Absence of antibiotic activities of Cenchrus setigerus and cenchrus ciliaris seed extracts in different polar solvents

Premlata Singariya, Padma Kumar, Krishan K Mourya

Journal of Pharmaceutical Negative Results 2013 4(1):71-75

Aim: The aim of present study is to investigate the antibiotic activity of seeds of Cenchrus setigerus and Cenchrus ciliaris extracts, in order to use them as a possible source of new anti-microbial substances against important human pathogens. Materials and Methods: Crude extracts of seeds of both species of Cenchrus grass were evaluated against some important G-ve bacteria. Escherichia coli, Raoultella planticola, Enterobacter aerogenes and one fungus Aspergillus flavus. The dried and powdered seeds were successively extracted with hexane, toluene, isopropyl alcohol, acetone and ethanol using the soxhlet assembly. The antimicrobial activity assay was done by both disc diffusion and serial dilution methods. Results: The results indicate that all the extracts, in different polar solvents did not show any antibacterial activity against R. planticola or any antifungal activity against A. flavus. Conclusion: All extracts in the different polar solvents did not have or had very less antibacterial and antifungal activities.

Evaluation of drug promotional literatures using WHO guidelines

Tejas Khakhkhar, Maulin Mehta, Rima Shah, Dineshchandra Sharma

Journal of Pharmaceutical Negative Results 2013 4(1):33-38

Aim: To evaluate the scientific and ethical status of the drug promotional literatures available in Indian market using WHO criteria. Materials and Methods: A cross-sectional observational study was carried out in department of pharmacology for evaluation of 142 drug promotional literatures by WHO-criteria, collected randomly from various regions of Gujarat. They were also analyzed for different claims, catchy terms, quality of paper and print, and representation of data with statistics/diagram/table. The references cited in the literatures were evaluated for their source, year of publication, authenticity, and retrievability. Results: 49% of literatures were designed for promotion of fixed dose-drug combinations (FDCs). Chemotherapeutic agents and cardiovascular drugs were most promoted drug groups (19% each). None of the drug promotional literature fulfilled all the WHO criteria. Description of pharmacological effects and mechanism of action was not given in 54% and 80% of literatures, respectively. Majority (80-90%) were lacking information related to indications, correct dosage regimen, and dose adjustments in special situations. Most neglected aspect of drug promotion was mentioning about adverse drug reactions, drug interactions, precautions, and over dosage (<10%). False/tall claims, catchy/broken statements were given in 86% and 72% of literatures, respectively. Irrelevant diagrams were shown in 69%, statistical data for support in 7%, and tabular presentation in 5% of literatures. References were cited in 67% of literatures, of which 98% were from indexed-journals and were retrievable. Conclusion: Critical review of drug promotional literatures can make drug prescribing more effective. If drug promotional literatures fulfill all WHO guidelines, it can make promotion ethical and rational.

In vitro pharmacological investigations of Biophytum sensitivum callus extract: Lack of potent activities

Sirigiri Chandrakala, Kokkanti Mallikarjuna, Challa Siva Reddy

Journal of Pharmaceutical Negative Results 2013 4(1):60-65

Objective: Biophytum sensitivum is an important medicinal plant extensively used in traditional oriental herbal medicines. Though medicinal use of this plant is known, the active principles responsible such property is not known. Pharmacological screening of this plant may lead to discovery of new activity with new mode of treatment. Hence, screening for in-vitro pharmacological activities of methanolic callus extracts of Biophytum sensitivum has been carried out. Experiments were designed according to the standard methods and processes. Materials and Methods: Leaf cutting derived callus on MS medium supplemented with BA (Benzyl adenine) 1.0 mg/l + NAA (1-naphthaleneacetic acid) 1.0 mg/l is used as a source, and compounds were extracted from dried callus using methanol solvent with Soxhlet apparatus. Results: The callus extract has shown antioxidant activity, in-vitro inhibition of enzyme activities like &#945;-glycosidase, acetyl cholesterase, and tyrosinase, but potency was found to be low. The Graph pad Prism Version-5 software is used to analyze data in the form of Figures. Conclusion: For the first time, we are reporting in-vitro pharmacological screening of methanolic callus extracts of Biophytum sensitivum.

