Begin with BRAF

1. Begin with BRAF Searching for a target in metastatic melanoma?

2. Overview • Oncogenic BRAF can result from mutations in the BRAF gene, which may cause the protein to become overactive1 • One common BRAF mutation (BRAFV600) is implicatedin diverse malignancies2,3 • Genentech is currently researching oncogenic BRAF as a novel therapeutic target 1. Sharma et al. Cancer Res. 2005;65:2412-2421. 2. Wong. Recent Pat Anticancer Drug Discov. 2009;4:28-35. 3. Wellbrock et al. BiochemPharmacol. 2010;80:561-567.

3. Table of contents The role of RAS-RAF proteins in the RAS-RAF pathway The BRAF protein kinase Oncogenic BRAF Targeting oncogenic BRAF

4. The role of RAS-RAF proteins in the RAS-RAF pathway • RAF proteins play a role in the regulation of essential biologic functions1-3 • Cell growth • Cell proliferation • Cell differentiation • RAS • BRAF • MEK • ERK 1. Wong. Recent Pat Anticancer Drug Discov. 2009;4:28-35. 2. Wan et al. Cell. 2004;116:855-867. 3. McCubrey et al. Adv Enzyme Regul. 2006;46:249-279.

5. RAS-RAF signaling in normal cells Activation of the RAS-RAF signaling cascade occurs via the following sequential steps1: • Activation by growth factors • Activation of RAS • Activation of RAF • Phosphorylation of MEK • Phosphorylation of ERK • Activation of transcription factors Result1 • Cell proliferation • Cell survival 1. McCubrey et al. AdvEnzyme Regul. 2006;46:249-279.

6. Table of contents The role of RAS-RAF proteins in the RAS-RAF pathway The BRAF protein kinase Oncogenic BRAF Targeting oncogenic BRAF

7. The BRAF protein kinase BRAF is a protein kinase encoded by the BRAF gene and plays an important role as an intermediary in the RAS-RAF signaling cascade1 1. Wan et al. Cell. 2004;116:855-867.

8. Table of contents The role of RAS-RAF proteins in the RAS-RAF pathway The BRAF protein kinase Oncogenic BRAF Targeting oncogenic BRAF

9. Oncogenic BRAF • BRAF mutations at position 600 (BRAFV600) can lead to overactive BRAF signaling1 • The most common BRAF mutation, BRAFV600E, is implicated in: • ~50% of melanoma tumors1 • ~40% of papillary thyroid tumors1,2 • ~30% of serous ovarian tumors2 • ~10% of colorectal tumors3 • ~10% of prostate tumors3 • V600E mutation 1. McCubrey et al. Adv Enzyme Regul. 2006;46:249-279. 2. Pritchard et al. Biochem Soc Trans. 2007;35:1329-1333. 3. Cho et al. Int J Cancer. 2006;119:1858-1862.

10. Oncogenic BRAF signaling An overactive signaling cascade1 • Independent of growth factors • Uncontrolled signaling Result1,2 Excessive cell proliferation Resistance to apoptosis 1. Wong. Recent Pat Anticancer Drug Discov. 2009;4:28-35. 2. McCubrey et al. Adv Enzyme Regul. 2006;46:249-279.

11. Table of contents The role of RAS-RAF proteins in the RAS-RAF pathway The BRAF protein kinase Oncogenic BRAF Targeting oncogenic BRAF

12. Research into oncogenic BRAF • Research is currently ongoing to detect and target oncogenic BRAF • Inhibition of oncogenic BRAF is an opportunity for precise molecular targeting1 • Research efforts are also underway to use molecular biology techniques to detect the BRAFV600 mutation2 1.Wellbrock et al. BiochemPharmacol. 2010;80:561-567. 2. Lin et al. Br J Cancer. 2011;104:464-468.

13. Overview of metastatic melanoma disease state • Median overall survival of ~8 months1 • 1-year survival rate of ~25%2 • ~50% of metastatic melanoma tumors have the BRAFV600 mutation3 Reprinted with permission from Elsevier.4 Metastatic melanoma is an important area for continued research and development of strategies for patient management 1. Bhatia et al. Oncology. 2009;23:488-496. 2. Korn et al. J ClinOncol. 2008;26:527-534. 3. McCubrey et al. AdvEnzyme Regul. 2006;46:249-279.4.Skarin, ed. Atlas of Diagnostic Oncology. 4th ed. Philadelphia, PA: Mosby, Inc; 2010:467.

14. References Bhatia S, Tykodi SS, Thompson JA. Treatment of metastatic melanoma: an overview. Oncology. 2009;23:488-496. Cho NY, Choi M, Kim BH, Cho YM, Moon KC, Kang GH. BRAF and KRAS mutations in prostatic adenocarcinoma. Int J Cancer. 2006;119:1858-1862. Korn EL, Liu PY, Lee SJ, et al. Meta-analysis of Phase II cooperative group trials in metastatic stage IV melanoma to determine progression-free and overall survival benchmarks for future Phase II trials. J ClinOncol. 2008;26:527-534. Lin J, Goto Y, Murata H, et al. Polyclonality of BRAF mutations in primary melanoma and the selection of mutant alleles during progression. Br J Cancer. 2011;104:464-468. McCubrey JA, Steelman LS, Abrams SL, et al. Roles of the RAF/MEK/ERK and PI3K/PTEN/AKT pathways in malignant transformation and drug resistance. Adv Enzyme Regul. 2006;46:249-279. Pritchard C, Carragher L, Aldridge V, et al. Mouse models for BRAF-induced cancers. Biochem Soc Trans. 2007;35:1329-1333. Sharma A, Trivedi NR, Zimmerman MA, Tuveson DA, Smith CD, Robertson GP. Mutant V599EB-raf regulates growth and vascular development of malignant melanoma tumors. Cancer Res. 2005;65:2412-2421. Skarin AT, ed. Atlas of Diagnostic Oncology. 4th ed. Philadelphia, PA: Mosby, Inc; 2010:467. Wan PT, Garnett MJ, Roe SM, et al. Mechanism of activation of the RAF-ERK signaling pathway by oncogenic mutations of B-RAF. Cell. 2004;116:855-867. Wellbrock C, Hurlstone A. BRAF as therapeutic target in melanoma. BiochemPharmacol. 2010;80:561-567. Wong KK. Recent developments in anti-cancer agents targeting the Ras/Raf/MEK/ERK pathway. Recent Pat Anticancer Drug Discov. 2009;4:28-35.