J Nat Med (2010) 64:393401 DOI 10.

1007/s11418-010-0424-7

REVIEW

Search for bioactive natural products from medicinal plants of Bangladesh

Firoj Ahmed Samir Kumar Sadhu Masami Ishibashi

Received: 5 March 2010 / Accepted: 20 April 2010 / Published online: 6 July 2010 The Japanese Society of Pharmacognosy and Springer 2010

Abstract In our continuous search for bioactive natural products from natural resources, we explored medicinal plants of Bangladesh, targeting cancer-related tumor necrosis factor-related apoptosis-inducing ligand-signaling pathway, along with some other biological activities such as prostaglandin inhibitory activity, 1,1-diphenyl-2-picrylhydrazyl free-radical-scavenging activity, and cell growth inhibitory activity. Along with this, we describe a short eld study on Sundarbans mangrove forests, Bangladesh, in the review. Keywords Bangladesh Sundarbans mangrove forests TRAIL Apoptosis

medicinal plants, as does a large number of mangroves from Sundarbans mangrove forests. Medicinal plant resources of Sundarbans intrigued us greatly, so we studied the habitat and distribution patterns of mangroves growing there. Mangroves have long been a source of astonishment to scientists, including natural product researchers. This is because a diversied chemical class of compounds containing different types of biological activities has been isolated from mangroves [3]. In this report, we discuss the isolation and characterization of bioactive compounds from some medicinal plants of Bangladesh, along with a brief eld study on collection and species identication of mangroves from Sundarbans mangrove forests, Bangladesh.

Introduction Field study In search of bioactive natural products targeting cancerrelated signaling pathways such as tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), Wnt, and Hedgehog, we explored medicinal plant resources in Thailand and Bangladesh [1, 2]. Previously, we isolated a number of bioactive compounds targeting different biological activities such as prostaglandin (PG) inhibitory activity, 1,1-diphenyl-2-picrylhydrazyl (DPPH) free-radical-scavenging activity, and cell growth inhibitory activity. Bangladesh serves as a source of a wide variety of
F. Ahmed M. Ishibashi (&) Graduate School of Pharmaceutical Sciences, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan e-mail: mish@p.chiba-u.ac.jp F. Ahmed S. K. Sadhu Pharmacy Discipline, Life Science School, Khulna University, Khulna 9208, Bangladesh

Study area The eld study in Sundarbans mangrove forests was carried out in November 2008. Sundarbans is the largest single block of tidal halophytic mangrove forest in the world [4]. The Bengali language name Sundarban can be literally translated as beautiful jungle or beautiful forest (Sundar beautiful; ban forest or jungle). The name may have been derived from the Sundari trees (Heritiera fomes) that are found in Sundarbans in large numbers. The forest is spread across areas of Bangladesh and India and covers 10,000 km2, of which about 6,000 are in Bangladesh (Fig. 1a, b) [5]. It became inscribed as a United Nations Educational, Scientic, and Cultural Organization (UNESCO) World Heritage Site in 1997. Sundarbans is intersected by a complex network of tidal waterways, mudats, and small islands of salt-tolerant mangrove forests.

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394 Fig. 1 a Location of Sundarbans (http://news.bbc. co.uk); b map of Sundarbans (http://wwndia.org)

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Fig. 2 a Aerial roots (pneumatophores); b viviparous root, and c young seedlings of Rhizophora sp.; d trees on Sundarbans mainland

Sundarbans ora is characterized by the abundance of H. fomes, Excoecaria agallocha, Ceriops decandra, and Sonneratia apetala. A total 245 genera and 334 plant species were recorded in 1903 [6]. Study area included a number of spots in Sundarbans, such as Supati, Kochikhali, Katka, Dublar char, Nilkomol, and others. During our visit, we collected a number of species from Sundarbans. Salient features of mangroves Mangrove forests are composed of taxonomically diverse, salt-tolerant tree and other plant species, which thrive in intertidal zones of sheltered tropical shores, overwash islands, and estuaries. Mangrove trees have specially adapted aerial and salt-ltering roots and salt-excreting leaves that enable them to occupy the saline wetlands where other plant life cannot survive. They are present in tropical and subtropical areas around the world and are generally found within 25 north and south of the equator,

