, 4.69 (each 1H, br s, H2-29), 6.18 (1H, t-like, J = 6.4 Hz, AZT-
H-10), 6.86 (1H, s, CH(Ph)2), 7.24–7.33 (11H, m, aromatic-H and
AZT-H-6), 8.56 (1H, br s, AZT-NH); 13C NMR (CDCl3, 100 MHz) d
12.5 AZT-CH3, 14.7 (C-27), 15.9 (C-26), 16.1 (C-25), 16.5 (C-24),
18.1 (C-6), 19.1 (C-30), 20.7 (C-11), 23.5 (C-2), 25.2 (C-12), 25.2,
25.4 (3-O-dimethylsuccinyl CH3), 25.1, 25.9 (28-O-dimethylsucci-
nyl CH3), 27.0 (C-15), 27.9 (C-23), 29.5 (C-21), 29.7 (C-16), 34.1
(C-7), 34.5 (C-22), 37.0 (C-10), 37.4 (AZT-C-20), 37.6 (C-4, 13),
38.3 (C-1), 40.7 (dimethylsuccinyl C-30, C-300), 40.8 (C-8), 42.7 (C-
14), 44.5 (3-O-dimethylsuccinyl C-20), 44.6 (28-O-dimethylsuccinyl
C-200), 46.3 (C-17), 47.7 (C-19), 48.7 (C-18), 50.2 (C-9), 55.3 (C-5),
60.4 (AZT-C-30), 63.1 (C-28), 63.0 (AZT-C-50), 77.0 (CH(Ph)2), 81.2
(C-3), 81.9 (AZT-C-40), 84.8 (AZT-C-10), 110.0 (C-29), 111.4 (AZT-
C-5), 127.0, 127.1 (benzhydryl C-20, 60), 127.7 (benzyhydryl C-40),
128.4 (benzhydryl C-30, 50), 135.0 (AZT-C-6), 140.4 (benzhydryl C-
10), 149.9 (C-20 and AZT-C-2), 163.3 (AZT-C-4), 170.8 (3-O-dim-
ethylsuccinyl 10-COOꢂ), 171.7 (28-O-dimethylsuccinyl 100-COOꢂ),
175.4 (3-O-dimethylsuccinyl 40-COOꢂ), 176.4 (28-O-dim-
ethylsuccinyl 400-COO-AZT). HRESIMS (positive) m/z 1136.6310
[M+Na]+ (calcd for C65H87N5O11Na, 1136.6300).
5.2.7. General procedure for coupling of 3,28-di-O-acylbetulin
derivatives with AZT (35–48)
To a solution of 3,28-di-O-acylbetulin derivatives (1 equiv) in
CH2Cl2 (4–10 mL) was added DMAP (2 equiv), DCC (2 equiv) and
30-azido-30-dexoythymidine (AZT, 2 equiv). The reaction mixture
was kept stirring at room temperature for overnight. After filtra-
tion, the filtrate was concentrated and the residue was chromato-
graphed over silica gel column (hexane/EtOAc = 2:1) or purified by
HPLC.
5.2.7.1. 30-Azido-30-deoxythymidine 50-yl 3-O-(40-benzhydryloxy-
30,30-dimethylsuccinyl)-lup-20(29)en-28-yl succinate (35). Yield
41.7% (starting from 80 mg of 21); white solid. ½a D18
ꢁ
+6.2° (c 0.53,
CHCl3). 1H NMR (CDCl3, 500 MHz) d 0.72 (3H, s, CH3-24), 0.76
(3H, s, CH3-23), 0.77 (3H, s, CH3-25), 0.95 (3H, s, CH3-27), 1.01
(3H, s, CH3-26), 1.29, 1.30 (each 3H, s, 3-O-dimethylsuccinyl
CH3), 1.68 (3H, s, CH3-30), 1.95 (3H, br s, AZT-CH3), 2.33–2.50
(3H, m, AZT-H2-20 and H-19), 2.62–2.79 (4H, m, 28-O-succinyl
H2-200, 300), 2.64, 2.68 (each 1H, d, J = 16.0 Hz, 3-O-dimethylsuccinyl
H2-20), 3.86, 4.30 (each 1H, d, J = 11.2 Hz, H2-28), 4.05 (1H, q-like,
AZT-H-40), 4.21–4.24 (1H, m, AZT-H-30), 4.30 (1H, dd, J = 3.4,
12.0 Hz, AZT-H2-5a0), 4.52 (1H, dd, J = 4.0, 12.0 Hz, AZT-H2-5b0),
4.42 (1H, dd, J = 5.2, 11.2 Hz, H-3), 4.59, 4.68 (each 1H, br s, H2-
29), 6.15 (1H, t-like, J = 6.3 Hz, AZT-H-10), 6.84 (1H, s, CH(Ph)2),
7.24–7.32 (11H, m, aromatic-H and AZT-H-6), 8.13 (1H, br s,
AZT-NH); 13C NMR (CDCl3, 125 MHz) d 12.6 AZT-CH3, 14.7 (C-27),
Compound 37: ½a 2D7
ꢁ
+11.6° (c 3.64, CHCl3). 1H NMR (CDCl3,
400 MHz) d 0.73 (3H, s, CH3-24), 0.77 (3H, s, CH3-23), 0.78 (3H, s,
C-25), 0.96 (3H, s, CH3-27), 1.02 (3H, s, CH3-26), 1.30, 1.32 (each
3H, s, 3-O-dimethylsuccinyl CH3), 1.31, 1.34 (each 3H, s, 28-O-dim-
ethylsuccinyl CH3), 1.69 (3H, s, CH3-30), 1.95 (3H, br s, AZT-CH3),
2.33–2.50 (3H, m, AZT-H2-20 and H-19), 2.62, 2.66 (each 1H, d,
J = 16.4 Hz, 28-O-dimethylsuccinyl H2-300), 2.64, 2.69 (each 1H, d,
J = 16.0 Hz, 3-O-dimethylsuccinyl H2-20), 3.85, 4.29 (each 1H, d,
J = 10.8 Hz, H2-28), 4.04 (1H, q-like, AZT-H-40), 4.18–4.22 (1H, m,
AZT-H-30), 4.26 (1H, dd, J = 4.0, 12.0 Hz, AZT-H-5a0), 4.42 (1H, dd,

6462
J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
J = 4.4, 12.0 Hz, AZT-H-5b0), 4.43 (1H, dd, J = 4.8, 10.4 Hz, H-3), 4.60,
4.69 (each 1H, br s, H2-29), 6.15 (1H, t-like, J = 6.4 Hz, AZT-H-10),
6.85 (1H, s, CH(Ph)2), 7.24–7.32 (11H, m, aromatic-H and AZT-H-
6), 8.94 (1H, br s, AZT-NH); 13C NMR (CDCl3, 100 MHz) d 12.6
AZT-CH3, 14.7 (C-27), 15.9 (C-26), 16.0 (C-25), 16.5 (C-24), 18.1
(C-6), 19.1 (C-30), 20.7 (C-11), 23.5 (C-2), 25.2 (C-12), 25.2, 25.5
(3-O-dimethylsuccinyl CH3), 25.4, 25.8 (28-O-dimethylsuccinyl
CH3), 27.0 (C-15), 27.9 (C-23), 29.5 (C-21), 29.8 (C-16), 34.1 (C-7),
34.5 (C-22), 37.0 (C-10), 37.5 (AZT-C-20), 37.6 (C-4, 13), 38.3
(C-1), 40.7 (3-O-dimethylsuccinyl C-30), 40.8 (C-8), 41.0 (28-O-dim-
ethylsuccinyl C-200), 42.7 (C-14), 43.8 (28-O-dimethylsuccinyl
C-300), 44.5 (3-O-dimethylsuccinyl C-20), 46.5 (C-17), 47.7 (C-19),
48.8 (C-18), 50.2 (C-9), 55.3 (C-5), 60.3 (AZT-C-30), 62.9 (AZT-C-
50), 63.2 (C-28), 77.0 (CH(Ph)2), 81.2 (C-3), 81.7 (AZT-C-40), 85.2
(AZT-C-10), 109.9 (C-29), 111.3 (AZT-C-5), 127.0, 127.1 (benzhydryl
C-20, 60), 127.7 (benzyhydryl C-40), 128.4 (benzhydryl C-30, 50),
135.3 (AZT-C-6), 140.3 (benzhydryl C-10), 150.0 (C-20 and AZT-C-
2), 163.5 (AZT-C-4), 170.7 (28-O-dimethylsuccinyl 400-COO-AZT),
170.8 (3-O-dimethylsuccinyl 10-COOꢂ), 175.4 (3-O-dimethylsucci-
nyl 40-COOꢂ), 176.9 (28-O-dimethylsuccinyl 100-COOꢂ). HRESIMS
(positive) m/z 1136.6299 [M+Na]+ (calcd for C65H87N5O11Na,
1136.6300).
1.69 (3H, s, CH3-30), 1.95 (3H, br s, AZT-CH3), 2.30–2.35, 2.40–2.51
(2H, m, AZT-H2-20), 2.35–2.42 (1H, m, H-19), 2.43, 2.50 (each 1H, d,
J = 16.0 Hz, 28-O-dimethylglutaryl H2-200), 2.49, 2.56 (each 1H, d,
J = 16.0 Hz, 28-O-dimethylglutaryl H2-400), 2.65, 2.69 (each 1H, d,
J = 16.0 Hz, 3-O-dimethylsuccinyl H2-20), 3.82, 4.26 (each 1H, d,
J = 10.8 Hz, H2-28), 4.07 (1H, q-like, AZT-H-40), 4.20–4.25 (1H, m,
AZT-H-30), 4.27 (1H, dd, J = 3.6, 12.0 Hz, AZT-H-5a0), 4.44 (1H, dd,
J = 4.4, 12.0 Hz, AZT-H-5b0), 4.44 (1H, dd, J = 4.4, 12.0 Hz, H-3),
4.60, 4.69 (each 1H, br s, H2-29), 6.15 (1H, t-like, J = 6.4 Hz, AZT-
H-10), 6.86 (1H, s, CH(Ph)2), 7.24–7.33 (11H, m, aromatic-H and
AZT-H-6), 8.78 (1H, br s, AZT-NH); 13C NMR (CDCl3, 100 MHz) d
12.6 AZT-CH3, 14.7 (C-27), 16.0 (C-26), 16.1 (C-25), 16.5 (C-24),
18.1 (C-6), 19.1 (C-30), 20.8 (C-11), 23.5 (C-2), 25.2 (C-12), 25.2,
25.4 (3-O-dimethylsuccinyl CH3), 27.0 (C-15), 27.9 (28-O-dimeth-
ylglutaryl CH3), 28.0 (C-23), 29.5 (C-21), 29.8 (C-16), 32.6 (28-O-
dimethylglutaryl C-300), 34.1 (C-7), 34.6 (C-22), 37.0 (C-10), 37.5
(AZT-C-20), 37.6 (C-4, 13), 38.3 (C-1), 40.7 (3-O-dimethylsuccinyl
C-30), 40.8 (C-8), 42.6 (C-14), 44.5 (3-O-dimethylsuccinyl C-20, 28-
O-dimethylglutaryl C-400), 45.0 (28-O-dimethylglutaryl C-200), 46.2
(C-17), 47.7 (C-19), 48.7 (C-18), 50.2 (C-9), 55.3 (C-5), 60.4 (AZT-
C-30), 62.6 (C-28), 62.7 (AZT-C-50), 77.0 (CH(Ph)2), 81.2 (C-3), 81.8
(AZT-C-40), 85.2 (AZT-C-10), 109.9 (C-29), 111.3 (AZT-C-5), 127.0,
127.1 (benzhydryl C-20, 60), 127.7 (benzyhydryl C-40), 128.4 (benz-
hydryl C-30, 50), 135.2 (AZT-C-6), 140.3 (benzhydryl C-10), 150.0
(C-20 and AZT-C-2), 163.4 (AZT-C-4), 170.8 (3-O-dimethylsuccinyl
10-COOꢂ), 171.1 (28-O-dimethylglutaryl 500-COO-AZT), 172.3
(28-O-dimethylglutaryl 100-COOꢂ), 175.4 (3-O-dimethylsuccinyl
40-COOꢂ). HRESIMS (positive) m/z 1150.6467 [M+Na]+ (calcd for
5.2.7.3. 30-Azido-30-deoxythymidine-50-yl 3-O-(40-benzhydryl-
oxy-30,30-dimethylsuccinyl)-lup-20(29)en-28-yl glutarate (38).
