Standard Vincristine Sulphate Research Articles

Anthelmintic and Cytotoxic Activities of Two Medicinal Plants: <i>Polygonum viscosum</i> and <i>Aphanamixis polystachya</i> Growing in Bangladesh

The present study was designed to investigate in vitro anthelmintic and cytotoxic activities of crude methanolic extract of two plants(Polygonum viscosum and Aphanamixis polystachya) grown in Bangladesh. Evaluation of cytotoxic activity was done using the brine shrimp lethality bioassay. The crude methanolic extract of Polygonum viscosum showed significant cytotoxic potential (LC50 value of 6.34 ?g/ml) among all the fractions comparing with that of standard vincristine sulphate (0.825 ?g/ml). Besides, the LC50 values of crude methanolic extract, pet ether and chloroform extracts of Aphanamixis polystachya showed good cytotoxic activities 11, 10.36, and 16.45 µg/ml, respectively. The other study was undertaken to evaluate anthelmintic activity (using Pheretima posthuma model) where piperazine was used as reference standard. The crude methanolic extract of Polygonum viscosum leaves produced a significant anthelmintic activity in dose dependent manner and the activity of crude extract was comparable with that of standard drugs. Besides, the Aphanamixis polystachya extract revealed moderate anthelmintic activity. Here, the anova testing was done with the P < 0.05. Further studies are suggested to determine the active compounds responsible for the anthelmintic and cytotoxic activities of these two plant extracts. Keywords: Anthelmintic; Cytotoxic; Medicinal plant; Aphanamixis polystachya; Polygonum viscosu. © 2014 JSR Publications. ISSN: 2070-0237 (Print); 2070-0245 (Online). All rights reserved. doi: http://dx.doi.org/10.3329/jsr.v6i2.17299 J. Sci. Res. 6 (2), 339-345 (2014)

Preliminary phytochemical, cytotoxic, thrombolytic and antioxidant activities of the methanol extract of Murraya exotica Linn. leaves

The methanol extract of leaves of traditionally used medicinal plant Murraya exotica Linn. (Family: Rutaceae) was evaluated for possible cytotoxic, thrombolytic and antioxidant activities in this study. The extract was tested for phytochemical group test by standard method using chromogenic reagents, in vivo brine shrimp lethality assay using Artemia salina, thrombolytic effect by using the standard streptokinase and antioxidant activity by using 2,2-diphenyl-1-picryl-hydrazyl (DPPH). The study revealed the presence of reducing sugars, tannins, saponins and alkaloids. The extract showed statistically significant (p < 0.01) potent cytotoxic effect in brine shrimp lethality bioassay where the value of LC50 and LC90 were 1.27 μg/ml and 5.09 μg/ml respectively compared with the standard vincristine sulphate with the value of LC50 and LC90 0.09 μg/ml and 4.83 μg/ml respectively after 24 h. The study gave a significant indication to the use of the plant extract as a potential source for cytotoxic compounds. The extract showed mild thrombolytic effect of 15.78 % thrombolytic activity whereas the standard streptokinase showed 76.50 ± 0.82 %. In the antioxidant activity study, the extract showed free radical scavenging activity where IC50 = 1.25 μg/ml and IC90 = 4.4 μg/ml, compared with the standard ascorbic acid showing IC50 = 0.01 μg/ml and IC90 = 3.58 μg/ml. Based on the findings of phytochemical, toxicological and anti-oxidative activity, it is evident that the plant may contain some novel compounds that possess potent anti-mutagenic and anti-oxidative activities. The obtained results support the use of this plant in traditional medicine.

PHARMACOLOGICAL INVESTIGATION OF THE CHLOROFORM EXTRACTS OF ALSTONIA SCHOLARIS (L.) R.BR

Chloroform extracts of Alstonia scholaris leaves (CLAS ) and stem barks ( CSAS ) have been evaluated for their analgesic, antimicrobial, antioxidant and cytotoxic activity. Acetic acid induced writhing method was used for evaluating analgesic activity . Both the extracts had reduced pain in dose dependent manner, at all the test ed doses (200 and 400 mg/kg body weight) . Maximum writhing inhibition (75.33 %) was observed at 400 mg/kg dose of CSAS while for CLAS, 400 mg/kg dose exhibited 70.13 % inhibition . The inhibitory effect of indomethacin (45.92 %) was lower than that of the highest dose of CLAS and CSAS. Antimicrobial activity of the extract s was evaluated against various Gram - positive, Gram - negative bacteria and fungi using disk diffusion technique. The average zone of inhibition exhibited by extract was found 10 - 1 4 mm and Kanamycin (30 μg/disc) was used as standard. Antioxidant potentiality of the extracts was investigated on DPPH scavenging activity and t he IC 50 value was found 47.72 µg/ml, 62.03 µg/ml and 45.77 µg/ml for CSAS, CLAS and standard ascorbic acid , respectively. Cytotoxic st udy was done by brine shrimp lethality bioassay and compared with LC 50 ( 8.90 μg/ml. ) values of standard vincristin sulphate as a positive control. The cytotoxicity exhibited CLAS and CSAS were promising with LC 50 value of 10.21 μg/ml and 9.12 μg/ml, respec tively. The se results suggest into the plant extracts could be used as a potential therapeutics in many pathological conditions.

