• SRINIVAS MURTHY BR Research Scholar, Department of Pharmaceutical Sciences, Jawaharlal Nehru Technological University Anantapur, Anantapuramu, Andhra Pradesh, India.
  • PRASANNA RAJU YELAVARTHI Department of Pharmaceutics Division, Sri Padmavathi School of Pharmacy, Tiruchanoor, Tirupati, Andhra Pradesh, India.
  • DEVANNA N Department of Chemistry and Director, Oil Technological and Pharmaceutical Research Institute, Jawaharlal Nehru Technological University Anantapur, Anantapuramu, Andhra Pradesh, India.


Objective: High lipophilicity and extensive hepatic metabolism limit oral application of orlistat in obesity treatment. Orlistat-loaded chitosan nanoparticles (CONPs) were optimized by 3-factor 3-level Box–Behnken design (BBD) and surfaced engineered to address limitations.

Methods: CONPs were prepared by ionic gelation method. Amounts of chitosan (X1), sodium tripoly phosphate (X2), and orlistat (X3) were selected as independent factors, whereas % entrapment efficiency (Y1) and % drug release (Y2) were employed as responses in BBD. Three-dimensional response surface plots were run to understand the main interaction and quadratic effects of independent variables. Further optimized formulation was surface engineered by Eudragit L-100 (ECONPs) and characterized by FTIR, DSC, XRD, particle size, zeta potential, and SEM. Entrapment efficiency, release kinetics, stability, and in vitro cell line studies were carried out.

Results: ECONPs were produced with an average size of 534.6 nm, zeta potential of +5.7 mV, EE of 78.62%, and DR of 80.86%. Eudragit coated CONPs anchored the release of orlistat at pH 6.8 desirable for duodenal targeting. Orlistat was released with low, burst, and sustained release manner over 24 h period followed first-order kinetics with Higuchi model with drug content of 84.87% and 78.44% of release. ECONPs possessed lipase inhibition with IC50 value of 8.0 μg/ml and viability against selected cell lines with CTC50 values (26.32–32.21 μg/ml).

Conclusion: BBD was a promising tool in elucidating the insights of formulation variables of CONPs. ECONPs fulfilled the rationale of orlistat release, lipase inhibition, and viability against selected cell lines.

Keywords: Chitosan, Hyperlipidemia, Ionic gelation, Orlistat, Response surface method


