Formulation and Evaluation of Nano Drug Delivery System of Antibiotic Drug Using Factorial
FORMULATION AND EVALUATION OF NANO DRUG DELIVERY SYSTEM OF ANTIBIOTIC DRUG USING FACTORIAL DESIGN
M.Pharm dissertation protocol submitted to
Rajiv Gandhi University of Health Sciences, Karnataka
Bangalore-560041
By
Mr. P. NAGA PAVAN KUMAR B.pharm
Under the Guidance of
Dr. DIVAKAR GOLI M.pharm, Ph.D
Professor & Princical
2010-2011
Department of Pharmaceutics
Acharya & B.M. Reddy College of Pharmacy
Soldevanahalli, Chikkabanavara Post,
Hesarghatta main road, Bangalore-560 090
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
BANGALORE, KARNATAKA.
ANNNEXURE II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1 / Name and address of candidate / Mr. P. NAGA PAVAN KUMAR1734, 22nd MAIN, 41st A CROSS, 4th T BLOCK
JAYANAGAR,BANGALORE
KARNATAKA-560041
2 / Name of institution / ACHARYA & B.M. REDDY COLLEGE OF PHARMACY
SOLDEVANAHALLI, HESARAGHATTA MAIN ROAD,
CHIKKABANAVARA POST.
BANGALORE-560090
3 / Course of study and subject / M.Pharm
(PHARMACEUTICS)
4 / Date of admission / 6th July 2010
5 / Title of the project / FORMULATION AND EVALUATION OF NANO DRUG DELIVERY SYSTEM OF ANTIBIOTIC DRUG USING FACTORIAL DESIGN.
6 / BRIEF RESUME OF THE INTENDED WORK:-
6.1 NEED FOR THE STUDY:
Nanotechnology is defined as the science and engineering carried out in the nanoscale that is 10−9 m. Nanotechnology is a multidisciplinary scientific undertaking, involves creation and utilization of materials, devices, or systems on the nanometer scale. Nanoparticles represent a promising drug delivery system of controlled and targeted release. In this context, nanosuspensions will be effective in increasing the solubility and bioavailability of poorly soluble drugs. Nanosystems with different compositions and biological properties have been extensively investigated for drug and gene delivery applications. Several anticancer drugs, including paclitaxel, doxorubicin, 5-fluorouracil, and dexamethasone, have been successfully formulated using Nanomaterials17.
Poorly soluble drugs are a general problem in pharmaceutical drug formulation and it is expected to increase because approximately 40% or more of the new chemical entities being generated through drug discovery programmes are poorly water-soluble. Typical problems associated with poorly soluble drugs are a too low bioavailability and/or erratic absorption. In case of a too low bioavailability after oral administration, parenteral administration cannot solve this problem in many cases. Hence for the drugs with poor solubility and bioavailability can be improvised by the application of nanotechnology. Size reduction process increases the surface area of drug particles and improves the dissolution rate. The promising approach to transform the drug particles to sub-micron size can be achieved through nano suspensions technology17.
Nanosuspension technology offers novel solution for poorly soluble drugs. Nanosuspensions are submicron colloidal dispersion of pure particles of drug stabilized by surfactants. The average particle size ranges less than 1μm in diameter. A nanosuspension not only solves the problems of poor solubility and bioavailability but also alters the pharmacokinetics of drug and thus improves drug safety and efficacy18. Formulation of Drug nanoparticle by Homogenization includes initial aqueous surfactant solution preparation under magnetic stirring followed by pre-milling low pressure homogenization cycles and finally applied high pressure homogenization7. Samples were withdrawn after the different size reduction steps for size distribution analysis. Lyophilization was applied to in order to retrieve the nanoparticles in dried powder state from nanosuspension.
Advantages:
1. Applicable to the drugs those are poorly soluble in both aqueous and organic media.
2. Ease of scale-up with little batch to batch variation.
3. High flexibility in handling large quantities of drugs.
4. Very diluted as well as highly concentrated nanosuspensions can be prepared by handling
1 to 400 mg/ml drug quantity.
5. Nanosize distribution of final nanosize products17.
Disadvantages:
1. Some fractions of particles are in micrometer range
2. Prerequisite for drug to be in micronized state and suspension formation before homogenization.
3. High-cost instruments.
4. High number of homogenization cycles.
5. Possible contamination of product could occur from metal17.
Formulation of nanoparticles by nano precipitation method includes mixing of organic phase into aqueous phase and followed by organic solvent evaporation under reduced pressure. In this method, organic phase is used to improve the solubility of drug by dissolving a polymer in suitable solvent into which the drug is incorporated. Aqueous phase is prepared by dissolving surfactant/stabilizer in water. Finally the volume of the colloidal suspension is adjusted to meet the required concentration of drug1.
