The central hypothesis of this research project is that protein-protein interactions can be replicated or disrupted by stabilized peptides that have been identified via the identification of pharmacophores of small peptides that interact with CDK4/6. The basis for this hypothesis is formed by preliminary data generated by this lab that demonstrates small molecule inhibition of CDK4/6 can be achieved by applying peptide-based structure-function studies for pharmacophore identification. The rationale for the project is that stabilized peptides derived from these pharmacophores can be identified and evaluated on a faster basis than de novo synthesis of a series of compounds.
The project has three specific aims:
- Aim 1: Determine structure-function relationships of overlapping peptides derived from p16INK4a that inhibit the activity of CDK4/6 and identifying stabilized peptides that inhibit CDK4/6. The pharmacophore(s) of mutated peptides from p16INK4a will be assessed via time-resolved fluorescent resonance energy transfer assays and nuclear magnetic resonance spectroscopy to elucidate which residues are important to bind CDK4 and/or CDK6. Stabilized peptides will be evaluated for inhibitory activity toward CDK4/6.
- Aim 2: In vitro functional studies will be used to evaluate bioactivities of stabilized peptides. Various in vitro assays will be used to determine the functional efficacy of designed stabilized peptides.
- Aim 3: In vitro ADME studies and in vivo ADME and efficacy studies to evaluate the cell permeability, delivery, and efficacy of stabilized peptides. Stabilized peptides will be evaluated for “drug-like” qualities.