Development and Characterization Pioglitazone Hydrochloride Loaded Nanoparticle for Management of Diabetes Mellitus
Department of Pharmacy, Guru Ghasidas Vishwavidyalaya, Bilaspur – 495009, C.G. India.
*Corresponding Author E-mail: meenapharmaceutics@rediffmail.com
ABSTRACT:
The potential applications of nanoparticles as oral drug delivery systems for diabetes treatment are discussed in this research article. This paper discusses polymeric nanoparticles, polysaccharides, and polymeric nanoparticles in the oral administration of anti-diabetic drugs. Diabetes is a chronic metabolic disorder characterized by insulin deficiency. Diabetes treatments are numerous, and nanoparticles have several advantages. Several studies and research reports based on nanotechnological approaches in the formulation of anti-diabetic drugs have highlighted the fact that research in the formulation of nanodrugs improved diabetes-fighting strategies based on plausible molecular mechanisms of action of the drugs. Additionally, efforts have been made to determine the optimal drug concentration and time of exposure in order to recommend a scientifically validated drug dose response in the development of various therapeutic strategies for anti-diabetic drugs, various types of nanoparticles are available; polymeric nanoparticles are one of the most commonly used nanoparticles. Polymeric nanoparticles range in size from 10-1000 nm. Polymeric nanoparticles created by combining a drug and a polymer. The main advantages of polymeric nanoparticles are their ease of preparation, targeted delivery, dose minimization, and high therapeutic efficiency.
KEYWORDS: anti-diabetic, hypoglycemic drug, polymeric nanoparticles, polymers, Pioglitazone, oral delivery, Solvent evaporation method.
INTRODUCTION:
Diabetes mellitus is an endocrine disease in which the pancreas does not produce sufficient insulin or the body cannot efficiently use the insulin it produces 1,2. Diabetes mellitus (DM), commonly known as diabetes, is a group of metabolic disorders characterized by hyperglycemia (High blood sugar) resulting from defects in insulin secretion, insulin action, or both3. Diabetes mellitus (DM) is probably one of the oldest diseases known to man. It was first reported in Egyptian manuscript about 3000 years ago 4.
In 1936, the distinction between type 1 and type 2 DM was clearly made. Type 2 DM was first described as a component of metabolic syndrome in 1988. Type 2 DM (formerly known as non-insulin dependent DM) is the most common form of DM characterized by hyperglycemia, insulin resistance, and relative insulin deficiency5. Type 2 DM results from interaction between genetic, environmental and behavioral risk factors6,7. Polymer-based NP (PNP) is a collective term which is given to any kind of polymer NP, but specifically is applied for nanospheres and nanocapsules. Polymeric nanoparticles (PNPs) have gained consideration in numerous research activities and have been deployed in encouraging number of domains during the last few years. They have been speculated as nanosized solid colloidal particles engendered from polymers. Nanoparticles originating from polymers are generally inter-connected with novel properties. Eminent factors causing PNPs to attain relatively unique properties than bulk masses are connected with their reduced particle size8. Polymeric nanoparticles are commonly 10-1000nm in dimension. These polymeric nanoparticles are formulated from polymers, which have the nature of bio-adaptability, bio-compatibility, and bio-degradable. The drug is dissolved, entrapped, and encapsulated to a nanoparticle, nano-spheres or nano- capsule is obtained depending upon the preparation. In a nano-capsule system, the drug is limited to a cavity enclosed by an even polymer layer, while the nano-shell contains of a medium, in which the drug is physically and uniformly dispersed9.
MATERIAL AND METHOD:
The Pioglitazone Hydrochloride was gift sample from Sun Pharmaceutical Industries Ltd., Gurgaon, Delhi, India.
