New Ellagitannins from Phyllanthus
J ournal of Natural Products, 2001, Vol. 64, No. 12 1531
cm- ; H NMR (acetone-d
H-G2,6), 7.13 (1H, s, H-D ), 7.03 (1H, s, HHDP H-A
s, HHDP H-B ), 6.44 (1H, br s, H-1), 6.03 (1H, br s, H-2), 5.40
1H, br s, H-4), 5.21 (1H, d, J ) 3.5 Hz, H-3), 4.83 (1H, d, J )
.5 Hz, H-E ), 4.82 (1H, m, H-5), 4.81 (1H, dd, J ) 2.5, 12.0
Hz, H-6a), 4.40(1H, dd, J ) 7.0, 12.0 Hz, H-6b), 3.73 (1H, td,
1
1
, 500 MHz) δ 7.16 (2H, s, galloyl
), 6.68 (1H,
P h ylla n em blin in D (4): white amorphous powder; [R]22
6
D
3
3
-7.9° (c 0.33, MeOH); UV (MeOH) λmax (log ꢀ) 282 (4.16) nm;
3
IR (DR) νmax 3382, 1717, 1614, 1528, 1452, 1376, 1313, 1194,
-
(
2
1067 cm 1; 1H NMR (acetone-d
6
, 500 MHz) δ 7.21 (2H, s,
3
galloyl H-2′, 6′), 7.15 (1H, s, neochebuloyl H-3′′), 5.82 (1H, d,
J ) 8.0 Hz, H-1), 5.42 (1H, d, J ) 1.0 Hz, neochebuloyl H-2′′′),
5.28 (1H, t, J ) 9.5 Hz, H-3), 3.98 (1H, dd, J ) 1.0, 10.5 Hz,
H-3′′′), 3.91 (1H, dd, J ) 2.0, 12.0 Hz, H-6a), 3.83 (1H, dd, J
) 8.0, 9.5 Hz, H-2), 3.79 (1H, dd, J ) 4.5, 12.0 Hz, H-6b), 3.77
(1H, dd, J ) 6.0, 9.5 Hz, H-4), 3.69 (1H, ddd, J ) 2.0, 4.5, 6.0
Hz, H-5), 3.22 (1H, ddd, J ) 4.5, 10.5, 11.0 Hz, H-4′′′), 3.01
(1H, dd, J ) 11.0, 17.5 Hz, H-5′′′a), 2.49 (1H, dd, J ) 4.5, 17.5
J ) 2.5, 12.0 Hz, H-E
4
), 2.65 (1H, dd, J ) 2.5, 17.5 Hz, H-E5a),
1
3
1
7
(
C-G
(
6
.96 (1H, dd, J ) 12.0, 17.5 Hz, H-E5b); C NMR (acetone-d ,
5 MHz) δ 173.7 (C-E
HHDP C-A
6
), 173.4 (C-E
7
), 171.0 (C-E
), 165.2 (galloyl
), 145.1, 145.3
), 137.6 (C-B ),
), 120.6 (C-
1
), 168.6
7
), 165.0 (C-D
7
), 166.4 (HHDP C-B
7
7
), 147.2 (C-D
A
4
), 146.6 (C-D
6
), 145.9 (C-G3, G
5
C-A4,
6
), 144.8 144.5 (C-B4,
36.3 (C-A ), 135.7 (C-D ), 125.3 (C-A
), 119.9 (C-G ), 117.2 (C-D ), 117.0 (C-B
C-D ), 110.5 (C-G2, ), 110.2 (C-B
), 92.0 (C-1), 73.4 (C-5), 69.7 (C-2), 66.3 (C-4), 63.9 (C-6),
1.3 (C-3), 49.2 (C-E ), 41.5 (C-E ), 30.9 (C-E ); FABMS m/z
69 [M - H] ; anal. C 46.80%, H 4.06%, calcd for C41
/2H O, C 46.82%, H 3.74%.
Meth yla tion of 3. A mixture of compound 3 (50 mg),
dimethyl sulfate (1 mL), and anhydrous potassium carbonate
1 g) in dry acetone (10 mL) was refluxed for 1 h with stirring.
