DARU, Journal of Pharmaceutical Sciences
Volume:18 Issue: 1, Spring 2010

Background and the purpose of the study: Lamotrigine (LMG) undergoes extensive hepatic metabolism upon oral administration and its absorption is affected in the presence of food. This study was aimed to develop nanosuspension of LMG and investigate its formulation characteristics using L9 orthogonal array. Nanosuspension was prepared using emulsification-solvent diffusion method. All the formulations were subjected to in-vitro evaluation and the statistically optimized one was used for stability, scanning electron microscopic and differential scanning calorimetric studies. Nanoparticles were spherical with little surface adsorbed drug. Formulation characteristics in terms of size, zeta potential, polydispersity index (PDI), entrapment efficiency (EE), drug content and in vitro drug release were consistent and within their acceptable range. All the batches provided a burst release profile during first 1 hr, followed by a controlled release extending up to 24 hrs. The values of n in Peppas model ranged between 0.2-0.4 for all the formulations indicative of Fickian release mechanism. The formulation remained reasonably stable up to 3 months. No interaction was observed among the drug and polymers. Major Results of in vitro drug release studies suggested that nanosuspension might be used as a sustained delivery vehicle for LMG. Statistical analysis revealed that size of the nanoparticles was most strongly affected by stabilizer type while EE was influenced by the drug-to-polymer ratio.

Background and the purpose of the study: Lercanidipine hydrochloride (LRDP) is used in the treatment of hypertension because of its selectivity and specificity on the smooth vascular cells. The pharmacokinetic parameters make LRDP a suitable candidate for transdermal delivery. The purpose of the study was to select a suitable formulation for the development of transdermal drug-delivery system (TDDS) of LRDP and to determine the effect of penetration enhancer, limonene on drug permeation. The matrix type TDDS of LRDP were prepared by solvent evaporation technique. Formulations A1, A2, A3, A4, A5 and A6 were composed of Eudragit RL100 (ERL) and hydroxypropyl methyl cellulose (HPMC) in 1.5:8.5, 3:7, 4:6, 6:4, 7:3 and 8.5:1.5 ratios respectively. All the six formulations carried 10 mg of LRDP/patch area, 8 % v/w of d-limonene as a penetration enhancer, 20 % v/w of propylene glycol as plasticizer in methanol and dichloromethane as solvent system. The prepared TDDS were evaluated for physicochemical characteristics, in-vitro release, ex-vivo permeation and skin irritation. The ex-vivo permeation studies were carried out across excised rat skin using Franz diffusion cell. All the formulations exhibited satisfactory physicochemical characteristics. Cumulative percentage of the drug released in 24 hrs from the six formulations were 82.0 %, 74.9 %, 63.2 %, 63.5 %, 59.8 % and 53.5 % respectively. Corresponding values for the cumulative amounts of the drug permeated across the rat skin for the above matrix films were 2644.5, 2347.2, 2249.5, 1933.4, 2021.5 and 1663.4 µg/cm2 respectively. By fitting the data into zero order, first order and Higuchi model, it was concluded that drug release from matrix films followed Higuchi model and the mechanism of the drug release was diffusion mediated. The patches were seemingly free of potentially hazardous skin irritation. The patches composed of ERL, HPMC (1.5:8.5) with 8 % v/w limonene as penetration enhancer may be selected for the development of TDDS of LRDP for potential therapeutic use by using a suitable adhesive layer and backing membrane.

Background and the purpose of the study: MEN1 is an important tumor suppressor gene that encodes a nuclear protein called menin. Recent data suggest that interactions between menin and other proteins have important roles in control of the cell cycle and apoptosis. In addition، estrogen receptor (ER)، an important prognostic factor is differentially expressed in breast cancer cells. In this study the MEN1 gene and protein expression in MCF7، T47D and MDA-MB-468 breast cancer cell lines with different ER status following exposure to adriamycin (ADR) was investigated. Cytotoxicity of ADR on these cell lines was determined using MTT assay. The mRNA and protein levels were analyzed in tested cell lines using RT-PCR and immunocytochemistry (ICC) assays، respectively. ADR cytotoxicity was highest on MDA-MB-468 and lowest on MCF7 cells. MEN1 mRNA showed significant decrease after ADR exposure only in the MDA-MB-468 cell line. Menin protein expression was higher in MDA-MB-468 and lower in MCF7 cells. Differential molecular responses to adriamycin were observed in cancer cell lines. Molecular data also suggest that MEN1 as a new biomarker can be used in combination with current biomarkers for prediction of response to chemotherapy.

