1033
Journal of Natural Produm Vol. 55. N o . 8 . p p . 1033-1043. August 1992
ANTITUMOR AGENTS, 129. TANNINS AND RELATED COMPOUNDS AS SELECTIVE CYTOTOXIC AGENTS YOSHIKIKASHIWADA,
Natural Products Laboratory, Division of Medicinal Chemistry and Natural Products, Schwl of Pharmasy, University of North Carolina, Chapel Hill, North Carolina 27599 GEN-ICHIRO NONAKA,ITSUO NISHIOKA,
Faculty of Pharmaceutical Sciences, Kyushu Uniwrsity, Fukuoh 812, Japan JER-JANGCHANG,
Department of Pathology, Srhwl of Medicine, University of North Carolina, Chapel Hill, North Carolina 27599 and KUO-HSIUNGLEE*
Natural Products Laboratwy, Division of Medicinal Chemistry and Natural P d c t s , Schwl of Pharmasy, University of Nwth Carolina, Chapel Hill, Nwth Carolina 27599 ABSTRACT.-Fifty-seven tannins and related compounds, including gallotannins, ellagitannins, and condensed and complex tannins, were evaluated for their cytotoxicities against human tumor cell lines, including malignant melanoma, lung carcinoma, ileocecal adenocarcinoma, epidermoid carcinoma, malignant melanoma, and medulloblastoma cell lines. Among them, chebulagic acid 111, geraniin 123, sanguiin H-1 1 131,4,5di-O-galloylquinic acid 1121, 1,3,4,5-tetra-0-galloylquinicacid [15], 1(P)-O-galloylpedunculagin 1241, furosin 1291, castalagin [%], sanguiin H-2 1341, vexalagin 1391, grandinin 1401, phyllyraeoidin A 1421, (-)epicatechin 3-0-gallate [SO], cinnamtannin B, 1551, and acutissimin A [%I exhibited mcderate selective cytotoxicity against PRMI-795 1 melanoma cells with ED,, values in the range of 0.1-0.8 p,g/ml. Selective cytotoxicities against the melanoma cells were also observed for strictinin 1221, pedunculagin [23], eugeniin 1251, elaeocarpusin 1281, punicacortein C 137, casuarinin 1411, sanguiin H-6 1431, procyanidin B-2 3,3’-di-O-gallate 1511, procyanidin C- 1 3,3’,3”-tri-O-gallate 1521, and cinnamtannin B, 1541 with ED,, values of 1 4 p,g/ml. All of the tannins were found to be inactive (>10 pglml) against lung carcinoma (A-549),ileocecal adenocarcinoma (HCT-8), epidermoid carcinoma of nasopharnyx (KB), and medulloblastoma (TE-67 1) tumor cells.
Tannins are widely distributed in the plant kingdom and are classified on the basis of their structures into three groups: hydrolyzable, condensed, and complex tannins. HO H
HO OH
O
W
o=c
1 ‘For part 128, see Lee et ai. (1).
OH
O
c=o
2
H
Wol. 5 5 , No. 8
Journal of Natural Products
1034
3
They are known to possess a variety of biological activities, such as lowering the level of blood urea nitrogen (2,3), inhibiting the activity of angiotensin-converting enzyme ( 4 ) , antiviral and anti-HIV activities (5,6), inhibition of lipid peroxidation (7,8), antimutagenic effects (9), DNA-breaking activity ( 1 0 , l l), and psychotropic activity (12). h v e s of Tewninalia chbula (100 g ) 70% aqueous Me,CO concentrated extract
+10
+>10
+>10
+>10
f C 10
MCI-gel CHPZOP Chromatography [HZO-MeOH ( 1 :OH 1 l)]
I +10
Fuji-gel ODS-Q3 [HZO-MeOH (1:OHl:l)] chebulagic acid (530 mg) +0.80
SCHEME 1
*: ED,, value in kg/ml against melanoma RPMI-795 1
August 1992)
Kashiwada et ai.: Tannins as Cytotoxic Agents
