Live-Cell Assays Reveal Two Reported Anti-Inflammation Molecules Fail to Engage Target Protein

Madison, WI USA. (October 25, 2023)

A new study uses key Promega technologies to evaluate reported anti-inflammation molecules in the context of live cells. Promega research scientists used three unique assays to evaluate seven inhibitors targeting the NLRP3 inflammasome, a hot target in drug development for its role in multiple inflammatory diseases. The team was able to rank the potency of five reported NLRP3 inhibitors and demonstrate that two failed to produce the intended biological effects. The paper is available open access from Cell Chemical Biology.

NLRP3 triggers critical inflammatory immune responses, but its dysregulation can result in autoinflammatory disease. This protein has attracted attention as a potential drug target for many diseases involving inflammation, and several candidates are progressing through pre-clinical studies and clinical trials

Promega research scientists used three unique technologies to analyze how reported NLRP3 inhibitors behaved in live human cells. The NanoBRET® Target Engagement Assay allowed the team to study inhibitor binding to NLRP3 in the natural cellular context. The Caspase-Glo® 1 Inflammasome Assay and Lumit™ IL-1β Immunoassay indicated whether the inhibitor produced the desired effects on inflammasome activity and inflammatory cytokine release. Based on the results of these assays, the team determined that two reported inhibitors did not engage NLRP3 or functionally inhibit the target pathway.

Promega Corporation is a leader in providing innovative solutions and technical support to the life sciences industry. The company’s portfolio of over 4,000 products supports a range of life science work across areas such as cell biology; DNA, RNA and protein analysis; drug development; human identification and molecular diagnostics. These tools and technologies have grown in their application over the last 45 years and are used today by scientists and technicians in labs for academic and government research, forensics, pharmaceuticals, clinical diagnostics and agricultural and environmental testing. Promega is headquartered in Madison, WI, USA with branches in 16 countries and over 50 global distributors.