Yield 47.4% (starting from 180 mg of 24); white solid. ½a D15
ꢁ
+24.2°
(c 0.5, CHCl3). 1H NMR (CDCl3, 500 MHz) d 0.73 (3H, s, CH3-24),
0.77 (3H, s, CH3-23), 0.78 (3H, s, CH3-25), 0.96 (3H, s, CH3-27),
1.01 (3H, s, CH3-26), 1.30, 1.31 (each 3H, s, 3-O-dimethylsuccinyl
CH3), 1.68 (3H, s, CH3-30), 1.94 (3H, s, AZT-CH3), 1.99 (2H, quint,
J = 7.0 Hz, 28-O-glutaryl H2-300), 2.42 (2H, t, J = 7.0 Hz, 28-O-gluta-
ryl H2-200), 2.47 (2H, t, J = 7.2 Hz, 28-O-glutaryl H2-400), 2.33–2.50
(3H, m, AZT-H2-20 and H-19), 2.64, 2.69 (each 1H, d, J = 16.0 Hz,
3-O-dimethylsuccinyl H2-20), 3.85, 4.28 (1H, d, J = 11.2 Hz, H2-
28a), 4.06 (1H, m, AZT-H-40), 4.23 (1H, m, AZT-H-30), 4.32 (1H,
dd, J = 3.5, 12.2 Hz, AZT-H-5a0), 4.39 (1H, dd, J = 4.8, 12.2 Hz, AZT-
H-5b0), 4.43 (1H, dd, J = 4.8, 11.2 Hz, H-3), 4.59, 4.69 (each 1H, br
s, H2-29), 6.10 (1H, t-like, J = 5.5 Hz, AZT-H-10), 6.85 (1H, s,
CH(Ph)2), 7.21–7.32 (11H, m, aromatic-H and AZT-H-6), 9.37 (1H,
br s, AZT-NH); 13C NMR (CDCl3, 125 MHz) 13C NMR (CDCl3,
125 MHz) d 12.6 AZT-CH3, 14.7 (C-27), 15.9 (C-26), 16.0 (C-25),
16.4 (C-24), 18.0 (C-6), 19.0 (C-30), 19.9 (28-O-glutaryl
C-300), 20.7 (C-11), 23.5 (C-2), 25.0 (C-12), 25.1, 25.3 (3-O-dim-
ethylsuccinyl CH3), 26.9 (C-15), 27.8 (C-23), 29.4 (C-21), 29.7
(C-16), 33.0 (28-O-glutaryl C-400), 33.1 (28-O-glutaryl C-200), 34.0
(C-7), 34.4 (C-22), 36.9 (C-10), 37.4 (AZT-C-20), 37.5 (C-13), 37.6
(C-4), 38.3 (C-1), 40.6 (dimethylsuccinyl C-30), 40.8 (C-8), 42.6
(C-14), 44.4 (3-O-dimethylsuccinyl C-20), 46.3 (C-17), 47.6 (C-19),
48.7 (C-18), 50.1 (C-9), 55.2 (C-5), 60.5 (AZT-C-30), 62.8 (C-28),
63.3 (AZT-C-50), 76.9 (CH(Ph)2), 81.2 (C-3), 81.6 (AZT-C-40), 85.6
(AZT-C-10), 109.9 (C-29), 111.2 (AZT-C-5), 126.9, 127.0 (benzhydryl
C-20, 60), 127.7 (benzyhydryl C-40), 128.3 (benzhydryl C-30, 50),
135.4 (AZT-C-6), 140.2, 140.3 (benzhydryl C-10), 149.9 (C-20),
150.0 (AZT-C-2), 163.4 (AZT-C-4), 170.8 (3-O-dimethylsuccinyl
10-COOꢂ), 172.3 (28-O-glutaryl 500-COO-AZT), 173.1 (28-O-glutaryl
100-COOꢂ), 175.3 (3-O-dimethylsuccinyl 40-COOꢂ). HRESIMS (posi-
tive) m/z 1122.6141 [M+Na]+ (calcd for C64H85N5O11Na,
1122.6143).
C66H89N5O11Na, 1150.6456).
5.2.7.5. 300-Azido-30-deoxythymidine-50-yl 3-O-(40-benzhydryl-
oxy-30,30-dimethylglutaryl)-lup-20(29)en-28-yl 3,3-dimethylglu-
tarate (40). Yield 58.8% (starting from 143 mg of 26); white solid.
½
a 2D6 +13.5° (c 4.13, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.80 (3H,
ꢁ
s, CH3-24), 0.82 (6H, s, CH3-23, 25), 0.97 (3H, s, CH3-27), 1.02 (3H,
s, CH3-26), 1.08 (6H, s, 3-O-dimethylglutaryl CH3), 1.12, 1.15 (each
3H, s, 28-O-dimethylglutaryl CH3), 1.68 (3H, s, CH3-30), 1.95
3H, br s, AZT-CH3), 2.32–2.50 (3H, m, AZT-H2-20 and H-19), 2.34,
2.41 (each 1H, d, J = 14.4 Hz, 3-O-dimethylglutaryl H2-20), 2.44,
2.50 (each 1H, d, J = 14.4 Hz, 28-O-dimethylglutaryl H2-200), 2.49,
2.56 (each 1H, d, J = 14.4 Hz, 28-O-dimethylglutaryl H2-300), 2.54,
2.57 (each 1H, d, J = 14.4 Hz, 3-O-dimethylglutaryl H2-40), 3.82,
4.26 (each 1H, d, J = 11.2 Hz, H2-28), 4.06 (1H, q-like, AZT-H-40),
4.21–4.28 (1H, m, AZT-H-30), 4.26 (1H, dd, J = 3.6, 12.0 Hz, AZT-
H-5a0), 4.43 (1H, dd, J = 4.4, 12.0 Hz, AZT-H-5b0), 4.46 (1H, dd,
J = 4.8, 11.2 Hz, H-3), 4.59, 4.69 (each 1H, br s, H2-29), 6.15 (1H,
t-like, J = 6.4 Hz, AZT-H-10), 6.88 (1H, s, CH(Ph)2), 7.24–7.33
(11H, m, aromatic-H and AZT-H-6), 9.07 (1H, br s, AZT-NH); 13C
NMR (CDCl3, 100 MHz) d 12.5 AZT-CH3, 14.7 (C-27), 15.9 (C-26),
16.1 (C-25), 16.6 (C-24), 18.1 (C-6), 19.1 (C-30), 20.7 (C-11),
23.7 (C-2), 25.1 (C-12), 26.9 (C-15), 27.6, (3-O-dimethylglutaryl
CH3), 27.8, 27.9 (28-O-dimethylglutaryl CH3), 27.9 (C-23), 29.5
(C-21), 29.8 (C-16), 32.5 (28-O-dimethylglutaryl C-300), 32.8 (3-
O-dimethylglutaryl C-30), 34.1 (C-7), 34.6 (C-22), 37.0 (C-10),
37.5 (AZT-C-20, C-13), 37.6 (C-4), 38.3 (C-1), 40.8 (C-8), 42.6
(C-14), 44.5 (28-O-dimethylglutaryl C-400), 45.0 (28-O-dim-
ethylsuccinyl C-200), 45.4 (3-O-dimethylglutaryl C-40), 45.7 (3-O-
dimethylglurayl C-20), 46.2 (C-17), 47.6 (C-19), 48.7 (C-18), 50.2
(C-9), 55.3 (C-5), 60.4 (AZT-C-30), 62.6 (C-28), 62.7 (AZT-C-50),
76.6 (CH(Ph)2), 80.8 (C-3), 81.7 (AZT-C-40), 85.2 (AZT-C-10), 109.9
(C-29), 111.3 (AZT-C-5), 127.1 (benzhydryl C-20, 60), 127.7 (ben-
zyhydryl C-40), 128.4 (benzhydryl C-30, 50), 135.2 (AZT-C-6),
140.2 (benzhydryl C-10), 149.9 (C-20), 150.0 (AZT-C-2), 163.5
(AZT-C-4), 170.8 (3-O-dimethylglutaryl 50-COOꢂ), 171.1 (28-O-
dimethylglutaryl 500-COO-AZT), 171.6 (3-O-dimethylglutaryl
10-COOꢂ), 172.2 (28-O-dimethylglutaryl 100-COOꢂ). HRESIMS
5.2.7.4. 30-Azido-30-deoxythymidine-50-yl 3-O-(40-benzhydryl-
oxy-30,30-dimethylsuccinyl)-lup-20(29)en-28-yl 3,3-dimethyl-
glutarate (39). Yield 74.2% (starting from 120 mg of 25); white
solid. ½a 2D6
ꢁ
+14.1° (c 2.55, CHCl3). 1H NMR (CDCl3, 400 MHz) d
0.74 (3H, s, CH3-24), 0.78 (each 3H, s, CH3-23, 25), 0.96 (3H, s,
CH3-27), 1.01 (3H, s, CH3-26), 1.13, 1.16 (each 3H, s, 28-O-dimeth-
ylglutaryl CH3), 1.30, 1.31 (each 3H, s, 3-O-dimethylsuccinyl CH3),

J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
6463
(positive) m/z 1164.6608 [M+Na]+ (calcd for C67H91N5O11Na,
1164.6613).
127.1 (benzhydryl C-20, 60), 127.7 (benzyhydryl C-40), 128.4
(benzhydryl C-30, 50), 135.3 (AZT-C-6), 140.3 (benzhydryl C-10),
150.0 (AZT-C-2), 163.5 (AZT-C-4), 170.7 (28-O-dimethylsuccinyl
400-COOꢂFZN), 170.8 (3-O-dimethylsuccinyl 10-COOꢂ), 175.4
(3-O-dimethylsuccinyl 40-COOꢂ), 176.9 (28-O-dimethylsuccinyl
100-COO–). HRESIMS (positive) m/z 1138.6445 [M+Na]+ (calcd for
5.2.7.6.
1-(30-Azido-30-deoxythymidine-50-yl)-4-[3-O-(40-benz-
hydryl-30,30-dimethylsuccinyl)-lup-28-yl] 2,2-dimethylsuccinate
(41) and 4-(30-Azido-30-deoxythymidine-50-yl)-1-[3-O-(40-benz-
hydryl-30,30-dimethylsuccinyl)-lup-28-yl] 2,2-dimethylsuccinate
(42). Total yield 36.0% (starting from 285 mg of mixture of 27 and
28); used for next reaction without purification. For spectroscopic
analysis, a small amount of sample was further purified by prepara-
tive HPLC (cholester, CH3OH/H2O = 97:3); white solid.
C65H89N5O11Na, 1138.6456).
5.2.7.7. 30-Azido-30-deoxythymidine-50-yl 3-O-(40-benzhydryloxy-
30,30-dimethylsuccinyl)-lup-28-yl 3,3-dimethylglutarate (43).
Yield 66.1% (starting from 118 mg of 29); white solid. ½a D26
ꢂ0.32°
ꢁ
Compound 41: ½a 3D0
ꢁ
ꢂ6.7° (c 1.69, CHCl3). 1H NMR (CDCl3,
(c 1.53, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.76, 0.83 (each 3H,
d, J = 8.0 Hz, CH3-29, 30), 0.73 (3H, s, CH3-24), 0.77 (3H, s, CH3-
23), 0.78 (3H, s, CH3-25), 0.93 (3H, s, CH3-27), 1.00 (3H, s, CH3-
26), 1.11, 1.14 (each 3H, s, 28-O-dimethylglutaryl CH3), 1.29, 1.30
(each 3H, s, 3-O-dimethylsuccinyl CH3), 1.94 (3H, br s, AZT-CH3),
2.31–2.38, 2.43–2.50 (2H, m, AZT-H2-20), 2.41, 2.48 (each 1H, d,
J = 14.4 Hz, 28-O-dimethylglutaryl H2-200), 2.47, 2.55 (each 1H, d,
J = 15.2 Hz, 28-O-dimethylglutaryl H2-400), 2.63, 2.68 (each 1H, d,
J = 16.0 Hz, 3-O-dimethylsuccinyl H2-20), 3.77, 4.24 (each 1H, d,
J = 10.8 Hz, H2-28), 4.05 (1H, q-like, AZT-H-40), 4.19–4.23 (1H, m,
AZT-H-30), 4.25 (1H, dd, J = 4.4, 12.0 Hz, AZT-H-5a0), 4.43 (1H, dd,
J = 4.4, 12.0 Hz, AZT-H-5b0), 4.43 (1H, dd, J = 4.4, 12.0 Hz, H-3),
6.14 (1H, t-like, J = 6.4 Hz, AZT-H-10), 6.85 (1H, s, CH(Ph)2), 7.23–
7.31 (11H, m, aromatic-H and AZT-H-6), 8.48 (1H, br s, AZT-NH);
13C NMR (CDCl3, 100 MHz) d 12.6 AZT-CH3, 14.6 (C-27), 14.9
(C-29), 16.0 (C-26, 25), 16.5 (C-24), 18.1 (C-6), 20.8 (C-11), 21.6
(C-21), 22.9 (C-30), 23.5 (C-2), 25.2, 25.4 (3-O-dimethylsuccinyl
CH3), 26.8 (C-12), 26.9 (C-15), 27.9 (28-O-dimethylglutaryl CH3),
28.0 (C-23), 29.4 (C-20), 29.9 (C-16), 32.6 (28-O-dimethylglutaryl
C-300), 34.2 (C-7), 34.7 (C-22), 37.0 (C-10), 37.1 (C-13), 37.5 (AZT-
C-20), 37.7 (C-4), 38.3 (C-1), 40.7 (3-O-dimethylsuccinyl C-30), 40.9
(C-8), 42.8 (C-14), 44.5 (3-O-dimethylsuccinyl C-20, 28-O-dimethyl-
glutaryl C-400, C-19), 45.0 (28-O-dimethylsuccinyl C-200), 46.4 (C-17),
48.1 (C-18), 49.9 (C-9), 55.3 (C-5), 60.4 (AZT-C-30), 62.7 (C-28, AZT-
C-50), 77.0 (CH(Ph)2), 81.3 (C-3), 81.8 (AZT-C-40), 85.2 (AZT-C-10),
111.3 (AZT-C-5), 127.0, 127.1 (benzhydryl C-20, 60), 127.7 (ben-
zyhydryl C-40), 128.4 (benzhydryl C-30, 50), 135.2 (AZT-C-6), 140.3
(benzhydryl C-10), 149.9 (AZT-C-2), 163.2 (AZT-C-4), 170.8
(3-O-dimethylsuccinyl 10-COOꢂ), 171.2 (28-O-dimethylglutaryl
500-COO-AZT), 172.3 (28-O-dimethylglutaryl 100-COOꢂ), 175.4 (3-
O-dimethylsuccinyl 40-COOꢂ). HRESIMS (positive) m/z 1130.6781
[M+H]+ (calcd for C66H92N5O11, 1130.6793).
