Formulation and evaluation of biodegaredable...
Transcript of Formulation and evaluation of biodegaredable...
Journal of Pharmacy Research Vol.11 Issue 2 February 2017
Prevesh Kumar et al. / Journal of Pharmacy Research 2017,11(2),96-102
96-102
Research Article
ISSN: 0974-6943
Available online throughhttp://jprsolutions.info
*Corresponding author.
Prevesh Kumar
Research Scholar,
OPJS Institute of Pharmacy,
OPJS University, Churu, Rajasthan. ,India
Formulation and evaluation of biodegaredable nanoparticleof Glimepiride by ionic gelation method
Prevesh Kumar1*, Dr. Devender Pathak1, Pawan Singh2
1*,2 Research Scholar, OPJS Institute of Pharmacy, OPJS University, Churu, Rajasthan. ,India.3Research Scholar, IFTM Institute of Pharmacy Academy, IFTM University, Moradabad, Uttar Pradesh, India.
Received on:24-12-2016; Revised on: 28-01-2017; Accepted on: 09-02-2017
ABSTRACT
Aim:To explore the water solubility of drug and increase oral bioavailability of Glimepiride. Nanoparticles of Glimepiride for oral drug
delivery were prepared by ionic gelation method using polyelectrolyte charge interaction. Glimepiride is an effective antidiabetic agent;
however, it suffers from short biological half-life. Therefore, it was selected as amodel drug. Method:The prepared Nanoparticles were
evaluated for Physico-chemical studies like drug polymer interaction through FT-IR analysis, Melting Point by Differential Scanning
Calorimetry, surface morphology by Scanning Electron Microscopy (SEM), Percent drug entrapment, production yield, in-vitro drug release
characteristics and release kinetics. The result of FT-IR studies showed that there was no drug polymer interaction found. Results and
Discussion:The SEM studies confirmed that with increase of polymer concentration the nanoparticles become smooth, and in-vitro release
studies showed that the drug release followed diffusion for formulation F5, F6, and F7 and for the formulation F8, F9 and F10 followed non-
fickian mechanism, moreover all the formulations exhibited high percentage yield as well as high percent drug entrapment. Conclusion:The
method proves to be beneficial in designing control release formulations of Glimepiride Chitosan-Gelatin-B ionic gelation method, using
polyelectrolyte charge interaction.
KEY WORDS: Nanoparticles, Glimepiride, Chitosan, Gelatin-B, ionic gelation method, Polyelectrolyte charge interaction.
1. INTRODUCTION:
Drug delivery is the method for administering a pharmaceutical
compound to achieve a therapeutic effect in humans or animals.
Drug delivery system can have very important role in theefficacy of
drugs [1]. Some drugs have an optimum concentration of range within
which maximum effect is derived. But there is very slow progress
inefficacy of the treatment of severe disease, has suggested a growing
need for drug delivery system [2]. Drug delivery system is amulti-
disciplinary approach to thedelivery of therapeutics to the target
tissue which gives new ideas on controlling the pharmacokinetics,
pharmacodynamics, immunogenicity, biorecognition, non-specific
toxicity, and efficacy of the drug [3].
The main approach of drug delivery system is to promoting the
exposure of drug on targeted area rather than anon-target area to
avoid unnecessary side effects.
Novel drug delivery system is based on two mechanisms,
1. Physical mechanism (osmosis, diffusion, erosion)
2. Biochemical mechanism (monoclonal antibiotics, gene
therapy, vector system)
Drug delivery system (DDS) such as biodegradable polymer based
nanoparticles can be designed to improve drug bioavailability orally.
There are many antibiotic, antifungal, anticancer drugs which are
improved by different drug delivery systems. DDS are designed to
alter the pharmacokinetics and biodistribution of the drug [4].
Polymeric Nanoparticles (PNP) are defined as aparticulate dispersion
or solid particles with a size range of 10 to 1000 nm in diameter [5].The
term PNP is a collective term given for any type of polymer
nanoparticles, but specifically for nanospheres and nanocapsules.
