formulation and evaluation of hesperidin-loaded ?· preparation of hesperidin nanosuspension and...
Post on 07-Sep-2018
Embed Size (px)
279Drug Invention Today | Vol 10 Issue 3 2018
Formulation and evaluation of hesperidin-loaded chitosan nanosuspension for brain targetingI. Somasundaram*, S. Sumathi, S. P. Bhuvaneshwari, K. Mohammed Shafiq, T. S. Shanmugarajan
INTRODUCTIONThe Parkinsons disease is more prevalent in person with about 65 years of age concomitant with genetic influence, toxins, oxidative stress, mitochondrial abnormalities, free radicals, and alpha-synuclein aggregation in a higher rate, which is considered to be the main cause for Parkinson disease. It is the second most prevalent neurodegenerative extrapyramidal motor disorder succeeding Alzheimers disease. Mostly the cause is idiopathic. Mutation in chromosome 4 and 6 is the main origin for Parkinsonism.[1,2] Dopamine is a neurotransmitter which is feasible in performing the functions of locomotion, learning, working, memory, cognition, and emotion. These functions are performed by binding to their specific receptors which are the members of large G-protein-coupled receptor. The
Department of Pharmaceutics, School of Pharmaceutical Sciences, Vels Institute of Science, Technology and Advanced Studies, Chennai, Tamil Nadu, India
*Corresponding author: I. Somasundaram, Department of Pharmaceutics, School of Pharmaceutical Sciences, Vels Institute of Science, Technology and Advanced Studies Pallavaram, Chennai - 600 117, Tamil Nadu, India. Phone:+91-44-22662500/2501/2502/2503. Fax: +91-44-22662513. E-mail: firstname.lastname@example.org
Received on: 25-09-2017; Revised on: 22-10-2017; Accepted on: 28-01-2018
Access this article online
Website: jprsolutions.info ISSN: 0975-7619
deregulation of the dopaminergic system has a correlation with Parkinsons disease, generation of pituitary tumors, Tourettes syndrome, schizophrenia, and attention deficit hyperactive disorder.[3,4]
Oxidation is the main process in augmenting the energy production. However, oxidative stress is the predictable main cause which enhances the progression of disease. This condition arises when there is a reduction in antioxidant defenses or overproduction in reactive oxygen species (ROS). ROS is generated at the time of injury or inflammation. There are several other sites for ROS production such as mitochondrial electron transport, peroxisomal fatty acid, cytochrome p-450, and phagocytic cells. In low concentration, it serves as signaling intermediates for growth, whereas in high concentration, it induces cell injury and death. A free radical is an atom or group with at least one unpaired electron. It is usually an antioxidant molecule that has lost and needs to stabilize itself by stealing an electron from a nearby molecule. Antioxidant can
Aim: Parkinsons disease is characterized by a progressive loss of dopaminergic neuron in the substantia nigra. Hesperidin is a free radical scavenger that has been shown to have a protective effect against oxidative insults. An antioxidant can neutralize the free radical by accepting or donating an electron to eliminate the unpaired condition. Materials and Methods: Hesperidin-loaded chitosan nanosuspension was prepared by ionic gelation method using tripolyphosphate and chitosan. There was no incompatibility observed between the drug and polymer through Fourier-transform infrared and differential scanning calorimetric. Various other parameters such as particle size, zeta potential, scanning electron microscope, drug content, drug entrapment efficiency, and in vitro release have been utilized for the characterization of nanoparticles. Results and Discussion: The average particle size of 188 nm, zeta potential of 48.7 mV, drug content of 0.470 0.25 mg/ml, and entrapment efficiency of 78.2% are obtained with HPN3 formulation. The HPN1 shows the highest drug release followed by HPN2 due to the low concentration of polymer, and HPN4 and HPN5 show less drug release due to the high concentration of polymer. The in vitro release of HPN3 85.2% showed an initial burst release of approximately 60% in the 1
st 8 h, followed by a slow and much reduced additional release for about 24 h. Conclusion: HPN3 was chosen as the best formulation due to its reduced particle size and controlled release at optimum polymer concentration which may be taken for the effective treatment of Parkinsons disease.
KEY WORDS: Chitosan, Hesperidin, Nanoparticles, Parkinsons disease, Reactive oxygen species
I. Somasundaram, et al.
Drug Invention Today | Vol 10 Issue 3 2018280
neutralize the free radical by accepting or donating an electron to eliminate the unpaired condition.[5-9] Antioxidant is effective in the treatment of cancer, coronary heart disease, and neurodegenerative diseases. It is also used in the treatment of Friedreich ataxia. Bioactive derivatives acting as flavonoids, stilbenoids, and alkaloids possess potent antioxidative and anti-inflammatory properties and are now being inducing in the treatment of Parkinsons disease. Flavonoids such as hesperidin have been proved to have an antioxidant effect against neurodegenerative disease. Structure-activity relationship of hesperidin to their antioxidant property showed that the effects of antioxidative activity of hesperidin depend on the number and order of OH groups and the presence of C4-C8 double bond conjugated pursuing 4-keto group in the flavonoid structure. In addition, it has potential activity in anti-inflammatory, antiviral, anticancer, sedative, Huntingtons stroke, and Alzheimer.
