Rational Design of Biobetters with Enhanced Stability

• Published in: Journal of pharmaceutical sciences 2015-08, Vol.104 (8), p.2433-2440
• Main Authors: Courtois, Fabienne, Schneider, Curtiss P., Agrawal, Neeraj J., Trout, Bernhardt L.
• Format: Article
• Language: English
• Subjects: Amino Acid Substitution; Animals; Antibodies, Monoclonal, Humanized - adverse effects; Antibodies, Monoclonal, Humanized - chemistry; Antibodies, Monoclonal, Humanized - metabolism; Antibodies, Monoclonal, Humanized - pharmacology; Antibody Affinity; Antigens, CD20 - chemistry; Antigens, CD20 - metabolism; Antineoplastic Agents - adverse effects; Antineoplastic Agents - chemistry; Antineoplastic Agents - metabolism; Antineoplastic Agents - pharmacology; B-Lymphocytes - drug effects; B-Lymphocytes - metabolism; biobetter; Burkitt Lymphoma - drug therapy; Burkitt Lymphoma - metabolism; Cell Line, Tumor; Drug Design; Drug Stability; Epitopes - metabolism; Humans; Immunoglobulin Fab Fragments - chemistry; Immunoglobulin Fab Fragments - genetics; Immunoglobulin Fab Fragments - metabolism; Immunoglobulin Fab Fragments - pharmacology; Models, Molecular; molecular modeling; monoclonal antibody; Mutagenesis, Site-Directed; Mutant Proteins - adverse effects; Mutant Proteins - chemistry; Mutant Proteins - metabolism; Mutant Proteins - pharmacology; mutation; Protein Aggregates; protein aggregation; Protein Engineering; Protein Stability; Recombinant Proteins - adverse effects; Recombinant Proteins - chemistry; Recombinant Proteins - metabolism; Recombinant Proteins - pharmacology; rituximab; Rituximab - chemistry; Rituximab - genetics; Rituximab - metabolism; Rituximab - pharmacology; SAP; stabilization
• ISSN: 0022-3549; 1520-6017
• DOI: 10.1002/jps.24520

Biotherapeutics are the fastest growing class of pharmaceutical with a rapidly evolving market facing the rise of biosimilar and biobetter products. In contrast to a biosimilar, which is derived from the same gene sequence as the innovator product, a biobetter has enhanced properties, such as enhanced efficacy or reduced immunogenicity. Little work has been carried out so far to increase the intrinsic stability of biotherapeutics via sequence changes, even though, aggregation, the primary degradation pathway of proteins, leads to issues ranging from manufacturing failure to immunological response and to loss of therapeutic activity. Using our spatial aggregation propensity tool as a first step to a rational design approach to identify aggregation-prone regions, biobetters of rituximab have been produced with enhanced stability by introducing site-specific mutations. Significant stabilization against aggregation was achieved for rituximab with no decrease in its binding affinity to the antigen.