Synthetic alpha V beta 3 antagonists inhibit insulin-like growth factor-I-stimulated smooth muscle cell migration and replication

Porcine aortic smooth cells respond to insulin-like growth factor-I (IGF-I) with increases in DNA synthesis and cell migration. Because ligand occupan cy of the alpha V beta 3 integrin has been shown to be necessary for IGF-I to stimulate maximal increases in both processes, we determined whether syn thetic alpha V beta 3 antagonists could inhibit IGF-I-stimulated actions on this cell type. Low-molecular-weight compounds that had been selected base d on their ability to compete with vitronectin for binding to purified huma n alpha V beta 3 in vitro were analyzed for their ability to compete with I -125-kistrin (a known ligand for porcine alpha V beta 3) for binding to por cine alpha V beta 3. Nine compounds were screened, and five were found to b e potent competitive inhibitors. The most potent compound, SC-69000, result ed in 88% competition at 10(-7) M and was nearly equipotent with echistatin . The compounds that were the most potent inhibitors of kistrin binding wer e tested for their capacity to inhibit the cell migration response to IGF-I . Three compounds caused between 81-88% inhibition of IGF-I-stimulated migr ation at 10-7 M. To determine whether these compounds could inhibit other I GF-I-stimulated actions, their ability to inhibit IGF-I-stimulated [H-3]-th ymidine incorporation into DNA was analyzed. The four compounds that were t he most patent inhibitors of cell migration also inhibited IGF-I-stimulated DNA replication. IGF-I stimulates the synthesis of IGF binding protein-5 b y these cells. Preincubation with the four most active compounds also resul ted in significant inhibition of the ability of IGF-I to stimulate IGF bind ing protein-5 synthesis. alpha V beta 3 occupancy by the ligand vitronectin has been shown to enhance the capacity of IGF-I to activate its receptor t yrosine kinase. The four most active compounds were shown to inhibit IGF-I- stimulated IGF-I receptor autophosphorylation. These findings suggest that blockade of ligand occupancy of the alpha V beta 3 integrin globally inhibi ts several IGF-I-stimulated biologic actions and that synthetic inhibitors are very active in this regard. Because these compounds can be administered to whole animals, they should be very useful in determining whether blocki ng alpha V beta(3) occupancy in vivo results in alteration in esponsiveness to IGF-I.