Phytochemical Analysis and Anticancer Properties of Drimia maritima Bulb Extracts on Colorectal Cancer Cells.

Cancer is a worldwide health problem and is the second leading cause of death after heart disease. Due to the high cost and severe side effects associated with chemotherapy treatments, natural products with anticancer therapeutic potential may play a promising role in anticancer therapy. The purpose of this study was to investigate the cytotoxic and apoptotic characteristics of the aqueous Drimia maritima bulb extract on Caco-2 and COLO-205 colorectal cancer cells. In order to reach such a purpose, the chemical composition was examined using the GC-MS method, and the selective antiproliferative effect was determined in colon cancer cell lines in normal gingival fibroblasts. The intracellular ROS, mitochondrial membrane potential, and gene expression changes in selected genes (CASP8, TNF-α, and IL-6 genes) were assessed to determine the molecular mechanism of the antitumor effect of the extract. GC-MS results revealed the presence of fifty-seven compounds, and Proscillaridin A was the predominant secondary metabolite in the extract. The IC50 of D. maritima bulb extract on Caco-2, COLO-205, and the normal human gingival fibroblasts were obtained at 0.9 µg/mL, 2.3 µg/mL, and 13.1 µg/mL, respectively. The apoptotic effect assay indicated that the bulb extract induced apoptosis in both colon cancer cell lines. D. maritima bulb extract was only able to induce statistically significant ROS levels in COLO-205 cells in a dose-dependent manner. The mitochondrial membrane potential (MMP) revealed a significant decrease in the MMP of Caco-2 and COLO-205 to various concentrations of the bulb extract. At the molecular level, RT-qPCR was used to assess gene expression of CASP8, TNF-α, and IL-6 genes in Caco-2 and COLO-205 cancer cells. The results showed that the expression of pro-inflammatory genes TNF-α and IL-6 were upregulated. The apoptotic initiator gene CASP8 was also upregulated in the Caco-2 cell line and did not reach significance in COLO-205 cells. These results lead to the conclusion that D. maritima extract induced cell death in both cell lines and may have the potential to be used in CRC therapy in the future.

Beneficial properties of Drimia numidica leaf methanolic extract against the cytogenotoxic effects of mitomycin C on human lymphocytes.

This study investigated the phytochemical profile of Drimia numidica leaf methanolic extract, as well as its cyto-genotoxic and cyto/genoprotective potential against mitomycin C (MMC) mediated effects on healthy human lymphocytes. Photosynthetic pigments, trace elements, and secondary metabolites were estimated and/or identified in methanolic extract of mature leaves, and the latter was further used for assessing its in vitro biological effects on MMC-free and/or MMC-treated human lymphocytes (at low, non-toxic concentrations of 0.001 and 0.01% v/v). The results showed that D. numidica leaf methanolic extract, being rich in carotenoids, phenolics, flavonoids, organic acids and bufadienolides, could be protective against MMC mediated cyto/genotoxic potential in healthy human lymphocytes. Biomolecules possessing antioxidant and antitumor potential, such as beta-carotene and lutein among others, chlorogenic acid, caffeic acid and their derivatives, minerals such as Si, as well as apigenin- and luteolin-derived glycosides, either individual or in a mixture, could be beneficial rather than harmful, at least at the extract concentrations tested. Although further in vitro and in vivo studies are still needed for elucidating the beneficial (individual and/or additive/synergistic) role of those compounds, the results of the present study are quite promising, thus encouraging new challenges for the appropriate utilization of D. numidica leaf extract.

Antiproliferative Bufadienolides from the Bulbs of Drimia altissima.

The bulbs of the South African Drimia altissima (Asparagaceae or Hyacinthaceae sensu APGII) have yielded a range of previously undescribed bufadienolides, drimianins A-G (1-7), the known bufadienolides bovogenin A (8), 3ß-O-ß-d-glucopyranosylbovogenin A (9), scillaren F (10), and altoside (11), the known homoisoflavonoid (3S)-3-(4'-methoxybenzyl)-5,6,7-trimethoxychroman-4-one (urgineanin C), the sesquiterpenoids 1ß,6α-dihydroxy-4(15)-eudesmene and 6α-hydroxy-4(15)-eudesmen-1-one, polybotrin, adenosine, and 9R-hydroxy-(10E,12Z)-octadecadienoic acid ethyl ester. The bufadienolides isolated were tested at 10 µM in the NCI-60 cancer cell screen, and nine of these were selected for further screening at five concentrations. Drimianins C (3) and E (5) showed activity at the nanomolar level against a number of human cancer cell lines in the NCI-60 screen.

