Bispecific nanobody® as new pharmacological drug for the selective
inhibition of Trypsin-3
- Melissa DAVID,
- Anais Faihy,
- Corinne Rolland,
- Anissa Edir,
- Astrid Canivet,
- Laetitia Ligat,
- mireille sebbag,
- Nathalie Vergnolle,
- Aurelien Olichon,
- Celine Deraison
Melissa DAVID
IRSD, Université de Toulouse, INSERM, INRAE, ENVT, Univ Toulouse III - Paul Sabatier (UPS), Toulouse, France
Author ProfileAnais Faihy
Université de Toulouse, Inserm, CNRS, Université Toulouse III-Paul Sabatier, Centre de Recherches en Cancérologie de Toulouse, Toulouse, France
Author ProfileAstrid Canivet
IRSD, Université de Toulouse, INSERM, INRAE, ENVT, Univ Toulouse III - Paul Sabatier (UPS), Toulouse, France
Author ProfileLaetitia Ligat
2Université de Toulouse, Inserm, CNRS, Université Toulouse III-Paul Sabatier, Centre de Recherches en Cancérologie de Toulouse, Toulouse, France
Author ProfileAurelien Olichon
Université de Toulouse, Inserm, CNRS, Université Toulouse III-Paul Sabatier, Centre de Recherches en Cancérologie de Toulouse, Toulouse, France
Author ProfileCeline Deraison
Inserm U1220
Corresponding Author:celine.deraison@inserm.fr
Author ProfileAbstract
Background and Purpose Proteolytic balance is dysregulated in many
diseases, with proteases playing critical roles in pathological
pathways. A high level of Trypsin-3 expression has been implicated as a
significant mediator of tumour progression and metastasis and this
protease is associated with poor prognosis for patient in various
cancers. Therefore, Trypsin-3 inhibition has emerged as a promising
therapeutic target. However, no physiological or pharmacological
inhibitor has yet been described that specifically target Trypsin-3. A
major challenge in developing druggable inhibitor for this protease lies
in achieving enough selectivity, as proteases belong to a large
enzymatic family with close homologues that share similarities in their
three-dimensional folding of their active conformation. Experimental
Approach An advanced screening strategy of a large library of synthetic
humanized nanobodies was employed to isolate highly selective
recombinant antibodies targeting the active conformation of Trypsin-3.
Among five hits, we combined two domains with distinct paratopes and
inhibitory mechanisms to generate a macrodrug candidate capable to
efficiently block the Trypsin-3 activity. Key Results This bispecific
nanobody demonstrated exceptionally high selectivity and affinity
towards Trypsin-3 in vitro, as well as a strong ability to inhibit
cancer cell migration ex vivo on PC-3 cancer cell line. Conclusion and
Implications. This study underscores the versatility and potential of
synthetic nanobody engineering in the development of very selective
protease inhibitors, paving the way for their consideration as drug
candidates for clinical development.01 Dec 2024Submitted to British Journal of Pharmacology 03 Dec 2024Submission Checks Completed
03 Dec 2024Assigned to Editor
03 Dec 2024Review(s) Completed, Editorial Evaluation Pending
07 Dec 2024Reviewer(s) Assigned