Inhalation is the gold standard for many drugs in respiratory medicine, to match the route to the target location. It consists of delivering a drug directly to the respiratory tract, as an aerosol. A major challenge associated with antibody aerosolization is instability, as they are exposed to a huge air-liquid interface along with shearing and rise in temperature, ultimately leading to aggregation. Aggregates generated during aerosolization should be prevented by selecting the appropriate device and formulation, as they may be associated with a reduced efficacy and immunocytotoxicity. 

Inhaled biologics are an emerging class of therapeutics for respiratory diseases and represent attractive alternatives to systemic antibodies. This presentation focuses on multi-staged developability workflows for Pieris inhalable drugs. A wide range of analytical, biophysical, and physicochemical methods are being used for de-risking CMC-development. Preformulation, stability, and stress studies are proving essential for fast and safe drug development.

In this presentation, I will introduce a universal stabilization approach for ACE2-fusion proteins. The stabilization of the ACE2 domain is achieved by small molecular compounds that bind to the protein with high affinity and inhibit its enzymatic activity. The binding and stabilization mechanisms were studied with a combination of complementary techniques including differential scanning calorimetry, isothermal titration calorimetry, and hydrogen-deuterium exchange mass spectrometry.

Forced degradation of biopharmaceutical proteins is a staple study of the industry and serves both to identify the proteins’ weak points as well as to develop analytical methods to characterize them. The recent molecular model learnings on methionine and tryptophan oxidation, asparagine deamidation, and aspartate isomerization can be integrated directly from the literature into the development workflow, bringing significant benefits, such as:

• Reduction in material requirements
• Significant reduction of experimental workload
• Critical quality attribute assessment - e.g., calculating the impact of posttranslational modifications on bioactivity?

• Is SC administration more immunogenic than other routes?
• What are the contributing attributes such as aggregate content, oxidized species, injection rate, protein concentration, formulation and excipients, impurities) and how can these attributes be assessed in a patient-centric manner? 
• What are appropriate models (in vitro, in vivo) that can be used to determine immunogenicity, and can these form the basis for specifications? 
• How immunogenic are new modalities that deviate from standard monoclonal antibody platforms??

• What are the current challenges and limitations in mRNA delivery, and how can they be overcome?
• What are the key considerations when designing delivery systems for targeted mRNA delivery? How can we optimize the delivery efficiency and specificity?
• What are some of the most promising applications of targeted mRNA delivery in medicine (e.g. in vivo CAR-T production)? What are the benefits compared to other strategies?

Multi-dose formulations contain preservatives to prevent growth of microorganisms during the in-use timeframe. The destabilization of proteins by preservatives is a major challenge in the development of multi-dose biologics. We have evaluated Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS) to measure changes in structure and flexibility of model antibody in the presence of three preservatives. The results identified a common hot spot for preservative interaction and showed a pattern of altered antibody backbone flexibility in the presence of preservatives. HDX-MS is a promising technique to integrate into early workflows to identify candidate antibodies with amenable properties for multi-dose formulations.

Due to the high doses required in most treatments and the small volume that is admitted for subcutaneous administration, it is necessary to obtain formulations of antibodies at high concentrations. A staged screening methodology was used to determine the best formulation for the nimotuzumab. This mAb is an anti-EGFR antibody that is used in the treatment of some different tumours. The nimotuzumab antibody was formulated in high concentrations (> 150 mg/mL). Physicochemical and biological characterization and stability were determined where no significant changes were observed. Loss in biological activity was observed. Additionally, the antitumour effect and PK properties were analyzed.

IV admixture compatibility and in-use stability are critical components in ensuring patient safety and product efficacy. Case studies on antibody drug products  will be presented, including preventing low dose adsorption and using proper ancillaries for dose preparation to ensure dose accuracy, as well as defining in-use time and minimizing IRRs to improve safety. The use of CSTD for dose preparation and administration will also be discussed.

This talk delves into the surfactant-related stability challenges encountered in the biopharmaceutical industry. In particular, it will cover contributing CMC factors leading to free fatty acid particles derived from polysorbate formulations, and protein-silicone oil (PDMS) particles observed in poloxamer 188 (P188) formulations. This talk will also discuss the general risk profiles and mitigation approaches associated with different routes of administration (e.g., IV, SC, IVT).