1. ----- Identification of Potent SHP2 inhibitors Liwei Chen Ph.D. Dr. Jerry Wu Lab Discovery of Novel SHP2 inhibitors

2. SHP2 Protein Tyrosine Phosphatase Cys

3. Germline mutations: Noonan Syndrome, leukemias Somatic mutations: Myeloid malignancies, solid tumors Known SHP2 mutations in human cancer Cancer Res. (2004) 64: 8816-8820

5. Deregulation of SHP2 by CagA

6. The links between activating mutations in SHP2 and cancer suggest that SHP2 may be the product of an oncogene and leukemia-associated Shp2 mutants be attractive therapeutic targets

9. Flowchart of the purification strategy GST-Shp2 expressed in E.coli Eluate used for PTP assays GSH Sepharose Elute with gluthatione

10. Screening Strategy of the NCI Diversity Set NCI diversity set (1990 compounds) Screen (at 10  M) DiFMUP-based PTPase assay Confirm primary hits 33 active compounds (50% inhibition) Obtain active compounds from NCI Phosphopeptide-based PTPase assay (at 10  M) 24 active compounds (50% inhibition) Determine IC50 for SHP2 and SHP1 PTPases

11. Summary of IC50 of potential inhibitors >30 (~50) 5.034 87877 no inhibition 3.243 69343 13.15 3.238 117199 17.45 3.19 50352 5.506 0.332 339601 2.693 0.303 119910 0.737 0.19 119913 Rodent SHP1 Rodent SHP2 IC50 (  M) NSC ID

12. Molecular model of NSC-87877 binding to the SHP2 PTP domain

13. NSC-87877 docked to SHP2

14. NSC-87877 docked to SHP1

15. Recognition loop

17. N, number of experiments. Each experiment was performed in triplicates. Table I. Inhibition of PTPs by NSC-87877 in vitro PTP IC50 Selectivity (fold) SHP2 0.35 + 0.15  M (N = 9) 1 SHP1 0.35 + 0.16  M (N = 5) 1 PTP1B 1.69 + 0.81  M (N = 4) 5 HePTP 7.73 + 3.12  M (N = 4) 22 CD45 84.68 + 28.03  M (N = 3) 242 hLAR-D1 150.93 ± 18.15  M (N = 4) 431 hDEP1 65.62 ± 7.14  M (N = 3) 187 SHP2V280 0.99 + 0.35  M (N = 4) 3 SHP2RD 1.01 + 0.49  M (N = 4) 3

20. Model outlining the role of Gab1 and SHP2 in EGF-induced Ras activation Growth factor, cytokines RTK or JAK associated receptor Gab1/Gab2 Sos-Grb2 SHP2 Ras Ren Y,. Et al., J. Biol. Chem., Vol. 279, Issue 9, 8497-8505, February 27, 2004

21. NSC-87877 inhibits the SHP2 PTP activity in cells

29. A B SHP2E76K activates Ras-Erk pathway in TF-1 human myeloid leukemia cells

31. SHP2E76K activates Src family kinases (SFKs) A B

32. SHP2E76K sustains TF-1 cells cytokine-independent growth and hypersensitivity to GM-CSF A B C D

38. Acknowledgement Dr. Jerry Wu Lab Yuan Ren, Ph.D. Zhengming Chen, Ph.D. Hai Wang, Ph.D. High-through Screening Facility Core Richard Yip Shen-Shu Sung Wayne Guida Drug Discovery Program Harshani Lawrence Nicholas J. Lawrence Said M. Sebti