Isolation and characterisation of altissimin: a novel cytotoxic flavonoid C-apioglucoside from Drimia altissima (Asparagaceae).

Flavonoids are a class of biologically active compounds with various proven nutraceutical benefits. In flavonoid C-glycosides, the aglycones are attached to sugar residues via cleavage-resistant C-C bonds which alter typical flavonoid pharmacokinetic properties. In these compounds, the combination of biological activities from the flavonoid moieties and sugar residues create unique and more diverse biological functions than those of O-glycosylated and unsubstituted flavonoids. Through a series of reverse phase chromatography techniques and various spectroscopic methods, the phytochemical investigation of Drimia altissima (L.F.) Ker Gawl., a specie from the Asparagaceae family, led to the isolation and chemical characterisation of a novel C-glucosylflavonoid, altissimin, with a unique apioglucoside arrangement to the apigenin aglycone. Altissimin was found to possess strong in vitro anti-proliferative activity against HeLa cervical cancer cells.

Therapeutic Aspects of Squill; An Evidence-Based Review.

From ancient times, medicinal plants have been usually utilized to treat many disorders, but today, interest in these herbs is again aroused, because of their fewer side effects and low-cost. In traditional medicine, for many diseases, various medicinal herbs have been suggested so far. Drimia maritime, also named squill, is an important medicinal plant for the treatment of many diseases, especially respiratory diseases. In the current evidence-based study, we conducted a review of the general characteristics, ingredients, administration form, and side effects of squill in traditional medicine. For this purpose, traditional Persian medicine literatures and electronic databases were examined including PubMed, Scopus, and Google Scholar. Many compounds are isolated from D.maritima, including scillaren, scillirubroside, scillarenin, and bufadienolide glycosides. Oxymel is the most commonly used form of squill for various diseases, especially respiratory diseases. Besides, squill has been used in the treatment of cardiovascular, digestive, and dermatological disorders, it is also used against various cancer cells for its antioxidant and cytotoxic properties. Moreover, there is relatively reliable evidence of its benefits for bacterial and helminthic infections, rheumatism, edema, gout, abortion induction, healing of wounds and urine induction. It seems that supplementary studies are required to explore the bioactive agents and their effective mechanisms.

Characterization of the chemical composition of Drimia numidica plant parts using high-resolution mass spectrometry: study of their total phenolic content and antioxidant activity.

Drimia species have been used since ancient times for their medicinal properties. Their bulbs are considered as the main source of secondary metabolites with biological activity but the chemical composition of the other plant parts has not yet been considered. The aim of this study is to contribute to the existing knowledge with new data on the total phenolic content, the antioxidant activity and the chemical profile of different parts of Drimia numidica. The total phenolic content was estimated by the Folin-Ciocalteu assay and the antioxidant activity with DPPH· and ABTS·+ reagents. The separation and the identification of the compounds were performed with liquid chromatography combined with time-of-flight high-resolution mass spectrometry (LC/Q-TOF/HRMS). The extract of leaves presented the highest phenolic content while the highest antioxidant activity was presented by the extract of flowers. Results of the chemical analysis verify the presence of bufadienolides and phenolic compounds. Graphical abstract.

Chemical characterization and acaricidal activity of Drimia maritima (L) bulbs and Dittrichia viscosa leaves against Dermanyssus gallinae.

The emergence of resistance to chemical acaricides in Dermanyssus gallinae, together with their toxicity and high costs, has prompted investigations into the use of plant extracts as alternatives to chemical acaricidal treatments. Drimia maritima bulbs and Dittrichia viscosa (D. viscosa) leaf extracts were here characterized by HPLC-PDA-ESI-MS/MS, and their toxicity against D. gallinae was evaluated using contact methods. Twenty-nine compounds were identified in D. maritima extracts, with glucoscilliphaeoside derivatives (i.e., quercetin, kaempferol and bufadienolides) as the major components. Twenty-four phenolic compounds, mainly caffeic acid derivatives, were detected in D. viscosa extracts. D. maritima extracts displayed a significantly higher (p < 0.05) acaricidal activity than D. viscosa extracts, with 100% of D. gallinae mortality at a concentration of 100 mg/mL following 24 h exposure. The mortality rate of D. gallinae induced by D. viscosa extracts ranged from 25 to 45% following 48 h exposure at a concentration of 200 mg/mL. The acetonic extract of D. viscosa and D. maritima displayed the highest efficacy against D. gallinae. This study provides evidence of the diversity of bioactive compounds present in D. maritima bulbs and D. viscosa leaf extracts, which are both efficacious against D. gallinae. The higher efficacy of D. maritima bulb extracts might be linked to the presence of bufadienolides in its extracts.

