STATE OF GEORGIA COUNTY OF CLARKE Affidavit of Dr. Jason Zastre ‘Comes now the Affiant, Dr. Jason Zastre, who being first duly swom by an officer authorized by law to administer oaths, states the following: 1 My name is Dr. Jason Zastre, I am over the age of 18, and I am a tenured Associate Professor in the Department of Pharmaceutical & Biomedical Sciences at the University of Georgia, College of Pharmacy. I have been a member of the Graduate Faculty at the University of Georgia since 2007. I received my Bachelors of Science in Pharmacy in 1994 and my Masters of Science in Pharmaceutical Sciences in 1998 from the University of Manitoba in Winnipeg, MB. Canada. In 2005 I received my Ph.D. in Pharmaceutics and Biopharmaceutics from the University of British Columbia, Vancouver, BC. Canada. 2. In my work at the University of Georgia, 50% of my time is devoted to teaching and 50% to research. I teach graduate level and undergraduate level students. In the Doctorate of Pharmacy program, I am the Course Coordinator and responsible for all material in Pharmaceutics I: Physical Pharmacy and Dosage Form Design. A true and correct copy of my Curriculum Vitae is attached as Exhibit A. a Twas contacted by attomey Sabrina Graham of the Georgia Attomey General’s Office and Robert Jones, General Counsel for the Georgia Department of Corrections and asked to provide assistance to the State of Georgia in evaluating what occurred in a sample of compounded pentobarbital sodium solution which apparently precipitated after shipment on frozen gel packs and storage at approximately 37 degrees Fahrenheit over more than 7 days. 4. After viewing a video of the solution and learning about the shipment and storage of the solution, my assessment of the formulation indicates that pentobarbital had precipitated or fallen out of solution. In order to confirm the identity of the precipitated particles within the solution, I recommended that a sample of the solution and a sample of the powder wihich was used to prepare the solution be sent for analysis to a sophisticated testing facility to evaluate the particulate matter in the solution and then compare it to the powder used to prepare the solution. located a testing laboratory which was equipped to perform this work, Triclinic Labs, located in Lafayette, Indiana,‘The Department of Corrections provided Triclinic Labs with one pentobarbital powder sample ‘and one syringe containing a solution with suspended solids. Each of these samples was analyzed using x-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and infrared (IR) spectroscopy. A true and correct copy of the Triclinic Labs testing report, which describes the analysis in detail, is attached as Exhibit B. 6. ‘The Triclinic Labs report contains the following conclusions: Based on the analysis performed in this study, the powder sample is believed to contain pentobarbital sodium, a polymorph of pentobarbital free base, and sodium chloride. The solids isolated from the syringe are believed to contain two different polymorphs of pentobarbital free base. (See Exhibit B, p. 4). 7. Based on the Triclinic Labs analysis, the solid materials in the solution were identified to be the active pharmaceutical ingredient, pentobarbital, but in two different solid states. Essentially, the “polymorph” referenced in the Triclinic Report, refers to a different crystalline form of the same pure substance in which the molecules have different arrangements. A rudimentary example of this type of solid state conversion is carbon forming either a diamond crystal or graphite; both are still solid carbon just in a different solid states with different physical properties. & In this case, the Triclinic testing report confirms that the pentobarbital sodium solution precipitated, and the solid particles remaining in the solution were two different solid forms of pentobarbital. - ‘There is no evidence that the solution was adulterated. Instead, the analysis performed by Triclinic Labs indicates that the particles within the solution were simply a polymorph of pentobarbital, which precipitated or fell out of solution. 10. ‘The physical state (e.g. gas, liquid, solid), of a molecule of a particular drug is not an absolute property, but instead dependent upon the conditions under which itis placed. A different solid state, such as a polymorph, can occur for a number of reasons, including the temperature at which the substance is stored, the rate of cooling of the substance, or in the case of a liquid pharmaceutical formulation, slight variations in the concentration of a pharmaceutical solvent used to dissolve the drug into the liquid state.UL In this case, the most likely cause of the precipitation observed within the solution was that the solution was shipped and stored at a temperature which was too low. An additional possible cause could be if the pharmaceutical solvent used to dissolve the pentobarbital sodium had absorbed some amount of water or evaporated during the preparation process. This may result in a lower concentration of solvent, ultimately impacting the solubility of the drug, which increases the possibility of precipitation. 