400 MHz) d 0.77, 0.84 (each 3H, d, J = 8.0 Hz, CH3-29, 30), 0.74 (3H,
s, CH3-24), 0.78 (3H, s, CH3-23), 0.79 (3H, s, CH3-25), 0.94 (3H, s,
CH3-27), 1.01 (3H, s, CH3-26), 1.31 (12H, s, 3-O, 28-O-dimethylsucci-
nyl CH3), 1.94 (3H, br s, AZT-CH3), 2.25–2.32, 2.44–2.50 (each 1H, m,
AZT-H2-20), 2.65, 2.70 (each 1H, d, J = 16.0 Hz, 3-O-dimethylsuccinyl
H2-20), 2.59, 2.75 (each 1H, d, J = 16.4 Hz, 28-O-dimethylsuccinyl
H2-200), 3.81, 4.28 (each 1H, d, J = 11.2 Hz, H2-28), 4.09 (1H, q-like,
AZT-H-40), 4.25–4.29 (1H, m, AZT-H-30), 4.29 (1H, dd, J = 4.4,
12.0 Hz, AZT-H-5a0), 4.52 (1H, dd, J = 4.0, 12.0 Hz, AZT-H-5b0), 4.44
(1H, dd, J = 5.2, 10.8 Hz, H-3), 6.18 (1H, t-like, J = 6.4 Hz, AZT-H-10),
6.86 (1H, s, CH(Ph)2), 7.24–7.33 (11H, m, aromatic-H and AZT-H-
6), 8.59 (1H, br s, AZT-NH); 13C NMR (CDCl3, 100 MHz) d 12.4 AZT-
CH3, 14.6 (C-27), 14.8 (C-29), 15.9 (C-26), 16.0 (C-25), 16.5 (C-24),
18.1 (C-6), 20.7 (C-11), 21.5 (C-21), 22.9 (C-30), 23.5 (C-2), 25.2,
25.4 (3-O-dimethylsuccinyl CH3), 25.0, 25.9 (28-O-dimethylsuccinyl
CH3), 26.8 (C-12), 26.8 (C-15), 27.9 (C-23), 29.4 (C-20), 29.8 (C-16),
34.1 (C-7), 34.6 (C-22), 36.9 (C-10), 37.1 (C-13), 37.4 (AZT-C-20),
37.6 (C-4), 38.3 (C-1), 40.7 (dimethylsuccinyl C-30, C-300), 40.9 (C-
8), 42.8 (C-14), 44.5 (3-O-dimethylsuccinyl C-20, C-19), 44.6 (28-O-
dimethylsuccinyl C-200), 46.5 (C-17), 48.1 (C-18), 49.9 (C-9), 55.3
(C-5), 60.4 (AZT-C-30), 63.1 (C-28), 63.3 (AZT-C-50), 77.0 (CH(Ph)2),
81.3 (C-3), 81.9 (AZT-C-40), 84.8 (AZT-C-10), 111.4 (AZT-C-5), 127.0,
127.1 (benzhydryl C-20, 60), 127.7 (benzyhydryl C-40), 128.4
(benzhydryl C-30, 50), 135.0 (AZT-C-6), 140.3 (benzhydryl C-10),
150.0 (AZT-C-2), 163.3 (AZT-C-4), 170.9 (3-O-dimethylsuccinyl
10-COOꢂ), 171.7 (28-O-dimethylsuccinyl 100-COOꢂ), 175.4 (3-O-
dimethylsuccinyl 40-COOꢂ), 176.5 (28-O-dimethylsuccinyl 400-
COO-AZT). HRESIMS (positive) m/z 1116.6604 [M+H]+ (calcd for
C65H90N5O11, 1116.6637).
Compound 42: ½a 3D0
ꢁ
ꢂ1.8° (c 2.69, CHCl3). 1H NMR (CDCl3,
400 MHz) d 0.77, 0.84 (each 3H, d, J = 8.0 Hz, CH3-29, 30), 0.74
(3H, s, CH3-24), 0.78 (3H, s, CH3-23), 0.80 (3H, s, CH3-25), 0.94
(3H, s, CH3-27), 1.02 (3H, s, CH3-26), 1.31 (6H, s, 3-O-dim-
ethylsuccinyl CH3), 1.31, 1.34 (each 3H, s, 28-O-dimethylsuccinyl
CH3), 1.96 (3H, br s, AZT-CH3), 2.32–2.39, 2.44–2.50 (each 1H, m,
AZT-H2-20), 2.65, 2.70 (each 1H, d, J = 16.0 Hz, 3-O-dimethylsucci-
nyl H2-20), 2.61, 2.66 (each 1H, d, J = 16.0 Hz, 28-O-dimethylsucci-
nyl H2-300), 3.81, 4.29 (each 1H, d, J = 10.8 Hz, H2-28), 4.04 (1H,
q-like, AZT-H-40), 4.18–4.22 (1H, m, AZT-H-30), 4.26 (1H, dd,
J = 3.6, 12.4 Hz, AZT-H-5a0), 4.42 (1H, dd, J = 4.4, 12.4 Hz, AZT-H-
5b0), 4.44 (1H, dd, J = 5.2, 10.8 Hz, H-3), 6.15 (1H, t-like, J = 6.4 Hz,
AZT-H-10), 6.86 (1H, s, CH(Ph)2), 7.24–7.33 (11H, m, aromatic-H
and AZT-H-6), 8.93 (1H, br s, AZT-NH); 13C NMR (CDCl3,
100 MHz) d 12.5 AZT-CH3, 14.6 (C-27), 14.9 (C-29), 16.0 (C-26,
25), 16.5 (C-24), 18.1 (C-6), 20.7 (C-11), 21.5 (C-21), 22.9 (C-30),
23.5 (C-2), 25.2, 25.4 (3-O-dimethylsuccinyl CH3), 25.4, 25.8 (28-
O-dimethylsuccinyl CH3), 26.8 (C-12), 26.9 (C-15), 27.9 (C-23),
29.4 (C-20), 29.9 (C-16), 34.1 (C-7), 34.6 (C-22), 36.9 (C-10), 37.1
(C-13), 37.5 (AZT-C-20, 37.6 (C-4), 38.3 (C-1), 40.7 (3-O-dim-
ethylsuccinyl C-30), 40.9 (C-8), 41.0 (28-O-dimethylsuccinyl C-200),
42.8 (C-14), 43.8 (28-O-dimethylsuccinyl C-300), 44.5 (3-O-dim-
ethylsuccinyl C-20, C-19), 46.7 (C-17), 48.1 (C-18), 49.9 (C-9), 55.3
(C-5), 60.2 (AZT-C-30), 62.8 (AZT-C-50), 63.3 (C-28), 77.0 (CH(Ph)2),
81.3 (C-3), 81.7 (AZT-C-40), 85.1 (AZT-C-10), 111.3 (AZT-C-5), 127.0,
5.2.7.8.
1-(30-Azido-30-deoxythymidine-50-yl)-4-[3-O-(40-ben-
zhydryloxyglutaryl)-lup-28-yl] 2,2-dimethylsuccinate (44) and
4-(30-Azido-30-deoxythymidine-50-yl)-1-[3-O-(40-benzhydryloxy-
glutaryl)-lup-28-yl] 2,2-dimethylsuccinate (45). Yield 15.8% and
10.5%, respectively (starting from 208 mg of mixture of 30 and
31); separated by preparative HPLC (cholester, MeOH/
H2O = 97:3); colorless oil.
Compound 44: ½a 2D7
ꢁ
ꢂ1.96° (c 0.72, CHCl3). 1H NMR (CDCl3,
400 MHz) d 0.77, 0.85 (each 3H, d, J = 8.0 Hz, CH3-29, 30), 0.83
(3H, s, CH3-24), 0.84 (3H, s, CH3-23), 0.86 (3H, s, CH3-25), 0.96
(3H, s, CH3-27), 1.03 (3H, s, CH3-26), 1.31, 1.32 (each 3H, s, 28-O-
dimethylsuccinyl CH3), 1.94 (3H, br s, AZT-CH3), 2.00 (2H, quint,
J = 7.2 Hz, 3-O-glutaryl H2-30), 2.26–2.35, 2.44–2.50 (each 1H, m,
AZT-H2-20), 2.35 (2H, t, J = 7.2 Hz, 3-O-glutaryl H2-20), 2.50 (2H, t,
J = 7.2 Hz, 3-O-glutaryl H2-40), 2.60, 2.74 (each 1H, d, J = 16.0 Hz,
28-O-dimethylsuccinyl H2-200), 3.82, 4.28 (each 1H, d, J = 10.8 Hz,
H2-28), 4.09 (1H, q-like, AZT-H-40), 4.25–4.30 (1H, m, AZT-H-30),
4.28 (1H, dd, J = 4.0, 12.0 Hz, AZT-H-5a0), 4.51 (1H, dd, J = 4.0,
12.0 Hz, AZT-H-5b0), 4.49 (1H, dd, J = 6.0, 10.0 Hz, H-3), 6.17 (1H,
t-like, J = 6.4 Hz, AZT-H-10), 6.89 (1H, s, CH(Ph)2), 7.25–7.34 (11H,
m, aromatic-H and AZT-H-6), 8.71 (1H, br s, AZT-NH); 13C NMR
(CDCl3, 100 MHz) d 12.4 AZT-CH3, 14.6 (C-27), 14.9 (C-29), 16.0

6464
J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
(C-26), 16.1 (C-25), 16.5 (C-24), 18.2 (C-6), 20.4 (3-O-glutaryl C-30),
20.8 (C-11), 21.6 (C-21), 22.9 (C-30), 23.7 (C-2), 25.1, 25.8 (28-O-
dimethylsuccinyl CH3), 26.8 (C-12), 26.9 (C-15), 28.0 (C-23), 29.4
(C-20), 29.9 (C-16), 33.6 (3-O-glutaryl C-40), 33.7 (3-O-glutaryl C-
20), 34.2 (C-7), 34.6 (C-22), 37.0 (C-10), 37.2 (C-13), 37.5 (AZT-C-
20), 37.8 (C-4), 38.4 (C-1), 40.7 (28-O-dimethylsuccinyl C-300), 40.9
(C-8), 42.9 (C-14), 44.5 (C-19), 44.6 (28-O-dimethylsuccinyl C-
200), 46.5 (C-17), 48.1 (C-18), 49.9 (C-9), 55.3 (C-5), 60.4 (AZT-C-
30), 63.1 (C-28), 63.3 (AZT-C-50), 76.9 (CH(Ph)2), 81.0 (C-3), 82.0
(AZT-C-40), 84.9 (AZT-C-10), 111.4 (AZT-C-5), 127.0 (benzhydryl C-
20, 60), 127.9 (benzyhydryl C-40), 128.5 (benzhydryl C-30, 50), 135.0
(AZT-C-6), 140.2 (benzhydryl C-10), 150.0 (AZT-C-2), 163.4 (AZT-
C-4), 171.7 (28-O-dimethylsuccinyl 100-COOꢂ), 171.9 (3-O-glutaryl
50-COOꢂ), 172.6 (3-O-glutaryl 10-COOꢂ), 176.5 (28-O-dim-
ethylsuccinyl 400-COO-AZT). HRESIMS (positive) m/z 1102.6483
[M+H]+ (calcd for C64H88N5O11, 1102.6480).
20.8 (C-11), 21.6 (C-21), 22.8 (C-30), 23.7 (C-2), 26.8 (C-12), 26.9
(C-15), 28.0 (C-23), 29.4 (C-20), 29.9 (C-16), 33.1 (28-O-glutaryl
C-400), 33.2 (28-O-glutaryl C-200), 33.6 (3-O-glutaryl C-40), 33.7 (3-
O-glutaryl C-20), 34.2 (C-7), 34.6 (C-22), 37.0 (C-10), 37.2 (C-13),
37.4 (AZT-C-20), 37.8 (C-4), 38.3 (C-1), 40.9 (C-8), 42.8 (C-14),
44.5 (C-19), 46.5 (C-17), 48.1 (C-18), 49.9 (C-9), 55.3 (C-5), 60.6
(AZT-C-30), 62.9 (C-28), 63.3 (AZT-C-50), 76.8 (CH(Ph)2), 81.0 (C-
3), 81.7 (AZT-C-40), 85.6 (AZT-C-10), 111.3 (AZT-C-5), 127.0 (benz-
hydryl C-20, 60), 127.8 (benzyhydryl C-40), 128.4 (benzhydryl C-30,
50), 135.3 (AZT-C-6), 140.2 (benzhydryl C-10), 150.0 (AZT-C-2),
163.5 (AZT-C-4), 171.8 (3-O-glutaryl 50-COOꢂ), 172.3 (28-O-gluta-
ryl 500-COO-AZT), 172.6 (3-O-glutaryl 10-COOꢂ), 173.1 (28-O-gluta-
ryl 100-COOꢂ). HRESIMS (positive) m/z 1112.6271 [M+Na]+ (calcd
for C63H87N5O11Na, 1112.6300).