1035
As a result of our continuing search among medicinal plants for novel potent selective cytotoxic agents against solid tumors, the alcoholic extracts of the leaves of Terminalia chebula (Combretaceae), the whole plants of Geranium thunbergii (Geraniaceae), and the roots of Sanguiswba ofiinalis (Rosaceae)were found to show significant (ED,, < 20 kg/ml) cytotoxicity in melanoma tumor cells (RPMI-795 1). Subsequent bioassaydirected fractionation led to the isolation of known tannins, chebulagic acid 111,geraniin 121,and sanguiin H- 11 131 from T. cbebula, G. thunbwgii, and S . ofiinalis, respectively, as the cytotoxic principles (see Schemes 1-3). Since 1-3 are the first identified tannins to demonstrate potent cytotoxicity against this human cancer cell line, it is important to screen the other tannins as potent selective cytotoxic agents against solid tumor cell lines, although the antitumor activity of tannins against sarcoma-180 has previously been reported (13).We report herein cytotoxicity determinations of fiftyseven tannins against lung carcinoma (A-549), ileocecal adenocarcinoma (HCT-8), epidermoid carcinoma of nasopharnyx (KB), melanoma (RPMI-795 l),and medulloblostoma (TE-67 1) tumor cells. Roots of Sanguisorba offiriinalis (50 g) 70% aqueous Me,CO concentrated extract f10
Sephadex LH-20 chromatography
50% aqueous Me,CO
+>10
f >10
+>10
+10
+>10
+>10
' 10 > 10 > 10 > 10 > 10 > 10 5.01
> 10 0.53
> 10 > 10 0.43
> 10 > 10 > 10 > 10 > 10
Ellagitannins
1 2 3 21 22 23 24 25 26 27 28 29 30 31 32 33
. . . .
. . . .
. . . .
. . . .
. . . .
. . . .
. . . .
.......
. . . . . . .
. . . . . . . . . . . . . . . . . . . . .
. . . . . . . 34 . 35 .
. . . . . . . . .
. . . . . . . . .
. . . . . . . . .
. . . . . . . . .
. . . . . . . . . . . . . . . . . .
36 . . . . . . . 37 . . . . . . . 38 . . . . . . . 39 . . . . . . .
40 . . . . . . . . . . . . .
41 . 42 . 43 . %. 45 .
. . . . . .
. . . . . . . . . . . . . . . . . .
chebulagicacid (35) geraniin(31) sanguiin H- 11 (26) 2.3.hexahydroxydiphenoyl. ~.glucose(26) strictinin( 17) pedunculagin (27) l(P)-O-galloylpedunculagin(28) eugeniin(29) corilagin (30) punicafolin (30) elmarpusin (32) furosin (33) terchebin (33) chebulinic acid (34) mallotinicacid (33) mallotusinic acid (33) sanguiin H-2 (26) punicalin (36) punicalagin (36) punicacorteinC(37) castalagin (38) vescalagin (38) grandinin (38. 39) casuarinin (38) phillyraeoidin A (40) sanguiin H-6 (26) castanopsininA (40) castanopsiniinA (40)
CondensedTannins
46 . 47 . 48 . 49 . 50 . 51 . 52 . 53 .
. . . . . . .
. . . . . . .
. . . . . . .
. . . . . . .
. . . . . . .
. . . . . . .
(+)-catechin (42) (-)-epicarechin (43) procyanidin B-2 (43) procyanidin C- 1 (43) (-)-epicatechin 3-0-gallate(42) procyanidinB.23.3I.di. O.gallate(42) procyanidinC.l3.3’.3”.tri. O.gallate(42) . . . . . . proanthocyanidin A-2 (44)
0.80 0.35 0.50 > 10 4.86 2.74 0.21 2.74 > 10 > 10 2.62 0.67 > 10 > 10 > 10 > 10 0.44 > 10 > 10 3.86 0.79 0.58 0.10 2.24 0.50 5 .00 > 10 > 10
> 10 > 10 > 10 > 10 0.55 3.45 3.05 5.50
1042
Journal of Natural Products
Compound
54 . . . . 55 . . . .
.. . .. .
