In vitro and in vivo immunomodulatory activities of iridoids fraction from Barleria prionitis Linn.

ETHNOPHARMACOLOGICAL RELEVANCE: Barleria prionitis Linn. (Family: Acanthaceae), one of the important Ayurvedic medicinal plant in India, has long been used to treat variety of ailments including swellings, gout, arthritic and rheumatic disorders, nervine and skin diseases, and also acts as immunorestorative. AIM OF THE STUDY: The present study was aimed to explore in vitro and in vivo immunomodulatory activities of the iridoids fraction i.e. n-butanol fraction of methanol extract from Barleria prionitis aerial parts (IFBp). MATERIALS AND METHODS: IFBp was studied for in vitro [nitroblue tetrazolium (NBT) test and neutrophils candidacidal assay] and in vivo immunomodulatory activity on cellular and humoral immune responses to the antigenic challenge by sheep red blood cells (SRBCs) and by neutrophil adhesion test, phagocytic activity and cyclophosphamide-induced myelosuppression. The study comprised the preliminary phytochemical screening, HPTLC standardization and maximum tolerable dose determination of IFBp. RESULTS: IFBp (50, 100 and 200µg/ml) significantly (P≤0.01) increased the intracellular killing activity of stimulated neutrophils assayed by in vitro NBT reduction test and neutrophils candidacidal assay. Pretreatment of IFBp (100 and 200mg/kg; p.o.) evoked a significant increase in percent neutrophils and neutrophils adhesion to nylon fibres. Oral administration of IFBp augmented the humoral immune response to SRBCs, evidenced by increase in antibody titres and dose dependently potentiated the delayed-type hypersensitivity reaction induced by SRBCs in mice. IFBp potentiated significantly (P≤0.01) the macrophage phagocytic activity and ameliorated the red blood cells, total white blood cells and platelets count and haemoglobin concentration, and also restored the myelosuppressive effects induced by cyclophosphamide. The content (% w/w; mean±SD, n=3) of main iridoids i.e. shanzhiside methyl ester and barlerin was found to be 21.55±2.40 and 10.03±1.69 in IFBp of BP, respectively. CONCLUSION: The present investigation reveals that IFBp is a potent immunostimulant, stimulating both the specific and non-specific immune mechanisms.

Solid State Characterization of Domperidone: Hydroxypropyl-ß-Cyclodextrin Inclusion Complex.

The purpose of the present study was to prepare inclusion complex of domperidone with hydroxylpropyl-ß-cyclodextrin in order improved the solubility and hence to increase dissolution of domperidone. An effect of concentration of hydroxylpropyl-ß-cyclodextrin on the aqueous solubility of domperidone was determined by phase-solubility method. The aqueous solubility of domperidone increased as a function of hydroxylpropyl-ß-cyclodextrin concentration, showing AL type diagram. Solid domperidone/hydroxylpropyl-ß-cyclodextrin complex was prepared in ratio 1:1 by ultrasonication and kneading method. Solid state inclusion complex was characterized by FTIR, powder X-ray diffraction and differential-scanning calorimetry techniques. FTIR studies showed intactness of drug in complex whereas powder diffraction studies showed that hydroxylpropyl-ß-cyclodextrin complex was amorphous. Solubility studies showed that complexation increased domperidone solubility as compared to pure drug in 0.1M hydrochloric acid and distilled water. Drug content confirms that ultrasonication is one of the efficient methods to prepare inclusion complex. Dissolution data of inclusion complexes also indicated that there is 1.4 folds increase in dissolution as compared to pure drug and was observed in case of inclusion complexes prepared by ultrasonication.

Simultaneous Determination of Ofloxacin and Ornidazole in Solid Dosage Form by RP-HPLC and HPTLC Techniques.

