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In Silico design of AVP (4–5) peptide and synthesis, characterization and in vitro activity of chitosan nanoparticles

  • Serda Kecel-GunduzEmail author
  • Yasemin Budama-Kilinc
  • Rabia Cakir-Koc
  • Tolga Zorlu
  • Bilge Bicak
  • Yagmur Kokcu
  • Aysen E. Ozel
  • Sevim Akyuz
Research article

Abstract

Background

Arginine-vasopressin (AVP) is a neuropeptide and provides learning and memory modulation. The AVP (4–5) dipeptide corresponds to the N-terminal fragment of the major vasopressin metabolite AVP (4–9), has a neuroprotective effect and used in the treatment of Alzheimer’s and Parkinson’s disease.

Methods

The main objective of the present study is to evaluate the molecular mechanism of AVP (4–5) dipeptide and to develop and synthesize chitosan nanoparticle formulation using modified version of ionic gelation method, to increase drug effectiveness. For peptide loaded chitosan nanoparticles, the synthesized experiment medium was simulated for the first time by molecular dynamics method and used to determine the stability of the peptide, and the binding mechanism to protein (HSP70) was also investigated by molecular docking calculations. A potential pharmacologically features of the peptide was also characterized by ADME (Absorption, Distribution, Metabolism and Excretion) analysis. The characterization, in vitro release study, encapsulation efficiency and loading capacity of the peptide loaded chitosan nanoparticles (CS NPs) were performed by Dynamic Light Scattering (DLS), UV–vis absorption (UV), Scanning Electron Microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy techniques. Additionally, in vitro cytotoxicity of the peptide on human neuroblastoma cells (SH-SY5Y) was examined with XTT assay and the statistical analysis was evaluated.

Results

The results showed that; hydrodynamic size, zeta potential and polydispersity index (PdI) of the peptide-loaded CS NPs were 167.6 nm, +13.2 mV, and 0.211, respectively. In vitro release study of the peptide-loaded CS NPs showed that 17.23% of the AVP (4–5)-NH2 peptide was released in the first day, while 61.13% of AVP (4–5)-NH2 peptide was released in the end of the 10th day. The encapsulation efficiency and loading capacity were 99% and 10%, respectively. According to the obtained results from XTT assay, toxicity on SHSY-5Y cells in the concentration from 0.01 μg/μL to 30 μg/μL were evaluated and no toxicity was observed. Also, neuroprotective effect was showed against H2O2 treatment.

Conclusion

The experimental medium of peptide-loaded chitosan nanoparticles was created for the first time with in silico system and the stability of the peptide in this medium was carried out by molecular dynamics studies. The binding sites of the peptide with the HSP70 protein were determined by molecular docking analysis. The size and morphology of the prepared NPs capable of crossing the blood-brain barrier (BBB) were monitored using DLS and SEM analyses, and the encapsulation efficiency and loading capacity were successfully performed with UV Analysis. In vitro release studies and in vitro cytotoxicity analysis on SHSY-5Y cell lines of the peptide were conducted for the first time.

Grapical abstract

Keywords

AVP (4–5) Hsp70 Drug delivery Nanoparticle Chitosan Parkinson MD 

Abbreviations

AVP

Arginine-vaspressin

CS NPs

Chitosan nanoparticles

AD

Alzheimer’s disease

PD

Parkinson’s disease

NGF

Nerve growth factor

HSP

Heat shock proteins

MD

Molecular Dynamics

NVT

Number of particles, Volume, and Temperature

NPT

Number of particles, Pressure, and Temperature

Rg

Gyration

RMSD

Root mean square deviation

VMD

Visual Molecular Dynamics

ADME

Absorption, Distribution, Metabolism and Excretion

TED

Total energy distribution

PdI

Polydispersity index

FT-IR

Fourier Transform Infrared

SEM

Scanning Electron Microscopy

DLS

Dynamic Light Scattering

EE

Encapsulation Efficiency

LC

Loading Capacity

PBS

Phosphate Buffered Saline

XTT

sodium 3,3′-[1(phenylamino)carbonyl]-3,4-tetrazolium]-3is(4-methoxy-6-nitro) benzene sulfonic acid hydrate

DMSO

Dimethyl Sulfoxide

Notes

Acknowledgements

Authors are also very thankful to Rita Podzuna for allowing using the docking program with Schrödinger’s Small-Molecule Drug Discovery Suite. In this study, the infrastructure of Applied Nanotechnology and Antibody Production Laboratory established with TUBITAK support (project numbers: 115S132 and 117S097) was used. Authors would thank to TUBITAK for their support.

Availability of data and materials

Data sharing not applicable to this article as no datasets were generated or analysed during the current study.

Authors’ contributions

SG: Participated in the design of the study, carried out the FTIR, band component analysis study, molecular docking, and molecular dynamic simulation and drafted the manuscript. YBK and TZ: Participated in the design of the experimental study, (synthesize and characterize nanoparticles) drafted the manuscript. RK: Participated in the design of the experimental study (cytotoxicity studies) drafted the manuscript. BB and YK: Participated in the design of the molecular docking and molecular dynamic simulation. AO and SA: Responsible for the study design and gave final approval of the version to be published. All authors read and approved the final manuscript and provide financial and administrative support.

Funding

This study was supported by the Research funds of Istanbul University [ONAP-2423].

Compliance with ethical standards

Conflict of interests

On behalf of all authors, the corresponding author states that there is no conflict of interest.

Consent for publication

Not applicable.

Ethics approval and consent to participate

Not applicable.

Supplementary material

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Copyright information

© Springer Nature Switzerland AG 2020

Authors and Affiliations

  1. 1.Physics Department, Faculty of ScienceIstanbul UniversityIstanbulTurkey
  2. 2.Department of Bioengineering, Faculty of Chemical and Metallurgical EngineeringYildiz Technical UniversityIstanbulTurkey
  3. 3.Graduate School of Natural and Applied ScienceYildiz Technical UniversityIstanbulTurkey
  4. 4.Department of Physical Chemistry and EMaSUniversitat Rovira i VirgiliTarragonaSpain
  5. 5.Institute of Graduate Studies in SciencesIstanbul UniversityIstanbulTurkey
  6. 6.Physics Department, Science and Letters FacultyIstanbul Kultur UniversityIstanbulTurkey

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