Curriculum Development Cell
CONFIRMATION FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
Registration No. / :
Name of the Candidate / : Mohd Mohiuddin Ahmed
Address / : No -79 , Mariyaman koil st , Thattanchavdy, Pondicherry - 605009
Name of the Institution / : Al - Ameen College of Pharmacy, Bangalore
Course of Study and Subject / : M . Pharm in Pharmaceutics
Date of Adimission to Course / :
Title of the Topic / : Formulation and evaluation of nanoparticles for oral delivery of Artemether
Brief resume of the intended work / : Attached
Signature of the Student / :
Guide Name / : Dr. (Mrs.) V. KUSUM DEVI
Remarks of the Guide / :
Signature of the Guide / :
Co-Guide Name / : Dr. (Mrs.) ASHA A.N
Signature of the Co-Guide / :
HOD Name / : Dr. (Mrs.) V. KUSUM DEVI
Signature of the HOD / :
Principal Name / : Prof. B.G.Shivananda
Principal Mobile No. / : 09844023000
Principal E-mail ID / :
Remarks of the Principal
/ : Recommended
Principal Signature / :
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA
4TH ‘T’ BLOCK, JAYANAGAR, BANGALORE - 560 041
ANNEXURE – II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. / Name of the Candidate and Address / MOHD MOHIUDDIN AHMEDNo : 79 , Mariyaman koil st ,
Thattanchavdy,
Pondicherry – 605009 , INDIA.
2. / Name of the Institution / AL-AMEEN COLLEGE OF PHARMACY,
HOSUR ROAD,
OPP. LALBAGH MAIN GATE,
BANGALORE – 560 027.
KARNATAKA.
3.
4. / Course of Study and Subject
Date of Admission / M. PHARM – PHARMACEUTICS
JUNE-2010
5. / Title of the Topic:
“ FORMULATION AND EVALUATION OF NANOPARTICLES FOR ORAL DELIVERY OF ARTEMETHER ”
6.0 / Brief resume of the intended work:
6.1 – Need for the study
Parasitic diseases are of immense global significance as around 30% of world’s population experiences parasitic infections . Amongst various parasitic infections, malaria is the most life threatening disease with 300–500 million clinical cases each year and accounts for 1 million to 2 million deaths round the globe every year. About two thirds of reported cases in the region are from India which reported 1.5 million confirmed cases in 2008 down from 2 million in 2000. The percentage of cases due to P.falciparum has fluctuated from between 44% and 51% . 1,2
In humans, malaria is caused by four distinct species of parasites: Plasmodium vivax, Plasmodium falciparum, Plasmodium malariae and Plasmodium ovale.Amongst these, the most severe malaria is caused by blood-borne Apicomplexan parasite P. falciparum which is responsible for almost all malaria related deaths. 1
Artemether is a potent and rapidly acting anti-malarial drug which is listed by WHO as an Essential Drug for the treatment of severe multi-resistant malaria. This potent schizonticide drug is practically insoluble in water, belongs to BCS class II and has oral bioavailability of 45%. The generally recommended oral and parenteral administration, once a day for at least 5 days seems reasonable in view of clinical efficacy. 3
The artemisinin derivative artemether contains sesquiterpene lactone rings with an endoperoxide bridge that is cleaved by an iron-dependent mechanism, playing a crucial role in the treatment of multi-resistant malaria parasite. Artemether has been found to inhibit haemozoin formation as well as haemoglobin degradation, due to the presence of the haem group that is a potent inhibitor of cysteine protease. It inhibits PfATPG outside the parasite’s food vacuole and has fast and potent action in blood stages and shows gametocytocidal effect.1
However, this drug holds some important shortcomings, including (i) short half-life usually between 3h and 5h; (ii) poor aqueous solubility, and thus low oral bioavailability (40%); (iii) risk of degradation in acidic conditions; and (iv) associated risk of toxicity. Intramuscular (i.m.) injections currently available in the market are associated with low patient compliance and inconsistent assimilation. 4
The marketed dosage forms available for Artemether are tablet, powder filled capsule and intramuscular injection. The oral formulations of Artemether are rapidly but incompletely absorbed, limiting its use in malaria , Karbwang et al., 1997 J. Karbwang, K. Na-Bangchnag, K. Congpoung, P. Molunto and A. Thanavibul, Pharmacokinetics and bioavailability of oral and intramuscular Artemether, Eur. J. Clin. Pharmacol. 52 (1997), pp. 307–310. Full Text via CrossRef | View Record in Scopus | Cited By in whereas parenteral oily formulation leads to pain on injection and poor patient compliance. 5
The emergence of drug-resistant parasite strains indicates an urgent need for discovering new and effective anti-malarial therapeutics as well as effective utilization of existing drugs through the concept of novel drug delivery systems. 6
Nanocarriers as drug delivery systems is to promote drug or vaccine protection against extracellular degradation, to improve selectivity in relation to the target, to reduce the frequency of administration and the duration of the treatment and to improve the pharmacokinetic profile of the drug. Nanoparticulate drug delivery systems represent a promising approach for obtaining desirable drug like properties by altering the biopharmaceutics and pharmacokinetics property of the drug molecule. 1
In order to overcome the shortcomings of artemether and conventional drug delivery , nanoparticles as carriers can be used for artemether.
