Discovery, Design, and Optimization of Isoxazole Azepine BET Inhibitors School of Pharmacy, Sungkyunkwan University 2016. 6. 3. Fri. seminar. Park Sook Jin

INDEX 01 About BET inhibitor 02 Paper work - Abstract 03 Paper work – Introduction & Discussion 04 Paper work - Conclusion

Part 1 About BET inhibitor

PART1. About BET inhibitor Write emzymes : HATs, HMTs and Kinases Eraser enzymes : HDACs, DMTs, phosphatases Reader proteins : bromodomains, chromodomains, PHDs, etc. For instance, HATs deposit acetylation marks on lysine residues, which are ‘read’ by bromodomain modules and removed by HDACs.

PART1. About BET inhibitor “Bromodomains” : readers of acetyl lysine residues → The recognition of N-acetylation of lysine residues is primarily initiated by bromodomains. “Functions of bromodomains” ► Proteins that contain bromodomains are involved in the regulation of transcription - A key role in the development of cancer. ► Bromodomain-containing proteins regulate NF‑κB, which is a key transcription factor that mediates inflammatory responses. This suggests that targeting these proteins could be development of new treatment strategies for cancer, inflammation and viral infections.

PART1. About BET inhibitor ► Domain organization of BET proteins. The name and length of the proteins are shown together with the position of their first and second bromodomains. BET FAMILY (Bromodomain and Extra-Terminal) ► BRD2, BRD3, BRD4 and BRDT Each of which contains tandem bromodomains (BD1 and BD2) that recognize specific acetylated lysine residues in the N-terminal tails of histones. ★ Control expression of genes critical for proper cell growth ★ Reduction of their levels or inhibition of their activity has profound effects on cell proliferation and viability.

PART1. About BET inhibitor Structure of bromodomains ► four α-helices αZ, αA, αB and αC ► linked by diverse loop ZA loop and BC loop ★ participate in acetyllysine reader pocket formation ★ recognition elements for the modified peptide sequence ★ green triangle : approximate location of acetyllysine binding pocket.

PART1. About BET inhibitor Model of BET mechanism of action. BRD4 protein recruits positive transcription elongation factor b (P-TEFb), an essential step in RNA Pol II–mediated transcriptional elongation. A BET inhibitor blocks the binding of BRD4 to acetylated lysines on nucleosome histones, inhibiting RNA Pol II–mediated transcription.

PART1. About BET inhibitor

PART1. About BET inhibitor

PART1. About BET inhibitor BRD4 inhibition modulates MYC transcription in Acute Myeloid Leukemia. BRD4 mediates bromodomain-dependent MYC transcription, and BRD4 inhibition with JQ1 results in decreased MYC transcription in Acute Myeloid Leukemia cells. https://www.youtube.com/watch?v=Tze3XR4Kcj4

Part 2 Paper work - Abstract

Paper Work - Abstract

Paper Work - Introduction In 2009, the Mitsubishi Tanabe Pharma Corporation reported the first small molecule inhibitors of this family of chromatin adaptors, and publications followed on the use of BET inhibitors to demonstrate antitumor activity and modulate inflammation. (+)-JQ1

Paper Work - Introduction

Paper Work - Introduction Key interactions for (A) triazole 2 (PDB: 3P5O) (B) dimethylisoxazole 39 (PDB: 3ZYU) (C) pyridone-like 40 (PDB: 4E96).

Part 3 Paper work - Introduction

Paper Work - Introduction ★ Similar hydrogen bonding contacts with the asparagine(N) residue. ★ Isoxazole could serve as a replacement for the triazole. ★ 3,5-dimethyl isoxazole motif was preferred binding for bromodomains.

Paper Work - Introduction

Paper Work - Discussion

Paper Work - Introduction 3.2 Å Van der Waal distance

Paper Work - Introduction tyrosine asparagine

PART1. About BET inhibitor

Paper Work - Discussion

Paper Work - Discussion

Paper Work - Discussion

Part 4 Paper work - Conclusion

Paper Work - Conclusion ★ Compound 3 is a potent and selective BET inhibitor in biochemical and cellular assays, has a rat and dog pharmacokinetic profile suitable for in vivo experiments. ★ inhibits MYC mRNA expression in vivo after PO dosing in a dose-dependent manner. ★ Further studies detailing the optimization and development of 3 and related compounds will be reported in due course.

Paper Work - Conclusion

Paper Work - Conclusion Optimization of metabolic stability Pharmacodynamic effect in a mouse model of IL-6 suppression

Paper Work - Conclusion

Paper Work - Conclusion molecule structure OTX-015 GSK-525762 CPI-0610

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