Appraisal of Transdermal Water-in-Oil Nanoemulgel of Selegiline HCl for the Effective Management of Parkinson's Disease: Pharmacodynamic, Pharmacokinetic, and Biochemical Investigations.

In the present study, the potential of transdermal nanoemulsion gel of selegiline hydrochloride for the treatment of Parkinson's disease was investigated. Water-in-oil nanoemulsions were developed by comparing low- and high-energy methods and were subjected to thermodynamic stability tests, in vitro permeation, and characterization studies. In vitro studies indicated that components of nanoemulsion acted as permeation enhancers with highest flux of 3.531 ± 1.94 µg/cm2/h from nanoemulsion SB6 containing 0.5 mg selegiline hydrochloride, 3% distilled water, 21% S mix (Span 85, Tween 80, PEG 400), and 76% isopropyl myristate by weight. SB6 with the least droplet size of 183.4 ± 0.35 nm, polydispersity index of 0.42 ± 0.06 with pH of 5.9 ± 0.32 and viscosity of 22.42 ± 0.14 cps was converted to nanoemulsion gel NEGS4 (viscosity = 22,200 ± 400 cps) by addition of Viscup160® for ease of application and evaluated for permeation, safety, and pharmacokinetic profile in Wistar rats. It provided enhancement ratio 3.69 times greater than conventional gel. NEGS4 showed 6.56 and 5.53 times increase in bioavailability in comparison to tablet and conventional gel, respectively, along with sustained effect. Therefore, the developed water-in-oil nanoemulsion gel promises to be an effective vehicle for transdermal delivery of selegiline hydrochloride.

3D-QSAR study of 8-azabicyclo[3.2.1] octane analogs antagonists of cholinergic receptor.

3D-QSAR models of Comparative of Molecular Field Analysis (CoMFA) and Comparative of Molecular Similarities Indices Analysis (CoMSIA) of 20 8-azabicyclo[3.2.1] octane (potent muscarinic receptor blocker) was performed. These benztropine analogs were optimized using ligand based alignment method. The conventional ligand-based 3D-QSAR studies were performed based on the low energy conformations employing database alignment rule. The ligand-based model gave q(2) value 0.819 and 0.810 and r(2) value 0.991 and 0.988 for CoMFA and CoMSIA, respectively, and the predictive ability of the model was validated. Results indicate that the CoMFA and CoMSIA models could be reliable model which may be used in the design of novel muscarinic antagonists as leads.

Evaluation of dosage forms. VI. Studies of commercial oxyphenbutazone tablet dosage forms.

Dissolution-dialysis studies of commercial tablets of oxyphenbutazone were carried out to establish the applicability of this technique for the in vitro evaluation of oxyphenbutazone dosage form. While disintegration time and dissolution rate studies did not give a true indication of bioavailability, an excellent correlation was obtained between the dialysis rate constant K and the pharmacokinetic parameters AUC and Cmax.

Evaluation of dosage forms. IV. Studies on commercial phenylbutazone tablet dosage forms.

In order to determine the feasibility of dissolution-dialysis as a suitable technique for in vitro evaluation, studies on commercial phenylbutazone tablets were carried out. Although disintegration time and dissolution parameters did not give a true indication of bioavailability, an excellent correlation was obtained between the dialysis rate constant (K) and various pharmacokinetic parameters obtained from bioavailability studies on human volunteers.

Modification of the glycine (co-activator) binding site of the N-methyl-D-aspartate receptor in the guinea pig fetus brain during development following hypoxia.