The objective of this work was to develop and validate simple, rapid and accurate chromatographic methods for simultaneous determination of ofloxacin and ornidazole in solid dosage form. The first method was based on reversed phase high performance liquid chromatography, on Intersil C(18) column (250 mm, 4.6 i.d.), using acetonitrile:methanol: 0.025M phosphate buffer, pH 3.0 (30:10:60 % v/v/v) as the mobile phase, at a flow rate of 1 ml/min at ambient temperature. Quantification was achieved with UV detection at 318 nm over a concentration range of 2-40 µg/ml for ofloxacin and 5-100 µg/ml for ornidazole. The mean retention time of ofloxacin and ornidazole was found to be 4.04 min and 5.83 min, 6.77 min (isomers), respectively. The amount of ofloxacin and ornidazole estimated as percentage of label claimed was found to be 100.23 and 99.61%, with mean percent recoveries 100.20 and 100.93%, respectively. The second method was based on TLC separation of these drugs using silica gel 60F(254) aluminium sheets and dichloromethane:methanol:25% ammonia solution (9.5:1:3 drops v/v) as mobile phase. Detection was carried out at 318 nm over the concentration range of 20-100 ng/spot for ofloxacin and 50-250 ng/spot for ornidazole. The mean R(f) value of ofloxacin and ornidazole was found to be 0.16 and 0.56, 0.78 (isomers), respectively. The amount of ofloxacin and ornidazole estimated as percentage of label claimed was found to be 100.23 and 99.61% with mean percent recoveries 100.47 and 99.32%, respectively. Both these methods were found to be simple, precise, accurate, selective and rapid and could be successfully applied for the determination of pure laboratory prepared mixtures and tablets.

Antihyperlipidemic effect of the methanolic extract from Lagenaria siceraria Stand. fruit in hyperlipidemic rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Lagenaria siceraria Stand. (Cucurbitaceae) fruits have been used, traditionally, in the treatment of hyperlipidemia and atherosclerotic impasse and considered as cardioprotective and cardiotonic drug. AIM OF THE STUDY: The present work was undertaken to investigate the effects of methanolic extract from Lagenaria siceraria in experimentally induced hyperlipidemia in rats. MATERIALS AND METHODS: Methanolic extract of Lagenaria siceraria fruits (LSFE) (100, 200 and 300 mg/kg; p.o.) was administered to the high fat-diet-induced hyperlipidemic rats for 30 days to evaluate its antihyperlipidemic activity. Atorvastatin (10mg/kg; p.o.) was used as a standard drug. RESULTS: At the 30 th day, most significant reduction in lipid levels in the LSFE treated rats as compared to the rats fed with high-fat diet at the 0th day were: total cholesterol 290.14+/-18.42 mg/dl vs. 228.58+/-16.38 mg/dl, low-density lipoprotein cholesterol 195.14+/-8.86 mg/dl vs. 120.57+/-8.11 mg/dl, triglyceride 232.41+/-15.22 mg/dl vs. 181.79+/-15.68 mg/dl, very low-density lipoprotein cholesterol 46.48+/-3.04 mg/dl vs. 36.35+/-3.13 mg/dl (P<0.0001). Conversely, high-density lipoprotein cholesterol levels were significantly (P<0.0001) increased from 48.52+/-6.52 to 71.66+/-5.14 mg/dl. The increase in weight in rats administered with LSFE was less when compared to rats fed with high-fat diet. Moreover, LSFE also exhibited significant increase in excretion of bile acids. CONCLUSION: The results demonstrate that the LSFE has a definite antihyperlipidemic potential. There is also a valid scientific basis for consuming it in the treatment of coronary artery diseases in India.

Effect of atorvastatin and hydroxychloroquine combination on blood glucose in alloxan-induced diabetic rats.

OBJECTIVE: To evaluate the antihyperglycemic activity of atorvastatin and hydroxychloroquine combination in alloxan-induced diabetic rats. MATERIALS AND METHODS: Alloxan induced diabetic Wistar male rats were randomized into six groups of 6 rats each. (Normal rats, diabetic control, atorvastatin (ATV), hydroxychloroquine (HCQ), ATV 5 mg /kg + HCQ 100 mg/kg, and ATV 10 mg/kg + HCQ 200 mg/kg). The rats were treated for 9 days and blood samples were collected at baseline and end of therapy. These samples were analyzed for plasma glucose by autoanalyzer. Changes in body weight, water, food intakes and total protein content were also recorded. RESULTS: Atorvastatin and hydroxychloroquine alone and in combination reported significant fall in blood glucose level from baseline. Fall in glucose level was significantly more in high dose combination of atorvastatin and hydroxychloroquine (ATV: 10 mg/kg + HCQ: 200 mg/kg) as compared to other study treatment groups (ATV: 17% Vs HCQ: 7% Vs ATV 5mg/kg + HCQ 100mg /kg: 14% Vs ATV 10mg/kg + HCQ 200mg /kg: 21%; p<0.01). ATV and HCQ individually and in combination also improved the body weight loss. The weight gain was significantly more in combination treated rats as compared to positive control group and greater than those who received atorvastatin and hydroxychloroquine alone. Rats treated with the combination also reported significant decrease in food intake and significant increase in total protein. CONCLUSION: Increased hypoglycemic effect in combination may be due to potentiation or synergism between HCQ and ATV. Further studies are required to demonstrate clinically significant antidiabetic effect.