Background and the purpose of the study: Analysis of current immunomodulating strategies indicates that monovalent approaches are unlikely to restore immunostasis or achieve complete therapy of sepsis. Setarud (IMOD) as a mixture of urtica، carotenoids، urea، and selenium has been recently patented for its potential in reduction of Tumor Necrosis Factor alpha (TNF-α) and Interferon-γ and Interleukin-2 levels. The aim of this study was to examine efficacy of IMOD in the management of patients with severe sepsis. Twenty patients with severe sepsis and acute physiology and chronic health evaluation (APACHE) score of more than 20 were randomized to receive standard treatment of severe sepsis (control group) or standard treatment plus IMOD (IMOD group). The group treated with IMOD for 14 days was according to the pilot study and regarding the stability of patient''s conditions in the ICU. Of course patients in both groups received standard treatment and all were monitored for 28 days. Blood samples were analyzed for interleukins (IL-1، IL-2، IL-6)، plasminogen activator inhibitor (PAI-1)، TNF-α، total thiol molecules (TTM)، nitric oxide (NO)، total antioxidant power (TAP)، and lipid peroxidation (LPO). Daily APACHE، Sequential Organ Failure Assessment (SOFA)، and Simplified Acute Physiology Score (SAPS) were calculated. Results and major Comparing with controls، IMOD was significantly effective in improving SAPS، SOFA، and APACHE scores، and reduction of mortality rate. Among tested inflammatory biomarkers، IMOD significantly improved TTM and TNF-α values. It is concluded that IMOD might be added as a safe adjutant to standard treatment of severe sepsis.

Background and the purpose of the study: The hippocampal formation is involved in nociception. Prenatal serotonin depletion results in a significant decrease in the concentration of nociceptive sensitivity during the second phase of behavioral response in the formalin test. A microdialysis probe was inserted via a guide cannula into the right CA1 region of the hippocampus. Extracellular serotonin (5HT) and its 5- hydroxyindoleacetic acid (5HIAA) metabolite overflow were collected every 10 min during the formalin test and measured by HPLC with electrochemichal detector. Compared to the sham group، formalin injection in the hind paw of the rat significantly increased 5HT after 10، 30، 40، and 50 min and increased 5HIAA after 10، 30، 40، 50، and 60 min collection time periods in hippocampal dialysate. (n=6 for each group at each sampling time). In the formalin treated rats serotonin and 5HIAA concentrations increased in the biphasic pattern in concert with the first and second phases of formalin pain. The hippocampal formation might be involved in the processing of nociceptive information and serotonin-related mechanisms in the hippocampus may play a role in the biphasic behavioral responses to formalin noxious stimulation.

Background and the Purpose of the study: Central Angiotensin Converting Enzyme (ACE) has an important role on cerebral microcirculation and metabolism. However، its role in terms of protecting the brain from ischemic/reperfusion (I/R) injury are debatable. This study evaluated the role of ACE، using enalapril as ACE inhibitor، in protection of the brain from I/R injury during transient focal cerebral ischemia (TFCI) in normotensive rat. Male Sprague Dawley rats (280-320g) randomly assigned to control ischemic and enalapril pre-treated ischemic groups. Enalapril was injected intraperitoneally 1 h before middle cerebral artery occlusion (MCAO) at the dose of 0. 03 or 0. 1 mg/kg. Cerebral ischemia was induced by 60 min MCAO followed by 24 hrs reperfusion. After evaluation of neurological deficit scores (NDS) the animal was sacrificed for assessment of cerebral infarction and edema. TFCI induced cerebral infarctions (283±18 mm3)، brain edema (4. 1±0. 4%) and swelling (9. 8±1. 5%) with NDS of 3. 11±0. 36. Non-hypotensive dose of enalapril (0. 03 mg/kg) improved NDS (1. 37±0. 26)، reduced cerebral infarction (45%)، brain edema (54%) and swelling of the lesioned hemispheres (34%) significantly. However، hypotensive dose of enalapril (0. 1 mg/kg) could improve neurological activity (1. 67±0. 31) and failed to reduce cerebral infarction (276±39mm3) and swelling (10. 4±1. 4%). In the rat model of transient focal cerebral ischemia، inhibition of angiotensin converting enzyme with non-hypotensive doses of enalapril has the benefit of improving neurological activity، reducing cerebral infarction، brain swelling and edema of acute ischemic stroke. Therefore، it is reasonable to conclude that central renin-angiotensin system may participate in ischemic/reperfusion injury of the cerebral cortex.

