Abstract
Purpose
An optimized methodology for the development of a new generation of lipid nanoparticles, the multiple lipid nanoparticles (MLN) is described. MLN have characteristics between nanostructured lipid carriers (NLC) and multiple emulsions (W/O/W), but without the outer aqueous phase.
Methods
The production is based on a hot homogenization method combined with high shear and ultrasonication. The antiretroviral agent lamivudine (3TC), was loaded in the MLN. For comparison purposes, NLC-3TC formulation was also developed and physico-chemically characterized by the same parameters as MLN-3TC. The development and optimization of MLN and NLC formulations were supported by a Quality by Design (QbD) approach.
Results
The MLN-3TC formulation exhibited a size of about 450 nm, polydispersity <0.3 and negative zeta potential > −20 mV. Furthermore, the morphology assessed by TEM showed a structure with multiples aqueous vacuoles. MLN-3TC was physically stable for at least 45 days, had low cytotoxicity and drug release studies showed a sustained and controlled release of 3TC under gastric and plasma-simulated conditions (at pH 7.4 for about 45 h).
Conclusions
The optimized formulations present suitable profiles for oral administration. Overall, the results reveal that MLN present higher loading capacity and storage stability than NLC.
Similar content being viewed by others
Abbreviations
- 3TC:
-
Lamivudine
- ANOVA:
-
Analysis of variance
- AP:
-
Aqueous phase
- BCS:
-
Biopharmaceutical classification system
- DLS:
-
Dynamic light scattering
- DMSO:
-
Dimethyl sulfoxide
- DOE:
-
Design of experiments
- EE:
-
Entrapment efficiency
- EPC:
-
Egg phosphatidylcholine
- FBS:
-
Fetal bovine serum
- FMEA:
-
Failure modes and effects analysis
- HIV:
-
Human immunodeficiency virus
- HLB:
-
Hydrophilic-lipophilic balance
- IC50 :
-
Half maximal inhibitory concentration
- LC:
-
Loading capacity
- LL:
-
Liquid lipid
- LNP:
-
Lipid nanoparticles
- MLN:
-
Multiple lipid nanoparticles
- MLP:
-
Multiple lipid particles
- MTT:
-
Methylthiazoletetrazolium
- NLC:
-
Nanostructured lipid carriers
- NPs:
-
Nanoparticles
- NRTI:
-
Reverse transcriptase inhibitors
- PDI:
-
Polydispersity index
- QbD:
-
Quality by design
- RS:
-
Response surface
- RT:
-
Room temperature
- SL:
-
Solid lipid
- SLN:
-
Solid lipid nanoparticles
- TEM:
-
Transmission electron microscopy
- W/O/W:
-
Water/oil/water
- Zeta:
-
Zeta potential
References
Battaglia L, Gallarate M. Lipid nanoparticles: state of the art, new preparation methods and challenges in drug delivery. Expert Opin Drug Deliv. 2012;9(5):497–508.
Das S, Chaudhury A, PharmSciTech A. Recent advances in lipid nanoparticle formulations with solid matrix for oral drug delivery. AAPS PharmSciTech. 2011;12(1):62–76.
Pardeike J, Hommoss A, Müller RH. Lipid nanoparticles (SLN, NLC) in cosmetic and pharmaceutical dermal products. Int J Pharm. 2009;366(1–2):170–84.
Severino P, Andreani T, Macedo AS, Fangueiro JF, Santana MHA, Silva AM, Souto EB. Current state-of-art and new trends on lipid nanoparticles (SLN and NLC) for oral drug delivery. J Drug Deliv. 2012;2012:10–0.
Gasco MR. Method for producing solid lipid microspheres having a narrow size distribution. In.: Google Patents. 1993.
Lucks S, Müller R. Medication vehicles made of solid lipid particles (solid lipid nanospheres - sln). In.: Google Patents. 1996.
Ali Khan A, Mudassir J, Mohtar N, Darwis Y. Advanced drug delivery to the lymphatic system: lipid-based nanoformulations. Int J Nanomedicine. 2013;8:2733–44.
