Design and one-pot synthesis of new α-aminophosphonates and antimicrobial activity

Zahra Rezaei, Soghra Khabnadideh, Younes Ghasemi, Masomeh Fadaei, Zeinab Karimi

Journal of Pharmaceutical Negative Results 2011 2(2):78-86

Background: α-Aminophosphonates are bioisosters of amino acids and have several pharmacological effects. α-Aminophosphonates have been synthesized by various routes from reaction between an amine, an aldehyde, and phosphate compounds.
Materials and Methods: We synthesized 20 new α-aminophosphonates in the presence of FeCl3 in THF as a catalyst to facilitate the Manich-type reaction of aldehyde, amine and phosphite compounds to form the corresponding α-aminophosphonates in a one-pot, three-component reaction. In this study, the catalytic effect of ZnCl2 was also compared with FeCl3 in the synthesis of α-aminophosphonates. Results: The results showed that FeCl3 catalyzed the reaction in mild conditions to form α-aminophosphonates with high yields, but ZnCl2 did not give high yields of the compounds and the reaction took longer time in comparison to that taken by FeCl3 The chemical structures of all new compounds were confirmed by spectrophotometric methods (1HNMR, 13CNMR, IR). The compounds were investigated for antimicrobial activity against Escherichia coli, Bacillus subtilis, Salmonella typhi, Shigella sonnei, Proteus vulgaris, and Staphylococcus epidermidis. Conclusion: The new synthesized compounds did not show good antibacterial activity against the tested microorganisms

Insignificant difference between biofilm forming isolates of filtered water and non-filtered water

Sangita Revdiwala, Summaiya Mulla, Nrupal Chevli

Journal of Pharmaceutical Negative Results 2011 2(2):110-114

Aim: Biofilm formation is a developmental process with intercellular signals that regulate growth. Biofilms contaminate catheters, ventilators, and medical implants; they act as a source of disease for humans, animals, and plants. In this study we have done a quantitative assessment of biofilm formation in bacterial isolates, associated with the drinking water distribution system, in tryptic soya broth, with different incubation times. Materials and Methods: The study was carried out on 104 samples of water from different systems. The bacterial isolates were processed as per the microtiter plate method with only tryptic soya broth, and with varying concentrations of glucose, and were observed in response to time. Results: Out of the total of 104 samples of water, 63 were found to be bacteriologically positive and 24 of them were biofilm formers. Enterobacter spp. and Pseudomonas were maximally found to be biofilm producing. Conclusion: Biofilm formation depends on adherence of bacteria to various surfaces. Overhead plastic tanks and metal taps were the most common sources being infected. We must have a regular policy to get them clean so as to have affordable and safe drinking water.

Antacids incorporation in immediate release tablets failed to improve the stability of omeprazole in acidic media

HV Chavda, TM Chaudhary, CN Patel

Journal of Pharmaceutical Negative Results 2011 2(2):73-77

Background: The stability of omeprazole (OMZ) decreases in acidic medium. In this investigation, attempts have been made to develop oral tablet containing antacids/buffers to increase the pH of dissolution media for certain time. Materials and Methods: Tablet formulations were prepared by the direct compression technique. For the selection of superdisintegrant, Croscarmellose sodium, used initially was replaced with other superdisintegrants. The prepared tablets were evaluated for hardness, weight variation, thickness, friability, drug content, disintegration time and in vitro drug release studies. During the in vitro drug release studies, the pH of dissolution media was measured. Results and Discussion: All batches showed very short disintegration time, within 0.5-2 min except F1 and S5. Batch F7 was able to provide the immediate drug release. The study showed that the incorporation of antacid improved the pH of dissolution media, but failed to maintain it. Even though the high quantities of antacids were incorporated; the stability of drug in media was not improved. Other superdisintegrants did not show any significant changes in drug release or disintegration time. Bath F7 was stable for the period of 6 months at 40 o C / 75 %RH. Conclusions: Incorporation of higher quantities of antacids failed to retain the stability of OMZ in acidic media.

