生物活性

• 描述Vps34, a PI3K class III isoform, plays an important role in autophagy. SAR405 is the first potent and specific Vps34 inhibitor with KD value of 1.5 nM. Inhibition of Vps34 kinase activity by SAR405 affects both late endosome-lysosome compartments and prevents autophagy. Treatment with 1μM SAR405 triggered the relocalization of GFP-FYVE inside the HeLa cells stably transfected with GFP-FYVE, indicating the inhibition of PtdIns3P (a substrate specifically catalyzed by Vps34) formation by SAR405. A induction of late endosome-lysosome swelling could be observed in RKO cells treated with 10 μM SAR405 for 16 h. Dose-dependent accumulation of p62 could triggered by SAR405 at concentration ranging in 0.1-10μM for 24h in RKO cells. These suggested that SAR405 could affect vesicle trafficking in cells. Treatment with SAR405 at concentration ranging in 0.1-10μM combined with 10 μM hydroxychloroquine could concentration-dependently prevent starvation-induced autophagy in both HeLa and H1299 cells, shown by decreased level of LC3-II compared with hydroxychloroquine-treated group alone. This prevention by SAR405 could also observed in autophagy induced by mTOR inhibitor AZD8055 and synergizes with everolimus to H1299 cell growth inhibition. Ex vivo treatment of WT mouse or human platelets with 1μM SAR405 induced abnormal secretion and affected thrombus growth at arterial shear rate. SAR405 infusion into basolateral amygdala (1μM SAR405 was infused into the BLA at a rate of 0.5 μL/min with a total volume of 0.5 μL/side) could impaire long-term memory through autophagy inhibition. The trafficking of GABAARs following fear conditioning was disrupted by SAR405. The decreased frequency and amplitude of mIPSCs induced by fear conditioning were also reversed by SAR405. SAR405 could suppress the upregulation of the interaction of GABAA receptor associated protein with GABAAR γ2 subunit after fear conditioning. These suggested SAR405 could disrupt auditory fear memory consolidation of mice via facilitation of inhibitory neurotransmission in basolateral amygdala.
• 作用机制SAR405 has an exquisite protein and lipid kinase selectivity profile due to its unique binding mode and molecular interactions within the ATP binding cleft of human Vps34.^[1]