Muchow M, Maincent P, Müller RH. Lipid nanoparticles with a solid matrix (SLN®, NLC®, LDC®) for oral drug delivery. Drug Dev Ind Pharm. 2008;34(12):1394–405.
Zhao G, Hu C, Sun R, Ni S, Li Q, Xia Q. Development of novel composite antioxidant multiple lipid particles from combination of W/O/W multiple emulsions and solid lipid nanoparticles. Eur J Lipid Sci Technol. 2015;117(7):1056–65.
Chaturvedi SP, Kumar V. Production techniques of lipid nanoparticles: a review. Res J Pharm, Biol Chem Sci. 2012;3(3):16.
ICH. Quality risk management Q9. In.: International conference on harmonisation of technical requirements for registration of pharmaceuticals for human use; 2005.
ICH. Pharmaceutical development - Q8(R2). In.: International conference on harmonisation of technical requirements for registration of pharmaceuticals for human use; 2009.
Benício de Barros Neto Roy Edward Bruns ISS, Bookman. Como fazer experimentos: Bookman; 2010.
ICH. Pharmaceutical quality system - Q10. In. http://www.fda.gov/downloads/Drugs/.../Guidances/ucm073517.pdf: International conference on harmonisation of technical requirements for registration of pharmaceuticals for human use; 2009.
Maria Isabel Rodrigues AFL, Cárita. Planejamento de Experimentos e Otimização de Precessos: Cárita; 2014.
Bedse G, Kumar V, Singh S. Study of forced decomposition behavior of lamivudine using LC, LC–MS/TOF and MSn. J Pharm Biomed Anal. 2009;49(1):55–63.
Strauch S, Jantratid E, Dressman JB, Junginger HE, Kopp S, Midha KK, Shah VP, Stavchansky S, Barends DM. Biowaiver monographs for immediate release solid oral dosage forms: lamivudine. J Pharm Sci. 2011;100(6):2054–63.
Lindenberg M, Kopp S, Dressman JB. Classification of orally administered drugs on the World Health Organization model list of essential medicines according to the biopharmaceutics classification system. Eur J Pharm Biopharm. 2004;58(2):265–78.
Vyas TK, Shah L, Amiji MM. Nanoparticulate drug carriers for delivery of HIV/AIDS therapy to viral reservoir sites. Expert Opin Drug Deliv. 2006;3(5):613–28.
Ferreira M, Chaves LL, Lima SAC, Reis S. Optimization of nanostructured lipid carriers loaded with methotrexate: a tool for inflammatory and cancer therapy. Int J Pharm. 2015;492(1–2):65–72.
Griffin WC. Classification of surface-active agents by “HLB”. J Soc Cosmet Chem. 1949;1:311–26.
Griffin WC. Calculation of HLB values of non-ionic surfactants. J Soc Cosmet Chem. 1954;5:249–56.
Martins S, Tho I, Reimold I, Fricker G, Souto E, Ferreira D, Brandl M. Brain delivery of camptothecin by means of solid lipid nanoparticles: formulation design, in vitro and in vivo studies. Int J Pharm. 2012;439(1–2):49–62.
Schöler N, Hahn H, Müller RH, Liesenfeld O. Effect of lipid matrix and size of solid lipid nanoparticles (SLN) on the viability and cytokine production of macrophages. Int J Pharm. 2002;231(2):167–76.
Block ID. Particle sizing by 3D cross-correlation DLS in highly scattering samples. In. http://www.lsinstruments.ch/__/frontend/handler/document.php?id=265&type=42: LS Instruments; 2010.
Lippacher A, Müller RH, Mäder K. Preparation of semisolid drug carriers for topical application based on solid lipid nanoparticles. Int J Pharm. 2001;214(1–2):9–12.
Mitri K, Shegokar R, Gohla S, Anselmi C, Müller RH. Lipid nanocarriers for dermal delivery of lutein: preparation, characterization, stability and performance. Int J Pharm. 2011;414(1–2):267–75.
S-s F, Huang G. Effects of emulsifiers on the controlled release of paclitaxel (Taxol®) from nanospheres of biodegradable polymers. J Control Release. 2001;71(1):53–69.