Biochemical profile of non-enzymatic stress markers in the plant species "Urginea maritima" in a Mediterranean natural reserve exposed to oxidative stress.

Protected areas decrease degrading natural ecosystems due to pollution such as air pollution. In 1981, the inhabitants founded Bentael natural reserve in Byblos, Lebanon, to secure their region against urbanization projects, like the recently constructed road that threatens the biodiversity of the reserve. This study was conducted to determine the oxidative stress resulting from this pollution and that menaces 360 floral species among them a rare species "Urginea maritima." In this research, the biomonitoring approach was experienced to assess the oxidative stress. Biomonitoring possesses has the advantage to be low cost and a constructive method to generate valuable data for further examinations. The studied parameters were air pollutants, ascorbic acid, photosynthetic pigments, leave's pH, relative water content, proline, carbohydrates, and hydrogen peroxide, in three chosen spots, near the pollution source (P1), opposite the latter spot (P2), and in an area relatively far from the source of contamination and which was chosen as the control site (Ctrl). The results showed in P1 detection of air pollutants higher of about 80% than in Ctrl, modifications in stress markers: increased concentration of the reactive oxygen species "hydrogen peroxide," rise in the concentration of the osmoregulator amino acid "proline," and depletion in chlorophyll content, in contrast to an increase in pheophytin. All these findings can be exploited as early diagnosis of air pollution and confirmed the ability to use such biomonitor ("Urginea maritima") as a way to assess the environmental pollution levels and consequently affirm the danger of such landscape activities on natural reserves.

C-glycosyl flavone from Urginea indica inhibits proliferation & angiogenesis & induces apoptosis via cyclin-dependent kinase 6 in human breast, hepatic & colon cancer cell lines.

BACKGROUND & OBJECTIVES: Search for novel compounds beneficial to the treatment of cancer attracts a great deal of attention. We earlier demonstrated the isolation of 5,7-dihydroxy-2-[4'-hydroxy-3'-(methoxymethyl)phenyl]-6-C-ß-glucopyranosyl flavone, a novel C-glycosyl flavone from Urginea indica bulb. The present study was undertaken to investigate the effect of this novel compound on human normal epithelial and breast, hepatic and colon cancer cell lines. METHODS: : The maximum non-toxic concentration (MNTC) and cytotoxicity of C-glycosyl flavone were assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Cell cycle was analyzed by flow cytometry. Docking studies were performed to predict possible targets. Levels of cyclin-dependent kinase 1 (CDK1) and CDK6, Bcl2 and BAX and cytochrome c were quantified by specific ELISA. Mitochondrial membrane potential was determined using JC-1 dye. Apoptosis was quantified by Annexin V ELISA method. RESULTS: : Flow cytometry analysis demonstrated G0/G1 arrest. In silico docking studies predicted CDK1 and CDK6 as a possible target of C-glycosyl flavone. In vitro study confirmed CDK6 as the main target in C-glycosyl flavone-treated cancer cell lines. C-glycosyl flavone treatment also induced membrane blebbing, chromatin fragmentation and nucleosome formation. C-glycosyl flavone treatment caused marked loss of mitochondrial membrane potential, decrease in Bcl2/BAX ratio and activation of caspase-3 and release of caspase-9 and cytochrome c. In addition, C-glycosyl flavone inhibited the tumour-induced angiogenesis and reduced the vascular endothelial growth factor levels. Similarly, CDK6 inhibitor significantly inhibited proliferation and angiogenesis and induced apoptosis in tested cell lines. INTERPRETATION & CONCLUSIONS: The results indicate that C-glycosyl flavone may exert induction of apoptosis, cell cycle arrest and inhibition of angiogenesis via CDK6. Thus, targeting CDK6 using C-glycosyl flavone may serve as a novel therapeutic approach for the treatment of breast, hepatic and colon cancers.

Chromatographic fractionation, antioxidant and antibacterial activities of Urginea maritima methanolic extract.

