ReviewsPolysorbates 20 and 80 Used in the Formulation of Protein Biotherapeutics: Structure and Degradation Pathways
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INTRODUCTION
The proper formulation of recombinant proteins for pharmaceutical products is vital to their long-term stability and reproducible activity. To maintain biological activity, proteins generally must be maintained in a specific, three-dimensional conformation. This conformation is only marginally stable, and thus relatively minor perturbing forces can disrupt protein structure causing loss of biological activity or an immunological response. Such perturbations are commonly encountered as proteins
POLYSORBATE STRUCTURE
The polysorbates are amphipathic, nonionic surfactants composed of fatty acid esters of polyoxyethylene sorbitan.21 Polysorbate 20 (polyoxyethylene sorbitan monolaurate) and Polysorbate 80 (polyoxyethylene sorbitan monooleate) are the most common polysorbates currently used in formulation of protein biopharmaceuticals.22 Their structures are shown in Figure 1. The structures shown here are those of the chemically homogenous polysorbates. Solutions of polysorbate sold by manufacturers and
SOLUTION AND SURFACE ACTIVE PROPERTIES OF POLYSORBATE
Because of their dual hydrophobic/hydrophilic nature, surfactants in solution tend to orient themselves so that the exposure of the hydrophobic portion of the surfactant to the aqueous solution is minimized.20 In systems containing air/water interfaces, surfactants will tend to accumulate at these interfaces, forming a surface layer of surfactant oriented in such a fashion that only their hydrophilic ends are exposed to water.20, 27, 28 Such orientation and hydrophobic surface adsorption can
CHEMICAL STABILITY OF POLYSORBATES
Chemical stability of the polysorbates is another important consideration for their use. The polysorbates are notorious for undergoing autooxidation34, 73., 74., 75., 76., 77., 78., 79. and cleavage at the ethylene oxide subunits34, 74 as well as hydrolysis of the fatty acid ester bond.34, 80 A generalized reaction scheme depicting these processes is shown in Figure 6. Autooxidation of the ethylene oxide results in hydroperoxide formation, side-chain cleavage and eventually formation of short
CONCLUDING REMARKS
Polysorbates are present in a large number of biopharmaceutical drugs listed in the 2006 Physicians Desk Reference. The concentrations used in the formulations range from 0.0003% (w/v) to 0.3% (w/v). Over the past decade the quality of polysorbate solutions has increased dramatically such that many manufacturers now offer highly purified, low peroxide, and low acid content solutions. While the solutions are initially low in reactive oxygen species care must still be used when storing drug
ACKNOWLEDGEMENTS
The author would like to thank Dr. Sungae Park for the information from the European Pharmacopoeia, Arnold McAuley, and Hyo Jin Le for the surface tension measurements and Dr. Darren Reid, Dr. David Brems, and Dr. Richard Remmele for many useful discussions and critical reading of the manuscript.
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