Background and the purpose of the study: The available literatures show that 5-HT1A receptors are widely distributed throughout the basal ganglia، and their activation facilitate dopamine release. Neuroleptic drugs such as haloperidol induce Parkinson-like syndrome through blocking brain D2 receptors. This study aimed to investigate effect of buspirone، a partial agonist of 5HT1A receptor، on motor dysfunctions induced by haloperidol and involvement of 5HT1A receptors in this regard. Study was performed on the male mice weighing 25-30 g. Animals were divided randomly to groups of 10 animals. Motor dysfunction was induced by intraperitoneal (i. p.) injection of haloperidol (1 mg/kg). Catalepsy was assayed by bar-test method 5، 60، 120 and 180 minutes after drug administration and motor imbalance was studied by rotarod test. Results and major Results showed that buspirone (20 mg/kg، i. p.) decreased significantly haloperidol-induced catalepsy and balance disorder in a dose dependent manner. Furthermore، 8-OH-DPAT (10 mg/kg، i. p.)، as an agonist of 5-HT1A receptor، decreased haloperidol-induced catalepsy and balance disorder. The effect of buspirone (20 mg/kg، i. p.) on haloperidol-induced motor disorders was abolished by NAN-190 (10 mg/kg، i. p.)، as a 5-HT1A receptor antagonist. From the results it may be concluded that buspirone improves haloperidol-induced catalepsy and balance disorder through activation of 5-HT1A receptors.

Background and the Purpose of the Study: Oral mucositis is one of the most common complications of malignancy chemotherapy. As yet، no absolute treatment has been demonstrated to be effective for chemotherapy- induced oral mucositis. This study evaluates the effectiveness of phenytoin mouthwash as a wound healing agent، on the basis of stimulating effects on fibroblast proliferation. In this multicenter، randomized، placebo- controlled clinical trial; twelve patients received phenytoin mouthwash (0. 5%) or placebo for about two weeks. Oral pain severity was scored on the daily basis using a VAS (visual analogue scale) of 10 centimeters. National Cancer Institute (NCI) scale was used to grade the intensity of mucositis. To determine the effect of treatment، a quality of life questionnaire، consisting of 35 queries، was filled out for all patients. Statistical analyses of data was performed using Mann- Whitney test. The average time for complete remission of mucositis in phenytoin- treated group was less than that of the placebo group. The quality of life improved dramatically in the phenytoin group with the healing process being more evident in the first week. Furthermore، reduction in the wound area was greater in the phenytoin group than controls at the end of the first week of treatment. Both groups eventually demonstrated reduction in pain intensity; however no statistically significant difference was observed between two groups. Phenytoin mouthwash accelerated wound healing and resolution of mucositis and improved life quality impressively.

Backgrounds and the purpose of the study: Inducible NO synthase activity has been frequently reported in varicose veins. Aminoguanidine is known to inhibit iNOS. The aim of this study was to examine the effects of aminoguanidine on varicocelized rats. Male Wistar rats were divided into groups A، B، C، D، E، and F (control group). Groups A، B، C، and D rats underwent left varicocele induction with a 20-gauge needle. Group E (sham) rats underwent a similar procedure، but the renal vein was left intact. Ten weeks after varicocele induction، sperm parameters were evaluated in groups D، E، and F. Groups A and B received 50 mg/kg aminoguanidine or placebo، respectively، daily for 10 weeks. After 10 and 20 weeks of varicocele induction، the fertility outcomes of the experimental groups were evaluated. The values of the sperm parameters did not differ significantly between groups B and D، but were significant when compared with groups F and E (P ≤ 0. 05). The values of the sperm parameters of groups F and E showed no significant changes (P ≤ 0. 05). The changes between group A and groups B and D were significant (P ≤ 0. 05). Ten weeks after varicocele induction، rats of groups A، B، and C were still fertile. After 20 weeks، only half of the rats in group A were fertile. Aminoguanidine improved the sperm parameters and mating outcomes in vari-cocelized rats.