The present work concerns a phytochemical study of Urginea maritima L. from Algeria, and an evaluation of antioxidant activity of the methanolic extract (UMME) and its chromatographic fractions. UMME was fractionated using open glass chromatography on silica gel and antioxidant effects were evaluated using DPPH and ß-carotene/linoleate assays. The phytochemical screening revealed that the bulb of plant contains flavonoids, glycosides, tannins, reducing compounds, anthraquinones combined, anthocyanins, mucilage, triterpenes and steroids. DPPH method showed that the UMME has a scavenger effect on radical DPPH with an IC50=57.83±1.59µg/ml. The fractions isolated from U. maritima (L.) presented an IC50 ranging between 499.23 and 39.68µg/ml. In ß-carotene/linoleate test, UMME and fractions give an I% =69.56±0.08% and between 31.29±0.49% and 90.79±0.29%, respectively. UMME showed a high inhibitory effect on the xanthine oxidase (IC50=0.67±0.01 mg/ml) and on the cytochrome c reduction (IC50=0.68 mg/ml). Wide range of phytochemical constituents in Urginea maritima were detected in methanolic extract which exhibited antioxidant and antibacterial activity. This plant could serve as pilot for the development of novel agents for pathological disorders.

C-glycosyl Flavone from Urginea indica Inhibits Growth and Dissemination of Ehrlich Ascites Carcinoma Cells in Mice.

BACKGROUND: C-glycosyl flavone, a phytochemical constituent in U.indica bulb, has been reported to possess cytotoxic activity. OBJECTIVE: The present study aims to investigate the toxicity and anticancer potentials of C-glycosyl flavone against Ehrlich ascites carcinoma mice model. METHOD: In present study, acute and chronic toxicity along with antitumor activity of C-glycosyl flavone isolated from U.indica bulb were Performed using in vitro and in vivo methods. Acute and chronic toxicity of C-glycosyl flavone was evaluated using Swiss albino mice. The effect of C-glycosyl flavone on proliferation of Ehrlich ascites carcinoma (EAC) cells was determined. Further, growth inhibition and dissemination were studied using EAC induced mice model. RESULTS: C-glycosyl flavone showed significant therapeutic potency against EAC cells in terms of reduced viability, cell cycle arrest, induction of apoptosis, inhibition of capillary formation, reduced VEGF levels. Moreover, there was reduction in body weight, tumor volume, viable tumor cells, increased survival of EAC induced mice upon C-glycosyl flavone treatment. Treatment also reduced dissemination of EAC cells into heart, kidney, liver and brain and diminished the pathological alterations induced by EAC cells in mice. In addition, there was an improvement in hemoglobin levels and counts of RBC, neutrophils, lymphocytes and monocytes in C-glycosyl flavone-treated mice with tumor. An enhancement of antioxidant status in C-glycosyl flavone treated EAC-bearing mice which appeared in terms of decreased serum thiobarbituric acid reactive substance and lipid peroxidation, increased GSH, SOD, Catalase and GPX. These results were comparable to a standard 5- fluorouracil treatment. C-glycosyl flavone exhibited safety profile in toxicity studies. CONCLUSION: Our study confirms the therapeutic potency of C-glycosyl flavone against EAC in inhibition of dissemination and growth of EAC in mice.

Traditional medical uses of Drimia species in terms of phytochemistry, pharmacology and toxicology.

Drimia genus includes plants that used from ancient time for various ailments such as dropsy, respiratory ailment, bone and joint complications, skin disorders, epilepsy and cancer. Toxic properties of some Drimia species also were noted by ancient scientists and these plants have been traditionally used for rat control. Bufadienolides have been identified as the main constituents in the genus of Drimia. Phenolics, sterols, protein and some of other phytochemicals have been also isolated from these plants. Pharmacological and clinical studies have strongly approved their effect on cardiovascular system. Extracts and compounds isolated from Drimia species showed biological activities such as antibacterial, antifungal, antiviral, antioxidant, anti-inflammatory and insecticidal effects through several in vivo and in vitro studies. Moreover, cytotoxic and antitumor activities which may be related to bufadienolide content of these plants have been considered by many researchers. Traditional therapeutic values of these plants for treating respiratory and rheumatic ailments as well as skin disorders are needed to be validated through more researches. Toxic effects of these plants and isolated compounds have been investigated through several in vivo studies. Drimia plants and their isolated compounds have narrow therapeutic index, so patients should be prohibited from applying these plants without medical supervision and should be informed about the main intoxication symptoms before starting treatment. Moreover, interaction of Drimia plants with other constituents of traditional herbal mixtures as well as chemical and biological modalities for reducing toxicity of bufadienolide compounds can be subjected for future studies.

New alkylresorcinols from a lipophilic extract of Urginea indica L. bulbs showing experimental trauma healing activity.