12. ‘The manufacturer of the commercially available injectable form of pentobarbital sodium advises. that the solution should be stored at a controlled room temperature, not less than 59 degrees Fahrenheit. The reason for this is that the concentrated form of pentobarbital sodium solution is most likely close to the solubility limit, so the drug may precipitate or fall out of solution at lower temperatures. 13. ‘The State of Georgia can minimize the possibility of precipitation within the solution by storing the solution at a controlled room temperature above 59 degrees Fahrenheit, and by assuring that the pharmacist preparing the solution takes steps to minimize the possibility that the pharmaceutical solvent evaporates or absorbs water during the pharmaceutical compounding. process. FURTHER AFFIANT SAYETH NOT Executed this iS day of. foc | 2015. 4 Jason Zasttel Hb.D. ‘Swom and subscribed before me, this [S3h day of, Bepad , 2015. Notary Pupte 6 ‘My commission expires eng B0Lr i Seog/ “aaa iwEXHIBIT ACurriculum Vitae 1. Academic History a, Name: b. Present rank: Jason Allan Zastre Associate Professor with Tenure Department of Pharmaceutical & Biomedical Sciences College of Pharmacy, University of Georgia 250 West Green Street, Room 222 Athens, GA 30602 Phone: (706) 583-0290 Mobile: (706) 296-2399 E-mail: jzastre@rx.uga.edu ©. Proportion time assignments: _0.75 EFT (9-month); 50% teaching and 50% research d. Graduate faculty status: Member, Graduate Faculty, 8/2007 — Present e. Education: 2005 Ph.D. Pharmaceutics and Biopharmaceutics, Advisor: Dr. Helen Burt. Thesis: 1998 1994 Effect of Amphiphilic Diblock Copolymers on P-glycoprotein Substrate Permeability in Caco-2 Cells. Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, BC. Canada. MSc. Pharmaceutical Sciences, Advisor: Dr. Keith Simons. Thesis: Evaluation of calcium alginate beads as a prolonged release delivery system for an orally active iron chelator. Faculty of Pharmacy, University of Manitoba, Winnipeg, MB. Canada. B.Sc. Pharmacy, Faculty of Pharmacy, University of Manitoba, Winnipeg, MB. Canada. f. Academic Employment and Positions: 2014 — Present 2007-2014 Associate Professor, Department of Pharmaceutical and Biomedical Sciences, R.C. Wilson College of Pharmacy, University of Georgia, Athens, Ga. Assistant Professor, Department of Pharmaceutical and Biomedical Sciences, R.C. Wilson College of Pharmacy, University of Georgia, Athens, Ga, Associate Member, Interdisciplinary Toxicology Program, University of Georgia, Athens, GA2008 — Present Associate Member, University of Georgia Cancer Center. University of Georgia, Athens, GA. 2008 — Present Associate Member, Center for Drug Discovery. University of Georgia, Athens, GA g- Postdoctoral Positions: 2005 - 2007 Postdoctoral Fellow, Advisor: Dr. Reina Bendayan, Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, University of Toronto. Toronto, ON Canada. 2004-2005 Postdoctoral Fellow, Advisor: Dr. Marcel Bally, BC Cancer Research Center, Advanced Therapeutics. Vancouver, BC Canada. h. Professional Memberships: 2010 — Present Member, American Association for Cancer Research (AACR) 1997 — Present Member, American Association of Pharmaceutical Scientists (AAPS) 2, Teaching: a. Brief description of courses taught at UGA: Pharm. D. Program: 1 PHRM 4200/6200 - Pharmaceutics I: Physical Pharmacy and Dosage Form Design This course was originally taught in tandem, where I was responsible for 50% of the material (Fall 2008 - 2011). I am now the course coordinator and responsible for 100% of the material (Fall 2012). The objectives of this course are to provide students with the basic knowledge of physical pharmacy, pharmaceutics and biopharmaceutical concepts and principles as they apply to the development and assessment of drug delivery systems and dosage forms for the practice of pharmacy. To this end students are exposed to the essentials ‘of thermodynamics, states of matter, and physicochemical properties of drugs and excipients as they relate to the pharmaceutical sciences. Students are expected to comprehend the theoretical and practical pharmaceutical principles necessary for the formulation, production, and quality control of pharmaceutical dosage forms. Ultimately the intent is for students to gain the scientific skills for the evaluation of drug delivery systems and to be able to exercise critical judgment and offer specific recommendations to physicians and patients for optimizing drug therapy. PHRM 4211/6211 - Pharmaceutics I: Principles of Biopharmaceutics and Pharmacokinetics The objectives of this course are to provide students with fundamental knowledge of pharmacokinetic principles including ADME, bioavailability, bioequivalence, therapeuticmonitoring, and interpreting pharmacokinetic parameters as they relate to the practice of pharmacy. My material covers the physiochemical, biological, and drug delivery system factors that impact oral drug bioavailability. Graduate Program: 1. PHRM 8020 - Principles of Pharmaceutical and Biomedical Sciences The objectives of this course are to provide graduate students with a broad range of knowledge to various topics in pharmaceutical and biomedical sciences and research. This includes aspects associated with medicinal chemistry, pharmacology, biopharmaceutics, and pharmacokinetics. I provide lectures on the fundamentals of drug transport across biological membranes and an introduction to drug transporters and pharmacogenomics. 