even though they can be found as high as 32 in some northern latitudes. Root system of mangroves is adapted to the peculiar conditions found in the mangrove forests, such as still roots in Rhizophora and knee roots in Bruguiera. Pneumatophores (breathing roots) are profuse in Sonneratia and Avicennia (Fig. 2a, d). Certain mangrove species can propagate successfully in a marine environment through viviparous germination, in which the seed germinates while still on the tree and falls during its germinating condition (Fig. 2b) [7]. Collection and identication Plants were collected from canal banks and deep forests in 2008 (Fig. 3). After collection, the plants were identied by Prof. AK Falul Huq, Forestry and Wood Technology Discipline, Khulna University, Bangladesh, who accompanied us on the entire eld study. In 2009, the study was arranged in Chittagong, and we collected a number of

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J Nat Med (2010) 64:393401 Fig. 3 a Plant collection of aerial parts from a shrub, b leaves with small stems from a tree, and c leaves with small stems from another tree; d people returning after collection

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medicinal plants at that time. Also, we plan to perform a similar study in some other parts of Bangladesh in 2010 and 2011. Chemical and biological studies on medicinal plants of Bangladesh During our search for bioactive natural products from medicinal plants of Bangladesh, we isolated a number of novel compounds and some known ones. In this section, we present the chemical and biological studies performed on some of the plants, such as Leucas aspera, S. caseolaris, Aphanamixis polystachya, Saraca asoca, Jasminum grandiorum, Zingiber spectabile, Hoya parasitica, Vallaris solanaceae, and Sida acuta. Leucas aspera L. aspera Link. (Labiatae), known as darkolos or dandokolos in Bangladesh, is a common aromatic herb and grows abundantly in Bangladesh and a wide area in south Asia. Traditionally, the decoction of the whole plant is taken orally for analgesicantipyretic, antirheumatic, anti-inammatory, and antibacterial treatment, etc., and its paste is applied topically to inamed areas. According to the traditional usage of the plant as an anti-inammatory and analgesic, L. aspera was tested for its PG inhibitory and antioxidant activities. The extract showed both activities, i.e., inhibition at 30 lg/ml against PGE1- and PGE2-induced contractions in guinea pig ileum and a DPPH-radical-scavenging effect. The separation guided by the activities in these dual assay

methods provided compounds 125, among which 17 and 1012 were identied as nectandrin B, meso-dihydroguaiaretic acid, macelignan, acacetin, apigenin 7-O[600 -O-(p-coumaroyl)-b-D-glucoside], chrysoeriol, apigenin, erythro-2-(4-allyl-2,6-dimethoxyphenoxy)-1-(4-hydroxy3-methoxyphenyl)propan-1-ol, myristargenol B, and machilin C, respectively (Fig. 4). Compound 8 was determined to be (-)-chicanine, the new antipode of the (?) compound, by spectroscopic methods, including circular dichroism (CD) and optical rotary dispersion (ORD). Chiral high-performance liquid chromatography (HPLC) analysis of compound 9 showed it was a mixture of two enantiomers: (7R,8R)- and (7S,8S)-licarin A [8, 9]. Sonneratia caseolaris S. caseolaris Linn. (Sonneratiaceae), locally known as Choila, is a small tree with oblong or obovate-elliptic coriaceous leaves and large red owers, which grows among mangrove areas ooded by the sea in the Sundarbans and Chittagong areas of Bangladesh. Extracts of this plant are traditionally used as an astringent and antiseptic, in sprains and swellings, and in arresting hemorrhage. Based on the traditional usage of this plant, we tested the extract for antioxidant activity using the DPPH-radicalscavenging effect on thin-layer chromatography (TLC). Although fatty acids, hydrocarbons, steroids, pectin, and sugars have been previously isolated from this plant [10], by taking the DPPH-radical-scavenging effect as the isolation guide, we isolated a avone, luteolin (26), and its 7-O-b-glucoside (cynaroside) (27) (Fig. 5). According to