Several alkylresorcinols presenting the substitution pattern of structures I (3-methyl ether of 5-alkyl-2-methylresorcinol) and II (1,3-dimethyl ether of 5-alkylresorcinol), were isolated from the dichloromethane extract of the air-dried bulbs of Urginea indica L. Compounds of structure I with 15, 17 and 20 carbon atoms in the alkyl chain as well as compounds of structure II with 20, 22, 24 carbon atoms in the alkyl chain are new. The structures of the new compounds were elucidated on the basis of their NMR and MS data. The exact number of homologues in each series I and II and the exact length of the side chain were found using GC-MS analysis. The dichloromethane extract of the bulbs was evaluated for its trauma healing properties after local application and a statistically significant tendency to trauma remodeling was observed in comparison to control groups.

Antiproliferative Homoisoflavonoids and Bufatrienolides from Urginea depressa.

Investigation of the South African plant Urginea depressa Baker (Asparagaceae Juss.) for antiproliferative activity against the A2780 ovarian cancer cell line led to the isolation of the six new homoisoflavonoids urgineanins A-F (1-6), the two known bufatrienolides 7 and 9, and the new bufatrienolides urginins B and C (8 and 10). Structures were elucidated based on analysis of their 1D and 2D NMR spectra, electronic circular dichroism, and mass spectrometric data. Five of the six new homoisoflavonoids had good antiproliferative activity against the A2780 ovarian cancer, A2058 melanoma, and H522-T1 human non-small-cell lung cancer cells, and urgineanin A (1) had submicromolar activity against all three cell lines. The four bufatrienolides 7-10 had strong antiproliferative activity against the same cell line, with IC50 values of 24.1, 11.2, 111, and 40.6 nM, respectively.

Gastrointestinal stimulant effect of Urginea indica Kunth. and involvement of muscarinic receptors.

Urginea indica Kunth. (Family; Liliaceae) was studied for its gastrointestinal stimulant effect to rationalize the traditional medicinal uses as a digestive aid, stomachic and laxative. The crude aqueous-methanol extract of Urginea indica bulb (Ui.Cr) was tested on mice and isolated gut preparations. Ui.Cr, which was tested positive for alkaloids, tannins and coumarins, increased faecal output and accelerated charcoal meal transit in mice (6-12 mg/kg, p.o.), similar to that caused by carbachol (10 mg/kg). Ui.Cr (0.01-1 mg/mL) caused a spasmogenic effect in guinea-pig ileum that was reproduced in rabbit jejunum (0.01-0.3 mg/mL) followed by relaxation at a higher concentration. Like carbachol, the stimulant effect of Ui.Cr was blocked by atropine, suggesting the activation of muscarinic receptors mediating the prokinetic effect. Ui.Cr (0.01-5.0 mg/mL) also inhibited K(+) (80 mm)-induced contraction in rabbit jejunum and shifted the Ca(2+) concentration-response curves to the right, similar to verapamil, a standard calcium channel blocker. These data, indicating the presence of a gastrointestinal stimulant effect in Urginea indica possibly mediated through a cholinergic mechanism, provide a rationale for the use of Urginea indica in indigestion and constipation. The presence of a calcium antagonist effect in the plant may help to alleviate untoward effects of the plant that may result from an excessive increase in gut motility.

Novel flavonoid glycosides from the bulbs of Urginea indica Kunth.

Three novel flavonoid glycosides, 5,6-dimethyoxy-3',4''-dioxymethylene-7-O-(6''-beta-D-glucopyranosyl-beta-D-glucopyranosyl) flavanone (1), 5,4'-dihydroxy-3-O-alpha-L-rhamnopyranosyl-6-C-glucopyranosyl-7-O-(6''-para-coumaroyl-beta-D-glucopyranosyl) flavone (2) and 5,4'-dihydroxy-3-O-(2'''''-beta-glucopyranosyl-alpha-L-rhamnopyranosyl)-6-C-glucopyranosyl-7-O-(6''-para-coumaroyl-beta-D-glucopyranosyl) flavone (3) were isolated from the 1-butanol soluble fraction of the bulbs of the plant Urginea indica (Indian squill). The structures of the compounds were elucidated on the basis of spectral analysis, including homo- and heteronuclear correlation NMR experiments (COSY, NOESY, HSQC and HMBC) and mass spectra.

Non-target effect of organic insecticides: effect of two plant extracts on soil microbial biomass and enzymatic activities in soil.

