HIF 2 HIF 2 knock out inhibits tumor growth 1,2 ARNT pVHL - PowerPoint PPT Presentation

5/4/2016 Disclosure Information PT2385: HIF ‐ 2 α Antagonist for the Treatment of Eli Wallace VHL Mutant ccRCC I have the following financial relationships to disclose: I am a stockholder and employee of Peloton Therapeutics, Inc. 12th International VHL Medical Symposium April 8, 2016 - and - I will not discuss off label use or investigational use in my presentation. Eli Wallace, Ph.D. Vice President of Chemistry Peloton Therapeutics, Inc. 1 2 April 8, 2016 HIF ‐ 2   Regulated by O 2 in Concert with pVHL HIF ‐ 2   Driver of VHL ‐ Associated ccRCC • All pVHL defective ccRCC patients express Normoxia Hypoxia HIF ‐ 2  HIF ‐ 2   O 2 O 2 Nucleus Prolyl hydroxylases Nucleus OH HIF ‐ 2  OH • Loss of VHL results in constitutive HIF ‐ 2  Defec ve VHL activation of HIF ‐  ARNT OH OH HIF ‐ 2  OH pVHL Defective VHL  HIF ‐ 2  • HIF ‐ 2  knock ‐ out inhibits tumor growth 1,2 ARNT pVHL HIF ‐ 2  ARNT • Stabilization of HIF ‐ 2  overrides tumor Angiogenesis Cell Prolifera on suppressor function of pVHL 3 pVHL & Survival OH HIF ‐ 2  • 70% of VHL disease patients develop ccRCC H y p o x OH i a ‐ R e s p o n s e E l e m e n t • ccRCC is the leading cause of mortality in VHL HIF ‐ 2  patients ARNT • Hypothesis: Inhibiting HIF ‐ 2 α will provide therapeutic benefit in the • 95% of patients with sporadic ccRCC have Cell Proliferation defective pVHL treatment of ccRCC Angiogenesis Proteasomal Degradation & Survival • Loss of VHL results in constitutive activation of 1 Kondo et al. PLoS Biology 2003, 1, 439 HIF ‐ 2  2 Zimmer et al. Mol Cancer Res 2004, 2, 89 3 Kondo et al. Cancer Cell 2002, 1, 237 3 4 April 8, 2016 April 8, 2016 1

5/4/2016 Hypoxia ‐ Inducible Factor ‐ 2  and Cancer PT2385: A Potent and Selective HIF ‐ 2  Antagonist HIF ‐  drives the expression of several genes involved in cancer HIF ‐ 2 α PT2385 Cyclin D1 VEGFA Nucleus Cell Cycle Progression Angiogenesis HIF ‐ 2  ARNT Plasminogen activator inhibitor ‐ 1 CXCR4 HIF ‐ 2 α Angiogenesis / Survival Metastasis ARNT HIF ‐ 2 α < 50 nM CD74 GLUT1 ITC Binding Affinity (K d ) Gene Immune Response Cell Metabolism HIF ‐ 1 α Inactive Hypoxia ‐ Response 786 ‐ O VEGFa ELISA (IC 50 ) 44 nM TGF  Arginase 1 Element Rat Brain:Plasma Ratio 0.9 Growth Factor Signaling Inflammatory Response HIF ‐ 2 α ARNT HIF ‐ 2  regulates the expression of multiple genes impacting • PT2385 binds to PAS ‐ B domain of HIF ‐ 2  Cell Proliferation • proliferation • Binding blocks HIF ‐ 2  :ARNT heterodimerization Angiogenesis • angiogenesis & Survival • metastasis • immune evasion 5 April 8, 2016 6 April 8, 2016 PT2385: Selective Antagonism of HIF ‐ 2  Over HIF ‐ 1  PT2385: Efficacious in 786 ‐ O Xenograft Hep3B cells (Hepatoma, VHL +/+ , HIF ‐ 1  +/+ , HIF ‐ 2  +/+ ) 786 ‐ O subcutaneous xenograft model of ccRCC ( VHL ‐ / ‐ , HIF ‐ 1  ‐ / ‐ , HIF ‐ 2  +/+ ) • Cells treated for 24 h with PT2385 at 21% O 2 (normoxia) or 1% O 2 (hypoxia) 900 800 HIF ‐ 2  specific genes HIF ‐ 1  specific genes Vehicle p.o., b.i.d. (n=8) 1% O 2 700 1% O 2 Tumor Volume (mm3) Sunitinib 40 mg/kg p.o., q.d. (n=8) 21% O 2 600 1% O 2 mean +/- SEM PT2385 3 mg/kg p.o., b.i.d. (n=8) EPO 50 PDK1 21% O 2 1% O 2 40 500 PT2385 10 mg/kg p.o., b.i.d. (n=8) Relative mRNA levels 40 Relative mRNA levels 30 400 30 20 All dose groups well tolerated 20 300 10 10 200 0 0 PT2385 (  M) PT2385 (  M) 0 0 0.1 1 10 100 0 0 0.1 1 10 0 0 5 10 15 20 25 • PT2385 inhibits expression of HIF ‐ 2  target genes with no effect on HIF ‐ 1  target genes Time After Treatment Initiation (d) • Tumor regressions with 3 and 10 mg/kg, p.o., b.i.d. dose • Similar efficacy in A498 ccRCC xenograft (VHL ‐ / ‐ , HIF ‐ 1  ‐ / ‐ , HIF ‐ 2  +/+ ) Beige SCID mice 7 8 April 8, 2016 April 8, 2016 2