Background and the purpose of the study: Bifidobacterial strains are excessively sensitive to acidic conditions and this can affect their living ability in the stomach and fermented foods، and as a result، restrict their use as live probiotic cultures. The aim of the present study was to obtain bifidobacterial isolates with augmented tolerance to simulated gastrointestinal condition using cross-protection method. Individual bifidobacterial strains were treated in acidic environment and also in media containing bile salts and NaCl. Viability of the acid and acid-bile-NaCl tolerant isolates was further examined in simulated gastric and small intestine by subsequent incubation of the probiotic bacteria in the corresponding media for 120 min. Antipathogenic activities of the adapted isolates were compared with those of the original strains. Results and major The acid and acid-bile-NaCl adapted isolates showed improved viabilities significantly (p<0. 05) in simulated gastric fluid compared to their parent strains. The levels of reduction in bacterial count (Log cfu/ml) of the acid and acid-bile-NaCl adapted isolates obtained in simulated gastric fluid ranged from 0. 64-3. 06 and 0. 36-2. 43 logarithmic units after 120 min of incubation. There was no significant difference between the viability of the acid-bile-NaCl-tolerant isolates and the original strains in simulated small intestinal condition except for Bifidobacterium adolescentis (p<0. 05). The presence of 15 ml of supernatants of acid-bile-NaCl-adapted isolates and also those of the initial Bifidobacterium strains inhibited pathogenic bacterial growth for 24 hrs. Probiotic bacteria with improved ability to survive in harsh gastrointestinal environment could be obtained by subsequent treatment of the strains in acid، bile salts and NaCl environments.

Background and the purpose of the study: Heat Shock Protein 90 (Hsp90) is typically the most abundant chaperone in the eukaryotic cell cytoplasm، and its expression is essential for loading immunogenic peptides onto major histocompatibility complex molecules for presentation to T-cells. Therefore، it may act as a good candidate as an adjuvant molecule in vaccine technology. Initially the human Hsp90β gene was cloned into the heat inducible expression vector pGP1-2 and then the recombinant protein was isolated by ion exchange chromatography. After intradermal injection of confirmed purified band of protein to rabbits and isolation of the serum IgG antibody، for its affinity purification، the rabbit’s purified Hsp90 specific IgG was coupled to the cyanogen bromide-activated Sepharose 4B. The recovery of the purified protein of interest by affinity chromatography was 50%. This research enabled purification of human heat shock protein by a laboratoryprepared column chromatography

Background and the purpose of the study: The species Hymenocrater calycinus، belongs to the plant family Lamiaceae and grows wildly in the north-east of Iran. Previously، the antimicrobial activity of the plant extracts was reported. In the present study، the bioactivity-guided fractionation of the methanol extract of H. calycinus and the combination effects of the isolated compound with cell wall active agents against S. aureus and E. coli was investigated. Column and thin layer chromatographic methods were used for isolation and purification and spectroscopic data (MS، 1H- and 13C-NMR، HMQC، HMBC and 1H-1H COSY) were employed for identification of the compound isolated from the extract. A disk diffusion method was used to determine the antibacterial activity of the isolated compound against S. aureus and E. coli in comparison with 7 different antibiotics. The isolated compound 1 was identified as 3- (3، 4- dihydroxyphenyl) lactic acid 2-O-quinic acid. Compound 1 (500 µg/disc) enhanced antibacterial effect of ampicillin، ciprofloxacin، vancomycin and cefepime against S. aureus and activated the effects of ampicillin and vancomycin against E. coli. Results showed that the compound 1 was not active against both tested strains at any concentration below 1 mg/disk، and as a result the enhancing effect of the compound could be due its association with antibiotics.