Clarithromycinis amacrolide antibioticact through inhibiting protein synthesis by binding at 50S ribosomal sub-unit and thus inhibits the translation ofpeptides. Used in the treatment ofviral induced asthma, pharyngitis,tonsillitis, acute maxillarysinusitis, acute bacterial exacerbation of chronicbronchitis,pneumonia, skin and skin related infections, etc. It is effective against a broad spectrum of gram-positive and gram-negative bacteria. It is absorbed and diffused easily into tissues and phagocytes without being protected from gastric acids20.
Bioavailability of clarithromycin is 40-50% and half life of about 3-4 hr with low protein binding. It is practically insoluble in water19. These disadvantages facilitate in application of nanosuspension technology to clarithromycin so as to improve its bioavailability, solubility and reduction of dose and side effects. Some of the serious side effects of clarithromycin are:
· Uneven heartbeats, chest pain, shortness of breath;
· Diarrhoea that is watery or bloody;
· Nausea, stomach pain, low fever;
· Loss of appetite, dark urine, clay-colored stools;
· Jaundice (yellowing of the skin or eyes);
· Sore throat and headache with a severe blistering;
· Peeling, and red skin rash or problems with your hearing21.
6.2 REVIEW OF LITERATURE:-
· Martine LLV, et al1, report in this article that isradipine, an antihypertensive agent, was encapsulated by the nanoprecipitation method using polymers including poly (epsilon-caprolactone), poly (d, l-lactide) and poly (d, l-lactide-co-glycolide). In vitro scanning electron microscopy and differential scanning calorimetry were used to characterize the nanoparticles. The average diameters of the nanoparticles ranged from 110 nm to 208 nm. PCL nanoparticles were larger than nanoparticles prepared with the other polymers. The zeta potential of the nanoparticles was negative, with values of about 25 mV which promoted good stabilization of the particles. The amorphous state of PLA and PLAGA non-loaded nanoparticles and the semi-crystalline state of PCL were demonstrated with X-ray diffraction and differential scanning calorimetry. For all nanoparticles, isradipine was found to be totally amorphous in the polymer which suggested that the drug was molecularly dispersed in the matrix. The colloidal suspensions displayed a sustained release profile in comparison with the drug release profile of isradipine in a PEG solution. Results from this investigation suggest that these nanospheres will be a good candidate delivery system for oral administration, to reduce the initial hypotensive peak and to prolong the antihypertensive effect of the drug.
· Hiroyuki O, et al2, report in this article that nifedipine (NI) is a poorly water-soluble drug and its oral bioavailability is very low. To improve the water solubility, NI-lipid nanoparticle suspensions were prepared by a combination of co-grinding by a roll mill and high-pressure homogenization without any organic solvent. The mean particle size and zeta potential of the NI-lipid nanoparticle suspensions were about 52.6nm and −61.8 mV, respectively, and each parameter remained extremely constant during a period of 4 months under 6◦C and dark conditions, suggesting that the negative charge of the phospholipid, dipalmitoyl phosphatidylglycerol, is very effective in preventing coagulation of the particles. In order to assure the nano-order particle size of the suspensions in view of long-term stability, a freeze-drying technique was applied to the NI-lipid nanoparticles suspensions. The mean particle size of freeze-dried NI-lipid nanoparticles after reconstitution was significantly increased in comparison to that of the preparations before freeze-drying. It was found, however, that the addition of sugars (glucose, fructose, maltose or sucrose) to the suspensions before freeze-drying inhibited the aggregation of nanoparticles, suggesting that the long-term stability storage of freeze-dried NI-lipid nanoparticles after reconstitution would be overcome. In addition, freeze-dried nanoparticles with 100mg sugar (glucose, fructose, maltose or sucrose) showed excellent solubility (>80%), whereas without sugar, as a control, showed low solubility (<20%). It was found that negatively charged phospholipids and sugars prevent coagulation of NI nanoparticle suspensions, and reproduce the nanoparticles dispersion after reconstitution and remarkably increase the apparent solubility of nifedipine.
· Gabrielle P, et al3, report in this article that using high-pressure homogenization and spray-drying techniques, novel formulations were developed for manufacturing dry powder for inhalation, composed of a mixture of micro- and nanoparticles in order to enhance lung deposition. Particle size analysis was performed by laser diffraction. Spray-drying was applied in order to retrieve nanoparticles in dried-powder state from tobramycin nanosuspensions. The aerolization properties of the different formulations were evaluated by a multi-stage liquid impinger. Suspensions of nanoparticles of tobramycin containing Na glycocholate at 2% (w/w) relative to tobramycin content and presenting a mean particle size about 200 nm were produced. The results from the spray-dried powders showed that the presence of nanoparticles in the formulations improved particle dispersion properties during inhalation. The fine particle fraction (percentage of particles below 5m) increased from 36% for the raw micronized tobramycin material to about 61% for the most effective formulation. These new nanoparticle containing tobramycin DPI formulations based on the use of very low level of excipient and presenting high lung deposition properties, offer very important perspectives for improving the delivery of drugs to the pulmonary tract.
