Formation of morpholine-acetamide derivatives as potent anti-tumor drug candidates: Pharmacological evaluation and molecular docking studies.

Heterocyclic amines and acetamide derivatives are known for their chemotherapeutic potential. Hence, in the present study, morpholine was taken as a principal product and novel morpholine derivatives were designed, formulated, characterized, and screened for the mechanism of inhibition of carbonic anhydrase and their anticancer potential. In addition, in vitro inhibition of hypoxia-inducible factor-1 (HIF-1) protein was also investigated. Results revealed that compounds 1c, 1d, and 1h possessed significant inhibitory activities against carbonic anhydrase with IC50 of 8.80, 11.13, and 8.12 µM, respectively. Interestingly, the carbonic anhydrase inhibitory activity of compound 1h was comparable with that of standard acetazolamide (IC50 7.51 µM). The compounds 1h and 1i significantly inhibited the proliferation of ovarian cancer cell line ID8 with IC50 of 9.40, and 11.2 µM, respectively while the standard cisplatin exhibited an IC50 8.50 µM. In addition, compounds 1c, 1b, 1h and 1i also exhibited significant inhibitory effects on HIF-1α. In conclusion, we report first time the biological potential of morpholine based compounds against ovarian cancer and HIF-1α that may serve as lead molecules for drug discovery.

Comparative Evaluation of Nano Inorganic Metal Oxides as Intracanal Medicaments for Cytotoxicity and Antimicrobial Activity in the Root Canal System.

Aim and objective: To evaluate and compare the cytotoxicity and antimicrobial activity of various inorganic metal oxide nanoparticles along with vehicles when used as intracanal medicaments in the root canal system. Materials and methods: The study included triplicates (n = 36 times) that were subjected to n calcium oxide (CaO), n zinc oxide (ZnO), n magnesium oxide (MgO), and metapaste as intracanal medicaments. The efficacy of novel intracanal medicaments was evaluated for biocompatibility assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reagent following antimicrobial efficacy against Enterococcus faecalis (E. faecalis) was evaluated using zone of inhibition (ZOI) and minimum inhibitory concentration (MIC). The statistical analysis Kruskal-Wallis test, student t-test, and analysis of variance (ANOVA) using Statistical Package for the Social Sciences (SPSS) software (v.20.0). Results: The order of proliferative activity of experimental groups on L929 mouse fibroblast cells using MTT assay was: metapaste > nCaO > nMgO > nZnO). After evaluation of antimicrobial efficacy, group I: nCaO showed maximum ZOI and MIC against E. faecalis, which showed high statistically significant differences between all four groups after ANOVA (p < 0.0001*). Conclusion: n calcium oxide (CaO) mixed with propylene glycol (PPG) 400 has a potential role as an intracanal medicament with minimum cytotoxic effect and maximum antimicrobial activity against endodontic pathogens. Clinical significance: Nanoparticles-based intracanal medicament can provide a promising future in reducing endodontic flareups when used as intracanal medicament. How to cite this article: Barge P, Gugawad S, Devendrappa SN, et al. Comparative Evaluation of Nano Inorganic Metal Oxides as Intracanal Medicaments for Cytotoxicity and Antimicrobial Activity in the Root Canal System. Int J Clin Pediatr Dent 2023;16(S-2):S168-S175.

Evaluation of antiangiogenic and antiproliferative potential of ethanolic extracts of Andrographis echioides using in vitro and in ovo assays.

