Open Access Original Research Article

Fed Bioequivalence Studies for Immediate Release Drug Products: Beneficial or Wastage?

Suhas S. Khandave, Satish V. Sawant, Trupti V. Deshmukh, Farhat M. Shaikh, Urmila S. Khandave, Rakhi V. Sahane

Journal of Pharmaceutical Research International, Page 1696-1706
DOI: 10.9734/BJPR/2014/10496

Aims: The present study was undertaken to understand the need of fed bioequivalence studies for immediate release pharmaceutical products and thereby evaluating the relative appropriateness of guidelines given by the European Medicines Agency (EMA) and the United States Food and Drug Administration (USFDA). Vulnerability to show bioequivalent or non-bioequivalent results on the basis of type of drugs was also assessed.
Study Design: The present work is a meta-analysis involving 162 bioequivalence studies conducted on healthy human subjects.
Place and Duration of Study: Accutest Research Laboratories (I) Pvt Ltd, A-31, Khairne MIDC, TTC Industrial Area, Khairne, Navi Mumbai, 400 709, India, between June 2013 and February 2014.
Methodology: The present meta-analysis included a total of 162 bioequivalence studies of which 81 were conducted under fasted condition and the other 81 studies were conducted under fed condition. The drug products were fixed dose combinations and mono drug products for 22 and 140 studies respectively representing all the classes of Biopharmaceutics Classification System (BCS). The bioequivalence was assessed by standard criteria laid down by regulatory authorities. The results were correlated with the respective condition of study (fasted or fed) and the corresponding BCS class of the drug product. The observations were discussed in the light of available literature.
Results: A total of 78 and 74 studies conducted under fasting and fed conditions respectively had bioequivalent results. All studies conducted on fixed dose combination products had bioequivalent results. Five drug products had bioequivalent results only under fasting condition which proportionately contributed to 6.17% (5/81 fasting studies). These drug products complied only with the EMA guidelines and not USFDA defined passing criteria. All drug products belonging to BCS classes I and III showed bioequivalent results whereas drug products belonging to BCS class IV contributed to 80% of the total non-bioequivalent studies.
Conclusion: The EMA approach can be followed for BCS class I drugs while USFDA approach looks better for remaining drug products. Further research work is required to confirm the trend observed in our meta-analysis.

Open Access Original Research Article

HPTLC Methods for Determination of Gemifloxacin Mesylate in Rabbit Plasma

U. L. Narayan, B. Garnaik, S. K. Patro, S. Sahu

Journal of Pharmaceutical Research International, Page 1707-1714
DOI: 10.9734/BJPR/2014/4070

Aim: A new, simple, rapid, very sensitive and accurate high performance thin-layer chromatographic (HPTLC) method has been developed and validated for estimation of Gemifloxacin in rabbit plasma.
Study Design: Validation study.
Methodology: HPTLC was performed on silica gel 60F254 plates with ethanol: ethyl acetate: hexane, 2:7:1 (v/v), as mobile phase. Densitometry scanning was performed in absorbance mode at λ=254 nm.
Result: The RF value was 0.21. The response was a linear function of concentration over the range 0.1–0.7μg mL−1 (r2=0.996). A maximum recovery of drug from plasma was obtained by using chloroform and glacial acetic acid. Mean extraction recovery was 80%. Intra-day and inter-day precision (% RSD) of the assay were in the range 1.19–2.85% and accuracy was 1.7-5.66%
Conclusion: This method can be applied to pharmacokinetic studies in rabbit plasma.

Open Access Original Research Article

The Protective Role of Extract of Allium cepa Linn. (Liliaceae) (Red Onion) Bulb in Artesunate-induced Testicular Damage in Male Wistar Rats

Nwaehujor Chinaka O, Igile Godwin O, Nwinyi Florence C, Ode Julius O.