Name cinnamtannin B, (43) cinnamtannin B, (43)
[Vol. 55, No. 8
ED,, (kg/ml) 3.57 0.66
Complex Tannins
56 . . . . . . . acutissimin A (38,45) 57
.. .....
mongolicainA(38,46) Actinomycin (positive control)
0.61
> 10 c0.01
MATERIALSAND METHODS MATERIhLS.--The 57 tannins and related compounds were isolated from the plant materials as reported previously (14-46) and shown in Schemes 1-3. BIOLOGICAL ASAY.--The in vitro cytotoxicity assay was carried out according to a National Cancer Institute protocol (47) as previously described (48). The assay was conducted using apanel ofhuman tumor cell lines. These cell lines are lung carcinoma (A-549), ileocecal adenocarcinoma(HCT-8), epidermoid carcinomaof nasopharnyx (KB), melanoma (RPMI-795 I), and medulloblastoma(TE-67 1). All cell lines were obtained from the American Type Culture Collection, Rockville, MD. In general, the assay methods were the same as those described by Monks et ul. (49). However, a tetrazolium salt method was used to measure cell survival and proliferation (50). ACKNOWLEDGMENTS This work was supported by grant from the National Cancer Institute No. CA-17625 (K.H. Lee).
LITERATURE CITED 1. K.H. Lee, J.X. Xie, and H. Hu, Abstracts ofchinere Medicine, 4 , 4 7 3 (1991). 2. T. Nagasawa, H. Oura, Y. Shoyama, and I. Nishioka, Chem. P h n n . Bull., 28, 1736 (1980). 3. S. Shibutani, T. Nagasawa, H. Oura, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 31,2378 (1983). 4. J. Inokuchi, H. Okabe, T. Yamauchi, A. Nagamatsu, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 33,264 (1985). 5. M.Nishizawa, T. Yamagishi, G.E. Dutschman, W.B. Parker, A.J. Bodner, R.E.Kilkuskie, Y.C. Cheng, and K.H. Lee,]. Nut. Prod., 52,762 (1989). 6. G. Nonaka, I. Nishioka, M. Nishizawa, T. Yamagishi, Y. Kashiwada, G.E. Dutschman, A.J. Bodner, R.E. Kilkuskie, Y.C. Cheng, and K.H. Lee,j.Nut. Prod., 53,587 (1990). 7. Y. Kimura, H. Okuda, T. Okuda, T. Hatano, I. Agata, and S.Archi, Plantu'Med.,50,473 (1984). 8. Y. Kimura, H. Okuda, T. Okuda, T. Hatano, I. Agata, and S. Archi, Chem. Phunn. BuII., 33, 2028 (1985). 9. T.Gichner, F. Posiisil,J. Veleminsky, V. Volkeova, and J. Volke, FoliuMirrobiol. (Prugue), 32,55 (1987). 10. S. Shirahata, H. Murakami, K. Nishiyama, I. Sugata, K. Shinohara, G. Nonaka, I. Nishioka, and H. Omura, Agrir. Biol. Chem., 49, 1033 (1985). 11. S. Shirahata, H . Murakami, K. Nishiyama, K. Yamada, G. Nonaka, I. Nishioka, and H. Omura, J . Agric. F w d C h . , 37,299 (1989). 12. S. Ueki, G. Nonaka, I. Nishioka, and M. Fujiwara, J . Med. Phunn. Sor. Wukan-YuLu, 2 , 502 (1985). 13. K. Miyamoro, N. Kishi, R. Koshiura, T. Yoshida, T. Hatano, and T.Okuda, Chem. Phunn. Bull., 35,814 (1987). 14. Y. Kashiwada, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 32,3461 (1984). 15. G. Nonakaand I. Nishioka, Chem. Phann. Bull., 31, 1652 (1983). 16. G. Nonaka, I. Nishioka, T.Nagasawa, and H. Oura, C h . Phunn. Bull., 29,2862 (1981). 17. G. Nonaka, R. Sakai, and I. Nishioka, Phytorhemistry, 23, 1753 (1984). 18. M. Nishizawa, T. Yamagishi, G. Nonaka, and I. Nishioka, J. Chem. Sor., Perkin Trans. 1 , 961 (1983).
August 19927 19. 20. 2 1. 22. 23.
24. 25. 26. 27. 28. 29. 30. 31. 32. 33. 34. 35. 36. 37. 38. 39. 40. 41. 42. 43. 44. 45. 46. 47. 48. 49.
50.