Sodium nitrite therapy fails to improve tissue perfusion in a mouse model of hind limb ischemia: Slight differences in methodology may be responsible casting suspicion on the reliability and predictive value of this model

Jeff S McKee, Benjamin D Brooks

Journal of Pharmaceutical Negative Results 2011 2(2):99-106

Background: The nude mouse model of hind limb ischemia is used to evaluate human-derived, cell-based therapeutics intended to promote tissue perfusion. The criticism of the mouse model of hind limb ischemia is the absence of a well-characterized positive control. The suitability of sodium nitrite (NaNO 2 ) was evaluated. The rationale for doing so was based on a report that NaNO 2 induced unprecedented tissue perfusion in wild-type mice using a similar model. The objective was to evaluate NaNO 2 to improve tissue perfusion in nude mice as well as their wild-type counterparts. Materials and Methods: The mice underwent surgically induced, unilateral hind limb ischemia, and received either NaNO 2 or a vehicle intraperitoneally, twice daily, for seven days. Hind limb tissue perfusion was evaluated on days one, four, seven, and fourteen post-surgery. Results: No increase in tissue perfusion was observed in the nude or wild-type mice treated with NaNO 2 when compared with the vehicle. Nude mice exhibited significantly lower tissue perfusion compared to wild-type mice, irrespective of the treatment. Conclusions: NaNO 2 failed to increase tissue perfusion and, therefore, did not appear suitable for use as a positive control in this model. This is in stark contrast to a previous report indicating that NaNO 2 significantly increased tissue perfusion in wild-type mice using a similar model. The exact cause is not known, but is probably due to differences in methodology employed between laboratories. The lower tissue perfusion in nude mice is a novel finding, suggesting this strain may have less pre-existing collateral vessels and/or a reduced capacity to form new vessels as compared to wild-type mice.

Effort toward the single pot synthesis of 6-(Biphenyl-4-yl)-2-[2-(indol-1-yl)ethyl]-4,5-dihydropyridazin-3(2H)-one from 6-(Biphenyl-4-yl)-2-[2-(indol-1-yl)ethyl]-4,5-dihydropyridazin-3(2H)-one

Mohammad Asif, Anita Singh, Lakshmayya

Journal of Pharmaceutical Negative Results 2011 2(2):69-72

Background: Many synthetic heterocyclic compounds are known to have different biological activities. Pyridazine and its derivatives are the important six membered heterocyclic compounds containing two N-hetero atoms. The pyridazine moiety is an important structural feature of many biologically active compounds. Pyridazine derivatives show diverse pharmacological properties. Aims and Objectives of the Study: The goal of this study, Pyridazine hold considerable interest relative to the preparation of organic intermediates and physiologically active compounds also. On the basis of its important in reported literature, we synthesized the 6-(Biphenyl-4-yl)-2-[2-(indol-1-yl)ethyl]-4,5-dihydropyridazin-3(2H)-one from 6-(Biphenyl-4-yl)-2-[2-(indol-1-yl)ethyl]-4,5-dihydro pyridazin-3(2H)-one in single step or pot reaction. Results : The important Observations and concrete implications of the study. The result showed that the compound 6-(Biphenyl-4-yl)-2-[2-(indol-1-yl) ethyl]-4,5-dihydropyridazin-3(2H)-one was not prepared by this method. The synthesis of compound has been confirmed by the spectral analysis namely IR, 1H NMR and Mass spectroscopy.

Reporting of sample size and power in negative clinical trials published in Indian medical journals