Lack of the cytochrome P450 3A interaction of methanolic extract of Withania somnifera, Withaferin A, Withanolide A and Withanoside IV

Jay Savai, Alice Varghese, Nancy Pandita

Journal of Pharmaceutical Negative Results 2013 4(1):26-32

Aims: Withania somnifera is widely employed as a rejuvenator and expected to promote physical health and increase longevity. The aim of the present research work was to evaluate Cytochrome P450 3A (CYP3A) interaction of Withania somnifera. Materials and Methods: In vitro CYP3A interaction of methanolic extract of Withania somnifera (WS) and its principal phytoconstituents: Withaferin-A (WA), Withanolide-A (WL-A) and Withanoside-IV (WS-IV) were investigated in rat and human liver microsomes. In vivo CYP3A interaction potential was investigated by administering methanolic extract of WS orally at a dose of 500 mg/kg in female Wistar rats. Sildenafil citrate was used to index the activity of CYP3A. Results: IC 50 values of methanolic extract of Withania somnifera, WA, WL-A, WS-IV were found to be 200 &#956;g/ml, >20 &#956;M, >64 &#956;M and >64 &#956;M for CYP3A both in rats and humans, respectively. When sildenafil citrate was orally co-administered with methanolic extract of WS and compared with orally administered sildenafil citrate alone, the area under plasma concentration time (AUC) curve and C max did not significantly differ as compared to the group which received rifampicin orally (positive control). Conclusions: Results suggested that methanolic extract of WS, WA, WL-A, WS-IV showed no in vitro CYP3A inhibition in rats and humans. Methanolic extract of WS did not significantly alter the pharmacokinetics of sildenafil citrate in rats; indicating its safety when co-administered with other drugs that are substrates of CYP3A. Thus the results indicate the lesser likelihood of drug herb interactions when concomitantly administered with CYP3A substrates.

Effect of rimonabant on the components of metabolic syndrome: A randomized, controlled study done on Punjabi population

Rupinderjeet Kaur, Paramdeep Singh, Amarpreet Kaur, Devinder S Mahajan, Harharpreet Kaur

Journal of Pharmaceutical Negative Results 2013 4(1):46-53

The objectives of this study were to study the total and differential effect of Rimonabant on weight parameters, lipid profile, glycemic profile, and blood pressure and to evaluate its safety profile in obese population of Punjab. Materials and Methods: This was a randomized open controlled study, carried out on 100 subjects that included Punjabi adults (18-70 years) who were obese or overweight (with associated risk factors) according to Asia pacific guidelines. Treatment group (n = 50) was given Rimonabant 20 mg once-daily, and control group (n = 50) was given placebo for 12 weeks. Alterations in weight and lipid parameters, Fasting blood sugar (FBS), and blood pressure were recorded. Psychiatric adverse events were monitored by HADS score. Analysis was done using SPSS software. Results: Statistically significant difference was observed in two groups in reduction in weight, waist circumference, body mass index. Overall reduction in number of patients in the obese category was 20% in group 1 and 6.25% in group 2. In lipid parameters, significant changes were observed in triglycerides (TG), HDL cholesterol, and total cholesterol: HDL ratio (P < 0.001). No significant change was observed for LDL cholesterol and total cholesterol. Effect on Fasting Blood Sugar (FBS) was also significant. Moderate decrease in systolic blood pressure and no significant change was observed in diastolic blood pressure. Using multivariate regression model, Rimonabant was found to have weight-loss independent effect on lipid parameters with triglycerides having the maximum percentage difference between regression coefficients. No significant correlation was observed between changes in lipid parameters and weight loss. Amongst the lipids, changes in triglycerides and HDL cholesterol correlated best. Decrease in FBS correlated with weight loss but not to lipid parameters. Adverse event profile was comparable in both groups. Depressive symptoms were noted in 3 subjects in treatment group with 1 requiring termination of treatment. Frequency of anxiety symptoms was same in both groups with none showing tendency towards suicide. Conclusions: This is the first study of Rimonabant on Punjabi population and reports the depressive adverse events to be occurring in low frequency and of mild intensity in this group. Thus, it provides a clue to the need for further studies of cannabinoid receptor antagonists on larger samples in such ethnically predisposed populations to reevaluate the safety profile and its genetic correlation. It also suggests need for further research on differential effect of antagonism of cannabinoid receptors in CNS and adipose tissue and its use to produce drug that targets metabolic derangements by selective action on adipose tissue receptors.