In vivo sedative and cytotoxic activities of methanol extract of leaves of Crataeva nurvala Buch-Ham.

The present study was designed to investigate the sedative and cytotoxic activities of a crude methanol extract of leaves of Crataeva nurvala Buch-Ham. Sedative activity was evaluated by using hole cross, open field and Elevated-Plus Maze (EPM) tests at 400 mg kg(-1) body weight. The crude extract decreased the locomotor activity of mice in hole cross, open field and EPM tests. The cytotoxic activity of this extract was determined by brine shrimp lethality bioassay where the LC50 value was found to be 55.46 microg mL(-1) as compared to that of 0.451 microg mL(-1) exhibited by standard vincristine sulphate. The result shows that the crude extract of the leaves of C. nurvala have significant (* p < 0.05) sedative and cytotoxic activities.

Evaluation of Antioxidant, Cytotoxic, Antimicrobial, Membrane Stabilizing and Thrombolytic Activities of Polianthes tuberose Linn

Aims: In the present study, the crude methanol extract of tuber of Polianthes tuberosa Linn along with its all Kupchan fractions were investigated for antioxidant, cytotoxic, antimicrobial, membrane stabilizing and thrombolytic activities. Place and Duration of Study: The study was carried out for one year in 2012 in the Department of Pharmacy, Manarat International University (MIU), Dhaka-1216, Bangladesh. Methodology: The antioxidant activity was evaluated by using free radical scavenging (DPPH) assay. Here, butylated hydroxytoluene (BHT) was used as standard antioxidant. The total phenolic content was also determined and expressed in gallic acid equivalent. Cytotoxicity and antimicrobial activity of the plant fractions were determined by brine shrimp lethality bioassay as well as by the disc diffusion method, respectively. The membrane stabilizing activity was assessed by hypotonic solution and heat-induced methods and was compared with standard acetyl salicylic acid (ASA). Results: In the free radical scavenging assay, the crude methanol extraxct showed significant free radical scavenging activity with IC50 value 71.23 μg/ml. The highest phenolic content was found in crude methanol extract (113.49 mg of GAE/gm of extractives). In the brine shrimp lethality bioassay, both the crude methanol extract and its carbon tetrachloride soluble fraction demonstrated strong cytotoxic activity with LC50 value of 3.56 and 9.31 μg/ml, respectively compared to that of 0.451 μg/ml exhibited by standard vincristine sulfate (VS). In the disc diffusion antibacterial assay, all the plant samples showed mild to moderate activity (zone of inhibition = 9.0-15.0 mm) against test pathogens. In membrane stabilizing activity test, the plant samples at 2.0 mg/ml inhibited the isotonic solution-induced hemolysis of RBC by 65.23% and heat-induced hemolysis of RBC by 35.61%. During assay for thrombolytic activity, the crude methanol extract revealed 52.6% lysis of clot while standard streptokinase (SK) used as positive control, demonstrated 66.8% lysis of clot. Conclusion: The plant possesses significant bioactivities which rationalize its use as folk medicine.

Ex-Vivo Cardioprotective and Cytotoxic Screening of Fruits of Parmentiera cereifera Seem

In this present investigation, the fruits extract of Parmentiera cereifera were subjected to a comparative evaluation of the cardio-protective activities and Cytotoxic to brine shrimps (Artemia salina) as compared with standard streptokinase (SK) and Vincristine sulphate (VS). The crude methanolic (ME) extract, Hexane (HXSF) soluble fraction, carbon tetrachloride (CTCSF) soluble fraction, the Aqueous (AQSF) and chloroform (CSF) soluble fraction fruits of P. cereifera were subjected to these studies. In thrombolytic study, the fruits extract crude methanolic (ME) extract and Hexane (HXSF) soluble fraction were demonstrated mild to moderate (2.5 to 21.50%) thrombolytic activity in human blood specimen compared with standard streptokinase (SK) 66.77%. The fruits extracts were demonstrated (1.78 to 79.90) significant Cytotoxic to A. salina with LC50 values ranging from 6.07 to 7.83 µg/mL compared with standard Vincristine sulphate (VS, LC50 value 0.45 µg/mL).

Cytotoxicity potentials of eleven Bangladeshi medicinal plants.

Various forms of cancer are rising all over the world, requiring newer therapy. The quest of anticancer drugs both from natural and synthetic sources is the demand of time. In this study, fourteen extracts of different parts of eleven Bangladeshi medicinal plants which have been traditionally used for the treatment of different types of carcinoma, tumor, leprosy, and diseases associated with cancer were evaluated for their cytotoxicity for the first time. Extraction was conceded using methanol. Phytochemical groups like reducing sugars, tannins, saponins, steroids, gums, flavonoids, and alkaloids were tested using standard chromogenic reagents. Plants were evaluated for cytotoxicity by brine shrimp lethality bioassay using Artemia salina comparing with standard anticancer drug vincristine sulphate. All the extracts showed potent to moderate cytotoxicity ranging from LC50 2 to 115 µg/mL. The highest toxicity was shown by Hygrophila spinosa seeds (LC50 = 2.93 µg/mL) and the lowest by Litsea glutinosa leaves (LC50 = 114.71 µg/mL) in comparison with standard vincristine sulphate (LC50 = 2.04 µg/mL). Among the plants, the plants traditionally used in different cancer and microbial treatments showed highest cytotoxicity. The results support their ethnomedicinal uses and require advanced investigation to elucidate responsible compounds as well as their mode of action.

