Cytotoxicity of Rhamnosylanthraquinones
J ournal of Natural Products, 2001, Vol. 64, No. 9 1167
2
3
77 (4.39), 225 (4.68), 203 (4.85) nm; IR (CHCl
3
) νmax 3681,
432 (3.96), 299 (3.92), 288 (4.06), 261 (4.24), 225 (4.48) nm; IR
625 (OH), 1630 (CO) cm-1; 1H and C NMR, Tables 2 and 3;
13
(CHCl
(400 MHz, CDCl
) νmax 3690 (OH), 1755, 1628, 1609 (CO) cm ; H NMR
-1 1
3
FABMS m/z 747 (M + Li) (70), 492 (8), 313 (37), 256 (40), 160
3
) δ 12.5 (1H, s, OH-8), 12.2 (1H, s, OH-1),
+
(
7
100); HRCIMS [M + H] m/z 741.2171 (calcd for C40
41.2183).
P r in oid in -em od in bia n th r on e (9B): amorphous powder;
H
37
O
14
6.68 (1H, brs, H-2), 6.70 (2H, brs, H-4, H-7), 6.55 (1H, brs,
H-5), 5.55 (1H, brs, H-1′), 5.44 (1H, t, J ) 3.8 Hz, H-2′), 5.40
(1H, m, H-3′), 5.17 (1H, t, J ) 9.7 Hz, H-4′), 5.05 (1H, br s,
R]2
5
+6° (c 0.76, CHCl
); UV (EtOH) λmax (log ꢀ) 363 (4.46),
) νmax 3681,
625 (OH), 1630 (CO) cm-1; 1H and C NMR, Tables 2 and 3;
[
2
3
D
3
H-10), 3.90 (1H, m, H-5′), 2.35 (3H, s, CH
CO-2′′), 2.02 (6H, s, CH CO-3′′, CH CO-4′′), 1.82 (3H, s, CH
11), 1.25 (3H, d, J ) 6 Hz, CH
3
-3), 2.22 (3H, s, CH
3
3
3
-
-
)
77 (4.39), 225 (4.68), 203 (4.85) nm; IR (CHCl
3
3
3
13
13
3
-6′); C NMR (75 MHz, CDCl
FABMS m/z 747 (M + Li) (70), 492 (8), 313 (37), 256 (40), 160
δ 202.0 (C-11), 191.6 (C-9), 171.2 (COC-3′), 170.0 (COC-2′,
COC-4′), 165.6 (C-8), 163.3 (C-1), 161.9 (C-6), 148.7 (C-3), 140.1
(C-4a), 137.5 (C-5a), 120.1 (C-4), 117.9 (C-2), 112.5 (C-1a),
110.5 (C-8a), 108.1 (C-5), 103.9 (C-7), 95.4 (C-1′), 70.6 (C-2′),
+
(
7
100); HRCIMS [M + H] m/z 741.2171 (calcd for C40
41.2183).
P r in oid in -em od in bia n th r on e (9C): amorphous powder;
37 14
H O
R]2
5
0° (c 0.6, CHCl
); H and C NMR, Tables 2 and 3;
1
13
[
D
3
69.5 (C-3′), 69.0 (C-4′), 68.2 (C-5′), 59.1 (C-10), 22.2 (CH
20.9 (CH CO-3′′), 20.8 (CH CO-2′′, CH CO-4′′), 17.5 (CH
EIMS m/z 542 [M] (25), 498 (15), 273 (100), 241 (80); HRCIMS
3
-3),
FABMS m/z 747 (M + Li) (70), 492 (8), 313 (37), 256 (40), 160
3
3
3
3
-6′);
+
+
(
7
100); HRCIMS [M + H] m/z 741.2171 (calcd for C40
41.2183).
P r in oid in -em od in bia n th r on e (9D): amorphous powder;
37 14
H O
+
m/z 543.1493 [M + H] (calcd for C27
P r ep a r a tion of Com p ou n d 13. A solution of prinoidin (1)
20 mg, 0.04 mmol) in (Ac) O (4 mL) and pyridine (25 µL) was
stirred for 5 h at room temperature. After washing, the
reaction mixture was dried (Na SO ), and the solvent evapo-
rated. The residue, after preparative TLC, gave compound 12
27
H O12 543.1503).
R]2
5
1
13
[
D
3
-16.5° (c 0.4 CHCl ); H and C NMR, Tables 2 and 3;
(
2
FABMS m/z 747 (M + Li) (70), 492 (8), 313 (37), 256 (40), 160
+
(
7
37 14
100); HRCIMS [M + H] m/z 741.2171 (calcd for C40H O
2
4
41.2183).