B
6
), 139.8 (C-G
4
5
1
5
5
2
), 124.4 (C-B
2
Hz, H-5′′′b); 13C NMR (acetone-d
6
, 75 MHz): δ 174.7, 174.3,
D
2
1
1
1
), 115.3 (C-A
1
), 112.1
171.2, 165.6, 164.4 (neochebuloyl C-1′′′, 6′′′, 7′′′, 7′′, galloyl
C-7′), 146.1 (C-3′, 5′, 6′′), 143.5 (C-4′′), 139.5 (C-5′′), 139.2 (C-
4′), 120.6 (C-1′), 117.3 (C-2′′), 116.7 (C-1′′), 110.3 (C-2′, 6′), 109.4
(C-3′′), 95.1 (C-1), 79.4 (C-2′′′), 78.2 (C-5), 77.7 (C-3), 72.1 (C-
2), 68.8 (C-4), 61.9 (C-6), 46.0 (C-3′′′), 36.8 (C-4′′′), 35.1 (C-
(
3
G
6
3
), 107.8 (C-E2), 107.5 (C-
A
3
6
9
9
3
4
5
-
30 28
H O ‚
-
2
5′′′); FABMS m/z 669 [M - H] (33), 153 (100); anal. C 44.95%,
H 4.45%, calcd for C27
H
26
O
2
20‚3H O, C 44.76%, H 4.45%.
Hyd r olysis of 4 w ith Ta n n a se. A solution of 4 (10 mg) in
O (1 mL) was incubated with tannase (1 mg) at 37 °C for 2
h. The reaction mixture was separated by chromatography
over Sephadex LH-20 with H O-MeOH to give the hydrolysate
(4a ) (5.4 mg) and gallic acid (2.0 mg). 4a : pale amorphous
(
H
2
After removal of the inorganic compounds by filtration, the
filtrate was concentrated to dryness. The residue was applied
to a column of silica gel eluting with toluene-acetone (5:1) to
give the methyl derivative (3a ) (39 mg): white amorphous
powder, [R]
754, 1590, 1464, 1343, 1223, 1180, 1027 cm-1
CDCl , 500 MHz) δ 7.21 (2H, s, H-G2, ), 7.19 (1H, s, H-D
.74 (1H, s, H-A ), 6.67 (1H, s, H-B ), 6.51 (1H, br s, H-1), 5.95
2
powder, [R]24
1
D
6
+29.9° (c 0.57, MeOH); H NMR (acetone-d ,
2
4
D
-57.2° (c 0.31, CHCl
3
); IR (DR) νmax 2944, 2844,
300 MHz) δ 7.14 (1H, br s, R and â form, neochebuloyl H-3′′),
5.40 (1H, br s, R and â form, neochebuloyl H-2′′′), 5.38 (1/2H,
dd, J ) 7.8, 9.9 Hz, R form-glu H-3), 5.25 (1/2H, d, J ) 3.9 Hz,
R form-glu H-1), 5.12 (1/2H, dd, J ) 9.3, 9.6 Hz, â form-glu
H-3), 4.70 (1/2H, d, J ) 7.8 Hz, â form-glu H-1), 3.22 (1H, m,
R and â form H-4′′′), 2.97 (1H, dd, J ) 10.5, 16.5 Hz, R and â
form H-5′′′a), 2.44 (1H, br d, J ) 16.5 Hz, R and â form H-5′′′b).
1
1
(
;
H NMR
),
3
6
3
6
3
3
(
1H, br s, H-3), 5.41 (1H, br s, H-2), 5.38 (1H, br t, J ) 2.0 Hz,
H-4), 5.04 (1H, t, J ) 11.0 Hz, H-6a), 4.85 (1H, ddd, J ) 2.0,
.0, 11.0 Hz, H-5), 4.78 (1H, d, J ) 2.5 Hz, H-E ), 4.45 (1H,
dd, J ) 8.0, 11.0 Hz, H-6b), 4.12 (3H, s, OMe-D ), 3.97, 3.96,
.96, 3.95 (each 3H, s, OMe-A , A , B , G ), 3.90 (3H, s, OMe-
), 3.87 (3H, s, OMe-E ), 3.86 (1H, td, J ) 2.5, 11.5 Hz, H-E
.71, 3.69 (each 3H, s, OMe-A , B ), 3.68 (6H, s, OMe-G3,
.63 (3H, s, OMe-E ), 3.61 (3H, s, OMe-E
