QUALITY BY DESIGN-BASED OPTIMIZATION AND VALIDATION OF NEW REVERSE PHASE-HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR SIMULTANEOUS ESTIMATION OF LEVOFLOXACIN HEMIHYDRATE AND AMBROXOL HYDROCHLORIDE IN BULK AND ITS PHARMACEUTICAL DOSAGE FORM

ABSTRACTObjective: Innovative application of quality by design (QbD) technique for simultaneous estimation of levofloxacin and ambroxol hydrochloride (HCL)in bulk and its pharmaceutical dosage form using reverse phase-high-performance liquid chromatography (RP-HPLC) method.Method: A method has been developed for the separation of levofloxacin and ambroxol HCL using RP-HPLC on C18 column (250 4.6 mm, 5 ml) withultraviolet detection at 306 nm. Experimental designs were applied for multivariate optimization of the experimental conditions of RP-HPLC method.Three independent factors: Acetonitrile content in the mobile phase composition, buffer pH, and flow rate were used to design mathematical models.Here, central composite design (CCD) experimental design was used to study the response surface technique and to study in depth the effects ofthese independent factors. Derringer's desirability function was applied to simultaneously optimize the retention time of last eluting peak (ambroxolhydrochloride) and resolution between levofloxacin and ambroxol hydrochloride.Result and Discussion: The predicted optimum assay condition consisted of acetonitrile, potassium dihydrogen phosphate buffer (pH 5.00;potassium dihydrogen phosphate), and methanol in a proportion of 20:70:10% v/v, respectively, as the mobile phase at a flow rate of 1.2 ml/minute.Using this optimum condition, baseline separation of both drugs with good resolution and a run time of <5 minutes were achieved. The optimizedassay condition was validated according to the ICH guidelines to confirm specificity, linearity, accuracy, and precision.Keywords: Levofloxacin, Ambroxol hydrochloride, Experimental design, Response surface methodology, Derringer's desirability, Quality by designapproach

A STUDY ON PATIENTS AWARENESS ON RATIONAL USE OF ANTIBIOTICS AND ITS RESISTANCE

Objectives: Antibiotics are needed for many serious illnesses such as bacterial pneumonia, bacterial meningitis, septicemia, and even strep throat.These illnesses can be life threatening or can lead to serious complications. The work aims to create an awareness on antibiotics and its resistance inpatients. The main objective of this work is to study the patient knowledge through knowledge assessment questionnaire, to promote the rational useof antibiotics and to educate the patients using antibiotics.Methods: A questionnaire was prepared and data were collected from the patients based on which the study was carried out.Results and Conclusion: A very high consumption of antibiotics was observed.there was a higher use of cephalosporins. Dispensing of antibioticsis very high in community pharmacy despite of federal regulations, heath education programs should be taken to the patients regarding antibiotics.Keywords: Rational use, Antibiotics, Resistance

SYNTHETIC ROUTES AND NATURAL SOURCES OF 2-PYRIDONE DERIVATIVES AND THEIR PHARMACOLOGICAL ACTIVITY

Objective: 2-pyridone is a well-known heterocyclic ring having significant biological and medical application. The molecular structures and various activities of 2-pyridone derivatives as well as their syntheses and natural occurrence are analyzed and reviewed, and their reactivity toward various nucleophiles is discussed. Methods: 2-pyridone derivatives, first naturally obtained and described as early as before the 19th century, have been attracting increasing attention in view of their high reactivity as building blocks for the preparation of compounds of various classes due to their selective transformations with different reagents. Much information describing the natural occurrence, synthesis and the significant biological activity of 2-pyridone compounds are scattered throughout the literature. There are short chapters dealing with the synthesis and activity of 2-pyridone derivatives. Results: After compiling the above material, the abundance of certain heterocyclic ring and nature of typical chemical transformations applied in current drug synthesis. It is likely this results from the abundance of these heterocycles in natural products such as alkaloids and various synthetic derivatives revealing different biological activity. This might suggest a classical approach to drug design where substrate analogs gain inspiration from existing natural ligands. Conclusions: The data considered in this review clearly demonstrate the high synthetic potential of 2-pyridone derivatives. Many biologically active heterocyclic compounds have been obtained based on this heterocyclic ring. This suggests that 2-pyridone can be used in the design of novel highly effective pharmaceuticals with a broad spectrum of bioresponses

PREPARATION AND CHARACTERIZATION OF GEMCITABINE LOADED MPEG-PCL POLYMERIC NANOPARTICLES FOR IMPROVED TRANSPORTATION ACROSS BLOOD BRAIN BARRIER