Solvent Evaporation Method for preparation of nanoparticles:
Solvent evaporation method first developed for preparation of nanoparticles 10. This method involves two steps. The first step is emulsification of polymer in aqueous phase and in second step is evaporation of polymer solvent 11. In this method firstly nanoemulsion formulation prepared. Polymer dissolved in organic solvent (Dichloromethane, Chloroform or ethyl acetate). Drug is dispersed in this solution. Then this mixture emulsified in an aqueous phase containing surfactant (Polysorbates, poloxamers sodium dodecyl sulfates polyvinyl alcohol, gelatin) make an oil in water emulsion by using mechanical stirring, sonification, or micro fluidization (high pressure homogenization through narrow channels). After the formation of emulsion, the organic solvent evaporates by increased the temperature and reduced pressure with continuous stirring12.
The natural types of polymers are the obtained from the natural sources like animal and plants. The synthetic polymers are the prepared by the chemical synthesis method 13. The types of polymers are can be used for the polymeric nanoparticle’s formulations are shown in figure.2 Pioglitazone Hydrochloride loaded eudragit nanoparticles prepared by solvent evaporation method. Different batches of nanoparticles F1, F2, F3, and F4 were prepared by using different concentration of polymer. The drug (Pioglitazone Hydrochloride) was dissolved in 10 ml of dichloromethane and acetone (each 5ml) and polymer (Eudragit RS100) in the ratio 1:1, 1:2, 1:3 and 1:4 was dissolved in 10ml of dichloromethane and then added drop wise to the 100ml of aqueous phase containing 0.1% w/v of PVA as the emulsifying agent. The emulsion 14 was stirred at the stirring speed of 1000rpm using Euro Star high speed stirrer (IKA Labortechnik, Germany) for 4h under room temperature to evaporate the organic phase. The milky emulsion centrifuged at 10000 rpm for 30 min. Then nanoparticle was lyophilized (-500C for 24 h) and used further characterization15.
Surface charge properties of the nanoparticle are studied through zeta potential. The value of particle surface charge indicates the stability of nano- suspension at the microscopic level. Zeta potential of Pioglitazone Hydrochloride loaded nanoparticles a Malvern Zetasizer (Nano-ZS, UK). The value of particle surface charge indicates the stability of nanoparticles at the macroscopic level. Samples were diluted (50 folds) using distilled water and then analysis was performed at 25 ºC and 149 watts. Each sample of the nanoparticle was adjusted to a concentration of 0.05% (w/v) in filtered water in the case of zeta potential examination. The zeta potential was determined in triplicate for a single batch of nanoparticle and the result was recorded as the average of three measurements16. Were shown in fig.1.
Table 1: Formulation code
|
S.No. |
Formulation code |
Polymer ratio (%w/v) |
Time (hrs.) |
Stirring speed (rpm) |
|
1 |
F1 |
1:1 |
4 |
1000 |
|
2 |
F2 |
1:2 |
4 |
1000 |
|
3 |
F3 |
1:3 |
4 |
1000 |
|
4 |
F4 |
1:4 |
4 |
1000 |
Figure 1: Particle size of different formulation of Pioglitazone nanoparticle
Table 2: Zeta potential of different formulation
|
S.No. |
Formulation code |
Zeta potential (Mv) |
|
1 |
F1 |
3.88± 0.31 |
|
2 |
F2 |
3.63± 0.58 |
|
3 |
F3 |
35.37± 7.5 |
|
4 |
F4 |
9.74± 2.04 |
values are presented as mean±SD, n=3
Figure 2: Percentage encapsulation of Pioglitazone nanoparticles
TEM was used in the morphological analysis of Pioglitazone nanoparticles. The sample was prepared by casting one drop of freshly made nano- suspension onto the copper grid support and the extra solution was removed by filter paper. The external and internal morphology of the nanoparticle were studied using transmission electron microscopy (TEM). The Morgagni 268D, Fei USA Electron Optic were used for analysis of morphology of pioglitazone nanoparticle 17. Were shown in fig.3.
Figure 3: Transmission electron microscopy (TEM) of nanoparticles.
