Under the Action, E40’s efficacy has been confirmed and its safety demonstrated in regulatory-compliant in vivo studies.
With regards to safety:
- A sensitisation and irritation test was completed, which demonstrated a lack of skin sensitisation potential and identified E40 as a non-sensitising agent; and
- Acute and sub-acute repeated dose toxicity studies established no-observed adverse effect level (NOAEL) that supports calculation of a human equivalent dose (HED) adequate to enable progression to clinical evaluation. The derived HED significantly exceeds the standard safety factors typically applied for selection of first-in-human starting doses.
Collectively, these studies not only confirmed the absence of E40-related toxicity but also validated safe handling conditions. With regards to efficacy, work under the Action confirmed previously generated in vitro efficacy data in a relevant in vivo model of CeD, strengthening translational confidence and de-risking subsequent clinical trials.
Specifically, using the in vivo model of CeD, we demonstrated:
- E40’s proteolytic action on gliadin in vivo;
- A highly effective gluten degradation in the stomach;
- Complete detoxification of the alpha-gliadin derived 33-mer, commonly regarded as the most immunogenic gluten peptide, and significant reduction of other peptides carrying immunogenic sequences specific to gamma-gliadin, confirming previous in vitro findings; and
- Increased activation of cellular antioxidant response.
Collectively, these findings identify E40 to be a highly efficient and effective gluten degrading enzyme, and an ideal candidate for the prevention of unintentional symptomatic gluten exposure.
From a manufacturing standpoint, significant progress has been made in the manufacturing process of E40 with E40 representative of the commercial form now reliably produced at industrial scale. Further, the manufacturing process has been replicated at a secondary site providing supply chain resilience options. Collectively, activities taken under the Action have resulted in a substantial production yield increase and the establishment of a robust, GMP-compliant, high-scale manufacturing process.
Having achieved an economic industrial manufacturing process of safe and effective E40 under the Action, manufacturing work now focuses on further optimisation with respect to manufacturing strains, further commercial scale up and further downstream process optimisations, with early indications suggesting further yield improvements and economic efficiencies are achievable in the near-term.
Combining these achievements, the remaining task to commercialisation of a Phase 1a/1b trial has been significantly de-risked, having successfully demonstrated safety, efficacy and the ability to manufacture what is likely to be the first (and best) in class enzymatic therapy for the prevention of unintentional symptomatic gluten exposure in Celiac Disease.