Objective: To prepare Gemcitabine (GCB) loaded Methoxy Polyethylene Glycol-Poly (Caprolactone), (MPEG-PCL) nanoformulations and to carry out the physicochemical characterisation with a primary objective to enhance the transport and penetration of drug across the blood-brain barrier (BBB).Methods: Gemcitabine loaded MPEG-PCL nanoparticles were formulated by using modified nanoprecipitation method. Nanoformulations were prepared by varying drug: polymer ratio. The prepared nanoparticles (NP) were evaluated for particle size, zeta potential, entrapment efficiency, drug content and in-vitro drug release studies. The in vitro cytotoxicity of drug-loaded NPs was evaluated in U-87 MG cells. Results: The prepared nanoformulations indicated a significant increase in particle size with increase in the polymeric concentration. GCB loaded MPEG-PCL nanoformulation (GCBNP 3) exhibited a particle size of 223±1.4 nm. DSC thermo grams indicated that GCB was dispersed as an amorphous state in MPEG NPs. SEM, TEM, and AFM studies indicated that the NPs were spherical, smooth surface without any cracks or pinholes. In vitro studies showed the GCBNP 3 shows an initial burst release followed by a more gradual and sustained-release phase (maximum drug release). The cytotoxicity of GCB loaded MPEG-PCL nanoformulations showed reduction in the IC50 value (4.1 µg). Apoptosis detection assay with Hoechst 33342 dye was carried out and observed an increase in fluorescence in the apoptotic cells. By invasive studies, the GCB loaded MPEG-PCL nanoformulation inhibits the cell migration significantly when compared with the pure drug.Conclusion: The GCB loaded MPEG-PCL nano particles indicated improved cytotoxic activity with minimal concentrations compared with the pure drug in U-87 MG glial cells. Hence, it can be concluded that GCB loaded MPEG-PCL nanoformulation can serve as a potential drug delivery tool for the treatment of brain tumours.Â

Inhibitory effects of flavonoids isolated from Givotia rottleriformis bark and Cassia tora leaves on the production of pro-inflammatory cytokines in LPS stimulated human whole blood

The plants Givotia rottleriformis bark and Cassia tora leaves were used in indigenous medicine in the treatment of chronic inflammatory diseases like psoriasis. In previous work, three flavonoids namely Rutin (I), Luteolin-7-O-β-D-Glucuronide (II) and Kaempferol 3-O-[2-O-(6-O-feruloyl)-β-D-glucopyranosyl]-β-D-galactopyranoside (III) were isolated from G. rottleriformis bark and three flavonoids viz quercetin-3-O-β-d-glucuronide (IV), Luteolin-7-O-β-glucopyranoside (V) and Formononetin-7-O-β-D-Glucoside (VI) were isolated from C. tora leaves and evaluated for antipsoriatic activity. The cytotoxic effect of isolated compounds I-VI was evaluated using HaCaT cells, a rapidly multiplying human keratinocyte cell line, as a model of epidermal hyperproliferation in psoriasis. Among the isolated compounds, compound II, III and VI showed significant antiproliferant activity in HaCaT cells. Pro-inflammatory cytokines viz., IL-1α, IL-1β, IL-6, IL-8, IL-17 and TNF-α contributes to the pathogenesis of chronic inflammatory skin diseases such as psoriasis and has been a target for the development of the new anti-psoriatic drug. Hence the present study aimed to evaluate inhibitory effects of ethanol extract of selected plants and isolated compound II, III and VI (5-40 mg/mL) on lipopolysaccharide (LPS) induced pro-inflammatory cytokines viz., IL-1α, IL-1β, IL-6, IL-8, IL-17 and TNF-α production. The level of IL-1α, IL-1β, IL-6, IL-8, IL-17 and TNF-α pro-inflammatory cytokines were detected using enzyme-linked immunosorbent assay. The ethanol extract of both the plants was standardized by HPLC using chemical markers. Preincubation with isolated compounds II, III, VI and ethanol extract of selected plants strongly attenuated LPS-induced increase in the concentrations of IL-6 (50-73 %) and TNF-α (64-90 %). The compound II and III also showed significant inhibition (*p ≤0.05) of IL-1β, IL-8 and IL-17. The results showed that the selected plants and isolated flavonoids have potential effects as anti-inflammatory agents by inhibiting the release of pro-inflammatory cytokines supporting the folkloric utilization

Evaluation of herbal ointment containing ethanol extract of Plecranthus amboinicus root for the management of psoriasis