INTRODUCTION: Andrographis echioides is a prevalently used medicinal herb in South Asian countries. Scientific researches with the extracts of A. echioides revealed its antipyretic, anti-inflammatory, antimicrobial, ulceroprotective, and hepatoprotective properties. This study was done to elucidate antiproliferative and antiangiogenic potential of ethanolic extracts of A. echioides (EEAE) by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-tetrazolium bromide (MTT) assay and chorioallantoic membrane (CAM) assay. MATERIALS AND METHODS: EEAE was prepared using Soxhlet apparatus with ethanol after being sun-dried and powdered. MCF 7 (human invasive breast ductal carcinoma) cell lines retaining attributes of differentiated mammary epithelium with both estrogen and progesterone receptors were treated with EEAE, and antiproliferative effect was seen using Mosmann method of MTT assay using 5-fluorouracil (5-FU) as a comparator. The evaluation of antiangiogenic potential of EEAE was done by comparing mean vessel density (MVD) in chick CAM after treatment with EEAE, thalidomide, and vascular endothelial growth factor (VEGF) using CAM assay, an in ovo assay. RESULTS: EEAE displayed antiproliferative activity from low to high concentrations with MTT assay. The IC50 of EEAE and 5-FU was 62.5 and 15.6 µg/ml, respectively (P < 0.05). The exhibition of its antiangiogenic activity increased proportionately with increasing concentration. VEGF increased MVD by 45.94%; thalidomide decreased it by 53.76%. There was a decrease of MVD by 5.91%, 20.46%, and 35.95% at concentrations of 25, 50, and 100 µg of EEAE, respectively. CONCLUSION: EEAE possessed significant antiangiogenic and antiproliferative activity, making them a promising substrate in the development of a novel anticancer drug and can be successfully used in the therapy of various cancers after establishment of the anticancer effects in animal models and subsequently in clinical trials.

Recombinant Human erythropoietin reduces viability of MCF-7 breast cancer cells from 3D culture without caspase activation.

Recombinant human erythropoietin (rHuEPO) is the erythropoiesis-stimulating hormone that is being used concurrently with chemotherapeutic drugs in the treatment of anemia of cancer. The effect of rHuEPO on cancer cells in 3-dimensional (3D) cultures is not known. The objective of the study was to determine the effect of rHuEPO on the viability of MCF-7 breast cancer cells from 2-dimensional (2D) and 3D cell cultures. The monolayer MCF-7 cells from 2D culture and MCF-7 cell from 3D culture generated by ultra-low adhesive microplate technique, were treated with 0, 0.1, 10, 100 or 200 IU/mL rHuEPO for 24, 48 or 72 h. The effects of rHuEPO on MCF-7 cell viability and proliferation were determined using the (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay (MTT), neutral red retention time (NRRT), trypan blue exclusion assay (TBE), DNA fragmentation, acridine orange/propidium iodide staining (AO/PI) assays. The MCF-7 cells for 3D culture were also subjected to caspase assays and cell cycle analysis using flow cytometry. rHuEPO appeared to have greater effect at lowering the viability of MCF-7 cells from 3D than 2D cultures. rHuEPO significantly (p < 0.05) decreased viability and down-regulated the caspase activities of 3D MCF-7 cells in dose- and time-dependent manner. The cell cycle analysis showed that rHuEPO caused MCF-7 cells to enter the subG0/G1 phase. Thus, the study suggests that rHuEPO has a cytostatic effect on the MCF-7 breast cancer cells from 3D culture.

In vitro cytotoxicity of cardamom oil, lemon oil, and jasmine oil on human skin, gastric, and brain cancer cell line.

OBJECTIVE: The main objective of the study was to evaluate the cytotoxicity of selected essential oils on human skin, gastric, and brain cancer cell lines using microculture tetrazolium test. MATERIALS AND METHODS: Phytochemical analysis, as well as acute oral toxicity tests, was carried out in female albino mice with cardamom oil, lemon oil, and jasmine oil according to the Organization for Economic Co-operation and Development guidelines 425. Anticancer activities of the above test drugs were performed using human cancer cell lines. The studies were carried out at Skanda Life Sciences Pvt. Ltd., Bengaluru. RESULTS: Phytochemical analysis has shown the presence of carbohydrates and flavonoids in cardamom oil. While lemon oil has shown the presence of carbohydrates, flavonoids, steroids, terpenoids, and tannins, jasmine oil has shown the presence of carbohydrates, alkaloids, flavonoids, steroids, terpenoids, and glycosides. Toxicity studies showed that cardamom oil, lemon oil, and jasmine oil were all found to be safe up to 2000 mg/kg body weight. Results have shown that lemon oil exhibited the strongest cytotoxicity toward three human cancer cell lines, namely skin cancer (A431), gastric cancer (MKN-45), and brain cancer (U-87 MG) cell lines, with higher IC50 values of 62.82 µg/ml, 220.9 µg/ml, and 440.1 µg/ml compared to standard. Jasmine oil exhibited the strongest cytotoxicity toward skin cancer and brain cancer cell lines, whereas cardamom oil has shown stronger cytotoxicity only toward skin cancer cell line but did not show any level of inhibition of growth of brain and gastric cancer cells. CONCLUSION: Our study reveals that lemon oil, jasmine oil, and cardamom oil possess potent antitumor activity compared to standard. At different concentrations, lemon oil has shown statistically significant (***P < 0.0001) anticancer activity toward all the three human cancer cell lines. While jasmine oil has shown statistically significant (***P < 0.0001) anticancer activity toward skin and brain cancer cell line, cardamom oil has also shown statistically significant (***P < 0.0001) anticancer activity but only toward skin cancer cell line.