5.2.7.10. 30-Azido-30-deoxythymidine-50-yl 3-O-(40-benzhydryl-
oxyglutaryl)-lup-28-yl 3,3-dimethylglutarate (47). Yield 48.7%
Compound 45: ½a 2D7
ꢁ
+1.4° (c 0.99, CHCl3). 1H NMR (CDCl3,
400 MHz) d 0.77, 0.85 (each 3H, d, J = 8.0 Hz, CH3-29, 30), 0.83
(3H, s, CH3-24), 0.84 (3H, s, CH3-23), 0.86 (3H, s, CH3-25), 0.96
(3H, s, CH3-27), 1.05 (3H, s, CH3-26), 1.31, 1.34 (each 3H, s, 28-O-
dimethylsuccinyl CH3), 1.96 (3H, br s, AZT-CH3), 2.00 (2H, quint,
J = 7.2 Hz, 3-O-glutaryl H2-30), 2.33–2.52 (2H, m, AZT-H2-20), 2.35
(2H, t, J = 7.2 Hz, 3-O-glutaryl H2-20), 2.50 (2H, t, J = 7.2 Hz, 3-O-
glutaryl H2-40), 2.62, 2.66 (each 1H, d, J = 16.0 Hz, 28-O-dim-
ethylsuccinyl H2-200), 3.82, 4.30 (each 1H, d, J = 10.8 Hz, H2-28),
4.05 (1H, q-like, AZT-H-40), 4.17–4.22 (1H, m, AZT-H-30), 4.26
(1H, dd, J = 3.6, 12.0 Hz, AZT-H-5a0), 4.42 (1H, dd, J = 4.0, 12.0 Hz,
AZT-H-5b0), 4.49 (1H, dd, J = 6.0, 10.0 Hz, H-3), 6.15 (1H, t-like,
J = 6.4 Hz, AZT-H-10), 6.89 (1H, s, CH(Ph)2), 7.25–7.34 (11H, m, aro-
matic-H and AZT-H-6), 8.74 (1H, br s, AZT-NH); 13C NMR (CDCl3,
100 MHz) d 12.5 AZT-CH3, 14.6 (C-27), 14.9 (C-29), 16.0 (C-26),
16.1 (C-25), 16.5 (C-24), 18.1 (C-6), 20.4 (3-O-glutaryl C-30), 20.8
(C-11), 21.6 (C-21), 22.9 (C-30), 23.7 (C-2), 25.5, 25.8 (28-O-dim-
ethylsuccinyl CH3), 26.8 (C-12), 26.9 (C-15), 28.0 (C-23), 29.4 (C-
20), 29.9 (C-16), 33.6 (3-O-glutaryl C-40), 33.7 (3-O-glutaryl C-20),
34.2 (C-7), 34.7 (C-22), 37.0 (C-10), 37.2 (C-13), 37.5 (AZT-C-20),
37.8 (C-4), 38.4 (C-1), 40.9 (C-8), 41.0 (28-O-dimethylsuccinyl C-
200), 42.9 (C-14), 43.8 (28-O-dimethylsuccinyl C-300), 44.6 (C-19),
46.7 (C-17), 48.2 (C-18), 50.0 (C-9), 55.3 (C-5), 60.3 (AZT-C-30),
62.9 (AZT-C-50), 63.3 (C-28), 76.9 (CH(Ph)2), 81.0 (C-3), 81.8
(AZT-C-40), 85.2 (AZT-C-10), 111.3 (AZT-C-5), 127.0 (benzhydryl C-
20, 60), 127.9 (benzyhydryl C-40), 128.5 (benzhydryl C-30, 50),
135.3 (AZT-C-6), 140.2 (benzhydryl C-10), 150.0 (AZT-C-2), 163.4
(AZT-C-4), 170.7 (28-O-dimethylsuccinyl 400-COO-AZT), 171.9 (3-
O-glutaryl 50-COOꢂ), 172.6 (3-O-glutaryl 10-COOꢂ), 176.8 (28-O-
dimethylsuccinyl 100-COOꢂ). HRESIMS (positive) m/z 1102.6475
[M+H]+ (calcd for C64H88N5O11, 1102.6480).
(starting from 76 mg of 33); colorless oil. ½a D27
ꢁ
+4.1° (c 0.54, CHCl3).
1H NMR (CDCl3, 400 MHz) d 0.78, 0.84 (each 3H, d, J = 8.0 Hz, CH3-
29, 30), 0.83 (3H, s, CH3-24), 0.84 (3H, s, CH3-23), 0.86 (3H, s, CH3-
25), 0.96 (3H, s, CH3-27), 1.04 (3H, s, CH3-26), 1.12, 1.15 (each 3H,
s, 28-O-dimethylglutaryl CH3), 1.95 (3H, d, J = 1.2 Hz, AZT-CH3),
2.00 (2H, quint, J = 7.2 Hz, 3-O-glutaryl H2-30), 2.32–2.52 (2H, m,
AZT-H2-20), 2.35 (2H, t, J = 7.2 Hz, 3-O-glutaryl H2-20), 2.43, 2.49
(each 1H, d, J = 14.4 Hz, 28-O-dimethylglutaryl H2-200), 2.48, 2.56
(each 1H, d, J = 14.4 Hz, 28-O-dimethylglutaryl H2-400), 2.50 (2H, t,
J = 7.2 Hz, 3-O-glutaryl H2-40), 3.79, 4.27 (each 1H, d, J = 11.2 Hz,
H2-28), 4.07 (1H, q-like, AZT-H-40), 4.20–4.28 (1H, m, AZT-H-30),
4.26 (1H, dd, J = 3.6, 12.0 Hz, AZT-H-5a0), 4.43 (1H, dd, J = 4.4,
12.0 Hz, AZT-H-5b0), 4.49 (1H, dd, J = 6.0, 10.4 Hz, H-3), 6.15 (1H,
t-like, J = 6.4 Hz, AZT-H-10), 6.89 (1H, s, CH(Ph)2), 7.25–7.33 (11H,
m, aromatic-H and AZT-H-6), 8.91 (1H, br s, AZT-NH); 13C NMR
(CDCl3, 100 MHz) d 12.5 AZT-CH3, 14.6 (C-27), 14.8 (C-29), 16.0
(C-26, C-25), 16.5 (C-24), 18.1 (C-6), 20.3 (3-O-glutaryl C-30), 20.8
(C-11), 21.6 (C-21), 22.8 (C-30), 23.7 (C-2), 26.8 (C-12), 26.9
(C-15), 27.8, 27.9 (28-O-dimethylglutaryl CH3), 28.0 (C-23), 29.4
(C-20), 29.9 (C-16), 32.5 (28-O-dimethylglutaryl C-300), 33.6 (3-O-
glutaryl C-40), 33.7 (3-O-glutaryl C-20), 34.2 (C-7), 34.7 (C-22),
37.0 (C-10), 37.2 (C-13), 37.5 (AZT-C-20), 37.8 (C-4), 38.3 (C-1),
40.9 (C-8), 42.8 (C-14), 44.5 (28-O-dimethylglutaryl C-400 and C-
19), 45.1 (28-O-dimethylglutaryl C-200), 46.4 (C-17), 48.1 (C-18),
49.9 (C-9), 55.3 (C-5), 60.5 (AZT-C-30), 62.7 (C-28, and AZT-C-50),
76.9 (CH(Ph)2), 81.0 (C-3), 81.8 (AZT-C-40), 85.2 (AZT-C-10), 111.3
(AZT-C-5), 127.0 (benzhydryl C-20, 60), 127.9 (benzyhydryl C-40),
128.5 (benzhydryl C-30, 50), 135.2 (AZT-C-6), 140.2 (benzhydryl
C-10), 150.1 (AZT-C-2), 163.5 (AZT-C-4), 171.1 (28-O-dimethylglut-
aryl 500-COO-AZT), 171.8 (3-O-glutaryl 50-COOꢂ), 172.2 (28-O-dim-
ethylglutaryl 100-COOꢂ), 172.6 (3-O-glutaryl 10-COOꢂ). HRESIMS
(positive) m/z 1138.6449 [M+Na]+ (calcd for C65H89N5O11Na,
1138.6456).
5.2.7.9. 30-Azido-30-deoxythymidine-50-yl 3-O-(40-benzhydryl-
oxyglutaryl)-lup-28-yl glutarate (46). Yield 49.8% (starting from
79 mg of 32); colorless oil. ½a D27
ꢁ
+4.4° (c 0.93, CHCl3). 1H NMR
(CDCl3, 400 MHz) d 0.78, 0.84 (each 3H, d, J = 8.0 Hz, CH3-29, 30),
0.83 (each 3H, s, CH3-24, CH3-23), 0.86 (3H, s, CH3-25), 0.96 (3H,
s, CH3-27), 1.05 (3H, s, CH3-26), 1.94 (3H, d, J = 1.2 Hz, AZT-CH3),
1.99 (4H, quint, J = 7.2 Hz, 3-O, 28-O-glutaryl H2-30), 2.32–2.52
(2H, m, AZT-H2-20), 2.35 (2H, t, J = 7.2 Hz, 3-O-glutaryl H2-20),
2.41 (2H, t, J = 7.2 Hz, 28-O-glutaryl H2-200), 2.46 (2H, t, J = 7.2 Hz,
28-O-glutaryl H2-400), 2.50 (2H, t, J = 7.2 Hz, 3-O-glutaryl H2-40),
3.83, 4.30 (each 1H, d, J = 11.2 Hz, H2-28), 4.07 (1H, q-like, AZT-
H-40), 4.20–4.24 (1H, m, AZT-H-30), 4.32 (1H, dd, J = 4.0, 12.0 Hz,
AZT-H-5a0), 4.40 (1H, dd, J = 4.4, 12.0 Hz, AZT-H-5b0), 4.49 (1H,
dd, J = 6.0, 10.4 Hz, H-3), 6.10 (1H, t-like, J = 6.4 Hz, AZT-H-10),
6.89 (1H, s, CH(Ph)2), 7.25–7.33 (11H, m, aromatic-H and AZT-H-
6), 9.04 (1H, br s, AZT-NH); 13C NMR (CDCl3, 100 MHz) d 12.5
AZT-CH3, 14.6 (C-27), 14.8 (C-29), 16.0 (C-26, C-25), 16.5 (C-24),
18.1 (C-6), 20.0 (28-O-glutaryl C-300), 20.3 (3-O-glutaryl C-30),
5.2.7.11. 30-Azido-30-deoxythymidine-50-yl 3-O-(40-benzhydryl-
oxy-30,30-dimethylglutaryl)-lup-28-yl 3,3-dimethylglutarate (48).
