‘COMMERCIAL FEASIBILITY OF THE MULTICOMPONENT NANOCHAIN THERAPY’ By CHAITANYA GOLLAKOTA Submitted in partial fulfillment of the requirements For the degree of Masters of Science Thesis Adviser: Dr. Christopher Cullis DEPARTMENT OF BIOLOGY CASE WESTERN RESERVE UNIVERSITY AUGUST 2016 CASE WESTERN RESERVE UNIVERSITY SCHOOL OF GRADUATE STUDIES We hereby approve the thesis/dissertation of Chaitanya Gollakota candidate for Master of Science (Signed) Ed Caner Dr. Christopher Cullis Dr. Emmit Jolly Dr. Roy Ritzmann (Date) 07/06/2016 Acknowledgements My thesis project and my graduate degree program would have been a lot less exciting and stressful without the following people: My heartfelt thanks to Drs. Efstathios Karathanasis and Mark Griswold for letting me work on their area of research. Special thanks to Dr. Karathanasis for pushing me to the limit during the development of my thesis. A huge thanks to Drs. Christopher Cullis and Emmit Jolly for instilling in me a lot of confidence during the course of my graduate degree program, and during the development of my thesis. Thank you Dr. Cullis, who was my thesis advisor, for being patient with me. I cannot thank Ed Caner and Dr. Roy Ritzmann enough for joining my thesis defense committee with a very limited notice of time. A special thanks to Bruce Terry and the rest of the STEP program faculty for all the fantastic mentorship I received. The following people from the industry helped me shape my thesis by giving wonderful insights • Suzette Dutch - Triathlon Medical Ventures • Neema Mayhugh - Wave Strategy LLC • Dr. Andrew Sloan - University Hospitals • Nicole Brey - Cleveland Clinic • Drs. Saunthararajah, M.D. Yogen and Velcheti, M.D. Vamsidhar - Cleveland Clinic • Raghuram Selvaraju - Rodman & Renshaw And last but never the least, my parents - Jayashree Gollakota and Satyanarayana Gollakota. I don’t think any of this would have been possible without their support. Special Thanks - James Brown. Your songs made my lonely days of writing this thesis much better. Contents Technology Highlights 1 Product Overview 6 Metastatic Breast Cancer 6 Matrix Metalloproteinase (MMPs) 12 Loss of Cell Adhesion is a Crucial Factor in Metastatic Cancers 13 Tumor Angiogenesis: 14 Epithelial-Mesenchymal Transition (EMT) 15 Metastatic Microenvironment 16 Organ Selection 17 Tiny Particles 18 Introduction to Nanotechnology 18 Evolution of Nanotechnology 19 First Generation Nanoparticles 20 Adagen 20 Zoladex 20 Interaction between the Complement System and the Nanoparticles 21 Second Generation Nanoparticles: 22 Doxil 23 Lipodex 24 Myocet 24 Third Generation Nanoparticles 25 Limitation with the EPR Effect 25 Factors Affecting the Uptake of NPs into Cancer Cells 26 Size and Charge 26 Encapsulating Liposome 27 Nanochains 27 Targeting Ligand 28 Role of Integrins in Metastasis: 29 TECHNOLOGY LANDSCAPE 30 Issues with Targeting Integrins/ Associated Ligands 30 Number of Receptors on the Cancer Cells vs. Normal Cells 30 Externally Modulated Release Mechanism 30 Magnetic Field Modulation: 32 ThermaDox 33 Results of Nanochain 34 Shape of the Nanoparticle: Extremely Crucial Factor 34 RGD Ligand Targeting 35 RGD Ligand Targeting causes Inflammation 36 Single Therapy vs. Combination Therapy 37 Advantages of a Combination Therapy 37 Designing Drug Combinations 38 Surface Modification of Nanoparticles 38 Circulation Time 39 Surface Charge 39 Clinical Trial Strategy 40 Market 42 Jobs To Be Done 43 Product Evolution 43 Cost 43 Hospital Costs 44 Function 45 Convenience 45 Reliability 45 Cancer Survival 46 Research Strategies 46 Co-Innovators 46 Incremental Innovation 47 Nanochain: Benefits to Costs 48 Competition 49 Glucose Inhibitors 49 p53 Pathway Activation 50 Monoclonal Antibodies (mAbs) 52 Market: 53 A Few Promising Products: 54 Keytruda: PD-1: Major Threat: Promising against TNMBC 55 Nanochain’s Opportunity 58 Pembrolizumab: Disease Indications 61 Avastin: 61 Liposomal Particles 64 BBB Therapeutics 64 Mebiopharm 65 Polymer-based Therapeutics 66 Engene IC 67 Immunoliposomes 69 Popular methods of making liposomes: 71 Anti-EGFR 75 Anti-RON Receptor Tyrosine Kinase: 76 Future Threats and Recommendations 79 A case of targeting multiple integrin-mediated pathways 79 Efficacy: 81 Competition in the Triple Negative Metastatic Breast Cancer Market 82 Problem with Metastatic Cancers: Multidrug resistance 86 Severity of the Disease: 87 Approved Therapies for TNBC: 89 Appendix 93 References 107 List of Tables: Circulation times for various nanoparticles used in the nanochain therapy 39 Results of Glufosfamide 49 Results of Synergene’s Product 52 Results of an anti-EGFR-ILs-DOX developed by the University Hospital Basel 55 Results of Keytruda’s performance against Triple negative metastatic breast cancers 56 Results