P r in oid in bia n th r on e (10A): yellow crystals; mp 144-
(
17 mg, yield 76%) as an amorphous powder: UV (MeOH) λmax
2
5
1
3
3
47 °C; [R]
D
+133° (c 1.06, CHCl
3
); UV (MeOH) λmax (log ꢀ)
) νmax 3677,
(log ꢀ) 357 (3.86), 271 (4.06), 221 (4.12) nm; IR (CHCl
3
) νmax
61 (4.56), 277 (4.49), 209 (4.89) nm; IR (CHCl
505 (OH), 1748 (ester), 1637, 1619, 1607 (CO) cm ; H and
3
-1
1
3
693 (OH), 1755, 1628, 1609 (CO) cm ; H NMR (400 MHz,
) δ 12.2 (1H, s, OH-8), 12.0 (1H, s, OH-1), 7.58 (1H, d, J
2 Hz, H-4), 7.45 (1H, d, J ) 2 Hz, H-5), 7.05 (1H, d, J ) 2
-
1 1
CDCl
)
3
1
3
+
C NMR, Tables 2 and 3; CIMS m/z 993 [M + Na] (100), 971
+
+
[
M + 1] , 508 (50), 360 (25), 279 (27), 150 (70); HRCIMS [MH]
Hz, H-2), 6.87 (1H, d, J ) 2 Hz, H-7), 5.57 (1H, d, J ) 2 Hz,
H-1′), 5.40 (2H, m, H-2′, H-3′), 5.10 (1H, t, J ) 9.5 Hz, H-4′),
m/z 971.2975 (calcd for C50
51
H O20 971.2974).
P r in oid in bia n th r on e (10B): yellow crystals; mp 155-
5
(
.05 (1H, s, H-10), 3.85 (1H, m, H-5′), 2.37 (3H, s, CH
3H, s, CH CO-2′), 2.00 (6H, s, CH CO-3′, CH
), 1.15 (3H, d, J ) 6 Hz, CH
), 191.2 (C-9), 181.7 (C-10), 170.5 (COC-
′), 170.1 (COC-3′, COC-4′), 164.9 (C-8), 162.7 (C-1), 162.2 (C-
), 148.9 (C-3), 135.5 (C-5a), 133.1 (C-4a), 124.7 (C-2), 121.6
3
-3), 2.05
2
5
1
3
3
57 °C; [R]
D
+127° (c 0.96, CHCl
3
); UV (MeOH) λmax (log ꢀ)
) νmax 3677,
3
3
3
CO-4′), 1.82 (3H,
3
-6′); C NMR (75 MHz,
62 (4.44), 277 (4.39), 210 (4.75) nm; IR (CHCl
475 (OH), 1748 (ester), 1637, 1619, 1605 (CO) cm ; H and
3
13
s, COCH
CDCl
2
6
3
-
1 1
3
) δ 202 (CHOCH
3
1
3
+
C NMR, Tables 2 and 3; CIMS m/z 993 [M + Na] (100), 971
+
+
[
M + 1] , 508 (50), 360 (25), 279 (27), 150 (70); HRCIMS [MH]
m/z 971.2975 (calcd for C50
51
H O20 971.2974).
(
(
(
C-4), 113.6 (C-1a), 111.7 (C-8a), 109.6 (C-5), 109.3 (C-7), 95.4
C-1′), 70.6 (C-2′), 69.2 (C-3′), 69.7 (C-4′), 68.0 (C-5′), 24.7
Rh a m n ep a lin (11A): yellow amorphous powder; mp 177-
2
5
1
3
3
79 °C; [R]
D
+47.2° (c 0.7, CHCl
3
); UV (MeOH) λmax (log ꢀ)
) νmax 3683,
CHOCH
CO-4′), 17.5 (CH
98 (10), 273 (15), 256 (100); HRCIMS m/z 571.1805 [M + H]
calcd for C29 12 571.1816).
Dim er iza tion of Em od in An th r on e a n d Ch r ysop h a n ol
3
), 22.3 (CH
3
-3), 20.9 (CH
-6′); EIMS m/z 570 [M] (35), 528 (55),
3 3
CO-3′), 20.8 (CH CO-2′,
61 (4.46), 278 (4.32), 206 (4.82) nm; IR (CHCl
3
+
CH
2
(
3
3
-
1 1
657 (OH), 1747 (ester), 1636, 1619, 1605 (CO) cm ; H and
C NMR, Tables 2 and 3; CIMS m/z 969 [M - 1] (100), 682+
10), 516 (20), 485 (35); HRFABMS m/z 971.2979 [M + H]
+
1
3
31
H O
(
(
calcd for C50
51
H O20 971.2972).