Kashiwada et al.: Tannins as Cytotoxic Agents
1043
G. Nonaka, I. Ishirnatsu, T. Tanaka, I. Nishioka, M. Nishizawa, and T. Yrnamagishi, Chem. Phunn. Bull., 35, 3127 (1987). M. Nishizawa, T. Yamagishi, G. Nonaka, and I. Nishioka,]. Chn. Soc., Perkin Trans. 1 , 2968 (1982). H. Nishimura, G. Nonaka, and I. Nishioka, Phytwhzstry, 23, 262 1 (1984). K. Ishirnaru, G. Nonaka, and I. Nishioka, P b y t o r h i m y , 25, 1501 (1987). M. Nishizawa, T. Yamagishi, G. Dutxhman, W.B. Parker, A.J. Bodner, R.E. Kilkuskie, Y.C. Cheng, and K.H. Lee,]. Nut. Prod., 5 2 , 762 (1989). G. Nonaka, M. Ageta, and I. Nishioka, Chnn. Phann. Bull., 36, 96 (1985). G. Nonaka, K. Ishirnaru, T. Tanaka, and I. Nishioka, Chem. Phum. Bull., 3 2 , 4 8 3 (1984). T. Tanaka, G. Nonaka, and I. Nishioka,]. C h . Res., Synop., 176 (1985);]. C h . Rer., Miniprint, 2001 (1985). H. Feng, G. Nonaka, and I. Nishioka, Pbytorhnnistry, 27, 1185 (1988). G. Nonaka, M. Ishimatsu, M. Ageta, and I. Nishioka, Chem. Phunn. Bull., 37, 50 (1989). G. Nonaka, M. Harada, and I. Nishioka, Chem. Phunn. Bull., 28, 685 (1980). T. Tanaka, G. Nonaka, and I. Nishioka, Phytwhemistry, 24, 2075 (1985). T. Tanaka, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 34, 941 (1986). T. Tanaka, G. Nonaka, I. Nishioka, K. Miyahara, and T. Kawasaki,]. Chem. Soc., Perkin Trans. I , 369 (1986). R. Saijo, G. Nonaka, and I. Nishioka, Chem. Pburm. Bull., 37, 2063 (1989). T.C. Lin, G. Nonaka, I. Nishioka, and F.C. Ho, Chem. Phunn. Bull., 38, 3004 (1990). T. Tanaka, G. Nonaka, andI. Nishioka, Chem. Pbunn. Bull., 34, 1039(1986). T. Tanaka, G. Nonaka, and I. Nishioka, Chem. Phann. Bull., 34,650 (1986). T. Tanaka, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 34,656 (1986). G. Nonaka, T. Sakai, T. Tanaka, K. Mihashi, and I. Nishioka, Chem. Pbunn. Bull., 38, 2151 (1990). G. Nonaka, K. Ishirnaru, R. Azuma, M. Ishirnatsu, and I. Nishioka, Chem. Phunn. Bull., 37,207 1 (1989). G. Nonaka, S. Nakayama, and I. Nishioka, Chem.Phunn. Bull., 37, 2030 (1989). M. Ageta, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 36, 1646 (1988). Y. Kashiwada, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 34,4083 (1986). S. Morimoto, G. Nonaka, and I. Nishioka, Chem. Pbunn. Bull., 34, 633 (1986). G. Nonaka, S. Morirnoto, and I. Nishioka,]. Chem. Soc., Perkin Truns. 1 , 2 139 (1983). K. Ishirnaru, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 35, 602 (1987). G. Nonaka, K. Ishirnaru, K. Mihashi, Y. Iwase, M. Ageta, and I. Nishioka, Chem. Pbunn. Bull., 36, 857 (1988). R.I. Geran, N.H. Greenberg, N.M. McDonald, A.M. Schumacher, and B.J. Abbott, Cancer Chemotber. Rep., Part 3, 3, 1 (1972). K.H. Lee, Y.M. Lin, T.S. Wu, D.C. Zhang, T. Yamagishi, T. Hayashi, I.H. Hall, J.J. Chang, R.Y. Wu, and T.H. Yang, PIunta Med., 54, 308 (1988). A. Monks, D. Scudiero, P. Skehan, R. Shoemaker, K. Paull, D. Vistica, C. Hose, J. Langley, P. Cronise, A. Vaigro-Wolff, M. Gray-Goodrich, H. Campbell, J . Mayo, and M. Boyd,]. Nutl. Cunrerlnst., 83, 757 (1991). T. Mosrnann,J . fmmunol. Methodr, 65, 55 (1983).
