Satish Shilpi

Eudragit S100 Coated Citrus Pectin Nanoparticles for Colon Targeting of 5-Fluorouracil

In the present study, Eudragit S100 coated Citrus Pectin Nanoparticles (E-CPNs) were prepared for the colon targeting of 5-Fluorouracil (5-FU). Citrus pectin also acts as a ligand for galectin-3 receptors that are over expressed on colorectal cancer cells. Nanoparticles (CPNs and E-CPNs) were characterized for various physical parameters such as particle size, size distribution, and shape etc. In vitro drug release studies revealed selective drug release in the colonic region in the case of E-CPNs of more than 70% after 24 h. In vitro cytoxicity assay (Sulphorhodamine B assay) was performed against HT-29 cancer cells and exhibited 1.5 fold greater cytotoxicity potential of nanoparticles compared to 5-FU solution. In vivo data clearly depicted that Eudragit S100 successfully guarded nanoparticles to reach the colonic region wherein nanoparticles were taken up and showed drug release for an extended period of time. Therefore, a multifaceted strategy is introduced here in terms of receptor mediated uptake and pH-dependent release using E-CPNs for effective chemotherapy of colorectal cancer with uncompromised safety and efficacy.

Development of surface-functionalised nanoparticles for FGF2 receptor-based solid tumour targeting.

The surface-functionalised gelatin nanoparticles (GNPs) containing cisplatin were developed and characterised for breast cancer targeting using fibroblast growth factor-2 (FGF2) receptors which are overexpressed on breast cancer cells. The GNPs were prepared using two-step desolvation method and then the surface of GNPs was functionalised with activated heparin. They were characterised for surface morphology, particle size and size distribution, surface charge, entrapment efficiency and in vitro drug release. The results revealed that the mean diameter of GNPs was 173 ± 2.2 nm with smooth surface, which was increased to 189 ± 3.4 nm after coupling with heparin (H-GNPs). The targeting effect of H-GNPs and GNPs was investigated by in vitro cell uptake study on human breast cancer MDA-MB-231 cell line, which exhibited greater uptake of H-GNPs as compared to GNPs. Therefore, it is suggested that H-GNPs can be used as an effective carrier for solid tumour targeting.

Mannosylated liposomes bearing Amphotericin B for effective management of visceral Leishmaniasis

The cationic and mannosylated liposomes were prepared using the cast film method and compared for their antileishmaniasis activity. The surface of the Amphotericin B (Amp B)-bearing cationic multilamellar liposomes was covalently coupled with p-aminophenyl-α-D-mannoside using glutaraldehyde as a coupling agent, which was confirmed by agglutination of the vesicles with concanavalin A. The prepared liposomes were characterized for shape, size, percent drug entrapment, vesicle count, zeta potential, and in vitro drug release. Vesicle sizes of cationic and mannosylated liposomes were found to be 2.32 ± 0.23 and 2.69 ± 0.13 µm, respectively. Zeta potential of cationic liposomes was higher (30.38 ± 0.3 mV), as compared to mannosylated liposomes (17.7 ± 0.8 mV). Percentage drug release from cationic and mannose-coupled liposomes was found to be 45.7% ± 3.1 and 41.9% ± 2.8, respectively, after 24 hours. The in vivo antileishmanial activity was performed on Leishmania donovani–infected golden hamster, and results revealed that Amp B solution was reduced by 42.5 ± 1.8% in the parasite load, whereas the placebo cationic liposomes and drug-containing cationic liposomes showed a reduced parasite load (i.e., 28.1 ± 1.5 and 61.2 ± 3.2%, respectively). The mannose-coupled liposomes showed a maximum reduction in parasite load (i.e., 78.8 ± 3.9%). The biodistribution study clearly showed the higher uptake of mannosylated liposomes in the liver and spleen and hence the active targeting to the reticular endothelial system, which, in turn, would provide a direct attack of the drug to the site where the pathogen resides, rendering the other organs free and safe from the toxic manifestations of the drug.

Targeting liver cancer via ASGP receptor using 5-FU-loaded surface-modified PLGA nanoparticles

Abstract Context: Liver cancer is widespread liver malignancy in the world, for an estimated one million deaths annually. Objective: In present work, lactobionic acid conjugated PLGA nanoparticles (LDNPs) bearing 5-Fluorouracil (5-FU) were developed for targeted delivery to hepatocellular carcinoma. Materials and methods: Lactobionic acid conjugated PLGA was used to prepare LDNPs using modified emulsion diffusion method. Results: They were characterised for particle morphology, particle size (below 150 nm), zeta potential and polydispersity index (PDI ∼0.35), entrapment efficiency (∼60.23%), and cumulative percent drug release. Discussion: LDNPs in ex-vivo cell line studies on human cancer cell line HepG2 exhibited significantly higher cytotoxicity compared to 5-FU and DNPs (unconjugated PLGA NPs) with growth inhibition 50% (GI50) of 66.7 µg/mL, 50.2 µg/mL and 35.5 µg/mL, respectively. In vivo studies exhibited higher drug concentration about 37.52 ± 0.68% in liver as compared to other organs and plasma. Conclusion: Thus, LDNPs showed high drug loading, specificity, biocompatibility and efficacy in treatment of liver cancer.