Evaluation of In vitro Anthelmintic Activity, Total Phenolic Content and Cytotoxic Activity of Crinum latifolium L. (Family: Amaryllidaceae).

Crinum latifolium is a widely used plant in Asian folk and traditional medicine.In the present study, we have tried to find out the anthelmintic activity, total phenolic contents and cytotoxicity of the methanolic extract of the target plant. Anthelmintic activity was assessed applying five different concentrations of the plant extract and recording the time of paralysis and death. Total phenolic contents were determined using Folin-Ciocaltu method, using Gallic acid as standard; while brine shrimp lethality test (BSLT) method was used to evaluate the cytotoxicity of the plant extract, where vincristine sulphate and DMSO was used as positive and negative control respectively. The lowest time for paralysis and death of worms, for test sample at highest concentration (50mg/ml), were found 24±0.45 and 46.4±0.60 min respectively, which gradually increased with the decrease of concentration. On the other hand, albendazole, which was used as standard, caused paralysis and death of worms at 56.2±0.20 min and 77.4±0.24 min respectively; whereas no mortality of the worms was observed, when distilled water was used as control. The crude methanolic extract exhibited lower amount of total phenolic content (17.50±2.64 mg/ml). In case of cytotoxicity measurement, the crude methanolic extract showed positive result (with LC50 15.652 µg/ml) compared to standard Vincristine sulphate (0.839 µg/ml); which indicated that the leaves of Crinum latifolium possess mild cytotoxic principles. Therefore, further studies are suggested to evaluate the possible mechanism of action and the active compounds responsible for the biological activities of the plant extract.

Screening and evaluation of antioxidant, antimicrobial, cytotoxic, thrombolytic and membrane stabilizing properties of the methanolic extract and solvent-solvent partitioning effect of Vitex negundo Bark

ObjectiveTo investigate different biological activities of the methanolic extract and solvent-solvent partitioning of Vitex negundo (V. negundo) bark. MethodsIn-vitro anti-oxidant study was determined using total phenolic compound analysis and DPPH radical scavenging assay. In vitro antimicrobial study was measured by observing zone of inhibition. The cytotoxic activity was studied using brine shrimp lethality bioassay and thrombolytic activity by clot disruption method. ResultsIn the evaluation of anti-oxidant activity, the amount of total phenolic content differed in different extractives as from 12.94 mg of GAE/g of extractives to 54.19 mg of GAE/g of extractives of V. negundo. Among all extractives, the highest phenolic content was found in n-hexane fraction (N-HXN) (54.19 mg of GAE/g of extractives). Significant amount of phenolic compounds were present in methanol extract (ME) (35.19 mg of GAE/g of extractives) and aqueous soluble fraction (AQSF) (12.94 mg of GAE/g of extractives). The free radical scavenging activity of ME and its different partitions were assessed using DPPH free radicals assay. The highest free radical scavenging activity was found in AQSF (IC50=7.78 μg/mL). The antimicrobial screening of the bark of V. negundo exhibited mild to moderate activity in test microorganisms. The chloroform soluble fraction demonstrated the highest inhibition against microbial growth having zone of inhibition as 8-12 mm. In the brine shrimp lethality bioassay, The LC50 values of n-hexane and chloroform soluble fractions were found 3.713 6 μg/mL, and 0.910 μg/mL respectively while the LC50 values of standard Vincristine sulphate was 0.408 μg/mL. The methanol extract and their different organic soluble of V. negundo bark inhibited hypotonic solution-induced haemolysis of erythrocytes in In-vitro membrane stabilizing activity test. The AQSF and N-HXN showed inhibition of haemolysis as 55.05% and 29.89% that were lower than 71.9% of aspirin (0.10 mg/mL). Methanol extract of V. negundo and all of its different partitions exhibited moderate thrombolytic activity of 36.95%-22.98%. ConclusionsThese experiments were able to show the several biological activity of methanolic extract and its soluble fractions of V. negundo at a time.

Abstract 3356: Comparative plasma and cerebrospinal (CSF) pharmacokinetics (PK) of vincristine sulfate standard preparation (VCR) and vincristine sulfate liposomes injection (VSLI) in a non-human primate (NHP) model.