An th r on e: Com p ou n d s 5A,B, 6A-D, a n d 13A,B. A solution
of emodin (108 mg, 0.4 mmol) or chrysophanol (108 mg, 0.4
Rh a m n ep a lin (11B): yellow amorphous powder; mp 153-
2
5
1
3
3
55 °C; [R]
D
+ 69° (c 0.96, CHCl
3
); UV (MeOH) λmax (log ꢀ)
) νmax 3680,
mmol) in acetic acid (10 mL) was added to a solution of SnCl
303 mg) in concentrated HCl (0.9 mL), and the reaction
mixture was stirred for 5 h at 80 °C. After the addition of
water, the reaction mixture was extracted by CH Cl . The
organic phases were dried (Na SO ) and evaporated to yield
emodin anthrone (79 mg, yield 74%) or chrysophanol anthrone
98 mg, yield 94%). To a solution of emodin anthrone (72 mg)
and chrysophanol anthrone (77 mg) in EtOH (35 mL) was
added a solution of FeCl (0.2 g) in EtOH (21 mL). The reaction
mixture was stirred for 3 h under reflux, then, after addition
of a solution (1 L) of 5% HCl, extracted by CH Cl . The
combined organic phases were washed, dried (Na SO ), and
2
60 (4.42), 277 (4.37), 207 (4.79) nm; IR (CHCl
3
(
-
1 1
657 (OH), 1747 (ester), 1636, 1619, 1605 (CO) cm ; H and
C NMR, Tables 2 and 3; CIMS m/z 969 [M - 1] (100), 682+
10), 516 (20), 485 (35); HRFABMS m/z 971.2979 [M + H]
1
3
2
2
(
(
2
4
calcd for C50
51
H O20 971.2972).
Rh a m n ep a lin (11C): yellow amorphous powder; mp 167-
(
2
5
1
3
3
70 °C; [R]
D
+56.2° (c 1.12, CHCl
3
); UV (MeOH) λmax (log ꢀ)
) νmax 3680,
61 (4.42), 277 (4.36), 206 (4.78) nm; IR (CHCl
3
-
1 1
3
657 (OH), 1747 (ester), 1636, 1619, 1604 (CO) cm ; H and
C NMR, Tables 2 and 3; CIMS m/z 969 [M - 1] (100), 682+
10), 516 (20), 485 (35); HRFABMS m/z 971.2979 [M + H]
1
3
2
2
(
(
2
4
calcd for C50
Acetyla tion of Com p ou n d 2. A solution of 2′,3′-di-O-
acetylfrangulin A (2) (10 mg, 0.02 mmol) in (Ac) O (1 mL) and
pyridine (1 mL) was stirred for 24 h at room temperature. After
51
H O20 971.2972).
evaporated. The residue, after preparative TLC and HPLC of
the isolated fractions on a chiral OD column (heptane-
2
2
5
-propanol-acetic acid 8:2:0.02), gave compounds 5A (4.2 mg),
B (3 mg), 6A (6 mg), 6B (5.8 mg), 6C (8.1 mg), 6D (7 mg), 7A
addition of water, the reaction mixture was extracted by CH
Cl . The combined organic phases were washed, dried (Na
SO
compound 4 (10.2 mg, yield 81%) as an amorphous powder:
2
-
(8.2 mg), and 7B (10 mg).
2
2
-
In Vivo Bioa ssa y of P r in oid in (1). Prinoidin (1) was
4
), and evaporated. The residue, after preparative TLC, gave
injected intraperitoneally to two mice CDF
P388 leukaemia cells according to a published technique.
1
grafted i.v. with
1
8
2
5
[
R]
literature values.
P r ep a r a tion of Com p ou n d 12. To a solution of 2′,3′-di-
O-acetylfrangulin A (2) (10 mg, 0.02 mmol) in (Ac) O (3 mL)
and CH Cl (3 mL) was added 4-DMAP (3 mL), and the
D
-80.8° (c 0.5, CHCl
3
), and other data comparable with
KB Cytotoxicity Assa y. The assays were performed ac-
cording to a published technique.19 The control used for
comparison was doxorubicin (IC50 0.058 µg/mL).
2
2
2
reaction mixture was stirred for 30 min at room temperature.
After the addition of water, the reaction mixture was extracted
Ack n ow led gm en t. This study has been carried out in the
framework of a program of cooperation between CNRS (France)
and NCST (Vietnam). The authors acknowledge Mrs. M.-T.
Adeline (HPLC) and Mrs. Gaspard (Service des Cultures
Cellulaires, ICSN).
by CH
Na SO
gave compound 12 (7.2 mg, yield 67%) as an amorphous
2
Cl
2
. The combined organic phases were washed, dried
(
2
4
), and evaporated. The residue, after preparative TLC,
2
5
powder: [R]
D
-118° (c 0.12, CHCl ); UV (MeOH) λmax (log ꢀ)
3