brij-58 and Hemolysis

Polysorbates (PSs) are common protein stabilizers used in biotherapeutic formulations. However, PSs are heterogeneous and unstable in liquid protein formulations [1,2]. The purpose of this work is to explore possible alternatives for polysorbate replacements that demonstrate superior protein protection, superior self-stability, low toxicity, and wide applicability. For this purpose, 8 non-ionic surfactants that have not yet been used as excipients in marketed biotherapeutic products were investigated with PS20/80 as the benchmark. Compared with PS20/80, Brij-58 showed better protein protection ability in the mAb1 formulation under forced degradation conditions when examined by visual inspection, SEC, and dynamic lighting scanning. Additionally, Brij-58 has a better inherent stability than PS20/80 in the protein formulation when detected by UPLC-CAD. Moreover, Brij-58 is an inert excipient that does not affect protein bioactivity and conformation. In addition, the LD50 and hemolysis concentration of Brij-58 were determined, which is relatively safe when used as a parenteral injection. Furthermore, Brij-58 was also an effective protein stabilizer for the other two antibody products (IgG4 subtype and bispecific antibody) in the shaking study. In summary, Brij-58 stands out as a promising PS replacement in biotherapeutic formulations with a safe, stable and effective protein-protection profile among candidate surfactants.

Topics: Administration, Intravenous; Animals; Antibodies, Bispecific; Antibodies, Monoclonal; Biological Products; Cetomacrogol; Chemistry, Pharmaceutical; Drug Compounding; Drug Stability; Excipients; Female; HEK293 Cells; Hemolysis; Humans; Immunoglobulin G; Lethal Dose 50; Male; Mice; Polysorbates; Protein Stability; Rabbits; Surface-Active Agents; Toxicity Tests, Acute

The mechanism of cetomacrogol 1000-induced hemolysis was investigated. Previous conclusions that peroxides are involved in the hemolytic process were confirmed. The possibility that hydrogen peroxide, superoxide, hydroxyl radical, or singlet oxygen, which are known to induce hemolysis, are involved in cetomacrogol 1000-induced hemolysis was tested by using specific inhibitors and inactivators. The hydroxyl radical (OH.) was shown to be the only apparent oxygen species involved in cetomacrogol 1000-induced hemolysis. Its contribution to the hemolytic potency of the surfactant is approximately 30%.

Topics: Animals; Catalase; Cetomacrogol; Free Radicals; Hemolysis; Histidine; Hydroxides; Hydroxyl Radical; In Vitro Techniques; Light; Mannitol; Polyethylene Glycols; Rats; Saponins; Superoxide Dismutase; Thiourea

The hemolytic properties of nonionic surfactants of the series CH3(CH2)15-17-O-(CH2CH2O)nCH2CH2OH were investigated and compared to those of saponins, sapogenins and H2O2. Antioxidants and anaerobic conditions were shown to inhibit the hemolysis, while glycyrrhizin was found to enhance it. Similar effects were obtained for H2O2 hemolysis, but not for saponin and sapogenin hemolysis. It is proposed that peroxides and free radicals are mainly responsible for the polyoxyethylene derived surfactants induced hemolysis.

Topics: Animals; Cetomacrogol; Glycyrrhetinic Acid; Glycyrrhizic Acid; Hemolysin Proteins; Hemolysis; Peroxides; Polyethylene Glycols; Rats; Sapogenins; Saponins; Surface-Active Agents