In vitro pharmacological characterization of standard and new lysophosphatidic acid receptor antagonists using dynamic mass redistribution assay

by Selleckchem | Jan 19 2024

Lysophosphatidic acid (LPA) is a bioactive phospholipid that acts as an agonist of six G protein-coupled receptors named LPA receptors (LPA1-6). LPA elicits diverse intracellular events and modulates several biological functions, including cell proliferation, migration, and invasion. Overactivation of the LPA-LPA receptor system is reported to be involved in several pathologies, including cancer, neuropathic pain, fibrotic diseases, atherosclerosis, and type 2 diabetes. Thus, LPA receptor modulators may be clinically relevant in numerous diseases, making the identification and pharmacodynamic characterization of new LPA receptor ligands of strong interest. In the present work, label-free dynamic mass redistribution (DMR) assay has been used to evaluate the pharmacological activity of some LPA1 and LPA2 standard antagonists at the recombinant human LPA1 and LPA2 receptors. These results are compared to those obtained in parallel experiments with the calcium mobilization assay. Additionally, the same experimental protocol has been used for the pharmacological characterization of the new compound CHI. KI 16425, RO 6842262, and BMS-986020 behaved as LPA1 inverse agonists in DMR experiments and as LPA1 antagonists in calcium mobilization assays. Amgen compound 35 behaved as an LPA2 antagonist, while Merck compound 20 from WO2012028243 was detected as an LPA2 inverse agonist using the DMR test. Of note, for all the compounds, similar potency values were estimated by DMR and calcium assay. The new compound CHI was found to be an LPA1 inverse agonist, but with potency lower than that of the standard compounds. In conclusion, we have demonstrated that DMR assay can be successfully used to characterize LPA1 and LPA2 ligands. Compared to the classical calcium mobilization assay, DMR offers some advantages, in particular allowing the identification of inverse agonists. Finally, in the frame of this study, a new LPA1 inverse agonist has been identified.

Comments:

This is fascinating! The versatility of Lysophosphatidic acid (LPA) and its receptors in modulating various biological functions certainly underscores their potential significance in numerous diseases. The utilization of label-free dynamic mass redistribution (DMR) assays in evaluating the pharmacological activity of LPA1 and LPA2 antagonists, along with the comparison to calcium mobilization assays, presents a promising approach.

It's intriguing that the DMR assay not only validated known antagonists but also revealed inverse agonists, offering a more comprehensive understanding of the compounds' actions. The ability to identify inverse agonists is particularly noteworthy as they can have distinct therapeutic implications compared to antagonists.

The discovery of a new LPA1 inverse agonist, CHI, albeit with lower potency, is a significant outcome of this study. This finding potentially broadens the scope for developing more targeted and effective therapeutic interventions.

The advantages of DMR over traditional methods, such as calcium mobilization assays, highlight its potential in drug discovery and characterization. The consistency in potency values estimated by both assays further validates the reliability of the DMR assay in this context.

Overall, this study not only sheds light on the pharmacological characterization of LPA1 and LPA2 ligands but also emphasizes the potential of DMR assays in advancing drug development for conditions linked to the LPA-LPA receptor system dysfunction.