Nanospheres are matrix particles, i.e., particles whose entire mass is
solid and molecules may be adsorbed at the sphere surface or
encapsulated within the particle. In general, they are spherical, but
“nanospheres” with a nonspherical shape are also described in the
literature [6].
Journal of Pharmacy Research Vol.11 Issue 2 February 2017
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Nanocapsules are vesicular systems, acting as a kind of reservoir, in
which the entrapped substances are confined to a cavity consisting
of a liquid core (either oil or water) surrounded by a solid material
shell [7]. Nanoparticles may or may not exhibit size-related properties
that differ significantly from those observed in fine particles or bulk
materials.
The major goal in designing of polymeric nanoparticles as a delivery
system is,
1. To control particles size
2. Surface property
3. Release of pharmaceutical active agent to achieve in site
specification of at therapeutically optical range and dose
regimen
1.1. Ionotropic Gelation Method
Ionotropic gelation is based on the ability of polyelectrolytes to
cross-link in the presence of counter ions to form hydrogel beads
also called as gelispheres. Gelispheres are aspherical cross-linked
hydrophilic polymeric entity capable of extensive gelation and
swelling in simulated biological fluids and the release of drug through
it controlled by polymer relaxation[8].The hydrogel beads are
produced by dropping a drug-loaded polymeric solution into the
aqueous solution of polyvalent cations[9].The cationsdiffuse into
the drug-loaded polymeric drops, forming a three-dimensional lattice
of theionicallycrosslinked moiety. Biomolecules can also be loaded
into these gelispheres under mild conditions to retain their three-
dimensional structure[10].
Polyelectrolyte solution[Sodium Alginate (-)/Gellan gum (-)/CMC (-)/Pectin (-)/
Chitosan (+) + Drug]
Added drop wise under magnetic stirring by needle
Counter ion solution [Calcium chloride solution (+)/Sodium tripolyphosphate (-)]
Gelispheres
Glimepride (a BCS class II drug) is the third generation of asulfonyl
urea oral antidiabetic drug having high permeability and low
solubility. Low water soluble drugs often exhibit low dissolution
profile and oral bioavailability problems.
Glimepiride is a new low-dose oral sulfonylurea that provides 24-h
glycemic control of NIDDM with once-daily dosing. In experimental
animal models, glimepiride lowered blood glucose by stimulating
insulin release from the pancreas[11] also appeared to have extra
pancreatic effects (2-5). of note, glimepiride was associated with
fewer direct effects on the mammalian cardiovascular system than
other sulfonylureas. Glimepiride did not cause vasoconstriction in
an animal model of hypoxic lactic acidosis.[12] Glimepiride also induced
less alteration in coronary flow/resistance than glyburide. These
findings may be due to differences in relative effects on ATP-sensitive
K+ channels in cardiomyocytes and pancreatic 3-cells.
The pharmacokinetic profile in healthy subjects and NIDDM patients
suggests that this agent may be suitable for once-a-day dosing.
Glimepiride is rapidly and completely absorbed afteroral
administration, with subsequent biotransformation to metabolites
that lack clinically meaningful glucose-lowering activity.After multiple
doses, the apparent terminal half-life is 9 h (Hoechst-Roussel
Pharmaceuticals, unpublished observations).
These preclinical and clinical findings were the basis for the current
study, which was one in a series of dose-response studies.
The current study was designed to determine the preferred dosing
regimen and to evaluate the potential need for higher doses. Many
of NIDDM patients were randomized to receive glimepiride at 1) two
dosing intervals, once daily, or in two equally divided doses and
at 2) two concentration dosages, 8 and 16 mg daily.
