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BRF0000320900 Begin with BRAF Searching for a target in metastatic melanoma?

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BRF0000320900 Overview Oncogenic BRAF can result from mutations in the BRAF gene, which may cause the protein to become overactive 1 One common BRAF mutation (BRAF V600 ) is implicated in diverse malignancies 2,3 Genentech is currently researching oncogenic BRAF as a novel therapeutic target 2 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. Biochem Pharmacol. 2010;80:561-567.

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BRF0000320900 Table of contents The role of RAS-RAF proteins in the RAS-RAF pathway The BRAF protein kinase Oncogenic BRAF Targeting oncogenic BRAF 3

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BRF0000320900 The role of RAS-RAF proteins in the RAS-RAF pathway 4 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. RAS BRAF ERK MEK RAF proteins play a role in the regulation of essential biologic functions 1-3 Cell growth Cell proliferation Cell differentiation

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BRF0000320900 RAS-RAF signaling in normal cells 5 1. McCubrey et al. Adv Enzyme Regul. 2006;46:249-279. Activation of the RAS-RAF signaling cascade occurs via the following sequential steps 1 : Activation by growth factors Activation of RAS Activation of RAF Phosphorylation of MEK Phosphorylation of ERK Activation of transcription factors Result 1 Cell proliferation Cell survival

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BRF0000320900 Table of contents The role of RAS-RAF proteins in the RAS-RAF pathway The BRAF protein kinase Oncogenic BRAF Targeting oncogenic BRAF 6

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BRF0000320900 The BRAF protein kinase 7 1. Wan et al. Cell. 2004;116:855-867. BRAF is a protein kinase encoded by the BRAF gene and plays an important role as an intermediary in the RAS-RAF signaling cascade 1

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BRF0000320900 Table of contents The role of RAS-RAF proteins in the RAS-RAF pathway The BRAF protein kinase Oncogenic BRAF Targeting oncogenic BRAF 8

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BRF0000320900 Oncogenic BRAF 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. V600E mutation BRAF mutations at position 600 (BRAF V600 ) can lead to overactive BRAF signaling 1 The most common BRAF mutation, BRAF V600E, is implicated in: ~50% of melanoma tumors 1 ~40% of papillary thyroid tumors 1,2 ~30% of serous ovarian tumors 2 ~10% of colorectal tumors 3 ~10% of prostate tumors 3 9

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BRF0000320900 Oncogenic BRAF signaling 10 1. Wong. Recent Pat Anticancer Drug Discov. 2009;4:28-35. 2. McCubrey et al. Adv Enzyme Regul. 2006;46:249-279. Excessive cell proliferation Resistance to apoptosis An overactive signaling cascade 1 Independent of growth factors Uncontrolled signaling Result 1,2

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BRF0000320900 Table of contents The role of RAS-RAF proteins in the RAS-RAF pathway The BRAF protein kinase Oncogenic BRAF Targeting oncogenic BRAF 11

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BRF0000320900 Research into oncogenic BRAF Research is currently ongoing to detect and target oncogenic BRAF Inhibition of oncogenic BRAF is an opportunity for precise molecular targeting 1 Research efforts are also underway to use molecular biology techniques to detect the BRAF V600 mutation 2 12 1. Wellbrock et al. Biochem Pharmacol. 2010;80:561-567. 2. Lin et al. Br J Cancer. 2011;104:464-468.

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BRF0000320900 Overview of metastatic melanoma disease state 13 1. Bhatia et al. Oncology. 2009;23:488-496. 2. Korn et al. J Clin Oncol. 2008;26:527-534. 3. McCubrey et al. Adv Enzyme Regul. 2006;46:249-279. 4. Skarin, ed. Atlas of Diagnostic Oncology. 4th ed. Philadelphia, PA: Mosby, Inc; 2010:467. Metastatic melanoma is an important area for continued research and development of strategies for patient management Median overall survival of ~8 months 1 1-year survival rate of ~25% 2 ~50% of metastatic melanoma tumors have the BRAF V600 mutation 3 Reprinted with permission from Elsevier. 4

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BRF0000320900 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 Clin Oncol. 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 V599E B-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. Biochem Pharmacol. 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. 14