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Fig. 6 Structures of compounds isolated from Saraca asoca (3239)

Fig. 4 Structures of compounds isolated from Leucas aspera (125)

plant of Bangladesh. The bark is astringent and is used to treat spleen and liver diseases, tumors, and abdominal complaints. Different types of compounds, for example, limonoids, terpenoids, glycosides, an alkaloid, and a saponin, have previously been isolated from this plant. With the objective of isolating components bioactive against tumors, a methanol (MeOH) extract of the dried bark, was checked for growth inhibitory activity against Henrietta Lacks (HeLa) cells and shown to have potent activity with an IC50 of approximately 50 lg/ml. The extract was then partitioned successively between hexane, ethyl acetate (EtOAc) and butanol (BuOH), and water. Of these, BuOH and n-hexane extracts were potent active fractions with IC50 of approximately 22 and about 33 lg/ ml, respectively. These two fractions were subjected to a general separation procedure to isolated compounds 2831, along with stigmasterol and oleic and linoleic acids (Fig. 5) [11]. Saraca asoca S. asoca (Roxb.) De Wilde or S. indica Linn. (family: Caesalpiniaceae; local names: Ashok, Anganapriya, etc.) is a medicinal plant of Bangladesh whose bark is astringent and used in menorrhagia, bleeding hemorrhoids, and hemorrhagic dysentery. The isolation of tannins, avonoids, proanthocyanidins, and leucoanthocyanidins were previously reported from the bark [12]. In our assay method, an MeOH extract of the dried bark showed potent antioxidant activity determined by DPPH-radical-scavenging assay. Following this activity, we isolated eight compounds (3239) from this plant (Fig. 6). Of the isolates, ve were lignan glycosides, such as, lyoniside, nudiposide, 5-methoxy-9-b-xylopyranosyl-(-)-isolariciresinol, icariside E3, and schizandriside; and three were avonoids, namely, (-)-epicatechin, epiafzelechin-(4b ? 8)-epicatechin, and procyanidin B2 [12].

Fig. 5 Structures of the compounds isolated from Sonneratia caseolaris (26 and 27) and Aphanamixis polystachya (2831)

the traditional medicinal usage of S. caseolaris, we tested the extract of S. caseolaris for antioxidant activity using DPPH-radical-scavenging effect with TLC. Following activity-oriented separation, two avonoids, luteolin (26) and luteolin 7-O-b-glucoside (27), were isolated. Both compounds were found to possess antioxidant activity [10]. Aphanamixis polystachya A. polystachya (Wall.) Parker or Amoora rohituka (family: Meliaceae; local names: Roina, Pitraj, etc.) is a medicinal

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Fig. 7 Structures of the compounds isolated from Jasminum grandiorum (4047) Fig. 8 Structures of the compounds isolated from Zingiber spectabile (4864)

Jasminum grandiorum J. grandiorum Linn. (family: Oleaceae; local names: Jasmine or Jatiful) is a medicinal plant of Bangladesh that has certain therapeutic properties against various psychiatric disorders, skin diseases such as conjunctivitis and dermatitis, and different types of cancer. Previously, iridoid-type compounds secoiridoid glucosides, triterpenes, avonoids, lignans, etc., were isolated from this herb [13]. In our continued investigation on traditional medicinal plants of Bangladesh, we isolated eight compounds (4047), including secoiridoid glucosides as 200 -epifraxamoside and demethyl-200 -epifraxamoside, the secoiridoid jasminanhydride (Fig. 7), together with four previously known phenolics and a triterpene from this herb. Structures were elucidated by detailed spectroscopic analysis. Stereochemistry of the compounds was determined by differential nuclear Overhauser effect (NOE) experiment [13]. Zingiber spectabile Z. spectabile (Zingiberaceae), common names: bonada (Bangla), beehive ginger (English), micro fono (Spanish), Wan-prai-dum (Thai), is a medicinal herb of Bangladesh primarily found in the Khagrachhari and Bandarban forests. Traditionally, its rhizome is used in cough and asthma complication and as a germicidal, stimulant, and tonic. It is reported that the rhizome extract is benecial in cancer. Previous investigations on the rhizome reported the presence of terpinen-4-ol, labda-8(17), 12-diene-15,16-dial, a-terpineol, a-pinene, b-pinene, limonene, and structurally related compounds [14]. Aiming at isolation of the components responsible for its anticancer property, we carried out a total phytochemical investigation guided by the cellgrowth inhibitory activity. Nine sesquiterpenes (4856) and eight avonoids (5764), together with b-sitosterol, were obtained, all of which were rst isolated from this species (Fig. 8) [14].