2. PHRM 8270 - Contemporary Concepts in Pharmacokinetics The objectives of this course are to provide graduate students with advanced pharmacokinetic analysis, mathematical modeling, and contemporary issues in pharmacokinetics and biopharmaceutics. My material covered advanced topics on drug transport and bioavailability including in vitro model systems. 3. PHRM 8600 ~ Signal Transduction The objectives of this course are for students to leam the cellular mechanisms of signal transduction by multiple pathways, with specific emphasis on sites for therapeutic intervention and manipulation. For each signaling pathway, the current knowledge of the function, regulation, and structure of the components of the pathway are discussed, and the relevance of the pathway to physiology and pathology will be explored. Prior to a course redesign by the course coordinator, my lectures covered the role of ligand dependent and independent nuclear receptors on gene expression. 4. PHRM 9000 and 9300 — Doctoral Research and Dissertation ‘These graduate courses represent the research and dissertation preparation for completion of the PhD. degree. Students conduct research, participate in weekly lab organized journal club, and present research updates at lab meetings. Students are also expected to present abstracts/posters, at local, national, and intemational meetings, contribute to the writing of manuscripts, and present in departmental seminar. 5. PHRM 8080— Grantsmanship This course covers the fundamentals of developing research ideas, writing grants and their review. My role was to assist the course coordinator in review and provide feedback on student written grant applications. Non-Pharmacy Courses: HONS 4960H, 4970H, 4980H BIOL 4960, 4970, 4980 BIOL 4960H, 4980H, 4980H CBIO 4980 BHSI 4970H MIBO 4960H, 4970HFDNS 4960H, 4990H The objectives of these courses are to provide undergraduate students with an opportunity to participate in pharmaceutical and biomedical research, Students are expected to participate in a lab activities including journal club and provide research updates. Students are evaluated through a written term paper in journal format and an oral presentation. 3, Scholarly Activities: a. Journal Articles: (in print or accepted) — all peer-reviewed 1. C, Mazur, S, Marchitti, and J. Zastre. P-glycoprotein inhibition by the agricultural pesticide propiconazole and its hydroxylated metabolites: Implications for pesticide-drug interactions. Toxicol Lett, 232(1):37-45 (2014). 2. B. Hanberry, R. Berger, and J. Zastre. High Dose Vitamin B1 Reduces Proliferation in Cancer Cell Lines Analogous to Dichloroacetate. Cancer Chemotherapy and Pharmacology. 73(3):585- 594 (2014). 3. R. Sweet and J. Zastre. HIF-1a mediated gene expression induced by Vitamin BI deficiency. International Journal of Vitamin and Nutrition Research, 83(3):188-197 (2013). 4, J. Zastre, C. Dowd, J. Bruckner, and A. Popovici. Intestinal Transport of Pyrethroid Insecticides Using Caco-2 cells. Toxicological Sciences. 136(2):284-293 (2013). 5. J. Zastre, R. Sweet, B. Hanberry, and S. Ye. Linking Vitamin B1 with Cancer Cell Metabolism. Cancer and Metabolism. 1:16 (2013). 6. J. Zastre, B. Hanberry, R. Sweet, C. McGinnis, K. Venuti, M. Bartlett, and R. Govindarajan. Up-Regulation of Vitamin B1 Homeostasis Genes in Breast Cancer. Journal of Nutritional Biochemistry. 24(9):1616-1624 (2013). 7. 8. Hung, H. Mody, S. Marrache, Y. Bhutia, F. Davis, J, Cho, J. Zastre, S. Dhar, C. Chu, R. Govindarajan, Pharmacological Reversal of Histone Methylation Presensitizes Pancreatic Cancer Cells to Anticancer Nucleoside Analogs: Potential Clinical Implications Plos One. DOI: 10.1371/journal.pone.0071196 (2013). 8. O. Kis, J. Zastre, Md. Tozammel Hoque, S. Walmsley, and R. Bendayan. Role of Drug Efflux and Uptake Transporters in Atazanavir Intestinal Permeability and Drug-Drug Interactions. Pharmaceutical Research. 30(4):1050-64 (2013). 9. R. Sweet, A. Paul, and J. Zastre. Hypoxia induced up-regulation and function of the thiamine transporter, SLCI9A3 in breast cancer cells. Cancer Biology and Therapy. 10(11):1101-11 (2010). 10, O. Kis, J. Zastre, M. Ramaswamy, R. Bendayan. pH dependence of OATP2B1 Transporter in Caco-2 cells: Potential Role in Antiretroviral Drug Oral Bioavailability and Drug-Drug Interactions. Journal of Pharmacology and Experimental Therapeutics. 334(3):1009-1022 (2010).11. J. Zastre, G. Chan, P. Ronaldson, M. Ramaswamy, P. Couraud, I. Romero, B. Weksler, M. Bendayan and R. Bendayan. Up-regulation of P-glycoprotein by HIV Protease Inhibitors in a Human Brain Microvessel Endothelial Cell Line. Journal of Neuroscience Research 81(4):1023- 1036 (2009). 12. J. Zastre, J. Jackson, W. Wong, and H. Burt. P-glycoprotein Efflux Inhibition by Amphiphilic Diblock ‘Copolymers: Relationship between copolymer concentration and substrate hydrophobicity. Molecular Pharmaceutics 5(4):643-653 (2008). 