Differential scanning calorimetry of pioglitazone, Mβ-CD, physical mixtures and inclusion complexes were conducted using Differential scanning calorimetry (DSC) Q2000 V24.2 Build 107 instrument. The mass of empty pan and reference pan were taken into account for calculation of heat flow. The sample mass varied from 3-10 ± 0.5 mg and it was placed in sealed aluminum pans. The coolant used was liquid nitrogen. The samples were scanned at 10°/min from 20° to 300°. The temperatures of the characteristic transitions of formulation (F1), onset (T0) 153.49oC, peak (TP) 176.65 oC, and conclusion (TC) 182.59 oC and Eudragit RS 100 Polymer, onset (T0) 22.70 oC, peak (TP) 69.30 oC, and conclusion (TC) 86.28 oC, were recorded. The DSC thermo grams of plain EudragitRS100 and formulation of nanroparticle were recorded. The DSC of sample was performed in SOS, Department of Physics, Pt. Ravishankar Shukla University Raipur, India. Were shown in fig. 5-6.
Figure 4: Zeta potential of formulation F1
The entrapment efficiency of drugs (i.e., Pioglitazone) in Eudragit RS100 nanoparticles was assessed in directly determining the drug (unentrapped) using UV visible spectrophotometric method by applying the following equation:
Total amount of drug – Free drug
Drug entrapment (%) = ----------------------------- x 100
Total amount of drug
Free drug was removed by centrifugation using ultracentrifuge at 3000 rpm for 12min (Remi Cooling ultracentrifuge, Mumbai, India) and washed twice. Supernatant was collected and analyzed by UV Spectrophotometer (Shimadzu 1800, Japan) at 268 nm for Pioglitazone. The percentage drug entrapped and % yield was calculated as well as graphically presented in Figure.2.
In-vitro release studies were performed using the membrane diffusion technique 20-26. The membrane was soaked before use in distilled water for 4 hours then rinsed thoroughly in distilled water. 1 ml of pioglitazone hydrochloride nanoparticles dispersions, equivalent to (1.5mg/ml) of pioglitazone was transferred in to dialysis membrane bag, tied and placed in beaker containing 100ml of Dissolution medium. The entire system was kept at 37°±0.5° with continuous magnetic stirring (50rpm) and the study was carried out in two dissolution media; 0.1N HCl pH 1.2. At appropriate time intervals 5ml of release medium was removed and 5ml fresh medium was added into the system to maintain sink condition. The amount of pioglitazone in the release medium was evaluated by U.V Spectrophotometer at 268nm.
Table 3: Release kinetic of different formulation
|
Correlation coefficient (R2) |
|||||
|
Formulations |
Zero order |
First order |
Higuchi model |
Hixon crowel Model |
Peppas model |
|
F1 |
0.807 |
0.872 |
0.952 |
0.851 |
0.672 |
|
F2 |
0.770 |
0.834 |
0.938 |
0.813 |
0.633 |
|
F3 |
0.798 |
0.864 |
0.954 |
0.843 |
0.649 |
|
F4 |
0.824 |
0.886 |
0.964 |
0.866 |
0.700 |
Figure 5: DSC Thermogram of Eudragit RS 100
Figure 6: DSC Thermogram of Formulation
Figure 7: Percentage cumulative drug release from different formulation
Cumulative % drug release v/s. time (zero order kinetic models), Log cumulative % drug retain v/s. time (first order kinetic model), Cumulative % drug release v/s. square root of time (Higuchi kinetic model)., Log cumulative % drug release v/s. log time (Korse Meyer-Peppas kinetic model.), Cube roots of cumulative % drug retain v/s. time (Hixson-Crowell kinetic model). Were shown in table 3.
WHO states that, the stability of finished pharmaceutical products depends on environment factors such as ambient temperature, humidity and light as well as on the product related factors e.g chemical and physical properties of active substance and of pharmaceutical excipient, the dosage form and its composition, the manufacturing process, the nature of the container closure system and properties of packaging material 27.
Nanoparticle 28-31 was kept at a temperature of 25 ±2oC and at 40 ±2oC for a two month and then the in- vitro drug release of optimized formulation (F1) was performed. Were shown in fig.8.
