Stability and Structural Analysis of hGH and Infliximab for Controlled Release Depots and Biosimilars.

controlled release;biosimilar;protein therapeutics;Pharmacy and Pharmacology;Health Sciences;Pharmaceutical Sciences

Pisupati, KarthikRodriguez-Hornedo, Nair ;

University of Michigan

关键词: controlled release;    biosimilar;    protein therapeutics;    Pharmacy and Pharmacology;    Health Sciences;    Pharmaceutical Sciences;   

Others  :  https://deepblue.lib.umich.edu/bitstream/handle/2027.42/120777/pkarthik_1.pdf?sequence=1&isAllowed=y

瑞士|英语

来源: The Illinois Digital Environment for Access to Learning and Scholarship

【 摘 要 】

Therapeutic proteins have revolutionized modern medicine and are a rapidly growing drug class. However, these complex entities suffer from high cost, instability, and the necessity for frequent injections. This thesis advanced biosimilar and controlled release formulation development to address these shortcomings by testing 2 hypotheses: (a) aqueous remote loading of Zn-binding therapeutic proteins in poly(lactic-co-glycolic acid) (PLGA) microspheres can be accomplished with high efficiency and protein loading for later controlled release of stable protein; and (b) state-of-the-art analytical methods can rapidly identify key structural similarities and differences between competing antibody products before and after stressing. PLGA microsphere depots with interconnecting pore networks and poorly soluble Zn bases (e.g. ZnCO3) were incubated with mildly acidic solutions of Zn-binding human growth hormone (hGH). hGH precipitated in the polymer pores with Zn2+ and was encapsulated after healing/closing the PLGA pores with elevated temperature (43°C). Zn:hGH complexes formed efficiently at a 10:1 mol ratio and protein stabilization was afforded by the presence of disaccharides. Under similar conditions, Zn-containing PLGA microspheres were successfully loaded with hGH up to 4.3 % w/w with 86% efficiency with stable protein release for over 4 weeks. The protein loading was controlled by adjusting loading time, Zn content, and microsphere size. The release buffer affected whether hGH release obeyed first order (in phosphate buffer) or near zero-order (in HEPES) kinetics.Additionally for biosimilars, an originator, Remicade (infliximab), and its copy, Remsima, were assessed via two strategies: state of the art mass spectrometery driven analysis, and analytical characterization post stressing. Remicade and Remsima displayed highly similar primary and higher-order structures and primary chemical modifications. Remicade, however, possessed higher levels of a-fucosylated glycans and tighter Fc-gamma receptor binding, suggesting a possible difference in efficacy in inflammatory bowel disease indications. Despite small initial differences, upon stressing, both products formed the same by-products at similar rates, further verifying the structural biosmilarity of these products. Ultimately this work demonstrates an encapsulation and analytical framework, which can be applied towards future PLGA and biosimilar formulation development, in order to improve the safety and efficacy of monoclonal antibody and Zn-binding protein therapeutics.