Cytotoxicity of oregano essential oil and calcium hydroxide on L929 fibroblast cell: A molecular level study.

Introduction: The purpose of antimicrobial agents is to eliminate the microorganisms without causing toxicity to host cells. This study aimed to assess the cytotoxic effect of oregano essential oil on L929 fibroblast cells. Materials and Methods: L929 fibroblast cells were exposed to four different concentrations of oregano essential oil (25-200 µg/ml) and calcium hydroxide (1 mg/ml). Dose-response curve was evaluated using 3-(4,5 dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay. The toxicity of L929 fibroblast cells was determined by lactate dehydrogenase activity (LDH). The results were analyzed using one-way analysis of variance and turkey post hoc test. P <0.05 was considered statistically significant. Results: Oregano essential oil showed a higher percentage of cell viability than the calcium hydroxide group. At 50 µg/ml, fibroblast cells showed arbitrarily 80% of cell viability compared to calcium hydroxide. There was a statistically significant difference with P < 0.05 on evaluating the effect of oregano essential oil on cytotoxicity measurement by LDH release of L929 fibroblast cells. Conclusion: Within the limitation of the study, Oregano essential oil at 50 µg/ml reported to show reduced cytotoxicity compared to calcium hydroxide at 1 mg/ml. Therefore, perhaps after evaluating other properties, it might be considered an intracanal medicament.

In Vitro Antimalarial and Toxicological Activities of Quercus infectoria (Olivier) Gall Extracts.

BACKGROUND: The spread of Plasmodium falciparum resistance in common antimalarial drugs, including artemisinin-based combination therapies, has necessitated the discovery of new drugs with novel mechanisms of action. In the present study, the in vitro antimalarial and toxicological activities of acetone, methanol, ethanol and aqueous extracts of Quercus infectoria (Q. infectoria) galls were investigated. METHODS: The extracts were assessed for the antimalarial potential using a malarial SYBR Green I fluorescence-based (MSF) assay, while the toxicity was screened by using brine shrimp lethality test (BSLT), haemolytic assay, and cytotoxicity assay against normal embryo fibroblast cell line (NIH/3T3) and normal kidney epithelial cell line (Vero). RESULTS: The acetone extract showed the highest antimalarial activity (50% inhibitory concentration, IC50 = 5.85 ± 1.64 µg/mL), followed by the methanol extract (IC50 = 10.31 ± 1.90 µg/mL). Meanwhile, the ethanol and aqueous extracts displayed low antimalarial activity with IC50 values of 20.00 ± 1.57 and 30.95 µg/mL ± 1.27 µg/mL, respectively. The significant antimalarial activity was demonstrated in all extracts and artemisinin (P < 0.05). All extracts were non-toxic to brine shrimps (50% lethality concentration, LC50 > 1000 ppm). Furthermore, no occurrence of haemolysis (< 5%) was observed in normal erythrocytes when treated with all extracts compared to Triton X-100 that caused 100% haemolysis (P < 0.05). The acetone and methanol extracts were non-toxic to the normal cell lines and statistically significant to artemisinin (P < 0.05). CONCLUSION: Taken together with satisfactory selectivity index (SI) values, the acetone and methanol extracts of Q. infectoria galls could serve as an alternative, promising and safe antimalarial agents.

Effect of Ocimum sanctum extract on leukemic cell lines: A preliminary in-vitro study.

