Improving Prediction of Free Fatty Acid Particle Formation in Biopharmaceutical Drug Products: Incorporating Ester Distribution during Polysorbate 20 Degradation.

Abstract

Polysorbate 20 (PS20) is a commonly used surfactant in biopharmaceutical formulations. It is a heterogeneous surfactant containing a distribution of fatty acid esters, which are subject to hydrolytic degradation, generating free fatty acids (FFAs). The FFAs can form visible or subvisible particles in drug product on stability. A previous FFA solubility model, developed by our group, predicts solubility limits for the three most prevalent FFA degradation products of PS20: lauric, myristic, and palmitic acid. The model takes into account two formulation parameters, pH and PS20 concentration, and their effect on FFA solubility. This work identifies a third parameter that has an impact on FFA solubility: PS20 ester distribution. When PS20 is hydrolytically degraded, the ester distribution of the remaining surfactant changes on stability. Ester distribution is known to influence the critical micelle concentration (CMC) of PS20 such that the monoesters have a much higher CMC compared to the higher-order esters (HOE). We hypothesize that as PS20 degrades, the CMC changes, affecting the proportion of PS20 that is present in micelles and capable of sequestering and solubilizing FFAs in these micelles. Here, PS20 was separated into monoester, HOE, and polyol fractions. The monoester and HOE fractions were mixed together to generate the mock degradation profiles of hydrolytically degraded PS20. FFA solubility was measured as a function of the concentration of these mock-degraded (MD) PS20s. The results indicate that ester distribution does have an impact on FFA solubility, especially at higher MD PS20 concentrations. HOEs solubilize up to 30 μg/mL more lauric acid than an equivalent amount of monoesters at a MD PS20 level of 0.06% w/v. With the addition of % HOE peak area fraction as a third parameter representing the ester distribution of PS20, the refined FFA solubility model more accurately predicts FFA solubility in protein formulations at 5 °C. The refined model suggests that drug products containing trace levels of host cell proteins (HCPs) that preferentially degrade HOEs of PS20 are at a higher risk of particle formation.