Wortmannin, Wm (1R,3R,5S,9R,18S)-18-(methoxymethyl)-1,5-dimethyl-6,11,16-trioxo-13,17-dioxapentacyclo[10.6.1.0^{2,10}.0^{5,9}.0^{15,19}]nonadeca-2(10),12(19),14-trien-3-yl acetate CHEMBL428496 cid_312145 BDBM15234 NSC221019 Wortmannin
Ring-opened wortmannin analogue, 4 PX-866 (2R,3S,6E,15S,17R)-6-{[bis(prop-2-en-1-yl)amino]methylidene}-8-hydroxy-3-(methoxymethyl)-2,15-dimethyl-5,9,14-trioxo-4-oxatetracyclo[8.7.0.0^{2,7}.0^{11,15}]heptadeca-1(10),7-dien-17-yl acetate BDBM21035
- Yuan, H; Barnes, KR; Weissleder, R; Cantley, L; Josephson, L Covalent reactions of wortmannin under physiological conditions. Chem Biol 14: 321-8 (2007)
- Liu, Y; Jiang, N; Wu, J; Dai, W; Rosenblum, JS Polo-like kinases inhibited by wortmannin. Labeling site and downstream effects. J Biol Chem 282: 2505-11 (2007)
- Walker, EH; Pacold, ME; Perisic, O; Stephens, L; Hawkins, PT; Wymann, MP; Williams, RL Structural determinants of phosphoinositide 3-kinase inhibition by wortmannin, LY294002, quercetin, myricetin, and staurosporine. Mol Cell 6: 909-19 (2000)
- Giner, JL; Kehbein, KA; Cook, JA; Smith, MC; Vlahos, CJ; Badwey, JA Synthesis of fluorescent derivatives of wortmannin and demethoxyviridin as probes for phosphatidylinositol 3-kinase. Bioorg Med Chem Lett 16: 2518-21 (2006)
- In Vitro Enzymatic Activity Assay 1. Preparation of 1-Fold Kinase Buffer1) 1-fold kinase buffer40 mM Tris, pH 7.50.0055% Brij-3520 mM MgCl2 0.05 mM DTT2. Compound Preparation1) The detection starting concentration of the compound was 1 μM, and it was prepared to a 100-fold concentration, namely 100 μM. 2 μl of 10 compound was taken, 198 μl of 100% DMSO was added, and it was prepared into 100 μM compound solution. 100 μl of the compound of 100-fold concentration was added to a second well on a 96-well plate, and 60 μl of 100% DMSO was added to other wells. 30 μl of the compound was taken from the second well and added to a third well, it was diluted downwards sequentially by 3 times, and there was a total of 10 concentrations diluted.2) 100 μl of 100% DMSO and positive control wortmannin with the highest concentration (400 nM) were transferred to two empty wells as a Max well and a Min well respectively.3) 50 nl of the compound was transferred to a 384-well plate by Echo.3. Preparation of 2× Kinase Solution1) The 1-fold kinase buffer was used to prepare 2-fold DNA-PK kinase solution.2) 2.5 μl of the 2-fold kinase solution was transferred to a reaction well of the 384-well plate.3) It was shaken, mixed uniformly, and still placed at a room temperature.4. Preparation of 2× Substrate Solution1) The 1-fold kinase buffer was used to prepare 2-fold substrate solution.2) 2.5 μl of the 2-fold substrate solution was transferred to the reaction well of the 384-well plate for a starting reaction.3) It was shaken, and mixed uniformly.5. Kinase Reaction and Termination1) The 384-well plate was covered by a cover, and incubated at 28° C for 3 hours.2) 5 μl of ADP-Glo reagent was transferred, and incubated at 28° C. for 2 hours.6. Detection of Reaction Result1) 10 μl of a kinase detection reagent was transferred into the reaction well of the 384-well plate for terminating the reaction.2) It was still placed at the room temperature for 30 minutes.7. Data ReadingSample values were read on Envision.8. Inhibition Rate Calculation1) Data was copied from Envision.2) It was converted into inhibition rate data.Inhibition percentage=(max-conversion)/(max-min)*100. Wherein, max refers to the conversion rate of a DMSO control, min refers to the conversion rate of a control without enzyme activity, and conversion refers to the conversion rate of the test compound at each concentration.