13.N. Chattopadhyay, J. Zastre, H. Wong, S. Wu and R. Bendayan. Nanoparticles enhance the delivery of the HIV protease inhibitor, atazanavir, by a human brain endothelial cell line. Pharmaceutical Research 25(10): 2262-2271 (2008). 14, E, Ramsay, M. Anantha, J. Zastre, J. Zonderhuis, M. Meijs, D. Strutt, M. Webb, D. Waterhouse, and M. Bally. Irinophore C: A liposome formulation of irinotecan with substantially improved therapeutic efficacy against a panel of human xenograft tumors. Clinical Cancer Research 14(4):1208-1217 (2008). 15. E, Ramsay, J. Alnajim, M. Anantha, J. Zastre, H. Yan, M. Webb, D. Waterhouse, M. Bally. A novel liposome irinotecan formulation with significant anti-tumour activity: Use of the divalent cation ionophore A23187 and copper-containing liposomes to improve drug retention. European Journal of Pharmaceutics and Biopharmaceutics 68(3):607-617 (2008). 16.5. Zastre, J. Jackson, W. Wong, and H. Burt. Methoxypolyethylene glycol-block- polycaprolactone diblock copolymers reduce P-glycoprotein efflux in the absence of a membrane fluidization effect while stimulating P-glycoprotein ATPase activity. Journal of Pharmaceutical Sciences. 96:864-875 (2007). 17. J. Zastre, E. Ramsay, M. Anantha, and M, Bally. Irinotecan-cisplatin interactions assessed in cell based screening assays: cytotoxicity, drug accumulation and DNA adduct formation in an NSCLC cell line. Cancer Chemotherapy and Pharmacology. 60:91-102 (2007). 18, D, Waterhouse, K. Gelmon, R. Klasa, K. Chi, D. Huntsman, E. Ramsay, E, Wasan, L. Edwards, C. Tucker, J. Zastre, Y. Zhang, D. Yapp, V. Dragowska, S. Dunn, S. Dedhar, and M. Bally. Development and assessment of conventional and targeted drug combinations for use in the ‘treatment of aggressive breast cancers. Current Cancer Drug Targets 6:455-489 (2006). 19. J. Zastre, J. Jackson, and H. Burt. Evidence for modulation of P-glycoprotein mediated efflux by methoxypolyethylene glycol-block-polycaprolactone amphiphilic diblock copolymers. Pharmaceutical Research 21:1489-1497 (2004). 20. K. Letchford, J. Zastre, R. Liggins, and H. Burt. Synthesis and characterization of short block length methoxypoly(ethylene glycol)-block-poly(caprolactone) diblock copolymers as micellar drug delivery systems. Colloids and Surfaces B: Biointerfaces 35:81-91 (2004). 21.J. Zastre, J. Jackson, M. Bajwa, R. Liggins, F. Iqbal, and H. Burt. Enhanced cellular accumulation of a P-glycoprotein substrate, rhodamine-123, by caco-2 cells using low molecular weight methoxypolyethylene glycol-block-polycaprolactone diblock copolymers. European Journal of Pharmaceutics and Biopharmaceutics 54:299-309 (2002). 22.N. Bamabe, J. Zastre, S. Venkataram, and B. Hasinoff. Deferiprone protects against doxorubicin-induced myocyte cytotoxicity. Free Radical Biology & Medicine 33:266-275 (2002).b, Journal Articles: (Under peer-review or revision) . Book Chapters: Peer reviewed 1, J. Bruckner, T. Osmitiz, S. Anand, D. Minnema, W. Schmitt, N. Assaf, and J. Zastre. Chapter 5: The Influence of Maturation on Rat and Human Physiological Processes Involving Protein and Lipoprotein Binding, Gastrointestinal Absorption, Blood Brain Permeability and Transport of Pyrethroids. ACS Symposium Series 1099 Parameters for Pesticide QSAR and PBPK/PD Models for Human Risk Assessment. Editors: J. Knaak, C. Timchalk, and R. Tomnero-Velez (2012). d. Abstracts: (selected out of 33) 1, J. Zastre, B. Hanberry, B. Sweet, K. Venuti, C. McGinnis, M. Bartlett, R. Govindarajan. Up- regulation of Vitamin B1 Homeostasis Genes in Breast Cancer. AACR Annual Conference. ‘Washington DC, April 2013 2. B, Sweet, L. Lim, J. Zastre. Thiamine deficiency induces HIF-1a stabilization and target gene expression. AACR Annual Conference. Washington DC, April 2013. J. Zastre, C. Dowd, K. Venuti, D. Gullick, M. Bartlett, D. Minnema, D. Gammon, S. Anand, J. Bruckner. Absorption & Transport of the Pyrethroid Insecticide Deltamethrin (DLM) by Caco-2 Cells. FASEB, Boston MA 2013 4, R. Sweet, R. Govindarajan, and J. Zastre. Adaptive Post-translational Regulation of the Thiamine Transporter, THTRI in Breast Cancer cells During Hypoxic stress. AACR annual meeting, Chicago March 2012. 5, R, Sweet, R. Govindarajan, and J. Zastre. Hypoxia-Mediated Intracellular Redistribution of the Thiamine Transporter THTR1 towards the Plasma Membrane. AAPS Annual Conference, Washington, DC. (Oct 2011). 6. B. Hanberry and J. Zastre. The role of TPK1 in breast cancer cell proliferation during hypoxic stress. AAPS Annual Conference, Washington, DC. (October 2011). 7. R. Sweet, R. Govindarajan, and J. Zastre. Hypoxia-Mediated Intracellular Redistribution of the Thiamine Transporter THTR1 towards the Plasma Membrane. UGA GSPS Research Day, Athens, GA (May 2011). 8, R, Sweet, A. Paul, R. Govindarajan and J. Zastre. Alterations in the Expression and Intracellular Localization of the Thiamine Transporters SLC9A2 and SLCI9A3 during Hypoxic Stress. ‘AAPS Workshop: Drug Transporters in ADME, Bethesda, MD (March 2011). R. Sweet, A. Paul, and J. Zastre. Hypoxia induced up-regulation of the thiamine transporter, SLC19A3, in breast cancer cell lines. AAPS Annual Conference, New Orleans, LA (November 2010). 