Journal of Pharmaceutical Research International, Page 1715-1724
DOI: 10.9734/BJPR/2014/10675

Aim: Allium cepa is consumed fresh or cooked in Nigeria and used as herb in ethno-medicinal practice against many ailments. The aqueous extract is used in the management of prostrate inflammation, alleviation of low sperm count and testicular damage related disorders in ethno medicine and folkloric medicine in the South-east regions of Nigeria. This study investigates the protective role of extract of Allium cepa bulb on spermatogenesis and testicular oxidative stress in male rats using Artesunate as testicular damage and oxidative stress inducing agent.
Study Design and Methodology: The design consisted of 5 groups of 10 male Wistar rats (140-180g) each. Groups 1, 2, 3 and 5 were given continuous oral challenge with 4.4mg/kgb.w Artesunate orally for two weeks to induce testicular damage. Animals were treated with 50, 150 and 300mg/kgb.w extract for 7 weeks. Groups 4 and 5 were not treated and served as normal and negative controls respectively. Animals were humanely euthanized, testes collected, homogenized and used for sperm count, motility and oxidative stress evaluation.
Results: Artesunate mediated testicular damage caused a significant (p<0.01) reduction in sperm count (17.5±2.4x106ml/l) compared to normal control (140X106ml/l). Extract treatment of groups 1, 2 and 3 caused a dose-dependent reversal of sperm count, motility and morphology after 7 weeks. Treatment with the highest dose (300 mg/kg) of the extract reversed testicular oxidative induced lesions as shown by levels of enzymatic antioxidants; CAT (10.112±1.73µg/mg), SOD (39.41±2.35µg/mg) and GPx (0.55±0.17nmol/mg) and non-enzymatic antioxidants; MDA (0.98±0.05nmol/mg) and GSH (2.17±0.05mg) relative to normal control, respectively.
Conclusion: These results demonstrated the potential beneficial effects of Allium cepa in the management of prostrate inflammation and testicular lesions.

Open Access Original Research Article

Antioxidant, Brine Shrimp Lethality and Antimicrobial Activities of Methanol and Ethyl-Acetate Extracts of Citrus macroptera Montr. Fruit Using In vitro Assay Models

Nizam Uddin, Md. Rakib Hasan, Md. Monir Hossain, Md. Mahadi Hasan, Aumit Roy, Tufikul Islam, Md. Sazzad Hossain, Abdullah Faruque, Md. Sohel Rana

Journal of Pharmaceutical Research International, Page 1725-1738
DOI: 10.9734/BJPR/2014/11235

Aims: To study and evaluate In vitro antioxidant, brine shrimp lethality and antimicrobial activities of both methanol and ethyl-acetate extracts of Citrus macroptera Montr. Fruit (Family-Rutaceae).
Study Design: In vitro antioxidant, brine shrimp lethality and antimicrobial activities.
Place and Duration of Study: Department of Pharmacy, Jahangirnagar University, Savar, Dhaka-1342. The study was carried out from November 2013 to January 2014.
Methodology: In vitro antioxidant activities of the extracts were studied using DPPH radical scavenging assay, NO scavenging assay, total phenol, total flavonoid content, total antioxidant capacity, total tannin content, lipid peroxidation by TBA, lipid peroxidation in human erythrocyte cell, reducing power capacity and cupric reducing capacity assays. Lethality bioassay was performed on Artemia salina Leach nauplii. Antimicrobial activity was investigated by disc diffusion technique.
Results: Methanol extract showed better activity than ethyl acetate extract in DPPH, NO, lipid peroxidation by TBA, reducing power capacity assay, total phenol, total flavonoid and total antioxidant capacity assays while ethyl –acetate extract showed more potency than methanol extract in total tannin content, cupric reducing capacity and lipid peroxidation in human erythrocyte assays. In brine shrimp bioassay both extracts showed promising lethal activity but methanol extract was found to be more potent than ethyl acetate extract (χ2=39.874, P<0.0001). In disc diffusion technique among six bacterial species, ethyl acetate extract showed broad spectrum antimicrobial activity against two gram positive Bacillus sublitis and Staphylococcus aureus and one gram negative Escherichia coli. 
Conclusion: The results demonstrate that methanol and ethyl-acetate extracts of C. macroptera fruit can be used as potential antioxidant, cytotoxic and antimicrobial agents. That is why extensive researches are necessary to search for active principles responsible for these activities.