1. Yumuk V, Tsigos C, Fried M, Schindler K, Busetto L, Obesity Management Task Force of the European Association for the Study of Obesity. European guidelines for obesity management in adults. Obes Facts 2015;8:402-24.
2. Blackburn GL, Wollner S, Heymsfield SB. Lifestyle interventions for the treatment of class III obesity: A primary target for nutrition medicine in the obesity epidemic. Am J Clin Nutr 2010;91:289-92.
3. Finer N. Medical consequences of obesity. Medicine 2015;43:88-93.
4. Sabrin RI, Gamal AM, Zainy MB. Natural antihyperlipidemic agents: Current status and future perspectives. Phytopharmacology 2013;4:492- 531.
5. Henness S, Perry CM. Orlistat: A review of its use in the management of obesity. Drugs 2006;66:1625-56.
6. Mittendorfer B, Ostlund RE, Patterson BW, Klein S. Orlistat inhibits dietary cholesterol absorption. Obes Res 2001;9:599-604.
7. Hill TK, Davis AL, Wheeler FB, Kelkar SS, Freund EC. Development of a self-assembled nanoparticle formulation of orlistat, Nano-ORL, with increased cytotoxicity against human tumor cell lines. Mol Pharm 2016;13:720-8.
8. Sangwai M, Sardar S, Vavia P. Nanoemulsified orlistat-embedded multi-unit pellet system (MUPS) with improved dissolution and pancreatic lipase inhibition. Pharm Dev Technol 2014;19:31-41.
9. Dolenc A, Govedarica B, Dreu R, Kocbek P, Srcic S, Kristl J. Nanosized particles of orlistat with enhanced in vitro dissolution rate and lipase inhibition. Int J Pharm 2010;396:149-55.
10. Rukmangathen R, Yallamalli IM, Yalavarthi PR. Biopharmaceutical potential of selegiline loaded chitosan nanoparticles in the management of Parkinson’s disease. Curr Drug Discov Technol 2019;16:417-25.
11. Jabr-Milane L, Van Vlerken L, Devalapally H, Shenoy D, Komareddy S. Multi-functional nanocarriers for targeted delivery of drugs and genes. J Control Release 2008;130:121-8.
12. Naghibi Beidokhti HR, Ghaffarzadegan R, Mirzakhanlouei S, Ghazizadeh L, Dorkoosh FA. Preparation, characterization, and optimization of folic acid-chitosan-methotrexate core-shell nanoparticles by Box-Behnken design for tumor-targeted drug delivery. AAPS PharmSciTech 2017;18:115-29.
13. Nguyen N, John JB. New 3-level response surface designs constructed from incomplete block designs. J Stat Plan Inf 2008;138:294-305.
14. Li P, Yang Z, Wang Y, Peng Z, Li S, Kong L, et al. Microencapsulation of coupled folate and chitosan nanoparticles for targeted delivery of combination drugs to colon. J Microencapsul 2015;32:40-5.
15. Rawal T, Mishra N, Jha A, Bhatt A, Tyagi RK, Panchal S, et al. Chitosan nanoparticles of gamma-oryzanol: Formulation, optimization, and in vivo evaluation of anti-hyperlipidemic activity. AAPS PharmSciTech 2018;19:1894-907.
16. Gajra B, Patel RR, Dalwadi C. Formulation, optimization and characterization of cationic polymeric nanoparticles of mast cell stabilizing agent using the Box-Behnken experimental design. Drug Dev Ind Pharm 2016;42:747-57.
17. Honary S, Ebrahimi P, Hadianamrei R. Optimization of particle size and encapsulation efficiency of vancomycin nanoparticles by response surface methodology. Pharm Dev Technol 2014;19:987-98.
18. Pandit J, Sultana Y, Aqil M. Chitosan-coated PLGA nanoparticles of bevacizumab as novel drug delivery to target retina: Optimization, characterization, and in vitro toxicity evaluation. Artif Cells Nanomed Biotechnol 2017;45:1397-407.
19. Katamreddy JD, Yalavarthi PR, Rao DS, Teja SS, Battu S. In vitro characterization of statistically optimized quetiapine loaded self nanoemulsified systems with quality by design. Int J Pharm Invest 2018;8:14-3.
20. Abul Kalam M, Sultana Y, Ali A, Aqil M, Mishra AK, Aljuffali IA, et al. Part I: Development and optimization of solid-lipid nanoparticles using Box-Behnken statistical design for ocular delivery of gatifloxacin. J Biomed Mater Res A 2013;101:1813-27.
21. Rukmangathen R, Yallamalli IM, Yalavarthi PR. Formulation and biopharmaceutical evaluation of risperidone-loaded chitosan nanoparticles for intranasal delivery. Drug Dev Ind Pharm 2019;45:1342-50.
22. Ahmed IS, El-Hosary R, Shalaby S, Abd-Rabo MM, Elkhateeb DG, Nour S. PD-PK evaluation of freeze-dried atorvastatin calcium-loaded poly-?-caprolactone nanoparticles. Int J Pharm 2016;504:70-9.
23. Wang Y, Li P, Zheng P. Microencapsulation of nanoparticles with enhanced drug loading for pH-sensitive oral drug delivery for the treatment of colon cancer. J Appl Polym Sci 2013;129:714-20.
24. Woitiski CB, Neufeld RJ, Ribeiro AJ, Veiga F. Colloidal carrier integrating biomaterials for oral insulin delivery: Influence of component formulation on physicochemical and biological parameters. Acta Biomater 2009;5:2475-84.
25. Shah HA, Patel RP. Statistical modeling of zaltoprofen loaded biopolymeric nanoparticles: Characterization and anti-inflammatory activity of nanoparticles loaded gel. Int J Pharm Invest 2015;5:20-7.
26. Rahman Z, Zidan AS, Habib MJ, Khan MA. Understanding the quality of protein loaded PLGA nanoparticles variability by Plackett-Burman design. Int J Pharm 2010;389:186-94.
27. Chaves LL, Costa Lima SA, Vieira AC, Barreiros L, Segundo MA, Ferreira D, et al. pH-sensitive nanoparticles for improved oral delivery of dapsone: Risk assessment, design, optimization and characterization. Nanomedicine (Lond) 2017;12:1975-90.
28. Tayel AS, Mohamed A. Duodenum-triggered delivery of pravastatin sodium: II. Design, appraisal and pharmacokinetic assessments of enteric surface-decorated nanocubosomal dispersions. Drug Deliv 2016;23:3266-78.
29. Shailender J, Ravi PR, Reddy Sirukuri M, Dalvi A, Keerthi Priya O. Chitosan nanoparticles for the oral delivery of tenofovir disoproxil fumarate: Formulation optimization, characterization and ex vivo and in vivo evaluation for uptake mechanism in rats. Drug Dev Ind Pharm 2018;44:1109-19.
30. Rossato FA, Zecchin KG, La Guardia PG, Ortega RM, Alberici LC, Costa RA, et al. Fatty acid synthase inhibitors induce apoptosis in non-tumorigenic melan-a cells associated with inhibition of mitochondrial respiration. PLoS One 2014;9:e101060.
31. Sternby B, Hartmann D, Borgström B, Nilsson A. Degree of in vivo inhibition of human gastric and pancreatic lipases by Orlistat (Tetrahydrolipstatin, THL) in the stomach and small intestine. Clin Nutr 2002;21:395-402.
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How to Cite
BR, S. M., P. R. YELAVARTHI, and D. N. “PROCESS OF ORLISTAT-LOADED CHITOSAN NANOPARTICLES USING BOX–BEHNKEN DESIGN – AN EVALUATION STUDY”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 14, no. 5, May 2021, pp. 103-11, doi:10.22159/ajpcr.2021.v14i5.41441.
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