Fig. 9 Structures of the compounds isolated from Hoya parasitica (6570), Vallaris solanaceae (7173), and Sida acuta (7476)

Hoya parasitica H. parasitica Wall. ex Traill or Asclepias parasitica Wallich ex Hornemann (Asclepiadaceae, local name: Bayupriya, Porgacha) grows in southeastern Asia, particularly in the Sundarbans and Chittagong areas in Bangladesh. H. parasitica is a parasite creeper with a fragrant ower whose aerial part is used to treat rheumatism. A previous phytochemical investigation reported the isolation of dihydrocanaric acid, along with lupeol and lupenone, from this plant. However, no biological data have been reported. As part of our continuing investigation of traditional medicinal plants of southeastern Asia [3, 4], we isolated four compounds (6570) from this herb (Fig. 9). An androstanoid, hoyasterone (compound 65); a sesquiterpene, 15-bulnesolic acid (66); and a phenolic compound,

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1-(4-hydroxy-3-methoxyphenyl)-1-methoxypropan-2-ol (67); together with a known triterpene, dihydrocanaric acid (compound 68), were isolated from H. parasitica. Structures were elucidated by 1D and 2D nuclear magnetic resonance (NMR) and mass spectroscopic analysis [15]. Vallaris solanaceae V. solanaceae Kuntze (syn.: V. heynei) (Apocynaceae), locally known as agarmoni, grows in Bangladesh and in other Southeast Asian countries. It is a creeper with fragrant white owers traditionally used against ringworms and skin infections. Previous chemical investigation of its leaves reported its ability to isolate b-sitosterol, b-amyrin, ursolic acid, and triterpenes; vallaroside, solanoside, vallarosolanoside, and acoshimperosid P. In our continuous investigation on traditional medicinal plants of BangladeshAsia, we isolated a new cardenolide glycoside, vallarisoside (71), and a known one, 3b-O-(a-acofriosyl)16-anhydrogitoxigenin (72), along with a new glycoside, benzyl 2-O-b-apiofuranosyl-(1 ? 2)-b-D-glucopyranosyl2, 6-dihydroxy-benzoate (73) from this herb (Fig. 9) [16]. Sida acuta S. acuta Burm. (Malvaceae), locally known as Berela, is a shrub distributed in all areas of Bangladesh and other tropical countries. Its leaves are traditionally used as a diuretic, demulscent, etc. Roots are used as a tonic, diaphoretic, and antipyretic. It was reported for antiplasmodial, antimicrobial, analgesic, free-radical-scavenging, and apoptosisinducing activities. Previously isolated constituents include alkaloids, tocoferols, and triterpenoids; sterols poly phenols, and glycosides. Bioassay guided separation of Sida acuta whole plants led to the isolation of an alkaloid, cryptolepine (74); and two kaempferol glycosides (7576) (Fig. 9) [17]. The PG inhibitory activity The PG-inhibitory activity of isolates (125) was evaluated by the method using PG-induced contraction in guinea pig ileum. Compound 1 exhibited inhibition at 2.6 and 8.7 lM against PG E1- and PG E2-induced contractions, respectively, and 2 was inhibitory at 9.1 lM for both contractions. Compound 5 only caused inhibition at 5.2 lM against PG E1-induced contraction but was inactive against PG E2 at the same concentration. Compound 18 exhibited the most potent effect at 16 and 48 lM against PG E1- and PG E2-induced contractions, respectively. Compounds 13 and 17 inhibited both types of contractions at 126 and 76 lM, respectively, whereas 22 showed a less potent inhibition against PG E1-induced contraction only, at 98 lM [8, 9].

