https://orcid.org/0000-0001-6887-0836
Figures & data
Figure 1. Function of wild-type IDH in homeostasis and activity of mutant IDH in disease. α-KG, α-ketoglutarate; 2-HG, D-2-hydroxyglutarate; D2HGDH, D-2-hydroxyglutarate dehydrogenase; IDH, isocitrate dehydrogenase; IDH2m, mutant IDH2.
Figure 2. Chemical structures of representative mutant IDH2R140Q inhibitors.
Figure 3. Rational design of novel target compound in series (I), 6a–l, series (II), 7a–l, and (III), 8a–d, taking enasidenib as a lead compound.
Scheme 1. Synthesis of target compounds in series (I), 6a–l and series (II), 7a–l. Reagents and conditions: i: Acetone, Na2CO3, 0–5 °C, 4 h; ii: Acetonitrile, hydrazine hydrate 99%, heat under reflux; iii: Ethanol, glacial acetic acid, and heat under reflux.
Scheme 2. Synthesis of compounds in series (III), 8a–d. Reagents and conditions: i: 4-Nitroacetophenone, ethanol, glacial acetic acid, reflux 8 h. ii: 5-Chloroisatin, ethanol, glacial acetic acid, reflux 8 h. iii: Acetonitrile, L-phenylalanine methyl ester hydrochloride, Na2CO3 solution, reflux 96 h. iv: Benzohydrazide, acetonitrile, and reflux 6 h.
Figure 4. 1H-1H Homonuclear 2 D NOESY spectrum for E-isomer of compound 6i.
Figure 5. Mean growth inhibition percentage (GI %) on 60 cancer cell lines for series (I) and series (II) in the single dose experiment.
Table 1. Preliminary anticancer effects of single dose (10 μM) of s-triazine derivatives 6a, 6c, 6d, 7a, 7 g, 7i, 7l, and 8b against 60 human subpanel cancer cell lines declared as the percentage cell growth inhibition (GI%).
Figure 6. Summary of structure–activity relationship (SAR) of series (I), (II), and (III) as anticancer agents against 60 human subpanel cancer cell lines relying on the values of mean GI%.
Table 2. Cytotoxic effects of five doses (0.01–100 μM) for compounds, 6a, 6c, 6d, 7g, 7l, and enasidenib towards 60 human subpanel cancer cell lines declared as GI50 (μM).
Table 3. In vitro inhibition of mutant and wild type IDH2 enzymes by selected target compounds.
Table 4. Cytotoxic activity of compound 6c and staurosporine against the normal kidney cells of human embryo (HEK-293).
Table 5. Cell cycle analysis of HL-60(TB) cells treated with compound 6c and DMSO as a negative control.
Figure 7. Effect of DMSO (upper two panels) and compound 6c (lower two panels) on the cell cycle distribution of HL-60(TB) cancer cell line.
Figure 8. Apoptosis assay on HL-60(TB) cancer cell line induced by DMSO (left panel) and compound 6c (right panel) the four quadrants identified as: LL: viable; LR: early apoptotic; UR: late apoptotic; UL: necrotic.
Figure 9. Summary of the Annexin V-FITC Apoptosis assay results of compound 6c and DMSO on the percentage of HL-60(TB) cells stained positive for Annexin V-FITC.
Figure 10. Impact of compound 6c on expression of active Caspase 3 and Caspase 9 levels in HL-60(TB) cancer cells.
Figure 11. Binding interaction of enasidenib (A) and compounds 6c (B), 6e (C), and 7c (D) inside IDH2R140Q allosteric site (PDB ID: 5I96). 2D pose binding of the compound (left), green lines (H-bond), pink lines (hydrophobic interactions), cyan lines (halogen bond), and 3D surface representation of the compound in the allosteric site (right).
Table 6. Docking results and interacting residues for inhibitors, 6c, 6e, 7c, and enasidenib in IDH2R140Q allosteric site (PDB ID: 5I96).
Figure 12. Alignment of compounds 6c (dark red), 6e (blue), and 7c (orange) and enasidenib (green) in the IDH2R140Q allosteric site viewing parallel layout.
Figure 13. Predicted BOILED-Egg for compounds 6a, 6c, 6d, 7g, and 7l. BBB: blood–brain barrier; HIА; human intestinal absorption; PGP+: P-glycoprotein substrate; PGP-: not P-glycoprotein substrate.
Table 7. Drug-likeness profiles for compounds 6a, 6c, 6d, 7g, and 7l and number of rules they fulfilled.
Figure 14. Radar charts for prediction of oral bioavailability profiles of compounds 6a, 6c, 6d, 7g, and 7l represented by red line, and the range of optimal property values are shown in pink.
