Enhancement of The Stability of Human Growth Hormone by Using Tris(hydroxymethyl)aminomethane: Molecular Docking and Experimental Analysis

Mirzaei, Siyavash; Mobedi, Hamid; Gourabi, Hamid; Sanati, Mohammad Hosein; Khezli, Sakine; Omidian, Hamid; Ighaeie, Masoume

doi:10.22074/cellj.2021.6903

Enhancement of The Stability of Human Growth Hormone by Using Tris(hydroxymethyl)aminomethane: Molecular Docking and Experimental Analysis

Document Type : Original Article

Authors

¹ .Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

² .Department of Novel Drug Delivery Systems, Iran Polymer and Petrochemical Institute, Tehran, Iran

³ .Medical Genetics Department, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran

⁴ 4.Department of Pharmaceutical Sciences, Nova Southeastern University, Pharmaceutical Sciences, Davie, Florida, USA

⁵ 5.Computational Nano Physical Chemistry Laboratory, Department of Chemistry, Azerbaijan Shahid Madani University, Tabriz, Iran

10.22074/cellj.2021.6903

Abstract

Objective
It is so difficult to formulate human growth hormone (hGH) in a solution with high stability and new drug delivery system (NDDs) due to physiochemical instability. The purpose of this study was to investigate the possibility of using Tris as a hGH stabilizer.
Materials and Methods
In this experimental study, the role of tris(hydroxymethyl)aminomethane (Tris) was evaluated as a hGH stabilizing agent in phosphate buffer, as a practical aqueous solution and a media to release NDDs. High- performance liquid chromatography (HPLC) and enzyme-linked immune sorbent assay (ELISA) were applied to investigate the stability of hGH in solutions and dynamic light scattering (DLS) was used to measure the effect of Tris on the hydrodynamic size of hGH in aqueous solutions. Ultra violet (UV) spectrophotometry was used to check the hGH spectrum. In computational study, formation of ligand-protein complex of the Tris-hGH, and the intermolecular interactions between Tris and hGH were studied by molecular docking modeling.
Results
The results demonstrated that Tris at the optimum concentration, increases hGH stability in aqueous solutions. Also, molecular docking modeling confirmed that amino acid residues such as tyrosine (Tyr), proline (Pro), glutamic acid (Glu), aspartic acid (Asp), leucine (Leu), and phenylalanine (Phe) in hGH structure, were linked with Tris as a ligand.
Conclusion
It seems that interactions between hGH and Tris are the most important reason for increment of the physicochemical stability of hGH in aqueous solutions containing Tris.

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