The most important property of a nanocarrier in the context of malaria is the ability to remain in the blood stream for a long period of time in order to improve the interaction with infected red blood cells (RBCs) and parasite membranes. 1
/ 6.2 REVIEW OF LITERATURE
Ø Artemether oil soluble methyl ether is highly effective against the blood schizonts of both malarial parasites P. falciparum and P. vivax. Peak plasma concentrations have been achieved in about 3 hours after oral doses of artemether , in about 6 hours after i.m injection of artemether. Reported elimination half-lives have been about 4 to 11 hours after intramuscular or oral artemether. Artemether appears to be generally well tolerated , although there have been reports of mild GI disturbances , dizziness , headache , tinnitus , neutropenia , and ECG abnormalities including prolongation of QT interval. 7
Ø Artemether is rapidly and extensively metabolized with substantial first-pass metabolism. Artemether is metabolized in the liver to the biologically active main metabolite Dihydroartemisinin (demethylation) , predominantly through the isoenzyme CYP3A4/5. 8
Ø Artemether with its endoperoxide bridge appears to interact with haeme in the parasite. Iron-mediated cleavage of the of the bridge releases a highly reactive free radicals species that binds to membrane proteins , cause lipid peroxidation , damages endoplasmic reticulum , inhibits protein synthesis and ultimately results in lysis of the parasite. 9
Ø Sagar D. Mandawgade et al. developed β-artemether SMEDDS using a novel, indigenous natural lipophile (N-LCT) as an oily phase and commercially available modified oil (Capryol 90) in order to improve the delivery of a lipophilic anti-malarial drug. SMEDDs is an isotropic, anhydrous mixture of drug, lipophile and surfactant/s that form fine oil-in-water microemulsion (globule size <100nm) when introduced into aqueous phase under conditions of gentle agitation. SMEDDS are used because of high drug solubilizing capacity and improvement in both rate and extent of absorption by the lymphatic uptake. β-Artemether (BAM) was also evaluated. BAM-loaded SMEDDS were characterized with respect to mean globule size and in vitro drug release profile in comparison to the marketed formulation (Larither®). Comparative in vivo anti-malarial performance of the developed SMEDDS was evaluated against the (Larither®) in Swiss male mice infected with lethal ANKA strain of Plasmodium berghei. β- artemether SMEDDS showed excellent self-microemulsification efficiency and released >98% of the drug in just 15mins and mean globule size was <100nm. The anti-malarial studies revealed that BAM–SMEDDS resulted in significant improvement in the anti-malarial activity (P0.05) and BAM solubilized in the oily phases and surfactant. The developed SMEDDS highlight safety for use and potential applications of indigenous natural lipophile in the development of novel colloidal drug carriers. 3
Ø Aditya N.P et al. developed artemether-loaded lipid nanoparticles produced by a modified thin-film hydration method using glyceryl trimyristate and soybean oil. The particles were loaded with 10% of the anti-malarial ARM and surface-tailored with a combination of non-ionic, cationic or anionic surfactants. ARM-LNP were further characterized for their mean particle size, zeta potential and encapsulation efficiency, reporting optimized values below 120nm (PI0.250), −38mV and 97% (w/w), respectively. ARM-LNP composed of 45% soybean oil depicted a spherical-like shape by transmission electron microscopy and a biphasic release profile in phosphate buffer. In vivo anti-malarial activity of ARM was enhanced when formulated as LNP, in comparison to a conventional plain drug solution and to a marketed formulation which are currently in use to treat malaria patients. 4
Ø Magnus A. Atemnkeng et al. described two HPLC-UV methods for the separate determination of artemether (AM) and the combined preservatives, methylparaben and propylparaben in a pharmaceutical dosage form. Artemether was analysed using a reversed-phase Nucleosil® C18 column [5μm, 125mm×4mm (i.d.)] with a mixture of acetonitrile: potassium phosphate buffer pH 5.0 (0.05M): water [48:32:10 (v/v/v)] as mobile phase. Quantitation was achieved with UV detection at 215nm for artemether and 254nm for the parabens, respectively. The procedures prescribed here are simple, selective and can be used for routine quality control and stability indicating tests involving the analysed compounds formulated in complex matrices. 10
Ø Chimunaka B et al. prepared egg phosphatidylcholine-cholesterol liposome formulations containing artemether and analyzed for their encapsulating capacity, chemical stability, leakage, in vitro release and their therapeutic efficiency against Plasmodium chabaudi infection. A HPLC–UV analysis of β- artemether liposomes using acetonitrile : water (75 : 25) as mobile phase without derivatisation was achieved. A good linearity of y=4437.7x+469.01 (R2=0.9999) with a detection limit of 2 μg ml−1 was reached. A trapping efficiency of nearly 100% was reached, the drug being located in the lipid bilayers. After 3 months storage at 4 °C, no leakage of β -artemether had occurred indicating a high stability of the liposomes. These liposomes were used to treat mice infected with the virulent rodent malaria parasite Plasmodium chabaudi chabaudi, with a 100% cure by clearing the recrudescent parasitaemia. 11