Received 17 Ortober 1991
Journal of Natural Produm Vol. 55. N o . 8 . p p . 1033-1043. August 1992
ANTITUMOR AGENTS, 129. TANNINS AND RELATED COMPOUNDS AS SELECTIVE CYTOTOXIC AGENTS YOSHIKIKASHIWADA,
Natural Products Laboratory, Division of Medicinal Chemistry and Natural Products, Schwl of Pharmasy, University of North Carolina, Chapel Hill, North Carolina 27599 GEN-ICHIRO NONAKA,ITSUO NISHIOKA,
Faculty of Pharmaceutical Sciences, Kyushu Uniwrsity, Fukuoh 812, Japan JER-JANGCHANG,
Department of Pathology, Srhwl of Medicine, University of North Carolina, Chapel Hill, North Carolina 27599 and KUO-HSIUNGLEE*
Natural Products Laboratwy, Division of Medicinal Chemistry and Natural P d c t s , Schwl of Pharmasy, University of Nwth Carolina, Chapel Hill, Nwth Carolina 27599 ABSTRACT.-Fifty-seven tannins and related compounds, including gallotannins, ellagitannins, and condensed and complex tannins, were evaluated for their cytotoxicities against human tumor cell lines, including malignant melanoma, lung carcinoma, ileocecal adenocarcinoma, epidermoid carcinoma, malignant melanoma, and medulloblastoma cell lines. Among them, chebulagic acid 111, geraniin 123, sanguiin H-1 1 131,4,5di-O-galloylquinic acid 1121, 1,3,4,5-tetra-0-galloylquinicacid [15], 1(P)-O-galloylpedunculagin 1241, furosin 1291, castalagin [%], sanguiin H-2 1341, vexalagin 1391, grandinin 1401, phyllyraeoidin A 1421, (-)epicatechin 3-0-gallate [SO], cinnamtannin B, 1551, and acutissimin A [%I exhibited mcderate selective cytotoxicity against PRMI-795 1 melanoma cells with ED,, values in the range of 0.1-0.8 p,g/ml. Selective cytotoxicities against the melanoma cells were also observed for strictinin 1221, pedunculagin [23], eugeniin 1251, elaeocarpusin 1281, punicacortein C 137, casuarinin 1411, sanguiin H-6 1431, procyanidin B-2 3,3’-di-O-gallate 1511, procyanidin C- 1 3,3’,3”-tri-O-gallate 1521, and cinnamtannin B, 1541 with ED,, values of 1 4 p,g/ml. All of the tannins were found to be inactive (>10 pglml) against lung carcinoma (A-549),ileocecal adenocarcinoma (HCT-8), epidermoid carcinoma of nasopharnyx (KB), and medulloblastoma (TE-67 1) tumor cells.
Tannins are widely distributed in the plant kingdom and are classified on the basis of their structures into three groups: hydrolyzable, condensed, and complex tannins. HO H
HO OH
O
W
o=c
1 ‘For part 128, see Lee et ai. (1).
OH
O
c=o
2
H
Wol. 5 5 , No. 8
Journal of Natural Products
1034
3
They are known to possess a variety of biological activities, such as lowering the level of blood urea nitrogen (2,3), inhibiting the activity of angiotensin-converting enzyme ( 4 ) , antiviral and anti-HIV activities (5,6), inhibition of lipid peroxidation (7,8), antimutagenic effects (9), DNA-breaking activity ( 1 0 , l l), and psychotropic activity (12). h v e s of Tewninalia chbula (100 g ) 70% aqueous Me,CO concentrated extract
+10
+>10
+>10
+>10
f C 10
MCI-gel CHPZOP Chromatography [HZO-MeOH ( 1 :OH 1 l)]
I +10
Fuji-gel ODS-Q3 [HZO-MeOH (1:OHl:l)] chebulagic acid (530 mg) +0.80
SCHEME 1
*: ED,, value in kg/ml against melanoma RPMI-795 1
August 1992)
