Pharmaceutical Research

, Volume 28, Issue 8, pp 2020–2033 | Cite as

Formulation, Biological and Pharmacokinetic Studies of Sucrose Ester-Stabilized Nanosuspensions of Oleanolic Acid

  • Wenji Li
  • Surajit Das
  • Ka-yun Ng
  • Paul W. S. Heng
Research Paper



The aim of this study was to develop sucrose ester (SE)-stabilized oleanolic acid (OA) nanosuspensions (NS) for enhanced delivery.


SEOA NS were prepared via O/W emulsion and organic solvent evaporation methods. The particles’ size and polydispersity index were measured by nanosizer. Their percent encapsulation efficiency, saturation solubility and in vitro dissolution rate were obtained via HPLC. The in vitro bioefficacy was analyzed by MTT measurements in A549 human non-small-cell lung cancer cell line. The cellular uptake of OA and in vivo pharmacokinetics profile were determined using LC-ESI-MS/MS.


Spherical SEOA NS particles (~100 nm in diameter) were produced and found to be physicochemically stable over a month at 4°C. In particular, SEOA 4121 NS (SEL: SEP at 4:1 w/w; SE: OA at 2:1 w/w) produced the greatest increase in saturation solubility (1.89 mg/mL vs. 3.43 μg/mL), dissolution rate, cytotoxicity and bioavailability. Preliminary studies indicated that cellular uptake of SEOA NS by A549 cells was temperature-, concentration- and time-dependent.


Preparing OA as SE-stabilized NS particles provides a novel method to enhance saturation solubility, in vitro dissolution rate, bioefficacy and in vivo bioavailability of free OA and/or other potentially useful hydrophobic drugs.


bioavailability dissolution rate hydrophobic drug oleanolic acid saturation solubility sucrose ester surfactant stabilized nanosuspensions 



This project is partially funded from GEA-NUS grant (N-148-000-008-001). The authors would like to thank Associate Professor HO Chi Lui, Paul, Department of Pharmacy, National University of Singapore, Singapore for his support in the DLS measurement. We thank Madam Loy Gek Luan and Mr. Chong Ping Lee, Department of Biological Sciences, National University of Singapore, Singapore for their support in EM experiments. We would also like to thank Compass Foods Pte. Ltd., Singapore, for the generous supply of SE.