Ø Medha Joshi et al. formulated solid microemulsion preconcentrate (NanOsorb) and it significantly improved the therapeutic efficacy of artemether demonstrating the utility of the novel drug delivery strategies for antimalarial agents. In vivo studies clearly demonstrated that NanOsorb-ARM has significantly higher antimalarial activity as compared to the ARM solution and marketed formulation (Larither®). The acute and subacute toxicity studies successfully established the safety of the NanOsorb in the animals. 12
Ø Bhadra D et al . solubilized artemether in PEG-lysine type dendritic peptide based nanoparticulate carriers for sustained and controlled delivery of the drug through i.v route of administration. The systems were further conjugated with chondritin sulphate A. The systems were found to be entrapping 10-18 molecules of ART per molecule of dendrimer. The release was prolonged up to 1-2 days. Conjugation with CSA increased amount of drug loading and prolonged drug release by 2 to 3 folds. 13
Ø Medha Joshi et al. formulated nanostructured lipid carriers ( NLC ) of artemether by employing a microemulsion template technique.The average particle size of Nanoject was 63 ± 28 nm and the encapsulation efficiency was found to be 30±2%. The Nanoject released ARM in a sustained manner. The antimalarial activity of Nanoject lasted for a longer duration (more than 20 days) indicating that Nanoject may be long-circulating in vivo. 14
Ø Manoj Nahar et al. developed a nanoparticulate carrier of amphotericin B for controlled delivery as well as reduced toxicity. Nanoparticles of different gelatins (GNPs) (type A or B) were prepared by two-step desolvation method and optimized for temperature, pH, amount of cross-linker, and theoretical drug loading. AmB-loaded GNPs were characterized for size, polydispersity index (PI), shape, morphology, surface charge, drug release, and hemolysis. The developed GNPs (GNPA300) were found to be of nanometric size (213 ± 10 nm), having low PI (0.092 ± 0.015) and good entrapment efficiency (49.0 ± 2.9%). All GNPs showed biphasic release characterized by an initial burst followed by controlled release. The in vivo hematological toxicity results suggest nonsignificant reduction (P > .05) in hemoglobin concentration and hematocrit. Nephrotoxicity results showed that there was a nonsignificant (P > .05) increase in blood urea nitrogen and serum creatinine levels. The results confirm that developed GNPs could optimize AmB delivery in terms of cost and safety, and type A gelatin with bloom number 300 was found suitable for such preparation. 15
Ø Liyan Qiu et al. developed active loading method for encapsulating chloroquine diphosphate into liposomes. The effect of different formulation factors on the encapsulation efficiency (EE) and the size of CQ liposomes were investigated. The EE (93%) was obtained when using drug/SPC (1:50 mass ratio), SPC/Chol (1:5 mass ratio) at 0.10M citrate–sodium citrate buffer (pH 3.6). As 5mol% methoxypoly(ethylene glycol)2000 cholesteryl succinate (CHS-PEG2000) or distearoyl phosphatidylethanolamine-poly (ethylene glycol)2000 (DSPE-PEG2000) was added, the size of particle was reduced and the EE was improved. Freeze-drying with 5% trehalose as a cryoprotectant was carried out to achieve long-term stability. The drug release studies were performed in vitro simulating the desired application conditions, such as physiological fluids (pH 7.4), tumor tissues (pH 6.5) and endosomal compartments (pH 5.5). The release of CQ from the liposomes
prepared via remote loading showed the significant pH-sensitivity and retention properties, which favored the application of liposomal CQ at tumor tissues and endosomal compartments. 16
Ø Padma V. Devarajan et al. formulated and optimize gliclazide-loaded Eudragit nanoparticles (Eudragit L100 and Eudragit RS) as a sustained release carrier with enhanced efficacy. Eudragit L 100 nanoparticles (ELNP) were prepared by controlled precipitation method whereas Eudragit RSPO nanoparticles (ERSNP) were prepared by solvent evaporation method. Mean particle size altered by changing the drug:polymer ratio and stirring speed. Addition of surfactants showed a promise to increase drug loading, encapsulation efficiency, and decreased particle size of ELNP as well as ERSNP. Dissolution study revealed sustained release of gliclazide from Eudragit L100 as well as Eudragit RSPO NP. SEM study revealed spherical morphology of the developed Eudragit (L100 and RS) NP. FT-IR and DSC studies showed no interaction of gliclazide with polymers. Stability studies revealed that the gliclazide-loaded nanoparticles were stable at the end of 6 months. 17