OBJECTIVE I:
We established gut stability assays and bioassays to test activity at OTR. Using cutting-edge medicinal chemistry, we produced leads equipotent to OT, yet with a gut stability half-life of >24 h compared to 8 min of OT. Our leads are potent analgesics when given orally in IBS mouse models with a gut specific mode of action. These first-in-class oral and non-opioid-based leads were patented and form the basis of discussions with biotech.
MAIN OUTCOMES:
- gut stability assays & bioassays
- gut-stable bioactive drug leads
- new knowledge about OTR in gut disorders
- high-impact publications and patent
- ERC PoC grant on advancing drug leads
OBJECTIVE II:
The trefoil factor family (TFF) are peptides (TFF1-3) expressed throughout the gut with roles in gut protection and repair. To investigate the molecular mechanisms, we first needed a reliable way of producing them. Using a combination of solid-phase peptide synthesis and native chemical ligation we, for the first time, synthesised TFF1, TFF3 and their corresponding homodimers. We demonstrated that they were folded correctly and that TFF1 interacts with mucins. We revealed that TFF1&3 get digested in the gut but yield a bioactive metabolite. We identified a receptor for TFF1 and TFF3. Production of TFF2 was more challenging, and we had to pursue recombinant expression, semi-synthetic methods, and total chemical synthesis. We have now also control over TFF2 production. Isotope labelling of TFF2 to determine its 3D structure, resulted in a new technology of producing isotope-labelled amino acids.
MAIN OUTCOMES:
- Reliable production of TFF1-3 and TFF1/3 homodimers
- Discovery of a gut-stable bioactive metabolite of TFF1/3
- Discovery of target receptor of TFF1/3
- New mechanistic insights
- Key reviews and high impact publications
- ERC PoC grant to develop isotope-labelled amino acid production technology
OBJECTIVE III:
Venoms are a rich source of bioactive peptides and a main source in this project to identify new ligands that could accelerate wound healing or be used as gut-stable scaffolds for drug development.
We obtained venoms from venomous animals, fractionated them, and characterised the fractions by HPLC and mass spectrometry. We developed a medium-throughput pipeline to discover novel peptide ligands that are stable to gastric and intestinal digestion. Identified leads were synthesised and tested for gut wound healing and mode-of-action. While no direct wound healing activity was discovered, some of these ligands might serve as gut-stable scaffolds for drug development.
MAIN OUTCOMES:
- Establishment of cell lab and epithelial repair assays
- Identified gut-stable ligands using our new discovery platform
- PeptideMiner, a new tool for in-silico screening of peptide homologues
- Reviews and publications in venom peptide drug discovery
Our research outcomes resulted in invitations at international conferences, new collaborations, prizes and awards, and several articles covering our research.