Antioxidant activity Antioxidant activities were evaluated by DPPH-radicalscavenging assay. In the case of antioxidant activity on TLC sprayed with DPPH reagent, all lignans and neolignans 13 and 812 showed positive spots on TLC, whereas the avonoids (47) were negative. The IC50 values of 13 and 5 and quercetin (positive control) recorded on a microplate reader with DPPH were 60, 28, 50, 500, and 30 lM, respectively. The IC50 values of DPPH-radical-scavenging assay for compounds 3239 and the positive control quercetin were 104, 85, 44, 75, 55, 50, 55, 40, and 30 lM, respectively [912]. Cell-growth inhibitory activity Compound 48 (zerumbone) was found to be the most active (IC50 59.6 lM) in cell-growth inhibitory assay against colon carcinoma SW480 cells. Among the isolated compounds from H. parasitica, only dihydrocanaric acid (68) exhibited growth inhibitory activity against both HeLa and SW480 cells [14, 15]. TRAIL-resistance-overcoming activity Tumor-necrosis-factor-related apoptosis-inducing ligand (TRAIL), a member of the TNF superfamily, has emerged as a promising anticancer agent because of its ability to selectively kill tumor cells. TRAIL-induced apoptosis initiated by the death-receptor pathway involves DR engagement, death-inducing signaling complex (DISC) formation, proteolytic activation of caspase-8, and, consequently, activation of caspase-3. Proteolytic caspase-8 further activates Bid, which, in turn, translocates to the mitochondria and activates the mitochondrial pathway. However, considerable numbers of cancer cells, especially some highly malignant tumors, are resistant to apoptosis induction by TRAIL. Resistance to TRAIL can occur at different points in the signaling pathways of TRAILinduced apoptosis. Overcoming TRAIL resistance and understanding the mechanisms underlying such resistance are thus very important in anticancer drug discovery [1]. Isolated compounds (7176) were evaluated for their activity in overcoming TRAIL resistance in human gastric adenocarcinoma (AGS) cells. Recently, this cell line has been widely used as a model system for evaluating cancer cell apoptosis [16] and is reported to be refractory to apoptosis induction by TRAIL. To assess the effects of the isolated compounds, TRAIL, or their combined treatment on cell viability, AGS cells were treated with the indicated agents and subjected to uorometric microculture cytotoxicity assay (FMCA) [18, 19]. As shown in Fig. 10a, treatment with 100 ng/ml TRAIL for 24 h resulted in only

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Fig. 10 a Effect of compound 71, luteolin (positive control: Lut) and tumor-necrosis-factor-related apoptosis-inducing ligand (TRAIL) treatment, alone and in combination, on the viability of human gastric adenocarcinoma (AGS) cells. Cells were seeded in a 96-well culture plate (6 9 103 cells per well) for 24 h and then treated with indicated concentrations (lM) of the compounds and/or TRAIL for 24 h. Cell viability was determined by uorometric microculture cytotoxicity assay (FMCA). The bar represents the means [n = 3 standard deviation (SD)]. Signicance was determined by Students t test; p \ 0.01 (**) vs. control (Con). b Effect of compound 74, luteolin (positive control: Lut) and TRAIL treatment, alone and in combination, on the viability of AGS cells. Cells were seeded in a 96-well culture plate (6 9 103 cells per well) for 24 h and then treated with indicated concentrations (lM) of the compounds and/or TRAIL for 24 h. Cell viability was determined by uorometric microculture cytotoxicity assay (FMCA). The bar represents the means (n = 3 SD). Signicance was determined by Students t test p \ 0.01 (**) vs. control (Con)