Abstract Background: Vincristine has been used as standard therapy for adult and pediatric malignancies for 50 years. Cumulative neurotoxicity often requires capping the maximum single and cumulative VCR dose. VSLI is a novel preparation of VCR encapsulated in sphingomyelin/cholesterol liposomes. Following clinical trials that demonstrated safety, tolerability and efficacy in adults with relapsed/refractory acute lymphoblastic leukemia, VSLI has been granted FDA accelerated approval at a dose of 2.25 mg/m2given once weekly for that population. A pediatric Phase I trial is ongoing. We evaluated plasma and CSF PK of intravenously (IV) administered VCR and total VSLI (liposomal encapsulated and non-encapsulated) in our NHP model. Methods: 0.1 mg/kg (1.2 mg/m2 human-equivalent dose) VCR over 1 minute or VSLI over 15 minutes was administered on separate occasions to each of 3 adult rhesus monkeys (Macaca mulatta). Serial paired plasma and CSF samples were collected for up to 48 hrs. Drug concentrations were quantified using HPLC/tandem mass spectrometry assays. For plasma samples, VCR standard curves were linear over a range of .0025-10.0 μg/ml (lower limit of quantification, .0025 μg/ml). Lower limit of detection for VCR in NHP CSF was .001 μg/ml. PK parameters were estimated using non-compartmental methods. Results: Plasma PK parameters (median and range) for VCR and VSLI Cmax (μg/ml) AUC0-infinity (μg*hr/ml) Cl (ml/min) Terminal T1/2 (hours) VCR 1.07 (0.99–2.1) 0.614 (0.336–0.676) 33.8 (29.6–54) 24.3 (14.2–54.4) VSLI 4.8 (3.7–7.3) 195 (163–312) 0.089 (0.073–0.113) 17.9 (13.9–21.5) Cmax = maximal concentration, AUC = area under the concentration x time curve, Cl = clearance, T1/2 = half life VCR demonstrated a multi-exponential and VSLI a near-monoexponential plasma concentration versus time curve. The ratios ClVCR: ClVSLI for the individual NHP were 300, 463 and 477. Vincristine was not detected in any CSF sample. Conclusions: The NHP model allowed for a direct comparison of VCR and VSLI PK. VCR Cl exceeds that of VSLI by approximately 400-fold. No penetration into the CSF was observed for either preparation. Citation Format: Diane E. Cole, Cynthia M. Lester-McCully, Nirali N. Shah, Katherine E. Warren, Alan S. Wayne, Brigitte C. Widemann. Comparative plasma and cerebrospinal (CSF) pharmacokinetics (PK) of vincristine sulfate standard preparation (VCR) and vincristine sulfate liposomes injection (VSLI) in a non-human primate (NHP) model. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3356. doi:10.1158/1538-7445.AM2013-3356

In vitro antioxidant, cytotoxic, thrombolytic activities and phytochemical evaluation of methanol extract of the A. philippense L. leaves

ObjectiveTo study the leaves of Adiantum philippense L. for their antioxidant, cytotoxicity and thrombolytic activities and to perform phytochemical evaluation. MethodsIn-vitro antioxidant activity of extract was studied using DPPH radical scavenging, reducing power, total phenol and total flavonoid content determination assays. The cytotoxic activity was determined using brine shrimp lethality bioassay, thrombolytic activity by clot disruption and phytochemical potential by qualitative analysis. ResultsThe antioxidant activity of the extracts was found promising. The reducing power of this crude extract increase with the increase of concentration; IC50 values of DPPH scavenging activity was (140.00±0.86) μg/mL as compared to ascorbic acid [IC50 (130.00±0.76) μg/mL]; Total phenol and total flavonoids content were (148.26±0.24) mg/mL and (163.06±0.56) mg/mL respectively. In cytotoxicity assay the LC50 values of the sample was (106.41±0.78) μg/mL where as for standard vincristin sulphate was (08.50±0.24) μg/mL as a positive control and the extract shows (12.86±1.02)% clot lytic whereas standard streptokinase shows (30.86±0.44% clot lytic activity in thrombolytic assay. The phytochemical evaluation indicates the presence of chemical constituents including carbohydrates, alkaloids, saponins, glycosides, flavonoids. ConclusionsThis study shows that the methanol extract of leaves of Adiantum philippense L. has bioactivity but further compound isolation is necessary to confirm the activities of individual compounds.

Vincristine Sulfate Liposome Injection (VSLI, Marqibo®) Facilitates Increased Delivery of Vincristine Sulfate to Target Cancer Tissues.

