Yi- X iang Zhang, PhD Dana-Farber Cancer Institute

1. Potent Inhibition of Human LiposarcomaGrowth and Survival by a Novel Modulatorof MDM2-p53 Interaction Yi-Xiang Zhang, PhD Dana-Farber Cancer Institute

2. The Rationale of Targeting MDM2 in Liposarcoma MDM2/p53 Feed Back Loop WDLPS DDLPS MDM2 p53 Ubiquitination Degradation ✗ ✗ ✗ Induction of BAX, PUMA, NOXA Cell Death Induction of p21 Cell Cycle Arrest Other Biological Functions MDM2 Amplification Courtesy of Dr. Jason L. Hornick, Boston, MA Dr. Jonathan A. Fletcher, Boston, MA

3. The Rationale of Targeting MDM2 in Liposarcoma MDM2/p53 Feed Back Loop MDM2 Antagonists MDM2 p53 Induction of BAX, PUMA, NOXA Cell Death Induction of p21 Cell Cycle Arrest Other Biological Functions

4. The Development of MDM2 Antagonists for Liposarcoma Science303, 844-848 (2004) In Vivo Activation of the p53 Pathway by Small-Molecule Antagonists of MDM2 Lyubomir T. Vassilev, Binh T. Vu, Bradford Graves, Daisy Carvajal, Frank Podlaski, ZoranFilipovic, et al. Int. J. Cancer 121, 199-205 (2007) Potential for Treatment of Lipoarcomas with the MDM2 Antagonist Nutlin-3A Christoph R. Müller, Erik B. Paulsen, Paul Noordhuis, Florence Pedeutour, Gunnar Sæter, Ola Myklebost Lancet Oncol. 13, 1133-1140 (2012) Effect of the MDM2 Antagonist RG7112 on the P53 Pathway in Patients with MDM2-amplified, Well-differentiated or Dedifferentiated Liposarcoma: an exploratory proof-of-mechanism study. Isabelle Ray-Coquard, Jean-Yves Blay, Antoine Italiano, Axel Le Cesne, Nicolas Penel, JianguoZhi, et al. Here, we have established primary human liposarcoma tumor xenograft models, and evaluated efficacy of a novel MDM2 antagonist SAR299155 in vitro and in vivo.

5. Characterization of Liposarcoma Cell Lines and Primary Tumor Xenografts Cell lines: 449 and 778: Courtesy of Dr. Florence Pedeutour, Nice, France LP3 and LP6: Courtesy of Dr. Eric L. Snyder, Boston, MA LPS141, LPS510 and LPS853: Courtesy of Dr. Jonathan A. Fletcher, Boston, MA

6. SAR299155 is a Potent Inducer of p53 Activity A B C D

7. SAR299155 Decreases Cell Viability in Liposarcoma Cells with Wild-type p53 A B LP6 (p53 mut) LP6 (p53 wt) C D LP6 (+p53 siRNA)

8. SAR299155 Blocks Cell Cycle Progression and • Induces Apoptosis in Liposarcoma Cells Cell Cycle Analysis Apoptosis Analysis

9. SAR299155 Restores p53 Activity in vivo

10. Complete Regression of Primary Liposarcoma Tumor Xenograft LPS3 Treated with SAR299155 *, p<0.05; **, p<0.01; ***, p<0.001; ****, p<0.0001; compared with respective control group treated with vehicle.

11. Conclusion • SAR299155 is a potent novel MDM2 antagonist, and is 5-fold more potent than Nutlin-3 in biochemical and biological assays. • Complete and durable tumor regression were achieved in a primary human liposarcoma tumor xenograft model. • Patient tumor-derived primary liposarcomaxenograft models will be useful tools for evaluating drug efficacy and identifying new biomarkers.

12. Acknowledgement Dana-Farber Cancer Institute Andrew J. Wagner, MD, PhD EwaSicinska, MD Jeffrey T. Czaplinski Stephen P. Remillard, PhD George D. Demetri, MD Amanda L. Christie Andrew L. Kung, MD, PhD Sanofi Laurent Debussche, PhD University of Michigan Shaomeng Wang, PhD Jonathan A. Fletcher, MD Florence Pedeutour, PhD Eric L. Snyder, MD, PhD Funding Support D.K. Ludwig Fund for Cancer Research supporting the Dana-Farber/Harvard Ludwig Center Peter and Paula Fasseas Fund for Liposarcoma Research