Insignificant level of in vitro cytotoxicity, anti-rotavirus, antibacterial, and antifungal activities of N-alkylmaleamic acids

VJ Belinelo, M.S. Tacchi Campos, RM Antunes, R.A.G. Assenco, S.A. Vieira Filho, M.C.S. Lanna, EC Marçal, T.H.S. Fonseca, MA Gomes, JC Magalhães

Journal of Pharmaceutical Negative Results 2013 4(1):19-25

By reacting maleic anhydride with amines, we synthesized the derivatives N-ethyl, N-(2-ethylamine), N-piperidinyl, N-phenyl, and N-phenylhydrazinyl maleamic acids. The purity of these products was initially verified by melting range and the presence of only one spot observed by thin layer chromatography. The chemical structures of the obtained N-alkyl maleamic acids were confirmed through infrared (IR) and hydrogen and carbon nuclear magnetic resonance ( 1 H and 13 C NMR) spectrometry. Due to the already proven pharmacological activity of maleimides, maleic anhydride and its N-alkyl maleamic acids were subjected to in vitro assays to observe antiviral (SA-11 rotavirus), antibacterial (Escherichia coli, Staphylococcus aureus, and Bacillus cereus), antifungal (Colletotrichum musae, Fusarium solani f. sp. phaseoli, Fusarium solani f. sp. piperis Alb., and Penicillium sp.), and antiprotozoal (Trichomonas vaginalis, Giardia lamblia, and Entamoeba histolytica) effects. To study the anti-rotavirus properties, firstly the 3-(4,5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide (MTT) method was used to establish the median cytotoxicity concentration (CC 50 ) of the compounds, using MA-104 cell line. Under the experimental conditions used, cytotoxic, anti-rotavirus, antibacterial, and antifungal properties were not observed for these compounds.

Lack of brine shrimp lethality and hemagglutination activity in Grewia asiatica Linn

Abidah Parveen, Mohammad Irfan, Muhammad Saleem Jilani, Kashif Wasim, Mehwish Kiran, Fida Muhammad

Journal of Pharmaceutical Negative Results 2013 4(1):1-4

Objective: To investigate the brine shrimp lethality and hemagglutination activity of the fruit, stem bark and leaves of Grewia asiatica L. (Family: Tiliaceae). Materials and Methods: The crude ethanolic extracts and fractions of the fruit, stem bark, and leaves were subjected to cytotoxic assay using brine shrimps and were also investigated for hemagglutination activity. Results: Brine shrimps survived easily and hemagglutination activity was not observed. Conclusions: The fruits, leaves and stem bark of G. asiatica have insignificant brine shrimp lethality and hemagglutination activity was found to be absent.

Lack of human immunodeficiency virus-1 integrase inhibitory activity of novel 3a, 4, 7, 7a-tetrahydro-1H-isoindole-1,3 (2H)-dione derivatives