Yield 68.3% (starting from 126 mg of 34); colorless oil. ½a D26
ꢁ
+0.97°
(c 2.37, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.78, 0.84 (each 3H,
d, J = 8.0 Hz, CH3-29, 30), 0.81 (3H, s, CH3-24), 0.83 (6H, s, CH3-23,
CH3-25), 0.95 (3H, s, CH3-27), 1.03 (3H, s, CH3-26), 1.09 (6H, s, 3-
O-dimethylglutaryl CH3), 1.12, 1.15 (each 3H, s, 28-O-dimethylglut-
aryl CH3), 1.95 (3H, br s, AZT-CH3), 2.32–2.52 (2H, m, AZT-H2-20),
2.35, 2.42 (each 1H, d, J = 14.4 Hz, 3-O-dimethylglutaryl H2-20),
2.43, 2.49 (each 1H, d, J = 14.4 Hz, 28-O-dimethylglutaryl H2-200),
2.48, 2.56 (each 1H, d, J = 14.4 Hz, 28-O-dimethylglutaryl H2-400),
2.55, 2.58 (each 1H, d, J = 14.4 Hz, 3-O-dimethylglutaryl H2-40),
3.79, 4.26 (each 1H, d, J = 11.2 Hz, H2-28), 4.07 (1H, q-like, AZT-H-
40), 4.21–4.28 (1H, m, AZT-H-30), 4.26 (1H, dd, J = 3.6, 12.4 Hz,
AZT-H-5a0), 4.44 (1H, dd, J = 4.4, 12.4 Hz, AZT-H-5b0), 4.47 (1H, dd,

J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
6465
J = 4.8, 10.8 Hz, H-3), 6.15 (1H, t-like, J = 6.4 Hz, AZT-H-10), 6.89 (1H,
Compound 50: ½a 2D9
ꢁ
+8.2° (c 2.38, CHCl3). 1H NMR (CDCl3,
s, CH(Ph)2), 7.22–7.34 (11H, m, aromatic-H and AZT-H-6), 8.97 (1H,
br s, AZT-NH); 13C NMR (CDCl3, 100 MHz) d 12.6 AZT-CH3, 14.6
(C-27), 14.9 (C-29), 16.0 (C-26, C-25), 16.6 (C-24), 18.1 (C-6), 20.7
(C-11), 21.6 (C-21), 22.9 (C-30), 23.7 (C-2), 26.8 (C-12), 26.9
(C-15), 27.6 (3-O-dimethylglutaryl CH3), 27.9, 28.0 (28-O-dimethyl-
glutaryl CH3), 28.0 (C-23), 29.4 (C-20), 29.9 (C-16), 32.6 (28-O-dim-
ethylglutaryl C-300), 32.8 (3-O-dimethylglutaryl C-30), 34.2 (C-7),
34.7 (C-22), 37.0 (C-10), 37.1 (C-13), 37.5 (AZT-C-20), 37.6 (C-4),
38.3 (C-1), 40.9 (C-8), 42.8 (C-14), 44.5 (28-O-dimethylglutaryl
C-400 and C-19), 45.0 (28-O-dimethylglutaryl C-200), 45.5 (3-O-gluta-
ryl C-40), 45.8 (3-O-glutaryl C-20), 46.4 (C-17), 48.1 (C-18), 49.9 (C-
9), 55.3 (C-5), 60.4 (AZT-C-30), 62.7 (C-28, and AZT-C-50), 76.7
(CH(Ph)2), 80.9 (C-3), 81.8 (AZT-C-40), 85.2 (AZT-C-10), 111.3 (AZT-
C-5), 127.1 (benzhydryl C-20, 60), 127.8 (benzyhydryl C-40), 128.4
(benzhydryl C-30, 50), 135.3 (AZT-C-6), 140.2 (benzhydryl C-10),
150.0 (AZT-C-2), 163.6 (AZT-C-4), 170.8 (3-O-dimethylglutaryl
50-COOꢂ), 171.2 (28-O-dimethylglutaryl 500-COO-AZT), 171.6
(3-O-dimethylglutaryl 10-COOꢂ), 172.3 (28-O-dimethylglutaryl 100-
COOꢂ). HRESIMS (positive) m/z 1144.6917 [M+H]+ (calcd for
400 MHz) d 0.81 (3H, s, CH3-24), 0.83 (each 3H, s, CH3-23, 25),
0.96 (3H, s, CH3-27), 1.00 (3H, s, CH3-26), 1.29, 1.30 (each 3H, s, 3-
O-dimethylsuccinyl CH3), 1.30, 1.31 (each 3H, s, 28-O-dim-
ethylsuccinyl CH3), 1.68 (3H, s, CH3-30), 1.93 (3H, br s, AZT-CH3),
2.26–2.33, 2.44–2.50 (each 1H, m, AZT-H2-20), 2.37–2.44 (1H, m,
H-19), 2.56, 2.68 (each 1H, d, J = 16.0 Hz, 3-O-dimethylsuccinyl
H2-20), 2.60, 2.74 (each 1H, d, J = 16.4 Hz, 28-O-dimethylsuccinyl
H2-200), 3.84, 4.27 (each 1H, d, J = 11.2 Hz, H2-28), 4.09 (1H, q-like,
AZT-H-40), 4.25–4.31 (2H, m, AZT-H-30 and AZT-H-5a0), 4.51 (1H,
dd, J = 4.0, 12.0 Hz, AZT-H-5b0), 4.48 (1H, dd, J = 5.6, 11.2 Hz, H-3),
4.59, 4.68 (each 1H, br s, H2-29), 6.16 (1H, t-like, J = 6.4 Hz, AZT-H-
10), 7.28 (1H, d, J = 0.8 Hz, AZT-H-6), 9.38 (1H, br s, AZT-NH); 13C
NMR (CDCl3, 100 MHz) d 12.4 AZT-CH3, 14.7 (C-27), 15.9 (C-26),
16.1 (C-25), 16.5 (C-24), 18.1 (C-6), 19.1 (C-30), 20.7 (C-11), 23.6
(C-2), 25.1 (C-12), 25.1, 25.6 (3-O-dimethylsuccinyl CH3), 25.1,
25.9 (28-O-dimethylsuccinyl CH3), 27.0 (C-15), 27.9 (C-23), 29.5
(C-21), 29.7 (C-16), 34.1 (C-7), 34.5 (C-22), 37.0 (C-10), 37.5 (AZT-
C-20), 37.6 (C-13), 37.7 (C-4), 38.4 (C-1), 40.5 (3-O-dimethylsuccinyl
C-30), 40.7 (28-O-dimethylsuccinyl C-300), 40.8 (C-8), 42.7 (C-14),
44.6 (28-O-dimethylsuccinyl C-200), 44.7 (3-O-dimethylsuccinyl
C-20), 46.3 (C-17), 47.7 (C-19), 48.7 (C-18), 50.2 (C-9), 55.4 (C-5),
60.4 (AZT-C-30), 63.1 (C-28), 63.0 (AZT-C-50), 81.5 (C-3), 82.0
(AZT-C-40), 84.9 (AZT-C-10), 110.0 (C-29), 111.3 (AZT-C-5), 135.2
(AZT-C-6), 149.9 (C-20), 150.1 (AZT-C-2), 164.0 (AZT-C-4), 171.1
(3-O-dimethylsuccinyl 10-COOꢂ), 171.7 (28-O-dimethylsucci-
nyl 100-COOꢂ), 176.5 (28-O-dimethylsuccinyl 400-COO-AZT), 182.2
(3-O-dimethylsuccinyl 40-COOY). HRESIMS (positive) m/z
970.5511 [M+Na]+ (calcd for C52H77N5O11Na, 970.5517).
C67H94N5O11, 1144.6950).
5.2.8. General procedure for removing benzhydryl group (49–62)
The final products were prepared by refluxing of a solution of
benzdryl ester derivative in 80% HOAc (2–6 mL) overnight. After
cooling to room temperature, the reaction mixture was extracted
with CHCl3, and the organic layer was washed with water and
brine, dried over Na2SO4 and concentrated. The residue was puri-
fied by preparative HPLC (cholester, MeOH/H2O/HOAc = 93:6:1).
Compound 51: ½a 2D7
ꢁ
+15.6° (c 4.68, CHCl3). 1H NMR (CDCl3,
5.2.8.1. 30-Azido-30-deoxythymidine-50-yl 3-O-(30,30-dimethyl-
succinyl)-lup-20(29)en-28-yl succinate (49). Yield 46.5% (starting
400 MHz) d 0.80 (3H, s, CH3-24), 0.82 (6H, s, CH3-23, C-25), 0.96
(3H, s, CH3-27), 1.01 (3H, s, CH3-26), 1.28, 1.29 (each 3H, s, 3-O-
dimethylsuccinyl CH3), 1.30, 1.33 (each 3H, s, 28-O-dimethylsucci-
nyl CH3), 1.67 (3H, s, CH3-30), 1.94 (3H, br s, AZT-CH3), 2.33–2.50
(3H, m, AZT-H2-20 and H-19), 2.61, 2.65 (each 1H, d, J = 16.4 Hz,
28-O-dimethylsuccinyl H2-300), 2.56, 2.67 (each 1H, d, J = 15.6 Hz,
3-O-dimethylsuccinyl H2-20), 3.83, 4.27 (each 1H, d, J = 10.8 Hz,
H2-28), 4.03 (1H, q-like, AZT-H-40), 4.18–4.22 (1H, m, AZT-H-30),
4.25 (1H, dd, J = 4.0, 12.4 Hz, AZT-H-5a0), 4.41 (1H, dd, J = 4.4,
12.4 Hz, AZT-H-5b0), 4.48 (1H, dd, J = 5.2, 10.8 Hz, H-3), 4.58, 4.67
(each 1H, br s, H2-29), 6.12 (1H, t-like, J = 6.4 Hz, AZT-H-10), 7.31
(1H, d, J = 0.8 Hz, AZT-H-6), 9.95 (1H, br s, AZT-NH); 13C NMR
(CDCl3, 100 MHz) d 12.5 AZT-CH3, 14.7 (C-27), 15.9 (C-26), 16.0
(C-25), 16.4 (C-24), 18.1 (C-6), 19.1 (C-30), 20.7 (C-11), 23.5 (C-
2), 25.1 (C-12), 24.9, 25.6 (3-O-dimethylsuccinyl CH3), 25.4, 25.7
(28-O-dimethylsuccinyl CH3), 26.9 (C-15), 27.8 (C-23), 29.5
(C-21), 29.8 (C-16), 34.0 (C-7), 34.5 (C-22), 37.0 (C-10), 37.5
(AZT-C-20 and C-13), 37.6 (C-4), 38.3 (C-1), 40.4 (3-O-dim-
ethylsuccinyl C-30), 40.8 (C-8), 41.0 (28-O-dimethylsuccinyl C-200),
42.6 (C-14), 43.8 (28-O-dimethylsuccinyl C-300), 44.7 (3-O-dim-
ethylsuccinyl C-20), 46.5 (C-17), 47.7 (C-19), 48.8 (C-18), 50.2 (C-
9), 55.3 (C-5), 60.2 (AZT-C-30), 62.9 (AZT-C-50), 63.2 (C-28), 81.5
(C-3), 81.8 (AZT-C-40), 85.3 (AZT-C-10), 109.9 (C-29), 111.2
(AZT-C-5), 135.6 (AZT-C-6), 149.9 (C-20), 150.2 (AZT-C-2), 164.4
(AZT-C-4), 170.7 (28-O-dimethylsuccinyl 400-COO-AZT), 171.0
(3-O-dimethylsuccinyl 10-COOꢂ), 176.9 (28-O-dimethylsuccinyl
100-COOꢂ), 182.7 (3-O-dimethylsuccinyl 40-COOY). HRESIMS (posi-
tive) m/z 970.5490 [M+Na]+ (calcd for C52H77N5O11Na, 970.5517).
from 62 mg of 35); white solid. ½a D18
ꢁ
+7.2° (c 0.57, CHCl3). 1H NMR
(CDCl3, 500 MHz) d 0.80 (3H, s, CH3-24), 0.82 (3H, s, CH3-23), 0.83
(3H, s, CH3-25), 0.96 (3H, s, CH3-27), 1.00 (3H, s, CH3-26), 1.29, 1.31
(each 3H, s, 3-O-dimethylsuccinyl CH3), 1.68 (3H, s, CH3-30), 1.94
(3H, d, J = 0.9 Hz, AZT-CH3), 2.33–2.49 (3H, m, AZT-H2-20 and H-
19), 2.57, 2.68 (each 1H, d, J = 15.6 Hz, 3-O-dimethylsuccinyl H2-
20), 2.67–2.77 (4H, m, 28-O-succinyl H2-200, 300), 3.86, 4.30 (each
1H, d, J = 11.2 Hz, H2-28), 4.05 (1H, q-like, AZT-H-40), 4.21–4.25
(1H, m, AZT-H-30), 4.30 (1H, dd, J = 3.4, 12.3 Hz, AZT-H2-5a0), 4.49
(1H, dd, J = 5.2, 11.2 Hz, H-3), 4.52 (1H, dd, J = 4.3, 12.3 Hz, AZT-
H2-5b0), 4.58, 4.68 (each 1H, br s, H2-29), 6.14 (1H, t-like,
J = 6.3 Hz, AZT-H-10), 7.30 (1H, d, J = 1.2 Hz, AZT-H-6), 8.68 (1H,
br s, AZT-NH); 13C NMR (CDCl3, 125 MHz) d 12.6 AZT-CH3, 14.7
(C-27), 16.0 (C-26), 16.1 (C-25), 16.5 (C-24), 18.1 (C-6), 19.1 (C-
30), 20.8 (C-11), 23.6 (C-2), 25.1 (C-12), 25.1, 25.6 (3-O-dim-
ethylsuccinyl CH3), 27.0 (C-15), 27.9 (C-23), 29.0 (28-O-succinyl
C-200, 300), 29.5 (C-21), 29.7 (C-16), 34.1 (C-7), 34.6 (C-22), 37.0
(C-10), 37.6 (AZT-C-20 and C-13), 37.7 (C-4), 38.3 (C-1), 40.4
(dimethylsuccinyl C-30), 40.9 (C-8), 42.7 (C-14), 44.7 (3-O-dim-
ethylsuccinyl C-20), 46.4 (C-17), 47.7 (C-19), 48.7 (C-18), 50.2 (C-
9), 55.4 (C-5), 60.1 (AZT-C-30), 63.1 (AZT-C-50), 63.4 (C-28), 81.5
(C-3), 81.9 (AZT-C-40), 85.2 (AZT-C-10), 110.0 (C-29), 111.3 (AZT-
C-5), 135.4 (AZT-C-6), 149.9 (AZT-C-2), 150.0 (C-20), 163.5 (AZT-
C-4), 171.1 (3-O-dimethylsuccinyl 10-COOꢂ), 172.0 (28-O-succinyl
400-COO-AZT), 172.6 (28-O-succinyl 100-COOꢂ), 180.8 (3-O-dim-
ethylsuccinyl 40-COOY). HRESIMS (positive) m/z 942.5218
[M+Na]+ (calcd for C50H73N5O11Na, 942.5204).
