Early Engineering Approaches to Improve Peptide Developability and Manufacturability

Abstract

Abstract. Downstream success in Pharmaceutical Development requires thoughtful molecule design early in the lifetime of any potential therapeutic. Most therapeutic monoclonal antibodies are quite similar with respect to their developability properties. However, the properties of therapeutic peptides tend to be as diverse as the molecules themselves. Analysis of the primary sequence reveals sites of potential adverse posttranslational modifications including asparagine deamidation, aspartic acid isomerization, methionine, tryptophan, and cysteine oxidation and, potentially, chemical and proteolytic degradation liabilities that can impact the developability and manufacturability of a potential therapeutic peptide. Assessing these liabilities, both biophysically and functionally, early in a molecule's lifetime can drive a more effective path forward in the drug discovery process. In addition to these potential liabilities, more complex peptides that contain multiple disulfide bonds can pose particular challenges with respect to production and manufacturability. Approaches to reducing the disulfide bond complexity of these peptides are often explored with mixed success. Proteolytic degradation is a major contributor to decreased half-life and efficacy. Addressing this aspect of peptide stability early in the discovery process increases downstream success. We will address aspects of peptide sequence analysis, molecule complexity, developability analysis, and manufacturing routes that drive the decision making processes during peptide therapeutic development.

In-solution enrichment identifies peptide inhibitors of protein–protein interactions

“…3c and Supplementary Table 4). Additionally, replacement of Trp with an inert side chain such as hexylalanine may be useful for metabolic stability since Trp is prone to degradation 40 . Phe at position 3 was replaced by the more hydrophobic fluorinated phenylalanines, while Leu at position 10 was replaced by bulkier hydrophobic side chains including cyclobutylalanine ( 37 , 39 , 42 , 51 , and 56 ), cyclohexylalanine ( 41 , 45 , and 49 ), or fluorinated phenylalanines ( 43 , 44 , 47 , 52 – 55 , and 57 ; Supplementary Table 4).…”

The Cystine Knot Is Responsible for the Exceptional Stability of the Insecticidal Spider Toxin ω-Hexatoxin-Hv1a

“…In contrast with native toxin, the linear version of Hv1a was only stable at neutral pH values, with marked degradation under both acidic and alkaline conditions. This instability presumably arises from well-known processes that impact on protein stability such as asparagine deamidation, aspartate isomerisation, and racemisation [ 16 , 17 ]. Interestingly, Hv1a contains an Asn-Gly sequence ( Figure 1 ), which has been shown to be highly susceptible to asparagine deamidation [ 18 ].…”

Methods to Enhance the Metabolic Stability of Peptide-Based PET Radiopharmaceuticals

“…One of the key reasons a peptide sequence can also be susceptible to proteolytic degradation is due to its backbone containing a recognition motif for an endogenous protease [31]. In addition to proteolytic degradation, peptide bonds can also undergo spontaneous degradation under physiological conditions when particularly labile sequence motifs are present [32].…”