Application of Box-Behnken design for optimization of formulation parameters for nanostructured lipid carriers of candesartan cilexetil

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Hetal P. Thakkar
Jagruti L. Desai
Mayur P. Parmar


This study deals with development and optimization of nanostructured lipid carriers (NLCs) of candesartan cilexetil (CC)
for improving its oral bioavailability. From solubility and lipid‑water partition studies of CC in various lipids, glyceryl
monostearate (GMS) and glyceryl monocaprylate were selected as solid lipid and liquid lipid, respectively. NLCs were
formulated by hot melt‑emulsification‑ultrasonication method. A three‑factor, three‑level Box–Behnken design was used to
optimize the independent variables, lipid: drug ratio (X1), solid lipid: liquid lipid ratio (X2) and surfactant concentration (X3).
Different batches were prepared and evaluated for responses, particle size (Y1), zeta potential (Y2) and % entrapment
efficiency (Y3). Response surface plots and perturbation plots were constructed to study the effect of factors on responses.
The optimized formulation containing X1 - 22.47:1, X2 - 7.23:1 and X3 - 1.97% was prepared and evaluated. Observed
values for Y1, Y2, and Y3 were found to be closer to the predicted values thus validating the optimization method. Differential
scanning calorimetry thermograms of pure drug, GMS and lyophilized drug loaded NLCs indicated complete miscibility
of drug into lipids. The release of CC from the NLCs conducted in artificial gastric fluid (pH 1.2) was much higher than in
phosphate buffer solution (pH 6.8). The formulated NLCs were found to be more stable at refrigerated condition (5°C ± 3°C)
as compared with room temperature (25°C ± 2°C/60% RH% ± 5% RH). The use of design approach helped to identify
critical formulation parameters in CC loaded NLCs preparation.
Key words: Candesartan, hypertension, nanostructured lipid carriers, oral bioavailability partition study


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Thakkar, H. P., Desai, J. L., & Parmar, M. P. (2014). Application of Box-Behnken design for optimization of formulation parameters for nanostructured lipid carriers of candesartan cilexetil. Asian Journal of Pharmaceutics (AJP), 8(2).


Kearney PM, Whelton M, Reynolds K, Muntner P, Whelton PK, He J.

Global burden of hypertension: Analysis of worldwide data. Lancet


High blood pressure and cholesterol: out of control in US.

Atlanta. Available from:

CardiovascularDisease. [Last cited on 2013 Apr 25].

Tadevosyan A, Maclaughlin EJ, Karamyan VT. Angiotensin II type 1

receptor antagonists in the treatment of hypertension in elderly

patients: Focus on patient outcomes. Patient Relat Outcome Meas


David PW. Antihypertensive drugs. In: Charles R, Craig RE, editors.

Modern Pharmacology with Clinical Applications. Beltimore: Lippincott

Williams and Wilkins; 1997. p. 225‑38.

Ferreiros N, Dresen S, Alonso RM, Weinmann W. Hydrolysis and

transesterification reactions of candesartan cilexetil observed during

the solid phase extraction procedure. J Chromatogr B Analyt Technol

Biomed Life Sci 2007;855:134‑8.

Nishida Y, Takahashi Y, Nakayama T, Soma M, Asai S. Comparative effect

of olmesartan and candesartan on lipid metabolism and renal function

in patients with hypertension: A retrospective observational study.

Cardiovasc Diabetol 2011;10:74.

Nekkanti V, Karatgi P, Prabhu R, Pillai R. Solid self‑microemulsifying

formulation for candesartan cilexetil. AAPS PharmSciTech 2010;11:9‑17.

Al Omari AA, Al Omari MM, Badwan AA, Al‑Sou’od KA. Effect of

cyclodextrins on the solubility and stability of candesartan cilexetil in

solution and solid state. J Pharm Biomed Anal 2011;54:503‑9.

Sayyad FJ, Tulsankar SL, Kolap UB. Design and development of liquisolid

compact of candesartan cilexetil to enhance dissolution. J Pharm Res


Zhang Z, Gao F, Bu H, Xiao J, Li Y. Solid lipid nanoparticles loading

candesartan cilexetil enhance oral bioavailability: In vitro characteristics

and absorption mechanism in rats. Nanomedicine 2012;8:740‑7.

Gao F, Zhang Z, Bu H, Huang Y, Gao Z, Shen J, et al. Nanoemulsion

improves the oral absorption of candesartan cilexetil in rats:

Performance and mechanism. J Control Release 2011;149:168‑74.

Burcham DL, Maurin MB, Hausner EA, Huang SM. Improved oral

bioavailability of the hypocholesterolemic DMP 565 in dogs following

oral dosing in oil and glycol solutions. Biopharm Drug Dispos


Shafiq S, Shakeel F, Talegaonkar S, Ahmad FJ, Khar RK, Ali M.

Development and bioavailability assessment of ramipril nanoemulsion

formulation. Eur J Pharm Biopharm 2007;66:227‑43.

Venishetty VK, Chede R, Komuravelli R, Adepu L, Sistla R, Diwan PV.

Design and evaluation of polymer coated carvedilol loaded solid lipid

nanoparticles to improve the oral bioavailability: A novel strategy

to avoid intraduodenal administration. Colloids Surf B Biointerfaces


Severino P, Andreani T, Macedo AS, Fangueiro JF, Santana MH, Silva AM,

et al. Current state‑of‑art and new trends on lipid nanoparticles (SLN

and NLC) for oral drug delivery. J Drug Deliv 2012;2012:750891.