Efficacious botanical derivatives can provide an alternative to synthetic pesticides for organic farming systems. However, there is lack of information regarding the side effects of organic pesticides on key soil ecological processes. In this study, we investigated the effects of aqueous extracts from Urginea maritima and Euphorbia myrsinites exhibiting translaminar and systemic activity against pests on microbial biomass and enzymatic activities in soil. Two grams of plant material was extracted with 100 ml of water and then diluted 1:100, 2:100, and 4:100 with distilled water. Diluted plant extracts were applied around hypocotyl of tomato by soil drench. The effect of both plant extracts on microbial biomass C, amount of total N and organic C, and enzymatic activity in soil was significant. After the last application, the highest microbial biomass C was determined in the lowest U. maritima concentration (U 1:100). Soils treated with the highest concentration of U. maritima (U 4:100) had always lower SMBC content than control soil. All concentrations of E. myrsinites decreased microbial biomass C by 18% to 27% compared to the control. Total nitrogen and organic carbon decreased in soils without (control) and with treated U. maritima extract from first application to last application. Phosphatase, urease, and beta-glucosidase activities were monitored in plant extract-treated soils. Except U. maritima 1:100 treatments of second and fourth applications, the other treatments of plant extracts negatively affected enzymatic activity in soil. U. maritima and E. myrsinites plant extracts exhibited different effects on soil microbial biomass and activity, probably because of their different chemical contents.

Effects of Rhodocodon madagascariensis extracts on embryo-larval development of medaka fish, Oryzias latipes.

Rhodocodon madagascariensis, also named Urginea mascarenensis, is a malagasy plant belonging to the Hyacinthaceae family. As for the other members of the endemic malagasy genus Rhodocodon, the chemical and toxicological properties of this species have not yet been studied. The present study concerns the analysis of the toxicity of R. madagascariensis to medaka embryo-larval development. The incubation of medaka fish embryos or larvae in a medium containing R. madagascariensis extract resulted in a dose dependent reduction in development of embryos leading to lethality and a drastic reduction in survival rate of exposed larvae. Survival rates were reduced up to 100% with an extract concentration of 4 mg mL(-1). The LD(50) was estimated to be 1 mg mL(-1). Anatomopathological studies did show some neuro-embryonal modifications in the encephalic region. The data presented in this paper thus extends the use of medaka embryos as a valuable model to detect and analyse the effects of plant toxins.

Bufadienolides from bulbs of Urginea lydenburgensis (Hyacinthaceae: Urgineoideae).

Bulbs of the ethnomedicinal hyacinthac Urginea lydenburgensis have yielded two bufadienolides, 16beta-acetoxy-3beta,14beta-dihydroxy-19-formyl-bufa-4,20,22-trienolide (scillicyanosidin) and 4beta,8beta,11alpha,14beta-tetrahydroxybufa-5,20,22-trienolide-12-one, 2alpha,3beta-O-4,6-dideoxy-L-glucose (lydenburgenin).

Antiangiogenic and proapoptotic activity of a novel glycoprotein from U. indica is mediated by NF-kappaB and Caspase activated DNase in ascites tumor model.

We have identified a novel glycoprotein from Urginea indica bulbs with potent in vivo antitumor activity against growth of an ascites tumor, mouse mammary carcinoma. In this paper we report characterization of a 29 kDa glycoprotein from U. indica and demonstrate the mechanism of antiangiogenic and proapoptotic activity. N-terminal sequence of the high performance liquid chromatography (HPLC) pure glycoprotein showed sequence homology to an extent of 40-50% with known angiogenesis inhibitor and apoptosis-inducing protein from C. elegans and G. gallus respectively. Our results on antiangiogenic property of the glycoprotein include inhibition of in vivo angiogenesis assays, decreased micro vessel density count and CD31 antigen staining in 29 kDa glycoprotein treated mice peritoneum. In vitro inhibition of vascular endothelial growth factor induced proliferation of human umbilical vein endothelial cells (HUVECs) by the glycoprotein further supports its antiangiogenic activity. The mechanism of antiangiogenesis involved inhibition of translocation of nuclear factor kappa B to the nucleus resulting in decreased expression of vascular endothelial growth factor gene as is demonstrated by our results on quantification of vascular endothelial growth factor levels in the glycoprotein treated tumor bearing mice. Our results on activation of Caspase-3 with concomitant translocation of caspase activated DNase to the tumor cell nuclei resulting in DNA fragmentation induced by the glycoprotein in vivo clearly demonstrated a parallel proapoptotic activity of the glycoprotein.