Intranasal mucoadhesive buspirone formulation: in vitro characterization and nasal clearance studies.

Oral administration is unsuitable for drugs prone to extensive first-pass metabolism, like buspirone. Thus, in the present study an attempt has been made to develop a mucoadhesive intranasal formulation improving permeation characteristics of buspirone HCl. Nasal formulations containing different concentrations of chitosan HCl and hydroxypropyl-beta-cyclodextrins (HP-beta-CD) were prepared and compared with control buspirone HCl solution regarding permeability, in vitro duration of mucoadhesion, in vivo nasal clearance in rats and in vitro cytotoxicity on cell culture. Nearly two fold increase in buspirone permeation was observed with 1% chitosan HCl and a 3.5 fold increase with 1% chitosan HCI and 5% HP-beta-CD. Nasal clearance studies showed retention of 50% radioactivity up to about 3.5 h for formulation F7 containing 1% chitosan HCI compared to 1.5 h for control buspirone solution (F1). Results conclusively demonstrated enhancement in permeation with no cytotoxicity. Thus formulations can be used to improve bioavailability of buspirone HCl.

Simultaneous spectrophotometric determination of lansoprazole and domperidone in capsule dosage form.

Two simple, accurate and precise spectrophotometric methods have been developed for simultaneous determination of lansoprazole and domperidone in pharmaceutical dosage form. Method A involves formation of Q-absorbance equation at 256.0 nm (isoabsorptive point) and at 294.2 nm while method B is two wavelength method where 277.6 nm, 302.1 nm were selected as lambda(1) and lambda(2) for determination of lansoprazole and 231.3 nm, 292.0 nm were selected as lambda(1) and lambda(2) for determination of domperidone. Both the methods were validated statistically and recovery studies were carried out. The Beer's law limits for each drug individually and in mixture was within the concentration range of 5-50 mug/ml. Linearity of lansoprazole and domperidone were in the range of 24-36 mug/ml and 8-12 mug/ml, respectively. The proposed methods have been applied successfully to the analysis of the cited drugs either in pure form or in pharmaceutical formulations with good accuracy and precision. The method herein described can be employed for quality control and routine analysis of drugs in pharmaceutical formulations.

Development and in vitro evaluation of buccoadhesive tablets of metoprolol tartrate.

Buccoadhesive tablet of metoprolol tartrate was developed to prolong its release and improve bioavailability by avoidance of hepatic first pass metabolism during the treatment of chronic hypertension. The formulations were tested for weight, hardness, friability, content uniformity, swelling index, bioadhesive force and drug release rate. Carbopol 934 P was used as bioadhesive polymer and methocel K4M was added as a matrix former. Backing layer of ethyl cellulose was given to the tablets. Optimised formulation containing carbopol 934 P and methocel K4M in the ratio of 1:1 showed surface pH values in the range of 6 to 7 and 91.50% cumulative release of drug in 10 h. Stability study revealed that the optimized formulation was stable for atleast 3 mo.

A Validated HPTLC Method for Determination of Ondansetron in Combination with Omeprazole or Rabeprazole in Solid Dosage Form.

A simple, precise, accurate and rapid high performance thin layer chromatographic method has been developed for the simultaneous estimation of ondansetron combinations in solid dosage form with omeprazole and rabeprazole, respectively. The method involved separation of components by TLC on a precoated silica gel 60 F(254) using a mixture of dichloromethane:methanol (9:1) as a mobile phase. Detection of spots was carried out at 309 nm and 294 nm for ondansetron with omeprazole and ondansetron with rabeprazole combinations, respectively. The mean retardation factor for ondansetron and omeprazole were found to be 0.42+/-0.02, 0.54+/-0.03, respectively while for ondansetron and rabeprazole, 0.41+/- 0.02 and 0.51+/-0.02, respectively. The linearity and range was 0.1 to 0.5 mug/spot for three drugs. The method was validated for precision, accuracy and reproducibility.

