Apolipoprotein E, a lipid carrier protein, exists as three isoforms: apoE2, apoE3, and apoE4. The apoE4 protein is less thermally stable than apoE2 and apoE3. Genome-wide association studies indicate that the possession of two E4 alleles is a strong genetic risk factor for late-onset Alzheimer’s disease. NMR-based screening on the N-terminal domain of apoE4 identified a fragment binder that was subsequently, using SBDD, elaborated into a single-digit micromolar stabilizer.

Post-translational functionalization of most secreted and transmembrane proteins requires co-translational translocation to the ER through Sec61. Translocation is negotiated by protein interactions between Sec61 and unique signal sequences specific to each translating protein. Disruption of these interactions in specific or multi-signal sequence fashion presents an opportunity to modulate protein homeostasis toward therapeutic benefit. Development of signal and multi-signal sequence selective Sec61 inhibitors as novel anti-cancer agents will be discussed.

BK and JC viruses reactivate during immunosuppression, resulting in nephropathy or multifocal leukoencephalopathy, respectively. To establish anti-viral MOAs involving viral protein-protein interactions, we examined capsid proteins VP1 and VP2 and identified a VP2-derived peptide with antiviral
activity. With biophysical and biochemical screens, we identified hits that bind VP1 and these compounds inhibit a VP1 and VP2 interaction in vitro.

Understanding how proteins interact is crucial to finding new drug candidates that can effectively disrupt or modify your PPI targets. Among the many biophysical techniques that have accelerated this process for scientists, surface plasmon resonance (SPR) is an industry standard that is both sensitive and powerful, enabling rapid and label-free detection of a wide range of molecular interactions. Yet, there has been minimal innovation in SPR platforms to date, considerably limiting their use and accessibility. Join us as we discuss how we’ve integrated digital microfluidics technology and AI-powered intelligence with SPR, and how this will eliminate challenges in efficiency and optimization to help you go-to-market faster.

A high-throughput MS platform was used to accurately detect and quantitate different covalent modifications of proteins, including KRAS G12C, which contain one or more reactive cysteines, lysines, or other nucleophilic residues. We employed the Agilent RapidFire system to rapidly quantitate the extent of covalent protein inhibitor adduct formation by MS for several proteins, including KRAS G12C and human serum albumin. We used this approach to screen large numbers of potential covalent inhibitors in an automated fashion and to test medicinal chemistry compounds as part of a regular lead optimization cycle for KRAS G12C.

MRTX849 is an irreversible, covalent inhibitor of KRASG12C currently undergoing clinical investigation in cancer patients with this mutation. This compound binds in the switch-II pocket of GDP-bound KRAS, locking the protein in the inactive state. Previously, we have described the structure-based design of the in vivo tool compound MRTX1257, and this talk will highlight the liabilities of this tool molecule and the strategies utilized for its final optimization to MRTX849.

The NCI RAS Initiative, led by scientific director, Frank McCormick of UCSF, combines novel drug discovery approaches (covalent tethering, computational modeling, and structure-guided fragment-based screening) to identify new compounds that directly target KRAS and its oncogenic mutants. These techniques provide a potential set of new targets in RAS drug discovery which have not yet been fully explored and will be discussed in this presentation.

The BCL-2 family protein BAX is a critical effector of apoptotic cell death in response to a diverse range of stimuli. My presentation will discuss the use of NMR and biochemical methods to screen and characterize the first inhibitors of inactive BAX that bind to a previously unrecognized allosteric pocket. Structure-based and mechanistic insights, cell-based and preclinical in vivo studies with this challenging PPI target will be discussed.

Disruption of intracellular calcium ion homeostasis leads to the unfolded-protein response and endoplasmic reticulum stress. These phenomena are recognized as causal features of major diseases such as diabetes and neurodegeneration. Herein we describe the use of FRET screening techniques for the discovery and optimization of small molecule calcium-handling modulators and their development to deliver drug candidates for these diseases.