Degradation of intrinsically disordered proteins using proteolysis targeting nanobody conjugate

Intrinsically disordered proteins (IDPs) play crucial roles in diverse cellular processes and are implicated in numerous diseases. However, due to the inherently unstructured nature, it is challenging to modulate their function using small molecule degraders or inhibitors. To address this, we introduced proteolysis targeting nanobody conjugate (PROTNC) composed of a nanobody that binds to an IDP, an E3 ligase ligand, and a cyclic cell-penetrating peptide for efficient intracellular delivery. We first showed a general-purpose PROTNC system capable to degrade different endogenous IDPs based on a co-condensate formation mechanism. We also introduced a straightforward strategy to degrade individual IDPs using specific PROTNCs equipped with a nanobody recognizing that particular IDP. Notably, we demonstrated the degradation of TPX2 which is an undruggable oncogenic IDP responsible for microtubule nucleation. This targeted degradation effectively inhibited cancer cell proliferation by disrupting bipolar mitotic spindle assembly in metaphase and directed cells to apoptotic pathway.