Proteolysis and Oxidation of Therapeutic Proteins After Intradermal or Subcutaneous Administration

Abstract

Keywords:intradermal subcutaneous therapeutic proteins PBPK modeling reactive oxygen species proteolysis oxidation lymphatic system a b s t r a c tThe intradermal (ID) and subcutaneous (SC) routes are commonly used for therapeutic proteins (TPs) and vaccines; however, the bioavailability of TPs is typically less than small molecule drugs given via the same routes. Proteolytic enzymes in the dermal, SC, and lymphatic tissues may be responsible for the loss of TPs. In addition, the TPs may be exposed to reactive oxygen species generated in the SC tissue and the lymphatic system in response to injection-related trauma and impurities within the formulation. The reactive oxygen species can oxidize TPs to alter their efficacy and immunogenicity potential. Mechanistic understandings of the dominant proteolysis and oxidative routes are useful in the drug discovery process, formulation development, and to assess the potential for immunogenicity and altered pharmacokinetics (PK). Furthermore, in vitro tools representing the ID or SC and lymphatic system can be used to evaluate the extent of proteolysis of the TPs after the injection and before systemic entry. The in vitro clearance data may be included in physiologically based pharmacokinetic models for improved PK predictions. In this review, we have summarized various physiological factors responsible for proteolysis and oxidation of TPs after ID and SC administration.

Decoding the proteome of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) for cell-penetrating peptides involved in pathogenesis or applicable as drug delivery vectors

“…A protease or peptidase is an enzyme that hydrolyzes proteins and peptides to polypeptides or single amino acids. Proteases are categorized into four families as follows: 1) aspartic proteases such as pepsins, 2) cysteine proteases namely caspases, 3) metalloproteases like matrix metallopeptidase, and 4) serine proteases such as plasmin and thrombin ( Varkhede et al, 2020 ). Inherent susceptibility of peptides to protease degradation and their further inactivation would significantly prevent their development as therapeutic agents ( Heard et al, 2013 ).…”

Stability of monoclonal antibodies after simulated subcutaneous administration

“…Although the basal proteolytic activity in the SC tissue is low, administration of a therapeutic protein can enhance enzymatic activity. 43 This is particularly important when assessing enzymatically-driven degradation events such as fragmentation. Cells are fundamental modulators from a PK/PD perspective (e.g., potency), however, previous studies have shown limited stability of fluids due to cell lysis in vitro 44 and potential clogging of analytical instruments.…”

“…45 With an increasing complexity of fluids (e.g., neat biological fluid or spiking endogenous proteins to fluids), analytical challenges arise, which can cause matrix interference. 43 More complex in vitro models, require additional analytical procedures such as fluorescence labeling or purification (e.g., biotinylation) of the protein of interest to enable detectability. However, altering the protein of interest may create measurement artifacts and limit the possibility to investigate certain degradation products.…”

Functionalized Particles Designed for Targeted Delivery

“…Degradation of proteins occurs mainly by enzymatic pathways, regardless of whether in vitro [ 148 ] or in vivo [ 149 , 150 ]. Orally administered protein carriers are degraded by digestive enzymes [ 149 ] whereas those introduced directly into the bloodstream enter cells by endocytosis and undergo lysosomal degradation [ 142 ].…”