Design and evaluation of a novel nanoparticulate-based formulation encapsulating a HIP complex of lysozyme

Formulation development of protein therapeutics using polymeric nanoparticles has found very little success in recent years. Major formulation challenges include rapid denaturation, susceptibility to lose bioactivity in presence of organic solvents and poor encapsulation in polymeric matrix. In the present study, we have prepared hydrophobic ion pairing (HIP) complex of lysozyme, a model protein, using dextran sulfate (DS) as a complexing polymer. We have optimized the process of formation and dissociation of HIP complex between lysozyme and DS. The effect of HIP complexation on enzymatic activity of lysozyme was also studied. Nanoparticles were prepared and characterized using spontaneous emulsion solvent diffusion method. Furthermore, we have also investigated release of lysozyme from nanoparticles along with its enzymatic activity. Results of this study indicate that nanoparticles can sustain the release of lysozyme without compromising its enzymatic activity. HIP complexation using a polymer may also be employed to formulate sustained release dosage forms of other macromolecules with enhanced encapsulation efficiency.

[1]  R. Sridhar,et al.  Inactivation of lysozyme by sonication under conditions relevant to microencapsulation. , 2000, International journal of pharmaceutics.

[2]  Daniel E. Otzen,et al.  Protein drug stability: a formulation challenge , 2005, Nature Reviews Drug Discovery.

[3]  B. Sarmento,et al.  Development and characterization of new insulin containing polysaccharide nanoparticles. , 2006, Colloids and surfaces. B, Biointerfaces.

[4]  Steven P Schwendeman,et al.  Recent advances in the stabilization of proteins encapsulated in injectable PLGA delivery systems. , 2002, Critical reviews in therapeutic drug carrier systems.

[5]  W. Tiyaboonchai,et al.  Formulation and characterization of amphotericin B-chitosan-dextran sulfate nanoparticles. , 2007, International journal of pharmaceutics.

[6]  E. Topp,et al.  Chemical degradation of peptides and proteins in PLGA: a review of reactions and mechanisms. , 2008, Journal of pharmaceutical sciences.

[7]  D. Sacco,et al.  Interaction of a macromolecular polyanion, dextran sulfate, with human hemoglobin , 1986, FEBS letters.

[8]  A. Schaper,et al.  Charged nanoparticles as protein delivery systems: a feasibility study using lysozyme as model protein. , 2008, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[9]  H. Benson,et al.  Development of a chitosan-based nanoparticle formulation for delivery of a hydrophilic hexapeptide, dalargin. , 2008, Biopolymers.

[10]  K. Dill,et al.  Charge effects on folded and unfolded proteins. , 1990, Biochemistry.

[11]  S. Frokjaer,et al.  Preparing and evaluating delivery systems for proteins. , 2006, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[12]  S. Nie,et al.  Therapeutic Nanoparticles for Drug Delivery in Cancer Types of Nanoparticles Used as Drug Delivery Systems , 2022 .

[13]  E. Allémann,et al.  Nanoprecipitation versus emulsion-based techniques for the encapsulation of proteins into biodegradable nanoparticles and process-related stability issues , 2005, AAPS PharmSciTech.

[14]  A. Mitra,et al.  Encapsulation of Protein-Polysaccharide HIP Complex in Polymeric Nanoparticles , 2011, Journal of drug delivery.

[15]  Weien Yuan,et al.  Formulating protein therapeutics into particulate forms , 2009, Expert opinion on drug delivery.

[16]  H. Itagaki,et al.  Hemoglobin denaturation caused by surfactants. , 1995, Biological & pharmaceutical bulletin.

[17]  J. R. Amrutkar,et al.  Chitosan–Chondroitin Sulfate Based Matrix Tablets for Colon Specific Delivery of Indomethacin , 2009, AAPS PharmSciTech.

[18]  R. Hurwitz,et al.  Painful purpura: an adverse effect to a thrombolysin. , 1990, Archives of dermatology.

[19]  Brian M. Murphy,et al.  Stability of Protein Pharmaceuticals: An Update , 2010, Pharmaceutical Research.

[20]  Wim E. Hennink,et al.  Protein Instability in Poly(Lactic-co-Glycolic Acid) Microparticles , 2000, Pharmaceutical Research.

[21]  B. Sarmento,et al.  Lipid-based colloidal carriers for peptide and protein delivery – liposomes versus lipid nanoparticles , 2007, International journal of nanomedicine.

[22]  M. Manning,et al.  Hydrophobic Ion Pairing: Altering the Solubility Properties of Biomolecules , 1998, Pharmaceutical Research.

[23]  Dongmei Cun,et al.  Design of high payload PLGA nanoparticles containing melittin/sodium dodecyl sulfate complex by the hydrophobic ion-pairing technique , 2009, Drug development and industrial pharmacy.

[24]  S. Pispas,et al.  Sustained and extended release with structural and activity recovery of lysozyme from complexes with sodium (sulfamate carboxylate) isoprene/ethylene oxide block copolymer. , 2010, Macromolecular bioscience.

[25]  L. Dong,et al.  Characterization of physiochemical and biological properties of an insulin/lauryl sulfate complex formed by hydrophobic ion pairing. , 2007, International journal of pharmaceutics.

[26]  B. Sunderland,et al.  An ion pairing approach to increase the loading of hydrophilic and lipophilic drugs into PEGylated PLGA nanoparticles. , 2009, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[27]  D. Mcclements Non-covalent interactions between proteins and polysaccharides. , 2006, Biotechnology advances.

[28]  K. Kisich,et al.  The potential advantages of nanoparticle drug delivery systems in chemotherapy of tuberculosis. , 2005, American Journal of Respiratory and Critical Care Medicine.

[29]  T. Park,et al.  Protein-fatty acid complex for enhanced loading and stability within biodegradable nanoparticles. , 2001, Journal of pharmaceutical sciences.

[30]  Hong Yuan,et al.  Strategic approaches for improving entrapment of hydrophilic peptide drugs by lipid nanoparticles. , 2009, Colloids and surfaces. B, Biointerfaces.

[31]  E. Leo,et al.  Fabrication via a nonaqueous nanoprecipitation method, characterization and in vitro biological behavior of N(6)-cyclopentyladenosine-loaded nanoparticles. , 2009, Journal of pharmaceutical sciences.

[32]  C. F. van der Walle,et al.  Current approaches to stabilising and analysing proteins during microencapsulation in PLGA. , 2009, Expert opinion on drug delivery.