a slight decrease in cell viability (89%). Luteolin [20, 21], used as a positive control, produced about 50% more inhibition along with TRAIL than the agent alone. Combined treatment with TRAIL and compound 71 at 5, 10, and 20 lM produced about 32%, 37%, and 31% more inhibition than the agent alone. These results suggest a possible synergism between compound 71 and TRAIL. Combined treatment of AGS cells with 100 ng/ml TRAIL and 1.25 or 2.5 lM of compound 74 produced about 32% and 50% more inhibition, respectively, than the agent alone, suggesting a possible synergism between the two agents. Cryptolepine (compound 74), reported as a candidate antitumor agent [22, 23], exhibits potent cytotoxic activity against a wide variety of cancer cells, including human leukemia HL-60 cells [24]. It induced cell-cycle arrest and apoptosis by activating mitochondrial release of cytochrome c. Here, we found, for the rst time, its TRAIL-resistance-overcoming activity against AGS cells (Fig. 10b). To ascertain whether the decrease in cell viability produced by compound 74 was caused by apoptotic cell death, we stained the treated cells after 24 h with Hoechst 33342 reagent. We observed apoptotic nuclei, with condensed chromatins stained more brightly in the treated cells than the normal cells (Fig. 11), suggesting that the cell death was due to apoptosis [25]. We further checked the effect of compound 74 on caspase-3/7 activity in AGS cells to ascertain whether the induced apoptosis was mediated through caspase activation. Caspases-3/7 are known as effector caspases, and after activated by the initiator caspases (caspase-8/9), it induce apoptosis. We observed that treating AGS cells with compound 74 in combination with TRAIL (100 ng/ml), increased caspase-3/7 activity 1.8-, 1.9-, and 2.3-fold, at 1.25, 2.5, and 5 lM, respectively, compared with the control after 12 h (Fig. 12). The results tend to suggest that compound 74 sensitized AGS cells to TRAIL-induced apoptosis through caspase-3/7 activation [26].

Fig. 11 Effect of compound 74 and tumor-necrosis-factor-related apoptosis-inducing ligand (TRAIL) treatment, alone and in combination, on apoptosis of human gastric adenocarcinoma cells (AGA). AGS cells were grown on cell culture dishes and treated as described,

and apoptosis was detected by Hoechst 33342 stain. Representative photomicrographs from each treatment group showing induction of apoptosis (bright uorescence indicated by arrow)