), 2.46 (1H, dd, J ) 2.5, 17.0 Hz, H-E5a), 2.07 (1H, dd, J )
8
3
2
2
5
P h ylla n em blin in E (5): white amorphous powder; [R]
D
3
4
5
4
4
-8.3° (c 0.36, MeOH); UV (MeOH) λmax (log ꢀ) 282 (4.22) nm;
D
3
3
4
1
4
5
),
),
IR (DR) νmax 3346, 1733, 1618, 1452, 1388, 1348, 1228, 1120,
1081 cm 1; 1H NMR (acetone-d
-
, 500 MHz) δ 7.20 (2H, s,
6
6
6
2
6
), 3.23 (3H, s, OMe-
galloyl H-2′, 6′), 7.13 (1H, s, neochebuloyl H-3′′), 5.78 (1H, d,
J ) 8.5 Hz, H-1), 5.66 (1H, d, J ) 1.0 Hz, neochebuloyl H-2′′′),
5.05 (1H, dd, J ) 9.5, 10.0 Hz, H-4), 3.90 (1H, dd, J ) 9.0, 9.5
Hz, H-3), 3.88 (1H, dd, J ) 1.0, 10.5 Hz, H-3′′′), 3.79 (1H, J )
2.0, 6.0, 9.5 Hz, H-5), 3.77 (1H, dd, J ) 2.0, 12.5 Hz, H-6a),
3.67 (1H, dd, J ) 8.5, 9.0 Hz, H-2), 3.63 (1H, dd, J ) 6.0, 12.5
Hz, H-6b), 3.14 (1H, ddd, J ) 3.0, 10.5, 12.0 Hz, H-4′′′), 3.07
(1H, dd, J ) 12.0, 17.0 Hz, H-5′′′a), 2.37 (1H, dd, J ) 3.0, 17.0
B
1
4
1
3
1.5, 17.0 Hz, H-E5b); C NMR (CDCl
3
, 75 MHz) δ 172.5 (C-
), 167.6 (C-A ), 164.6 (C-B ), 164.2
), 153.3 (C-G3, 5), 153.2 (C-A ), 152.7, 152.6,
, A , B , B ), 148.3 (C-D ), 145.3 (C-B ), 145.0
), 143.2 (C-G ), 138.1 (C-D ), 128.3 (C-A ), 126.0 (C-B ),
), 123.1 (C-G ), 121.3 (C-A ), 120.1 (C-D ), 119.5
), 108.8 (C-D ), 107.6 (C-B ), 106.9 (C-G2,6),
), 92.6 (C-1), 72.4 (C-5), 67.7 (C-2), 65.0 (C-4), 63.4
, A , B , A , B
), 55.3 (OMe-B ), 54.9
), 49.4 (C-E ), 40.0 (C-
); FABMS m/z 1165 [M - H] (10); anal. C
6.94%, H 5.23%, calcd for C55 28, C 56.61%, H 5.01%.
Alk a lin e Tr ea tm en t of 3a , F ollow ed by Meth yla tion .
A solution of 3a (39 mg) in 5% NaOH [(H
E
6
), 171.8 (C-E
C-G
7
), 168.7 (C-E
1
7
7
(
7
), 163.6 (C-D
7
4
1
52.5, 152.3 (C-D
4
6
4
6
6
5
(
C-A
5
4
5
2
2
1
24.2 (C-B
C-D
04.8 (C-A
1
1
1
1
(
2
), 110.6 (C-E
2
3
3
Hz, H-5′′′b); 13C NMR (acetone-d
6
, 75 MHz) δ 174.4, 174.2,
1
3
171.3, 165.9, 164.6 (neochebuloyl C-1′′′, 6′′′, 7′′′, 7′′, galloyl
C-7′), 146.1 (C-6′′), 146.0 (C-3′, 5′), 143.4 (C-4′′), 139.5 (C-5′′),
139.2 (C-4′), 120.5 (C-1′), 117.3 (C-2′′), 116.4 (C-1′′), 110.2 (C-
2′, 6′), 109.3 (C-3′′), 95.3 (C-1), 77.4 (C-2′′′), 76.4 (C-5), 75.1
(C-3), 74.1 (C-2), 72.0 (C-4), 61.6 (C-6), 45.8 (C-3′′′), 37.0 (C-
(
C-6), 61.1, 60.9, 60.8, 60.7, 60.6 (C-3, OMe-D
), 56.5 (OMe-G3,5, D ), 56.0 (OMe-A
OMe-E ), 53.4 (OMe-E ), 51.7 (OMe-E
), 29.9 (C-E
5
6
6
5
5
,
G
4
4
4
4
(
E
2
1
6
3
-
4
5
-
5
H
58
O
4′′′), 34.9 (C-5′′′); FABMS m/z 669 [M - H] (20), 517 (7), 153
(100); anal. C 44.56%, H 4.43%, calcd for C27
44.76%, H 4.45%.
