Periodic Reporting for period 1 - MirrorBody (Discovery, total chemical synthesis, and validation of “mirror-image” D-Nanobodies)
Periodo di rendicontazione: 2022-08-01 al 2024-07-31
Therapeutic proteins are the most rapidly growing class of drugs, and antibody-based biologics represent their vast majority. Antibodies are excellent tools for target binding and for disrupting protein-protein interactions in terms of potency and specificity, but they have limitations such as poor tissue and cellular penetration and complex manufacturing. Small nature-derived and engineered proteins can retain antibody binding affinity and specificity; and thanks to their reduced size can overcome penetration issues and can be accessed by chemical synthesis, making them extremely versatile to introduce tailored modifications, but a key limitation that is linked to all therapeutic proteins is their immunogenicity. Immunogenicity of therapeutic proteins involves their tendency to trigger an unwanted immune response against themselves, for example by producing antibodies that bind to the proteins and reduce or eliminate their therapeutic effects, but also leading to potentially life-threatening complications. In this scenario D-proteins, completely composed of D-amino acids, have the unique advantage of having an extremely low immunogenic potential. Although these properties have been known for almost 60 years, currently there are very few examples of antibody-like D-proteins in the literature, the main bottleneck being the lack of efficient discovery processes. The overall objective of this project is to explore the potential of a novel, multidisciplinary approach for the discovery of D-proteins as for molecular targeting that could lead to a novel generation of biomedical tools.
The activities performed throughout this project revolve around the proposed new approach for D-protein discovery, which has not yet been published. In pursuing this ultimate and ambitious goal, the disclosable achievements involve synthetic methodologies for protein chemical synthesis, including, for example, the development of a new solubility tag to overcome peptide insolubility and aggregation issues (unpublished results), which is key for this project. Another achievement obtained during this project that concerns a synthetic methodology aspect involves the development of a dual, site-selective functionalization approach to generate bimodal antibody-like small proteins.
This project has yielded compelling results that have significant implications for both academia and industry. This research has the potential to provide a major advancement in the field of D-protein therapeutics, which have the potential to overcome key limitations that are associated with their immunogenic potential. This work paves the way for developing a new discovery approach to develop D-proteins as tools for biomedicine. However, our journey towards the ultimate goal has been marked by challenges. In particular, we encountered difficulties for the chemical synthesis of proteins, which posed significant obstacles to our progress. We undertook the development of innovative tools and methodologies to overcome these challenges. Through iterative experimentation and problem-solving, we successfully devised novel approaches that enabled us to circumvent some limitations of traditional protein synthesis approaches. These developments not only represent significant achievements in their own right but also underscore the resilience and adaptability of our research team in the face of adversity. Moving forward, these tools will serve as invaluable assets in our pursuit of further advancements.