contributions from the hydrogen bond donors and acceptors were not significant and are not shown. The binding affinities are dominated by the aromatic groups in all but one case, though both the aromatic and aliphatic groups are making favorable contribu tions to binding. Concerning the relative binding to Bcl xL versus Mcl 1, the aromatic groups selleck chem Y-27632 are leading the enhanced binding to Bcl xL in the majority of the modeling cases. These results suggest that modifica tions of the aromatic regions of JY 1 106 could be used to both improve affinity as well as alter the relative affinities for Bcl xL versus Mcl 1. JY 1 106 disrupts complex formation between Bak and anti apoptotic proteins in vitro and in tumor cells The modeling studies described above suggest that JY 1 106 binds to the anti apoptotic proteins Bcl xL and Mcl 1 in a similar fashion to that of the Bak BH3 helix.

We speculated that if JY 1 106 binds anti apoptotic proteins in this way, then it should disrupt their binding to pro apoptotic proteins. Inhibitors,Modulators,Libraries To evaluate this possibility, we first determined whether JY 1 106 disrupts the binding of Bcl xL and Mcl 1 to Bak in vitro using fluorescence polarization assays. Results show that JY 1 106 inhibits the interaction between a FITC labeled Bak BH3 peptide and Bcl xL or Mcl 1 in a dose dependent manner with IC50 values of 394 54 nM and 10. 21 0. 83 uM, respectively. The experimental Ki is about 10 times larger for Mcl 1. The results demonstrated the con current expression of both Mcl 1 and Bcl xL in most of the lines, corroborating the immunostaining results in both lung and colon tumor tissues shown in Additional file 1 Figure S1.

The cell lines were subsequently exposed to various chemotherapeutic agents at different doses, including cisplatin, SAHA, ABT 737 and JY 1 106. As demonstrated in Figure 3B, all the cancer cell lines that express relatively high levels of Bcl xL and Mcl 1, and the H23 line, which Inhibitors,Modulators,Libraries shows strong Mcl 1 expression and low Bcl xL expression, demonstrate resistance to vari ous chemotherapy agents Inhibitors,Modulators,Libraries including cisplatin, SAHA and ABT 737. Conversely, JY 1 106 causes significant tumor cell growth inhibition Inhibitors,Modulators,Libraries in these chemotherapy resistant cancer cell lines. Most interestingly, JY 1 106 is very effective in the I45 BR and DLD 1 BR cell lines, which are ABT 737 resistant cells established from parental I45 and DLD 1 cells.

To further assess whether JY 1 106 can overcome the Mcl 1 overexpression related resistance to Bcl xL inhibition, DLD 1BR and REN cells were transfected with control siRNAs or Mcl 1 siRNAs and then exposed Inhibitors,Modulators,Libraries to ABT 737. As shown in Figure 3C, after Mcl 1 reduction and ABT 737 treatment, the growth proliferation IC50 values for ABT 737 in these cells were improved to levels similar to those of JY 1 106 in untransfected cells. Given that ABT 737 is a more potent inhibitor protocol of Bcl xL in vitro than JY 1 106, these data further suggest that the superior cytotoxicity of JY 1 106 is due to its pan Bcl 2 specificity.