In silico pipeline for nanoparticle optimized formulations to immunobiological delivery

Introduction: Pharmaceutical nanotechnology can improve biopharmaceutical therapies by overcoming common stability, safety, and pharmacokinetics challenges.Objectives: This study utilized a bioactive peptide with promising therapeutic applications as an initial model, modifying it to increase hydrophobicity and improve nanoparticle encapsulation efficiency.In a subsequent phase of the research, we plan to assess alterations in binding affinity through molecular docking and molecular dynamics simulations.Methodology: Hydrophobicity was estimated using the grand average of hydropathy (GRA VY) index, and most hydrophobic sequences were selected for the in-silico safety evaluation.Toxicity, allergenicity, aggregation, and immunogenicity analyses were conducted using the ToxinPred 3.0, AlgPred 2.0, Pasta 2.0, BEPIPRED 3.0, and NetMHCIIpan -4.1 predictors.Results: A total of 23 sequence modifications were proposed.GRA VY index estimations revealed seven sequences with greater hydrophobicity than the original sequence (>0.25).The most hydrophobic (0.31) and hydrophilic (-0.26) sequences were selected for encapsulation efficiency tests.Safety analysis of the original sequence indicated non-toxicity (hybrid score 0.0), but allergenicity was suggested (hybrid score 0.36), despite the absence of predicted epitopes.PASTA 2.0 analysis identified four N-terminal residues prone to parallel aggregation.The full peptide sequence was predicted as a potential B-cell epitope (score >0.15), and the N-terminal exhibited higher antigenicity (>1.077).However, NetMHCIIpan -4.1 analysis did not predict any strong or weak binding to the most common major histocompatibility complex class II DRB, DQ, and DP alleles. Conclusion:These findings suggest that N-terminal modifications could be beneficial, as they may reduce aggregation and immunogenic potential of the sequence.The safety of the seven most hydrophobic modified sequences will be further evaluated and compared with the original sequence data.In conclusion, this study will contribute to the development of a pipeline for designing nanoparticle formulations to deliver immunobiologicals.