Open Access Original Research Article

Activity Anti-Inflammatory and in silico Study of New Thiazolidinedione Derivatives

Laudelina R. de Magalhães, Iane Bezerra Vasconcelos Alves, Everaldo dos Santos, Vinícius Barros Ribeiro da Silva, Luiz Carlos Apolinário da Silva, Anekécia Lauro da Silva, Leidiane Carla Lira de Oliveira, Anne Cecília Nascimento da Cruz, Bruno Iraquitan Miranda da Silva, Lúcia Fernanda C. da Costa Leite, Marcelo Hernani Zaldini, Ivan da Rocha Pitta, Teresinha Gonçalves da Silva, Maria do Carmo Alves de Lima

Journal of Pharmaceutical Research International, Page 1739-1752
DOI: 10.9734/BJPR/2014/11294

Aims: Evaluation of the anti-inflammatory properties of new thiazolidine-2,4-diones derivatives.
Study Design: Study the effects of new thiazolidine-2,4-diones derivatives on the inflammatory process.
Place and Duration of Study: Departamento de Antibióticos, Universidade Federal de Pernambuco (UFPE), between June 2011 and July 2012.
Methodology: Compounds thiazolidine-2,4-diones were tested for anti-inflammatory activity by air pouch model. Swiss albino mice were used for the study. Air cavities were produced by subcutaneous injection of 2.5 mL of sterile air into the intrascapular area of the back. An additional 2.5 mL of air was injected into the cavity every 3 days to keep the space open. Seven days after the initial air injection, 1 mL of a 1% solution of carrageenan dissolved in saline was injected directly into the pouch to produce an inflammatory response. The compounds thiazolidine-2,4-diones and standard piroxicam were tested at doses of 3 mg/kg body weight. The total number of polymorphonuclear leukocytes (PMNL) was counted using an improved.
Results: The results support the use of these derivatives in inflammatory process. Among the compounds tested the ones that showed a greater effect in inhibiting the migration of neutrophils were the 3a, 3b, 3c, 3d and 3e. The anti-inflammatory effects showed by 3a-j were promising, probably due to the duality of action on PPAR alpha and gamma.
Conclusion: In conclusion, this study has shown that the thiazolidine derivatives do possess significant anti-inflammatory effects in laboratory animals. The exact mechanism and the bioactive principles responsible for these actions remain to be explained.

Open Access Original Research Article

Antibacterial and Antifungal Properties of Some Wild Nutraceutical Plant Species from Nebbi District, Uganda

Godwin Anywar, Hannington Oryem-Origa, Maud Kamatenesi-Mugisha

Journal of Pharmaceutical Research International, Page 1753-1761
DOI: 10.9734/BJPR/2014/11443

Aims: The purpose of this study was to investigate the antibacterial and antifungal properties of selected wild nutraceutical plants from Nebbi district in Uganda.
Study Design: Experimental study.
Place and Duration of Study: The study was carried out at the Department of Veterinary Microbiology and Parasitology, College of Veterinary Medicine, Animal Resources and BioSecurity, Makerere University, between January and March 2012.
Methodology: The diameters of the zones of inhibition and the Minimum Inhibitory Concentrations (MIC) were determined using the Agar well diffusion Assay and the serial dilution methods respectively.
Results: Seven plant species were tested for their antibacterial and antifungal activity against Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853) and Candida albicans (clinical isolates). The ether extract for Balanites aegyptiaca (L.) Delile showed the lowest MIC (150 µg/ml) against C. albicans, with a corresponding large diameter of the zone of inhibition (22.0 mm). The ether extract of Capparis erythrocarpos Isert. showed the highest activity against C. albicansand S. aureus, with MIC values of 330 µg/ml and 400 µg/ml respectively.
Conclusion: Balanites aegyptiaca (L.) Delile, and Capparis erythrocarpos Isert were the most potent antifungal and antibacterial nutraceutical plant species. The ether extract of Balanites aegyptiaca (L.) Delile, had the lowest MIC (150 µg/ml) against C. albicans and Capparis erythrocarpos Isert. showed the highest activity against C. albicans, S. aureus, and E. coli with MIC values of 330 µg/ml, 400 µg/ml and 320 µg/ml, respectively. The bioactivity of the selected wild nutriceutical plant species can be used to justify their ethnobotanical uses as ‘medicinal foods’.

