Innovative Pharmaceutical Stabilization and Formulation Techniques for Protein Drugs and Their Influence on Sequence and Structure

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The formulation and administration of pharmacologically relevant proteins has hitherto been limited to the parenteral administration of solutions or liquid dosage forms. This is often due to a lack of stability or lack of tissue specificity of the proteins. In addition, post-translational modifications must be retained in a formulation as they are often critical for protein activity. Apart from bioavailability, these aspects also need to be considered when evaluating alternative formulations and modes of administration. In particular, solids for peroral, nasal or parenteral administration would significantly expand or improve the therapeutic use and the compliance of protein-based drugs. In addition to stability, the evaluation of recipe and process parameters on the microscale and their scalability in the laboratory and pilot scale is of crucial importance, as due to the cost of proteins conventional developments on a gram or kilogram scale are not financially feasible. The aim of this project proposal is the establishment of scalable formulation technologies (melt extrusion, fluidized bed processes and spray freeze drying) for the production of solid dosage forms. In the process, the melt extrusion and spray freezing processes we have established will be transferred to proteins. The stabilization of the proteins should take place by embedding in eroding or dissolving polymer matrices. In addition, it will be investigated by the formulation of highly porous lyophilizate particles of the substances, whether the rapid freezing step of the spray freeze drying can give decisive advantages for protein stability. Various peptides/proteins (for example tridegin, insulin, conopeptides such as ziconotide) are available as models, so that from low-dose to high-dose and from oral targets to implants the entire range of formulation finding can be represented. Statements regarding stability, activity and in vitro release kinetics must be made on each scale. The substances are multiple disulfide-bridged molecules whose correct folding is crucial for their activity. Obtaining the correct disulfide bonding is essential for the formulation of proteins, but also for the development of appropriate standards for their analysis. Chromatography-coupled mass spectrometric methods are usually used to analyze such sequences, while the method of N-terminal protein sequencing according to Edman was neglected in the past decades. In view of the now known limits of mass spectrometric protein analysis, a combination of both methods seems to make sense for a comprehensive protein analysis and will provide information on the stability of the formulated substances in the context of the present application.