Purpose:

Sulfasalazine has recently been identified to inhibit the cell growth in neuroblastoma cell lines via sepiapterin reductase (SPR) inhibition and the potent inhibitory effect on the x−c cystine transporter blocking the uptake of cysteine by the cells. It was originally synthesized as a prodrug for the treatment of the Rheumatoid Arthritis and has also been used in treatment of the Crohn’s disease. The drug shows about 10-30% bioavailability on oral administration and is metabolized by bacterial azoreductases in the two metabolites namely, sulfapyridine and mesalazine. Hence for the delivery of the drug to the tumor, bypassing the metabolism as well as increased bioavailability is necessary. Also, Sulfasalazine has been shown to have low cell penetrability preventing effective use of the sulfasalazine in free drug form. The objective of the study was to formulate the sulfasalazine loaded albumin-chitosan hybrid nanocarriers with particle size less than 100 nm and evaluate them against neuroblastoma cells which can be administered systemically to effectively overcome the challenges posed by the drug.

 

Methods: In this study, we developed a nano-precipitation method to produce the self-assembling hydrophobic drug Sulfasalazine-loaded Bovine Serum Albumin (BSA) nanocarriers. The experiments were designed using the Taguchi orthogonal array, and an optimal formulation was selected to obtain the drug loading of > 5 %, particle size of   The nanocarriers were prepared using 2% BSA solution and 74% acetone in the ratio 1:5.4. The formulation was kept for evaporation of solvent for 17 hrs while stirring at 600rpm. The 5mg/ml chitosan solution was added in a ratio of 1:10 to the total volume of the BSA nanocarriers. Next, the nanocarriers were PEGylated using the activated mPEG-SPA to impart salt and serum stability. The nanocarriers were characterized using dynamic light scattering particle size analysis and zeta potential analysis. Sulfasalazine was quantified at 360 nm using a plate reader. The drug release study in a simulated body fluid of pH 7.4 was performed to evaluate the release profile of the drug from the nanocarriers. In addition, the nanocarriers were subjected to stability testing under different storage conditions.


Results: The Sulfasalazine-loaded PEGylated nanocarriers had an average particle size of 80.556 ± 3.119 nm with an average PDI of 0.120 ± 0.037 and zeta potential of the formulation was found to be 5.593 ± 0.583mV. The entrapment efficiency of the formulation was found to be 98.65%.  The achieved drug loading for the final formulation was about 6.78% (w/w). In the release study, the developed nanocarriers showed the controlled release of the drug up to 48 hrs in a simulated body fluid of pH 7.4. The nanocarriers were found to be stable in the presence of PBS 7.4 and 55% FBS solution for more than 72 hrs. Currently, the developed nanocarriers are being tested for dose-dependent anticancer effects against neuroblastoma cell lines.


Conclusion:

Stable Sulfasalazine-loaded nanocarriers of < 100 nm with an average PDI of 0.120 ± 0.037 were developed. Sulfasalazine-loaded nanocarriers showed controlled release of the loaded drug. This investigation infers that albumin-chitosan polymer-based nanocarriers are useful for the delivery of anticancer drugs. The results for the prepared formulation are encouraging and suggest further analysis of these nanocarriers by in vitro studies in cancer cells and in vivo studies using neuroblastoma tumor-bearing mice.

Presenting Author(s)

• SM

  Sushrut Marathe

  – University of Mississippi, Oxford

Main Author(s)

• SM

  Sushrut Marathe

  – University of Mississippi, Oxford

Co-Author(s)

• RJ

  Rohit Joshi

  – post doc`, University of Mississippi, oxford, Mississippi
• AB

  Andre Bachmann

  – Michigan State University

Primary/Principal Investigator(s)

• MC

  Mahavir Bhupal Chougule

  – Associate Professor, University of Mississippi, Mississippi

Co-Author(s)

• MC

  Mahavir Bhupal Chougule

  – Associate Professor, University of Mississippi, Mississippi
• SM

  Sushrut Marathe

  – University of Mississippi, Oxford