Kartogenin was shown to promote robust chondrocyte differentiation from primary human mesenchymal stem cells/MSCs [EC50 = 100 nM]. Kartogenin-induced chondrocytes from hMSCs express typical set of chondrogenic genes, form chondrocyte nodules, and exhibit characteristic chondrocyte functions. In an in vitro model mimicking cytokine-induced damage during osteoarthritis (OA), 1-5 µM of Kartogenin treatment inhibited nitric oxide (NO) and glycosaminoglycans (GAG) release. Direct Intra-articular (IA) administration of Kartogenin promoted cartilage repair in collagenase and surgery induced OA models in mice, and alleviated OA-induced joint pain. Mechanism of action studies showed that Kartogenin interacts with filamin A (FLNA) and results in the release of transcription factor CBFβ to the nucleus to regulate chondrogenic gene expression.