Insulin delivery through nasal route using thiolated microspheres

Abstract The aim of the present study was to investigate the potential of developed thiolated microspheres for insulin delivery through nasal route. In the present study, cysteine was immobilized on carbopol using EDAC. A total of 269.93 µmol free thiol groups per gram polymer were determined. The prepared nonthiolated and thiolated microspheres were studied for particle shape, size, drug content, swellability, mucoadhesion and in vitro insulin release. The thiolated microspheres exhibited higher mucoadhesion due to formation of covalent bonds via disulfide bridges with the mucus gel layer. Drug permeation through goat nasal mucosa of nonthiolated and thiolated microspheres were found as 52.62 ± 2.4% and 78.85 ± 3.1% in 6 h, respectively. Thiolated microspheres bearing insulin showed better reduction in blood glucose level (BGL) in comparison to nonthiolated microspheres as 31.23 ± 2.12% and 75.25 ± 0.93% blood glucose of initial BGL were observed at 6 h after nasal delivery of thiolated and nonthiolated microspheres in streptozotocin-induced diabetic rabbits.

Transferrin conjugated nanoparticles for intracellular delivery of anticancer drug

T aim of current study entails to develop a solid lipid nanoparticle (SLN) as a novel lipid nanocarrier for the oral delivery of decitabine (DCB) using cold homogenization technique. A Box-Behnken design (33) with 17 experimental runs was constructed to identify the key independent variables influencing on dependent variables. The optimized batch (SLN-11) was further characterized with particle size distribution, zeta potential, TEM, entrapment efficiency, drug content, rheological study, DSC, in vitro drug release, and accelerated stability. The optimized batch revealed spherical morphology under TEM analysis with particle size of 136.6± 2.35 nm and 0.244±0.002 PDI. Zeta potential and %EE was found to be -31.34±0.67 mV and 58.89% ±0.78 respectively. In vitro release studies showed burst release at the initial stage followed by sustained release up to 24 hrs in intestinal medium and the data was further studies using release kinetic models which revealed Higuchi matrix as a best fitted model. Finally, SLN prepared using Precirol ATO5 as solid lipid and surfactants as Poloxamer 188, Tween 80 and Solutol HS15 (2:1:2 ratio) posses high potential to entrap DCB in lipid nanoparticle, showed better prospects for the oral delivery of DCB.C (cur), a natural compound elicit a spectrum of potent responses both locally and systemically. However its local effect in buccal conditions is largely hindered by its extremely limited water solubility, and its hydrolytic degradation in salivary pH. The aim of the present study was to develop buccal mucoadhesive tablets of cur with accepted release and stability at salivary pH as well as to design a simple in vitro dissolution test ensuring its stability. Chemical stability in phosphate buffer saline (PBS) pH 6.8 was tested using a group of stabilizers of which sodium lauryl sulfate (SLS) proved to be the most suitable. Different muccoadhesive tablets formulations were prepared by direct compression technique using a mixture of Hydroxypropyl methylcellulose (HPMC) K15M and Carboxymethylcelluose sodium (NaCMC) in different ratios with or without SLS as stabilizer, curas pure untreated drug or in the form of rapidly dissolving solid dispersion (SD) with PVP (Kollidon®25). Formulations were evaluatedfor mucoadhesive strength, in vivo and in vitro residence time, release studies and clinical evaluation of the selected formulation. The best mucoadhesive performance and in vitro sustained release profile (70% released over 12 hours) were exhibited by tablets containing HPMC.K15M: CMC sodium (5:1), SD (1:3) with 15 mg SLS. Salivary concentration (conc.) was significantly increased compared to undetectable conc. for pure cur due to poor solubility and SD without SLS due to hydrolytic degradation. Preliminary clinical study revealed an excellent anti-inflammatory and healing effect. Cur in this delivery system is an excellent candidate for local buccal delivery.T purpose of present study is to formulate SNEDDS of BCS Class-IV (Exemestane HCl) to investigate its potential oral drug delivery system by improving its bioavailability. Preformulation study was done for selection of oils, surfactants & co-surfactants. Based on the solubility studies, Caprol microexpress and Labrafac as oil phase, Tween 80 as a surfactant and Triacetin as a co-surfactant were selected. Phase studies were performed using different ratio like (1:1, 1:2, 1:3, 2:1, 3:1) [oil: (surfactant/co-surfactant)]. Pseudo ternary phase diagram were prepared, Tween 80: triacetin (1:2) and (1:3) ratio showed the highest area for the preparation for the nanoemulsion. All formulations