DARU, Journal of Pharmaceutical Sciences
Volume:15 Issue: 3, Autumn 2007

Centaurea pamphylica Boiss. & Heldr. (Family: Asteraceae), commonly known as ‘pamphylia daisy'', is a Turkish endemic species of the genus Centaurea that comprises ca. 500 species, many of which have been used as traditional medicines. The n-hexane, dichloromethane (DCM) and methanol (MeOH) extracts of the aerial parts of C. pamphylica were assessed for antioxidant activity and general toxicity using, respectively, the 2,2-diphenyl-1-picrylhydrazyl (DPPH), and the brine shrimp lethality assays. The reversed-phase preparative HPLC and PTLC were used to isolate compounds from the extracts. The structures of these compounds [1-4] were elucidated by spectroscopic means, and also by direct comparison with the respective published data. Both the DCM and the MeOH extract showed significant levels of antioxidant activities with the RC50 values of 72.6 x 10-2 and 47.3 ´ 10-2 mg/mL, respectively. The MeOH extract exhibited low levels of toxicity towards brine shrimps (LD50 = 125.0 ´ 10-2 mg/mL). Three major bioactive components of the MeOH extract were matairesinoside [1], arctiin [2] and matairesinol [3]. An eudesmane-type sesquiterpene, pterodontriol [4], was also isolated from the DCM extract. Since reactive oxygen species are important contributors to various ailments, the antioxidant properties of the extracts as well as the isolated compounds may be of medicinal significance. This is the first report on the occurrence of 1-4 in C. pamphylica, and 4 in the genus Centaurea.

Tecoma stans (L.) Juss or Yellow bells from Bignoniaceae is a ornamental tropical shrub or small tree predominantly found in central, and south America and in Latin America is used traditionally for reducing blood glucose. However, its other pharmacological effects have not been yet elucidated. The aim of present study was to investigate the effect of its leaves extract on rat ileum contractility and involved mechanism(s). Tecoma stans Juss hydroalcoholic leaf extract (TLE) was prepared by macerated method using 70% alcohol. Distal segment of ileum (2 cm) from male Wistar rat was mounted in an organ bath containing Tyrode solution (10 ml, pH 7, 37 °C) and pre-contracted by carbachol (CCh, 10 µM) or by KCl (60 mM). The antispasmodic effects of TLE (0.125-2 mg/ml) were studied prior and after 20-30 min incubation of ileum with propranolol (1µM), naloxone (1µM), L-NAME (100 µM), or 5 min incubation with glibenclamide (10 µM) and tetraethylammonium (TEA, 1mM). The effect of TLE on CaCl2-induced contraction in Ca2+-free with high K+ Tyrode solution was also studied. The CCh- and KCl-induced ileal contractions were reduced by TLE (P<0.0001). This effect was not attenuated by propranolol, naloxone, L-NAME, glibenclamide and TEA. In Ca2+-free Tyrode solution with high K+, cumulative concentrations of CaCl2 induced contractions which were inhibited by TLE dose-dependently. Our results indicate that the Tecoma stans (L.) Juss leaf extract induces its antispasmodic effects without involvement β-adrenoceptors, opioid receptors, potassium channels and NO production. It seems that, the calcium channels are involved in this spasmolytic effect.

Chimonanthus fragrans Lindle (Calycanthaceae) is an aromatic plant which little information has been reported so far on the composition of its essential oil. In this study the essential oil of flower of this plant was obtained by hydrodistillation and analyzed by GC and GC-MS. Forty nine components were identified corresponding to ca. 98.12 % of the total components of the essential oil with 0.12 % yield. The major components were elemol (20.06%), caryophyllene (9.51%), elemene (8.65%), bicyclogermacrene (8.15%), elemene (7.2%), germacrene-D (5.65%), transocimene (5.5%), sabinene (3.65%), linalool (2.6%), caryophyllene oxide (2.3%), and cadinene (1.95%). Comparison of the data of this study with other data including recent report by HS-SPME-GC-MS showed quantitative and qualitative differences due to geographical, agricultural, and technical factors.