Figure 8: Effect of temperature on in-vitro drug release of formulation
The solvent evaporation technique was used to create pioglitazone hydrochloride loaded nanoparticles. The use of factorial design resulted in a statistically systematic approach for the formulation of nanoparticles with the desired particle size, high entrapment efficiency, and % drug release of pioglitazone hydrochloride loaded eudragit RS 100 nanoparticles. The optimized formulation (F1) released 68.90% of the drug in vitro in 8 hours.
No conflict of interest.
REFERENCES:
1. Sonia TA, Sharma CP. An overview of natural polymers for oral insulin delivery. Drug Discov Today. 2012; 17:784-92.
2. Ramesan RM, Sharma CP. Challenges and advances in nanoparticle-based oral insulin delivery. Expert Rev Med Devic. 2009; 6:665-76.
3. World Health Organization. About diabetes, Archived from the original on 31 March 2014. Retrieved 4 April 2014.
4. Ahmed AM. History of diabetes mellitus. Saudi Med J. 2002; 23(4): 373-378.
5. Simos YV, Spyrou K, Patila M, Karouta N, Stamatis H, Gournis D, Dounousi E, Peschos D. Trends of nanotechnology in type 2 diabetes mellitus treatment. Asian Journal of Pharmaceutical Sciences. 2020; https://doi.org/10.1016/j.ajps.2020.05.001
6. Maitra A, Abbas AK. Endocrine system. In: Kumar V, Fausto N, Abbas AK (eds). Robbins and Cotran Pathologic basis of disease. Philadelphia, Saunders 2005;7:1156-1226.
7. Chen L, Magliano DJ, Zimmet PZ. The worldwide epidemiology of type 2 diabetes mellitus: present and future perspectives. Nature Reviews Endocrinology. 2011.
8. Okur ME, Karantas ID, Siafaka PI. Diabetes Mellitus: A Review on Pathophysiology, Current Status of Oral Medications and Future Perspectives. Acta Pharmaceutica Sciencia. 2017; 55: 60-82. doi: 10.23893/1307-2080.APS.0555
9. Rao JP, Geckeler KE. Polymer nanoparticles: Preparation techniques and size-control parameters. Progress in Polymer Science. 2011; 36:887–913.
10. Francis R, Joy N, Aparna EP, Vijayan R. Polymer grafted inorganic nanoparticles, preparation, properties, and applications: A review. Polym. Rev. 2014;54: 268–347.
11. Ranjit K, Baquee AA. Nanoparticle: A review of preparation and characterization and application. IRJP. 2013; 4:48-52.
12. Nagavarma BVN, Yadav HKS, Ayuz A, Vasudha L S, Shivakumar HG. Different technique for preparation of polymeric nanoparticles – A Review, Asian Journal of Pharmaceutical and Clinical Research. 2012;5:3:8: 1-8.
13. Todaro B, Moscardini A, Luin S. Pioglitazone-Loaded PLGA Nanoparticles: Towards the Most Reliable Synthesis Method. International Journal of Molecular Sciences. 2022; 23:2522. https://doi.org/ 10.3390/ijms23052522
14. Mohanraj VJ, Chen Y. Nanoparticles- a review. Tropical Journal of Pharmaceutical. 2006; 5:1:561-573.
15. Canchi A, Khosa A, Singhvi G, Banerjee S, Dubey SK. Design and Characterization of Polymeric Nanoparticles of Pioglitazone Hydrochloride and Study the Effect of Formulation Variables using QbD Approach. Current Nanomaterials. 2017; 2: 162-168.
DOI: 10.2174/2405461503666180501115359
16. Molepeeceres J, Guzman M, Abruturas M R, Chacon M, Berger L. Application of center composite design in preparation of Polycaprolactone Nanoparticles by solvent displacement method. Int J Pharm Sci. 1996; 85:2006-213.
17. Meyer E, Heinzelmann H. Scannin force microscopy. In: Wiesendanger R, Guntherodt H J. Editors. Scanning tunneling microscopy II, Surface Science. New York: Springer Verlag 1992;99-149.
18. Khachane KN, Bankar VH, Gaikwad PD, Pawar SP. Formulation, evaluation and optimization of sustained release drug delivery of trimetazidine dihydrochloride by full factorial design. Inventi Impact: Pharm Tech. 2012; 3: 205-211.
