Pharmacological characterisation of a tool αvβ1 integrin small molecule RGD-mimetic inhibitor

Compound 8 is a selective αvβ1 small molecule inhibitor that has been used in pre-clinical studies to identify and characterise the αvβ1 integrin as a potential target in fibrotic disease. In this study we further investigated the selectivity and pharmacokinetics of compound 8 to determine a link between the levels of αvβ1 engagement required to achieve in vivo pharmacodynamic efficacy. The selectivity of compound 8 for the arginyl-glycinyl-aspartic acid and β1 integrins was measured using purified integrin protein preparations in radioligand binding studies with both labelled ([³H]compound 8) and unlabelled versions. The pharmacokinetic profile of compound 8 was completed in in vitro blood protein binding assays and in in vivo studies using male C57BL/6 mouse following i.v. dosing. The high selectivity of compound 8 for αvβ1 over the other αv integrins was confirmed, however a reduced selectivity was demonstrated for the β1 integrin family, with high affinity observed for α4β1 (comparable to αvβ1), moderate affinity for α2β1, α3β1 and α8β1, and low affinity for α5β1 and α9β1. Compound 8 was shown to be cleared quickly from the blood with a short half-life of 0.5 h. In conclusion, the data in this study suggest that compound 8 has the potential to engage a number of integrins in vivo beyond αvβ1, that raises a degree of uncertainty regarding its mechanism of action in models of fibrotic disease.