Solid‐Phase Aggregation of Proteins under Pharmaceutically Relevant Conditions

• Published in: Journal of pharmaceutical sciences 1994-12, Vol.83 (12), p.1662-1669
• Main Authors: Costantino, Henry R., Langer, Robert, Klibanov, Alexander M.
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Chemical Phenomena; Chemistry, Pharmaceutical - methods; Chemistry, Physical; Drug Stability; General pharmacology; Medical sciences; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Proteins - chemistry
• ISSN: 0022-3549; 1520-6017
• DOI: 10.1002/jps.2600831205

In order to successfully employ proteins as pharmaceuticals, it is essential to understand mechanistically the stability issues relevant to their formulation and delivery. Various deleterious processes may occur in protein formulations, thereby diminishing their therapeutic value. This review focuses upon one aspect of this problem, namely aggregation of solid proteins under pharmaceutically relevant conditions (elevated temperature and water activity). Strategies to pursue such studies are presented with an emphasis on a mechanistic analysis of aggregate formation. Both covalent and noncovalent aggregation pathways have been elucidated. Proteins that contain disulfide bonds as well as free thiol residues may aggregate via thiol–disulfide interchange. For proteins which contain disulfides but not free thiol residues, intermolecular disulfide bonding may still occur when intact disulfides undergo β‐elimination, yielding free thiols which can catalyze disulfide scrambling. Finally, proteins containing no cysteine/cystine residues may aggregate by other covalent pathways or by noncovalent routes. On the basis of these pathways, some rational stabilization strategies have been proposed and verified. Ultimately, application of this knowledge should lead to more stable and effective pharmaceutical protein formulations.