Kashiwada et ai.: Tannins as Cytotoxic Agents
1035
As a result of our continuing search among medicinal plants for novel potent selective cytotoxic agents against solid tumors, the alcoholic extracts of the leaves of Terminalia chebula (Combretaceae), the whole plants of Geranium thunbergii (Geraniaceae), and the roots of Sanguiswba ofiinalis (Rosaceae)were found to show significant (ED,, < 20 kg/ml) cytotoxicity in melanoma tumor cells (RPMI-795 1). Subsequent bioassaydirected fractionation led to the isolation of known tannins, chebulagic acid 111,geraniin 121,and sanguiin H- 11 131 from T. cbebula, G. thunbwgii, and S . ofiinalis, respectively, as the cytotoxic principles (see Schemes 1-3). Since 1-3 are the first identified tannins to demonstrate potent cytotoxicity against this human cancer cell line, it is important to screen the other tannins as potent selective cytotoxic agents against solid tumor cell lines, although the antitumor activity of tannins against sarcoma-180 has previously been reported (13).We report herein cytotoxicity determinations of fiftyseven tannins against lung carcinoma (A-549), ileocecal adenocarcinoma (HCT-8), epidermoid carcinoma of nasopharnyx (KB), melanoma (RPMI-795 l),and medulloblostoma (TE-67 1) tumor cells. Roots of Sanguisorba offiriinalis (50 g) 70% aqueous Me,CO concentrated extract f10
Sephadex LH-20 chromatography
50% aqueous Me,CO
+>10
f >10
+>10
+10
+>10
+>10
' 10 > 10 > 10 > 10 > 10 > 10 5.01
> 10 0.53
> 10 > 10 0.43
> 10 > 10 > 10 > 10 > 10
Ellagitannins
1 2 3 21 22 23 24 25 26 27 28 29 30 31 32 33
. . . .
. . . .
. . . .
. . . .
. . . .
. . . .
. . . .
.......
. . . . . . .
. . . . . . . . . . . . . . . . . . . . .
. . . . . . . 34 . 35 .
. . . . . . . . .
. . . . . . . . .
. . . . . . . . .
. . . . . . . . .
. . . . . . . . . . . . . . . . . .
36 . . . . . . . 37 . . . . . . . 38 . . . . . . . 39 . . . . . . .
40 . . . . . . . . . . . . .
41 . 42 . 43 . %. 45 .
. . . . . .
. . . . . . . . . . . . . . . . . .
chebulagicacid (35) geraniin(31) sanguiin H- 11 (26) 2.3.hexahydroxydiphenoyl. ~.glucose(26) strictinin( 17) pedunculagin (27) l(P)-O-galloylpedunculagin(28) eugeniin(29) corilagin (30) punicafolin (30) elmarpusin (32) furosin (33) terchebin (33) chebulinic acid (34) mallotinicacid (33) mallotusinic acid (33) sanguiin H-2 (26) punicalin (36) punicalagin (36) punicacorteinC(37) castalagin (38) vescalagin (38) grandinin (38. 39) casuarinin (38) phillyraeoidin A (40) sanguiin H-6 (26) castanopsininA (40) castanopsiniinA (40)
CondensedTannins
46 . 47 . 48 . 49 . 50 . 51 . 52 . 53 .
. . . . . . .
. . . . . . .
. . . . . . .
. . . . . . .
. . . . . . .
. . . . . . .
(+)-catechin (42) (-)-epicarechin (43) procyanidin B-2 (43) procyanidin C- 1 (43) (-)-epicatechin 3-0-gallate(42) procyanidinB.23.3I.di. O.gallate(42) procyanidinC.l3.3’.3”.tri. O.gallate(42) . . . . . . proanthocyanidin A-2 (44)
0.80 0.35 0.50 > 10 4.86 2.74 0.21 2.74 > 10 > 10 2.62 0.67 > 10 > 10 > 10 > 10 0.44 > 10 > 10 3.86 0.79 0.58 0.10 2.24 0.50 5 .00 > 10 > 10
> 10 > 10 > 10 > 10 0.55 3.45 3.05 5.50
1042
Journal of Natural Products
Compound
54 . . . . 55 . . . .
.. . .. .