5.2.8.3. 30-Azido-30-deoxythymidine-50-yl 3-O-(30,30-dimethyl-
succinyl)-lup-20(29)en-28-yl glutarate (52). Yield 42.7% (starting
5.2.8.2. 1-(30-Azido-30-deoxythymidine-50-yl)-4-[3-O-(30,30-dim-
ethylsuccinyl)-lup-20(29)en-28-yl] 2,2-dimethylsuccinate (50)
and 4-(30-Azido-30-deoxythymidine-50-yl)-1-[3-O-(30,30-dimethyl-
succinyl)-lup-20(29)en-28-yl] 2,2-dimethylsuccinate (51). Total
yield 78.9% (starting from 105 mg of mixture of 36 and 37); in a
ratio of 2:1; white solid.
from 90 mg of 38); white solid. ½a D15
ꢁ
+18.8° (c 0.6, CHCl3). 1H NMR
(CDCl3, 500 MHz) d 0.81 (3H, s, CH3-24), 0.83 (3H, s, CH3-23), 0.83
(3H, s, CH3-25), 0.96 (3H, s, CH3-27), 1.02 (3H, s, CH3-26), 1.29,
1.31 (each 3H, s, 3-O-dimethylsuccinyl CH3), 1.68 (3H, s, CH3-
30), 1.94 (3H, d, J = 1.2 Hz, AZT-CH3), 1.99 (2H, quint, J = 7.2 Hz,

6466
J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
28-O-glutaryl H2-300), 2.33–2.49 (3H, m, AZT-H2-20 and H-19), 2.42
(2H, t, J = 7.2 Hz, 28-O-glutaryl H2-200), 2.46 (2H, t, J = 7.2 Hz,
28-O-glutaryl H2-400), 2.57, 2.67 (each 1H, d, J = 15.8 Hz, 3-O-dim-
ethylsuccinyl H2-20), 3.85, 4.28 (each 1H, d, J = 10.8 Hz, H2-28),
4.07 (1H, q-like, AZT-H-40), 4.20–4.24 (1H, m, AZT-H-30), 4.32 (1H,
dd, J = 3.9, 12.3 Hz, AZT-H2-5a0), 4.40 (1H, dd, J = 4.7, 12.3 Hz, AZT-
H2-5b0), 4.49 (1H, dd, J = 5.2, 11.2 Hz, H-3), 4.59, 4.68 (each 1H, br
s, H2-29), 6.09 (1H, t-like, J = 6.3 Hz, AZT-H-10), 7.21 (1H, d,
J = 1.1 Hz, AZT-H-6); 13C NMR (CDCl3, 125 MHz) d 12.6 AZT-CH3,
14.7 (C-27), 16.0 (C-26), 16.1 (C-25), 16.5 (C-24), 18.1 (C-6), 19.1
(C-30), 20.0 (28-O-glutaryl C-300), 20.8 (C-11), 23.6 (C-2), 25.1
(C-12), 25.1, 25.6 (3-O-dimethylsuccinyl CH3), 27.0 (C-15), 27.9
(C-23), 29.5 (C-21), 29.8 (C-16), 33.1 (28-O-glutaryl C-400), 33.2
(28-O-glutaryl C-200), 34.1 (C-7), 34.6 (C-22), 37.0 (C-10), 37.5
(AZT-C-20), 37.6 (C-13), 37.7 (C-4), 38.4 (C-1), 40.4 (dimethylsucci-
nyl C-30), 40.9 (C-8), 42.7 (C-14), 44.8 (3-O-dimethylsuccinyl C-20),
46.4 (C-17), 47.7 (C-19), 48.7 (C-18), 50.2 (C-9), 55.4 (C-5),
60.6 (AZT-C-30), 63.0 (C-28), 63.3 (AZT-C-50), 81.6 (C-3), 81.7
(AZT-C-40), 85.7 (AZT-C-10), 110.0 (C-29), 111.3 (AZT-C-5), 135.5
(AZT-C-6), 149.8 (C-20), 149.9 (AZT-C-2), 163.5 (AZT-C-4), 171.2
(3-O-dimethylsuccinyl 10-COOꢂ), 172.3 (28-O-glutaryl 500-COO-
AZT), 173.1 (28-O-glutaryl 100-COOꢂ), 180.8 (3-O-dimethylsuccinyl
40-COOY). HRESIMS (positive) m/z 956.5361 [M+Na]+ (calcd for
AZT-CH3), 2.32–2.51 (3H, m, AZT-H2-20 and H-19), 2.40, 2.47
(each 1H, d, J = 14.4 Hz, 3-O-dimethylglutaryl H2-20), 2.46 (2H, s,
3-O-dimethylglutaryl H2-40), 2.44, 2.49 (each 1H, d, J = 14.4 Hz,
28-O-dimethylglutaryl H2-200), 2.48, 2.56 (each 1H, d, J = 14.4 Hz,
28-O-dimethylglutaryl H2-300), 3.81, 4.26 (each 1H, d, J = 11.2 Hz,
H2-28), 4.07 (1H, q-like, AZT-H-40), 4.20–4.28 (1H, m, AZT-H-30),
4.26 (1H, dd, J = 4.0, 12.4 Hz, AZT-H-5a0), 4.43 (1H, dd, J = 4.0,
12.4 Hz, AZT-H-5b0), 4.49 (1H, dd, J = 5.2, 11.6 Hz, H-3), 4.60, 4.69
(each 1H, br s, H2-29), 6.14 (1H, t-like, J = 6.4 Hz, AZT-H-10), 7.29
(1H, br s, AZT-H-6), 9.41 (1H, br s, AZT-NH); 13C NMR (CDCl3,
100 MHz) d 12.5 AZT-CH3, 14.7 (C-27), 16.0 (C-26), 16.1 (C-25),
16.6 (C-24), 18.1 (C-6), 19.1 (C-30), 20.8 (C-11), 23.7 (C-2), 25.1
(C-12), 27.0 (C-15), 27.9 (3-O-dimethylglutaryl CH3), 27.9, 28.0
(28-O-dimethylglutaryl CH3), 28.0 (C-23), 29.5 (C-21), 29.8 (C-16),
32.6 (28-O-dimethylglutaryl C-300, 3-O-dimethylglutaryl C-30), 34.1
(C-7), 34.6 (C-22), 37.0 (C-10), 37.5 (AZT-C-20, C-13), 37.6 (C-4),
38.3 (C-1), 40.8 (C-8), 42.7 (C-14), 44.5 (28-O-dimethylglutaryl C-
400), 45.0 (28-O-dimethylsuccinyl C-200), 45.2 (3-O-dimethylglutaryl
C-40), 45.6 (3-O-dimethylglurayl C-20), 46.2 (C-17), 47.7 (C-19), 48.7
(C-18), 50.2 (C-9), 55.4 (C-5), 60.4 (AZT-C-30), 62.6 (C-28), 62.7 (AZT-
C-50), 81.3 (C-3), 81.8 (AZT-C-40), 85.3 (AZT-C-10), 109.9 (C-29),
111.2 (AZT-C-5), 135.4 (AZT-C-6), 150.0 (C-20), 150.1 (AZT-C-2),
164.0 (AZT-C-4), 171.2 (28-O-dimethylglutaryl 500-COO-AZT),
172.3 (3-O-dimethylglutaryl 10-COOꢂ, 28-O-dimethylglutaryl 100-
COOꢂ), 176.0 (3-O-dimethylglutaryl 50-COOY). HRESIMS (positive)
m/z 998.5824 [M+Na]+ (calcd for C54H81N5O11Na, 998.5830).
C51H75N5O11Na, 956.5361).
5.2.8.4. 30-Azido-30-deoxythymidine-50-yl 3-O-(30,30-dimethyl-
succinyl)-lup-20(29)en-28-yl 3,3-dimethylglutarate (53). Yield
87.3% (starting from 114 mg of 39); white solid. ½a D27
ꢁ
+15.3° (c
5.2.8.6. 1-(30-Azido-30-deoxythymidine-50-yl)-4-[3-O-(30,30-dim-
ethylsuccinyl)-lup-28-yl] 2,2-dimethylsuccinate (55) and 4-(30-
Azido-30-deoxythymidine-50-yl)-1-[3-O-(30,30-dimethylsuccinyl)-
lup-28-yl] 2,2-dimethylsuccinate (56). Total yield 56.7% (starting
from 148 mg of mixture of 41 and 42); in a ratio of 1.1:1; white
solid.
2.84, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.80 (3H, s, CH3-24),
0.83 (each 3H, s, CH3-23, 25), 0.96 (3H, s, CH3-27), 1.01 (3H, s,
CH3-26), 1.12, 1.15 (each 3H, s, 28-O-dimethylglutaryl CH3), 1.29,
1.30 (each 3H, s, 3-O-dimethylsuccinyl CH3), 1.68 (3H, s, CH3-30),
1.94 (3H, br s, AZT-CH3), 2.30–2.35, 2.40–2.51 (2H, m, AZT-H2-20),
2.35–2.42 (1H, m, H-19), 2.44, 2.49 (each 1H, d, J = 16.0 Hz, 28-O-
dimethylglutaryl H2-200), 2.49, 2.55 (each 1H, d, J = 16.0 Hz, 28-O-
dimethylglutaryl H2-400), 2.56, 2.68 (each 1H, d, J = 16.0 Hz, 3-O-
dimethylsuccinyl H2-20), 3.82, 4.26 (each 1H, d, J = 11.2 Hz, H2-
28), 4.07 (1H, q-like, AZT-H-40), 4.20–4.25 (1H, m, AZT-H-30), 4.26
(1H, dd, J = 4.0, 12.0 Hz, AZT-H-5a0), 4.43 (1H, dd, J = 4.4, 12.0 Hz,
AZT-H-5b0), 4.48 (1H, dd, J = 5.2, 10.8 Hz, H-3), 4.60, 4.69 (each
1H, br s, H2-29), 6.14 (1H, t-like, J = 6.4 Hz, AZT-H-10), 7.29 (1H,
d, J = 1.2 Hz, AZT-H-6), 9.43 (1H, br s, AZT-NH); 13C NMR (CDCl3,
100 MHz) d 12.5 AZT-CH3, 14.7 (C-27), 16.0 (C-26), 16.1 (C-25),
16.5 (C-24), 18.1 (C-6), 19.1 (C-30), 20.8 (C-11), 23.6 (C-2), 25.1
(C-12), 25.0, 25.6 (3-O-dimethylsuccinyl CH3), 27.0 (C-15), 27.9
(28-O-dimethylglutaryl CH3), 28.0 (C-23), 29.5 (C-21), 29.8 (C-
16), 32.6 (28-O-dimethylglutaryl C-300), 34.1 (C-7), 34.6 (C-22),
37.0 (C-10), 37.5 (AZT-C-20, C-13), 37.7 (C-4), 38.4 (C-1), 40.4 (3-
O-dimethylsuccinyl C-30), 40.8 (C-8), 42.7 (C-14), 44.5 (28-O-dim-
ethylglutaryl C-400), 44.7 (3-O-dimethylsuccinyl C-20), 45.0 (28-O-
dimethylglutaryl C-200), 46.2 (C-17), 47.7 (C-19), 48.7 (C-18), 50.2
(C-9), 55.4 (C-5), 60.4 (AZT-C-30), 62.7 (C-28), 62.8 (AZT-C-50),
81.5 (C-3), 81.8 (AZT-C-40), 85.3 (AZT-C-10), 109.9 (C-29), 111.2
(AZT-C-5), 135.4 (AZT-C-6), 150.0 (C-20), 150.1 (AZT-C-2), 163.9
(AZT-C-4), 171.0 (3-O-dimethylsuccinyl 10-COOꢂ), 171.2 (28-O-
dimethylglutaryl 500-COO-AZT), 172.3 (28-O-dimethylglutaryl 100-
COOꢂ), 182.1 (3-O-dimethylsuccinyl 40-COOY). HRESIMS (positive)
m/z 962.5881 [M+H]+ (calcd for C53H80N5O11, 962.5854).
Compound 55: ½a 3D0
ꢁ
ꢂ4.2° (c 2.99, CHCl3). 1H NMR (CDCl3,
400 MHz) d 0.76, 0.83 (each 3H, d, J = 8.0 Hz, CH3-29, 30), 0.82
(3H, s, CH3-24), 0.84 (6H, s, CH3-23, CH3-25), 0.94 (3H, s, CH3-
27), 1.01 (3H, s, CH3-26), 1.31 (6H, s, 3-O-dimethylsuccinyl CH3),
1.30, 1.31 (each 1H, s, 28-O-dimethylsuccinyl CH3), 1.93 (3H, br
s, AZT-CH3), 2.25–2.32, 2.44–2.50 (each 1H, m, AZT-H2-20), 2.57,
2.68 (each 1H, d, J = 15.6 Hz, 3-O-dimethylsuccinyl H2-20), 2.59,
2.73 (each 1H, d, J = 16.4 Hz, 28-O-dimethylsuccinyl H2-200), 3.81,
4.27 (each 1H, d, J = 10.8 Hz, H2-28), 4.09 (1H, q-like, AZT-H-40),
4.25–4.29 (1H, m, AZT-H-30), 4.28 (1H, dd, J = 4.4, 12.0 Hz, AZT-H-
5a0), 4.52 (1H, dd, J = 3.6, 12.0 Hz, AZT-H-5b0), 4.49 (1H, dd,
J = 5.2, 10.8 Hz, H-3), 6.16 (1H, t-like, J = 6.4 Hz, AZT-H-10), 7.27
(1H, br s, AZT-H-6), 9.31 (1H, br s, AZT-NH); 13C NMR (CDCl3,
100 MHz) d 12.4 AZT-CH3, 14.6 (C-27), 14.8 (C-29), 16.0 (C-26, C-
25), 16.5 (C-24), 18.1 (C-6), 20.8 (C-11), 21.6 (C-21), 22.8 (C-30),
23.6 (C-2), 25.1, 25.6 (3-O-dimethylsuccinyl CH3), 25.0, 25.8 (28-
O-dimethylsuccinyl CH3), 26.8 (C-12), 26.9 (C-15), 27.9 (C-23),
29.4 (C-20), 29.8 (C-16), 34.2 (C-7), 34.6 (C-22), 37.0 (C-10), 37.2
(C-13), 37.5 (AZT-C-20), 37.7 (C-4), 38.4 (C-1), 40.4 (3-O-dim-
ethylsuccinyl C-30), 40.7 (28-O-dimethylsuccinyl C-300), 40.9 (C-8),
42.8 (C-14), 44.5 (C-19), 44.6 (28-O-dimethylsuccinyl C-200), 44.7
(3-O-dimethylsuccinyl C-20), 46.5 (C-17), 48.1 (C-18), 49.9 (C-9),
55.4 (C-5), 60.4 (AZT-C-30), 63.1 (C-28), 63.3 (AZT-C-50), 81.5
(C-3), 82.0 (AZT-C-40), 84.9 (AZT-C-10), 111.3 (AZT-C-5), 135.2
(AZT-C-6), 150.2 (AZT-C-2), 164.0 (AZT-C-4), 171.0 (3-O-dim-
ethylsuccinyl 10-COOꢂ), 171.7 (28-O-dimethylsuccinyl 100-COOꢂ),
176.5 (28-O-dimethylsuccinyl 400-COO-AZT), 182.0 (3-O-dim-
ethylsuccinyl 40-COOY). HRESIMS (positive) m/z 950.5848 [M+H]+
(calcd for C52H80N5O11, 950.5854).
