Rational Design and Synthesis of Novel Dual PROTACs for Simultaneous Degradation of EGFR and PARP

Abstract

Inspired by the success of dual targeting drugs, especially bispecific antibodies, we propose to combine the concept of protac and dual targeting to design and synthesize dual protac molecules with the function of degrading two completely different types of targets simultaneously. A library of novel dual targeting protac molecules have been rationally designed and prepared. A convergent synthetic strategy has been utilized to achieve high synthetic efficiency. These dual protac structures are characterized by using trifunctional natural amino acids as star-type core linkers to connect two independent inhibitors and E3 ligands together. In this study, gefitinib, olaparib, and CRBN or VHL E3 ligand were used as substrates to synthesize novel dual protacs. They successfully degraded both EGFR and PARP simultaneously in cancer cells. Being the first successful example of dual protacs, this technique will greatly widen the range of application of the protac method and open up a new field for drug discovery. File list (2) download file view on ChemRxiv dual_protac manuscript.pdf (1.02 MiB) download file view on ChemRxiv Supplementary Information.pdf (4.08 MiB)

Trivalent PROTACs enhance protein degradation via combined avidity and cooperativity

“…One might imagine two subunits within a multi-protein complex, or even two distinct proteins (albeit at the expense of avidity), could be recruited either simultaneously or independently to the E3 ligase. While this manuscript was in advanced stages of revision, a dual-target PROTAC compound consisting of three warheads was reported, 48 however not shown to engage all three warheads simultaneously as we report here.…”

Targeted protein degradation: mechanisms, strategies and application

“… 42 PROTAC and molecular glue are two major technologies that rely on the UPS for the degradation of protein of interest (POI), and will be the focus of our discussion (Table 1 ). Additionally, many PROTAC-based technologies, including selective androgen receptor degrader (SARD) 43 , 44 , Hydrophobic tagging (HyT), 45 – 47 TF-PROTAC, 48 dual-PROTAC, 49 and selective estrogen receptor degrader (SERD), 50 – 54 have recently emerged (Table 1 ).…”

Strategies for designing proteolysis targeting chimaeras (PROTACs)

“…86 They designed a tetrazine-tagged thalidomide derivative (46) and trans-cyclo-octene-tagged JQ1 derivative (47), which are cell penetrable (Figure 7A). These derivatives can react rapidly to form PROTACs (48) within cells in which they had not previously shown degradation activity due to low penetration into cells. When cells were treated with these two fragments consecutively, the expression level of BRD4 was dramatically downregulated in a time-and concentrationdependent manner, indicating that these two fragments can react with each other to form PROTACs in cells.…”

“…Therefore, it was removed (compound 9), and the phenolic hydroxyl group was suitable for the introduction of linkers (Figure 3). 48 Irreversible POI ligands can be modified to design either reversible PROTACs or irreversible PROTACs. Theoretically, irreversible inhibitors exhibit stronger affinity and higher selectivity than reversible inhibitors because irreversible inhibitors can form covalent bonds with specific amino acids in a time-dependent manner.…”

“…To validate this hypothesis, Li's group designed dual-targeting PROTACs that simultaneously degraded EGFR and PARP. 48 The EGFR inhibitor gefitinib, PARP inhibitor olaparib, and E3 ligase ligand were merged by star-type linkers (Figure 21), which featured three potential reactive sites (an esterification site, an amide condensation site, and a click reaction site) based on the biocompatible natural amino acids tyrosine tool as an external control factor. In recent years, light has been widely applied to regulate biological pathways with high spatiotemporal resolution.…”