H
26
O
2
20‚3H O, C
2
O-MeOH (1:1)) (2
mL) was heated at 70 °C for 30 min. The reaction mixture
was acidified with 1 M HCl (10 mL) and extracted with ether
three times. The organic layer was dried over Na SO , con-
2 4
centrated to dryness, and treated with ethereal diazomethane.
After evaporation of the solvent, the syrupy residue was
chromatographed over silica gel with toluene-acetone (5:1),
furnishing methyl trimethoxybenzoate (3b) (7.4 mg), (R)-
Hyd r olysis of 5 w ith Ta n n a se. A solution of 5 (10 mg) in
O (1 mL) was incubated with tannase (1 mg) at 37 °C for 2
h. The reaction mixture was separated by chromatography
over Sephadex LH-20 with H O-MeOH to afford the hydroly-
sate (5a ) (5.0 mg) and gallic acid (3.0 mg). 5a : pale amorphous
H
2
2
powder, [R]24
1
D
6
+7.7° (c 0.28, MeOH); H NMR (acetone-d , 300
MHz) δ 7.13, 7.12 (each 1/2H, s, R and â form neochebuloyl
H-3′′), 5.59 (1H, br s, R and â form, neochebuloyl H-2′′′), 5.21
(1/2H, br s, R form glu H-1), 4.88 (1H, br dd, J ) 7.5, 11.1 Hz,
R and â form H-3), 4.63 (1/2H, d, J ) 8.4 Hz, â form H-1), 4.13
(1H, m, R and â form H-3′′′), 3.06 (1H, m, R and â form H-4′′′),
2.99 (1H, m, R and â form H-5′′′a), 2.32 (1H, br d, J ) 15.6
2
2
dimethyl hexamethoxydiphenoate (3c) (9.1 mg), [R]
D
+15.2°
2
8
(
c 0.46, CHCl
3
), and 3d (5.1 mg), [R]
H NMR (CDCl , 500 MHz) δ 7.11 (1H, s, H-D
J ) 2.5 Hz, H-E ), 4.08 (3H, s, OMe-D ), 3.89 (3H, s, OMe-D
.83 (3H, s, OMe-D ), 3.80 (3H, s, OMe-E
2.5, 3.5, 11.0 Hz, H-E ), 3.64 (3H, s, OMe-E
), 3.55 (3H, s, OMe-E ), 2.57 (1H, dd, J ) 11.0, 17.0
D
-65.9° (c 0.12, CHCl
3
);
1
3
3
), 4.41 (1H, d,
3
5
4
),
3
)
7
1
), 3.75 (1H, ddd, J
), 3.62 (3H, s,
Hz, R and â form H-5′′′b). The [R]
were identical to those of 4-O-neochebuloyl-D-glucose, which
and 1H NMR data of 5a
D
4
6
OMe-E
7
2
1
3
Hz, H-E5a), 2.41 (1H, dd, J ) 3.5, 17.0 Hz, H-E5b); C NMR
CDCl , 125 MHz) δ 173.4 (C-E ), 172.7 (C-E ), 170.0 (C-E ),
66.0 (C-D ), 152.1 (C-D ), 148.9 (C-D ), 137.0 (C-D ), 122.8
C-D ), 120.7 (C-D ), 110.5 (C-E ), 107.2 (C-D ), 60.6 (OMe-
), 56.4 (OMe-D ), 54.3 (OMe-E ), 53.1 (OMe-E ), 52.1 (OMe-
), 51.9 (OMe-E ), 51.7 (OMe-E ), 50.3 (C-E ), 39.8 (C-E ), 33.3
); EIMS m/z 470 [M] .
was prepared from chebulinic acid by refluxing in H
followed with tannase hydrolysis.
2
O for 2 h
(
3
7
6
1
2
2
1
7
4
6
5
P h ylla n em blin in F (6): white amorphous powder; [R]
D
(
1
2
2
3
-18.3° (c 0.17, MeOH); UV (MeOH) λmax (log ꢀ) 281 (4.14) nm;
IR (DR) νmax 3343, 1715, 1617, 1528, 1452, 1342, 1204, 1063
D
D
(
5
7
4
2
1
-
1 1
7
6
3
4
cm ; H NMR (acetone-d
6
, 300 MHz) δ 7.18(2H, s, galloyl H-2′,
+
C-E
5
6′), 7.14(1H, s, neochebuloyl H-3′′), 5.70 (1H, d, J ) 7.8 Hz,