(281g) Fundamental Motions: The Key to Accelerated Lyophilized Formulation Development

Protein-based therapeutic drugs have served an important role in the treatment of several diseases and conditions, including influenza and certain cancers. Preservation of the proteins within such drugs is critical for patient safety and treatment efficacy. Lyophilization (freeze-drying) has been shown as an effective technique for enhancing the longevity of proteins, however, degradation is still imminent in many cases. While longer shelf-life is obviously advantageous, the result is long degradation studies to determine what formulations are most suitable for the stability of the protein within the drug product. In this work, I will demonstrate how probing “fundamental motions” in a lyophilized system can provide the insight required to accelerate lyophilized protein-based drug formulation development.

A 2012 publication by Cicerone¹ provided evidence for a relationship between the degradation rates of several proteins and the dynamics of the excipients that the proteins were lyophilized in. There are several works by others that investigate system dynamics through viscosity and glass-transition temperature, but to this end, show no clear correlation with stability. The publication by Cicerone, however, looked at more fundamental motions of the excipients: individual molecules hopping over each other. This data was acquired with neutron scattering, a technique less accessible to academia and industry, so the present work explores optical techniques to acquire similar data.

In this study, the long-term stability of lyophilized alcohol dehydrogenase (ADH), horseradish peroxidase (HRP), and IgG1 are characterized with enzymatic assays and size-exclusion chromatography. This data is then compared to the optical techniques which provide information regarding quantity of fundamental motions of the system (less motion means more stable protein). As these optical methods take on the order of minutes, with this work, we approach a promising method to reduce lyophilized protein formulation development times from a year to minutes.

1. Cicerone et al., Soft Matter, 8 (2012)