|
|||||
|
|
Combining Solid-State NMR with Structural and Biophysical Techniques to Design Challenging Protein-Drug Conjugates |
|
| Authors: | Dr. Linda Cerofolini Kristian Vasa Elisa Bianconi Maria Salobehaj Giulia Cappelli Alice Bonciani Giulia Licciardi Dr. Anna Pérez-Ràfols Luis Padilla-Cortés Sabrina Antonacci Dr. Domenico Rizzo Prof. Dr. Enrico Ravera Prof. Dr. Caterina Viglianisi Prof. Dr. Vito Calderone Prof. Dr. Giacomo Parigi Prof. Claudio Luchinat Prof. Dr. Antonio Macchiarulo Prof. Stefano Menichetti Prof. Dr. Marco Fragai |
| Affiliation: | 1. Magnetic Resonance Centre (CERM), University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, Italy Consorzio Interuniversitario Risonanze Magnetiche di Metalloproteine (CIRMMP), Via L. Sacconi 6, 50019 Sesto Fiorentino, Italy Department of Chemistry “Ugo Schiff”, University of Florence, Via della Lastruccia 3-13, 50019 Sesto Fiorentino, Italy These authors contributed equally to this work.;2. Department of Chemistry “Ugo Schiff”, University of Florence, Via della Lastruccia 3-13, 50019 Sesto Fiorentino, Italy These authors contributed equally to this work.;3. Department of Pharmaceutical Sciences, University of Perugia, Via Fabretti n.48, 06123 Perugia, Italy These authors contributed equally to this work.;4. Magnetic Resonance Centre (CERM), University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, Italy Consorzio Interuniversitario Risonanze Magnetiche di Metalloproteine (CIRMMP), Via L. Sacconi 6, 50019 Sesto Fiorentino, Italy Department of Chemistry “Ugo Schiff”, University of Florence, Via della Lastruccia 3-13, 50019 Sesto Fiorentino, Italy;5. Department of Chemistry “Ugo Schiff”, University of Florence, Via della Lastruccia 3-13, 50019 Sesto Fiorentino, Italy;6. Magnetic Resonance Centre (CERM), University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, Italy Giotto Biotech s.r.l, Sesto Fiorentino, Via della Madonna del Piano 6, 50019 Florence, Italy;7. Magnetic Resonance Centre (CERM), University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, Italy Department of Chemistry “Ugo Schiff”, University of Florence, Via della Lastruccia 3-13, 50019 Sesto Fiorentino, Italy;8. Magnetic Resonance Centre (CERM), University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, Italy Consorzio Interuniversitario Risonanze Magnetiche di Metalloproteine (CIRMMP), Via L. Sacconi 6, 50019 Sesto Fiorentino, Italy Department of Chemistry “Ugo Schiff”, University of Florence, Via della Lastruccia 3-13, 50019 Sesto Fiorentino, Italy Giotto Biotech s.r.l, Sesto Fiorentino, Via della Madonna del Piano 6, 50019 Florence, Italy;9. Department of Pharmaceutical Sciences, University of Perugia, Via Fabretti n.48, 06123 Perugia, Italy;10. Magnetic Resonance Centre (CERM), University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, Italy |
| Abstract: | Several protein-drug conjugates are currently being used in cancer therapy. These conjugates rely on cytotoxic organic compounds that are covalently attached to the carrier proteins or that interact with them via non-covalent interactions. Human transthyretin (TTR), a physiological protein, has already been identified as a possible carrier protein for the delivery of cytotoxic drugs. Here we show the structure-guided development of a new stable cytotoxic molecule based on a known strong binder of TTR and a well-established anticancer drug. This example is used to demonstrate the importance of the integration of multiple biophysical and structural techniques, encompassing microscale thermophoresis, X-ray crystallography and NMR. In particular, we show that solid-state NMR has the ability to reveal effects caused by ligand binding which are more easily relatable to structural and dynamical alterations that impact the stability of macromolecular complexes. |
| Keywords: | Drug Delivery Drug Design NMR Spectroscopy Protein-Drug Conjugates Structural Biology |