BACKGROUND: Surge of cancer incidence, effects of chemotherapeutic agents and their cost and reduced survival and responsiveness to treatment have led to shift of attention of researchers toward herbal remedies to look for newer dimension in cancer therapy. Ocimum sanctum, Holy Basil or Tulsi, holiest herb well used in the Indian household, has drawn much attention toward its various health benefits, especially anti-cancer property. The present study was carried out to evaluate the cytotoxic effect of O. sanctum on leukemic cell lines K562. MATERIALS AND METHODS: Dry and aqueous extracts of two types of Tulsi leaves (Rama Tulsi and Krishna Tulsi) were evaluated for a dose-dependent cytotoxicity and anti-proliferative against K562 cell lines, leukoerythroid progenitor leukemic cell lines by 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Half-maximal inhibitory concentration was evaluated for each of the extracts. RESULTS: Both dry and aqueous extracts of both types of Tulsi leaves demonstrated a significant amount of cytotoxicity against the studied cell lines. CONCLUSION: This being preliminary study, we propose the initial finding of cytotoxic abilities of the herb against the leukemic cell lines and recommend a more detailed evaluation of the herb and its components.

Evaluation of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide method for assessing biofilm formation in vitro by Trichosporon spp.

BACKGROUND: Invasive infections due to Trichosporon spp. have increased recently and are frequently associated with indwelling medical devices. Such infections which are associated with biofilm formation do not respond to the routinely used antifungal agents and are often persistent, associated with high mortality rate. Various methods have been described by researchers to evaluate and quantify the biofilm formation. AIM: This study was conducted to compare two methods of biofilm production by Trichosporon sp, i.e., test tube method with crystal violet (CV) staining and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. MATERIALS AND METHODS: Seventy-two clinical isolates of Trichosporon spp. collected from various sources were considered for the study. The identity of all the isolates was genotypically confirmed by Trichosporon-specific polymerase chain reaction (PCR). The isolates were further speciated phenotypically using biochemical profile and growth characteristics which identified the isolates as Trichosporon asahii (64/72), Trichosporon asteroides (5/72), Trichosporon cutaneum (2/72), and Trichosporon mucoides (1/72). Biofilm production was then evaluated and compared by test tube-CV method and MTT assay. RESULTS: All the Trichosporon isolates produced biofilm by MTT assay, whereas only 42 (53.6%) of the isolates were detected to be biofilm producers by CV method. Furthermore, MTT assay could differentiate better between weak and moderate biofilm producers as compared to CV method. CONCLUSION: Hence, MTT assay is a reliable method for quantification of biofilm produced by Trichosporon spp. using 96-well microtiter plate.

Comparative Evaluation of Cytotoxicity of Root Canal Sealers on Cultured Human Periodontal Fibroblasts: In vitro Study.

AIM: To evaluate and compare the cytotoxic effects of different types of root canal. MATERIALS AND METHODS: The sealers were eluted with culture medium for 1 hour, 7 days, and 14 days. Cell viability was estimated by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay and trypan blue exclusion method on human periodontal ligament (PDL) fibroblast cells. Sealers used are mineral trioxide aggregate (MTA)-based sealer (MTA Fillapex, Angelus), calcium hydroxide-based sealer (Apexit Plus, Ivoclar Vivadent), resin-based sealer (AH Plus, Dentsply), and zinc oxide eugenol-based sealer (Tubli Seal, SybronEndo). RESULTS: The order of cytotoxicity through MTT assay, at the end of the second week, was observed as MTA Fillapex> Tubli Seal> Apexit Plus > AH Plus. The percentage cell viability obtained after trypan blue exclusion method decreased in the order of Apexit Plus> Tubli Seal> AH Plus> MTA Fillapex, which was similar to the reported cytotoxicity from the MTT assay after 1 hour. CONCLUSION: Each type of sealer showed moderate-to-severe cytotoxic response when compared with the control. The MTA Fillapex was found to be the most cytotoxic sealer. Use of resin-based material as a root canal sealer may result in a more favorable response to PDL fibroblasts. CLINICAL SIGNIFICANCE: Having knowledge of the cytotoxicity of various sealers will help in increasing patient's comfort.