2. MATERIALS AND METHODS
Glimepiride was used and provided by Bal Pharma Pvt Ltd, Ruderpur,
India., Chitosan, and Gelatin-B was obtained as gift sample from Hi-
Media laboratories Pvt. Ltd Mumbai. Sodium Tri-poly phosphate
was obtained as gift sample from Sigma-Aldrich Pvt Ltd Mumbai.
Light liquid paraffin and Heavy liquid paraffin was obtained as gift
sample from CDH Pvt, Ltd, New Delhi.
2.1. Method
The Glimepiride nanoparticles were prepared by ion gelation
technique by using chitosan/gelatin-B mixture as a coating material.
Chitosan and gelatin-B were dissolved in dilute acetic acid solution
(1% v/v) together at concentrations of 1–4% w/v and adjusted to a
certain solution pH (usually 5.0). Glimepiride (10mg) was dissolved
in the above polymeric mixture. The drug in the polymeric mixture
was emulsified in 200 ml of liquid paraffin (1:1 mixture of light and
heavy liquid paraffin) at 40°C containing 1 ml Tween 80 (2% w/v).
The emulsification time was allowed for 10min under mechanical
stirring (500 rpm). The w/o emulsion was cooled to 4°C to induce
coagulation of Gelatin-B. Then 50 ml Na-TPP (1% w/v) with pH in the
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range 4–5 at 4°C was added dropwise. Stirring was continued for 15–
60 min to obtain cross-linked nanoparticles. Nanoparticles were
collected by centrifugation and washed with double distilled water
several times, then with acetone to remove water and dried at room
temperature under vacuum. The prepared nanoparticles were stored
in desiccators for further studies. Glimepiride loaded nanoparticles
with different polymer compositions (1:5, 1:6, 1:7, 1:8, 1:9 and 1:10)
were named as F5, F6, F7, F8, F9, and F10, respectively.
2.2. EVALUATION PARAMETERS
2.2.1. Percentage yield determination
2.2.1.1. Percentage Drug Entrapment (PDE)
2.2.1.2. Particle size analysis
Particle size was determined by optical microscopy. Briefly, about 5
mg of nanoparticles were taken on a glass slide and sizes of about
200 spherical particles were measured each time (n=3) by using an
optical microscope.
2.2.1.3.Surface morphology and dimensional analysis
The morphological and dimensional analyses of the nanoparticles
were performed by scanning electron microscopy (SEM).
Nanoparticles size and size distribution were determined by SEM
photomicrographs analyzing about 500 nanoparticles.
2.2.1.4. In vitro drug release
Drug release studies were carried out using USP XXIII basket
dissolution rate test apparatus (100 rpm, 37 ± 1°C) for 2h in 0.1 N HCl
(with 0.5% SLS) pH=1.2 and for 8h in 7.4 pH phosphate buffer (with
0.5% SLS).
At different time intervals, 5ml of the sample was withdrawn and
replaced with the same amount of fresh medium. The sample was
analyzed for Glimepiride directly or after appropriate dilution with
the pH 7.4 phosphate buffer spectrophotometrically at 233 nm using
a UV/ VIS spectrometer against a reagent blank.
3. RESULT AND DISCUSSION
3.1. Drug and polymer compatibility study
3.1.1. FTIR Study of Drug and chitosan
The FTIR spectra of the physical mixture (1:1) of Drug and Chitosan
showed slight shifting in absorption bands, but the characteristic
group for –NH stretching were found to be an intact i.e. peak at
3333.69 cm-1 and 3288.77 cm-1, moreover the peak at 1674 cm-1was
also found to be intact.
Fig 1. FTIR Study of Drug and Chitosan
3.1.2. Drug and Gelatin-BThe FTIR spectra of a physical mixture of Glimepiride and Gelatin-B(1:1) showed no interaction with the drug as major peaks of the drugfound to be intact in the spectra. The characteristic peak for –NHstretching were found to be an intact i.e. peak at 3333.69 cm-1 and3288.77 cm-1, moreover the peak at 1674 cm-1 was also found to beintact.