Plant, Plecranthus amboinicus (Lour.) Spreng, belonging to the family Lamiaceae, commonly known as ‘Karpuravalli’ in Tamil language is widely used in folk medicine to treat conditions like cold, asthma, constipation, headache, cough, fever and skin diseases. The present study aimed to evaluate antipsoriatic effect of the ethanol extract of Plecranthus amboinicus root. Ointment containing ethanolic extract of Plecranthus amboinicus root was prepared and evaluated for antipsoriatic activity using complete Freund's adjuvant (CFA) and formaldehyde induced animal model. Psoriasis is induced by applying mixture of 0.1 mL of prepared CFA and formaldehyde mixture (1:10 ratio) topically for 7 days on the dorsum surface of the skin of Swiss albino mice. Antipsoriatic effect of 0.5% and 1.0% (w/w) ointments containing ethanolic extract of Plecranthus amboinicus root was evaluated in terms of Psoriasis severity index (PSI) by the phenotypic features (redness, scales and erythema) and histological features (epidermal thickness). The result showed that there was a significant increase in the orthokeratinocyte layer and a significant reduction in the epidermal layer of skin in the in vivo mice model with a progressive reduction (p**<0.01) in the severity of psoriatic lesions (redness, erythema, and scales) from day 7 to 21st day and significant (p*<0.05) decreased epidermal thickness and increased orthokeratotic regions in animals treated with 0.5% and 1.0% (w/w) ointments of Plecranthus amboinicus root. The present investigations revealed that Plecranthus amboinicus root possess potent antipsoriatic activity, confirming their traditional use in skin disorders

Evaluation of herbal ointment containing ethanol extract of Plecranthus amboinicus root for the management of psoriasis

553-559Plant, Plecranthus amboinicus (Lour.) Spreng, belonging to the family Lamiaceae, commonly known as ‘Karpuravalli’ in Tamil language is widely used in folk medicine to treat conditions like cold, asthma, constipation, headache, cough, fever and skin diseases. The present study aimed to evaluate antipsoriatic effect of the ethanol extract of Plecranthus amboinicus root. Ointment containing ethanolic extract of Plecranthus amboinicus root was prepared and evaluated for antipsoriatic activity using complete Freund's adjuvant (CFA) and formaldehyde induced animal model. Psoriasis is induced by applying mixture of 0.1 mL of prepared CFA and formaldehyde mixture (1:10 ratio) topically for 7 days on the dorsum surface of the skin of Swiss albino mice. Antipsoriatic effect of 0.5% and 1.0% (w/w) ointments containing ethanolic extract of Plecranthus amboinicus root was evaluated in terms of Psoriasis severity index (PSI) by the phenotypic features (redness, scales and erythema) and histological features (epidermal thickness). The result showed that there was a significant increase in the orthokeratinocyte layer and a significant reduction in the epidermal layer of skin in the in vivo mice model with a progressive reduction (p**st day and significant (p*Plecranthus amboinicus root. The present investigations revealed that Plecranthus amboinicus root possess potent antipsoriatic activity, confirming their traditional use in skin disorders

The neuroprotective effect of ascorbic acid against imidacloprid-induced neurotoxicity and the role of HO-1 in mice

Imidacloprid (IMI) is not only a neurotoxic agricultural pesticide but also a possible food contaminant. The aims of this study were to (1) explore the relationship between recurrent IMI administration and neuronal toxicity in mice and (2) evaluate the potential neuroprotective effect of ascorbic acid (AA), a substance with significant free radical scavenger and having property to block the inflammatory pathways. Mice were categorized as naïve controls (administered vehicles for 28 days); the IMI-treatment animal group (administered po 45-mg/kg body weight of IMI per day for 28 days); and the IMI + AA treatment animal group (administered the same IMI dose + 200 mg/kg of AA orally for 28 days). On day 28, memory losses were assessed using the Y-maze and novel target identification behavioral tests. Mice were sacrificed 24 h after the final IMI treatments, as well as hippocampus tissues, were utilized to determine histological assessments, oxidative stress biomarkers, and Heme oxygenase-1 (HO-1) and nuclear factor erythroid 2-related factor 2 (Nrf2) gene expression levels. The findings demonstrated that IMI-treated mice had substantial impairment of spatial and non-spatial memory functions, as well as reduced antioxidant enzyme and acetylcholinesterase activity. The AA neuroprotective action was achieved through the suppression of the HO-1 expression as well as the stimulation of Nrf2 expression in hippocampal tissues. In summary, recurrent IMI exposure causes oxidative stress and neurotoxicity in mice, and the administration of AA significantly reduces the IMI toxicity possibly by the activation of the HO-1/Nrf2 pathway