10. O. Kis, J. Zastre, M. Ramaswamy, and R. Bendayan, Role of Intestinal membrane transporters in antiretroviral drug absorption and drug-drug interactions. AAPS Annual Conference, New Orleans, LA (November 2010). ©11.L, Willis, A. El-Remessy, J. Zastre, S. Fagan. Angiogenic Capacity of Immortalized Human Microvascular Endothelial (xCMEM/D3) Cells Following Oxygen-Glucose Deprivation. MCG Graduate Research Day (March 2010). 12, R. Sweet, A. Paul, and J. Zastre. Hypoxia induced up-regulation of the thiamine transporter, SLC19A3, in breast cancer cell lines. UGA GSPS Research Day, Athens, GA (2010). 13.F. Kahn, and J. Zastre. Examining the Substrate and Inhibitor Properties between Atazanavir and Pravastatin with the OATP2B1 Transporter. UGA CURO Symposium (2010). 14. A. Paul and J. Zastre. Hypoxia Induced Up-Regulation of the Thiamine Transporter SLC19A3 in Breast Cancer Cell lines. Georgia Cancer Coalition Annual Meeting (2009). 15.8. Jun, A. Paul, and J. Zastre. The Famesoid-X Receptor and Associated Target Genes are up- regulated under Hypoxia in Breast Cancer Cell Lines. Georgia Cancer Coalition Annual Meeting (2009). 16. A. Ibrahim, J. Zastre, and R. Amold. Impact of tumor hypoxia and dosing schedule of paclitaxel and topotecan in human breast and prostate cancer cell lines in vitro. Georgia Cancer Coalition Annual Meeting (2009). 17.D. Li, C. White, and J. Zastre. Induction of Pregnane X receptor by Atazanavir and Consequential Effects on Tissue Distribution of Atazanavir in Mice. AAPS Annual Conference, Los Angeles CA (2009). 18. O. Kis, J. Zastre, A. Otting, M. Ramaswamy, and R. Bendayan. Interactions of HIV-1 protease inhibitors with the OATP2B1 influx transporter in Caco-2 cells. APS Annual Conference, Atlanta Georgia (2008). 19.G. Chan, J. Zastre, M. Ramaswamy, M. Bendayan, and R. Bendayan. Upregulation of the Efflux Drug Transporter, P-glycoprotein (P-gp), by HIV Protease Inhibitors. AAPS Annual Conference, Atlanta Georgia (2008). e. Invited Oral Presentations 1. Can a Vitamin Promote Malignant Progression? 2014 Making Strides Kickoff, American Cancer Society. Atlanta, Georgia. August 28" 2014 2. The Duality of Vitamin B1. American Cancer Society. Athens, GA. June 132014 3. Impact of Vitamin B1 on Malignant Progression. Department of Pharmacal Sciences, Harrison School of Pharmacy, Auburn University. Aubum, AL. Feb 12, 2013 4, Impact of Vitamin B1 on Malignant Progression. Department of Biochemistry & Molecular Biology, Cancer Research Center, Georgia Health Sciences University. Augusta, GA. Jan 18, 2013, 5. Can Vitamin B1 Promote Malignant Progression? University of Manitoba, Faculty of Pharmacy. Winnipeg, MB Canada Nov 7, 20126. Targeting Micronutrient Transporters within Hypoxic Tumor Microenvironments. Georgia State University, Department of Chemistry. Dec 2010. 7. Exploiting Tumor Heterogeniety: Targeting Micronutrient Transporters within Hypoxic Tumor Microenvironments. UGA Cancer Center Retreat Jan 2010. 8 Exploiting Tumor Heterogeniety: Improving Anticancer Drug Targeting to Hypoxic Tumor Microenvironment Phenotypes. UGA Center for Drug Discovery Oct 2009. 9. Exploiting Tumor Heterogeniety: Improving Anticancer Drug Targeting to Hypoxic Tumor Microenvironment Phenotypes. UGA Computational Systems Biology Seminar. July 2009. f. Research Grants: Current Funding: 1. NIH 1 R21 AA021948-0 06/02/13 — 05/31/15 Role - PI (Zastre/Bartlett) $353,875 (Total) Title: Adaptive Regulation of Vitamin BI Transport The overall objective we have for this application is to determine the transcriptional pathways involved in the adaptive regulation of thiamine transporters during thiamine deficiency associated with chronic alcoholism. 2. American Cancer Society 07/01/14 — 06/30/18 Role — PI (Zastre/Bartlett/Wang) $ 792,000 (Total) Title: Role of Vitamin BI in Hypoxia Mediated Malignant Progression ‘The overall goal for this application is to determine the functional importance of thiamine homeostasis within hypoxic cancer microenvironments. Pending Grant Applications Completed: 1. Council for the Advancement of Pyrethroid Human Risk Assessment (CAPHRA) Role - Co-Pl (Bruckner/Bartlett/White/Cummings/Zastre) 1/01/11 - 10/31/14 $960,164 (Direct) Title: Characterization of Age-Related Differences in the Pharmacokinetics of Pyrethroids The goals of this project are to determine age-dependent effects of specific physiological processes in the pharmacokinetics of pyrethroid insecticides. In particular, my contribution is to assess the cellular mechanisms facilitating oral absorption and blood brain barrier permeability 2. Georgia Cancer Coalition (GCC) Role — PI (Zastre) 07/01/08 - 06/30/13 GCC Distinguished Cancer Scholar $250,000 (Direct) Title: Role of Solute Carrier Transporters in Cancer The overall goal for this project is to understand the functional role of solute carrier transporters as mediators of malignant progression and chemosensitivity in cancer.3. Faculty Endowment Seed Grant - ($2,000) Awarded to Dr. Zastre. Title: Defining Dietary Influences and Chemotherapeutic Strategies towards Malignant Progression 4. Council for the Advancement of Pyrethroid Human Risk Assessment — ($84,000) Awarded to Drs. Bruckner, White, Bartlett, and Zastre. Feasibility study on the Characterization of Age- Related Differences in the Pharmacokinetics of Pyrethroids 5. Faculty Endowment Seed Grant — (84,000) Awarded to Dr. Zastre entitled In Vivo Determination of Hypoxia within a Spontaneous Breast Cancer and Tumor Xenograft Mouse Model. 