Ashok Penta, Kakamanu Kishore Babu, Swastika Ganguly, Sankaranarayanan Murugesan

Journal of Pharmaceutical Negative Results 2013 4(1):13-18

Background: Majority of reported integrase (IN) inhibitors had an important structural feature, i.e., 1,3-diketo functional group. It plays a vital role in IN inhibition by the formation of chelating triod with Mg +2 ions. Materials and Methods: A novel series of fifteen 3-(1,3-dioxo-3a, 4-dihydro-1H-isoindol-2 (3H,7H,7aH)-yl)-N-(substituted phenyl) propanamide 4(a-o) analogs were synthesized by reacting the corresponding 3-chloro-N-(substituted phenyl) propanamides 2(a-o) with 3a, 4,7,7a-tetrahydro-1H-isoindole-1,3 (2H)-dione (3) in acetonitrile medium in the presence of potassium carbonate. Various substituted 3-chloro-N-(substituted phenyl) propanamides 2(a-o) were synthesized by treating appropriate substituted anilines 1(a-o) with 3-chloro propionyl chloride in dichloromethane as solvent in the presence of triethylamine as base. The synthesized compounds have been characterized on the basis of fourier transform infrared spectrophotometerproton nuclear magnetic resonance spectrophotometer, 1 H NMR, Mass spectral and Elemental Analysis. Results: All the synthesized compounds were evaluated for their human immunodeficiency virus (HIV)-1 INinhibitory activity. However, unlike other anti-IN agents, none of these molecules showed inhibition of either 3' processing, and strand transfer reactions of HIV-1 IN.

Chronic use of 17β-Ethinyl estradiol on cardiovascular hemodynamic profile: "Friend or foe"?

Hira Lal Bhalla, Mandeep Kumar Arora, KK Saxena, William R Surin

Journal of Pharmaceutical Negative Results 2013 4(1):54-59

Introduction: The effects of ovariectomy (Ovx), menopause, and estrogen replacement on the hemodynamic remain controversial. This study employed the technique of impedance cardiography analysis to measure the effect of chronic use of estrogen replacement on cardiovascular hemodynamic in the Ovx rats. Materials and Methods: Colony-bred adult Ovx female Sprague-Dawley rats were randomized into three groups: 17 &#946;-Ethinyl Estradiol treated ovariectomized group (OvxE), vehicle treated ovariectomized group (OvxV), and Sham Operated (SO). Animals received 17 &#946;-Ethinyl Estradiol (17 &#946;-EE) once daily for 90 days. Cardiovascular hemodynamic parameters such as left ventricular ejection time (LVET), pre-ejection period (PEP), Systolic time interval (STI), cardiac output (CO), cardiac index (CI), stroke volume (SV), and stroke volume index (SVI) were assessed 24 h after last treatment on 7, 15, 30, 60, or 90 days. Results: Compared to SO group, Ovx with or without estrogen replacement did not significantly affect the mean blood pressure, CO, CI, SV, and SVI. No significant changes were observed in LVET and PEP from SO. Treatment with estradiol increased the STI by 66.62% and 53.60% ( P < 0.05), from control after 60 and 90 days, respectively. Blood velocity, base impedance (Zo) and maximum change in impedance during systole (Zt) corresponding to time-varying fluid volume (blood) remained within normal limits of variation. Conclusion: These results demonstrate that on long-term administration of estrogen significantly increased STI in rats.

Treatment with N-acetylcysteine does not alter blood glucose levels and the oxidative stress status in diabetic rats

André Valle de Bairros, Miguel Roehrs, Gianine Ribeiro, Fernando de Freitas, Ana Paula Moreira, Raquel Tonello, Cinthia Mazzanti, Mirna Leal, Vera Maria Morsch