A sensitive, accurate and rapid reverse phase HPLC method was developed to quantitate plasma levels of diclofenac sodium in human plasma. The drug, internal standard (naproxene) and orthophosphoric acid 1 M, were added to plasma samples and vortexed for 20 sec. A mixture of hexane /isopropyl alcohol (90:10) was then added and vortexed for 2 min. Samples were centrifuged and the supernatant layer was separated, evaporated to dryness under nitrogen gas stream, reconstituted in mobile phase and an aliquot of 50 μl was analyzed on a μ-bondapack C18 (150 × 4.6mm) column, with 45% acetonitrile in deionised water and 0.5% orthophosphoric acid, (pH = 3.5) at 276 nm. The standard curve covering 0.005 - 4 μg/ml concentration range, was linear, relative errors were within 0.13 to 16 % and the CV % ranged from 1.24 to 8.75. The limits of quantitation and detection of the method were 0.005 μg/mL and 0.002μg/mL, respectively. The method was suitable for bioavailability and pharmacokinetic studies of diclofenac in humans and applied in a randomized, two-way cross over bioequivalance study of two different diclofenac sodium preparations with twelve subjects and with a one-week washout perio

Ocular inserts of ofloxacin were prepared with objectives of reducing the frequency of administration, obtaining controlled release and greater therapeutic efficacy in the treatment of eye infections such as conjunctivitis, keratitis, corneal ulcers, etc. Polyvinyl alcohol (PVA) ofloxacin films were prepared by mercury substrate method. The ocular inserts were evaluated for drug-excipient interaction, physico-chemical characteristics, microbiological and in vitro and in vivo release studies. There was no interaction between drug and excipients as revealed by UV absorption and IR spectra of the pure drug, medicated and placebo formulations. The weight and thickness of the inserts were in the range of 57.3-126.0 mg and 55.6-99.3 microns, respectively for different formulations; casting of rate-controlling membrane increased the weight and thickness of the inserts. Tensile strength and percent elongation at break varied with the nature of rate-controlling membrane and film thickness. Moisture vapour transmission through films followed zero-order kinetics and decreased with increase in film thickness. The drug content varied from 99.53-99.86%. The method of exposure to UV radiation was used for sterilization of ocular inserts and no microbial growth was observed in any formulation during sterility testing by direct inoculation method. The influence of rate-controlling membrane of different polymers of ethyl cellulose (EC) and polymethacrylates (Eudragit RL100, Eudragit RS100) on release kinetics was studied. The drug release for prepared formulations with rate-controlling membrane of EC, Eudragit RS100 (ERS), Eudragit RL 100 (ERL) was found to be 85.80, 93.85 and 98.71%, respectively and followed zero-order kinetics. Ocular insert F3 with rate-controlling membrane of Eudragit RS100, when inserted into the eye of rabbit showed controlled release up to 24 hours. There was a good correlation between in vitro and in vivo release data. The developed formulation was effective against selected microorganism during in vitro antimicrobial efficacy studies. PVA-ofloxacin ocular inserts with rate-controlling membrane of Eudragit RS100 are promising for controlled ocular delivery of ofloxacin.

Isosteviol has been found to have potential preventive or therapeutic effects against hypertension, ischemia reperfusion injury, diabetes and cancer, but little is known about the pharmacokinetics (PK) of the compound. The aim of this study was to develop a liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method for determination of isosteviol in rat plasma and to assess in a preliminary manner the PK of isosteviol after intravenous bolus injection.Ions of analytes were generated using electro-spray ionization and detected in the positive-ion mode in LC-MS/MS. Multiple reaction monitoring was performed, using the precursor product ion combination for isosteviol m/z 319.4→273.4. Progesterone was used as an internal standard. Nitrogen was used as the nebulising gas and unit resolution was set for Q1 and Q3. Isosteviol solution was injected through the penile vein of rats at a dose of 8 mg/kg. Blood samples were collected from a jugular vein cannula. The PK parameters were calculated using a two - compartment PK model.The LC-MS/MS assay for isosteviol in rat plasma was linear over the range of 0.5-80 μg/ml. The terminal half life of isosteviol (t 1/2) was 406 ± 31.7 min and clearance (CL) was 2.9 ± 0.3 ml/min/kg. A sensitive LC-MS/MS assay for isosteviol in plasma has been successfully established and used in a preliminary PK evaluation of isosteviol in rats.