AbstractAbstract 2457 Background:Standard vincristine sulfate (VCR) plasma pharmacokinetics (PK) is described by a bi-exponential profile with a very short half-life followed by a longer elimination half-life; the volume of distribution is large, suggesting wide and diffuse distribution and perhaps tissue binding. These PK characteristics may limit optimal therapeutic activity of VCR by limiting Cmaxand drug exposure in target tissues involved with cancer. VinCRIStine sulfate LIPOSOME injection (VSLI, Marqibo) is a sphingomyelin- and cholesterol-based nanoparticle formulation of VCR that was designed to be different from and overcome the dosing and pharmacokinetic limitations of standard VCR. The objectives for development of VSLI were to: 1) increase the plasma circulation time; 2) increase tumor tissue delivery by preferential extravasation from fenestrated (“leaky”) vasculature; 3) accumulation in tumor tissues; and 4) slow release of VCR in tumor tissues instead of the systemic circulation. VSLI recently received accelerated FDA approval for use in adults with advanced, relapsed and refractory Philadelphia chromosome negative ALL and is in development for pediatric ALL and aggressive NHL. Methods:The PK and tissue distribution of VCR was evaluated in Sprague-Dawley rats following a single IV bolus dose (2.0 mg/m2) of VSLI or VCR, utilizing 3H-VCR as a marker. The tissue levels of total VCR were measured for up to 72 hours post-dose. PK analysis was performed using noncomparmental methods with the WinNonlin software package. Results:The PK profile and calculated parameters of VSLI in rats showed a substantially lower total vincristine clearance (CL) and volume of distribution (Vdz) and correspondingly greater area under the plasma concentration versus time curve (AUC) compared to standard VCR. VSLI has a long circulation time and remains in the plasma instead of being widely distributed in tissues. For most tissues from VSLI treated animals, tissue to plasma concentration ratios increased over time and peaked at 72 h after VSLI injection, indicating progressive accumulation of radiolabeled drug from plasma into the tissues. The rank order of tissues based on Cmax demonstrated that total VCR concentrations in MPS tissues (i.e., spleen, liver, lymph nodes and bone marrow) and in ovaries were substantially higher than in other organs or tissues. The lowest radioactivity levels were observed in brain, spinal cord, nerves and muscle. Higher exposures, as measured by AUCinf, of VCR were observed in plasma (83 fold), spleen (12 fold), lymph nodes (10 fold), liver (4 fold) and bone marrow (2 fold) following a radiolabeled dose of VSLI compared to VCR.AUCinf, ng eq*hr/gSpleenLiverLymph nodeBone marrowVSLI381,15428,20266,33340,936VCR30,7656,4966,42017,767 Conclusion:The long circulation time and small, 100 nm, mean nanoparticle size of VSLI facilitate extravasation from fenestrated vasculature and accumulation in tissues involved in hematologic malignancies such as lymph nodes, bone marrow and spleen. Slow release of VCR in target tissues following administration of VSLI results in higher and prolonged tissue drug levels providing superior drug delivery to tissues than the same doses of standard formulation of VCR. Disclosures:Silverman:Talon Therapeutics: Employment. Deitcher:Talon Therapeutics: Employment, Equity Ownership.

Preliminary In Vitro Biological and Phytochemical Screenings of Parmentiera cereifera Seem

In this present study, the leaf extracts of Parmentiera cereifera were subjected to a comparative evaluation of the antioxidant capacities, phenol and polyphenols content, membrane stabilization, thrombolytic, antimicrobial activities and cytotoxicity to brine shrimps (Artemia salina). When the DPPH (1,1- Diphenyl-2-picryl hydrazyl) radical scavenging effect was determined spectrophotometrically significant radical scavenging property was observed in crude methanol extract (ME) with IC50 of 21.95 ?g/ml. Here, butylated hydroxytoluene (BHT) and ascorbic acid (ASA) were used as standard antioxidants. The phenol and polyphenol content of the extractives varied between 56.84 to 89.58 mg of GAE/gm of dried extract. The thrombolytic and membrane stabilizing activities were assessed by using human erythrocyte and the results were compared with standard streptokinase (SK) and standard anti-inflammatory drug, acetyl salicylic acid (ASA), respectively. The extracts demonstrated significant toxicity to A. salina with LC50 values ranging from 6.07 to 7.83 ?g/ml as compared to standard Vincristine sulphate (VS, LC50 value 0.45 ?g/ml). On the other hand, the chloroform soluble fraction (CSF) of methanol extract revealed moderate antibacterial activity against some microorganisms used in the screening. Preliminary phytochemical investigation suggested the presence of flavonoids, saponins, tannins, triterpenoids and steroids. DOI: http://dx.doi.org/10.3329/bpj.v15i2.12572 Bangladesh Pharmaceutical Journal 15(2): 103-106, 2012

Evaluation of antimicrobial, antioxidant and cytotoxic activities of methanolic extracts of Lagerstroemia speciosa leaves and barks

Article history: Received on: 16/09/2012 Revised on: 08/10/2012 Accepted on: 16/10/2012 Available online: 29/10/2012 Methanolic extracts of Lagerstroemia speciosa leaves (MLL) & barks (MBL) have been evaluated for their antimicrobial, antioxidant & cytotoxic activity. Antimicrobial activity of the extracts was evaluated against 11 Gram-positive, Gram-negative bacteria and 3 fungi using disk diffusion technique. Kanamycin (30μg/disc) was used as standard. Antioxidant potentiality of the extracts was investigated on DPPH scavenging activity, Total antioxidant capacity, Reducing ability as well as total phenolic contents. Cytotoxic study was done by brine shrimp lethality bioassay and vincristin sulphate was used as standard. In antimicrobial study, the average zone of inhibition exhibited by MLL & MBL (each 500μg/disc) was 10-20 mm & 12-21mm respectively. In DPPH scavenging activity, IC50 value was found 27.89 ± 0.83μg/ml,21±0.61 μg/ml and 16.76 ± 1.11μg/ml for MLL, MBL and standard ascorbic acid, respectively. Total antioxidant capacity was found 398.37 ± 0.22 mg/g and 346.37 ± 4.02 mg/g equivalent of ascorbic acid for MLL & MBL, respectively. Reducing ability was found concentration dependent for both the extracts. The total phenolic content was found 71.06 ± 2.01 and 60.65 ± 2.16 mg/g equivalent of gallic acid for MLL & MBL, respectively. The cytotoxicity exhibited by MLL was promising with LC50 value 9.602μg/ml, comparing with the LC50 (6.25 μg/ml) values of standard vincristin sulphate as a positive control. The results suggest into the plant extracts could be used as a potential therapeutics in many pathological conditions.