  1. 1.
    Rasenack N, Hartenhauer H, Muller BW. Microcrystals for dissolution rate enhancement of poorly water-soluble drugs. Int J Pharm. 2003;254:137–45.PubMedCrossRefGoogle Scholar
  2. 2.
    Liang L, Tajmir-Riahi HA, Subirade M. Interaction of beta-lactoglobulin with resveratrol and its biological implications. Biomacromolecules. 2008;9:50–6.PubMedCrossRefGoogle Scholar
  3. 3.
    Betancor-Fernandez A, Perez-Galvez A, Sies H, Stahl W. Screening pharmaceutical preparations containing extracts of turmeric rhizome, artichoke leaf, devil's claw root and garlic or salmon oil for antioxidant capacity. J Pharm Pharmacol. 2003;55:981–6.PubMedCrossRefGoogle Scholar
  4. 4.
    Eibes G, Moreira MT, Feijoo G, Lema JM. Enzymatic degradation of low soluble compounds in monophasic water: solvent reactors. Kinetics and modeling of anthracene degradation by MnP. Biotechnol Bioeng. 2008;100:619–26.PubMedCrossRefGoogle Scholar
  5. 5.
    Kinder Jr FR, Versace RW, Bair KW, Bontempo JM, Cesarz D, Chen S, et al. Synthesis and antitumor activity of ester-modified analogues of bengamide B. J Med Chem. 2001;44:3692–9.PubMedCrossRefGoogle Scholar
  6. 6.
    Thiericke R. Drug discovery from nature: automated high-quality sample preparation. J Autom Methods Manag Chem. 2000;22:149–57.PubMedGoogle Scholar
  7. 7.
    Stefaniak AB, Chipera SJ, Day GA, Sabey P, Dickerson RM, Sbarra DC, et al. Physicochemical characteristics of aerosol particles generated during the milling of beryllium silicate ores: implications for risk assessment. J Toxicol Environ Health A. 2008;71:1468–81.PubMedCrossRefGoogle Scholar
  8. 8.
    Lanke SS, Gayakwad SG, Strom JG, and J. D’Souza M. Oral delivery of low molecular weight heparin microspheres prepared using biodegradable polymer matrix system. J Microencapsul. 2008;1–8 .Google Scholar
  9. 9.
    Chingunpitak J, Puttipipatkhachorn S, Chavalitshewinkoon-Petmitr P, Tozuka Y, Moribe K, Yamamoto K. Formation, physical stability and in vitro antimalarial activity of dihydroartemisinin nanosuspensions obtained by co-grinding method. Drug Dev Ind Pharm. 2008;34:314–22.PubMedCrossRefGoogle Scholar
  10. 10.
    Zhang D, Tan T, Gao L, Zhao W, Wang P. Preparation of azithromycin nanosuspensions by high pressure homogenization and its physicochemical characteristics studies. Drug Dev Ind Pharm. 2007;33:569–75.PubMedCrossRefGoogle Scholar
  11. 11.
    Chen Y, Liu J, Yang X, Zhao X, Xu H. Oleanolic acid nanosuspensions: preparation, in-vitro characterization and enhanced hepatoprotective effect. J Pharm Pharmacol. 2005;57:259–64.PubMedCrossRefGoogle Scholar
  12. 12.
    Pauland BK, Mitra RK. Percolation phenomenon in mixed reverse micelles: the effect of additives. J Colloid Interface Sci. 2006;295:230–42.CrossRefGoogle Scholar
  13. 13.
    Glatter O, Orthaber D, Stradner A, Scherf G, Fanun M, Garti N, et al. Sugar-Ester nonionic microemulsion: structural characterization. J Colloid Interface Sci. 2001;241:215–25.PubMedCrossRefGoogle Scholar
  14. 14.
    Honeywell-Nguyen PL, Frederik PM, Bomans PH, Junginger HE, Bouwstra JA. Transdermal delivery of pergolide from surfactant-based elastic and rigid vesicles: characterization and in vitro transport studies. Pharm Res. 2002;19:991–7.PubMedCrossRefGoogle Scholar
  15. 15.
    Grossiord JL, Thevenin MA, Poelman MC. Sucrose esters/cosurfactant microemulsion systems for transdermal delivery: assessment of bicontinuous structures. Int J Pharm. 1996;137:177–86.CrossRefGoogle Scholar
  16. 16.
    Kuntsche J, Koch MH, Steiniger F, Bunjes H. Influence of stabilizer systems on the properties and phase behavior of supercooled smectic nanoparticles. J Colloid Interface Sci. 2010;350:229–39.PubMedCrossRefGoogle Scholar
  17. 17.
    Klang V, Matsko N, Zimmermann AM, Vojnikovic E, Valenta C. Enhancement of stability and skin permeation by sucrose stearate and cyclodextrins in progesterone nanoemulsions. Int J Pharm. 2010;393:152–60.PubMedCrossRefGoogle Scholar
  18. 18.