6. Faculty Endowment Seed Grant - ($6,000) Awarded to Drs. Zastre and Catherine White. Title: Role of Pregnane-X Receptor in Distribution of Protease Inhibitors to HIV Reservoirs: Brain and Testes. 7. Interdisciplinary Toxicology Program Small Equipment Grant — ($3,500) Awarded to Drs. Amold, Zastre, Cummings, Bruckner. The matching funds were used to upgrade a Nikon AZ- 100 multipurpose fluorescent stereomicroscope with a new digital color camera, mounting bracket, switcher and luciferase filter and cube. 8. Faculty Endowment Seed Grant - ($4,000) Awarded to Drs. Jason Zastre and Catherine White entitled “Role of Pregnane-X Nuclear Receptor in limiting Maternal to Fetal Drug Transport”. 9. Faculty Endowment Seed Grant - ($4,000) Awarded to Drs. Jason Zastre and Robert Amold entitled “Targeting Hypoxic Mediated Alterations in Transporters with Drug Nanoparticles”. 10. Faculty Endowment Seed Grant - ($1,200) Awarded to Drs. Jason Zastre and Robert Amold entitled “Microarray profiling of gene expression alterations in hypoxic human breast and prostate cancers”, g. Honors 2014 American Cancer Society Research Scholar 2008 Georgia Cancer Coalition Distinguished Scholar (AKA: Georgia Research Alliance Distinguished Cancer Scientist) 2005-2007 CIHR-Rx&D Postdoctoral Fellowship 2000-2003 Science Council of British Columbia GREAT Scholarship 1999 University of British Columbia Graduate Fellowship 1995 Manitoba Health Research Council Fellowship 1994 Apotex P.A.C.E. Future Leader Award 1994 Merck Frosst Graduate Scholarship 1993 Glaxo Ltd. Scholarship 1993 Medical Research Council Summer Undergraduate Fellowship 1993 Novopharm Ltd. Scholarship 1993 Flexon Silver Medalh, Outside press and comments related to my research Below are listings of web articles from local, national, and international web news services Chypetinks) and reference of a published comment regarding my peer reviewed manuscript. http://onlineathens.com/features/2011-10-29/u lh 1011-01 -vitamin-therapy-breast-cancer-patients. html //onlineathens, com/uga/2012-03-30/toxicologis insecticide-risks-infi ‘http://www.highbeam,com/doc/1G1-246966262.htm! //www.healthjockey.com/2011/01/21/vitamin-t ( -for-| 7 perjudicial-para-los-pacientes-con-cancer-de-mama Richardson AD, and Moscow JA: Can an enzyme cofactor be a factor in malignant progression? Comment on: Hypoxia induced upregulation and function of the thiamine transporter, SLC19A3 ina breast cancer cell line. Cancer Biology and Therapy 10:1112-1114 (2010). 4. Service Contributions: Professional: Invited reviewer Apoptosis Molecular Pharmaceutics Pediatrics Critical Reviews in Oncology/Hematology Current Pharmaceutical Design Journal of Pharmacology and Experimental Therapeutics Journal of Controlled Release Toxicological Sciences Drug Metabolism and Disposition European Journal of Pharmaceutics and Biopharmaceutics Department: ITP Graduate Admissions Committee member 2012 — Present PBS Graduate Committee member 2014 — Present Graduate Education Task Force member 2011 - 2012 Graduate Admissions Committee member 2008 - 2011 College: Pharm D Admissions Committee member 2012 — Present Curriculum Committee member 2014 — PresentCurriculum Committee member Graduate Education and Curriculum Committee chair Graduate Education and Curriculum Committee member University: Lead for DDD interdisciplinary Group — ILS Judge - Graduate Student Research Day Community/outreach: High school science fair research mentor 2009 - 2012 2009-2011 2008 - 2009 2014 - Present May 2010 and May 2011 2011EXHIBIT BFiticiinic Labs Scientific Report XRPD, DSC, and IR Analysis of Pentobarbital Samples Georgia Department of Corrections Prepared for Mr. Robert Jones April 2, 2015 Project Number 2015051 Report Number R201583.01 Author: Bact 0. zien __Apci] 2,200. Brett D. Bobzien, Sr. Scientist Date Reviewer: < Veli David E. Bugay, Ph.D ESO DateSummary One pentobarbital powder sample and a syringe containing a solution with suspended solids provided by Georgia Department of Corrections were analyzed by x-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and infrared (IR) spectroscopy. Information for the samples provided is listed in Table 1 Table 1. API Samples Provided ; Tinie Sample ‘Sample Description nie Serr Pentobarbital Sodium, CAS# 57-33-0, Purity 99.00%, Case # - |I-2015-1214 ed Pentobarbital Sodium, SOmg/ml, Exp, 03/24/2015, Case #--2015-1214 | T2759 Results The filenames for the data collected from the powder sample and the solids from the syringe are summarized in Table 2. The powder sample was able to be analyzed as received. The solids from the syringe were isolated by centrifuging the material, decanting the liquid, and allowing the material to air dry. Table 2. Analytical Testing Filenames Tricinic Sample XRPD | DSC |DSCPage] IR | IRPage Number __| XRPD Filename | page No. | Filename | No. | Filename | No. TOL2758. PX-7453 6 | osc20s1t| 9 1R782 it TL2759 RXI-7456 6 | sc2.0812 | 10 R783 it ‘An overlay of the XRPD patterns for