Simultaneous spectrophotometric estimation of paracetamol and metoclopramide hydrochloride in solid dosage form.

Two simple, accurate, rapid and economical spectrophotometric methods have been developed for the simultaneous determination of paracetamol and metoclopramide hydrochloride from tablet dosage form. The first method developed employs formation and solving simultaneous equations using 248.6 nm and 275.6 nm as two wavelengths for formation of equations. Second method is absorbance ratio in which wavelengths selected were 265.6 nm as isoabsorptive point and 275.6 nm as lambdamax of paracetamol. Both the drugs and their mixtures obey Beer-Lamberts law at selected wavelengths at given concentration range. The methods have been validated statistically and by recovery studies. The results of analysis have been validated statistically and by recovery studies.

Analgesic and antipyretic effects of Capparis zeylanica leaves.

The ethanol and water extracts of Capparis zeylanica leaves showed dose-dependent and significant (P<0.05) increases in pain threshold in tail-immersion test. Moreover, both the extracts (100-200 mg/kg) exhibited a dose-dependent inhibition of writhing and also showed a significant (P<0.001) inhibition of both phases of the formalin pain test. The water extract (200 mg/kg) significantly (P<0.01) reversed yeast-induced fever. Preliminary phytochemical screening of the extracts showed the presence of alkaloids, flavonoids, saponins glycosides, terpenoids, tannins, proteins and carbohydrates.

Immunostimulant effects of Capparis zeylanica Linn. leaves.

The present study was undertaken to explore the immunomodulatory activity of ethanolic and water extracts of Capparis zeylanica Linn. (family: Capparidaceae) leaves on neutrophil adhesion test, humoral response to sheep red blood cells, delayed-type hypersensitivity, phagocytic activity and cyclophosphamide-induced myelosuppression. Pre-treatment of water extract (300 mg/kg, oral) of Capparis zeylanica evoked a significant increase in neutrophil adhesion to nylon fibres. The augmentation of humoral immune response to sheep red blood cells by ethanolic and water extracts (150-300 mg/kg) is evidenced by increase in antibody titres in mice. A dose-related increase in both primary and secondary antibody titre was observed. Oral administration of ethanolic and water extracts of Capparis zeylanica leaves, at doses of 150 and 300 mg/kg in mice, dose dependently potentiated the delayed-type hypersensitivity reaction induced by sheep red blood cells. Immunomodulatory activity was also assessed by serological and haematological tests. Capparis zeylanica extracts prevented myelosuppression in mice treated with cyclophosphamide drug. The study comprised the acute toxicity and preliminary phytochemical screening of the ethanol and water extracts.

Hypolipidemic and antihyperlipidemic effects of Lagenaria siceraria (Mol.) fruit extracts.

Bottle gourd [(Lagenaria siceraria (Mol.) Stand.] fruit is ascribed with many therapeutic effects. The present study was undertaken to explore the antihyperlipidemic effect of four different extracts viz. petroleum ether, chloroform, alcoholic and aqueous extracts from bottle gourd in Triton-induced hyperlipidemic rats and their hypolipidemic effects in normocholesteremic rats. The study is comprised preliminary phytochemical screening of the extracts. Oral administration of the extracts, at doses of 200 and 400 mg/kg body weight in rats, dose-dependently inhibited the total cholesterol, triglycerides, low-density lipoproteins level, and significantly increased the high density lipoproteins level. However, petroleum ether extract did not show the significant effects. Both the chloroform and alcoholic extract exhibited more significant effects in lowering total cholesterol, triglycerides and low density lipoproteins along with increase in HDL as compared to the others. Preliminary phytochemical screening revealed the presence of flavonoids, sterols, cucurbitacin saponins, polyphenolics, proteins, and carbohydrates. The results obtained suggest marked antihyperlipidemic and hypolipidemic activity of the extracts.