1. Screening of allosteric inhibitors and inhibitors of protein-protein interactions Joel Eggert

2. Kinase inhibitors Orthosteric Foda & Seeliger, 2014.

3. Kinase inhibitors OrthostericAllosteric Foda & Seeliger, 2014.

4. Allosteric inhibitors Advantages – Highly specific – Lower chances of side effects – Overcome clinically acquired resistance – Barely explored

5. Allosteric inhibitors Disadvantages – Allosteric sites are often unknown – Hard to predict the outcome – Higher divergence between species

6. High-throughput screen of allosteric inhibitors Goal: identify targets that can stabilize the enzyme in a catalytically inactive conformation

7. Recombinant cSrc Site-directed mutagenesis + Cys activation loop Acrylodan O/N Mass spec Triplicates: acrylodan-cSrc (100 nM) Compounds: 10 and 50 µM 15-30 min, RT

8. Simard et al., 2009

9. inhibitorbindingkinase Type Iactive siteactive state Type IIactive + allosteric siteinactive state Type IIIallosteric siteinactive state Simard et al., 2009

10. inhibitorbindingkinase Type Iactive siteactive state Type IIactive + allosteric siteinactive state Type IIIallosteric siteinactive state Simard et al., 2009

11. inhibitorbindingkinase Type Iactive siteactive state Type IIactive + allosteric siteinactive state Type IIIallosteric siteinactive state Simard et al., 2009

13. Inhibitors of protein-protein interactions

14. http://www.singerinstruments.com/application/yeast-2-hybrid

15. Chemical library: >156 000 compounds (300-600 kDa)

16. Fragment-screening and fragment- based drug design (FBDD) Low molecular weight compounds NMR Bind – target protein? Weak interactions [mM] Fragment hits Structure-based optimization (fragment linking etc) Lead compound

17. Sheng et al., 2015

19. References Foda, Z. H. & Seeliger, M. A. Kinase inhibitors: An allosteric add-on. Nat. Chem. Biol. 10, 796–797 (2014). Nussinov, R. & Tsai, C.-J. Allostery in disease and in drug discovery. Cell 153, 293–305 (2013). Hajduk, P. J. & Greer, J. A decade of fragment-based drug design: strategic advances and lessons learned. Nat. Rev. Drug Discov. 6, 211–219 (2007). Simard, J. R. et al. A new screening assay for allosteric inhibitors of cSrc. Nat. Chem. Biol. 5, 394–396 (2009). Sheng, C., Dong, G., Miao, Z., Zhang, W. & Wang, W. State-of-the-art strategies for targeting protein-protein interactions by small-molecule inhibitors. Chem. Soc. Rev. 44, 8238–8259 (2015).