Trehalose Monooleate: A Potential Antiaggregation Agent for Stabilization of Proteins
| Autor: | Smita S. Kale, Krishnacharya G. Akamanchi |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Pharmaceutical Science Excipient 02 engineering and technology Protein aggregation Excipients Surface-Active Agents 03 medical and health sciences chemistry.chemical_compound Drug Discovery medicine Animals Bovine serum albumin chemistry.chemical_classification Chromatography biology Protein Stability Chemistry Trehalose Serum Albumin Bovine Polymer 021001 nanoscience & nanotechnology Amino acid Polysorbates Spectrometry Fluorescence 030104 developmental biology Docking (molecular) biology.protein Molecular Medicine Cattle Protein Multimerization 0210 nano-technology medicine.drug |
| Zdroj: | Molecular Pharmaceutics. 13:4082-4093 |
| ISSN: | 1543-8392 1543-8384 |
| DOI: | 10.1021/acs.molpharmaceut.6b00686 |
| Popis: | Protein aggregation is a major problem of therapeutic proteins because aggregation decreases their therapeutic activity and shelf life and induces immunogenicity. Stabilization against aggregation is commonly attained by addition of different excipients like sugars, surfactants, buffers, salts, amino acids, polymers, etc. Generally these excipients are required in combination for stabilization. Sugars are required at a higher concentration, and commonly used surfactants like polysorbates have shortcomings due to oxidative degradation. With a view to have a multipurpose excipient to be effective at a lower concentration, we designed antiaggregation agents (AAAs) that would encompass the functionalities of two or more conventional excipients and would curtail the number of excipients to be added for stabilization. Our first designed AAA, trehalose monooleate (TMO), is a sugar-fatty acid derivative. It has been evaluated in silico by docking on aggregation prone regions of model protein bovine serum albumin (BSA), and experimentally its effectiveness has been validated as stabilizer against agitation and thermal stress. TMO has a lower CMC of 6 mg/L, is nonhemolytic, and was found to be nontoxic by sulforhodamine B (SRB) colorimetric assay in Human Hepatoma Cell Line (Hep-G2) using adriamycin as positive contol. Various spectroscopic and separation analytical techniques were employed to monitor the aggregation profile of BSA in presence and absence of TMO. CD spectroscopy showed complete retention of helical structure at concentration as low as 0.05% of TMO, while fluorescence spectroscopy provided vital insights into conformational stability rendered by TMO. Native-PAGE and SEC-HPLC studies demonstrated absence of aggregates. Molecular dynamics study on BSA-TMO docked complex further substantiated the stabilization effect. Overall, it can be said that TMO has good antiaggregation property. The present work is a preliminary attempt toward understanding protein excipient interactions and chemistry to provide rational basis for designing a single excipient for stabilization of protein formulations. |
| Databáze: | OpenAIRE |
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