of 2B3-101 65 Results of MBP-426 66 Results of BIND’s trial 67 Results of Engene IC 68 Results of Intetuamab 79 Results from a Combination Therapy Trial 86 List of Figures Missing receptors on triple negative breast cancer cells 2 Pipeline of Nanochain Therapy 3 Five-year Relative Survival Rates* (%) by Stage at Diagnosis, US, 2005-2011 4 Ras oncogene function in tumor metastasis 7 Son of Sevenless Pathway 9 Ras Protein Downstream Pathway 10 Rho Family of Proteins 11 Loss of Cell Adhesion a Major Cause for Metastasis 13 Increase in the number of nanoparticles developed during the past decade 19 Source: NIH Reporter 19 1 - Shows the nanoparticle engulfed by the phagocyte 2 - Shows a coated nanoparticle reaching its target cell 22 EPR Effect 23 Doxil 24 RGD amino acid sequence 28 Integrin pathway in tumor development 29 RGD amino acid binding sites 37 Structure of a positively charged nanoparticle, 40 Expired Patent of Docetaxel 47 A schematic representing the role of the p53 pathway in tumor invasion 51 Kaplan Meier Statistical Analysis of the Clinical Trial 60 Immunoliposome sketch 69 Receptor-mediated endocytosis 70 Various types of Immunoliposomal formulation 72 Commercial Feasibility of the Multicomponent Nanochain Therapy Abstract by CHAITANYA GOLLAKOTA Abstract: One of the most dangerous forms of cancers are metastatic cancers for which there is no comprehensive treatment regimen. Drs. Karathanasis and Griswold at Case Western Reserve University are developing a targeted cancer therapy called nanochain that accumulates exclusively near metastatic tumor sites delivering docetaxel and doxorubicin. In a recent animal model, the anti-cancer agent demonstrated close to 6% accumulation rates in an animal model. This thesis conducts a comprehensive analysis on the commercial feasibility of the nanochain product and comes to a conclusion that the product needs to prove that it can improve the overall survivability of metastatic patients compared to its competition like monoclonal antibodies and kinase pathway inhibitors. This thesis suggests that it will be a good commercial strategy to conjugate the liposomal portion of the nanochain with a novel molecule like a monoclonal antibody (mAb). Technology Highlights • Market Area: According to the American Cancer Institute, close to 15 million Americans were living with a history of cancer as of January 1, 2014. In the year 2016, about 1.7 million new cancer cases are expected to be diagnosed. What is even more dangerous is the fact that close to 600,000 Americans are projected to die of cancer in the year 2016. Cancer follows cardiac-related diseases in the number of fatalities each year. There is an unmet need to treat tumor before it takes the lives of more people. A drug delivery platform developed by biomedical researchers at Case Western Reserve University claiming to have a higher therapeutic index than therapeutic agents of this class Drs. Efstathios Karathanasis and Mark Griswold are fighting a compelling battle against cancer using % of Cancer Deaths by Various Cancer Types in the Year 2016 a multicomponent Source: American Cancer Society 2% 3% 9% 6% nanoparticle called Breast Cancer 2% 0% 4% Nanochain. One of 1% Colorectal cancer 34% 15% 5% the therapeutic areas 5% Kidney Cancer 9% of focus is triple 6% 3% 11% negative metastatic breast cancer, a form of cancer without any comprehensive treatment. Approximately 40,000 deaths are expected to occur in the year 2016 due to triple negative breast cancer indicating that there is a need to develop an anti-cancer agent for this orphan disease class. • The name ‘triple negative’ comes from the fact that these forms of tumor cells do not have receptors for estrogen, progesterone, and Human Epidermal Growth Factor 1 Receptor 2(HER2). Developing drugs against this type of cancer has been a challenge to the scientific community due to the absence of these receptors. Herceptin is a triple positive metastatic breast tumor-targeting monoclonal antibody. Trastuzumab, the drug’s generic name, accumulated close to $7 billion in the year 2013, targeting all the three receptors overexpressed on cancer cells - Her2, estrogen, and progesterone. Figure: Missing receptors on triple negative breast cancer cells Estrogen Progesterone HER2 2 Nanochain: A Pipeline in a Single Product: The product comprises of three iron oxide particles, a liposome, and a targeting entity on the surface. Nanochain targets a severe unmet need by accumulating precisely close to triple negative metastatic breast cancer sites inside the body. The product is also being tested for glioblastoma multiforme in a recent preclinical trial. The market for this therapeutic area is expected to grow by more than 10% over the next years and is predicted to reach $1 billion