FIGURE 3:

Chemical derivatives of Munesib-1 increase potency against KIF15 in cells. (A) Overview of chemical derivatives of Munesib-1 that were synthesized and tested. Reagents and conditions are as follows: 1) LiOH, DMF/H2O, 25°C; 2) HATU, DIPEA, R1R2NH, DMF, 60°C; 3) TFA, CH2Cl2, RT; 4) oxalyl chloride, CH2Cl2, RT; 5) LAH, THF, 0°C, RT. (B) Quantification of the % inhibition of KIF15 microtubule gliding activity induced by each chemical derivative tested at 25 µm. Each compound was tested in singlicate, n ≥ 50 for each compound. (C) Quantification of preanaphase spindles in either monopolar or bipolar states in RPE-1 (left) or KIRC-1 (right) cells treated with each chemical derivative as well as the parent compound. Each compound was tested at 25 µM in duplicate, n = 100 for each replicate. Error bars show ± SEM. (D) CRC generated from ATPase assays with Munesib-2 (M-204) over a range of eight concentrations from 0.1 to 100. Each concentration was repeated for a total n = 4–7. Error bars show ± SEM. (E) CRC generated from microtubule gliding assays with Munesib-2 over a range of nine concentrations from 10 nM to 30 µM. Each concentration was tested in triplicate, n ≥ 50 for each replicate. Error bars show ± SEM. (F) Quantification of preanaphase spindles in either monopolar or bipolar states in RPE-1 (left) or KIRC-1 (right) cells treated with increasing concentrations of Munesib-2. Concentration is indicated on the bottom of each bar in µM. Each concentration was tested in triplicate, n = 100 for each replicate. Error bars show ± SEM.