Rational approach for design and evaluation of anti-aggregation agents for protein stabilization: A case study of trehalose phenylalaninate
| Autor: | Smita S. Kale, Krishnacharya G. Akamanchi |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Pharmaceutical Science Excipient Protein aggregation 010402 general chemistry 01 natural sciences Excipients 03 medical and health sciences Molecular dynamics chemistry.chemical_compound medicine Bovine serum albumin Quenching (fluorescence) biology Protein Stability Anti aggregation Temperature Trehalose Serum Albumin Bovine 0104 chemical sciences Molecular Docking Simulation 030104 developmental biology chemistry Biochemistry Biophysics biology.protein Protein stabilization medicine.drug |
| Zdroj: | International journal of pharmaceutics. 524(1-2) |
| ISSN: | 1873-3476 |
| Popis: | The present work introduces new anti-aggregation agent (AAA) derived through our new approach for design and evaluation of anti-aggregation agent as a multi-purpose excipient to combat protein aggregation. Therapeutic proteins undergo aggregation due to even minor changes in environmental conditions like temperature, pH, shear and stress. Excipients play a vital role in prevention of aggregation. To stabilize a protein formulation different classes of excipients are used in combination after carefully selecting through laborious and time consuming trial and error experiments. To resolve these concerns, we have developed a rational approach based on molecular docking analysis and have designed, synthesized AAAs, and validated the approach by experimental studies. Trehalose phenylalaninate (TPA) has been synthesized and evaluated for stabilization of Bovine serum albumin (BSA). TPA was found to be non-toxic with a LC50 of >80 μg/ml. BSA solutions with and without TPA were subjected to thermal and agitation stress and aggregation was monitored using sophisticated analytical techniques. The helical structure of BSA was completely retained in stressed samples at 0.1% concentration of TPA. SEC-HPLC clearly demonstrated the absence of aggregates in presence of TPA. Although aggregation was not seen in fluorescence spectra but quenching due to TPA was evident. Moreover, molecular dynamics study on BSA-TPA complex showed lower RMSD. |
| Databáze: | OpenAIRE |
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