BACKGROUND: We have recently released the structure of a class of quinolin-2-(1H)-on-3-carboxamide derivatives and among them; the drug A2 has the highest CB2 receptor affinity and selectivity. OBJECTIVE: In this work we assessed the immuno-modulatory properties of A2 in lymphocytes isolated from peripheral blood of multiple sclerosis patients and healthy donors. METHODS: Cell proliferative response was measured by 3H-thymidine incorporation, cell viability and apoptosis by trypan blue, annexin V staining and western blot. Cell activation was investigated by flow cytometry and molecular pathways by western blot. RESULTS: A2 exerted anti-proliferative effects with down-regulation of TNF- ?, IL-10 and Rantes in both cell types. No relevant changes were observed in cell viability between the two cell types. In cells form healthy subjects, A2 did not induce apoptosis, inhibited the cell cycle and similarly down-regulated in CD4+T the markers CD69, CD25, CD49d and CD54. Indeed, A2 also inhibited the phosphorylation of Akt, NF-kB, IKK?/?, ERK and blocked the expressions of Cox-2 and CB2 receptor. Differently, in cells from patients, A2 did not affect CD49d, while potently blocked CD54 expression. The inhibitory effects of A2 on Akt and Cox-2 expression were confirmed, whereas unchanged level of the CB2 receptor was observed in these cells. CONCLUSION: We reported similar effects of A2 in both cell types; however, a different mechanism of action might be suggested in cell from patients concerning cell activation and CB2 receptor expression. Overall, these data suggest an anti-inflammatory profile of A2 with potential implication in multiple sclerosis.
Immuno-modulatory Properties of a Quinolin-2-(1H)-on-3-carboxamide Derivative: Relevance in Multiple Sclerosis.
Laezza C;
2016
Abstract
BACKGROUND: We have recently released the structure of a class of quinolin-2-(1H)-on-3-carboxamide derivatives and among them; the drug A2 has the highest CB2 receptor affinity and selectivity. OBJECTIVE: In this work we assessed the immuno-modulatory properties of A2 in lymphocytes isolated from peripheral blood of multiple sclerosis patients and healthy donors. METHODS: Cell proliferative response was measured by 3H-thymidine incorporation, cell viability and apoptosis by trypan blue, annexin V staining and western blot. Cell activation was investigated by flow cytometry and molecular pathways by western blot. RESULTS: A2 exerted anti-proliferative effects with down-regulation of TNF- ?, IL-10 and Rantes in both cell types. No relevant changes were observed in cell viability between the two cell types. In cells form healthy subjects, A2 did not induce apoptosis, inhibited the cell cycle and similarly down-regulated in CD4+T the markers CD69, CD25, CD49d and CD54. Indeed, A2 also inhibited the phosphorylation of Akt, NF-kB, IKK?/?, ERK and blocked the expressions of Cox-2 and CB2 receptor. Differently, in cells from patients, A2 did not affect CD49d, while potently blocked CD54 expression. The inhibitory effects of A2 on Akt and Cox-2 expression were confirmed, whereas unchanged level of the CB2 receptor was observed in these cells. CONCLUSION: We reported similar effects of A2 in both cell types; however, a different mechanism of action might be suggested in cell from patients concerning cell activation and CB2 receptor expression. Overall, these data suggest an anti-inflammatory profile of A2 with potential implication in multiple sclerosis.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