Open Access Original Research Article

Use of Hydrazine Carboxamide-2-[(2-Hydroxy-1-Naphthalenyl) Methylene] as an Analytical Reagent for the Extractive Spectrophotometric Determination of U(Vi)

Ratnamala P. Sonawane

Journal of Pharmaceutical Research International, Page 1762-1771
DOI: 10.9734/BJPR/2014/7614

Extraction and spectrophotometric determination of Uranium (VI) was carried out using Hydrazine Carboxamide-2-[(2-Hydroxy-1-Naphthalenyl) Methylene] as an analytical reagent. A reddish brown coloured complex is formed at pH 9.2 between U(VI):HCHNM which is extracted in n-butanol. An intense peak for the complex was observed at 385nm, well separated from that of ligand. Beer’s law is obeyed over the range of 2 to20 ppm. Composition of M:L in the complex was ascertained by Job’s method, mole ratio method and slope ratio method and found to be 1:2. Extraction and spectrophotometric determination of U(VI) was carried out using Hydrazine Carboxamide-2-[(2-Hydroxy-1-Naphthalenyl) Methylene] as an analytical reagent. A reddish brown coloured complex is formed at pH 9.2 between U(VI):HCHNM which is extracted in n-butanol. An intence peak for the complex was observed at 385nm, well separated from that of ligand. Beer’s law is obeyed over the range of 2 to 20 ppm. Composition of M:L in the complex was ascertained by Job’s method, mole ratio method and slope ratio method and found to be 1:2. Diverse ion effect was studied for various ions and the proposed method was found to be highly selective for the trace concentration of U(VI). Molar absorptivity and Sandell sensitivity values calculated are 0.1566 x104L mol-1cm-1 and 0.0585μg/cmrespectively. Proposed method has been successfully applied to analyse synthetic samples and the results were found to be in good agreement with standard gravimetric methods.

Open Access Original Research Article

Evaluation of Ampicillin and Ceftriaxone on Ethanol Consumption as Compared to Naltrexone in Alcohol Dependant Rats

Kshitij Jadhav, Aafreen Saiyed, Suraj Ghag, Yashashri Shetty

Journal of Pharmaceutical Research International, Page 1772-1780
DOI: 10.9734/BJPR/2014/9473

Context: Extracellular glutamate level in reward centre of brain increases during ethanol drinking sessions. Hence, it can be hypothesized that drugs which decrease extracellular glutamate might have deaddictive properties. It has been shown that β-lactam antibiotics are potent stimulators of glutamate transporter 1(GLT1) expression. Previous studies have shown that ceftriaxone decreases ethanol consumption but this has not been compared to standard line of treatment (naltrexone). Also, no study was conducted for testing ampicillin even if in an in-vitro experiment ampicillin has shown to increase GLT1 levels more than ceftriaxone. Hence, our study’s objectives were to compare efficacy of ceftriaxone and ampicillin with naltrexone on ethanol consumption in rats.
Methods: Permission of ethics committee was taken. Study was divided into two parts. Part I included standardization of model & Part II included 8 groups of six rats each. Group 1: vehicle control, Group 2: 1mg/kg/d naltrexone, Group 3: 100mg/kg/d ceftriaxone, Group 4: 200mg/kg/d ceftriaxone, Group 5: 100mg/kg/d ampicillin, Group 6: 200mg/kg/d ampicillin were given i.p injections for 15 days and Group 7: 200mg/kg ceftriaxone & Group 8: 200mg/kg ampicillin i.p. single dose. Parameters measured were ethanol & water intake per day for 15 days.
Results: Groups 2 to 8 showed statistically significant decrease in ethanol intake as compared to vehicle control. Also, group 3 & 4 showed an increase in water consumption as compared to Group 1.
Conclusions: Our study recommends that drugs acting on glutamate pathways like ceftriaxone and ampicillin can be explored for treatment of alcohol dependence.

