Novel Transcription Factor Inhibitor as Treatment for Epithelial Cell Cancers John Bushweller, Department of Molecular Physiology and Biological Physics, University of Virginia Jeff James Licensing Manager UVA Licensing and Ventures Group

Value position Novel small molecule inhibitors that are potent against breast, ovarian, and lung epithelial cancer cell lines and use a les exploited mechanism by targeting a transcription factor involved in cancer pathways Transcription factor inhibition as treatment mechanism Potent against epithelial cancer cell lines in vitro Specific ablation of cancer cell and not normal cell lines in vitro Potential for targeting of cancer stem cells Compounds are orally bioavailability Potentially low toxicity Compounds target multiple cancer types

Market Respiratory Cancers: 67% mortality in US Market expected to grow from $4 billion in 2010 to $13.5 billion by 2020 Breast Cancers: 18% mortality in US Market expected to grow from $9.8 billion in 2013 to $18.2 billion by 2023 Ovarian Cancers: 67% mortality in US Market expected to grow from $460 million in 2011 to $1.4 billion by 2021 Current treatments utilize monoclonal antibodies or small molecule inhibitors Pitfalls: Off target effects, toxicity, moderately effective, most converge on small number of biological pathways and downstream effectors, e.g. EGFR, Ras, PI3K, Her2 There is the need for more potent therapies that address other pertinent pathways involved in breast, ovarian and lung cancer progression Source: World Cancer Research Fund, American Cancer Society, IMShealth, Markets&Markets, Decision Resources

Mechanism of action CBFβ and RUNX1 are proteins that together to form Core Binding Factor, a transcription factor involved in cellular development Compounds bind to CBFβ and block its interaction with RUNX proteins, abrogating downstream signaling events associated with cancer

Compounds Kill Epithelial Lung Cancer Cells but Not Normal Lung Cells A) Active inhibitor reduces lung cancer cell viability to similar degree as staurosporine, a well-defined inducer of cell death, compared to treatment with DMSO and inactive control. B) Active inhibitor has no deleterious effect on viability of normal lung cell line as compared to staurosporine treatment. B) AI-10-104 Active inhibitor AI-4-88 Inactive control

CBFβ Inhibitor Ablates Viability of Ovarian Cancer Cell Lines Dose-dependent effect of AI-10-104 inhibitor on ovarian cancer cell lines after 72 hours

Compounds are Effective Against Breast Cancer Cell lines in 3D Organotypic Culture A) Brightfield microscopy of colony breast cancer cell line colony forming units (CFU) in 3D culture B) Enumerated CFUs show active inhibitor reduces growth of cell line in 3D culture compared to control treatments. A) B) Active inhibitor (AI-14-91, AI-10-104) 1 µM Inactive control (AI-4-88) 1 µM AI-14-91 is a derivative of AI-10-104 with improved in vivo properties

CBFβ Inhibitors are Orally Bioavailable : Plasma concentration as a function of time for active inhibitors AI-12-126 or AI-14-91 : concentration from oral gavage dosing  : Concentration from intraperitoneal dosing (100 mg/kg) in mice.   AI-12-126 Intraperitoneal AI-14-91 Oral Gavage AI-14-91 Intraperitoneal Half life (min) 179 203 243 AUC(0-inf) (µmol∙min/L) 8.14 2.22 2.08 Mean Residence Time (min) 74.6 180.4 90.6 Volume of Distribution (mL) 193.9 754.0 958.7 Clearance (mL/min) 0.75 2.57 2.74

Future Work We’re continuing development of the compounds: Testing inhibitor efficacy in vivo, 6-8 months Optimizing compounds – 6-12 months Test toxicity of optimized compounds in vivo, 12-18 months UVa is currently searching for partners to continue developing these compounds as anti- tumor agents

UVa Licensing and Ventures Group Contact: Jeffrey A. James, Ph.D., CLP. Licensing Manager UVa Licensing and Ventures Group 722 Preston Ave., Ste. 107 Charlottesville, VA 22902 (434) 982-1608 (direct) (434) 924-2175 (main) (434) 982-1583 (fax) jeff.james@virginia.edu www.innovation.virginia.edu