Jaykaran , Preeti Yadav, ND Kantharia

Journal of Pharmaceutical Negative Results 2011 2(2):87-90

Background and Aim: It is observed that negative clinical trials published in medical journals are poor in reporting of sample size calculation, various components of calculation of sample size. It is also observed that they are underpowered to detect the actual difference between the treatment outcomes. Because of scarcity of these data for Indian medical journals we designed this study to critically analyze the Indian medical journals for reporting of sample size, components of sample size and power. We calculated post hoc power for 30% and 50% difference between the treatment outcomes. Materials and Methods: All the negative clinical trials published in five Indian medical journals (Indian Journal of Pharmacology (IJP), Indian Pediatrics (IP), Indian Journal of Dermatology (IJD), Indian Journal of Dermatology, Vanereology and Leprology (IJDVL), and Journal of Postgraduate Medicine (JPGM)), between 2001 and 2008, were analyzed by each author for reporting of the sample size and components of the sample size. Post hoc power for 30% and 50% differences between the outcome was calculated by G Power software. All data were expressed as frequency, percentages, and 95% confidence interval around the percentages with the help of SPSS ver. 17. Results: The median sample size was observed to be 33 (range 9-85). Power was calculated in 28 (41.1%, 95% CI 30.2% to 53%) trials. The sample size was not calculated in 34 (50%, 95% CI 38.4% to 61.5%) trials. The full sample size was calculated in only 2 (2.9%, 95% CI 0.8% to 10.1%) trials. Post hoc power above 80% for 30% of difference was reported in 3 (4.4%, 95% CI 1.5% to 12%) trials and for 50% difference in outcome it was reported in 42 (61.7%, 95% CI 49.8% to 72.3) trials. Conclusion: Negative clinical trials published in five Indian journals are poor in reporting of sample size calculation. It is also observed that most of trials are underpowered to see the 30% and 50% difference between the outcomes. There is a need to generate more awareness regarding the sample size and power calculation

Insignificant anticancer activity of novel substituted pyrimidine derivatives

Anshu Chaudhary, Pramod Kumar Sharma, Shiv Jee Kashyap, Jitendra Kumar Gupta, Rupesh Dudhe, Prabhakar Verma

Journal of Pharmaceutical Negative Results 2011 2(2):62-68

Background: Pyrimidine and fused pyrimidine derivatives play an important role in therapeutic strategies. It is known to be most prominent structures found in nucleic acid, including uracil, thymine, cytosine, adenine, and guanine, which are fundamental building blocks for deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). Materials and Methods: A series of 3-[2-amino-6-(substituted)-pyrimidin-4-yl]-6-(substituted)-2H-chromen-2-one derivatives were prepared by reacting salicylaldehyde with ethylacetoacetate in the presence of piperidine by Knoevenagel reaction as a starting material. The chemical structures were confirmed by means of FTIR (Fourier Transform InfraRed Spectrophotometer-8400S), 1H NMR, and elemental analysis. The data of these synthesized compounds were submitted to National Institute of Health, USA, under the drug discovery program of NCI (National Cancer Institute) and screened for anticancer activity at a single high dose (10−5 M) in full NCI 60 cell lines. Results: Unfortunately, the selected compounds have not shown any potent significant anticancer activity in the NCI 60 cell line screening. Conclusion: The compound (T2) found to be most efficient anticancer activity with selective influence on breast cancer cell lines, especially on MCF7 cell line with a growth percentage of 33.63.

Ultra violet spectrophotometric method: Not possible for the simultaneous estimation of alpha one adrenoreceptor blockers

Alankar Shrivastava, Vipin Bihari Gupta

Journal of Pharmaceutical Negative Results 2011 2(2):115-120

Background: Although initially introduced for the management of hypertension, al-adrenergic-receptor antagonists (a 1 blockers) have become the standard of care for the medical management of benign prostatic hyperplasia (BPH)-related lower urinary tract symptoms (LUTS). Alpha-blockers (alfuzosin, tamsulosin, doxazosin, prazosin, and terazosin) relax the smooth muscles in the prostate and are indicated for the symptomatic treatment of BPH, due to evidence of their positive and rapid effect on LUTS. Objective: Our objective was to develop and validate a method for simultaneous estimation of these drugs. Our aim was to develop a UV spectrophotometric method because of the simplicity and economical advantage of this technique. Materials and Methods: All these drugs were dissolved in the same solvent (methanol) and scanned under a ShimadzuUV spectrophotomer, under full UV (ultraviolet) range (200 – 400 nm). Result: After some experiments we found that this is impossible. This was evident from our study that UV spectra were overlapping each other, except in the case of tamsulosin. Conclusion: Same class of drugs may have almost the same functional groups, and gradient Reverse-Phase Liquid Chromatography(RPLC) will be more useful to separate such complicated mixtures. Therefore, we propose to develop a gradient High-Performance Liquid Chromatography(HPLC) method as it may be a more suitable method for the simultaneous estimation of these drugs.

