Author ORCID Identifier

0000-0002-7838-7116

Defense Date

2026

Document Type

Dissertation

Degree Name

Doctor of Philosophy

Department

Pharmaceutical Sciences

First Advisor

Yan Zhang

Abstract

Despite advances in cancer therapy, late-stage tumors frequently develop resistance and relapse, driven by cancer stem cells (CSCs). FOSL1 is overexpressed in CSCs and promotes invasion, survival, and chemoresistance; yet no FOSL1 inhibitor has entered the clinic. T-5224 suppresses HNSCC tumor growth and metastasis, but its limited potency restricts its utility. In the first project, we used a deconstruction-reconstruction-elaboration approach to design T-5224 11 analogs, identifying four that bind the FOSL1/JUN heterodimer. The benzoisoxazole is required for interaction, whereas the benzophenone is key for binding-site occupancy. Among the analogs, compound 3 downregulated FOSL1, inhibited HNSCC cell proliferation  ~tenfold more than T-5224, and reduced migration and CSC-sphere formation. It suppressed tumor growth at 50 mg/kg, comparable to T-5224, with no significant weight loss in xenograft mouse models.

PROTACs have been shown to eliminate CSCs and suppress HNSCC growth. In the second project, CRBN and VHL E3 ligase ligands with different PEG linker lengths were synthesized using T-5224. Binding studies showed that each E3 ligase required a distinct optimal linker length, necessitating empirical optimization. In the CRBN series, shorter linkers showed stronger binding; however, VZPT059, with a linker length (n=7), achieved the strongest degradation (DC50 ~0.4 µM; Dmax 99%) compared to VZPT052. VHL PROTACs with PEG5 or PEG6 linkers also degraded effectively. They inhibited proliferation, migration, and stem-cell sphere formation, showing 8–10-fold selectivity. In a mouse model, VZPT059 suppressed tumors and depleted active FOSL. Overall, targeting FOSL1 effectively addressed both the tumor and resistant stem cells that drive resistance and recurrence.

Rights

© Sarita Pandit

Is Part Of

VCU University Archives

Is Part Of

VCU Theses and Dissertations

Date of Submission

8-3-2026

Available for download on Saturday, August 02, 2031

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