· Hecq J, et al4, report in this article that ucb-35440-3 is a new drug entity under investigation at UCB S.A. Due to its physicochemical characteristics, the drug, a poorly water soluble weak base, shows poor solubility and dissolution characteristics. In rat, the low oral bioavailability (F < 10%) is largely due to poor absorption. In order to enhance the solubility and dissolution characteristics, formulation of ucb-35440-3 as nanocrystals has been achieved in this study. Nanoparticles were prepared using high pressure homogenization and were characterized in terms of size and morphology. In vitro dissolution characteristics were investigated and compared to the un-milled drug in order to verify the theoretical hypothesis on the benefit of increased surface area. In vivo pharmacokinetic evaluation of ucb-35440-3 nanoparticles was also carried out on rats. Crystalline state evaluation before and following particle size reduction was conducted through polarized light microscopy and PXRD to denote any possible transformation to an amorphous state during the homogenization process. Drug chemical stability was also assessed following homogenization. The dissolution rate increased significantly at pH 3.0, 5.0 and 6.5 for ucb-35440-3 nanoparticles.
However, the pharmacokinetic profile obtained yielded lower systemic exposure than the un-milled compound (in fed state), this although being thought to be the consequence of the drug and formulation characteristics.
· Tao T, et al5, report in this article that the purpose of this work was to explore the feasibility of preparing itraconazole hydrochloride to improve the solubility and dissolution rate. Itraconazole dihydrochloride was synthesized by bubbling anhydrous hydrogen chloride gas into the acetone suspension of itraconazole. Results of the elementary analysis gave the molecular formula of C35H38Cl2N8O4·2HCl and its structure was confirmed by Fourier transform infrared (FTIR), thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC). Powder X-Ray diffraction (PXRD) suggested that a new crystalline form of the salt was formed. The morphology and mean size distribution study by scanning electron microscopy (SEM) and dynamic light scattering (DLS) confirmed that the salt was dispersible nanoparticle aggregation. Aqueous solubility measurements showed that the solubility of the salt, its 1:1, 1:2 and 1:3 (w/w) physical mixtures with beta-cyclodextrin (CD)was 6, 99, 236 and 388 times greater than itraconazole. More than 94% of itraconazole was dissolved out of the salt-CD 1/3 physical mixture after 60 min. The stability studies indicated that the physical mixture remained stable for 24 months in assay, the related substances and dissolution. Based on the present results, it is concluded that hydrochloride formation can significantly increase solubility and dissolution rate of itraconazole, and the formulation of itraconazole dihydrochloride CD (1/3) would be an environment-friendly, economic and practical alternative to the commercially available itraconazole Capsules (Sporanox).
· Lee J, et al6, report in this article that comminution has evolved into an effective method to prepare drug nanoparticles. Although nano-comminution has advantages, such as cost effectiveness and easy scale-up, the processing is significantly sensitive to the selection of a polymeric stabilizer, which suffers from a lack of systematic understanding in this field. Herein, the combinations of various water-insoluble drugs and pharmaceutical polymers were systematically compared to assess the general relationships between the properties of the drugs and polymers. As a rule of thumb, drugs of high molecular weight, low solubility, high melting points, and a surface energy similar to that of the polymers, can be successfully processed into nanoparticles of unimodal particle size distribution. The addition of small molecular weight surfactants results in an additional size reduction in certain polymer/drug pairs, generally by reducing the size of larger particles. Both anionic and cationic surfactants produce similar size reductions in a polymer/drug pair indicating that the charge–charge interaction between polymer and surfactant is not important.
· Hecq J, et al7, report in this article that poorly water-soluble drugs such as nifedipine (NIF) (20g/ml) offer challenging problems in drug formulation as poor solubility is generally associated to poor dissolution characteristics and thus to poor oral bioavailability. In order to enhance these characteristics, preparation of nifedipine nanoparticles has been achieved using high pressure homogenization. The homogenization procedure has first been optimized in regard to particle size and size distribution. Nanoparticles were characterized in terms of size, morphology and redispersion characteristics following water-removal. Saturation solubility and dissolution characteristics were investigated and compared to the un-milled commercial NIF to verify the theoretical hypothesis on the benefit of increased surface area. Crystalline state evaluation before and following particle size reduction was also conducted through differential scanning calorimetry (DSC) and powder X-ray diffraction (PXRD) to denote eventual transformation to amorphous state during the homogenization process. Through this study, it has been shown that initial crystalline state is maintained following particle size reduction and that the dissolution characteristics of nifedipine nanoparticles were significantly increased in regards to the commercial product.