Antihyperglycemic Activity of Caralluma fimbriata: An In vitro Approach.

BACKGROUND: An increase in prevalence of diabetes mellitus necessitates the need to develop new drugs for its effective management. Plants and their bioactive compounds are found to be an alternative therapeutic approach. Caralluma fimbriata, used in this study, is well known for its various biological effects. OBJECTIVE: The present study was designed to investigate the antihyperglycemic effect of the ethanolic leaf extract of C. fimbriata. MATERIALS AND METHODS: Different concentrations (1-1000 mg/mL) of the ethanolic leaf extract of C. fimbriata were subjected to alpha-amylase and alpha-glucosidase inhibitory assay with acarbose as control. Cytotoxicity was assessed by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Glucose uptake assay was performed on L6 myotubes using the extract in 1 µg-100 µg/mL, using metformin and insulin as control. RESULTS: The C. fimbriata extract showed potent inhibitory activity on enzymes of glucose metabolism in a dose-dependent manner. The maximum alpha-amylase inhibitory effect was 77.37% ± 3.23% at 1000 µg/mL with an IC50 value of 41.75 µg/mL and alpha-glucosidase inhibitory effect was 83.05% ± 1.69% at 1000 µg/mL with an IC50 value of 66.71 µg/mL. The maximum glucose uptake was found to be 66.32% ± 0.29% for the Caralluma extract at 100 µg/mL and that of metformin (10 µg/mL) was 74.44% ± 1.72% and insulin (10 mM) 85.55% ± 1.14%. The extract was found to be safe as the IC50 of extract and metformin was found to be ≥1000 µg/mL and ≥1000 µM, respectively, in the cell line tested. CONCLUSION: The study concludes that C. fimbriata has promising antihyperglycemic activity. SUMMARY: Caralluma fimbriata extract exhibited effective dose dependent inhibitory activity against alpha-amylase and alpha- glucosidaseEnhanced glucose uptake from L6 myotubes was appreciated in the presence of the extract, comparable to Insulin and metforminCaralluma fimbriata has potent antihyperglycemic properties. Abbreviations used: GLUT: Glucose transporter; MTT: 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide.

A comparative study of the in vitro activity of iodopropynyl butylcarbamate and amphotericin B against Prototheca spp. isolates from European dairy herds.

The objective of this study was to assess the in vitro effect of iodopropynyl butylcarbamate (IPBC) and amphotericin B (AMB) on Prototheca zopfii genotype 2 and Prototheca blaschkeae isolates recovered from dairy herds of Belgium, France, Italy, Germany, and Poland. The combination of IPBC with AMB on Prototheca isolates and toxicity of IPBC to the bovine mammary epithelial cells were also evaluated. The in vitro activity of IPBC and AMB against 96 isolates of P. zopfii genotype 2 and 42 isolates of P. blaschkeae was performed. Minimum inhibitory concentrations (MIC) and minimum algicidal concentrations (MAC) of IPBC and AMB were determined. To determine any synergistic, additive, or antagonistic effect of the combination of IPBC and AMB, 2-dimensional checkerboard combination tests were also performed to calculate fractional inhibitory concentrations. Cytotoxicity analysis of IPBC to the bovine mammary epithelial cell line was performed using a 3-(4,5-dimethyl-2-thiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The MIC for 50 and 90% of isolates (MIC50 and MIC90, respectively) for IPBC were 4 and 8 mg/L versus 0.5 and 1 mg/L for AMB, respectively. The MIC profiles differed between P. zopfii genotype 2 and P. blaschkeae, with the latter species being more susceptible to both compounds. The MIC50 and MIC90 of IPBC were 4 and 8 mg/L for P. zopfii genotype 2 and 1 and 2 mg/L for P. blaschkeae, respectively. The MIC50 and MIC90 of AMB were both 1 mg/L for P. zopfii genotype 2 and 0.25 and 1 mg/L for P. blaschkeae, respectively. Both IPBC and AMB exhibited the ability to kill Prototheca spp. The MAC for 90% of isolates of IPBC was twice the MIC90, whereas an 8-fold increase of the MIC90 was algicidal in the case of AMB. Overall, the combined use of IPBC and AMB exhibited an increased algicidal effect, albeit the fractional inhibitory concentration index showed synergistic activity only against 3 P. zopfii genotype 2 isolates. For all the remaining isolates (87.5%), this combination produced only an additive effect. The MTT assay results showed both IPBC and AMB, at the concentrations employed in the study, to be nontoxic to the epithelial mammary gland cells (cell viability >90%). Notably, only IPBC at the highest concentration (i.e., 8 mg/L) exerted a slight cytotoxic effect on the cell line tested (mean cell viability: 88.54 ± 3.88 and 90.66 ± 3.0, after 2 and 4 h of MTT treatment, respectively). The anti-Prototheca activity of IPBC was here demonstrated for the first time. In addition, the combined use of IPBC with AMB enhanced each other's effect, creating an additive rather than synergistic interaction. Both agents, used at concentrations corresponding to MIC values against Prototheca spp., showed no toxic effect for the mammary epithelial cells. In conclusion, IPBC, used either alone or in combination with AMB, can be considered a promising option in the treatment armamentarium for protothecal mastitis in dairy cows.