    Takegami S, Kitamura K, Kawada H, Matsumoto Y, Kitade T, Ishida H, et al. Preparation and characterization of a new lipid nano-emulsion containing two cosurfactants, sodium palmitate for droplet size reduction and sucrose palmitate for stability enhancement. Chem Pharm Bull (Tokyo). 2008;56:1097–102.CrossRefGoogle Scholar
  19. 19.
    Ullrich S, Metz H, Mader K. Sucrose ester nanodispersions: microviscosity and viscoelastic properties. Eur J Pharm Biopharm. 2008;70:550–5.PubMedCrossRefGoogle Scholar
  20. 20.
    Arica Yegin B, Benoit JP, Lamprecht A. Paclitaxel-loaded lipid nanoparticles prepared by solvent injection or ultrasound emulsification. Drug Dev Ind Pharm. 2006;32:1089–94.PubMedCrossRefGoogle Scholar
  21. 21.
    Lippacher A, Muller RH, Mader K. Semisolid SLN dispersions for topical application: influence of formulation and production parameters on viscoelastic properties. Eur J Pharm Biopharm. 2002;53:155–60.PubMedCrossRefGoogle Scholar
  22. 22.
    Liu J, Liu Y, Parkinson A, Klaassen CD. Effect of oleanolic acid on hepatic toxicant-activating and detoxifying systems in mice. J Pharmacol Exp Ther. 1995;275:768–74.PubMedGoogle Scholar
  23. 23.
    Ovesna Z, Vachalkova A, Horvathova K, Tothova D. Pentacyclic triterpenoic acids: new chemoprotective compounds. Minireview. Neoplasma. 2004;51:327–33.PubMedGoogle Scholar
  24. 24.
    Konopleva M, Tsao T, Estrov Z, Lee RM, Wang RY, Jackson CE, et al. The synthetic triterpenoid 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid induces caspase-dependent and -independent apoptosis in acute myelogenous leukemia. Cancer Res. 2004;64:7927–35.PubMedCrossRefGoogle Scholar
  25. 25.
    Singh GB, Singh S, Bani S, Gupta BD, Banerjee SK. Anti-inflammatory activity of oleanolic acid in rats and mice. J Pharm Pharmacol. 1992;44:456–8.PubMedCrossRefGoogle Scholar
  26. 26.
    Gao D, Li Q, Li Y, Liu Z, Liu Z, Fan Y, et al. Antidiabetic potential of oleanolic acid from Ligustrum lucidum Ait. Can J Physiol Pharmacol. 2007;85:1076–83.PubMedCrossRefGoogle Scholar
  27. 27.
    Ji HY, Shin BS, Jeong DW, Park EJ, Park ES, Yoo SD, et al. Interspecies scaling of oleanolic acid in mice, rats, rabbits and dogs and prediction of human pharmacokinetics. Arch Pharm Res. 2009;32:251–7.PubMedCrossRefGoogle Scholar
  28. 28.
    Jeong DW, Kim YH, Kim HH, Ji HY, Yoo SD, Choi WR, et al. Dose-linear pharmacokinetics of oleanolic acid after intravenous and oral administration in rats. Biopharm Drug Dispos. 2007;28:51–7.PubMedCrossRefGoogle Scholar
  29. 29.
    Song M, Hang TJ, Wang Y, Jiang L, Wu XL, Zhang Z, et al. Determination of oleanolic acid in human plasma and study of its pharmacokinetics in Chinese healthy male volunteers by HPLC tandem mass spectrometry. J Pharm Biomed Anal. 2006;40:190–6.PubMedCrossRefGoogle Scholar
  30. 30.
    Gao Z, Maloney DJ, Dedkova LM, Hecht SM. Inhibitors of DNA polymerase beta: activity and mechanism. Bioorg Med Chem. 2008;16:4331–40.PubMedCrossRefGoogle Scholar
  31. 31.
    Das S, Lin HS, Ho PC, Ng KY. The impact of aqueous solubility and dose on the pharmacokinetic profiles of resveratrol. Pharm Res. 2008;25:2593–600.PubMedCrossRefGoogle Scholar
  32. 32.
    Patravale VB, Date AA, Kulkarni RM. Nanosuspensions: a promising drug delivery strategy. J Pharm Pharmacol. 2004;56:827–40.PubMedCrossRefGoogle Scholar
  33. 33.
  34. 34.
    SUCROSE MONOLAURATE - Compound Summary (CID 4182596)
  35. 35.
    Noyesand A, Whitney W. The rate of solution of solid substances in their own solutions. J Am Chem Soc. 1897;19:930–4.CrossRefGoogle Scholar
  36. 36.
    Carey HV, Andrews MT, Martin SL. Mammalian hibernation: cellular and molecular responses to depressed metabolism and low temperature. Physiol Rev. 2003;83:1153–81.PubMedGoogle Scholar
  37. 37.
    Chan OH, Stewart BH. Physicochemical and drug-delivery considerations for oral drug bioavailability. Drug Discov Today. 1996;1:12.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Wenji Li
    • 1
  • Surajit Das
    • 1
  • Ka-yun Ng
    • 1
  • Paul W. S. Heng
    • 1
  1. 1.Department of PharmacyNational University of SingaporeSingaporeSingapore

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