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J Nat Med (2010) 64:393401 3. Bandaranayake WM (2002) Bioactivities, bioactive compounds and chemical constituents of mangrove plants. Wetl Ecol Manag 10:421452 4. Pasha MK, Siddiqui NA (2003) Sundarbans. In: Islam S (ed) Banglapedia: national encyclopedia of Bangladesh. Asiatic Society of Bangladesh, Dhaka 5. Sanderson E, Forrest J, Loucks C, Ginsberg J, Dinerstein E, Seidensticker J, Leimgruber P, Songer M, Heydlauff A, OBrien T, Bryja G, Klenzendorf S, Wikramanayake E (2006) Setting priorities for the conservation and recovery of wild tigers: 2005 2015. The Technical Assessment. WCS, WWF, Smithsonian, NFWF-STF, Washington, DC 6. Prain D (1903) The ora of Sundarbans. Rec Bot Surv India 114:231272 7. Maltby E (1986) Waterlogged wealth: why waste the worlds wet places?. International Institute for Environment and Development, London, p 200 8. Sadhu SK, Okuyama E, Fujimoto H, Ishibashi M (2003) Separation of Leucas aspera, a medicinal plant of Bangladesh, guided by prostaglandin inhibitory and antioxidant activities. Chem Pharm Bull 51:595598 9. Sadhu SK, Okuyama E, Fujimoto H, Ishibashi M (2006) Diterpenes from Leucas aspera inhibiting prostaglandin-induced contractions. J Nat Prod 69:988994 10. Sadhu SK, Ahmed F, Ohtsuki T, Ishibashi M (2006) Flavonoids from Sonneratia caseolaris. J Nat Med 60:264265 11. Sadhu SK, Phattanawasin P, Choudhuri MSK, Ohtsuki T, Ishibashi M (2006) A new lignan from Aphanamixis polystachya. J Nat Med 60:258260 12. Sadhu SK, Khan MS, Ohtsuki T, Ishibashi M (2007) Secoiridoid components from Jasminum grandiorum. Phytochemistry 68:17181721 13. Sadhu SK, Khatun A, Ohtsuki T, Ishibashi M (2007) First isolation of sesquiterpenes and avonoids from Zingiber spectabile and identication of zerumbone as the major cell growth inhibitory component. Nat Prod Res 21:12421247 14. Sadhu SK, Khatun A, Phattanawasin P, Ohtsuki T, Ishibashi M (2007) Lignan glycosides and avonoids from Saraca asoca with antioxidant activity. J Nat Med 61:480482 15. Sadhu SK, Khatun A, Ohtsuki T, Ishibashi M (2008) Constituents from Hoya parasitica and their cell growth inhibitory activity. Planta Med 74:760763 16. Ahmed F, Sadhu SK, Ohtsuki T, Khatun A, Ishibashi M (2010) Glycosides from Vallaris solanaceae with TRAIL-resistanceovercoming activity. Heterocycles 80:477488 17. Ahmed F, Toume K, Ohtsuki T, Rahman M, Sadhu SK, Ishibashi M (2010) Cryptolepine, isolated from Sida acuta, sensitizes human gastric adenocarcinoma cells to TRAIL-induced apoptosis (in press) 18. Larsson R, Kristensen J, Sandberg C, Nygren P (1992) Laboratory determination of chemotherapeutic drug resistance in tumor cells from patients with leukemia, using a uorometric microculture cytotoxicity assay (FMCA). Int J Cancer 50:177185 19. Ahmed F, Ohtsuki T, Aida W, Ishibashi M (2008) Tyrosine derivatives Isolated from Streptomyces sp. IFM 10937 in a screening program for TRAIL-resistance-overcoming activity. J Nat Prod 71:19631966 20. Horinaka M, Yoshida T, Shiraishi T, Nakata S, Wakada M, Nakanishi R, Nishino H, Matsui H, Sakai T (2005) Luteolin induces apoptosis via death receptor 5 up-regulation in human malignant tumor cells. Oncogene 24:71807189 21. Horinaka M, Yoshida T, Shiraishi T, Nakata S, Wakada M, Nakanishi R, Nishino H, Sakai T (2005) The combination of TRAIL and luteolin enhances apoptosis in human cervical cancer HeLa cells. Biochem Biophys Res Commun 333:833838

Fig. 12 Effect of compound 74 on caspase-3/7 activity in human gastric adenocarcinoma (AGS) cells. Effect of combined treatment of compound 74 and tumor-necrosis-factor-related apoptosis-inducing ligand (TRAIL) on caspase-3/7 activity in human gastric adenocarcinoma cells. Values on top of the columns represent relative fold induction compared with control, considered as 1. Signicance was determined by Students t test (**p \ 0.01; *p \ 0.01) vs. control (Con)

Conclusion Mangrove plants may serve as a potential source of bioactive compounds, but little is known about the chemistry of natural products of mangroves. We studied the habitat and distribution of mangroves and collected some species for further exploration for isolation and characterization of bioactive compounds from them. In this report, we also describe our recent ndings on our search for bioactive natural products from some medicinal plants of Bangladesh, targeting different biological activities, as, for example, TRAIL-resistance-overcoming activity, PG inhibitory activity, DPPH-free-radical-scavenging activity, and cell-growth inhibitory activity. We were able to isolate a number of compounds with signicant activity.
Acknowledgments We are grateful to Prof. AK Falul Huq, Forestry and Wood Technology Discipline, Khulna University, Bangladesh, for his kind effort to guide us throughout the study and identication of the mangrove plants. We are also grateful to the Tokyo Biochemical Research Foundation (TBRF) who provided Post Doctoral Fellowship to Dr. Samir K. Sadhu. This work was supported by the Grant-in-Aid for Scientic Research from the Japan Society for the Promotion of Science (JSPS).

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