the powder sample and the solids from the syringe is displayed in Figure 1. The pattern for the powder sample provided shows a number of broad discrete diffraction peaks overlaid on a raised baseline, suggesting that this, sample contains crystalline and non-crystalline material. The pattem for the solids isolated from the syringe showed a number of broad, discrete, diffraction peaks superimposed on a flat baseline, indicating that this sample is crystalline. Visual comparison of the patterns showed that there were some similar peaks between the patterns, but a number of unique peaks were also observed in the pattems. In order to determine the identity of the materials in the samples, a database searchimatch analysis was performed. Figure 2 shows an overlay of the XRPD pattern for the powder sample with the database stick patterns for one solid form of pentobarbital sodium, one solid form of pentobarbital, and sodium chloride [1]. These three stick pattems describe all but two broad, low-intensity, high-angle reflections, observed in the XRPD pattem for the powder sample. This would suggest that these three components comprise the majority of the sample. Figure 3 shows an overlay of the XRPD pattem for the solids isolated from the syringe and the database stick patterns for two different solid forms of pentobarbital [2]. These two stick patterns R201583.01 Page 2 of 13describe all but four low-intensity high-angle reflections observed in the pattem for the solids isolated from the syringe. This would suggest that these two components comprise the majority of this sample. The results from the database search for the two patterns would suggest that the two samples contain one common form of pentobarbital, but that the other components comprising these samples are different. The powder sample appears to contain pentobarbital sodium as its other major component, while the solids isolated from the syringe contain another polymorph of pentobarbital as the other major component. This ‘would indicate that the sodium salt of pentobarbital broke apart when preparing the solution in the syringe and polymorphs of the free base ultimately precipitated from the solution in this sample. ‘The powder sample and the solids isolated from the syringe were also analyzed by DSC and the thermograms are presented in Figures 4 and 5, respectively. The DSC results are summarized in Table 3. Table 3. DSC Results Triinic Sample | DSC . Number Filename DEC Rests Toi2768 | DSC2.0917 | Endos. at ~143 °C (), 290 °C (B), 305°C (a), and 316°C (@) Tel2759 | DSC2.0912 Endos. at ~76 °C (b), 140 °C (b), and 246 °C (b) 'a. Endos. = endotherms; b= broad; s = sharp. The thermograms for the two samples are quite different, with the only similarity being that there is an event between 140 and 145 °C for both samples. This would suggest that the compositions of the two samples are largely different, but that there may be a common component. This is consistent with the XRPD findings which suggested that there was a common component between the samples, but other components were different. An overlay of the IR spectra for the two samples is presented in Figure 5. As with the XRPD and DSC data, there are similarities between the spectra for the two samples. A number of common bands are observed between the two spectra, but there are also regions that show significant differences. A library search was performed for the powder sample and a solid match was found to a library spectrum of pentobarbital sodium [3]. The spectrum for the solids isolated from the syringe contained a number of additional bands that were not observed in the library spectrum of the pentobarbital sodium. No suitable matches were found that described these additional spectral bands, R201583.01 Page 3 of 13Conclusions Based on the analyses performed in this study, the powder sample is believed to contain pentobarbital sodium, a polymorph of pentobarbital free base, and sodium chloride. The solids isolated from the syringe are believed to contain two different polymorphs of pentobarbital free base. Experimental XRPD The Rigaku Smart-Lab X-ray diffraction system was configured for reflection Bragg- Brentano geometry using a line source X-ray beam. The x-ray source is a Cu Long Fine Focus tube that was operated at 40 kV and 44 mA. That source provides an incident beam profile at the sample that changes from a narrow line at high angles to a broad rectangle at low angles. Beam conditioning slits are used on the line X-ray source to ensure that the maximum beam size is less than 10 mm both along the line and normal to the line. The Bragg-Brentano geometry is a para-focusing geometry controlled by passive divergence and receiving siits with the sample itself acting as the focusing component for the optics, The inherent resolution of Bragg-Brentano geometry is governed in part by the diffractometer radius and the width of the receiving slit used. Typically, the Rigaku Smart-Lab is operated to give peak widths of 0.1 20 or less. The axial divergence of the X-ray beam is controlled by 5.0-degree Soller slits in both the incident and diffracted beam paths. ‘The powder samples were prepared in a low background