In vitro antimicrobial and cytotoxicity screening of n-hexane, chloroform and ethyl acetate extracts of Lablab purpureus (L.) leaves

In the present study the antimicrobial & cytotoxic activity of crude extracts (Chloroform, n-Hexane, Ethyl acetate) of leaves of Lablab purpureus L. were studied. Antimicrobial activity was tested against eleven important pathogenic bacteria including both gram positive and gram negative bacteria and three fungi. The bacteria are B. megaterium, B. subtilis, Staphylococcus aureus, Sarcina lutea, Escherichia coli, Salmonella paratyphi, S. typhi, Shigella boydii, S. dysenteriae and Vibrio mimicus & V. parahemolyticus. Disc diffusion technique was used for invitro antibacterial and antifungal screening. Here kanamycin disc (30g /disc) was used as standard for antibacterial study. The extracts showed antimicrobial activity against most of the bacterial strains with an average zone of inhibition of 8-20mm. The tested fungi are Saccharromyces cerevaceae, Candida albicans and Aspergillus niger. The extracts showed moderate to good antifungal activity with an average 9 -15 mm zone of inhibition. Among the three solvent extracts used, the most effective extract was found to be n-Hexane extract and maximum activity (20 mm, zone of inhibition) found against Staphylococcus aureus with Minimum inhibitory concentration (MIC) values of 64g/ml. The maximum zone of inhibition for chloroform extract showed 17mm against Bacillus subtilis and E.coli with MIC 128g/ml and 32g/ml respectively. The maximum zone of inhibition for Ethyl acetate extract showed 17mm against Vibrio mimicus with MIC values of 64g/ml .Cytotoxicity test was also studied by Brine Shrimp Lethality Bioassay and compare with LC50 values of standard Vincristin sulphate as a positive control. The results illustrated significant cytotoxicity against A. salina, with LC50 13.88µg/ml, 19.17µg/ml and 17.97µg/ml for n-Hexane, Chloroform and Ethyl acetate extracts respectively.

Neurological Toxicity Profile of Weekly Vincristine Sulfate Liposomes Injection (Marqibo®) 2.25 mg/m2 Compared to Historical Standard Vincristine Sulfate 1.4 mg/m2, 2 mg, and 4 mg Administered Less Frequently

Abstract 4247 The accumulated clinical experience with standard vincristine sulfate (VCR) describes a safety profile most notable for the occurrence of a dose-related, symmetrical, sensorimotor and autonomic polyneuropathy. The various manifestations of neuropathy are the usual dose-limiting toxicities associated with standard VCR that limit the extent to which patients can derive clinical benefit from this therapy. Vincristine sulfate liposomes injection (VSLI, Marqibo®) is a sphingomyelin-based, liposomal formulation of VCR designed to intensify dose, prolong encapsulated drug circulation time, and target drug to sites of active cancer. We report here the neurological toxicity profile (Graded using CTCAE criteria v3.0) of weekly, single-agent IV VSLI 2.25 mg/m2 in 83 adult subjects with heavily pre-treated, relapsed and/or refractory acute lymphoblastic leukemia (ALL) and 100% past standard VCR exposure. Neuropathy signs and symptoms were rigorously evaluated at baseline and weekly during VSLI treatment in order to maximize adverse event detection. Findings were compared to baseline (post-standard VCR and before VSLI) neuropathy in the same VSLI treated population and to the best available published neurotoxicity data regarding standard VCR administration. In the study by Haim et al ([1] Cancer 1994; 73:2515–9), 104 VCR-naïve patients with lymphoma received multi-agent therapy including standard VCR 1.4 mg/m2 weekly to once every 28 days. In the study by Verstappen et al ([2] Neurology 2005; 64:1076–7), 114 VCR-naïve patients with lymphoma received multi-agent therapy including fixed dose standard VCR 2 mg or 4 mg every 3 weeks. Results are provided in the table below. NR = not reported. The baseline frequency of residual neuropathy in adults with ALL treated with weekly VSLI 2.25 mg/m2 was not unexpected considering that all patients received prior standard VCR. The on-study, incremental neuropathy burden was relatively modest. On-study neuropathy rates during and immediately following weekly VSLI 2.25 mg/m2 were comparable to or less than those reported in lymphoma patients receiving standard VCR at 1.4 mg/m2 every 1 to 3 weeks or at a fixed dose of 2 mg or 4 mg every 3 weeks, despite the dose intensification afforded by VSLI. Constipation, the most commonly reported neurological toxicity associated with standard VCR and VSLI, resulted in standard VCR dose reduction in 22% of patients reported by Haim et al compared to weekly VSLI dose reduction in only 5% of patients due to constipation. Frequent neurological assessments combined with a dose adjustment algorithm facilitated continued VSLI dosing in order to induce remission and minimization of Grade 3 or greater neurotoxicity. One VSLI patient developed peripheral motor neuropathy requiring drug discontinuation after 11 weekly doses. In the VSLI study, neurological toxicity was considered predictable and manageable. Disclosures: Schiller: Talon Therapeutics: Research Funding. Deitcher:Talon Therapeutics: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees.