5/4/2016 PT2385: Inhibits Tumor ‐ Derived VEGFA Protein Levels PT2385: Inhibition of the Expression of Multiple Genes 786 ‐ O subcutaneous xenograft model of ccRCC ( VHL ‐ / ‐ , HIF ‐ 1  ‐ / ‐ , HIF ‐ 2  +/+ ) 786 ‐ O subcutaneous xenograft model of ccRCC ( VHL ‐ / ‐ , HIF ‐ 1  ‐ / ‐ , HIF ‐ 2  +/+ ) Six doses of PT2385 p.o., b.i.d.; tumor mRNA measured 12 h post final dose Six doses of PT2385 p.o., b.i.d.; plasma protein VEGFA measured 12 h post final dose 300 1.5 1.5 Cyclin D1 VEGFA Relative mRNA levels Relative mRNA levels hVEGFA (pg/mL) mean +/- SEM 200 1.0 1.0 0.5 0.5 100 0.0 0.0 PT2385 (mg/kg) PT2385 (mg/kg) 0 1 3 10 0 1 3 10 0 2.0 1.5 PAI-1 GLUT1 Relative mRNA levels Relative mRNA levels Vehicle 1 mg/kg 3 mg/kg 10 mg/kg 1.5 PT2385 PT2385 PT2385 1.0 1.0 Treatment Group 0.5 0.5 • PT2385 has no effect on mouse VEGFA levels (data not shown) 0.0 0.0 PT2385 (mg/kg) 0 1 3 10 0 1 3 10 PT2385 (mg/kg) • PT2385 inhibits mouse kidney EPO gene expression (data not shown) • Dose ‐ dependent Inhibition of HIF ‐ 2  regulated genes Beige SCID mice Beige SCID mice 9 April 8, 2016 10 April 8, 2016 PT2385: Inhibits Proliferation and Angiogenesis PT2385: Efficacious in Sunitinib Refractory PDX Model 786 ‐ O subcutaneous xenograft model of ccRCC ( VHL ‐ / ‐ , HIF ‐ 1  ‐ / ‐ , HIF ‐ 2  +/+ ) PDX subcutaneous xenograft model of ccRCC ( VHL ‐ / ‐ , HIF ‐ 1  +/+ , HIF ‐ 2  +/+ ) Six doses of PT2385 10 mg/kg p.o., b.i.d.; tumor tissue collected 12 h post final dose • Model derived from a patient refractory to both sunitinib and everolimus CD ‐ 31 Ki67 1000 Vehicle p.o., b.i.d. 800 Tumor Volume (mm 3 ) Sunitinib 40 mg/kg, p.o., q.d. Vehicle mean +/- SEM PT2385 30 mg/kg, p.o., b.i.d. 600 400 All dose groups well tolerated 200 PT2385 0 0 5 10 15 20 25 Time After Treatment Initiation (d) • PT2385 efficacious in sunitinib refractory model • Tumor expresses both HIF ‐ 1  and HIF ‐ 2  • PT2385 treatment reduces proliferation (Ki67) and angiogenesis (CD ‐ 31) CRO: Champions Oncology 11 12 April 8, 2016 April 8, 2016 3

5/4/2016 Effect of PT2385 and Sunitinib on Blood Pressure Effect of PT2385 and Sunitinib on Blood Pressure VEGFR TKIs and VEGFA mAb are associated with hypertension in patients VEGFR TKIs and VEGFA mAb are associated with hypertension in patients The effect of sunitinib on blood pressure and heart rate are recapitulated in rats The effect of sunitinib on blood pressure and heart rate are recapitulated in rats 150 150 Vehicle p.o., b.i.d. (n=3) Vehicle p.o., b.i.d. (n=3) Mean Arterial Pressure (mm Hg) Mean Arterial Pressure (mm Hg) Sunitinib 40 mg/kg p.o., q.d. (n=3) Sunitinib 40 mg/kg p.o., q.d. (n=3) PT2385 30 mg/kg p.o., b.i.d. (n=3) PT2385 100 mg/kg p.o., b.i.d. (n=3) 125 125 100 100 75 75 0 25 50 75 100 125 0 25 50 75 100 125 telemeterized SD rats telemeterized SD rats Time (hours) Time (hours) • PT2385 had no effect on blood pressure at 5x the maximal efficacious exposure • Sunitinib significantly increased blood pressure at its maximally efficacious dose • PT2385 had no effect on heart rate or any other ECG parameter • Sunitinib also significantly reduced (10 ‐ 20%) heart rate • PT2385 has a favorable preclinical safety profile CRO: CorDynamics CRO: CorDynamics 13 April 8, 2016 14 April 8, 2016 PT2385 – 101 PT2385 ‐ 101: Pharmacodynamic Response in Patients Objectives & Design Primary Objective: Erythropoietin expression is regulated by HIF ‐ 2 α • To identify the maximum tolerated dose (MTD) and/or the recommended Phase 2 dose of PT2385 tablets in patients with advanced clear cell renal cell carcinoma (ccRCC) Average %Change in Erythropoietin 40 Secondary Objectives: 20 % Change in Erythropoietin • To evaluate the safety profile of PT2385 0 (Average +/- SD) • To determine the pharmacokinetic (PK) profile of PT2385 dose level 1 (n=3) dose level 2 (n=3) • To assess the pharmacodynamic (PD) effects of treatment with PT2385 -20 dose level 3 (n=4) • To assess the anti ‐ tumor activity of PT2385 -40 dose level 4 (n=7) dose level 5 (n=6) -60 Design: dose level 6 (n=3) -80 • 3+3 Design -100 • Dose limiting toxicity (DLT) observation period 3 weeks 0.0 0.5 1 8 Day • Expansion to 25 additional patients at MTD or RP2D • In patients, target engagement, as measured by diminution of erythropoietin expression, is exposure related, rapid and pronounced 15 16 April 8, 2016 April 8, 2016 4