Müller RH, Mäder K, Gohla S. Solid lipid nanoparticles (SLN) for

controlled drug delivery‑A review of the state of the art. Eur J Pharm

Biopharm 2000;50:161‑77.

Muchow M, Maincent P, Muller RH. Lipid nanoparticles with a solid

matrix (SLN, NLC, LDC) for oral drug delivery. Drug Dev Ind Pharm


Kasongo KW, Shegokar R, Müller RH, Walker RB. Formulation

development and in vitro evaluation of didanosine‑loaded nanostructured

lipid carriers for the potential treatment of AIDS dementia complex.

Drug Dev Ind Pharm 2011;37:396‑407.

Gohel MC, Amin AF. Formulation optimization of controlled release

diclofenac sodium microspheres using factorial design. J Control

Release 1998;51:115‑22.

Nazzal S, Khan MA. Response surface methodology for the optimization

of ubiquinone self‑nanoemulsified drug delivery system. AAPS Pharm

Sci Tech 2002;3:E3.

Gareth A. Lewis DM, Phan‑Tan‑Luu R. Pharmaceutical Experimental

Design. New York, Basel: Marcel Deckker Inc.; 2005.

Joshi M, Patravale V. Formulation and evaluation of nanostructured

lipid carrier (NLC)‑based gel of valdecoxib. Drug Dev Ind Pharm


Doktorovová S, Araújo J, Garcia ML, Rakovský E, Souto EB. Formulating

fluticasone propionate in novel PEG‑containing nanostructured lipid

carriers (PEG‑NLC). Colloids Surf B Biointerfaces 2010;75:538‑42.

Aji Alex MR, Chacko AJ, Jose S, Souto EB. Lopinavir loaded solid lipid

nanoparticles (SLN) for intestinal lymphatic targeting. Eur J Pharm Sci


Box GE, Behnken DW. Some new three‑level designs for the study of

quantitative variables. Technometrics 1960;2:455‑75.

Dong CH, Xie XQ, Wang XL, Zhan Y, Yao YJ. Application of

Box‑Behnken design in optimisation for polysaccharides extraction

from cultured mycelium of Cordyceps sinensis. Food Bioprod Process


Gulbake A, Jain A, Khare P, Jain SK. Solid lipid nanoparticles bearing

oxybenzone: In‑vitro and in‑vivo evaluation. J Microencapsul


Unruh T, Bunjes H, Westsen K. Observation of size‑dependent melting

in lipid nanoparticles. J Phys Chem 1999;103:10373‑7.

ICH Q 1 A (R2). Stability Testing of New Drug Substances and

Products (CPMP/ICH/2736/99); 2003.

Gonzalez‑Mira E, Egea MA, Garcia ML, Souto EB. Design and ocular

tolerance of flurbiprofen loaded ultrasound‑engineered NLC. Colloids

Surf B Biointerfaces 2010;81:412‑21.

Uner M, Yener G. Importance of solid lipid nanoparticles (SLN) in various

administration routes and future perspectives. Int J Nanomedicine


Woitiski CB, Veiga F, Ribeiro A, Neufeld R. Design for optimization of

nanoparticles integrating biomaterials for orally dosed insulin. Eur J

Pharm Biopharm 2009;73:25‑33.

Gannu R, Yamsani VV, Yamsani SK, Palem CR, Yamsani MR. Optimization

of hydrogels for transdermal delivery of lisinopril by Box‑Behnken

statistical design. AAPS Pharm Sci Tech 2009;10:505‑14.

Jia LJ, Zhang DR, Li ZY, Feng FF, Wang YC, Dai WT, et al. Preparation

and characterization of silybin‑loaded nanostructured lipid carriers.

Drug Deliv 2010;17:11‑8.

Finney J. The structural basis of the hydrohobic interaction.

In: Bellissent‑Funel MC, editor. Hydration Processes in Biology:

Theoretical and Experimental Approaches. France: IOS Press; 1998.

p. 115‑26.

Han F, Sanming L, Yin R, Liu H, Xu L. Effect of surfactants on the

formation and characterization of a new type of colloidal drug delivery

system: Nanostructured lipid carriers. Colloids Surf A Physicochem Eng

Asp 2008;315:210‑6.

Unruh T, Bunjes H, Westesen K, Koch MH, Investigations on the

melting behaviour of triglyceride nanoparticles. Colloid Polym Sci


Nekkanti V, Pillai R, Venkateshwarlu V, Harisudhan T. Development and

characterization of solid oral dosage form incorporating candesartan

nanoparticles. Pharm Dev Technol 2009;14:290‑8.

Chakraborty S, Shukla D, Mishra B, Singh S. Lipid – An emerging

platform for oral delivery of drugs with poor bioavailability. Eur J Pharm

Biopharm 2009;73:1‑15.

Beloqui A, Solinís MÃ, Delgado A, Evora C, Isla A, Rodríguez‑Gascón A.

Fate of nanostructured lipid carriers (NLCs) following the oral route:

Design, pharmacokinetics and biodistribution. J Microencapsul


Joshi M, Patravale V. Nanostructured lipid carrier (NLC) based gel of

celecoxib. Int J Pharm 2008;346:124‑32.