Journal of Pharmaceutical Negative Results 2013 4(1):5-12

Objectives: To verify the contribution of N-acetylcysteine (NAC) as an antioxidant drug in the therapy of diabetes, helping to reduce the deleterious effects resulting from oxidative stress associated with the hyperglycemic state. Materials and Methods: The animals were divided into normal (saline, 25 mg/kg NAC, and 75 mg/kg NAC) and diabetic rats (saline, 25 mg/kg NAC, and 75 mg/kg NAC) with five rats per group, and were treated or four weeks. Diabetes induction was performed by intraperitoneal injection of alloxan after fasting for 12 hours. Subsequently, glucose solution was used to promote wear of the pancreatic beta cells. Blood parameters such as glucose, glycated hemoglobin, hepatic and renal biomarkers, and butyrylcholinesterase activity were determined by commercial kits. Catalase, glutathione peroxidase, and superoxide dismutase activities were measured using spectrophotometric techniques, while glutathione and malondialdehyde levels were determined by chromatographic techniques. Results: NAC had no significant differences on glycemic, hepatic, renal, and oxidative stress biomarkers. Superoxide dismutase activity was significantly higher ( P < 0.05) in diabetic rats treated with NAC compared to the diabetic saline group, while butyrylcholinesterase activity was significantly lower ( P < 0.05) in the same groups. There was a negative correlation between superoxide dismutase and butyrylcholinesterase activities. Conclusion: NAC supplementation did not re-establish the antioxidant system and consequently the deleterious effects of diabetes did not decrease. Diabetic groups that received NAC demonstrated that superoxide dismutase activity was indirectly linked to the levels of butyrylcholinesterase. More studies are necessary to investigate the action of NAC on superoxide dismutase and butyrylcholinesterase activities in the diabetic state.

Insignificant effect of cleistanthin A and cleistanthin B on motor function in animal models

Chavan Madhavrao, Subramani Parasuraman, Ramasamy Raveendran

Journal of Pharmaceutical Negative Results 2013 4(1):66-70

Aim: To study the in vitro and in vivo effects of cleistanthin A and cleistanthin B on the cholinergic receptors and the motor function using mouse as an animal model. Materials and Methods: The cleistanthin A and cleistanthin B were isolated from the leaves of Cleistanthus collinus using column chromatography and purified. The effect of cleistanthins A and B on locomotor activity, motor co-ordination and grip strength in mice were evaluated using actophotometer, rota-rod apparatus and grip strength test respectively (in vivo). The effect of cleistanthins A and B on nicotinic acetylcholine receptor was studied using a mouse esophagus preparation (in vitro) and the concentration-response curves of carbachol and cleistanthins A and B were recorded using a data acquisition system (Biopac Inc. USA) through a variable transducer (500 g). Results: Cleistanthins A and B did not have any significant effect on locomotor activity, motor co-ordination and grip strength when they were used upto 400 mg/kg BW. Cleistanthins A and B did not show any significant inhibitory effect on nicotinic acetylcholine receptors in isolated mouse esophagus tissue preparation at 1, 3, and 10 &#956;g doses. Conclusion: Cleistanthins A and B do not have significant activity on the motor system when used upto 400 mg/kg in mice and also they did not have any effect on nicotinic acetylcholine receptors when used upto 10 &#956;g in the isolated mouse esophagus preparation.

Absence of antidiabetic activity in some novel thiazolidinone derivatives

Tejprakash Singh, Pramod Kumar Sharma, Nitin Kumar, Rupesh Dudhe

Journal of Pharmaceutical Negative Results 2013 4(1):39-45

Aim: It was aimed to synthesise some novel thiazolidinone derivatives and assess them for antidiabetic activity. Material and Methods: A series of substituted 5-ethylidene-2-(phenylimino) thiazolidin-4-ones were prepared by using phenylthiourea (I) as a starting material. Phenylthiourea on reaction with ethylchloroacetate, in the presence of ethanol and fused sodium acetate, gave 2-(phenylimino) thiazolidin-4-one (II), and 2-(phenylimino) thiazolidin-4-one on further reaction with substituted benzaldehyde gave substituted 5-ethylidene-2- (phenylimino) thiazolidin-4-one (III – XVIII). The synthesized compounds were authenticated on the basis of elemental analysis, IR, 1H NMR, and Mass spectral analysis and some of the compounds were selected on the basis of a literature review, to evaluate them for their antidiabetic activity. Results and Conclusion: All The tested compounds 5-(4-fluorobenzylidene)-2-(phenylimino) thiazolidin-4-on (VII) and 5-(4-Methylbenzylidene)-2-(phenylimino)thiazolidin-4-one (X), 5-(2, 4-dinitrobenzylidene)-2-(phenylamino) thiazolidin-4-one (XVII) were found to be ineffective in lowering the blood glucose level.

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