5.2.8.5. 30-Azido-30-deoxythymidine-50-yl 3-O-(30,30-dimethyl-
glutaryl)-lup-20(29)en-28-yl 3,3-dimethylglutarate (54). Yield
84.2% (starting from 107 mg of 40); white solid. ½a D26
ꢁ
+15.6° (c
3.09, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.84 (each 3H, s, CH3-24,
25), 0.85 (6H, s, CH3-23), 0.97 (3H, s, CH3-27), 1.02 (3H, s, CH3-26),
1.12, 1.15 (each 3H, s, 28-O-dimethylglutaryl CH3), 1.15 (6H, s,
3-O-dimethylglutaryl CH3), 1.68 (3H, s, CH3-30), 1.94 (3H, br s,
Compound 56: ½a 2D7
ꢁ
ꢂ2.5° (c 1.74, CHCl3). 1H NMR (CDCl3,
400 MHz) d 0.77, 0.84 (each 3H, d, J = 8.0 Hz, CH3-29, 30), 0.82
(3H, s, CH3-24), 0.85 (6H, s, CH3-23, CH3-25), 0.95 (3H, s, CH3-
27), 1.00 (3H, s, CH3-26), 1.31 (each 3H, s, 3-O-dimethylsuccinyl

J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
6467
CH3), 1.30, 1.33 (each 3H, s, 28-O-dimethylsuccinyl CH3), 1.95 (3H,
br s, AZT-CH3), 2.32–2.39, 2.44–2.50 (each 1H, m, AZT-H2-20), 2.57,
2.68 (each 1H, d, J = 15.6 Hz, 3-O-dimethylsuccinyl H2-20), 2.61,
2.66 (each 1H, d, J = 16.0 Hz, 28-O-dimethylsuccinyl H2-300), 3.81,
4.29 (each 1H, d, J = 10.8 Hz, H2-28), 4.04 (1H, q-like, AZT-H-40),
4.17–4.22 (1H, m, AZT-H-30), 4.26 (1H, dd, J = 3.6, 12.0 Hz, AZT-H-
5a0), 4.42 (1H, dd, J = 4.4, 12.0 Hz, AZT-H-5b0), 4.50 (1H, dd,
J = 5.2, 10.8 Hz, H-3), 6.13 (1H, t-like, J = 6.4 Hz, AZT-H-10), 7.30
(1H, d, br s, AZT-H-6), 9.16 (1H, br s, AZT-NH); 13C NMR (CDCl3,
100 MHz) d 12.5 AZT-CH3, 14.7 (C-27), 14.9 (C-29), 16.0 (C-26,
25), 16.5 (C-24), 18.2 (C-6), 20.8 (C-11), 21.6 (C-21), 22.9 (C-30),
23.6 (C-2), 25.1, 25.6 (3-O-dimethylsuccinyl CH3), 25.5, 25.8 (28-
O-dimethylsuccinyl CH3), 26.8 (C-12), 26.9 (C-15), 27.9 (C-23),
29.4 (C-20), 30.0 (C-16), 34.2 (C-7), 34.7 (C-22), 37.0 (C-10), 37.2
(C-13), 37.5 (AZT-C-20), 37.7 (C-4), 38.4 (C-1), 40.4 (3-O-dim-
ethylsuccinyl C-30), 40.9 (C-8), 41.1 (28-O-dimethylsuccinyl C-200),
42.9 (C-14), 43.9 (28-O-dimethylsuccinyl C-300), 44.6 (C-19), 44.7
(3-O-dimethylsuccinyl C-20), 46.7 (C-17), 48.2 (C-18), 50.0 (C-9),
55.4 (C-5), 60.3 (AZT-C-30), 63.0 (AZT-C-50), 63.4 (C-28), 81.5 (C-
3), 81.8 (AZT-C-40), 85.3 (AZT-C-10), 111.3 (AZT-C-5), 135.4 (AZT-
C-6), 150.0 (AZT-C-2), 163.8 (AZT-C-4), 170.8 (28-O-dimethylsucci-
nyl 400-COOꢂFZN), 171.0 (3-O-dimethylsuccinyl 10-COOꢂ), 176.9
(28-O-dimethylsuccinyl 100-COO–), 181.7 (3-O-dimethylsuccinyl
40-COOY). HRESIMS (positive) m/z 950.5873 [M+H]+ (calcd for
J = 7.2 Hz, 3-O-glutaryl H2-30), 2.26–2.35, 2.44–2.51 (each 1H, m,
AZT-H2-20), 2.40 (2H, t, J = 7.2 Hz, 3-O-glutaryl H2-20), 2.44 (2H, t,
J = 7.2 Hz, 3-O-glutaryl H2-40), 2.60, 2.74 (each 1H, d, J = 16.0 Hz,
28-O-dimethylsuccinyl H2-200), 3.82, 4.28 (each 1H, d, J = 10.8 Hz,
H2-28), 4.09 (1H, q-like, AZT-H-40), 4.25–4.30 (1H, m, AZT-H-30),
4.28 (1H, dd, J = 4.0, 12.0 Hz, AZT-H-5a0), 4.51 (1H, dd, J = 4.0,
12.0 Hz, AZT-H2-5b0), 4.49 (1H, dd, J = 5.2, 10.0 Hz, H-3), 6.16 (1H,
t-like, J = 6.4 Hz, AZT-H-10), 7.28 (1H, d, J = 1.2 Hz, AZT-H-6), 8.96
(1H, br s, AZT-NH); 13C NMR (CDCl3, 100 MHz) d 12.4 AZT-CH3,
14.6 (C-27), 14.9 (C-29), 16.0 (C-26), 16.1 (C-25), 16.6 (C-24),
18.2 (C-6), 20.1 (3-O-glutaryl C-30), 20.8 (C-11), 21.6 (C-21), 22.9
(C-30), 23.7 (C-2), 25.1, 25.9 (28-O-dimethylsuccinyl CH3), 26.8
(C-12), 26.9 (C-15), 28.0 (C-23), 29.4 (C-20), 29.9 (C-16), 32.9 (3-
O-glutaryl C-40), 33.7 (3-O-glutaryl C-20), 34.2 (C-7), 34.7 (C-22),
37.1 (C-10), 37.2 (C-13), 37.5 (AZT-C-20), 37.8 (C-4), 38.4 (C-1),
40.7 (28-O-dimethylsuccinyl C-300), 40.9 (C-8), 42.9 (C-14), 44.6
(C-19, 28-O-dimethylsuccinyl C-200), 46.5 (C-17), 48.2 (C-18), 50.0
(C-9), 55.4 (C-5), 60.4 (AZT-C-30), 63.2 (C-28), 63.3 (AZT-C-50),
81.1 (C-3), 82.0 (AZT-C-40), 84.9 (AZT-C-10), 111.3 (AZT-C-5),
135.2 (AZT-C-6), 150.0 (AZT-C-2), 163.7 (AZT-C-4), 171.7 (28-O-
dimethylsuccinyl 100-COOꢂ), 172.7 (3-O-glutaryl 10-COOꢂ), 176.5
(28-O-dimethylsuccinyl 400-COO-AZT), 177.1 (3-O-glutaryl 50-
COOY). HRESIMS (positive) m/z 936.5716 [M+H]+ (calcd for
C51H78N5O11, 936.5698).
C
52H80N5O11, 950.5854).
5.2.8.9.
4-(30-Azido-30-deoxythymidine-50-yl)-1-(3-O-glutaryl-
5.2.8.7. 30-Azido-30-deoxythymidine-50-yl 3-O-(30,30-dimethyl-
succinyl)-lup-28-yl 3,3-dimethylglutarate (57). Yield 45.6%
lup-28-yl) 2,2-dimethylsuccinate (59). Yield 66.1% (starting from
32.6 mg of 45); white solid. ½a D27
ꢁ
+2.3° (c 1.66, CHCl3). 1H NMR
(starting from 84 mg of 43); white solid. ½a D27
ꢁ
ꢂ0.02° (c 2.12, CHCl3).
(CDCl3, 400 MHz) d 0.77, 0.84 (each 3H, d, J = 8.0 Hz, CH3-29, 30),
0.85 (6H, s, CH3-24, CH3-23), 0.87 (3H, s, CH3-25), 0.96 (3H, s,
CH3-27), 1.04 (3H, s, CH3-26), 1.31, 1.34 (each 3H, s, 28-O-dim-
ethylsuccinyl CH3), 1.96 (3H, br s, AZT-CH3), 1.98 (2H, quint,
J = 7.2 Hz, 3-O-glutaryl H2-30), 2.32–2.51 (2H, m, AZT-H2-20), 2.40
(2H, t, J = 7.2 Hz, 3-O-glutaryl H2-20), 2.43 (2H, t, J = 7.2 Hz, 3-O-
glutaryl H2-40), 2.61, 2.66 (each 1H, d, J = 16.4 Hz, 28-O-dim-
ethylsuccinyl H2-200), 3.81, 4.30 (each 1H, d, J = 10.8 Hz, H2-28),
4.05 (1H, q-like, AZT-H-40), 4.17–4.23 (1H, m, AZT-H-30), 4.26
(1H, dd, J = 3.6, 12.4 Hz, AZT-H-5a0), 4.42 (1H, dd, J = 4.4, 12.4 Hz,
AZT-H-5b0), 4.49 (1H, dd, J = 5.2, 10.8 Hz, H-3), 6.14 (1H, t-like,
J = 6.4 Hz, AZT-H-10), 7.31 (1H, d, J = 0.8 Hz, AZT-H-6), 9.14 (1H,
br s, AZT-NH); 13C NMR (CDCl3, 100 MHz) d 12.5 AZT-CH3, 14.7
(C-27), 14.9 (C-29), 16.0 (C-26), 16.1 (C-25), 16.6 (C-24), 18.2 (C-
6), 20.1 (3-O-glutaryl C-30), 20.8 (C-11), 21.6 (C-21), 22.9 (C-30),
23.7 (C-2), 25.5, 25.8 (28-O-dimethylsuccinyl CH3), 26.8 (C-12),
26.9 (C-15), 28.0 (C-23), 29.4 (C-20), 29.9 (C-16), 32.9 (3-O-glutaryl
C-40), 33.7 (3-O-glutaryl C-20), 34.2 (C-7), 34.7 (C-22), 37.0 (C-10),
37.2 (C-13), 37.5 (AZT-C-20), 37.8 (C-4), 38.4 (C-1), 40.9 (C-8),
41.1 (28-O-dimethylsuccinyl C-200), 42.9 (C-14), 43.9 (28-O-dim-
ethylsuccinyl C-300), 44.6 (C-19), 46.7 (C-17), 48.2 (C-18), 50.0 (C-
9), 55.4 (C-5), 60.3 (AZT-C-30), 62.9 (AZT-C-50), 63.3 (C-28), 81.1
(C-3), 81.8 (AZT-C-40), 85.2 (AZT-C-10), 111.3 (AZT-C-5), 135.5
(AZT-C-6), 150.1 (AZT-C-2), 163.8 (AZT-C-4), 170.8 (28-O-dim-
ethylsuccinyl 400-COO-AZT), 172.7 (3-O-glutaryl 10-COOꢂ), 176.9
(28-O-dimethylsuccinyl 100-COOꢂ), 177.3 (3-O-glutaryl 50-COOY).
1H NMR (CDCl3, 400 MHz) d 0.77, 0.84 (each 3H, d, J = 8.0 Hz, CH3-
29, 30), 0.82 (3H, s, CH3-24), 0.85 (6H, s, CH3-23, CH3-25), 0.95 (3H,
s, CH3-27), 1.03 (3H, s, CH3-26), 1.12, 1.15 (each 3H, s, 28-O-dim-
ethylglutaryl CH3), 1.30, 1.31 (each 3H, s, 3-O-dimethylsuccinyl
CH3), 1.95 (3H, br s, AZT-CH3), 2.33–2.40, 2.43–2.50 (2H, m, AZT-
H2-20), 2.42, 2.48 (each 1H, d, J = 14.4 Hz, 28-O-dimethylglutaryl
H2-200), 2.49, 2.56 (each 1H, d, J = 15.2 Hz, 28-O-dimethylglutaryl
H2-400), 2.57, 2.68 (each 1H, d, J = 15.6 Hz, 3-O-dimethylsuccinyl
H2-20), 3.79, 4.26 (each 1H, d, J = 10.8 Hz, H2-28), 4.07 (1H, q-like,
AZT-H-40), 4.20–4.23 (1H, m, AZT-H-30), 4.26 (1H, dd, J = 3.6,
12.0 Hz, AZT-H-5a0), 4.44 (1H, dd, J = 4.4, 12.0 Hz, AZT-H-5b0),
4.50 (1H, dd, J = 5.2, 10.4 Hz, H-3), 6.14 (1H, t-like, J = 6.4 Hz,
AZT-H-10), 7.29 (1H, br s, AZT-H-6), 9.15 (1H, br s, AZT-NH); 13C
NMR (CDCl3, 100 MHz) d 12.5 AZT-CH3, 14.6 (C-27), 14.9 (C-29),
16.0 (C-26, 25), 16.5 (C-24), 18.2 (C-6), 20.8 (C-11), 21.6 (C-21),
22.9 (C-30), 23.6 (C-2), 25.1, 25.6 (3-O-dimethylsuccinyl CH3),
26.8 (C-12), 26.9 (C-15), 27.9 (28-O-dimethylglutaryl CH3), 28.0
(C-23), 29.4 (C-20), 29.9 (C-16), 32.6 (28-O-dimethylglutaryl
C-300), 34.2 (C-7), 34.8 (C-22), 37.0 (C-10), 37.2 (C-13), 37.6 (AZT-
C-20), 37.7 (C-4), 38.4 (C-1), 40.4 (3-O-dimethylsuccinyl C-30),
40.9 (C-8), 42.9 (C-14), 44.5 (28-O-dimethylglutaryl C-400, C-19),
44.7 (3-O-dimethylsuccinyl C-20), 45.1 (28-O-dimethylsuccinyl C-
200), 46.4 (C-17), 48.1 (C-18), 50.0 (C-9), 55.4 (C-5), 60.5
(AZT-C-30), 62.8 (C-28, AZT-C-50), 81.5 (C-3), 81.9 (AZT-C-40), 85.3
(AZT-C-10), 111.3 (AZT-C-5), 135.4 (AZT-C-6), 150.1 (AZT-C-2),
163.8 (AZT-C-4), 171.0 (3-O-dimethylsuccinyl 10-COOꢂ), 171.2
(28-O-dimethylglutaryl 500-COO-AZT), 172.3 (28-O-dimethylgluta-
ryl 100-COOꢂ), 181.8 (3-O-dimethylsuccinyl 40-COOY). HRESIMS
(positive) m/z 964.6037 [M+H]+ (calcd for C53H82N5O11, 964.6011).
