Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full length human Beclin-1 (hBeclin-1-Myc/DDK; +) and/or mouse ULK1 protein (mULK1; +). Transfected cells were treated with SBI-0206965 (2 hrs, indicated concentrations). Western blot was performed using Phospho-Beclin-1 (Ser15) (D4B7R) Rabbit mAb #84966 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).Learn more about how we get our images
Chemical structure of SBI-0206965Learn more about how we get our images
SBI-0206965 is supplied as a lyophilized powder. For a 10 mM stock, reconstitute 5 mg of powder in 1.02 ml of DMSO. Working concentrations and length of treatment can vary depending on the desired effect, but it is typically at 10-50 μM for 1-72 hours.
Store lyophilized or in solution at -20ºC, desiccated. In lyophilized form, the chemical is stable for 24 months. Once in solution, use within 3 months to prevent loss of potency. Aliquot to avoid multiple freeze/thaw cycles.
SBI-0206965 is a potent, selective and cell permeable inhibitor of the autophagy-promoting serine/threonine kinase ULK1 (1). ULK1 is an essential protein for induction of autophagy by nutrient starvation and mTOR inhibition (2). ULK1 is recripocally regulated by multple phosphorylation sites via AMPK and mTOR, such that ULK1 is activated by AMPK and inhibited by mTOR (3, 4). By in vitro kinase assays, SBI-0206965 inhibits ULK1 with an IC50 of 108 nM and to a lesser extent ULK2 with an IC50 of 711 nM (1). In a panel of 456 kinases, when tested at 10 μM, SBI-0206965 showed remarkable selectivity and only inhibited 10 kinases, including ULK1 and ULK2. Other kinase that were inhibited included FAK, FLT3, Src, and Jak3. Only FLT3 and FAK had an IC50 similar to ULK1 when tested in in vitro kinase assays. SBI-020695 inhibits survival following nutrient deprivation. Furthermore, SBI-0206965 inhibits autophagy induced by mTOR inhibition and converts the cytostatic response to mTOR inhibition to a cytotoxic apoptotic response.
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