Si holder using light manual pressure to keep the sample surfaces fiat and level with the reference surface of the ‘sample holder. The single crystal Si low background holder has a small circular recess (7 mm diameter and about 1 mm depth) that holds between 15 and 25 mg of powdered material. Each sample was analyzed from 2 to 40 °28 using a continuous scan of 6 °26 per minute with an effective step size of 0.02 °28. Dsc DSC analyses were performed using a TA Instruments Q2000 differential scanning calorimeter equipped with a refrigerated cooling system (RCS). Approximately 3 to 4 mg of each sample was weighted into an aluminum Tzero pan, covered with and aluminum Tzero lid, and crimped. The sample cell was heated from ambient to 350 °C ata rate of 10 °C/minute. An empty reference pan prepared in the same way was also present in the cell to account for the heat flow properties of the pan. The DSC instrument is controlled using the Thermal Advantage Release 5.2.5. infrared Spectroscopy — IR ‘The IR spectra were acquired utilizing a Thermo Nicolet model 6700 Fourier-transform (FT)R spectrophotometer equipped with a deuterated triglycine sulfate (OTGS) detector, a potassium bromide (KBr) beamsplitter, and an electronically temperature controlled (ETC) Ever-Glo® IR source. The spectra were acquired using a SMART iTR diamond attenuated total reflectance (ATR) sampling accessory with a spectral range of R201583.01 Page 4 of 134000-525 cm". Each spectrum is the result of 128 co-added scans acquired at 2 cm’ resolution. A single beam background scan of the air was acquired before the sample scan allowing presentation of the spectrum in Log 1/R units. A wavelength calibration ‘was performed using polystyrene. (The Omnic 8.2 software package (Thermo-Nicolet) was used to acquire, process, and evaluate the spectral data) References 1. International Centre for Diffraction Data PDF-4 2015 Organics Database, downloaded and interfaced with Rigaku PDXL software at Triclinic Labs, card numbers 00-029-1920 and 00-027-1597. 2. International Centre for Diffraction Data PDF-4 2015 Organics Database, downloaded and interfaced with Rigaku PDXL software at Triclinic Labs, card numbers 00-028-1648, 3. Sigma Biological Sample Library, Sample Index #9, Copyright 2008 Thermo Fisher Scientific Inc. R201583.01 Page 5 of 13e1y09e6ed voreestozu weuzeouer oa ceuzeeureer oat — ‘SBUUAS Oty Woy PayEI0S! Spljos Uy pue ejdules sepmod jeyiqueqoyued eu Jo) sWeHed GEYX Jo AEUEAO “| eunBi4Figure 2, Overiay of the XRPD pattem for the powder sample with the database patterns for pentobartbital sodium. Qualitative Analysis Results nay ote zoisouca 118722 Seer Maruomertdos ——_—Z018027 084038 Beene x17485_Thelo_2- Spar Rate commer Fiasenee__—_—— Fam Egueeined Praca aad De a Selon She — Gr FATES — Tet (eOS OFS coaaetazt Seige TTS 18 16D BORA ——eoga7-887 ra Cina Base ear (Coos) Feast PEE 4 eee t = 8cet008- 7 Ea t cra a 6.0e+003- 3 ee E 3 g . aE > =e i 2 af a é Se oe B acoso 3s i z 3 ; 24 2.00+003 tee | 40 2.theta (deg) 201583.01 Page 7 of 13Figure 3. Overlay of the XRPD pattern for the solids isolated from the syringe with the database patterns for two polymorphs of pentobarbital Qualitative Analysis Results Analysis date aotsiosoa 12.1447 pe name Measurement date 201803727 08:46:96, Fie name ax1-7454_Theta_2- Operator Theta TXT ‘comment Qualitative analysis results Phase rane Foul Figure ofmert Phase req deta OB card number Sethe cr HieN203 1.255, ICDD (POF 00-07-1597, Satyr CriHieN203_—1.468 ICDD [POF-¢ 00:028-1648. EEE z Sry ss Tee 1.s0+004 E SSE z 5 E TE Hsrec ce nm = $2 Heike af fo Pe ety; ipa a iB BZEE: 2 : } qLEE: 8 3 3 BPs 3h = 1.06+004+ : PEITE = 3 z fe: a¢ : 2 2 aS243 2 5 q ge2ag 5 = £ fii gs 2 | z BE: a} z 5.0e+003, 2 gis 7 : Rice = ‘ £2 F & Ei EE 3 Bf ee ‘ 2.thata (deg) 201583.01 Page 8 of 13Figure 4. DSC thermogram for pentobarbital powder sample. ‘Sample: TCL2787 Fie: T:..2015051\DatalDSC\DSC2.0911 ‘Size: 2.4250 mg DSc Operator: TLC Method: Ramp Run Date: 26-Mar-2015 15:58 Instrument: DSC Q2000 V24.11 Build 124 ° z é i site £ 4 3 % wo Bo 20 20 aio to wou Temperature (°C) R201583.01 Page 9 of 13Figure 5. DSC thermogram for soli isolated from the syringe. Universal V4 5A TA Instruments sample: 204902 Fe: T.120180511DatadDSC\08C2.0912 Size: 5.6540 mg DSC Operator: TLE Nei Ramo Ron bate: Pear 2018 168 Return D&C C2000 W241 Bull 124 os 0. B os = 3 nie B a0 nesec fae 139.58°C_ 20 2 10 io 20 20 ato 60 B0 Up ‘Temperature (°C) R201583.01 Page 10 of 13#140 14 bed boreestozu ‘eBUUAS @Uy Woy PeyE|0S! Spljos ely pue ejdwles sopmod jen1queqo}ued 104 ExDads yl OU) Jo AEWEAO “9 @:N6L4Figure 7. Overlay of the IR spectrum of the pentobarbital powder sample and a library spectrum for pentobarbital sodium. Search osu for. TCL2TST: Pentabarttl Data: Thu Apr 02 16:15:48 2015 (GMT-04-00) ‘Search ago: Search Export Regions searched 450,002800.00 (CLBTBT, Peniobarbital [a € ¥ oa oo 1S PENTORARETATSODIUN-CEN Son Match83.44 Abvorbues g ‘9500 ‘3000 72500 ‘2000 1500 1900 Wavenumbers (cm-1) ‘Search experts comments ‘Soarehrsuts st of matehos Index Match Compouna Namo rary Name 19 3.44 PENTOBARBITAL SODIUM-DEA SCHEDUL ‘Signa Biologia! 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