HRESIMS (positive) m/z 936.5694 [M+H]+ (calcd for C51H78N5O11
936.5698).
,
5.2.8.10. 30-Azido-30-deoxythymidine-50-yl 3-O-glutaryl-lup-28-
yl glutarate (60). Yield 58.7% (starting from 41.2 mg of 46); white
5.2.8.8.
1-(30-Azido-30-deoxythymidine-50-yl)-4-(3-O-glutaryl-
solid. ½a 2D7
ꢁ
+7.0° (c 1.40, CHCl3). 1H NMR (CDCl3, 400 MHz) d 0.78,
lup-28-yl) 2,2-dimethylsuccinate (58). Yield 82.5% (starting from
0.84 (each 3H, d, J = 8.0 Hz, CH3-29, 30), 0.84 (6H, s, CH3-24, CH3-
23), 0.87 (3H, s, CH3-25), 0.96 (3H, s, CH3-27), 1.05 (3H, s, CH3-
26), 1.94 (3H, d, J = 1.2 Hz, AZT-CH3), 1.97 (2H, quint, J = 7.2 Hz,
3-O-glutaryl H2-30), 1.99 (2H, quint, J = 7.2 Hz, 28-O-glutaryl H2-
300), 2.34–2.51 (2H, m, AZT-H2-20), 2.40 (2H, t, J = 7.2 Hz, 3-O-glut-
aryl H2-20), 2.41 (2H, t, J = 7.2 Hz, 28-O-glutaryl H2-200), 2.43 (2H,
20.4 mg of 44); white solid. ½a D27
ꢁ
ꢂ1.9° (c 1.11, CHCl3). 1H NMR
(CDCl3, 400 MHz) d 0.77, 0.84 (each 3H, d, J = 8.0 Hz, CH3-29, 30),
0.85 (6H, s, CH3-24, CH3-23), 0.87 (3H, s, CH3-25), 0.95 (3H, s,
CH3-27), 1.02 (3H, s, CH3-26), 1.30, 1.31 (each 3H, s, 28-O-dim-
ethylsuccinyl CH3), 1.94 (3H, br s, AZT-CH3), 1.98 (2H, quint,

6468
J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
t, J = 7.2 Hz, 3-O-glutaryl H2-40), 2.46 (2H, t, J = 7.2 Hz, 28-O-gluta-
ryl H2-400), 3.83, 4.29 (each 1H, d, J = 11.2 Hz, H2-28), 4.07 (1H, q-
like, AZT-H-40), 4.20–4.24 (1H, m, AZT-H-30), 4.32 (1H, dd, J = 4.0,
12.0 Hz, AZT-H-5a0), 4.40 (1H, dd, J = 4.4, 12.0 Hz, AZT-H-5b0),
4.49 (1H, dd, J = 6.0, 10.4 Hz, H-3), 6.09 (1H, t-like, J = 6.4 Hz,
AZT-H-10), 7.23 (1H, br s, AZT-H-6), 9.10 (1H, br s, AZT-NH); 13C
NMR (CDCl3, 100 MHz) d 12.5 AZT-CH3, 14.6 (C-27), 14.9 (C-29),
16.0 (C-26, C-25), 16.6 (C-24), 18.2 (C-6), 20.1 (28-O-glutaryl C-
300, 3-O-glutaryl C-30), 20.8 (C-11), 21.6 (C-21), 22.9 (C-30), 23.7
(C-2), 26.8 (C-12), 26.9 (C-15), 28.0 (C-23), 29.4 (C-20), 29.9 (C-
16), 32.9 (3-O-glutaryl C-40), 33.1 (28-O-glutaryl C-400), 33.2 (28-
O-glutaryl C-200), 33.7 (3-O-glutaryl C-20), 34.2 (C-7), 34.7 (C-22),
37.0 (C-10), 37.2 (C-13), 37.5 (AZT-C-20), 37.8 (C-4), 38.4 (C-1),
41.0 (C-8), 42.9 (C-14), 44.6 (C-19), 46.6 (C-17), 48.2 (C-18), 50.0
(C-9), 55.4 (C-5), 60.6 (AZT-C-30), 63.0 (C-28), 63.3 (AZT-C-50),
81.1 (C-3), 81.8 (AZT-C-40), 85.7 (AZT-C-10), 111.3 (AZT-C-5),
135.5 (AZT-C-6), 150.0 (AZT-C-2), 163.7 (AZT-C-4), 172.3 (28-O-
glutaryl 500-COO-AZT), 172.7 (3-O-glutaryl 10-COOꢂ), 173.1 (28-O-
glutaryl 100-COOꢂ), 177.2 (3-O-glutaryl 50-COOY). HRESIMS (posi-
tive) m/z 922.5557 [M+H]+ (calcd for C60H76N5O11, 922.5541).
1H, d, J = 10.8 Hz, H2-28), 4.07 (1H, q-like, AZT-H-40), 4.20–4.28
(1H, m, AZT-H-30), 4.26 (1H, dd, J = 3.6, 12.0 Hz, AZT-H-5a0), 4.44
(1H, dd, J = 4.4, 12.0 Hz, AZT-H-5b0), 4.50 (1H, dd, J = 5.2, 10.8 Hz,
H-3), 6.14 (1H, t-like, J = 6.4 Hz, AZT-H-10), 7.29 (1H, d, J = 1.2 Hz,
AZT-H-6), 9.19 (1H, br s, AZT-NH); 13C NMR (CDCl3, 100 MHz) d
12.5 AZT-CH3, 14.6 (C-27), 14.9 (C-29), 16.0 (C-26, C-25), 16.6 (C-
24), 18.2 (C-6), 20.8 (C-11), 21.6 (C-21), 22.9 (C-30), 23.8 (C-2),
26.8 (C-12), 26.9 (C-15), 27.9 (28-O-dimethylglutaryl CH3), 27.9,
28.0 (3-O-dimethylglutaryl CH3), 28.0 (C-23), 29.4 (C-20), 29.9
(C-16), 32.6 (28-O-dimethylglutaryl C-300), 32.7 (3-O-dimethylglut-
aryl C-30), 34.2 (C-7), 34.7 (C-22), 37.0 (C-10), 37.2 (C-13), 37.6
(AZT-C-20), 37.7 (C-4), 38.4 (C-1), 40.9 (C-8), 42.9 (C-14), 44.6
(28-O-dimethylglutaryl C-400 and C-19), 45.1 (28-O-dimethylgluta-
ryl C-200), 45.2 (3-O-glutaryl C-40), 45.7 (3-O-glutaryl C-20), 46.4 (C-
17), 48.1 (C-18), 50.0 (C-9), 55.4 (C-5), 60.4 (AZT-C-30), 62.7 (C-28,
and AZT-C-50), 81.4 (C-3), 81.8 (AZT-C-40), 85.3 (AZT-C-10), 111.3
(AZT-C-5), 135.4 (AZT-C-6), 150.1 (AZT-C-2), 163.8 (AZT-C-4),
171.2 (28-O-dimethylglutaryl 500-COO-AZT), 172.3 (28-O-dimeth-
ylglutaryl 100-COOꢂ, 3-O-dimethylglutaryl 10-COOꢂ), 175.6 (3-O-
dimethylglutaryl 50-COOY). HRESIMS (positive) m/z 1000.5991
[M+Na]+ (calcd for C54H83N5O11Na, 1000.5987).
5.2.8.11. 30-Azido-30-deoxythymidine-50-yl 3-O-glutaryl-lup-28-
yl 3,3-dimethylglutarate (61). Yield 75.6% (starting from 39 mg of
5.3. HIV-1NL4-3 inhibition assay in MT-4 lymphocytes
47); white solid. ½a D27
ꢁ
+3.5° (c 2.13, CHCl3). 1H NMR (CDCl3,
400 MHz) d 0.77, 0.84 (each 3H, d, J = 8.0 Hz, CH3-29, 30), 0.85
(6H, s, CH3-24, CH3-23), 0.86 (3H, s, CH3-25), 0.95 (3H, s, CH3-
27), 1.03 (3H, s, CH3-26), 1.12, 1.15 (each 3H, s, 28-O-dimethylglut-
aryl CH3), 1.95 (3H, d, J = 1.2 Hz, AZT-CH3), 1.97 (2H, quint,
J = 7.2 Hz, 3-O-glutaryl H2-30), 2.32–2.51 (2H, m, AZT-H2-20), 2.40
(2H, t, J = 7.2 Hz, 3-O-glutaryl H2-20), 2.42, 2.49 (each 1H, d,
J = 14.4 Hz, 28-O-dimethylglutaryl H2-200), 2.43 (2H, t, J = 7.2 Hz,
3-O-glutaryl H2-40), 2.48, 2.56 (each 1H, d, J = 15.2 Hz, 28-O-dim-
ethylglutaryl H2-400), 3.79, 4.27 (each 1H, d, J = 10.8 Hz, H2-28),
4.07 (1H, q-like, AZT-H-40), 4.20–4.28 (1H, m, AZT-H-30), 4.26
(1H, dd, J = 3.6, 12.4 Hz, AZT-H-5a0), 4.44 (1H, dd, J = 4.4, 12.4 Hz,
AZT-H-5b0), 4.49 (1H, dd, J = 5.2, 10.8 Hz, H-3), 6.14 (1H, t-like,
J = 6.4 Hz, AZT-H-10), 7.30 (1H, d, J = 0.8 Hz, AZT-H-6), 9.18 (1H,
br s, AZT-NH); 13C NMR (CDCl3, 100 MHz) d 12.5 AZT-CH3, 14.6
(C-27), 14.9 (C-29), 16.0 (C-26, C-25), 16.6 (C-24), 18.2 (C-6), 20.1
(3-O-glutaryl C-30), 20.8 (C-11), 21.6 (C-21), 22.9 (C-30), 23.7 (C-
2), 26.8 (C-12), 26.9 (C-15), 27.9, 28.0 (28-O-dimethylglutaryl
CH3), 28.0 (C-23), 29.4 (C-20), 29.9 (C-16), 32.6 (28-O-dimethyl-
glutaryl C-300), 32.9 (3-O-glutaryl C-40), 33.7 (3-O-glutaryl C-20),
34.2 (C-7), 34.7 (C-22), 37.0 (C-10), 37.2 (C-13), 37.6 (AZT-C-20),
37.8 (C-4), 38.4 (C-1), 40.9 (C-8), 42.9 (C-14), 44.5 (28-O-dimethyl-
glutaryl C-400 and C-19), 45.1 (28-O-dimethylglutaryl C-200), 46.4
(C-17), 48.1 (C-18), 50.0 (C-9), 55.4 (C-5), 60.4 (AZT-C-30), 62.7
(C-28 and AZT-C-50), 81.1 (C-3), 81.9 (AZT-C-40), 85.3 (AZT-C-10),
111.3 (AZT-C-5), 135.4 (AZT-C-6), 150.1 (AZT-C-2), 163.8 (AZT-C-
4), 171.2 (28-O-dimethylglutaryl 500-COO-AZT), 172.3 (28-O-dim-
ethylglutaryl 100-COOꢂ), 172.7 (3-O-glutaryl 10-COOꢂ), 177.3 (3-
O-glutaryl 50-COOY). HRESIMS (positive) m/z 950.5876 [M+H]+
(calcd for C52H80N5O11, 950.5854).
A
previously described procedure was used in the
experiments.21,23
Acknowledgments
This investigation was supported by Grant-in-Aid for Scientific
Research (C) from the Japan Society for the Promotion of Science
(Y.K.) as well as a Grant AI-077417 from the National Institute of
Allergy and Infectious Diseases (K.-H.L.).
Supplementary data
Supplementary data associated with this article can be found, in
the online version, at doi:10.1016/j.bmc.2010.06.092. These data
include MOL files and InChiKeys of the most important compounds
described in this article.
References and notes
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H2-20), 2.41, 2.47 (each 1H, d, J = 14.4 Hz, 3-O-dimethylglutaryl
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H2-200), 2.47 (2H, s, 3-O-dimethylglutaryl H2-40), 2.48, 2.56 (each
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6
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M, TI 21,515; 6: EC50
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l
2 ꢀ 10ꢂ5
l
M, TI 1.12 ꢀ 106; 5: EC50 0.00066
l
0.00087 lM, TI 42,400.
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J. Xiong et al. / Bioorg. Med. Chem. 18 (2010) 6451–6469
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