Single-Agent Vincristine Sulfate Liposomes Injection (Marqibo®) Compared to Historical Single-Agent Therapy for Adults with Advanced, Relapsed and/or Refractory Philadelphia Chromosome Negative Acute Lymphoblastic Leukemia

Abstract 2592There are no standard of care or approved treatments specifically for advanced, relapsed and/or refractory adult Philadelphia chromosome negative (Ph-) acute lymphoblastic leukemia (ALL). Pronounced toxicity of multi-agent salvage therapy, residual toxicity in heavily pre-treated subjects, and poor response to prior multi-agent therapy including hematopoietic stem cell transplant (HSCT) may prompt the use of single-agent salvage including investigational agents. We compared the efficacy and early plus induction (30-day) mortality rates for single-agent weekly intravenous vincristine sulfate liposomes injection (VSLI, Marqibo®) 2.25 mg/m2 (no dose cap) in adults with Ph- ALL in second or greater relapse or that had progressed following 2 or more lines of anti-leukemia therapy (RALLY Study, N=65) with those from the not previously described, single-agent (non-VSLI), second salvage, Ph- ALL subpopulation (N=56) from a large published report (O’Brien et al, Cancer 2008; 113:3186–91). Despite inclusion of subjects requiring third and greater salvage therapy in the RALLY Study, the O’Brien single-agent population represents the best historical comparator to the RALLY Study population. The most common of the 28 different agents used in the O’Brien study were vinorelbine (6), clofarabine (5), nelarabine (4), and topotecan (4). No subject in the O’Brien study received single-agent standard vincristine sulfate as second salvage. The table below highlights key characteristics of the two ALL populations as well as key efficacy and toxicity outcomes.RALLY Study VSLI (N=65)O'Brien 2008 Various Agents (N=56)Age (yrs), Median (range)31 (19–83)41 (17–73)Unfavorable Cytogenetics, N (%)33 (50.7)5 (8.9)Prior Lines of Therapy, N (%)Two 32 (49.2) Three 24 (36.9) ≥ Four 9 (13.8)Two 56 (100) Three 0 ≥ Four 0Prior HSCT, N (%)31 (47.7)0Prior Standard Vincristine, N (%)65 (100)56 (100)CR+CRi+PR, N (%)19 (29.2)2 (3.6)CR+CRi, N (%)13 (20.0)2 (3.6)CR+CRi Duration (wks), Median (range)23.1 (4.6–66.1)5 and 14*Overall Survival (wks), Median (range)19.9 (1.9–94.4)7.5 (0–110)Induction (30-day) mortality, N (%)8 (12.3)17 (30.4)Overall, the RALLY Study population was more advanced and heavily pre-treated than the O'Brien population. Despite the poorer prognosis, universal prior standard vincristine exposure, and majority of subjects requiring fourth or greater line therapy, single-agent VSLI 2.25 mg/m2 (no dose cap) resulted in a higher complete response (CR+CRi) rate, complete plus partial response (PR) rate, median complete response duration (* no median was calculated for the O'Brien Population because there were only 2 responders), and overall survival duration than was observed with a variety of third-line single-agent adult Ph- ALL salvage therapies. The median overall survival durations for the RALLY Study subjects achieving CR (7), CRi (6), and PR (6) were comparable. The majority of RALLY Study complete responders had a remission duration sufficient to facilitate a subsequent, and potentially curative HSCT. In fact, 12 (18.5%) of the 65 subjects in the RALLY Study were able to undergo a post-VSLI HSCT and 5 (7.7%) subjects were long-term survivors (post-VSLI survival greater than 1 year). Single-agent VSLI was associated with a more favorable early plus induction (30-day) mortality rate in the RALLY Study than observed in relation to other single-agent therapies. Single-agent weekly VSLI 2.25 mg/m2 has the potential to provide a relatively safe and effective induction treatment and “bridge” to HSCT compared to historical non-VSLI single-agent therapies. Disclosures:Deitcher:Talon Therapeutics: related to an employee with equity ownership. Deitcher:Talon Therapeutics: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees.

Vincristine Sulfate Liposomes Injection (Marqibo®) Facilitates Durable Remissions and Potentially Curative Hematopoietic Stem Cell Transplantation in Adults with Advanced, Relapsed and/or Refractory Acute Lymphoblastic Leukemia