Hydroethanolic leaf extract of Ficus religiosa lacks anticonvulsant activity in acute electro and chemo convulsion mice models

Damanpreet Singh, Bikram Singh, Rajesh Kumar Goel

Journal of Pharmaceutical Negative Results 2011 2(2):58-61

Introduction: Ficus religiosa L. (Moraceae) has been of great medicinal value in traditional medicine and implicated in a wide variety of human and animal disorders. Its leaves have been used for the ethnomedical treatment of epilepsy. But its traditional antiepileptic use is not fully understood experimentally. Hence the present study was undertaken to explore the anticonvulsant effect of the leaves in experimental animal models of convulsion. Materials and Methods: The anticonvulsant effect of hydroethanolic leaf extract of F. religiosa was studied at 100, 250 and 500 mg/kg; intraperitoneally (i.p.) in maximal electroshock (MES), and at 100, 250, 500 and 600 mg/kg; i.p. doses in pentylenetetrazol (PTZ) test in mice. The duration of tonic hind limb extension(s) and latency to clonic convulsions (min) was noted in MES and PTZ tests, respectively. Phenytoin (25 mg/kg; i.p.) and diazepam (5 mg/kg; i.p.) served as reference standards in MES and PTZ tests, respectively. Percentage mortality was also noted. Results: There was no significant change observed after the extract treatment on the duration of tonic hind limb extension in MES test, and latency to clonic convulsions in PTZ test, as compared to their respective controls. Moreover, percentage mortality remained unaltered after the extract treatment. Conclusions: From the results of present study it is concluded that the hydroethanolic leaf extract of F. religiosa lacks anticonvulsant activity in MES- and PTZ-induced convulsion tests. Further studies are required from other regions and using different animal models to support these findings.

A comparative study of pheniramine and lorazepam for physiological and cognitive/psychomotor task impairment

Yatan Pal Singh Balhara, Raka Jain, Anju Dhawan, Manju Mehta

Journal of Pharmaceutical Negative Results 2011 2(2):91-98

Context: Pheniramine is being used harmfully in combination with opiates and benzodiazepines through injecting route. Aims: The present study is an attempt to compare the physiological and psychomotor/cognitive task performance on pheniramine and lorazepam. Settings and Design: The study used a double blind randomly allotted cross-over design. Materials and Methods: The doses of the drugs used were placebo (normal saline) – 2 ml, Pheniramine maleate – 45.5 mg, Lorazepam – 2 mg. The assessments were made at base line and then at 15 min., 120 min and 240 min. The subjects were assessed for the socio-demographic profile, drug use history, physiological parameters (pulse rate, BP, respiratory rate), and psychomotor/cognitive tasks. Statistical Analysis used : Analysis was carried out using SPSS ver 10.0. In between, drug comparisons were done using one-way ANOVA (multiple comparisons). Results: Physiological and cognitive/psychomotor tasks performance did not show any significant difference between pheniramine, lorazepam and placebo. Conclusions: The findings suggest the pheniramine and lorazepam have comparable impairment on physiological and cognitive/psychomotor task performance.

Lack of antiglycation activity of fresh juice of whole plant of Enicostema axillare (Lam.) Raynal

WD Ratnasooriya, K.I.W.K Somarathna, G.A.S Premakumara, E.R.H.S.S Ediriweera

Journal of Pharmaceutical Negative Results 2011 2(2):55-57

Objective: To investigate the antiglycation potential of fresh juice of whole plant of Enicostema axillare (Lam.) Raynal. in vitro. Materials and Methods: Antiglycation activity of fresh juice of whole plant was determined in vitro using five concentrations (25, 50, 100, 200, and 400 μg/ml) by determining their ability to inhibit the formation of advanced glycation endproducts in a bovine serum albumin / glucose system using fluorescence spectroscopy. Results: There is no antiglycation activity in vitro of fresh juice of E. axillare plant. Conclusion: It is unlikely that impairment of diabetic complications by E. axillare whole plant juice as claimed by ayurvedic and traditional physicians is mediated via antiglycation activity.