Apparently, the brain needs a regular supply of fuels, gases, and nutrients to maintain homeostasis and other vital function. The bloodbrain barrier represents an effective obstacle for the delivery of neuroactive agents to the central nervous system (CNS), and it makes the treatment of many CNS disease difficult to achieve. Activating A2A adenosine receptor (AR) temporarily augments the intracellular spaces between the brain capillary endothelial cells. A2A AR is a member of G protein-coupled receptor. Considering the properties of biodegradable nanoparticles essentially grand bioavailability, control release, less toxicity, and better encapsulation, they are frequently used as a vehicle in drug delivery. Apparently, a used biodegradable nanoparticle includes PLGA, PLA, chitosan, gelatin, polycaprolactone, and poly-alkyl-cyanoacrylates.[13-15] Chitosan is an amino polysaccharide which possesses the idiosyncratic features such as non-toxicity, hydrophilicity, biodegradability, mechanical strength. Biocompatibility and physical inertness are quotidianly used as a carrier for targeted drug delivery in neurodegenerative disorder. It possesses higher penetration capacity, absorption across the mucosal epithelia, and the potential of binding with various ligand molecules and helps in the formation of stable non-complex.[16-21] It is cationic in nature which involves in the formation of stable ionic complexes with multivalent anionic ions or polymers in various forms such as gels and micro/nanoparticles.[22-27] It has an antioxidant activity which is used here as a polymer produced from the exoskeleton of crustaceans (e.g., crabs and shrimps).[28-31] Hence, the present study is carried out to enhance the targeted drug delivery of hesperidin in combination with polymers for the treatment of Parkinsons disease.
MATERIALS AND METHODSHesperidin, chitosan, and tripolyphosphate (TPP) were purchased from Sigma Aldrich. All the other chemicals used were of analytical grade.
Incompatibility StudiesFourier-transform infrared (FTIR) studyFTIR analysis of pure hesperidin mixture (hesperidin, chitosan, and TPP) was performed, and the spectrum was obtained using FT-IR (Perkin Elmer 200 series). All spectra were recorded within a range of 2000650 cm1.
Differential Scanning Calorimetric Analysis (DSC)DSC measurements were carried out using NETZSCH DSC 204. Samples were analyzed in a temperature range of 0C300C at a heating rate of 5C/min under nitrogen atmosphere. The samples were prepared by pressing them in a DSC aluminum pans and subjected to analysis.
Preparation of Standard Plot of HesperidinAccurately weighed 100 mg of hesperidin dissolved in 100 ml standard volumetric flask using distilled water to get the stock solution of 100 mg/ml. From this stock solution, 10 ml of solution was withdrawn and made to 100 ml, and from these, aliquots of 1, 2, 3, 4, and 5 ml were withdrawn and further diluted to 10 ml with water to obtain a concentration range of 1050 g/ml. The absorbance of the solutions was measured at 263 nm using ultraviolet (UV)- spectrophotometer. A graph of concentration versus absorbance was plotted.
Preparation of Hesperidin Nanosuspension and Optimization of Preparation ProcessNanosuspension with relevant stabilizer was prepared by ionotropic gelation method. Chitosan solution (0.1%w/v) was prepared by dissolving chitosan in 100 ml of 1%v/v of acetic acid, and the resulting solution was stirred at 1500 rpm for 30 min on magnetic stirrer (Different concentrations of 0.1%, 0.2%, 0.3%, 0.4%, and 0.5% were prepared). TPP solution of 0.1% was prepared by dissolving 100 mg of TPP in 100 ml of deionized water. 100 mg of hesperidin was added to TPP solution and mix to form a homogenous mixture by stirring with a glass rod. Add the above mixture of TPP and hesperidin solution drop by drop (10 ml) to the chitosan solution and kept stirring at 2,500 rpm for 3 h on mechanical stirrer. Nanoparticles were obtained on the addition of a TPP and hesperidin solution to a chitosan solution. The nanoparticle suspension is then centrifuged at 15,000 rpm for 10 min using high-speed centrifuge (Sigma). Discard the sediment and preserve the supernatant. The formation of nanoparticles results in interaction between the negative groups of TPP and the positively charged amino groups of chitosan, as show