Immunotoxins, which are composed of both the cell targeting and the cell killing moieties are the new approach for targeted therapy of human disease. In all immunotoxins that GM-CSF has been used as cell targeting; only cell lines expressing high affinity receptor have been used for cytotoxicity studies. In the present study, various cell lines expressing high and low affinity receptors were used for assessment of the cytotoxic effect of hybrid chimeric protein. The expression of GM-CSF receptor (GM-CSFR) was quantified by real-time RT- PCR. The cell lines K562 and THP1 expressing high affinity receptor and MC-7, PC-3 and DU145 expressing low affinity receptor were used for this study. The chimeric hybrid protein was found to be toxic for various cell lines used in this investigation and cytotoxicity was more effective in cell lines bearing high affinity receptors. Overall, our results showed that the recombinant hybrid protein could have wide range of application on various cancer cell lines even cells bearing low affinity receptors for GM-CSF.

The effect of morphine on spatial learning and memory is controversial. In the present study, the male rats were used to evaluate the effect of morphine dependence and cold water swimming on spatial learning and memory. Dependent animals received morphine sulfate in drinking water for 25 days. Animals were divided into four groups in simple randomized manner. The first control and dependent groups, were studied in normal water (20± 2 ◦C), and the second control and dependent groups were studied in cold water (10-12◦ C). Morris Water Maze (MWM) experimentations were begun from 21st to 25th days of morphine administration. In each group of animals, spatial learning and memory parameters were analyzed. The results showed that morphine dependence may facilitates spatial learning and memory in MWM, and cold water reduces swimming speed but facilitating the formation of spatial memory. On the other hand, cold water swimming abolished the effects of morphine on spatial learning but facilitated its effect on spatial memory. The underling mechanism(s) to these phenomenon remains to be elucidated.

The methanol extract of the leaves of Ficus carica Linn. (Moraceae) was evaluated for hepatoprotective activity in rats with liver damage induced by carbon tetrachloride. The extract at an oral dose of 500 mg/kg exhibited a significant protective effect by lowering the serum levels of aspartate aminotransferase, alanine aminotransferase, total serum bilirubin, and malondialdehyde equivalent, an index of lipid peroxidation of the liver. These biochemical observations were supplemented by histopathological examination of liver sections. The activity of extract was also comparable to that of silymarin, a known hepatoprotective.

The diverse biological effects of hepatocyte growth factor/scatter factor (HGF/SF) are mediated by c-Met which is preferentially expressed on epithelial cells. Met signaling has a role in normal cellular activities, and may be associated with development and progression of malignant processes. In this study presence of Met in the axillary drainage from patients who underwent conservative operations for breast cancer, and its prognostic significance was examined. Sixty-two consecutive patients with invasive ductal carcinoma of the breast which were suitable for breast-conserving treatment participated in the study. The output of the drain that had been placed in the axilla during the operation was collected, and the presence of Met and β-actin were assessed by reverse transcriptase-polymerase chain reaction (RT-PCR) assays. The data were compared with the pathological features of the tumor and the axillary lymph nodes, and with the estrogen and progesterone receptors status.RT-PCR of the axillary lymphatic drainage was positive for Met in 46 (74.2%) of the patients and positive assays were correlated with increase in tumor size and grade of capillary and lymphatic invasion, as well as with lymph node metastasis (P < 0.02, for all comparisons). All 24 patients with axillary lymph node metastases in comparison with those without lymph node (57.9%) metastases had positive assays for Met. While all ten patients with tumor involvement in the margins of the resection had positive assays for Met in their lymphatic fluid, only 36 out of 52 patients (69.2%) were positive for met assay. Finally, Met showed negative correlations with positive estrogen and progesterone receptor assays (P<0.02).From the results of this study it may concluded that Met can be detected in the axillary fluids of patients with breast cancer and its expression in the axillary drainage may be a potential prognostic factor. This finding might be useful in therapeutic considerations since a positive assay for Met in histologically node-negative patients might indicates the need to search for node microinvasion or involvement of the excision margins with tumor