Absence of antimicrobial activity in alcoholic extract of Santalum album Linn

Vaishali Patil, GP Vadnere, Neeraj Patel

Journal of Pharmaceutical Negative Results 2011 2(2):107-109

Objective: The objective of the present study was to investigate the antimicrobial activity of seed of Santalum album Linn. (Family: Santalaceae) in various microbial strains. Materials and Methods: The effects of ethanolic extract of seed of S. album were analyzed in different microorganisms at a concentration of 5 mg/ml per disk. Results: The result indicated that the ethanolic extract of seed of S. album did not show any zone of inhibition against the tested microorganisms. Conclusion: The ethanolic extract of seed of S. album has no significant antimicrobial activity.

Anticonvulsant activity of Passiflora incarnata: No role of chrysin

Bhupinder Singh, Awanish Mishra, Rajesh Kumar Goel

Journal of Pharmaceutical Negative Results 2011 2(2):51-54

The earlier studies on Passiflora incarnata have pointed out the possible role of chrysin for its anticonvulsant activity. But role of chrysin in anticonvulsant property of P. incarnate seems to be controversial due to its poor bioavailability reported by different studies. Therefore, this study was designed to investigate the role of chrysin in anticonvulsant property of different extracts of P. incarnata used for medicinal purpose, viz. aqueous (PIAE), hydroethanolic (PIHE), and methanolic extracts (PIME). These extracts were prepared from dried leaves of P. incarnata using water, methanol, and 50% ethanol to obtain PIAE, PIME, and PIHE, respectively. The extracts were standardized with reference to chrysin by HPLC-UV method. Different doses (150, 300 and 600 mg/kg; i.p.) of PIAE, PIME, PIHE, and chrysin (1 mg/kg) were administered 30 min before the PTZ injection (75 mg/kg) to evaluate anticonvulsant effect. HPLC estimation has shown the higher amount of chrysin present in PIME followed by PIAE and PIHE. Significant dose-dependent delay in onset of convulsions was observed in PIHE and PIAE treated mice when compared with PTZ convulsive mice, while PIME treatment has not shown delay in onset of convulsions. Chrysin (1 mg/kg, i.p.), as well, did not produce significant increment in onset of convulsions. These results revealed that PIME containing significant amount of chrysin lacks anticonvulsant effect, while PIAE and PIHE, with insignificant chrysin content, have shown significant anticonvulsant effect. Thus, this study suggests that chrysin is not responsible for anticonvulsant effect of P. incarnata.

Rimonabant: Boom to ban

Suyog S Jain, Vitthal B Karande, Karuna B Ramteke, Girish T Raparti

Journal of Pharmaceutical Negative Results 2011 2(2):45-50

Rapidly rising prevalence of obesity and health care cost of its complications necessities need for highly efficacious and safe antiobesity drugs. As most old antiobesity drugs had moderate efficacy, severe toxicity and some of them very costly. Rimonabant, CB1 receptor antagonist was introduced with high expectations. Rimonabant has multiple beneficial effects, apart from significant weight loss it increases HDL and reduces triglycerides, Hb A 1C level, prevalence of metabolic syndrome. This wide spectrum of effects helps in comprehensive management of obesity and associated complications. Cost of rimonabant (generic Rs.5-8 per tablet) is also much lesser than the only other commonly used antiobesity drug orlistat around Rs. 40 per tablet. During trials rimonabant apart from nausea had neuropsychiatric side effects. EMEA approved rimonabant in June 2006 with concern over its psychiatric side effects. In 2007 EMEA contraindicated rimonabant in patients of depression or taking antidepressants. Further analysis in 2008 concluded that the incidence of psychiatric side effects was higher in clinical practice compared to controlled trials also few cases of suicide became a major safety concern, finally leading to suspension of its sale by EMEA in Oct 2008. USFDA never approved rimonabant over safety concerns, while India banned it shortly thereafter as precautionary measure. Present paper deals with the various events during the rapid rise and fall of rimonabant which was widely considered as blockbuster diet pill but was suspended over serious neuropsychiatric side effects.

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