19. Ghorab MM, Tag R, Salah S. Preparation and characterization of ofloxacin loaded biodegradable poly D, L-lactide nanoparticles: A factorial design study. Inventi Impact: Pharm Tech. 2012; 2: 121-128.
20. Mullaicharam A R, Nanoparticle in drug deliver y system, International Journal of Nutrition, Pharmacology Neurological Disease. 2011;1:2 103-121.
21. Cetin M, Atila A, Sahin S, Vural I. Preparation and characterization of metformin hydrochloride loaded-Eudragit RSPO and Eudragit RSPO/PLGA nanoparticles. Pharmaceutical Development and Technology, 2013; 18(3): 570–576. doi: 10.3109/10837450.2011.604783
22. Muzib YI, Ramya E, Ambedkar YR. Formulation, Optimization and Pharmacodynamic Studies of Pioglitazone HCl Solid Lipid Nanoparticles. Journal of Pharmaceutical Research International. 2021; 33(46A): 329-341.doi: 10.9734/JPRI/2021/v33i46A32873
23. Rojewska A, Karewicz A, Karnas K, Wolski K, Zajac M, Kaminski K, Szczubiałka K, Zapotoczny S, Nowakowska M. Pioglitazone- Loaded Nanostructured Hybrid Material for Skin Ulcer Treatment. Materials. 2020; 13: 2050; doi:10.3390/ma13092050
24. Boddupalli BA, Masana P, Anisetti RN, Kallem SV, Madipoju B. Formulation and evaluation of Pioglitazone loaded Bovine serum albumin nanoparticles along with Piperine. Drug Invention Today. 2013; 5:212-215. http://dx.doi.org/10.1016/j.dit.2013.05.011
25. Souto EB, Souto SB, Campos JR, Severino P, Pashirova TN, Zakharova LY, Silva AM, Durazzo A, Lucarini M, Izzo AA, Santini A. Nanoparticle Delivery Systems in the Treatment of Diabetes Complications. Molecules. 2019; 24:4209 doi:10.3390/molecules24234209
26. Emin Cam M, Ertas B, Alenezi H, et al. Accelerated diabetic wound healing by topical application of combination oral antidiabetic agents- loaded nanofibrous scaffolds: An in vitro and in vivo evaluation study. Materials Science and Engineering. 2020; https://doi.org/10.1016/j.msec.2020.111586
27. Tangri P, Bisht B. Who Role and Guidelines in Stability Study of Pharmaceuticals: A Regulatory Perspective. International Journal of Research in Pharmaceutical and Biomedical Science 2012;3(3).
28. Kamela R, batanonyb RE, Salama A. Pioglitazone-loaded three-dimensional composite polymeric scaffolds: A proof of concept study in wounded diabetic rats. International Journal of Pharmaceutics. 2019; 570 :118667 https://doi.org/10.1016/j.ijpharm.2019.118667
29. Ding Y, Cheng Y, Hao B, Zhu L, Zhang N, Zhao B, Tian Y. Metal–organic framework modifed by silver nanoparticles for SERS based determination of sildenafil and pioglitazone hydrochloride. Microchim Acta. 2021;188: 351 ttps://doi.org/10.1007/s00604-021-05008-4
30. Shehata TM, Almostafa MM, Elsewedy HS. Development and Optimization of Nigella sativa Nanoemulsion Loaded with Pioglitazone for Hypoglycemic Effect. Polymers. 2022;14: 3021. https://doi.org/10.3390/polym14153021
31. Shalu Choudhary , unil. The Effect of Temperature and Pressure Dependent Viscosity on Thermal Convection in a Rotating Couple-Stress Fluid Saturating a Porous Medium: A Nonlinear Stability Analysis. Research J. Science and Tech. 2013; 5(1): 55-64.
Received on 31.03.2023 Modified on 01.06.2023
Accepted on 04.07.2023 © RJPT All right reserved
Research J. Pharm. and Tech 2024; 17(4):1748-1752.
DOI: 10.52711/0974-360X.2024.00277