were evaluated for the visual assessment, optical clarity, particle size, drug content, viscosity, in vitro release study. From vitro characterization results, three formulations were selected as potential formulation for in vitro cytotoxicity screening and in vivo pharmacokinetic study. EX1 showed particle size (29.56 nm), Polydispersity index (0.523), Zeta potential (-40.3), & drug release after 120 min. was 99.589±1.85 % EX2 showed particle size (37.65 nm ), Polydispersity index (0.835), Zeta potential (-30.3), & drug release after 120 min was 99.17±1.81 % EX3 showed particle size (44.73 nm), Polydispersity index (0.679), Zeta potential (-15.7), & drug release after 120 min was 98.172±1.29 % due to its low particle size and excellent stability. The dose response curves demonstrated that Exemestane SNEDDS had less cytotoxicity compared to the drug solution alone after 24 hrs but EX2 showed greater % cell inhibition as well as greater AUC Compare to EX3 and EX1. It can be concluded that SNEDDS is a novel and commercially feasible approach to improve oral bioavailability of BCS class-IV drug like exemestane HCL.T aim of present study was the development of hyaluronic acid (HA)-targeted pH-sensitive liposomes (SL-pH-HA) for intracellular delivery of doxorubicin (DOX) in the cancer cell that express CD44. The in vitro release studies demonstrated that the release of DOX from SL-pH-HA was pH-dependent, i.e., faster at mildly acidic pH ~5, compared to physiological pH ~7.4. SL-pH-HA was evaluated for cytotoxicity potential on MCF-7 cells. The quantitative uptake study by flow cytometry revealed higher localization of targeted liposomes in the receptor positive cells, which was further confirmed by fluorescent microscopy. The in vivo antitumor activity in tumor bearing mice was also confirmed the efficacy of HA targeted pH-sensitive liposomes. The major side-effect of DOX i.e., cardiotoxicity was also estimated by measuring serum enzyme level. Thus, HA targeted pH-sensitive liposomes were significantly more potent than the non-targeted liposomes in cells expressing high levels of CD44, which suggests that they may be a useful targeted drug carrier to treat CD44-expressing breast tumors.Introduction: Warfarin is the most widely used anticoagulant in the world. The difficulty of managing warfarin contributes to great potential for patient harm, both from excessive anticoagulation and insufficient anticoagulation. Objective: To assess the INR control outcome measures for warfarin therapy, to investigate quality of warfarin prescribing and to assess level of patients’ knowledge on warfarin therapy among outpatients in Tikur Anbessa Specialized Hospital (TASH). Methodology: The study was based on cross sectional study design involving retrospective chart review and Oral Anticoagulation Knowledge Assessment (OAKA) questionnaires. 360 patients’ charts were included and 130 patients were interviewed in this study. Result: Patients spent 52.2%, 29% and 18.8% of the times in subtherapeutic, therapeutic and supratherapeutic ranges, respectively. The daily warfarin dose was increased in (50.9% and 36.9%) and decreased in (52.8% and 60.9%) of the times for occurrences of subtherapeutic and supratherapeutic International Normalized Ratio (INRs) to achieve target ranges of 2.0-3.0 and 2.5-3.5 respectively. The majority of patients (76.9%) had moderate knowledge on warfarin therapy. The mean score of patients on correct response was 11.8±2.5 (59.3%± 12.8%). Among interviewed patients only 13.9% of them got passing scores. Conclusion and Recommendation: This study found that quality of anticoagulation management with warfarin among outpatients in TASH was suboptimal. This was reflected by low Time in Therapeutic Range, longer INR monitoring frequency, minimal actions taken to adjust warfarin dose after occurrences of nontherapeutic INRs; and poor patients’ knowledge. Establishing pharmacist managed anticoagulation clinic which supported by computer software programs may be the integral part of anticoagulation management services in TASH.Materials and Methods: Nanoarticles were prepared in two steps. Firstly biodegradable amphiphilic polymer i.e., poly (H2NPEGCA-co-HDCA) was synthesized which is having high encapsulation efficiency. In second step, doxorubicin and peptide encapsulated NPs of synthesized polymer was prepared using double emulsification method reported by Stella (2000) which was further conjugated with transferrin as reported by Minghuang (2010). The optimized NPs formulations such as NpfDox, Npf-Dox-Tf, Npf-Dox-Tf-Ga respectively were used for further studies. Size and size distribution of NPs were determined using a Zetasizer (Malvern Instruments, UK). Shape and surface morphology of NPs were studied using TEM and SEM. In vitro drug release was carried using dialysis bag (MW 1200 da). Ex vivo antitumor studies such as MTT cell cytotoxicity and cell uptake study were carried out on MCF-7 human breast cancer cell line.HE POSSIBLE anti-inflammatory, cytotoxic and hepatoprotective activities of Laurus nobilis L. wood extract was in vitro evaluated. The anti-inflammatory activity was assessed by measuring Nitric Oxide (NO) production by the inflammagen lipopolysacchride. The extract was evaluated for its cytotoxic activity on six different human cancer cell lines together with the normal non-malignant melanocytes cell line (HFB4) using the SRB assay. The hepatoprotective activity against paracetamol toxicity was determined using primary cultured rat hepatocytes. The extract showed moderate anti-inflammatory activity as shown in the amount of nitric oxide produced with a level of 4.3 μM /ml (67 % inhibition), in comparison to the potent anti-inflammatory drug Dexamethasone (95 % inhibition). The extract was found to have moderate cytotoxic activity against the tumor cell lines used at the applied concentrations with IC50 ranging from 15.5 – 47.6 μg/ml compared to the potent cytotoxic drug doxorubicin. The wood extract showed hepatoprotective activity against paracetamol toxic effect at concentration of 20g/ ml. The constitutive phenolics of L. nobilis L. wood were studied and led to the separation and identification of 12 compounds, all isolated for the first time from L. nobilis L. wood and were identified using chemical, conventional and advanced spectral techniques.An oral press-coated tablet was prepared by using direct compression and wet granulation methods to achieve the predetermined lag time. This press-coated tablet containing montelukast sodium in the inner core was formulated with an outer barrier layer by different compositions of hydrophobic polymer ethylcellulose and hydrophilic polymer low-substituted hydroxypropylcellulose. The effect of formulation composition on the barrier layer comprising both hydrophobic and hydrophilic excipients on the lag time of drug release was investigated. It was observed that lag time decreases with increasing concentration of low-substituted hydroxypropylcellulose. Press coated tablets coated by dry mixing and by wet granulation showed variations in lag time. As compared to dry mixed blend method wet granulation method gives less lag time.Oral salbutamol sulphate has site-specific absorption in the stomach and upper part of the small intestine. Its bioavailability is about 40% due to several factors including narrow absorption window and extensive intestinal metabolism. The aim of this study was to formulate and optimize sustained release floating tablets of salbutamol sulphate in order to improve its bioavailability and reduce its dosing frequency. Accordingly, floating tablets were prepared by wet granulation technique and drug release analysis was performed by HPLC. The effects of polymer level, polymer type (XG or HPMC), polymer ratio (XG/HPMC; 1:1, 1:3, 3:1) and NaHCO3 level on floating lag time, floating duration, cumulative release within 1 hr, and release rate were investigated. From preliminary studies, the polymer with 1:3 (XG:HPMC) ratio and NaHCO3 were selected as significant factors and cumulative release at 1 hr and release rate were chosen as significant responses, respectively. Hence, the effect of these factors were further studied and optimized by central composite design. The most desirable representative optimal point was obtained at 24.79% of XG/HPMC and 5% of NaHCO3 having release rate of 28.49 hr -1/2 and cumulative release at 1 hr of 24%. This formulation is expected to significantly improve bioavailability of salbutamol while remaining buoyant and sustained release INTRODUCTION: Salbutamol sulphate is one of the widely used drugs in the treatment of respiratory disorders like bronchial asthma, chronic bronchitis and obstructive airway diseases 1 . The relatively short acting injectable and aerosol dosage forms of salbutamol sulphate are recommended for instant relief in severe asthmatic attacks. QUICK RESPONSE CODE DOI: 10.13040/IJPSR.0975-8232.6(5).1877-92 Article can be accessed online on: www.ijpsr.com DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.6(5).1877-92 The recommended dose of aerosols in adults and children is 2 – 3 inhalations every 4 – 6 hr 2 and for conventional tablets, 2-6 mg (base) is administered three to four times a day 3 which causes poor patient compliance, multiple administration associated side effects, and plasma drug level fluctuation. Salbutamol sulphate has oral bioavailability of only ~40% due to extensive metabolism via intestinal sulphonation, first pass metabolism in the liver, narrow absorption window (site-specific absorption in stomach and upper part of small intestine 4 and degradation in colon 5, 6 . Hence, development of