Name cinnamtannin B, (43) cinnamtannin B, (43)
[Vol. 55, No. 8
ED,, (kg/ml) 3.57 0.66
Complex Tannins
56 . . . . . . . acutissimin A (38,45) 57
.. .....
mongolicainA(38,46) Actinomycin (positive control)
0.61
> 10 c0.01
MATERIALSAND METHODS MATERIhLS.--The 57 tannins and related compounds were isolated from the plant materials as reported previously (14-46) and shown in Schemes 1-3. BIOLOGICAL ASAY.--The in vitro cytotoxicity assay was carried out according to a National Cancer Institute protocol (47) as previously described (48). The assay was conducted using apanel ofhuman tumor cell lines. These cell lines are lung carcinoma (A-549), ileocecal adenocarcinoma(HCT-8), epidermoid carcinomaof nasopharnyx (KB), melanoma (RPMI-795 I), and medulloblastoma(TE-67 1). All cell lines were obtained from the American Type Culture Collection, Rockville, MD. In general, the assay methods were the same as those described by Monks et ul. (49). However, a tetrazolium salt method was used to measure cell survival and proliferation (50). ACKNOWLEDGMENTS This work was supported by grant from the National Cancer Institute No. CA-17625 (K.H. Lee).
LITERATURE CITED 1. K.H. Lee, J.X. Xie, and H. Hu, Abstracts ofchinere Medicine, 4 , 4 7 3 (1991). 2. T. Nagasawa, H. Oura, Y. Shoyama, and I. Nishioka, Chem. P h n n . Bull., 28, 1736 (1980). 3. S. Shibutani, T. Nagasawa, H. Oura, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 31,2378 (1983). 4. J. Inokuchi, H. Okabe, T. Yamauchi, A. Nagamatsu, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 33,264 (1985). 5. M.Nishizawa, T. Yamagishi, G.E. Dutschman, W.B. Parker, A.J. Bodner, R.E.Kilkuskie, Y.C. Cheng, and K.H. Lee,]. Nut. Prod., 52,762 (1989). 6. G. Nonaka, I. Nishioka, M. Nishizawa, T. Yamagishi, Y. Kashiwada, G.E. Dutschman, A.J. Bodner, R.E. Kilkuskie, Y.C. Cheng, and K.H. Lee,j.Nut. Prod., 53,587 (1990). 7. Y. Kimura, H. Okuda, T. Okuda, T. Hatano, I. Agata, and S.Archi, Plantu'Med.,50,473 (1984). 8. Y. Kimura, H. Okuda, T. Okuda, T. Hatano, I. Agata, and S. Archi, Chem. Phunn. BuII., 33, 2028 (1985). 9. T.Gichner, F. Posiisil,J. Veleminsky, V. Volkeova, and J. Volke, FoliuMirrobiol. (Prugue), 32,55 (1987). 10. S. Shirahata, H. Murakami, K. Nishiyama, I. Sugata, K. Shinohara, G. Nonaka, I. Nishioka, and H. Omura, Agrir. Biol. Chem., 49, 1033 (1985). 11. S. Shirahata, H . Murakami, K. Nishiyama, K. Yamada, G. Nonaka, I. Nishioka, and H. Omura, J . Agric. F w d C h . , 37,299 (1989). 12. S. Ueki, G. Nonaka, I. Nishioka, and M. Fujiwara, J . Med. Phunn. Sor. Wukan-YuLu, 2 , 502 (1985). 13. K. Miyamoro, N. Kishi, R. Koshiura, T. Yoshida, T. Hatano, and T.Okuda, Chem. Phunn. Bull., 35,814 (1987). 14. Y. Kashiwada, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 32,3461 (1984). 15. G. Nonakaand I. Nishioka, Chem. Phann. Bull., 31, 1652 (1983). 16. G. Nonaka, I. Nishioka, T.Nagasawa, and H. Oura, C h . Phunn. Bull., 29,2862 (1981). 17. G. Nonaka, R. Sakai, and I. Nishioka, Phytorhemistry, 23, 1753 (1984). 18. M. Nishizawa, T. Yamagishi, G. Nonaka, and I. Nishioka, J. Chem. Sor., Perkin Trans. 1 , 961 (1983).
August 19927 19. 20. 2 1. 22. 23.
24. 25. 26. 27. 28. 29. 30. 31. 32. 33. 34. 35. 36. 37. 38. 39. 40. 41. 42. 43. 44. 45. 46. 47. 48. 49.
50.