Fig 2. FTIR Study of Drug and Gelatin-B.
3.1.3. Differential Scanning Calorimetry (DSC):
The thermal behavior of drugs and excipients was studied by a TA
Instruments DSC Q20. The scans were carried out on each sample, at
scan rates of 10 °C/min
This fig show the pure drug melting point showing the
purity of drug and the melting point was found is 207 ºC
Fig 3. DSC thermogram of Glimepiride
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This fig show the Gelatin-B melting point showing Gelatin-B the melting point was found is less than 35 ºC
Fig 3. DSC thermogram of Glimepiride
This fig show the Gelatin-B melting point showing Gelatin-B the melting point was found is less than 35 ºC
Fig 4. DSC thermogram of Gelatin- B
This figure shows the Sodium tripolyphosphate melting pointshowing Sodium tri-polyphosphate the melting point was found is522 ºC.
This fig show the Chitosan melting point showing Chitosan themelting point was found is 203 ºC
Fig 5. DSC thermogram of Sodium tri-polyphosphate
Fig 6. DSC thermogram of Chitosan
Fig 7.DSC thermogram of Drug-loaded
S. No Formulation code Production yield (%)
1 F5 8 5
2 F6 6 9
3 F7 9 3
4 F8 5 6
5 F9 6 0
6 F10 6 1
In this fig showing the no drug interaction with the nanoparticlesand the pure drug melting point is shifted from the original meltingpoint that showing the entrapment of drug in nanoparticles.
3.1.4. Percentage yield determination
The production yield of chitosan-Gelatin-B nanoparticle prepared
with thedifferent concentration of polymer at 500 rpm is shown in
thetable. The production yield was found to decrease with increase
in the composition ratio of drug and polymer, the decrease in the
percentage yield could be attributed due to the higher cross-linking
between oppositely charged polyelectrolyte.
Table 1:Percentage yield of Different Formulation of PreparedNanoparticles
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Fig 8. Percentage Yield of different Formulations
S. No Formulation code Entrapment efficiency(% )
1 F5 9 62 F6 9 03 F7 9 24 F8 9 65 F9 9 56 F10 9 4
Fig. 9 Percentage Drug Entrapment of different Formulations
3.1.6. Particle Size AnalysisThe main particle size of different nanoparticles was found to be82.02 to 126.8 nm, table. The increase in the gelatin-B concentrationaffected the particle size of nanoparticles, at aconcentration (1:5) themean particle size was found to be 85.03 nm, whereas at aconcentration(1:7) it was 126.8 nm. This increase in particle size may be due toincrease the viscosity of droplets with anincrease in polymerconcentration, which resulted in larger droplets of theemulsion. Table3 represents the data.
Fig 10. Particle size of different formulations analyzed by TEM
Fig 11. Particle size of different Formulations analyzed by Zeta
Sizer
Table 3 Polydispersity index (PDI) and ZETA SIZER
S. No Formulation code Mean partic le size(nm)
1 F5 85.32 F6 89.83 F7 126.84 F8 82.025 F9 96.756 F10 88.32
3.1.5.Percentage Drug EntrapmentThe entrapment efficiency of all the prepared formulations compliedwith the Pharmacopoeial limits for the drug content. The entrapmentefficiency of the drug in the prepared nanoparticles was found to bebetween 90 to 96%, table.
Table 2 Percentage Drug Entrapment Efficiency of DifferentFormulations
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Fig 12. Particle size of different Formulations
Fig 13. Cumulative % Drug Release of Glimepiride fromChitosan- Gelatin-B Nanoparticles
Table 4.Cumulative % Drug Release of Glimepiride from Chitosan- Gelatin B Nanoparticle
3.1.8.In vitro drug release
3.1.8.1..Influence of change in pH of dissolution medium
3.1.7.Polydispersity index (PDI)
PDI value range from 0 to 1, a higher value indicates a less
homogenous nanoparticles size distribution. Following figure indicate
that PDI is almost 0.268 which means formulations had a homogenous
distribution of nanoparticles in water shown in above fig.