Abstract 4235A durable response in advanced, relapsed and/or refractory adult acute lymphoblastic leukemia (ALL) may be defined as remission that results in a meaningful prolongation of life or response that facilitates “bridging” to a subsequent, potentially curative, hematopoietic stem cell transplantation (HSCT). Vincristine sulfate liposomes injection (VSLI, Marqibo®) is a sphingomyelin/cholesterol (SM/Chol) nanoparticle formulation of standard vincristine sulfate (VCR) designed to facilitate dose intensification, prolonged drug delivery and enhanced lymphoid malignancy penetration and concentration without increased toxicity. Recently, VSLI was evaluated in a multi-institutional, Phase 1/2 (VSLI-06; NCT00144963) clinical trial and a multi-national, Phase 2 (HBS407; NCT00495079) clinical trial in a combined 101 adults (median age 31 years [range 18 to 83 years]) with advanced, relapsed and/or refractory ALL. All but 1 patient had Philadelphia chromosome negative disease. Thirteen patients (13%) had extramedullary disease, 37 (37%) had undergone a prior HSCT, and 100% had received at least one prior line of therapy including standard VCR. Study VSLI-06 (N = 36) was a dose-ascending trial of weekly VSLI (1.5 to 2.4 mg/m2) combined with pulse dexamethasone. Study HBS407 was a single-arm trial of weekly single-agent VSLI at the maximum tolerated dose established in VSLI-06 of 2.25 mg/m2. Overall, 19 (19%) patients received VSLI as a first salvage therapy, 57 (56%) patients received VSLI as a second salvage therapy, and 25 (25%) patients received VSLI as a third or greater salvage therapy. All patients had to be deemed ineligible for immediate HSCT in order to enroll in VSLI-06 or HBS407. In the combined study population, the overall response rate (complete remission [CR], CR with incomplete hematologic recovery [CRi], partial remission [PR], and bone marrow blast response [BMB]) was 31% (95% CI: 22–41) with a 20% (95% CI: 13–29) rate of CR+CRi. Despite delivering intensified individual (2.8–5.5 mg) and cumulative (up to 70.1 mg) doses of VCR, VSLI had a similar safety profile to that reported for the approved dose of standard VCR. VSLI enabled bridging to a post-VSLI HSCT in 12 of 65 (18%) patients in HBS407 and 5 of 36 (14%) patients in VSLI06 for a total of 17 of 101 (17%). All 17 post-VSLI HSCT patients were under the age of 60 years. Three of 12 post-VSLI HSCT patients from HBS407 remain alive at greater than 28, 33, and 35 months following VSLI, respectively. All 12 patients lived for greater than 100 days after post-VSLI HSCT. Long-term survival (greater than 12 months) was achieved in 27% of those able to receive post-VSLI HSCT. These outcomes, that are important to patients, may reflect the effectiveness of the VCR dose intensification facilitated by VSLI. The neuropathy associated with the dose intensified VCR administered as VSLI was predictable, manageable, and comparable to that published for standard VCR. The lack of early, pre-day 100, mortality following post-VSLI HSCT suggests that the sphingomyelin-based liposomal formulation did not adversely affect subsequent transplantation procedures. In conclusion, VSLI produced both clinically important endpoints of prolonged survival and achievement of response allowing for a bridge to HSCT for advanced, relapsed and/or refractory ALL. Disclosures:Schiller:Talon Therapeutics: Research Funding. Silverman:Talon Therapeutics: Employment, Equity Ownership. Deitcher:Talon Therapeutics: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees.

Antidiabetic, cytotoxic activities and phytochemical screening of Peltophorum pterocarpum (DC.) K. Heyne root

model. The extract was given intraperitoneally at a dose of 200 and 300 mg/kg body weight and the hypoglycemic effects was compared with standard drug vincristine sulphate, a standard drug. The extract reduced blood glucose level in both diabetic model and at 300 mg/kg dose, the test sample showed better activity. The extract was also subjected to brine shrimp lethality bioassay and phytochemical screening test. LC 50 value of the extract was 28.25 µg/ml and for standard drug vincristine sulphate it was 14.55 µg/ml. Preliminary phytochemical screening of the extract revealed the presence of different types of compounds including flavonoids and steroids. Our results suggest that root of P. pterocarpum demonstrated promising antidiabetic activity with low cytotoxicity that substantiated its ethnomedicinal use and may provide new molecules for the treatment of diabetes.

Antibacterial, cytotoxic and antioxidant activity of chloroform, n-hexane and ethyl acetate extract of plant Coccinia cordifolia

The main aim of this study was to find out the antibacterial, cytotoxic and antioxidant activity of chloroform, n-hexane and ethyl acetate extracts of Coccinia cordifolia (Family: Cucurbitaceae). Disc diffusion technique was used for in vitro antibacterial screening against gram positive and gram negative human pathogenic bacteria. Here kanamycin disc (30g/disc) was used as standard. The chloroform extract of Coccinia cordifolia showed good antibacterial activity with the average zone of inhibition 9-12mm. The n-hexane and ethyl acetate extract showed average zone of inhibition 7-10mm and 7-11mm respectively. The Brine shrimp lethality bioassay method was used to determine the cytotoxic activity and vincristine sulphate was used as positive control. The LC50 values of standard vincristine sulphate, chloroform, n-hexane and ethyl acetate extract were 7.55μg/ml, 23.96 μg/ml, 14.12 μg/ml & 15.49 μg/ml respectively which indicate the plants extracts compounds are promisingly cytotoxic and they might have antitumer and pesticidal activity. Among the extracts the chloroform extract showed the highest cytotoxic activity with LC50 23.96 μg/ml. Antioxidant activity test of the crude extracts were assessed by means of DPPH free radical scavenging method where ascorbic acid was used as standard with IC50 43.22 μg/ml. The ethyl acetate fraction of Coccinia cordifolia showed strongest antioxidant activity with IC50 value of 50.98 μg/ml.