Kashiwada et al.: Tannins as Cytotoxic Agents
1043
G. Nonaka, I. Ishirnatsu, T. Tanaka, I. Nishioka, M. Nishizawa, and T. Yrnamagishi, Chem. Phunn. Bull., 35, 3127 (1987). M. Nishizawa, T. Yamagishi, G. Nonaka, and I. Nishioka,]. Chn. Soc., Perkin Trans. 1 , 2968 (1982). H. Nishimura, G. Nonaka, and I. Nishioka, Phytwhzstry, 23, 262 1 (1984). K. Ishirnaru, G. Nonaka, and I. Nishioka, P b y t o r h i m y , 25, 1501 (1987). M. Nishizawa, T. Yamagishi, G. Dutxhman, W.B. Parker, A.J. Bodner, R.E. Kilkuskie, Y.C. Cheng, and K.H. Lee,]. Nut. Prod., 5 2 , 762 (1989). G. Nonaka, M. Ageta, and I. Nishioka, Chnn. Phann. Bull., 36, 96 (1985). G. Nonaka, K. Ishirnaru, T. Tanaka, and I. Nishioka, Chem. Phum. Bull., 3 2 , 4 8 3 (1984). T. Tanaka, G. Nonaka, and I. Nishioka,]. C h . Res., Synop., 176 (1985);]. C h . Rer., Miniprint, 2001 (1985). H. Feng, G. Nonaka, and I. Nishioka, Pbytorhnnistry, 27, 1185 (1988). G. Nonaka, M. Ishimatsu, M. Ageta, and I. Nishioka, Chem. Phunn. Bull., 37, 50 (1989). G. Nonaka, M. Harada, and I. Nishioka, Chem. Phunn. Bull., 28, 685 (1980). T. Tanaka, G. Nonaka, and I. Nishioka, Phytwhemistry, 24, 2075 (1985). T. Tanaka, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 34, 941 (1986). T. Tanaka, G. Nonaka, I. Nishioka, K. Miyahara, and T. Kawasaki,]. Chem. Soc., Perkin Trans. I , 369 (1986). R. Saijo, G. Nonaka, and I. Nishioka, Chem. Pburm. Bull., 37, 2063 (1989). T.C. Lin, G. Nonaka, I. Nishioka, and F.C. Ho, Chem. Phunn. Bull., 38, 3004 (1990). T. Tanaka, G. Nonaka, andI. Nishioka, Chem. Pbunn. Bull., 34, 1039(1986). T. Tanaka, G. Nonaka, and I. Nishioka, Chem. Phann. Bull., 34,650 (1986). T. Tanaka, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 34,656 (1986). G. Nonaka, T. Sakai, T. Tanaka, K. Mihashi, and I. Nishioka, Chem. Pbunn. Bull., 38, 2151 (1990). G. Nonaka, K. Ishirnaru, R. Azuma, M. Ishirnatsu, and I. Nishioka, Chem. Phunn. Bull., 37,207 1 (1989). G. Nonaka, S. Nakayama, and I. Nishioka, Chem.Phunn. Bull., 37, 2030 (1989). M. Ageta, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 36, 1646 (1988). Y. Kashiwada, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 34,4083 (1986). S. Morimoto, G. Nonaka, and I. Nishioka, Chem. Pbunn. Bull., 34, 633 (1986). G. Nonaka, S. Morirnoto, and I. Nishioka,]. Chem. Soc., Perkin Truns. 1 , 2 139 (1983). K. Ishirnaru, G. Nonaka, and I. Nishioka, Chem. Phunn. Bull., 35, 602 (1987). G. Nonaka, K. Ishirnaru, K. Mihashi, Y. Iwase, M. Ageta, and I. Nishioka, Chem. Pbunn. Bull., 36, 857 (1988). R.I. Geran, N.H. Greenberg, N.M. McDonald, A.M. Schumacher, and B.J. Abbott, Cancer Chemotber. Rep., Part 3, 3, 1 (1972). K.H. Lee, Y.M. Lin, T.S. Wu, D.C. Zhang, T. Yamagishi, T. Hayashi, I.H. Hall, J.J. Chang, R.Y. Wu, and T.H. Yang, PIunta Med., 54, 308 (1988). A. Monks, D. Scudiero, P. Skehan, R. Shoemaker, K. Paull, D. Vistica, C. Hose, J. Langley, P. Cronise, A. Vaigro-Wolff, M. Gray-Goodrich, H. Campbell, J . Mayo, and M. Boyd,]. Nutl. Cunrerlnst., 83, 757 (1991). T. Mosrnann,J . fmmunol. Methodr, 65, 55 (1983).
Received 17 Ortober 1991