The prepared polyelectrolyte exhibited drug release in a controlled
manner at pH 7.4 and it was also found that the drug release was
decreased as the concentration of polymer was increased. The drug
release rate could be slower because of the diffusion of Glimepiride
from the internal nanoparticles environment. This indicates that cross-
linked Chitosan-Gelatin-B nanoparticles should have produced
stronger electrostatic interaction with negatively charged Na-TPP
(Polyanion).
Time (hrs) Cumulative % Drug Release
F5 F6 F7 F8 F9 F10
0 0 0 0 0 0 0
1 37.39 ± 1.2 30.09 ± 1.10 26.11 ± 0.34 16.33 ± 1.22 12.1 ± 2.45 10.23 ± 3.44
2 42.22 ± 0.68 35.1 ± 1.19 29.67 ± 0.23 20.89 ±1.67 18.78 ± 1.28 14.47 ± 1.22
3 45.39 ± 1.7 38.22 ±1.20 34.32 ± 1.22 28.38 ± 1.00 25.28 ± 2.22 19.68 ± 1.10
4 49.12 ± 2.3 42.89 ± 2.33 39.19 ± 2.22 32.87 ± 1.27 30.23 ± 1.25 25.47 ± 2.69
5 54.34 ± 1.9 46.12 ± 2.63 44.28 ± 2.19 37.12 ± 0.78 34.47 ± 1.27 30.48 ± 2.37
6 60.59 ± 2.0 50.28 ± 3.09 47.66 ± 1.99 41.86 ± 0.34 39.89 ± 0.24 35.49 ± 2.00
7 68.89 ± 1.45 55.48 ± 3.67 52.34 ± 1.23 47.28 ± 2.67 45.37 ± 1.00 40.88 ± 3.45
8 73.34 ± 1.00 59.67 ± 1.23 56.23 ± 0.10 53.81 ± 1.23 49.67 ± 2.89 45.38 ± 2.36
9 79.32 ±1.21 65.23 ± 0.45 60.48 ± 0.23 57.48 ± 1.00 54.34 ± 2.67 49.48 ± 3.33
1 0 84.27 ± 1.23 70.11 ± 1.22 64.79 ± 0.34 61.39 ± 2.34 58.23 ± 2.34 55.39 ± 3.25
11 90.45 ± 1.26 75.1 ± 1.65 68.67 ± 0.12 65.32 ± 1.45 63.01 ± 1.35 60.02 ± 1.23
1 2 97.99 ± 1.28 79.09 ± 1.11 75.56 ± 1.23 71.78 ± 1.34 68.89 ± 1.26 66.12 ± 2.25
Table 5. Mechanism of drug release
Formulation Code Zero Order First Order Higuchi Model KorsmeyerPeppas n Value
F5 0.922 0.800 0.992 0.934 0.254
F6 0.917 0.963 0.978 0.958 0.325
F7 0.933 0.970 0.985 0.963 0.397
F8 0.977 0.976 0.974 0.986 0.596
F9 0.985 0.989 0.972 0.997 0.696
F10 0.996 0.983 0.948 0.988 0.772
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4. CONCLUSIONThe process was found to have great potential in producingnanoparticles of uniform size, with asmooth surface and very highpercent drug entrapment efficiency coupled with high productionyield. It was also found that polyelectrolyte complex formationbetween the positively charged Chitosan and negatively chargedGelatin B can control the release of model drug i.e. Glimepiride forabout 12 hours in almost all the formulations, as this system can beuseful in designing control release formulation through simplemodifications in formulation parameters. The most importantparameter that was found to affect were polyelectrolyte concentration,pH of the formulation medium, stirring rate, thecrystallinity of thedrug.
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Source of support: Nil, Conflict of interest: None Declared