Assessment of cytotoxicity of Portulaca oleracea Linn. against human colon adenocarcinoma and vero cell line.

BACKGROUND: Portulaca oleracea Linn. (Portulacaceae) is commonly known as purslane in English. In traditional system it is used to cure diarrhea, dysentery, leprosy, ulcers, asthma, and piles, reduce small tumors and inflammations. AIM: To assess cytotoxic potential of chloroform extract of P. oleracea whole plant against human colon adenocarcinoma (HCT-15) and normal (Vero) cell line. MATERIALS AND METHODS: Characterization of chloroform extract of P. oleracea by Fourier transform infrared (FTIR) spectroscopy was performed. Cytotoxicity (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay was used for assessment of cytotoxic potential of chloroform extract of P. oleracea. The concentrations of 1000-0.05 µg/ml were used in the experiment. Doxorubicin was considered as standard reference drug. RESULTS: FTIR spectrum showed the peak at 1019.52 and 1396.21 center. The 50% cell growth inhibition (IC50) of chloroform extract of P. oleracea and doxorubicin was 1132.02 µg/ml and 460.13 µg/ml against human colon adenocarcinoma and 767.60 µg/ml and 2392.71 µg/ml against Vero cell line, respectively. CONCLUSION: Chloroform extract of P. oleracea whole plant was less efficient or does not have cytotoxic activity against human colon adenocarcinoma cell line. It was not safe to normal Vero cell line. But, there is a need to isolate, identify, and confirm the phytoconstituents present in extract by sophisticated analytical techniques.

Cytotoxicity of different extracts of arial parts of Ziziphus spina-christi on Hela and MDA-MB-468 tumor cells.

BACKGROUND: It has been shown that plants from the family Rhamnaceae possess anticancer activity. In this study, we sought to determine if Ziziphus spina-christi, a species from this family, has cytotoxic effect on cancer cell lines. MATERIALS AND METHODS: Using maceration method, different extracts of leaves of Z. spina-christi were prepared. Hexane, chloroform, chloroform-methanol (9:1), methanol-water (7:1) methanol, butanol and water were used for extraction, after preliminary phytochemical analyses were done. The cytotoxic activity of the extracts against Hela and MDA-MB-468 tumor cells was evaluated by MTT assay. Briefly, cells were seeded in microplates and different concentrations of extracts were added. After incubation of cells for 72 h, their viability was evaluated by addition of tetrazolium salt solution. After 3 h medium was aspirated, dimethyl sulfoxide was added and absorbance was determined at 540 nm with an ELISA plate reader. Extracts were considered cytotoxic when more than 50% reduction on cell survival was observed. RESULTS: Hexane, chloroform, chloroform-methanol, butanol, methanol-water and aqueous extracts of Z. spina-christi significantly and concentration-dependently reduced viability of Hela and MAD-MB-468 cells. In the both cell lines, chloroform-methanol extract of Z. spina-christi was more potent than the other extracts. RESULTS: From the finding of this study it can be concluded that Z. spina-christi is a good candidate for further study for new cytotoxic agents.