The present study was designed to investigate the mechanism of NMDA receptor activation as a function of brain maturation by studying the development of the glycine binding site of the NMDA receptor and its modification by in-utero hypoxia in the guinea pig fetus brain during gestation. Measurements of Bmax (number of functional receptors) and Kd (apparent receptor affinity) of glycine binding sites of the NMDA receptor were performed in eleven (45 days, n = 5; 60 days, n = 6) synaptosomal membranes from normoxic (control) fetuses and ten (45 days, n = 4; 60 days, n = 6) synaptosomal membranes constituted the hypoxic (experimental) group. In the experimental group, fetuses were exposed to maternal hypoxia (FiO2 0.07) for 1 h. Synaptosomal membranes were prepared and strychnine-insensitive specific [3H]glycine binding was determined During development, the number of glycine binding sites increased (Bmax:392 +/- 30 vs. 583 +/- 30 fmol/mg protein at 45 and 60 days respectively, P < 0.05) where as the affinity remained unchanged (Kd: 190 +/- 9 vs. 211 +/- 30 nM at 45 and 60 days respectively). Following hypoxia, glycine binding sites increased at 45 days (Bmax:392 +/- 30 vs. 561 +/- 96 fmol/mg protein, P < 0.005) but decreased at 60 days (Bmax:583 +/- 85 vs. 411 +/- 65 fmol/mg protein, P < 0.005) with change in Kd only at 60 days (Kd:211 +/- 30 vs. 149 +/- 52 nM, P < 0.05). The data show that there are alterations in the characteristics of the glycine binding site during development and following hypoxia. We conclude that developmental changes in the glycine binding site might modulate NMDA receptor activation as a function of brain maturation. Furthermore, hypoxia-induced modification of the glycine binding site might be a potential mechanism of neurotoxicity and might increase susceptibility of the fetal brain to excitotoxicity at term.

Protective effect of alpha-tocopherol on brain cell membrane function during cerebral cortical hypoxia in newborn piglets.

Protective effect of alpha-tocopherol on the structure and function of brain cell membranes was investigated by measuring Na+,K(+)-ATPase activity and products of lipid peroxidation (fluorescent compounds) in brain cell membranes obtained from newborn piglets. Four groups of anesthetized, ventilated piglets were studied: five hypoxic piglets and five normoxic piglets were pretreated with free alpha-tocopherol (20 mg/kg/dose i.m.), five additional hypoxic piglets received i.m. placebo and five normoxic piglets served as control. Placebo and alpha-tocopherol were given 48 and 3 h prior to onset of hypoxia. Hypoxic hypoxia was induced and cerebral hypoxia was documented as a decrease in the ratio of phosphocreatine to inorganic phosphate (PCr/P(i)) using 31P NMR spectroscopy. PCr/P(i) decreased from baseline of 2.62 +/- 0.54 to 1.05 +/- 0.27 in alpha-tocopherol-pretreated and from 2.44 +/- 0.48 to 1.14 +/- 0.30 in the placebo-pretreated group during hypoxia. Na+,K(+)-ATPase activity was unchanged in both normoxic and hypoxic alpha-tocopherol-pretreated groups. However, in placebo-pretreated hypoxic group, Na+,K(+)-ATPase activity decreased as compared with control (44.9 +/- 9.7 vs. 61.8 +/- 5.7 mumol P(i)/mg protein/h, P < 0.005). The level of fluorescent compounds increased in placebo-pretreated but not in alpha-tocopherol-pretreated group as compared with control. During hypoxia, serum alpha-tocopherol levels were higher in alpha-tocopherol-pretreated groups as compared with placebo-pretreated hypoxic group. The present data indicates that alpha-tocopherol protects brain cell membranes in newborn piglets from lipid peroxidative damage during tissue hypoxia probably by being incorporated in cell membrane and also as circulating antioxidant.

Effect of allopurinol on uric acid levels and brain cell membrane Na+,K(+)-ATPase activity during hypoxia in newborn piglets.