Open Access Original Research Article

Segregated Delivery of Rifampicin and Isoniazid from Fixed Dose Combination Bilayer Tablets for the Treatment of Tuberculosis

A. M. Silva, B. Abrahim-Vieira, F. A. do Carmo, L. H. do Amaral, L. C. Silva, C. S. Escudini, M. A. Lopes, V. P. Sousa, H. C. Castro, F. Veiga, C. R. Rodrigues, A. J. Ribeiro, L. M. Cabral

Journal of Pharmaceutical Research International, Page 1781-1801
DOI: 10.9734/BJPR/2014/11525

Aims: Develop an anti-tuberculosis (TB) Fixed Dose Combination (FDC) tablet containing an immediate release layer (IRL) composed of both rifampicin (RIF) and pyrazinamide (PYR) and a retarded release layer (RRL) comprised of isoniazid (INH) which would allow segregated delivery of RIF and INH.
Study Design: Trials were conducted on the pre-formulations and formulations to assess the compatibility of excipients and obtain a modified release profile, for an IRL consisting of both RIF and PYR and a RRL containing INH.
Place and Duration of Study: This study was performed at the Laboratory of Pharmaceutical Industrial Technology, Drug and Pharmaceutical Department, Faculty of Pharmacy, between March 2008 and April 2010.
Methodology: Preformulation studies were performed on RIF and PYR, alone and in combination with excipients. The pharmacopeic attributes of the distinct layers and the FDC tablets were evaluated for hardness, friability and disintegration time. The FDC bilayer tablets were analyzed for their drug content and cumulative dissolution of the drug.
Results: Fourier transform infrared, x-ray diffraction and differential scanning calorimetry results revealed the presence of amorphous and crystalline RIF forms and no potentially relevant incompatibilities were identified in the kneaded system containing RIF, PYR and excipients. In vitro drug release from the FDC tablets revealed that the intragranular addition of croscarmellose sodium into the IRL rendered a cumulative dissolution of RIF and PYR within the limits of simulated gastric fluid. And, for RRL, the most effective retardant matrix excipient was found to be hydroxypropyl methylcellulose.
Conclusion: Segregated delivery of RIF and INH from bilayer tablets is an option for the development of immediate release FDC tablets and the retarded release of INH, this strategy proved suitable for preventing contact of these two drugs under acidic conditions. This finding suggested that RIF had a high in vivo bioavailability which qualifies this FDC for future bioavailability studies.

Open Access Original Research Article

Tricyclic Pyrazoles: Synthesis, Characterization, Antimicrobial, Antituberculosis, and Antitumor Activity of N,1-Diphenyl-1,4-dihydrothiochro-meno[4,3-c]pyrazole-3-carboxamide-5,5-dioxide Derivatives

Pandaram Palanisamy, Sudalaiandi Kumaresan

Journal of Pharmaceutical Research International, Page 1802-1819
DOI: 10.9734/BJPR/2014/6155

Aims: A series of N,1-diphenyl-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxamide 5,5-dioxide derivatives (6a-m) were synthesized and evaluated for anticancer, antibacterial, and antifungal activity.
Methodology: Reaction of 2,3-dihydro-4H-thiochromen-4-one 1,1-dioxide 2 with diethyl oxalate in ethanol in the presence of a base afforded the Claisen condensation product 3. Subsequent reaction of 3 with phenylhydrazine hydrochloride at reflux in ethanol afforded ethyl 1-phenyl-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylate 5,5-dioxide (4). Alkaline hydrolysis of furnished the corresponding 1-phenyl-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylic acid 5,5-dioxide 5. The pyrazole acid 5 was converted into the corresponding acid chloride followed by treatment with an excess of the appropriate amine to give 6a-m.
Results: Compound 6k showed better activity than chloroamphenicol against Klebsiella pneumoniae and Escherichia coli and equipotent to clotrimazole in inhibiting the growth of Candida albicans (MIC 3.125 µg/mL). All compounds were screened for their cytotoxic activity against two tumor cell lines, namely, human colon tumor cell line (HCT116) and human cervical cancer cell line (HeLa) using the colorimetric MTT assay. Most of the tested compounds exhibited potent antitumor activity. Particularly, compound 6k displayed the highest activity among the tested compounds with IC50 equal to 17 µM (HeLa) and 15 µM (HCT116) respectively. Among the tested compounds, 6k was found to be more active against M. tuberculosis, (H37Rv) with minimum inhibitory concentration (7.8 µM).
Conclusion: The chloro- (6b and 6c), 2-aminobenzothiazole- (6l), and 4-aminoantipyrine- (6k) linkages exhibited better antimicrobial activity than their counterparts. Compound 6k was found to possess comparatively more antimicrobial, antituberculosis, and antitumor activity against the other derivatives.