Affinity Measurement of Non-covalent Interactions of the Covalent KRAS G12C GDP Inhibitor MRTX849 to RAS Isoforms...

Area of Science:

• Biochemistry
• Molecular Biology
• Biophysics

Background:

• Surface Plasmon Resonance (SPR) is a label-free optical technique for real-time biomolecular interaction analysis.
• SPR is versatile, applicable to proteins, nucleic acids, and small molecules, measuring affinity, association, and dissociation rates.
• RAS proteins (HRAS, KRAS, NRAS) are crucial in cell signaling and cancer, making them important drug targets.

Purpose of the Study:

• To describe a general method for measuring the binding affinity of small molecule drugs to RAS proteins using SPR.
• To quantify the real-time interaction kinetics between MRTX849 and GDP-bound HRAS, KRAS, and NRAS.

Main Methods:

• Utilized Surface Plasmon Resonance (SPR) spectroscopy for label-free, real-time kinetic analysis.
• Immobilized GDP-bound HRAS, KRAS, and NRAS onto SPR sensor chips.
• Flowed the small molecule drug MRTX849 over the immobilized proteins to measure binding.

Main Results:

• Successfully measured the affinity of MRTX849 to GDP-bound HRAS, KRAS, and NRAS.
• Quantified association (kon) and dissociation (koff) rates for these interactions.
• Demonstrated the utility of SPR for characterizing small molecule drug interactions with RAS isoforms.

Conclusions:

• SPR provides a robust and versatile platform for characterizing small molecule drug interactions with RAS proteins.
• The described method enables real-time affinity and kinetic measurements crucial for drug discovery and development.
• Understanding MRTX849's binding kinetics to HRAS, KRAS, and NRAS is vital for its therapeutic potential in targeting these oncogenic proteins.