Oxygen-free radicals generated by xanthine oxidase during hypoxia-ischemia may result in cellular injury through harmful effects on membrane phospholipids. The present study investigated the effect of administration of allopurinol, an inhibitor of xanthine oxidase, on free-radical generation and brain cell membrane injury during hypoxia by inhibiting the breakdown of hypoxanthine to uric acid. Brain cell membrane Na+,K(+)-ATPase activity and lipid peroxidation products (conjugated dienes and fluorescent compounds) were determined as indices of brain membrane function and structure. Cerebral oxygenation was continuously monitored during hypoxia by 31P-NMR spectroscopy. Plasma and brain tissue levels of uric acid were measured to evaluate xanthine oxidase activity and purine degradation. Na+,K(+)-ATPase activity decreased significantly in both hypoxic groups; however, the allopurinol-treated hypoxic group showed a smaller decrease than the untreated hypoxic group (47.3 +/- 4.9 vs. 42.0 +/- 2.7 mumol Pi/mg protein/h, P < 0.05), respectively. Conjugated dienes increased significantly in the untreated hypoxic compared to control animals (0.070 +/- 0.045 vs. 0.004 +/- 0.006 mumol/g brain, P < 0.05), with the allopurinol-treated animals having intermediate values (0.053 +/- 0.039 mumol/g brain). Fluorescent compounds were lower in the allopurinol-treated hypoxic group compared to the untreated hypoxic group (0.79 +/- 0.19 vs. 1.06 +/- 0.60 micrograms/quinine sulfate/g brain, P < 0.05). Measurements of serum and brain tissue uric acid were significantly lower during hypoxia in the allopurinol-treated compared to the untreated group (30.3 +/- 15.6 vs. 45.7 +/- 10.6 microM (P < 0.05) and 1.69 +/- 0.97 vs. 4.27 +/- 2.37 nmol/g (P < 0.05), respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Modification of NMDA receptor by in vitro lipid peroxidation in fetal guinea pig brain.

The effect of lipid peroxidation on the NMDA receptor and its modulatory sites in fetal guinea pig brain cell membranes was examined. P2 membrane fractions were prepared from the fetal brain tissue and peroxidized in the presence of ferric chloride and ascorbate. [3H]-MK-801-binding studies were performed and Bmax (number of binding sites) and Kd (affinity) values were used as indices of NMDA receptor modification. In lipid-peroxidized membranes the Kd value increased from 6.76 +/- 2.69 in control to 15.12 +/- 7.38 nM (P < 0.01), indicating a decreased affinity of NMDA receptors following lipid peroxidation. However, there was no significant change in Bmax. The glutamate- and glycine-dependent increase in activation was 40% lower in lipid-peroxidized membranes as compared to control. The spermine-dependent activation was also significantly reduced following lipid peroxidation as compared to control suggesting decreased affinity of spermine site. The results of this study indicate that lipid peroxidation modifies recognition, coactivator and spermine sites of NMDA receptor by decreasing its affinity without affecting the number of binding sites. Normal activation of NMDA receptor is important for neuritic growth, synaptogenesis, long-term potentiation and synaptic plasticity. Therefore, we speculate that any clinical condition causing lipid peroxidation of brain cell membranes could jeopardize these maturational processes in the developing brain causing neurological impairment.

Selective sensitivity of synaptosomal membrane function to cerebral cortical hypoxia in newborn piglets.

The effect of hypoxia on the structure and function of the synaptosomal membranes and myelin fraction (glial cells, neuronal cells bodies and axonal membranes) was investigated by measuring Na+,K(+)-ATPase activity and levels of lipid peroxidation products in cerebral cortical synaptosomal membranes and myelin fractions obtained from newborn piglets. Hypoxic hypoxia was induced and cerebral hypoxia was documented as a decrease in the ratio of phosphocreatine to inorganic phosphate (PCr/Pi) using 31P-NMR spectroscopy. PCr/Pi decreased from baseline of 2.93 +/- 0.76 to 0.61 +/- 0.36 during hypoxia. The synaptosomal membrane Na+,K(+)-ATPase activity decreased from a control value of 56.6 +/- 3.7 to 40.4 +/- 6.0 mumol Pi/mg protein/h during hypoxia. The level of conjugated dienes increased from zero (reference value) to 4.5 +/- 2.7 nmol/mg lipid and the level of fluorescent compounds increased from 23.5 +/- 2.2 to 92.6 +/- 46.4 ng quinine sulfate/mg lipid in the synaptosomal membranes during hypoxia. No change in myelin fraction Na+,K(+)-ATPase activity or levels of lipid peroxidation products were noted. These data indicate that synaptosomal membranes, rich in polyunsaturated fatty acids, are more susceptible to oxygen free radical mediated lipid peroxidative damage during hypoxia.