Wissing SA, Muller RH. Solid lipid nanoparticles as carrier for sunscreens: in vitro release and in vivo skin penetration. J Control Release. 2002;81(3):225–33.
Mishra PR, Shaal LA, Müller RH, Keck CM. Production and characterization of Hesperetin nanosuspensions for dermal delivery. Int J Pharm 2009;371(1–2):182–189.
Souto EB, Wissing SA, Barbosa CM, Müller RH. Evaluation of the physical stability of SLN and NLC before and after incorporation into hydrogel formulations. Eur J Pharm Biopharm. 2004;58(1):83–90.
Ekambaram P, Satahali AAH, Priyanka K. Solid lipid nanoparticles: review. Scientific Reviews Chemical Communications. 2012;2(1):80–102.
Frias I, Neves AR, Pinheiro M, Reis S. Design, development, and characterization of lipid nanocarriers-based epigallocatechin gallate delivery system for preventive and therapeutic supplementation. Drug Des Devel Ther. 2016;10:3519–28.
Sarmento B, Martins S, Ferreira D, Souto EB. Oral insulin delivery by means of solid lipid nanoparticles. Int J Nanomedicine. 2007;2(4):743–9.
Shah M, Agrawal YK, Garala K, Ramkishan A. Solid lipid nanoparticles of a water soluble drug, ciprofloxacin hydrochloride. Indian J Pharm Sci. 2012;74(5):434–42.
Singh S, Dobhal AK, Jain A, Pandit JK, Chakraborty S. Formulation and evaluation of solid lipid nanoparticles of a water soluble drug: zidovudine. Chem Pharm Bull. 2010;58(5):650–5.
Muller RH, Becker R, Kruss B, Peters K. Pharmaceutical nanosuspensions for medicament administration as systems with increased saturation solubility and rate of solution. In.: Google Patents; 1999.
Zur Muhlen A, Mehnert W. Drug release and release mechanism of prednisolone loaded solid lipid nanoparticles. Pharmazie. 1998;53(8)
Mehnert W, Mäder K. Solid lipid nanoparticles: production, characterization and applications. Adv Drug Deliv Rev. 2001;47(2–3):165–96.
Dubes A, Parrot-Lopez H, Abdelwahed W, Degobert G, Fessi H, Shahgaldian P, Coleman AW. Scanning electron microscopy and atomic force microscopy imaging of solid lipid nanoparticles derived from amphiphilic cyclodextrins. Eur J Pharm Biopharm. 2003;55(3):279–82.
Li Q, Du Y-Z, Yuan H, Zhang X-G, Miao J, Cui F-D, Hu F-Q. Synthesis of lamivudine stearate and antiviral activity of stearic acid-g-chitosan oligosaccharide polymeric micelles delivery system. Eur J Pharm Sci. 2010;41(3–4):498–507.
Acknowledgments and Disclosures
Cavalcanti and Soares-Sobrinho thank the Facepe (1316–4.03/12) and CNPq (482,954/2013–2) for financial support. Lima thanks Operação NORTE-01-0145-FEDER-000011. Nunes thanks FCT for her Investigator grant (IF/00293/2015). The authors are also grateful to Dr. Rui Fernandes (Histology and Electron Microscopy Service – Instituto de Biologia Molecular e Celular [HEMS-I3S], University of Porto). This work received financial support from the European Union (FEDER funds) and National Funds (FCT/MEC, Fundação para a Ciência e Tecnologia and Ministério da Educação e Ciência) under the Partnership Agreement PT2020 UID/MULTI/04378/2013.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
ESM 1
(PDF 1.25 MB)
Rights and permissions
About this article
Cite this article
Cavalcanti, S.M.T., Nunes, C., Lima, S.A.C. et al. Multiple Lipid Nanoparticles (MLN), a New Generation of Lipid Nanoparticles for Drug Delivery Systems: Lamivudine-MLN Experimental Design. Pharm Res 34, 1204–1216 (2017). https://doi.org/10.1007/s11095-017-2136-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11095-017-2136-0