1383716-33-3

Vps34-IN-1 is a potent and selective inhibitor of phosphoinositide 3-kinase (PI3K) vacuolar protein sorting 34 (VPS34) (1). VPS34 inhibitors can be used to investigate autophagic processes (2).

vps34-in1 is a vps34 inhibitor.the vacuolar protein sorting 34 (vps34) class iii phosphoinositide 3-kinase (pi3k) phosphorylates phosphatidylinositol (ptdins) to generate ptdins(3)p that regulates membrane trafficking processes through its ability to recruit a subset of proteins possessing ptdins(3)p-binding phox homology (px) and fyve domains.

Steps for the synthesis of 1-((2-((2-chloropyridin-4-yl)amino)-4'-(cyclopropylmethyl)-[4,5'-bipyrimidinyl]-2'-yl)amino)-2-methylpropan-2-ol (16a): 4-amino-2-chloropyridine (53.3 mg, 0.415 mmol) was dissolved in THF (277 μL) under nitrogen protection and cooled to -78 °C. . LiHMDS (415 μL, 0.415 mmol) was slowly added dropwise to this solution. The reaction mixture was gradually warmed to room temperature and stirred continuously for 30 min. Subsequently, the reaction mixture was cooled to -78 °C again and 1-(4'-(cyclopropylmethyl)-2-(methylsulfinyl)-4,5'-bipyrimidin-2'-ylamino)-2-methylpropan-2-ol (50 mg, 0.138 mmol) was added. The reaction mixture was warmed up to room temperature and stirring was continued for 1 hour. The completion of the reaction was confirmed by LC/MS monitoring. The crude product was purified by reversed-phase HPLC [gradient elution: 30-100% organic phase, 15 min] followed by Biotage silica gel column chromatography [10 g SNAP column, elution gradient: 100% DCM to 12% MeOH/DCM] to afford the target product (29 mg, 49% yield).

vps34-in1 was identified to be able to inhibit in-vitro vps34, but did not significantly inhibit the activity of tested 340 protein kinases or 25 lipid kinases that include all isoforms of class i and class ii pi3ks. treatment of vps34-in1 to cells dose-dependently induced a quick dispersal of a specific ptdins(3)p-binding probe, without altering the ability of class i pi3k to regulate akt. moreover, vps34-in1 could also induce a quick ~50-60% loss of sgk3 phosphorylation within 1 min. however, vps34-in1 could not inhibit activity of the sgk2 isoform without a ptdins(3)p-binding px domain. in addition, the combination of vps34-in1 and class i pi3k inhibitor of gdc-0941 resulted in the reduced sgk3 activity ~80-90%. therefore, these data demonstrated that vps34-in1 would be a useful probe to delineate physiological roles of the vps34 and the combination of class i such as gdc-0941 and class iii such as vps34-in1 pi3k inhibitors could be used to better analyse the roles and regulation of the elusive class ii pi3k [1].

25 nm

[1] bago r et al. characterization of vps34-in1, a selective inhibitor of vps34, reveals that the phosphatidylinositol 3-phosphate-binding sgk3 protein kinase is a downstream target of class iii phosphoinositide 3-kinase. biochem j.2014 nov 1;463(3):413-27.