Discovery of dual tubulin-NEDDylation inhibitors with antiproliferative activity

Figures & data

Figure 1. Tubulin polymerisation inhibitors as anticancer agents.

Figure 2. Representative NEDDylation modulators.

Figure 3. Rational design of tubulin-NEDDylation inhibitors.

Scheme 1. Reagents and conditions: (i) 3,4,5-Trimethoxyaniline, CH₂Cl₂, triethylamine, rt, 12 h; (ii) Acyl chlorides, CH₂Cl₂, rt, 2 h; (iii) CS₂, Secondary amines, KOH, acetone, reflux, 7 h.

Table 1. Antiproliferative activity of compounds A1 and C1–C15.

Compound	HepG2	PC3	MCF7
A1	> 30	> 30	> 30
C1	2.07 ± 0.14	2.96 ± 0.75	4.63 ± 0.91
C2	2.34 ± 0.12	3.37 ± 1.26	3.57 ± 0.63
C3	3.50 ± 0.37	3.42 ± 0.50	5.67 ± 0.24
C4	10.16 ± 1.27	15.37 ± 1.25	14.31 ± 1.27
C5	7.12 ± 0.10	15.94 ± 0.43	10.52 ± 0.47
C6	5.66 ± 0.15	14.13 ± 1.90	12.56 ± 0.52
C7	12.68 ± 0.81	17.62 ± 2.17	13.13 ± 0.81
C8	2.13 ± 0.13	2.92 ± 0.40	4.65 ± 0.84
C9	10.83 ± 0.71	12.57 ± 0.76	17.52 ± 0.48
C10	5.08 ± 0.51	7.89 ± 0.47	2.37 ± 0.24
C11	1.17 ± 0.26	2.48 ± 0.67	1.47 ± 0.18
C12	7.09 ± 0.33	6.24 ± 0.31	15.72 ± 1.63
C13	3.16 ± 0.21	3.37 ± 0.53	1.81 ± 0.29
C14	17.71 ± 0.33	14.46 ± 0.72	18.43 ± 0.21
C15	6.49 ± 0.89	9.46 ± 0.21	10.87 ± 1.07
Colchicine	5.73 ± 0.26	6.25 ± 0.76	8.13 ± 0.46
Gartanin	11.68 ± 0.52	9.74 ± 0.29	7.43 ± 0.81
5-Fluorouracil	14.17 ± 2.37	17.65 ± 1.24	18.21 ± 0.87

Figure 4. Tubulin polymerisation inhibitory activity of compound C11 (A) and compound C14 (B). The concentration of colchicine was 3.5 nM.

Figure 5. Imunofluorescence staining of compound C11 against tubulin in HepG2 cells.

Figure 6. (A) Compound C11 (pale red structure) docked into a similar pocket as colchicine (blue structure); (B) 3D binging modelling between C11 and tubulin; (C) Hydrogen bonds, hydrophobic effects and hydrophilic effects of C11 targeting to tubulin (PDB code: 1SA0).

Figure 7. (A) Compound C14 (yellow structure) docked into a similar pocket as colchicine (blue structure); (B) 3D binging modelling between C14 and tubulin; (C) Hydrogen bonds, hydrophobic effects and hydrophilic effects of C14 targeting to tubulin (PDB code: 1SA0).

Figure 8. The inhibitory effects of compound C11 on NEDDylation with or without ATP.

Figure 9. (A) Compound C11 (green structure) docked into a similar pocket as B39 (red structure); (B) 3D binging modelling; (C) Hydrogen bonds, hydrophobic effects, and hydrophilic effects of C11 targeting to E1 NEDD8-activating enzyme (PDB code: 3GZN).